PMID- 11725600 TI - Can we use the enablement model to end the feud? PMID- 11725601 TI - Laboratory methods for allergen extract analysis and quality control. PMID- 11725602 TI - Regulatory aspects of allergen vaccines in the US. PMID- 11725604 TI - Molecular characterization of allergens. PMID- 11725603 TI - Cross-reactivity of plant and animal allergens. PMID- 11725606 TI - Skin testing and immunoassays for allergen-specific IgE. PMID- 11725605 TI - Recombinant allergens. PMID- 11725607 TI - Immunotherapy for allergic diseases. PMID- 11725608 TI - Common allergenic pollens, fungi, animals, and arthropods. PMID- 11725609 TI - Allergens. PMID- 11725610 TI - The manufacture of allergenic extracts in North America. PMID- 11725611 TI - [Strategy for international health cooperation: the example of the Monkole Hospital Center of Kinshasa]. PMID- 11725612 TI - [Genetic therapy in oncology: ethical aspects]. AB - The more advanced oncologic therapies are directing toward new frontiers, on account of the remarkable undesirable effects of chemio- and radio-therapies. This new therapeutic experiences are of type biological (vaccines), or genic (substitution again genes with shutters meaning-tumoral). This therapies involve, to be effected, some ethical shrewdnesses: choice of the patient, the engineering modality of the genes, the transfer of the genes in cells of the exclusively somatic line, the elimination of the pathogenic risk of the vector virus, the obligatory use of sterile rooms, the attention to the administration of the drug, a legal issue of the judgment of notoriety. PMID- 11725613 TI - [Identification of "presumptive risk" of hypertension crises with fractal interpolation of the 24-hour arterial pressure. III: study of normotensive subjects]. AB - PURPOSE: The present study is devoted to identify the normotensive subjects who are at a "presumptive risk" (PR) for hypertensive crisis, by applying the "fractal interpolation" (FI) to blood pressure 24-h pattern. MATERIALS AND METHODS: The investigation was performed on 109 ascertained cases of normotension, who underwent a non-invasive ambulatory monitoring of their 24-h blood pressure, by applying the FI to their daily mean arterial pressure. RESULTS: The study showed that the PR for hypertensive crisis can be found in 12% out of the investigated normotensives. The proportion of such a risk is not significantly higher in non-dippers as compared to dippers. Vice versa, the prevalence of cases with a PR for hypertensive crisis was found to be significantly increased in normotensives who were deprived by a significant blood pressure circadian rhythm as compared to those who showed the blood pressure circadian rhythm to be preserved. Furthermore, the cases "at risk" were found to show a different spectrum for the harmonic formants which compose the complex harmonic structure of the blood pressure 24-h variability. CONCLUSIONS: The PR for hypertensive crisis in normotensives seems to be increased by the loss of the blood pressure circadian rhythm, due to a change in the harmonic structure which confers the 24-h periodicity to the hemodynamic variable. This means that the abrogation of the blood pressure circadian rhythm in conditions of normotension promotes an adjunctive disorder for which the PR for hypertensive crisis is augmented. PMID- 11725614 TI - [Treatment of euthyroid goiter in the elderly]. AB - PURPOSE: A study was conducted to evaluate the effectiveness of the medical therapy with synthetic hormone levothyroxine (L-T4) in the elderly subjects with multinodular euthyroid goiter. PATIENTS AND METHODS: 187 elderly subjects (34 males and 153 females) ranging between 63 and 85 years of age with multinodular euthyroide goiter were examined. For each subject has been calculated the index of body mass (BMI) which has consented the identify two groups of subjects: the elderly patients with normal weight and the obese subjects. RESULTS: In the mostly of the patients (82%), both normal weight and obese, the L-T4 therapy has not determined significant changes either of the dimensions or the number of the nodules. In the obese subjects the L-T4 therapy has not caused decrease of weight at least to the 20% of the initial body weight. CONCLUSIONS: The results of the research have proved the limited effectiveness of the suppressive therapy with levothyroxine in the reduction of the volume and/or of the number of the nodules, without however denying the usefulness in the preventing the worsening of the nodular disease of thyroid. The study also has revealed that the therapy with levothyroxine is ineffective for the body weight reduction in the obese subjects. PMID- 11725615 TI - [Quality of life in menopause. Experimental research]. AB - Psychophysiological relationship in menopausal women was explored by the comparison of stress level and resistance to stress and related symptoms: socio economic and cultural variables and willingness to participate into a specific therapeutic program were taken into account. One hundred women, aged 44-59 years (54.2 +/- 5.64), that consecutively were referred to an out-patient menopausal Clinic, were studied by a questionnaire that evaluates quality of life (qol) derived and adapted from the Sickness Impact Profile and the Functional Living Test; moreover the test MSP (psychological stress measurement), translated from the original and adapted to Italy, was used. Psychological complaints of women examined are minimal: adaptation tests are substantially normal, and stress index measurements are even lower in comparison with a gender and age matched population group. Our results suggest that after a short period of psychological tension at the onset of climaterium, women acquire a positive, stable status, well different from stereotypes and prejudices around psychological disturbance associated with menopause. Minimal relevance of perceived disturbance seems the reason that limits the women's need and request for a specific therapy. Menopausal depression seems more related to life changes than to hormonal alterations. Reduced physical fitness, increased risk of parental death, difficulty to cope with new roles, apart the care of adult sons or daughters. A critical point is socio-economic level and the possibility to maintain a satisfying work. Contextual conditions, and specially degree of instruction, type of role change inside the family and number of sons, appears the most evident and relevant variables that mediate psychophysical conditions and perceived quality of life. PMID- 11725616 TI - The role of nasogastric tube after elective abdominal surgery. AB - PURPOSE: Nasogastric tube (NGT) is routine employed after abdominal surgery still in our country and abroad too, but the patients' discomfort is severe and several serious complications were referred as related to its use. The aim of this study was to evaluate the pros and cons of routine use of NGT in elective uncomplicated abdominal surgery. PATIENTS AND METHODS: In order to evaluate the routine use of NGT we performed a prospective randomized trial on 100 patients who had elective uncomplicated abdominal surgery: 50 subjects had the early removal of NGT and in 50 patients it was maintained until passage of flatus/feces. RESULTS: The relevant differences between the two groups were the earlier passage of flatus and feces and the lower incidence of postoperative gastrectasy in the group where NGT was early removed. No statistical difference was recorded concerning the occurrence of postoperative nausea, vomiting, abdominal distension, complications and day of clinical release. CONCLUSIONS: The routine application of NGT in elective abdominal surgery could be omitted if the surgeon and nursing team are willing to renounce a useful tool in providing informations about the resolution of postoperative intestinal atony to the patient's benefit: in fact a severe discomfort due to the NGT was recorded in the 70% of our series. PMID- 11725617 TI - [New treatments for arthrosis]. PMID- 11725618 TI - [Nf-kB transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications]. AB - PURPOSE: Description of the involvement of the transcription factor NF-kB in inflammatory, autoimmune and neoplastic processes. Clinical implications from basic research. DESIGN: Review of the most significant data reported in the literature and personal publications. RESULTS: NF-kB is an ubiquitous transcription factor member of the proto-oncogene family rel. NF-kB regulates the expression of several genes involved in inflammatory and immune responses. The classical activated form of NF-kB consists of the p50/p65 heterodimer, different dimers may be formed with members of rel, AP1, steroid hormones receptors family. Many studies suggest that NF-kB should be considered as an important mechanisms of inflammatory processes and autoimmune diseases. Many important anti inflammatory drugs and immunosuppressants inhibit NF-kB. Several observations have suggested a role of the inappropriate activation of NF-kB in cell proliferation, transformation, and tumor development, mainly lymphomas. Conversely, it has been proposed that the activation of NF-kB in immune cells may contribute to anti-tumor immunity. CONCLUSIONS: NF-kB is an optimal target of anti-inflammatory and immunosuppresant therapies. Molecular studies on NF-kB are very important to understand the pathogenesis of inflammatory, autoimmune and neoplastic diseases, and to identify new drugs that inhibit NF-kB activation. PMID- 11725619 TI - [Anaplastic carcinoma of the thyroid: diagnosis and treatment]. AB - Anaplastic thyroid carcinoma (ATC), accounting for 5% to 15% of primary malignant thyroid neoplasm, is one of the most aggressive solid tumors in humans. It is rapidly fatal, with a mean survival of 6 months after diagnosis. Multimodality treatment with surgery and/or external beam radiotherapy and chemotherapy are of fundamental importance for local control of disease and to enhance survival. Molecular biology studies have shown that ATC is associated with a p 53 mutation. ATC usually does not concentrate radioiodine or express thyroglobulin. It is essential to verify the diagnosis histologically because insular thyroid cancer, lymphomas, and medullary thyroid cancer are occasionally confused with undifferentiated neoplasms. Immunohistochemical study is helpful in establishing the diagnosis. Multimodal therapy and development of effective systemic chemotherapy agents would provide to result in improvements in survival although no single agent has yet been identified. Aggressive multimodality treatment regimens show promise in improving local control in patients with ATC. Survival rates however remain low. Despite intense applications of such integrated therapy, no standardized successful treatment protocol has yet been established. PMID- 11725620 TI - Huge response to letrozole in inoperable T4 breast cancer: a case report. AB - A 73 years-old woman presented with locally advanced breast cancer (cT4b cN1 M0, stage IIIB) and atrial flutter. Because of the arrhythmia, chemotherapy or tamoxifen, although malignancy was hormone-sensitive, were discarded. Letrozole was started. Two months later, the breast nodule and skin ulceration cleared up. Surgery was performed. Nowadays, 24 months later, patient does fine, continuing letrozole as adjuvant treatment. PMID- 11725621 TI - [Early clinical applications of radiotherapy]. PMID- 11725622 TI - Otitis media in Greenland. Studies on historical, epidemiological, microbiological, and immunological aspects. AB - This thesis describes the different aspects of otitis media (OM) in the population of Greenland viewed in a historical and modern clinical perspective. Chapter 1 outlines the addressed problems and aims while chapters 2 and 3 deal with historical studies and an evaluation of the present knowledge based on the literature. Physical anthropological studies, using skeletal samples of adult Eskimo crania from before and after the colonization of Greenland in 1721 and information about modern living Eskimos (Inuit), have shown that OM sequelae of the temporal bones were significantly less common in pre-colonization Eskimos and that the mean area size of the pneumatized cell system in the temporal bone was significantly larger in pre-colonization Eskimos. These findings indicated an increase in OM after the colonization most likely caused by the social, cultural, habitary, and dietary changes due to increased contact with the outside world. Historical reports after the colonization confirm a high prevalence of OM especially in children. Modern epidemiological studies from the 1960's to 1980's in the Arctic region of Alaska, Canada, and Greenland along with reports from visiting consultant otologists in Greenland almost uniformly mention prevalent OM problems in children as well as in adults. The aim was therefore to further describe the epidemiological pattern of the different OM disease entities (acute OM (AOM), chronic OM (COM), COM with suppuration (CSOM), secretory OM (SOM), and cholesteatoma) and investigate the potentially associated risk factors in especially Greenlandic children because these diseases are primarily established and problematical in childhood. Chapter 4 describes the definitions used in the thesis and chapter 5 describes the studies included. Section 5.1 describes a study of cholesteatoma in Greenlanders. The study revealed an almost similar incidence of hospital treated children with cholesteatoma (6.6 per 100,000) as seen in comparable studies from other parts of the world. Furthermore, childhood cholesteatomas were the most aggressive. The frequency of residuals or recurrences after otosurgical treatment was high with a trend for better results when using the extensive canal wall-down procedure. It could be concluded that these patients urgently need close follow-up for at least five years postoperatively, if not lifelong. Section 5.2 describes a hearing screening survey of 167 school children using school registration charts. A high prevalence of hearing loss (HL) was found. A total of 43% of the children had hearing thresholds exceeding 20 dB at one or more frequencies between 250-8000 Hz in one or both ears, and 19% had the same type of HL in the frequencies 500-2000 Hz. HL was significantly associated with episodes of OM. These findings were in accordance with reports from Alaska and Canada. It is therefore concluded that a hearing screening programme of school children is important and that OM seems to have an impact on hearing in school children in Greenland. In section 5.3 an epidemiological survey is described concerning the prevalence of the different OM disease entities. The survey was carried out in Nuuk and Sisimiut and involved 740 children aged 3, 4, 5, and 8 years. A total of 591 children participated and selection bias was not found when controlling for age, sex, and episodes of AOM. The survey revealed that 52% of children in Nuuk and 54% in Sisimiut had some kind of pathological affection of their middle ear. COM and CSOM were found in 9%, but more prevalent among children in Sisimiut (12%) than in Nuuk (7%). Middle ear effusion (MEE) diagnosed by tympanometry was found in 23% in Nuuk and 28% in Sisimiut while simple tubal dysfunction (STD) was found in 13% and 8%, respectively. MEE and STD were associated with young age. Sequelae of OM was apparent in 11% in both towns. When comparing the results with a 10-year-older, almost similar survey of 142 children, it was evident that the OM situation had not changed in the period between the studies. The survey underlines the need for increased focus on the different OM entities in Greenlandic children. Section 5.4 deals with microbiological aspects. The nasopharyngeal microflora and ear discharge microflora of potential pathogens were evaluated in 54 children with AOM and in 201 control children without AOM. Very high carriage rates expressed qualitatively and semiquantitatively of potentially pathogenio bacteria were found in the nasopharynx of children with AOM (98%) but also in that of the control children (91%) and even in children denoted as being very healthy (94%). However, the same bacterial species were cultured from the nasopharynx and ear discharge as in comparable studies world-wide. Only S. pneumoniae was carried significantly more often in the nasopharynx of AOM children compared with age matched control children. Chlamydiae, M. pneumoniae, adenovirus, respiratory syncytial virus, parainfluenza- type 1, 2, and 3 virus, and influenza- type A and B virus were not major pathogens. In contrast, entero- and rhinoviruses were detected significantly more frequent in nasopharyngeal specimens from AOM children (59%) compared with age matched controls (33%) and also in 29% of the examined ear discharge specimens. It is therefore concluded that the potentially pathogenic bacterial load is early and massive. This alone or in interplay with entero- and rhinovirus infection and occasionally with other viruses may play an important role in the high prevalence of OM among children in Greenland. Section 5.5 deals with an examination of potential risk factors for AOM, recurrent AOM (rAOM), and COM in the same 591 children as studied in section 5.3. Early age at first AOM episode was associated with rAOM episodes (> or = 5 episodes since birth). Thus, the relative risk of developing rAOM was eight times higher if the first episode of AOM occurred before 7 months of age than after 24 months of age. Furthermore, compared with studies elsewhere in the world, a high proportion (40%) of the children in this survey had their first AOM episode during their first year of life and 41% of these children developed rAOM. It was also found that children had an increased risk of AOM, rAOM, or COM when both parents were born in Greenland, when parents also have had OM, when living in very crowded households, and when having experienced a long period of exclusive breast feeding, or when recalling of breast feeding was not possible. Gender, type, and size of housing, insulation standard of housing, daycare, exposure to passive cigarette smoking, and dietary habits were not associated with AOM, rAOM, or COM in the surveyed children. It is concluded that early onset of AOM occurs frequently in Greenlandic children and that a high proportion of these children develop rAOM. The study confirms that AOM is a highly multifactorial disease determined by a number of genetic and environmental factors. Finally, section 5.6 is a hypothesis generating study attempting to explain the high prevalence of early episodes of AOM in community-based children in Nuuk. The hypothesis is based on a possible association between findings of mannose-binding lectin genotypes, early Epstein-Barr virus infections and episodes of AOM, rAOM, or nasopharyngeal colonization with potentially pathogenic bacteria. However, the study does not support any of this hypothesis. In chapter 6, future studies are suggested and chapter 7 presents concluding remarks. PMID- 11725623 TI - Study on the secondary structure of protein in amide I band from human colon cancer tissue by Fourier-transform infrared spectroscopy. AB - BACKGROUND: Fourier-transform infrared spectroscopy (FT-IR) is a powerful tool for study of the secondary conformation of intact protein in biological tissues and for diagnosis of disease states. The purpose of this study is to utilize FT IR as a diagnostic tool to estimate the secondary structure of amide I in normal and cancer tissue of human colon. METHODS: Pairs of tissue samples from colon cancer and histologically normal mucosa tissue 5-10 cm from the tumor were obtained from a patient who underwent partial colectomy. In the present study we investigated the changes in the secondary structure of amide I of normal and malignant human colon tissue using FT-IR microspectroscopy with attenuated total reflectance. Secondary-derivative analysis relative to the original and curve fitting analysis infrared spectra were also carried out. RESULTS: The conformational changes of the secondary structure of protein in normal and cancerous human colon tissue were (1) the composition of alpha-helixes increased in human cancerous colon tissue (15.30%) compared to normal colon tissue (11.72%); (2) beta-sheets which were 46.8% in healthy tissue decreased to 30.71% in cancer tissue; (3) beta-turns increased from 26.28% in normal tissue to 38.48% in cancer tissue; and (4) the composition of random coils was not significantly different between normal tissue (15.12%) and cancer tissue (15.51%). CONCLUSION: Our data demonstrate that protein in colon cancer displays abnormal FT-IR spectra. The conformational changes of the secondary structure of this protein in terms of alpha-helixes, beta-sheets and beta-turns were observed in normal and cancerous tissue from the same individual were compared. PMID- 11725624 TI - Estimating stature from knee height for adults in Taiwan. AB - BACKGROUND: Many of the indices utilized in the assessment of nutritional status are dependent upon stature. Knee height has been used to predict stature in western countries when standing height cannot be measured, however, an adequate stature-knee height equation has not been established for the population in Taiwan. METHODS: A total of 1179 healthy men (603) and women (576) were recruited from those who had undergone comprehensive check-ups at their own expense from March through August 1998 at Lin-Kou Chang-Gung Memorial Hospital, Taiwan. The subjects were freely ambulatory without kyphosis or scoliosis noted on their chest X-ray or KUB films. The automatic supersonic system was used to measure stature and body weight. A Ross caliper was used to measure recumbent left knee height. Multiple linear regression was used to obtain equations to estimate stature using knee height and age. RESULTS: Stature of adults highly correlated with their knee height (r = 0.73 with 95%CI between 0.69 and 0.77 for men, and r = 0.68 with 95%CI between 0.63 and 0.72 for women) but declined with age (men: r = -0.17 with 95%CI between -0.09 and -0.24; women: r = -0.22 with 95%CI between 0.14 and -0.29). Knee height changed very little with age in both genders. To predict stature, a linear regression model should include knee height and age as predictors to reach r2 as 0.61 and 0.58 for men and women, respectively. The stature-knee height equations were "stature = 85.10 + 1.73 x knee height -0.11 x age" for men, "stature = 91.45 + 1.53 x knee height -0.16 x age" for women. CONCLUSION: Our stature-knee height equations provided good predictions on stature of adult population in Taiwan, and was superior when compared with equations developed from other populations. PMID- 11725625 TI - Auditory brainstem evoked potentials in healthy full-term and pre-term infants. AB - BACKGROUND: Auditory brainstem evoked potentials (ABEP) are used to evaluate the integrity of auditory pathways. Since there are difficulties in the performance and still no reference ranges for pre-term infants in Taiwan, the purpose of this study was to evaluate the characteristics of ABEP in full-term and pre-term infants. METHODS: Forty full-term and 52 pre-term infants were enrolled between January and December 1997. ABEP study was performed at 3-7 days after birth, 3, 6 and 12 months of age in the full-term group while the pre-term infants were assessed at 34-36 weeks of gestational age, at term age, and at 3, 6, 12 months of corrected age. Acoustic stimulation of mixed frequency rarefaction clicks was presented to each ear through earphones. The clicks were presented at an intensity of 70-90 dB (hearing level) and a gradual decrement of intensity to 20 40 dB to determine the threshold. Electrical activity was picked up by surface electrodes. Latency of peaks I, III, V, inter-peak latency I-III, III-V, I-V and the threshold were assessed. RESULTS: In the full-term group, the peak and inter peak latencies decreased with age. In the pre-term group, the responses of 70 dB stimulus intensity were 53.8%, 58%, 82.8%, 88.9%, 100% in the sequence and the threshold of response decreased with age. CONCLUSION: There are maturational changes after birth in both full-term and pre-term infants. We conclude that ABEP study is a sensitive, non-invasive tool for the evaluation of auditory pathway in infants. PMID- 11725626 TI - Serum cholesterol levels in mood disorders associated with physical violence or suicide attempts in Taiwanese. AB - BACKGROUND: Many previous studies have discussed the relationships between serum cholesterol levels and depression, violence and suicide in Western countries. In this paper, the relationships between serum cholesterol levels and mood disorders, physical violence and suicide attempts in Taiwanese psychiatric inpatients were reported. METHODS: A review of medical charts over a 1-year period was carried out in a population of 213 Taiwanese psychiatric inpatients that included 61 patients with affective disorders. The collected data included age, body weight, height, and serum cholesterol levels. RESULTS: There were no significant differences in the serum cholesterol levels between patients with mania and major depression, between patients with and without physical violence, or between patients who had and had not made a suicide attempt using analysis of covariance after age or body mass index adjustment. CONCLUSION: Due to the limitations of case numbers and study methods, the results need to be further clarified with larger numbers of patients and in controlled studies. PMID- 11725627 TI - Body weight, serum uric acid and lipid profile one year after spinal cord injury. AB - BACKGROUND: Survey of the serum lipoprotein profile, uric acid levels and body weight in Chinese patients with spinal cord injury (SCI) one year after injury, and to report the data of patients with SCI at different levels. METHODS: Seventy seven Chinese patients with SCI categorized into cervical SCI, thoracic SCI, and lumbar SCI groups were enrolled in the study. The serum uric acid and lipoprotein profiles of these patients were evaluated one year after injury. All of the data in those different groups of SCI patients were compared with normal levels. Body weight before and after injury were also evaluated. RESULTS: Serum uric acid was within normal limits in all three SCI groups. A similar lower in serum high density lipoprotein cholesterol (HDL-C) level was observed in all three groups of SCI patients. Higher serum levels were observed in total cholesterol (TC) in the lumbar SCI group, triglyceride and very low density lipoprotein cholesterol in the thoracic and lumbar SCI group, and TC/HDL-C ratio in all three SCI groups. Decrease of body weight were noted in cervical and thoracic SCI patients. CONCLUSION: Further long-term follow up was necessary to determine the serum lipid and body weight change in patients with SCI at different levels. PMID- 11725628 TI - Minimal change nephrotic syndrome associated with malignant thymoma: case report and literature review. AB - We report on a patient with nephrotic syndrome with myasthenia gravis and malignant thymoma status post thymectomy and radiotherapy. This 44-year-old woman underwent a thymectomy and localized radiotherapy for invasive thymoma in 1991. She also took azathioprine and pyridostigmine regularly for the control of her symptoms of myasthenia gravis following thymectomy. Neither evidence of recurrence of myasthenia gravis, nor enlargement of residual thymoma was noted following treatment in 1991. Unfortunately, this patient developed a pronounced nephrotic syndrome in 1999, a renal biopsy revealing a minimal-change glomerulonephritis as being present. The patient entered remission subsequent to steroid and cyclosporin therapy, whereas many previously-reported cases of a similar nature either died or remained with a persistent proteinuria. Herein, we review the literature pertaining to examples of nephrotic syndrome for those patients diagnosed with malignant thymoma, and discuss the possible mechanisms for this association, and emphasize the need for the aggressive treatment of the condition, recommending the early use of steroids and cyclosporins. PMID- 11725629 TI - Solid variant of papillary renal cell carcinoma. AB - Solid variant of papillary renal cell carcinoma is a newly recognized renal epithelial neoplasm. We encounter a case of a 36-year-old male patient with a solitary right renal tumor. The tumor is composed of solid sheets of small uniform cell with intervening micronodules, resembling the glomeruloid bodies in a metanephric adenoma. Abortive papillae rather than typical papillary formations are found in the micronodules. Furthermore, the cells in these micronodules demonstrate more cellular atypia with abundant eosinophilic cytoplasm comparing to the intervening cells. Both types of neoplastic cells are immunoreactive for epithelial membrane antigen, cytokeratins 7 and 17, but not vimentin and 34 beta E12. The features of the tumor are consistent with solid variant of papillary renal cell carcinoma. Solid variant of papillary renal cell carcinoma is similar in certain morphological and genetic features to metanephric adenoma, suggesting that they might be related. PMID- 11725630 TI - Risperidone augmentation of specific serotonin reuptake inhibitors in the treatment of refractory obsessive-compulsive disorder: report of two cases. AB - The efficacy of specific serotonin reuptake inhibitors (SSRI) in the treatment of obsessive-compulsive disorder (OCD) has been established, but more than 40% of patients continue to have a poor response to SSRI. Low-dose antipsychotic augmentation of SSRI has contributed to the amelioration of symptoms in the treatment of refractory obsessive-compulsive disorder patients. Due to the risk of side effects from traditional antipsychotics, atypical novel antipsychotic augmentation of SSRI may be a good choice in the treatment of refractory obsessive-compulsive disorder. Herein we report our experience with two OCD patients, one with a poor response to fluoxetine 80 mg per day for 3 months, and the other with poor response to fluoxetine 60 mg per day for 3 months. Both OCD patients also proved to have a poor response to paroxetine 60 mg per day for 6 weeks. Neither patient had received any psychotherapy or behavioral therapy. Both patients showed significant improvement in their obsessive-compulsive symptoms as measured by the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS), after risperidone 1 mg per day was added to their on-going use of paroxetine 60 mg per day. Within 4 weeks of adding risperidone, the two patients' Y-BOCS scores had decreased by 57% and 53%. This result suggests that risperidone augmentation of SSRI may be a good choice and an effective treatment strategy for refractory OCD. PMID- 11725631 TI - Epithelial growth within the lamellar interface after laser in situ keratomileusis. AB - We report on a case of 36-year-old man with excessive myopia and amblyopia who underwent laser in situ keratomileusis (LASIK). The surgery was performed smoothly, and visual acuity improved greatly. The patient was lost to follow-up in our clinic until 2 months later when he returned because of blurred vision. Epithelial growth within the lamellar interface was noted which disclosed white, plaquelike, confluent opacities within the lamellar interface on the visual axis. Topography showed an irregular astigmatism overlying the entrance to the pupil. The flap was lifted at the flap edge, but it was difficult to lift it around the area of epithelium growth within the lamellar interface under an operating microscope. The epithelial tightly adhered to the lamellar interface, so surgery was aborted because of the possibility of flap perforation. Prednisolone acetate 1% was used for one month, but the epithelial growth within the lamellar interface persisted. LASIK has the impressive potential to correct refractive errors. Epithelial growth within the interface after LASIK, although usually a benign complication, can lead to a poor outcome and even permanent vision loss. PMID- 11725632 TI - Zona-free oocyte fertilized with intracytoplasmic sperm injection and underwent further division: case report and literature review. AB - The zona pellucida (ZP) plays a protective role during fertilization and early embryonic development. It is related to sperm binding, the acrosome reaction, prevention of polyspermic fertilization, and holding blastomeres together before the morular stage. Zona-free oocytes are accidentally encountered. If these oocytes are healthy, they can be fertilized normally by intracytoplasmic sperm injection (ICSI). We reported on a couple with male infertility undergoing oocyte retrieval after ovarian hyperstimulation. Before the ICSI procedure, cumulus cells surrounding the oocytes were removed, which resulted in one oocyte escaping from its ZP. The zona-free oocyte was fertilized normally with ICSI and developed to the 8-cell stage. We observed that the zona-free zygote had the ability to further divide, despite its loose contact. The zona-free embryo was transferred with other zona-intact embryos, but the implantation failed. We conclude that zona-free oocytes can be rescued, fertilized with ICSI, and cultured for further transfer or cryopreservation. PMID- 11725633 TI - [Development of donor cells in chicken-duck chimeric embryos]. AB - Chicken W chromosome-specific DNA fragments were obtained after transformation, extraction, purification and digestion with EcoR I of pUGD1201. The W chromosome specific DNA probe was a 1.2 kb insert, which was labelled by the random technique using a Digoxigenin Labeling Kit. Chicken PGCs from a line of White Leghorns were microinjected into Maya duck blastoderms at stage X. The embryos at 6,8,9,12,16,18 days of incubation were used to study the development of chicken PGCs in the chimeric embryos. Used chicken W chromosome-specific DNA probes by in situ hybridization, development of donor cells in chicken-duck chimeric embryos were studied. In 54 chimeric embryos, there were different degrees of chimerism in the organs. The chimeric rate of the livers was the highest, which of the gonads was the lowest. It is demonstrated that somatic and germline chimeras can be yielded successfully by transferring stage X chicken blastodermal cells. PMID- 11725634 TI - [Genetic analysis of gelatinization temperature in rice via microsatellite(SSR) markers]. AB - Rice gelatinization temperature is an important character contributing to cooking quality. Here the inheritance of gelatinization temperature (GT), which was represented by alkali spreading value (ASV), was reported. Two parents, Balilla (japonica variety) and Nantehao (indica variety), which were significantly different on GT-ASV, were selected to construct a backcross population Balilla/Nantehao//Balilla containing 142 individuals. And ASV was investigated in the population, a continuous distribution with two obvious peaks was observed. It indicated that GT-ASV was controlled by one major gene, also modified by some minor genes. In order to map the major and minor genes and estimate the effects of genes. A total of 119 SSR markers were employed to construct a linkage map; further a genome-wide detection was carried out by interval mapping method. The results showed that 6 QTLs were detected, of which, qASV6-1 located on chromosome 6 was a major gene with 87.6% variance explained, and alleles from parent Nantehao could decrease the value. It shoud be the same locus as the alkali degeneration gene (alk). The other QTLs (qASV2, qASV3, qASV6-2, qASV9, and qASV11) all belong to minor genes, which were located on chromosome 2,3,6,9 and 11, respectively. In two parents, they carried the positive and negative alleles simultaneously. These results will be helpful for rice quality breeding and improvement. PMID- 11725635 TI - [Studies of the integration and expression of exogenes in transgenic rice obtained via particle bombardment transformation]. AB - Various integration events from simple to complex have been found in transgenic elite rice varieties, which were co-transformed with bar and cecropin B gene by particle bombardment after system studies of the structure and expression of the transgene. It was displayed that there were one to seven insertion loci and one to ten copies of transgene in different transformants. No absolute correlation was found between the numbers of transgene copies and the gene expression. Different level expression of transgenes with similar insertion mode was also discovered. The expression cassette integrity and transcription frequency of selectable bar gene were more than non-selectable cecropin B gene, whereas gene silence in plants integrated with intact bar gene was found and higher expression level of cecropin B gene with 0.6 kb fragments degraded in terminator was deduced. PMID- 11725636 TI - [Analysis of heading time genotype for a rice male sterile line Zhenshan 97A]. AB - Zhenshan 97A, a rice male sterile, were applied widely since the release of hybrid rice in 1973 in China, but the genotype of heading time in this sterile line was still unknown. This definitely limited the further use of this sterile line in breeding practice and re-production of hybrid seeds. To solve this problem, we analyzed the segregation pattern of phenotype of heading time in progenies from crosses between Zhenshan 97A and four tester cultivars, Akihikari (e1e1e2e2e3e3 Se-1eSe-1e), Koshihikari (E1E1E2E2e3e3Se-1eSe-1e), Nipponbare (E1E1e2e2e3e3Se-1Se-1) and Hinohikari (E1E1E2E2e3e3Se-1Se-1), whose genotypes of heading time were already known. The results showed that the genotype of heading time in Zhenshan 97A was e1e1e2e2E3E3Se-1Se-1 and it also carried a recessive inhibitor i-Se-1 for phtoperiod-sensitivity. Meanwhile, a major photoperiod sensitive dominant genes Se-1 and other modified photoperiod-sensitive genes: i Se-1, E3, Hd3(En-Se-1), Hd5 and Hd6, were identified in Zhenshan 97A by crossing with QTL nearly isogenic lines: NIL (Hd1), NIL (Hd2), NIL(Hd3), NIL(Hd5) and NIL(Hd6). PMID- 11725637 TI - [Molecular markers linked to dwarf gene Rht3 in wheat]. AB - The dwarf gene Rht3 derived from Tom Thumb variety, a Tibetan wheat of China, is a dominant gene with the insensitivity to gibberellic acid. Rht3 shows to depress alpha-amylase activity and hence enchance the resistance to preharvest sprouting mainly through its effect on the expression of alpha-Amy1. Near isogenic lines with Rht3 and their segregating population were analyzed by PCR and RFLP techonology. In RAPD analysis, out of 310 random primers (10 bp) screened, only three primers of UBC389, OPV-06 and S1060 revealed polymorphisms in NIL from 310 random primers. Fragments S1060(1900) and S1060(2000) amplified by primer S1060 were shown to be linked to Rht3 with a genetic distance 7.1 cM and 9.2 cM. Fifty three probes specific for short arms of homoeologous group 4 were screened in RFLP analysis. Xpsr584, XksuF8 and Xcdo38 showed polymorphisms between the NIL. The linkage analysis indicateded that Xpsr584 was linked to Rht3 with a genetic distance 8.0 cM. PMID- 11725638 TI - [Molecular tagging of a major powdery mildew resistance gene MlG in wheat derived from wild Emmer by using microsatellite marker]. AB - Powdery mildew caused by Erysiphe graminis f. sp. tritici is one of the most important wheat diseases in many regions of the world. Breeding for resistant cultivars has been proved to be an effective and environmentally safe method to control diseases in wheat production. It is necessary to search for more resistance genes for the diversification of resistance genes in wheat breeding. An Isreali wild emmer wheat (Triticum dicoccoides) accession "G-305-M" was found resistant to the prevailing E. graminis f. sp. tritici isolate Race No. 15 in Beijing region. The powdery mildew resistance has been transferred from G-305-M into common wheat by crossing and backcrossing (G-305-M/781//Jing 411* 3). Genetic analysis showed that the resistance was controlled by a single dominant gene at the seedling stage. A segregating BC2F3 family of the cross "G-305 M/781//Jing 411* 3" with 167 plants was chosen for SSR analysis. Totally 96 wheat microsatellite primer pairs were screened, only one primer pair WMS570 could generate polymorphic DNA fragments between the resistant and susceptible plants. After evaluating this polymorphic marker in the segregating population, the microsatellite locus Xgwm570 mapped on chromosome 6AL was found to be linked to the resistance gene, with the estimated genetic distance of 14.9 +/- 3.0 cM. Based on the origin and chromosomal location of the gene, it is suggested that the resistance gene derived from G-305-M should be a novel Pm gene and is temporarily designated MlG. PMID- 11725639 TI - [Genetic diversity detected by DNA markers and phenotypes in Upland cotton]. AB - Genetic diversity of Upland cotton (Gossypium hirsutum L.) cultivars was evaluated by SSR, ISSR, and RAPD. The genomic DNAs of 36 domestic and exotic cultivars were amplified with 81 SSR primer pairs; 7 ISSR primers and 53 RAPD primers, which yielded 282 polymorphic bands. The range of pair similarity coefficient (Nei, M. And W. Li 1979) between cultivars is from 0.5745 to 0.9291. The result indicated that although the present cultivars have genetic variation to a certain extent, the genetic basis of them is relatively narrow, and it is essential to find some new germplasm for the development of new cultivars. As expected, the classification of the cotton cultivars using the Unweighted Pair group Method with Arithmetic Average based on the pair similarity coefficients is mostly consistent with known pedigree information and breeding experience. The 36 cotton cultivars have been mainly classed into exotic cultivar group, Chinese Xinjiang ecological region cultivar group, early-mature domestic cutivar group and mediate-mature domestic cultivar group. Carmen and Colorado cultivars from Spain have the lowest similarity with other cultivars, while TM-1 (genetic standard line), MD5lne and PD9364 cultivars from American have higher similarity with domestic cultivars. The reason may be that the parents of the domestic cultivars were mainly introduced from America. However, clusters of domestic cultivars are not all in accordance with their ecological regions, which may result from the pedigree complexity of the cultivars. Meanwhile the genetic diversity of 36 cultivars was investigated at phenotypic level with agronomic and fiber quality traits in the field experiments in 1999 and 2000, and was measured by Euclidean distance on the genotypes. Although grouping result of cluster analysis based on the Euclidean distance is similar than that of molecular data with a significant negative correlation of -0.3350 between the pair similarity coefficients and the Euclidean distance, the subgroup classification of the two methods gives different result. The molecular markers can be effectively applied in classifying cultivars within the group. The study provides useful information for the selection of parents in the development of cotton cultivars and hybrids as well. PMID- 11725640 TI - [Construction and analysis of a genetic linkage map of soybean]. AB - A high-density genetic map was constructed with a RILs population (NJRIKY) from a cross of two cultivated soybean Kefeng 1 and Nannong 1138-2, using RFLP, SSR, RAPD and AFLP markers. The map consists of 24 linkage groups, which include 792 genetic markers, and covers 2,320.7 cM with an average genetic distance of 2.9 cM. The higher polymorphsims between two parents were identified with SSR markers. Due to its simplicity and stability, they could be used as anchored marker to integrate different maps constructed with different genetic markers. AFLP marker system is efficient for map construction while it easily form clusters to cause big gaps in the map. A total of 21.7% of markers was found with distorted segregation in the RILs population. The high-density molecular linkage map is fundamental for gene localization, comparative genomics, and QTL mapping of important agronomic traits. PMID- 11725641 TI - [Two complementary dominant genes controlling apomixis in genus Citrus and poncirus]. AB - Apomixis is very common in Citrus genus and its related genus. Two monoembryonic tangerine varieties (C. reticulata Blanco) Clementine and Wilking were used as seed parents to cross with four polyembryonic orange varieties [C. sinensis (L.) Osbeck]. Both sexual and apomictic progenies were found in each F1 population with different segregation ratios. In a total of 74 Wilking progenies, 23 were monoembryonic and 51 polyembryonic and the ratio of sexual and apomictic progenies was about 1:2 tested either in each individual cross or in the whole Wilking cross combionations. In Clementine progenies, 84 were monoembryonic and 71 polyembryonic and the ratio was about 1:1 tested either in each individual cross or in the whole Clementine cross combionations. According to the results together with other data published, a possible apomixis controlling mechanism was proposed, which involves two complementary dominant genes named as A1 and A2 that control apomixis in genus Citrus and Poncirus. Trees of genotype A1-A2-, except for homozygous of dominant gene A1 (which is lethal), can produce apomixis seeds. And those of other genotypes will produce sexual seeds. The segregation and recombination of these two genes accorded with Mendel's genetic laws. The proposed mechanism could explain genotypes controlling polyembryony-monoembryony existing both in nature species and artificial hybridization progenies as well as most of the known hybridization results. PMID- 11725642 TI - [Screening, cloning and sequence analysis of the gene for pyruvate kinase from Achlya bisexualis cDNA library]. AB - The gene for pyruvate kinase, which is involved in glycolysis and catalyze the biochemical reaction from phosphoenolpyruvate to pyruvate, was screened, cloned and sequenced from non-photosynthetic Oomycetes, Achlya bisexualis cDNA library. The size of the gene is about 1.6 kb. The amino acid sequence of the gene was deduced using the GCG program. On basis of analysis by the on-line PSORT II program the deduced amino acid sequence showed that the pyruvate kinase expressed by the gene was located in the cytosol. This result indicated that it is possible that the process of glycolysis for the lower form of eukaryotic cell such as Achlya bisexuali may occur in cytosol, which is different from Liaud's report. PMID- 11725643 TI - [Bioinformatics analysis of autophagy 5 gene structure]. AB - In order to study the relationship between autophagy and apoptosis, APG5 gene structure was revealed by bioinformatics analysis and meantime a new isoform resulted from alternative splicing of the hAPG5 gene was confirmed, which was hereby designated as human autophagy 5 beta (hAPG5 beta; LOCUS AF293841, GenBank). We cloned and sequenced the cDNAs from fetal brain and B cell cDNA libraries using the known hAPG5 cDNA open reading frame sequences as primers. The cDNA obtained from the human fetal brain cDNA library was identical to the known hAPG5 cDNA. However, the cDNA from adult brain cDNA library was 129 bp shorter in length, lacking the sequence corresponding to those from positions 434 to 563 of the hAPG5 cDNA. Through search in public database and sequence comparisons and assembly 4 related sequences, APG5 genomic sequence was obtained. We found that the hAPG5 gene had 8 exons, and those sequences missing in hAPG5 beta cDNA exactly corresponded to exon 3. By bioinformatics software, the structure of introns, exons, splicing sites, promoter and polyA were demonstrated. Moreover, we were able to express both hAGP5 and 5 beta cDNA clones in human hepatocytes and HeLa cells using pEGFP-C1 vector. In conclusion, our data indicate that a systematic bioinformatics method of finding protein diversity from alternative splicing is a good approach in post-genome biology. PMID- 11725644 TI - [Genetic polymorphisms of 9 STR loci in Achang ethnic group in Yunnan Province]. AB - Blood samples were collected from the unreleatled individuals in the Achang ethnic group in Yunnan Province. Genetic distribution for nine STR loci and amelogenin locus were determined in Achang ethnic group based on GeneScan. 96 samples were denatured of gel electrophoresis. The databanks in Achang ethnic group were generated by using GeneScan, genotyper, and genetic distribution analysis. 69 alleses and 166 genotypers were observed, with the corresponding frequency being 0.0050-0.6100 and 0.0100-0.3900. The average H is 0.7381. The comulated DP is 0.9999999. The comulated EPP is 0.9999989. The allele distribution of the loci was in good agreement with the Handy-Weibeng equilibrium. It is useful to establishing DNA databanks for studying gene natural resources, very valuable in the study of forensic science, anthropology and ethnic. PMID- 11725645 TI - [Linkage analysis and mutation detection of GRIA3 in Smith--Fineman--Myers syndrome]. AB - To determine the role of GRIA3 in the etiology of Smith--Fineman--Myers syndrome (SFMS), polymorphic short tandem repeats within GRIA3 gene were genotyped by PCR and denaturing polyacrylamide gel electrophoresis to test linkage between GRIA3 and the gene responsible for SFMS. The open reading frame of GRIA3 was detected for mutation by PCR amplification and direct sequencing in affected and normal males from SFMS family. One of the two short tandem repeats was informative in SFMS family. Tight linkage between SFMS locus and GRIA3 gene was established by STR3 within GRIA3 gene. No disease--causing mutation was found within the open reading frame of GRIA3 gene. The disease in SFMS family from Shandong (China) is not caused by the mutation within open reading frame of GRIA3 gene. PMID- 11725646 TI - [The study of P68 RNA helicase on cell transformation]. AB - The P68 nuclear protein is an ATP-dependent RNA helicase and RNA-dependent ATPase, and has been proved to have functions in the regulation of cell growth and division. Different phenotypes of P68 between tumouric cell lines (HeLa, TC3H10) and non-tumouric cell lines (NIH3T3 and NC3H10) were observed, and we reasoned that overexpression of P68 might profoundly affect cellular growth properties. We report here that overexpression human P68 by transfection of recombinant P68 expression vector in NIH3T3 and NC3H10 fibroblasts causes their tumorigenic transformation as determined by four criteria: (1) the morphology of transfectants from long fibric to spindle shaped; (2) formation of transformed foci on the monolayer of cells; (3) the transfectants acquired the ability to grow on soft agar forming cell microsphere cube; (4) tumor formation in nude mice after injection of transfected cells for 3-4 weeks. PMID- 11725647 TI - [Coding region cDNA sequence cloning of rat neuroglobin gene, its polymorphism feature and tissue expression profile analysis]. AB - The coding region cDNA sequence of rat neuroglobin (NGB) was obtained by RT-PCR technique using a degeneracy PCR primer pair based on previously reported cDNA sequence of human and mouse NGB gene. Result demonstrated that the coding region cDNA sequence of rat NGB gene is 456 bp in length, which could encode a protein of 151 amino acids. The rat NGB gene is highly homology with mouse (96%) and human (88%) NGB gene. However, several polymorphism sites were also detected in the rat NGB coding region: 113 t/c [L38P], 133 a/g [N45D], 388 a/g[R130G], 417 t/c. The cDNA sequence of rat NGB gene has been registered in GenBank under the accession number AF333245. Moreover, highly expression level of rat NGB in brain, liver, kidney, heart and skeletal muscle was detected by using multiple tissue RT PCR technique, indicating the functional importance of this novel gene. PMID- 11725648 TI - E-therapy: practical, ethical, and legal issues. AB - E-therapy is a term that has been coined to describe the process of interacting with a therapist online in ongoing conversations over time when the client and counselor are in separate or remote locations and utilize electronic means to communicate with each other. It is a relatively new modality of assisting individuals resolve life and relationship issues. E-therapy utilizes the power and convenience of the internet to allow simultaneous (synchronous) and time delayed (asynchronous) communication between an individual and a professional. For the purposes of this paper, e-therapy is defined as a licensed mental health care professional providing mental health services via e-mail, video conferencing, virtual reality technology, chat technology, or any combination of these. It does not include self-help methods such as public bulletin boards or private listservs. E-therapy is not psychotherapy or psychological counseling per se since it does to presume to diagnose or treat mental or medical disorders. However, e-therapy is flexible enough to also address many difficulties which clients present to the online therapist. As in other types of therapy, such as bibliotherapy, occupational therapy, and rehabilitation therapy), e-therapy does assist a person in addressing specific concerns with specific skills. This article examines the following issues of e-therapy. First, the types of e-therapy and related services are described to provide a background for the article. Second, the ethical codes which have been adopted by three major professional organizations (American Counseling Association, National Board for Certified Counselors, and the International Society for Mental Health Online) pertaining to e-therapy are summarized for professional and consumer use. Finally, the practical, ethical, and legal issues of e-therapy services are discussed fully. PMID- 11725649 TI - Virtual reality monitoring: phenomenal characteristics of real, virtual, and false memories. AB - This study explored virtual-reality (VR) monitoring, the decision process by which people discriminate memories of real and virtual events. In a study phase, subjects touched 10 real objects with their real finger and touched 10 virtual objects (visual only, no tactile feedback) with their cyberfinger in VR. One week later they took a real versus virtual versus new source identification test. After the source identification test, subjects rated phenomenal qualities associated with each memory, using a Virtual-Real Memory Characteristics Questionnaire (VRMCQ). For old items, results from the VRMCQ are consistent with the idea that VR monitoring draws on differences in qualitative characteristics of memories for perceived and virtual events/objects (consistent with Johnson and Raye). However, subjects also reported similar qualities associated with their false memories for new items, suggesting that they sometimes infer/reconstruct the qualities a memory should have, based on their decision of its source of origin. Furthermore, VR monitoring might prove useful as a sort of Turing test of how convincing the virtual world is, and the VRMCQ can identify which qualities of the virtual experience (e.g., color) require improvement. Examples of applications are discussed. PMID- 11725650 TI - Internet heavy use and addiction among Taiwanese college students: an online interview study. AB - This paper presents empirical qualitative results of Internet heavy-use and addiction among some college students in Taiwan. It offers in-depth, online interviews of student-subjects in order to facilitate an interdisciplinary understanding of Internet heavy use, addiction and its potential impacts. A total of 83 subjects were interviewed, both as individuals and in chat-room groups. The analysis of qualitative data presented in six major themes: (1) Internet use and reasons; (2) Internet features; (3) the Internet as replacement for other media; (4) impact of Internet overuse; (5) controlling Internet use; and (6) coping with Internet withdrawal. Discussions, explanations, along with examples and quotes from subjects, are provided in each section. Implications for student affairs administrators and further research directions are also addressed. PMID- 11725651 TI - Knowing me, knowing you: reciprocal self-disclosure in Internet-based surveys. AB - Candid self-disclosure is desirable for many behavioral science studies. Although there is ample evidence that self-disclosure is increased when people communicate or participate in research over the Internet, few studies have looked at ways of increasing this effect. In the present pilot study, participants were randomly allocated to either a condition in which they received self-disclosing information about the experimenter (and then moved on to the study) or were directed straight to the study. Participants completed six open response questions on the Internet. Participants who received the experimenter disclosure divulged a significantly higher quantity of information about themselves, but their answers were not scored as significantly more revealing or intimate than those participants who did not receive the experimenter disclosure. Implications for conducting research over the Internet are discussed. PMID- 11725652 TI - Internet relationships: people who meet people. AB - Current research suggests that the Internet has become a popular medium for forming interpersonal relationships. Not only are many people developing cyberfriendships, but some move beyond virtual communication and interact more directly through telephone contact and face-to-face encounters. This pilot study examined the personality characteristics of 30 participants (17 men and 13 women) who were regular Internet users and who had used the Internet to meet people mainly through chat rooms. The findings of this study suggest that people who use the Internet meet others tend to be truthful in general in their interactions although both men and women often did not reveal their true names. Eighty percent of the subjects formed casual or friendly relationships, whereas 6% formed intimate or romantic relationships. Approximately one-third of the subjects made some form of offline contact, with 40% talking on the telephone and 33.3% meeting face-to-face. Furthermore, examination of personality styles indicated that cyberfriends tend to be socially skilled, have strong verbal skills, and demonstrate empathy for others. Most subjects were careful about protecting their anonymity, and none of the subjects who met face-to-face did so without first talking on the phone. This suggests that individuals who make friends via the Internet tend to take appropriate precautions and find this medium an effective and safe way to interact with others and to expand one's social system. PMID- 11725653 TI - Child seduction and self-representation on the Internet. AB - This paper presents a case study of a man charged with the offense of downloading child pornography from the Internet. He had used the Internet to traffic child pornography, and, in addition, to locate children to abuse, to engage in inappropriate sexual communication with children, and to communicate with other pedophiles. Such offenses were facilitated by self-representing in Chat rooms as both a child and an adult. The case study illustrates how such offenders move through a repertoire of offending behavior and discusses the role that the Internet can play in supporting inappropriate and disinhibited sexual behavior that victimizes children through the trading of child pornography and possible child seduction. The Internet is seen to play a unique role in allowing individuals to self-represent aspects that might otherwise remain hidden or dormant. PMID- 11725654 TI - Women, guilt, and home computers. AB - This paper will discuss, with reference to a field study examining attitudinal, emotional, and physical barriers, some of the ways in which some women are currently restricted in accessing networked computers in their own households. The study was prompted by a concern that current discussions focussing on the potential of computer-supported distance learning do not acknowledge differences in women's experience of family and household which may impinge on access. It will be suggested here that the domestic arrangements of space and time with home computers are crucial factors in supporting women's access to learning and personal development via the Internet. PMID- 11725655 TI - Relation of statement length and type and type of chat room to reciprocal communication on the Internet. AB - This study examined the relation of statement type (neutral, questioning, provocative, or bizarre) and length of statement (short vs. long) to reciprocal conversation in Internet chat rooms. The results indicated that provocative statements had a significant impact on reciprocal communication in two out of three different types of Internet chat rooms. These findings support the hypothesis that the amount of reciprocal communication in a chat room is directly related to the type of statements made by the participants. The results also suggest that the type of statement that elicits the greatest reciprocal communication may vary with the type of chat room. PMID- 11725656 TI - Age/sex/location: uncovering the social cues in the development of online relationships. AB - Past research on online relationships has predominantly been concerned with how the quality of online relationships compares with offline relationships. This research has been more concerned with the medium itself than with the meanings that users construct around their interpersonal interactions within this medium. The current paper seeks to redress this imbalance by exploring the ways that available social cues are used to shape the meanings of online relationships. Sixty Internet users, ranging in age from 19-51 years, were interviewed about their online relationships. It was found that ideals that are important in traditional relationships, such as trust, honesty, and commitment are just as important in online relationships; however, the cues that signify these ideals vary. PMID- 11725657 TI - Are women occupying positions of power online? Demographics of chat room operators. AB - Internet statistics indicate a reduction in the gender discrepancy online. Yet, what is the situation within specific online communities like Internet Relay Chat (IRC)? Likewise, what is the gender status of those occupying positions of power online? An exploratory study of chat room operators (those who govern chat rooms) was conducted to investigate gender differences in operator's demographic characteristics and IRC experience. Whether those less satisfied with their real life occupation were attracted to chat room operator positions was also investigated. A survey of 423 chat room operators was administered, comprising 25% women. Real-life occupations of chat room operators covered a broad spectrum, from professional and managerial to service, sales, and production workers, as well as those not employed. The most common occupational category cited was student, with very similar proportions of men and women occupying high-status positions. Of the occupations listed, 23% fell within the IT industry, with significantly more male than female operators working in this area. Majorities of both genders were satisfied with their real-life occupation. There was no relationship between job satisfaction and IRC experience or time spent as chat room operator. There were no gender differences for IRC experience. Majorities of both genders had been using IRC for 1 to 3 years or more, used IRC daily, and spent most or all of their time on IRC as operators. Ages ranged from 11 to 66 years, with the mean age 25 years. Women were significantly older than men. A significant proportion of men and women were from North America. PMID- 11725658 TI - Does a quality product mean a quality restoration? PMID- 11725659 TI - The microtensile bond test: a review. AB - PURPOSE: The purpose of this review is to describe all of the various modifications of the microtensile bond test in one paper, so that investigators can select the modification that best suits their needs. METHODS: The essence of the microtensile test is the division of resin-bonded teeth into slabs between 0.5 and 1.0 mm thick that are then trimmed in such a manner that tensile force will be concentrated on the bonded interface during testing. Among the many advantages of the technique are that each tooth produces multiple specimens. Further, there is no need for a matrix to limit the bonded surface area, since the area is determined by the dimensions of the trimmed specimens. RESULTS: The various modifications of the microtensile test have been used to measure differences in regional bond strength across occlusal dentin, down the external surface of teeth from crown through roots, down the internal surface of root canals from cervical to apical thirds, as well as to compare normal vs caries affected occlusal dentin and normal vs sclerotic cervical dentin. The technique is ideal for evaluating the long-term durability of resin-hard-tissue bonds. CONCLUSION: The microtensile test methods offer versatility that cannot be achieved by conventional methods. It is more labor-intensive than conventional testing, but holds great potential for providing insight into the strength of adhesion of restorative materials to clinically relevant sites and substrates. PMID- 11725660 TI - Practitioner variability effects on dentin bonding with an acetone-based one bottle adhesive. AB - PURPOSE: The objective of this study was to investigate the effect of operator variability on the marginal performance of resin composite restorations bonded with an acetone-base one-bottle adhesive in standardized dentin preparations. MATERIALS AND METHODS: Five general practitioners were recruited for this trial. In the first group, each dentist received six extracted human teeth with dentin preparations (O 3.5 mm, 1.5 mm deep), cut in flatground proximal dentin, and original packages of Gluma One Bond adhesive and Charisma resin composite (Heraeus-Kulzer, Werheim, Germany) including instructions for use. For the second group, the operators were orally instructed, emphasizing the importance of the moist technique, before they received another six teeth for restoration with the same materials. Finally, a third group of six preparations was restored in order to evaluate a possible training effect. The restored teeth were stored in a hygrophor prior to removal of excess material and microscopic determination of the maximum marginal gap width (MGW). Statistical analyses were performed with the Kruskal-Wallis and Wilcoxon tests at p < 0.05. RESULTS: Apart from two samples, preparation margins in the first test group showed gaps between 1 and 12 microns. In the second and third groups, 15 and 16 restorations were gap free, respectively. Significant interoperator variation was found in the first group only. Marginal performance of group 1 restorations was significantly inferior to groups 2 and 3, which were not different (p = 0.79). CONCLUSION: Application of the one-bottle adhesive is technique-sensitive and requires meticulous attention to the instructions. PMID- 11725661 TI - Bonding procedures for intraoral repair of exposed metal with resin composite. AB - PURPOSE: To determine if tin plating can be recommended for intraoral repair of ceramic veneered cast restorations where metal of unknown composition is exposed by loss of ceramic. This study investigated the effectiveness of surface treatments incorporating tin plating and unfilled resin to enhance the tensile bond strength of a resin composite restorative to three different metal ceramic casting alloys. MATERIALS AND METHODS: Gold-platinum, palladium-tin and nickel chromium alloys were used to fabricate 120 rods of each alloy, 4 x 15 mm. The end of each rod was ground perpendicular on 600-grit SiC paper and grit blasted with 50-micron alumina. Rods from each alloy were divided into four groups of 30 to receive one of the following treatments before bonding pairs end to end with a visible light-polymerized resin composite (Herculite XRV): 1) direct bonding with the resin composite; 2) tin plating (Micro-Tin) and bonding; 3) application of unfilled resin (Chameleon) and bonding; 4) tin plated, application of unfilled resin and bonding. The bonded samples were stored in distilled water, incubated at 37 degrees C for 24 hours and tested for tensile bond strength at a crosshead speed of 1 mm/min in a Lloyd 1000R machine. The mode of failure was examined using a stereo zoom microscope. RESULTS: A statistically significant increase in tensile bond strength was demonstrated between the control (group 1) and both the gold and palladium alloy treated with tin plating and unfilled resin (group 4). Tin plating, or tin plating with the application of unfilled resin had no statistically significant effect on the tensile bond strength of resin composite to the nickel-chromium alloy. CONCLUSION: The results demonstrate that tin plating, in conjunction with the application of a low-viscosity unfilled resin, optimizes the tensile bond of a resin composite to the three alloys used in the study. This procedure can be recommended for intraoral repair of exposed metal when the type of alloy belongs to one of the investigated alloy groups. PMID- 11725662 TI - A two-year randomized, controlled clinical evaluation of bonded amalgam restorations. AB - PURPOSE: The clinical performance of adhesively bonded dental amalgam restorations was compared with that of traditionally placed non-bonded control amalgams in a randomized prospective study. MATERIALS AND METHODS: One-hundred thirteen Class II Dispersalloy amalgams were placed in permanent molars and premolars of 31 (21 males, 10 females) human subjects with their informed consent. By random assignment, 60 amalgam restorations were adhesively bonded using ED Primer and Panavia 21 TC (both Kuraray Co, Japan), and 53 traditional non-bonded restorations were placed. Most preparations involved replacement of defective amalgam restorations. Clinical recalls were conducted by experienced evaluators using modified USPHS criteria for occlusal and proximal anatomic form, occlusal and proximal marginal adaptation, and occlusal and proximal surface roughness. RESULTS: For the categories of anatomic form, marginal adaptation, surface quality, and temperature sensitivity, there were no significant differences between bonded and non-bonded amalgam restorations (chi-square analysis, p > 0.05). Three non-bonded restorations were lost at 4, 7, and 24 months from preparations with no deliberate retention. CONCLUSION: After two years' clinical service, there were no failures among the amalgam restorations adhesively bonded using ED Primer and Panavia 21 TC, but three non-bonded restorations failed due to lack of retention. For traditional preparations, adhesively bonded amalgams of the type investigated perform as well as non-bonded amalgams over two years' clinical service. PMID- 11725663 TI - A three-year evaluation of restorations placed with a smear-layer-mediated dentin bonding agent in non-carious cervical lesions. AB - PURPOSE: To evaluate the handling characteristics, clinical behavior, and effectiveness of Coltene ART Bond/Brilliant restorations in mixed Class V preparations in maxillary premolar and canine teeth of adult patients over a three-year period. MATERIALS AND METHODS: Single-center, uncontrolled, clinical evaluation of 50 restorations in 26 patients extending over 36 months. RESULTS: At three years, 34 restorations, or 68% of those originally placed, in 20 patients were available for review. One of the 34 restorations had been lost 15 months after placement. In terms of marginal integrity and staining, anatomic form, surface texture and staining, and postoperative sensitivity, over 90% of the restorations reviewed at three years were rated alpha. Following difficulties in obtaining excellent color match in relation to 14 (28%) of the restorations at the time of placement, only 44% of the restorations were rated alpha at three years. CONCLUSION: It was concluded that with the possible exception of excellent color match, ART Bond/Brilliant restorations may be found to perform favorably in mixed Class V lesions in selected adult patients over a period of at least three years. The findings indicate that resin composite restorations placed with a smear-layer mediated dentin bonding agent may be found to have a favorable clinical outcome in non-carious cervical lesions. PMID- 11725664 TI - Intentional pulp capping: a clinical perspective of the adhesive experience. AB - A new standard of biocompatibility has evolved for dental tissues. This paper attempts to trace the extraordinary transformation of composite resin from dissident to healer. Intentional pulp capping is a logical progression of the adhesive era given the fact that current enamel-dentin bonding systems are biologically acceptable and potentially provide a gap-free therapeutic seal at the restorative interface. The probationary status of the adhesive pulp capping procedure is acknowledged; nevertheless, it is abundantly clear that there is a place for this treatment modality in clinical practice. PURPOSE: The objective of the accompanying pictorial essay is to give a clinical perspective of the author's experience with intentional pulp capping over the past seven years. The focus of this paper is the detailed examination of the clinical variables which influence the quality of the "seal" surrounding the exposed pulpal tissues. PMID- 11725665 TI - 3M meeting. Bandol, 20-21 September 1999. PMID- 11725666 TI - The price of progress. PMID- 11725667 TI - The pioneers of enamel and dentin bonding. PMID- 11725668 TI - Dentin bonding--questions for the new millennium. AB - PURPOSE: The objective of this review article is to summarize the most recent concepts in dentin bonding and, simultaneously, challenge some convictions that have not been backed up by sound, convincing research. After the introduction of the enamel acid-etch technique by Dr. Buonocore in 1955, the increasing demand for restorative and non-restorative esthetic treatments, as well as the refinement in the utilization of fluoride-containing products, have totally transformed the practice of operative dentistry. While bonding to enamel is a reliable technique, bonding to dentin represents a greater challenge: dentin is an intrinsically wet organic tissue penetrated by a tubular labyrinth containing the odontoblastic process, which communicates with the pulp. The manufacturers of new adhesive systems recommend the application of their adhesive materials on moist dentin. The main reason is that the spatial alteration that occurs upon drying demineralized dentin may prevent the monomers from penetrating the labyrinth of nano-channels formed by dissolution of hydroxyapatite crystals between collagen fibrils. Several of the current concepts in dentin bonding have been built from the observation of clinical studies, which measure retention rates, and from laboratory bond strength studies, which measure the force needed to debond a composite post from a flat dentin area. Although their utility is doubtful as far as clinical performance is concerned, bond strength studies are still necessary in order to compare similar parameters among materials. However, both clinical and laboratory studies lack information on the dynamic behavior of the substrate in a vital environment. For example, the changes induced by adhesive systems in the resin-infiltrated demineralized dentin have not been fully characterized. Also, the role of collagenous and non-collagenous proteins in dentin bonding has been generally overlooked. PMID- 11725669 TI - Durability of resin-dentin bonds. AB - PURPOSE: The purpose of this study was to determine if the durability of resin dentin bonds could be evaluated more quickly if the bond specimen was divided into 1 x 1 x 8 mm beams incubated at 37 degrees C for a 90-day period. MATERIALS AND METHODS: Extracted human third molars were prepared for bonding by removing the occlusal surface near the dento-enamel junction (superficial dentin group) or near the pulp (deep dentin group). The teeth were bonded either with MacBond, One Step or Clearfil Liner Bond 2, and then builtup to form a flat resin composite crown. After 24 hours in water, each buildup was vertically divided into slabs 1 mm thick, the top half of which was resin, with the bottom half as dentin. Each slab was then vertically sectioned at 1-mm increments to create 1 x 1 x 8-mm beams of resin-bonded dentin. They were incubated for 1 day or 90 days at 37 degrees C, followed by measurement of the tensile bond strengths. The results were analyzed by the Least-Squares Means method at the 95% confidence level. RESULTS: MacBond gave the highest (p < 0.05) 1-day bond strengths to superficial dentin, but significantly lower bond strengths were measured in deep dentin. There were no significant differences in the bond strengths of either One Step or Clearfil Liner Bond 2 to superficial vs deep dentin at 1 day, but at 90 days their bond strengths to deep dentin had fallen significantly (p < 0.05). Prepolymerized cylinders of resin composite bonded together with One Step showed little variation in bond strength over the 90-day experiment. SEM examination of the failed bonds showed increased porosity in intertubular dentin over time. CONCLUSION: The results indicate that division of large specimens into many small beams accelerated the deterioration of bond strength in deep dentin in all three bonding systems and in both superficial and deep dentin in the MacBond treated specimens. This method seems promising for studying the durability of resin dentin bonds. PMID- 11725670 TI - Influence of the time-point of salivary contamination on dentin shear bond strength of 3 dentin adhesive systems. AB - PURPOSE: The aim of this study was to evaluate the influence of the duration of salivary contamination at different stages during the bonding procedures on shear bond strengths (SBS) of 3 dentin adhesives. MATERIALS AND METHODS: Flat human dentin surfaces were produced by wet grinding on SiC paper 800. The adhesives used were Syntac Sprint (SS) (Vivadent), One Step (OS) (Bisco), Clearfil Liner Bond 2 (LB2) (Kuraray). Three durations of contamination (3 s, 10 s, and 20 s) at 3 stages (before adhesive application, after its application, and after its polymerization). In all cases the saliva was not rinsed off. Twenty-seven groups of 10 samples were studied according to 3 adhesives, 3 stages, and 3 durations of contamination. For each adhesive, 1 control group, 3 groups with contamination before adhesive application, 3 groups with salivary contamination on the uncured adhesive, and 3 groups with contamination after polymerization (except for SS) were studied. Composite cylinders (Z100, 3M; 3 mm O, 5 mm high) were polymerized on the surfaces and stored in 37 degrees C H2O for 48 hours prior to shear bond testing (v = 5 mm/min). RESULTS: Anova and Scheffe tests showed that for the one bottle systems tested, salivary contamination prior to adhesive application had no adverse effect on bonding efficacy, SBS decreased significantly when saliva contamination occurred after adhesive application. The self-etching primer tested was more tolerant to salivary contamination, except when the salivary contamination occurred before the polymerization of the adhesive. CONCLUSIONS: Salivary contamination does not have the same influence at different stages of the bonding process with modern adhesives. Pending better knowledge of the mechanisms involved in the influence of salivary contamination on bonded assemblies, it seems necessary to continue to recommend using the rubber dam in adhesive dentistry. PMID- 11725671 TI - In vitro leakage of resin-bonded all-porcelain crowns. AB - PURPOSE: The purpose of this study was to evaluate the sealing ability of single unit all-porcelain crowns with margins placed in cementum-dentin and luted with two different cementing materials. MATERIALS AND METHODS: Twenty extracted molars, free of caries and restorations, were selected. The sample abutments were prepared in a standardized manner and then divided in two groups (n = 10) at random. Group 1: The cervical margin was located 1 mm below the CEJ and the crowns were luted with EBS Multi (Espe) bonding system in combination with Compolute 1.0 Aplicap (CLA 1.0, Espe) experimental resin cement. Group 2: The cervical margin was placed 1 mm below the CEJ, and Syntac bonding system (Vivadent) and Variolink II resin cement (Vivadent) were used. IPS-Empress (Ivoclar) crowns were made following manufacturer's instructions. After luting procedures and 2,500 thermal cycles (5 degrees and 55 degrees C), the samples were processed and evaluated for marginal leakage at the cervical site. After scoring dye penetration in group 1 and 2 samples, half of the sections were randomly selected in order to evaluate 1) resin cement thickness and 2) hybrid layer formation at the interface between adhesive material and dental substrate. The leakage data were statistically evaluated with the Kruskal-Wallis non parametric analysis of variance (ANOVA) by ranks with Bonferroni alpha adjustment (p < 0.05). RESULTS: Sixty percent of the group 1 samples and 80% of group 2 showed leakage. In group 1, the dye penetration was often limited to the cervical margin site, while in group 2, it mainly extended to the axial walls. Statistically significant differences between samples of the two groups were not found. Samples of group 1 showed a cement thickness of 82 microns, while 75 microns was found in group 2. A hybrid layer and resin tag formation were evident in group 1. In group 2 samples, the hybrid layer formation was less evident and thinner than in group 1. CONCLUSIONS: The two adhesive systems performed similarly. The cervical margins of single-unit all-porcelain crowns must be considered as one of the weakest areas of this type of esthetic restoration. PMID- 11725672 TI - Shear bond strengths attained in vitro with light-cured glass ionomers vs composite adhesives in bonding ceramic brackets to metal or porcelain. AB - PURPOSE: The aim of this in vitro study was to evaluate whether light-cured resin modified glass ionomers provide sufficient bond strength for bonding ceramic brackets to metal and porcelain. In addition, the effect of mechanically retentive vs. silanized bracket bases on shear bond strength was investigated. MATERIALS AND METHODS: Flat specimens made from amalgam, a precious metal alloy, or ceramic were air-abraded with Al2O3 for 3 s and subsequently bonded with either Transcend 6000 or Fascination ceramic brackets. Alternatively, 20 porcelain specimens were treated with 9.6% HF for 2 min. Fuji Ortho LC and Photac Bond (light-cured glass ionomer cements [GIC]) served as adhesives. Two composite adhesives (Concise orthodontic and Transbond) in combination with various conditioners formed the control groups. After water storage for 24 hours, the shear bond strengths of all specimens were determined. RESULTS: Taking 10 MPa as a clinically sufficient bond strength, both resin-modified GICs produced a reliable retention of Transcend 6000 brackets on ceramic and amalgam, while the silanized Fascination brackets yielded a lower shear bond strength. On the precious metal alloy, only Fuji Ortho LC showed adequate strength. Various composite-conditioner combinations produced higher maximum bond strengths than the GICs. The location of bond failure varied considerably, and with most adhesives, bracket fractures occurred occasionally under shear stress. CONCLUSION: The investigated light-cured GICs provide sufficient strength for bonding ceramic brackets, but in terms of bond failure site and bracket fracture, they provide no advantage over composite adhesives. PMID- 11725673 TI - Clinical evaluation of adhesively placed Cerec endo-crowns after 2 years- preliminary results. AB - PURPOSE: Non-vital endodontically treated posterior teeth with complete loss of coronal hard tissues were prepared with a circular equigingival butt margin and central retention cavity of the entire pulp chamber ("endo-preparation"). Computer-generated ceramic corono-radicular restorations (Cerec endo-crowns) were bonded to these preparations. The purpose of this study was to evaluate the survival rate and the clinical quality of CAD/CIM endo-crowns after 2 years. MATERIALS AND METHODS: 19 Cerec endo-crowns (4 premolars and 15 molars) in 13 patients were examined using modified USPHS criteria at baseline and after an average time of 26 months. The ratings of the two examinations were compared. RESULTS: The service time of the 19 endo-crowns was 14 to 35.5 (mean +/- SD: 26 +/- 6) months. One molar endo-crown failed after 28 months because of recurrent caries. CONCLUSION: The overall clinical quality of the Cerec endo-crowns was very good, and so far, the clinical concept appears feasible. PMID- 11725675 TI - Adhesion: the silent revolution. 2nd European Symposium on Adhesive Dentistry: 7 9 May 1999. PMID- 11725674 TI - Minimally invasive restorative treatment. AB - Minimally invasive restorative treatment implements need-, damage-, and risk balanced dental care while ensuring that the ends and means are reconciled. Using examples from initial treatment of carious lesions, maintenance and correction of existing restorations, treatment of traumatic damage, and direct color and form correction of teeth, a preventively offensive and restoratively defensive treatment concept is presented. Such a concept demands of the dentist greater diagnostic efforts and more highly differentiated employment of invasive treatment methods. Demands on the patients are greater as well: they must be willing to comply with regular post-treatment examinations. In return, one receives the option of minimizing the sacrifice of maintainable hard tooth structures and avoiding extensive, large-scale dental procedures. PMID- 11725676 TI - Distribution of nanofillers from a simplified-step adhesive in acid-conditioned dentin. AB - PURPOSE: This in vitro study examined the interfacial ultrastructure of a nanofilled, simplified-step adhesive (Prime & Bond NT, Dentsply), to determine the distribution of nanofillers within the collagen network of the hybrid layer. MATERIALS AND METHODS: Twenty-four dentin discs were divided into two groups and bonded using two recommended conditioning techniques: Group I, NRC (Non-Rinse Conditioner), and Group II, Conditioner 36 (colloidal silica thickened 36% phosphoric acid). Following conditioning, a single coat of adhesive was applied and light-cured. Dentin discs were then bonded to form disc-pairs and processed for TEM examination. Demineralized, ultrathin sections were examined stained or unstained. Non-demineralized sections were used for STEM/EDX analysis of elemental distribution across the resin-dentin interface. In addition, four dentin discs were bonded with a generic adhesive (HEMA/TBBO) for TEM examination of stained collagen and proteoglycans. RESULTS: In unstained sections of both groups, nanofillers from the adhesive layer were congested around patent tubular orifices, but were not found within the interfibrillar spaces of the hybrid layer. EDX analysis of silicon (Si) showed predominant distribution within the adhesive layer and tubular orifices. Phosphorus (P) was present within the hybrid layer and adhesive layer in Group II. CONCLUSION: It is hypothesized that a) aggregation of the nanofillers within the adhesive resulted in filler clusters that are too large to infiltrate the interfibrillar spaces of the hybrid layer; and b) retention of ground substance within the demineralized intertubular collagen matrix may also have prevented the infiltration of the nanofillers. PMID- 11725677 TI - Variable polymerisation shrinkage and the interfacial micropermeability of a dentin bonding system. AB - PURPOSE: Adhesive bonds are subjected to competing stresses whilst a restoration is setting. This study aimed to determine the effect of these stresses on a chemically- and light-activated dentin adhesive (Scotch Bond MultiPurpose Plus: SBMP+) by assessing the morphology and micropermeability of the interfacial region. MATERIALS AND METHODS: Restorative materials and techniques were selected to impart differing setting stresses to the adhesive interface. An amalgam (Dispersalloy), a light-activated composite (Z100) with a high- or low-intensity curing light, and chemically-activated composite (Adaptic) were placed either in bulk (4 mm thickness) or in one thin layer (2 mm) in occlusal cavities in 55 freshly extracted third molars. In addition, thin composite restorations were placed on 15 horizontally sectioned teeth using a glass slide as a rigid clear matrix. The pulp chambers of all samples were filled with rhodamine B dye solution for 4 hours. Samples were longitudinally sectioned and the interfacial dye leakage measured using a confocal optical microscope. Data were analysed using the Kolmogorov-Smirnov test. RESULTS: In occlusal cavities, Z100 had significantly more leakage than the amalgam and Adaptic. There was no significant difference in micropermeability between large and small increments, or curing light intensity for Z100, but when placed in bulk, there was evidence of incomplete composite polymerisation near the cavity floor, especially with the low intensity curing light. Cohesive failure of the deep resin composite and gap formation were a frequent finding with the high-intensity light curing. The least leakage for Z100 was achieved with a thin layer on flat dentin. In contrast, the chemically-activated composite performed well in bulk when the surface of the restoration was freely exposed, but badly when constrained by the glass matrix. CONCLUSION: Restoration setting stress is critical in the microscopic integrity of the adhesive bond to dentin. PMID- 11725678 TI - Interfacial micromorphology and tensile bond strength of dentin bonding systems after NaOCl treatment. AB - PURPOSE: The objective of this study was to test the hypothesis that NaOCl treatment of acid-etched dentin would not lower the tensile bond strength of adhesive resins, but would eliminate the hybrid layer. MATERIALS AND METHODS: One hundred twenty teeth and three different bonding agents were used for SEM- and CLSM-visualization of the dentin-composite interface and for bond strength measurements. After etching with phosphoric acid, 75 dentin samples were pretreated with 10% NaOCl. The other 75 samples were etched only with phosphoric acid and were kept as controls. Composite was bonded to the pretreated surfaces with a bonding resin. RESULTS: NaOCl-treated samples: hybrid layers were not visible, but resin tags and resin penetration through lateral branches of tubules were prominent. Layers of unfilled resin were not apparent for Gluma CPS and Prime & Bond 2.1. Hypochlorite treatment of acid-etched dentin resulted in reduced bond strengths for specimens bonded with Syntac and Gluma CPS, but such treatment increased bond strength for Prime & Bond 2.1 bonded specimens. CONCLUSION: The removal of the collagen layer with NaOCl can enhance or decrease bond strengths, depending on the bonding agent used. PMID- 11725679 TI - Marginal adaptation of compomer Class V restorations in vitro. AB - PURPOSE: The purpose of this in vitro study was to determine the marginal adaptation of different adhesives, including recently developed self-etching primers, in combination with compomers in Class V cavities. MATERIALS AND METHODS: In 80 extracted human teeth, divided into 10 groups of 8, Class V cavities were prepared (ca. 4 mm high, 3 mm wide, and 1.5 mm deep) with one half of the margin length in dentin. The one-bottle adhesives Prime & Bond 2.1, Syntac SC, and Espe OSB, and the self-etching primers Rasant F 2000 and Prompt L Pop were used in combination with compomers. Before and after thermocycling (TC), replicas were taken and a quantitative margin analysis using the SEM was performed at a magnification of 200x using defined criteria. RESULTS: The statistical analysis using Kruskal-Wallis and Bonferroni tests showed a significantly (p < 0.05) better marginal adaptation in enamel and dentin for the self-etching primers, and when the one-bottle adhesives were used in combination with the total-etch technique. CONCLUSION: Self-etching primers improve the marginal adaptation of compomer restorations in enamel. PMID- 11725680 TI - Intermittent loading of teeth restored using quartz fiber, carbon-quartz fiber, and zirconium dioxide ceramic root canal posts. AB - PURPOSE: The purpose of the investigation was to compare the performances of teeth restored with quartz fiber, carbon-quartz fiber, and zirconium-dioxide posts covered with all-ceramic crowns when subjected to a cyclic loading test performed in a wet environment. MATERIALS AND METHODS: Forty single-rooted human lower premolars having similar dimensions were endodontically treated and mounted in acrylic resin blocks with a simulated periodontal ligament. The teeth were divided into three experimental groups and one control group. Post holes 8 mm long were prepared in the roots of the experimental groups in which quartz fiber, carbon-quartz fiber, and zirconium dioxide posts were cemented. In the control group, no posts were used. The crown buildup was made with composite resin. The teeth were covered with all-ceramic crowns and intermittently loaded at an angle of 45 degrees to the long axis of the tooth at a frequency of two loads per second. RESULTS: Only one failure (root fracture + post fracture) was observed in each of the fiber post groups, while in the zirconium dioxide post group, six failures were observed (one crown fracture and 5 root fractures + post fractures). The Kaplan-Meier analysis of the three experimental groups showed that the survival rate of zirconium dioxide posts was significantly lower than that of both types of fiber post. All the experimental groups showed a survival rate higher than that of the control group. CONCLUSION: Fiber posts reduced to a minimum the risk of root fractures of teeth restored with composite cores and Empress crowns under the present experimental conditions (intermittent loading in a wet environment). PMID- 11725681 TI - The influence of a simplified application technique for ceramic inlays on the margin quality. AB - The adhesive luting technique for ceramic inlays requires a sequence of multiple steps that are cumber-some and time consuming. Furthermore, modifications of dentin bonding procedures may negatively influence the margin quality of ceramic inlays with cervical dentin finishing lines. PURPOSE: The objective of this study was to test the effect of a new, simplified procedure for dentin bonding on the margin quality of ceramic inlays. MATERIALS AND METHODS: Thirty-two MOD inlay cavities with cervical finishing lines in dentin were prepared into premolars. They were randomly divided into four groups (n = 8): In the balanced design, one parameter was the inlay material (Empress/Cerec) and the other the polymerization of the dentin bonding agent, before insertion of the inlay (P+) and after insertion of the inlay (P-). All inlays were inserted with Variolink II in combination with an enamel and dentin conditioning universal bonding agent (Etch & Prime 3.0). Two parameters were used to determine the inlay quality: (1) the fit, measured by the difference of the insertion depth into the cavity between try-in and cementation, and (2) a quantitative margin analysis in the SEM at a magnification of 200x, performed for the cervical segment before and after thermocycling, using epoxy replicas. RESULTS: In all four groups the inlays were > 5 microns deeper in the cavities than at the try-in session. This is explained by the substance loss from the etching. The measured difference for Cerec inlays inserted in cavities, where the bonding agent was not polymerized prior to insertion, was significantly larger. Margin analysis of the cervical segment revealed no statistical differences (P < 0.01) for both criteria tested. The median values for all groups were 100% for clinical acceptable margin and 0% for marginal opening. CONCLUSION: By using the universal bonding agent, high margin qualities on dentin and enamel are obtained with ceramic inlays even with the simplified approach. PMID- 11725682 TI - Direct posterior composite restorations with a new adhesive system: one-year results. AB - PURPOSE: Syntac Sprint is a new one-bottle adhesive based on the total-etch and total-bond technique. The aim of this study was to compare the new restorative system consisting of Syntac Sprint and the resin composite Tetric Ceram with the clinically proven combination of the dentin adhesive Syntac Classic and the resin composite Tetric. MATERIALS AND METHODS: In a controlled prospective clinical study, 47 cavities (36 premolars, 11 molars; 29 maxillary, 17 mandibular; 6 occlusal, 33 occluso-approximal, 7 MOD, 1 > 3 surfaces) in 33 patients (17 female, 16 male) were filled. Twenty-five fillings were placed with the combination Syntac Sprint/Tetric Ceram under total etching and total bonding conditions. In thirteen restorations, Syntac Sprint was precured prior to the application of the resin composite; in the other twelve cases, restoration and adhesive were cured simultaneously. The control group consisted of 22 cavities restored with Syntac Classic plus the composite Tetric. At baseline and after 6 and 12 months, two calibrated investigators examined the restorations with modified USPHS codes and criteria. RESULTS: The criteria marginal integrity and integrity of the tooth deteriorated significantly between the baseline and the 6- and 12-month investigations (Friedman 2-way ANOVA; p < 0.05). No differences were found in the criteria surface roughness, color matching, anatomic form, step formation, integrity of the filling, proximal contact, hypersensitivity, or satisfaction. After one year, all fillings were in good condition. There was no significant difference between the Syntac Sprint/Tetric Ceram and Syntac Classic/Tetric groups for any criterion (Kruskal-Wallis; p > 0.05). CONCLUSION: Within the limits of this study, Tetric Ceram fillings in combination with Syntac Sprint were clinically successful after one year. PMID- 11725683 TI - Tensile bond strength of composite to air-abraded enamel. AB - PURPOSE: The purpose of this study was to test the tensile bond strength of composite resin to enamel surfaces treated with kinetic cavity preparation (KCP) and acid etching, respectively. MATERIALS AND METHODS: Plane labial surfaces of 280 human maxillary central incisors were treated with either KCP by varying treatment conditions or phosphoric acid gel. Composite samples were bonded to the pretreated surfaces with a bonding resin. After 24 hours specimens were loaded in tension in an Instron testing machine until bond failure. A two-way analysis of variance was used to determine significant differences. The failure sites were qualitatively evaluated by SEM. RESULTS: The determined tensile bond strengths and observed failure modes demonstrated that acid-etched enamel has a significantly higher bond strength to composite resin than KCP-treated enamel surfaces. CONCLUSION: KCP-prepared enamel has to be acid etched before bonding to composite resin. PMID- 11725684 TI - Marginal adaptation of Class II composite fillings: guided polymerization vs reduced light intensity. AB - PURPOSE: To improve the marginal adaptation of Class II composite fillings, a three-sited light-curing technique was proposed by Lutz et al in 1986 and has proven to be effective in several studies. This study investigated whether this effect, attributed to guided polymerization towards the cavity margins, might also be due to the decreased polymerization stress resulting from reduced curing light intensity caused by transdental curing. MATERIALS AND METHODS: Forty extracted human molars were mounted in casts and 40 Class II slot-type cavities were cut (cervical margins 0.5 mm above the CEJ). To measure the light being transmitted from a halogen curing light onto a composite layer within a cavity, miniature photodetectors were used and irradiance measured for a three-layer technique (1st: cervical-horizontal; 2nd and 3rd: lateral-diagonal) with occlusal or three-sited curing. To evaluate the influence of light intensity on marginal adaptation, the prepared cavities were treated as follows: after base placement and acid etching, the cavities were filled with Herculite XR (Kerr, Romulus, MI, USA) in three increments. All cavities were filled with the layering technique described above. In two groups (n = 10), curing was carried out only from occlusal. In the remaining groups, the cervical composite layer was cured through a light-transmitting, lateral reflective wedge, and the following layers transdentally with a proximal wand positioning. Using the photodetectors, the output power of an argon laser was adjusted so that the composite was cured with the equivalent irradiance calculated for occlusal (high-intensity groups) and three-sited curing occlusal (low-intensity groups). Replicas were produced after storing in water for 21 days and thermocycling (2000x, 5 degrees C to 55 degrees C). The margins of the fillings were quantitatively assessed using a SEM at 200x. The results were statistically analyzed (ANOVA, Bonferroni/Dunn; p < 0.05). RESULTS: The summed values for the criteria "restoration margin fracture", "enamel margin fracture," and "marginal opening" at the cervical margins were 11.3 +/- 11.5 (three-sited curing, low intensity), 14.1 +/- 15.7 (occlusal curing, low intensity), 25.4 +/- 21.2 (three-sited curing, high intensity), and 27.8 +/- 30.6 (occlusal curing, high intensity). CONCLUSION: No significant difference was found between curing techniques at equal energy levels; the effect of the three-sited light curing technique should therefore not be attributed to guided polymerization but to the reduction of light intensity. PMID- 11725685 TI - Evaluation of the marginal and internal adaptation of different ceramic and composite inlay systems after an in vitro fatigue test. AB - PURPOSE: The aim of this in vitro trial was to evaluate and compare the external and internal adaptation of Class II composite and ceramic inlays with scanning electron microscopy after mechanical loading and thermal cycling (fatigue). MATERIALS AND METHODS: Standardized cavities were prepared on human third molars with margins extending proximally 1 mm above or below the cemento-enamel junction (CEJ). A fine hybrid composite (Tetric), a hydrothermal glass (Ducera LFC), and a sintered spinell ceramic (In-Ceram) were used to fabricate the restorations. Combinations of Syntac classic--Variolink, ED primer--Panavia and Scotchbond MP- Variolink were used for cementation. RESULTS: For marginal adaptation in enamel, percentages of continuity varied from 85.4% to 99.6% and 85.1% to 98.2% for mesial and distal sites, respectively. Marginal adaptation in dentin showed continuity percentages varying from 63.6% (In-Ceram-Panavia) to 96.1% (Ducera LFC Panavia). As regards internal adaptation, overall interface results varied from 32.1% (Tetric-Variolink) to 87.3% (Ducera LFC-Panavia). Gingival enamel showed the higher proportions of continuity (94.0%, all group-pooled data) compared to dentin locations (46.0% occlusally to 60.1% in gingival dentin). In all groups, debonding occurred mainly within the dentin--restoration interface and also, but to a lesser extent, between In-Ceram ceramic and both luting composites. When it occurred, adhesive failure to dentin consistently took place at or close to the hybrid layer surface. CONCLUSION: Despite important differences in their physical properties, no difference could be found under the present experimental conditions between Class II composite and ceramic inlays in terms of their marginal and internal adaptation. PMID- 11725686 TI - Do dentin bond strength tests serve a useful purpose? AB - PURPOSE: The publication of dentin bond strength data is prolific in the dental literature and will remain so as more and more new dentin bonding agents are marketed. It is the purpose of this presentation to assess the usefulness of this information. METHODS: The limitations of dentin bond strength tests are examined in the context of current knowledge about the methods used. RESULTS: The evidence provided shows that dentin bond strength measurement techniques provide data that is not able to act as a consistent predictor of clinical performance. Fundamental flaws in the most popular methods adopted have been identified and alternative approaches need to be explored. Unfortunately, we do not as yet have in place methodologies that allow us to establish a reliable link between laboratory bond strength data and clinical performance. CONCLUSION: For the present, clinically based evidence remains the only reliable means for the selection of dentin bonding agents. PMID- 11725687 TI - Hybridization effectiveness of a two-step versus a three-step smear layer removing adhesive system examined correlatively by TEM and AFM. AB - PURPOSE: The objectives of this study were (1) to compare the hybridization effectiveness of two adhesive systems that are applied in respectively three and two steps, and (2) to determine the best resin-dentin interface preparation technique for atomic force microscopy (AFM). MATERIALS AND METHODS: The resin dentin interface produced by the three-step OptiBond Dual-Cure (Kerr) and its simplified two-step successor OptiBond Solo (Kerr) was ultramorphologically examined using transmission electron microscopy (TEM) and AFM. Four different methods were used to prepare interface specimens for AFM: (1) polishing to a 0.1 micron finish with a silicon oxide suspension, (2) polishing to a 0.05-micron finish with an aluminum oxide suspension, (3) argon-ion etching, and (4) sectioning with a diamond knife. RESULTS: Both TEM and AFM demonstrated that some collapse of the exposed collagen fibril network, due to gentle postconditioning air-drying of the dentin surface, may not have been totally recovered through hybridization by the two-step adhesive formulation as opposed to the three-step precursor. From the four interface preparation methods, only diamond-knife sectioning revealed sufficient ultramorphologic detail and high resolution that can capitalize on the high resolution offered by AFM. CONCLUSION: First, the findings suggest that simplifying the application procedure of adhesives by combining the primer and adhesive resin into a single application step may reduce hybridization effectiveness. Future research should confirm this effect for other two- versus three-step adhesive systems. Second, diamond-knife sectioning should be used for future topographic imaging and physicomechanical testing of resin dentin interfaces by AFM. PMID- 11725688 TI - Ceramic inlays: clinical assessment and survival rate. AB - PURPOSE: This study evaluates the clinical behavior of ceramic inlays placed during the past decade. MATERIALS AND METHODS: One hundred eighty-two inlays were examined in 66 patients. The interval between placement and assessment was on average 5.9 years +/- 2.7, ranging from 2 to 11.7 years. Restorations still present at the time of evaluation were clinically assessed according to modified USPHS criteria. Kaplan-Meier statistical analysis was used to assess the survival rate. RESULTS: According to USPHS criteria, good results were obtained for color match, marginal discoloration, recurrent caries, contour, and marginal integrity. Six inlays failed: four for endodontic reasons, one due to recurrent caries, and the other due to fracture. The results indicate that a success rate of 95% could be predicted at 11.5 years. CONCLUSION: The lack of recurrent caries, the only slight changes in marginal discoloration and color match, combined with the excellent longevity prove that ceramic inlays are a valuable tool for the restoration of posterior teeth. PMID- 11725689 TI - Additive contour of porcelain veneers: a key element in enamel preservation, adhesion, and esthetics for aging dentition. AB - Esthetics and function are equal concerns when restoring the anterior dentition. Modern concepts in restorative dentistry have brought new solutions through bonded porcelain veneers that are stress distributors and involve the crown of the tooth as a whole in supporting occlusal force and masticatory function. This recovery of the original biomechanics of the intact tooth, the biomimetic principle, is particularly valuable when considering the restoration of an aging dentition. Both function and appearance are affected by the senescent changes of the aging teeth. Erosion and surface wear lead to a progressive thinning of enamel, ultimately leading to increased crown flexibility and higher surface strains. It appears therefore that the restoration of tooth volume will not only re-establish the original and youthful appearance of the smile but will also allow the biomimetic recovery of the crown. The final treatment outcome strongly depends on the therapeutic approach chosen, the driving force of which should be the preservation of the thin remaining enamel. While a number of preparation techniques will expose dentin to a great extent, the principle of enamel preservation can still be fulfilled by the use of a specific approach. This article describes a treatment method which includes the use of a diagnostic template. This type of work strategy, documented with clinical cases, integrates additive wax-ups and acrylic mock-ups. The latter will provide a significant amount of diagnostic information and economy of tooth substrate, the importance of which cannot be overestimated in the completion, functionality, and longevity of the final restoration. PMID- 11725690 TI - Effect of alpha-tocopherol on hepatocarcinogenesis in transforming growth factor alpha (TGF-alpha) transgenic mice treated with diethylnitrosamine. AB - To examine the potentially chemopreventive effects of alpha-tocopherol on hepatocarcinogenesis, we fed the transgenic mice line MT42, which overexpresses transforming growth factor-alpha (TGF-alpha) and which has been established as having a high incidence of liver tumor, with different concentrations of alpha tocopherol and examined the hepatic tumorigenesis of these mice. At 3 weeks of age, MT42 male mice received a single intraperitoneal injection of diethylnitrosamine (DEN), 5 mg/kg body weight, to initiate the formation of liver tumors. The mice were divided into three groups: group A, control diet (20 mg/kg of alpha-tocopherylacetate); group B, deficient diet (less than 1 mg/kg); group C, supplemented diet (500 mg/kg). Neoplastic change was determined at 40 weeks of age. The incidence of adenomas (p < 0.05), the maximum tumor size (p < 0.01), the mean relative liver weight (p < 0.01), and the proliferating cell nuclear antigen (PCNA) labeling indices of the non-tumor sites (p < 0.01) of group B were significantly higher than those of group C. No toxic effects of alpha-tocopherol were found. Alpha-tocopherol-deficient diet accelerated the hepatocarcinogenesis of TGF-alpha transgenic mice treated with DEN. At best, these data demonstrate that alpha-tocopherol-deficiency is not beneficial for prevention of hepatocarcinogenesis in this model. Alpha-tocopherol may be useful for the chemoprevention for liver cancer. PMID- 11725691 TI - A national survey of iron and folate status in pregnant women in Switzerland. AB - Women often do not meet the increased iron and folate needs of pregnancy. Maternal iron-deficiency anemia is associated with poor maternal and infant outcomes, including preterm delivery and low birth weight. Poor folate status increases risk for maternal anemia, spontaneous abortion, and congenital defects. Because of this, supplemental iron and folate are often recommended during pregnancy. There are few data on iron and folate status in pregnant women in Switzerland. We measured iron and folate status in a national sample of Swiss pregnant women, estimated the prevalence of anemia, and determined if supplement use is associated with iron and/or folate status in this group. A 3-stage probability to size cluster sampling method was used to obtain a representative national sample of pregnant women (n = 381) in the second and third trimester. We measured hemoglobin, hematocrit, mean corpuscular volume, and serum folate and ferritin concentrations. Serum transferrin receptor concentration was determined in anemic subjects. The use of iron and folate supplements was evaluated by questionnaire. Mean hemoglobin (+/- SD) in the sample was 123 g/L (+/- 1.0). The prevalence of anemia was 6%. Of the 21 anemic women, 11 were iron-deficient, giving an iron-deficiency anemia prevalence of 3%. Nineteen percent of women had low serum ferritin concentrations (< 12 micrograms/L) and 4% had low serum folate concentration (< 2.5 micrograms/L). Supplements containing iron were taken by 65% of women, and 63% were taking folate-containing supplements. Women in the second and third trimester taking folate-containing supplements had significantly higher serum folate concentrations compared to those not taking a folate supplement (p < 0.001). In the third trimester, women taking iron-containing supplements had significantly higher serum ferritin concentrations compared to those not taking an iron-containing supplement (p < 0.01). Our findings indicate that iron and folate status appears to be adequate in the majority of pregnant women in Switzerland, and that use of iron and folate supplements may have a positive impact on status. PMID- 11725692 TI - Nutritional and food protection against epidemic emerging neuropathy. Epidemiological findings in the unique disease-free urban area of Cuba. AB - A survey was conducted through the SECUBA (SEguridad alimentaria en CUba y Buena Alimentacion) research program in Cuban healthy smokers living in Guantanamo and in Havana. The aim of the survey was to investigate biological and nutritional factors connected with the occurrence of zero epidemic neuropathy (EN) observed in Guantanamo urban area since the disease emerged in Cuba. Blood riboflavin status and carotenoid and selenium concentrations were higher in Guantanamo than in Havana smokers. Food dietary quantities of plantain banana, pepper (Capsicum spp.), bovine meat and milk products were higher in Guantanamo. Inversely, foods rich in cholesterol, especially eggs, were more consumed in Havana. Through riboflavin, carotenoid and selenium contents and specific antioxidants substances (indoleamines, capsaicin), the foods more consumed in Guantanamo could be considered as EN protective factors. Disease protective effects could be exerted via enhancement of defence mechanisms against free radical damage and related mechanisms focused on redox recycling of glutathione and local protection from carotenoids. Finally, the results of the present study should help Cuba, through a better EN control, to improve long-term food safety and define healthier dietary habits. PMID- 11725693 TI - Maternal-fetal n-6 and n-3 polyunsaturated fatty acids gradient in plasma and red cell phospholipids. AB - Fatty acid distribution was investigated in ethnically and economically homogenous Korean mothers (n = 40) and neonates. Venous blood, maternal before delivery and cord, was obtained. Choline (CPG) and ethanolamine (EPG) phosphoglycerides and sphingomyelin (SM) were assayed. Mean arachidonic acid (AA) level was higher in plasma CPG and SM (p < 0.0001), and red cell CPG (p < 0.0001), EPG (p < 0.0001) and SM (p = 0.005) of the neonates. Similarly, the neonates had higher proportions of docosahexaenoic acid (DHA) in plasma CPG (p < 0.0001) and red cell CPG (p = 0.001) and EPG (p = 0.036). In contrast, linoleic and alpha-linolenic acids were significantly higher in maternal blood. Mead acid was elevated in plasma CPG (p < 0.0001) and red cell CPG and EPG (p < 0.0001) of the neonates. Consistent with data from high-fat-intake populations, our subjects, whose traditional diet is low in fat, exhibited maternal-fetal gradient in AA and DHA in plasma and red cell phospholipids. This may be due to an imbalance between supply and maternal and fetal requirements, and/or a physiological response to pregnancy. Prenatal nutritional constraint is associated with impaired development and a risk of chronic diseases in adults. AA and DHA are vital nutrients. Hence, there is a need to investigate whether the discrepancy between maternal and neonatal AA and DHA is a manifestation of nutritional insufficiency. PMID- 11725694 TI - Effects of androstenedione-herbal supplementation on serum sex hormone concentrations in 30- to 59-year-old men. AB - The effectiveness of a nutritional supplement designed to enhance serum testosterone concentrations and prevent the formation of dihydrotestosterone and estrogens from the ingested androgens was investigated in healthy 30- to 59-year old men. Subjects were randomly assigned to consume DION (300 mg androstenedione, 150 mg dehydroepiandrosterone, 540 mg saw palmetto, 300 mg indole-3-carbinol, 625 mg chrysin, and 750 mg Tribulus terrestris per day; n = 28) or placebo (n = 27) for 28 days. Serum free testosterone, total testosterone, androstenedione, dihydrotestosterone, estradiol, prostate-specific antigen (PSA), and lipid concentrations were measured before and throughout the 4-week supplementation period. Serum concentrations of total testosterone and PSA were unchanged by supplementation. DION increased (p < 0.05) serum androstenedione (342%), free testosterone (38%), dihydrotestosterone (71%), and estradiol (103%) concentrations. Serum HDL-C concentrations were reduced by 5.0 mg/dL in DION (p < 0.05). Increases in serum free testosterone (r2 = 0.01), androstenedione (r2 = 0.01), dihydrotestosterone (r2 = 0.03), or estradiol (r2 = 0.07) concentrations in DION were not related to age. While the ingestion of androstenedione combined with herbal products increased serum free testosterone concentrations in older men, these herbal products did not prevent the conversion of ingested androstenedione to estradiol and dihydrotestosterone. PMID- 11725695 TI - Vitamin D insufficiency in Japanese female college students: a preliminary report. AB - Vitamin D insufficiency, a risk factor for osteoporosis, has been well investigated in elderly women worldwide, but little information has been available for younger women. This study aimed to determine serum 25 hydroxyvitamin D [25(OH)D] levels in Japanese female college students, and to test whether there is an association between 25(OH)D and bone strength. Subjects were 77 Japanese female junior college students aged 19-24 years. Serum 25(OH)D was measured with high performance liquid chromatography. Bone strength of the calcaneus was evaluated using the osteo sono-assessment index (OSI), a combined index reflecting the bone mass, bone architecture, and elasticity, as measured by quantitative ultrasound densitometry (AOS-100, Aloka). The mean 25(OH)D concentration was 34.2 nmol/L (SD +/- 12.1). The proportion of subjects with 25(OH)D less than 30 nmol/L, a cut-off value for vitamin D insufficiency, was 31/77 (40.3%). Simple linear regression analysis showed that there was a significant linear relationship between 25(OH)D and OSI (R2 = 0.098, p = 0.0069). The association held even after adjusting for weight (partial R2 = 0.098, p = 0.0023). Body weight was also a significant predictor of OSI (partial R2 = 0.105, p = 0.0034). These results suggest that vitamin D insufficiency may be prevalent in healthy young women. Moreover, low levels of 25(OH)D in young women may adversely affect bone strength. An additional, newly designed epidemiological study with a sufficient sample size is needed to confirm the present findings. PMID- 11725696 TI - Acute and chronic toxicity study of fish oil and garlic combination. AB - The purpose of this study is to evaluate the acute and chronic toxicology of oral intake of fish oil (omega-3 fatty acid) and garlic combination food supplements. These supplements were proven to have beneficial effects on the lipid profile. Therefore, it is important to evaluate the potential long-term effects of fish oil and garlic combination supplements on the biochemistry of organ structure and function. The hypothesis to be tested was that acute and chronic high-dose supplements of fish oil and garlic may not adversely affect organ histology but may influence certain metabolic activities. A double-blind, placebo-controlled study was carried out using 28 Sprague Dawley rats separated into a placebo group (16 rats) and a supplement group (12 rats). The supplement group received the ingredients in chow inserts at a dosage that was equivalent to three times the maximum safe daily dosage for fish oil and the usual daily dosage for garlic (the maximum safe daily dosage recommended by the United States Food And Drug Administration for a 70-kg human is a total of 3 g/day intake of EPA and HDA omega-3 fatty acids from conventional and dietary sources. The usual daily garlic usage is garlic powder = 1200 mg). The study was conducted over a period of 12 months with evaluations performed at baseline, 2 months, 6 months, and 12 months. Results confirm the expected acute triglyceride, total cholesterol and LDL suppression at these higher dosages in the supplement group. Acutely and chronically, there were no differences in external appearance, level of activity, daily food consumption, blood cell count, kidney function, thyroid function, prothrombin time (PT), and activated partial prothrombin time (PTT), which remained within normal ranges in the supplement group. Organ histology remained unchanged. Although during the chronic toxicity period the triglyceride and LDL suppression persisted, it was noted that total cholesterol and HDL levels increased. The increase in cholesterol and HDL in the supplement group during chronic toxicity periods is simultaneous with loss of suppression of plasma levels of other liver function marker enzymes, ALT and AST, which are not involved in cholesterol synthesis. This possibly suggests that other liver enzymes involved in cholesterol synthesis, such as HMG-co A reductase, follow a similar escape from suppression. PMID- 11725697 TI - Vitamin B6 intakes and status assessment of elderly men and women in Taiwan. AB - Studies have indicated that vitamin B6 status decreases with age. However, little is known about vitamin B6 status of elderly people in Taiwan. The purpose of this study was to assess vitamin B6 status of elderly Taiwanese and to examine the effect of protein on various indices of vitamin B6 status in the elderly. Thirty nine men (mean age = 69.9 +/- 4.2 years) and 55 women (mean age = 69.5 +/- 3.9 years) completed a 24-h diet recall. The mean total vitamin B6 intake (men: 1.7 +/- 0.9 g/day; women: 1.6 +/- 1.2 g/day) was higher than the 1998 US Dietary Reference Intakes (DRI) and the current Taiwan Recommended Daily Nutrient Allowance (RDNA). Dietary energy and protein intakes were not related to any vitamin B6 status parameters in any sex groups and the pooled group. Vitamin B6 intake correlated only with erythrocyte aspartate transaminase activity coefficient (EAST-AC) in the pooled (r = -0.214, p < 0.05) group. There were no significant differences in plasma pyridoxal 5'-phosphate (PLP), erythrocyte alanine transaminase activity coefficient (EALT-AC), and EAST-AC between sex groups. Although elderly subjects had adequate mean plasma PLP concentrations, 59% of men and 55% of women had plasma PLP concentrations lower than a cutoff of 20 nmol/L. The mean EALT-AC was < 1.25 in two groups with adequate vitamin B6 status. However, 23% of men and 18% of women had EALT-AC values > 1.25. The mean EAST-AC value of subjects was higher than the suggested value (< 1.8) for inadequate vitamin B6 status. The incidence of biochemical vitamin B6 deficiency in our elderly is probably more relevant from other causes than from dietary intake of protein and vitamin B6. PMID- 11725698 TI - [Africa or Asia, which is the evolutionary origin of human schistosomes?]. AB - The origin and the evolution of Schistosomatidae species, due to their medical importance (responsible of the second most important human parasitosis after malaria), arouse a great interest. A combination of phylogenetic studies using several molecular markers has provided support for the traditional grouping and evolutionary inferences derived from morphological and biological data. The genus Schistosoma, which comprises all species parasitizing Man, is generally split into four evolutionary lineages (mansoni, haematobium, indicum and japonicum lineages). The group of African schistosomes (including mansoni and haematobium lineages) appears very divergent from the japonicum lineage. Recent phylogenetic studies using partial 28S rDNA sequencing and including Orientobilharzia turkestanicum from Iran, an Asian parasite of livestock, found, unexpectedly, that this species nested among Schistosoma species, thus rendering the latter paraphyletic, and suggested an Asian origin for the Schistosoma genus. The present work re-examines the question of the geographical origin of human schistosomes by analysing a new genomic marker (ITS2) as well as by including the use of O. turkestanicum originating from northeastern China. Our results are in agreement with previous work using 28S, in demonstrating that Schistosoma is not monophyletic. However, O. turkestanicum, whatever the method of analysis used (distance or parsimony), was grouped with members of the japonicum group to the exclusion of African Schistosoma species. Then, our data argue strongly for the need for further phylogenetic study including new taxa and new genomic sequences before definitely concluding either an Asian or African origin for the genus Schistosoma. PMID- 11725699 TI - [Larval history of glass eels, Anguilla anguilla (Linne, 1758) in migration in coastal and estuary areas (Adour, Gulf of Gascogne) as revealed by otolith test]. AB - The embryonic past of glass eels was studied from the interpretation of microstructures registered on otoliths. The aim of this work is to put in evidence possible seasonal modifications of the growth of otoliths so that differences between otoliths of glass eels caught off marine and estuarine environment. So during the season 1999-2000, from November till March, otolith sampling was realised in the southwestern part of France, in an estuarine and coastal zone. We observed a spatial and temporal evolution of proportions of the three various types of otoliths taken into account. Glass eels sampled at sea sometimes have a mark on their otoliths indicating the transition in the estuary, especially at the end of the fishing season. Measures of growth marks of otoliths showed that there were no seasonal differences during phases of the transoceanic migration and the crossing of the continental shelf. The radius of otoliths of glass eels sampled at sea was significantly smaller than those sampled in estuary. These results translated homogeneous environmental modifications met by the various larvae groups during the oceanic crossing and during the principal migration season as well as a turn over of these groups during the transition between marine and continental environment. PMID- 11725700 TI - Experience modulates emission of food calls in broody hens. AB - The aim of these experiments was to determine if previous experience of chicks' response to food calling influences subsequent propension of maternal hens to utter food calls. Seventeen broody hens were tested three times a day without their 3- or 4-day-old chicks. Hens were tested in two situations: chicks were returned either after each test or at the end of all the day's tests. As palatability influences food calling in maternal hens, experiments were conducted first with a highly preferred food item and then with the hens' usual feed. The chicks' capacity to respond regularly to their mother influences the hens' capacity to emit food calls. In fact, although the hens did not lose their maternal state, they uttered fewer food calls when their chicks were removed all day. These results suggest that the chicks' behaviour following food calling could be a social reinforcement for broody hens. PMID- 11725701 TI - Studying boundary effects on animal movement in heterogeneous landscapes: the case of Abax ater (Coleoptera: Carabidae) in hedgerow network landscapes. AB - Changes in movement possibilities of insect species can be a response to increasing heterogeneity in man made landscapes. We studied the movement of Abax ater (Coleoptera, Carabidae) between woody and cultivated areas in Brittany. Two woody habitats (woods and hedgerows) and two cultivated areas (cornfields and meadows) were considered. Quantifying the movement of the carabid beetle between woody and cultivated areas was done using Mark-Release-Recapture techniques. Marked individuals were released at the centre of circular enclosures (3 m in diameter), encompassing two adjacent habitats, one being wooded, the other cultivated. Individuals were recaptured at the edge of enclosures. Analysis of recaptures showed that flows of individuals from a given habitat to another were constant during the experiment. Flows can be summed up by a probability of move, which depends on the two adjacent habitats surrounding ecotone. When released in cultivated habitats, forest carabid beetles preferentially moved toward woody habitat, but in some enclosures the forest carabid beetles did not exhibited any preference between cultivated areas and woods. In these cases ecotones may include both woody and cultivated areas, and their functional width should be reconsidered. These conclusions were globally in agreement with knowledge about ecology of Abax ater, but ask for confirmation through replicates on different sites. Nevertheless, these results demonstrated the usefulness of the method and give new guidelines for the study of flows of carabid beetles through ecotones. PMID- 11725702 TI - [Nyctohemeral variations of marine bioluminescence in the Mediterranean and the northeastern Atlantic]. AB - Bioluminescence measurements have been made using a bathyphotometer allowing the determination of stimulated light intensities down to 2,000 m depth, in the Mediterranean Sea on the Almeria-Oran front, during the winter 1997-1998, and in the northeastern Atlantic, on the Armorican continental shelf, during summers 1999 and 2000. Bioluminescence is weaker in the Mediterranean than in the Atlantic. In the epipelagic waters, day/night variations appear clearly, stimulated bioluminescence is higher at night than during the day. These diel variations can be explained by vertical migration of bioluminescent organisms and by photoinhibition of dinoflagellate bioluminescence. Fluorescence measurements made at the same time give information about potential bioluminescent sources, autotrophic and heterotrophic. PMID- 11725703 TI - Identifying main evolutionary mechanisms shaping genetic variation of Leuciscus cephalus L. 1758 (Cyprinidae) in western Greece: discordance between methods. AB - Numerous methods can be used in intraspecific phylogeographic studies to infer the evolutionary mechanisms that shaped observed genetic variation in populations. However, these methods are scarcely used jointly, and the evolutionary outcomes they could propose are not fully compared. In this study, using a chub (Leuciscus cephalus; Cyprinidae) mitochondrial DNA data set (13 populations in Western Greece, 14 haplotypes), we compare three distinct 'historical' methods that could possibly infer relative importance of basic evolutionary mechanisms (isolation vs migration) shaping genetic variation: the nested clade analysis, the psi-test and the 'mismatch distributions'. Taking together, interpretations of these analyses allow to draw a picture of the evolutionary history of chub in Western Greece based on isolation and genetic drift for higher clades. However, results issued each method can differ for low differentiated clades. We discuss such differences and suggest that methods should be used jointly in phylogeographic studies for a better evaluation of the evolutionary mechanisms that shaped genetic variation. PMID- 11725704 TI - [Distinctive morphofunctional features of the bill and hyoid apparatus of turacos (Aves, Musophagidae): relations with frugivory]. AB - The present study points to morpho-anatomical features that allow vegetarian but mainly frugivorous turacos to fill at best their specific feeding requirements. Mechanisms are analysed which the bird can use to detach a fruit and avoid it rolls out of the bill. It is also examined how vegetable items can be processed particularly when they have to be cut, and what can facilitate their ingestion particularly when they are fruits that have to be swallowed entirely and are large and/or have dry or fibrous skin or flesh. The skeletal and muscular anatomy of the bill and hyoid apparatus is described in details and illustrated. The particularly short and stout bill coupled with a relatively simple system of aponevroses of the adductors reflects a trade-off between two important jaw functional requirements: huge efforts for clamping, and a wide opening for plucking, processing and swallowing relatively large fruit. The clearly oblique orientation of the modified outer adductor seems an adaptation to the detachment of fruits. The os uncinatum, particularly developed in this bird family, is interpreted as maintaining transversal stability, particularly when jaws act as a pair of scissors. Most food items being processed near the base of the bill, mechanisms have been developed which contribute to overcome the risk of breakage at the level of the prokinetic hinge because of a vertically oriented force there. PMID- 11725705 TI - Poaceae in the natural diet of the snail Helix aspersa Muller (Gastropoda, Pulmonata). AB - The natural diet of Helix aspersa was studied in two populations by analysing faeces. Picris echioides, Carduus tenuifloris, Urtica dioica, Galium molugo (Dicotyledons) and Poaceae (Monocotyledons) were the principal resources of the studied populations. The species of Poaceae ingested by Helix aspersa in July were determined by analysing the phytoliths present in the faeces. Festuca rubra, Dactylis glomerata and Bromus hordeaceus seemed to be attractive for the snails whereas Elytrigia repens was rejected. These results were convergent with the ingestion rates of Poaceae in the laboratory but assimilation efficiencies did not explain these choices. Poaceae with the highest energetic values were not preferred to others. The texture, the mineral and organic contents of the Poaceae may be responsible for the snails feeding choices. The importance of Poaceae for this species is discussed. PMID- 11725706 TI - Suspected side effects of a herbicide on dung beetle populations (Coleoptera: Scarabaeidae). AB - This paper addresses the association between use of a herbicide and anecdotal reports of reduced dung degradation and dung beetle populations. Dung beetles were monthly collected at two adjacent ranches in Mexico. Ranches were similar in area, elevation, exposition, soil, and vegetation, but differed in weed control. Ranch A controlled weeds manually, and ranch B controlled unwanted vegetation with applications of the herbicide Tordon 101M. The main species recovered on each ranch (Ataenius apicalis) was significantly more abundant at ranch A than at ranch B. Conversely, similar numbers of a second species, Ataenius sculptor, were recovered from both ranches. Three lines of evidence support the tentative conclusion that herbicide applications may be causing a decline in populations of A. apicalis on ranch B. First, the greatest reductions of A. apicalis were observed during periods of herbicide application. Second, A. sculptor, apparently little affected by these same herbicide applications, is active primarily during months without herbicide applications. Third, preliminary results of laboratory studies show that exposure to herbicide can impair reproductive function of the dung beetle Canthon cyanellus. PMID- 11725707 TI - [Optimization of culture conditions for in vitro rooting of argan (Argania spinosa L.)]. AB - The root system produced of in vitro organ plantlets is of poor quality and not efficient for the transfer to out-door conditions. To overcome such problems, experimentation was undertaken where the effects of growth regulators, nitrogen, sugar, activated charcoal and coconut fiber were tested on root induction and elongation. Modified Murashige and Skoog with half strength salt was used as a basal medium. Root induction (85%) with a mean of 16 roots per explant was obtained when shoots were grown, under dark conditions for 14 days, with a combination of two auxins (IBA and NNA), added at equal concentrations (5 mg.L 1). Secondary roots, 10 cm long, were initiated in 12% of the cultures in presence of 5 g.L-1 activated charcoal. Further improvements in the growth of the primary and secondary roots were obtained when semi-solid medium was substituted with a substrate composed of coconut fibers (80 g) mixed with semi-solid medium (35 mL) and agar (2.5 g.L-1). PMID- 11725708 TI - [Prediction of improvement of left ventricular systolic function by dobutamine echocardiography after recent myocardial infarction]. AB - Left ventricular ejection fraction is a major prognostic factor of ischaemic heart disease. In the early phase of myocardial infarction, part of the myocardium may be stunned and responsible for marked segmental wall dysfunction which is potentially reversible. The authors studied the potential of low dose dobutamine echocardiography to predict secondary improvement of left ventricular systolic function in 21 patients with recent inaugural myocardial infarction without primary angioplasty. All patients were treated and the investigation was carried out up to 20 micrograms/Kg/min of dobutamine without unwanted side effects or myocardial ischaemia. The detection of viability by this method was associated with improved wall motion of the affected segments in 74% of cases, most of which had benefited from myocardial revascularisation at control echocardiography performed 8 weeks later. If 4 or more segments were estimated to be viable initially, the left ventricular ejection fraction improved to a value comparable to that obtained at a dosage of 20 micrograms/Kg/min of dobutamine. On the other hand, there was no secondary improvement in 76% of segments estimated to be non-viable whether or not they had been revascularised. The sensitivity, specificity, positive and negative predictive values of low dose dobutamine echocardiography for prediction of myocardial recovery after recent infarction were respectively 71, 79, 74 and 76%. The results of this investigation show prognostic value and could be an aid to the decision concerning revascularisation of patients not having undergone primary angioplasty. PMID- 11725709 TI - [Management of dyslipidemias diagnosed in general practice in France--The PRAGMA Study]. AB - Several studies have reported the penetration and impact of national and international recommendations on the management of dyslipidaemia, a major cardiovascular risk factor. Most of them were carried out on patients participating in clinical trials or on in-hospital cases. The PRAGMA study was developed in order to evaluate management of this condition in general practice, at the heart of the health care system. From September to December 1998, 1,717 general practitioners were chosen randomly and included 6,623 patients considered to have a lipid disorder. In this sample, the prevalence of the main risk factors was as follows: hypertension: 39.6%, diabetes: 11.6%, obesity: 19.6%, past or present smokers: 33.8%. The main lines of management consisted in prescribing lipid lowering drugs (96.6%) with dietary recommendations (95.8%) and a fall lipid profile (59.9%). The main factors spontaneously cited by the general practitioners as being decisional were: the total cholesterol level (47.8%), diet (40.8%), body weight (29.4%) and drug therapy (19.2%). The cardiovascular risk factors were rarely taken into account in their totality. These results suggest that the management of dyslipidaemia patients by general practitioners is far from being optimal. Efforts should be made to change attitudes to take into consideration the global cardiovascular risk factors of patients with lipid disorders. PMID- 11725710 TI - [Representative observations of therapies in arterial hypertension]. AB - An enquiry was conducted by 125 general practitioners in 1998 concerning 1,204 hypertensive patients (55.5% women, 71% aged 60 or over, 54% with normalised blood pressures). At the first visit (inclusion) the most frequently described therapeutic classes were: diuretics (19%), betablockers (19%), ACE inhibitors (14%), angiotensin II inhibitors (7%). The use of fixed associations was uncommon except for that of ACE inhibitors with diuretics (10.5%). The incidence of normalisation of blood pressure was the same in all therapeutic classes, about 55%, comparable to reported data; however, 42% of non-normalised patients were on monotherapy. Only 7.4% of patients had their treatment changed at inclusion: 3.8% of normalised and 11.2% of non-normalised hypertensives. This low incidence of change of treatment is partially explained by the patients' own opinion about the treatment of their hypertension. In fact, the majority of therapeutic changes is observed when the patient and the doctor both concur. This data on the treatment of hypertension confirms the respective usage of the main therapeutic classes but the limited number of treatment changes at inclusion does not allow yet assessment of these new choices. One year follow-up of this cohort to the second phase of results should provide important information with respect to this point. PMID- 11725711 TI - [Investigation of fetal arrhythmias by simultaneous recording of ascending aortic and superior vena caval blood flow]. AB - In a period of 18 consecutive months, all the foetus referred to our Foetal Cardiology Unit for investigation of arrhythmia were systematically assessed by M mode echocardiography and simultaneous recording of blood flow in the superior vena cava and the aorta (SVC/Ao). This study was undertaken to compare the performance of these two approaches. The foetus were classified into three groups according to the arrhythmia diagnosed: Group 1: irregular arrhythmias, Group 2: bradycardias, Group 3: tachycardias. A surface ECG was recorded in all the neonates in whom the arrhythmias persisted. In Group 1, including 50 cases of extrasystoles (49 atrial and 1 ventricular), M mode echo and the Doppler provided the diagnosis in 42 and 47 cases respectively. This difference was not statistically significant. Group 2 comprised four cases of bradycardia (2 blocked atrial bigeminy, 2 complete atrioventricular blocks); the two methods provided the diagnosis in all 4 cases. Group 3 comprised 11 cases including 7 supraventricular tachycardias (SVT), 2 flutter, 1 chaotic atrial rhythm and 1 ectopic junctional rhythm. Complete analysis of these arrhythmias was possible by M mode in 4 cases and by Doppler in all cases. This difference was significant. The distribution of the 7 cases of SVT with respect to the duration of the ventriculoatrial interval was possible by M mode in 2 cases and in all cases by Doppler. This was a decisive factor in the choice of antiarrhythmic therapy. The authors conclude that Doppler and M mode are two echocardiographic approaches which are equally effective in the investigation of foetal atrial extrasystoles and probably of sustained foetal bradycardia. However, Doppler recording of SVC/Ao gives a more detailed and precise diagnosis of more complex foetal arrhythmias. PMID- 11725712 TI - [Anticoagulant therapy during pregnancy in patients with mechanical valvular prostheses]. AB - Anticoagulant therapy (heparin or oral anticoagulants) during pregnancy in patients with mechanical valvular prostheses should take into consideration the respective risks of the usage of these two products: thromboembolic for the mother, congenital malformations for the children and haemorrhagic for them both. Basically, heparin therapy is useful by limiting the risk of foetal complication, but it increases maternal risk, especially related to thromboembolism. The risks of oral anticoagulants are concentrated in the second half of the first trimester (coumarin-related embryopathies) and in the peri-partum period (maternal and foetal haemorrhage). There is general agreement to use oral anticoagulants outside these periods, during the second trimester and most of the third. During the first trimester, the European and North American Recommendations make provision for parental consultation. Some authors recommend oral anticoagulants even during the first trimester because of the scarcity of coumarin-related embryopathies. In fact, the incidence greatly increases when the dosage required is over 5 mg of coumarin per day. Others continue anticoagulation until the end of pregnancy, stopping 48 hours before delivery, either vaginal or by caesarean section. These attempts to avoid heparin therapy are related to difficulties of obtaining stable anticoagulation with non-fractionated heparin which increases the risk of thromboembolism, especially prosthetic valve thrombosis. Low molecular weight heparin, which is widely used in venous and arterial thromboembolic disease, including during pregnancy, is not yet recommended for usage in pregnant women with mechanical valvular prostheses because there are not results from large series concerning their efficacy and safety in this particular indication. In the absence of randomised studies in pregnancy, the renewal of small series in specialised centres will probably open up the field for their usage in the years to come. PMID- 11725713 TI - [Hypertension in pregnancy]. AB - Hypertension occurs in 10 to 15 p cent of pregnancies. Among them, 10 to 20% also have proteinuria. This situation defines preeclampsia, and involves a serious threat on foetal and even maternal prognosis. Presence of the hepatic (HELLP) syndrome still severely worsens the prognosis. Pathophysiology of preeclampsia is based on a very early abnormality of placentation, leading to insufficient blood supply to the foeto-placental unit. At the maternal level, the main consequence of placental ischemia is diffuse endothelial dysfunction, responsible for systemic vasoconstriction and clotting abnormalities. In such a context, merely lowering blood pressure with drugs is quite inefficient, or even harmful. The prognosis of this disease is mainly related to the pertinence of obstetrical management. An early preventive strategy is the most logical approach of preeclampsia, its modalities remain under discussion. Hypertension has a high recurrence rate on subsequent pregnancies. It is most often linked to a high global vascular risk level, therefore many of those patients will become permanent hypertensives in the near future. PMID- 11725714 TI - [Ablation of atrial fibrillation: where are we now?]. AB - Atrial fibrillation is the commonest arrhythmia. Besides the risk of complications, a significant number of patients remain symptomatic despite the different anti-arrhythmic drugs currently available. The only curative treatment is by surgery or catheter ablation. Since 1994, several approaches have been developed based on two main concepts: modification of the arrhythmogenic substrate by linear lesion to prevent the perpetuation of the arrhythmia and ablation of the foci initiating the atrial fibrillation. The later approach is the most popular one at the moment because the concentration of foci at the site of the pulmonary veins makes it possible to isolate them relatively easily. The presence of atrial foci in some patients complicates matters and limits the success rate to 70%. Despite these limitations and with an acceptable rate of complications, this approach appears preferable to His bundle ablation in young patients with symptomatic paroxysmal atrial fibrillation resistant to antiarrhythmic therapy. PMID- 11725715 TI - [Self monitoring versus ambulatory blood pressure monitoring]. AB - Blood pressure is a variable parameter for which the isolated determination cannot be considered as a characteristic of the subject. Therefore, the clinical measurement of blood pressure constitutes the method of reference but presents limits and causes of errors which pose a problem for its validity in evaluating the average blood pressure level in certain patients. In order to overcome the limits of occasional measurement, different methods of blood pressure measurement have been proposed. Among these methods, self monitoring of blood pressure by the patient and 24 hour ambulatory blood pressure monitoring (ABPM) are the most used. Each of these methods presents advantages and disadvantages, indications and limits of use. The information obtained by each of them is of a different nature: these methods are not substitutes but are complementary. Self monitoring and ABPM seem to evaluate the pressure load better than clinical monitoring and are better correlated to the organic effects of hypertension and thus the cardiovascular morbidity and mortality, ABPM in particular. However, it remains to be clarified whether the use of these methods in the evaluation of cardiovascular risk and the therapeutic management of the hypertensive can improve the long term cardiovascular prognosis. PMID- 11725716 TI - [Infectious endocarditis in France today]. AB - Infectious endocarditis (IE) is an uncommon condition carrying a relatively high mortality and morbidity. Two epidemiological studies, undertaken eight years apart, provide data allowing an appreciation of changes in the epidemiological and clinical profiles of IE. They show a progressive increase in the age of patients affected by IE and a decrease in percentage of IE on native pathological valves and valvular prostheses, compensated by the increase in incidence of IE occurring in patients with no known underlying cardiac disease. Moreover, there has been a change in microbiological profile, with the emergence of Streptococcus bovis and increase in staphylococcal IE, a decrease in IE due to oral streptococci and an improvement in microbiological diagnosis with negative blood cultures. Finally, from the therapeutic point of view, surgery is used more frequently and at an earlier stage of the disease. The global mortality of IE at the end of the initial hospital period remains 16%. PMID- 11725717 TI - [Paroxysmal atrial fibrillation in community medicine: management intentions and cost estimates]. AB - This study was undertaken to describe the declared management of atrial fibrillation in community medicine in France for patients between 50 and 80 years of age, and to estimate the total cost of treatment for Society in the year 2000. A questionnaire was sent to a sample of 306 private sector cardiologists, representative for their geographic location: 100 replies were received. Only 10% of cardiologists prescribed a treatment in patients without cardiovascular precedents who had a single regressive episode of atrial fibrillation and three quarters of them advised follow-up. This management cost an average between 228 and 296 Euros/year. When the episode of fibrillation was persistent at the time of consultation, 82% of practitioners prescribed antiarrhythmic reduction (amiodarone) after antithrombotic treatment (oral anticoagulants) followed by an antiarrhythmic therapy (class lc mainly) with an average cost of 659 to 783 Euros/year. In patients with previous cardiovascular disease, when a first episode of atrial fibrillation was present at the time of consultation, 92% of cardiologists prescribed pharmacological reduction with amiodarone, followed by long-term therapy when successful for an average cost of 755 to 1,092 Euros/year. All cardiologists requested systematic blood tests to search for thyroid complications of amiodarone with an average cost of 59 Euros/year. The costs were high, especially in chronic and recurrent pathology: the cost of treatment of the first two episodes of atrial fibrillation in the over 65 age group was 305 million Euros/year. PMID- 11725718 TI - [Malignant non-Hodgkin's lymphoma presenting with cardiac tamponade]. AB - Malignant non-Hodgkin's lymphoma rarely involves the heart. However, all cardiac structures may be invaded, although the myocardium and pericardium are affected more often than the valvular structures. Diagnosis is important because treatment must be adapted. The cardiac signs are usually non-specific and lymphomas presenting in this way have only occasionally been reported. The authors report two cases in which malignant non-Hodgkin's lymphoma presented with cardiac tamponade. PMID- 11725719 TI - [Valvular locations in Whipple's disease: report of two cases and review of the literature]. AB - Whipple's disease is a consequence of an infection due to a bacteria called Tropheryma whippelii. Endocarditis is frequently described in post mortem studies on this disease but the diagnosis in living patients is uncommon. PMID- 11725720 TI - [Cardiogenic shock in an infant due to rupture of an aneurysm of the sinus of Valsalva into the left ventricle]. AB - The authors report the case of a 15 month-old infant admitted to the intensive care unit for cardiogenic shock due to rupture of an aneurysm of the right anterior sinus of Valsalva into the left ventricle, associated with massive aortic regurgitation. The patient underwent a Ross procedure with resection of the aneurysm. This is a very rare condition because rupture of the aneurysm into the left ventricle has not been previously reported in a young girl to the best of the authors' knowledge. Moreover, a Ross procedure has not been previously described in this indication. The case illustrates the diagnostic difficulty associated with this pathology and underlines the necessity of thinking of this diagnosis in infants with appearances of dilated cardiomyopathy and aortic regurgitation. PMID- 11725721 TI - Molecular mechanisms of adipocyte differentiation. PMID- 11725722 TI - Regulation of leptin production. AB - Fat mass is the primary determinant of serum leptin in humans with energy intake and gender also having significant effects. Gender influences leptin production through the reproductive hormones. Glucose metabolism links food intake to leptin production and hexosamine biosynthesis appears to play a significant role in this process. Catecholamines inhibit leptin production and the sympathetic nervous system has been proposed to be the efferent arm of the leptin signal transduction pathway between adipose tissue and the central nervous system. Additional regulators of leptin production include glucocorticoids, cytokines and agonists of PPAR gamma. In addition to adipose tissue, leptin is produced in several other places including placenta, bone marrow, stomach, muscle and perhaps brain, thus increasing the number of potential regulatory roles for this hormone. Future work will be needed to fully elucidate the mechanisms regulating leptin synthesis/release in each tissue as well as its regulatory functions. PMID- 11725723 TI - Obesity in the Pimas. PMID- 11725724 TI - Pediatric obesity. AB - Pediatric obesity can be defined as BMI > 95th percentile for age and sex from large surveys that were carried out in the past. Using these cut points, over 10% of all children and adolescents are obese, and another 10% are overweight (BMI > 85th percentile). Obesity in childhood is associated with many immediate consequences, including orthopedic, neurologic, pulmonary, gastroenterologic, endocrinologic, metabolic, and cardiovascular disorders. Psychosocial and economic problems also are seen in obese adolescents. Long-term consequences of pediatric obesity include risks for cardiovascular disease and death that are independent of adult body weight. The differential diagnosis of pediatric obesity includes many multi-system genetic syndromes and a limited number of endocrine disorders that include abnormalities in the leptin signaling pathway, but most children with excess body weight do not have an established diagnosis. Treatment is best carried out at specialized centers, and usually involves a program of behavioral modification, caloric restriction, increased activity, and decreased sedentary behaviors. Medication for pediatric obesity cannot be recommended at present outside the context of clinical trials. Gastric bypass surgery is effective, but should be considered a last resort for the child with life threatening complications of obesity. PMID- 11725725 TI - The beta 3-adrenergic system and beta 3-adrenergic agonists. PMID- 11725726 TI - Does brown adipose tissue (BAT) have a role in the physiology or treatment of human obesity? PMID- 11725727 TI - Drug treatment of obesity. AB - At present only two drugs are approved for long-term treatment of obesity. Sibutramine inhibits the reuptake of serotonin and norepinephrine. In clinical trials it produces a dose-dependent 5-10% decrease in body weight. Its side effects include dry mouth, insomnia, asthenia, and constipation. In addition, sibutramine produces a small increase in blood pressure and pulse that is a contraindication to the use of this drug in some individuals with heart disease. Xenical is the other drug approved for long-term use in the treatment of obesity. It works by blocking lipase and thus increasing the fecal loss of triglyceride. One valuable consequence of this mechanism of action is the reduction of serum cholesterol that averages about 5% more than can be accounted for by weight loss alone. In clinical trials it produces a 5-10% loss of weight. Its side effects are entirely due to undigested fat in the intestine that can lead to increased frequency and change in the character of stools. It can also lower fat-soluble vitamins. The ingestion of a vitamin supplement before bedtime is a reasonable treatment strategy. The effect on weight loss during long-term trials with these two drugs is shown in Figs 7 and 8 above. Also in this figure is data on phentermine used in trials of six months or more. Although there were differences in mean weight losses with these drugs, when the placebo effect was taken into account they all had a surprisingly similar magnitude of weight loss. PMID- 11725728 TI - Sentinel lymph node detection and microstaging in vulvar carcinoma. AB - OBJECTIVE: To determine the efficacy of using complementary techniques for detecting sentinel lymph nodes (SLNs) in vulvar carcinoma and to evaluate the utility of microstaging techniques. STUDY DESIGN: Patients with invasive vulvar carcinoma underwent sentinel lymph node detection (SLND) using preoperative lymphoscintigraphy, intraoperative isosulfan blue dye injection and an intraoperative hand-held gamma-detecting probe. Eleven patients were included and a total of 16 groins evaluated. Sentinel nodes identified were excised, bisected and examined in surgical pathology using hematoxylin and eosin (H&E) staining. Pathologically negative SLNs were subjected to additional microstaging via serial sectioning and immunohistochemical staining for cytokeratin. Surgical management of the vulvar cancer and extent of inguinal-femoral lymphadenectomy were individualized based on clinicopathologic parameters, including depth of invasion, location of the tumor and patient performance status. RESULTS: Lymphoscintigraphy, dye and gamma-detector methods led to the total detection of 16, 19 and 17 SLNs, respectively. In two cases the isosulfan blue dye assisted in the isolation of an additional sentinel node over that of the gamma probe. Each method individually identified SLNs in 10/11 patients (91%). A total of 19 sentinel nodes were isolated. One SLN (5%) was positive for metastatic disease using H&E staining. Of the 18 negative SLNs, 2 (11%) had micrometastases (< 0.2 mm) upon serial sectioning and immunohistochemical staining. CONCLUSION: Combined modality mapping enhances detection of SLNs in vulvar carcinoma. Histologic microstaging improves the detection of micrometastases within SLNs. PMID- 11725729 TI - Women's decision-making determinants in choosing uterine artery embolization for symptomatic fibroids. AB - OBJECTIVE: To determine what symptoms of leiomyomata uteri prompted women to seek uterine artery embolization (UAE) and what factors were most frequently cited in the decision making leading to choosing UAE over other treatments. STUDY DESIGN: Eighty-four consecutive women with symptomatic leiomyoma presenting for UAE completed a questionnaire that inquired about their pelvic symptoms and the issues that were important in their decision to request UAE. All subjects previously had been told that they were surgical candidates. RESULTS: Pelvic symptoms that the 84 women most frequently noted were bleeding (n = 61), anemia (41), pelvic pain (29), frequent urination (24) and pelvic pressure (21). The majority of women (78) reported significant worry about their health from the fibroids, and (72) reported that the symptoms caused daily discomfort. Although the majority of women wanted a treatment that would give permanent relief of symptoms and thought UAE would do this, other factors frequently cited in the decision making included quality-of-life reasons, such as the desire to avoid adverse effects of other treatments (76), anticipated prolonged postoperative recovery from surgery (70) and avoiding surgery (66). Many women considered the uterus an important female organ, believed that the uterus was a source of femininity (33), stated that the uterus was necessary to maintain self-image (49) and reported that the uterus was necessary to maintain sexual image (49). CONCLUSION: In this cohort of women with symptomatic leiomyomas, treatment preferences did not interfere with the current lifestyle. In addition, the uterus was considered a source of femininity and sexuality. It is not clear whether women requesting UAE differ from women requesting surgical intervention in terms of how they assess the importance of the uterus, but these data suggest that many women still consider the uterus an important aspect of their femininity and that those seeking nonsurgical options should be thoroughly counseled about uterine function and how it relates to sexuality. PMID- 11725730 TI - Bone mineral density in young, hypothalamic oligoamenorrheic women treated with oral contraceptives. AB - OBJECTIVE: To evaluate whether decreasing doses of ethinyl estradiol affect bone loss related to hypothalamic amenorrhea. STUDY DESIGN: Sixty-four women with hypothalamic oligoamenorrhea were allocated to two therapy groups: group A (n = 24) received an OC containing 0.030 mg of ethinyl estradiol (EE) and 0.15 mg of desogestrel. Group B (n = 22) received an OC containing 0.020 mg of EE and 0.15 mg of desogestrel. Eighteen women who did not wish to use hormonal therapy constituted the control group (C). Calcium, phosphate and osteocalcin were measured basally and at 6 and 12 months of follow-up. Bone mineral density at the lumbar spine was determined before initiation of the study and at 12 months by dual energy X-ray absorptiometry. RESULTS: Serum calcium, phosphate and osteocalcin were significantly reduced by both active treatment regimens, whereas no differences were observed in the control group. Bone mineral density at 12 months showed an increase in both therapy groups (A, 2.4%; B, 2.5%), while group C showed a significant decrease (1.2%, P < .05). CONCLUSION: Both doses of EE were equally effective in preventing bone loss related to hypoestrogenism in hypothalamic oligoamenorrheic subjects. PMID- 11725731 TI - Effectiveness of thin-layer preparations vs. conventional Pap smears in a blinded, split-sample study. Extended cytologic evaluation. AB - OBJECTIVE: To further evaluate the effectiveness of the AutoCyte PREP thin-layer slide preparation (TriPath Imaging, Inc., Burlington, North Carolina) as compared to conventional Pap smears. STUDY DESIGN: A split-sample, blinded evaluation of matched thin-layer preparations and conventional smears from 2,438 patients was performed. This material was enriched by including 260 cases of high grade squamous intraepithelial lesions (HSILs) and cancer cases from an earlier study. Many of these cases were difficult to diagnose, containing very few abnormal cells on one or both matching slides. The preparations were evaluated multiple times by both thin-layer-inexperienced and -experienced cytology professionals to better compare performance related to preparation quality alone. RESULTS: The initial evaluations of the slides by personnel with only brief training in thin layer interpretation demonstrated equivalent performance for the two preparations. The reevaluation study by cytology professionals with several months of thin-layer experience demonstrated a statistically significant improvement in detection of both LSIL and HSIL lesions using AutoCyte PREP slides. There was also a statistically significant improvement in the number of satisfactory samples using the AutoCyte PREP method. CONCLUSION: The study demonstrated that the AutoCyte PREP thin-layer slide preparation is at least equivalent to conventional Pap smears in the detection of LSIL and HSIL, even when evaluated by cytology professionals who have been newly trained in the thin layer method and that, with increased experience, the thin-layer AutoCyte PREP slide preparation method showed a statistically significant improvement in disease detection. PMID- 11725732 TI - Increased nitric oxide in peritoneal fluid from women with idiopathic infertility and endometriosis. AB - OBJECTIVE: To verify whether nitric oxide in peritoneal fluid is associated with endometriosis and infertility. STUDY DESIGN: Twenty-five women with idiopathic infertility and 38 with endometriosis were recruited, and 18 cases of uterine myomata and 2 cases of ovarian cyst served as controls. Peritoneal fluid samples were aspirated from the pouch of Douglas during laparoscopy or laparotomy. Metabolites of nitric oxide (nitrite and nitrate) in peritoneal fluid were determined by a method using nitrate reductase and the Griess reaction. RESULTS: Peritoneal concentrations of nitrate/nitrite in both infertile women (42.02 +/- 12.98 mmol/L) and patients with endometriosis (41.75 +/- 16.42 mmol/L) were significantly higher than that in controls (33.96 +/- 13.07, P < .05 for both). No significant difference in peritoneal nitrate/nitrite level was found between infertile women and patients with endometriosis (P > .5). Peritoneal levels of nitrate/nitrite were comparable among patients with endometriosis at different stages (P > .5). Patients with endometriosis had more peritoneal fluid than controls and idiopathic infertile women, while controls and idiopathic infertile women had comparable amounts of peritoneal fluid. CONCLUSION: An increased peritoneal level of nitric oxide is a common alteration in endometriosis, endometriosis-associated infertility and idiopathic infertility and may be associated with the pathogenesis of these diseases. PMID- 11725733 TI - Three protocols for monitoring follicle development in 587 unstimulated cycles of in vitro fertilization and intracytoplasmic sperm injection. A comparison. AB - OBJECTIVE: To test the adequacy of unstimulated cycles for intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) and to evaluate implantation and pregnancy rates in three monitoring protocols. STUDY DESIGN: A retrospective chart review of 587 patients undergoing IVF and ICSI in unstimulated cycles was performed. In the first group (protocol A), all cycles were monitored by ultrasound only. Human chorionic gonadotropin (hCG) was given when the mean follicle diameter reached 18 mm. In protocol B, hCG was given when serum estradiol (E2) or follicle diameter reached the critical value (0.91 nmol/L and 18 mm). With a smaller follicle diameter, the E2 level had to be higher, and vice versa. In protocol C, hCG was administered when the serum E2 was > 0.49 nmol/L and follicle diameter at least 15 mm. Cycles with positive luteinizing hormone in urine before hCG was given were cancelled. RESULTS: The cancellation rate was lower in protocol C (33/335, 9.8%) than protocol B (42/151, 27.8%) and A (41/101, 40.5%). In protocol C the pregnancy rate per cycle was higher with IVF (n = 219) and ICSI (n = 116) cycles (10.5% and 12.1%) than when protocol B was used (3.8% and 4.3%). The pregnancy rate per transfer was highest when protocol C was used in the IVF (23/105, 21.9%) and ICSI group (14/53, 26.4%). CONCLUSION: Unstimulated cycles monitored by serum E2, urinary luteinizing hormone and ultrasound can produce an acceptable pregnancy rate after IVF and ICSI. PMID- 11725734 TI - Preinduction cervical ripening. A randomized trial of intravaginal misoprostol alone vs. a combination of transcervical Foley balloon and intravaginal misoprostol. AB - OBJECTIVE: To determine if the addition of a mechanical ripening agent (transcervical Foley balloon) to a pharmacologic agent (intravaginal misoprostol) improves the efficiency of preinduction cervical ripening. STUDY DESIGN: Singleton patients with an indication for delivery, unfavorable cervix (Bishop score < or = 5) and no contraindication to labor were randomly assigned to two groups: misoprostol alone (25 micrograms intravaginally every 3 hours for no more than 12 hr) or combination therapy (25-French transcervical Foley balloon inflated to 50 mL of sterile water with identical intravaginal misoprostol dosing). All patients received a history and physical examination (including Bishop score), preripening ultrasound, electronic fetal heart rate and contraction monitoring (to rule out spontaneous labor and document fetal well being). Multiple variables of perinatal outcome were analyzed, including the main outcome variables of ripening-to-delivery time and cesarean section rate. RESULTS: During August 1998 to August 1999, 81 patients were randomized, 40 to misoprostol alone and 41 to combination therapy. There were no differences between the groups with respect to maternal demographics, preripening Bishop score, maternal complications, intrapartum intervention or neonatal outcome. The misoprostol group spent longer periods of time in active labor, and there was a trend for the combination group to require oxytocin for longer intervals. These findings did not significantly affect the total ripening-to-delivery time or cesarean section rate which were similar for both groups. CONCLUSION: The addition of mechanical ripening with a transcervical Foley balloon to intravaginal misoprostol did not improve the efficiency of preinduction cervical ripening. Mechanical and pharmacologic cervical ripening agents appear to act independently rather than synergistically. PMID- 11725735 TI - DeLee suction. Does it have clinical significance? AB - OBJECTIVE: To evaluate the null hypothesis that there would be no difference in neonatal outcome in infants who underwent DeLee suction vs. those who underwent bulb suction when thick meconium was present. This difference was defined as a one-day difference in length of stay, two-point difference in the five-minute Apgar score or significant difference in blood gases. STUDY DESIGN: A power calculation based on a one-day length of stay found that 115 patients would be needed in each arm. A cohort of 115 consecutive deliveries with the presence of thick meconium was reviewed during each time period when DeLee suction was utilized and then matched with deliveries that used bulb suction. RESULTS: Two hundred thirty charts were reviewed. Length of stay, one-minute Apgar score, five minute Apgar score, blood gases, gestational age and incidence of amnioinfusion were compared. There was no significant difference in any variable. Meconium aspiration syndrome in the period studied remained stable at < 1%. CONCLUSION: There was no significant clinical change in neonatal outcome with the abandonment of nasopharyngeal DeLee suction on the perineum when meconium was present. We recommend the use of bulb suction as routine obstetric practice even in the presence of thick meconium. PMID- 11725736 TI - Development of antibodies to topical bovine thrombin after abdominal hysterectomy. A case report. AB - BACKGROUND: Topical thrombin spray is frequently used as a hemostatic agent in surgery, particularly following cardiovascular, orthopedic and neurologic surgery. Acquired coagulation factor inhibitors to thrombin and factor V may develop following gynecologic surgery with repeat topical thrombin use. CASE: A 45-year-old woman underwent total abdominal hysterectomy/bilateral salpingo oophorectomy. Her past medical history included idiopathic thrombocytopenic purpura and Marfan's syndrome and past surgery included splenectomy and aortic valve replacement with a three-vessel coronary artery bypass graft. She was converted from Coumadin (DuPont Pharmaceuticals, Wilmington, Delaware) to heparin preoperatively, and heparin was stopped six hours before surgery, with normalization of her prothrombin time (PT) and partial thromboplastin time (PTT). Topical bovine thrombin spray was applied to the surgical field in anticipation of early resumption of heparin anticoagulation. Seven days after surgery, after recovering from some acute bleeding problems, the patient began to have elevation of her PT in spite of being off Coumadin for 10 days. Her PT and international normalized ratio (INR) continued to rise, reaching peak values of 31.4 and 6.99, respectively. A mixing study did not correct the problem, confirming the presence of coagulation inhibitors. The patient was given intravenous immunoglobulins and corticosteroids, and her PT/PTT and INR values slowly normalized. CONCLUSION: Acquired coagulation factor may develop following gynecologic surgery with topical thrombin use. This potential complication should be strongly considered in any patient prior to using topical bovine thrombin, especially those with a prior exposure or history of surgical procedures in which bovine thrombin is commonly used. PMID- 11725737 TI - Sepsis after Bartholin's duct abscess marsupialization in a gravida. AB - BACKGROUND: Little information exists regarding sepsis following marsupialization of a Bartholin's duct abscess. We report a gravida who became septic after marsupialization. CASE: A 30-year-old primigravida at 32 weeks' gestation underwent marsupialization of a Bartholin's gland abscess. Postoperatively, she developed fever with maternal and fetal tachycardia. She was admitted to the hospital and started on broad-spectrum antibiotics. Her temperature increased to 39 degrees C, and she became hypotensive. Blood work demonstrated evidence of disseminated intravascular coagulopathy. The patient was stabilized with aggressive fluid resuscitation, antibiotics, transfusion of blood products and oxygen therapy. Within 24 hours, the fever and coagulopathy resolved. She was discharged on postoperative day 5 and gave birth without complications at 38 weeks' gestation. CONCLUSION: Pregnant women undergoing marsupialization of a Bartholin's gland abscess should be considered at high risk and managed accordingly. PMID- 11725738 TI - Central hemodynamic monitoring in a woman with acute respiratory insufficiency after evacuation of a complete molar pregnancy. A case report. AB - BACKGROUND: The incidence of hydatiform moles in the United States is approximately 1 in 1,200 pregnancies. Acute respiratory insufficiency is a known complication of molar pregnancies, occurring in 8-11%. While there have been numerous case reports and retrospective studies describing respiratory complications following evacuation of hydatiform moles, only a limited number of reports provide data from central hemodynamic monitoring in patients with this complication. CASE: A 16-year-old, Hispanic woman, gravida 1, para 0, presented to the emergency room at 13 weeks' gestational age by menstrual dating with complaints of vaginal bleeding for two days. The serum quantitative beta-hCG level was 1 x 10(6) mIU/mL, and a bedside sonogram was consistent with hydatiform mole. After informed consent was obtained, the patient underwent dilation and suction curettage. Approximately five minutes after evacuation of the uterus was begun, the patient developed pulmonary edema in the setting of oliguria. A pulmonary artery catheter was inserted to determine the etiology of the edema. The initial pulmonary capillary wedge pressure was > 18 mm Hg, consistent with hydrostatic pulmonary edema. Volume overload in association with a reduced colloid osmotic pressure to wedge pressure gradient was primarily responsible for the pulmonary edema in this patient. CONCLUSION: The majority of case reports of pulmonary complications after evacuation of a hydatidiform mole were either presumed or documented to be due to trophoblastic pulmonary embolism. Thyrotoxicosis, fluid overload with dilutional anemia, preeclampsia, sepsis, hypoalbuminemia or a combination of these factors may be more common than trophoblastic embolization. PMID- 11725739 TI - Successful enucleation of a necrotizing fibroid causing oligohydramnios and fetal postural deformity in the 25th week of gestation. A case report. AB - BACKGROUND: In approximately 2% of pregnancies, uterine fibroids are detected. In 10% of diagnosed cases some complications are to be expected during pregnancy and/or delivery. CASE: A 32-year-old woman presented in the 25th week of gestation with a fibroid causing oligohydramnios and fetal postural deformity. A laparotomy and conservative myomectomy were performed with the hope of preserving the pregnancy. Ultrasound examinations performed postoperatively showed that the fetal postural deformity had ended and that the pace of fetal development had returned to normal. A healthy, male newborn weighing 3,600 g was born in the 40th week of gestation via cesarean section. CONCLUSION: The medical literature generally agrees upon conservative therapy; however, there are indications for laparotomy performed during the course of pregnancy complicated by a fibroid. In this case the fetal postural deformity, retardation in development and oligohydramnios, recognized preoperatively, provided indications for laparotomy. The success of the operation was established by postoperative sonography, which showed cessation of the fetal postural deformity and registered a normal volume of amniotic fluid. PMID- 11725740 TI - Serum tryptase analysis in a woman with amniotic fluid embolism. A case report. AB - BACKGROUND: Recent studies have noted a striking similarity between amniotic fluid embolism (AFE) and anaphylaxis. Serum tryptase levels may therefore serve as a marker of mast cell degranulation in AFE cases. CASE: A 40-year-old woman, gravida 6, para 4, experienced the acute onset of facial erythema, eclampsia-type seizures, severe hypoxia, cardiac arrest and disseminated intravascular coagulation while in early active labor. The patient was declared dead 37 minutes after the onset of resuscitative efforts. At autopsy, fetal squames were found within the pulmonary tree, uterine blood vessels and brain. A peripheral venous blood specimen, obtained approximately one and a half hours postmortem, revealed a tryptase level of 4.7 ng/mL (normal, < 1). CONCLUSION: An elevated serum tryptase level, in conjunction with our patient's clinical history, adds further supporting evidence to the concept of AFE as an anaphylactoid syndrome of pregnancy. PMID- 11725741 TI - Diagnosis of culture-negative female genital tract tuberculosis with peritoneal involvement by polymerase chain reaction. AB - BACKGROUND: Female genital tract tuberculosis (TB) is a common cause of infertility in developing countries. It is a paucibacillary form of the disease of which smears and cultures are usually negative. CASE: We were able to use polymerase chain reaction (PCR) amplification of Mycobacterium tuberculosis DNA to support a clinical and histologic diagnosis of a typical case of culture negative female genital tract TB. CONCLUSION: PCR may be a useful adjunct to diagnostic efforts in gynecologic tuberculosis. PMID- 11725742 TI - Balloon thermoablation in a woman with complex endometrial hyperplasia with atypia. A case report. AB - BACKGROUND: Endometrial ablation is an alternative to hysterectomy in patients with menorrhagia refractory to medical therapy. The histologic changes in the hyperplastic endometrium secondary to endometrial ablation are not well understood. CASE REPORT: A 44-year-old woman, gravida 4, para 4, had menorrhagia, was at high risk for medical treatment and underwent thermal balloon ablation. The dilatation and curettage specimen at the time of the procedure revealed complex hyperplasia of the endometrium with atypia. Subsequently a hysterectomy was performed, and the pathology specimen showed scarring, fibrosis and focal, weakly proliferative phase endometrium. There was no evidence of hyperplasia or malignancy. CONCLUSION: Endometrial ablation with a thermal balloon seems to have resulted in resolution of endometrial hyperplasia. However, hyperplasia of the endometrium should still be a contraindication to such a procedure because of the inability to accurately assess the endometrium after ablation. PMID- 11725743 TI - Intraplacental smooth muscle tumor. A case report. AB - BACKGROUND: Chorioangioma is the leading diagnosis for circumscribed placental lesion noted on ultrasound. Other rare intraplacental tumors may have a significant clinical impact on the mother and fetus. CASE: An intraplacental, hypoechoic area consistent with chorioangioma was noted on ultrasound examination at 17 weeks' gestation. The lesion remained stable in size throughout gestation, with no evidence of fetal hydrops or polyhydramnios. At term, a live-born male infant was delivered vaginally, followed by spontaneous expulsion of the placenta. Histologic examination of the placenta revealed an intraplacental, potentially malignant smooth muscle tumor. Six weeks postpartum, total abdominal hysterectomy and pelvic node sampling were performed. No residual disease was found on the final histologic examination. CONCLUSION: Although ultrasound differentiation is difficult, rare nontrophoblastic tumors should be considered in the differential diagnosis of placental masses. To our knowledge, this is the first report of such a lesion in the placenta. PMID- 11725744 TI - [Lower abdominal pain and urinary problems in a middle-aged woman]. PMID- 11725745 TI - [A long fallopian tube]. PMID- 11725746 TI - Controversial aspects of child abuse: a roundtable discussion. 43rd annual meeting, Society for Pediatric Radiology. PMID- 11725747 TI - Racism in medicine. Institutional racism in the BMJ. PMID- 11725748 TI - Racism in medicine. Regional arbitration panel may offer solution. PMID- 11725749 TI - Racism in medicine. We need action, not more evidence. PMID- 11725750 TI - Racism in medicine. Evidence base is needed. PMID- 11725751 TI - Racism in medicine. Environment for open, constructive debate is needed. PMID- 11725752 TI - Racism in medicine. Everyone should be treated with respect and dignity. PMID- 11725753 TI - Equity in the new NHS. Small groups should not undertake responsibilities on their own. PMID- 11725754 TI - Equity in the new NHS. Evidence cannot help in all situations. PMID- 11725755 TI - Drugs for Alzheimer's disease. More effective agents are needed. PMID- 11725756 TI - Drugs for Alzheimer's disease. Guidelines for prescribing cholinesterase inhibitors in Australia are similar to those in UK. PMID- 11725757 TI - Antihypertensive treatment and compliance. Pharmacological properties of antihypertensive drugs research need to be acknowledged. PMID- 11725758 TI - Antihypertensive treatment and compliance. Non-adherence should be addressed first. PMID- 11725759 TI - Antihypertensive treatment and compliance. Study was too short and lacked active intervention. PMID- 11725760 TI - An unmerciful end. Decisions not to resuscitate must not be left to junior doctors. PMID- 11725761 TI - An unmerciful end. Society must show respect for people who are dying. PMID- 11725762 TI - President's Task Force to Improve Health Care delivery for Our Nation's Veterans. PMID- 11725763 TI - National Research Service Awards. Final rule. AB - The National Institutes of Health (NIH) is amending the regulations governing National Research Service Awards (NRSA) in order to incorporate changes necessitated by enactment of the Alcohol, Drug Abuse, and Mental Health Administration (ADAMHA) Reorganization Act of 1992, Public Law 102-321, and the National Institutes of Health Revitalization Act of 1993, Public Law 103-43. PMID- 11725764 TI - [Soluble transferrin receptor and erythropoietin in chronic disease anemia with or without iron deficiency]. AB - The regulation of transferrin receptor (RTF) is related to intracellular iron stores and with the soluble receptor is present in plasma. It has already been demonstrated that in iron deficiency anemia (IDA), receptor expression increases when iron stores decrease. In anemia of chronic diseases (ACD) it is difficult to establish the real iron status because of the influence exerted by inflammatory or infectious diseases on iron metabolism. We studied 30 healthy normal subjects and 42 anemic patients (hemoglobin less than 120 g/L) affected with ACD divided into two groups with and without iron deficiency, in order to establish the diagnostic value of measuring the soluble transferrin receptor (sRTF). We correlated erythropoietin (EPO) (as an erythropoietic stimulating factor) with the decreased hemoglobin values observed in both groups. The results were analysed with an ANOVA statistic test of one way analysis of variance, and there were no significant differences in sRTF values between the ACD groups with or without iron deficiency. The ratio log EPO vs hemoglobin showed a remarkably significant inverse correlation in both groups. We can conclude that sRTF levels are within the normal reference values in these patients and are not related to organic iron. Consequently, sRTF cannot be considered a good parameter for making a diagnosis of iron deficiency in chronic diseases. PMID- 11725765 TI - Reasonable charges for medical care or services. Interim final rule. PMID- 11725766 TI - Medicaid program; use of restraint and seclusion in psychiatric residential treatment facilities providing inpatient psychiatric services to individuals under age 21. Interim final rule; amendment and clarification with request for comment. AB - On January 22, 2001, we published an interim final rule with comment period (66 FR 7148) that established a definition of a "psychiatric residential treatment facility" that is not a hospital and that may furnish covered Medicaid inpatient psychiatric services for individuals under age 21. The interim final rule established standards for the use of restraints or seclusion that psychiatric residential treatment facilities must have in place to protect the health and safety of residents. In response to some of the concerns submitted in comments on that interim rule, this document clarifies what facilities are subject to the requirements of the interim final rule, modifies reporting requirements to facilitate HCFA monitoring, and amends staffing requirements applicable to restraints and seclusion. Due to the operational significance of these issues, amendment to the interim final rule is required by the May 22, 2001 effective date of the interim final rule. Without such amendments, we are concerned that substantial numbers of facilities would not be able to comply with certain requirements of our interim final rule, and that beneficiaries will suffer needless displacement from those facilities. We are also concerned that HCFA will not be able to timely obtain data necessary to monitor for situations involving jeopardy to program beneficiaries. We will accept comments on these amendments, and will address all comments on the interim final rule and these amendments at a later date. PMID- 11725767 TI - High volume sutureless intraocular lens surgery in a rural eye camp in india. AB - BACKGROUND AND OBJECTIVE: To describe the use of small incision sutureless cataract surgery (SISCS) that permits high-volume, high-quality, and low-cost surgery. PATIENTS AND METHODS: We compared the speed, safety, and visual results of 1190 surgical techniques performed in a public eye camp in India. Intraocular lenses were received by 97% of patients (1154). Surgical techniques used were SISCS, phacoemulsification, and standard extracapsular cataract extraction (ECCE) technique. RESULTS: Postoperatively, 60.0% attained uncorrected vision of 6/24 or better. There was little difference in visual results or complication rates among the three techniques. The most striking finding is the speed of SISCS, which enables experienced surgeons to perform the technique in 3.8 to 4.2 minutes. CONCLUSION: We believe this surgical technique is of major importance especially in developing nations facing enormous surgical volumes and limited resources. PMID- 11725768 TI - Subretinal, viscoelastic-assisted, endoscope-guided photothermal ablation of choroidal neovascular membranes by Erbium:YAG laser. AB - BACKGROUND AND OBJECTIVE: The efficacy of endoscope-guided Erbium:YAG laser, assisted by viscoelastic macular detachment, in subretinal ablation of choroidal neovascular membranes (CNVs) is evaluated. PATIENTS AND METHODS: A high repetition, midinfrared erbium:YAG laser was used subretinally to ablate CNVs in 2 patients. Ablation followed pars plana vitrectomy and macular retinal detachment using a viscoelastic substance, and was guided by a high resolution endoscope. RESULTS: Visual acuity improved from 6/90 to 6/20, six months after treatment in 1 patient, and from finger counting from 1 meter to 6/30, three months after treatment in the other. The membranes showed no angiographic signs of activity postoperatively. However, a zone of stippled hyperfluorescence appeared in both patients, corresponding with the macular detachment area. This gradually diminished in size in both patients. CONCLUSIONS: The surgical procedure was found effective in our preliminary research. The use of viscoelastic substance and endoscope is felt to contribute to the favorable outcome. The cause and significance of the retinal pigment epithelium damage are yet to be determined. PMID- 11725769 TI - Preserved human amniotic membrane transplantation in the treatment of primary pterygium. AB - BACKGROUND AND OBJECTIVE: To assess the outcome of simple excision with preserved human amniotic membrane transplantation in the treatment of primary pterygium. PATIENTS AND METHODS: A total of 59 eyes with primary pterygium underwent surgical excision. In Group 1, 28 eyes were treated with simple excision and preserved human amniotic membrane transplantation. In Group 2, 31 eyes were treated with bare sclera excision. These two groups were compared in recurrence, final appearance of the operation site, and complications. Patients were followed for at least 10 months. RESULTS: During a mean follow up of 14.9 months, we observed 3 (10.7%) recurrences in Group 1 and 20 (38.7%) recurrences in Group 2 (P:0.03). In Group 1, 20 (71.4%) eyes and 14 (45.2%) eyes in Group 2 had a satisfactory final operation site appearance (P:0.041). No serious complication was observed in both groups. CONCLUSION: Simple excision and preserved human amniotic membrane transplantation appears to be a safe and effective way of treating primary pterygium because of the lack of serious complications and a relatively low rate of recurrence. PMID- 11725770 TI - Traumatic wound dehiscence after corneal graft. AB - BACKGROUND AND OBJECTIVE: Any trauma to a corneal transplanted eye is likely to lacerate the corneal wound because of persistent wound weakness, even years after keratoplasty. We evaluate the risks and consequences of trauma after penetrating keratoplasty (PKP). METHODS: Records of 11 patients who had been treated in our department from 1992 to 1998 for traumatic wound dehiscence after PKP were reviewed for the type of insult, visual acuity, operative methods, and outcome. RESULTS: The period between corneal grafting and wound rupture ranged from two months to three years. In 9 patients, the sutures were still in place. All the ruptures occurred at the donor-recipient interface. In 5 patients, the rupture was in the inferior half of the wound, in another 5 patients in the superior half, and in 1 patient it was nasal. In all the patients, the extent of the rupture was over 120 degrees and the lens was effected. All the patients had posterior segment involvement, the extent of which was related to the visual outcome. One eye was enucleated, another eye regrafted, and 8 of the remaining 9 grafts were restored and remained transparent. CONCLUSIONS: Despite severe trauma, most of the grafts' clarity can be restored, and good visual results can be achieved. PMID- 11725771 TI - Optical coherence tomography in the diagnosis of subclinical serous detachment of the macula secondary to a choroidal nevus. AB - We present a case in which optical coherence tomography (OCT) aided in the diagnosis of subretinal fluid (SRF) secondary to a choroidal nevus. A patient with a raised lesion above the left optic disc and distorted central vision was referred to our unit because of suspected choroidal melanoma. Fundus fluorescein angiography and slit-lamp investigation did not indicate any areas of SRF; however, OCT showed serous detachment in the macular area. The lesion was treated with laser photocoagulation and, when reviewed at four weeks post-treatment, repeat OCT scans showed that the area of SRF had cleared. In this case OCT was able to detect a subclinical serous detachment in the macula. This may have important implications in the monitoring of patients with small suspicious lesions. PMID- 11725772 TI - Purtscher-like retinopathy after retrobulbar injection. AB - Purtscher-like retinopathy occurred in a 46-year-old Chinese male who received 5 mL of 2% lignocaine, retrobulbar injection, and underwent pterygium excision with conjunctiva graft. The visual acuity improved spontaneously from 20/120 on the first postoperative day to 20/20 within 8 weeks. The disc swelling, veinous tortuosity, retinal whitening, and hemorrhage resolved with time. However, relative afferent papillary defect and an arcuate visual field deficit persisted. The mechanisms of this Purtscher-like retinopathy are postulated. PMID- 11725773 TI - Extraocular muscle metastasis 16 years following primary prostate carcinoma. AB - An 83-year-old man with a history of localized prostate carcinoma treated 16 years prior with transurethral partial prostatectomy and radioactive seed implants presented with a 2-month history of right globe proptosis. Computed tomography demonstrated a right medial rectus mass. Serum prostate specific antigen (PSA) level was normal. Tumor biopsy revealed poorly differentiated carcinoma with diffuse staining of tumor cells with PSA and prostatic acid phosphatase. This case demonstrates that prostate carcinoma may metastasize to an extraocular muscle many years after treatment of the primary tumor and may not be accompanied by elevated serum PSA. Immunohistochemical stains are helpful in establishing the diagnosis. PMID- 11725774 TI - Aureobasidium pullulans scleritis following keratoplasty: a case report. AB - Fungal scleritis is a rare entity. A 50-year-old patient with culture proven Aureobasidium pullulans corneal ulcer underwent therapeutic keratoplasty. He developed scleritis 5 days following surgery. Although the patient had symptomatic improvement after antifungal therapy, surgical debridement, and cryotherapy, visual improvement was only marginal. Aureobasidium pullulans should be looked for as a cause for keratomycosis and scleromycosis, especially in tropical countries. PMID- 11725775 TI - Morphology of the trabecular meshwork three years after Erbium:YAG laser trabecular ablation. AB - Laser trabecular ablation (LTA) is an abinterno approach with the purpose to improve outflow facility by removing trabecular tissue and open Schlemm's canal. In a blind eye with secondary glaucoma following silicone oil surgery, Erbium:YAG LTA had been performed applying 12 neighboring single laser pulses (5-7 mJ pulse energy, 200 micros pulse duration) to the trabecular meshwork. Enucleation was performed three years after the procedure that did not achieve pressure control. Light-microscopy revealed neither marked scarring nor endothelial proliferation in the treated trabecular meshwork. However, most of the recognizable ablation craters failed to open Schlemm's canal. Although limited scar formation within the trabecular meshwork after LTA is a promising aspect, the present technique of Er:YAG LTA still needs technical and surgical improvements to guarantee reliable and reproducible openings of Schlemm's canal. PMID- 11725776 TI - Closure of chronic macular holes using passive aspiration to the edges of the macular hole. AB - Full-thickness macular holes are characterized by central dehiscence of neurosensory tissue and a reduction in visual acuity. According to Gass (1) and then Johnson and Gass, (2) full-thickness macular holes are caused by progressive vitreous cortex condensation, resulting in tangential traction with centrifugal displacement of photoreceptor elements at the fovea. PMID- 11725777 TI - Hyperbaric oxygen therapy for mitomycin C-induced scleral necrosis. AB - Intraoperative or postoperative use of mitomycin C is one of the treatment options to reduce the recurrence of pterygia. Scleral necrosis, a potentially blinding complication after mitomycin C use, has been among great concern in ophthalmic practice. A patient is presented in whom scleral necrosis developed 20 days after a pterygiectomy with bare sclera technique was performed, followed by the application of 0.4 mg/mL of topical mitomycin C eye drops for 3 weeks. After conjunctival flap failures the patient was treated with hyperbaric oxygen therapy. The sclera revascularized and returned to near normal thickness after 24 sessions of hyperbaric oxygen therapy. This case demonstrates that it is an effective, novel treatment. [Ophthalmic PMID- 11725778 TI - Retrieval of a dropped corneal button from a vitrectomized eye. AB - A successful retrieval of a dropped corneal button from an aphakic vitrectomized eye during penetrating keratoplasty is reported. A 52-year-old female patient underwent penetrating keratoplasty for aphakic corneal edema in a vitrectomized eye. The donor button inadvertently dropped into the vitreous cavity and was retrieved successfully. The dropped corneal button was visualized after focusing the operating microscope on the retina. The visualization significantly improved after aspiration of the fluid from the vitrectomized eye. The donor button was picked up by McPherson's forceps and subsequently sutured to the recipient's cornea. Corneal surgeons need to be aware of this rare complication in aphakic vitrectomized eyes and its retrieval after accurate visualization. PMID- 11725779 TI - Endoscopic goniotomy with anterior chamber maintainer: surgical technique and one year results. AB - The surgical technique of endoscopic goniotomy (EG) using the anterior chamber maintainer (ACM) for congenital glaucoma is evaluated, and one-year follow-up data are presented. Endoscopic goniotomy was performed in 12 eyes of 7 patients. A double-port-special goniotomy knife mounted on the endoscope's probe inserted through the first incision and ACM through the second incision-EG technique was used in 6 eyes of 3 patients, while a three-port-knife, endoscope probe, and ACM inserted through separate incisions-technique was preferred in the remaining 6 eyes of 4 patients. EG of approximately 240 degrees could be done without major complications in all eyes. At the end of the average follow-up period of 14.2 +/- 9.7 months, the mean intraocular pressure was reduced from 38.3 +/- 6.9 mm Hg to 17.6 +/-, 2.8 mm Hg (P = 0.002), the average number of glaucoma medications from 2.1 +/- 0.3 to 0.3 +/- 0.5 (P = 0.001), and the mean cup/disk ratios from 0.84 +/ 0.11 to 0.79 +/- 0.14 (P = 0.014), while there was no statistically significant change in the average corneal diameter (P = 0.16). Therefore, endoscopic goniotomy with ACM was found to be an effective treatment modality for congenital glaucoma. PMID- 11725780 TI - Sinus fracture--phacoemulsification technique for dense cataracts. AB - A method of performing phacofracture of hard nuclei is presented. Following a central horizontal fracture into 2 hemisections, the phacoprobe is plunged into the inferior hemisection to create a sinus within its dense wall. The chopper/second instrument is placed into the sinus and force is applied along the lateral walls of the sinus. The phacoemulsification probe breaks it and splits the inferior hemisection into two quadrants. A similar method is employed for the superior-nuclear hemisection. PMID- 11725781 TI - Development and characterization of edible peanut protein films. AB - In this work, novel edible films were developed from peanut protein with sorbitol as a plasticizer. The peanut protein was extracted from peanut flour. Two groups of films were prepared. One group contained 10% in weight of natural oil/fat of the peanuts, while the other group contained no oil/fat. Six types of films with 37.5%, 44.5%, and 50% sorbitol content by weight, with and without fat, were chosen for mechanical properties characterization and microscopic observations. It was found that with the increase of sorbitol content, higher tensile strength and higher Young's modulus were achieved for the films without fat. However, films with fat exhibited maximum strength and modulus at 44.5% sorbitol content. Relationships between the mechanical properties and the microscopic features of these films were established. It was found that the higher the content of the sorbitol, the more evident the crystalline structure. Grant numbers: NCC9-51. PMID- 11725782 TI - Electrochemical removal of ammonium ions from a bioreactor effluent. AB - Ammonium ions are byproducts of the oxidation of nitrogen-containing substances occurring in the initial treatment steps of water recovery systems. Removal of ammonium ions from the effluent stream from 1000 ppm to less than 0.25 ppm is an imperative need as a part of the space life support infrastructure. Drawbacks associated with processes proposed in the past include the generation of a secondary waste, cost, size, and/or the use of consumables that need to be stored or supplied. Lynntech has developed a technology that is based on an innovative, environmentally friendly electrochemical process for the effective removal of ammonium ions. The process does not use consumables except for oxygen gas from air, and does not generate a secondary waste. By controlling operational conditions, the ammonium ions may be transformed to nitrogen gas and/or to nitrate ions. Other advantages of the process include: it is energy efficient, operates at room temperature, and is microgravity compatible. Grant numbers: NAS9 00013. PMID- 11725783 TI - A new photocatalytic reactor for trace contaminant control: a water polishing system. AB - In spacecraft water recovery systems there is a need to develop a postprocessor water polishing system to remove organic impurities to levels below 250 micrograms/L (ppb) with a minimum use of expendables. This article addresses the development of a photocatalytic process as a postprocessor water polishing system that is microgravity compatible, operates at room temperature, and requires only a minimal use of both oxygen gas (or air) and electrical power for low energy UV A (315-400 nm) lamps. In the photocatalytic process, organic contaminants are degraded to benign end products on semiconductor surfaces, usually TiO2. Some challenging issues related to the use of TiO2 for the degradation of organic contaminants have been addressed. These include: i) efficient and stable catalytic material; ii) immobilization of the catalyst to produce a high surface area material that can be used in packed-bed reactors, iii) effective light penetration, iv) effective, microgravity-compatible, oxidant delivery; v) reduced pressure drop, and vi) minimum retention time. The research and development performed on this photocatalytic process is presented in detail. Grant numbers: NAS9-97182. PMID- 11725784 TI - An optimal control strategy for crop growth in advanced life support systems. AB - A feedback control method for regulating crop growth in advanced life support systems is presented. Two models for crop growth are considered, one developed by the agricultural industry and used by the Ames Research Center, and a mechanistic model, termed the Energy Cascade model. Proportional and pointwise-optimal control laws are applied to both models using wheat as the crop and light intensity as the control input. The control is particularly sensitive to errors in measurement of crop dry mass. However, it is shown that the proposed approach is a potentially viable way of controlling crop growth as it compensates for model errors and problems associated with applying the desired control input due to environmental disturbances. Grant numbers: NGT5-50229. PMID- 11725785 TI - Development of a microgravity-compatible reagentless organic acid and alcohol monitor (OAAM). AB - The development of a microgravity-compatible analyzer capable of quantifying organic acids in water is described. The analyzer employs "reagentless" solid phase acidification to convert organic acids to the volatile form followed by membrane separation and specific conductance detection to determine organic acids at concentrations between 0.005 and 40 mg/L. In the future, this technology will be extended to the detection of alcohols, which will be oxidized to form the corresponding organic acid and then determined using the same processes. An immobilized enzyme biocatalyst, alcohol oxidase, oxidizes alcohols to form an aldehyde. Oxidation to the corresponding organic acid is then completed over a heterogeneous catalyst. The combined organic acid and alcohol monitor (OAAM) will be utilized to determine levels of both analyte classes at various points within the water recovery system (WRS) baselined for the International Space Station (ISS). These data will improve water quality through enhanced process control, while allowing early diagnosis of potential problems. Grant numbers: NAG9-1081. PMID- 11725786 TI - Requirements for testing human blood donors for evidence of infection due to communicable disease agents. Final rule. AB - The Food and Drug Administration (FDA) is revising the general biological product standards applicable to human blood and blood components by updating the hepatitis B virus (HBV) and human immunodeficiency virus (HIV) testing requirements, by adding testing requirements for hepatitis C virus (HCV), human T lymphotropic virus (HTLV), and by adding requirements for supplemental (i.e., additional, more specific) testing approved for such use by FDA when a donation is found to be reactive for any of the required screening tests for evidence of infection due to communicable disease agents. The agency also is requiring manufacturers of certain test kits to use reference panels, when available, to verify the acceptable sensitivity and specificity of each lot. This final rule is intended to help protect the safety and ensure the quality of the Nation's blood supply, to enhance the safety of medical devices containing blood or blood components, to provide FDA with clear enforcement authority, and to promote consistency in the industry. Elsewhere in this issue of the Federal Register, FDA is publishing a rule requiring blood and plasma establishments to notify donors, including autologous donors, whenever the donor is deferred or determined not to be suitable for current or future donations of blood and blood components. PMID- 11725787 TI - General requirements for blood, blood components, and blood derivatives; donor notification. Final rule. AB - The Food and Drug Administration (FDA) is amending the biologics regulations to require blood and plasma establishments to notify donors, including autologous donors, whenever the donor is deferred or determined not to be suitable for current or future donations of blood and blood components. A donor is deferred based on results of tests for communicable disease agents or determined not to be suitable for donation based on failure to satisfy suitability criteria. Blood and plasma establishments also are required to notify the referring physician of an autologous donor when the autologous donor is deferred based on tests for evidence of infection with a communicable disease agent(s). A standard operating procedure (SOP) and recordkeeping also are required. This final rule is intended to help protect public health and to promote consistency in the industry. Elsewhere in this issue of the Federal Register, FDA is publishing a final rule on the requirements for testing human blood donors for evidence of infection due to communicable disease agents. PMID- 11725788 TI - Uses of error: genetic counselling. PMID- 11725789 TI - High relative humidity increases yield, harvest index, flowering, and gynophore growth of hydroponically grown peanut plants. AB - Growth chamber experiments were conducted to study the physiological and growth response of peanut (Arachis hypogaea L.) to 50% and 85% relative humidity (RH). The objective was to determine the effects of RH on pod and seed yield, harvest index, and flowering of peanut grown by the nutrient film technique (NFT). 'Georgia Red' peanut plants (14 days old) were planted into growth channels (0.15 x 0.15 x 1.2 m). Plants were spaced 25 cm apart with 15 cm between channels. A modified half-Hoagland solution with an additional 2 mM Ca was used. Solution pH was maintained between 6.4 and 6.7, and electrical conductivity (EC) ranged between 1100 and 1200 microS cm-1. Temperature regimes of 28/22 degrees C were maintained during the light/dark periods (12 hours each) with photosynthetic photon flux (PPF) at canopy level of 500 micromoles-m-2s-1. Foliage and pod fresh and dry weights, total seed yield, harvest index (HI), and seed maturity were greater at high than at low RH. Plants grown at 85% RH had greater total and individual leaflet area and stomatal conductance, flowered 3 days earlier and had a greater number of flowers reaching anthesis. Gynophores grew more rapidly at 85% than at 50% RH. PMID- 11725790 TI - Growth, pod, and seed yield, and gas exchange of hydroponically grown peanut in response to CO2 enrichment. AB - The effects of elevated CO2 on growth, pod, and seed yield, and gas exchange of 'Georgia Red' peanut (Arachis hypogaea L.) were evaluated under controlled environmental conditions. Plants were exposed to concentrations of 400 (ambient), 800, and 1200 micromoles mol-1 CO2 in reach-in growth chambers. Foliage fresh and dry weights increased with increased CO2 up to 800 micromoles mol-1, but declined at 1200 micromoles mol-1. The number and the fresh and dry weights of pods also increased with increasing CO2 concentration. However, the yield of immature pods was not significantly influenced by increased CO2. Total seed yield increased 33% from ambient to 800 micromoles mol-1 CO2, and 4% from 800 to 1200 micromoles mol 1 CO2. Harvest index increased with increasing CO2. Branch length increased while specific leaf area decreased linearly as CO2 increased from ambient to 1200 micromoles mol-1. Net photosynthetic rate was highest among plants grown at 800 micromoles mol-1. Stomatal conductance decreased with increased CO2. Carboxylation efficiency was similar among plants grown at 400 and 800 micromoles mol-1 and decreased at 1200 micromoles mol-1 CO2. These results suggest that CO2 enrichment from 400 to 800 micromoles mol-1 had positive effects on peanut growth and yield, but above 800 micromoles mol-1 enrichment seed yield increased only marginally. PMID- 11725791 TI - Generic drugs: the stalling game. AB - Sweetheart deals and patent extensions keep lower-cost generic drugs from consumers. PMID- 11725792 TI - Ceratopteris richardii: a productive model for revealing secrets of signaling and development. AB - Ceratopteris richardii is an aquatic fern grown in tropical and subtropical regions of the world. It is proven to be a productive model system for studies in the genetics, biochemistry, and cell biology of basic biologic processes that occur in early gametophytic development. It provides several advantages to biologists, especially those interested in gravitational biology, polarity development, and in the genetics of sexual development. It is easy to culture, has a relatively short life cycle, and offers an array of attractive features that facilitate genetic studies. The germination and early development of large populations of genetically identical spores are easy to synchronize, and both the direction of polarity development and cell-level gravity responses can be measured and readily manipulated within the first 24 h of spore development. Although there is no reliable transformation system available yet in Ceratopteris, recent studies suggest that the technique of RNA interference can be used to block translation of specific genes in a related fern, Marsilea, and current studies will soon reveal the applicability of this approach, as well as of other transformation approaches, in Ceratopteris. A recently completed expressed sequence tag (EST) sequencing project makes available the partial sequence of more than 2000 cDNAs, representing a significant percentage of the genes being expressed during the first 24 h of spore germination, when many developmentally interesting processes are occurring. A microarray of these ESTs is being constructed, so especially for those scientists interested in basic cellular phenomena that occur early in spore germination, the availability of the ESTs and of the microarray will make Ceratopteris an even more attractive model system. PMID- 11725793 TI - Vaccines: an issue of trust. Misinformation and government foot-dragging are fanning fears. PMID- 11725794 TI - Employee Retirement Income Security Act of 1974; rules and regulations for administration and enforcement; claims procedure. Final regulation; delay of applicability date. AB - This action delays for at least six months and not more than one year the applicability date for the regulation governing minimum requirements for benefit claims procedures of group health plans covered by Title I of the Employee Retirement Income Security Act. As published on November 21, 2000, the benefit claims procedure would be applicable to claims filed on or after January 1, 2002. The current action amends the regulation so that it will apply to group health claims filed on or after the first day of the first plan year beginning on or after July 1, 2002, but in no event later than January 1, 2003. This action provides a limited additional period within which group health plan sponsors, administrators, and service providers can bring their claims processing systems into compliance with the new requirements. A postponement of the applicability date with respect to group health claims will allow a more orderly transition to the new standards and will avoid the confusion and additional expense that would be caused if certain pending Congressional bills are enacted before or soon after the original applicability date. This action does not apply to pension plans or plans providing disability or welfare benefits (other than group health). For these plans, the regulation will continue to be applicable to claims filed on or after January 1, 2002. PMID- 11725795 TI - Neutron yields from 435 MeV/nucleon Nb stopping in Nb and 272 MeV/nucleon Nb stopping in Nb and Al. AB - Neutron fluences were measured from 435 MeV/nucleon Nb ions stopping in a Nb target and 272 MeV/nucleon Nb ions stopping in targets of Nb and Al for neutrons above 20 MeV and at laboratory angles between 3 degrees and 80 degrees. The resultant spectra were integrated over angles to produce neutron energy distributions and over energy to produce neutron angular distributions. The total neutron yields for each system were obtained by integrating over the angular distributions. The angular distributions from all three systems are peaked forward, and the energy distributions from all three systems show an appreciable yield of neutrons with velocities greater than the beam velocity. Comparison of the total neutron yields from the two Nb + Nb systems suggests that the average neutron multiplicity decreases with decreasing projectile energy. Comparison of the total yields from the two 272 MeV/nucleon systems suggests that the total yields show the same dependence on projectile and target mass number as do total inclusive neutron cross sections. The data are compared with Boltzmann-Uehling Uhlenbeck model calculations. PMID- 11725796 TI - Computing in comfort. A well-designed workstation can prevent injuries and make you more productive. PMID- 11725797 TI - Milk: got proof? We assess the evidence behind the dairy industry's ads and critics' claims. PMID- 11725798 TI - Rules of practice: medical opinions from the Veterans Health Administration. Interim final rule with request for comments. AB - This document amends the Department of Veterans Affairs' (VA) Appeals Regulations to clarify that the Board of Veterans' Appeals (Board) may obtain medical opinions from health care professionals in VA's Veterans Health Administration. PMID- 11725799 TI - Statement of organization and functions. Final rule. AB - The Consumer Product Safety Commission is amending its statement of organization and functions to reflect the transfer of the National Injury Information Clearinghouse from the Office of Information Services to the Directorate for Epidemiology. PMID- 11725800 TI - Medical devices; exemption from premarket notification requirements; class I devices; technical amendment. Final rule; technical amendment. AB - The Food and Drug Administration (FDA) is amending the language in its medical device classification regulations for class I devices for consistency, to include in sections where it was not present, a specific reference to the limitations on exemptions from premarket notification requirements for each generic device classified. The specific reference language was included when some class I generic devices were first exempted under provisions of the Food and Drug Administration Modernization Act of 1997 (FDAMA). These amendments will provide the same reference for devices that were exempted before that time. The language is intended to conveniently provide the reference, and make the sections clear and easy to read. The status of the devices is not being changed. PMID- 11725801 TI - Evidence of root zone hypoxia in Brassica rapa L. grown in microgravity. AB - A series of experiments was conducted aboard the U.S. space shuttle and the Mir space station to evaluate microgravity-induced root zone hypoxia in rapid-cycling Brassica (Brassica rapa L.), using both root and foliar indicators of low-oxygen stress to the root zone. Root systems from two groups of plants 15 and 30 d after planting, grown in a phenolic foam nutrient delivery system on the shuttle (STS 87), were harvested and fixed for microscopy or frozen for enzyme assays immediately postflight or following a ground-based control. Activities of fermentative enzymes were measured as indicators of root zone hypoxia and metabolism. Following 16 d of microgravity, ADH (alcohol dehydrogenase) activity was increased in the spaceflight roots 47% and 475% in the 15-d-old and 30-d-old plants, respectively, relative to the ground control. Cytochemical localization showed ADH activity in only the root tips of the space-grown plants. Shoots from plants that were grown from seed in flight in a particulate medium on the Mir station were harvested at 13 d after planting and quick-frozen and stored in flight in a gaseous nitrogen freezer or chemically fixed in flight for subsequent microscopy. When compared to material from a high-fidelity ground control, concentrations of shoot sucrose and total soluble carbohydrate were significantly greater in the spaceflight treatment according to enzymatic carbohydrate analysis. Stereological analysis of micrographs of sections from leaf and cotyledon tissue fixed in flight and compared with ground controls indicated no changes in the volume of protoplast, cell wall, and intercellular space in parenchyma cells. Within the protoplasm, the volume occupied by starch was threefold higher in the spaceflight than in the ground control, with a concomitant decrease in vacuolar volume in the spaceflight treatment. Both induction of fermentative enzyme activity in roots and accumulation of carbohydrates in foliage have been repeatedly shown to occur in response to root zone oxygen deprivation. These results indicate that root zone hypoxia is a persistent challenge in spaceflight plant growth experiments and may be caused by microgravity-induced changes in fluid and gas distribution. PMID- 11725802 TI - Water recovery in space. AB - In the absence of recycling, water represents over 90% of the life-support consumables for a manned spacecraft. In addition, over 90% of the waste water generated can be classified as moderately or slightly contaminated (e.g. shower water, condensate from the air-conditioning system, etc.). The ability to recover potable water from moderately contaminated waste water hence enables significant savings to be made in resupply costs. A development model of such a water recovery system, based on membrane technology has been produced and tested using "real waste water" based on used shower water Results indicate some 95% recovery of potable water meeting ESA standards, with total elimination of microbial contaminants such as bacteria, spores and viruses. PMID- 11725803 TI - Montgomery GI Bill--active duty. Final rule. AB - This document amends the educational assistance regulations of the Department of Veterans Affairs (VA). The amendments reflect statutory changes contained in the Veterans Millennium Health Care and Benefits Act of 1999 and statutory interpretations. This document also makes changes for the purpose of clarification. PMID- 11725804 TI - Medicare and Medicaid Programs; change of agency name: technical amendments. Final rule with comment period. AB - In accordance with the name change of the Health Care Financing Administration to the "Centers for Medicare & Medicaid Services," this technical regulation revises all references to "Health Care Financing Administration" and "HCFA" in chapters I, IV and V of title 42 and subtitle A and chapters II and III of title 45 of the Code of Federal Regulations. This regulation also makes conforming changes to the general definitions sections. PMID- 11725805 TI - The Americans with Disabilities Act as risk regulation. AB - Much disability based discrimination occurs because of fears that hiring or serving people with disabilities will pose a safety risk. Disability rights laws such as the ADA strictly regulate such risk-motivated discrimination. Many disability rights advocates and academic defenders of the ADA laud such laws as applying a "scientific" approach to risk rather than the "irrational" approach generally adopted by the public at large. That position is doubly strange: It stands in remarkable tension with disability rights advocates' general suspicion of "experts," and applies a technocratic approach to risk regulation-an approach that usually has politically conservative implications-to achieve the distinctly nonconservative goal of promoting the full integration of people with disabilities into our nation's economic and civic life. In this essay, Professor Bagenstos uses the problem of risk-motivated disability discrimination as a lens through which to examine the politics of risk regulation scholarship. He argues that the easy association of technocratic approaches with political conservatism and of democratic approaches with political liberalism or progressivism-ignores the complex ways in which technocratic and democratic institutions may serve or disserve particular political interests. PMID- 11725806 TI - Antenatal corticosteroids revisited: repeat courses. AB - OBJECTIVE: To provide health care providers, patients, and the general public with a responsible assessment of currently available data regarding the benefits and risks of repeat courses of antenatal corticosteroids. PARTICIPANTS: A non Federal, non-advocate, 16-member panel representing the fields of obstetrics and gynecology, pediatrics, maternal and fetal medicine, neonatology, medical ethics, community health, pharmacology, psychology, and reproductive biology. In addition, 13 experts in these same fields presented data to the panel and to a conference audience of approximately 200. EVIDENCE: The literature was searched using MEDLINE and an extensive bibliography of references was provided to the panel. Experts prepared abstracts with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed their conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. The draft statement was made available on the World Wide Web immediately following its release at the conference and was updated with the panel's final revisions. CONCLUSIONS: The collective international data continue to support unequivocally the use and efficacy of a single course of antenatal corticosteroids using the dosage and interval of administration specified in the 1994 Consensus Development Conference report. The current benefit and risk data are insufficient to support routine use of repeat or rescue courses of antenatal corticosteroids in clinical practice. Clinical trials are in progress to assess potential benefits and risks of various regimens of repeat courses. Until data establish a favorable benefit-to-risk ratio, repeat courses of antenatal corticosteroids, including rescue therapy, should be reserved for patients enrolled in clinical trials. PMID- 11725807 TI - Avoiding pitfalls in chronic disease quality measurement: a case for the next generation of technical quality measures. AB - The true utility of quality measurement lies in its ability to inspire quality improvement, with resultant enhancements in the processes and outcomes of care. Because quality measurement is expensive, it is difficult to justify using measures that are not likely to lead to important improvements in health. Many current measures of chronic disease technical quality, however, have one or more pitfalls that prevent them from motivating quality improvement reactions. These pitfalls include that: (1) measured processes of care lack strong links to outcomes; (2) actionable processes of care are not measured; (3) measures do not target those at highest risk; (4) measures do not allow for patient exceptions; and (5) intermediate outcome measures are not severity adjusted. To exemplify recent advancements and current pitfalls in chronic disease quality measurement, we examine the evolution of quality measures for diabetes mellitus and discuss the limitations of many currently used diabetes mellitus care measures. We then propose more clinically meaningful "tightly linked" measures that examine clinical processes directly linked to outcomes, target populations with specific diagnoses or intermediate disease outcomes that contribute to risk for poor downstream health outcomes, and explicitly incorporate exceptions. We believe that using more tightly linked measures in quality assessment will identify important quality of care problems and is more likely to produce improved outcomes for those with chronic diseases. PMID- 11725808 TI - Effects of an HMO hospitalist program on inpatient utilization. AB - OBJECTIVE: To assess the impact of a health maintenance organization (HMO) hospitalist program on inpatient utilization. PATIENTS: The study sample consisted of patients admitted to the hospital for a routine, uncomplicated acute surgical or medical diagnosis included under Milliman and Robertson's Optimal Recovery Guidelines (ORGs). Evaluation involved comparison of 2 physician management groups: inpatients managed by staff primary care physicians (PCPs) (study group) and inpatients managed by network PCPs (comparison group). STUDY DESIGN: Data before and after introduction of the hospitalist program were available. From the same period, data were available from patients of network PCPs who managed their own inpatient stays (nonhospitalist comparison sample). Three outcomes were used to represent inpatient utilization: (1) number of inpatient stays meeting ORG goals; (2) number of inpatient stays not meeting ORG goals that had > or =1 medically necessary days; and (3) number of inpatient stays not meeting ORG goals that had > or =1 medically unnecessary days. RESULTS: Multiple logistic regression results indicated that inpatient stays were more likely to be within ORG utilization goals when managed by hospitalists vs nonhospitalists (P < .05). Introduction of the hospitalist program reduced the number of stays with unnecessary days among staff inpatients. There was an increase in stays with unnecessary days in the comparison group, ie, inpatients managed by network physicians. CONCLUSIONS: Full-time hospitalists are efficient managers of HMO inpatients. The ORGs for acute, uncomplicated diagnoses provided useful hospital utilization measures that captured inpatient management by hospitalists and PCPs. PMID- 11725809 TI - Managed care in the doctor's office: has the revolution stalled? AB - OBJECTIVE: To assess trends in the involvement of US physicians with managed care. STUDY DESIGN: Comparison of data from 2 consecutive rounds of a national survey. METHODS: Longitudinal data were obtained from the 1996/1997 (n = 12,528) and the 1998/1999 (n = 12,304) rounds of the Community Tracking Study (CTS) Physician Survey, a large, ongoing nationally representative survey of US physicians involved in patient care. Indicators used to assess involvement with managed care included global measures of managed care participation, risk contracting, exposure to financial incentives, and impact of care management tools. Changes in these measures over the 2 study periods are reported. Analyses were conducted for all physicians, as well as for primary care physicians (PCPs) and specialists separately. RESULTS: The percentage of practice revenue derived from managed care increased only modestly over the study period (from 42% to 45%). Mean numbers of managed care contracts per physician increased minimally (from 12 to 13). Trends in acceptance of capitation and exposure to financial incentives remained stable over the study period. Among PCPs, employment in staff/group health maintenance organizations declined slightly, whereas gatekeeping function increased modestly. Among care management tools, only treatment guidelines had a significantly increased impact on medical practice, primarily among PCPs (from 46% to 52%; P < .001). CONCLUSIONS: Many aspects of managed care leveled off between 1996 and 1999 in ways not accurately reflected by plan enrollment patterns. This "flattening of the curve" trend appears to hold generally across multiple measures. A stalling of the managed care "revolution," if it is sustained, may portend future escalation in healthcare costs. PMID- 11725810 TI - A new measure of the impact of managed care on healthcare markets. AB - BACKGROUND: Most studies of managed care impact have used health maintenance organization (HMO) penetration or index of competition as the marker of managed care impact. However, little empirical evidence has been found to support the validity of these or other measures in current use. In addition, as managed care evolves to forms other than HMOs and managed care penetration in large metropolitan areas approaches 100% of commercially insured patients, the utility of the most commonly used measure, HMO penetration, will decrease still further. OBJECTIVES: To provide a preliminary analysis of the use of premiums as a measure of market impact of managed care. STUDY DESIGN: Retrospective analysis (quartile, correlation, multiple-variable linear regression) of publicly available datasets. METHODS: Labor market-adjusted HMO premiums from 3 publicly available sources, for the 56 largest metropolitan areas in the United States, were compared with penetration and index of competition as predictors of the dependent market variable, hospital bed-days per 1000 population. RESULTS: Health maintenance organization premiums in the Federal Employees Health Benefits Program emerged as the best predictor of HMO market impact. Average HMO premiums reported in the Interstudy database and for the Medicare+Choice program also outperformed penetration or index of competition in relating to several commonly available markers of competition such as bed-days per 1000. CONCLUSIONS: Premiums charged by HMOs are a useful measure of the impact of managed care on healthcare markets in large metropolitan areas. PMID- 11725811 TI - Group practice strategies to manage pharmaceutical cost in an HMO network. AB - OBJECTIVE: To evaluate the prevalence of various pharmaceutical cost management strategies used by group practices within a managed care network and their relationship to drug costs among enrollees. STRATEGIES STUDIED: Care management (gatekeeping, practice profiling, practice guidelines, case management), techniques for maintaining clinic medication records, and policies regulating physician interaction with pharmaceutical sales representatives (PSRs). STUDY DESIGN: Cross-sectional survey of primary care group practice organizations (n = 103) affiliated with Blue Cross Blue Shield of Minnesota in early 1996. METHODS: Multivariate linear regression analysis was performed on corresponding claims data for members continuously enrolled in these practices from January 1 to December 31, 1995 (n = 76,387), using the patient as the unit of analysis. RESULTS: Substantial variation in strategy prevalence was observed; this variation was thought to influence pharmaceutical costs. Seventy-six percent of practices had medication lists in outpatient medical records, 53% had policies limiting pharmaceutical detailing, and 44% had patients assigned to primary care gatekeepers; however, only 10% used outpatient nurse case managers. Use of outpatient nurse case managers (P < .010), primary care physician gatekeeping (P < .002), policies to control pharmaceutical detailing (P < .001), and medication lists and outpatient charts (P < .001) was found to be independently associated with lower pharmaceutical expenditures. Significant colinearity was found between group size and the strategies studied. CONCLUSIONS: Significantly lower pharmaceutical costs per member per year were observed in the groups reporting primary care gatekeeping, outpatient medication records, outpatient case managers, and policies regarding physician interactions with PSRs. PMID- 11725812 TI - Effect of prescription benefit changes on medical care utilization in a Medicare HMO population. AB - OBJECTIVE: To examine the impact of 2 cost-containment efforts in prescription benefits in successive years that included changes in copayment and coverage levels, expanded generic coverage, and brand name prescription drug limit-of coverage in a Medicare health maintenance organization (HMO). The benefit changes included moving to a drug benefit with increased total coverage and higher copayments in the first year (1998) and to one with brand name limit-of-coverage and unlimited generic availability in the second year (1999). STUDY DESIGN: A repeated-measures analytical design with enrollee follow-up before and after introduction of the 2 policies. PATIENTS AND METHODS: A cohort of 2411 older adults continuously enrolled in a Medicare HMO since 1998 was followed up for 1 year pre-post for healthcare service utilization and costs; 259 patients enrolled since 1997 were available to test the effects of the first policy change. RESULTS: Bivariate and multivariate analyses found a significant decrease of 27% in prescription costs, a 4% decrease in physician visits, and a 6% decrease in total costs associated with the change in prescription benefit in the second year (1999). The policy change in the first year (1998) resulted in a 29% increase in prescription costs and 38% increased total costs for the HMO. CONCLUSIONS: Introduction of a prescription benefit that included substantial brand name limit of-coverage and generic drug coverage expansion was associated with significantly reduced prescription costs. In addition, this change did not seem to increase nonprescription-related healthcare service use in the population. PMID- 11725813 TI - Who will pay for the value of hospitalists? PMID- 11725814 TI - Effects of single and repeated administration of 1,2,3,4-tetrahydroisoquinoline analogs on the binding of [11C]raclopride to dopamine D2 receptors in the mouse brain. AB - We investigated the effects of intraperitoneal injection of 1,2,3,4 tetrahydroisoquinoline (TIQ) analogs and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) on the binding of [11C]raclopride to striatal dopamine D2 receptors in mice. The binding of [11C]raclopride, but not of [11C]N methylspiperone or [11C]nemonapride with higher affinity, to the receptors was significantly decreased immediately after TIQ injection. Neither a dopamine transporter blocker induced such effect nor TIQ affected the dopamine transporter radioligand binding. Among the compounds investigated, including parkinsonism inducing TIQ and (R/S)-1-benzyl-TIQ, parkinsonism-preventing (R)- and (S)-1 methyl-TIQ, and probable N-methylated metabolites of TIQ and 1-methyl-TIQ, TIQ and (S)-1-methyl-TIQ had the strongest effect on the binding of [11C]raclopride, and N-methylated derivatives showed less of an effect than the respective parent compounds. The decrease in the binding of [11C]raclopride continued for 7 hours and was followed by an increase until 10 days after the single and subchronic administration of TIQ. These findings suggest that TIQ analogs profoundly stimulated dopamine release which resulted in the competitive inhibition of the binding of [11C]raclopride to dopamine D2 receptors, but did not induce degeneration of the receptors. PMID- 11725815 TI - Stimulation of the regrowth of MPP+-damaged dopaminergic fibers by the treatment of mesencephalic cultures with basigin. AB - Basigin (Bsg) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily and widely expressed in the central nervous system. To elucidate functional role of Bsg in the central nervous system, the effects of its glutathione-S-transferase (GST) fusion protein on the number and neurite outgrowth of cultured rat mesencephalic dopaminergic neurons were measured. The fusion protein was not able to promote the survival and neurite outgrowth of tyrosine hydroxylase (TH)-positive neurons under serum-free condition. However, the treatment of 1-methyl-4-phenylpyridinium (MPP+)-exposed cultures with the fusion protein resulted in stimulation of the regrowth of damaged TH-positive fibers. Basic fibroblast growth factor (bFGF) also stimulated the regrowth of neurites in damaged neurons. These results indicate that Bsg may play an important role in the regrowth of damaged dopaminergic fibers. PMID- 11725816 TI - Parkinson's disease is associated with oxidative stress: comparison of peripheral antioxidant profiles in living Parkinson's, Alzheimer's and vascular dementia patients. AB - Antioxidant profiles in Parkinson's disease (PD; n = 15), dementias of Alzheimer's type (DAT; 18) and Vascular (VD; 15), and control subjects (C; 14) were studied. Cu-Zn superoxide dismutase (SOD), catalase (CAT), glutathione system (GLU) and thiobarbituric acid reactive substances (TBARS) were measured in erythrocytes; antioxidant capacity (TRAP) in plasma. Biochemical variables were analyzed simultaneously using multi-variate and non-parametric methods. Clinical diagnostic resulted associated with the main source of variability in antioxidant variables (Kruskal-Wallis: H = 32.58, p = 0.000001). Comparison of PD and C resulted highly significant (z = 4.47, p = 0.000047), demonstrating an association between oxidative stress and PD. SOD and TBARS were significantly higher in pathological groups against C (p = 0.0000001, p = 0.051); TRAP resulted lower (p = 0.00015). Discriminant functions constructed using biochemical variables separated pathological groups (93% success) from C, and DAT (88.9%) from VD (73.3%); but not PD from DAT or VD. Antioxidant profiles of PD patients showed characteristics overlapping with DAT (60%) and with VD (40%), suggesting biochemical similarities between them. PMID- 11725817 TI - Correlation of nicotinic receptor binding with clinical and neuropathological changes in Alzheimer's disease and dementia with Lewy bodies. AB - We investigated the relationship between the loss of nicotinic acetylcholine receptors (nAChR) and the cognitive decline or neuropathological changes seen in Alzheimer's Disease (AD) and dementia with Lewy bodies (DLB). Midfrontal (MF) cortex of 31 AD, 24 DLB and 11 non-demented controls was examined. Total plaque (TP), neuritic plaque (NP) and neurofibrillary tangle (NFT) counts were obtained. NAChR binding was assayed using 3H-epibatidine [3H-EPI]. Last Blessed Information Memory-Concentration scores (BIMC), Mini-Mental State Examination (MMSE), Mattis Dementia Rating Scale (DRS) scores were collected. There were no correlations between 3H-EPI binding and TP, NP, NFTs counts in either AD or DLB. Last BIMC, MMSE, DRS scores did not correlate with 3H-EPI binding in AD or DLB. Thus, decline in cognitive function does not correlate with loss of nAChR in DLB or AD at the end of life suggesting that later in these diseases, loss of nAChR binding is not a reliable marker of cognitive function in AD or DLB. Loss of nAChR activity does not appear to be related to plaques or NFTs in AD or DLB. PMID- 11725818 TI - Neither the butyrylcholinesterase K variant nor transferrin C2 variant confers a risk for Alzheimer's disease in Koreans. AB - To investigate the possible involvement of the butyrylcholinesterase (BCHE) K variant and transferrin (TF) C2 variant in the manifestation of Alzheimer's disease (AD), we analyzed the BCHE, TF and apolipoprotein E (APOE) genotypes of 164 sporadic AD patients and 239 normal elderly controls. The frequencies of the BCHE K and TF C2 did not differ between the AD patients and controls (P > 0.1). The occurrence of the APOE epsilon4 did not influence the distribution of the BCHE K and TF C2 variants (P > 0.1). No linkage disequilibrium between the BCHE K and TF C2 was observed either in both the AD patients and controls (P > 0.1). In conclusion, neither the BCHE K nor the TF C2 confers a risk for AD. PMID- 11725819 TI - EEG changes during long-term treatment with donepezil in Alzheimer's disease patients. AB - In this pilot study, we examined the long-term treatment effect of donepezil on the quantitative EEG (qEEG) in 12 Alzheimer's disease patients. The qEEGs of the mean absolute and relative amplitudes of betal, alpha, theta and delta activities were obtained at baseline and during donepezil treatment. Comparisons of awake qEEG prior to and during treatment were performed using a 2-way analysis of variance (ANOVA) with repeated measures. In patients with mild dementia (n = 5), the qEEG analysis showed a significant reduction of the mean absolute theta activity (p = 0.05) by donepezil, particularly in frontal and temporo-parietal areas. In patients with moderate/severe dementia (n = 7), a significant decrease in the mean absolute beta 1 activity (p = 0.02), particularly in the frontal and occipital areas may be attributed to disease progression which was not counteracted by the long-term treatment. The differences in qEEG in patients with different stages of dementia under donepezil treatment may be related to different compensatory capacities due to structural and functional brain disturbances. PMID- 11725820 TI - Association analysis of HTR6 and HTR2A polymorphisms in sporadic Alzheimer's disease. AB - In order to identify gene variants related to the serotonergic neurotransmitter system that possibly represent a hereditary risk factor for sporadic Alzheimer's disease (AD), patients suffering from AD and non-demented psychiatric inpatients without symptoms of dementia were genotyped for polymorphisms of HTR6 (267C/T) and HTR2A (-1438G/A). Although there was a tendency toward an increased number of the genotype TT of the 5-HT6 receptor polymorphism in AD patients when compared to controls (2.8% vs. 1.3%), neither this nor the 5-HT2A promoter polymorphism showed significant differences in their genotypic or allelic distribution among patients and controls. These polymorphisms probably do not represent major genetic risk factors of AD. However, further studies including other genetic variants of the serotonergic neurotransmitter system are needed in order to elucidate their role in AD. PMID- 11725821 TI - A behavioural pattern analysis of hypoglutamatergic mice--effects of four different antipsychotic agents. AB - In a hypoglutamatergic rodent model, we have observed certain behaviours that might have relevance for the cognitive impairments seen in autism and schizophrenia. Thus, hypoglutamatergic mice show defective habituation, impaired attention, a meagre behavioural repertoire and a general behavioural primitivization. The aim of the present study was to characterise and quantify changes in movement pattern in mice rendered hypoglutamatergic by means of MK-801 treatment, using an automated video tracking system. Further, the effects of four different antipsychotic drugs, the classical neuroleptic haloperidol, the atypical antipsychotic clozapine, the DA D2/5-HT2A antagonist risperidone and the selective 5-HT2A-receptor antagonist M100907, were compared with respect to effects on NMDA antagonist-induced movement pattern alterations. We found that each receptor antagonist had a unique effect on the MK-801-induced behavioural primitivization. Haloperidol was unable to affect the monotonous behaviour induced by MK-801, while risperidone, clozapine and M100907 produced movement patterns of high intricacy. PMID- 11725822 TI - Antidepressants do not modulate estrogen receptor alpha-mediated gene expression. AB - The estrogen receptor alpha has been shown to be activated both in a ligand dependent and in a ligand independent fashion. We investigated whether antidepressants may directly activate the estrogen receptor alpha or enhance ligand-dependent or ligand-independent activation of the estrogen receptor alpha. Whereas both ligand-dependent and ligand-independent activation of the estrogen receptor alpha could be demonstrated in transient transfection studies, antidepressants neither directly activated the estrogen receptor alpha nor did they enhance ligand-dependent or ligand-independent activation. Thus, there are differences between the glucocorticoid receptor and the estrogen receptor alpha with regard to ligand-independent activation and action of antidepressants. PMID- 11725823 TI - Prefrontal blood flow dysregulation in drug naive ADHD children without structural abnormalities. AB - Recent studies suggest a role for prefrontal cortex abnormalities in the pathogenesis of attention deficit/hyperactivity disorder (ADHD). We evaluated young drug-naive ADHD outpatients without MRI structural abnormalities to detect prefrontal cortex regional cerebral blood flow (rCBF) functional dysregulation; correlation between age and rCBF; and correlation between symptoms profile and rCBF. Functional brain activities (i.e. rCBF), neuropsychological attention performance and symptom profile were evaluated respectively by single photon emission computerized tomography (SPECT) scan, Stroop Test and the Child Attention Problem Rating Scale. There was a decreased rCBF in the left dorso lateral prefrontal cortex (DLPFC) compared to the right DLPFC of the subjects. In addition, there were positive correlations between age and relative rCBFs of the dorsolateral and orbital prefrontal cortex, and negative correlations between age and absolute rCBFs of the dorsolateral and orbital prefrontal cortex. Finally, higher levels of right relative rCBF and lower levels of left relative rCBF were predictors of higher severity of clinical symptom expression and neuropsychological attention impairment. The results of this study highlight the role of the DLPFC blood flow impairment in the pathogenesis of ADHD even in young subjects without structural abnormalities. PMID- 11725824 TI - Comparison of the efficacy of 2 mg versus 5 mg tropisetron in the management of post-operative nausea and vomiting. AB - A randomized, double-blind, placebo-controlled study was carried out to investigate the efficacy of 2 mg versus 5 mg tropisetron in the prevention of post-operative nausea and vomiting. Patients between 22 and 64 years old who were undergoing surgery under general anaesthesia and who had been classified according to the American Society of Anesthesiologists criteria with physical status I or II were included in this trial. Sixty female patients were recruited and divided into three groups (n = 20 in each group). Patients in group 1 received saline solution as control, whereas in groups 2 and 3, 2 mg or 5 mg tropisetron, respectively, was administered intravenously as a single dose during the induction of anaesthesia. The number of patients vomiting within the first 2 h following the operation was significantly less in groups 2 and 3 compared with that in group 1 (one of 20 for each of the groups 2 and 3 versus 12 of 20 in group 1). Three patients required rescue anti-emetic medication in each of the groups 2 and 3. In conclusion, 2 mg tropisetron appears to be equally as efficacious as 5 mg in preventing post-operative nausea and vomiting. PMID- 11725825 TI - Effect of cisapride on gastric emptying in patients with gastro-oesophageal reflux disease. AB - To investigate the effect of cisapride in gastro-oesophageal reflux disease (GERD), gastric emptying time, measured using scintigraphy, and reflux symptoms were compared before and after cisapride therapy in 30 patients with endoscopically proven GERD. Gastric emptying time was also measured in 20 age matched controls. Patients with GERD were treated with 30 mg cisapride orally three times daily for 7 days. Gastric emptying time was significantly reduced following cisapride therapy in 28 of these patients (71.6 +/- 18.1 min versus 57.9 +/- 13.9 min), although it was still longer than the gastric emptying time of the control group (i.e. 46.2 +/- 8.1 min). In addition, cisapride relieved heartburn, which is a representative symptom of GERD. These findings suggest that cisapride may be useful in treating reflux symptoms and oesophagitis in patients with GERD. PMID- 11725826 TI - Conjugated linoleic acid reduces body fat in healthy exercising humans. AB - This study was designed to investigate the efficacy and tolerability of daily conjugated linoleic acid (CLA) in healthy exercising humans. This was a random double-blind, placebo-controlled study in 20 healthy humans of normal body weight and body mass index less than 25.0 kg/m2, who did standardized physical exercise in a gym for 90 min three times weekly. Participants took either placebo (hydrogel) or CLA 0.6 g [DOSAGE ERROR CORRECTED], three times daily, as two capsules during meals, for 12 weeks. Body fat, measured using near infrared light, was significantly reduced in the CLA group during the study, but not in the placebo group. No effects on body weight were observed. Tolerability was good and similar in the two groups. Compliance, as judged by the number of returned capsules, was more than 80% of the recommended dose for all participants. Thus CLA reduces body fat but not body weight in healthy exercising humans of normal body weight. PMID- 11725827 TI - Detection of point mutation for human growth hormone in patients with anti pituitary antibody positive type 1 diabetes mellitus. AB - We investigated genetic mutations in the coding region of the human growth hormone (hGH) gene in anti-pituitary antibody (APA)-positive patients with type 1 diabetes mellitus (n = 6) or autoimmune thyroid diseases (n = 10) and in APA negative, healthy controls (n = 10). A point mutation in the hGH gene was discovered in two patients with type 1 diabetes mellitus. No mutations were found in the hGH gene in control subjects, patients with autoimmune thyroid diseases (Hashimoto's thyroiditits, Graves' disease) or in the remaining four patients with type 1 diabetes mellitus. The mutation was located in the coding region for the second amino acid in the N-terminal region of hGH. This point mutation was identified in codon 2 in exon 2 of the hGH gene. We successfully developed an allele-specific amplification method for detecting this mutation using the polymerase chain reaction. PMID- 11725828 TI - Bacterial lipopolysaccharide induces transforming growth factor beta and hepatocyte growth factor through toll-like receptor 2 in cultured human colon cancer cells. AB - This study examined, in human cancer lines, the pattern of cytokine production stimulated by lipopolysaccharide (LPS), a major component of outer surface of gram-negative bacteria, and characterized the expression pattern of CD14, cell surface LPS receptor antigen, and toll-like receptors (TLRs), which appear to be key regulators of the innate immune response system. Two colon cancer cell lines (DLD and LoVo), a hepatocellular carcinoma cell line and a myelomonocytic cell line were incubated with LPS for 0-72 h, and transforming growth factor (TGF) beta1 and beta2, hepatocyte growth factor (HGF) and interleukins 6, 8 and 15 were assayed. The only changes induced by incubation with LPS were significant increases in TGFbeta1 production at 12 h, and in HGF production at 72 h, in LPS stimulated DLD cells, and significant increases in TGFbeta2 production after 12 h and in HGF after 72 h in LoVo cells. Using reverse transcriptase-polymerase chain reaction analysis, expression of CD14 and TLR-2 mRNA was detected in DLD and LoVo cells, and expression of TLR-4 mRNA was detected in PLC/PRF/5 and KG-1 cells. These results suggest that LPS induces TGFbeta and HGF production mediated by CD14/TLR-2 in cultured human colon cancer cell lines. PMID- 11725829 TI - Cost-effectiveness of minimally invasive-intervention in aortofemoral revascularization. AB - Minimally invasive surgical procedures have become popular recently because they offer several advantages over conventional operative techniques. A person undergoing a minimally invasive procedure usually experiences less pain, is discharged earlier from hospital, returns to work sooner and has a less obtrusive post-operative scar. Excluding highly technical techniques (e.g. laparoscopic surgery), operations performed using mini-laparotomy are usually more cost effective than conventional procedures, largely because they are less expensive to perform. Our paper investigates cost-effectiveness and other parameters relating to minimally invasive aortofemoral revascularization procedures performed at our clinic. We compared 20 similar cases, half where revascularization was undertaken using mini-laparotomy and half where conventional laparotomy was selected. From our findings we conclude that mini laparotomy is safe and reliable for aortobifemoral bypass procedures and has several advantages over traditional techniques; namely, shorter operating time, earlier resumption of intestinal function, shorter duration of in-patient stay and reduced costs. PMID- 11725830 TI - Autologous peripheral stem-cell transplantation after intensive chemotherapy in a case of CD30 (Ki-1)-positive anaplastic large-cell lymphoma. AB - Simultaneous treatment with peripheral blood stem-cell (PBSC) transplantation and intensive chemotherapy was evaluated in a case of non-Hodgkin's lymphoma (NHL) with poor prognosis. A 59-year-old male diagnosed with a high-grade, anaplastic large-cell (Ki-1) NHL, involving fractures in the left hip, underwent computed tomography and gallium scintigram surveillance. The patient received chemotherapy with epirubicin hydrochloride, cyclophosphamide, vincristine and prednisolone, and the fractured hip bone was repaired following the first course of chemotherapy. After the second and third courses of chemotherapy, PBSCs were harvested and cryopreserved. The patient then received a further course of chemotherapy and PBSC transplantation was conducted using infused cells consisting of 9.63 x 10(6)/kg CD34 cells and 2.24 x 10(5)/kg granulocyte macrophage colony-forming units. Recovery of platelet and white blood cell counts occurred 10 and 8 days, respectively, after PBSC infusion and the patient remains well. PMID- 11725831 TI - Successful treatment of patients with rheumatic disorders and acquired factor VIII inhibitors with cyclophosphamide and prednisolone combination therapy: two case reports. AB - Acquired haemophilia associated with autoimmune disorders can be fatal and has been reported to be refractory to steroid therapy alone. We report two cases of female patients, aged 24 years and 54 years, with acquired haemophilia caused by factor VIII inhibitors. Underlying diseases were systemic lupus erythematosus in the 24-year-old patient and rheumatoid arthritis in the 54-year-old patient. Both conditions were nearly quiescent when the patients manifested haemorrhagic diathesis. In response to combination therapy with prednisolone and cyclophosphamide, coagulation abnormalities were resolved together with complete elimination of factor VIII inhibitors in both patients. Thus, combination therapy with alkylating agents may be recommended as initial therapy for the management of autoimmune patients with factor VIII inhibitors. PMID- 11725832 TI - Fatal hydrothorax due to misplacement of a nasoenteric feeding tube. AB - Blind nasoenteric intubation was attempted in a patient with chronic parkinsonism. The tube was inadvertently misplaced and penetrated the left pleural cavity. The next day, the patient developed cardiopulmonary arrest during dietary supplement infusion. This complication ultimately led to the patient's death. We have reviewed the known complications of nasoenteric tube placement and conclude that difficult insertion in patients at risk from tube misplacement should be followed by chest radiography to confirm the correct placement of the tube before nutritional support is started. PMID- 11725833 TI - What kind of hepatitis? AB - Finding one major hepatotropic virus may not be enough to identify the aetiology of liver disease when risk factors are present, particularly in patients with past or present infection with other viral agents, or chronic liver disease. The pathogenic process in these cases is often complex. In the five cases we report, acute hepatitis (initiated by halothane, cytomegalovirus or Epstein-Barr virus) preceded the reactivation of hepatitis B infection, and these events occurred in patients with chronic hepatitis C infection. Each case demonstrates how several viruses can be implicated in the development of hepatitis, either as single agents or via cross-activation of T cells. The nosography of hepatitis, therefore, and the optimum therapeutic choices, can puzzle the clinical team. PMID- 11725834 TI - Recurrent pyogenic vertebral osteomyelitis associated with type 2 diabetes mellitus. AB - We report a case of recurrent pyogenic vertebral osteomyelitis associated with type 2 diabetes mellitus. A 51-year-old male was admitted to our hospital because of lumbago and general fatigue, with multiple ulcers on the soles of his feet. Staphylococcus aureus was isolated from peripheral blood and the foot ulcers, and 67Gallium scintigram showed abnormal isotope uptake, accumulated at the lower thoracic spine. Antibiotics were administered and the patient underwent intensive insulin therapy. Magnetic resonance imaging (MRI), performed after the levels of C-reactive protein decreased to 0.0 mg/dl, indicated old inflammatory changes at the Th8-Th9 spine and antibiotics were stopped. Unexpectedly, 8 days later the patient complained of lumbago with fever again, and MRI showed acute inflammatory changes at the same lesion site. This case report suggests that it is important for complementary antibiotic therapy to continue after signs of inflammation have disappeared in cases of pyogenic vertebral osteomyelitis. PMID- 11725835 TI - Reappraisal of the historic 1959 cat experiment in Minamata by the Chisso Factory. AB - Autopsy specimens from the historic cat experiment were recently discovered in a storage area at the Kumamoto University School of Medicine. The specimens were from an experiment prompted by physicians in the Chisso Minamata Plant following the announcement made by the Study Group for Minamata disease. On July 14, 1959 the Group announced that the disease was most likely caused by a kind of organic mercury. In order to prove or disprove that industrial waste from the Chisso Factory was the culprit in Minamata disease, a total of ten cats were fed food mixed with industrial waste produced in the acetaldehyde-producing plant. One of the ten cats, No. 717, was subsequently autopsied but the autopsy findings have never been published or recorded in the literature despite their historic significance. The rediscovered specimens were studied pathologically and biochemically, and were analyzed chemically with currently available techniques. Characteristic lesions of methylmercury poisoning were observed in the central nervous system, and the mercury levels in the cerebrum, cerebellum, liver and kidney were found to be markedly elevated in this animal. PMID- 11725836 TI - Enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR); its ability to differentiate Streptococcus pyogenes strains and applicability to the study of outbreaks of streptococcal infection. AB - We evaluated the ability of enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) to differentiate 95 Streptococcus pyogenes strains with M or T serotypes isolated from sporadic streptococcal infections as compared with M or T serotypings and pulsed-field gel electrophoresis (PFGE). Although the ERIC-PCR had less discriminatory power, defined as the ability to divide the strains with the same serotypes into the different sub-types, than PFGE, it consistently classified the strains into 16 patterns with a high correlation with M or T serotyping. The PCR method further discriminated 4 M or T serotypes into sub-types. The application of ERIC-PCR to 5 outbreaks of streptococcal infection produced the results that agreed closely with those of T serotyping and PFGE. ERIC-PCR has sufficient discriminatory power and is a quick and relatively easy technique, making it useful for routine epidemiological investigations. PMID- 11725837 TI - Restoring effect of human parathyroid hormone (1-34) on trabecular connectivity in ovariectomized rats. AB - The purpose of this study was to determine if the loss of the trabecular connectivities can be recovered by human parathyroid hormone-(1-34) therapy in ovariectomized (OVX) rats. Seven-month old female Wistar rats underwent ovariectomy or sham-operation at the beginning of the experiment. All sham operated groups (sham groups) were sacrificed after 0 (initial-BL group), 4, 8, 12, and 16 weeks, and one-third of the OVX rats were sacrificed as the baseline controls (OVX-BL groups) at 4, 8, and 12 weeks after OVX. Four weeks PTH or its vehicle treatment for residual OVX rats was started at 4, 8, or 12 weeks after OVX (OVX+PTH groups, OVX+vehicle groups). h-PTH (6.0 microg/kg) was injected subcutaneously six times a week for 4 weeks for each group. Their proximal right tibiae were processed for undecalcified Villanueva bone staining sections for bone histomorphometry. Furthermore, changes in trabecular connectivities were determined by node-strut analysis. h-PTH completely restored OVX-induced cancellous bone loss by stimulating bone formation. In node-strut analysis, node number in the OVX-BL-4, -8, and -12 groups was decreased to 53%, 49% and 44% of the initial-BL value respectively, and that in the OVX-4, -8, -12 + PTH groups recovered to 80%, 66%, 56% of the initial-BL value respectively. However, they were lower than those in their corresponding sham groups. The findings of this study suggested that decreased trabecular connectivity by OVX was recovered by intermittent h-PTH administration. However, delayed treatment blunted the restoration of trabecular connectivity. PMID- 11725838 TI - Glycoside in schizophrenic patients sera affects behavior of mouse forced swimming. AB - We found a glycoside in sera of schizophrenic patients. This glycoside increased climbing of mouse forced swimming and the climbing was decreased by dopamine D1 receptor antagonist SCH-23390 with a dose dependent manner. This glycoside had much reactivity of GalNAc alpha1-3GalNAc, but not that of polymannoses nor that of Gal beta1-3GlcNAc alpha1-serine or threonine. This strongly suggests that the glycoside is an isolated O-glycoside, but not N-glycoside nor O-glycoside having core 1 or 2. The present findings suggest that schizophrenic patients have the special glycoside affecting the D1 receptor activity in their serum. PMID- 11725839 TI - Estimation of nutrient intake by the new version of Japanese food composition tables in comparison with that by the previous version. AB - This study was initiated to examine if the revision of the food composition tables from the 4th version to the 5th version has induced any substantial changes in nutrient intake estimation, and if so, in which nutrient item and to what extent. For this purpose, 24-hour duplicates of food samples were collected in 1996 to 1998 from 71 girl university students, together with food menus of the day. The nutrients in the duplicates were estimated by use of the 4th and the 5th version food composition tables in parallel, with supplements of other databases. The results of the estimation with the two databases were compared by Student's paired t-test. Compared with the results with the 4th version, the estimation utilizing the 5th version gave 3 to 8% increases in intakes of major nutrients including energy (except for protein), a 20% increase in vitamin A, and decreases in iron (-13%) and NaCl (-3%), whereas there were only small or essentially no changes in other minerals and vitamins. The largest increment in energy through the re-calculation came from the cereals (especially rice), and it was meats for the increment in lipid intake. The increment in vitamin A with the 5th version is primarily due to the re-evaluation of fruits and vegetables. The present observation suggests that care should be practiced in examining secular trends in nutrition when the basis of estimation is shifted from one database of the 4th version of food composition tables to another of the 5th version. Further study of a larger scale is apparently warranted to confirm the conclusions. PMID- 11725840 TI - Comparison of the likelihood ratio and identity-by-state scoring methods for analyzing sib-pair test cases: a study using computer simulation. AB - To assess the power and significance of the likelihood ratio (LR) and the identity-by-state scoring (IBS) methods for a pair of siblings, we performed computer simulations by use of 10 DNA markers (HLA-DQalpha, D1S80, and 8 short tandem repeat loci) that were frequently analyzed in paternity tests in Japan. The combined power of discrimination of these 10 markers in the Japanese population is 0.999 999 999 98. Pedigrees each consisting of 10,000 pair of full siblings, half-siblings and unrelated individuals were generated and typed on all markers as random samples. Both the summation of log10 LR and IBS of each group had approximate standard normal distribution with significant differences between the means. Statistical studies showed that the LR method has 91% power to detect unrelated individuals and 38% power to detect half-siblings as not full-siblings with a 5% false-positive rate, whereas the IBS method does 87% and 42% powers, respectively. In 62% of full-siblings, in contrast with only 0.2% of unrelated individuals, the values of LR exceeded 100 which was equivalent to 0.99 of probability of full-sibship at 50% prior probability. The advantage of the LR method over IBS method was convincing especially for the detection of unrelated individuals as not half-siblings, however, the latter would be also informative for sib-pair tests if sufficient number of polymorphic markers are available. PMID- 11725842 TI - One more prescription. PMID- 11725841 TI - Chondroblastoma of a metacarpal bone mimicking an aneurysmal bone cyst: a case report and a review of the literature. AB - Chondroblastoma of the metacarpal bone has been extremely rare and only seven cases have been reported in the English literature. Here we reported the eighth case of a chondroblastoma that developed on the first metacarpal bone of the right hand of a 21-year-old man. Radiographs showed an expansile osteolytic lesion with a multilocular appearance. In MR images, the lesion showed low intensity in T1 and high intensity in T2-weighted images with multiple fluid fluid levels, which are findings resembling those of an aneurysmal bone cyst. From the pathological findings, however, it was recognized as a chondroblastoma with aneurysmal bone cyst-like change. Good clinical results was obtained by the subtotal resection of the metacarpal bone with a columnar-shaped iliac bone graft. PMID- 11725843 TI - Vertebroplasty--breakthrough in treatment of back pain. PMID- 11725844 TI - Drs. Cheryl and Roger Melkonian discuss the merits of the AACP application for specialty. American Academy of Craniofacial Pain. PMID- 11725845 TI - Dr. Abdel-Fattah criticizes the impairment rating in the AMA guides to the evaluation of permanent impairment. PMID- 11725846 TI - Vertical dimension. Part 1: comparison of clinical freeway space. AB - This study was conducted in order to compare the clinical freeway space measurements using three simple methods commonly used by dentists in their practices. The study was performed in 15 young healthy subjects with natural dentition and bilateral molar support. Artificial landmarks (adhesive tape) were placed on the more prominent parts of the nose and chin of each subject. Vertical dimension of occlusion (VDO) was measured in the intercuspal position. Postural vertical dimension (PVD) was measured in the following functional conditions: after swallowing saliva, after pronouncing the word "Mississippi", and in a relaxed postural mandibular position (RPMP). Then, the clinical freeway space value in each functional condition was obtained by subtracting VDO from PVD value. Significant differences among clinical freeway space values using three different methods were observed (ANOVA). A significantly higher clinical freeway space value was found using phonetics method than after swallowing and with the mandible in a relaxed postural position (Bonferroni multiple comparison test). No significant differences between swallowing and relaxed methods were found. These results seem to suggest that the measures of clinical freeway space depend upon the method used. PMID- 11725847 TI - Axial condyle morphology and horizontal condylar angle in patients with internal derangement compared to asymptomatic volunteers. AB - The purpose of this study was to evaluate the possible structural variations of axial condyle morphology and horizontal condylar angle between patients with internal derangement and asymptomatic volunteers. Forty-five symptomatic patients and 31 asymptomatic volunteers (AV) were included in this study. All subjects had bilateral high-resolution magnetic resonance imaging scans. Axial condyle morphology was evaluated using these images and classified into four diagnostic groups: convex, flat, and concave anterior surfaces, and tapered form. The angulation between the condylar long axis and the transversal plane was then measured. A Wilcoxon-Whitney-Mann-U-test demonstrated no significant differences in the horizontal angulation of the patients with disk displacement with reduction (DDR) (24.35+/-9.31) as compared to patients with displacement without reduction (DDN) (25.61+/-10.11) and asymptomatic volunteers (25.68+/-9.31) (AV). Eighty percent of the flat type condyles and 73.3% of the tapered type condyles belonged to the symptomatic patients with disk displacement with reduction (DDR) and without reduction (DDN). Chi-square test demonstrated significant differences between the diagnostic groups of condyle types (p<0.01). A comparison of the angular difference of right and left joints of asymptomatic volunteers and patients using the Wilcoxon-Whitney-Mann-U test showed that the results of the asymptomatic volunteers were significantly lower (p = 0.05). The conclusion that an increase in the horizontal condylar angle is a predisposing factor for the development of internal derangement or that the derangement develops during the prognosis of the disorder could not be clearly stated based on the results of this study. It is proposed that the difference between condyle morphology in patients with disk displacement and asymptomatic volunteers may be a result of the remodeling or degeneration caused by the disorder. PMID- 11725848 TI - Association of symptoms of TMD and orofacial pain with alexithymia: an epidemiological study of the Northern Finland 1966 Birth Cohort. AB - Alexithymia is a term denoting a deficit in the ability to differentiate emotional from physical states and to identify and describe one's feelings, as well as a preference for external oriented thinking. Alexithymia has been linked with various somatic and psychosomatic diseases, especially with chronic pain. The aim of this study was to evaluate the association between alexithymia and symptoms of temporomandibular disorders (TMD) as well as oro-lingual and dental pain, in a large representative population sample of young adults. The study was a part of the 31-year follow-up study of the Northern Finland Birth Cohort originally consisting of 12058 live births in the year 1966. In 1997, 4893 subjects living in northern Finland or in the capital area, who participated in a field study of the project and later returned a postal questionnaire, made up the sample of this study. Information concerning symptoms of TMD and oro-lingual and dental pain was collected from the subjects. To assess alexithymia, the Toronto Alexithymia Scale-20 (TAS-20) was used. In addition, information about depression, marital status and self-rated health was collected. The proportion of alexithymics (TAS score over 60) was higher in subjects with the most orofacial symptoms than in asymptomatic subjects. In men, alexithymia associated significantly with facial pain, difficulties in mouth opening, oro-lingual pain and dental pain, and in women with pain on jaw movement and dental pain. After adjusting for depression, marital status, and self-rated health, a significant association remained between alexithymia and the symptoms mentioned, except for facial pain in men. It can be concluded that alexithymia is connected with orofacial symptoms. Clinicians treating these symptoms should be familiar with the concept of alexithymia. PMID- 11725849 TI - Variable site of oropharyngeal narrowing and regional variations of oropharyngeal collapsibility among snoring patients during wakefulness and sleep. AB - The oropharynx shows rhythmic caliber changes in accordance with the respiratory cycle. Dynamic imaging of the oropharynx is required to evaluate airway changes associated with snoring and obstructive sleep apnea. The purpose of this study was to determine the sites of narrowing/obstruction and to measure the regional collapsibility of the oropharynx during respiration during the sleep and awake cycles in snoring patients. Seven snoring patients were included for this study. Electron Beam Tomograms of the oropharynx were taken while the patient was awake and asleep, and dynamic views were reconstructed. The site of narrowing or obstruction varied widely among patients and differed from awake to asleep. The mean of smallest minimum cross-sectional area of the oropharynx was 60.38+/-42.97 mm2 in the awake state and 16.90+/-20.43 mm2 in the asleep state. Although the collapsibility of the oropharynx showed considerable regional variations, it increased during sleep at the overall oropharynx. The low retropalatal region showed the smallest minimum cross-sectional area and the highest collapsibility; however, the most prominent changes in the minimum cross-sectional area and collapsibility during sleep were observed in the high retroglossal region. PMID- 11725850 TI - MRI findings concerning the lateral pterygoid muscle in patients with symptomatic TMJ hypermobility. AB - Clinical studies have shown a close association between temporomandibular joint hypermobility (TMJH) and temporomandibular disorders (TMD). While pathological change of the lateral pterygoid muscle (LPM) is one of the most emphasized in studies of TMD, there have been no detailed clinical reports of the LPM studies using magnetic resonance imaging (MRI) in TMJH. This study investigates structural and pathological alterations involving the LPM in patients with TMJH using MRI. A retrospective analysis was made of high-field MRI images from 98 patients with TMJH. LPMs of 143 joints were analyzed. In 110 joints (77%), hypertrophy, atrophy, and contracture were found in the superior belly and/or the inferior belly of the LPM. Pathological changes were more frequently found in the superior rather than the inferior belly of the LPM. In the cases with abnormalities in both bellies of the LPM, hypertrophy of the inferior belly was usually found combined with other changes of the SBLPM. The results of this study indicated that the pathological changes of the LPM or MRI are not infrequent in patients with symptomatic TMJH. PMID- 11725851 TI - Migraine and tension-type headache reduction through pericranial muscular suppression: a preliminary report. AB - Migraine and tension-type headaches have always plagued mankind. In spite of all the research dollars spent trying to determine the etiologies of these headaches, the neurology community still has not established a known cause of migraine and tension type headaches. This paper describes a study that was conducted for the U.S. Food and Drug Administration in which the efficacy of the Nociceptive Trigeminal Inhibition Tension Suppression System was evaluated and proved safe and efficacious in the reduction of medically diagnosed migraine and tension-type headache. PMID- 11725853 TI - Posterior disk displacement of the TMJ: MRI evidence in two cases. AB - Posterior disk displacement is a rare temporomandibular joint (TMJ) disorder. The main clinical sign is sudden molar open-bite (jaw locked in the open position). This may be accompanied by a sensation of intra-articular foreign body and more rarely joint pain. Joint sounds are unremarkable. Mouth opening may be slightly impaired. Hypothetically, like anterior disk displacements, posterior disk displacements can be classified as either reducible or nonreducible. A definitive diagnosis requires magnetic resonance imaging (MRI). There is no consensus concerning treatment. A conservative course of treatment can be successful in patients with functional impairment and should always be attempted before irreversible, invasive therapy. PMID- 11725852 TI - The effects of a home exercise program on pain and perceived dysfunction in a woman with TMD: a case study. AB - There are few reports analyzing the effects of exercise on patients with temporomandibular disorders. This paper presents a case study examining whether there were additional benefits from performing neuromuscular control home exercises (NMCHE) in a patient with temporomandibular disorder who was already receiving conventional treatment. A woman, 41 years of age, completed a health status questionnaire called the TMJ Scale prior to being accepted. She completed additional TMJ Scales after receiving conventional treatment and again after conventional treatment was combined with neuromuscular control home exercises. Based on the TMJ Scale's best subscore indicator of the presence or absence of a temporomandibular disorder, the patient did not derive benefit from conventional treatment without exercise. However, she received a large benefit from the addition of NMCHE. It was concluded that exercises for patients with temporomandibular disorders may be beneficial to those who do not improve with conventional treatment alone. PMID- 11725854 TI - Analyses of muscular activity, energy metabolism, and muscle fiber type composition in a patient with bilateral masseteric hypertrophy. AB - Hyperwork of the masseter muscles due to habitual parafunction is thought to induce masseteric hypertrophy (so called work hypertrophy). However, the causes underlying this disease are not yet fully understood. Recently, we had a patient with bilateral masseteric hypertrophy, and we performed a partial excision of the masseter muscles. In this patient's case, we examined muscular activity, energy metabolism, and fiber type composition of the masseter muscles using electromyograms (EMG), 31P-magnetic resonance spectroscopy (MRS), and enzyme histochemistry. The EMG showed no hyperactivity, and the 31P-MRS showed normal energy spectral patterns and PCr contents of the masseter muscles. The fiber type composition, however, in the muscles in this case was very different from that in muscles with "work hypertrophy" and also that in normal masseter muscles: 1. Loss of type IIB fibers; 2. Increases in type IIA and in type IM & IIC fibers; and 3. Decrease in type I fibers. The findings suggest that this is not a case of work hypertrophy but a case of compensatory hypertrophy possibly due to a lack of high tetanus-tension type IIB fibers. PMID- 11725855 TI - Craniofacial pain as a dental specialty: a white paper by the American Academy of Craniofacial Pain. PMID- 11725856 TI - Neuroimaging in communication sciences and disorders: an introduction. PMID- 11725857 TI - Neuroimaging studies of speech an overview of techniques and methodological approaches. AB - Over the past two decades, there has been an explosion in the use of imaging technology to study the structure and function of the human brain. The purpose of this article is to explore how functional neuroimaging has been applied to the study of speech production. This article begins with a brief review of neuroimaging methods and limitations. Then, two approaches that can be used to study the brain areas that support speech production are illustrated. The first approach is based upon comparisons across different types of language production tasks; the second approach is based upon comparing the effects of different types of stimuli within a single task. Results obtained using these approaches will be used to dissociate the contributions of different brain regions involved in speech production. For example, evidence will be presented that Broca's area contributes to phonological encoding, whereas motor cortex, the supplementary motor area (SMA), and the cerebellum support phonetic encoding and articulation. LEARNING OUTCOMES: As a result of this activity, the participant will be able to describe basic methods for conducting a positron emission tomography study and a functional magnetic resonance imaging study. The participant will also be introduced to two approaches for fractionating the brain regions involved in speech production: (1) comparisons between tasks, and (2) manipulations of stimulus materials. Finally, the participant will be able to summarize a cognitive model of the components of speech production that will be introduced, and potential mappings between these components and particular brain regions will be discussed. PMID- 11725858 TI - The neural representation of language in users of American Sign Language. AB - Studies of American Sign Language (ASL) offer unique insights into the fundamental properties of human language. Neurolinguistic studies explore the effects of left and right hemisphere lesions on the production and comprehension of signed language. Following damage to the left hemisphere perisylvian regions, signers, like users of spoken languages, exhibit frank aphasic disturbances. Sign language paraphasia illustrates the linguistic specificity of impairment. A case study involving cortical stimulation mapping (CSM) in a deaf signer provides evidence for the specialization of Broca's area in sign language production. The effects of right hemisphere damage highlight the specialized properties of sign language use. Data from functional magnetic resonance imaging (fMRI) of deaf signers confirm the importance of left hemisphere language structures in the use of signed language, but also reveal the contributions of right hemisphere regions to the processing of ASL. These studies provide new insights into the complementary roles of biology and environment in language representation in the human brain. LEARNING OUTCOMES: As a result of this activity, the participant will read studies of aphasia in users of signed language and a discussion of neurolinguistic studies of paraphasia in ASL. The participant will examine the role of the right hemisphere in language use and findings from a functional imaging study of sentence processing in ASL and English. PMID- 11725859 TI - Positron emission tomography in cochlear implant and auditory brainstem implant recipients. AB - Positron emission tomography (PET) imaging was used to evaluate the brain's response to auditory stimulation, including speech. Five cases of adults illustrate the utility of PET to illuminate auditory processing with cochlear or brainstem implants. Subjects showed varying degrees of success in processing speech, which was reflected in the resulting PET images. Functional speech processing was associated with activation in areas classically associated with speech processing. In one patient who did not achieve functional speech processing, activation in frontal regions suggests that the subject used other cognitive strategies to assist auditory processing. LEARNING OUTCOMES: As a result of this activity, the participant will be introduced to the principles of PET and the application of this technology to deaf patients who have received an implanted auditory prosthesis. PMID- 11725860 TI - Neurobiological studies of reading and reading disability. AB - Evidence from neuroimaging studies, including our own, suggest that skilled word identification in reading is related to the functional integrity of two consolidated left hemisphere (LH) posterior systems: a dorsal (temporo-parietal) circuit and a ventral (occipito-temporal) circuit. This posterior system appears to be functionally disrupted in developmental dyslexia. Relative to nonimpaired readers, reading-disabled individuals demonstrate heightened reliance on both inferior frontal and right hemisphere posterior regions, presumably in compensation for the LH posterior difficulties. We propose a neurobiological account suggesting that for normally developing readers, the dorsal circuit predominates at first, and in conjunction with premotor systems, is associated with analytic processing necessary for learning to integrate orthographic with phonological and lexical semantic features of printed words. The ventral circuit constitutes a fast, late-developing, word form system, which underlies fluency in word recognition. LEARNING OUTCOMES: As a result of this activity, (1) the participant will learn about a model of lexical processing involving specific cortical regions. (2) The participant will learn about evidence which supports the theory that two dorsal LH systems may be disrupted in developmental dyslexia. (3) The participant will learn that individuals with reading impairment may rely on other regions of the brain to compensate for the disruption of posterior function. PMID- 11725861 TI - Brain imaging studies of developmental stuttering. AB - This paper reviews recent brain imaging research on stuttering against a background of studies that the writer and colleagues have been conducting at the University of Texas Health Science Center in San Antonio. The paper begins by reviewing some pertinent background to recent neuroimaging investigations of developmental stuttering. It then outlines the findings from four brain imaging studies that the San Antonio group has conducted using H2(15)O positron emission tomography (PET). Finally, some of the principal findings that are emerging across brain imaging studies of stuttering are reviewed, while also highlighting- and attempting to resolve--some apparent across-study inconsistencies among the findings. Research on stuttering using magnetoencephalogaphy (MEG) and transcranial magnetic stimulation (TMS) is also considered. The findings increasingly point to a failure of normal temporal lobe activation during speech that may either contribute to (or is the result of) a breakdown in the sequencing of processing among premotor regions implicated in phonologic planning. LEARNING OUTCOMES: As a result of this activity, the participant will become familiar with some recent neurophysiological correlates of stuttering and what they suggest about the nature of this disorder. PMID- 11725862 TI - Role of the sarcoplasmic reticulum Ca2+-ATPase on heat production and thermogenesis. AB - The sarcoplasmic reticulum of skeletal muscle retains a membrane bound Ca2+ ATPase which is able to interconvert different forms of energy. A part of the chemical energy released during ATP hydrolysis is converted into heat and in the bibliography it is assumed that the amount of heat produced during the hydrolysis of an ATP molecule is always the same, as if the energy released during ATP cleavage were divided in two non-interchangeable parts: one would be converted into heat, and the other used for Ca2+ transport. Data obtained in our laboratory during the past three years indicate that the amount of heat released during the hydrolysis of ATP may vary between 7 and 32 kcal/mol depending on whether or not a transmembrane Ca2+ gradient is formed across the sarcoplasmic reticulum membrane. Drugs such as heparin and dimethyl sulfoxide are able to modify the fraction of the chemical energy released during ATP hydrolysis which is used for Ca2+ transport and the fraction which is dissipated in the surrounding medium as heat. PMID- 11725863 TI - Mechanism of thyroid-hormone regulated expression of the SERCA genes in skeletal muscle: implications for thermogenesis. AB - Thyroid hormone increases the Ca2+-ATPase activity of the sarcoplasmic reticulum (SR) in skeletal muscle, thereby increasing the energy-turnover associated with Ca2+-cycling during contraction and rest. The fast-muscle isoform of the Ca2+ ATPase (SERCA1) and the slow-muscle isoform (SERCA2a), are encoded by two genes that are transcriptionally regulated by T3. The SERCA1 isoform can be expressed to considerably higher levels than the SERCA2a isoform. The stimulation of transcription of the SERCA1 gene by T3 is mediated by two thyroid hormone response elements, located in the promoter of this gene. The intracellular [Ca2+] can modulate the effect of T3. The increase in SR Ca2+-ATPase activity seen when T3-levels rise above normal, results from the induction of SERCA1 expression in slow muscle fibers. Concomitant high levels of Ca2+-ATPase activity are associated with down-regulation of SERCA2a expression in these fibers. The observed T3-dependent increase in SERCAI expression and associated Ca2+ATPase activity will increase the overall metabolic rate of the organism significantly under normal conditions, because of the high average level of contractile activity of slow fibers. Given the rise in serum T3-levels during prolonged cold exposure, these data suggest that fiber-specific stimulation of SERCA1 expression contributes to the thermogenic response in non-shivering thermogenesis. This mechanism may be particularly relevant in larger mammals, which have a relatively high percentage of slow fibers in skeletal muscle, and which need to rely on tissues other than brown fat for the generation of extra heat. PMID- 11725864 TI - Malignant hyperthermia: an inherited disorder of skeletal muscle Ca+ regulation. AB - Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle characterized by muscle contracture and life-threatening hypermetabolic crisis following exposure to halogenated anesthetics and depolarizing muscle relaxants during surgery. Susceptibility to MH results from mutations in Ca2+ channel proteins that mediate excitation-contraction (EC) coupling, with the ryanodine receptor Ca2+ release channel (RyRI) representing the major locus. Here we review recent studies characterizing the effects of MH mutations on the sensitivity of the RyRI to drugs and endogenous channel effectors including Ca2+ and calmodulin. In addition, we present a working model that incorporates these effects of MH mutations on the isolated RyRI with their effects on the physiologic mechanism that activates Ca2+ release during EC coupling in intact muscle. PMID- 11725865 TI - Heat production during anesthetic-induced malignant hyperthermia. AB - Malignant hyperthermia (MH) is a pharmacogenetic disease which predisposes to the trigger of a life-threatening, hypermetabolic syndrome by potent inhaled anesthetics and by depolarizing skeletal muscle relaxants. Heat production in the anesthetized MH can be profound with 5-fold increases in oxygen consumption. The trigger anesthetics cause an abnormal, sustained rise in myoplasmic calcium levels. Possible mechanisms by which continuous release of calcium from skeletal muscle sarcoplasmic reticulum stores can produce the profound hyperthermia are discussed. Mutations in the gene coding the ryanodine receptor calcium release channel have been found in MH families and these mutant channels may be the functional basis for MH. PMID- 11725866 TI - Thermogenesis in birds. AB - The article discusses the importance of avian skeletal muscle as a source for heat generation by means of both shivering and non-shivering. Non-shivering thermogenesis in birds is still a polemic issue. Recent evidence at the molecular/cellular level indicates, however, that this type of heat generation may also exist among birds. The involvement of the sarcoplasmic reticulum calcium ATPase in non-shivering thermogenesis is discussed in-depth. PMID- 11725867 TI - Barbara Cannon's data on the UCP1-ablated mice: "non-cannonical" point of view. AB - The data of Cannon and co-workers on UCP1-ablated mice are interpreted assuming that UCP2 and UCP3 are involved in thermoregulation as fatty acid-dependent uncouplers although they are not sufficient, in the absence of UCP1, for long term maintenance of normal body temperature of mice after sudden and strong decrease in the ambient temperature. I would like to suggest that in brown fat of control mice, UCP1 is present in an amount higher than UCP2 and 3 and, therefore, is able to cause (a) some fatty acid-mediated decrease in proton motive force in resting state and, hence, (b) oxidation of CoQH2 to CoQ which is shown by Klingenberg and coworkers to be cofactor for UCPs. This results in strong uncoupling and thermogenesis mediated by UCP1, 2 and 3. In the UCP1-ablated mice, activity of UCP2 and 3 appears to be insufficient to induce CoQH2 oxidation in resting brown fat mitochondria, which results in hypothermia. PMID- 11725868 TI - The discovery of an uncoupling mitochondrial protein in plants. AB - This paper describes peculiar properties of plant mitochondria and summarizes the experiments that led to the discovery of an uncoupling protein in these mitochondria. Recent advances in the study of the biochemical and physiological properties as well as on genes encoding plant uncoupling proteins are described in articles by Borecky et al., Jezek et al., and Jarmuszkiewicz et al. in this issue. PMID- 11725869 TI - Mitochondrial uncoupling proteins in mammals and plants. AB - Uncoupling proteins (UCPs) belong to a distinct cluster of the mitochondrial anion carrier family. Up to five different uncoupling protein types were found in mitochondria of mammals and plants, and recently in fishes, fungi and protozoa. They exhibit a significantly conserved structure with several motifs specific to either the whole cluster or protein type. Uncoupling proteins, as well as the whole mitochondrial anion carrier gene family, probably emerged in evolution before the separation of animal, fungi, and plant kingdoms and originate from an anion/nucleotide or anion/anion transporter ancestor. Mammalian UCP1, UCP2, UCP3, and plant uncoupling proteins pUCP1 and pUCP2 are similar and seem to form one subgroup, whereas UCP4 and BMCP1 belong to a different group. Molecular, biochemical, and phylogenic data suggest that UCP2 could be considered as an UCP prototype. UCP1 plays its biological role mainly in the non-shivering thermogenesis while the role of the other types is unknown. However, hypotheses have suggested that they are involved in the general balance of basic energy expenditure, protection from reactive oxygen species, and, in plants, in fruit ripening and seed ontogeny. PMID- 11725870 TI - Alternative oxidase and uncoupling protein: thermogenesis versus cell energy balance. AB - The physiological role of an alternative oxidase and an uncoupling protein in plant and protists is discussed in terms of thermogenesis and energy metabolism balance in the cell. It is concluded that thermogenesis is restricted not only by a lower-limit size but also by a kinetically-limited stimulation of the mitochondrial respiratory chain. PMID- 11725871 TI - Biophysics and physiology of temperature regulation in thermogenic flowers. AB - The flowers or inflorescences of certain primitive seed plants are able to regulate their temperature during blooming by modulating the rate of heat production to remain much warmer than the surroundings. A large drop in ambient temperature causes a smaller drop in flower temperature which causes an increase in the rate of heat production by futile involvement of the cytochrome and alternative oxidase respiratory pathways. The result is that the rate of heat production is inversely related to ambient temperature and flower temperature remains high and relatively independent of ambient temperature. While the biophysics of thermal balance in the whole flowers is better understood, the regulation of the biochemical heat-generating pathways is not known. PMID- 11725872 TI - Possible basic and specific functions of plant uncoupling proteins (pUCP). AB - Evidence has been provided that the plant uncoupling proteins (pUCP) play basic physiological roles similar to the other uncoupling protein subfamily members (mammalian UCP1,2,3,4 and BMCP) and are effective in the situations of slight uncoupling that leads to: (1) accelerated respiration and metabolic rates that are beneficial to plant growth and development; (2) decreased formation of reactive oxygen species in mitochondria; and, (3) mild thermogenesis, inevitably accompanying the previous two phenomena. Hypothetically, specific physiological roles of pUCP such as cut off of ATP synthesis could be manifested in connection with climacteric respiratory rise during fruit ripening, seed dormancy, and plant senescence. pUCP might also facilitate growth under low temperatures, e.g., during seed germination or in roots. The existence of these specific roles is suggested by the immunochemical and functional localization of pUCP in mitochondria of fruits, seeds and roots of various plant species. PMID- 11725873 TI - 2001 Founders Award. J. Newell Stannard. PMID- 11725874 TI - 2001 Founders Award. Paul L. Ziemer. PMID- 11725875 TI - 2001 Elda E. Anderson Award. Tsz-Yik Edmond Hui. PMID- 11725876 TI - 2001 Distinguished Scientific Achievement Award. John H. Hubbell. PMID- 11725877 TI - 2001 Outstanding Science Teacher Award. Michael A. Mocherman. PMID- 11725878 TI - 2001 Robley D. Evans Commemorative Medal. Allen Brodsky. PMID- 11725879 TI - 2001 William B. McAdams Outstanding Service Award. George J. Vargo, Jr. PMID- 11725880 TI - 2001 National Registry of Radiation Protection Technologists Arthur F. Humm, Jr., Memorial Award. Paul W. Harvey. PMID- 11725881 TI - The whistleblower implications of radiation injury lawsuits: lessons learned from In re McCafferty. AB - While it is widely known that strict regulations protect nuclear workers who raise safety-related concerns, few are aware that the riling of a claim for radiation-related injuries has been interpreted to constitute a "protected activity" under the Energy Reorganization Act, which endows the claimant with "whistleblower" status. This means that negative employment-related actions taken against a radiation injury claimant can result in detrimental, even draconian, consequences for the employer involved. The case of In re McCafferty illustrates this vividly. The claimants in McCafferty were six contract insulators who filed lawsuits for emotional distress they claimed to have suffered due to exposures they received at Centerior Energy's Davis-Besse nuclear plant. Because of the pending lawsuit and claim for emotional distress, Centerior denied all six claimants access to Centerior's plants, which resulted in their being terminated by their employer. The claimants brought an action with the Department of Labor, succeeded in convincing the Administrative Review Board that Centerior's actions constituted retaliation under the ERA "whistleblower" provision, and were afforded nearly full relief on their claims. This article explains how Centerior's actions ran afoul of the Energy Reorganization Act's "whistleblower" provision. It also describes what licensees and their subcontractors can do if faced with similar circumstances. PMID- 11725882 TI - Radiation litigation and the nuclear industry--the experience in the United Kingdom. AB - In the United Kingdom, the Nuclear Installations Act 1965 places a "strict" statutory duty on the operators of nuclear facilities to ensure that any exposure to radiation resulting from operations does not cause injury or damage. A claimant does not have to prove fault to receive compensation under the Act, only causation. The 1965 Act has been fundamental in shaping litigation involving the nuclear industry in the UK. Civil law cases brought under the Act will be heard before a single judge (with no jury or technical assessor) who must present his or her decision in a reasoned judgment. This process leads to a considerable volume of expert evidence being presented to the court and extensive cross examination of witnesses. The expense and uncertain outcome of cases involving claims by nuclear workers that occupational exposure to radiation had caused the development of cancer has led to employers and trade unions setting up the voluntary Compensation Scheme for Radiation-linked Diseases as an alternative to litigation. This Scheme has worked well and is held up as a model of alternative dispute resolution. However, a few cases concerning personal injury or damage to property have come before the courts when the defendant nuclear operator considered that the claims were technically unjustified and where settlement was not a policy option. As anticipated, these cases were lengthy, complex, and expensive. The radiation doses assessed to have been received by the individuals who were the subject of claims, whether workers or members of the public, have been crucial to the outcome. The technical expertise of health physicists and allied specialists has been vital in establishing defensible estimates of dose, and this contribution can be expected to remain of high importance in radiation litigation in the UK. PMID- 11725883 TI - Spread of 60Co contaminated steel and its legal consequences in Taiwan. AB - Since 1992, over 200 civilian residential and school buildings in Taiwan have been identified to have contained 60Co contaminated steel rebar emitting excessive gamma-radioactivity in living environments. These buildings were mostly constructed in early 1983 and 1984 by employing steels from one steel mill, which had recycled unknown 60Co orphan sources in northern Taiwan. In 1994, a group of residents who once stayed for a protracted period up to 10 y in the contaminated Ming-Sheng Villa filed a civil action against Taiwan's nuclear regulatory office, the Atomic Energy Council, for state tort compensation of 3.4 M U.S. dollars in equivalent. After three years of court processes, the Taipei District Court handed down a decision in partial favor of the exposed residents. Both parties soon appealed against this judgment to the Taiwan Appellate Court. This article analyzes the main legal issues involved, including government's obligations to prevent and eliminate contamination, to take preventive measures, and to take necessary remedial measures; and plaintiffs' assertion on any legal right against governmental offices. Moreover, discussion issues contain the scope of damage and compensation, causation analysis, absence of effective and efficient regulation over radioactive contamination, limit of tort compensation law and compensation amount, weight of medical evidence as well as role of expert witnesses, and related comparative legal studies. PMID- 11725884 TI - The role of the radiation safety specialist as witness: risk communication with attorneys, judges, and jurors. AB - As nuclear workers and members of the public continue to fear radiation in this litigious society, specialists in radiation safety will often be called upon as experts to explain the significance of radiation exposures or as fact witnesses to explain radiation safety practices. Radiation risk communication with attorneys, judges, and jurors presents special challenges to the communication skills of health physicists. Your role as the radiation specialist is to present testimony, either in the form of a deposition or as a trial witness, in a way that a judge or jury can understand. As a specialist in radiation safety, you will also need to educate the attorney that you work with so that he or she can ask the right questions and defend challenges in the case. The way that you communicate to attorneys, judges, and jurors could have a great impact on the case's outcome. As a radiation specialist, your testimony is not only to present the scientific basis for radiation health risks, but also to persuade the judge or jurors in the direction of the desired outcome of the case. Insights from the Myers-Briggs Type Indicator show that judges and jurors are most likely persuaded by "Sensing" language that is specific, detailed, measurable, and verifiable with their five senses. Thus, the conceptual, abstract, and theoretical "Intuitive" language often favored by radiation experts may not be understood or appreciated by a judge or jurors. They may also prefer the more personal, empathetic, and caring "Feeling" language rather than the impersonal, logical, and analytical "Thinking" language favored by health physicists. People's feelings about radiation risks are a big factor in radiation cases and providing testimony to address feeling-based conclusions requires a very different communication approach than normally used by health physicists. An understanding of language preferences can be crucial for effective communication with attorneys, judges, and jurors. These insights are derived from the author's experience as a communication specialist and as a radiation expert for the plaintiffs in two radon cases. This paper also provides insights into the qualifications for serving as an expert or fact witness, preparation for a trial, presenting testimony, the courtroom as drama, and the best language modes for persuasive communications with judges and jurors. PMID- 11725885 TI - Application of the Supreme Court's Daubert criteria in radiation litigation. AB - In 1993, the U.S. Supreme Court set forth the standard for determining the admissibility of expert scientific evidence in litigation. This standard is known as the Daubert criteria, named after the pertinent case, Daubert v. Merrell Dow Pharmaceuticals, Inc. The Daubert criteria require the courts to determine whether an expert's testimony reflects scientific knowledge, whether his/her findings are derived by the scientific method, and whether the work product is based on good science. The Daubert criteria are especially important in radiation litigation because issues involving radiation doses and effects are often complex and thus a jury will typically rely heavily on the analysis and opinions of experts. According to the Daubert criteria, scientific opinions must be based on a methodology that has a valid, testable hypothesis; has been subject to peer review; and is generally accepted in the scientific community. Additionally, the expert must be qualified to present opinions based on the methodology. Although the application of the Daubert criteria in radiation litigation is highly dependent on the specific court and judge presiding over the case, there have been recent high-profile cases in which application of the criteria has resulted in the dismissal of analysis and opinions offered by scientific experts. Reasons for the dismissals have included basic scientific errors such as failure of the expert to consider all possible explanations for an observed phenomenon, the selective use of data by the expert, and the failure to acknowledge and resolve inconsistencies between the expert's results and those of other investigators. This paper reviews the Daubert criteria as they apply to radiation litigation and provides examples of the application of the criteria from recent judgments involving the Three Mile Island and Hanford Downwinders cases. PMID- 11725886 TI - Admissibility of scientific evidence post-Daubert. AB - Proof of medical causation is the key element in cases involving alleged radiation injury. Until 1993, the use of scientific testimony in the courtroom was governed by the Frye doctrine requiring that a theory be "generally accepted" before it can be the basis of an expert's opinion. Applying that rule trial courts adopted a "let it all in" approach resulting in a near overdose of pseudoscience presented to juries. With its decision in Daubert v. Merrell Dow Pharmaceuticals, Inc., 509 U.S. 579, 113 S. Ct. 2786 (1993), the U.S. Supreme Court announced a new five-factor, non-exclusive test for federal district courts to consider when assessing the opinion of scientific experts. The Court further directed that the trial courts, acting as a gatekeeper for expert evidence, must evaluate whether there is an adequate "fit" between the expert's data and the opinion offered. This article examines how the Daubert standard has been implemented in federal court to combat junk science. Examples from recent case law dealing with operational health physics issues are presented as an illustration of the use of the five-factor test to challenge questionable testimony on causation. Guidance is offered to prospective expert witnesses in radiation-related litigation to insure that proposed testimony will be admissible in district court. PMID- 11725887 TI - Pre-litigation strategies--gathering and preserving documentary evidence. AB - Radiation injury claims may arise under various legal theories. In addition, plaintiffs may advance such claims within different jurisdictional venues, such as federal and state courts and workers' compensation boards. Irrespective of the jurisdiction or the legal theory underlying the claim, one element remains common to these claims--the quality and quantity of the evidence. While many different pieces of evidence may be needed to litigate a radiation injury claim, the most important evidence for the investigating health physicist is that which establishes the nature and extent of radiation exposure. Most radiation injury claims are associated with late radiation injury, often an allegation of radiation-induced cancer. Because radiation-induced cancers have a long latency period, claims may not arise for years, or even decades, after exposure. Therefore, the immediate challenge to the health physicist, who investigates an exposure, is to avoid the temptation of a "wait and see" approach to gathering evidence. Not only may evidence be short-lived in nature, but with the passage of time memories grow dim and witnesses may become unavailable. Prompt and thorough gathering of pertinent evidence likely will be a determining factor in the outcome of any radiation injury claim. Although ensuring the availability of all pertinent evidence is the key role of the investigating health physicist, he or she also can help to ensure that the evidence does not inadvertently become inadmissible in a court of law, for example, under the hearsay rule. To ensure that the necessary evidence is available in admissible form, the task of gathering evidence should be systematically approached using a pre-established process that reflects a basic understanding of the rules of evidence. Such a process is discussed here. PMID- 11725888 TI - Review of 239Pu and 226Ra effects in beagles. AB - A long term biological study has been completed that was designed to assess the predicted effects in humans of internally deposited 239Pu by comparison with 226Ra in beagles. Herein we summarize for the first time results of several previous reports about the effects of these two radionuclides in our beagles in an attempt to elucidate what has been learned since the beginning of the study in the early 1950's. Perhaps the most important finding was that bone surface seeking plutonium is more toxic at equal mean skeletal radiation doses (<3 Gy for 239Pu, <20 Gy for 226Ra) than bone volume-seeking radium for the induction of skeletal malignancy by about a factor of 16 for a single intravenous injection of monomeric 239Pu. In addition, ancillary studies have shown that when plutonium transfers continuously onto bone surfaces from a depot of particulate 239Pu in phagocytic cells, its relative toxicity per Gy average skeletal dose is enhanced by about a factor of 2. Juvenile animals or dogs injected as mature adults were only about half as sensitive for equal mean skeletal doses as dogs injected as young adults. Male and female dogs were about equally sensitive to radiation of the skeleton by either radionuclide. Findings about radiation-induced fractures are summarized as well as data on the induction of soft-tissue malignancies by 239Pu or 226Ra. Natural survival was not affected at the lower dosage levels of either 226Ra or 239Pu as compared with control dogs given no radioactivity, but the survival of animals at higher levels was reduced. No additional life shortening effects beyond those attributable to occurrence of radiation-induced malignancies or other radiation-induced effects were suggested by analysis of data for low dosage levels. PMID- 11725889 TI - Radionuclides in Peconic River fish, mussels, and sediments. AB - For regulatory oversight and quality control of Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) actions, fish, mussels, and sediments were analyzed from the Peconic River system on Long Island, NY, downstream of the Brookhaven National Laboratory, as well as from control locations. The analyses were for photon-emitting radionuclides (notably 60Co and 137Cs), uranium, plutonium, and americium. Sediments were cored in 4 sections to 0.37 m depth, whole fish were analyzed, and mussels were separated into flesh and shells. Radioisotopes of the cited elements were detected in sediment, some of the fish contained 137Cs, 241Am, and uranium, and mussel flesh contained 137Cs and uranium. All of the 60Co, 233U, and enriched uranium, and some of the 137Cs and 241Am, can most likely be attributed to Brookhaven National Laboratory. The other radionuclides (and some of the 137Cs and 241Am) are believed to have either fallout or nature as their origin. The New York State Department of Health (NYSDOH) evaluated the radiological data in terms of adverse health implications due to consumption of fish with the levels of reported radioactivity. The NYSDOH determined that the added radiation doses likely to result from eating this fish are a small fraction of the radiation dose that normally results from radionuclides present in the body from natural sources. PMID- 11725890 TI - Non-invasive measurement of water content in lung phantom by neutrons: latest improvements to the technique. AB - The novel technique for measuring the water content in lungs based on the slowing down of fast neutrons has been improved making possible its clinical use on patients. This goal has been reached by performing computer simulations that closely mimic the experimental setup and by acquiring a new set of measurements obtained with a plutonium-beryllium (Pu-Be) neutron source provided of an activity suitable to clinical requirements. Finally, accurate radiation dose measurements have been carried out to ensure the acceptability of the technique. PMID- 11725891 TI - An evaluation of germanium detectors employed for the measurement of radionuclides deposited in lungs using an experimental and Monte Carlo approach. AB - A study was undertaken to evaluate the performance of an advanced design broad energy germanium detector for the in vivo measurement of radionuclides in lungs. Relative counting efficiency, background, and sensitivity for lung counting arrays consisting of four, three, and two 80-mm-diameter by 20-mm-thick (80 x 20 mm) broad energy germanium detectors were simulated by collecting spectra with the single 80 x 20 mm broad energy germanium at each of four locations over a humanoid torso phantom. Regions of interest were evaluated for photon energies ranging from 17 to 1,500 keV. The 80 x 20 mm detector arrays were then benchmarked against a standard array of four 70-mm-diameter by 20-mm-thick (70 x 20 mm) broad energy germanium detectors. Since testing new equipment can be an expensive and time consuming process, an alternative approach, using Monte Carlo simulations instead of physical measurements, was also evaluated and compared to experimental data. With this approach, counting efficiency and minimum detectable amount were simulated for two sizes of germanium detectors (70 mm and 80 mm diameter) at four different crystal thicknesses (15, 20, 25, and 30 mm). For the experimental measurements, arrays consisting of three and four 80 x 20 mm broad energy germanium detectors resulted in an increase in counting efficiencies, relative to the standard array, at all photon energies. The greatest relative increase was observed for the four-detector array (24-35%). In contrast, counting efficiency decreased, relative to the standard array, by 24-28% with a two detector array. Arrays consisting of two and three 80 x 20 mm broad energy germanium detectors resulted in decreased relative background at all photon energies, with the exception of the 946 keV photon for the three-detector array. The most significant decrease in background occurred with the two-detector array (28 to 40%), while background was increased by 18-43% for the four-detector array. Arrays consisting of three and four 80 x 20 mm broad energy germanium detectors resulted in increased relative sensitivity at all photon energies. The three-detector array provided the greatest sensitivity at photon energies below 344 keV. The four-detector array provided slightly better measurement sensitivity at photon energies greater than 344 keV. The two 80 x 20 mm detector array provided sensitivity unexpectedly comparable to the standard array. Monte Carlo predictions on how size affects counting efficiency and minimum detectable amount agreed well with the experimental results. From the Monte Carlo predictions, the effect of detector thickness on counting efficiency was unimportant at photon energies up to 60 keV and independent of detector diameter. At higher photon energies for both detector diameters, the counting efficiency decreased as the thickness decreased. The values of minimum detectable amount for the 70-mm and 80 mm diameter detectors did not differ by more than 15% at 17 keV or 20% at 60 keV when compared to detectors of equivalent thickness. Minimum detectable amount increased slightly at 17 keV and rose by approximately 52% at 660 keV, with decreases in thickness from 30 mm to 15 mm. PMID- 11725892 TI - Hiroshima neutron fluence on a glass button from near ground zero. AB - A decorative glass button that was uncovered at a location that is 190 +/- 15 m from directly beneath the atomic explosion at Hiroshima on 6 August 1945 has been scanned for induced fission tracks produced mostly by the thermal neutrons from the bomb due to interactions with the trace uranium that is normally present in silicate glasses. In surveying 4.14 cm2 at 500x magnification, 28 tracks were seen. From a calibration irradiation in a nuclear reactor we infer that the neutron fluence in 1945 was 5.7(+/-1.1) x 10(11) cm(-2); and, allowing for shielding by the structure in which the button was probably located, the free-air (i.e., outside) value is estimated as 1.5(+/-0.5) x 10(12) cm(-2). A limit has been placed on possible fading of the radiation-damage tracks that could increase the fluence by at most a factor of 1.27. The values bracket the calculated value of 9 x 10(11) given in DS86 but are higher than the 3.6 x 10(11) inferred from induced radionuclides for the distance given. The difference is, however, within the observed variability of the two types of results. PMID- 11725893 TI - 222Rn concentration in ground waters of coastal Karnataka and Kaiga of south west coast of India. AB - Groundwater (open well and bore well) samples from various locations of coastal Karnataka and Kaiga have been investigated for their 222Rn concentrations by emanometry method. The concentration of 222Rn in open well water was found to vary in the range 0.14-25.4 Bq L(-1) with a median value of 3.74 Bq L(-1) and that in bore well water in the range of 0.22-197.0 Bq L(-1) with a median value of 5.75 Bq L(-1). From the measured concentrations the effective doses for the population of the region were estimated. The effective dose was found to vary from 0.09 microSv y(-1) to 204.2 microSv y(-1) for open well water consumers and from 0.2 microSv y(-1) to 1586.9 microSv y(-1) for bore well water consumers. The results of the present study are also compared with the literature values and discussed. PMID- 11725894 TI - Radon health spas. PMID- 11725895 TI - Public involvement in science and decision making? PMID- 11725896 TI - Revised "joint statement" clarifies relationships between midwives and physician collaborators. PMID- 11725897 TI - Vaginal delivery after cesarean section: is the risk acceptable? AB - Recent studies and commentary have questioned the safety of vaginal births after cesarean sections (VBACs). The history of VBACs is reviewed and a framework to continue the safe practice of trials of labor in women with a prior uterine scar is presented. PMID- 11725898 TI - Measuring outcomes of midwifery care: development of an instrument to assess optimality. AB - Research on the outcomes of midwifery care is hampered by the lack of appropriate instruments that measure both process and outcomes of care in lower risk women. This article describes an effort to adapt an existing measurement instrument focused on the optimal outcomes of care (The Optimality Index-US) to reflect the contemporary style of U.S.-based nurse-midwifery practice. Evidence for content validity of the instrument was derived from literature reports of randomized clinical trials, synthetic reviews, and the clinical consensus of professional reviewers. Eleven perinatal health professionals and consumers, representing disciplines of obstetrics and gynecology, midwifery, epidemiology, and neonatology reviewed the instrument. The instrument was then applied to an existing data set of women who intended to give birth at home (N = 1,286 women) to determine its utility in measuring events in the process and outcome of perinatal health care as managed by nurse-midwives. Results suggest that the tool holds promise for use in outcomes studies of U.S. perinatal care. Further testing of the instrument among diverse multicultural population groups, with various providers, and in diverse birth settings is warranted. PMID- 11725899 TI - Women's health care: a new paradigm for the 21st century. AB - Oppression based on gender exists in all aspects of women's lives and transcends contemporary cultures, economic systems, and even health care services. Radical feminism provides an alternative philosophic framework of health care that is based on a women-centered viewpoint, with the experiences of women as its unifying philosophy. Midwifery is a means to apply this new philosophic approach to the health care of women. A partnership between midwifery and feminist philosophy will allow women's voices to be heard, while guiding research in women's health care in new directions, and illuminating new approaches to current health problems. The new millennium provides an opportunity to explore an alternative framework and philosophy that will change the current paradigm of women's health care. PMID- 11725900 TI - Screening for hypoglycemia in healthy term neonates: effects on breastfeeding. AB - Transient hypoglycemia in the early neonatal period is a common adaptive phenomenon as the newborn changes from the fetal state of continuous transplacental glucose consumption to intermittent nutrient supply following cessation of maternal nutrition at birth. Research has demonstrated that in the term, healthy newborn, this dynamic process is self-limiting and is not considered pathologic. The American Academy of Pediatrics and the World Health Organization recommend that neonatal blood glucose screening be reserved for newborns who are at risk or symptomatic and conclude that universal hypoglycemia screening is inappropriate, unnecessary, and potentially harmful. Nevertheless, many hospital nurseries continue the clinical practice of routine early glucose screening on healthy, term newborns. This results in the misidentification of neonates captured while experiencing the normal, self-correcting physiologic blood glucose nadir who are then diagnosed with pathologic neonatal hypoglycemia. Subsequent to this misdiagnosis, further surveillance and unnecessary, aggressive treatment interventions will follow that are potentially harmful to the successful establishment of positive maternal-infant interactions and the breastfeeding experience. Research studies indicate that routine hypoglycemia screens, treatments, and interventions in the healthy infant are not evidence based and result in a serious disruption of the initiation process and duration patterns of lactation. Using the perspective of the theory of technology dependency, this inquiry explores the potential adverse sequelae of inappropriate glucose screening in the healthy breastfeeding newborn and describes selected outcome variables including: 1) the consequences of early maternal-infant separation, 2) the influence of early formula supplementation on breastfeeding discontinuance rates, 3) the effect of separation and supplementation on the onset of lactogenesis, and 4) the impact of hospital staff and provider recommendations of formula supplementation on maternal confidence to independently nurture her baby. PMID- 11725901 TI - Women and smoking. PMID- 11725902 TI - The national strategy for suicide prevention. PMID- 11725903 TI - The midwife as first assistant for cesarean section. AB - Across the United States, midwives have expanded their role to include serving as first assistants at cesarean sections. An American College of Nurse-Midwives (ACNM) Position Statement adopted in 1998 recognizes the practice as a "frequently-performed advanced midwifery practice skill." Workshops have been offered nationally in 1997, 1998, 2000, and 2001 as well as locally in some states to educate and guide the midwife in completing the didactic and clinical preparation. Yet, there is a dearth of published literature on the subject. This article reviews the evolution of the role from its origins in perioperative nursing, including the requirements for the Registered Nurse First Assistant as established by the Association of peri-Operative Registered Nurses that are referenced in the regulations of several states in regard to the practice of midwives as surgical assistants. The authors report the results of a survey of state regulatory agencies that reveals a wide variation in laws, regulations, and interpretations ranging from statutory acceptance to prohibition with suggestions on how to effect needed change. This article also includes a process for credentialing that is consistent with the ACNM "Guidelines for the Incorporation of New Procedures into Midwifery Practice." The importance of documentation of the educational and credentialing process from a medico-legal perspective is stressed. PMID- 11725904 TI - The 1999-2000 ACC task analysis of nurse-midwifery/midwifery practice: a consideration of the concept of professional issues. AB - The American College of Nurse-Midwives (ACNM) Certification Council periodically conducts a task analysis study as evidence supporting the content validity of the national certification examination in nurse-midwifery and midwifery. The purpose of this article is to report findings related to the examination of the relationship between professional issues and safe beginning-level midwifery as measured by the 1999-2000 Task Analysis of American Nurse Midwifery and Midwifery Practice. Study findings suggest that newly certified midwives place strong emphasis on the importance of tasks related to the ACNM "Hallmarks of Midwifery," which characterize the art and science of the profession: these include tasks dealing with health promotion and cultural competency. The beginning midwives, however, gave consistently low ratings to tasks related to ACNM "Core Competencies" that mirror the professional responsibilities of midwives; these include tasks related to the history of midwifery, research, or health policy. The study has implications for nurse-midwifery/midwifery educators, experienced midwifery mentors, and other persons interested in reinforcing the relevance of these important professional issues to the new midwife. PMID- 11725905 TI - Clinical bulletin no. 5--June 2001. Endometrial biopsy. PMID- 11725906 TI - Betty Watts Carrington, CNM, EdD, FACNM. Recipient of the Hattie Hemschemeyer Award for 2001. PMID- 11725907 TI - Effect of mycophenolate mofetil on the progression of adriamycin nephropathy. AB - In order to assess the effects of mycophenolate mofetil (MMF) on the development of adriamycin-induced nephropathy, the development of this nephropathy in rats treated with MMF was compared to that in non-treated animals (group ADR + V) over 28 weeks. At weeks 8, 16 and 20, 24-h proteinuria of the treated group statistically differed from that of the non-treated group. However, no significant difference in proteinuria was observed thereafter between the groups. At the end of the experiment, there was no significant difference between both groups regarding the frequency of glomerular lesion (Group ADR + V: Md = 35%, P25 = 20%, P75 = 68%; Group ADR + MMF: Md = 27%, P25 = 9.5%, P75 = 54%); tubulointerstitial lesion index (Group ADR + V: Md = 7, P25 = 1.5, P75 = 9; Group ADR + MMF: Md = 8, P25 = 2, P75 = 9); glomerulosclerosis area (group ADR + V = 2779 microm2, P25 = 751.8 microm2, P75 = 3115 microm2; Group ADR + MMF = 1147 microm2, P25 = 3969.7 microm2, P75 = 1560 microm2); and, interstitial fibrosis area (Group ADR + V: Md = 218200 microm2, P25 = 78670 microm2, P75 = 282700 microm2 group ADR + MMF: Md = 136000, P25 = 25010, P75 = 255800 microm2). In conclusion, MMF caused a temporary reduction in proteinuria but did not change the severity of the renal lesion observed after 28 weeks. PMID- 11725908 TI - Lipogenesis in experimental chronic renal failure in rats. AB - Hyperlipidemia is a common occurance in patients with chronic renal failure (CRF) and has been the subject of many clinical and experimental studies. Despite this, the role of lipogenesis in the development of hyperlipidemia is still obscure. The present study is based on a rat model of CRF involving a two-stage subtotal nephrectomy. In this study, we measured the activity of fatty acid synthase (FAS). This is the rate-limiting enzyme of lipogenesis and is present in liver and white adipose tissue (WAT). Using isotopic methods, we also determined the rate of lipogenesis in vivo in liver and WAT. In both liver and WAT, the results of the analyses were similar. In the uremic rats, there was a tendency for the FAS activity to rise. However, the difference was not statistically significant. Furthermore, there was no increase in the rate of lipogenesis in vivo in either tissue. In summary, the results of our study confirm the thesis that lipogenesis does not play a role in the development of hypertriglyceridemia seen in an experimental CRF in rats. PMID- 11725909 TI - Granulocyte macrophage colony stimulating factor (GM-CSF) induced sero-protection in end stage renal failure patients to hepatitis B in vaccine non-responders. AB - Hepatitis B (HB) virus infection is a major health problem in dialysis dependent end stage renal failure (ESRF) patients. The sero-conversion rate after recombinant HB vaccine in ESRF patients is poor. Adjuvants like Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) have been found to improve response rate to vaccines. This study was conducted to evaluate the efficacy of GM-CSF as an adjuvant to HB vaccine in ESRF patients who were non-responders to the usual three double dose vaccinations (primary non-responders). Fifty consecutive HBsAg negative and anti-HBs negative ESRF patients on hemodialysis over thirty months were prospectively included (Jan. 96-June 98). All received 40 microg of recombinant HB vaccine at 0, 1, 2 month interval. Anti-HBs titres were subsequently tested after four weeks of the third dose. There were 19 (38%) primary non-responders (antiHBs negative). Twelve (Group I) of primary non responders were given an additional dose of HB vaccine with 300 microg (5-6 microg/kg) of GM-CSF (Leucomax) and the remaining seven (Group II) received only an additional dose of HB vaccine. Anti-HBs was determined by Abbott's ELISA kit, and titre above 10 mIU/mL was considered as protective. In Group I, sero protective titres were obtained in 11 out of 12 (91.6%) patients, whereas in Group II none of the patients achieved sero-protection (p < 0.001). The sero conversion rate improved from initial 62% (31/50) to overall 84% (42/150) after the use of GM-CSF. There were no adverse events noted with the use of GM-CSF. At one year, 24 out of 32 (75%) who were sero-protected earlier continued to remain sero-protected. This study indicates that GM-CSF is a potent HB vaccine adjuvant for sero-conversion in primary non-responders. PMID- 11725910 TI - A potential role for immune activation in hemodialysis hypotension. AB - The necessary exposure of blood to biomembranes during hemodialysis has been viewed by many as an immunogenic challenge leading to an acute phase response. In this study we examined the relationship between hemodialysis-induced immune activation and intradialytic hypotension, using the acute phase reactant serum C reactive protein (CRP) as a surrogate for immunogenic activation. The maximum percent change in mean arterial pressure (MAP) was found to correlate significantly with CRP (r = 0.67, p < 0.05) in nine consecutive patients with a history of symptomatic hypotension during hemodialysis. In contrast, no correlation was found between CRP and maximum percent change in MAP in eight consecutive hemodialysis patients without intradialytic hypotension. Since interleukin-6 (IL-6) is a major regulator of CRP, the relationship between these two proteins was examined. Plasma IL-6 levels were found to correlate both with CRP (r = 0.67, p < 0.05) and with mean maximum percent change in MAP (r = 0.70, p < 0.05) in hemodialysis patients with a prior history of hypotension. IL-6 levels did not correlate with CRP or blood pressure in the hemodynamically stable patients. The results suggest that immune activation working through IL-6, CRP and other cytokines may play a role in the pathogenesis of hemodialysis hypotension in some patients. PMID- 11725911 TI - Enalapril acts through release of nitric oxide in patients with essential hypertension. AB - BACKGROUND: Endothelial dysfunction has been proposed as an etiological agent in the pathogenesis of essential hypertension. Amongst the various antihypertensive drugs, angiotensin converting enzyme inhibitors (ACEI) have been implicated in modifying the vascular endothelium by the release of mediators that include bradykinin, nitric oxide, prostaglandins and thromboxane A2. MATERIALS AND METHODS: To study the mechanism of action of enalapril, an ACEI, serum reactive nitrite intermediates (RNI) and citrulline, by products of nitric oxide metabolism were measured before and after treatment with enalapril in 25 consecutive patients of essential hypertension. RESULTS: Following treatment serum RNI intermediate increased from a pretreatment value of 164.5 +/- 20.2 nmol/mL to a post treatment value of 266.9 +/- 47.3 nmol/mL (p < 0.05), however there was no significant change in the levels of citrulline (p > 0.1). There was no significant correlation between the severity of hypertension and serum RNI. Serum RNI levels were lower in the postmenopausal women but did not reach statistical significance. CONCLUSIONS: It is postulated that enalapril exhibits its antihypertensive property through release of nitric oxide. PMID- 11725912 TI - Amyloid goiter in Familial Mediterranean Fever (FMF): a clinicopathologic study of 10 cases. AB - FMF Amyloidosis is an important etiological factor of end stage renal disease (ESRD) in Mediterranean Countries. Apart from major target organs as cardiovascular, respiratory and gastrointestinal system, endocrine organs can also be involved. We planned to investigate the thyroid involvement in our amyloidosis group. The aim of this study was to determine clinical characteristics of amyloid goiter in FMF patients and the abnormalities of thyroid function, as well as to identify pathologic characteristics. Twenty-two hemodialysis patients (mean age 34.1 +/- 14 years, range 17-68) whose ESRD secondary to FMF amyloidosis were evaluated with physical examination, serum levels of thyroid hormones, ultrasound examination of thyroid glands, thyroid syntigraphic studies. Goiter was found in 10 patients (4 male, 6 female) having enlarged neck mass (mean age 35 +/- 14 years, range 23-64). The serum levels of thyroid hormones and TSH were normal in 4 patients. Other four cases had euthyroid sick syndrome. Only one patient developed tender enlarged neck mass with subacute thyroiditis symptoms and one had primary hyperthyroidism. Ultrasound examination showed; hypoecoic nodules in 6 patients diffuse multinodular enlargement in 4 patients. Thyroid syntigraphic studies revealed hypoactive nodules in 7 patients and hyperactive nodules in 3 patients. After the laboratory tests were completed, in 10 patients diagnosis were made with fine needle aspiration biopsy. Of 10 patients 5 underwent subtotal thyroidectomy. Immunohistochemical evaluation demonstrated the presence of amyloid AA immunoreactivity in all cases. In conclusion fine needle aspiration from the thyroid when enlarged is useful in the diagnosis of suspected amyloidosis, especially since it is a safe, easily performed procedure. With the help of amyloid goiter diagnosis the patient's prognosis on hemodialysis and with renal transplantation can be predicted. Amyloid goiter must be searched in hemodialysis patients especially in Mediterranean Countries. PMID- 11725913 TI - Coronary artery disease among diabetic and non-diabetic patients with end stage renal disease. AB - Cardiovascular disease is the major cause of death among patients with end stage renal disease and accounts for about half the deaths among the dialysis population. Several researchers have reported a high prevalence of coronary artery disease among diabetic patients with renal failure and coronary arteriography is often considered an integral part of the pre-transplant evaluation of diabetic patients with end stage renal disease. However, very few reports have addressed the question of coronary disease in non-diabetic patients, and the pattern and prevalence of coronary artery disease in non-diabetic patients with end stage renal disease are not well defined. We evaluated the clinical and coronary angiographic findings in 158 consecutive patients (84 diabetic and 74 non-diabetic) with end stage renal disease. The coronary arteries were divided into 16 segments and each segment was analyzed for the presence of coronary disease, which was defined as the presence of > or = 50% luminal diameter stenosis. Diabetic patients had more adverse risk factors for coronary artery disease, yet there was no significant difference in the prevalence of coronary artery disease between the diabetic and non-diabetic patients (67% vs. 55%, p = 0.15), or in the number of affected coronary artery segments (2.0 vs. 1.4, p = 0.05). Triple vessel coronary artery disease was however, significantly more common among the diabetic subjects (27% vs. 12%, p = 0.005). Non-diabetic patients with end stage renal disease also have a high prevalence of coronary artery disease and may merit as careful investigation of their coronary status as their diabetic counterparts. PMID- 11725914 TI - Complication of oliguric acute renal failure in patients treated with low molecular weight dextran. AB - Acute renal failure (ARF) is a well-documented but infrequent complication in patients treated with low-molecular weight dextran (LMWD). We herein report 3 cases of oliguric ARF following the administration of dextran-40. One case developed ARF totally after 1.200 g of LMWD administration. In contrast, two cases having increased serum creatinine developed oliguria despite the acceptable therapeutic doses (totally 450 and 650 g). Contrast media was also co administered in these patients. Plasma exchange (PE), double filtration plasmapheresis (DFPP), or continuous hemodiafiltration (CHDF) but not hemodialysis (HD) reduced circulating dextran concentrations by 35-44% during a single session. All patients completely recovered from ARF by 14-32 days after the treatment. Our cases suggested that radiocontrast could predispose to the development of LMWD-induced ARF especially in patients having pre-existing renal dysfunction. In addition, PE, DFPP and CHDF afforded a beneficial effect for removing accumulated LMWD from the circulation. PMID- 11725915 TI - The effect of Losartan on insulin resistance and beta cell function in chronic hemodialysis patients. AB - Insulin resistance (IR) is prevalent in hemodialysis patients. IR and hyperinsulinemia have an important role in the development of atherosclerosis, which is the most common cause of morbidity and mortality in hemodialysis patients. Thus, antihypertensive drugs that lower IR, may have an additional beneficial effect in the treatment of cardiovascular diseases in these patients. In this preliminary study we examined the effect of Losartan (an angiotensin II receptor antagonist) treatment on IR and beta cell function in five hypertensive non-diabetic chronic hemodialysis patients. All other known causes of IR in end stage renal failure were excluded. After a washout period of two weeks, Losartan 50 mg, was administered for 6 weeks. Fasting blood glucose (FBG) and insulin levels were measured before and after the treatment IR and beta cell function were calculated using the "homeostasis model assessment"-HOMA. Systolic and diastolic blood pressure (BP) have not changed significantly throughout the study. FBG increased significantly from 76 mg/dL +/- 1 to 89 mg/dL +/- 4 (p < 0.01), however, insulin levels have not changed significantly. Calculated IR values did not show a difference, but calculated beta cell function decreased significantly after Losartan treatment from 291% +/- 50 to 146% +/- 10, (p < 0.016). These preliminary results suggest that in chronic hemodialysis hypertensive non-diabetic patients short treatment with Losartan has deleterious effect on glucose homeostasis mediated via a decrease in beta cell function. PMID- 11725916 TI - Tubular cell lesion, albuminuria, and renal albumin handling in rats treated with adriamycin. AB - Several lines of evidence have suggested that renal handling of proteins in rats with several nephropathies may contribute to the tubulointerstitial damage observed in these animals. It has been suggested that proteins filtered by the glomeruli may be toxic for tubule cells. The aim of this study was to investigate the relationship between albuminuria and tubular lesions observed in rats during the first two weeks after treatment with adriamycin (AD). Thirty female Wistar rats were injected intravenously with adriamycin at the dose of 3.5 (17 rats) or 5mg/kg body weight (13 rats), and 7 were injected with 0.15 M NaCl (control group). Seven days later, we replaced drinking water with a 0.10 M sodium bicarbonate solution for 6 of the animals injected with 5 mg/kg adriamycin (group AD-B). Urine samples were collected before and 7 and 15 days after treatment to quantify albumin. The rats were killed 7 and 18 days after the injections, and the kidneys removed for immunohistochemical study. We observed a significant increase in urinary albumin excretion 15 days after AD injection (3.5 mg/kg), but not 7 days after AD. However, in the animals injected with 5.0 mg/kg AD (group AD 5) the increase in albuminuria was observed as early as on day 7. The immunohistochemical studies showed increased vimentin and albumin immunoreaction in the tubular cells of the renal cortex from the kidneys of rats injected with 3.5 mg/kg (group AD-3) only 18 days after treatment (p < 0.05), whereas in the animals treated with 5 mg/kg AD these immunohistochemical alterations were more intense. However, treatment with sodium bicarbonate attenuated the tubular lesions and reduced albumin reabsorption in adriamycin-treated rats. In conclusion, these experiments showed a relationship between albuminuria and tubular lesions in adriamycin-treated rats. PMID- 11725917 TI - The HELLP syndrome, a prospective study. AB - OBJECTIVE: We undertook this study to evaluate the incidence and outcome of HELLP in Irish patients. In addition, duration and trends of the abnormal laboratory results were studied. STUDY DESIGN: This prospective observational study screened 12068 pregnant women between January 1995 and March 1997. Any pregnant woman with hypertension, proteinuria, thrombocytopenia or anemia was monitored for hemolysis and elevated liver transaminases, from the time of recruitment till six weeks postpartum or resolution. RESULTS: Thirteen of 12068 pregnant women (0.11%) developed HELLP. All had hypertension and 84.6% had proteinuria. Delivery was the only factor found to terminate the syndrome. Acute renal dysfunction was noted in 53.8% but none required dialysis. Laboratory parameters stabilized by the sixth postpartum day. Fetal mortality was 1 out of 14. There were no maternal deaths. CONCLUSIONS: HELLP syndrome is a rare but potentially serious complication of pregnancy. Correlation with laboratory data and early intervention are vital in achieving a favorable outcome for both mother and fetus. PMID- 11725918 TI - Anticipation of end stage renal disease in patients with autosomal dominant polycystic kidney disease in successive generations. AB - Autosomal dominant polycystic kidney disease is a disorder, which is inherited in 50% of offspring, irrelevant the sex and it has a variable clinical expressivity. Initially it was noticed that the clinical expression was interfamilial, but some studies found out that it was different. The aim of this study was to evaluate the age of onset of end-stage renal disease (ESRD) in affected parents in comparison with their offspring in successive generations. We studied 60 families of patients with autosomal dominant polycystic kidney disease (ADPKD). The diagnosis was done by echo criteria and we included only the patients for whom we knew precisely the onset of ESRD (affected parent and offspring), the sex of the parent who suffered from the disease, and offspring. We found out that the ESRD in ADPKD appears at the same age in affected parents and offspring (49,3 +/- 7,9 Vs 51,8 +/- 9,2, p = NS) irrelevant of the sex of the offspring. Patients with paternal inheritance (n = 38) were diagnosed to have ESRD earlier than their affected parents (47,9 +/- 8,3 Vs 52,2 +/- 9,2 p < 0,05), but patients with maternal inheritance had no difference (n = 22) (51,9 +/- 6,8 Vs 51,2 +/- 9,4, p = NS). In all the patients (60 couples) the survival rate was the same between affected parents and offspring (p = NS, Kaplan-Meier test), but significant differences were noticed between offspring with paternal inheritance in comparison with their parents (p < 0,05). In conclusion, we have detected that the onset of ESRD between patients with ADPKD in successive generations: a) Occurs in offspring as in their ancestors, b) anticipation was observed in 55% of couples, c) the sex of offspring does not have any relation with the renal death and d) the ESRD in patients with paternal inheritance occurs earlier in offspring than in their ancestors but not with maternal. PMID- 11725919 TI - An enormous abdominal mass associated with acute renal failure. AB - We report a 67-year-old man with acute uric acid nephropathy, secondary to spontaneous tumor lysis syndrome, that presented itself as a huge intra-abdominal tumor that led to acute renal failure, hyperuricemia, and azotemia. Initial finding of hydronephrosis detected by ultrasonography led us to believe that the azotemia and decreasing amount of urine resulted from obstructive uropathy, a common complication of malignancy, caused by either a direct renal invasion or a urinary outflow tract compression because of a tumor mass effect. However, clinical observations and the response to therapeutic intervention confirmed the diagnosis of spontaneous tumor lysis syndrome, which is a rare cause of acute uric acid nephropathy. PMID- 11725920 TI - Classic heat stroke in a case of simple hypohydrosis with "bad prognostic indicators" but a remarkable recovery. AB - Heat stroke occurs in the desert area of Saudi Arabia quite frequently and manifest in different patterns including coagulopathy. Frequently encountered complications include renal or hepatic failure, rhabdomyolysis, acute respiratory distress syndrome (ARDS), disseminated intravascular coagulation (DIC), and seizure. Not all of these complications usually occur in the same patient, in case it occurs the mortality reported is significantly high. We describe a case of heat stroke that had nearly all the known complications of heat stroke but recovered from all, except minor neurological deficit in the form of dysarthria and exaggerated deep reflexes. PMID- 11725921 TI - Chronic interstitial nephritis following parenteral copper sulfate poisoning. AB - A 21-year-old male patient was admitted with acute renal failure and intravascular hemolysis following suicidal parenteral copper sulfate poisoning. He developed metabolic acidosis and septicemia; and was treated with intensive hemodialysis, blood transfusions and antibiotics. After remaining anuric for 4 weeks, his urine output gradually increased. However his renal functions improved only partially. Renal biopsy done 8 weeks after the episode showed chronic tubulo interstitial nephritis (CIN). This is the first reported case showing CIN following acute copper sulfate intoxication. PMID- 11725922 TI - Thrombotic thrombocytopenic purpura in a case of scleroderma renal crisis treated with twice-daily therapeutic plasma exchange. AB - We report the rare complication of thrombotic thrombocytopenic purpura (TTP) in an elderly patient with the uncommon scleroderma renal crisis (SRC) at the advanced age of 85 years. Initially, she was treated by standard daily one and a half plasma volume therapeutic plasma exchange (TPE), in combination with steroid therapy. Due to an unsatisfactory clinical response, she was consequently, treated by an intensive twice-daily one and a half plasma volume TPE regimen. Although there was an increase in the platelet count, a 7-day course of the intensive regimen did not elicit further propitious laboratory test results or a clinical improvement and the patient expired secondary to the pulmonary and cardiac conditions related to SRC. Intensive TPE administration has been advocated in few specific patients who are refractory to standard daily treatment. The unfavorable outcome in this case may be attributed to the multiple complications associated with her primary disorder. In conclusion, we observed intensive twice-daily TPE to increase temporarily the platelet count, but not to be of benefit in a patient with SRC and multiple organ failure. The two TPE regimens and outcomes are compared and the pathogenesis of TTP and scleroderma are discussed. PMID- 11725923 TI - Lymphatic tumor emboli of perirenal fat in patient with nephrotic syndrome receiving renal biopsy, ultimately revealed gastric adenocarcinoma with membranous glomerulonephritis. AB - The association of malignancy with nephrotic syndrome and renal histopathologic abnormalities is well documented. Paraneoplastic proteinuria caused by membranous glomerulonephritis usually is made simultaneously with the diagnosis of a malignant tumor, or the two conditions are diagnosed within a year of each other. We reported a patient who presented with nephrotic syndrome initially. Incidentally, in kidney specimens, pathologic findings showed perirenal fatty tissue with malignancy tumor emboli in lymphatics. Thereafter, gastric adenocarcinoma was diagnosed by gastrointestinal panendoscopy with gastric biopsy under impression of malignancy associated with glomerulonephritis. Patient died of complications of malignancy-related disseminated intravascular coagulation without chemotherapy after confirming diagnosis was made three months later. PMID- 11725924 TI - Reduced activities of divalent cation activated ATP-ase and 5'-nucleotidase in glycerol induced acute renal failure. PMID- 11725925 TI - Does angiotensin II receptor antagonist Losartan suppress drinking behavior in hemodialysis patients? PMID- 11725926 TI - Evaluation of tests based on individual versus population modeling to compare dissolution curves. AB - This study is aimed at comparing dissolution curves obtained for several batches of sustained release formulations of octreotide, a somatostatin analog used in acromegaly. We evaluated four tests to compare the parameters of two series of curves: a Mann-Whitney-Wilcoxon test comparing individual estimates; a likelihood ratio test after population analysis (NONMEM), and two Wald tests using population parameters estimated by two-stage methods. The four approaches were evaluated by simulations. The Mann-Whitney-Wilcoxon test and the population approach had adequate type-I error, and the latter provided the highest power under the alternatives simulated. The two-stage method including the uncertainty on individual estimates was disappointing. PMID- 11725927 TI - Properties of multiple intersection-union tests for multiple endpoints in combination therapy trials. AB - We consider intersection-union tests involving multiple endpoints in a combination drug trial, for which we control the familywise error rate in the strong sense using closed testing methods. Bonferroni-Holm, Simes-Hommel, and Resampling-Based methods all are considered in this context. Familywise error rate control heuristics are developed and evaluated using a simulation study that is specifically tailored to the intersection-union setting. Both Resampling-Based and Simes-Hommel uniformly outperform the Bonferroni-Holm. Using simulations of power, the choice of Simes-Hommel versus Resampling-Based is seen to depend on the particular alternative of interest. Because it is simpler and has generally good power, we recommend the Simes-Hommel intersection-union tests. The techniques are illustrated using real data from a clinical trial to evaluate a combination asthma therapy. PMID- 11725928 TI - A measure of association between two sets of multivariate responses. AB - Let X and Y denote two distinct sets of multivariate responses, each of which can be a mix of continuous and ordinal data. Suppose that it is of interest to quantify the overall strength of association between X and Y. Our two approaches to this problem are to construct correlation coefficients similar to Kendall's tau in the bivariate case. The first approach uses the average concordance of each pair of individuals, where the average concordance is obtained across all possible pairings of variables between the two sets, as the kernel of the U statistic. Then the overall correlation between the two sets of multivariate data is expressed in terms of the expected value of the kernel. The second approach estimates pairwise Kendall's taus between the two sets, then uses a summary measure to quantify the overall association. One option for the summary measure is to take the average of taus, and another option is to take the distance of taus. The two approaches yield the same results when the summary measure of the latter is the average. However, using the distance of taus can sometimes detect relationships not seen in the average. We illustrate the proposed methods using data from a national asthma clinical trial in which the first set of variables comprises 12 binary responses of skin allergen tests and the second set of variables comprises three continuous outcomes of pulmonary function and one clinical outcome. PMID- 11725929 TI - Design and analysis of trials with quality of life as an outcome: a practical guide. AB - Health Related Quality of Life (HRQoL) measures are becoming more frequently used in clinical trials, as both primary and secondary endpoints. Investigators are now asking statisticians for advice on how to plan (e.g., sample size) and analyze studies using HRQoL measures. HRQoL measures such as the SF-36 are usually measured on an ordered categorical (ordinal) scale. In the designing stages and when analyzing, the scales are often scored and the scores treated as if they were continuous and normally distributed. However the ordinal scaling of HRQoL measures leads to problems in determining sample size, and conventional parametric methods of estimation and hypothesis testing may not be appropriate for such outcomes. We present practical guidelines for the design and analysis of trials with HRQoL measures as outcomes. We used conventional statistical methods (i.e., t-tests and multiple regression), various ordinal regression models (proportional odds, continuation ratio, polytomous and stereotype) and bootstrap methods to analyze an HRQoL dataset. To illustrate the various methods we used HRQoL data on the SF-36 Role Limitations Emotional dimension for two groups of patients with leg ulcers. The bootstrap, t-test, and multiple regression methods gave similar results. The various ordinal regression models also gave similar results. If the HRQoL measure has a large number of ordered categories, most of which are occupied, and the underlying scale really is continuous but measured imperfectly by an instrument with a limited number of discrete values, then an informal rule of thumb is that this discrete scale should be treated as continuous if it has seven or more categories and as ordinal otherwise. PMID- 11725930 TI - How to assess the relative range of two consecutive laboratory measurements to monitor patients. AB - An integral part of routine health checkups involves laboratory measurements on various analytes in the blood. It is then common to compare the value of two consecutive measurements sampled at different times from the same patient. A "significant" change requires an action (additional sample and/or clinical action). The current rule is to check whether the relative range of measurement is larger than a certain critical threshold. This rule should guarantee a specified confidence level (e.g., 95%), but its derivation was based on an approximation. We derive the exact distribution and show that it is related to the doubly noncentral F distribution. The currently used threshold values are compared with the exact ones, and some limited power calculations are presented to detect changes in the patient condition. PMID- 11725931 TI - Testing dose-response relationships with a priori unknown, possibly nonmonotone shapes. AB - Usually, a monotone dose-response dependence can be assumed for the simultaneous comparison of increasing levels of a certain drug. However, sometimes a reversal of the dose-response curve is likely to occur at the higher doses. We investigate such violations of the monotonicity assumption. Adequate alternatives are discussed and the "protected trend alternative" is introduced. Together with the umbrella patterns described in the literature, we introduce new testing approaches for both alternatives. P-values/quantiles and power values/sample sizes are made numerically available and hence are readily computed. A short power study and the analysis of a data set from the literature demonstrate the improved behavior of the new methods. PMID- 11725932 TI - Robust estimators of the concordance correlation coefficient. AB - This paper proposes a generalized version of Lin' s concordance correlation coefficient for the agreement assessment of continuous data. Lin's coefficient evaluates the accuracy and precision between two measures, and is based on the expected value of the squared distance function. We generalize Lin's coefficient, apply alternative distance functions, and produce more robust versions of the concordance correlation coefficient. In this paper, we develop the asymptotic theory for this class of estimators, investigate small-sample properties via computer simulation, and demonstrate their use with two real data examples. PMID- 11725933 TI - Mixed or mixed up methods? PMID- 11725934 TI - Ethnographic analysis of everyday ethics in the care of nursing home residents with dementia: a taxonomy. AB - BACKGROUND: The concept of everyday ethics was used to emphasize the moral basis of ordinary issues of daily living affecting quality of life for nursing home residents with dementia. OBJECTIVES: To critically examine ethical issues of daily living affecting nursing home residents with dementia and to construct a descriptive taxonomy inductively derived from ethnographic fieldwork data. METHOD: Combined anthropological methods of participant observation and in-depth interviewing were used in the natural setting of a 147-bed, voluntary, not-for profit nursing home. Experiences of 30 residents, their family members, and nursing home staff were explored. In addition, the records of 10 ethics committee cases involving residents with dementia further enlarged the database. RESULTS: The taxonomy of everyday ethical issues includes the following four domains: (a) learning the limits of intervention; (b) tempering the culture of surveillance and restraint; (c) preserving the integrity of the individual; and (d) defining community norms and values. Each is representative of constellations of concerns that are grounded in the cultural and moral environment of the nursing home. CONCLUSIONS: Results highlight the challenges of recognizing the ethical in the ordinary, and of resolving everyday issues in ways that enhance quality of life for residents with dementia and those (family and staff) who care for them. PMID- 11725935 TI - Data mining methods find demographic predictors of preterm birth. AB - BACKGROUND: Preterm births in the United States increased from 11.0% to 11.4% between 1996 and 1997; they continue to be a complex healthcare problem in the United States. OBJECTIVE: The objective of this research was to compare traditional statistical methods with emerging new methods called data mining or knowledge discovery in databases in identifying accurate predictors of preterm births. METHOD: An ethnically diverse sample (N = 19,970) of pregnant women provided data (1,622 variables) for new methods of analysis. Preterm birth predictors were evaluated using traditional statistical and newer data mining analyses. RESULTS: Seven demographic variables (maternal age and binary coding for county of residence, education, marital status, payer source, race, and religion) yielded a .72 area under the curve using Receiving Operating Characteristic curves to test predictive accuracy. The addition of hundreds of other variables added only a .03 to the area under the curve. CONCLUSION: Similar results across data mining methods suggest that results are data-driven and not method-dependent, and that demographic variables offer a small set of parsimonious variables with reasonable accuracy in predicting preterm birth outcomes in a racially diverse population. PMID- 11725936 TI - Effects of maternal employment and prematurity on child outcomes in single parent families. AB - BACKGROUND: Effects of maternal employment for preschool children vary based on specific characteristics of the mother's employment, the family's economic status, and the mother's attitudes about employment. However, there is limited research on a growing group of children at developmental risk-those born prematurely and living in a single-parent family. OBJECTIVE: To examine the effects of maternal employment and prematurity on child cognition and behavior in single-parent families. METHODS: Sixty preterm and 61 full-term preschool children were recruited through NICU admission records and birth records. Data were collected with the Kaufmann Assessment Battery for Children, Peabody Picture Vocabulary Test, Child Behavior Checklist, Parenting Stress Index, and the Life History Calendar. RESULTS: Greater hours employed was related to higher achievement and mental processing scores only. Less discrepancy between actual and desired employment was related to higher achievement, mental processing, and language scores and lower behavior scores. Prematurity was not related to child cognitive and behavioral performance. Only the relationship between discrepancy and language remained after statistical control. CONCLUSIONS: The concerns about negative effects of maternal employment on young children may be overstated, especially in low-income, single-mother families. These findings are especially important in the context of welfare reform. PMID- 11725937 TI - Staffing and pattern of mechanical restraint use across a multiple hospital system. AB - BACKGROUND: In an effort to enhance patient safety in acute care settings, governmental and regulatory agencies have established initiatives aimed at limiting the use of mechanical restraints. Concurrently, hospital staffing levels are undergoing changes raising concerns about the impact these changes may have on restraint use. No studies to date have described the impact these two initiatives have had on restraint use in acute care hospitals. OBJECTIVES: To determine across a multiple hospital system: (a) the rates, frequencies, duration, and timing of restraint use, and (b) the relationship between restraint use and staffing. METHODS: This was a secondary analysis of prospective, observational data from a large outcomes database for 10 acute care hospitals. Monthly data were obtained from 94 patient care units for periods ranging from 1 12 months for a total of 566 cumulative months during 1999. RESULTS: The system restraint application duration rate (total restraint hours/total possible hours) was 2.8% (hospital ranges: 0.3-4.4%). More restraints were applied on night shifts (48.8%; n = 5,296) than on day (33.5%; n = 3,634) or evening shifts (17.7%; n = 1,926) (p < .0001) and most applied at midnight (31.7%; n = 3,441) followed by 0600-0900 (33.3%; n = 3,614). There was a weak positive relationship between staffing and restraint use (r = 0.276, p = .0001) at the system level and units with higher staffing levels also had higher baseline restraint use (p < .0001). CONCLUSIONS: Restraint frequency, duration, and timing may have been altered by recent initiatives, and there is beginning evidence that differences exist between community, rural, and tertiary hospitals. While there is a weak positive relationship between higher staffing and restraint use at the system and unit level, further exploration of the influence of other factors, specifically patient acuity, are in order. The finding of unit variability and consistent restraint application times provides a starting point for further quality initiatives or research interventions aimed at restraint reduction. PMID- 11725938 TI - Research design and subject characteristics predicting nonparticipation in a panel survey of older families with cancer. AB - BACKGROUND: While recognized that loss of subjects over time may introduce bias and complicate statistical analysis in panel studies, it is seldom acknowledged that sampling bias starts with subjects who are eligible but do not participate. OBJECTIVES: Community-based recruiters identified 1,948 subjects as eligible to participate in a descriptive panel survey of older families with cancer. Focusing on the time between identification of eligible subjects until contact for the first interview for consenting subjects, the purpose of this study was to determine if subject or research design factors predicted who was more prone to nonparticipation. METHOD: A multivariate model explored the simultaneous effects of subject and research design characteristics on nonparticipation. Demographic and cancer characteristics, as well as features of the study protocol, were used as predictors in a multinomial logit regression model to enable a three-way comparison between nonconsenters (n = 748), consenters who dropped out prior to data collection (n = 208), and consenters who participated in the intake interview (n = 992). RESULTS: Age and cancer diagnosis played roles in whether consent was obtained, whereas race did not affect consent but raised odds of drop out after consent. Powerful evidence emerged that design features, such as if a caregiver participated, recruitment staff roles, and payment to recruiters, affected the probability of subjects not participating or dropping out before being interviewed. CONCLUSIONS: Findings suggest that both subject and research design characteristics affect the likelihood of nonparticipation in a panel study of older cancer patients and family caregivers. Future research involving testing of strategies addressing access and accrual issues, research staff roles, reimbursements, and responsiveness to the needs of research personnel, ill participants, and family members is warranted. PMID- 11725939 TI - Recruitment and retention of community-dwelling, aging women in nursing studies. AB - BACKGROUND: Despite recent attention to women's lack of participation in health research, little nursing literature documents or describes recruitment and retention issues related to an increasingly large segment of the population, aging women who dwell in the community. OBJECTIVES: An analysis of a subset of published nursing research was conducted to identify: (a) demographics of samples that included community-dwelling, aging women; (b) recruitment and retention issues; and (c) the frequency and completeness with which each of these areas is reported. METHOD: An electronic database search was conducted for reports of funded nursing research that included women aged 65 and older that were published in a subset of English-language nursing journals between 1994 and June 1999. Research reports in which community-dwelling women were recruited for participation were selected for analysis (N = 50). RESULTS: Racial composition of the sample was included in 66% of reports; educational and employment status in 68% and 48%, respectively; and marital status in 64%. The majority of participants was White and married. It was impossible to determine the racial and marital status of the remainder of participants because of incomplete or ambiguous reporting. Recruitment and retention issues were addressed in only 23% of relevant reports, but specific recommendations can be derived from these reports to help other researchers. CONCLUSION: It is imperative for nurse researchers to include more information in research reports about their samples of community-dwelling, aging women and their successes and failures with recruitment and retention. This will help future researchers to study the population and will link nursing research to the larger social context of women's lives. PMID- 11725940 TI - Effects of a minimal intervention to increase physical activity in women: daily activity records. AB - BACKGROUND: Effective interventions to increase physical activity levels are critical in a nation where inactivity is a national public health problem. OBJECTIVE: This pilot study examined whether a minimal intervention (daily records of physical activity) increased activity levels in a community sample of working women. METHODS: In a longitudinal, pretest-posttest design, 49 working women were randomly assigned at the work site level to the control (n = 25) or intervention group (n = 24). At pretest and posttest, subjects completed self report questionnaires that measured psychological, social-environmental, physical activity, and demographic variables. Subjects in the intervention group kept daily records of their physical activities during the 12-week study, while those in the control group kept no records. In order to compare activity in the two groups, all subjects wore pedometers daily that recorded number of steps. RESULTS: There was a significant difference between groups in the pedometer values (mean number of daily steps) at the end of the study period (mean difference +/- SE 2147 +/- 636, p = .022) (2000 steps = approximately 1 mile). Multiple regression analysis showed that only the intervention (p = .003) was a significant predictor of the pedometer values. Hierarchical data analysis was used to account for the intra-class correlation of 0.48 within work site. CONCLUSION: Results from this sample of 49 women indicated that mean activity was greater in the intervention group compared to the control group. Recording daily activity is a cost-effective and acceptable intervention that may increase activity levels in women. However, more research is recommended to study the dual role of activity records as a data collection method as well as a potential intervention to increase physical activity. PMID- 11725941 TI - Conducting survey research at nursing conferences. AB - BACKGROUND: Conferences can provide large numbers of potential subjects in one location, yet there is little published guidance on how to collect data at a conference site. A computer search revealed no citations on this topic. This article outlines a systematic strategy to plan and perform research at conferences. OBJECTIVES: This article provides a step-by-step process to guide researchers in planning and conducting survey research at conferences. DISCUSSION: Initial components in planning data collection at a conference include making a timeline and getting permission. Detailed advanced planning involves specific strategies for attracting participants, and for distributing and collecting the questionnaires. Travel provisions and on-site logistical approaches are explained, followed by suggestions for post-conference activities. PMID- 11725942 TI - Handling missing data in nursing research with multiple imputation. AB - BACKGROUND: In the data analysis phase of research, missing values present a challenge to nurse investigators. Common approaches for addressing missing data generally include complete-case analysis, available-case analysis, and single value imputation methods. These methods have been the subject of increasing criticism with respect to their tendency to underestimate standard errors, overstate statistical significance, and introduce bias. OBJECTIVES: This article reviews the limitations of standard approaches for handling missing data, and suggests multiple imputation is a useful method for nursing research. METHOD: Secondary analysis was conducted to examine the effect of a public policy on the health of women using a data set that had a large degree and complex patterns of missing data. DISCUSSION: In the example, accommodation of the incomplete data was critical to making valid inferences; however, complete-case, available-case, or single imputation could not be defended as an adequate method for dealing with the missing data patterns. Alternative methods for dealing with incomplete data were sought, and a multiple imputation approach was selected given the missing data pattern. Nurse researchers confronting similar complex patterns of missing data may find multiple imputation a useful procedure for conducting data analysis and avoiding the bias associated with other methods of handling missing data. PMID- 11725943 TI - A catalytic subunit of the sugar beet protein kinase CK2 is induced by salt stress and increases NaCl tolerance in Saccharomyces cerevisiae. AB - Salinity is an important limiting factor in plant growth and development. We have cloned a catalytic subunit of the sugar beet protein kinase CK2 (BvCKA2) by functional expression in yeast of a NaCl-induced cDNA library. BvCKA2 was able to increase the yeast tolerance to NaCl and to functionally complement the cka1 cka2 yeast double mutant upon over-expression. Southern blot analysis indicated that, in sugar beet, the BCKA2 gene is a member of a multigene family. The mRNA levels of BvCKA2 were up-regulated in response to NaCl stress which suggests that protein kinase CK2 may be involved in the plant response to salt stress. PMID- 11725944 TI - The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression. AB - The cell wall polymer lignin is believed to be condensed by specific cell wall localized oxidoreductases. In many plants species, including poplar, the peroxidase-directed oxidation of the lignin analogue syringaldazine (SYR) has been localized to cells that undergo secondary wall formation, a process that includes lignification. As a first step to analyse the corresponding peroxidases. we have isolated previously two anionic isoenzymes (PXP 3-4 and PXP 5) from poplar xylem (Populus trichocarpa), which use SYR as a substrate. Here, we demonstrate that these enzymes are responsible for the visualized SYR oxidation in the developing xylem. The cDNA that corresponds to PXP 3-4 was isolated and the deduced protein was found closely related to the other SYR-oxidizing peroxidase PXP 5 (ca. 98% of identity). PXP 3-4 was expressed in a baculovirus expression system yielding high levels of active peroxidase (3 mg/l medium). The heterologously produced protein showed characteristics similar to those of the corresponding protein from poplar xylem (enzymatic properties, isoelectric point, and migration in a native gel). PXP 3-4 was expressed in the stem and in the root xylem. The data demonstrate that PXP 3-4 (and/or PXP 5) are present in differentiating xylem. supporting a function in secondary cell wall formation. PMID- 11725945 TI - Identical promoter elements are involved in regulation of the OPR1 gene by senescence and jasmonic acid in Arabidopsis. AB - Like other developmental processes, the terminal phase of leaf development, generally referred to as leaf senescence. regulates a subset of genes whose transcript abundances are increased during senescence. Jasmonic acid (JA), a plant growth regulator, also regulates the expression of subsets of genes in many aspects of plant growth and development, including leaf senescence. However, the underlying molecular mechanisms by which senescence and JA modulate gene expression are poorly understood. During an effort to isolate senescence associated genes, we identified an Arabidopsis enhancer trap line in which the reporter gene GUS is up-regulated by both senescence and JA. The T-DNA tagged gene was subsequently cloned using thermal asymmetric interlaced PCR (TAIL-PCR). This gene encodes a 12-oxo-phytodienoic acid-10,11-reductase (OPR1). Consistent with the GUS expression data, RNA gel blot analysis showed that OPR1 was indeed up-regulated by both senescence and JA. Promoter deletion analysis and linker scanning mutagenesis assays were employed to unveil the molecular bases of OPR1 regulation by senescence and JA. Two regulatory cis elements, namely JASE1 (5' CGTCAATGAA-3') and JASE2 (5'-CATACGTCGTCAA-3'), in the promoter region of the gene, were identified. While JASE2 contains a mixed A/C box-like motif, JASE1 represents a new motif without any signature sequence so far reported. Both elements were required for the up-regulation of OPR1 by leaf senescence and JA. suggesting that leaf senescence and JA may share a common molecular mechanism for modulating OPR1. PMID- 11725946 TI - Molecular identification and expression of the peroxidase responsible for the oxidative burst in French bean (Phaseolus vulgaris L.) and related members of the gene family. AB - Molecular characterization has been accomplished for five members of the peroxidase gene family in French bean. The most important of these, designated FBPI, corresponds to the isoform believed to be responsible for the apoplastic oxidative burst demonstrated by suspension-cultured cells in response to fungal elicitor. Identification was made by a complete match of six peptide sequences derived from the native protein to the translated sequence of the cDNA. Modelling of the surface structure in comparison with two other members of the peroxidase family did not reveal any unusual features which might account for its role in the oxidative burst. However, FBP1 when expressed in Pichia pastoris generated H2O2 using cysteine at pH 7.2, a specific property of the native protein when isolated from suspension-cultured cells. FBP1, together with other members of the family, were all induced in cell cultures by elicitor action although they all showed some expression in non-induced cultured cells. They were also expressed in all tissues examined with varying levels of intensity of detection in northern blots. This was confirmed by in situ hybridization and FBP1 expression was confirmed in tissues where it has been previously detected by immunolocalization methods. Assigning roles to individual peroxidases is an important goal and molecular identification of the oxidative burst peroxidase allows further exploration of the relative roles of the different systems involved in generating reactive oxygen species. PMID- 11725947 TI - Analysis of celery (Apium graveolens) mannitol dehydrogenase (Mtd) promoter regulation in Arabidopsis suggests roles for MTD in key environmental and metabolic responses. AB - Of the growing list of promising genes for plant improvement, some of the most versatile appear to be those involved in sugar alcohol metabolism. Mannitol, one of the best characterized sugar alcohols, is a significant photosynthetic product in many higher plants. The roles of mannitol as both a metabolite and an osmoprotectant in celery (Apium graveolens) are well documented. However, there is growing evidence that 'metabolites' can also have key roles in other environmental and developmental responses in plants. For instance, in addition to its other properties, mannitol is an antioxidant and may have significant roles in plant-pathogen interactions. The mannitol catabolic enzyme mannitol dehydrogenase (MTD) is a prime modulator of mannitol accumulation in plants. Because the complex regulation of MTD is central to the balanced integration of mannitol metabolism in celery, its study is crucial in clarifying the physiological role(s) of mannitol metabolism in environmental and metabolic responses. In this study we used transformed Arabidopsis to analyze the multiple environmental and metabolic responses of the Mtd promoter. Our data show that all previously described changes in Mtd RNA accumulation in celery cells mirrored changes in Mtd transcription in Arabidopsis. These include up-regulation by salicylic acid, hexokinase-mediated sugar down-regulation, and down-regulation by salt, osmotic stress and ABA. In contrast, the massive up-regulation of Mtd expression in the vascular tissues of salt-stressed Arabidopsis roots suggests a possible role for MTD in mannitol translocation and unloading and its interrelation with sugar metabolism. PMID- 11725948 TI - Organ specificity of a vacuolar Ca2+-binding protein RVCaB in radish and its expression under Ca2+-deficient conditions. AB - Radish vacuoles contain a new type of Ca2+-binding protein (RVCaB) with high capacity and low affinity for Ca2+. The protein is able to stimulate Ca2+ uptake into vacuoles, which is driven by Ca2+-ATPase and Ca2+/H+ antiporter. In the present study, we found that the level of RVCaB mRNA is high in seedling hypocotyls and mature taproots but low in young roots and mature leaves. The RVCaB protein was abundant in hypocotyls and taproots but absent in leaves. The levels of the transcript and protein of RVCaB in taproots were gradually increased during maturation. The level of RVCaB mRNA in seedling hypocotyls doubled within a few hours when the growth medium was changed from 10 mM CaCl2 to water, although the level was strongly suppressed in 100 mM CaCl2. This response of the RVCaB gene was specific to Ca2+ and did not occur with other ions including K+ and Mg2+. RVCaB functioning as a Ca2+-sequestering protein in taproot vacuoles to provide for the Ca2+ deficiency is discussed. PMID- 11725949 TI - The promoter of a basic PR1-like gene, AtPRB1, from Arabidopsis establishes an organ-specific expression pattern and responsiveness to ethylene and methyl jasmonate. AB - Antimicrobial proteins are a key feature underlying the deployment of both pre formed and inducible defence responses. Probably the most well characterised class are the pathogenesis-related (PR) proteins, which are found in both basic and acidic isoforms. Here we describe the isolation and characterisation of a gene, designated AtPRB1, encoding a basic PR1-like protein from Arabidopsis. This protein showed high amino acid sequence identity with basic and acidic PR1 proteins from other plant species, for example PRB1 from Nicotiana tabacum and PR1 from Brassica napus, at 64% and 78% identity respectively. A genomic DNA fragment containing 2345 bp upstream from the putative transcriptional start site was fused to the gene encoding the luciferase (LUC) gene from Photinus pyralis in order to test for promoter activity. The resulting construct was transformed into Arabidopsis accession Col-0 and analysis of LUC activity, using an ultra-low light imaging camera system, revealed that the AtPRB1 promoter established an exquisite organ-specific expression pattern. LUC activity was observed in flowers, stems and roots but not in leaf tissue. Superimposed upon this organ specific expression pattern was responsiveness, in root tissue, to ethylene and methyl jasmonate (MeJA), important cues during the establishment of plant disease resistance. In contrast, AtPRB1::LUC gene expression was repressed in response to salicylic acid treatment. Analysis of a limited series of AtPRB1 5'-promoter deletion mutants, identified a number of promoter regions important for both the establishment of organ-specific expression and responsiveness to ethylene and MeJA. While AtPRB1 gene expression was not induced in response to an avirulent isolate of Peronospora parasitica in leaf tissue, this gene may contribute to horizontal resistance in other tissues and/or to MeJA- and ethylene-dependent defence responses engaged against necrotrophic pathogens in root tissue. It is anticipated that transgenic plants containing AtPRB1-based promoter::reporter constructs will provide useful tools for the future dissection of the cognate signalling networks regulating the expression of this gene. PMID- 11725950 TI - Expression of the Pib rice-blast-resistance gene family is up-regulated by environmental conditions favouring infection and by chemical signals that trigger secondary plant defences. AB - The rice blast resistance gene Pib is a member of the nucleotide binding site (NBS) and leucine-rich repeat (LRR) class of plant disease resistance (R) genes and belongs to a small gene family. We describe here the isolation and characterization of a Pib homologue (PibH8), and extensive investigation of the expression of the Pib gene family (Pib, PibH8, HPibH8-1, HPibH8-2) under various environmental and chemical treatments. PibH8 shows 42% identity and 60% similarity to Pib and, like Pib, has a duplication of the kinase 1a, 2, and 3a motifs of the NBS region in the N-terminal half of the protein. Interestingly, genes of the Pib family exhibit a diurnal rhythm of expression. RNA gel blot analysis revealed that their expression was regulated dramatically by environmental signals. such as temperature, light and water availability. Their expression was also induced by chemical treatments, such as jasmonic acid, salicylic acid, ethylene and probenazole. Our findings suggest that expression of the Pib gene family is up-regulated by environmental conditions that would favour pathogen infection. This may reflect the evolution of anticipatory control of R gene expression. PMID- 11725953 TI - Basic and therapeutic relevance of endothelin-mediated regulation. AB - Three endothelin family peptides (endothelin-1, -2 and -3) exert an extremely potent and long-lasting vasoconstrictor action as well as other various actions through stimulating two subtypes of receptor (ETA and ETB). Vascular endothelial cells produce only endothelin-1. Although the pharmacological actions of exogenous endothelin-1 have been extensively analyzed, the physiological roles of endogenous endothelin-1 have long been obscure. Using potent and selective receptor antagonists, endothelin-1 has been demonstrated to contribute slightly to the maintenance of regional vascular tone. In gene-targeted mice, endothelin family peptides and their receptors have been shown to play an important role in the embryonic development of neural crest-derived tissues. In addition to its potent vasoconstrictor action, endothelin-1 has direct mitogenic actions on cardiovascular tissues, as well as co-mitogenic actions with a wide variety of growth factors and vasoactive substances. Endothelin-1 also promotes the synthesis and secretion of various substances including extracellular constituents. These effects of endogenous endothelin-1 would appear to be naturally concerned with the development and/or aggravation of chronic cardiovascular diseases, e.g. hypertension, pulmonary hypertension, vascular remodeling (restenosis, atherosclerosis), renal failure, and heart failure. A great many non-peptide and orally active endothelin receptor antagonists have been developed, and shown to exert excellent therapeutic effects in animal models as well as human patients with these diseases. PMID- 11725951 TI - Identification of arabidopsis proteins that interact with the cauliflower mosaic virus (CaMV) movement protein. AB - Gene I of cauliflower mosaic virus (CaMV) encodes a protein that is required for virus movement. The CaMV movement protein (MP) was used in a yeast 2-hybrid system to screen an Arabidopsis cDNA library for cDNAs encoding MP-interacting (MPI) proteins. Three different clones were found encoding proteins (MPI1, -2 and -7) that interact with the N-terminal third of the CaMV MP. The interaction in the 2-hybrid system between MPI7 and CaMV MP mutants correlated with the infectivity of the mutants. A non-infectious MP mutant, ER2A, with two amino acid changes in the N-terminal third of the MP failed to interact with MPI7, while an infectious second-site mutant, that differed from ER2A by only a single amino acid change, interacted in the 2-hybrid system. MPI7 is encoded by a member of a large, but diverse gene family in Arabidopsis. MPI7 is related in sequence, size and hydropathy profile to mammalian proteins (such as rat PRA1) described as a rab acceptor. The gene encoding MPI7 is expressed widely is Arabidopsis plants. and in transgenic plants the MPI7:GFP fusion protein is localized in the cytoplasm, concentrated in punctate spots. In protoplasts transfected with CFP:MP and MPI7:YFP, CFP:MP colocalized to some of the sites where MPI7:YFP is expressed. At these sites, fluorescence resonance energy transfer (FRET) between fluorophores was observed indicating an interaction in planta between the CaMV MP and MPI7. PMID- 11725954 TI - Tissue distribution of a major mevalonate pyrophosphate decarboxylase in rats. AB - The 45- and 35-kDa subunits of mevalonate pyrophosphate decarboxylase (MPD) have been purified from rat liver. In this study, we examined the relationship between 45- and 35-kDa MPD and the tissue distribution of a major MPD in rat liver. When the crude extract of rat liver fed on normal chow was subjected to immunoblot analysis using anti-rat 45-kDa MPD antibody, only the 45-kDa band was detected. In a pulse-chase experiment using anti-rat 45-kDa MPD antibody, there was no precursor-product relationship between the 45- and the 35-kDa MPD. In immunoprecipitation, more than 85% of MPD activity in the rat liver was depleted from the crude extract with an excess of the above antibody. When 45-kDa MPD contents in tissues were analyzed by immunoblotting, a single protein band with an apparent molecular weight of 45 kDa was detected in all tissues. The specific protein content of 45-kDa MPD in liver was markedly higher than in other tissues. The activity/amount ratio varied among brain, liver, and testis, being significantly highest in the liver. From these data, it is suggested that 45-kDa MPD serves as a major enzyme involved in cholesterol biosynthesis in rat liver and that a tissue-specific regulator or isozyme of 45-kDa MPD is present in rat liver. PMID- 11725952 TI - Expression and evolution of functionally distinct haemoglobin genes in plants. AB - Haemoglobin genes have been found in a number of plant species, but the number of genes known has been too small to allow effective evolutionary inferences. We present nine new non-symbiotic haemoglobin sequences from a range of plants, including class 1 haemoglobins from cotton, Citrus and tomato, class 2 haemoglobins from cotton, tomato, sugar beet and canola and two haemoglobins from the non-vascular plants, Marchantia polymorpha (a liverwort) and Physcomitrella patens (a moss). Our molecular phylogenetic analysis of all currently known non symbiotic haemoglobin genes and a selection of symbiotic haemoglobins have confirmed the existence of two distinct classes of haemoglobin genes in the dicots. It is likely that all dicots have both class 1 and class 2 non-symbiotic haemoglobin genes whereas in monocots we have detected only class 1 genes. The symbiotic haemoglobins from legumes and Casuarina are related to the class 2 non symbiotic haemoglobins, whilst the symbiotic haemoglobin from Parasponia groups with the class 1 non-symbiotic genes. Probably, there have been two independent recruitments of symbiotic haemoglobins. Although the functions of the two non symbiotic haemoglobins remain unknown, their patterns of expression within plants suggest different functions. We examined the expression in transgenic plants of the two non-symbiotic haemoglobins from Arabidopsis using promoter fusions to a GUS reporter gene. The Arabidopsis GLB1 and GLB2 genes are likely to be functionally distinct. The class 2 haemoglobin gene (GLB2) is expressed in the roots, leaves and inflorescence and can be induced in young plants by cytokinin treatment in contrast to the class 1 gene (GLB1) which is active in germinating seedlings and can be induced by hypoxia and increased sucrose supply, but not by cytokinin treatment. PMID- 11725955 TI - Mevalonate pyrophosphate decarboxylase is predominantly located in the cytosol of rat hepatocytes. AB - Mevalonate pyrophosphate decarboxylase (MPD) is found in the 100000 x g supernatant fraction of cells or tissues and is considered to be a cytosolic protein. Recently, other groups reported that MPD is mostly located in the peroxisomes. In this study, we used two different methods to determine whether MPD is predominantly located in the peroxisomes or the cytosol of rat hepatocytes. 1) In permeabilized rat hepatocytes or normal rat kidney cells treated with digitonin, which lack cytosolic enzyme, MPD was mainly present in the medium. 2) Double immunofluorescent labeling of cells with both anti-MPD antibody and anti-hexokinase antibody yielded an immunofluorescent pattern for both enzymes typical of the cytosolic protein. These results indicate that MPD is predominantly located in the cytosol of rat hepatocytes. PMID- 11725956 TI - Alterations of phospholipase C isozymes in rat cerebral cortex through hyperoxia. AB - The effect of hyperoxia on the level of three phospholipase C (PLC) isozymes (beta1, gamma1, delta1) was assessed in the rat cerebral cortex. When the rats were exposed to 100% oxygen for 60 h, there was a significant reduction in the catalytic activity of low molecular weight phosphotyrosine phosphatase, which was susceptible to activity loss under oxidative stress. The result suggests that oxidative stress is induced in the rat cerebral cortex through hyperoxia. The protein levels of PLC-beta1 and -delta1 were significantly increased in the cerebral cortex where oxidative stress had been induced, although that of PLC gamma1 was not altered. There was no significant difference in the total PLC activity of the cerebral cortex between hyperoxia and control rats. Using gel filtration chromatography, it was revealed that the PLC-beta1 activity in the cerebral cortex of the hyperoxia rats was higher than that in the control rats, but the PLC-delta1 activity in the former did not differ from that in the latter, despite an increase in the PLC-delta1 protein level. These findings suggest that the PLC-beta1 and -delta1 protein levels of brain tissues are increased by oxidative stress, and that the increased PLC-delta1 molecule is less active. PMID- 11725957 TI - Effect of calpain on hereditary cataractous rat, ICR/f. AB - The crystallins in the lenses of ICR/f mutation rat, a known hereditary cataract model, were analyzed during cataractogenesis. Opacification of the mutant lenses was found to be accompanied by changes in crystallin structure and composition, including several deletions of the N-terminals of beta-crystallins and low molecular weight alpha- crystallins. Because similar deletions were observed when the soluble fraction of normal lens protein was incubated with calpain, we considered that calpain could be related to the deletions in mutant lenses. Although measurement of the content of calpain protein by the ELISA method revealed no significant difference between mutant and normal lenses, it was found that the concentrations of Ca2+ and K+ were different between the two lenses and that calpain activity was dependent on both ion concentrations. Endogenous m calpain in the soluble fraction from normal lenses was activated by addition of 1 mm calcium chloride in the presence of 50 mm KCl (the same concentration as in mutant lenses), and insoluble protein was found in the fraction 1 d after calpain activation. On the other hand, the presence of 120 mm KCl (the concentration in normal lenses) inhibited calpain activity and prevented this insolubilization. These results suggest that calpain in mutant lenses is involved in the proteolysis of crystallins and the progression of cataract formation. PMID- 11725958 TI - Antioxidative properties of pig vesical mucosa: a comparison with gastric and intestinal mucosa. AB - The antioxidative properties of pig urinary bladder mucosa were compared with those of gastric and intestinal mucosa using nitroxide radicals. Electron paramagnetic resonance (EPR) method was used to monitor the metabolic processes of nitroxides in mucosae. The reduction of nitroxides was measured on intact luminal surfaces of gastric, intestinal, and urinary bladder mucosa, as well as in homogenates of mucosa surface layer. Furthermore, N-ethylmaleimide and ascorbate oxidase have been used to characterize the reducing agents in urinary bladder mucosa homogenates. The nitroxide concentration decrease on intact mucosa of the urinary bladder was significantly different from those of the gastric and the intestinal mucosa. The concentration decrease was the largest for intestinal mucosa and the smallest for bladder mucosa. On the other hand, homogenates exhibit the largest nitroxide reduction rates for the bladder mucosa and the smallest for the gastric mucosa. In the bladder surface layer homogenates ascorbate and thiol-containing reducing agents were found and their coupled action in the nitroxide reduction process was established. The mucosa of urinary bladder is protected against nitroxide free radicals by a relatively low permeability and very active endogenous reducing agents. The gastric and intestinal mucosa are more permeable and/or have greater antioxidant activity on their surface. The reduction of nitroxides in the urinary bladder mucosa occurs via the ascorbate-thiol coupled reducing system. PMID- 11725959 TI - Expression profiling of sulfotransferases in human cell lines derived from extra hepatic tissues. AB - To explore the physiological roles of sulfotransferases (SULTs) in extra-hepatic tissues, we examined the expression of eight SULT genes by reverse transcription (RT)-PCR in human cell lines that were established from various tissues. Expression levels of SULTs were low in neural cell lines such as NB-1 and GI-1, and high in epithelial cell lines, such as Caco-2 and BeWo. SULT1C2 expression was abundant in all cell types, whereas that of SULT1E1, SULTIBI or SULT2B1 was restricted to a specific cell type. SULT1C1, which can catalyze the sulfation of N-hydroxy-2-acetylaminofluorene, was expressed in Caco-2, BeWo and KB562. Induction of differentiation did not generally affect SULT expression, although that of SULT1C2 was reduced after differentiation of the neuroblastoma cell line, NB-1, was induced. The profile of SULT expression in the culture cells obtained here gives clues to understanding the physiological roles of SULT enzymes in extra-hepatic tissues or organs. PMID- 11725960 TI - Identification of human drug-metabolizing enzymes involved in the metabolism of SNI-2011. AB - In vitro studies were conducted to identify human drug-metabolizing enzymes involved in the metabolism of SNI-2011 ((+/-)-cis-2-methylspiro [1,3-oxathiolane 5,3'-quinuclidine] monohydrochloride hemihydrate, cevimeline hydrochloride hydrate). When 14C-SNI-2011 was incubated with human liver microsomes, SNI-2011 trans-sulfoxide and cis-sulfoxide were detected as major metabolites. These oxidations required NADPH, and were markedly inhibited by SKF-525A, indicating that cytochrome P450 (CYP) was involved. In a chemical inhibition study, metabolism of SNI-2011 in liver microsomes was inhibited (35-65%) by CYP3A4 inhibitors (ketoconazole and troleandomycin) and CYP2D6 inhibitors (quinidine and chlorpromazine). Furthermore, using microsomes containing cDNA-expressed CYPs, it was found that high rates of sulfoxidation activities were observed with CYP2D6 and CYP3A4. On the other hand, when 14C-SNI-2011 was incubated with human kidney microsomes, SNI-2011 N-oxide was identified as a major metabolite. This N oxidation required NADPH, and was completely inhibited by thiourea, indicating that flavin-containing monooxygenase (FMO) was involved. In addition, microsomes containing cDNA-expressed FMO1, a major isoform in human kidney, mainly catalyzed N-oxidation of SNI-2011, but microsomes containing FMO3, a major isoform in adult human liver, did not. These results suggest that SNI-2011 is mainly catalyzed to sulfoxides and N-oxide by CYP2D6/3A4 in liver and FMOI in kidney, respectively. PMID- 11725961 TI - Induction of histamine release from rat peritoneal mast cells by histatins. AB - Human salivary histatins (Hsts), which belong to a salivary polypeptide family, have potent antifungal activity against Candida albicans and Cryptococcus neoformans, and are expected to be useful as therapeutic reagents against Candida species. However, little is known about the effect of Hsts on host immune systems. Thus we conducted a series of in vitro experiments with rat mast cells to determine whether histatin 5 (Hst 5) or histatin 8 (Hst 8) has a histamine releasing effect on mast cells. Both Hst 5 and Hst 8 induced histamine release from rat peritoneal mast cells in a dose-dependent manner (10(-9) to 10(-5) M). Hst 5 had a stronger releasing effect than Hst 8. The histamine release induced by Hst 5 (10(-6) M) was increased by the presence of 0.5 mM Ca2+, but decreased by 2mM Ca2+. Alternatively, the histamine release induced by Hst 8 (10(-6) M) was inhibited by the presence of Ca2+ (0.5 to 2 mM). These results suggest that Hsts have limited usefulness as therapeutic agents due to induction of histamine release from mast cells. PMID- 11725962 TI - The protective effects of PMC against chronic carbon tetrachloride-induced hepatotoxicity in vivo. AB - In this study, PMC (2,2,5,7,8-pentamethyl-6-hydroxychromane), a derivative of alpha-tocopherol, dose-dependently (1-10 mg/kg) ameliorated the increase in plasma aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) levels caused by chronic repeated carbon tetrachloride (CCl4) intoxication in mice. Moreover, PMC significantly improved the CCl4-induced increase of hepatic glutathione peroxidase, reductase, and superoxide dismutase activities. PMC also restored the decrement in the glutathione content of hepatic tissues in CCl4 intoxicated mice. Furthermore, it also dose-dependently inhibited the formation of lipid peroxidative products during carbon tetrachloride treatment. Histopathological changes of hepatic lesions induced by carbon tetrachloride were significantly improved by treatment with PMC in a dose-dependent manner. These results suggest that PMC exerts effective protection in chronic chemical-induced hepatic injury in vivo. PMID- 11725963 TI - Gastric-mucous membrane protection activity of coptisine derivatives. AB - Coptisine and 8-oxocoptisine were isolated as principles of the gastric-mucous membrane protection from Coptidis rhizoma. The two compounds showed stronger activity than cimetidine and sucralfate. We prepared several derivatives having a partial structure of coptisine from commercially available starting materials. The compounds obtained were tested for gastric-mucous membrane protective activity and a correlation between activity and structure was studied. Our results suggest that the partial charge of the catechol skeleton is related to activity. PMID- 11725964 TI - Cancer chemopreventive activity of odorine and odorinol from Aglaia odorata. AB - In the course of our continuing search for novel cancer chemo-preventive agents from natural sources, we have carried out a primary screening in vitro assay of the compounds isolated from Aglaia odorata. Consequently, aminopyrrolidine diamides, odorine and odorinol, were obtained as active constituents. These compounds exhibited potent anti-carcinogenic effects in a two-stage carcinogenesis test of mouse skin induced by 7,12-dimethylbenz[a]anthracene (DMBA) as an initiator and 12-O-tetradecanoylphorbol-13-acetate (TPA) as a promoter. Further, both compounds showed remarkable inhibitory effects in two stage mouse skin carcinogenesis models induced by nitric oxide (NO) donors such as (+/-)-(E)-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hexenamide (NOR-1) or peroxynitrite as an initiator and TPA as a promoter. From these results, it was concluded that odorine and odorinol inhibited both the initiation and promotion stages of two-stage skin carcinogenesis. PMID- 11725965 TI - Cytotoxic activities and structure-cytotoxic relationships of steroidal saponins. AB - We have systematically examined the cytotoxic activities of the steroidal saponins mainly isolated from the Liliaceae plants against HL-60 human promyelocytic leukemia cells and found several structure-activity relationships. Some steroidal saponins evaluated in the assay system showed considerable cytotoxic activities, which were almost as potent as that of etoposide used as a positive control. The activities were found to be sensitive to the monosaccharides constituting the sugar moieties and their sequences, as well as to the structures of the aglycons. PMID- 11725966 TI - Pharmacokinetic and pharmacodynamic analysis of the antihypercalcemic effect of incadronate disodium in rats. AB - Incadronate concentrates into the bone as a target organ after intravenous administration of incadronate disodium. Mature osteoclasts has take up incadronate from the bone surface and convert it from an active to an inactive form. As a result, incadronate decreases the plasma calcium concentration by suppressing bone resorption. In this study, the pharmacokinetic and pharmacodynamic (PK/PD) analysis model for ascertaining the antihypercalcemic effects of incadronate disodium was developed in rats. Data on both the concentration of incadronate in bone and that of free calcium in blood after intravenous administration from our previous study were used for analysis. To estimate the concentration in the surface layer of bone, data on the concentration of incadronate in bone after single intravenous administration were analyzed based on the PK model considering three-compartments. The estimated concentrations in the surface layer in bone were applied to the PD model as an input function. The PD model was developed to analyze the changes in the plasma calcium concentration after a single intravenous administration considering an irreversible inhibition of osteoclast activity. The obtained fitted curves were in good agreement with the observed data. The model could explain the long duration of the antihypercalcemic effect of incadronate disodium and should be useful for planning rational dose regimens for effective antihypercalcemic therapy. PMID- 11725967 TI - Relative hypoglycemic effect of insulin suppositories in diabetic beagle dogs: optimization of various concentrations of sodium salicylate and polyoxyethylene-9 lauryl ether. AB - The effect of insulin suppositories containing different amounts and concentrations of sodium salicylate (50, 100 mg) and polyoxyethylene-9-lauryl ether (POELE 1, 3, 4%), respectively, on the plasma glucose concentration of diabetic beagle dogs was investigated after rectal administration. Comparison of the effects of these formulations was made with that produced after subcutaneous insulin injections. Insulin suppositories containing sodium salicylate (50 mg) produced a maximum reduction of plasma glucose concentration (Cmax) of 55 +/- 11%, an area under the curve (AUC) of 252 +/- 59% reduction h; and a relative hypoglycemia (RH) of 49 +/- 12% relative to subcutaneous injection of insulin (4 U/kg). Increasing sodium salicylate to 100 mg/suppository did not improve the hypoglycemic effect of insulin suppositories further. Investigation of the influence of insulin suppositories containing different concentrations of the nonionic surfactant POELE (1, 3, 4%) showed that; the suppositories containing the lowest concentration (1%) produced the highest hypoglycemic effect with a Cmax of 68%, AUC of 332 +/- 67% reduction h, and RH of 55 +/- 11%. Incorporation of sodium salicylate 50mg in insulin suppositories containing 1% POELE did not improve further the effects found with these suppositories. In conclusion, a relative hypoglycemic effect of about 50-55% can be achieved using insulin suppositories containing Witepsol W35 as a base, insulin (5 U/kg), and sodium salicylate (50 mg) or POELE (1%) as rectal absorption enhancers. PMID- 11725968 TI - Introduction of recirculatory analysis into portal and systemic concentration difference method. AB - Recirculatory analysis was introduced into the portal and systemic concentration difference method with double dosing (PS-DD method), which is an evaluation system for the local intestinal and hepatic first-pass effect. 5-Fluorouracil (5 FU) and cephalexin (CEX) were selected as model drugs. A new recirculatory system was constructed to predict the time courses of a drug concentration in the systemic and portal bloods. Bioavailability (F), local absorption ratio (Fa), hepatic recovery ratio (FH), and local mean absorption time (ta) estimated by recirculatory analysis were close to those calculated by moment analysis with numerical integration. Using recirculatory analysis, the sampling period was considerably shortened and the sampling number was also reduced, which demonstrates that recirculatory analysis is useful in PS-DD method. PMID- 11725969 TI - Application of the PKCYP-test to predict the amount of in vivo CYP2C11 using tolbutamide as a probe. AB - Previous reports have shown that the determination of drug metabolism capacity can be made by the pharmacokinetic estimation of the quantity of cytochrome P450 (CYP) in vivo (PKCYP-test), in which an apparent liver-to-blood free concentration gradient in vivo (qg) is introduced, which is useful for evaluating fluctuations of CYPIA2 in rats. The aim of the present study was to examine the application of the PKCYP-test to evaluate the quantity of in vivo CYP2C11 by using tolbutamide as a probe, to confirm its validity using a physiologically based pharmacokinetic rat model. Rats treated with carbon tetrachloride (CCl4 treated rats) were used as a model for low levels of CYP2C11 in the liver. In CCl4-treated rats, the total body clearance (CLtot) of tolbutamide and the amount of CYP2C11 fell to about a quarter and a third of that in control rats, respectively. The time-course of tolbutamide concentrations in serum in control rats could be simulated by a physiologically-based pharmacokinetic model. In CCl4 treated rats, take into consideration the qg value of control rats, the level of CYP2C11 was accurately predicted by the PKCYP-test, and the time-course of tolbutamide concentrations in serum could be predicted by the same physiologically-based pharmacokinetic model. In conclusion, we have shown that the PKCYP-test can be used to predict levels of CYP2C11. It was also demonstrated that the qg and amount of CYP are useful parameters in the PKCYP-test by constructing a physiologically-based pharmacokinetic model which was applied to the PKCYP-test. PMID- 11725970 TI - De novo regeneration of Scrophularia yoshimurae Yamazaki (Scrophulariaceae) and quantitative analysis of harpagoside, an iridoid glucoside, formed in aerial and underground parts of in vitro propagated and wild plants by HPLC. AB - A protocol for de novo regeneration and rapid micropropagation of Scrophularia yoshimurae (Scrophulariaceae) has been developed. Multiple shoot development was achieved by culturing the shoot-tip, leaf-base, stem-node and stem-internode explants on Murashige and Skoog (MS) medium supplemented with 4.44 microM N6 benzyladenine (BA) and 1.07 microM alpha-naphthaleneacetic acid (NAA). Stem-node and shoot-tip explants showed the highest response (100%) followed by stem internode (74.4%) and leaf-base (7.7%) explants. The shoots were multiplied by subculturing on the same medium used for shoot induction. Shoots were rooted on growth regulator-free MS basal medium and the plantlets were transplanted to soil and acclimatized in the growth chamber. The content of harpagoside, a quantitatively predominant iridoid glycoside, in different plant material was determined by high performance liquid chromatography (HPLC). The analysis revealed that the content of harpagoside in the aerial and underground parts of S. yoshimurae was significantly higher than the marketed crude drug (underground parts of Scrophularia ningpoensis). PMID- 11725971 TI - Characterization of a Ca2+-dependent protein kinase from rice root: differential response to cold and regulation by abscisic acid. AB - The Ca2+-dependent protein kinases (CDPKs) and abscisic acid (ABA) are known to be involved in low-temperature stress response. The focus of this study was to characterize the 45 kDa protein kinase identified in the crude extract of rice (Oryza sativa L.) seedling roots in response to cold (5 degrees C) stress. The activity of the 45 kDa protein kinase decreased at low temperature as evident by an in-gel kinase assay using histone III-S as a substrate. Also, the Ca2+ dependent activity of this protein kinase was suppressed by cold in the membrane fractions of the root. A general protein kinase inhibitor and Ca2+ chelator inhibited the activity of the 45 kDa protein kinase, suggesting that it was a plant CDPK. The 45 kDa CDPK identified was found to be independent of photosynthetic tissues such as the leaf and leaf sheath of rice seedlings, supporting a direct sensing mechanism in the roots of rice seedlings to cold stress. The suppressed activity of the 45 kDa CDPK was reverted by supplementing with 5 microM ABA under cold stress. The 45 kDa CDPK activity was stronger in the cold-tolerant variety of the 4 types tested than it was in the cold-sensitive one. These results suggest the involvement of endogenous ABA in regulating the activity of the 45 kDa CDPK in response to cold stress. PMID- 11725972 TI - Effect of tenascin-X together with vascular endothelial growth factor A on cell proliferation in cultured embryonic hearts. AB - Tenascin-X (TNX) is a large glycoprotein that appears in extracellular matrices. Previously, we demonstrated that TNX binds to vascular endothelial growth factors A and B (VEGF-A and -B) and that VEGF-B in combination with TNX induces DNA synthesis in endothelial cells via increased signals mediated by the VEGFR-1 receptor. In this study, we investigated the effect of TNX with VEGF-A on the cell proliferation in embryonic mouse heart explants from either wild-type (TNX+/+) or TNX-deficient (TNX-/-) mice. The addition of VEGF-A to the explants from TNX+/+ mice increased cell proliferation by 1.5 fold compared with that in TNX-/- mice, indicating that TNX with VEGF family member plays an important role in the control of endothelial cell proliferation in vivo. PMID- 11725973 TI - Initial intraorgan formation of mercapturic acid. AB - The disposition of S-benzyl-glutathione (BSG) in male Wistar rats was evaluated by the HPLC method to examine whether the kidney and liver contributed independently to the biosynthesis of S-benzyl-N-acetylcysteine (BNAc), a mercapturic acid (Chart 1). After intravenous injection, BSG was rapidly transported in both the kidney and the liver at a ratio of about 7:3. Simultaneously, a large amount of BNAc was found in both the kidney and the liver. In the kidney, S-benzyl-cysteine (BCys) reached a maximum concentration (Cmax) at 2 min after BSG injection, whereas BNAc reached Cmax within 3 to 5 min. The generation of BNAc was also observed in the liver. While renal BNAc reached Cmax within 3 to 5 min, hepatic BNAc reached Cmax around 5 min after BSG injection. Moreover, the elimination half-life of the BNAc after intravenous injection of the BSG was equivalent to that observed after intravenous injection of the BNAc itself. These results demonstrate that the kidney contributes to the initial intraorgan generation of BNAc and that this mercapturic acid is also synthesized in the liver and preferentially excreted into urine. PMID- 11725975 TI - Activation of mu-opioid pathway is associated with the canceling effect of footshock stimulus on the restraint stress-induced inhibition of small intestinal motility in rats. AB - We previously reported that small intestinal motility was significantly inhibited by restraint stress, but not by footshock stress. In the present study, we found that plasma beta-endorphin levels were more significantly elevated by footshock stress than restraint stress, and that preloading of footshock stimulus canceled the inhibition of small intestinal motility by restraint stress. Pretreatment with the mu-opioid receptor antagonist naltrexone significantly attenuated this canceling effect of footshock stimulus. These results suggest that footshock stimulus may cancel the inhibition of small intestinal motility by restraint stress via activation of mu-opioid receptors. PMID- 11725974 TI - Plasma, intestine and tumor levels of 5-fluorouracil in mice bearing L1210 ascites tumor following oral administration of 5-fluorouracil, UFT (mixed compound of tegafur and uracil), carmofur and 5'-deoxy-5-fluorouridine. AB - Several 5-fluorouracil (5-FU) derivatives, 1-hexylcarbamoyl-5-fluorouracil (HCFU), 5'-deoxy-5-fluorouridine (5'-DFUR) and UFT (mixed compound of tegafur and uracil), have been developed and clinically widely used. However, comparative pharmacokinetic studies of the parent compound and other fluorinated drivatives have not been precisely reported. The dosage of the oral clinical use for human cancer of 5-FU, HCFU, 5'-DFUR and UFT as tegafur (FT) is 200-300mg/d, 600mg/d, 800-1,200mg/d and 300-600mg/d respectively. These amounts of the drugs are almost equimolar. Previously, we reported the effect of oral equimolar administration of each four drugs on thymidilate synthase activity, deoxyribonucleotide metabolism and cell cycle progression in L1210 ascites tumor. (1,2) In this study, we examined the antitumor effect and 5-FU concentration in the plasma, intestine and tumor after oral equimolar administrations of each drug using BDF1 mice bearing L1210 ascites tumor. In our study, UFT showed the best life prolongation among these four drugs. The intestine 5-FU level was highest by treatment with 5-FU during the initial 4 h. The plasma 5-FU level was highest by treatment with HCFU for 4 h. But the tumor 5-FU level was highest by treatment with UFT over the 24 h. In spite of the high plasma 5-FU concentration after the treatment with HCFU, the 5-FU concentration in the tumor was below the detectable level until 24 h. These findings suggested that the highest specific accumulation of 5-FU in tumor cells may explain the best therapeutic results of UFT. PMID- 11725976 TI - Trifluoromethyl ketone-based inhibitors of apoptosis in cerebellar granule neurons. AB - A variety of aromatic trifluoromethyl ketone derivatives has been studied as inhibitors of apoptosis in cerebellar granule neurons (CGNs). Among them, alpha trifluoromethyl diketone (2) and benzyl trifluoromethyl ketone (11) were found to be apoptosis inhibitors which can prevent a neurodegenerative disease. Compounds 2 and 11 showed neuroprotection effect on low K+-induced apoptosis in CGNs. Furthermore, these compounds effectively suppressed DNA fragmentation accompanied with apoptosis. The neuroprotection mode of 2 and 11 was not related to inhibition of caspase-3. PMID- 11725977 TI - Antiproliferative constituents in the plants 7. Leaves of Clerodendron bungei and leaves and bark of C. trichotomum. AB - The constituents of the leaves of Clerodendron bungei STEUD. (Verbenaceae) and leaves and bark of C. trichotomum THUNB. were investigated guided by the antiproliferative activity against three tumor cell lines (MK-1: human gastric adenocarcinoma, HeLa: human uterus carcinoma, and B16F10: murine melanoma). Two phenylethanoid glycoside caffeic acid esters, acteoside and isoacteoside, were isolated as the constituents which selectively inhibit the growth of B16F10 cells. The antiproliferative activities against B16F10 cells of acteoside (GI50: 8 microM), isoacteoside (8 microM) and their methanolysis products, methyl caffeate (26 microM), 3,4-dihydroxyphenethyl alcohol (8 microM), 3,4 dihydroxyphenethyl glucoside (10 microM), desrhamnosyl acteoside (6 microM), and desrhamnosyl isoacteoside (6 microm) suggested that the 3,4-dihydroxyphenethyl alcohol group might be more responsible for the activities of acteoside and isoacteoside than the caffeoyl group. The activities of chlorogenic acid, 3,4 dihydroxyphenylacetic acid, 3-(3,4-dihydroxyphenyl) alanine, 3,4-dihydroxy phenethylamine hydrochloride, ferulic acid, sinapic acid, and five dihydroxybenzoic acids were also determined and compared with those of the above compounds. PMID- 11725978 TI - Therapeutic use of Bausch & Lomb PureVision contact lenses. AB - PURPOSE: To evaluate the Bausch & Lomb PureVision contact lens as a continuous wear contact lens for therapeutic indications in a prospective open-ended non randomized clinical trial. METHODS: Patients with a variety of corneal and ocular surface disease conditions presenting at the Singapore National Eye Centre who required therapeutic continuous contact lens wear were enrolled. Therapeutic indications included pain relief, corneal protection, and enhancement of corneal wound healing. Success or failure of specific treatment indications was assessed in all cases, with evaluation of lens performance and fit characteristics, and the presence of ocular complications or lens-related complications was noted. RESULTS: There were 54 patients (54 eyes), and the mean duration of continuous contact lens wear was 1.1 months. Conditions treated included post-surgical indications (n = 36) (post-keratoplasty or ocular surface transplantation, post LASIK or PRK surgery) bullous keratopathy (n = 7), chemical burns (n = 3), epithelial abrasions or recurrent corneal erosion syndromes (n = 3), corneal perforations (n = 3), neurotrophic ulcer (n = 1), and corneal laceration (n = 1). For the indication of corneal healing (40 eyes), improved healing was noted in 38 eyes (96%), with full healing occurring in 33 eyes (83%). For pain relief (28 eyes), 27 patients (96%) had considerable or complete pain relief, and the remaining patient reported partial pain relief. For corneal protection (21 eyes), lens wear was fully protective in all cases. The lens performance and fitting characteristics surpassed any previous therapeutic lenses used by the investigators. Complications related to contact lens wear were limited to one case of a culture-negative corneal infiltrate requiring cessation of therapeutic lens wear and one case of a loosely fit lens. CONCLUSIONS: Our results show that the PureVision contact lens exhibits good safety and efficacy when utilized as a continuous wear therapeutic lens. With the theoretical advantage of increased oxygen transmissibility reducing the risk of hypoxia-related complications, this new lens may be one step closer to the ideal therapeutic contact lens. The PureVision contact lens is now our primary bandage lens of choice in our corneal, external disease, and refractive surgery services. PMID- 11725979 TI - Diffusion limited evaporation rates in hydrogel contact lenses. AB - PURPOSE: To systematically measure evaporative water loss for a range of commercial hydrogel contact lens types, correlate the results with measured water self-diffusion coefficients and determine the best way to present and quantify the in vitro evaporation data. METHODS: Evaporative water loss was measured gravimetrically for the four commercial contact lens types: Acuvue (powers: -0.50 D, -0.75 D), Newvues (powers: -0.50 D, -0.75 D, -1.00 D), CIBAsoft (powers: -0.50 D, -0.75 D, -1.00 D) and SeeQuence 2 (power: -0.75 D), with water contents of 60%, 55%, 38%, and 38%, respectively. Water proton self-diffusion coefficients were measured for these materials using standard nuclear magnetic resonance (NMR) pulsed field gradient techniques. RESULTS: The initial rate of evaporation was linear for 5-10 minutes then decreased in all cases. No significant differences were observed in this initial rate between the lenses studied. However, the evaporative rate decreased more slowly with time for the higher water content lenses. The diffusion coefficients were found to be highly dependent on the equilibrium water content, regardless of material composition. CONCLUSIONS: The most effective way to present/quantify this type of evaporative data is to plot the time-course of the mass evaporation rate. We have found that the decrease in evaporative rate with time is a diffusion-limited effect whereby lenses with higher water content can maintain a higher rate of diffusion of water (to the evaporative surface) due to the presence of higher proportions of mobile water molecules. The NMR diffusion data corroborate these findings. PMID- 11725980 TI - Conditioning of hydrogel lenses by a multipurpose solution containing an ocular lubricant. AB - PURPOSE: The addition of the ocular lubricant hydroxypropyl methylcellulose (HPMC) to a multipurpose contact lens solution conditions the hydrogel lens surface. This investigation reports the clinical benefits to contact lens wearers and the improved physical properties of the solution with HPMC added. METHODS: One-hundred forty-seven subjects wearing a variety of hydrogel lenses used a multipurpose solution with and without addition of HPMC and were assessed for product comfort and preference. In the laboratory, conditioning by HPMC was demonstrated in fluid coating, dynamic contact angle, uptake of fluid on hydrogel lenses, and release of HPMC from lenses after soaking. RESULTS: Lens wearers reported the multipurpose solution with HPMC more comfortable, including measures relating to maintaining lens moisture over time. The multipurpose solution with HPMC produced a thicker and longer-lasting layer of fluid on hydrogel lenses and other plastics than other multipurpose solutions. HPMC was found to adsorb to both group 1 and 4 lens materials and release gradually, with detectable amounts releasing from the lens beyond 12 hours. CONCLUSIONS: The conditioning properties of the multipurpose solution with HPMC produce improved wetting of lenses and enhanced lens wearing comfort. Binding of HPMC to the lens surface and subsequent time-release is the probable mechanism for these benefits. PMID- 11725981 TI - Rigid gas-permeable contact lenses are a safe and effective means of treating refractive abnormalities in the pediatric population. AB - PURPOSE: This study examined the safety, efficacy, and tolerance of rigid gas permeable (RGP) contact lenses in a pediatric population. The study also considered the economic impact of RGP contact lens wear in children. METHODS: Retrospectively, we reviewed 12 consecutive charts of children with multiple diagnoses of trauma (n = 5), high myopia (n = 1), surgical aphakia (n = 5), corneal scar (n = 4), and microophthalmia (n = 2) who were considered for RGP contact lenses. RESULTS: Fifteen eyes of 12 patients were originally fit with RGP contact lenses. Ten of 12 (83.3%) children were still wearing their RGP contact lenses after a mean follow-up of 17.8 months. Two of 12 patients (16.7%) stopped RGP use. Two of 10 (20%) patients wore their RGP contact lenses for extended wear. Eight of 10 (80%) patients were successful in daily insertion and removal of RGP contact lenses. There were no adverse events secondary to contact lens use in any patients. An average of 2.18 (range: 0.97-4.55) replacement lenses were needed per patient per 6 month interval. CONCLUSIONS: RGP contact lenses are a safe, effective, and tolerable means of treating refractive abnormalities in the pediatric population. With proper education, families can be successful with daily RGP insertion and removal. RGP contact lenses have the advantage of excellent oxygen transmissibility, low bacterial and protein adherence, and the potential to correct irregular astigmatism. Disadvantages include frequent lens replacement. PMID- 11725982 TI - Use of SoftPerm contact lenses when rigid gas permeable lenses fail. AB - PURPOSE: We evaluated the performance of the SoftPerm contact lens (Wesley Jessen) in patients with irregular astigmatism, usually due to keratoconus or after penetrating keratoplasty (PK), who were unable to befitwith, or intolerant of, rigid gas permeable (RGP) contact lenses. METHODS: A retrospective study of patients fit with SoftPerm lenses in the Cornea Department at Wills Eye Hospital between March 1985 and March 2000 was performed. RESULTS: Thirty-five cases were reviewed, with follow-up available in 33 cases. Most of the eyes had irregular astigmatism secondary to keratoconus (22/35,62.9%) or PK (10/35,28.6%) and had failed a trial of RGP lenses. The mean logMAR visual acuity with SoftPerm lenses was 0.13+/-0.18 (range -0.12 to 0.6). In 25 cases in which comparison with glasses or RGP lenses was possible, SoftPerm lenses provided better visual acuity than glasses in 17/25 cases (68%) with a mean difference of -0.24 (P = 0.001, paired t-test); visual acuity with SoftPerm lenses was better than RGP visual acuity in 13/25 cases (52%), with a mean difference of -0.06 (P = 0.07, paired t test). Complications included broken lenses (16/33,48.5%), giant papillary conjunctivitis (GPC) (9/33, 27.3%), and peripheral corneal neovascularization (9/33, 27.3%). The GPC and peripheral corneal neovascularization were often delayed in presentation. The major subjective complaint was discomfort (13/33, 39.4%). At the last follow-up, the SoftPerm lens was still in use in 22/33 cases (66.7%). Discomfort was the most common reason for discontinuation. The mean duration of lens wear was 52.5+/-31.7 months, range 3 to 110 months. CONCLUSIONS: The SoftPerm lens can provide satisfactory visual correction in many cases of irregular astigmatism with RGP failure. However, problems such as frequent breakage, GPC, peripheral corneal neovascularization, and discomfort necessitate close follow-up. PMID- 11725983 TI - Chlamydial conjunctivitis in contact lens wearers: successful treatment with single dose azithromycin. AB - PURPOSE: To reveal clinical presentations of chlamydial conjunctivitis in contact lens wearers as well as to evaluate the clinical and microbiological efficacy of oral azithromycin in the treatment of this condition. METHODS: Twenty contact lens users with chlamydial conjunctivitis were included in this retrospective study. Chlamydial infection was diagnosed by isolation of Chlamydia trachomatis in cell culture of conjunctival scrapings. All patients were treated with a single 1 g oral dose of azithromycin. Follow-up clinical and microbiological examinations were performed 1 month after treatment. RESULTS: All patients suffered from some ocular symptoms such as itching, burning, tearing, and nonspecific irritation, but none had apparent conjunctival injection or any conjunctival discharge. The majority (90%) had bilateral complaints. Mild follicular reaction, limited to the lateral part of lower fornices, was present in 17 patients; the remaining patients had normal biomicroscopical findings. Four weeks following the single azithromycin dose, C. trachomatiswas eradicated in all patients and 17 (85%) were free of symptoms. CONCLUSION: Chlamydial infection should be considered more frequently in differential diagnosis of symptomatic contact lens wearers. Azithromycin is the most promising agent for the treatment of chlamydial conjunctivitis due to its excellent bacteriological efficacy and very convenient single dose administration. PMID- 11725984 TI - Disposable 1-day Acuvue contact lenses for the delivery of lomefloxacin to rabbits' eyes. AB - PURPOSE: To investigate the ability of a disposable soft contact lens (1-Day Acuvue) to deliver lomefloxacin, a fluoroquinolones antibiotic, into the cornea and aqueous humor of rabbits. METHODS: Experiments were conducted on adult albino rabbits. 1-Day Acuvue lenses were immersed for 1 hour in commercially-available lomefloxacin eye solution and then placed on the cornea. After 0.5, 2, 4,6, and 8 hours, the animals were killed and the lenses were removed and placed into a 20 mL saline bath. Corneal tissue and aqueous humor samples were also obtained at these times. The release of lomefloxacin from the lenses was calculated by measuring the amount of drug remaining in the lenses after removal from the rabbit eyes. The concentration of lomefloxacin in the cornea and anterior chamber following the wearing of lomefloxacin-loaded lenses was compared with the concentrations following frequent-drop therapy (one drop of lomefloxacin hourly for 8 hours). RESULTS: In vivo, lomefloxacin was released from 1-Day Acuvue lenses gradually over 8 hours. The cornea and aqueous humor levels in the eyes following the wear of lomefloxacin-loaded lenses were significantly higher than those achieved by frequent-drop therapy. The drug levels in the cornea and aqueous humor generally remained above the 90% minimal inhibitory concentration for 8 hours in the 1-Day Acuvue lens group. CONCLUSIONS: 1-Day Acuvue contact lenses soaked in 0.3% lomefloxacin can release sufficient amounts of lomefloxacin and in this study produced higher levels in both the cornea and aqueous humor than that achieved by frequent-drop therapy for up to 8 hours. We conclude that 1 Day Acuvue contact lens can be used as a drug delivery system for lomefloxacin. PMID- 11725985 TI - Studies on the uptake and release of fluoroquinolones by disposable contact lenses. AB - PURPOSE: To evaluate the ability of three types of disposable contact lenses to take up and release three kinds of fluoroquinolones in vitro. METHODS: We evaluated three FDA-approved disposable soft contact lenses (1-Day Acuvue, Medalist, and 14UV) for their ability to absorb and release three kinds of fluoroquinolones. Contact lenses were presoaked in fluoroquinolones for 1, 4, and 24 hours, and the uptake was determined by measuring the concentration of fluoroquinolones in the three types of disposable soft contact lenses by high pressure liquid chromatography (HPLC). After uptake, the lenses were placed in fresh saline baths, and release rates from the lenses were determined by measuring the concentration of fluoroquinolones in the saline baths by HPLC. RESULTS: The disposable soft contact lens with the highest uptake of fluoroquinolones was the 1-Day Acuvue. After presoaking, drug concentrations in the 1-Day Acuvue and Medalist were higher than their theoretical saturation concentrations (i.e., the concentration of the fluoroquinolone eye drops [0.3%]). The release rates from the 1-Day Acuvue and Medalist lenses were slower than for the 14 UV lens. CONCLUSIONS: These results indicate that among the lenses tested, the most practical drug delivery system is the 1-Day Acuvue disposable soft contact lens. PMID- 11725986 TI - Epitope variability of Bcl-2 immunolocalization in the human corneal epithelium. AB - PURPOSE: To further study the immunological localization of Bcl-2 protein in human corneal epithelium. METHODS: Three anti-human Bcl-2 antibodies, generated against amino acid residues (aa) 4-21 (polyclonal), 1-205 (monoclonal), and 41-54 (monoclonal), were used to localize Bcl-2 protein immunocytochemically in fresh eye bank donor human corneas. RESULTS: In the central corneal epithelium, two anti-Bcl-2 antibodies (aa 4-21 and aa 1-205) showed intense cytoplasmic staining of basal epithelial cells. These antibodies produced similar staining in the limbal epithelium, with gradually less intense staining of wing and superficial cells. By contrast, as previously reported, a monoclonal antibody to aa 41-54 stained nuclei of all epithelial cell layers with the exception of some surface corneal epithelial cells; this antibody also demonstrated very bright anti-Bcl-2 staining of Langerhans cells localized in the peripheral corneal epithelium. CONCLUSION: In our previous study, Bcl-2 protein was immunocytochemically localized to the nuclear compartment of all corneal epithelial cell layers with the use of antibodies specific for the regulatory flexible loop domain of Bcl-2. However, Bcl-2 can also be uniquely localized to the cytoplasm of the corneal epithelium with the use of antibodies generated against aa 4-21 and aa 1-205. Taken together, these results using epitope specific antibodies indicate that different epitopes on the Bcl-2 protein are available for antibody binding within different cells and cellular compartments, suggesting that proliferation and differentiation may lead to changes in the Bcl-2 structure and conformation within different compartments of the epithelial cells themselves. PMID- 11725987 TI - The effect of long-term contact lens wear on the cells of the cornea. AB - PURPOSE: To review the effects of long-term contact lens wear on the cells of the cornea. METHODS: Published investigations into the effects of long-term contact lens wear on the corneal epithelium, keratocytes, and endothelium were reviewed. RESULTS: Results from multiple studies indicate that long-term daily wear of contact lenses causes endothelial polymegethism but has no effect on corneal epithelium (thickness, surface cell size); keratocytes (density, stromal reflectivity); or endothelial cell density, permeability, or pump rate. Extended wear also causes no endothelial changes other than polymegethism, but it affects the epithelium (decreased thickness, increased surface cell size) and keratocytes (possible decreased density). CONCLUSIONS: Investigators have not found detrimental effects on the cells of the cornea from the long-term use of daily wear contact lenses. Although contact lenses cause endothelial polymegethism, no functional deficits ensue. Extended wear lenses may cause changes in all three cell types, but it is not known if these effects are detrimental nor if they occur with newer lenses of higher oxygen transmissibility. Patients can be reassured that modern contact lenses can be worn for many years in daily wear fashion (and possibly in extended wear with lenses of very high oxygen transmissibility) without damaging the cells of the cornea. PMID- 11725988 TI - Epithelial split associated with wear of a silicone hydrogel contact lens. AB - PURPOSE: To describe the appearance and management of a superior epithelial arcuate lesion (SEAL) observed in association with the wear of a silicone hydrogel lens. METHODS: We present a case of a 30-year-old male who presented with a SEAL after 4 months of wear of a silicone hydrogel lens on a 30-day extended wear schedule. Bacteriological examination of the lenses and ocular adnexa was conducted at the time of the event. The clinical presentation and management are presented and compared to a review of cases associated with hydrogel lenses. RESULTS: The patient presented with symptoms of mild irritation. A linear splitwas noted in the superior peripheral cornea of the affected eye and was associated with limbal injection, stromal infiltrates, and fluorescein staining. Normal ocular biota was recovered from the lenses and eyelids of both eyes. Visual acuity remained unchanged and the lesion resolved following prophylactic treatment and discontinuation of lens wear for 8 days. CONCLUSIONS: The diagnosis and management of this SEAL case in a silicone hydrogel lens wearer mirrored that of previous events associated with hydrogel lenses. The possible etiologies of SEAL in this case are discussed and include mechanical irritation and dehydration of the lens surface. PMID- 11725989 TI - Long-term follow-up of corneal graft survival following bone marrow transplantation in the Maroteaux-Lamy syndrome. AB - PURPOSE: To present a case of Maroteaux-Lamy syndrome (MLS, mucopolysaccharidosis [MPS] type VI) who underwent bone marrow transplantation (BMT) for gene transfer at the age of 13, and penetrating keratoplasty at the age of 17, and maintained clear corneal grafts bilaterally for 13 years. To our knowledge, this is the longest follow-up reported on corneal graft survival in a patient with MLS and BMT. METHODS: In 1982, BMT was successfully performed on a 13-year-old girl with MLS with growth retardation, typical facial features, skeletal and joint deformities, hepatosplenomegaly, cardiopulmonary dysfunction, and corneal clouding. Corneal transplantation was done on the left eye in 1986, and on the right eye in 1987 (6 months later) without difficulty or complication. RESULTS: Thirteen years postoperatively, the patient was systemically well, and both eyes retained clear corneal grafts. CONCLUSION: BMT retarded further dysfunction from MLS, and the corneal transplants retained clarity. Further controlled studies with longer follow-up are required to establish the efficacy of BMT in ocular manifestations of MPS or MLS. PMID- 11725990 TI - Funding for research in nursing education. PMID- 11725991 TI - Empowering versus enabling in academia. AB - The literature endorses empowered nurses as essential to changing not only the nursing profession but also the current health care system. Nurse educators have embraced the concept of empowering students. However, faculty frequently enable students rather than empower them in both clinical and classroom settings. This article describes the difference between enabling and empowering. Collegiality, communication, accountability, and autonomy are components of a model used to compare empowering versus enabling in the academic setting. Nurse educators will become aware of behaviors that sabotage the empowerment of students. Strategies designed to facilitate students' development of empowering behaviors will be provided throughout the article. PMID- 11725992 TI - Student and faculty perceptions of effective clinical instructors in ADN programs. AB - The purpose of this study was to identify the perceptions of associate degree nursing (ADN) students and faculty of characteristics of effective clinical teachers and determine whether there were differences between these two groups. A survey was conducted of 292 students in various levels of their ADN programs and 59 faculty members from the same five programs, which were randomly selected from across Michigan. Data were collected using the Nursing Clinical Effectiveness Inventory, which includes 48 characteristics of effective clinical instructors arranged in five subscales. Students identified "demonstrates clinical skills and judgment" as the most important characteristic of effective clinical instructors, while faculty identified "explains clearly" as the most important characteristic. There was agreement on 6 of the top 10 characteristics identified by both groups. Both groups rated "directs student to useful literature in nursing" as the least important characteristic of effective clinical instructors. The students' and faculty's perceptions of effective clinical instructors differed by subscales, with students identifying evaluation characteristics as most important (mean = 4.73, SD = .42) and faculty identifying interpersonal relationships as most important (mean = 4.72, SD = .31). A t test indicated a significant difference between student and faculty means for the interpersonal relationships subscales, with faculty rating this group of characteristics as more important than students did (t = 2.49, p = .0 14). PMID- 11725993 TI - Influence of a persuasive strategy on nursing students' beliefs and attitudes toward provision of care to people living with HIV/AIDS. AB - Based on the theory of planned behavior and the elaboration likelihood model, the aim of this study was to verify the effect of persuasive messages on nursing students' beliefs and attitudes regarding provision of care to people living with HIV/AIDS. The assumption was that a persuasive communication strategy induces a constructive change in beliefs and attitudes regarding provision of care. Baseline data collection was performed among a group of 74 nursing students (experimental group = 27; control group = 47). The questionnaire assessed the variables of the theory of planned behavior (i.e., beliefs, attitudes, perceived behavioral control). The results confirmed that persuasive messages generated a change in beliefs and attitudes of the nursing students concerning providing care to people living with HIV/AIDS. It appears that this strategy of modifying behavioral predispositions is effective and generates cognitive and affective changes. Therefore, educational programs should take these observations into consideration to ensure that future nurses are better prepared to provide appropriate care to people living with HIV/AIDS. PMID- 11725994 TI - Implementing a completely Web-based nursing research course: instructional design, process, and evaluation considerations. PMID- 11725995 TI - Collaboration for teaching innovation: writing across the curriculum in a school of nursing. PMID- 11725996 TI - First Aid and Health Kamp for Kids: combining learning with service. PMID- 11725997 TI - Experiential learning in drug and alcohol education. PMID- 11725998 TI - Peer instruction in the learning laboratory: a strategy to decrease student anxiety. PMID- 11725999 TI - Role strain in male diploma nursing students: a descriptive quantitative study. PMID- 11726000 TI - Hypersensitivity reactions during therapy with the nucleoside reverse transcriptase inhibitor abacavir. AB - BACKGROUND: Hypersensitivity reactions consist of a variable group of clinical findings and have been described for a wide variety of chemical compounds. OBJECTIVE: This review characterizes the clinical profile of hypersensitivity to the nucleoside reverse transcriptase inhibitor abacavir sulfate. METHODS: We performed a retrospective medical review of pooled adverse events data from approximately 200,000 patients who received abacavir in clinical trials, through expanded-access programs, or by prescription from 1996 through 2000. Screened cases of hypersensitivity were classified as either definitive or probable. Definitive cases were identified when initial symptoms resolved on interruption of abacavir therapy and returned on reintroduction of abacavir therapy. RESULTS: A total of 1803 cases were identified, 1302 in the 30,595 patients participating in clinical trials or the expanded-access program and 501 in patients from the post-marketing experience. On review, 176 (9.8%) of these cases were considered definitive and the remainder probable. Based on the 1302 cases identified in clinical trials or the expanded-access program, the calculated incidence of hypersensitivity was 4.3%. Symptoms reported in > or = 20% of cases of this multiorgan reaction included fever, rash, malaise/fatigue, and gastrointestinal symptoms such as nausea, vomiting, and diarrhea, among others. Respiratory symptoms occurred in 30% of cases and included dyspnea (12%), cough (10%), and pharyngitis (6%). In 90% of cases, hypersensitivity reactions occurred within the first 6 weeks after initiation of abacavir (median time, 11 days); after an initial reaction, rechallenge with abacavir resulted in the reappearance of symptoms within hours of reexposure. Hypotension was present in 25% of these rechallenge reactions. Among patients who received abacavir in clinical trials, the mortality rate was 0.03% (3 per 10,000 patients). CONCLUSIONS: Hypersensitivity to abacavir is an idiosyncratic reaction and a distinct clinical syndrome characterized predominantly by systemic involvement. It can be expected to appear as a treatment-limiting event in approximately 5% of patients. The appearance of clinical symptoms consistent with this syndrome mandates immediate discontinuation of abacavir. Hypersensitivity to abacavir is an absolute contraindication to subsequent treatment with any formulation that includes this agent. PMID- 11726001 TI - Respiratory illness caused by picornavirus infection: a review of clinical outcomes. AB - BACKGROUND: Respiratory infections result from invasion of the respiratory tract, mainly by viruses, and are the leading cause of acute morbidity in individuals of all ages worldwide. During peak season, picornaviruses cause 82% of all episodes of acute nasopharyngitis (the common cold), the most frequent manifestation of acute respiratory infection, and produce more restriction of activity and physician consultations annually than any other viral or bacterial source of respiratory illness. OBJECTIVE: This article reviews the clinical impact and outcomes of picornavirus-induced respiratory infections in specific populations at risk for complications. It also discusses the potential economic impact of the morbidity associated with picornavirus-induced respiratory infection. METHODS: Relevant literature was identified through searches of MEDLINE, OVID, International Pharmaceutical Abstracts, and Lexis-Nexis. The search terms used were picornavirus, rhinovirus, enterovirus, viral respiratory infection, upper respiratory infection, disease burden, economic, cost, complications, asthma, COPD, immunocompromised, elderly otitis media, and sinusitis. Additional publications were identified from the reference lists of the retrieved articles. CONCLUSIONS: Based on the clinical literature, picornavirus infections are associated with severe morbidity as well as considerable economic and societal costs. Future research should focus on identifying patterns of illness and the costs associated with management of these infections. New treatments should be assessed not only in terms of their ability to produce the desired clinical outcome, but also in terms of their ability to reduce the burden of disease, decrease health care costs, and improve productivity. PMID- 11726002 TI - Considerations for optimal iron use for anemia due to chronic kidney disease. AB - BACKGROUND: Availability of recombinant human erythropoietin (rHuEPO) has improved the treatment of anemia due to chronic kidney disease (CKD). Iron deficiency is the most common cause of resistance to rHuEPO therapy, contributing to ineffective erythropoiesis and hematocrit/hemoglobin values below the recommended target range (33%-36%/11-12 g/dL). I.v. iron supplementation is necessary to meet increased iron demands from stimulation of erythropoiesis and chronic blood loss; however, questions remain as to the optimal supplementation strategy to maintain appropriate yet safe iron status. Treatment guidelines for anemia management have been developed through the National Kidney Foundation Kidney Disease Outcomes Quality Initiative (NKF-K/DOQI). OBJECTIVE: This review presents the basis of need for the NKF-K/DOQI guidelines and includes detailed information concerning iron physiology, metabolism, iron preparations, and evaluation of iron status. METHODS: This review was based on a MEDLINE search and complemented by references from the NKF-K/DOQI guidelines (whose review extended beyond MEDLINE). References focusing on normal iron physiology and metabolism, alterations in iron physiology in patients with CKD, laboratory evaluation methods, and strategies for iron supplementation were obtained from MEDLINE and reviewed for content. RESULTS: Controversy over appropriate use of iron supplementation has led to disparity in accepted practice procedures. Oral iron (ferrous salts and polysaccharide iron complex) and i.v. iron preparations (iron dextran, sodium ferric gluconate, and iron sucrose) are available. Problems with oral iron supplementation include limited absorption and patient noncompliance. Although most available data on i.v. iron use in the United States are specific to iron dextran preparations, published information based on clinical use of sodium ferric gluconate and iron sucrose products has been promising. The use of chronic i.v. iron administration to sustain iron stores has been more widely accepted to prevent development of absolute and functional iron deficiency. CONCLUSIONS: Although iron therapy is commonly warranted in patients with CKD, questions remain as to the most favorable supplementation strategy to optimize therapy through improvements in hematocrits, efficient use of rHuEPO, and maintenance of appropriate and safe iron levels. Clinicians will need to devise strategies based on the compilation of information from clinical experience and the available literature. Clinical practice guidelines devised by the NKF-K/DOQI have provided a useful tool for the medical community using both these resources. PMID- 11726003 TI - Prevalence of treated dry eye disease in a managed care population. AB - BACKGROUND: Survey studies are of limited utility in estimating the prevalence of treated dry eye. The use of claims data, which include only individuals who have a diagnosis of the disorder, provides a better estimation of the clinical significance of dry eye symptoms and appraisal of community needs. OBJECTIVE: The purpose of this paper is to estimate the prevalence of treated dry eye disease using a nonsurvey methodology. METHODS: Patients with dry eye diagnoses or who underwent punctal occlusion procedures were identified from PharMetrics' Integrated Outcomes database of medical claims for approximately 10 million patients enrolled in managed care plans. Prevalence estimates were calculated for 1997 and 1998. RESULTS: The prevalence of dry eye was 0.48% in 1997 and 0.39% in 1998, representing 25,180 and 27,289 cases, respectively. Patients aged > or = 65 years were approximately 4 times as likely as those aged < 65 years to be diagnosed with keratoconjunctivitis sicca or tear film insufficiency. In 1997, dry eye was diagnosed or treated in 0.65% of women compared with 0.26% of men (P < 0.001). Rates of dry eye disease in 1998 were highest among women aged 75 to 79 years (2.02%) and men aged 80 to 84 years (1.30%). Women tended to receive a diagnosis at a younger age than did men. The most common diagnosis was tear film insufficiency (73.96% and 73.41% of dry eye patients in 1997 and 1998, respectively). The most common procedure was lacrimal punctal occlusion by plug (7.78% and 8.74% of dry eye patients in 1997 and 1998, respectively). CONCLUSION: The prevalence of treated dry eye disease is 0.4% to 0.5% and is highest among women and the elderly. PMID- 11726004 TI - Diagnosis and treatment of upper respiratory tract infections in the primary care setting. AB - BACKGROUND: Acute respiratory tract infections such as acute exacerbations of chronic bronchitis (AECB), acute otitis media (AOM), and acute bacterial rhinosinusitis (ABRS) account for approximately 75% of antibiotic prescriptions written and are among the leading reasons for physician office visits in the United States. Resistance of the predominant pathogens in respiratory tract infections (Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis) to available antibiotics has led clinicians to reevaluate the diagnosis and management of these infections. OBJECTIVE: The purpose of this review is to provide primary care practitioners with an accessible combined resource for the management of AECB, AOM, and ABRS. METHODS: This review was based on discussions from a roundtable meeting (sponsored by an educational grant from GlaxoSmithKline) that convened clinicians versed in the management of upper and lower respiratory tract infections. In addition, primary articles were identified by a MEDLINE search and through secondary sources. RESULTS: To reduce the prevalence of resistance, judicious and appropriate use of antibiotics must be implemented in clinical practice. With accurate diagnosis of bacterial and nonbacterial conditions, and patient education on antibiotic use and misuse, the excessive use of antibiotics and ensuing resistance can be reduced. The incorporation of pharmacokinetic and pharmacodynamic data with minimum inhibitory concentration values can provide a more comprehensive assessment of antibiotic activity in vivo. Stratification of patients with AECB according to patient characteristics and frequency of exacerbation can be used to determine which patients will benefit from antibiotic treatment and to guide clinicians in their choice of antibiotic. The Drug-Resistant Streptococcus pneumoniae Therapeutic Working Group has issued recommendations on the management of AOM based on prior antibiotic therapy, which is a risk factor for antimicrobial resistance. The Sinus and Allergy Health Partnership guidelines for the treatment of ABRS in adults and children are based on the predicted efficacy of various antibiotics as well as patient age, severity of disease, likelihood of bacterial infection, likelihood of spontaneous resolution, and in vitro susceptibility of the predominant pathogens based on pharmacokinetic and pharmacodynamic breakpoints. CONCLUSIONS: Guidelines for the management of AECB, AOM, and ABRS emphasize the importance of differentiating between bacterial and nonbacterial infections, choosing an antibiotic based on the likelihood of infection with resistant pathogens, and providing coverage against the predominant pathogens. The judicious use of antibiotics also has been identified as an instrumental part of controlling unnecessary antibiotic use and subsequent resistance. PMID- 11726005 TI - Relationship between patient self-assessment of erectile dysfunction and the sexual health inventory for men. AB - BACKGROUND: The Sexual Health Inventory for Men (SHIM) has been shown to possess favorable statistical properties in diagnosing the presence and severity of erectile dysfunction (ED). However, the SHIM has not been compared with patient self-assessment of ED. OBJECTIVE: This article describes an independent validation study examining the correlation and agreement between the SHIM and patient self-assessment of ED with respect to the severity of ED at baseline and after treatment, and in terms of change from baseline. METHODS: The study population consisted of 247 male outpatients with ED participating in a multicenter, double-blind, placebo-controlled, flexible-dose (25-100 mg/d) Phase IIIb clinical trial in which they were randomized equally to sildenafil citrate or placebo. Patients assessed their degree of ED as severe, moderate, minimal/mild, or no problem at baseline and after 12 weeks of treatment. They also responded to the 5 questions on the SHIM, after which their degree of ED was calculated based on the SHIM total score. RESULTS: In general, the SHIM and the single-item self-assessment question produced similar descriptive profiles of the severity of ED. Kendall tau-b correlations were 0.66 (95% CI, 0.58-0.74) at baseline, 0.86 (95% CI, 0.82-0.90) after treatment, and 0.72 (95% CI, 0.67-0.77) for change from baseline. Agreement between instruments, measured by the weighted kappa statistic, mirrored the correlations at baseline and after treatment. As expected, both measures correlated moderately with improvement in erections and treatment satisfaction of both patient and partner. CONCLUSION: The moderate-to high correlation and agreement between the SHIM and patient self-assessment of ED validate the SHIM for use in the diagnostic classification of ED severity. PMID- 11726006 TI - Case reports of the reemergence of psychotic symptoms after conversion from brand name clozapine to a generic formulation. AB - BACKGROUND: The use of generic drugs has resulted in considerable cost savings; however, whether all generics are truly bioequivalent to their brand-name counterparts is questionable. Although the efficacy of clozapine in the management of treatment-resistant schizophrenia has been well established, reports of relapse after conversion to a generic formulation are becoming more common. OBJECTIVE: This article presents 7 case studies of patients in a long term residential care facility who experienced a relapse of psychotic symptoms when the pharmacy inadvertently switched their therapy from brand-name clozapine to a generic formulation. Neither patients, physicians, nor staff of the facility were aware of this switch. Possible reasons for the apparent increased risk of relapse in some patients switched to the generic formulation of clozapine are explored, with reference to US Food and Drug Administration bioequivalence standards and reports. RESULTS: All 7 patients, whose condition had been well stabilized with brand-name clozapine, experienced a rapid and profound deterioration after the switch to the generic formulation. Five patients required hospitalization. All patients responded well when brand-name clozapine was reinstated. CONCLUSION: The findings suggest that brand-name clozapine and the generic formulation may display important clinical differences, and a comparable therapeutic response may not be achievable despite adequate monitoring. Large, controlled, prospective trials are needed to clarify the potential for treatment failure with the use of generic clozapine. PMID- 11726008 TI - A welcome debate on statistical analysis. PMID- 11726007 TI - Glycemic control with Humalog Mix25 in type 2 diabetes inadequately controlled with glyburide. AB - BACKGROUND: Humalog Mix25 (Mix25) is a premixed insulin mixture of 25% insulin lispro and 75% neutral protamine lispro. OBJECTIVE: The aim of this study was to quantitate the improvement in glycemic control achieved with Mix25 versus the maximum dose of glyburide (GB) in patients with type 2 diabetes inadequately controlled with GB. METHODS: In this randomized, parallel, open-label comparative study, patients with type 2 diabetes received either Mix25 before the morning and evening meals for 4 months or GB 15 mg daily for 4 months. Glycemic control was assessed by glycosylated hemoglobin (HbA1c) measurements, 4-point self-monitored blood glucose profiles, and patient-reported hypoglycemia. Patients also completed a treatment satisfaction questionnaire at the end of the study. RESULTS: All 172 patients were white; 85 were randomized to receive Mix25. The mean age was 59.5 +/- 8.2 years, and 35.5% (61/172) were men. The mean body mass index was 27.2 kg/m2. The mean duration of type 2 diabetes was 10.2 +/- 6.6 years, and the mean duration of sulfonylurea treatment was 5.8 +/- 5.9 years. The mean HbA1c and fasting blood glucose levels were 10.07% +/- 1.4% and 11.6 +/- 2.8 mmol/L, respectively, in the glyburide group and 9.85% +/- 1.2% and 12.2 +/- 2.9 mmol/L, respectively, in the Mix25 group. There were no statistically significant differences between the treatment groups at baseline for any of the demographic or efficacy variables. At end point, mean HbA1c was significantly lower in the Mix25 group than in the GB group (Mix25, 8.5% +/- 1.3%; GB, 9.4% +/- 1.8%; P = 0.001). A larger decrease from baseline in HbA1c and in all self-monitored blood glucose values was observed in the Mix25 group: -1.4% versus -0.7% for HbA1c, P = 0.004; -2.8 mmol/L versus -1.1 mmol/L for fasting blood glucose, P < 0.01; -5.1 mmol/L versus -1.7 mmol/L for the morning 2-hour postprandial blood glucose, P < 0.001; -2.2 mmol/L versus -0.8 mmol/L for the evening preprandial blood glucose, P < 0.05; and -4.4 mmol/L versus -1.5 mmol/L for the evening 2-hour postprandial blood glucose, P < 0.001. Patients expressed more satisfaction with Mix25 than with GB, as measured by the weighted combined score on a treatment satisfaction questionnaire (2.0 +/- 1.3 vs 0.7 +/- 1.3). The mean hypoglycemia rate (events per patient per 30 days) was significantly higher in the Mix25 group at end point (Mix25, 0.30 +/- 0.53; GB, 0.05 +/- 0.20; P < 0.001). CONCLUSIONS: Compared with maximum-dose GB, twice-daily injections of Mix25 resulted in improved glycemic control and treatment satisfaction, and were associated with a predictably higher rate of hypoglycemia in this group of patients with type 2 diabetes who were inadequately controlled with maximum-dose GB. Although the inclusion of patients who were inadequately controlled with GB was intended to allow a comparison of the 2 treatments with respect to efficacy and tolerability in a real-life setting, a double-blind comparison in treatment-naive individuals may have resulted in a different outcome. PMID- 11726009 TI - Selection of clinical, patient-reported, and economic end points in acute exacerbation of chronic bronchitis. AB - BACKGROUND: Acute exacerbation of chronic bronchitis (AECB) places tremendous burden on patients, providers, employers, and health care systems. OBJECTIVE: The purpose of this paper is to (1) review the clinical, patient-reported, and economic measures used to evaluate disease burden and treatment effectiveness in AECB in clinical trials and (2) propose a guide for selecting study end points in AECB that will help capture all the relevant disease outcomes. METHODS: Two literature searches of the PubMed database were conducted to identify studies of clinical trials in bronchitis and evaluate the clinical, patient-reported, and economic end points used in these studies. RESULTS: Previous studies have focused primarily on clinician-assessed outcomes, which do not capture the full impact of AECB on patients' lives. Reporting mechanisms for most end points have been inconsistent, limiting the ability to compare information or interpret differences. Previous studies have given limited attention to patient-reported outcomes and the economic implications of AECB. Patient-reported outcomes such as speed of symptom relief and work productivity are important parameters for assessing treatment effectiveness and provide practical information for treatment evaluation. CONCLUSIONS: Additional research is needed to develop, examine, and validate patient-reported outcomes and the indirect costs of AECB. Measuring the relevant clinical, economic, and patient-reported outcomes in AECB patients using standardized methods may lead to a clearer understanding of the disease burden and the role, effectiveness, and cost-effectiveness of antibiotic treatment. PMID- 11726010 TI - Assessment of the association between blood pressure control and health care resource use. AB - BACKGROUND: Several studies have reported increased health care resource use among hypertensive patients with uncontrolled blood pressure (BP). OBJECTIVE: The purpose of this study was to investigate the relationship between BP control and health care resource use. METHODS: Data were obtained from the Caring for Hypertension on Initiation: Costs and Effectiveness (CHOICE) study, a multicenter feasibility study of actual physician and patient behavior and clinical outcomes in a naturalistic setting. Adult patients with newly diagnosed hypertension were randomized to either Group 1 (treatment with diuretics or beta-blockers) or Group 2 (treatment with calcium channel blockers or angiotensin-converting enzyme inhibitors) and followed for 5 +/- 1 months. Physicians practiced standard care while documenting medications, BP measurements, and health care resource use for their patients. A subsequent analysis evaluating the relationship between BP and physician visits was performed for the whole population and for a subpopulation of patients with at least 4 months of follow-up data. Cox regression was used to model time to next visit. RESULTS: A total of 512 patients with newly diagnosed hypertension were followed: 399 had follow-up data for at least 4 months. Baseline demographic characteristics were similar in the 2 groups. Kaplan-Meier curves and a log-rank test showed that the time to next visit for patients with uncontrolled BP was significantly shorter than for patients whose BP was controlled (P < 0.05). On average, patients with uncontrolled BP (> or = 140/90 mm Hg) had follow-up office visits approximately 13 days earlier than patients with controlled BP (< 140/90 mm Hg). This association remained significant after adjustment for repeated measures, and after exclusion of the first return visit. Cox regression analysis showed that higher systolic and diastolic BP measurements were significantly associated with a shorter time to next visit, after adjustment for age and sex. Total estimated costs during the study period were $170 per patient for medications and $283 per patient for office visits. CONCLUSIONS: In the CHOICE study, higher BP was associated with a shorter time to next visit. Office visits were the main cost driver in the short-term management of hypertension. PMID- 11726011 TI - Dealing with skewed cost data. PMID- 11726012 TI - The changing character of consumer and environmental regulation: a comparison of Europe and the United States. PMID- 11726013 TI - Sensitivity analysis, Monte Carlo risk analysis, and Bayesian uncertainty assessment. AB - Standard statistical methods understate the uncertainty one should attach to effect estimates obtained from observational data. Among the methods used to address this problem are sensitivity analysis, Monte Carlo risk analysis (MCRA), and Bayesian uncertainty assessment. Estimates from MCRAs have been presented as if they were valid frequentist or Bayesian results, but examples show that they need not be either in actual applications. It is concluded that both sensitivity analyses and MCRA should begin with the same type of prior specification effort as Bayesian analysis. PMID- 11726014 TI - Human interindividual variability in susceptibility to airborne particles. AB - Part of the explanation for the persistent epidemiological findings of associations between mortality and morbidity with relatively modest ambient exposures to airborne particles may be that some people are much more susceptible to particle-induced responses than others. This study assembled a database of quantitative observations of interindividual variability in pharmacokinetic and pharmacodynamic parameters likely to affect particle response. The pharmacodynamic responses studied included data drawn from epidemiologic studies of doses of methacholine, flour dust, and other agents that induce acute changes in lung function. In general, the amount of interindividual variability in several of these pharmacodynamic response parameters was greater than the variability in pharmacokinetic (breathing rate, deposition, and clearance) parameters. Quantitatively the results indicated that human interindividual variability of breathing rates and major pharmacokinetic parameters-total deposition and tracheobronchial clearance-were in the region of Log(GSD) = 0.1 to 0.2 (corresponding to geometric standard deviations of 10(.1)-10(.2) or 1.26 1.58). Deposition to the deep lung (alveolar region) appeared to be somewhat more variable: Log(GSD) of about 0.3 (GSD of about 2). Among pharmacodynamic parameters, changes in FEV1 in response to ozone and metabisulfite (an agent that is said to act primarily on neural receptors in the lung) were in the region of Log(GSD) of 0.2 to 0.4. However, similar responses to methacholine, an agent that acts on smooth muscle, seemed to have still more variability (0.4 to somewhat over 1.0, depending on the type of population studied). Similarly high values were suggested for particulate allergens. Central estimates of this kind of variability, and the close correspondence of the data to lognormal distributions, indicate that 99.9th percentile individuals are likely to respond at doses that are 150 to 450-fold less than would be needed in median individuals. It seems plausible that acute responses with this amount of variability could form part of the mechanistic basis for epidemiological observations of enhanced mortality in relation to ambient exposures to fine particles. PMID- 11726015 TI - Using epidemiological studies to check the consistency of the cancer risks predicted by high-dose animal experiments: a methodological review. AB - Epidemiological studies have been cited in the literature as evidence both for and against the human cancer risks predicted by high-exposure rodent studies. However, there has been little overall consistency in the ways that these animal to-human comparisons have been made. This review examines some examples of these types of comparisons and describes the methods and techniques used by different investigators. Eleven "key decision areas" that need to be addressed are identified and recommendations for consistent, logical, and statistically appropriate approaches that might be taken to standardize the process are provided. In general, it is suggested that investigators provide the most useful information when they use logical, transparent, and statistically valid comparisons to pursue limited and focused objectives, such as directly testing the validity of an existing regulatory guidance value. Other recommendations include selecting biologically plausible extrapolative models that fit the data and drawing conclusions that are consistent with the study results and objectives. PMID- 11726016 TI - Likelihood contour method for the calculation of asymptotic upper confidence limits on the risk function for quantitative responses. AB - This article develops a computationally and analytically convenient form of the profile likelihood method for obtaining one-sided confidence limits on scalar valued functions phi = phi(psi) of the parameters psi in a multiparameter statistical model. We refer to this formulation as the likelihood contour method (LCM). In general, the LCM procedure requires iterative solution of a system of nonlinear equations, and good starting values are critical because the equations have at least two solutions corresponding to the upper and lower confidence limits. We replace the LCM equations by the lowest order terms in their asymptotic expansions. The resulting equations can be solved explicitly and have exactly two solutions that are used as starting values for obtaining the respective confidence limits from the LCM equations. This article also addresses the problem of obtaining upper confidence limits for the risk function in a dose response model in which responses are normally distributed. Because of normality, considerable analytic simplification is possible and solution of the LCM equations reduces to an easy one-dimensional root-finding problem. Simulation is used to study the small-sample coverage of the resulting confidence limits. PMID- 11726017 TI - Limitations to empirical extrapolation studies: the case of BMD ratios. AB - Extrapolation relationships are of keen interest to chemical risk assessment in which they play a prominent role in translating experimentally derived (usually in animals) toxicity estimates into estimates more relevant to human populations. A standard approach for characterizing each extrapolation relies on ratios of pre existing toxicity estimates. Applications of this "ratio approach" have overlooked several sources of error. This article examines the case of ratios of benchmark doses, trying to better understand their informativeness. The approach involves mathematically modeling the process by which the ratios are generated in practice. Both closed form and simulation-based models of this "data-generating process" (DGP) are developed, paying special attention to the influence of experimental design. The results show the potential for significant limits to informativeness, and revealing dependencies. Future applications of the ratio approach should take imprecision and bias into account. Bootstrap techniques are recommended for gauging imprecision, but more complicated techniques will be required for gauging bias (and capturing dependencies). Strategies for mitigating the errors are suggested. PMID- 11726018 TI - Comparison of the EU T25 single point estimate method with benchmark dose response modeling for estimating potency of carcinogens. AB - The T25 single-point estimate method of evaluating the carcinogenic potency of a chemical, which is currently used by the European Union (EU) and is denoted the EU approach, is based on the selection of a single dose in a chronic bioassay with an incidence rate that is significantly higher than the background rate. The T25 is determined from that single point by a linear extrapolation or interpolation to the chronic dose (in mg/kg/day), at which a 25% increase in the incidence of the specified tumor type is expected, corrected for the background rate. Another method used to obtain a carcinogenic potency value based on a 25% increase in incidence above the background rate is the estimation of a T25 derived from a benchmark dose (BMD) response model fit to the chronic bioassay data for the specified tumor type. A comparison was made between these two methods using 276 chronic bioassays conducted by the National Toxicology Program. In each of the 2-year bioassays, a tumor type was selected based on statistical and biological significance, and both EU T25 and BMD T25 estimates were determined for that end point. In addition, simulations were done using underlying cumulative probability distributions to examine the effect of dose spacing, the number of animals per dose group, the possibility of a dose threshold, and variation in the background incidence rates on the EU T25 and BMD estimates. The simulations showed that in the majority of cases the EU T25 method underestimated the true T25 dose and overestimated the carcinogenic potency. The BMD estimate is generally less biased and has less variation about the true T25 value than the EU estimate. PMID- 11726019 TI - Estimation of tuberculosis risk and incidence under upper room ultraviolet germicidal irradiation in a waiting room in a hypothetical scenario. AB - Environmental control measures (ventilation, high-efficiency particulate air filtration, and upper room ultraviolet germicidal irradiation [UVGI]) are recommended to effectively control tuberculosis (TB) transmission from unsuspected TB patients in high-risk settings, but the effectiveness of their use is not often clear. This study presents a simulation model for a hypothetical hospital waiting room, in which the number of susceptible immunocompetent people in the waiting room follows a Poisson distribution (M = 5) in either low (annual number of TB patients = 5) or high TB risk settings (annual number of TB patients = 50), and used the model to evaluate the reduction of TB transmission risk by upper room UVGI. An exponential dose-response model was used for TB transmission and a two-zone model was used for evaluating the effect of upper room UVGI. Upper room UVGI reduced TB risk by 1.6-fold at 3 microW/cm2 UV irradiance in the upper room in the low TB risk setting and by 4.1-fold at 15 microW/cm2 UV irradiance in the upper room in the high TB risk setting. Use of upper room UVGI also reduced the mean annual new infection rate from 2.2 to 1.3 infections per year at 3 microW/cm2 and to 0.6 infections per year at 15 microW/cm2 in our hypothetical high-risk settings. The effect of upper room UVGI was sensitive to both vertical air velocity (air mixing) and UV irradiance level. Results from partitioning variability indicate that most variability of TB transmission risk came from waiting time in our hypothetical hospital. PMID- 11726020 TI - Stochastic estimates of exposure and cancer risk from carbon tetrachloride released to the air from the rocky flats plant. AB - Carbon tetrachloride is a degreasing agent that was used at the Rocky Flats Plant (RFP) in Colorado to clean product components and equipment. The chemical is considered a volatile organic compound and a probable human carcinogen. During the time the plant operated (1953-1989), most of the carbon tetrachloride was released to the atmosphere through building exhaust ducts. A smaller amount was released to the air via evaporation from open-air burn pits and ground-surface discharge points. Airborne releases from the plant were conservatively estimated to be equivalent to the amount of carbon tetrachloride consumed annually by the plant, which was estimated to be between 3.6 and 180 Mg per year. This assumption was supported by calculations that showed that most of the carbon tetrachloride discharged to the ground surface would subsequently be released to the atmosphere. Atmospheric transport of carbon tetrachloride from the plant to the surrounding community was estimated using a Gaussian Puff dispersion model (RATCHET). Time-integrated concentrations were estimated for nine hypothetical but realistic exposure scenarios that considered variation in lifestyle, location, age, and gender. Uncertainty distributions were developed for cancer slope factors and atmospheric dispersion factors. These uncertainties were propagated through to the final risk estimate using Monte Carlo techniques. The geometric mean risk estimates varied from 5.2 x 10(-6) for a hypothetical rancher or laborer working near the RFP to 3.4 x 10(-9) for an infant scenario. The distribution of incremental lifetime cancer incidence risk for the hypothetical rancher was between 1.3 x 10(-6) (5% value) and 2.1 x 10(-5) (95% value). These estimates are similar to or exceed estimated cancer risks posed by releases of radionuclides from the site. PMID- 11726021 TI - Reproductive and developmental risks from ethylene oxide: a probabilistic characterization of possible regulatory thresholds. AB - Ethylene oxide is a gas produced in large quantities in the United States that is used primarily as a chemical intermediate in the production of ethylene glycol, propylene glycol, non-ionic surfactants, ethanolamines, glycol ethers, and other chemicals. It has been well established that ethylene oxide can induce cancer, genetic, reproductive and developmental, and acute health effects in animals. The U.S. Environmental Protection Agency is currently developing both a cancer potency factor and a reference concentration (RfC) for ethylene oxide. This study used the rich database on the reproductive and developmental effects of ethylene oxide to develop a probabilistic characterization of possible regulatory thresholds for ethylene oxide. This analysis was based on the standard regulatory approach for noncancer risk assessment, but involved several innovative elements, such as: (1) the use of advanced statistical methods to account for correlations in developmental outcomes among littermates and allow for simultaneous control of covariates (such as litter size); (2) the application of a probabilistic approach for characterizing the uncertainty in extrapolating the animal results to humans; and (3) the use of a quantitative approach to account for the variation in heterogeneity among the human population. This article presents several classes of results, including: (1) probabilistic characterizations of ED10s for two quantal reproductive outcomes-resorption and fetal death, (2) probabilistic characterizations of one developmental outcome-the dose expected to yield a 5% reduction in fetal (or pup) weight, (3) estimates of the RfCs that would result from using these values in the standard regulatory approach for noncancer risk assessment, and (4) a probabilistic characterization of the level of ethylene oxide exposure that would be expected to yield a 1/1,000 increase in the risk of reproductive or developmental outcomes in exposed human populations. PMID- 11726022 TI - Short-Term dermal absorption and penetration of chemicals from aqueous solutions: theory and experiment. AB - Dermal penetration of organic chemical-contaminated water from showering and bathing scenarios is a concern of regulatory agencies that have been tasked with determining safe exposure levels. During household showering and bathing, nearly the entire surface area of the body is exposed for short periods of time (5-15 minutes). The primary means of predicting body burden during brief exposures is to estimate total chemical penetrated from the steady-state penetration rate using a skin permeability coefficient. A variety of approaches has been recommended to estimate "body burden." The purpose of this investigation was to collect experimental data from short-term exposures to an organic chemical (dibromomethane [DBM]) in aqueous solution so that methods for estimating body burden could be compared. Rat skins were exposed in vitro to saturated aqueous solutions of DBM for 20 minutes and the amount of chemical in the receptor solution and the skin was analyzed. The total DBM mass in the receptor solution and the skin was taken to represent an in vivo body burden. These results were compared with the estimates of penetration from steady-state calculations, square root of time calculations, and a biologically based mathematical model. Results indicated that the amount of chemical in the skin and its fate during short exposures is important. The square root of time approach predicted total amount of chemical absorbed and penetrated better than did the steady-state approach. The biologically based mathematical model accurately predicted total body burden and could be used to distinguish between the amount of chemical in the skin and the amount of chemical that penetrated through the skin, which would be useful for understanding local toxicity. PMID- 11726023 TI - Sexual risk compensation and HIV/STD transmission: empirical evidence and theoretical considerations. AB - This study was conducted to answer the question, "Are sexual risk behaviors subject to compensation?" For example, do people who increase their use of condoms compensate for this reduction in human immunodeficiency virus and sexually transmitted disease (HIV/STD) risk by engaging in more overall acts of intercourse or by having sex with more partners than before? Utilizing the HIV prevention literature, studies in which participants demonstrated sexual risk compensation were identified. A simple HIV/STD transmission model was applied to these data to determine whether compensation produced a net increase in HIV/STD risk, despite positive changes in one or more aspects of sexual behavior. Although a number of studies were found in which there were simultaneous increases in condom use and the overall number of acts of intercourse, in none of these instances was there an overall increase in HIV/STD risk. Moreover, none of these studies reported concomitant increases in the number of sex partners. Extensive modeling exercises also were conducted to determine the theoretical conditions under which compensation would produce a net increase in risk. The results of the modeling exercise indicated that relatively small increases in overall sexual activity could be sufficient to offset risk-reduction gains due to increased condom use in populations in which baseline condom use is very low. In sum, although sexual risk compensation occurs, no empirical evidence was found that this compensation is sufficient to offset reductions in risk due to greater condom use, despite the theoretical plausibility of this scenario. PMID- 11726024 TI - Environmental tobacco smoke revisited: the reliability of the data used for risk assessment. AB - Several epidemiological studies have found a weak, but consistent association between lung cancer in nonsmokers and exposure to environmental tobacco smoke (ETS). In addition, a purported link between such exposure and coronary heart disease (CHD) has been of major concern. Although it is biologically plausible that ETS has a contributory role in the induction of lung cancer in nonsmoking individuals, dose-response extrapolation-supported by the more solid database for active smokers-gives an additional risk for lung cancer risk that is more than one order of magnitude lower than that indicated by major positive epidemiological studies. The discrepancy between available epidemiological data and dosimetric estimates seems, to a major part, to reflect certain systematic biases in the former that are difficult to control by statistical analysis when dealing with risks of such low magnitudes. These include, most importantly, misclassification of smoking status, followed by inappropriate selection of controls, as well as certain confounding factors mainly related to lifestyle, and possibly also hereditary disposition. A significant part of an association between lung cancer and exposure to ETS would disappear, if, on the average, 1 patient out of 20 nonsmoking cases had failed to tell the interviewer that he had, in fact, recently stopped smoking. In the large International Agency for Research on Cancer (IARC) multicenter study even lower misclassification rates would abolish the weak, statistically nonsignificant associations that were found. In the former study an apparent significant protective effect from exposure to ETS in childhood with respect to lung cancer later in life was reported, a most surprising finding. The fact that the mutation spectrum of the p53 tumor suppressor gene in lung tumors of ETS-exposed nonsmokers generally differs from that found in tumors of active smokers lends additional support to the notion that the majority of tumors found in ETS-exposed nonsmokers have nothing to do with tobacco smoke. The one-sided preoccupation with ETS as a causative factor of lung cancer in nonsmokers may seriously hinder the elucidation of the multifactorial etiology of these tumors. Due to the high prevalence of cardiovascular disease in the population, even a modest causal association with ETS would, if valid, constitute a serious public health problem. By pooling data from 20 published studies on ETS and heart disease, some of which reported higher risks than is known to be caused by active smoking, a statistically significant association with spousal smoking is obtained. However, in most of these studies, many of the most common confounding risk factors were ignored and there appears to be insufficient evidence to support an association between exposure to ETS and CHD. Further, it seems highly improbable that exposure to a concentration of tobacco smoke at a level that is generally much less than 1% of that inhaled by a smoker could result in an excess risk for CHD that-as has been claimed-is some 30% to 50% of that found in active smokers. There are certainly valid reasons to limit exposure to ETS as well as to other air pollutants in places such as offices and homes in order to improve indoor air quality. This goal can be achieved, however, without the introduction of an extremist legislation based on a negligible risk of lung cancer as well as an unsupported and highly hypothetical risk for CHD. PMID- 11726025 TI - Stochastic simulation of catastrophic hail and windstorm indemnities in the Dutch greenhouse sector. AB - A risk analysis was performed to examine the effect of changes in the Dutch greenhouse sector on the probability of occurrence and magnitude of indemnities induced by catastrophic natural hazards. Analyzed historical indemnities, which included direct and consequential losses resulting from severe hail and windstorms, were used as input in a stochastic simulation model. Applications of the stochastic simulation model were illustrated under alternative risk conditions. A comparison was made between the current structure of greenhouse production and the expected structure in the next decade that differs with respect to the spatial distribution and average size. PMID- 11726026 TI - Benchmarking safety climate in hazardous environments: a longitudinal, interorganizational approach. AB - Safety climate is an important element of organizational reliability. This study applied benchmarking strategies for monitoring safety climate across nine North Sea oil and gas installations that were surveyed in consecutive years. Examination of absolute changes in safety climate complemented the benchmarking approach. Discriminant function analyses (DFA) identified the elements of safety climate predictive of self-reported accidents; correlational analyses were applied to the scale scores and accident proportions across the year period. Absolute improvements were substantial, with safety climate profiles converging in the second year. Large relative improvements were also observed. DFA highlighted perceived management commitment to safety and willingness to report accidents as significant predictors of personal accident involvement. Changes in perceived management commitment to safety were closely associated with changes in safety behavior. PMID- 11726027 TI - Options in radioactive waste management revisited: a proposed framework for robust decision making. AB - Deregulation, with concurrent pressure on electricity utilities, has fundamentally changed the once-"closed" radioactive waste management system controlled by the so-called "nuclear establishment." Advocacy coalitions may change-who knows in which direction-but policy learning may also take place. This article presents a framework to evaluate the management options for a specified concept of "sustainability." When weighing the different objectives in view of the long-lasting potential danger of radiotoxic substances, the overall goal of a sound waste management system is to demonstrate safety. The first-priority objective of a disposal system, therefore, is its stability so that it can comply with the protection goal, that is, the long-term protection of humans and the environment from ionizing radiation. The complementary objective is flexibility, defined here as intervention potential. Because trade-offs within the "sustainability triangle" of ecology, economy, and society are unavoidable, the concept of "integral robustness"-both technical and societal-is introduced into radioactive waste management. A system is robust if it is not sensitive to significant parameter changes. In the present case, it has to have a conservative, passively stable design with built-in control and intervention mechanisms. With regard to technical implementation, a concept called "monitored long-term geological disposal" is presented. Such an "extended" final disposal concept emphasizes technical robustness, recognizes evaluation demands (for a potential break-off of a project), and enhances process-based transparency. This open approach admittedly sets high challenges with regard to technicalities as well as the institutional setting and the management process. It requires "mutual learning" by and from all stakeholders to achieve a truly sustainable radioactive waste management system. PMID- 11726028 TI - Antigen receptor signaling is subverted by an immunomodulatory product secreted by a filarial nematode. AB - ES-62 is a phosphorylcholine (PC)-containing glycoprotein secreted by the rodent filarial nematode Acanthocheilonema viteae which is able to inhibit antigen receptor-stimulated proliferation of B and T lymphocytes in vitro and in vivo. The active component of ES-62 appears to be PC, as the results obtained with ES 62 are broadly mimicked by PC conjugated to bovine serum albumin or PC alone. Such desensitization of lymphocyte responsiveness appears to reflect an uncoupling of the antigen receptors from key intracellular proliferative signaling events, such as the phosphoinositide-3-kinase, protein kinase C and Ras mitogen-activating protein kinase pathways. ES-62 mediates such immunomodulatory effects at concentrations equivalent to those found for PC-containing molecules in the bloodstream of parasitized humans and, thus, ES-62 provides a model system for dissecting the mechanisms of immune evasion induced by related PC-containing glycoproteins expressed by human filarial nematodes. PMID- 11726029 TI - Enhancing cytotoxic T cell responses with altered-peptide ligands. AB - Interest in class I MHC-mediated immunotherapy is growing rapidly. In order to fight a virus or cancer effectively, a successful immunotherapeutic must activate a large number of specific CD8+ T cells and also generate immunological memory. Attempts to generate immune responses towards tumor- or virus-derived peptides have frequently been frustrated by the nature of the peptide antigen itself. Either the peptide does not bind well to its cognate MHC, or the T cells directed towards it have been functionally inactivated in vivo. Altered-peptide ligands (APL) are an effective way to circumvent these problems. However, generating enhanced binding of altered peptides to class I MHC while still maintaining recognition of the wild-type peptide is not straightforward. Many groups design enhanced binding peptides by substituting the observed anchor residues with those that are most preferred by the class I MHC molecule. For many antigenic peptides, this approach does not work. Furthermore, if a higher affinity peptide is designed, the substitutions may result in reduced recognition by CD8+ T cells. Therefore, the design of APL requires careful testing of each candidate therapeutic in terms of affinity for class I MHC and immunological reactivity. Lastly, immunotherapy using class I MHC must also take into account the large genetic heterogeneity in the population. A therapeutic that is only effective for 5-10% of the population is not as attractive as one that works for over 90% of the population. The use of MHC supertypes (groups of class I MHC allotypes that share similar peptide-binding characteristics) shows great promise in overcoming this problem. PMID- 11726030 TI - Multi-functional roles of Stat3 revealed by conditional gene targeting. AB - Signal transducer and activator of transcription (STAT) is a family of transcription factors composed of seven members. Gene-targeted mice of each STAT family protein displayed defective responses to cytokines, demonstrating an important role in cytokine-mediated biological responses. However, unlike the mice lacking other STAT proteins, Stat3-deficient mice died during their early embryogenesis. Therefore, in an attempt to avoid the lethality and assess the role of Stat3 in cytokine-mediated functions in mouse adult tissues, conditional gene targeting utilizing a Cre-loxP system was achieved. By this method, Stat3 was disrupted in several types of tissue, including T cells, macrophages, skin, and mammary gland. Analyses of these Stat3-mutant mice revealed important roles of Stat3 in biological functions in each tissue. PMID- 11726031 TI - Redox control of cellular function by thioredoxin; a new therapeutic direction in host defence. AB - Compelling evidence has suggested that oxidative stress mediates various cellular responses, and control of reduction/oxidation (redox) is important in maintaining the homeostasis of an organism. The thioredoxin (TRX) system, as well as the glutathione system, is one of the key systems in controlling cellular redox status. TRX is a small ubiquitous protein with the redox-active site sequence Cys-Gly-Pro-Cys-. It has been demonstrated to be a multifunctional protein, which has regulatory roles in cellular signaling and gene transcription in addition to cytoprotective activities through the quenching of reactive oxygen species. Various oxidative stimuli, such as UV irradiation, cytokines and some chemicals, promptly induce the expression of TRX. Overexpression of TRX correlates with a wide variety of oxidative stress conditions and, in some cases, TRX has shown promising effects for clinical use, for instance in the attenuation of tissue injury in ischemia reperfusion models. The modulation of TRX functions in association with other redox-regulatory molecules should give us a new therapeutic strategy in the treatment of oxidative stress-mediated disorders and diseases. PMID- 11726032 TI - Cytokine-based immunotherapy of allergic disease. AB - Human immediate hypersensitivity diseases are strongly associated with an excessive type 2 response to normally innocuous environmental antigens, and are a growing health care concern in developed nations. Commonly prescribed treatments provide effective symptomatic relief, but are unable to consistently ameliorate the underlying cause of allergic disease: the excessive generation of allergen specific Th2 cells. IL-12 and IL-18 are potent inducers of type 1 immunity, and, as such, have been proposed as candidates for treatment of allergic diseases. This review critically assesses the potential of recombinant IL-12 and IL-18 immunotherapy to redirect both de novo and established allergic responses in animal models of human allergic disease to clinically protective immune responses. PMID- 11726033 TI - Evidence for an immunoregulatory role of OX2 with its counter ligand (OX2L) in the regulation of transplant rejection, fetal loss, autoimmunity and tumor growth. AB - Transplantation has emerged as an effective treatment for patients with end-stage organ failure. Current regimens of non-specific immunosuppressive drug treatment, which are needed life-long to prevent graft rejection, have numerous adverse side effects and increase the risk of opportunistic infections and malignancy. A major goal is to develop immunotherapeutic protocols that achieve specific tolerance. Such protocols would decrease and eventually eliminate the reliance on non specific drug therapy. We showed that portal vein delivery of donor antigen prolongs the survival of vascularized and non-vascularized allo- and xeno-grafts, and that increased graft survival is associated with altered cytokine production and augmented expression of the molecule OX2. This review documents further evidence for a more general immunoregulatory role for the interactions of OX2 and its ligand, OX2L. PMID- 11726034 TI - The levels of IL-1beta, IL-4 and IL-6 in the serum and the liver tissue of chronic HCV-infected patients. AB - The pro-inflammatory interleukines play a major role in the progress of chronic hepatitis C. Among the patients with chronic hepatitis C virus (HCV) infection, the morphology of the liver was assessed and the levels of serum and liver-tissue IL-1beta, IL-4 and IL-6 were determined. The levels of the cytokines were related to the liver-tissue changes. RNA-HCV was measured by the RT-PCR method. Cytokine levels of the serum and liver tissue were measured by the Quantikine High Sensitivity test. The levels of serum IL-1beta, IL-4 and IL-6 (0.221, 0.104 and 1.393 pg/ml) in all HCV patients were higher in comparison with healthy adults (0.188, 0.025 and 0.600 pg/ml). The levels of liver tissue IL-1beta, IL-4 and IL 6 (4291.3, p<0.05; 1624.6, p<0.05; 1158.7 pg/g protein) in all HCV patients were higher compared with patients with liver cirrhosis without HBV or HCV infection (2319.9, 553.6 and 756.2 pg/g protein). Patients with HCV infection demonstrated a significant correlation between serum and liver-tissue levels of IL-1beta (Pearson: 0.61, p<0.05) and IL-4 (Pearson: 0.51). The level of serum IL-6 in patients with moderate chronic active hepatitis was higher when compared with patients with mild chronic persistent hepatitis. Among the patients with mild chronic persistent hepatitis, the levels of liver-tissue IL-6 were higher compared with those with moderate chronic active hepatitis. There was no correlation between histology changes and the levels of serum and liver-tissue IL 1beta and IL-4. PMID- 11726035 TI - Soluble selectin profiles associated with severe trauma. AB - Severe trauma acts as a trigger for the complex cascade of postinjury events leading to the release of different mediators and the development of generalized inflammation. Selectins are a family of adhesion proteins that are responsible for the adherence of polymorphonuclear neutrophils to the endothelium. This interaction plays an important role in the development of severe complications after multiple trauma. The aim of the present study is to follow the sequential alterations in circulating selectin levels after severe injury and to evaluate the clinical significance of these mediators in monitoring prognosis and outcome. Thirty four severely traumatized patients were entered into the study. Serum sE selectin, plasma sP-selectin and sL-selectin concentrations were measured and an APACHE II score was calculated on admission to the intensive care unit and during the subsequent 5 days. The patients were divided into survivors and nonsurvivors. Initial soluble P- and E-selectin concentrations were significantly elevated in all trauma patients. The highest values of these adhesion molecules were measured in all the observed days in patients with poor prognosis and outcome. In survivors we found a systematic decrease in the sP-selectin concentrations. On admission, the sL-selectin concentrations in all trauma patients were decreased. There were stable, very low values in nonsurvivors and a slow increase in circulating L-selectin in patients who survived. The pattern of soluble selectins in patients with severe trauma is characterized by increased levels of P- and E selectin and a decreased concentration of L-selectin. These findings suggest a widespread microvascular endothelial activation on injury in the early posttraumatic period, which may be associated with increased neutrophil endothelial adhesion, neutrophil extravasation and migration. We suppose that these parameters of endothelial cell activation/injury may be useful as another early prognostic factor in severe trauma. PMID- 11726036 TI - Lactoferrin regulates the immune responses in post-surgical patients. AB - The effect of oral administration of lactoferrin (LF) was studied to determine if it could modify post-surgical immune response. The action of LF was evaluated in 18 LF-treated patients vs 28 placebo counterparts. Patients (women and men, mean age 50 years) were given daily oral doses (20 mg each) of LF for 5 consecutive days prior to thyroid surgery. The following immune response parameters were determined in blood samples taken from the patients day before, day after, and 5 7 days following surgery: cell morphology, the proliferative response of peripheral blood mononuclear cells to phytohemagglutinin, and the spontaneous and lipopolysaccharide (LPS)-induced production of tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6). As a consequence of the thyroid surgery, the total leukocyte count increased on the post-operative day by about 50% in all patients and the percentage of lymphocytes fell by 26 and 35% in the control vs LF-treated group. The content of neutrophils, on the other hand, elevated on day 1 post-operation by 51 and 68%, respectively. The percent of neutrophil precursors was markedly higher in LF-treated patients, particularly on the day before and the day after surgery (4.1 and 4.8 vs 2.5 and 3.7%, respectively). The post-surgical values were, however, comparable in both groups for neutrophils. The proliferative response of lymphocytes showed a slight decrease in the control group and an increase in the LF-treated patients on day 5 post-operation (20% over control group). LPS-induced TNF-alpha production was higher in LF-treated patients both one day before and one day following surgery (28 and 24%, respectively). LPS-induced IL-6 production was comparable in both placebo and LF treated patients before surgery, however, on day 1 and 5 following surgery, the production of IL-6 was higher in LF-treated patients by 65 and 27%, respectively. Taken together, the data presented in this study revealed an increased immune responsiveness in all patients treated with LF and subjected to thyroid surgery. This suggests that treatment with LF could constitute an effective protective measure against post-surgical complications. PMID- 11726037 TI - Human population dynamics. AB - Time-series analysis of parish register series can be used to study human population dynamics at three different levels: (i) The metapopulation of preindustrial rural England. A short wavelength, exogenous oscillation in the burials series of 404 parishes can be detected which, it is suggested, was driven by a cycle of malnutrition associated with wheat prices. (ii) Individual populations, where long-term endogenous oscillations in baptisms and burials of wavelength 30-32 years or 43-44 years can be detected. Their characteristics and causes are explored and elucidated by matrix modelling. (iii) The separate neonatal, post-neonatal, child and adult mortalities in an individual population each show an exogenous short wavelength oscillation and a model is presented to show how these cycles were driven by an oscillation in grain prices and how they interacted. Together, they formed the feedback in a saturated, density-dependent population which was fundamental in controlling the characteristics of the longer wavelength endogenous oscillations in the population dynamics described above. PMID- 11726038 TI - Age and sex variations in adiposity and central fat distribution among elderly Bengalee Hindus of Calcutta, India. AB - BACKGROUND: Although a sizeable portion of India's population (13%, over 110 million) is elderly (aged > 55 years) very little information exists on their levels of adiposity and central body fat distribution. AIM: The present study seeks to investigate age and sex variations in adiposity and central fat distribution among urban elderly Bengalee men and women. SUBJECTS AND METHODS: A cross-sectional study of 410 (210 men and 200 women) elderly (> 55 years) urban Bengalee Hindu individuals resident in Calcutta, India, was undertaken utilizing various measures of adiposity and central fat distribution. RESULTS: There existed significant sex differences in various anthropometric variables and indices. Age had significant negative association with most variables and indices in both sexes. In general, the associations were much stronger in men. Regression analysis demonstrated that age had significant negative effect on height, sitting height (SH), weight, body mass index (BMI), minimum waist (MWC), maximum hip (MHC) and mid upper arm (MUAC) circumferences and triceps skinfold (TSF), in both sexes. Age also had significant negative impact on conicity index (CI) in men. CONCLUSIONS: The present investigation revealed that there is a significant inverse age trend in adiposity among urban elderly Bengalee Hindus. Moreover, there existed sex differences in the effect of age on various anthropometric measures. PMID- 11726039 TI - Relationship between morphologic somatotypes and standing posture equilibrium. AB - Previous studies have identified height and weight as important factors affecting quiet standing stability but studies have not addressed body morphology as a global factor. Using anthropometric measurements, the morphologic somatotypes were defined in terms of body composition and structure. The aim of this study was to test the hypothesis that morphologic somatotypes were related to standing posture equilibrium in able-bodied girls. A total of 43 able-bodied girls having a mean age of 13.8 +/- 2.2 years participated in this study. Somatotype measurements were taken to determine their endomorphic, mesomorphic or ectomorphic components. Then, subjects were asked to stand still on a force platform for 64 s with their eyes opened, feet about 23 cm apart and arms aligned with the trunk. Afterwards, subjects were grouped based on the highest value of their somatotype component. There was no statistical difference in age, height and weight among the groups. The surface area of an ellipse delineated by the displacement of the centre of pressure (COP) was statistically larger (236.9 +/- 134.3 mm2) for the ectomorphs than for the endomorphs 137.7 +/- 71.4 mm2). The minor axis was longer (8.1 +/- 2.9 mm) for the ectomorphs than for the endomorphs (5.7 +/- 2.2 mm). The decrease in standing posture stability of the ectomorphic group was attributed to a relatively low muscle component, a high height weight ratio and an elevated position of the body centre of mass in this population of girls. Somatotypes should be considered when assessing standing posture in both able-bodied subjects and patients. PMID- 11726040 TI - Smoking behaviour and biological maturation in males and females: a 20-year longitudinal study. Analysis of data from the Amsterdam Growth and Health Longitudinal Study. AB - PRIMARY OBJECTIVE: (1) Describe the longitudinal smoking behaviour of boys and girls during adolescence in relation to calendar age, skeletal age, years from peak height velocity (PHV) and years from menarche (in girls). (2) and (3) Investigate the timing of biological maturation (early or late maturation) in relation to smoking behaviour in adolescence and in adulthood (i.e. calendar age 32/33). HPOTHESIS: We hypothesized skeletal age, years from PHV and years from menarche to be better predictors of smoking than calendar age. RESEARCH DESIGN: This study is part of the Amsterdam Growth and Health Longitudinal Study (AGAHLS) that was started in 1977 with 619 pupils from two secondary schools (mean age 13.0 SD 0.6). METHODS AND PROCEDURES: Smoking behaviour was assessed four times between 1977 and 1980 and once in 1996/1997. Calendar age and skeletal age were measured annually whereas height and menarche were measured every 4 months. Maturation rate (skeletal age minus calendar age), age at PHV and age at menarche were used to estimate timing of biological maturation. Generalized Estimating Equation (GEE) analysis was used to study maturation rate in relation to smoking during adolescence, whereas logistic regression analyses were used to study mean maturation rate, years from PHV and years from menarche in relation to smoking in adulthood. OUTCOME AND RESULTS: Skeletal age, years from PHV and years from menarche are no better predictors of smoking during adolescence than calendar age. The prevalence of smoking rises gradually with the increase in all four estimates of biological maturation. Timing of biological maturation was positively related to smoking but only at calendar age 13 (OR 3.34, CI 1.58, 7.07). None of the three measures to estimate timing of biological maturation was significantly related to smoking status in adulthood. PMID- 11726041 TI - Growth of upper- and lower-body segments in Patamona and Wapishana Amerindian children (cross-sectional data). AB - PRIMARY OBJECTIVE: To describe the upper- and lower-body growth of two populations of lowland Amazonian Amerindian children. RESEARCH DESIGN AND METHODS: The stature and sitting height of a convenience sample of 659 Patamona and 543 Wapishana children of known ages between 0 and 16 years of age was collected using standard techniques. Subischial leg length was calculated as stature less sitting height. The raw data were compared with reference data of British children, and body segment z-score values were calculated. RESULTS: The data suggest that Amerindian children were significantly shorter than their British contemporaries, and that considerable faltering in total stature occurred by the third year of life. However, there were contrasting patterns of faltering in the upper and lower body which may well reflect differences in the tempo of growth of body segments at different ages. The two study populations differed significantly in their total stature and this was shown to be due to significant differences in leg length but not in sitting height. CONCLUSIONS: The adverse effects on childhood growth of poor environmental conditions are well known, and it is argued that the variation in body size found between the populations in this study were the results of marked differences in the quality of their living conditions. This paper provides baseline data on the growth of Amerindian populations that are currently facing unprecedented changes in their economic and environmental circumstances. PMID- 11726043 TI - Hip simulator wear testing according to the newly introduced standard ISO 14242. AB - Ultra-high molecular weight polyethylene (UHMWPE) acetabular cups were tested against alumina-ceramic femoral heads using a new type of hip joint simulator according to ISO/FDIS 14242-1. Bovine serum as well as newborn calf serum were used as test fluids. Total polyethylene wear was determined by weight loss of the cups. In addition. wear depth and its distribution were recorded by means of a coordinate measurement system. Wear particle analysis and inspection of the worn polyethylene surfaces using light and scanning electron microscopy (SEM) were performed to analyse damage and identify the acting wear mechanisms. The total wear rate was determined to be 22.07 +/- 1.75 mg/10(6) cycles for the bovine serum group and 26.57 + 3.55 mg/10(6) cycles for the calf serum group. Unexpectedly, the formation of two wear vectors corresponding to recent clinical findings was detected. Retrieved polyethylene wear debris was comparable in size and shape with clinical findings. The test method described by ISO/FDIS 14242-1 produced reliable and reproducible wear data using UHMWPE acetabular cups articulating against alumina-ceramic heads. In the authors' opinion, the lubricant composition should be described in more detail, since the protein and additive content seem to have a high impact on the wear results. It needs to be emphasized that the findings of this study cannot be regarded as a general validation of hip wear tests according to ISO/FDIS 14242-1 but are limited to the material combinations investigated herein. Further testing of other clinically relevant materials and interlaboratory ring tests must follow. PMID- 11726042 TI - The relationship of soluble ICAM-1, VCAM-1, P-selectin and E-selectin to cardiovascular disease risk factors in healthy men and women. AB - BACKGROUND: Clinical studies have shown that elevated serum concentrations of cell adhesion molecules such as inter-cellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1), E-selectin (ESEL) and P-selectin (PSEL) may be independent risk factors for atherosclerosis and cardiovascular disease (CVD). Less is known of the relationship of these inflammatory markers with established CVD risk factors in healthy individuals, particularly women. OBJECTIVE: The aim of this study was to examine cross-sectional relationships between the concentrations of soluble adhesion molecules (sICAM-1, sVCAM-1, sPSEL and sESEL) and smoking behaviour, body composition, blood pressure, serum lipids and physical activity in a large sample of healthy men and women, with special emphasis on interactions between smoking and other CVD risk factors. SUBJECTS: The analysis included 592 healthy white adults aged 18-82 years. RESULTS: There were no sex differences in the concentrations of sICAM-1, sVCAM-1 and sPSEL, but men had higher sESEL levels than women (p < 0.0001). Male and female smokers had higher sICAM-1 and sESEL levels than non-smokers and soluble cell adhesion molecules (CAMs) were correlated with the pack-years of cigarette smoking (r = 0.3-0.4, p < 0.0001, significant in women only). Significant independent associations were found between soluble CAMs and smoking, waist-hip ratio (WHR), blood pressure, high density lipoprotein cholesterol and total cholesterol. Furthermore, significant interaction effects were found in women, such that the relationship between CAMs and lipid concentrations and WHR were stronger in smokers than non-smokers. In conclusion, the concentration of soluble CAMs, particularly sICAM-1 and sESEL, reflect the level of established CVD risk factors in apparently healthy men and women, adding to the evidence that these factors contribute to CVD through their inflammatory effects on the vascular endothelium. PMID- 11726044 TI - An in vitro wear study of alumina-alumina total hip prostheses. AB - Four 28 mm diameter alumina-alumina hip prostheses were tested in the Mkll Durham hip simulator for 5 x 10(6) cycles using 25 per cent bovine serum as lubricant. Wear of the heads and cups was measured gravimetrically. The mean and standard deviation of the wear rate for the alumina cups was 0.097 +/- 0.039 mm3/10(6) cycles. The femoral heads produced such low wear that it could not be measured by weighing but could be detected byincreased surface roughness measurements. Such low wear rates represent about one-five-hundredthof the wear of ultra-high molecular weight polyethylene (UHMWPE) against ceramic in a similar test and supports work which indicates that fluid film lubrication exists in these joints. PMID- 11726045 TI - The effect of socket design, materials and liner thickness on the wear of the porous coated anatomic total hip replacement. AB - The wear of joint replacement prostheses represents the greatest challenge to their continued development. Parameters such as polyethylene quality, liner thickness and metal backing have all been implicated as potential detractors in the search for the lowest-wearing socket. This study examined the effect of these parameters through an extensive study of the two versions of the porous coated anatomic (PCA) hip prosthesis (one-piece socket and snaplock socket). For the whole cohort the wear rate was found to be 88 (SE 10) mm3/year and the clinical wear factor was 2.00 (SE 0.28) x 10(-6) mm3/N m. When the two socket types were investigated individually, the wear factors found were 2.39 (SE 0.44) x 10( 6)mm3/N m and 0.99 (SE 0.25) x 10(-6) mm3/N m for the one-piece and snaplock, respectively. This illustrates that the metal backing per se does not predispose these sockets to rapid wear. The good wear performance of the snaplock liner may be attributed to the high quality of the ultra-high molecular weight polyethylene (UHMWPE) used and the shorter implantation period compared to that for the one piece design. No correlation was found between the thickness of the liner and the clinical wear factor. Within the range of thicknesses tested here, UHMWPE thickness is not an influential parameter for the hip prosthesis and this is confirmed PMID- 11726046 TI - A CAD-CAM methodology to produce bone-remodelled composite femurs for preclinical investigations. AB - Femoral bone remodelling, following total hip arthroplasty, is a clinically observed phenomenon attributed to the changed stress environment of the postoperative implanted hip. While this process cannot be avoided, there is concern as to its consequences on the long-term survival of hip joint replacements. Previous methods of studying remodelling, such as clinical or animal-based studies, or finite element analyses, have their limitations. The aim of this study is to develop experimental specimens incorporating bone resorption features typical of clinically successful implants. This work describes the use of computer aided design/manufacturing methods (CAD-CAM) to produce these specimens, based on modifying commercially available composite femurs. The procedures are investigated and verified for two different designs of cemented prostheses (Lubinus SPII and Muller Curved). Quantitative clinical data is used to define the remodelled geometry of a CAD model of the femur for each stem design. Composite femur specimens are machined using a three-axis milling machine, where each specimen can be accurately positioned using a custom-designed jig and a digitizer system. The accuracy of the process is assessed by analysing the deviation of the digitized premachined and postmachined surfaces of each specimen in relation to the CAD model. The results demonstrate that the procedure can be used for developing in vitro specimens with bone resorption features. These specimens are proposed as a useful tool for performing preclinical trials, such as load transfer or longevity/stability testing, with the advantage of modelling a long-term clinical situation, rather than solely analysing implanted femurs in an immediate postoperative state. PMID- 11726047 TI - Design revision of a partially cemented hip stem. AB - In a previous preclinical study the prototype version of a partially cemented hip stem, cement-locked uncemented (CLU) prosthesis, showed optimal primary stability and moderate stress shielding. However, numerical analysis suggested that the prototype design would induce relatively high stresses in the cement and a significant relative motion between cement and metal. The present study aimed to verify if these problems could be eliminated once the CLU design is improved. The revised design was analysed using a complete finite element model of an implanted human femur. To further strengthen the predictions of the finite element analysis, the cement damage induced by a severe load history was assessed experimentally in synthetic femurs implanted with the improved CLU stem or with a clinically successful fully cemented stem. The modifications made to the CLU stem design did not reduce its good primary stability but decreased the metal-cement relative micromotion. The same load induced stresses in the cement mantle of the improved CLU stem that were significantly lower than those predicted for the prototype design. Although the presence of modelling artefacts produced a highly localized stress peak of 13 MPa. 99 per cent of the cement volume was subjected to a principal tensile stress lower then 4 MPa. These levels of stress compare favourably with the tensile fatigue limit of the acrylic cement used in this study (9.7 MPa). The experimental results further supported these findings. The cemented stem showed a number of cracks per volume unit approximately ten times higher than the partially cemented stem under investigation. PMID- 11726048 TI - A novel three-dimensional tissue equivalent model to study the combined effects of cyclic mechanical strain and wear particles on the osteolytic potential of primary human macrophages in vitro. AB - The effects of cyclic mechanical strain and challenge with physiologically relevant doses of submicrometre-size polyethylene (PE) particles on the osteolytic potential of primary human mononuclear phagocytes were investigated. Cells were seeded into a three-dimensional tissue matrix and co-cultured with particles (mean size 0.21 microm) at particle volume to cell number ratios of 7.5, 15, 30 and 100 microm3/cell. Matrices were then either cultured statically or subjected to 20 per cent cyclic compressional strain in the 'ComCell' for 16 h prior to the assessment of cell viability and quantification of the pro inflammatory cytokine tumour necrosis factor alpha (TNFalpha). The MTT (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyltetrazdium bromide) assay was shown to be too insensitive to detect changes in cell viability. However, when quantified by the adenosine triphosphate (ATP) assay, cell viability was demonstrated to be reduced following exposure to cyclic strain. Macrophages cultured in the static three dimensional tissue equivalent model produced very high levels of TNFalpha in response to submicrometre PE particles at a ratio of 100 microm3/cell. Cyclic strain in the absence of particles gave only a small increase in TNFa production. However, the combined effects of strain and particle stimulation at a ratio of 30 microm3/cell resulted in the secretion of significantly more TNFalpha than was produced by macrophages subjected to strain alone, or the cells-only control. This synergy between cyclic strain and PE particle stimulation was only evident when the volume of particles was reduced below the volume that maximally stimulated cells. These results suggest that while cyclic strain may not be the primary factor responsible for macrophage activation and periprosthetic osteolysis, at low particle load, it may contribute significantly to the osteolytic potential of macrophages in vitro or in vivo. PMID- 11726049 TI - Finite element biphasic indentation of cartilage: a comparison of experimental indenter and physiological contact geometries. AB - In experimental cartilage indentation studies, the indenter is typically a plane ended or hemispherically ended cylinder and can be either porous or non-porous. Joints such as the hip and knee, however, have much higher radii of curvature than those used in experimental indentation testing. In order to interpret the results from such testing in a physiological context it is therefore useful to explore the effect of contact geometry on the pore pressure and strain distribution generated in the cartilage layer. Articular cartilage can be described as a saturated porous medium, and can be considered a biphasic material consisting of a porous, permeable solid matrix, and an interstitial fluid phase. This behaviour has been predicted in this study using the ABAQUS soils consolidation procedure. Four contact geometries were modelled: two typical experimental configurations (5 mm radii cylindrical indenter and hemispherical indenters) and two effective radii representative of the hip and knee (20 and 100 mm). A 10 per cent deformation, or a load of 0.9 kN, was applied over a ramp time of 2 s, which was then maintained for a further 100 s. The porous indenter generated less pore pressure compared with the equivalent non-porous indenter and produced higher values of compressive strain in the solid matrix. The predictions made using the cylindrical indenters, porous and non-porous, were dominated by the concentrations at the edge of the indenter and overestimated the peak compressive strain in the tissue by a factor of 21 and a factor of 14 respectively when compared with the hip model. The hemispherical indenter predicted peak strains in similar positions to those predicted using physiological radii, however, the magnitude was overestimated by a factor of 2.3 when compared with the knee and by 5.7 when compared with the hip. The pore pressure throughout the cartilage layer reduced significantly as the radius of the indenter was increased. PMID- 11726050 TI - Disc hydration measured by magnetic resonance imaging in relation to its compressive stiffness in rat models. AB - The in vivo effects of body dehydration on the water content and compressive stiffness of rat intervertebral disc (IVD) and vertebral body were investigated. Thirty-one rats were divided into three dietary groups: water restricted ( WR), pair fed (PF) and ad libitum (AL). The in vivo water content of L4-5 IVD was measured using magnetic resonance imaging (MRI). Water contents of L4-5 IVD were also determined using the wet/dry weight method after killing. Compressive stiffness of the L5 vertebral body and the L3-4 motion segment were determined. Results showed that the water content of L4-5 IVD was significantly less in the WR group than in the PF and AL groups as measured by the wet/dry weight method; likewise, MRI water content was also less in the WR group. The material stiffness of the IVD was significantly greater in the WR than in the PF and AL groups. However, there was no significant difference in material stiffness of the L5 vertebral body for the WR group in comparison with the PF and AL groups. It was also found that the stiffness of the IVD was significantly lower than that of the vertebral body. This study suggests that the dehydrated matrix of the IVD may be responsible for a stiffer disc that may lead to a less flexible spine. PMID- 11726051 TI - Scaling laws for wall shear stress through stenoses under steady and pulsatile flow conditions. AB - Most patients with atherosclerosis exhibit isolated stenoses of one or more epicardial coronary arteries. The wall shear stresses produced in high-grade stenosis are important in the understanding of atheromatous plaque rupture and thrombosis. This study is designed to establish a method which can be used to scale the different wall shear stresses obtained under different flow conditions to be normalized and subsequently collapsed on to a single general curve. The simulations include both steady and pulsatile flow. The reduced area percentages of the stenoses studied are 50, 75 and 90 per cent. Scaling laws for steady and pulsatile flow conditions are proposed and presented. It can be found from the results that the scaling analysis for pulsatile flow conditions is more complicated than for steady flow conditions and is restricted to, and only valid at, certain time intervals. PMID- 11726053 TI - I'll be alright, Jack. PMID- 11726052 TI - Chest and abdominal surface motion measurement for continuous monitoring of respiratory function. AB - The spirometric devices currently available are not well suited for continuous or ambulatory monitoring of subjects. The proposed solution details a method of assessing lung function that avoids the need for flow measurement at the mouth and provides more detailed information of the dynamics of chest motion while allowing the subject to move freely. The solution to this problem comprises a distributed sensor system that allows regional motions during breathing manoeuvres to be investigated as well as a measurement of tidal respiratory volume. Preliminary studies demonstrate performance benefits of an array of sensors over two-compartment respiratory measurements. PMID- 11726054 TI - Minimal data sets. PMID- 11726055 TI - Teething trouble. PMID- 11726056 TI - Hazards and effects of mobile phones. PMID- 11726057 TI - Assessment of CAL. PMID- 11726058 TI - Unusual presentation of multiple sclerosis. PMID- 11726060 TI - More to chew over. PMID- 11726059 TI - Goltz syndrome. PMID- 11726061 TI - Founders' and Benefactors' lecture 2001. Preventing the preventable--the enigma of dental caries. PMID- 11726062 TI - Good occlusal practice in removable prosthodontics. AB - The loss of teeth may result in patients experiencing problems of a functional, aesthetic and psychological nature. This section addresses the very important subject of occlusal considerations for partial and complete dentures. The occlusion is particularly important given the bearing that occlusal factors have, especially on edentulous patients. PMID- 11726063 TI - Primary care clinicians' knowledge of oral cancer: a study of dentists and doctors in the North East of England. AB - OBJECTIVE: To compare the knowledge of oral cancer and related issues of general dental and general medical practitioners (GDPs and GMPs). DESIGN: A questionnaire was sent to 420 primary care clinicians, half to dentists and half to doctors. Responses to the questionnaire were analysed using odds ratios (OR) and 95% confidence intervals (CI). RESULTS: The response rate was 68.1% for GDPs and 71.9% for GMPs. Dental practitioners were significantly more likely to have diagnosed cases of oral cancer than medical practitioners (OR = 2.68, 95% CI 1.6,4.4). Important differences arose between the groups in terms of risk factor knowledge and clinical examination. CONCLUSION: This study demonstrates that whilst the knowledge level of both groups is generally good, there are some discrepancies. These relate mainly to risk factor knowledge and clinical examination techniques. PMID- 11726064 TI - Salivary cortisol determinations and self-rating scales in the assessment of stress in patients undergoing the extraction of wisdom teeth. AB - OBJECTIVE: To investigate salivary cortisol levels in patients undergoing third molar removal under local and general anaesthesia. DESIGN: Random sampling of two independent groups. METHODS: Samples of saliva were collected from patients prior to and after surgery for the removal of impacted wisdom teeth. The samples were analysed to assess cortisol levels. Patients were also asked to complete a Hospital Anxiety and Depression questionnaire on the day of surgery. RESULTS: Patients receiving treatment under local anaesthesia showed lower levels of stress response than those having treatment under general anaesthesia on the actual day of surgery did (Fig. 1). No other significant differences were noted. CONCLUSION: Local anaesthesia is the preferred mode of pain control for patients who express no strong preference for type of anaesthetic. PMID- 11726066 TI - Anecdotally evidence based. PMID- 11726065 TI - Competency-based education in a clinical course in conservative dentistry. AB - The conceptual difference between a competency-based education and an education based upon a conventional dental curriculum is, perhaps, the starting point for the development of new curricula. The two systems are not, in themselves, exclusive. There is common ground to be found, and the concept of combining instruction with competency-based learning experiences is emerging in recent publications. This paper describes a competency-based approach applied to a clinical course in conservative dentistry. The learning outcomes of the fourth year dental students in the new course were assessed using methods that included continuous clinical assessments, student presentations and peer-group reflective evaluations, patient management reviews, a clinical progress examination, the range and amount of work completed, and a written examination and viva voce. Different weightings were given to various elements of the assessment. A formal student assessment of the course rated it as being satisfactory. PMID- 11726067 TI - Sex with children is abuse: Comment on Rind, Tromovitch, and Bauserman (1998). AB - B. Rind, P. Tromovitch, and R. Bauserman (1998) reported a meta-analysis of the relation between sexual abuse in childhood and adolescence and psychological functioning among college students. Several aspects of their work have proven to be highly controversial, including their assertion that the relation between child sexual abuse and adjustment is quite small and their questioning of whether child sexual abuse should be labeled abuse in scientific inquiry. In this commentary, the authors summarize the controversy that has ensued, place it in a historical context, discuss the limitations of B. Rind et al.'s findings, and critique the manner in which those findings are presented. The authors also argue for the appropriateness of the term abuse and for scientific terminology that reflects rather than contradicts consensual public morality. PMID- 11726068 TI - The effects of child sexual abuse: Comment on Rind, Tromovitch, and Bauserman (1998). AB - B. Rind, P. Tromovitch, and R. Bauserman (1998) examined the long-term effects of childhood sexual abuse (CSA) by meta-analyzing studies of college students. The authors reported that effects "were neither pervasive nor typically intense" and that "men reacted much less negatively than women" (p. 22) and recommended value neutral reconceptualization of the CSA construct. The current analysis revealed numerous problems in that study that minimized CSA-adjustment relations, including use of a healthy sample, an inclusive definition of CSA, failure to correct for statistical attenuation, and misreporting of original data. Rind et al.'s study's main conclusions were not supported by the original data. As such, attempts to use their study to argue that an individual has not been harmed by sexual abuse constitute a serious misapplication of its findings. PMID- 11726069 TI - The validity and appropriateness of methods, analyses, and conclusions in Rind et al. (1998): A rebuttal of victimological critique from Ondersma et al. (2001) and Dallam et al. (2001). AB - The authors respond to 2 victimological critiques of their 1998 meta-analysis on child sexual abuse (CSA). S. J. Dallam et al. (2001) claimed that B. Rind, P. Tromovitch, and R. Bauserman (1998) committed numerous methodological and statistical errors, and often miscoded and misinterpreted data. The authors show all these claims to be invalid. To the contrary, they demonstrate frequent bias in Dallam et al.'s criticisms. S. J. Ondersma et al. (2001) claimed that Rind et al.'s study is part of a backlash against psychotherapists, that its suggestions regarding CSA definitions were extrascientific, and that the moral standard is needed to understand CSA scientifically. The authors show their suggestions to have been scientific and argue that it is Ondersma et al.'s issue-framing and moral standard that are extrascientific. This reply supports the original methods, analyses, recommendations, and conclusions of Rind et al. PMID- 11726070 TI - Empirical evaluation and justification of methodologies in psychological science. AB - The purpose of this article is to describe a relatively new movement in the history and philosophy of science, naturalism, a form of pragmatism emphasizing that methodological principles are empirical statements. Thus, methodological principles must be evaluated and justified on the same basis as other empirical statements. On this view, methodological statements may be less secure than the specific scientific theories to which they give rise. The authors examined the feasibility of a naturalistic approach to methodology using logical and historical analysis and by contrasting theories that predict new facts versus theories that explain already known facts. They provide examples of how differences over methodological issues in psychology and in science generally may be resolved using a naturalistic, or empirical, approach. PMID- 11726071 TI - Development of gender differences in depression: an elaborated cognitive vulnerability-transactional stress theory. AB - Descriptive epidemiological studies are reviewed, showing that the female preponderance in depression begins to emerge around age 13. A developmentally sensitive, elaborated cognitive vulnerability-transactional stress model of depression is proposed to explain the "big fact" of the emergence of the gender difference in depression. The elaborated causal chain posits that negative events contribute to initial elevations of general negative affect. Generic cognitive vulnerability factors then moderate the likelihood that the initial negative affect will progress to full-blown depression. Increases in depression can lead transactionally to more self-generated dependent negative life events and thus begin the causal chain again. Evidence is reviewed providing preliminary support for the model as an explanation for the development of the gender difference in depression during adolescence. PMID- 11726072 TI - The effects of stereotype activation on behavior: a review of possible mechanisms. AB - Considerable recent research has examined the effects that activated stereotypes have on behavior. Research on both self-stereotype activation and other stereotype activation has tended to show that people behave in ways consistent with the stereotype (e.g., walking more slowly if the elderly stereotype is activated). Interestingly, however, the dominant account for the behavioral effects of self-stereotype activation involves a hot motivational factor (i.e., stereotype threat), whereas the dominant account for the behavioral effects of other-stereotype activation focuses on a rather cold cognitive explanation (i.e., ideomotor processes). The current review compares and contrasts the behavioral research on self- and other-stereotype activation and concludes that both motivational and cognitive explanations might account for effects in each domain. PMID- 11726073 TI - Necessity and natural categories. AB - Our knowledge of natural categories includes beliefs not only about what is true of them but also about what would be true if the categories had properties other than (or in addition to) their actual ones. Evidence about these beliefs comes from three lines of research: experiments on category-based induction, on hypothetical transformations of category members, and on definitions of kind terms. The 1st part of this article examines results and theories arising from each of these research streams. The 2nd part considers possible unified theories for this domain, including theories based on ideals and norms. It also contrasts 2 broad frameworks for modal category information: one focusing on beliefs about intrinsic or essential properties, the other focusing on interacting causal relations. PMID- 11726074 TI - Associative learning of likes and dislikes: a review of 25 years of research on human evaluative conditioning. AB - Evaluative conditioning refers to changes in the liking of a stimulus that are due to the fact that the stimulus has been paired with other, positive or negative stimuli. Although evaluative conditioning appears to be subjected to certain boundary conditions, significant evaluative conditioning effects have been obtained using a large variety of stimuli and procedures. Some data suggest that evaluative conditioning can occur under conditions that do not support other forms of Pavlovian conditioning, and several models have been proposed to account for these differences. In the present article, the authors summarize the available literature, draw conclusions where possible, and provide suggestions for future research. PMID- 11726075 TI - Difficult differential diagnoses in IBD: ileitis and indeterminate colitis. AB - When considered in the appropriate clinical scenarios, the diagnoses of Crohn's disease or ulcerative colitis are usually straightforward. Most cases can be definitively diagnosed by the typical subacute or chronic history of symptoms, and radiographic, endoscopic, and histologic confirmation in the presence of negative stool studies; newer serologic assays are now available and are of value if the diagnosis remains uncertain. In this paper, we review distinguishing features in the diagnosis of ileitis and the distinction to be made in conclusively diagnosing ulcerative vs. Crohn's colitis. PMID- 11726076 TI - Self-care for the inflammatory bowel disease patient: what can the professional recommend? AB - The management of patients with inflammatory bowel disease (IBD) continues to present a challenge for gastroenterologists. IBD constitutes a significant burden for the affected individual because of both intestinal and systemic manifestations. The lack of a specific cause and a definitive cure for this condition, as well as the potential for associated malignancy, leave patients unsettled and fearful of the future. Many patients and their families are significantly impaired by the limitations that IBD places on their lives. These limitations are frequently overlooked, and extend well beyond the boundaries of traditional medical practice. Patients often seek to become active participants in the management of their disease. An approach that includes discussion and education on lifestyle issues affecting IBD patients should be part of our routine practice. This article reviews lifestyle and self-care factors relevant to the IBD patient, and what the gastroenterologist can recommend based on current evidence. PMID- 11726077 TI - Managing the glucocorticoid dependent inflammatory bowel disease patient. AB - At this time, standard therapy for treatment of inflammatory bowel disease includes the use of glucocorticoids for moderate to severe Crohn's disease, severe ulcerative colitis, and moderate ulcerative colitis failing mesalamine. Although the majority of patients will have clinical improvement with glucocorticoids, a substantial minority of patients will later flare with attempts to withdraw therapy. Given the numerous potential side effects associated with glucocorticoids, every effort should be made to switch these patients to a less toxic medication. Historically, the most reliable agents have been azathioprine or 6-mercaptopurine. Infliximab is a relatively new medication but would be expected to be beneficial for weaning glucocorticoids for Crohn's disease patients. Methotrexate is another alternative for Crohn's disease. Budesonide and CDP571 are still in developmental phases but likely will be helpful in managing this patient population. Mesalamine, cyclosporine, mycofenalate mofetil, and thalidomide have less data available to support their use but may be helpful for some patients. 6-Thioguanine may be an alternative to patients who do not tolerate 6-mercaptopurine or azathioprine. PMID- 11726078 TI - Management of inflammatory bowel disease during pregnancy and nursing. AB - The peak age of onset for inflammatory bowel disease (IBD) coincides with the peak age for conception and pregnancy, and gastroenterologists will frequently be called on to treat pregnant IBD patients. The greatest threat to a normal conception and pregnancy is active disease, not active medicine. The majority of IBD medications are safe in pregnancy and nursing and should be used as needed. When in remission, ulcerative colitis and Crohn's disease usually do not affect fertility. Fertility may be impaired, however, by pelvic adhesions and scarring from old operations or disease. Pregnant IBD patients should be followed in a facility where diagnostic tests, such as sigmoidoscopy and ultrasound, and surgery can be performed if necessary. PMID- 11726079 TI - Growth failure in the child with inflammatory bowel disease. AB - Once considered rare in pediatric practice, chronic inflammatory bowel disease (IBD) is now being recognized with increasing frequency in children of all ages. In IBD, growth failure may be the only clinical presentation; it is imperative to perform a detailed history and physical examination to search for other systemic and gastrointestinal manifestations of the disease. IBD can have a significant impact on linear growth, weight gain, and bone mineralization, and can cause delays in the onset of puberty. Delays in growth and sexual development can be early indicators of disease activity, and assessment of growth and development should be performed frequently. Nutritional therapy is important not only to correct undernutrition, but also as therapy for IBD. Delayed puberty can have a significant impact on the self-esteem of the adolescent patient and diminish final adult height. Loss of bone mineral density is especially significant during a period in which the majority of bone accretion is expected to occur. These issues present unique problems to the gastroenterologist caring for a child or adolescent with IBD and require specific types of monitoring and interventions. PMID- 11726080 TI - Medical management of ulcerative proctitis, proctosigmoiditis, and left-sided colitis. AB - Ulcerative colitis distal to the splenic flexure includes disease confined to the rectum (proctitis), rectosigmoid (proctosigmoiditis or distal colitis), or extending to the descending colon or splenic flexure (left-sided colitis). These subtypes represent up to 60% to 80% of newly presenting cases of ulcerative colitis. Although these conditions are defined by the extent of colon that is affected, they also share the characteristic of being amenable to topical therapy. In general, the course of disease is milder and symptoms are less severe than in patients with more extensive colonic involvement. Nonetheless, symptoms may significantly impair patients' health-related quality of life. Treatment options include the oral and/or rectal 5-aminosalicylate (5-ASA) preparations. Rectal therapy delivering higher concentrations of active medication (5-ASA or glucocorticoids) directly to the inflamed mucosa while minimizing systemic absorption provides a highly effective and safe treatment. Oral glucocorticoids are indicated in patients who are resistant to or intolerant of 5-ASA therapy. Immunomodulators have an important role in individuals with glucocorticoid dependent or glucocorticoid refractory disease. This article reviews the clinical diagnosis and current medical management of ulcerative proctitis, proctosigmoiditis, and left-sided ulcerative colitis, including patients resistant to conventional medical therapy. PMID- 11726081 TI - Management of bone loss in inflammatory bowel disease. AB - Decreased bone mineral density, osteoporosis, is a common disorder in the United States and elsewhere in the world. This disorder is estimated to account for more than 1.5 million bone fractures each year and is estimated to cost over 13 billion dollars annually. Inflammatory Bowel Disease is a relatively common disorder with estimates suggesting that approximately one million Americans are afflicted by these conditions. Osteoporosis is a common extraintestinal complication of inflammatory bowel disease. The importance of the many possible mechanisms contributing to the loss of bone mass in IBD is uncertain. Several of these specific factors, including the use of corticosteroids, calcium deficiency, vitamin D deficiency, chronically active inflammatory bowel disease, and malnutrition. Assessment of the bone mineral density is an important exercise since new therapeutic options exist and new options are available to combat and even prevent this costly complication. PMID- 11726082 TI - Treatment of acute liver failure. PMID- 11726083 TI - Attention deficits in minimal hepatic encephalopathy. AB - Minimal hepatic encephalopathy (HE) is characterized by a decrease of psychomotor speed, and deficits in visual perception, visuo-spatial orientation, and visuo constructive abilities. Attention deficits have also been proposed to be part of the syndrome. Several attempts were made in the past to elaborate suitable psychometric means for the assessment of minimal HE. However, there is still no "gold standard" for the diagnosis of minimal HE. We recently evaluated the so called "PSE-Test" for the assessment of minimal HE, a test battery which does not include a test predominantly aimed at the assessment of attention. We therefore presented a battery of attention tests in addition to the PSE-Test to a group of cirrhotics without clinical signs of HE compared to a healthy control group matched for age and education to determine whether the addition of special attention tests would increase the diagnostic sensitivity of the PSE-Test. It was shown that the patients with a pathological PSE-Test result differed significantly from controls in all attention tests applied, while the patients with normal PSE-Test results achieved attention test results similar to that of the controls. Thus, the PSE-Test results represent attention deficits as well as deficits in motor skills, visuo-spatial orientation, and visual construction. PMID- 11726084 TI - Review: surgical shunts and encephalopathy. AB - Historically, surgical shunts have played an important role in the treatment of patients with portal hypertension associated with ascites and/or variceal esophageal bleeding. Today, in the era of liver transplantation most patients with end-stage liver disease and concomitant portal hypertension and associated problems are best treated by liver grafting. The successful introduction of transjugular intrahepatic portosystemic shunting (TIPS), performed by radiologists and gastroenterologists, provides a very effective alternative to surgical shunt procedures. One advantage of TIPS is that this procedure does not interfere with subsequent liver grafting. Today, surgical shunts have clearly lost ground to the less invasive TIPS procedure. Surgical shunts still maintain a role: as a salvage procedure in selected cases and in emergency situations. Surgical shunts are associated with a high rate of encephalopathy. In most cases selective surgical shunts should be preferred to nonselective surgical shunts. The role of partial surgical shunts versus selective surgical shunts remains to be determined. Hepatic encephalopathy is a common complication of all shunt procedures and is dependent on the shunt volume. Liver grafting is able to reverse encephalopathy because of a shunting procedure. In our institution, we prefer TIPS over surgical shunts as a bridging procedure before liver transplantation. PMID- 11726085 TI - Movement dysfunction and hepatic encephalopathy. AB - Hepatic encephalopathy is characterized by a variety of neurological symptoms. The occurrence of movement disorders is exceptional and is usually part of a clinical syndrome called acquired hepatocerebral degeneration, which is a subtype of chronic recurrent hepatic encephalopathy. The clinical picture is usually progressive and pathologic findings include regional astroglial and neuronal abnormalities found predominantly in cortex and basal ganglia. As for hepatic encephalopathy in general, the pathophysiology of this disorder is unknown but hyperammonemia and/or brain manganese overload may play a role. Medical treatment is often disappointing but in selected cases liver transplantation may be curative. PMID- 11726086 TI - Neuropsychiatric complications of liver transplantation. AB - Liver transplantation (LT) is the best treatment for end-stage liver diseases but it entails a high incidence of neuropsychiatric complications. These may be related to the operation or occur postoperatively, usually within the first month. The occurrence of neurological problems after LT increases the risk of mortality. The etiology of such complications is various, often multifactorial, immunosuppression being one of the most important causes. Immunosuppressive drugs may cause a wide spectrum of neuropsychiatric complications--mainly affecting the CNS--ranging from mild to severe disorders. A survey of the most frequent disorders is presented. In the management of liver-transplanted patients, the awareness of potential neurological and psychiatric problems is crucial for patients' survival, since it assists clinicians in prevention, prompt diagnosis, and treatment. PMID- 11726087 TI - Quality of life in cirrhotics with minimal hepatic encephalopathy. AB - Minimal hepatic encephalopathy (HE) is a major cause of premature retirement in cirrhotics. The decision on the earning capability of a patient is usually based on clinical judgement, considering the patient's complaints and clinical findings such as nervousness and depression. In a comprehensive psychometric study we were able to show that cirrhotic patients, who are considered to be unable to earn their living, differ significantly from those who are working, in tests evaluating psychomotor function and in personality and subjective well-being scores representing nervousness, aggressiveness, depression. The latter scores are considered to represent the individual discrepancies between professional demands and cerebral performance. Since minimal HE affects psychomotor function but not verbal abilities this discrepancy exists predominantly in "blue collar workers." In accordance with this 60% of "blue collar" (in contrast to 20% of "white collar") workers of our patient group were considered unfit for work. Working ability is an essential element of quality of life in Western societies. Thus, an impairment of working capability is of major impact on quality of life in cirrhotics. PMID- 11726088 TI - Visual event-related potentials in cirrhotic patients without overt encephalopathy: the effects of flumazenil. AB - The P300 complex was derived from the electroencephalogram (EEG) as subjects mentally counted infrequent large checkerboard visual stimuli, presented randomly among frequent small checkerboard stimuli. Use of low contrast (10%) stimuli and four midline scalp electrodes, facilitated separation of cognitive and sensory components and enabled the P300 complex to be resolved into three distinct components--N200, P3a, and P3b. In 20 healthy adult subjects normative data were established and the P3a and P3b components were shown to depend on cognitive function. In 19 age-matched cirrhotic patients without overt hepatic encephalopathy (HE) the EEG and visual evoked potentials (VEPs) were normal, but latencies of P3a and/or P3b were prolonged in 9. Prolonged latencies were not associated with an abnormal number connection test. Ten additional age-matched cirrhotic patients without overt HE, who were alcohol, drug, and caffeine free, were randomized to receive flumazenil (1 mg) and placebo intravenously, double blind. After flumazenil or placebo, latencies of P3a and P3b and psychometric test results did not change significantly. These findings suggest that in cirrhotic patients without overt HE (i) impaired cognitive sensory function may occur in the absence of abnormalities of a standard psychometric test, the EEG, or VEPs, and (ii) increased latencies of P3a and P3b may constitute a component of subclinical HE, which is not mediated by increased brain levels of central benzodiazepine receptor agonist ligands. PMID- 11726089 TI - Neurotransmitter dysfunction in hepatic encephalopathy: new approaches and new findings. AB - Hepatic Encephalopathy (HE) is a serious neuropsychiatric condition of both acute and chronic liver failure. Acute liver failure is characterized by rapid evolution of HE and by brain edema. Portal-Systemic encephalopathy (PSE) is particularly prevalent following treatment of portal hypertension or ascites by the TIPS procedure. Available evidence currently suggests that neurotransmission changes rather than brain energy failure are the primary cause of HE. Recent studies both in autopsied brain tissue from HE patients as well as in experimental animal models of HE reveal that liver failure results in altered expression of several genes coding for proteins having key roles in the control of neuronal excitability. Such alterations include decreased expression of the glutamate transporter GLT-1, and increased expression of monoamine oxidase (MAO-A isoform), the "peripheral-type" benzodiazepine receptor (PTBR) as well as constitutive neuronal nitric oxide synthase (nNOS). Such changes result in altered protein expression and in increased extracellular brain glutamate, increased degradation of monoamine neurotransmitters, increased synthesis of neurosteroids with inhibitory properties, and increased production of nitric oxide (respectively) in brain in chronic liver failure. In the case of GLT-1, PTBR, and nNOS, alterations in expression result from exposure to ammonia and/or manganese, two neurotoxic agents shown previously to be increased in brain in liver failure. PMID- 11726090 TI - Cerebral energy metabolism in hepatic encephalopathy and hyperammonemia. AB - Hepatic encephalopathy (HE) is an important cause of morbidity and mortality in patients with severe liver disease. Although the molecular basis for the neurological disorder in HE remains elusive, elevated ammonia and its chief metabolite glutamine are believed to be important factors responsible for altered cerebral functions, including multiple neurotransmitter system(s) failure, altered bioenergetics, and more recently oxidative stress. Accumulated evidence suggests that direct interference of ammonia at several points in cerebral energy metabolism, including glycolysis, TCA cycle, and the electron transport chain, could lead to energy depletion. Additionally, recent studies from our laboratory have invoked the possibility that ammonia and glutamine may induce the mitochondrial permeability transition in astrocytes, a process capable of causing mitochondrial dysfunction. Altered mitochondrial metabolism appears to be an important mechanism responsible for the cerebral abnormalities associated with HE and other hyperammonemic states. PMID- 11726091 TI - The central opioid system in liver disease and its complications. AB - The best known example of an interaction between the liver and the brain is hepatic encephalopathy. In the 90s a central nervous system origin of the pruritus of cholestatic liver disease and more recently of fatigue of liver disease has been suggested. Hence, three important manifestations of liver disease may be of central origin. Evidence is accumulating that the central opioid system is involved in the development of these manifestations. This short review summarizes current knowledge on the role of the opioid system in development of these liver disease manifestations. PMID- 11726092 TI - Pathophysiology of brain edema in fulminant hepatic failure, revisited. AB - We have proposed a combined osmolar-hemodynamic disturbance to explain the presence of brain edema in fulminant hepatic failure, a major cause of death in this disorder. The concept of an osmotic disturbance in the brain, emphasizing the presence of astrocyte swelling and low-grade cerebral edema, has been expanded to the entire spectrum of liver disease. The mechanism of cerebral hyperemia in patients with FHF and brain swelling has been studied in experimental models linking hyperammonemia and glutamine generation in astrocytes to the development of this hemodynamic alteration. Measures to control cerebral hyperemia, such as mild hypothermia, are effective in preventing the development of brain edema in experimental models as well as intracranial hypertension in human disease. PMID- 11726094 TI - Fine needle aspiration cytology diagnosis of cutaneous leishmaniasis in a 6-month old child: a case report. AB - BACKGROUND: Skin biopsy and scrape smear examination are the two most commonly employed investigatory techniques in the diagnosis of cutaneous leishmaniasis. Although cases Leishmania lymphadenitis are reliably diagnosed with fine needle aspiration (FNA) cytology, it has not attained popularity in the diagnosis of cutaneous leishmaniasis, and only a few reports are available. CASE: A 6-month old Kuwaiti child presented with a skin lesion on her left forearm of five months' duration. Both scrape smears and FNA were performed from the lesion. FNA cytology smears showed a rich population of inflammatory cells predominating in lymphocytes and histiocytes and epithelioid cell granulomas. The amastigote forms of Leishmania were noted on the smears. The scrape smears were nondiagnostic. CONCLUSION: FNA cytology can be reliably used in the diagnosis of cutaneous leishmaniasis, especially in dry lesions, where scrape smears are likely to be nondiagnostic. PMID- 11726093 TI - Mild hypothermia prevents cerebral edema and CSF lactate accumulation in acute liver failure. AB - Evidence from both clinical and experimental studies demonstrates that mild hypothermia prevents encephalopathy and brain edema in acute liver failure (ALF). As part of a series of studies to elucidate the mechanism(s) involved in this protective effect, groups of rats with ALF resulting from hepatic devascularization were maintained at either 37 degrees C (normothermic) or 35 C (hypothermic), and neurological status was monitored in relation to cerebrospinal fluid (CSF) concentrations of ammonia and lactate. CSF was removed via implanted cisterna magna catheters. Mild hypothermia resulted in a delay in onset of encephalopathy and prevention of brain edema, CSF concentrations of ammonia and lactate were concomitantly decreased. Blood ammonia concentrations, on the other hand, were not affected by hypothermia in ALF rats. These findings suggest that brain edema and encephalopathy in ALF are the consequence of ammonia-induced impairment of brain energy metabolism and open the way for magnetic resonance spectroscopic monitoring of cerebral function in ALF. Mild hypothermia could be beneficial in the prevention of severe encephalopathy and brain edema in patients with ALF awaiting liver transplantation. PMID- 11726095 TI - Infarction after fine needle aspiration biopsy of pleomorphic adenoma of the parotid gland. AB - BACKGROUND: Fine needle aspiration (FNA) is a well-established and safe method for the rapid diagnosis of pleomorphic adenoma. A few clinically important complications, including bleeding, infection and inflammatory reactions, result from FNA. In a small number of cases FNA has been followed by varying degrees of necrosis in some organs. In the literature there are a few reports associated with necrosis in a pleomorphic adenoma of the parotid gland following FNA. CASE: A 27-year-old female had a two-year history of a right parotid mass. FNA revealed pleomorphic adenoma. A histologic diagnosis of pleomorphic adenoma of the right parotid with infarction was made. CONCLUSION: Necrosis associated with infarction may cause diagnostic problems. It is not a sufficient sign of malignant transformation. PMID- 11726096 TI - Diagnosis of malignant granular cell tumor by fine needle aspiration cytology:. AB - BACKGROUND: Malignant granular cell tumors (GCT) are the rarest of all sarcomas, and the histologic differentiation from their benign counterpart may be extremely difficult or impossible unless metastatic disease is demonstrated. To our knowledge, this is the first report of a malignant GCT diagnosed by fine needle aspiration (FNA) cytology. CASE: A 70-year-old, Caucasian female presented with a progressively enlarging left supraclavicular mass. FNA of the mass revealed a metastatic tumor cytologically consistent with GCT. With this diagnosis, a search for other metastatic sites was initiated. Computed tomography (CT) scan revealed several tumor nodules in the lungs and liver. CT-guided FNA and tru-cut needle biopsy of a liver mass confirmed the diagnosis of metastatic GCT. In searching for a primary site, the patient revealed a clinical history of having had a tumor removed from her back two months before; it was reported to be an atypical GCT. Comparison of the three tumors revealed similar histologic, cytologic and immunohistochemical features. CONCLUSION: Evidence of mild to moderate cytologic atypia; increased mitotic activity; locally aggressive growth; increased proliferative activity as demonstrated by immunohistochemical evaluation of proliferation markers; and DNA ploidy analysis, as reported for this case, may be helpful in predicting malignant behavior of GCTs. PMID- 11726097 TI - Hepatic metastases from the spindle cell variant of medullary thyroid carcinoma: report of a case with diagnosis by fine needle aspiration biopsy. AB - BACKGROUND: The liver is a common site of neuroendocrine tumors (NTs) metastatic from primaries in the gastrointestinal tract, pancreas, biliary system and lungs. Medullary thyroid carcinoma (MTC) is also a potential source of metastases of NTs. Their metastases to the liver are frequent and can appear several years after the primitive tumor. Although a wide variety of cytomorphologic features are normally exhibited by MTC in smears, a spindle-shaped cell pattern can predominate, complicating the correct interpretation of a metastasis. CASE: A 63 year-old man presented with multiple liver nodules two years after a total thyroidectomy for MTC. Fine needle aspiration biopsy smears of the liver revealed neoplastic cells occurring in loose groupings or lying singly, most of them with a spindle shape and elongated nucleus with the characteristic "salt and pepper" chromatin pattern of a neuroendocrine tumor. Cytoplasmic dendritic processes and intranuclear inclusions were frequently seen. The cytomorphologic features of the tumor were essentially the same as those of the primary MTC. Immunoreactivity for calcitonin confirmed the diagnosis. CONCLUSION: In fine needle aspiration biopsy of liver masses, knowledge of the spindle pattern of the NT is important in order to achieve a correct diagnosis when metastases are the first manifestation of an occult primary tumor. Among neuroendocrine tumors, MTC must be included in the differential diagnosis. PMID- 11726098 TI - Fine needle aspiration cytology of mammary duct ectasia: report of a case with novel cytologic and immunocytochemical findings. AB - BACKGROUND: Mammary duct ectasia (MDE), or periductal mastitis, is a well-defined clinical entity with a characteristic histopathologic appearance on breast biopsy specimens. However, the cytologic features of MDE have been described only recently in the cytopathology literature, and fine needle aspiration (FNA) cytologic findings are based mainly on a small number of case reports in the English-language literature. Therefore, awareness of this entity and recognition of its cytomorphologic features could aid in a more accurate diagnosis. We report the novel cytologic and immunocytochemical findings on a case of MDE that was confirmed by histologic examination on a subsequent biopsy. CASE: We report the findings of breast FNA cytology in a 50-year-old female with a mammographically and clinically suspicious lesion. Cytology displayed a paucicellular aspirate, typified by a few scattered, cohesive clusters of ductal epithelial cells with mild nuclear atypia and distinct, peripherally located myoepithelial cells. Intermingled within the ductal elements were numerous polygonal cells with abundant, finely vacuolated cytoplasm that were immunoreactive for macrophage specific antibody, CD68. The background consisted of a variable number of foam cells and negligible amount of blood. CONCLUSION: The current case of MDE demonstrates cytomorphologic features that may pose diagnostic problems, particularly as a consequence of variable cytologic atypia this entity may present on FNA cytology. This diagnostic difficulty is compounded because of the abnormal mammographic and suspicious clinical findings that may be associated with MDE. CD68 immunoreactivity is a useful ancillary tool to verify the histiocytic, rather than epithelial and potentially neoplastic, nature of multivacuolated cells. To the best of our knowledge, there has been no prior reported case of MDE in the English-language literature studied utilizing CD68 antibody. This case report emphasizes that MDE is a heterogeneous entity with diverse cytomorphologic features. FNA cytology in conjunction with immunocytochemistry might permit accurate classification in the proper clinical setting. PMID- 11726099 TI - Diagnosis of clinically unsuspected extrapulmonary tuberculosis by fine needle aspiration: a case report. AB - BACKGROUND: Mycobacterium tuberculosis (MTb) infection remains the cause of higher morbidity and mortality than any other infectious disease in the world. Intact cellular immunity is necessary to resist the disease, and therefore the AIDS epidemic has greatly contributed to the resurgence of MTb. Depending on the degree of immunosuppression, the presentation of MTb in patients with AIDS can be atypical and difficult to diagnose as compared to the classical presentation of MTb in the nonimmunocompromised population. CASE: A patient who was not known to be HIV positive had a clinical picture of extensive abdominal and pelvic lymphadenopathy without chest radiographic abnormalities. The diagnosis of MTb was made by fine needle aspiration (FNA) of a pelvic lymph node. CONCLUSION: Miliary tuberculosis associated with AIDS may have an unusual clinical presentation and unusual cytologic features on ENA. PMID- 11726100 TI - Cytologic features of desmoplastic infantile ganglioglioma: a report of two cases. AB - BACKGROUND: Desmoplastic infantile ganglioglioma (DIG) is a rare intracranial tumor of infancy, characterized by solid and cystic components, voluminous size and supratentorial location. Although its histologic features have been reported, there has been no cytologic description of the tumor. Cytologic findings on imprint and aspirated material from two cases of histologically verified DIG are discussed. CASES: A 12-month-old male with cutis marmorata telangiectatica congenita and a 44-month-old female with episodes of spasm were referred to our center. Radiologic examination revealed a large, cystic, supratentorial mass in both patients. The mass was surgically removed, and histology revealed prominent desmoplasia with a mixture of astroglial and neuronal cells. Cytologic findings on imprint material and the needle aspirates taken from the cystic area of the tumor revealed a few isolated or sheetlike arrangements of small cells positive for glial fibrillary acidic protein and a few large cells with abundant cytoplasm and prominent nucleoli, positive for neuronal marker. CONCLUSION: Although distinction between ganglioglioma and DIG may be difficult by cytology alone, with the characteristic clinical presentation and radiologic findings, the possibility of DIG should be kept in mind when the specimens contain both astroglial and neuronal elements. PMID- 11726101 TI - Cytomorphologic diagnosis of malignant lymphoma arising in the heart: a case report. AB - BACKGROUND: Primary malignant lymphoma of the heart is extremely rare. Because its clinical signs and symptoms are typically nonspecific, it is often very difficult to detect cardiac involvement while the patient is alive. We describe a case of malignant lymphoma involving predominantly the heart and pericardium and diagnosed by pericardiac effusion cytology antemortem. CASE: An 83-year-old woman presented with dyspnea on exertion. Echocardiography revealed a low-echoic tumor mass close to the right ventricular wall and massive pericardiac effusion. Diagnosis of diffuse large B-cell lymphoma was made by cytomorphologic examination and flow cytometry of the tumor cells obtained from the effusion. Although chemotherapy was instituted immediately, the patient died of progressive heart failure. Diffuse large B-cell lymphoma predominantly involving the intracardiovascular region was confirmed at autopsy. CONCLUSION: From the experience in this case, we conclude that cytopathologic examination of sonographically guided aspiration of the cardiovascular region is very useful for antemortem diagnosis of primary malignant lymphoma of the heart. PMID- 11726102 TI - Peripheral primitive neuroectodermal tumor of the vulva: report of a case with imprint cytology. AB - BACKGROUND: Peripheral primitive neuroectodermal tumor (PNET) of the vulva is an extremely rare disease, and, to our knowledge, only two cases have been previously reported. CASE: A 45-year-old woman presented with a mass in the right labium major. Three years after removal of the tumor, she noticed a new lesion in the same place and underwent a partial vulvectomy. The imprint cytology of the recurrent tumor showed a monomorphic appearance, composed of small round cells with scant cytoplasm against a hemorrhagic background. These tumor cells were loosely connective, but rosettelike structures were observed focally. On pathologic examination, the neoplasm was composed of small round tumor cells showing sinusoidal, diffuse or micropapillary growth. Immunohistochemically, the neoplastic cells stained positively for neuron-specific enolase, vimentin and HBA 71 and negatively for cytokeratin, HBA 45 and muscle-specific actin. The morphologic characteristics of the disease were well expressed in the imprint cytology, and this influenced the selection of immunohistochemical studies. CONCLUSION: Cytologic examination for vulvar tumors, even imprint cytology, can be a useful tool in obtaining an accurate pathologic diagnosis of a rare disease, such as peripheral PNET. PMID- 11726103 TI - Meningioma presenting as a parapharyngeal tumor: report of a case with fine needle aspiration cytology. AB - BACKGROUND: Meningiomas rarely extend out of their intracranial confines through skull foramina to present as cervical tumors, where they would be accessible to fine needle aspiration (FNA) and thereby create difficulties in cytodiagnosis by mimicking other, more commonly aspirated head and neck tumors. CASE: A psammomatous meningioma arising intracranially and extending through the jugular foramen presented as a mass at the angle of the jaw clinically. On FNA cytology the diagnosis was suggested, but the cytomorphologic features overlapped with those of more commonly aspirated head and neck tumors, such as acinic cell carcinoma arising primarily in a salivary gland, metastatic papillary thyroid carcinoma and paraganglioma (glomus jugulare tumor). These possibilities had to be excluded through correlation with radiologic and intraoperative findings, which showed a dural-based tumor extending through the jugular foramen to assume a parapharyngeal location. Histology of the final excision specimen confirmed a psammomatous meningioma. CONCLUSION: The possibility of meningioma should be considered in the cytologic differential diagnosis of parapharyngeal tumors, particularly since its cytomorphologic features may mimic those of some of the more commonly encountered and aspirated head and neck tumors. PMID- 11726104 TI - Cytologic features of a primary myxoid malignant fibrous histiocytoma arising in the uterus: a case report. AB - BACKGROUND: Primary malignant fibrous histiocytoma (MFH) of the uterus is extremely rare. The 10 cases reported in the literature all involved the pleomorphic variant, and to the best of our knowledge, the myxoid variant has not been reported before. We describe the cytologic findings of primary uterine myxoid MFH in relation to the myxoid component, potentially leading to an incorrect diagnosis. CASE: A 68-year-old woman presented with a primary uterine tumor. Endometrial cytology showed numerous loosely arranged, spindle-shaped fibroblastlike cells; atypical histiocytelike cells; and giant cells with a necrotic background. The overall cytologic picture was of a degenerated pleomorphic leiomyosarcoma with an inconclusive diagnosis. A diagnosis of myxoid MFH was established after electron microscopic and immunohistochemical studies of the primary tumor and tumor transplanted, as primary cultured cells, in nude mice. The patient underwent an exploratory laparotomy and died of tumor progression 38 days after the initial consultation, without treatment. CONCLUSION: Because of overlapping cytologic features among uterine sarcomas with myxoid stroma, it is important to recognize the histiocytic lineage of tumor cells by immunohistochemistry and electron microscopy in various presentations of fresh samples. PMID- 11726105 TI - Scrape cytology of meningioangiomatosis: a report of two cases with diagnostic cytologic features. AB - BACKGROUND: Meningioangiomatosis is a rare, probably hamartomatous condition characterized by nonneoplastic intracortical proliferation of meningothelial cells, capillaries and fibroblasts. The lesion may mimic a tumor both clinically and radiologically. We present two cases of the entity, including its cytologic features. CASES: A 71-year-old man presented with an eight-month history of headache, change in vision, loss of depth perception and unsteady gait. Magnetic resonance imaging (MRI) revealed an infiltrative lesion of the left occipital lobe, diffuse in nature. Cytologic smears prepared at the time of intraoperative consultation showed numerous thin-walled capillaries together with bland spindle cells. Occasional large cells with prominent nucleoli were also present. Frozen section confirmed the presence of linear, small capillaries surrounded by fibroblasts and meningothelial cells, consistent with meningioangiomatosis. The second case was a 3-year-old girl who presented with a one-day history of seizure, vomiting, fever and perioral cyanosis. MRI revealed an enhancing lesion in a temporal lobe of the cerebrum. The patient underwent excision of the lesion. Intraoperative cytology showed numerous meningothelial whorls together with neurons and occasional capillaries. Both patients were well, one after three months and the other after six. CONCLUSION: Meningioangiomatosis is a lesion characterized by linear capillaries, meningothelial cells and neurons, some exhibiting nuclear atypia. The combination can lead to an erroneous diagnosis of more common conditions, such as meningioma (with intracortical extension) and anaplastic astrocytoma. Contrary to these more sinister diagnoses, meningioangiomatosis is a completely benign, presumably hamartomatous entity. PMID- 11726106 TI - Aspiration cytology of glomus tumor: a case report. AB - BACKGROUND: Glomus tumor is a relatively uncommon soft tissue tumor that can occur at any age and anatomic site, with a predilection for the subungual region. CASE: A 24-year-old female presented with a gradually enlarging, painful swelling in the subungual region of the right index finger. Fine needle aspiration was performed under a ring block and yielded hemorrhagic material. The smears revealed clusters of uniform, round to oval cells admixed with wisps of magenta intercellular myxoid material and a few spindle-shaped cells crossing clusters of tumor cells. A diagnosis of benign vascular tumor consistent with glomus tumor was given. Subsequent histopathologic examination confirmed the diagnosis and showed alpha-SMA positivity on immunohistochemistry. CONCLUSION: This case report is the second on the cytologic features of glomus tumor in the English-language literature. Although the cytomorphologic features of glomus tumor are quite distinctive, an appropriate clinical history and immunohistochemical stains (e.g., alpha-SMA and vimentin positivity) can further help to ascertain the diagnosis. PMID- 11726107 TI - Fine needle aspiration cytology of eccrine acrospiroma. PMID- 11726108 TI - Fine needle aspiration of pleomorphic liposarcoma of the breast: revised diagnosis. PMID- 11726109 TI - Diagnostic cells in scrape cytology of squamous cell carcinoma. PMID- 11726110 TI - Fine needle aspiration of tumoral calcinosis. PMID- 11726111 TI - Phagocytosis of rod-shaped bacteria and cocci by Trichomonas vaginalis: light microscopic observations. PMID- 11726112 TI - Collagenous spherulosis of the breast. PMID- 11726113 TI - Cytologic features of the tubular variant of breast adenomyoepithelioma. PMID- 11726114 TI - Fine needle aspiration cytology of a meningioma with extracranial extension after irradiation for medulloblastoma. PMID- 11726115 TI - Fine needle aspiration cytology of granular cell tumor in the breast of a male. PMID- 11726116 TI - Conjunctival dysplasia in soldiers exposed to mustard gas during the Iraq-Iran war: scrape cytology. AB - OBJECTIVE: To describe the conjunctival scrape cytology findings in Iraq-Iran war veterans exposed to mustard gas and to detect malignant transformation at an early stage. STUDY DESIGN: Twenty-two male war veterans (29-54 years old) who were exposed to chemical bombing about 14 years earlier, formed the base of this study. All of them had chronic eye problems, such as dryness, conjunctival scarring and decreased visual acuity. Conjunctival scrape cytology was performed for cytologic study. RESULTS: Among 22 cases of conjunctival scarring, 9 were diagnosed as dysplasia. Mild inflammation and squamous metaplasia were also noted. Squamous cell carcinoma was not identified. CONCLUSION: Conjunctival scraping cytology is useful for the diagnosis of dysplasia in soldiers exposed to mustard gas during of the Iraq-Iran war. This method is also effective for follow up study. PMID- 11726117 TI - Expression of bcl-2 protein in bronchoalveolar lavage cell populations from patients with idiopathic pulmonary fibrosis. AB - OBJECTIVE: To investigate changes in the expression of the antiapoptotic protein bcl-2 in bronchoalveolar lavage fluid (BALF) cell populations in patients with idiopathic pulmonary fibrosis (IPF). STUDY DESIGN: Ten patients with IPF underwent fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) in the area of maximal radiographic shadowing (based on high-resolution computed tomography findings). Results were compared with those of 10 normal people in the control group. Cellular bcl-2 expression was identified using an immunoperoxidase staining method. RESULTS: A statistically significant (P < .001) increase in the expression of bcl-2 in BALF neutrophils and eosinophils was observed in patients with IPF as compared with controls. BAL macrophages exhibited only a slight (statistically insignificant) increase in bcl-2 expression in IPF patients. No bcl-2 expression was observed in BAL lymphocytes from IPF patients in contrast to the control group. CONCLUSION: The overexpression of bcl-2 on BALF neutrophils and eosinophils, cells that characterize the special cellular profile of alveolitis in IPF, could be one of the pathophysiologic mechanisms of this disease. PMID- 11726118 TI - Utility of the in situ detection of HPV in Pap smears diagnosed as within normal limits. AB - OBJECTIVE: To determine the clinical significance in normal Pap smears of HPV detection as determined by Hybrid Capture (HC) and in situ hybridization analyses. STUDY DESIGN: We studied 135 consecutive Pap smears as well as 46 other smears from high-risk patients each initially diagnosed as within normal limits. RESULTS: The 135 "normal" Pap smears were rescreened, and 6 (4%) where found to be either ASCUS or SIL. In the remaining 129 cases, HPV DNA was detected in 0% and 9%, respectively, using in situ hybridization and HC I. Upon rescreening the high-risk patients, nine (20%) were reclassified as having SIL/ASCUS; each was in situ hybridization positive, and eight were HC positive; six (67%) of these women developed SIL on follow-up. In the 37 Pap smears in high-risk women still within normal limits after manual rescreening, HPV was detected in 2% by in situ hybridization and 46% by HC; 6% of the HC-positive women developed SIL on follow up. CONCLUSION: In situ hybridization rarely detects HPV in Pap smears diagnosed as within normal limits after manual rescreening. In situ hybridization is very effective in detecting rare, atypical cells in Pap smears diagnosed as within normal limits and, in a high-risk population, is predictive of SIL on clinical follow-up. PMID- 11726119 TI - Transvaginal fine needle aspiration biopsy. AB - OBJECTIVE: To assess the role of transvaginal fine needle aspiration biopsy (FNAB) in the evaluation of palpable gynecologic masses. STUDY DESIGN: Transvaginal FNABs from 1994 to 1999 were identified from the files of Barnes Jewish Hospital. Histologic correlation was obtained using the Pathology Department's computer database. Two pathologists reviewed the pathologic samples. Pertinent clinical information was obtained by reviewing the medical records. RESULTS: Twenty-two transvaginal FNABs from 22 patients were studied. The patients' mean age was 59 years (range, 29-84). Most patients (77%) had a previous history of a gynecologic malignancy, and 73% had a previous total abdominal hysterectomy and bilateral salpingo-oophorectomy. The size of the lesion sampled was provided in 15 cases and ranged from <1 to 5.4 cm in diameter. The location of the mass was reported as follows: vaginal (10 cases), vaginal cuff (5), rectovaginal septum (2), cul-de-sac (1), fornix (1), vaginal apex (1), right side of pelvis (1), and not specified (1). The cytologic diagnoses were: negative for malignancy (10 cases), positive for malignancy (9) and unsatisfactory (3). Most cases (77%) had histologic correlation or clinical follow-up. There was one false negative and no false positive cytologic diagnosis. CONCLUSION: Cytologic interpretation of transvaginal FNAB is an effective toolfor the evaluation of palpable pelvic and vaginal masses. Its specificity and sensitivity are 100% and 88%, respectively. PMID- 11726120 TI - Peritoneal washing cytology on fluid hysteroscopy and after curettage in women with endometrial carcinoma. AB - OBJECTIVE: To assess the influence of fluid hysteroscopy with target biopsy of the endometrium and the influence of added curettage on the results of peritoneal washing cytology (PWC) in endometrial carcinoma. STUDY DESIGN: In 42 women at risk of endometrial carcinoma, we performed fluid hysteroscopy with target biopsy of the endometrium and curettage. Evaluation of PWC of the pouch of Douglas was performed three times during the procedure: prior to hysteroscopy, after fluid hysteroscopy with target biopsy and after curettage. RESULTS: On cytologic slides from peritoneal washings in 11 patients with carcinoma of the endometrium, malignant endometrial cells were found after curettage in 72.7%. There was no statistically significant difference in PWC prior to hysteroscopy (two women, 20%) or after hysteroscopy with target biopsy (three women, 30%). There was a statistically significant difference (.05 level) in positive PWC after hysteroscopy with target biopsy (three women, 33.3%) and after curettage (eight women, 88.9%). CONCLUSION: Slides from carcinoma of the endometrium in PWC do not deteriorate after hysteroscopy with target biopsy of the endometrium, but tumor cells will appear in the pouch of Douglas after curettage. PMID- 11726121 TI - Algae in cytologic smears. AB - OBJECTIVE: To desmonstrate the presence of algae in smears and establish their significance. STUDY DESIGN: Microscopic examination of smears stained by the Papanicolaou technique. RESULTS: We found 12 different species of algae, clustered in three categories: Cyanophita (blue algae), Chrysophyta (yellow algae) and Chlorophyta (green algae). CONCLUSION: Algae in smears are infrequently observed, with few bibliographic references. Their presence is due to intrinsic or extrinsic contamination. They may be confused with other structures, such as fungi, Charcott crystals or adenocarcinoma cells. PMID- 11726122 TI - Aspiration cytology of 147 adnexal cysts with histologic correlation. AB - OBJECTIVE: To assess the usefulness of cytology in the diagnosis of 147 histologically established adnexal cysts. STUDY DESIGN: Retrospective, macro microscopic study based on fluid aspirated from 132 ovarian and 15 extraovarian cysts and projected as a cytohistologic correlation. RESULTS: Typical macroscopic features were identified in 76% of endometriotic cysts, in 53% of mucinous neoplasms and in 67% of dermoid cysts. Cytology helped to identify 67% of nonneoplastic and 56% of neoplastic cysts. The lowest diagnostic sensitivities were observed in functional cysts and benign serous neoplasms (50%), while the highest were shown by endometriotic cysts (76%) and malignant epithelial neoplasms (71%). Inadequate samples were obtained from all types of cysts, even malignant ones (two mucinous cystadenocarcinomas). Diagnostic cytology was useless in extraovarian cysts (33% sensitivity). An adult granulosa cell tumor was erroneously diagnosed as a follicular cyst by cytologic examination. CONCLUSION: Examination of the cyst fluids obtained by aspiration demonstrated low sensitivity, with 43% of inadequate samples obtained from all types of cysts. Malignant cystic neoplasms may be overlooked in inadequate samples. Our study also revealed that specificity in this type of analysis is high in inadequate samples, provided that the technique is carried out correctly. PMID- 11726123 TI - Gynecomastia: cytologic features and diagnostic pitfalls in fine needle aspirates. AB - OBJECTIVE: To illustrate some of the uncommon cytologic findings of gynecomastia, such as apocrine metaplasia, cellular atypia and foamy macrophages, that can be misinterpreted as evidence of malignancy. STUDY DESIGN: The clinical data and fine needle aspiration (FNA) cytologic material from 100 men with the diagnosis of gynecomastia were retrospectively reviewed. The excisional biopsy slides were available for 16 cases. For comparison, FNA smears from five men with breast lesions other than gynecomastia were studied. RESULTS: The patients ranged in age from 23 to 91 years. Cytologic findings were as follows: cohesive sheets of cells containing 20-1,000 cells (98%); scattered, single, bipolar cells (78%); spindle cells (68%); ductal epithelial atypia (26%); apocrine metaplasia (8%); and foamy histiocytes (12%). In nine cases the atypia was marked, and in two of them the possibility of malignancy could not be ruled out. Surgical follow-up on 16 patients, including the cases with marked atypia, showed gynecomastia. In one case, gynecomastia was associated with intraductal papilloma. No correlation between the underlying etiology and atypical cytologic features of gynecomastia was identified. CONCLUSION: Apocrine metaplasia and epithelial atypia are common findings in gynecomastia. Attention to the cell patterns, the presence of sheets of ductal cells and absence of atypical single cells will point to the correct diagnosis. PMID- 11726124 TI - Diagnosing Kikuchi disease on fine needle aspiration biopsy: a retrospective study of 44 cases diagnosed by cytology and 8 by histopathology. AB - OBJECTIVE: To study the accuracy of fine needle aspiration (FNA) diagnosis of Kikuchi lymphadenitis (KL). STUDY DESIGN: Retrospective review of all cases of FNA biopsy of lymph nodes in which KL was diagnosed or suggested. False positive cases were studied. Cases of KL diagnosed by histopathology were examined for the false negative rate of FNA diagnosis. RESULTS: Forty-four cases of KL diagnosed or suggested by FNA were found. Five of eight cases were confirmed on lymph node excision. The false positive rate was 37.5%. One case was nonspecific reactive changes. Two cases were proven to be tuberculous lymphadenitis by culture. Eight cases of KL diagnosed by lymph node excisional biopsy had prior FNA. Four were diagnosed as or suspected to be KL. The false negative rate was 50%. CONCLUSION: The overall accuracy of FNA diagnosis of KL was 56.25%. Detailed study offalse positive cases and knowledge of other conditions suggested that overreliance on certain cytologic features and the morphologic erlap between KL and tuberculous lymphadenitis could have been the reasons for the inaccuracies. PMID- 11726125 TI - Abundance of siderophages in sputum: indicator of an adverse lung reaction to air pollution. AB - OBJECTIVE: To investigate the lung response to traffic-related air pollution by enumerating hemosiderin-laden alveolar macrophages (AM) in sputum. STUDY DESIGN: Sputum samples were collected from 103 urban adult males from Calcutta chronically exposed to automobile exhaust. Forty-nine rural individuals served as controls. AM were identified by nonspecific esterase staining. Perl's Prussian blue technique was employed for the detection of hemosiderin-laden AM (siderophages). RESULTS: The urban group, consisting of 31 traffic officers, 25 automobile service station workers and 47 street hawkers, had seven times more AM in their sputum than did the matched controls. Besides, a remarkable rise (27 fold) in the number of siderophages in sputum was observed in urban individuals. Smoking further elevated the AM count and number of siderophages. CONCLUSION: Abundant siderophages in the urban group may indicate the toxic effect of airborne pollutants on the lung, leading to phagocytosis of destroyed cells, including erythrocytes, and accumulation of iron in AM. Enumeration of siderophages in sputum appears to be a simple, noninvasive, inexpensive cytochemical technique well suited to preliminary assessment of the adverse effects of air pollution on the lungs in large, population-based studies, especially in developing countries. PMID- 11726126 TI - Proliferative activity in invasive breast carcinoma: a comprehensive comparison of MIB-1 immunocytochemical staining in aspiration biopsies to image analytic, flow cytometric and histologic parameters. AB - OBJECTIVE: To use MIB-1 antibody to assess proliferative activity in fine needle aspiration (FNA) samples of invasive breast carcinoma and compare these results to multiple other measures of proliferative activity. STUDY DESIGN: FNA slides from 62 patients with invasive breast carcinoma were subjected to staining with MIB-1. Quantitative MIB-1 values were compared to image analytic proliferative fractions (IPF) obtained from the same FNAs. MIB-1 values were also compared to flow cytometric S-phase fractions (SPF) and S + G2/M-phase fractions (FPF) and to histologic assessment of mitotic count (MC) in resected tumors. RESULTS: MIB-1 values, IPF, SPF, FPF and MC were suitable for evaluation in 55, 53, 50, 50 and 56 cases, respectively. MIB-1 values showed good correlation with IPF in FNAs (correlation coefficient = .57, P <.00001). MIB-1 values also showed correlation with SPF (correlation coefficient =.447, P = .003), FPF (correlation coefficient = .325, P = .023) and MC (correlation coefficient = .402, P = .01) in resected tumors. CONCLUSION: This study supports the use of MIB-1 values obtained from FNA samples for assessment of proliferative activity in invasive breast carcinoma, based on correlation of these values with multiple other parameters of proliferative activity. Assessment of these values can play a role in predicting prognosis and in selecting patients with invasive carcinoma of the breast for preoperative or adjuvant chemotherapy. PMID- 11726127 TI - Fine needle aspiration cytology of invasive micropapillary carcinoma of the breast: review of cases in a three-year period. AB - OBJECTIVE: To describe the fine needle aspiration cytology findings of invasive micropapillary carcinoma and correlate them with the histologic appearance. STUDY DESIGN: We reviewed the cytologic features of three cases of pure invasive micropapillary carcinoma in the files of Pamela Youde Nethersole Eastern Hospital from 1998 through 2000. Immunohistochemical study for epithelial membrane antigen was performed retrospectively on the cell block sections. Ultrastructural examination was also carried out on one of the cases. RESULTS: Two of the tumors were at pathologic stage II, and the remaining case was at stage III. Ipsilateral axillary lymph node metastases with similar morphology were seen in two of them. Cytologically, the smears were of moderate cellularity and composed of three dimensional tumor cell balls, abortive and sometimes branching papillae, angulated tumor cell clusters, morules and occasional acini. Some of the tumor cell balls possessed scalloped borders. Focally, the tumor morules clustered together and were separated from each other by small, slitlike spaces. A small number of isolated malignant cells was also present in the background. The cell block sections showed mainly dispersed acini of tumor cells. The "reverse polarity" highlighted in histologic sections by immunohistochemical study for epithelial membrane antigen was not consistently demonstrated in the cell block material. Ultrastructural examination confirmed the focal presence of surface microvilli on the periphery of the tumor cell morules. CONCLUSION: Invasive micropapillary carcinoma of the breast possesses some subtle but distinctive cytologic features. With the help of cell block morphology and ancillary techniques, the preoperative suspicion of this rare subtype of ductal carcinoma, which carries a high propensity for lymphatic permeation, is possible. PMID- 11726128 TI - Three-dimensional cytomorphology in fine needle aspiration biopsy of medullary thyroid carcinoma. AB - OBJECTIVE: To elucidate three-dimensional (3-D) cytomorphology in fine needle aspiration biopsy (FNAB) of medullary thyroid carcinoma (MTC). STUDY DESIGN: ENAB was performed on tumors from five patients with MTC. The aspirate was stained and observed under a light microscope (LM). The aspirate was also fixed, dehydrated, critical point dried, spattered with gold ions and observed with a scanning electron microscope (SEM). For transmission electron microscopy (TEM), the specimen was fixed, dehydrated, embedded in an Epon mixture, cut with an ultramicrotome, mounted on copper grids, electron doubly stained with uranium acetate and lead citrate, and observed with TEM. Findings under SEM were correlated with those under LM and TEM. RESULTS: Under SEM, 3-D cytomorphology of MTC displayed a disorganized cellular arrangement with indistinct cell borders in three cases. The cell surface was uneven and had granular protrusions that corresponded to secretory granules observed under TEM. In one case with multiple endocrine neoplasia type IIB, there were abundant granules on the cell surface. In one case of sporadic MTC with multinucleated tumor giant cells and small cells, granular protrusions also were noted on the cell surface. CONCLUSION: Granular protrusion was a characteristic finding in FNAB of MTC tinder SEM and might be helpful in the differential diagnosis. PMID- 11726129 TI - Retrospective study of Chlamydia trachomatis using the polymerase chain reaction on archival Papanicolaou-stained cytologic smears. AB - OBJECTIVE: To detect chlamydial DNA on archived Papanicolaou-stained (Pap) smears using the polymerase chain reaction (PCR) technique. STUDY DESIGN: A PCR assay was designed to identify chlamydial DNA using consensus sequences unique to the genus Chlamydia in the 16S rRNA gene. This assay produced a 109 base pair product containing a single Pvu II restriction site. One hundred cervicovaginal Pap smears from a teen clinic population were processed for DNA isolation and PCR. Amplifiable DNA was isolated from 93 of the 100 cases as determined by a human growth hormone gene. These specimens were subjected to chlamydial PCR. RESULTS: PCR analysis of the 93 samples yielded 6 that were positive for the chlamydial 16S rRNA sequence. The six positive chlamydial amplicons were purified and subjected to Pvu II restriction enzyme analysis to validate their identity. The analysis confirmed the identity of the products, as a single Pvu II restriction site resulted in 41 base pair and 68 base pair products, as predicted. CONCLUSION: PCR testing for Chlamydia trachomatis can be performed on DNA isolated from archival Pap smears. Using this methodology, 6.5% of young women in our teen clinic population were positive for chlamydial DNA. PMID- 11726130 TI - Touch imprint cytology in biopsy of the infertile testis. AB - OBJECTIVE: To identify different cell types in the testis by using touch imprint cytology and to compare the cytologic findings to the histopathologic diagnosis in infertile men. STUDY DESIGN: This prospective study used touch imprint preparations and included 20 infertile men. The biopsy material obtained was stained with toluidine blue, May-Grunwald-Giemsa stain and Papanicolaou stain. The cytologic results for oligospermic, normospermic and azospermic men were compared to the specific histopathologic diagnosis. The proportion of spermatogenic versus Sertoli cells was calculated. The scores were compared between three groups based on the results of the histologic biopsy: normal spermatogenesis, hypospermatogenesis and incomplete spermatogenic arrest. RESULTS: The mean ratio of the spermatogenic cells versus Sertoli cells was statistically significantly different in the three groups (P < .01). The mean ratio of spermatogenic cells to Sertoli cells was higher in cases with normal spermatogenesis than in cases with hypospermatogenesis and incomplete spermatogenic arrest, revealing a statistical difference (P<.01). This ratio was not statistically significantly different between the hypospermatogenesis and incomplete spermatogenic arrest groups. CONCLUSION: A cytologic demonstration of germinal cells by using touch imprint preparations may be a guide for histologic diagnosis. PMID- 11726131 TI - Processing liquid-based gynecologic specimens: comparison of the available techniques. AB - OBJECTIVE: To develop a cytopreparation protocol suitable for satisfactory processing by the AutoCyte PREP System with the gynecologic specimens collected in PreservCyt fluid for the ThinPrep machine. STUDY DESIGN: The residue of a number of gynecologic specimens collected in PreservCyt and processed by ThinPrep were processed by AutoCyte PREP. Some modifications were made in the cytopreparatory protocol in order to obtain satisfactory specimens. RESULTS: The ThinPrep and AutoCyte PREP specimens were examined independently. The results were comparable, with a high degree of concordance between the two techniques. CONCLUSION: Gynecologic specimens collected in PreservCyt and following the ThinPrep specimen collection protocol can be processed using the AutoCyte PREP System. Minor protocol modifications provided comparable diagnostic material. Additional studies are needed to explore the feasibility of this approach and fulfill the various U.S. regulatory agency requirements for the liquid-based Pap test. PMID- 11726132 TI - Intraabdominal follicular dendritic cell sarcoma: report of a case with fine needle aspiration findings. AB - BACKGROUND: Follicular dendritic cell sarcoma is a rare tumor derived from dendritic cells of lymphoid follicles. Although it shows characteristic cytologic, histologic and immunohistochemical features, it may be misdiagnosed because of the lack of experience with this entity. There are few reports in the cytologic literature describing this entity. CASE: A left hypochondrial mass was found in an 80-year-old man who had a past history of colonic carcinoma. A computed tomography-guided fine needle aspiration biopsy was performed, and the findings were initially thought to be compatible with metastatic carcinoma. The mass was excised, and a diagnosis of follicular dendritic cell sarcoma was made. CONCLUSION: Follicular dendritic cell sarcoma is characterized by loosely cohesive or syncytial groups of dendritic cells that show oval, vesicular nuclei and ill-defined cytoplasm intimately admixed with small, mature lymphocytes. The diagnosis can be confirmed by positive immunostaining for CD21 and CD35 antigens and negative staining for cytokeratin. Unusual features in our case were the presence of prominent plasma cells and intracytoplasmic, periodic acid-Schiff stain, diastase-positive deposits, with the last not described before. PMID- 11726133 TI - Fever and cancer in perspective. AB - CONTEXT: A relationship between feverish infection and concurrent remission from cancer has been known about for a very long time. However, a systematic investigation of the phenomenon has not yet been made. OBJECTIVE: To bring together the isolated observations about the coincidence of spontaneous remissions with feverish infections and William Coley's seminal work, as a basis for devising an immunological hypothesis about the putative anti-cancer effect of fever. CONCLUSION: Fever induction under medical guidance may be considered as part of a therapy regimen for cancers of mesodermal origin. PMID- 11726134 TI - Transduction of human dendritic cells with a recombinant modified vaccinia Ankara virus encoding MUC1 and IL-2. AB - The epithelial mucin MUC1 is considered an opportune target antigen for cancer immunotherapy, as it is over-expressed and exhibits aberrant glycosylation in malignant cells. Because dendritic cells (DC) are powerful initiators of immune responses, efforts have focused on tumor antigen-bearing DC as potent cancer vaccines. In this study we have characterized the transduction of monocyte derived DC with a highly attenuated vaccinia virus vector [modified vaccinia Ankara (MVA)] encoding human MUC1 and the immunostimulatory cytokine IL-2. Analysis of transduced DC cultures generated from a number of donors revealed MUC1 expression in the range of 27-54% of the cells and a co-regulated secretion of bioactive IL-2. As shown by FACS analysis with MUCI-specific antibodies, the MVA-MUC1/IL-2-transduced DC predominantly expressed the fully processed glycoform of MUC1, typical of that displayed by normal epithelia. Over a 3-day period after transduction, transgene expression declined concurrent with an increase in MVA induced cytopathic effects. The transduced DC stimulated allogeneic lymphocyte proliferation, indicating that DC immunostimulatory function is not impaired by vector transduction. In the presence of IL-2, MVA-transduced DC were able to enhance autologous lymphocyte proliferation. Also, vector expression was analyzed in DC cultures treated with TNF-alpha, a known DC maturation factor. As indicated by the up-regulation of several DC maturation markers, neither virus infection nor transgene expression influenced the maturation capacity of the cells. The MVA MUC1/IL-2 vector effectively transduced both immature and TNF-alpha-matured DC. Overall, our results are encouraging for the clinical application of MVA-MUC1/IL 2-transduced DC. PMID- 11726135 TI - Enhancement of anti-cancer immunity by a lipoteichoic-acid-related molecule isolated from a penicillin-killed group A Streptococcus. AB - We isolated the lipoteichoic-acid-related molecule (OK-PSA) from OK-432, a streptococcal preparation, by affinity chromatography on CNBr-activated Sepharose 4B-bound monoclonal antibody TS-2, which neutralizes the interferon (IFN)-gamma inducing activity of OK-432. We have previously reported that OK-PSA is a potent inducer of Th1-type cytokines in human peripheral blood mononuclear cells in vitro. In this study, we conducted an animal experiment to examine whether OK-PSA exhibits an anti-tumor effect in vivo by acting as a Th1 inducer in syngeneic Meth-A tumor-bearing BALB/c mice, in which the Th2 response is genetically dominant. It was found that OK-PSA induced Th1-type cytokines [IFN-gamma, tumor necrosis factor-alpha, interleukin (IL)-2, IL-12 and IL-18] in BALB/c mice bearing Meth-A tumor and caused a marked anti-tumor effect. Although it was suggested by an in vitro study. using spleen cells derived from the animals, that IL-18 plays the greatest role in the induction of the Th1-dominant state and tumor cell killing induced by OK-PSA, the in vivo experiments demonstrated that both IL-12 and IL-18 are essential in the anti-tumor effect exhibited by OK-PSA. These findings strongly suggest that OK-PSA is a major effector molecule of OK 432 and may be a useful immunotherapeutic agent, as a potent Th1 inducer, for cancer patients with a Th2-dominant state. PMID- 11726136 TI - Standardized generation of fully mature p70 IL-12 secreting monocyte-derived dendritic cells for clinical use. AB - Dendritic cells (DC) have been shown to be efficient antigen-presenting cells (APC) and, as such, could be considered ideal candidates for cancer immunotherapy. Immature DC (iDC) efficiently capture surrounding antigens; however, only mature DC (mDC) prime naive T lymphocytes. Clinical trials using DC based tumor vaccines have achieved encouraging, but limited, success, possibly due to the use of immature or incompletely mature DC. Thus, it was apparent that a method capable of generating large numbers of fully functional iDC, their pulsing with desired form of tumor antigens and the subsequent complete and reproducible maturation of iDC is needed. Therefore, we compared two different methods of producing large numbers of iDC. Both protocols yielded comparable numbers of cells with an iDC phenotype with phagocytic function. We next determined which of the clinically applicable activators could induce the complete and reproducible maturation of DC, in order to define the most suitable combination for future clinical trials. Only a combination of TNFalpha + Poly (I:C), or a previously described cytokine cocktail of TNFalpha + IL-1beta + IL-6 + prostaglandin E2, induced the complete activation of the whole DC population, as assessed by the cell surface expression of CD83 and costimulatory molecules. The matured DC were functionally superior to iDC in their ability to stimulate the proliferation of allogeneic lymphocytes and autologous keyhole limpet hemocyanin (KLH)-specific T lymphocytes. Furthermore, only the combination of TNFalpha + Poly (L:C) activated DC to produce large amounts of biologically active p70 IL-12. Thus DC maturation by TNFalpha + Poly (I:C) could efficiently bias T cell response towards Th1 response. Implementation of our results into clinical protocols used for DC generation could be beneficial for future immunotherapy trials. PMID- 11726137 TI - Target cell induced activation of NK cells in vitro: cytokine production and enhancement of cytotoxic function. AB - This study examines the effect of fixed AK-5 tumour cells on rat NK cells. Co culture of NK cells with fixed tumour cells augmented the cytotoxicity of NK cells against NK-sensitive targets, YAC-1 and AK-5, and induced the secretion of IFN-gamma by NK cells. Antibody against IFN-gamma suppressed the anti-tumour activity of NK cells, whereas the addition of T cells during co-culture enhanced this activity. However, macrophages and B cells had no significant effect when present during co-culture with NK cells. All the inducible cytotoxicity was contained within the NK (CD161+) and NKT (CD3+, CD161+) subsets of lymphocytes. However, in the presence of T cells, the cytolytic potential of NKT cells was higher than that of NK cells alone. The augmentation of cytotoxic activity of NK cells by AK-5 cells in presence of T cells was dependent on IL-2 and IFN-gamma secretion. NK cell activation was blocked by specific antibodies to IL-2 and IFN gamma in the presence of T cells. Interaction between fixed AK-5 cells with NK and T cell populations induced the expression of Fas-L and perforin in NK cells. These data demonstrate that fixed AK-5 cells initiated cytokine synthesis by NK cells, and the enhanced cytotoxic activity in the presence of T cells was induced as a consequence of the products secreted by activated T lymphocytes. The present observations reflect the possible interactions taking place in vivo after the transplantation of AK-5 tumour in animals. They also suggest direct activation of NK cells after their interaction with the tumour cells. PMID- 11726138 TI - A stable single-chain variable fragment expressing transfectoma demonstrates induction of idiotype-specific cytotoxic T-cells during early growth stages of a murine B-lymphoma. AB - The idiotypic determinants associated with the variable regions of antibody molecules are known to function as tumor-associated antigens (TAAs). However, there is no clear-cut evidence documenting their efficacy in inducing TAA specific cytotoxic T-lymphocytes (CTLs). In most previous studies, idiopeptides were implicated in elicitation of TAA-specific CD4+ T-cells. Using a murine B cell lymphoma, 2C3, we earlier demonstrated induction of splenic CD4+ and CD8+ T lymphocytes directed to idiotypic Ig of the tumor. In the present study, we provide more direct evidence of the existence of Id-specific CTLs in the spleens of 2C3 bearing BALB/c mice using an scFv-transfectoma, P815A4, as a target. While both P815A4 and 2C3 cells were equally susceptible to cytolysis by the effector cells, lysis was evident only during early tumor progression. Moribund animals at the late stage of tumor growth failed to demonstrate any significant cytotoxic immune response against either tumor. Antibodies to MHC class I alleles Kd, Dd, Ld, beta2m and CD8 molecules all inhibited cytotoxicity. The CTL population from early tumor-bearers recognized 2C3 tumor in the context of all major H-2d alleles; however, in case of P815A4 cells, it was restricted to Kd and Dd alleles only. Based on these antibody inhibition studies, it appears that the idiopeptides generated in both tumors are in some way different, yet they were recognized equally by CTLs not only from the tumor-bearers but also by CTLs from 2C3-hyperimmune mice. It appears that scFv-containing transfectomas expressing antibody variable region epitopes would be useful for both elucidating CTL defined idiopeptides and monitoring TAA-specific CTL response in tumor-bearing animals. PMID- 11726139 TI - "Differentiation between types and strains of clostridium botulinum by riboprinting," A comment on: J. Food Prot. 63(10):1347-1352 (2000). PMID- 11726140 TI - Use of steam condensing at subatmospheric pressures to reduce Escherichia coli O157:H7 numbers on bovine hide. AB - This study used a laboratory-scale apparatus to apply subatmospheric steam to bovine hide pieces inoculated with Escherichia coli O157:H7 in maximum recovery diluent (MRD) and in high-liquid content and low-liquid content fecal suspensions (HLC fecal and LLC fecal, respectively). The survival of the organism in fecal clods, which were stored for 24 days in a desiccated state, was assessed. Inoculated fecal clods were also treated with subatmospheric steam. Steam treatment at 80 +/- 2 degrees C for 20 s reduced E. coli O157:H7 concentrations on hide inoculated to initial concentrations of approximately 7 log10 CFU/g by 5.46 (MRD inoculum), 4.17 (HLC fecal inoculum), and 5.99 (LLC fecal inoculum) log10 CFU/g. The reductions achieved in samples inoculated with LLC feces were larger than in samples inoculated with HLC feces (P < 0.05). Treatment at 80 +/- 2 degrees C for 10 s resulted in significantly smaller reductions (P < 0.05) on hide pieces of 2.54 (MRD), 1.94 (HLC fecal), and 2.15 (LLC fecal) log10 CFU/g. There were no significant differences among the reductions observed in all inoculum types in samples treated for 10 s. E. coli O157:H7 inoculated in fecal clods to 7.78 log10 CFU/g and stored at 4 or 15 degrees C survived for at least 24 days. Steam treatment (20 s) of 3-day-old clods reduced surviving E. coli O157:H7 numbers from 4.20 log10 CFU/g to below the limit of detection of the assay used (1.20 log10 CFU/g). This study shows that steam condensing at or below 80 +/- 2 degrees C can reduce E. coli O157:H7 when present on bovine hide, reducing the risk of cross contamination to the carcass during slaughter and dressing. PMID- 11726141 TI - Effect of acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue and effect of modified atmosphere packaging on survival of Escherichia coli O157:H7 on red meat. AB - The present study examined the effect of pH-independent acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue. A varied level of acid resistance was observed among the 14 strains tested. Eight strains were categorized as acid resistant, four strains as acid sensitive, and two strains demonstrated acid-inducible acid resistance. The survival of an acid-resistant (II/45/4) and acid-sensitive (IX/8/16) E. coli O157:H7 strain on chilled beef tissue treated with 1 and 2% buffered lactic acid, sterile water, or no treatment (control) was followed. A gradual reduction of E. coli O157:H7 was noticed during the 10 days of storage at 4 degrees C for each of the treatments. Decontamination with 1 and 2% buffered lactic acid did not appreciably affect the pathogen. Differences in the pH-independent acid resistance of the strains had no effect on the efficacy of decontamination. The effect of modified atmosphere packaging (MAP) on survival of E. coli O157:H7 in red meat was also studied. MAP (40% CO2/60% N2) or vacuum did not significantly influence survival of E. coli O157:H7 on inoculated sliced beef (retail cuts) meat compared to packing in air. The relative small outgrowth of lactic acid bacteria during storage under vacuum for 28 days did not affect survival of E. coli O157:H7. Neither lactic acid decontamination nor vacuum or MAP packaging could enhance reduction of E. coli O157:H7 on beef, thus underlining the need for preventive measures to control the public health risk of E. coli O157:H7. PMID- 11726142 TI - Inhibitory activity of Bifidobacterium longum HY8001 against Vero cytotoxin of Escherichia coli O157:H7. AB - Vero cytotoxin (VT)-producing Escherichia coli (VTEC), such as E. coli O157:H7, are emerging foodborne pathogens worldwide. VTs are associated with hemorrhagic colitis and hemolytic uremic syndrome in humans. Attachment of the B subunit of VTs to its receptor, globotriaosylceramide (Gb3), at gut epithelium is the primary step and, consequently, the A subunit of VTs inhibits protein synthesis in the target cell. Proinflammatory cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, up-regulate Gb3 expression, increase sensitivity to VTs, and enhance VT action in developing disease. Currently, there is a growing interest in probiotics, given the increasing occurrence of antibiotic-resistant bacteria. In particular, much work on bifidobacteria among probiotics, regarded as microorganisms targeted for technological and therapeutic applications, has been performed. In Korea, the neutralizing effect of the culture supernatant of Bifidobacterium longum HY8001, Korean isolate, against the VTs from E. coli O157:H7 was found. Therefore, this study focused on the raveling of the inhibitory effect of B. longum HY8001 against VTs, through the interference B subunit of VTs and Gb3 interaction. Mice were inoculated intragastrically with B. longum HY8001 culture supernatant before and after challenge with E. coli O157:H7. Control mice were inoculated intragastrically only with E. coli O157:H7. Cytokine, TNF-alpha, and IL-1beta levels in sera and expression of their mRNA were decreased, and expression of Gb3 in renal tubular epithelial cells was reduced in mice treated with B. longum HY8001 culture supernatant. In competitive enzyme-linked immunosorbent assays (ELISAs), the culture supernatant of B. longum HY8001 primarily binds VTs to interfere the VTs with Gb3 interaction. These results suggest that soluble substance(s) in B. longum HY8001 culture supernatant may have inhibitory activity on the expression of Gb3, VT-Gb3 interaction, or both. Further study should be done to elucidate the property of soluble substances in B. longum HY8001 culture supernatant. PMID- 11726143 TI - Acid tolerance and survival of Escherichia coli O157:H7 inoculated in fruit pulps stored under refrigeration. AB - The adaptation of Escherichia coli O157:H7 cells to acid conditions has been reported. This study showed the behavior of three strains of E. coli O157:H7 in two different physiological stages (acid shocked and control cells), inoculated in five fruit pulps stored at 4 degrees C for up to 30 days. The three strains of E. coli O157:H7 inoculated in grape pulp could be recovered up to day 30. E. coli O157:H7 strains survived for 4 days in all fruit pulps tested with different pH values (2.51 to 3.26), with the exception of acid-shocked cells of E. coli O157:H7 strain 933, which did not survive in "caja" (Spodias lutea L.) pulp. The results clearly indicated that acid resistance can persist for long periods during storage at 4 degrees C. The protection conferred by acid adaptation suggests that acid-resistant organisms will be better equipped to outlast these acid challenges. The survival of E. coli O157:H7 in fruit pulps with a low pH, stored under refrigeration, is of extreme importance due to the high virulence of this microorganism. The viability of microorganisms was dependent on the viscosity index of fruit pulps. Less viscous pulps supported the bacteria survival longer than more viscous fruit pulps. PMID- 11726144 TI - Validation of apple cider pasteurization treatments against Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes. AB - Time and temperature pasteurization conditions common in the Wisconsin cider industry were validated using a six-strain cocktail of Escherichia coli O157:H7 and acid-adapted E. coli O157:H7 in pH- and degrees Brix-adjusted apple cider. Strains employed were linked to outbreaks (ATCC 43894 and 43895, C7927, and USDA FSIS-380-94) or strains engineered to contain the gene for green fluorescent protein (pGFP ATCC 43894 and pGFP ATCC 43889) for differential enumeration. Survival of Salmonella spp. (CDC 0778. CDC F2833, and CDC H0662) and Listeria monocytogenes (H0222, F8027, and F8369) was also evaluated. Inoculated cider of pH 3.3 or 4.1 and 11 or 14 degrees Brix was heated under conditions ranging from 60 degrees C for 14 s to 71.1 degrees C for 14 s. A 5-log reduction of nonadapted and acid-adapted E. coli O157:H7 was obtained at 68.1 degrees C for 14 s. Lower temperatures, or less time at 68.1 degrees C, did not ensure a 5-log reduction in E. coli O157:H7. A 5-log reduction was obtained at 65.6 degrees C for 14 s for Salmonella spp. L. monocytogenes survived 68.1 degrees C for 14 s, but survivors died in cider within 24 h at 4 degrees C. Laboratory results were validated with a surrogate E coli using a bench-top plate heat-exchange pasteurizer. Results were further validated using fresh unpasteurized commercial ciders. Consumer acceptance of cider pasteurized at 68.1 degrees C for 14 s (Wisconsin recommendations) and at 71.1 degrees C for 6 s (New York recommendations) was not significantly different. Hence, we conclude that 68.1 degrees C for 14 s is a validated treatment for ensuring adequate destruction of E. coli O157:H7, Salmonella spp., and L. monocytogenes in apple cider. PMID- 11726145 TI - Sources and movement of Salmonella through integrated poultry operations: a multistate epidemiological investigation. AB - The prevalence of Salmonella from numerous sources in 32 integrated broiler operations of high- and low-performing broiler houses was characterized from four states across four seasons. Previous studies of Salmonella in broilers have been limited in scope, offering only a snapshot of pathogen prevalence as seen on a small number of individual farms. Twenty-six different sample types were collected from the hatchery to the end of processing, and Salmonella was found in all sample types. A total of 10,740 samples were analyzed for Salmonella, and 973 (9.1%) of these samples, including 49 of 798 (6.1%) carcass rinse samples, were Salmonella positive. Hatchery transport pads (389 of 765, 50.8%), flies (28 of 150, 18.7%), drag swabs (57 of 402, 14.2%), and boot swabs (20 of 167, 12%) were samples from which Salmonella was most frequently isolated. Thirty-six different serotypes were identified, and the most frequently encountered serotypes were Salmonella Senftenberg, Salmonella Thompson, and Salmonella Montevideo. Determining critical contaminating sources and following the movement of Salmonella through integrated poultry operations will help researchers and the industry develop practical intervention strategies. PMID- 11726146 TI - Colonization of broiler chicks by Salmonella typhimurium definitive phage type 104. AB - The prevalence of an antibiotic-resistant strain of Salmonella Typhimurium definitive phage type 104 (DT104) has increased dramatically in recent years resulting in increased morbidity and mortality in both animals and humans. Colonization and shedding of Salmonella Typhimurium DT104 was studied in broiler chickens in two trials. In trial 1, 180 day-of-hatch chicks (n = 60 per group, n = 30 per replicate) were challenged with 10(6) CFU DT104 (wild-type isolate from poultry) or were commingled with a seeder chick challenged with 10(6) CFU DT104. In trial 2, 360 day-of-hatch chicks (n = 120 per treatment, n = 30 per rep) were divided into three groups. Chicks in the susceptible group were commingled with two seeder chicks that were orally challenged with 10(7) CFU/bird of a pan sensitive strain of Salmonella Typhimurium DT104. Chicks in the resistant group were commingled with two seeder chicks that were orally challenged with 10(7) CFU/bird DT104 used in trial 1. For both trials, a control group was not exposed to DT104, composite fecal samples were evaluated twice weekly for levels of Salmonella shedding and 20 chicks per group were necropsied weekly and their cecal contents were cultured. At hatch all groups were colonized with naturally occurring Salmonella Senftenberg and Salmonella Mbandaka (trial 1) or Salmonella Senftenberg and Salmonella Ohio (trial 2) prior to exposure to DT104. Throughout the study, the level of Salmonella spp. shedding in feces (trial 1 means 3.1, 2.9, and 3.0 log10 CFU per g feces for challenged, seeder, and control groups, respectively) or ceca (trial 2 means 2.9. 2.9. and 2.5 log10 CFU per g ceca for resistant, susceptible, and control groups, respectively) did not differ among groups. In trial 1, colonization of DT104 remained constant at higher levels in the challenged group (mean 87%, P < 0.01), increased over time in the seeder group (10 to 50%, P < 0.02) and was not recovered from the control chicks. Salmonella Mbandaka colonization remained steady within each group with challenge and seeder groups maintaining higher levels of colonization than the control group. Salmonella Senftenberg colonization levels tended to decline (P = .058) over time in the challenged group (20 to 0%) and significantly decreased (P < 0.01) over time for both the seeder (80 to 0%) and control chicks (85 to 10%). In trial 2, the percentage of chicks colonized with susceptible DT104 declined (r = 0.90, P < 0.05) over the course of the trial from 45 to 0%, while recovery of the resistant DT104 persisted at a mean percentage of 27%. DT104 was not recovered from the control chicks. Salmonella Ohio colonization levels tended to decline (r = 0.79, P > 0.05) over time in the control group (75 to 20%) and significantly decreased (P < 0.05) over time in both susceptible and resistant groups (40 to 10%, r = 0.82 and 55 to 5%, r = 0.85, respectively). Salmonella Senftenberg was recovered from the control group at low frequency throughout the trial and was not recovered from the other groups. For either trial, no apparent affect on morbidity or mortality was observed. Introduction of DT104 by commingling may induce colonization resulting in persistent high levels of shedding in flocks simultaneously with other Salmonella species. PMID- 11726147 TI - Distribution of Campylobacter spp. in selected U.S. poultry production and processing operations. AB - A study was conducted of 32 broiler flocks on eight different farms, belonging to four major U.S. producers. The farms were studied over I complete calendar year. Overall, 28 (87.5%) of the flocks became Campylobacter positive, and only four (12.5%) remained negative throughout the 6- to 8-week rearing period. In the majority of flocks, sampled every 2 weeks throughout production, Campylobacter positive fecal and cecal samples were not detected until 4 to 8 weeks of age. In only six of the flocks were environmental samples found to be positive before shedding of Campylobacter was detected in the birds. Even in some of the Campylobacter-negative flocks, contamination of the rearing environment was positive for Campylobacter but did not result in the birds subsequently excreting the organism. These findings are discussed in relation to U.S. husbandry practices and present uncertainty about sources of Campylobacter infection for poultry flocks. Birds were often transported to the processing plant in coops that were already contaminated with Campylobacter, and the organisms were sometimes found in samples of scald water and chill water. After chilling, the proportions of Campylobacter-positive carcasses from different producers ranged from 21.0 to 40.9%, which is lower than in other studies, and possible reasons are considered. PMID- 11726148 TI - Development of a selective differential agar for isolation and enumeration of Campylobacter spp. AB - Direct plating is an effective technique for isolation and enumeration of Campylobacters from a variety of sample types; however, distinguishing Campylobacters from non-Campylobacter contaminants that frequently grow on many existing agars is difficult. In this study, it was determined that exposing Campylobacters to low levels (200 mg/liter) of triphenyltetrazolium chloride (TTC) was not inhibitory to growth yet was sufficient to give a deep-red to magenta color to the colonies. The new agars (Campy-Line agar [CLAI and Campy Line blood agar [CLBA]) are translucent. The contrast of deep-red colonies on a translucent background greatly facilitates Campylobacter isolation and makes enumeration on light boxes or by electronic means possible. Direct plating of broiler carcass rinse samples (n = 20) was compared on Campy-Cefex agar and CLA. Recovery of Campylobacter populations was not significantly different between the agars (P < 0.05); however, enumeration was much less labor intensive with the CLA. No contaminants were observed on the CLA, whereas the Cefex agar supported the growth of approximately 14 contaminating (non-Campylobacter) CFU/ml. In a separate trial, recovery of Campylobacters from carcass rinses (n = 25) was similarly compared on Cefex, CLA, and CLBA. Again, recovery of Campylobacters was not significantly different between the agars (Pearson correlation coefficient = 0.988), whereas about nine contaminating (non-Campylobacter) CFU/ml were observed on Cefex agar and none on CLA or CLBA. Although some contaminants can still grow on CLA and CLBA and can present red colonies, most of these contaminants are easily distinguished from Campylobacter by differences in colony morphology. PMID- 11726149 TI - Levels of Vibrio vulnificus and organoleptic quality of raw shellstock oysters (Crassostrea virginica) maintained at different storage temperatures. AB - Temperature abuse during raw oyster harvesting and storage may allow for the multiplication of natural spoilage flora as well as microbial pathogens, thus posing a potential health threat to susceptible consumers and compromising product quality. The objective of this study was to provide a scientific basis for determining whether different refrigeration and abuse temperatures for raw oysters would result in a spoiled product before it became unsafe. Raw shellstock oysters (Crassostrea virginica) purchased from a commercial Virginia processor were subjected to different temperature abuse conditions (7, 13, and 21 degrees C) over a 10-day storage period. Salinity, pH, halophilic plate count (HPC), total culturable Vibrio counts, and culturable Vibrio vulnificus counts were determined at each abuse condition. V. vulnificus isolates were confirmed by a specific enzyme-linked immunosorbent assay. Olfactory analysis was performed to determine consumer acceptability of the oysters at each abuse stage. The pH of the oysters decreased over time in each storage condition. The HPC increased 2 to 4 logs for all storage conditions, while olfactory acceptance decreased over time. V. vulnificus levels increased over time, reaching 10(5) to 10(6) CFU/g by day 6. The length of storage had a greater effect on the bacterial counts and olfactory acceptance of the oysters (P < 0.05) over time than did the storage temperature (P < 0.05). PMID- 11726150 TI - Organic acids and their salts as dipping solutions to control listeria monocytogenes inoculated following processing of sliced pork bologna stored at 4 degrees C in vacuum packages. AB - Postprocessing contamination of cured meats with Listeria monocytogenes has become a major concern for the meat processing industry and an important food safety issue. This study evaluated aqueous dipping solutions of organic acids (2.5 or 5% lactic or acetic acid) or salts (2.5 or 5% sodium acetate or sodium diacetate, 5 or 10% sodium lactate, 5% potassium sorbate or potassium benzoate) to control L. monocytogenes on sliced, vacuum-packaged bologna stored at 4 degrees C for up to 120 days. Organic acids and salts were applied by immersing (1 min) in each solution inoculated (10(2) to 10(3) CFU/cm2) slices of bologna before vacuum packaging. Growth of L. monocytogenes (PALCAM agar) on inoculated bologna slices without treatment exceeded 7 log CFU/cm2 (P < 0.05) at 20 days of storage. No significant (P > 0.05) increase in L. monocytogenes populations occurred on bologna slices treated with 2.5 or 5% acetic acid, 5% sodium diacetate, or 5% potassium benzoate from day 0 to 120. Products treated with 5% potassium sorbate and 5% lactic acid were stored for 50 and 90 days, respectively, before a significant (P < 0.05) increase in L. monocytogenes occurred. All other treatments permitted growth of the pathogen at earlier days of storage, with sodium lactate (5 or 10%) permitting growth within 20 to 35 days. Extent of bacterial growth on trypticase soy agar plus 0.6% yeast extract (TSAYE) was similar to that on PALCAM, indicating that the major part of total bacteria grown on TSAYE agar plates incubated at 30 degrees C was L. monocytogenes. Further studies are needed to evaluate organic acids and salts as dipping solutions at abusive temperatures of retail storage, to optimize their concentrations in terms of product sensory quality, and to evaluate their effects against various other types of microorganisms and on product shelf life. In addition, technologies for the commercial application of postprocessing antimicrobial solutions in meat plants need to be developed. PMID- 11726151 TI - Reduction of listeria monocytogenes on green peppers (Capsicum annuum L.) by gaseous and aqueous chlorine dioxide and water washing and its growth at 7 degrees C. AB - Reduction of Listeria monocytogenes Scott A on uninjured and injured surfaces of green peppers after 0.3- and 3-mg/ liter gaseous and aqueous ClO2 treatment and water washing for 10 min at 20 degrees C was studied. Growth of the L. monocytogenes untreated or treated with 0.6 mg/liter ClO2 gas for 30 min at 20 degrees C on green peppers also was investigated. A membrane-surface-plating method was used for resuscitation and enumeration of L monocytogenes treated with ClO2. The bacterial viability on pepper surfaces was visualized using confocal laser scanning microscopy (CLSM). Live and dead cells of L. monocytogenes were labeled with a fluorescein isothiocyanate-labeled antibody and propidium iodide, respectively. More than 6 log CFU/5 g L. monocytogenes on uninjured surfaces and about 3.5 log CFU/5 g on injured surfaces were inactivated by both 3-mg/liter and 0.6-mg/liter ClO2 gas treatments. The 3-mg/liter aqueous ClO2 treatment achieved 3.7- and 0.4-log reductions on uninjured and injured surfaces, respectively; whereas, water washing alone showed 1.4- and 0.4-log reductions, respectively. ClO2 gas treatment was the most effective in reducing L. monocytogenes on both uninjured and injured green pepper surfaces, when compared with aqueous ClO2 treatment and water washing. The significant difference (P < 0.05) between log reductions on uninjured and injured surfaces and the results from CLSM analysis suggested that injured surfaces protected more bacteria from sanitation treatments than did uninjured surfaces. Not only could L. monocytogenes grow on green pepper surfaces at 7 degrees C, bacteria that survived the 0.6-mg/liter ClO2 gas treatment also could grow. PMID- 11726152 TI - Detection of heat injury in Listeria monocytogenes Scott A. AB - Methods of detecting live pathogens in foods that may be growth inhibited following heat treatment are essential to food safety. Among the techniques available, reverse transcription polymerase chain reaction (RT-PCR) amplification of messenger RNA from heat-injured Listeria monocytogenes Scott A is preferable to direct PCR in an attempt to avoid false positives from dead cells. The RT-PCR has a detection limit of 3 x 10(6) CFU/g, compared to 3 CFU/g for untreated controls, but may not be suitable for the identification of all viable cells. Physically apparent changes in cellular structures from heat injury in L. monocytogenes are expected to result. Ultrastructural analyses did depict notable heat damage as cytoplasmic clearing after 5 min at 60 degrees C. The heat-injured survivors can be readily distinguished from total viable cells using selective media. As a result, combinations of molecular and visual methods including selective media improve detectability of heat-injured, viable L. monocytogenes Scott A. PMID- 11726153 TI - Combination of immunomagnetic separation and polymerase chain reaction for the simultaneous detection of Listeria monocytogenes and Salmonella spp. in food samples. AB - A method that combined the immunomagnetic separation (IMS) technique and the multiplex polymerase chain reaction (PCR) method (i.e., the IMS-mPCR method) was developed for simultaneous detection of Listreria monocytogenes and Salmonella spp. in food samples. When only the multiplex PCR method was used, it was found that if cell numbers of each of the two target organisms (L. monocytogenes and Salmonella spp.) were above the detection limit, but differed by more than 2 logs e.g., n x 10(7) to n x 10(4) or n x 10(6) to n x 10(3)--the organism presenting the lower numbers might go undetected. Following the enrichment step with universal preenrichment (UP) broth, if an IMS method using equal quantities of anti-Listeria and anti-Salmonella immunomagnetic beads was performed prior to PCR, both pathogens could be detected unambiguously. Such results could be obtained for target organisms in food samples, such as milk, dairy, and meat products, if similar enrichment and IMS steps were performed prior to PCR. PMID- 11726154 TI - Evaluation of universal preenrichment broth for growth of heat-injured pathogens. AB - Universal preenrichment broth (UPB) was developed to enable enrichment of injured foodborne pathogens of different genera simultaneously in lieu of having to undergo separate simultaneous enrichment cultures for subsequent detection or isolation of each pathogen. Enrichment conditions in UPB for growth of injured pathogens to populations that will enable pathogen detection by rapid immuno based or polymerase chain reaction (PCR)-based assays have not been defined. Hence, studies were done to determine recovery and growth rates of heat-injured Escherichia coli O157:H7, Salmonella enterica ser. Typhimurium, Salmonella enterica ser. Enteritidis. and Listeria monocytogenes in UPB. Bacterial cells were heat injured in tryptic phosphate broth at 57.2 degrees C and inoculated at populations of ca. 0.17 to 63 injured cells per ml with raw ground beef, fresh chicken, lettuce, and environmental sponge samples. Enrichment cultures were sampled at 1, 2, 3, 4, 5, 6, and 24 h at 37 degrees C postinoculation, and pathogens were enumerated on appropriate selective media. Results revealed that recovery and growth of pathogens during the first 6 h of enrichment were not sufficient to ensure adequate numbers of bacteria (> 10(3) CFU/ ml) for detection by most immunoassays or PCR assays. Cells often required 3 to 4 h for recovery before growth was initiated. Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes cell populations in enrichment cultures with ground beef or lettuce at 6 h were 0.5 to 2.9 log10 CFU/ml. At 24 h of incubation, cell counts of enrichment samples for the three pathogens from all food and environmental sponge samples ranged from 4.0 to 8.3 log10 CFU/ml. Enrichment in UPB at 37 degrees C of foods or environmental sponge samples containing heat-injured cells of Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes reliably provides at 24 h of incubation-but not at 6 h-sufficient cell populations for detection by rapid immunoassay or PCR assay procedures that can detect at least 4 log10 CFU/ml. These results raise questions regarding the sensitivity of rapid detection methods that employ an abbreviated enrichment protocol of 6 h or less. PMID- 11726155 TI - Shellac formulations to reduce epiphytic survival of coliform bacteria on citrus fruit postharvest. AB - Survival of the coliform bacteria Enterobacter aerogenes and Escherichia coli was monitored in a neutral carboxymethylcellulose formulation and in shellac formulations with various pH and concentrations of ethanol and the preservative paraben; populations were subsequently measured from the surface of citrus fruit coated with these formulations. Numbers of the two bacteria increased over 24 h from 10(6) CFU/ml to approximately 10(8) CFU/ml in the carboxymethylcellulose solution, but over this time numbers remained little changed in the neutral solution of shellac. The Enterobacter was more tolerant of alcohol over a 3-h period: although its numbers in a shellac solution with 10% ethanol dropped from more than 10(6) CFU/ml to just over 10(3) CFU/ml. E. coli and a third species. Klebsiella pneunoniae, declined toward the limit of detection (5 CFU/ ml) during this time. The addition of morpholine to increase the formulation pH to 9.0 caused numbers of bacteria to plummet to an undetectable level within 30 to 60 min. On Ruby Red grapefruit and Valencia oranges in storage at 13 degrees C numbers of E. aerogenes and E. coli declined over 2 weeks from 10(5) CFU/cm2 to less than 2.5 x 10(1), but most of the loss in numbers occurred within 1 day. Numbers remained significantly less on shellacked fruit compared with those applied in the carboxymethylcellulose coating, and a shellac coating prepared from a pH 9 solution was more toxic to these species than one in which 12% ethanol had been added to the neutral formulation. The addition of the preservative paraben in the basic shellac was further inhibitory. PMID- 11726156 TI - Growth and recovery of selected gram-negative bacteria in reconditioned wastewater. AB - Previous reports indicate that Escherichia coli O157:H7, Salmonella spp., and Vibrio cholerae can grow in nutrient-limited, reconditioned wastewater over the temperature range of 4 to 46 degrees C when the biological oxygen demand of this water is <2, while its coliform growth response (CGR) is >2. In the current study, we investigated the growth response of Vibrio parahaemolyticus, Shigella spp., Vibrio vulnificus, and Pseudomonas aeruginosa in water samples with a CGR of >2 over the temperature range of 4 to 50 degrees C. Both the nonselective media, tryptic soy agar, and the selective media used to identify the pathogen were used for their recovery. The selective media were thiosulfate-citrate-bile sucrose (TCBS), MacConkey agar (MAC), and Pseudomonas isolation agar (PIA) for the Vibrio, Shigella, and Pseudomonas spp., respectively. V. parahaemolyticus numbers declined rapidly after surviving for 6 days under the nutrient-limiting growth conditions. Shigella spp. did not grow but survived for >28 days at 4 to 25 degrees C. V. vulnificus grew over the narrow temperature range of 12 to 21 degrees C and survived for >21 days at the higher and lower temperature ranges. P. aeruginosa survived and grew during the 14-day test period at 13 to 35 degrees C. Recovery on the nonselective agar gave statistically (P > 0.05) higher numbers than the respective selective media commonly used for these pathogens. These results indicate that caution should be used in attempting direct recoveries using selective media of the four gram-negative bacteria species used in this study from the nutrient-limited water environment. PMID- 11726157 TI - Effect of Lactobacillus casei and yogurt administration on prevention of Pseudomonas aeruginosa infection in young mice. AB - Pseudomonas aeruginosa is an opportunistic pathogen that rarely causes pulmonary disease in normal hosts but one that is an important cause of acute pneumonia in immunocompromised patients, including neonates, and of chronic pneumonia in patients with cystic fibrosis. The aim of this work was to study the effect of oral administration of Lactobacillus casei and yogurt on prevention of P. aeruginosa lung infection in young mice (3 weeks old). This study demonstrates that oral administration of L. casei or yogurt to young mice enhanced lung clearance of P. aeruginosa and phagocytic activity of alveolar macrophages through a dose-dependent effect. There were, however, no significant differences in white blood cell (WBC) differential counts. Furthermore, it was observed that previous administration of L. casei or yogurt induced a significant increase in IgA and IgM levels in bronchoalveolar lavages (BALs) after a P. aeruginosa infection, although there was no relationship with the serum values. PMID- 11726158 TI - Growth and viability of commercial Bifidobacterium spp. in honey-sweetened skim milk. AB - Two commercial Bifidobacterium bifidum (Bf-1 and Bf-6) were cultured in 12% (wt/vol) reconstituted nonfat dry milk (NDM) containing 5% (wt/wt) honey, sucrose, fructose, or glucose. Inoculated samples were incubated anaerobically at 37 degrees C for 48 h. Samples were collected at 12-h intervals and examined for (i) specific growth rate, (ii) pH, and (iii) levels of fermentation end products (lactic and acetic acids) as measured by high-performance liquid chromatography (HPLC). Viability of the organisms during 28 days of refrigerated storage at 4 degrees C was also assessed at 7-day intervals. Growth promotion and acid production were greatest when Bf-1 and Bf-6 were grown in the presence of honey. For both Bf-1 and Bf-6, retention of viability was greatest up to 14 days of refrigerated storageat 4 degrees C when they were grown and stored in the presence of honey compared to other sweeteners. PMID- 11726159 TI - Toxicity of moniliformin from Fusarium fujikuroi culture material to growing barrows. AB - In two studies, the effects of moniliformin (M)-contaminated diets from Fusarium fujikuroi culture material on growing barrows were evaluated. In the first study, six barrows (three replicates of two each, mean body weight = 17.8 kg) per group (four groups; 24 barrows total) were fed diets calculated to contain 0 mg M/kg feed (control); 25 mg M/kg feed; 50 mg M/kg feed; or 100 mg M/kg feed for 28 days. In the second study, the same experimental design and numbers of barrows (mean body weight = 15.3 kg) were used, and diets were formulated to contain 0 mg M/kg feed (control); 50 mg M/kg feed; 100 mg M/kg feed; or 200 mg M/kg feed. Diets of 100 mg or 200 mg M/kg feed reduced body weight, body weight gain, and feed consumption. Serum biochemical analytes were affected by 100 to 200 mg M/kg feed. Hematologic values were affected by 50, 100, and 200 mg M/kg feed. In the first study, one barrow in the 100 mg M-treated group died, and in the second study. one barrow died in the 100 mg M-treated group, and five barrows died in the 200 mg M-treated group. Relative heart weight was increased in the 200 mg M treated barrows, yet tissues from organs collected from treatment groups were generally histologically unimpressive. The most consistent sign of M toxicity in barrows appeared to be death induced within 2 to 5 days by 100 to 200 mg M/kg feed. PMID- 11726160 TI - Comparison of constitutive and inducible maize kernel proteins of genotypes resistant or susceptible to aflatoxin production. AB - Maize genotypes resistant or susceptible to aflatoxin production or contamination were compared for differences in both constitutive and inducible proteins. Five additional constitutive proteins were found to be associated with resistance in over 8 of the 10 genotypes examined. Among these, the 58- and 46-kDa proteins were identified as globulin-1 and globulin-2, respectively. Differences in the ability to induce specific antifungal proteins, such as the higher synthesis of the 22-kDa zeamatin in resistant genotypes, were also observed between resistant and susceptible kernels incubated under germinating conditions (31 degrees C, 100% humidity). Both constitutive and inducible proteins appear to be necessary for kernel resistance. Embryo-killed kernels (unable to synthesize new proteins) supported the highest level of aflatoxins, whereas imbibed kernels (to hasten protein induction) supported the lowest among all treatments. This suggests that the synthesis of new proteins by the embryo plays an important role in conferring resistance. However, significantly lower levels of aflatoxin production in embryo killed resistant kernels than in susceptible ones suggest that, in reality, high levels of constitutive antifungal proteins are indispensable to kernel resistance. PMID- 11726161 TI - Occurrence of parasites on fruits and vegetables in Norway. AB - Between August 1999 and January 2001, samples of various fruits and vegetables obtained within Norway were analyzed by published methods for parasite contamination. Neither Cyclospora oocysts nor Ascaris (or other helminth) eggs were detected on any of the samples examined for these parasites. However, of the 475 samples examined for Cryptosporidium oocysts and Giardia cysts, 29 (6%) were found to be positive. No samples were positive for both parasites. Of the 19 Cryptosporidium-positive samples. 5 (26%) were in lettuce, and 14 (74%) in mung bean sprouts. Of the 10 Giardia-positive samples, 2 (20%) were in dill, 2 (20%) in lettuce, 3 (30%) in mung bean sprouts, 1 (10%) in radish sprouts, and 2 (20%) in strawberries. Mung bean sprouts were significantly more likely to be contaminated with Cryptosporidium oocysts or Giardia cysts than the other fruits and vegetables. Concentrations of Cryptosporidium and Giardia detected were generally low (mean of approximately 3 [oo]cysts per 100 g produce). Although some of the contaminated produce was imported (the majority, if sprouted seeds are excluded), there was no association between imported produce and detection of parasites. Crvptosporidium oocysts and Giardia cysts were also detected in water samples concerned with field irrigation and production of bean sprouts within Norway. This is the first time that parasites have been detected on vegetables and fruit obtained in a highly developed. wealthy country, without there being an outbreak situation. These findings may have important implications for global food safety. PMID- 11726162 TI - Factors affecting recovery efficiency in isolation of Cryptosporidium oocysts and Giardia cysts from vegetables for standard method development. AB - While recently published techniques for recovering parasites from fruits and vegetables demonstrate a marked increase in efficiency and utility, there is still scope for further improvement in developing a standard method, particularly with difficult, but important, sample matrices such as bean sprouts. Herein, a number of parameters used in published techniques are investigated more closely. While sample size reduction may improve recovery efficiency because of a range of factors, it is important to keep the sample large enough for detection of low level contamination. Age of sample is also important, and samples should be as fresh as possible. Elution procedures may contribute to losses of Giardia and should be more thoroughly investigated. Improved immunomagnetic separation techniques currently coming onto the market also have the potential to increase recovery efficiency substantially, even with difficult samples such as aged bean sprouts. However, merely increasing magnetic strength of the capturing magnet does not affect recovery efficiency, which must be reliant on a superior bead system, buffering system, or both. PMID- 11726163 TI - Sucrose, sodium dodecyl sulfate, urea, and 2-mercaptoethanol affect the thermal inactivation of R-phycoerythrin. AB - Thermal inactivation kinetics (D- and z-values) of the algal protein, R phycoerythrin (R-PE), were studied under different buffer conditions (pH 4.0, 7.0, and 10.0) and concentrations of sucrose, sodium dodecyl sulfate (SDS), urea, and 2-mercaptoethanol (ME). R-PE solutions were heated in capillary tubes at temperatures between 40 and 90 degrees C depending on buffer conditions. Thermal inactivation parameters for R-PE, calculated on the basis of fluorescence loss, were modified by addition of chemicals. Overall, sucrose and ME had a thermostabilizing effect, while SDS and urea decreased thermal stability of R-PE. The z-values ranged from 5.9 degrees C in 50 mM NaCl, 20 mM glycine buffer, pH 10.0, to 37.8 degrees C in 60% sucrose, 50 mM NaCl, 20 mM phosphate buffer, pH 7.0. The z-values obtained for R-PE closely matched the z-values of some target microorganisms in food processes, suggesting R-PE might be used as a time temperature integrator to verify thermal processing adequacy. PMID- 11726164 TI - Development of a sandwich enzyme-linked immunosorbent assay for the detection of egg residues in processed foodst. AB - Chicken eggs are used extensively as an excellent source of dietary proteins. These proteins have many functional properties, making them valuable food ingredients. However, eggs are a frequent cause of food hypersensitivity, especially in children. Of major concern to food processors is the inadvertent cross-contact of food products with allergenic residues, which could result in potentially life-threatening reactions in those with a food allergy. The aim of the present study was to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of undeclared egg residues in foods. Commercially purified ovalbumin (OVA) and dehydrated egg white solids were used as antigens to induce antibodies in rabbits and goats. Reference pasta standards and various food samples were extracted, then clarified by centrifugation. Goat anti-egg white antibodies were used as the capture reagent, nonspecific sites were blocked with gelatin, then standard and sample extracts were added. Rabbit anti-OVA antibodies were used as detector antibodies, followed by addition of commercial goat anti rabbit IgG antibody labeled with alkaline phosphatase and subsequent substrate addition. Twenty brands of egg-free pasta (two lots each) were analyzed using the ELISA. Fourteen common pasta ingredients were also evaluated for cross-reactivity problems in the method. The detection limit of the assay was 1 ppm spray-dried whole egg. Fifty-five percent (22 samples) of the egg-free pasta samples tested positive for the presence of undeclared egg residues, with values ranging from 1 to >100,000 ppm. Minimal cross-reactivity was encountered in general, but portobello mushrooms and basil caused some minor matrix effects. This sandwich type ELISA method can be used to detect undeclared egg residues in processed foods and to evaluate industrial clean-up operations. PMID- 11726165 TI - Reduction of Escherichia coli O157:H7 and Salmonella typhimurium in artificially contaminated alfalfa seeds and mung beans by fumigation with ammonia. AB - Sprouts eaten raw are increasingly perceived as hazardous foods because they have been vehicles in outbreaks of foodborne disease, often involving Escherichia coli O157:H7 and Salmonella Typhimurium. Although the source of these pathogens has not been established, it is known that the seeds usually are already contaminated at the time sprouting begins. Earlier studies had shown that ammonia was lethal to these same pathogens in manure, so it seemed reasonable to determine whether ammonia was effective against them when associated with seeds to be used for sprouting. Experimentally contaminated (10(8) to 10(9) CFU/g) and dried seeds, intended for sprouting, were sealed in glass jars in which 180 or 300 mg of ammonia/liter of air space was generated by action of ammonium sulfate and sodium hydroxide. Samples were taken after intervals up to 22 h at 20 degrees C. Destruction of approximately 2 to 3 logs was observed with both bacteria associated with alfalfa seeds, versus 5 to 6 logs with mung beans. Greater kills are apparently associated with lower initial bacterial loads. Germination of these seeds was unaffected by the treatment. It appears that this simple treatment could contribute significantly to the safety of sprout production from alfalfa seeds and mung beans. PMID- 11726166 TI - Penetration of Escherichia coli O157:H7 into lettuce as influenced by modified atmosphere and temperature. AB - The effects of temperature and atmospheric oxygen concentration on the respiration rate of iceberg lettuce and Escherichia coli O157:H7 cells attachment to and penetration into damaged lettuce tissues were evaluated. Respiration rate of lettuce decreased as the temperature was reduced from 37 to 10 degrees C. Reducing the temperature further to 4 degrees C did not affect the respiration rate of lettuce. Respiration rate was also reduced by lowering the atmospheric oxygen concentration. Lettuce was submerged in E. coli O157:H7 inoculum at 4, 10, 22, or 37 degrees C under 21 or 2.7% oxygen. Attachment and penetration of E. coli O157:H7 were not related to the respiration rate. The greatest numbers of E. coli O157:H7 cells attached to damaged lettuce tissues at 22 degrees C at both oxygen concentrations. More cells were attached under 21% oxygen than under 2.7% oxygen at each temperature, but this difference was small. Penetration of E. coli O157:H7 into lettuce tissue was determined by immunostaining with a fluorescein isothiocyanate-labeled antibody. Under 21% oxygen, E. coli O157:H7 cells showed greatest penetration when lettuce was held at 4 degrees C, compared to 10, 22. or 37 degrees C, and were detected at an average of 101 microm below the surfaces of cut tissues. However, under 2.7% oxygen, there were no differences in degree of penetration among four incubation temperatures. The degree of E. coli O157:H7 penetration into lettuce tissue at 4 or 22 degrees C was greater under 21% oxygen than under 2.7% oxygen; however, no difference was observed at 37 degrees C. Conditions that promote pathogen penetration into tissue could decrease the effectiveness of decontamination treatments. PMID- 11726167 TI - Contamination of beef chucks with Escherichia coli during carcass breaking. AB - Samples were obtained by swabbing the whole of the chuck portion on each of the first 500 sides that entered a beef carcass breaking process and the whole of the outer surface of each of the chuck primal cuts that were prepared from those portions. Swabs obtained from groups of 10 sides or cuts that entered or emerged from the process consecutively were combined, and the coliforms and Escherichia coli recovered from each group were enumerated. Coliforms and E. coli were recovered only sporadically from groups of sides at log total numbers of 4.0 and 3.5 log CFU/500 sides, respectively. Coliforms were recovered from three and E. coli from none of the first six groups of cuts. Coliforms and E. coli were recovered from all subsequent groups of cuts, initially at log numbers mostly <3 log CFU/10 cuts, but ultimately at log numbers mostly >3 log CFU/10 cuts. The log total numbers of coliforms and E. coli recovered from cuts were >6.0 and 5.5 log CFU/500 cuts, respectively. After the breaking of about 600 sides, samples were obtained by swabbing a table onto which the part of the side that included the chuck portion was deposited after it was cut from the hanging side, and the belt that was used for conveying chucks. The numbers of coliforms and E. coli recovered from the table and conveyor belt were comparable with the numbers recovered from sides and cuts, respectively. Those findings show that most of the coliforms and E. coli recovered from the cuts were not present on carcass sides but that they originated largely from the cut conveying equipment. PMID- 11726168 TI - Growth rates of Salmonella and Escherichia coli O157:H7 in irradiated beef. AB - Ground beef was irradiated to 0, 2, or 4 kGy and then inoculated with a mixed culture of four serotypes of salmonellae or five strains of Escherichia coli O157:H7. The ground beef was stored at either 15 or 25 degrees C, and the growth of the inoculated bacteria was monitored over time. Growth parameters were determined for both the salmonellae and the E. coli O157:H7 using the Gompertz equation. There was no significant difference in lag phase duration or generation time, irrespective of the dose to which the ground beef had previously been exposed. Furthermore, the lag phase durations and generation times determined in this study did not differ significantly from previously published values. This suggests that, although irradiation eliminates a significant portion of the spoilage microflora in ground beef, the absence of this microflora provides no competitive advantage to the growth of salmonellae or E. coli O157:H7 in ground beef. PMID- 11726169 TI - Prevalence of Salmonella serovars in chickens in Turkey. AB - In this study, 151 (18.6%) of 814 ceca obtained during in-line processing of 28 broiler (Hybro G, Avian, Arbor acres, and Cobb breeds) and 5 layer (Ross, Tetra SL, Isa Brown, and Brown Nick breeds) flocks in Turkey were found to be contaminated with four different Salmonella serovars. Only Salmonella enterica subsp. enterica Serovar Enteritidis (Salmonella Enteritidis) was recovered from layer birds, whereas Salmonella Enteritidis (81.5%). Salmonella Agona (7.6%), Salmonella Thompson (10.1%), and Salmonella Sarajane (0.8%) were isolated from broiler birds. Isolations of Salmonella Agona and Salmonella Thompson from poultry are reported for the first time in Turkey. The isolation of Salmonella Sarajane from chickens is the first report in the world. The standard method of National Poultry Improvement Plan, U.S. Department of Agriculture, was used to detect Salmonella from chicken cecal samples. Primary and delayed secondary enrichments (PE and DSE) were done in tetrathionate-Hajna broth (TTHB). Two different agar media, xylose lysine tergitol 4 (XLT4) and brilliant green with novobiocin (BGN) were used to observe, and compared for their isolation and selective differentiation of, Salmonella-suspected colonies. Isolated salmonellae were then biotyped and serotyped. Ninety-one and 151 salmonellae were isolated with XLT4 agar after PE and DSE, respectively. From the same samples, BGN agar was able to detect only 50 and 131 Salmonella after PE and DSE, respectively. The isolation rate with XLT4 was 11.2% (P < 0.01) with PE, and this rate increased to 18.6% after DSE. Also, the PE isolation rate (11.2%) with XLT4 agar was significantly higher (P < 0.01) than PE with BGN agar (6.1%). Salmonella was isolated from 39.3% (11 of 28) of the broiler flocks and from 60.0% (3 of 5) of the layers. The detection sensitivity of the isolation method was determined as 1 CFU g(-1) experimentally. These data demonstrate the presence of Salmonella Enteritidis, Salmonella Thompson, Salmonella Agona, and Salmonella Sarajane in chicken flocks in Turkey. PMID- 11726170 TI - Behavior of aeromonas hydrophila in bottled mineral waters. AB - The growth and survival of Aeromonas hydrophila in three types of natural mineral waters were investigated. Mineral waters with different levels of mineral content (low, medium, and high) were experimentally contaminated with A. hydrophila, stored at different temperatures (10 degrees C and 20 degrees C), and analyzed at intervals over a 60-day period. Water samples that were not experimentally contaminated were investigated for indigenous A. hydrophila. The results confirmed that A. hydrophila may occur naturally in mineral waters and showed that the level of mineral content, temperature, length of storage, and, in some cases, the type of container used may favor the growth of A. hydrophila. The greatest proliferation was observed in water with a low mineral content stored in PET bottles at 10 degrees C, in which A. hydrophila peaked at day 28 (4.47 +/- 0.01 log CFU/100 ml). At 20 degrees C, the same load was observed at day 60. The presence of high densities of A. hydrophila in bottled mineral water can constitute a risk for some groups of consumers, such as elderly and immunocompromised persons. PMID- 11726171 TI - Evaluation of the petrifilm plate method for the enumeration of aerobic microorganisms and coliforms in retailed meat samples. AB - This study was designed to compare the effectiveness and applicability of the Petrifilm plate method with the Association of Official Analytical Chemists' (AOAC) standard aerobic count method and violet red bile agar method for meat products. The comparison was carried out using 303 meat samples collected from various retailers: 110 pork samples, 87 chicken samples, and 107 beef samples. In the comparison of the correlation coefficient (R) between the conventional method and the Petrifilm plate method by a linear regression analysis, the correlation coefficient in total microorganisms was 0.99, 0.95, and 0.94 in pork, beef, and chicken samples, respectively. The correlation coefficient in coliform count was 0.83, 0.96, and 0.81 in pork, beef, and chicken samples, respectively. Based on the high correlation in the total microorganism count, it might be possible to replace the conventional methods with the Petrifilm plate method. For coliform counts, the Petrifilm plate method also showed a generally high correlation coefficient, except for pork samples, which are more subject to contamination. The Petrifilm plate method was simpler and less time-consuming in sample preparation and, in procedures, faster than the conventional method. These results suggested that the 3M Petrifilm plate method could replace the conventional methods in the analysis of microorganism contamination measurement in meat products. PMID- 11726172 TI - Detection limits of beta-lactam antibiotics in ewe milk by penzym enzymatic test. AB - The Penzym is an enzymatic test widely used for the detection of beta-lactam antibiotic residuals in milk. It is a specific method with good sensitivity to this group of antibiotics and enables results to be obtained within a short time. In the present work, the detection limits of 10 beta-lactam antibiotics were determined in ewe milk, given the lack of previous studies of the Penzym test in ovine milk. For each antibiotic, eight concentrations were tested on 20 ewe milk samples proceeding from individual ewes (160 analyses per drug). The limits of the Penzym test were determined by means of logistic regression models, as follows: 5 microg/kg amoxicillin, 4 microg/kg ampicillin, 33 microg/kg cloxacillin, 3 microg/kg penicillin "G," 43 microg/kg cephadroxil. 10 microg/kg cephalosporin "C," 16 microg/kg cephalexin, 900 microg/kg cephoperazone, 120 microg/kg Ceftiofur, and 77 microg/kg cephuroxime. The percentages of positive results for those antibiotics at the maximum residue limit (MRL) concentration established by the European Union (EU) were: 100% (penicillin "G"), 93.3% (ampicillin), 93.3% (cloxacillin), 56.7% (Ceftiofur), and 56.7% (amoxicillin). PMID- 11726173 TI - Determination of deoxynivalenol in wheat-based breakfast cereals marketed in Portugal. AB - Deoxynivalenol (DON), also known as vomitoxin, is one of a group of closely related secondary fungal metabolites--the trichothecenes--and is produced predominantly by several species of the genus Fusarium, especially Fusarium graminearum. The present study was carried out to evaluate the natural occurrence of DON in different kinds of wheat-based breakfast cereals widely consumed by the population. A total of 88 commercially available samples of wheat-based breakfast cereals were randomly collected from different supermarkets in Lisbon, Portugal. The samples were analyzed using immunoaffinity column, and DON was quantified by liquid chromatography. Detection limit was 100 microg/kg. Average recovery of DON was 80%. Of 88 analyzed samples, 72.8% contained levels of DON between 103 and 6,040 microg/kg, with mean level of 754 microg/kg, and 24 samples (27.2%) were not contaminated (< 100 microg/kg). These results indicate an incidence of this mycotoxin in these products, and the authors suggest a monitoring for the prevention of molds and mycotoxins. This is the first report in Portugal on natural contamination with DON in wheat-based breakfast cereals. PMID- 11726174 TI - Incidence of patulin in apple juices marketed in Turkey. AB - The purpose of this study was to investigate the patulin contamination of apple juices consumed by the Turkish population. Patulin was detected using high performance liquid chromatography (HPLC) with a UV detector at 280 nm, and the identification of patulin was further confirmed by thin-layer chromatography (TLC). Using HPLC, the recoveries were 79.9 +/- 6.7% and 83.7 +/- 4.6%, and the coefficients of variation were 8.4 and 5.5% for apple juices spiked with the known amounts of patulin (60 and 120 microg/liter. respectively). The minimum patulin level detected was 5 ng in a standard solution and 5 microg/liter in apple juices. The TLC method was used only to confirm patulin levels higher than 20 microg/liter (100 ng/spot) in apple juices. The total number of samples was 45. Patulin was present in detectable levels in 60% of apple juices at concentrations ranging from 19.1 to 732.8 microg/liter. Forty-four percent of the apple juice samples had patulin contamination levels higher than 50 microg/ liter, which is the allowable upper limit in Turkey. PMID- 11726176 TI - Total, organic, and inorganic arsenic in some commercial species of crustaceans from the Mediterranean Sea (Italy). AB - Total, organic, and inorganic arsenic were measured in the flesh of different crustacean species: Parapenaeus longirostris, Aristeus antennatus, Plesionika martia, and Nephrops norvegicus, species extensively commercialized and widely consumed as food. Among the different species of crustaceans, the highest mean value of total arsenic was detected in N. norvegicus (45.00 microg g(-1) dry wt), followed by P. martia (40.76 microg g(-1) dry wt), P. longirostris (34.00 microg g(-1) dry wt), and A. antennatus (15.45 microg g(-1) dry wt). In all the species examined, most of this element was present as organic compounds, while inorganic arsenic was only a very modest percentage (3.5 to 5.7%) of the total arsenic present. There is a substantial epidemiological evidence that inorganic arsenic may cause lung and skin cancer. For this reason, the Joint Expert Committee of the FAO-WHO recommended a maximum acceptable daily intake of inorganic arsenic for humans of 2 microg/kg of body weight. The estimated daily intake was below this limit; therefore, no direct hazard for human health due to the consumption of this seafood was found. PMID- 11726175 TI - Coccidial contamination of raspberries: mock contamination with Eimeria acervulina as a model for decontamination treatment studies. AB - Numerous outbreaks have been reported since 1995 in the United States and Canada that were linked to the consumption of imported fresh raspberries contaminated with Cyclospora. Because Cyclospora has no laboratory animal hosts, Eimeria acervulina, a common chicken coccidium similar in characteristics to Cyclospora, was used as a surrogate to test decontamination treatments. Raspberries were mock contaminated with E. acervulina-sporulated oocysts in a water suspension, then exposed to washing, freezing, heat, or irradiation before they were fed to chicks. The presence of oocysts in the contaminated raspberries was confirmed either by duodenal lesions or oocysts in cecal contents 5 days postinoculation (PI) or in fecal contents 6 days PI, after 24 h of fecal collection. Washing of raspberries was generally not adequate in removing coccidial contamination, but freezing and heat treatment appeared effective. Gamma irradiation of E. acervulina-sporulated oocysts at a dose of 0.5 kGy was partially effective, but it was completely effective at 1.0 kGy and higher. We suggest that E. acervulina, for mock contamination of raspberries and subsequent decontamination treatments, is easy to handle, safe, and economical to study. PMID- 11726177 TI - Foodborne outbreaks in Canada linked to produce. AB - Examples of foodborne outbreaks traced to fresh fruits and vegetables can be found worldwide. The quantity of produce eaten per capita has been increasing steadily over the past two decades, creating a heightened potential for produce related foodborne disease. A number of outbreaks identified during this time period were reviewed, with particular emphasis placed on incidents that have occurred in Canada. The collective information highlights the diversity of infectious agents and produce items involved, with a view to the prevention of fresh produce-related foodborne disease in the future. PMID- 11726178 TI - Kinetic locking-on strategy and auxiliary tactics for bioaffinity purification of NAD(P)(+)-dependent dehydrogenases. PMID- 11726179 TI - An isothermal titration calorimetric method to determine the kinetic parameters of enzyme catalytic reaction by employing the product inhibition as probe. AB - An isothermal titration calorimetric (ITC) method was developed to measure the kinetic parameters of ribonuclease A catalytic hydrolysis of cytidine 2',3' cyclic monophosphate. Employing the inhibition of product as a probe, the K(m), K(i), k(c), and DeltaH(m) can be determined by two simple calorimetric measurements. First, the substrate was titrated into the cell containing high concentration of enzyme. The molar reaction heat was calculated from the titration peak area divided by substrate moles per titration, and the initial catalytic reaction rate in the presence of various concentrations of product can be calculated from the peak height and the molar reaction heat. From Michaelis Menten function in the presence of inhibitors, the relationship between K(m) and K(i) can be obtained. Then, the dissociation constant, which is equal to K(i), was measured by a regular ITC experiment. Thus, K(m) and k(c) can be calculated. The method developed here can be applied in other enzyme catalytic systems with inhibitive products. PMID- 11726180 TI - Detection of single base alterations in genomic DNA by solid phase polymerase chain reaction on oligonucleotide microarrays. AB - DNA microarray technology holds significant promise for human DNA diagnostics. A number of technical approaches directed at the parallel identification of mutations or single nucleotide polymorphisms make use of polymerase-based specificity, like minisequencing or allele-specific primer elongation. These techniques, however, require separate laborious sample amplification, preparation, and purification steps, making large-scale analyses time and cost consuming. Here, we address this challenge by applying an experimental setup using simultaneous solid and liquid phase PCR on polyethyleneimine-coated glass slides, a novel microarray support allowing on-chip amplification reactions with exquisite specificity. A gene-specific oligonucleotide tiling array contains covalently attached allele-specific primers which interrogate single nucleotide positions within a genomic region of interest. During a thermal cycling reaction amplification products remain covalently bound to the solid support and can be visualized and analyzed by the incorporation of fluorescent dyes. Using the described procedure we unequivocally defined the presence of point mutations in the human tumor suppressor gene p53 directly from a natural DNA source. This semi multiplex solid phase amplification format allowed the rapid and correct identification of 20 nucleotide positions from minute amounts of human genomic DNA. Our results suggest that this approach might constitute a vital component of future integrated DNA chip devices used in gene analysis. PMID- 11726181 TI - A method for identification of inhibitors of the phosphorylation reactions of bacterial response regulator proteins using (31)P nuclear magnetic resonance spectroscopy. AB - Bacterial response regulators are attractive targets for antibacterial drug development, yet random screening against these targets has failed as yet to identify chemicals that constitute viable leads. Alternative methods to provide leads for drug development based on identification and optimization of low affinity ligands from NMR screens have been described. However, leads from these processes still require verification in a bioassay, which is often problematic if compounds have unfavorable optical and solubility properties. A simple method, based on using NMR to observe the activity of the target, is described. It has the advantages of being able to characterize both low affinity leads and a wider selection of compounds in a structure activity relationships series, without the problems affecting a fluorescence assay. In this example we use (31)P to monitor the turnover of a bacterial response regulator, but the generic approach could be applied to other nuclei and thus a range of biological systems. PMID- 11726182 TI - An automated high-performance liquid chromatography fluorescence method for the analyses of endothelins in plasma samples. AB - A high-performance liquid chromatographic method with fluorescence detection was developed to simultaneously analyze endothelins, a class of vasoactive peptides, in plasma samples. Sample preparation for HPLC analysis was carried out by initial stabilization of blood and plasma samples against transformation of big endothelins to mature endothelins and breakdown of mature endothelins by serine proteases, as well as oxidative modifications of endothelins. Deproteinization of plasma samples was achieved with acidified acetone, and the samples were further purified on molecular weight cutoff filters. Endothelins were separated on a reversed phase LC-318 column by gradient elution using a mobile phase containing acetonitrile and water (0.1% trifluoroacetic acid) and were analyzed by fluorescence detection (lambda(Ex), 280 nm; lambda(Em), 340). Limit of detection values were in the range of 0.2-0.5 pmol. Linear (R(2), 0.99) calibration curves were established for analyte amounts in the range of 1 to 100 pmols. Recoveries of endothelins from spiked plasma samples analyzed ranged from 60-95%. Under optimized conditions the HPLC-fluorescence method was determined to be sensitive and specific for the analysis of big endothelin-1, endothelin-1, endothelin-2, and endothelin-3 in plasma. Simultaneous measurement of these endothelins by the HPLC method should permit a better understanding of their specific roles and relationships under various pathological conditions. PMID- 11726183 TI - Development and validation of a competitive AKT serine/threonine kinase fluorescence polarization assay using a product-specific anti-phospho-serine antibody. AB - A competitive fluorescence polarization (FP) assay has been developed for the serine/threonine kinase, AKT. The FP assay has been formatted in a 384-well microtiter plate and automated using a pipeting workstation with performance suitable for high-throughput screening. The assay design utilizes a fluorescent phosphorylated peptide complexed to a product-specific anti-phospho-serine antibody. When unlabeled substrate is phosphorylated, by the kinase, the product competes with the fluorescent phosphorylated peptide for the antibody. The fluorescent phosphorylated peptide is then released from the antibody into solution resulting in a loss in polarization signal. Seven fluorescent phosphorylated peptides and 19 antibodies were evaluated for this assay. RARTSpSFAEPGK-Fl peptide and anti-phospho-GSK-3alpha Ser21 antibody gave the best affinity and change in polarization signal. The apparent kinetic constants were calculated for the FP assay and were consistent with reported values. The FP assay was validated with known inhibitors and the results compared to a radioactive Flashplate transfer assay, utilizing [(33)P]ATP and a biotinylated substrate, also developed in our laboratory. The IC(50) values generated were comparable between the two methods suggesting the competitive FP assay and Flashplate assay have similar sensitivities and abilities to identify inhibitors during screening. PMID- 11726184 TI - Development of sensitive esterase assays based on alpha-cyano-containing esters. AB - A novel approach is reported for the development of fluorogenic esterase reporters using alpha-cyano-containing esters as substrates. After ester hydrolysis, the released alcohol, a cyanohydrin, rapidly eliminates HCN to yield the corresponding aldehyde resulting in strong fluorescence. The pi conjugation of the resulting aldehyde also greatly enhances UV absorption and red shifts fluorescence emission relative to a corresponding alcohol or phenol. Two substrates, R/S-acetic acid cyano-(6-methoxynaphthalen-2-yl)-methyl ester (compound I) and trans/cis-3-(2,2-dichlorovinyl)-2,2 dimethylcyclopropanecarboxylic acid R/S-cyano-(6-methoxynaphthalen-2-yl)-methyl ester (compound II), were synthesized and evaluated as substrates. Such alpha cyano substrates possess very low background fluorescence and are more stable under enzyme assay conditions than phenolic substrates due to the aliphatic cyano group. The higher molar absorbtivity and quantum yield of the aldehyde, along with its larger Stokes' shift combined with the increased stability and lower background signal of the cyanohydrin substrate, increases the utility and sensitivity of the resulting assays over current methods. Moreover, compound II showed high selectivity to pyrethroid-cleaving esterases and may provide a direct tool to monitor pyrethroid resistance in insects. PMID- 11726185 TI - Identification and validation of endogenous reference genes for expression profiling of T helper cell differentiation by quantitative real-time RT-PCR. AB - Real-time RT-PCR method was exploited to identify endogenous reference genes in differentiating human T helper cells. When using this technology in our experimental system, finding a set of genes whose mRNA expression levels would not change appeared to be very challenging. Our initial plan to use the expression level of GAPDH in normalizing the results failed, because the mRNA expression of GAPDH underwent significant changes during the cell culture. Additional studies on the transcription of several other classical housekeeping genes led to similar results. Our second approach was to use results from an extensive survey of gene expression done by oligonucleotide microarrays and to select another panel of genes for testing. This resulted in the identification of three genes whose expression was relatively stable in our experimental system and, therefore, suitable as endogenous reference genes in these cells. The results indicate that the expression level of a constitutively expressed gene may change during the cell culture in vitro, which emphasizes again the importance of carefully validating endogenous control genes for comparative quantification. PMID- 11726186 TI - Studies of a disposable biosensor based on the beta-cyclodextrin inclusion complex as mediator. AB - This paper reports the use of a disposable voltammetric biosensor based on a screen-printed basal electrode and highly efficient mediators, the beta cyclodextrin inclusion complexes tetramethylbenzidine and ferrocene. The polyphenol oxidase, contained in the crude extract of mushroom, was immobilized in the membrane of the cross-linked beta-cyclodextrin polymer. The resulting biosensor gave excellent analytical performance due to the supramolecular complexation for mediators. It responded to the concentration of dopamine with high selectively in the range from 1.0 x 10(-9) to 1.0 x 10(-6) M with a detection limit as low as 5 x 10(-10) M. The response time for reaching 90% of its steady-state value was less than 60 s, and the activity was retained for at least 15 days. PMID- 11726187 TI - Synthesis and purification of 3-hydroxykynurenine-O-beta-glucoside, a primate lens ultraviolet filter, and its application in a two-step assay for beta glucosidase activity. AB - 3-Hydroxykynurenine-3-O-beta-glucoside (3-HKG) functions in the primate lens as a filter of 295- 400-nm light, thereby protecting the retina from damaging UV radiation. Although extensive studies have been conducted to determine the functional role of 3-HKG in the primate lens, an efficient method for its synthesis and purification has yet to be developed. Several procedures have been reported for the synthesis of 3-HKG; however, these procedures either result in low yields or require numerous sequential reactions and purification steps. In this study, we report a two-step synthesis of 3-HKG with a one-step purification and a two- to eightfold increase in yield over previously reported methods. Additionally, an assay was developed to confirm the presence of a beta-glycosidic linkage in the purified reaction product and we propose a method by which 3-HKG can be used as a general probe of beta-glucosidase activity. The assay consists of adding glucose oxidase to the 3-HKG/glucosidase solution and then allowing the hydrogen peroxide, generated from the interaction of glucose with glucose oxidase, to oxidize 3-hydroxykynurenine to xanthomattin (XAN) and 4,6 dihydroxyquinolinequinone carboxylic acid (DHQCA). Both XAN and DHQCA absorb strongly between 400 and 500 nm and the color change of the solution can be seen by eye. In addition, XAN fluoresces in the visible region with lambda(max) = 527 nm. PMID- 11726188 TI - In situ deprotection: a method for covalent immobilization of peptides with well defined orientation for use in solid phase immunoassays such as enzyme-linked immunosorbent assay. AB - The orientation of an immobilized antigen is important for recognition by, e.g., an antibody. When noncovalent passive adsorption is used for immobilization, the number of ways that the antigen can attach to the surface is numerous and control of how the antigen orientates on the surface is limited. Covalent immobilization restricts the number of the ways that the antigen can be immobilized to the number of reactive groups on the antigen and, hence, the orientation of the immobilized antigen is more predictable. Peptide antigens were synthesized and purified with protection groups on the lysine and cysteine side chains. These peptides, which have only one good nucleophilic group (the N-terminal alpha-amino group), were immobilized covalently in microtiter plates supplied with tresyl groups on the surface and the protection groups were cleaved off in situ after immobilization. The controlled orientation of these peptides resulted in enhanced recognition by antibodies in general. An enzyme-linked immunosorbent assay for detection of antibodies against a peptide derived from outer surface protein C from Borrelia burgdorferi, found in Lyme borreliosis patients, was established using this strategy. Lyme borreliosis suspect patient sera showed up to a 10-fold increase in the signal when the orientation of the peptide antigen was controlled by the in situ deprotection strategy. PMID- 11726189 TI - Development of a time-resolved fluorescence resonance energy transfer assay (cell TR-FRET) for protein detection on intact cells. AB - An assay named Cell TR-FRET based on time-resolved fluorescence resonance energy transfer, here utilized for detection of receptor proteins on intact cells, is described. In this assay, intact membrane-biotinylated Sf9 cells expressing human interleukin-2Ralpha due to infection with a recombinant baculovirus were prelabeled with a streptavidin-europium (Eu(3+)) chelate, the donor. These prelabeled cells were used in a homogeneous assay by addition of a fluorochrome labeled anti-hIL-2Ralpha-specific antibody, 7G7B6-Cy5, the acceptor. Binding of 7G7B6-Cy5 to hIL-2Ralpha expressed on the cell surface and europium-labeled streptavidin to surface biotin esters brings the donor and the acceptor in close proximity, allowing transfer of energy from the excited state donor to the acceptor. This energy transfer was specifically inhibited by unlabeled antibody and by free biotin. The described assay constitutes a general method since no specific component of the cell membrane is labeled, thereby allowing a number of binding studies on the cell membrane, including receptor density determinations, to be performed. In addition, due to the rapid fashion in which the Cell TR-FRET assay is accomplished, it can be a valuable method not only for identifying novel membrane-associated proteins, but also for drug screening of large samples in high-throughput format. PMID- 11726191 TI - Defining the water content in oriented lipid films by Karl-Fischer titration. PMID- 11726190 TI - A new data analysis method to determine binding constants of small molecules to proteins using equilibrium analytical ultracentrifugation with absorption optics. AB - In principle, equilibrium analytical ultracentrifugation (AU) can be used to quantify the binding stoichiometry and affinity between small-molecule ligands and proteins in aqueous solution. We show here that heteromeric binding constants can be determined using a data-fitting procedure which utilizes a postfitting computation of the total amount of each component in the centrifuge cell. The method avoids overconstraining the fitting of the radial concentration profiles, but still permits unique binding constants to be determined using measurements at a single wavelength. The computational program is demonstrated by applying it to data obtained with mixtures of a 500-Da molecule and interleukin-2, a 16-kDa protein. The 1:1 binding stoichiometry and heteromeric dissociation constants (K(ab)) determined from centrifuge data at two different wavelengths are within the 4-9 microM range independently determined from a functional assay. Values for K(ab) have been obtained for ligands with affinities as weak as 500 microM. This AU method is applicable to compounds with significant UV absorbance (approximately 0.2) at concentrations within approximately 5- to 10-fold of their K(ab). The method, which has been incorporated into a user procedure for IgorPro (Wavemetrics, Oswego, OR), is included as supplementary material. PMID- 11726192 TI - Detection of ATP competitive protein kinase inhibition by Western blotting. PMID- 11726193 TI - Rapid measurement of low levels of sodium, potassium-ATPase activity by ascorbic acid reduction without strong acid. PMID- 11726194 TI - Detecting genotypes of catalase mutant mice by genomic polymerase chain reaction and restriction analysis. PMID- 11726195 TI - Catalysis of guanine nucleotide exchange on eIF2 by eIF2B: can it be both a substituted enzyme and a sequential mechanism? AB - There are conflicting reports over the question of whether the displacement by GTP of GDP bound to eIF2 catalyzed by eIF2B follows a substituted enzyme mechanism, as is believed to be the case for other guanine nucleotide exchange factors, or is a sequential mechanism. Analysis of data recently provided by Williams et al. (J. Biol. Chem. 276, 24697-24703, 2001) showing displacement by eIF2B of GDP bound to eIF2 in the absence of displacing nucleotide appears to offer a way of resolving the dispute and suggests that both mechanisms may be operative. PMID- 11726196 TI - Real time RT-PCR shows correlation between retinoid-induced apoptosis and NGF-R mRNA levels. AB - Neurotrophins and retinoic acid have a critical role in the differentiation and the survival of neurons. All-trans-, 9-cis-retinoic acid (10(-6) M) or NGF (50 100 ng/ml) induced morphologic differentiation and inhibited cell growth in SH SY5Y neuroblastoma cells after 7 days of culture. Continuous treatment of undifferentiated cells with all-trans- or 9-cis-retinoic (10(-6) M) did not induce apoptosis, whereas NGF-differentiated cells showed dramatic apoptosis after 2 to 4 days of retinoic acid treatment as evidenced by TUNEL reaction and flow cytometry analysis following propidium iodide staining. Addition of Ro41 5253 blocked all-trans-retinoic-induced apoptosis, suggesting that the apoptotic signaling pathway was mediated by RARs. The effects of all-trans- or 9-cis retinoic acid on the expression of NGF receptors was evaluated using real-time fluorescence reverse transcription-PCR. A slight transient increase in the expression of p75(NGFR) mRNA was observed by 2 to 4 h after retinoid treatment of undifferentiated cells, whereas a larger increase in the expression of both TrkA and p75(NGFR) mRNA up to threefold the basal level, was observed by 2 to 6 h after retinoid treatment of NGF-differentiated cells. Our results suggest that NGF-differentiated cells may be more susceptible to retinoid-induced apoptosis than undifferentiated cells. PMID- 11726197 TI - Structure-function studies of linear and cyclized peptide antagonists of the GnRH receptor. AB - Structurally new analogs of the peptidic GnRH receptor antagonist Cetrorelix as well as conformationally constrained cyclized deca- or pentapeptides were synthesized and selected peptides evaluated comprehensively. To understand how structural variations of the antagonistic peptide effect pharmacodynamic properties, binding affinities and antagonistic potencies toward the human and rat GnRH receptor were determined. Whereas large substituents in position 6 of linear peptides are compatible with high binding affinity (K(D) < 0.5 nM), all cyclized peptides except the cyclo[3-10] analog D-52391 depicted low binding affinity (K(D) > 10 nM). Binding affinity and antagonistic potency in vitro correlated for all peptides and surprisingly no discrimination between human and rat receptor proteins was observed. Since receptor residues W(101) and N(102) are involved in agonist and antagonist binding, equally potent but structurally different antagonists were tested for binding to the respective W(101)A and N(102)A mutants. In contrast to linear decapeptides, residues N(102) and W(101) are not involved in binding of D-23938 and W(101) is the critical residue for D 52391 binding. We conclude that although equally potent, peptidic GnRH receptor antagonists do have distinct interactions within the ligand binding pocket. Finally, selected antagonists were tested for testosterone suppression in male rats. The duration of testosterone suppression below castration levels differed largely from 1 day for Ganirelix to 27 days for D-23487. Systemic availability became evident as the most important parameter for in vivo efficacy. PMID- 11726198 TI - Restored vulnerability of cultured endothelial cells to high glucose by iron replenishment. AB - High glucose (HG) concentrations are toxic to various cells in vivo, but cells become insensitive to HG toxicity when they are subcultured serially in vitro. Oxidative stress is involved in HG toxicity, and metal ions, especially iron, mediate some oxidative stress. To investigate mechanisms involved in the insensitiveness of cultured cells to HG toxicity, we focused on the level of intracellular iron. Freshly prepared human umbilical vein endothelial cells contained a substantial amount of iron, whereas its level decreased rapidly during the course of culture (to less than 10%). The iron content was restored by incubation of the cells with Fe(III)/8-hydroxyquinoline, and the iron supplemented cells were more susceptible to both oxidant- and HG-induced injury. Under the HG conditions, the iron-loaded cells were subjected to higher levels of oxidative stress. The enhanced HG toxicity by iron was attenuated by the treatment with several antioxidants including catalase, ascorbic acid, and pyruvate. These data suggested that the insensitiveness of subcultured cells to HG toxicity is, at least in part, due to rapid and dramatic loss of intracellular iron. Supplementation with iron is useful to restore the vulnerability of cultured cells to HG that is normally observed in in vivo situations. PMID- 11726199 TI - LIGHT, a member of the TNF superfamily, induces morphological changes and delays proliferation in the human rhabdomyosarcoma cell line RD. AB - LIGHT is a member of the tumor necrosis factor (TNF) superfamily, which binds two known receptors, lymphotoxin-beta receptor (LTbetaR) and the herpesvirus entry mediator (HVEM)/TR2. We investigated the effects of LIGHT on the human rhabdmyosarcoma cell line RD. LIGHT delayed cell proliferation and induced morphological changes of the cells. These effects were not shown by other TNF family ligands such as TNFalpha and LTalpha, which induced the transcriptional activity of nuclear factor-kappaB (NF-kappaB) and NF-kappaB-responsible chemokine productions in the same manner as did LIGHT. LTalpha1beta2, another TNF family ligand for LTbetaR, was shown to have similar activities in RD cells as LIGHT. Both LIGHT and LTalpha1beta2 induced the expression of muscle-specific genes such as smooth muscle (SM) alpha-actin, while TNFalpha and LTalpha did not. These findings indicate that LIGHT may be a novel inducer of RD cell differentiation associated with SM alpha-actin expression through the LTbetaR. PMID- 11726200 TI - Wild-type and mutated nicastrins do not display aminopeptidase M- and B-like activities. AB - Nicastrin is a recently discovered protein interacting with presenilins and the beta-amyloid precursor protein, the proteins playing key roles in Alzheimer's disease and which, when mutated, appear responsible for early-onset familial forms of Alzheimer's disease. Nicastrin was reported to modulate beta-amyloid production, a phenotype affected differently by missense mutations or deletions of a conserved hydrophilic domain. In addition to such a function, nicastrin was recently suggested to possess putative catalytic activity based on its sequence homology with enzymes of the aminopeptidase family. We set up stably transfected human HEK293 cells expressing either wild-type or mutated nicastrins and we show that these proteins do not exhibit aminopeptidase M- and B-like activities. PMID- 11726201 TI - DNA methylation of the human oxytocin receptor gene promoter regulates tissue specific gene suppression. AB - In the human oxytocin receptor (OTR) gene, there is a CpG island from 140 bp upstream to 2338 bp downstream of the transcription start site (TSS). We investigated whether the methylation state of this region affects the transcription of the OTR gene. HepG2 derived from human hepatoblastoma, in which OTR gene transcription was suppressed, was treated with a demethylating agent, 5 azacytidine (Aza-C) for 2 days. Semiquantitative RT-PCR indicated that OTR mRNA was significantly increased by Aza-C treatment in a dose-dependent manner. We estimated the level of methylation within the CpG islands of the OTR gene in peripheral blood leukocytes, nonpregnant uterine myometrium, term uterine myometrium and liver. A 1.5-kb region located 5' upstream of the translation start site was divided into four fragments. Each was amplified by PCR after complete digestion with methylation-sensitive restriction enzyme HpaII. The amount of PCR products was largest in the liver, suggesting that this CpG island in the OTR gene is most highly methylated in liver, where the gene is always inactivated. We compared the effect of in vivo methylation of the CpG island on transcriptional activity of an OTR-reporter plasmid. The reporter gene activity of expression plasmid -2860/+1342-GL3, containing the CpG island, in HepG2 cells was suppressed to 30.6% of the control level after methylation with SssI methylase, while that of -2840/+144-GL3, without the CpG island was suppressed only to 81.4%. The deletion of the segment (MT2) where the level of methylation was most different between liver and uterus (-2860/+1342(del)MT2-GL3) rescued the suppression rate to 68.0%. These results indicate that the methylation of the CpG island in the human OTR gene promoter suppressed its transcription at least in liver and may regulate tissue specific gene expression among organs. PMID- 11726202 TI - Chromium chloride inhibits oxidative stress and TNF-alpha secretion caused by exposure to high glucose in cultured U937 monocytes. AB - Chromium supplementation has been proposed to promote the action of insulin and the lowering of blood glucose levels in diabetic patients. However, the molecular mechanism by which chromium increases insulin sensitivity is not known. Using U937 monocytes as a cell culture model, this study demonstrates that chromium inhibits the secretion of TNF-alpha, a cytokine known to inhibit the sensitivity and action of insulin. U937 cells were cultured with high levels of glucose (mimicking diabetes) in the presence or absence of chromium chloride in the medium at 37 degrees C for 24 h. This study demonstrates that chromium supplementation prevents the increase in TNF-alpha levels and oxidative stress caused by the high levels of glucose in cultured U937 monocytic cells. Similarly, chromium supplementation prevented elevated TNF-alpha secretion and lipid peroxidation levels in H(2)O(2)-treated U937 cells. This study demonstrates for the first time that chromium supplementation inhibits TNF-alpha secretion in U937 monocytes cultured in high-glucose medium, which appears to be mediated by its antioxidative effect. This provides evidence for a novel molecular mechanism by which chromium supplementation may increase insulin sensitivity and glycemic control in diabetic patients. PMID- 11726203 TI - Differential expression of estrogen receptor beta (ERbeta) and its C-terminal truncated splice variant ERbetacx as prognostic predictors in human prostatic cancer. AB - Estrogens have been widely used for the treatment of advanced prostatic adenocarcinoma. However, their direct effect to prostatic cancer cells via estrogen receptors remains unclear. We investigated expression of ERalpha, wild type ERbeta (wtERbeta), and a C-terminal truncated splice variant of ERbeta (ERbetacx) in 50 benign and 100 malignant human prostatic tissue samples by immunohistochemistry. While strong immunostaining of ERalpha was consistently identified in the stromal compartment, wtERbeta was expressed in epithelial cells in both the benign and malignant foci. However, wtERbeta expression was significantly lower in the cancers than in the benign epithelium and inversely correlated with Gleason tumor grade (P < 0.0001 and P = 0.0099, respectively). In contrast, ERbetacx was significantly more expressed in the high-grade cancers (83%) compared with the low-grade tumors (22%) and the benign sites (11%) (P < 0.0001, both). Cancer-specific survival of patients with lower wtERbeta expression was significantly worse than those with higher expression of wtERbeta (P = 0.0018). Conversely, higher ERbetacx expression significantly correlated with poor cancer-specific survival (P = 0.0058). These results suggest that differential expressions of wtERbeta and ERbetacx may be prognostic predictors for prostatic cancer. PMID- 11726204 TI - Conserved negatively charged residues are not required for ATP action at P2X(1) receptors. AB - The role of conserved negatively charged aspartic (D) and glutamic (E) acid residues within the ectodomain of the human P2X(1) receptor were examined by alanine substitution mutagenesis. Effects on ATP potency and cell surface localisation were assessed in Xenopus oocytes using the two electrode voltage clamp technique and cell surface biotinylation. Of the eleven residues tested no major shifts in ATP potency were observed with EC(50) values for ATP ranging from 0.8 to 4.3 microM (compared to 1 microM ATP for wild-type P2X(1) receptors). Peak current amplitudes for mutants D86A and D264A where reduced by approximately 90% due to a corresponding reduction in both total protein and cell surface expression. These results demonstrate that individual conserved negatively charged amino acids are not essential for ATP recognition by the human P2X(1) receptor and coordinated binding of the positive charge on magnesium complexed ATP by negatively charged amino acids is not required. PMID- 11726205 TI - Tamoxifen agonism and estrogen antagonism of c-fos gene promoter activity through non-consensus-responsive elements in MC3T3-E1 osteoblasts. AB - We find that the activity of a 0.4-kb human c-fos gene promoter (-404/+41), which lacks consensus estrogen-responsive elements (EREs), is regulated by estrogen receptor (ER) ligands in MC3T3-E1 osteoblastic cells through ERs in a manner distinct from ERE-mediated regulation. When ERalpha is coexpressed, both estrogens and antiestrogens upregulate promoter activity. When ERbeta is coexpressed, however, three tested antiestrogens affect c-fos promoter activity, with tamoxifen exerting the greatest effect, while estrogens have no such effect. The tamoxifen agonism through ERbeta is antagonized by 17beta-estradiol, while the 17beta-estradiol agonism through ERalpha is canceled by excess-level coexpression of ERbeta. Deletion analysis revealed that the sequence -206/-110 plays a crucial role in the ERbeta-mediated tamoxifen agonism. Interestingly, there is no ERbeta-mediated tamoxifen agonism when nonosteoblastic cells are tested. Taken together, these results suggest that the transcription of the c-fos gene is regulated by ER ligands possibly through non-ERE elements in ligand structure-, cell type-, and ER subtype-dependent manners. PMID- 11726206 TI - Structural role of conserved Asn179 in the short-chain dehydrogenase/reductase scaffold. AB - Short-chain dehydrogenases/reductases (SDR) constitute a large family of enzymes found in all forms of life. Despite a low level of sequence identity, the three dimensional structures determined display a nearly superimposable alpha/beta folding pattern. We identified a conserved asparagine residue located within strand betaF and analyzed its role in the short-chain dehydrogenase/reductase architecture. Mutagenetic replacement of Asn179 by Ala in bacterial 3beta/17beta hydroxysteroid dehydrogenase yields a folded, but enzymatically inactive enzyme, which is significantly more resistant to denaturation by guanidinium hydrochloride. Crystallographic analysis of the wild-type enzyme at 1.2-A resolution reveals a hydrogen bonding network, including a buried and well ordered water molecule connecting strands betaE to betaF, a common feature found in 16 of 21 known three-dimensional structures of the family. Based on these results, we hypothesize that in mammalian 11beta-hydroxysteroid dehydrogenase the essential Asn-linked glycosylation site, which corresponds to the conserved segment, displays similar structural features and has a central role to maintain the SDR scaffold. PMID- 11726207 TI - Activation of JNK and transcriptional repressor ATF3/LRF1 through the IRE1/TRAF2 pathway is implicated in human vascular endothelial cell death by homocysteine. AB - Endothelial cell injury underlies an increased occurrence of thromboembolic vascular disease in hereditary hyperhomocysteinemia. We have previously shown that homocysteine causes activation of c-Jun NH(2)-terminal kinase (JNK) and activating transcription factor 3/liver regenerating factor 1 (ATF3/LRF1) and induces apoptosis in human umbilical vein endothelial cells (HUVECs). In this study, the activation of JNK and ATF3 in HUVECs was mediated by the endoplasmic reticulum (ER) resident transmembrane kinase IRE1alpha and beta, which sense and transduce signal of the accumulationj of unfolded proteins in the ER. Moreover, dominant negative mutants of tumor necrosis factor receptor-associated factor 2 and mitogen-activated kinase kinase 4 and 7, as well as antisense ATF3 cDNA, inhibited cell death by homocysteine. These results indicate that the activation of JNK and ATF3 through the ER stress of homocysteine plays a role in the homocysteine-induced cell death. The JNK-ATF3 pathway may be implicated in endothelial cell injury associated with hereditary hyperhomocysteinemia. PMID- 11726208 TI - Growth factor regulation of the molecular chaperone calnexin. AB - Heregulin-beta1 (HRG) is a regulatory polypeptide having several distinct biological effects in mammary epithelial cells. To address the hypothesis that HRG selectively regulates gene expression, we performed differential display screening using cells grown in the presence or absence of HRG. One cDNA clone upregulated by HRG was identical to human calnexin, a protein with molecular chaperone function. This is the first demonstration of the regulation of calnexin mRNA and protein expression by a physiologically relevant polypeptide factor in human breast cancer cells. HRG stimulation also caused a rapid redistribution of calnexin from vesicle-like structures in the cell cytoplasm to the perinuclear area and to the cell membrane. Furthermore, HRG induced colocalization and physical interaction of calnexin with the HER2 growth factor receptor. Finally, calnexin protein levels were increased in progressive stages of human breast cancer. These findings suggest that stimulation of calnexin expression by HRG may constitute a mechanism of protein redistribution and facilitate downstream signaling events in growth-factor-activated cells. PMID- 11726209 TI - Analysis of chromatin-immunopurified MeCP2-associated fragments. AB - As molecular biologists, we are continuing to unravel the interactions by which DNA binding proteins mediate the expression of genes. The chromatin immunoprecipitation (ChIP) technique provides us with an exquisite tool to investigate the interplay between chromatin structure and its role in regulating transcription, replication, and recombination in vivo. We describe a robust assay used to identify the molecular determinants associated with chromatin. In this article we illustrate the ChIP technique and use the transcriptionally silent hypermethylated multidrug resistance (MDR1) gene as the platform for methyl-CpG binding protein 2 (MeCP2) localization on chromatin. Driven by the hypothesis that repression is strongly dependent on the methylation profile of the endogenous promoter, we demonstrate that MDR1 is targeted by MeCP2. Methylated MDR1 chromatin is highly enriched with MeCP2 and is in striking contrast to localization observed in cells in which MDR1 is transcriptionally active. In a distinct model system we discuss experimental methods used to immunopurify the MeCP2 repressor complex on chromatin and quantify protein-DNA association by competitive PCR approach. PMID- 11726210 TI - Identification of structural domains affecting transactivation potential of Nm23. AB - The strong transactivation activity of the C-terminal half (amino acids 76-152) of Nm23 was reported previously. Here we examined a structural domain preventing or necessary to its transactivation activity. The C-terminal 1/4 (amino acids 109 152) was sufficient for transactivation, but the C-terminal half with a longer N terminal extension (amino acids 58-152) caused the loss of the transactivation ability. Furthermore, coexpression of the N-terminal half with the C-terminus of Nm23-H1 blocked the transactivation activity of the C-terminal half, where direct interaction of both truncated proteins was demonstrated in vitro. Transactivation activities in the C-terminal halves of the known mutants (P96S, H118F, S120G, and S120A) exhibiting differential antimetastasis effects were also tested. Significant reduction of transactivation activity was observed only in H118F, indicating that NPD kinase active-site histidine is required. This suggests that transactivation potential of Nm23 is related to NDP kinase activity but not to metastasis suppressor activity. PMID- 11726211 TI - Antiproliferative and antimitogenic activities in a peptide from puffball mushroom Calvatia caelata. AB - A peptide with a molecular weight of 8 kDa and an N-terminal sequence closely resembling that of ubiquitin was isolated from fruiting bodies of the mosaic puffball mushroom Calvatia caelata. The peptide was purified using a protocol that involved ion-exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, and ion-exchange chromatography on Mono S. The peptide inhibited translation in the cell-free rabbit reticulocyte lysate system and exhibited N-glycosidase activity. It potently inhibited proliferation of spleen cells with an IC(50) of about 100 nM as indicated by the suppression of [methyl-(3)H]thymidine uptake. The viability of breast cancer cells was reduced to half at a ubiquitin concentration of about 100 nM. PMID- 11726212 TI - Pleureryn, a novel protease from fresh fruiting bodies of the edible mushroom Pleurotus eryngii. AB - A protease designated pleureryn, with an N-terminal sequence dissimilar from previously reported mushroom metalloendopeptidases and showing only limited resemblance to aspartic proteinases, albeit considerable homology to DNA replication licensing factor, was isolated from fresh fruiting bodies of the edible mushroom Pleurotus eryngii. The purification protocol entailed ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel Blue gel, ion exchange chromatography on CM-Sepharose, and FPLC-gel filtration on Superdex 75. The protease was unadsorbed on DEAE-cellulose but adsorbed on Affi gel Blue gel and CM-Sepharose. It demonstrated a single band with a molecular weight of 11.5 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Pleureryn demonstrated a protease activity of 9364 U/mg toward casein. It exhibited a pH optimum of 5.0 and a temperature optimum of 45 degrees C, with substantial activity remaining at high temperatures and pH 4 and 12. The activity of the protease was adversely affected by pepstatin A, indicating that it is an aspartic protease. PMSF, trypsin inhibitor, and EDTA exerted no striking effect, suggesting that it is neither a serine protease nor a metalloprotease. It inhibited translation in a rabbit reticulocyte lysate system with an IC(50) of 20 nM. Pleureryn also exhibited some inhibitory activity against HIV-1 reverse transcriptase, reminiscent of a suppressive action of HIV-1 protease on its homologous reverse transcriptase but was devoid of ribonuclease, deoxyribonuclease, and antifungal activities. PMID- 11726213 TI - Hypermutable change of human UV(r)-1 cells by p53 overexpression. AB - The p53 protein has been reported to regulate cellular responses to genetic stress such as far-ultraviolet light (UV), protecting human cells from mutation. Levels of p53 protein in hypermutable RSa cells were found here to increase soon after UV irradiation, while those in UV(r)-1 cells, a hypomutable variant of RSa cells, showed a delayed increase. Three cell lines overexpressing wild-type p53 in UV(r)-1 cells exhibited higher sensitivity to UV mutagenicity than did control U-V-7 cells transfected with vector alone, assessed using the ouabain-resistance phenotypic mutation test and identification of K-ras codon 12 base substitution mutation. On the other hand, U-V-7 cells showed UV-induced elevation of antipain sensitive protease activity, but p53 transfectants did not. Moreover, antipain treatment to U-V-7 cells was increased susceptibility to UV mutagenicity. Thus, p53 protein overproduction may sensitize human cells, at least those tested, to UV mutagenicity, in association with inhibition of protease activity. PMID- 11726214 TI - N-terminal activation function is dominant in ligand-dependent transactivation of medaka estrogen receptor alpha in human cells. AB - Two independent transcriptional activation functions have been mapped to the N- and C-terminal domains of estrogen receptors (ERs), and are named activation function-1 (AF-1) and AF-2, respectively. Due to the lower activity of AF-1 and difficulties in producing AF-1 recombinant protein, little information is available regarding the biochemical properties of ER AF-1 and its coactivators compared to AF-2. In this study, we characterized the AF domains from medaka fish ERalpha (meERalpha) using a transient expression assay in cultured mammalian cells. While both meERalpha AF-1 and AF-2 were functional and gave similar results to human ERalpha AFs, meERalpha AF-1 displayed significant activity even in HeLa cells that exhibit little human ERalpha (hERalpha) AF-1 activity. Evidence of transcriptional squelching between hERalpha and meERalpha AF-1 molecules suggested that the molecules utilized common coactivators in mammalian cells. We also showed that large amounts of the meERalpha A/B domain could be expressed in Escherichia coli cells as a soluble protein, in contrast to hERalpha A/B domain protein which was not observed. Taken together, our results suggested that meERalpha AF-1 may have a more significant role in estrogen-induced function of meERalpha than AF-2 in medaka fish. PMID- 11726215 TI - Hysteresis and positive cooperativity of iceberg lettuce polyphenol oxidase. AB - A kinetic study of the diphenolase activity of latent polyphenol oxidase (PPO), purified from Iceberg lettuce (Lactuca sativa L), revealed a sigmoid relationship between the reaction rate and the substrate concentration with a high Hill coefficient (n(H) = 3.8). This positive cooperativity had not been previously described for any PPO. Furthermore, the enzyme showed a lag phase in the expression of this activity, suggesting a hysteretic nature of the enzyme. The kinetic behavior, the latency and the lag phase varied at different steps of the purification process. PPO showed hyperbolic or cooperative kinetics depending on the pH assay and the sodium dodecyl sulfate (SDS) concentration. Substrate induced slow conformational change of the oligomeric enzyme is suggested. The conformational change would be toward a more active enzyme form with higher affinity for the substrate and favoured by acid pH and SDS. PMID- 11726216 TI - Use of muscarinic agonists in the treatment of Sjogren's syndrome. AB - Two muscarinic agonists (pilocarpine and cevimeline) have recently been approved for the treatment of symptoms of xerostomia in Sjogren's syndrome (SS). These agents stimulate the M1 and M3 receptors present on salivary glands, leading to increased secretory function. The use of these agents emphasizes the importance of neuroendocrine mechanisms in SS, which is considered an autoimmune disorder. We review recent studies on the release of cytokines and metalloproteinases in SS affected glands and their influence on the release of and response to neurotransmitters. Also, we review the structure and function of muscarinic receptors as they may relate to SS and the potential use of novel muscarinic agonists in SS. PMID- 11726217 TI - Expression of inhibitory receptors in natural killer (CD3(-)CD56(+)) cells and CD3(+)CD56(+) cells in the peripheral blood lymphocytes and tumor infiltrating lymphocytes in patients with primary hepatocellular carcinoma. AB - The cytolytic responses of NK (CD3(-)CD56(+)) and CD3(+)CD56(+) cells are inhibited by the engagement of the killer inhibitory receptors (p58.1, p58.2, and CD94) with respective ligands on the target cell. The expression of these receptors in peripheral blood lymphocytes (PBLs) (n = 18) and tumor infiltrating lymphocytes (TILs) (n = 7) was examined in patients with primary hepatocellular carcinoma (HCC). There were no differences in the expression of the three inhibitory receptors by both NK and CD3(+)CD56(+) PBLs in patients with HCC compared to that of control NK and CD3(+)CD56(+) PBLs, respectively (all P = NS). However, the expression of p58.1 by NK TILs and by CD3(+)CD56(+) TILs in patients with HCC was significantly decreased compared to that of hepatic lymphocytes of the control subjects (8.9% vs 37.85%, P = 0.047; 4.1% vs 25.2%, P = 0.049, respectively). The expression of p58.2 by CD3(+)CD56(+) TILs and CD94 by NK TILs was also decreased compared to that of hepatic lymphocytes of the control subjects (16.9% vs 73.1%, P = 0.047; 21% vs 49.95%, P = 0.037, respectively). These changes were limited to hepatic TILs, and this observation may reflect an adaptive anti-tumor phenomenon occurring in the microenvironment of HCC. PMID- 11726218 TI - Analysis of SWAP-70 as a candidate gene for non-X-linked hyper IgM syndrome and common variable immunodeficiency. AB - SWAP-70 is a recently identified protein that functions as the only B cell specific component of an isotype switch recombination complex called SWAP. The SWAP complex has specificity for the switch regions upstream of the constant region immunoglobulin genes and it facilitates the transfer of DNA between switch regions. These features suggested that mutations in the gene encoding SWAP-70 might result in humoral immunodeficiency. To test this hypothesis we determined the genomic structure of this gene and used single-stranded conformational polymorphism (SSCP) analysis to screen DNA from 38 patients with either non-X linked hyper IgM syndrome or common variable immunodeficiency. The results demonstrated that SWAP-70 consists of 12 exons spread over 89 kb at chromosome 11p15.2. SSCP analysis of the patient population revealed five polymorphic variants in the gene, one of which (Q505E) is an amino acid substitution in the putative nuclear export signal of SWAP-70. However, none of the alterations appeared to be associated with disease in the patients screened. PMID- 11726220 TI - Outcome of intravenous immunoglobulin-transmitted hepatitis C virus infection in primary immunodeficiency. AB - Physicians in the United States who treat patients with primary immunodeficiency were contacted to identify subjects who had been infected with hepatitis C due to exposure to contaminated intravenous immunoglobulin (IVIg) in 1993-1994. From this survey we gathered information on 58 PCR-positive hepatitis C-infected patients; 37 had CVID, 9 had XLA, 5 were IgG subclass deficient, 4 were antibody deficient with normal immunoglobulin levels, 2 had SCID after BMT, and 1 had B cell linker deficiency. Of the 58 subjects, 30 had been treated with IFN-alpha in combination with ribavirin in 5 cases, and 26 other subjects were not treated. Of those who were treated, 11 (37%) resolved the infection and became PCR-negative; of the 26 who were not treated, 5 (19%) have resolved the infection, outcomes not significantly different. Patients 20 years of age or younger had a significantly better outcome compared to those older than age 20 (P = 0.02). Five subjects of the 58 have had a liver transplantation, a sixth has had two transplants, and 10 (17%) of the group have died. This survey demonstrates the heterogeneity of the clinical outcome in subjects with primary immunodeficiency who contracted hepatitis C due to viral contamination of IVIg. PMID- 11726219 TI - Generation of CD4(+) and CD8(+) T lymphocyte responses by dendritic cells armed with PSA/anti-PSA (antigen/antibody) complexes. AB - Dendritic cells (DC) acquire antigens through a number of cell surface structures including receptors for the Fc portion of immunoglobulins and mannose. Little is known about the effects of antigen uptake via these receptors on antigen processing and presentation. We compared the capacity of DC to generate CD4(+) and CD8(+) T cell responses after exposure to prostate-specific antigen (PSA) alone, PSA targeted to the mannose receptor (mannosylated PSA (PSA-m)), or PSA targeted to Fc receptors by combining PSA with an anti-PSA antibody (AR47.47). Autologous CD3(+) T cells were added to monocyte-derived immature DC that had been cultured with GM-CSF/IL-4 for 4 days, exposed to antigen, and matured with CD40L or TNFalpha/IFN-alpha. After several rounds of stimulation, T cell responses were assessed by intracellular IFN-gamma production using flow cytometry. Both CD4(+) and CD8(+) T cell responses were observed after stimulation with DC exposed to the PSA/anti-PSA complexes, whereas CD4(+) predominated over CD8(+) T cell responses after stimulation with PSA-armed DC or PSA-m. These CD8(+) T cells responded when rechallenged with DC pulsed with HLA allele-restricted PSA peptides. These results indicate that PSA and PSA-m are processed primarily through pathways that favor HLA Class II presentation, while the PSA/anti-PSA immune complexes are processed through both Class I and Class II pathways in monocyte-derived DC. These findings have potential applications in designing more effective cancer vaccines for prostate cancer. PMID- 11726221 TI - Cellular immune response against tropomyosin isoform 5 in ulcerative colitis. AB - We have reported an autoantibody response in ulcerative colitis (UC) against human tropomyosin isoform 5 (hTM5), the predominant colonic epithelial cell hTM isoform. In this report, we determined the number of IFN-gamma-secreting cells (spot-forming cells, SFC) against hTM5 by an enzyme-linked immunospot (ELISPOT) assay. Another cytoskeletal protein, caldesmon, CaD40, was used as a control antigen. Peripheral blood mononuclear cells were separated by a Ficoll density gradient from 28 patients with UC, 13 patients with Crohn's disease (CD), and 9 healthy subjects (HS). The mean (+/-SEM) SFC values against hTM5 in UC, CD, and HS were 48.8 +/- 8.1, 18.6 +/- 4.6, and 20.8 +/- 8.6, respectively. The value in UC was significantly higher than those in CD (P < 0.005) and HS (P < 0.025). SFC values in CD did not differ from those in HS. None of the 50 samples (except 1 UC) reacted to the CaD40 antigen. This study demonstrates, for the first time, a defined colon epithelial cell antigen, hTM5, that is capable of inducing a significant T cell response in UC but not in CD. PMID- 11726222 TI - Identification of peptide sequences specific for serum antibodies from human papillomavirus-infected patients using phage display libraries. AB - Three random phage display peptide libraries were screened with sera from human papillomavirus (HPV)-infected patients to characterize the specificities of antibodies present in patients' sera and to identify molecules that correspond to or mimic natural epitopes; 141 phage clones were randomly selected in three rounds of bioselection and their binding properties were analyzed in ELISA using sera from 36 patients with confirmed HPV 16 infection and 24 healthy control female blood donors. Sixteen of 36 (44%) patients' sera reacted with at least 1 phage clone, and only 2 of 24 female donors' sera showed positive reaction with 1 of the selected clones. We conclude that the combination of various disease specific epitopes generated by screening of phage display peptide libraries may potentially lead to a multicomponent diagnostic assay for the early detection of HPV infection and precancerous cervical lesions, making possible the prevention of one of the most common cancers in women. PMID- 11726223 TI - Efficacy and mechanisms of action of rmB7.2-Ig as an antitumor agent in combination with Adriamycin and Cytoxan chemotherapy. AB - The efficacy of chemotherapy for cancer is often limited by toxicity. Immune approaches to cancer immunotherapy, while promising for specificity and long-term protection, have not typically proven potent enough to generate significant therapeutic responses. We have shown therapeutic benefit using recombinant murine B7.2-Ig (rmB7.2-Ig) in murine tumor models. Efficacy was dependent on immune activity and was not associated with toxicity. Recently, the efficacy of rmB7.2 Ig was demonstrated in leukemia tumor models in combination with chemotherapeutic agents. To further explore the potential of this approach, we evaluated the efficacy in solid tumor models of rmB7.2-Ig given in combination with chemotherapeutics commonly used in clinical practice, testing the effects of dose and schedule. RmB7.2-Ig in combination with some chemotherapeutics enhances the activity and efficacy of reduced chemotherapeutic doses. However, the relative timing of chemotherapy and rmB7.2-Ig dosing can be important. Investigation of mechanisms of action based on histological studies suggests that inflammatory as well as T cell mechanisms comprise the response. Additional studies of mice deleted of B7.1, B7.2, and CTLA-4 suggest that the enhanced response induced by rmB7.2-Ig may not be mediated through CD28 ligation alone. The efficacy suggests potential for recombinant human B7.2-Ig as an adjuvant to chemotherapy in promoting immune-mediated mechanisms to augment the activity of chemotherapy. PMID- 11726224 TI - Predominant IL-10 production in indirect alloreactivity is not associated with rejection. AB - In a prospective study of indirect alloresponse in renal transplantation, we detected proliferation and cytokine production to donor and third-party HLA-DR peptides unrelated to rejection. Twenty of 28 patients (71%) presented proliferation, 29% before and 71% after transplantation. Half of the patients also presented proliferation to third-party peptides. Indirect alloresponse was also detected in 75% of healthy individuals (HI). Variability of response was observed in patients and HI for both proliferation and cytokine production. IL-10 predominance was observed in indirect alloresponses to donor peptides pre- and post-Tx, in contrast with more IFN-gamma and TGF-beta being detected in HI. IL-10 production was frequently detected without proliferation, in contrast with more frequent proliferation being found with IFN-gamma and TGF-beta production. The lack of association of either cytokine or proliferation with rejection, together with the predominance of IL-10 unrelated to proliferation, suggests that regulatory cells may be part of the T cell repertoire involved in indirect alloreactivity. PMID- 11726225 TI - CD200 immunoadhesin suppresses collagen-induced arthritis in mice. AB - DBA/1 mice immunized with 100 microg bovine collagen type II emulsified in Freund's adjuvant, followed by booster injection in incomplete adjuvant at 18 days, develop profound arthritis (>50% of animals) by 30 days postinjection. The molecule CD200 (previously called OX2), associated with, among others, follicular dendritic cells, is implicated in delivery of immunosuppressive signals to the immune system, and an immunoadhesin in which the extracellular domains of CD200 were linked to a mouse IgG2a Fc region has been shown to promote renal allograft survival. DBA/1 mice receiving 15 microg/mouse CD200Fc at 3-day intervals following immunization with collagen did not develop arthritis in this model. Lymphocytes taken from CD200Fc-treated, collagen-immunized mice produced significantly lower levels of TNFalpha and IFN-gamma in culture supernatants after restimulation in vitro with collagen, in contrast to cells taken from control mice treated with PBS or normal mouse Ig. Serum from CD200Fc-treated mice contained less anti-collagen IgG (approximately 50% reduction), with relatively more IgG2b and IgG3, and lower levels of TNFalpha and IFN-gamma, than control mice. These data indicate that this immunoadhesin may have a potent role to play in the regulation of autoimmune disorders. PMID- 11726226 TI - Evidence that oxidative stress-induced apoptosis by menadione involves Fas dependent and Fas-independent pathways. AB - Menadione (vitamin K3) a redox cycling quinone, is a clinically important chemotherapeutic agent. The objective of this study was to clarify the cytotoxic mechanisms by which menadione induces cell death in a lymphoblastoid cell line. Our results show that while the Jun kinase cascade and FasL expression may contribute to cell death at lower drug concentrations, a mitochondrial pathway dominates the cytotoxic effect at higher menadione concentrations. Menadione treatment clearly affected the mitochondrial function of Jurkat T cells by inducing a collapse of the inner transmembrane potential (DeltaPsi(m)) and a decrease in inner membrane mass, which could be completely reversed by N acetylcysteine. Importantly, while a broad range of fmk-derived caspase inhibitors had potent effects on Fas-induced apoptosis, they failed to interfere in menadione cytotoxicity, indicating that menadione-induced cell death is predominantly Fas-independent. In addition, the mitochondrial changes coincided with ATP depletion. The failure in ATP production explains the occurrence of Fas independent death events. PMID- 11726227 TI - Recovery from mouse hepatitis virus infection depends on recruitment of CD8(+) cells rather than activation of intrahepatic CD4(+)alphabeta(-)TCR(inter) or NK-T cells. AB - Mouse hepatitis virus (MHV) provides an excellent animal model for the study of the immunopathological mechanisms involved in hepatic viral diseases. We previously generated an attenuated viral variant, YAC-MHV3, which induces a subclinical disease and recovery within 15 days. In contrast, the L2-MHV3 strain induces the development of a fulminant hepatitis, leading to death within 3 days. In this paper, we document intrahepatic and splenic T cell subpopulations involved in the hepatitis process and viral elimination identified in attenuated or pathogenic MHV3-infected C57BL/6 mice. Percentages of intrahepatic CD4(+) cells decreased in attenuated YAC-MHV3-infected mice, while they increased in mice infected with pathogenic L2-MHV3, compared with uninfected animals. Moreover, in YAC-MHV3-infected mice, the percentages of intrahepatic CD8(+) cells slightly decreased at 24 h pi, then increased until 15 days pi. In contrast, the CD4/CD8 ratios of splenic lymphoid subpopulations increased in the first days of infection and returned to normal values at 15 days pi. Intrahepatic NK1.1(+)alphabeta - TCR(inter) cells decreased in both virally infected groups of mice, while CD4(+)alphabeta - TCR(inter) LFA-1(high) cells increased in L2-MHV3 infected mice, in contrast with what was seen in YAC-MHV3-infected mice. However, these cells became anergic following Con A or PHA stimulation. Ex vivo studies showed that only the intrahepatic CD8(+) cells that were increased in YAC-MHV3 infected mice could be stimulated by lectins. In addition, in vitro viral infections revealed that L2-MHV3 viral infection led to an increase of intrahepatic CD4(+)alphabeta - TCR(inter) cells in the absence of CD8(+) cells only. These results indicate that the attenuated phenotype of the YAC-MHV3 virus is related to two different mechanisms: the first involves no increase of intrahepatic CD4(+)alphabeta - TCR(inter) or NK-T cells, while the second favors the recruitment and activation of CD8(+) cells in liver. The results are discussed in relation to the integrity of intrahepatic immune tolerance mechanisms and immune-mediated viral elimination. PMID- 11726228 TI - Polymorphisms of the ICAM-1 gene are associated with inflammatory bowel disease, regardless of the p-ANCA status. AB - The intercellular adhesion molecule-1 (ICAM-1) is of paramount importance for the initiation and propagation of various inflammatory conditions. An increased frequency of allele R241 of the ICAM-1 gene was previously described in p-ANCA negative as compared to p-ANCA-positive ulcerative colitis and vice versa in Crohn's disease. One hundred sixteen healthy unrelated controls, 121 patients with ulcerative colitis, and 96 patients with Crohn's disease were genotyped for two polymorphisms of the ICAM-1 gene (R/G241, exon 4; and K/E469, exon 6), employing dot-blot hybridization and stratified according to their p-ANCA status. When compared with the control group the frequency of the allele R241 (P = 0.024) and the heterozygous genotype R/G241, P = 0.032) were significantly increased in ulcerative colitis, whereas the homozygous genotype G/G241 was found less frequently (P = 0.022). The heterozygous genotype K/E469 was observed less frequently (P = 0.001 and 0.037, resp.) than the homozygous genotype E/E469, which was more frequent in Crohn's disease and ulcerative colitis (P = 0.002 and 0.012, respectively). Further significant differences concerning the allele or genotype distribution were not observed. After stratification for the p-ANCA status significant differences concerning the frequencies of both the R241 and the E469 alleles were not detected when p-ANCA-positive inflammatory bowel disease and p-ANCA-negative inflammatory bowel disease were compared. Ulcerative colitis and Crohn's disease are associated with polymorphisms of the ICAM-1 gene, which might therefore represent a functional candidate gene. However, the observed associations are independent of the p-ANCA status. PMID- 11726229 TI - CTLA-4 gene polymorphisms in Tunisian patients with Graves' disease. AB - Graves' disease (GD) is an organ-specific autoimmune disorder of multifactorial etiology with a polygenic mode of inheritance. A recent report has demonstrated that there is a linkage and an association between the genetic markers of the CTLA-4 gene on chromosome 2q33 and GD. In order to confirm this association in a Tunisian population, three polymorphisms of the CTLA-4 gene were analyzed: the first is at the -318 position from the ATG start codon consisting of a C/T change; the second is in position 49 of exon 1, which lies in the A/G transition; and the third is in the 3' untranslated region with variant lengths of the dinucleotide (AT)n repeat. The genomic DNA from 144 patients with GD and 205 healthy individuals was genotyped after specific polymerase chain reaction amplification. Comparative analysis using a chi(2) test showed a weak yet significant difference in allele frequencies of the A/G dimorphic marker between patients and controls (P < 0.05), and a significant increase of A/A homozygous individuals among patients (21.53 vs 12.7%, P = 0.02, odds ratio (OR) = 1.89) was found. Analyses of CTLA-4 A/G polymorphism with respect to sex showed a significant difference in A/A genotypes between female patients and controls (OR = 2.14; 95%, 1.13 < OR < 4.04, P < 0.05). The distribution of CTLA-4 (AT)n allele frequencies differed between patients and controls (chi(2) = 38.18, 20 degrees of freedom, P = 0.0084) and the highest OR was found with the CTLA-4 (AT)-224-bp allele (OR = 6.43, 1.7 < OR < 28.64; P = 0.001). In conclusion, these results show that the CTLA-4 gene, or one closely associated with it, confers susceptibility to GD in a Tunisian population. PMID- 11726230 TI - Evidence of a role for C4 in modulating interstitial inflammation in experimental glomerulonephritis. AB - Activation of C4 releases into the fluid phase a fragment of the alpha chain, C4a. Unlike the analogous fragments of C3 and C2, there is no evidence for an anaphylatoxic effect of C4a. There is actually some in vitro evidence that it could have a modulating effect on inflammation by inhibiting monocyte chemotaxis. We induced an immune complex glomerulonephritis in wild-type (WT) and C4 knock out (C4KO) mice. Although the glomerular component of the disease did not differ in the two groups of animals, there were marked differences in the accompanying tubulo-interstitial injury. Compared to WT animals, the C4KO mice had significantly more infiltrating interstitial cells (1910 vs 2720/mm(2)), foci of tubular atrophy (6.3 vs 14.8/section), and interstitial space (22 vs 30% of cortex). C4 is expressed constitutively by renal tubular epithelial cells. These data support a role for such local C4 in modulating interstitial inflammation, consistent with in vitro experiments. PMID- 11726231 TI - Identification of a CD8alpha(+) dendritic cell subpopulation in rat spleen and evaluation of its OX-62 expression. AB - Rat spleen DC and bone marrow-derived DC were isolated and characterized by morphology and flow cytometry. We found a CD8alpha(+) DC subpopulation representing 19-48% (27.4 +/- 12.0) of total spleen DC. The OX-62 expression on total spleen DC was 41-59% (51.8 +/- 7.5). Myeloid bone marrow-derived DC were negative for CD8alpha and OX-62. We demonstrated the coexpression of CD8alpha and OX-62 molecules, at least in a portion CD8alpha(+) spleen DC. Both CD8alpha(+) and CD8alpha(-) spleen DC subpopulations separated by MACS were able to induce an in vivo primary immune response to OVA. The immune response induced by the CD8alpha(-) DC subpopulation was higher (P < 0.05). We identified a CD8alpha(+) DC subpopulation in rat spleen less effective in inducing an immune response than CD8alpha(-) DC. Moreover, our results suggest the presence of DC subpopulations with different lineages in DC preparations based on OX-62 expression. PMID- 11726232 TI - A celebration of the scientific contributions of Richard F. Thompson. PMID- 11726233 TI - A personal reflection on knowing Richard Thompson from 1990 to 2000. PMID- 11726234 TI - Depotentiation of vdccLTP requires NMDAR activation. AB - Long-term potentiation is an enduring increase in synaptic efficacy following repeated stimulation of afferent fibers that is thought to underlie memory. In area CA1 of the hippocampus at least two forms of synaptic potentiation coexist at the same synapses; nmdaLTP and vdccLTP. NmdaLTP is induced by Ca2+ entry through NMDARs and is dependent on serine/threonine kinase activation, while vdccLTP is induced through Ca2+ entry through VDCCs and is dependent on tyrosine kinase activation. Depotentiation is a mechanism known to reverse nmdaLTP through phosphatase activation. The depotentiation of vdccLTP has not been previously investigated. We used hippocampal slices (area CA1) from male Long-Evans rats to induce vdccLTP with a 200-Hz tetanus in the presence of 50 microM APV. The 200-Hz tetanus resulted in a slowly developing vdccLTP that remained stable for at least 30 min. Thirty minutes after vdccLTP was induced, a low-frequency tetanus (3, 10, 20, 30, or 40 Hz) was applied in the presence of APV in an attempt to depotentiate vdccLTP. The 3- and 10-Hz low-frequency tetani resulted in no depotentiation. The 20- and 30-Hz tetani partially depotentiated vdccLTP (by approximately 13%), whereas the 40-Hz tetanus resulted in further potentiation. When APV was washed out prior to the 3-Hz low-frequency tetanus, the vdccLTP was completely depotentiated--presumably by NMDAR mechanisms. Our results indicate that vdccLTP is resistant to depotentiation under low-frequency stimulation conditions that readily depotentiate nmdaLTP. As tetanus frequencies are increased a small depotentiation is observed, suggesting that vdccLTP can be depotentiated to a small extent. When NMDARs are unblocked, vdccLTP can be completely depotentiated by a 3-Hz low-frequency tetanus, suggesting that vdccLTP can be depotentiated via activation of NMDAR mechanisms. PMID- 11726236 TI - Comparison of single unit responses to tone, light, and compound conditioned stimuli during rabbit classical eyeblink conditioning. AB - Unit recordings and lesion studies have implicated the cerebellum as an essential site for the acquisition and maintenance of the conditioned eyeblink response. The current study looked at the neural characteristics of conditioned stimulus (CS) processing in the interpositus nucleus of the cerebellum after training New Zealand white rabbits (Oryctolagus cuniculus) in one of two conditioning paradigms: (a) compound conditioning (CMP), a compound CS consisting of light and tone paired with an air puff unconditioned stimulus (US); or (b) stimulus compounding (ALT), alternating blocks of tone CS and light CS trials paired with the air puff US. Single unit responses were recorded during five sessions after the animals had reached an asymptotic level of responding. Animals were tested for behavioral and neural responses to CS alone trials that included tone alone, light alone, and compound tone-light trials. For the CMP group, the compound CS elicited 80 to 90% conditioned eyeblink responses (CRs), whereas the individual tone and light CSs elicited only 40 to 50% CRs. For the ALT group, all three CSs (tone, light, and compound) elicited very high levels of responding of at least 80% CRs. For the CMP group, there were roughly equal numbers of cells responding to all of the CSs. This includes cells that responded exclusively to one, and only one, of the three stimuli and also those cells that responded to combinations of two or more. Cells from the ALT group were far more likely to respond exclusively to only one of the CSs. Both the behavioral and physiological results suggest that the compound tone-light stimulus was processed as a distinct stimulus, separate from the component tone and light. These results are discussed in the context of multisensory processing. PMID- 11726235 TI - 17beta-estradiol: effect on CA1 hippocampal synaptic plasticity. AB - An understanding of synaptic plasticity in the mammalian brain has been one of R. F. Thompson's major pursuits throughout his illustrious career. A current series of experiments of significant interest to R. F. Thompson is an examination of the interactions between sex hormones, synaptic plasticity, aging, and stress. This research is contained within a broader project whose aim is to investigate animal models that evaluate estrogen interactions with Alzheimer's disease. This paper reviews the recent results that have led to a better understanding of how the sex hormone estrogen influences synaptic plasticity in an important structure within the mammalian brain responsible for learning and memory: the hippocampus. In this review, a number of experiments have been highlighted that investigate the molecular mechanisms that underlie estrogen's effect on two specific forms of synaptic plasticity commonly studied in neurophysiology and the behavioral neurosciences: long-term potentiation and long-term depression. PMID- 11726237 TI - Is there savings for pavlovian fear conditioning after neurotoxic basolateral amygdala lesions in rats? AB - Considerable evidence indicates an important role for amygdaloid nuclei in both the acquisition and expression of Pavlovian fear conditioning. Recent reports from my laboratory have focused on the impact of neurotoxic lesions of the basolateral complex of the amygdala (BLA) on conditional freezing behavior in rats. In these studies, I have observed severe effects of posttraining BLA lesions on the expression of conditional freezing even after extensive presurgical overtraining (25-75 trials). Moreover, I have found no evidence for sparing of fear memory (i.e., savings) in these rats when I assess their rate of reacquisition relative to BLA rats receiving minimal training (1 trial). In these experiments, freezing behavior was assessed using a conventional time-sampling procedure and expressed as a response probability. Although this measure is well established in the literature, it is conceivable that it is not sensitive to spared memory in rats with BLA lesions. To address this issue, I present a more detailed analysis of freezing behavior that quantifies latency to freeze, the number of freezing bouts, the duration of freezing bouts, and the probability distribution of bout lengths. I also include control data from untrained (no shock) rats. Consistent with my earlier reports, I find no evidence of savings of fear memory in rats with neurotoxic BLA lesions using several measures of freezing behavior. These results reiterate the conclusion that fear memory, as it is expressed in freezing behavior, requires neurons in the BLA. PMID- 11726238 TI - Remembrance of arguments past: how well is the glutamate receptor hypothesis of LTP holding up after 20 years? PMID- 11726239 TI - Hippocampal theta oscillations and classical conditioning. AB - Studies are reviewed that support a hypothesized role for hippocampal theta oscillations in the neural plasticity underlying behavioral learning. Begun in Richard F. Thompson's laboratory in the 1970s, these experiments have documented a relationship between free-running 3- to 7-Hz hippocampal slow waves (theta) and rates of acquisition in rabbit classical nictitating membrane (NM) conditioning. Lesion and drug manipulations of septohippocampal projections have affected NM and jaw movement conditioning in ways consistent with a theta-related brain state being an important modulator of behavioral acquisition. These findings provide essential empirical support for the recently developed neurobiological and computational models that posit an important role for rhythmic oscillations (such as theta) in cellular plasticity and behavioral learning. PMID- 11726240 TI - Cerebellar substrates for error correction in motor conditioning. AB - The authors evaluate a mapping of Rescorla and Wagner's (1972) behavioral model of classical conditioning onto the cerebellar substrates for motor reflex learning and illustrate how the limitations of the Rescorla-Wagner model are just as useful as its successes for guiding the development of new psychobiological theories of learning. They postulate that the inhibitory pathway that returns conditioned response information from the cerebellar interpositus nucleus back to the inferior olive is the neural basis for the error correction learning proposed by Rescorla and Wagner (Gluck, Myers, & Thompson, 1994; Thompson, 1986). The authors' cerebellar model expects that behavioral processes described by the Rescorla-Wagner model will be localized within the cerebellum and related brain stem structures, whereas behavioral processes beyond the scope of the Rescorla Wagner model will depend on extracerebellar structures such as the hippocampus and related cortical regions. Simulations presented here support both implications. Several novel implications of the authors' cerebellar error correcting model are described including a recent empirical study by Kim, Krupa, and Thompson (1998), who verified that suppressing the putative error correction pathway should interfere with the Kamin (1969) blocking effect, a behavioral manifestation of error correction learning. The authors also discuss the model's implications for understanding the limits of cerebellar contributions to associative learning and how this informs our understanding of hippocampal function in conditioning. This leads to a more integrative view of the neural substrates of conditioning in which the authors' real-time circuit-level model of the cerebellum can be viewed as a generalization of the long-term memory module of Gluck and Myers' (1993) trial-level theory of cerebellar-hippocampal interaction in motor conditioning. PMID- 11726241 TI - MRI-assessed volume of cerebellum correlates with associative learning. AB - Richard F. Thompson's cerebellar model of classical eyeblink conditioning highlights Purkinje cells in cerebellar cortex and principal cells in the deep cerebellar nucleus as the integrating cells for acquisition of conditioned responses (CRs). CR acquisition is significantly slower in rabbits with lesions to cerebellar cortex and in Purkinje cell-deficient mice that lose all cerebellar cortical Purkinje cells. Purkinje cells are the largest neurons in the cerebellum and contribute significantly to cerebellar volume. Magnetic resonance imaging (MRI) was used to assess cerebellar volume in humans. Cerebellar volume was related to eyeblink conditioning (400-ms delay procedure) in 8 adults (21-35 years) and compared to 8 older adults (77-95 years) tested previously (Woodruff Pak, Goldenberg, Downey-Lamb, Boyko, & Lemieux, 2000). In the young adult sample, there was a high correlation between percentage of CRs in a session and cerebellar volume (corrected for total intracranial volume [TIV], r =.58, p =.066). There were statistically significant age differences in cerebellar volume, t(14) = 8.96, p <.001, and percentage of CRs, t(14) = 3.85, p <.002, but no age difference in TIV. Combining the young and older adult sample, the correlation between percentage of CRs and cerebellar volume (corrected for TIV) was.832 (p <.001). Cerebellar volume showed age-related deficits likely due to Purkinje cell loss. Individual differences in classical eyeblink conditioning are associated with differences in cerebellar volume, supporting Thompson's model of a cerebellar cortical role in facilitating this form of associative learning. PMID- 11726242 TI - Projection of the magnocellular red nucleus to the region of the accessory abducens nucleus in the rabbit. AB - The projection of the magnocellular red nucleus (RNm) to the region of the accessory abducens nucleus (AABD) was traced in rabbit using the bidirectional tracer wheat germ agglutinin-horseradish peroxidase (WGA-HRP). In one set of animals, recordings of antidromic responses from RNm neurons elicited by electrical stimulation of the rubrospinal tract were used to localize injections of WGA-HRP for orthograde labeling of RNm terminals. In a different set of animals, horseradish peroxidase was injected into the retractor bulbi muscle to retrogradely label motoneurons of the AABD. The positions of RNm fibers and terminals were examined and compared to the locations and distribution of AABD cell bodies and labeled dendrites. Analyses revealed that along the entire rostrocaudal extent of the AABD, RNm efferents terminate primarily lateral to, or in the lateral aspects of, labeled motoneurons. For the rostral AABD, RNm efferents terminate only lateral to the nucleus. Although the terminals are not positioned to contact cell bodies of the AABD, they could overlap with dendrites that extend in the lateral direction. RNm efferents terminate more extensively within the posterior AABD, overlapping within both dendritic and cell body regions of the nucleus. Even in this posterior region, however, RNm efferents were distributed primarily over the lateral half of the nucleus. These data show that RNm can monosynaptically influence the AABD, through primarily its lateral and posterior aspects. Our findings also show that a major target of RNm efferents is the reticular cell population located lateral to the AABD, suggesting that the RNm also may affect AABD motoneuronal output indirectly through its projection to reticular cells. PMID- 11726243 TI - Differential effect of TEA on long-term synaptic modification in hippocampal CA1 and dentate gyrus in vitro. AB - The effectiveness of tetraethylammonium (TEA) and high-frequency stimulation (HFS) in inducing long-term synaptic modification is compared in CA1 and dentate gyrus (DG) in vitro. High-frequency stimulation induces long-term potentiation (LTP) at synapses of both perforant path-DG granule cell and Schaffer collateral CA1 pyramidal cell pathways. By contrast, TEA (25 mM) induces long-term depression in DG while inducing LTP in CA1. The mechanisms underlying the differential effect of TEA in CA1 and DG were investigated. It was observed that T-type voltage-dependent calcium channel (VDCC) blocker, Ni2+ (50 microM), partially blocked TEA-induced LTP in CA1. A complete blockade of the TEA-induced LTP occurred when Ni2+ was applied together with the NMDA receptor antagonist, D APV. The L-type VDCC blocker, nifidipine (20 microM), had no effect on CA1 TEA induced LTP. In DG of the same slice, TEA actually induced long-term depression (LTD) instead of LTP, an effect that was blocked by D-APV. Neither T-type nor L type VDCC blockade could prevent this LTD. When the calcium concentration in the perfusion medium was increased, TEA induced a weak LTP in DG that was blocked by Ni2+. During exposure to TEA, the magnitude of field EPSPs was increased in both CA1 and DG, but the increase was substantially greater in CA1. Tetraethylammonium application also was associated with a large, late EPSP component in CA1 that persisted even after severing the connections between CA3 and CA1. All of the TEA effects in CA1, however, were dramatically reduced by Ni2+. The results of this study indicate that TEA indirectly acts via both T-type VDCCs and NMDA receptors in CA1 and, as a consequence, induces LTP. By contrast, TEA indirectly acts via only NMDA receptors in DG and results in LTD. The results raise the possibility of a major synaptic difference in the density and/or distribution of T-type VDCCs and NMDA receptors in CA1 and DG of the rat hippocampus. PMID- 11726244 TI - Regulation and mechanism of L-type calcium channel activation via V1a vasopressin receptor activation in cultured cortical neurons. AB - We have sought to elucidate the biochemical mechanisms that underlie the memory enhancing properties of the neural peptide vasopressin. Toward that goal we have investigated vasopressin induction of calcium signaling cascades, long held to be involved in long-term memory function, in neurons derived from the cerebral cortex, a brain region associated with long-term memory. Our previous studies demonstrated that in cultured cortical neurons, V1a vasopressin receptor (V1aR) activation resulted in a sustained rise in intracellular calcium concentration that was dependent on calcium influx (Son & Brinton, 1998). To investigate the mechanism of V1aR-induced calcium influx, we investigated V1aR activation of the calcium channel subtype(s) in cortical neurons cultured from Sprague-Dawley rat embryonic day 18 fetuses. The results of these analyses demonstrated that the L type calcium channel blocker nifedipine blocked 250 nM V1 vasopressin receptor agonist (V1 agonist)-induced calcium influx. Intracellular calcium imaging analyses using fura-2AM demonstrated that blockade of L-type calcium channels prevented the 250 nM V1 agonist-induced rise in intracellular calcium concentration. These results indicate that the influx of extracellular calcium via L-type calcium channels is an essential step in the initiation of the V1 agonist-induced rise in intracellular calcium concentration. To determine the mechanism of V1aR activation of L-type calcium channels, regulatory components of the phosphatidylinositol signaling pathway were investigated. The results of these analyses demonstrated that V1 agonist-induced calcium influx was blocked by both a phospholipase C inhibitor (U-73122) and a protein kinase C inhibitor (bisindolylmaleimide I). Further analysis of V1aR activation of protein kinase C (PKC) demonstrated that V1 agonist induced PKC activity within 1 min of exposure in cultured cortical neurons. These data indicate that in cultured cortical neurons, V1aR activation regulates the influx of extracellular calcium via L-type calcium channel activation through a protein kinase-C-dependent mechanism. The results of these studies provide biochemical mechanisms by which vasopressin could enhance memory function. Those mechanisms include a complex cascade that is initiated by activation of the phosphatidylinositol pathway, activation of protein kinase C, followed by phosphorylation of L-type calcium channels to initiate the influx of extracellular calcium to activate a cascade of calcium dependent release of intracellular calcium. PMID- 11726245 TI - Lesions in the central nucleus of the amygdala: discriminative avoidance learning, discriminative approach learning, and cingulothalamic training-induced neuronal activity. AB - The amygdala is critically involved in discriminative avoidance learning. Large lesions of the amygdala block discriminative avoidance learning and abolish cingulothalamic training-induced neuronal activity. These results indicated that amygdalar processing is critical for cingulothalamic plasticity. The larger lesions did not allow differentiation of the specific functioning of various amygdalar nuclei. Anatomical analysis showed that damage in the central (CE) nucleus of the amygdala was correlated with the severity of the behavioral deficit. The present study was carried out to determine whether smaller lesions, centered in the CE nucleus, would impair discriminative avoidance learning and block cingulothalamic plasticity. In addition, the possible role of the CE nucleus in appetitively motivated discriminative approach learning was examined for the first time. New Zealand White rabbits with CE nuclear lesions were first trained in the discriminative approach task. After attaining asymptotic performance, discriminative avoidance training sessions were alternated with continuing approach training sessions, one session each day. The rabbits with lesions were severely impaired in avoidance learning but showed no impairment of approach learning. Surprisingly, the attenuating effects of the lesions on cingulothalamic training-induced neuronal activity were more prevalent during approach learning than during avoidance learning. These results indicated that avoidance learning can be impaired by lesions centered in the CE nucleus that leave cingulothalamic plasticity largely intact and that the CE nucleus is involved in extra-cingulothalamic learning processes. PMID- 11726246 TI - Effects of early hippocampal lesions on trace, delay, and long-delay eyeblink conditioning in developing rats. AB - The effects of bilateral hippocampal aspiration lesions on later acquisition of eyeblink conditioning were examined in developing Long-Evans rat pups. Lesions on postnatal day (PND) 10 were followed by evaluation of trace eyeblink conditioning (Experiment 1) and delay eyeblink conditioning (Experiment 2) on PND 25. Pairings of a tone conditioned stimulus (CS) and periocular shock unconditioned stimulus (US, 100 ms) were presented in one of three conditioning paradigms: trace (380 ms CS, 500 ms trace interval, 880 ms interstimulus interval [ISI]), standard delay (380 ms CS, 280 ms ISI), or long delay (980 ms CS, 880 ms ISI). The results of two experiments indicated that hippocampal lesions impaired trace eyeblink conditioning more than either type of delay conditioning. In light of our previous work on the ontogeny of trace, delay, and long-delay eyeblink conditioning (Ivkovich, Paczkowski, & Stanton, 2000) showing that trace and long delay eyeblink conditioning had similar ontogenetic profiles, the current data suggest that during ontogeny hippocampal maturation may be more important for the short-term memory component than for the long-ISI component of trace eyeblink conditioning. The late development of conditioning over long ISIs may depend on a separate process such as protracted development of cerebellar cortex. PMID- 11726247 TI - The role of the hippocampus in trace conditioning: temporal discontinuity or task difficulty? AB - It is well established that the hippocampal formation is critically involved in the acquisition of trace memories, a paradigm in which the conditioned (CS) and unconditioned stimuli (US) are separated by a temporal gap (Solomon et al., 1986). The structure is reportedly not critical for the acquisition of delay memories, where the CS and the US overlap in time (Berger & Orr, 1983; Schmaltz & Theios, 1972). Based on these results, it is often stated that the hippocampus is involved in "filling the gap" or otherwise associating the two stimuli in time. However, in addition to the presence of a temporal gap, there are other differences between trace and delay conditioning. The most apparent difference is that animals require many more trials to learn the trace task, and thus it is inherently more difficult than the delay task. Here, we tested whether the hippocampus was critically involved in delay conditioning, if it was rendered more difficult such that the rate of acquisition was shifted to be analogous to trace conditioning. Groups of rats received excitotoxic lesions to the hippocampus, sham lesions or were left intact. Using the same interstimulus intervals (ISI), control animals required more trials to acquire the trace than the delay task. As predicted, animals with hippocampal lesions were impaired during trace conditioning but not delay conditioning. However, when the delay task was rendered more difficult by extending the ISI (a long delay task), animals with hippocampal lesions were impaired. In addition, once the lesioned animal learned the association between the CS and the US during delay conditioning, it could learn and perform the trace CR. Thus, the role of the hippocampus in classical conditioning is not limited to learning about discontiguous events in time and space; rather the structure can become engaged simply as a function of task difficulty. PMID- 11726248 TI - Dr. Patrick O'Shea: a man for all seasons. PMID- 11726249 TI - The relationship between field tests of anaerobic power and 10-km run performance. AB - The purpose of this study was to investigate the relationship between several field tests of anaerobic power and distance running performance. Thirty-six trained runners (20 men and 16 women; mean +/- SD age, 27.9 +/- 5.7 years) participated in this study. Tests of anaerobic power consisted of a 50-m sprint, vertical jumps from a static take-off position and with a countermovement, a plyometric leap test, and a 300-m sprint. The results indicated that gender, height, weight, percent body fat, 50-m sprint time, the height and power of both types of vertical jumps, plyometric leap distance, and the 300-m sprint time were significantly correlated with 10-km run time (p < or = 0.05) in the total subject pool (N = 36). Stepwise multiple regression identified the plyometric leap distance to explain 73.9% of the variance in run time. When combined with 300-m sprint time, 77.9% of the variance (standard error of the estimate, 2.92 minutes) was explained. The regression equation developed is Y' (10-km time) = 57.22 - 5.15(plyometric leap distance in meters) + 0.27(300-m sprint time in seconds). The results indicate that anaerobic power is significantly related to distance running performance and may explain a meaningful percentage of variability in 10 km run time. Therefore, it may be beneficial for distance runners to supplement aerobic training with some power and speed development such as plyometrics and sprinting. PMID- 11726250 TI - Creatine loading, resistance exercise performance, and muscle mechanics. AB - We tested the null hypothesis that creatine monohydrate loading (20 g per day for 7 days, n = 18) would not alter resistance exercise performance, isometric strength, or in vivo contractile properties of the quadriceps femoris muscle compared with loading with placebo (n = 13) in resistance-trained subjects. For the entire study group, the 1 repetition maximum (1RM) and 5-set performance (the number of repetitions) for unilateral, dynamic knee extension increased slightly (2% and 5%, respectively) after dietary supplementation, and these responses did not differ by condition. Maximal voluntary isometric torque and the rate of torque development did not change. During electromyostimulation, torque development and relaxation time were also unaffected. Our data suggest that creatine loading does not augment unilateral strength or multiset resistance exercise performance for knee extensions compared with placebo loading. PMID- 11726251 TI - Effect of maximal aerobic power on match performance in elite soccer referees. AB - The aim of this study was to verify whether referees' maximal aerobic power could influence competitive match exercise intensity. Eight elite-level referees (mean +/- SD age, 37.6 +/- 3.4 years) were each observed during 2 Serie A matches (n = 16), and the mean of each match activity was used for analysis. Match activities were monitored with encoder-equipped cameras. .V(O)(2)max was assessed under field conditions with a portable device during a progressive multistage protocol. The .V(O)(2)max of each subject was expressed in absolute terms (.V(O)(2)max(ab), L.min(-1)), relative terms (.V(O)(2)max (rel), ml.kg(-1).min(-1)), and independent from body mass (.V(O)(2)max (ind), ml.kg(-0.75).min(-1)). Significance was set at p < or = 0.05 for all measurements. Total distance covered during the match correlated significantly with .V(O)(2)max (ind) (r = 0.77) and .V(O)(2)max (ab) (r = 0.87). .V(O)(2)max (ind), .V(O)(2)max (rel), and .V(O)(2)max (ab) correlated negatively (p < 0.05) with the time spent standing still during the match (r = -0.90, -0.85, and -0.86, respectively, p < 0.05). During the first half of the match, referees with higher levels of .V(O)(2)max (rel) (r = 0.93, p < 0.05) and .V(O)(2)max (ind) (r = 0.90, p < 0.05) were able to cover more distance at medium intensity. In the second half of the match, medium-intensity distance and low-intensity time correlated significantly (p < 0.05) with .V(O)(2)max (ab) (r = 0.76) and .V(O)(2)max (rel) (r = 0.75), respectively. The results demonstrate the positive influence of .V(O)(2)max in determining both the total amount of distance covered and the exercise intensity during a game. Higher levels of .V(O)(2)max may provide referees the opportunity to be more active and therefore closer to the action on the field during the game. PMID- 11726252 TI - Effect of heavy dynamic resistive exercise on acute upper-body power. AB - There is limited research to support the notion that heavy resistive exercise immediately before a power movement may acutely enhance performance. Upper-body acute power enhancement during a stretch-shorten cycle (SSC) movement has not been previously investigated. The aim of this project was to determine whether a set of 5 repetitions of 5 repetition maximum (RM) bench press preceding explosive push-ups would significantly influence indicators of power: impulse and maximum rate of force development. Subjects randomly performed either explosive push-ups only over a force platform or a set of 5RM bench press before the explosive push ups. There were no significant differences for any of the force platform data when the explosive push-ups were preceded by a set of 5RM bench press. It appears that heavy dynamic resistive upper-body exercise on its own before an upper-body power activity is inadequate in augmenting short-term power. PMID- 11726253 TI - Validity and reliability of a medicine ball explosive power test. AB - The purpose of this study was to evaluate the validity and reliability of a medicine ball throw test to assess explosive power. Twenty competitive sand volleyball players (10 male players, 10 female players) performed a medicine ball throw and a standard countermovement vertical jump. The subjects attended 2 sessions; at each session, 3 attempts of each test were completed. The movement pattern for the medicine ball throw was a backward overhead toss. To standardize for body weight, a power index was calculated for the countermovement vertical jump using the Lewis formula. Validity was assessed using the best score for both the throw and the jump, and reliability was assessed using the best score from each session. There was a strong correlation between the distance of the medicine ball throw and the power index for the countermovement vertical jump (r = 0.906, p < 0.01). For the countermovement vertical jump, the test-retest reliability was 0.993 (p < 0.01), and for the medicine ball throw, the test-retest reliability was 0.996 (p < 0.01). These findings suggest that the medicine ball throw test is a valid and reliable test for assessing explosive power for an analogous total body movement pattern and general athletic ability. PMID- 11726254 TI - The effect of concentric isokinetic strength training of the quadriceps femoris on electromyography and muscle strength in the trained and untrained limb. AB - The purpose of the present investigation was to examine the effects of unilateral concentric isokinetic leg extension training on peak torque (PT) and electromyographic (EMG) responses in the trained and untrained limbs. Twenty adult men were randomly assigned to a training (TRN, n = 11) or control (CTL, n = 9) group. The TRN group performed 6 sets of 10 leg extensions 3 days per week for 12 weeks at a velocity of 90 degrees.s(-1). All subjects were tested every 4 weeks for PT and EMG responses of both legs at a velocity of 90 degrees.s(-1). The 3-way mixed factorial analysis of variance (ANOVA) indicated a significant (p < 0.05) increase in PT over the 12 weeks in both the trained and untrained limb for the TRN group but no significant change in PT in either limb for the CTL group. The results of the 3-way ANOVA for the EMG data indicated no significant change in EMG amplitude in the trained or untrained limb for the TRN or CTL group. The increase in PT in the absence of a change in EMG may result from hypertrophic factors and/or changes in the other muscles or muscle groups involved in leg extension. PMID- 11726255 TI - The effect of muscle hypoperfusion-hyperemia on repetitive vertical jump performance. AB - Alterations in skeletal muscle blood flow can greatly influence exercise performance. Brief periods of arterial hypoperfusion and subsequent hyperemia (hypoperfusion-hyperemia) have been shown to decrease the rate of skeletal muscle fatigue in a model of repeated, isometric wrist flexion exercise. However, the mechanism by which hypoperfusion-hyperemia influences dynamic motor skills remains unknown. The purpose of this study was to determine the effects of brief hypoperfusion-hyperemia (by femoral cuff occlusion) on repetitive vertical jump performance. Recreationally trained men and women (n = 10, mean +/- SD age, 25 +/ 2 years), performed 2 randomly assigned jumping trials, each consisting of 40 maximal effort vertical jumps. Jump height was videotaped on a Sony digital video recorder and analyzed with the Scion Images program. Trial 1 consisted of 40 vertical jumps without femoral artery occlusion. Trial 2 consisted of 40 vertical jumps preceded by femoral artery cuff occlusion for 90 seconds at 200 mm Hg, followed by 10 seconds of hyperemia before jumping. For both trials, the rate of decline in power output in men and women was approximately 20%. Hypoperfusion hyperemia had no significant effect on vertical jumping power output, perhaps because additional muscle groups used to jump vertically (e.g., gluteals and arms) were not occluded. These results warrant further research on the effect of hypoperfusion-hyperemia on strength and power measures. PMID- 11726256 TI - Physical fitness qualities of professional rugby league football players: determination of positional differences. AB - A total of 146 professional rugby league football players, contracted to 2 teams competing in England (n = 45) and Australia (n = 101), participated in this study. All players completed the following series of physical fitness performance tests: 1 repetition maximum squat and bench press, 15- and 40-m sprint, agility run, 5-minute run for distance, 60-second sit-up, 30-second plyometric push-up, and measurement of body weight and subcutaneous skinfold (4 sites). Analysis of variance with a criterion alpha level of p < 0.05, was used to determine if any significant difference could be found when grouping players into 3 different positional categories typically identified in the sport. There were a number of significant differences with respect to test results between categories, and this was apparent for all 3 systems of categorization. On the basis of these findings, we recommend that to more efficiently structure the physical fitness training of players, the players should be grouped either according to the 2 broad positional categories of forwards or backs or according to the 4 categories of forwards, distributors, adjustables, and outside backs. Grouping players according to the 9 specific positions played on the team is not warranted. PMID- 11726257 TI - Effects of different resistance training protocols on upper-body strength and endurance development in children. AB - This study examined the effects of 4 different resistance training protocols on upper-body strength and local muscle endurance development in children. Untrained boys and girls (mean +/- SD age, 8.1 +/- 1.6 years) trained twice per week for 8 weeks using child-sized weight machines and medicine balls weighing 1-2.5 kg. In addition to general conditioning exercises, subjects in each exercise group performed 1 set of the following exercise protocols for upper-body conditioning: 6-8 repetitions with a heavy load on the chest press exercise (HL, n = 15); 13-15 repetitions with a moderate load on the chest press exercise (ML, n = 16); 6-8 repetitions with a heavy load on the chest press exercise immediately followed by 6-8 medicine ball chest passes (CX, n = 12); or 13-15 medicine ball chest passes (MB, n = 11). Twelve children served as nontraining controls (CT). After training, only the ML and CX groups demonstrated significant (p < 0.05) improvements in 1RM chest press strength (16.8% and 16.3%, respectively) as compared with the CT group. Local muscle endurance, as determined by the number of repetitions performed posttraining on the chest press exercise with the pretraining 1RM load, significantly increased in the ML group (5.9 +/- 3.2 repetitions) and CX group (5.2 +/- 3.6 repetitions) as compared with the CT group. In terms of enhancing the upper-body strength and local muscle endurance of untrained children, these findings favor the prescription of higher-repetition training protocols during the initial adaptation period. PMID- 11726258 TI - Implementation of open and closed kinetic chain quadriceps strengthening exercises after anterior cruciate ligament reconstruction. AB - When working with athletes that have recently undergone anterior cruciate ligament (ACL) reconstructive surgery, a common goal of athletic trainers, strength and conditioning coaches, physicians, and physical therapists is to restore quadriceps strength while protecting the reconstructed ACL and patellofemoral joint from unnecessary stresses. Quadriceps strengthening exercises are often referred to as occurring in the open kinetic chain (OKC) or closed kinetic chain (CKC). Currently, there is little agreement in the literature as to whether only CKC exercises or a combination of OKC and CKC exercise should be performed after ACL reconstruction to strengthen the quadriceps. We believe that a combination of OKC and CKC exercises can be used to effectively and safely strengthen the quadriceps after ACL reconstruction. The purposes of this review are to examine the scientific literature currently available for the effects of OKC and CKC exercise on ACL strain and patellofemoral joint stress, and to present a sound rationale for using a combination of OKC and CKC exercises for quadriceps strengthening after ACL reconstruction. On the basis of our review, both OKC and CKC exercises can be modified and implemented for quadriceps strengthening after ACL reconstruction without causing excessive ACL strain or patellofemoral joint stress. PMID- 11726259 TI - Preferred exertion across three common modes of exercise training. AB - The use of self-selected intensities of exercise may increase adherence to exercise programs by allowing the participant more freedom to choose activities that are enjoyable. However, self-selected intensities may vary across exercise modes, and participants may not choose an intensity that is adequate to produce health benefits. The purpose of this study was to determine influence of exercise mode on self-selected exercise intensities. Eighteen subjects (12 men and 6 women) between the ages of 18 and 25 participated in this study. Preferred intensity tests were performed for 3 modes of exercise (treadmill, cycle ergometer, and stairstepper). .V(O)(2) values were obtained continuously and 1 minute averages were recorded at minutes 5, 10, 15, and 20 for each submaximal test. Comparisons were made using a repeated-measures analysis of variance (mode, time, mode x time). Scheffe's F test was used to test simple effects. There was a significant increase in the relative .V(O)(2) for all 3 modes of exercise across the 20-minute trials (p = 0.0001). Relative .V(O)(2) (%.V(O)(2)peak) increased from 52.20 to 64.71% for cycle exercise, 43.27 to 63.25% for the treadmill, and 47.16 to 61.17% for stairstepping. The average relative .V(O)(2) for the cycle ergometer (60.40 +/- 15.55%) was significantly higher (p = 0.02) than both the treadmill (53.65 +/- 18.36%) and stairstepper (54.66 +/- 11.98%). Relative heart rate (HR) (%HRR) for the stairstepper (80.21 +/- 9.67%) and the cycle ergometer (80.03 +/- 10.59%) were significantly higher than the treadmill (74.77 +/- 13.13%) (p = 0.0003). There were no significant differences in rating of perceived exertion (RPE) among the 3 modes of exercise. Similar RPE values were reported for the stairstepper (12.79 +/- 2.97), cycle ergometer (12.57 +/- 2.90), and the treadmill (12.50 +/- 2.87). The results indicate that subjects allowed to choose exercise intensity by self-selection chose work rates that were within the moderate range of American College of Sports Medicine guidelines of 50-85% .V(O)(2)max for treadmill, cycle ergometer, and stairstepping exercise. PMID- 11726260 TI - Relative activity of abdominal muscles during commonly prescribed strengthening exercises. AB - The purpose of this study was to determine the relative electromyographic (EMG) activity of the upper and lower rectus abdominis and the external oblique muscles during 5 commonly performed abdominal strengthening exercises. Twenty-five healthy subjects participated in the study. EMG data were collected under isometric and dynamic conditions. The reverse curl resulted in the greatest amount of lower rectus activity, the v-sit and reverse curl exercises resulted in the greatest amount of external oblique activity, and the trunk curl, reverse curl, trunk curl with a twist, and v-sit all resulted in similar amounts of upper rectus EMG activity. The vacuum exercise resulted in moderate levels of external oblique EMG activity but very low levels of activity in the rectus abdominis. Our findings support the concept that abdominal strengthening exercises can differentially activate various abdominal muscle groups, but contradict some traditionally held assumptions regarding the effects of specific exercises. PMID- 11726261 TI - Safety measures of L-carnitine L-tartrate supplementation in healthy men. AB - The purpose of this investigation was to examine the effects of ingestion of L CARNIPURE (L-carnitine L-tartrate [LCLT]) on alterations in a complete blood cell profile and in circulating metabolic enzymes. Using a balanced, placebo (P), cross-over design (1 week washout), 10 healthy, active men volunteered and acted as their own control taking either a P or LCLT supplement (3 g.day(-1)) for 3 weeks. Postabsorptive morning blood samples were obtained both before and after 21 days of P and LCLT supplementation. Serum samples were analyzed for clinical chemistries including a complete chemistry panel with markers of liver and renal function along with various minerals and electrolytes. In addition, whole blood was analyzed for a complete blood count with differential. It was determined that there were no statistically significant differences between the LCLT and the placebo conditions for any of the variables examined. The results of this study suggest that LCLT, when used as a dietary supplement, has no adverse effects on metabolic and hematological safety variables in normally healthy men. PMID- 11726262 TI - Effect of beta-hydroxy beta-methylbutyrate on the onset of blood lactate accumulation and V(O)(2) peak in endurance-trained cyclists. AB - The purpose of this study was to determine the effect of beta-hydroxy beta methylbutyrate (HMB) supplementation on maximal oxygen consumption (.V(O)(2)peak) and the onset of blood lactate accumulation (OBLA) in endurance-trained cyclists. Eight cyclists randomly (double blind) completed 3 2-week supplementation periods (HMB, 3g.day(-1); leucine [LEU], 3g.day(-1); placebo [CON], 3g.day(-1)) followed by a 2-week washout period. Testing consisted of a graded cycle ergometry test to measure .V(O)(2)peak and OBLA, the .V(O)(2) at 2 mM blood lactate. .V(O)(2)peak was unaffected by HMB (4.0 +/- 1.4%), LEU (-1.9 +/- 1.3%), and CON (-2.6 +/- 2.6%). HMB resulted in a greater time to reach .V(O)(2)peak, whereas LEU and CON did not affect this time (HMB, 3.6 +/- 1.5 min, LEU, -1.2 +/- 1.5 min; CON, -3.6 +/- 3.5 min). Lactate accumulation peak was unaffected by supplementation (HMB, 8.1 +/- 1.1 mM; LEU, 6.2 +/- 0.8 mM; CON, 7.5 +/- 1.3 mM). OBLA increased with HMB (9.1 +/- 2.4%) and LEU (2.1 +/- 1.5%), but not in the CON trial (0.75 +/- 2.1%). Blood glucose was significantly greater during the HMB trial compared with the LEU trial. It is concluded that HMB supplementation may have positive affects on performance by increasing the onset of blood lactate accumulation; however, the mechanism is unknown. PMID- 11726263 TI - Effects of short-term isokinetic training on standing long-jump performance in untrained men. AB - The purpose of this study was to examine the effects of a 6-week isokinetic training program on quadriceps and hamstrings peak torque (PT) and standing long jump (SLJ) performance. Twelve untrained men (age 31.4 +/- 4.2 years, mean +/- SD) were tested at a velocity of 1.75 rad.s(-1) (100 degrees.s(-1)), before and after a 6-week control period and on completion of a 6-week training program. Training consisted of 3 sets of 10 repetitions, 3 days per week, for 6 weeks. Repeated-measures analysis-of-variance analyses revealed that quadriceps PT increased, on average, by 10.5% as a result of training (p < 0.01). No significant changes in hamstrings PT (p = 0.062) and SLJ performance occurred as a result of training (p > 0.05). The major finding of this study was that PT gains subsequent to isokinetic resistance training did not influence functional performance. That open-chain training did not affect the performance of a closed chain activity is unsurprising, but on these grounds of nonfunctionality, the use of moderate velocity isokinetic dynamometry in rehabilitation and performance assessment for closed-chain sports needs addressing in future research. PMID- 11726264 TI - Tachycardia during resistance exercise: a case study. AB - A male weight-trained (WT) subject (A; age = 21 years, height = 193.6 cm, body weight = 113.4 kg, parallel barbell squat 10 repetition maximum [RM] = 183.7 kg) and a group of 18 similarly trained men (WT; X +/- SD; age = 24.7 +/- 3.0 years, height = 180.2 +/- 4.3 cm, body weight = 86.9 +/- 10.7 kg, 10 RM = 127.9 +/- 28.6 kg) were monitored during a barbell squat resistance exercise session (50% - 100% 10 RM) and a graded exercise test on a cycle ergometer. Compared with the WT group, heart rate (HR) for subject A was consistently > or =2 SD greater during resistance exercise (peak HR for 100% 10 RM = 212 b.min(-1) and 165.3 +/- 16.2 b.min(-1)). The graded exercise test resulted in similar HR responses for both A and controls. Subject A's augmented HR was present only during the pressure load of heavy resistance exercise, and not during the volume load of endurance exercise. The data suggest that subject A may be utilizing a different mechanism for heart rate regulation during resistance exercise. PMID- 11726265 TI - Dietary practices, attitudes, and physiological status of collegiate freshman football players. AB - The purpose of this study was to determine the dietary practices, attitudes, and physiological status of freshman collegiate football players. Thirty-one freshman football players at a National Collegiate Athletic Association division I school completed a self-administered nutrition screening questionnaire designed to determine their dietary practices and attitudes. Fasting blood samples were collected and height and weight were measured. The mean age of these athletes was 18 years. These players reported eating 3.6 times per day and on the average eating out 4.8 times per week. Fast food was the most common choice when eating out (55%). Of these athletes, 42% reported the use of dietary supplements, the most popular one being creatine (36%). Although more than 90% of the athletes recognized the importance of maintaining proper hydration status, greater than 50% believed that protein supplements were necessary for muscle growth and development, protein was the primary source of energy for muscle, and vitamin and mineral supplements increased energy levels. The plasma lipid profiles of the majority (76%) of the athletes were within normal ranges. The data suggest that these athletes may require education about healthy dietary practices and on the proper use of dietary supplements. This is not only important to help improve performance but to also promote healthy dietary practices in the long term. PMID- 11726266 TI - Effect of overhydration on time-trial swim performance. AB - The effect of hydration status on performance has not been adequately emphasized or examined in swimmers. Theoretically, moderate overhydration might reduce the proportionate fluid loss from the circulation during exercise of this nature. To explore this issue, 11 (5 women, 6 men) collegiate swimmers swam 2 183-m (200-yd) time trials (3 days apart) in alternate, randomized euhydrated (EUH) and overhydrated (OH) states. Pre-exercise plasma osmolality (EUH: 288.5 +/- 2.5 and OH: 284.6 +/- 3.3 mOsmol.kg(-1); p < 0.001), urine specific gravity (EUH: 1.022 +/- 0.003 and OH: 1.012 +/- 0.003; p < 0.001), and body weight (EUH: 72.1 +/- 9.3 and OH: 72.6 +/- 9.2 kg; p < 0.01) values distinguished the two hydration states of the swimmers. There was no difference (p > 0.05) between hydration states in postexercise plasma osmolality (EUH: 312.8 +/- 4.8 and OH: 307.2 +/- 9.9 mOsmol.kg(-1)), plasma volume (EUH: -16.5 +/- 10.0 and OH: -17.7 +/- 6.8 %Delta), plasma lactate (EUH: 18.6 +/- 3.6 and OH: 17.8 +/- 3.4 mmol.1(-1)), heart rate (EUH: 167 +/- 11 and OH: 166 +/- 16 beats.min(-1)), or perceived exertion (EUH: 16 +/- 1 and OH: 16 +/- 2) responses. Although performance time improved for 7 of the 11 swimmers during OH, there was not a statistically significant difference between the EUH (121.2 +/- 8.1 seconds) and OH (120.8 +/- 7.7 seconds) conditions. However, there was a modest bivariate correlation (r = -0.602; p < 0.05) between the change in body weight and change in performance time in going from the EUH to OH trials. These data demonstrated that overhydration provided no performance advantage for this group during a 183-m time-trial swim but emphasized the importance of adequate hydration in swim performance. PMID- 11726267 TI - Orientation and familiarization to 1RM strength testing in old and young women. AB - The purpose of this study was to compare the number of testing sessions required to achieve consistent 1 repetition maximum (1RM) strength measurements in untrained old and young women. Consistency of measurement was defined as consecutive 1 RM strength measures that increased by 1 kg or less. Untrained old (n = 6, age 66 +/- 5 years) and untrained young (n = 7, age 23 +/- 4 years) women were repeatedly strength-tested for bilateral concentric knee extension 1 RM strength until consecutive measurements were increased by no more than 1 kg. At least 48 hours of rest was allowed between 1 RM measurements. The old subjects required significantly more testing sessions (8-9 sessions) compared with the young subjects (3-4 sessions) to achieve the same absolute consistency of measurement (p < 0.05). Absolute increase in strength between the first and final testing sessions did not differ between groups (young = 11 +/- 4 kg and old = 13 +/- 2 kg) (p > 0.05). The relative increase was significantly greater in the older subjects (young = 12 +/- 5%; old = 22 +/- 4%) (p < 0.05). In conclusion, older subjects require more practice and familiarization and show greater relative increases in 1RM strength when compared with younger subjects of the same experience level. This is important to consider, especially when evaluating the magnitude of strength increase in response to resistance training. PMID- 11726268 TI - Is there a physiologic basis for creatine use in children and adolescents? AB - The purported ergogenic benefits of creatine for the adult population have been well documented. In able-bodied children and adolescents, there is a paucity of data on creatine use and the purported ergogenic effects of creatine. Only 1 study to date has investigated the ergogenic properties of creatine in the adolescent population. The purpose of this review was to try to establish a rationale for creatine use in the child and adolescent population. The limited literature available in this area did not provide a strong enough rationale from either a physiologic or performance perspective for creatine supplementation in these populations. However, significantly more research is required before definitive conclusions can be made. PMID- 11726269 TI - Clearance of extracellular and cell-associated amyloid beta peptide through viral expression of neprilysin in primary neurons. AB - Amyloid beta peptide (Abeta), the pathogenic agent of Alzheimer's disease (AD), is a physiological metabolite constantly anabolized and catabolized in the brain. We previously demonstrated that neprilysin is the major Abeta-degrading enzyme in vivo. To investigate whether or not manipulation of neprilysin activity in the brain would be an effective strategy for regulating Abeta levels, we expressed neprilysin in primary cortical neurons using a Sindbis viral vector and examined the effect on Abeta metabolism. The corresponding recombinant protein, expressed in the cell bodies and processes, exhibited thiorphan-sensitive endopeptidase activity, whereas a mutant neprilysin with an amino acid substitution in the active site did not show any such activity. Expression of the wild-type neprilysin, but not the mutant, led to significant decreases in both the Abeta40 and 42 levels in the culture media in a dose-dependent manner. Moreover, neprilysin expression also resulted in reducing cell-associated Abeta, which could be more neurotoxic than extracellular Abeta. These results indicate that the manipulation of neprilysin activity in neurons, the major source of Abeta in the brain, would be a relevant strategy for controlling the Abeta levels and thus the Abeta-associated pathology in brain tissues. PMID- 11726270 TI - Crystallization and preliminary X-ray analysis of a DNA primase from hyperthermophilic archaeon Pyrococcus horikoshii. AB - At the initiation of chromosomal DNA replication, DNA primases synthesize short RNA primers, which are subsequently elongated by DNA polymerases. To understand the structural basis for the primer synthesis by archaeal/eukaryotic-type primases, the gene of the DNA primase from hyperthermophilic archaeon Pyrococcus horikoshii was cloned and overexpressed in Escherichia coli as a fusion protein with a hexa-histidine tag at its amino terminus. The recombinant DNA primase was purified and crystallized by the hanging-drop vapor diffusion method at 293 K, with polyethylene glycol 8000 as the precipitant. The crystals belong to the P3(2)21 space group with unit-cell parameters a = b = 77.8, c = 129.6 A, and alpha = beta = 90 degrees, gamma = 120 degrees. Crystals of the selenomethionine derivative were obtained by means of a cross-seeding method using native crystals. The data for the native and selenomethionine-substituted crystals were collected to 1.8 and 2.2 A resolution, respectively, with synchrotron radiation at SPring-8 under flash-frozen conditions at 100 K. The four wavelength MAD data provided a phase to determine the structure of the primase at 2.2 A resolution. PMID- 11726271 TI - Molecular cloning and expression of Caenorhabditis elegans ERp57-homologue with transglutaminase activity. AB - Formation of cross-linking between proteins via a gamma-glutamyl-epsilon-lysine residue is an important process in many biological phenomena including apoptosis. Formation of this linkage is catalyzed by the enzyme transglutaminase, which is widely distributed from bacteria to the animal kingdom. The simple multi-cellular organism Caenorhabditis elegans also possesses transglutaminase activity associated with apoptosis [Madi, A. et al. (1998) Eur. J. Biochem. 253, 583-590], but no gene with significant homology to vertebrate or bacterial transglutaminases has been found in the C. elegans genome sequence database. On the other hand, protein disulfide isomerases were recently recognized as a new family of transglutaminases [Chandrashekar, R. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 531-536]. To identify the molecule with transglutaminase activity in C. elegans, we isolated from C. elegans a gene homologous to ERp57, which encodes a protein disulfide isomerase, expressed it in recombinant form, and characterized the transglutaminase and protein disulfide isomerase activities of the resultant protein. The C. elegans ERp57 protein had both enzyme activities, and the transglutaminase activity had similar characteristics to the activity in lysate of the whole worm. These results suggested that the ERp57 homologue was one of the substances with transglutaminase activity in C. elegans. PMID- 11726272 TI - Interference with interaction between eukaryotic translation initiation factor 4G and poly(A)-binding protein in Xenopus oocytes leads to inhibition of polyadenylated mRNA translation and oocyte maturation. AB - The interaction between eukaryotic translation initiation factor 4G (eIF4G) and the poly(A)-binding protein (PABP) facilitates translational initiation of polyadenylated mRNAs. It was shown recently that the expression of an eIF4GI mutant defective in PABP binding in Xenopus oocytes reduces polyadenylated mRNA translation and dramatically inhibits progesterone-induced oocyte maturation. These results strongly suggest that the eIF4G-PABP interaction plays a critical role in the translational control of maternal mRNAs during oocyte maturation. In the present work, we employed another strategy to interfere eIF4G-PABP interaction in Xenopus oocytes. The amino-terminal part of eIF4GI containing the PABP-binding site (4GNt-M1) was expressed in Xenopus oocytes. 4GNt-M1 could bind to PABP in oocytes, which suggests that 4GNt-M1 may evict PABP from the endogenous eIF4G. The expression of 4GNt-M1 resulted in reduction of polyadenylated mRNA translation. Furthermore, 4GNt-M1 inhibited progesterone induced oocyte maturation. In contrast, 4GNt-M2, in which the PABP-binding sequences were mutated to abolish the PABP-binding activity, could not inhibit polyadenylated mRNA translation or oocyte maturation. These results further support the idea that the eIF4G-PABP interaction is critical for translational regulation of maternal mRNAs in oocytes. PMID- 11726273 TI - Dissociation of Bax from a Bcl-2/Bax heterodimer triggered by phosphorylation of serine 70 of Bcl-2. AB - Serine 70 in the loop region of Bcl-2 is specifically phosphorylated by paclitaxel-treatment in tumor cells and BHK cells expressing Bcl-2. The phosphorylation of serine 70 of Bcl-2 (pS70-Bcl-2) peaks 24 to 48 h after paclitaxel treatment and accelerates apoptosis. Phosphorylation is effectively inhibited in the presence of actinomycin D or cycloheximide, which restore cell viability to the same level as control cells not expressing Bcl-2. These results indicate that paclitaxel-induced kinase(s) and/or its activator(s) are synthesized de novo and play an important role in paclitaxel-induced apoptosis by phosphorylating Bcl-2. In binding assays using the phosphorylation-specific antibody against pS70-Bcl-2, the induction of serine 70 phosphorylation 70 results in a loss of the binding ability of Bcl-2 to Bax, a pro-apoptotic partner, and induces subsequent cell death. When the pS70-Bcl-2 antibody was added to human breast cancer tissue, serine 70 phosphorylation was also detected, even prior to treatment with anticancer agents. Further study of breast cancers revealed 83% of tumors with high pS70-Bcl-2 expression responded to paclitaxel or docetaxel treatment, whereas 57% of those with low expression not respond. These findings suggest that pS70-Bcl-2 might be a predictive factor for prognosis and sensitivity to paclitaxel treatment for breast cancer. PMID- 11726274 TI - Side chain effect on ion channel characters of Aib rich peptides. AB - As models of ion channel proteins and naturally occurring pore-forming peptides, we designed a series of Aib rich peptides [Ac-(Aib-Xxx-Aib-Ala)(5)-NH(2) (Xxx = Lys, Glu, Ser, and Gly: BXBA-20)] to investigate the effects of the side chains of the amino acid residues Lys, Glu, Ser, and Gly on the conformation and electrophysiological properties of ion channels. The conformation of peptides and their affinity for phospholipid membranes were evaluated by CD spectroscopy. Patch-clamp experiments revealed that all BXBA-20 peptides form ion channels in DPhPC bilayers exhibiting clearly resolved transitions between the open and closed states. The channel forming frequency was in the order BKBA-20>BEBA 20>BSBA-20>BGBA-20. In the case of BKBA-20 and BEBA-20, the self-assembled conductive oligomers expressed homogeneous and voltage-independent single channel conductances. In contrast, heterogeneous conductance was observed in BSBA-20 and BGBA-20 ion channels under similar experimental conditions. From these results, we conclude that peptides with a high degree of helical conformation, high amphipathicity, high affinity for lipid membranes, and self-associating characters in vesicles are most suitable for inducing ion channels with a high frequency of occurrence. Moreover, BEBA-20, BSBA-20, and BGBA-20 channels were cation-selective, whereas the BKBA-20 channel was non-selective. PMID- 11726275 TI - Effects of calnexin deletion in Saccharomyces cerevisiae on the secretion of glycosylated lysozymes. AB - Disruption of the calnexin gene in Saccharomyces cerevisiae did not lead to gross effects on the levels of cell growth and secretion of wild-type hen egg white lysozymes (HEWL). To investigate the function of calnexin in relation to the secretion of glycoproteins, we expressed both stable and unstable mutant glycosylated lysozymes in calnexin-disrupted S. cerevisiae. The secreted amounts of stable mutant glycosylated lysozymes (G49N and S91T/G49N) were almost the same in both wild-type and calnexin-disrupted S. cerevisiae. In contrast, the secretion of unstable mutant glycosylated lysozymes (K13D/G49N, C76A/G49N, and D66H/G49N) greatly increased in calnexin-disrupted S. cerevisiae, although their secretion was very low in the wild-type strain. This indicates that calnexin may act in the quality control of glycoproteins. We further investigated the expression level of the mRNA of the molecular chaperones BiP and PDI, which play a major role in the protein folding process in the ER, when glycosylated lysozymes were expressed in wild-type and calnexin-disrupted S. cerevisiae. The mRNA concentrations of BiP and PDI were evidently increased when the glycosylated lysozymes were expressed in calnexin-disrupted S. cerevisiae. This observation indicates that BiP and PDI may be induced by the accumulation of unfolded glycosylated lysozymes due to the deletion of calnexin. PMID- 11726276 TI - Association of mouse sorting nexin 1 with early endosomes. AB - Sorting nexin 1 (SNX1) is a protein that binds to the cytoplasmic domain of plasma membrane receptors. We found that mouse sorting nexin 1 (SNX1) (521 amino acid residues) could partially rescue a yeast vam3 mutant defective in docking/fusion of vacuolar membranes. In mammalian cells, SNX1 is peripherally associated with membrane structures and localized immunochemically with EEA1, a marker protein of early endosomes. These results suggest that SNX1 regulates endocytic trafficking of plasma membrane proteins in early endosomes. Gel filtration of cell lysates and the purified recombinant protein, together with two-hybrid analysis, indicated that SNX1 self-assembles into a complex of approximately 300 kDa. PMID- 11726277 TI - Mixed lineage kinase LZK forms a functional signaling complex with JIP-1, a scaffold protein of the c-Jun NH(2)-terminal kinase pathway. AB - Leucine zipper-bearing kinase (LZK) is a novel member of the mixed lineage kinase (MLK) protein family, the cDNA of which was first cloned from a human brain cDNA library [Sakuma, H., Ikeda, A., Oka, S., Kozutsumi, Y., Zanetta, J.-P., and Kawasaki, T. (1997) J. Biol. Chem. 272, 28622-28629]. Several MLK family proteins have been proposed to function as MAP kinase kinase kinases in the c-Jun NH(2) terminal kinase (JNK)/stress-activated protein kinase (SAPK) pathway. In the present study, we demonstrated that, like other MLKs, LZK activated the JNK/SAPK pathway but not the ERK pathway. LZK directly phosphorylated and activated MKK7, one of the two MAPKKs in the JNK/SAPK pathway, to a comparable extent to a constitutive active form of MEKK1 (MEKK1DeltaN), suggesting a biological role of LZK as a MAPKKK in the JNK/SAPK pathway. Recent studies have revealed the essential roles of scaffold proteins in intracellular signaling pathways including MAP kinase pathways. JIP-1, one of the scaffold proteins, has been shown to be associated with MLKs, MKK7, and JNK [Whitmarsh, A.J., Cavanagh, J., Tournier, C., Yasuda, J., and Davis, R.J. (1998) Science 281, 1671-1674], suggesting the presence of a selective signaling pathway including LZK, MKK7, and JNK. Consistent with this hypothesis, we provided evidence that LZK is associated with the C-terminal region of JIP-1 through its kinase catalytic domain. In addition, LZK-induced JNK activation was markedly enhanced when LZK and JNK were co-expressed with JIP-1. These results constituted important clues for understanding the molecular mechanisms regulating the signaling specificities of various JNK activators under different cellular conditions. PMID- 11726278 TI - Effects of cobalt-substitution of the active zinc ion in thermolysin on its activity and active-site microenvironment. AB - Thermolysin is remarkably activated in the presence of high concentrations (1-5 M) of neutral salts [Inouye, K. (1992) J. Biochem. 112, 335-340]. The activity is enhanced 13-15 times with 4 M NaCl at pH 7.0 and 25 degrees C. Substitution of the active site zinc with other transition metals alters the activity of thermolysin [Holmquist, B. and Vallee, B.L. (1974) J. Biol. Chem. 249, 4601 4607]. Cobalt is the most effective among the transition metals and doubles the activity toward N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide. In this study, the effect of NaCl on the activity of cobalt-substituted thermolysin was examined. Cobalt-substituted thermolysin, with 2.8-fold increased activity compared with the native enzyme, is further activated by the addition of NaCl in an exponential fashion, and the activity is enhanced 13-15 times at 4 M NaCl. The effects of cobalt-substitution and the addition of salt are independent of each other. The activity of cobalt-substituted thermolysin, expressed as k(cat)/K(m), is pH-dependent and controlled by at least two ionizing residues with pK(a) values of 6.0 and 7.8, the acidic pK(a) being slightly higher compared to 5.6 of the native enzyme. These pK(a) values remain constant in the presence of 4 M NaCl, indicating that the electrostatic environment of cobalt-substituted thermolysin is more stable than that of the native enzyme, the acidic pK(a) of which shifts remarkably from 5.6 to 6.7 at 4 M NaCl. Zincov, a competitive inhibitor, binds more tightly to the cobalt-substituted than to native thermolysin at pH 4.9-9.0, probably because of its preference for cobalt in the fivefold coordination. The cobalt substitution has been shown to be a favorable tool with which to explore the active-site microenvironment of thermolysin. PMID- 11726279 TI - Adrenodoxin-cytochrome P450scc interaction as revealed by EPR spectroscopy: comparison with the putidaredoxin-cytochrome P450cam system. AB - The cholesterol side-chain cleavage reaction catalyzed by cytochrome P450scc comprises three consecutive monooxygenase reactions (22R-hydroxylation, 20S hydroxylation, and C(20)-C(22) bond scission) that produces pregnenolone. The electron equivalents necessary for the oxygen activation are supplied from a 2Fe 2S type ferredoxin, adrenodoxin. We found that 1:1 stoichiometric binding of oxidized adrenodoxin to oxidized cytochrome P450scc complexed with cholesterol or 25-hydroxycholesterol caused shifts of the high-spin EPR signals of the heme moiety at 5 K. Such shifts were not observed for the low-spin EPR signals. Ligation of CO or NO to the reduced heme of cytochrome P450scc complexed with reduced adrenodoxin and various steroid substrates did not cause any change in the axial EPR spectrum of the reduced iron-sulfur center at 77 K. These results are in remarkable contrast to those obtained for the cytochrome P450cam-d-camphor putidaredoxin ternary complex, suggesting that the mode of cross talk between adrenodoxin and cytochrome P450scc is very different from that in the Pseudomonas system. The difference may be primarily due to the location of the charged amino acid residues of the ferredoxins important for the interaction with the partner cytochrome P450. PMID- 11726280 TI - Apoptosis induction associated with cell cycle dysregulation by rice bran agglutinin. AB - Effects of rice bran agglutinin (RBA) on human monoblastic leukemia U937 cells were examined in comparison with those of wheat germ agglutinin (WGA) and Viscum album agglutinin (VAA). These lectins inhibit cell growth, and several lines of evidence indicate that the growth inhibition is caused by the induction of apoptosis. We observed that RBA induces chromatin condensation, externalization of membrane phosphatidylserine, and DNA ladder formation, features of apoptosis. DNA ladder formation was inhibited by a general inhibitor against caspases, which are known to play essential roles in apoptosis. Flow cytometric analysis revealed that RBA and WGA cause G2/M phase cell cycle arrest with increased expression of Waf1/p21, while cell cycle arrest was not observed for VAA. These data indicate that RBA induces apoptosis associated with cell cycle arrest in U937 cells, and suggest that the induction mechanism for RBA is similar to that for WGA, but different from that for VAA. PMID- 11726281 TI - Identification of active site residues in Bradyrhizobium japonicum acetyl-CoA synthetase. AB - Acetyl-CoA synthetase (ACS) catalyses the activation of acetate to acetyl-CoA in the presence of ATP and CoA. The gene encoding Bradyrhyzobium japonicum ACS has been cloned, sequenced, and expressed in Escherichia coli. The enzyme comprises 648 amino acid residues with a calculated molecular mass of 71,996 Da. The recombinant enzyme was also purified from the transformed E. coli. The enzyme was essentially indistinguishable from the ACS of B. japonicum bacteroids as to the criteria of polyacrylamide gel electrophoresis and biochemical properties. Based on the results of database analysis, Gly-263, Gly-266, Lys-269, and Glu-414 were selected for site-directed mutagenesis in order to identify amino acid residues essential for substrate binding and/or catalysis. Four different mutant enzymes (G263I, G266I, K269G, and E414Q) were prepared and then subjected to steady-state kinetic studies. The kinetic data obtained for the mutants suggest that Gly-266 and Lys-269 participate in the formation of acetyl-AMP, whereas Glu-414 may play a role in acetate binding. PMID- 11726282 TI - Effects of nucleotides on N-acetyl-d-glucosamine 2-epimerases (renin-binding proteins): comparative biochemical studies. AB - Renin-binding protein (RnBP) is an endogenous renin inhibitor originally isolated from porcine kidney as a complex of renin, so-called high molecular weight (HMW) renin. Our recent studies demonstrated that human RnBP is the enzyme N-acetyl-D glucosamine (GlcNAc) 2-epimerase [Takahashi, S. et al. (1999) J. Biochem. 125, 348-353]. We have purified recombinant human, rat, and porcine RnBPs expressed in Escherichia coli JM 109 cells. The purified recombinant RnBPs existed as dimers and inhibited porcine renin activity strongly. On the other hand, porcine renin inhibited recombinant GlcNAc 2-epimerase activities. The human GlcNAc 2-epimerase activity could not be detected in the absence of a nucleotide, whereas ATP, dATP, ddATP, ADP, and GTP enhanced the human GlcNAc 2-epimerase activity. Other nucleotides had no effect on human GlcNAc 2-epimerase activity. Rat and porcine GlcNAc 2-epimerases were activated by several nucleotides. Nucleotides that enhance the activity of GlcNAc 2-epimerases protect these enzymes against degradation by thermolysin. These results indicate that mammalian RnBPs have GlcNAc 2-epimerase activity and that nucleotides are essential for formation of the catalytic domain of the enzyme. PMID- 11726283 TI - Characterization of phagosomal subpopulations along endocytic routes in osteoclasts and macrophages. AB - Modifications occurring during the transformation of phagosomes into mature phagolysosomes were investigated in osteoclast-like cells (OCLs) and macrophages using latex beads as markers for the isolation of phagosomal compartments (LBC) at different time points after phagocytosis. In OCLs, newly formed LBC acquired cathepsin K, tartarate-resistant phosphatase (TRAP), lysosome-associated membrane protein-1 (Lamp-1), and cathepsin D, and rapidly lost annexin II in a time dependent manner. The levels of Rab7 and c-Src in OCLs initially increased and then gradually decreased during the transformation from early to late endosomal LBC or phagolysosomes. Receptor activator of NF-kappaB (RANKL) significantly increased the LBC levels of cathepsin K, TRAP, and c-Src, whereas calcitonin decreased the LBC levels of cathepsin K, TRAP, and Rab7, indicating that the transformation of early to late endosomal elements and lysosomes in OCLs is also regulated by osteoclastogenesis regulatory factors. On the other hand, changes in the LBC levels of Lamp-1, cathepsin D, and annexin II in macrophages were comparable to those in OCLs. However, contrary to osteoclastic LBC, Rab7 levels of macrophage LBC decreased in a time-dependent manner. Macrophage LBC were devoid of cathepsin K, TRAP, and c-Src in all transformation stages. These observations suggest that OCLs and macrophages have different phagosome maturation mechanisms that involve the specific and regulated acquisition of markers from endocytic organelles. The results also demonstrate that the use of LBC is a useful system in which to identify and characterize molecules involved in these different endocytic pathways. PMID- 11726284 TI - Identification and characterization of CaMKP-N, nuclear calmodulin-dependent protein kinase phosphatase. AB - Calmodulin-dependent protein kinase phosphatase (CaMKP) dephosphorylates and concomitantly deactivates multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaMKs), such as CaMKI, CaMKII, and CaMKIV. In the present study, a nuclear CaMKP-related protein, CaMKP-N, was identified. This protein consisted of 757 amino acid residues with a calculated molecular weight of 84,176. Recombinant CaMKP-N dephosphorylated CaMKIV. The activity of CaMKP-N requires Mn(2+) ions and is stimulated by polycations. Transiently expressed CaMKP-N in COS-7 cells was localized in the nucleus. This finding together with previous reports regarding localization of CaMKs indicates that CaMKP-N dephosphorylates CaMKIV and nuclear CaMKII, whereas CaMKP dephosphorylates CaMKI and cytosolic CaMKII. PMID- 11726285 TI - Three soluble form messages of murine CD46 are produced through alternative mRNA splicing. AB - Murine CD46 (mCD46) is a type 1 membrane protein expressed predominantly in testicular germ cells, the distribution profile of which is in contrast to that of human CD46 showing a ubiquitous tissue distribution. We have identified an additional message of mCD46 that encodes a putative secretory form [Nomura et al. (1999) Immunogenetics 50, 245-254]. Here, we cloned three cDNAs encoding putative soluble CD46 from murine testis. These soluble form messages were yielded on insertion of unidentified nucleotide sequences, 77, 179, and 73 ntds, into the junctions between the SCR3 and SCR4 (variant 2), ST(c) and UK (variant 3), and SCR4 and ST(c) (variant 1) domains, respectively, the last one corresponding to the reported soluble form. The exons corresponding to these three inserts were identified in the murine CD46 genome, indicating that the alternative splicing of mRNA participates in the generation of these various CD46 messages. In normal mouse sera and cell lines, however, virtually no soluble CD46 was detected on immunoblotting. On Northern blotting analysis with specific probes, on the other hand, variant 1 was found to be predominantly expressed in the liver and heart. In addition, all variant messages were detected on PCR in all organs examined. When a rabbit cell line, RK13 cells, was transfected with cDNA of variant 1, protein synthesis was detected on immunoblotting. Although the mCD46 protein production was inefficient, this variant 1 exhibited factor I-cofactor activity as to inhibition of the complement cascade. Since the mCD46 protein was reported to be markedly up-regulated on infection of murine cells with mCMV, the soluble mCD46 proteins may act as a complement regulator that controls the systemic complement system under the conditions of a viral infection. PMID- 11726286 TI - Gonadotropin-dependent expression of sterol 14-demethylase P450 (CYP51) in rat ovaries and its contribution to the production of a meiosis-activating steroid. AB - Immunohistochemical analysis showed that sterol 14-demethylase P450 (CYP51) is expressed in mature follicles and corpus lutea of rat ovaries. In follicles, CYP51 is expressed in granulosa and theca cells but not in oocytes. The ovarian CYP51 activity of hypophysectomized rats is very low and induced by pregnant mares' serum gonadotropin (PMSG) treatment together with ovarian growth. The expression of CYP51 first increases in growing follicles and then appears in the corpus lutea after luteinization. The former event may be due to the follicular stimulating hormone action of PMSG, and the latter may be caused by the luteinizing hormone effect of PMSG. Sterol analysis indicated that the product of the CYP51-mediated lanosterol 14-demethylation, 4,4-dimethylcholesta-8,14,24 trienol, which has been identified as a meiosis-activating steroid (MAS) in mammals [Byskov et al. (1995) Nature 374, 559-562], accumulates (about 10 pmol/mg of ovary) in mature rat ovaries, and the content is enough to activate the resumption of meiosis. These lines of evidence suggest that the expression of ovarian CYP51 is dependent on gonadotropins, and ovarian CYP51 activity is enough for accumulating MAS. Serum insulin does not affect the ovarian CYP51 level, although it is essential for hepatic CYP51 expression. These findings indicate that expression of CYP51 is regulated differently among organs. PMID- 11726287 TI - Bombyx cysteine proteinase inhibitor (BCPI) homologous to propeptide regions of cysteine proteinases is a strong, selective inhibitor of cathepsin L-like cysteine proteinases. AB - Bombyx cysteine proteinase inhibitor (BCPI) is a novel cysteine proteinase inhibitor. The protein sequence is homologous to the proregions of certain cysteine proteinases. Here we report the mechanism of its inhibition of several cysteine proteinases. BCPI strongly inhibited Bombyx cysteine proteinase (BCP) activity with a K(i) = 5.9 pM, and human cathepsin L with a K(i) = 36 pM. The inhibition obeyed slow-binding kinetics. The inhibition of cathepsin H was much weaker (K(i) = 82 nM), while inhibition of papain (K(i) > 1 microM) and cathepsin B (K(i) > 4 microM) was negligible. Following incubation with BCP, BCPI was first truncated at the C-terminal end, and then gradually degraded over time. The truncation mainly involved two C-terminal amino acid residues. Recombinant BCPI lacking the two C-terminal amino acid residues still retained substantial inhibitory activity. Our results indicate that BCPI is a stable and highly selective inhibitor of cathepsin L-like cysteine proteinases. PMID- 11726288 TI - Energetic feasibility of hydrogen abstraction and recombination in coenzyme B(12) dependent diol dehydratase reaction. AB - Coenzyme B(12) serves as a cofactor for enzymatic radical reactions. The essential steps in all the coenzyme B(12)-dependent rearrangements are two hydrogen abstraction steps: hydrogen abstraction of the adenosyl radical from substrates, and hydrogen back-abstraction (recombination) of a product-derived radical from 5'-deoxyadenosine. The energetic feasibility of these hydrogen abstraction steps in the diol dehyratase reaction was examined by theoretical calculations with a protein-free, simplified model at the B3LYP/6-311G* level of density functional theory. Activation energies for the hydrogen abstraction and recombination with 1,2-propanediol as substrate are 9.0 and 15.1 kcal/mol, respectively, and essentially not affected by coordination of the substrate and the radical intermediate to K+. Since these energies can be considered to be supplied by the substrate-binding energy, the computational results with this simplified model indicate that the hydrogen abstraction and recombination in the coenzyme B(12)-dependent diol dehydratase reaction are energetically feasible. PMID- 11726289 TI - Deletion of mitochondrial ATPase inhibitor in the yeast Saccharomyces cerevisiae decreased cellular and mitochondrial ATP levels under non-nutritional conditions and induced a respiration-deficient cell-type. AB - T(1), a mutant yeast lacking three regulatory proteins of F(1)F(o)ATPase, namely ATPase inhibitor, 9K protein and 15K protein, grew on non-fermentable carbon source at the same rate as normal cells but was less viable when incubated in water. During the incubation, the cellular ATP content decreased rapidly in the T(1) cells but not in normal cells, and respiration-deficient cells appeared among the T(1) cells. The same mutation was also induced in D26 cells lacking only the ATPase inhibitor. Overexpression of the ATPase inhibitor in YC63 cells, which were derived from the D26 strain harboring an expression vector containing the gene of the ATPase inhibitor, prevented the decrease of cellular ATP level and the mutation. Isolated T(1) mitochondria exhibited ATP hydrolysis for maintenance of membrane potential when antimycin A was added to the mitochondrial suspension, while normal and YC63 mitochondria continued to show low hydrolytic activity and low membrane potential. Thus, it is likely that deletion of the ATPase inhibitor induces ATPase activity of F(1)F(o)ATPase to create a dispensable membrane potential under the non-nutritional conditions and that this depletes mitochondrial and cellular ATP. The depletion of mitochondrial ATP in turn leads to occurrence of aberrant DNA in mitochondria. PMID- 11726290 TI - Cloning and characterization of motX, a Vibrio alginolyticus sodium-driven flagellar motor gene. AB - Four motor proteins, MotX, MotY, PomA, and PomB, have been identified as constituents of the Na(+)-driven flagellum of Vibrio species. In this study, the complete motX gene was cloned from Vibrio alginolyticus and shown to complement three mot mutations, motX94, motX115, and motX119, as well as a V. parahaemolyticus motX mutant. The motX94 mutant contains a frameshift at Val86 of MotX, while the motX115 and motX119 mutations comprise substitutions of Ala146 to Val and Gln 194 to amber, respectively. When MotX was overexpressed in Vibrio cells, the amount of MotY detected in the membrane fraction increased, and vice versa, suggesting that MotX and MotY mutually stabilize each other by interacting at the membrane level. When a plasmid containing the motX gene was introduced into motY mutants NMB117 (motY117) and VIO542 (motY542), the mutations were suppressed. In contrast, motY could not cause the recovery of any swarm-defective motX mutants studied. Considering the above evidence, we propose that MotX is more directly involved than MotY in the mechanical functioning of the Na(+)-type flagellar motor, and that MotY may stabilize MotX to support its interaction with other Mot proteins. PMID- 11726291 TI - Sp family members stimulate transcription of the hex gene via interactions with GC boxes. AB - The 5'-flanking region of the mouse Hex gene was examined in order to identify transcription factors regulating its expression in hepatocytes and haematopoietic cells. We have identified two further GC boxes (GC boxes 3 and 4 at nucleotide positions -149 to -140 and -79 to -70, respectively), i.e. in addition to the two previously determined ones (GC boxes 1 and 2 at nucleotide positions -197 to -188 and -176 to -167, respectively). Luciferase reporter assays revealed that all four GC boxes are transcriptionally active in both MH(1)C(1) rat hepatoma and K562 human chronic myelogenous leukemia cells. Electrophoretic mobility shift assays with specific competitors and antibodies showed that members of the Sp family, namely Sp1 and Sp3, bind to these GC boxes. Overexpression of Sp1 and Sp3 in Drosophila SL2 cells stimulated transcription of the Hex gene through interactions with GC boxes 1 to 4, Sp1 being a more potent activator than Sp3. Thus, we conclude that Sp1 and Sp3 stimulate transcription of the Hex gene in both MH(1)C(1) and K562 cells. PMID- 11726292 TI - Evaluation of emergency air evacuation of critically ill patients from cruise ships. AB - BACKGROUND: The study objectives were to assess the ship physician's diagnostic accuracy in making the decision to air evacuate critically ill patients from cruise ships, to determine the outcome of these patients, and the overall benefit of air evacuation. METHODS: From October 1999 to May 2000, we performed a prospective study of critically ill patients coming from cruise ships in the Caribbean and transported to our institution by air ambulance. Demographics, initial diagnosis, and treatment on board were collected by the triage officer at the time of the cruise physician's first call. In route complications and flight team composition were obtained from the air ambulance monitoring log. Patients were followed-up in the hospital for complications, outcome, and final diagnosis. RESULTS: A consecutive series of 104 patients were considered for analysis. There were 65 men and 39 women (mean age: 68.7 years). Cruise physician's diagnosis was correct in more than 90% of the cases. Internal medicine and surgical conditions represented 80.8% and 19.2% of the cases respectively, falling mainly into three categories: cardiac (34.6%), neurological (20.2%), and digestive (14%). Two cardiac arrests and 1 ventricular fibrillation were successfully resuscitated and 5 of 15 myocardial infarctions received thrombolytic therapy on board. Air transfers were warranted in 96.1% of the cases and physician presence in the flight was considered appropriate in 97.6%. In route complications and mortality rate were 5.8% and 2.9% respectively, related to serious cardiac events. Among the 98 hospitalized patients, 10 patients developed new complications and 5 died. The overall mortality rate was 7.7%. CONCLUSION: The cruise industry appears off to a good start in the medical treatment of passengers needing air evacuation to a land based medical facility. There is room for improvement and adoption of American College of Emergency Physicians (ACEP) and International Council of Cruise Lines (ICCL) Health Care Guidelines are meaningful first steps. Analysis of Caribbean medical facilities and implementation of active telemedicine conferencing represent alternatives to air evacuation that need to be studied. PMID- 11726293 TI - Typhoid and paratyphoid fever: a 10-year retrospective study of 41 cases in a Parisian hospital. AB - BACKGROUND: Enteric fever remains a major cause of fever in travelers. We evaluated new trends in enteric fever. METHODS: We reviewed the epidemiological, clinical, biological, bacteriological data, and outcome of all cases of typhoid and paratyphoid fever seen in our department over the last decade. The inclusion criteria were the presence of signs compatible with enteric fever and isolation of Salmonella typhi or Salmonella paratyphi A, B, or C from blood or stool cultures or any other site. RESULTS: Among the 41 patients, 38 (93%) had travel associated enteric fever. The main geographic source of contamination was the Indian subcontinent. One patient had been vaccinated with parenteral Vi vaccine 1 year previously. Fever and headaches were the only signs which were present in more than 80% of patients. The Widal test at inclusion was positive in 27%, and a second serological test was found to be positive in 50% of evaluated cases. Blood cultures and stool cultures were positive in 34 cases and 10 cases, respectively. Salmonellae spp were isolated in both hemocultures and stool cultures in 4 cases and in urine in 1 case. Two strains of S. typhi were resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole. One strain of S. typhi and one of S. paratyphi B were nalidixic acid resistant. All evaluable patients were cured with the exception of 2 patients (1 failure, 1 relapse). We observed 3 toxic reactions. No patients died. CONCLUSION: The diagnosis and outcome of enteric fever are hampered by the lack of specificity of clinical and biological signs, the increasing rates of antimicrobial resistance, and the occurrence of toxic reactions during treatment. PMID- 11726294 TI - Knowledge of malaria, risk perception, and compliance with prophylaxis and personal and environmental preventive measures in travelers exiting Zimbabwe from Harare and Victoria Falls International airport. AB - BACKGROUND: Travel associated malaria is a major health risk for visitors to malaria endemic destinations. To examine the knowledge of malaria prevention, risk perception, current prophylactic behavior, and compliance with chemoprophylaxis and personal and environmental protection measures we conducted a study in a cohort of travelers exiting Zimbabwe from two international airports during a peak malaria transmission period. METHODS: Data were collected by pretested self-administered questionnaires from 595 adults in the departure lounges of Harare and Victoria Falls International airports. Excluded were children and travelers from the African continent. A multilingual research assistant supervised data collection. RESULTS: The majority of travelers obtained health advice prior to travel. Patterns of protective behavior and compliance with prophylaxis were inconsistent with a high perception of malaria threat and good knowledge. About 23% of travelers failed to use chemoprophylaxis during their visit. In the group of travelers who used chemoprophylaxis, 18% were noncompliant. Fifteen drug combinations were in use. Full compliance with medication plus use of personal preventive measures always was estimated as 13%. Forgetfulness was the main cause of noncompliance, followed by deliberate omission due to side effects. Of 57 travelers who reported side effects from current medication, over half used mefloquine. CONCLUSIONS: There is a need to examine how people process personal risk and communications about risk. We must recognize the competition between precautionary measures against malaria and other life demands that are imposed by travel, especially in young long stay travelers and persons visiting primarily for business purposes. Mediating a protective response will also depend on judgments about the effectiveness of the action, strengthening travelers intentions toward adherence, and increasing efficacy perception by individuals and their peers. Conflicts in prophylactic recommendations need to be resolved. As ecotourism develops in Zimbabwe and other malaria regions, stakeholders in this rapidly growing industry must be made aware of the important role they can play in protecting clients from malaria. PMID- 11726296 TI - Sailing safely. PMID- 11726295 TI - Preventing infectious diseases in long-term travelers to rural Africa. PMID- 11726297 TI - Acute bilharziasis outbreak in a family visiting Mali. PMID- 11726298 TI - Hyperreactive malarial splenomegaly in Europeans: report of five cases. PMID- 11726299 TI - Multisystem Schistosoma haematobium infection in an Australian tourist. PMID- 11726300 TI - Italian travelers to Asia: a report on two cases of severe hepatitis a infection evolving to complete recovery. PMID- 11726301 TI - Imported malaria in pregnancy due to Plasmodium falciparum. PMID- 11726302 TI - Selected bibliography. PMID- 11726304 TI - Cardiovascular effects of sildenafil citrate (Viagra): a naturalistic cross-over study. AB - In a cross-over study we investigated the effects of sildenafil (single doses of 25- or 50mg) on cardiovascular autonomic nervous system (ANS) function assessed by serial recordings of blood pressure, conventional 12-lead electrocardiograms and standardized, time-and frequency domain indices of heart rate variability (HRV) in 21 men with erectile dysfunction. More than half of these patients had multiple comorbidities. Sildenafil induced significant mean reductions from baseline in resting blood pressure, accompanied by a reflex increase in heart rate. There were no significant changes after administration of sildenafil in any other of the ANS function indices. These preliminary findings suggest that sildenafil does not significantly affect cardiac ANS function in patients with erectile dysfunction. PMID- 11726305 TI - Differential effects of radiocontrast agents on human umbilical vein endothelial cells: cytotoxicity and modulators of thrombogenicity. AB - The endothelium plays a central role in the regulation of blood flow and coagulation. The impact of radiocontrast agents on endothelial cells is therefore potentially clinically important, particularly in percutaneous interventions for acute coronary thrombosis. The effects of radiocontrast agents on endothelial cell viability and determinants of thrombogenicity were studied in human umbilical vein endothelial cells (HUVECs) in vitro. Intercellular tight junctions were assessed using immunofluorescence microscopy and measurement of the transmonolayer electrical resistance (TMR). The concentrations of endothelin-1 (E), von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1) and thrombomodulin (T) were measured in the cell culture media. The ionic, high osmolal radiocontrast agent diatrizoate induced concentration-dependent cell death and an opening of tight junctions with the attendant abolition of the TMR. The concentration of E decreased, vWF increased in the cell culture media, the concentration of PAI-1 and T was not significantly changed by diatrizoate. Radiocontrast agents with reduced osmolality (ioxaglate: ionic; iopamidol: non ionic) induced an increase in PAI-1 and vWF, but E and T were not significantly changed. CONCLUSIONS: Radiocontrast agents have differential effects on endothelial cells in vitro including the secretion of modulators of thrombogenesis. The effects are most pronounced in the markedly hyperosmolal compound diatrizoate suggesting a contributory role of hypertonicity. Ioxaglate and iopamidol both increased the prothrombotic factors vWF and PAI-1 to the same degree indicating a similar risk of thrombogenicity between low-osmolal ionic and non-ionic radiocontrast agents in this in vitro model. PMID- 11726306 TI - Sixteen rare sequence variants of the hMLH1 and hMSH2 genes found in a cohort of 254 suspected HNPCC (hereditary non-polyposis colorectal cancer) patients: mutations or polymorphisms? AB - 5-8% of all colorectal cancer cases are assumed to be due to germline mutations in DNA mismatch repair genes. Mutation analysis of these genes in affected families enables one to identify subjects with an inborn susceptibility to colorectal tumorogenesis and to offer presymptomatic testing to family members at risk, provided that the mutation detected is a truncating one or a missense mutation that has either been judged as disease causing in other families or segregates with the disease and results in a microsatellite instability of the corresponding tumor. Segregation analysis within the family or microsatellite analysis of the tumor is, however, not always possible. In these cases, assessment of the relevance of the sequence variation identified is very difficult. On the other hand, discrimination between inactivating mutations and innocuous sequence polymorphisms is of extreme importance for clinical and genetic counseling of affected families. Here we report 16 rare sequence variants of the hMLH1 and hMSH2 genes including 11 different missense variations found in a cohort of 254 suspected HNPCC patients. We provide evidence, that missense variations in hMLH1 do not necessarily result in microsatellite instability of the corresponding tumor DNA. These patients would have been missed had one followed the recommendations of using only microsatellite analysis for the selection of patients at high risk of hereditary non-polyposis colorectal cancer for mutation analysis. PMID- 11726307 TI - Clinical effectiveness of a potassium chloride containing chewing gum in the treatment of hypersensitive teeth. AB - Dental hypersensitivity has been observed in an increasing number of younger patients over the past few decades. The reasons for this include forced or false tooth-brushing techniques, and abrasions caused by bruxism or orthodontic procedures. The aim of the present study was to assess the possible benefits of a potassium chloride (KCl) containing chewing gum in dental hypersensitivity. The study population consisted of 59 selected university students (age: 25 +/- 4.4 years). Each subject suitable for entry into the study had more than 3 teeth sensitive to thermal stimulation (air stimulus range of 30-80 mm on a 100mm visual analogue scale (VAS), and showed signs of facial/cervical erosion, abrasion and/or gingival recession. Exclusion criteria were daily doses of medication, teeth with deep restorations, a gingival score of > 2, or periodontal surgery within the past 6 months. The total number of hypersensitive teeth was 217. Over a period of 3 weeks the subjects brushed with a non-desensitising toothpaste and, in addition, used the KCl chewing gum up to 6 times daily for 10 min on each occasion. This was followed by a period of 3 weeks of non desensitising toothpaste use alone. The subjects were questioned on the severity of intraoral pain and asked to indicate the intensity of the pain on the VAS (baseline, 1, 3 and 6 weeks). Mean thermal sensitivity at baseline was 54.5 +/- 9 mm. There was a drop to 40.8 mm ( 1.9) after 1 week and to 28.6 mm (+/- 2) after 3 weeks. The reduction was statistically significant (p <0.001) at both measurement points. The mean rating increased to 29.1 mm after 6 weeks. The results of this study demonstrate that the daily use of a KCl containing chewing gum is a useful non-invasive method of reducing dental hypersensitivity over an extended period of time. PMID- 11726308 TI - Uptake of oxygen from the intestine-- experiments with rabbits. AB - BACKGROUND: Venous portal blood contributes essentially to the oxygen supply of the liver. The effect of oxygen enriched water, applied into the stomach, on oxygenation of portal blood was investigated in rabbits. MATERIAL AND METHODS: 15 anaesthetized rabbits were given 30 ml water, containing 45, 80 or 150 mg O2/l, by a gastric tube. Oxygen pressure was monitored continuously by a measuring probe in the stomach, the abdominal cavity and in the gastric as well as in the portal vein. RESULTS: Intragastrically applied water delivered oxygen slowly, according to the increase in its temperature. The released oxygen was found to penetrate into the abdominal cavity, and a dose-response relationship between oxygen pressure in the stomach and the abdominal cavity was established. 45 mg O2/l water resulted in a negligible increase of oxygen concentration in the abdomen, while 80 or 150 mg O2/l lead to an increase of oxygen by 10 rsp. 20 mmHg in the abdomen, and by up to 14 mmHg in the portal vein. Oxygen penetration occurred according to the known physical and physiological parameters of gas diffusion. Diffusion of oxygen was enhanced, if concurrently with oxygen the water was enriched with CO2. CONCLUSIONS: Our results show that intragastrically applied oxygenated water with more than 45 mg O2/l delivers O2 into the abdominal cavity and the portal vein. This effect may be of clinical relevance in states of impaired liver perfusion such as fatty liver or hepatitis. PMID- 11726310 TI - Sulfasalazine-induced extrinsic allergic alveolitis in a patient with psoriatic arthritis. AB - We report the first case of a well defined extrinsic allergic alveolitis as a complication of sulfasalazine therapy in a patient treated for psoriatic arthritis. CT of the chest showed small nodular densities over both lungs, BAL demonstrated a highly active lymphocytic alveolitis and transbronchial biopsies revealed lymphoplasmocytic interstitial infiltration. Sulfasalazine as causative agent was proven by an inadvertent rechallenge three years later and a positive lymphocyte transformation test. sulfasalazine; psoriatic arthritis; extrinsic allergic alveolitis PMID- 11726311 TI - IgG antibodies typical for extrinsic allergic alveolitis--an inter-laboratory quality assessment. AB - BACKGROUND: Determination of specific IgG antibodies is important for the diagnosis of extrinsic allergic alveolitis (EAA). Various evaluations have however shown, that current methodology lacks sufficient standardization in that the employment of different sources of extracts and techniques makes a comparison of data from one laboratory to another almost impossible. OBJECTIVE: The aim of this study is to establish an external quality control system and to analyse, what the explanations for the different outcomes from various laboratories might be. METHODS: In the past 4 years 5 sera from patients suffering from EAA or healthy controls were sent every 6 months to 11 different allergy laboratories in Austria. The determination of specific IgG antibodies against antigens that are typical for this disease were requested. Results were gained with the method routinely used in the respective laboratory, and then sent back to the reference center for statistical evaluation. Precipitating techniques were used in 8 laboratories during the first mailings, but were gradually exchanged by automated ELISA systems being employed in 8 laboratories in the last mailing. RESULTS: 1127 values were determined in 105 expectedly positive sera and 1003 in 94 negative samples. Of the 562 values obtained with precipitation techniques in positive sera, only 52.0% were reported to be positive, and the results varied considerably among laboratories and antigens. In contrast, 93.3% were positive with commercially available ELISA techniques, with 92.3% for the EnzyDex System and even 95.5% for the UniCAP System. Regarding the specificity however, 93.0% of the expected negative results were correct negative using precipitation methods, whereas merely 75.2% were negative with the EnzyDex System and only 22.5% using the UniCAP System. Moreover 35.8% of the results using this latter method were false-positive. CONCLUSIONS: The traditional precipitation techniques proved not only technically difficult to perform, but also unreliable, difficult to reproduce, insensitive and impractical in daily laboratory work. They suffer from that many draw backs, that their use in daily routine cannot be recommended any more. Automated ELISA systems seem to fulfill the criteria for a routine technique concerning handling, automation, and quality criteria like sensitivity quite well, but not for specificity. Both techniques urgently need external standardization in order to make the results comparable among the different systems and methods; the danger of potentially false-positive results, pretending sensitizations that might be clinically irrelevant in several cases, is high. PMID- 11726312 TI - Inside Haematologica: new tools towards risk classification according to genetic lesions in acute lymphoblastic leukemia. PMID- 11726313 TI - An Italian pioneer in the study of tumor angiogenesis. PMID- 11726314 TI - Blood cells diseases and thrombosis. AB - BACKGROUND AND OBJECTIVES: In recent years knowledge about thrombophilia and the mechanisms underlying the pathogenesis of thrombosis has increased greatly. Nevertheless the role of leukocytes and red cells in thrombogenesis is not well established and is probably underestimated. EVIDENCE AND INFORMATION SOURCES: The contribution of leukocytes and red cells to thrombogenesis has been reviewed. Moreover, the prevalence of thrombosis as a complication of hematologic diseases has been examined. The authors are involved in the investigation and management of acute and chronic hematologic diseases as well as in investigation of thrombophilia. Pub-Med was employed as a source of information. STATE OF THE ART: Thrombosis is a major problem in myeloproliferative disorders such as polycythemia vera and essential thrombocythemia. A clonal involvement of megakaryocytopoiesis resulting in elevated levels of platelet-specific proteins, increased thromboxane generation, and expression of activation-dependent epitopes on the platelet surface is regarded as the main origin of thromboembolism; nevertheless, activation of leukocytes and the consequent release of elastase and alkaline phosphatase could play an important role, determining endothelial damage. Thrombosis is a relevant problem in some hemolytic anemias such as paroxysmal nocturnal hemoglobinuria and drepanocytosis. Thrombotic events in hemolytic anemias with membrane defects have been attributed, at least in part, to hypercoagulability related to the exposure of phosphatidylserine of red cell membrane activating plasma prothrombinase and supplying a procoagulant phospholipid anionic surface. A moderate but well-established risk for thrombosis occurs in acute promyelocytic leukemia and acute lymphoblastic leukemia; this risk could be increased by antiblastic drugs affecting the procoagulant activity of cells and the production of coagulation inhibitors from the liver. PERSPECTIVES: Thrombotic complications during hematologic diseases other than thrombophilia due to plasma alteration could be decreased not only by anticoagulant and antiaggregating agents but also by drugs inhibiting activation of leukocytes and red cells and their interaction with platelets. PMID- 11726315 TI - Fluorescence in situ hybridization study of TEL/AML1 fusion and other abnormalities involving TEL and AML1 genes. Correlation with cytogenetic findings and prognostic value in children with acute lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: The TEL/AML1 fusion is the most common genetic abnormality found in childhood acute lymphoblastic leukemias (ALL). Although it is very difficult to identify by conventional cytogenetic techniques it can be readily detected using fluorescence in situ hybridization (FISH). We carried out cytogenetic and FISH studies on 42 children with ALL in order to know the frequency of this translocation in our population, the incidence of TEL and/or AML1 gene alterations, and their correlation with clinical evolution and prognosis. In addition, we performed reverse transcription polymerase chain reaction (RT-PCR) in some cases, confirming the feasibility of FISH techniques in the detection of this translocation. DESIGN AND METHODS: Bone marrow samples were obtained from 42 childhood ALL patients. The copy number of AML1 and TEL genes were studied using fluorescent in situ hybridization with a dual color DNA probe specific for the AML1 and TEL genes. RESULTS: We found a frequency of TEL/AML1 fusion of 17% in our sample. Double TEL/AML1 fusion, lack of TEL signal and extra AML1 signals were frequent additional FISH abnormalities. Duplication of a chromosomal complement, deletion of chromosome 12p arm, and polysomies of chromosome 21 are plausible explanations for these additional FISH findings. However, a relatively high proportion of our cases (9.5%) presented specific amplification of AML1. A statistically significant difference in prognosis was found between patients with and without these additional AML1 or TEL FISH alterations (p<0.02), which could be related to the presence of specific karyotypes. INTERPRETATIONS AND CONCLUSIONS: The frequency of TEL/AML1 fusion is similar to that found in other populations (17%). We found that FISH analysis of AML and TEL is related to the evolution of the disease. The absence of alterations in these genes revealed by FISH could be indicative of bad prognosis, while the presence of alterations is related to a good evolution. Our results suggest that interphase FISH analysis to search for alterations in AML and TEL genes could be extremely useful for complementing cytogenetic studies and for providing additional information about the possible outcome of the disease in patients with ALL. PMID- 11726316 TI - Multiplex-polymerase chain reaction assay for the detection of prognostically significant translocations in acute lymphoblastic leukemia. AB - BACKGROUND AND OBJECTIVES: The presence of specific chromosomal translocations in acute lymphoblastic leukemias (ALL) plays an important role in determining the prognosis of the patients. Our aim is to develop a highly sensitive and specific method to screen simultaneously for the four most frequent translocations in ALL: t(9;22), t(1;19), t(4;11), t(12;21). DESIGN AND METHODS: Our approach uses a multiplex-polymerase chain reaction (PCR) method, which involves two rounds of PCR using fluorescence-labeled nested primers. The chimeric transcripts resulting from these translocations can be identified by agarose gel electrophoresis or by fluorescence analysis. To validate this method we carried out the analysis in 42 pediatric ALL samples previously studied by cytogenetic and fluorescent in situ hybridization (FISH) techniques. RESULTS: In all samples with a known translocation detected by cytogenetic or FISH techniques, the same translocation was identified by the multiplex-PCR assay. Moreover, with this method we detected rearrangements in five patients in clinical remission and in two patients at diagnosis for whom karyotypes were normal and rearrangements had not been detected. The application of this multiplex-PCR assay was also useful in cases without cytogenetic results. INTERPRETATION AND CONCLUSIONS: These results show that the multiplex-PCR method allows reliable, sensitive and rapid detection of the prognostically significant translocations in ALL. We believe that this assay combined with cytogenetic analysis should be the strategy of choice for the initial diagnostic phase of acute lymphoblastic leukemia, and that it could be used not only at diagnosis but also to follow-up these alterations in remission samples without previous controls. PMID- 11726317 TI - Interleukin-3 receptor alpha chain (CD123) is widely expressed in hematologic malignancies. AB - BACKGROUND AND OBJECTIVES: The hematopoietic system is controlled by growth factors (cytokines) which can influence cell survival, proliferation, differentiation and functional activation. The study of cytokine receptor expression by flow cytometry could allow us to differentiate between normal and tumoral cells. DESIGN AND METHODS: We analyzed the expression of the interleukin 3 (IL-3) receptor a chain (CD123) in 22 normal samples and in a wide panel of hematologic malignancies using flow cytometry. We found that CD123 was expressed in the myeloid progenitor subpopulation but in contrast, normal lymphoid progenitors lacked CD123. We analyzed the CD123 expression pattern in 64 patients with acute leukemia, 45 with acute myeloid leukemia (AML) and 19 with acute lymphocytic leukemia (ALL) (13 B-cell lineage ALL and 6 T-cell lineage ALL). RESULTS: All the AML cases except two patients with M7 and all the B-cell lineage ALL patients were CD123 positive. In contrast, all the T-cell lineage ALL cases tested were CD123 negative. We also studied the CD123 expression pattern in 122 patients with a B-cell chronic lymphoproliferative disease (B-CLPD). CD123 was positive in three situations: 1) typical cases of hairy cell leukemia showed a specific, strong CD123 expression, 2) a subgroup of atypical chronic lymphocytic leukemia with a marked CD11c expression was also CD123 positive, and finally 3) transformed B-CLPD showed the phenomenon of transition from CD123 negativity to CD123 positivity simultaneuosly with morphologic changes. INTERPRETATION AND CONCLUSIONS: In summary, our data show high expression of IL-3 receptor a chain in hematologic malignancies. Given the high frequency of CD123 reactivity in blast cells in contrast to in normal precursors, this antigen could be applied to the study of minimal residual disease in acute leukemia. CD123 is expressed with a characteristic pattern in cases of hairy cell leukemia. PMID- 11726318 TI - Fetal and embryonic hemoglobins in erythroblasts from fetal blood and fetal cells enriched from maternal blood in fetal anemia. AB - BACKGROUND AND OBJECTIVES: To determine whether there is a delay or reversal in switch mechanisms from embryonic (e and z) to fetal (g) hemoglobins accompanying the erythroblastosis of anemic fetuses and whether an increased erythroblast count in fetal blood is associated with an increase in feto-maternal cell trafficking. DESIGN AND METHODS: Fetal and maternal blood samples were obtained from 10 cases with rhesus isoimmunization and 2 cases with maternal Parvo-B19 virus at 19-33 weeks' gestation. Blood samples were also taken as controls from 61 fetuses and 86 mothers. Fetal erythroblasts were isolated by triple density gradient centrifugation and magnetic cell sorting with CD71 antibody. Fluorescent antibodies were used to immuno-stain for zeta (z), epsilon (e) and gamma (g) hemoglobin chains. In the maternal samples, fluorescence in situ hybridization (FISH) for X and Y chromosomes was also carried out to confirm the presence and proportion of the enriched fetal cells from maternal blood. RESULTS: In both fetal and maternal blood the percentage of erythroblasts positive for g-globin chain was significantly higher in the anemic fetuses compared to the controls (fetal blood, p<0.001, R=0.91; maternal blood, p<0.001, R=0.56), but there was no significant difference in expression of the e and z-chains. The percentage of cells with Y-signals was also higher in the maternal samples of anemic fetuses compared to normal controls (p<0.001, R=0.56). INTERPRETATION AND CONCLUSIONS: These findings suggest that the erythroblastosis of anemic fetuses is not accompanied by a delay or a reversal in switch from embryonic to fetal hemoglobin chains. Severe fetal anemia is associated with an increase in feto-maternal cell trafficking. PMID- 11726319 TI - Spontaneous remission in adult patients with de novo myelodysplastic syndrome: a possible event. AB - BACKGROUND AND OBJECTIVES: Spontaneous remission (SR) in de novo myelodysplastic syndromes (MDS) is a rare event, which has been so far described only in children with monosomy 7. The phenomenon is extremely heterogeneous, perhaps depending on different pathogeneses of the disease. DESIGN AND METHODS: We retrospectively evaluated the outcome of 564 consecutive adult patients with primary MDS diagnosed at our Institution in a 12-year period. SR was defined as an unexpected improvement lasting more than 1 year without concomitant treatments other than vitamins or low-dose steroids (in patients with platelets < 50 x 10(9)/L). RESULTS: Nine cases of SR were observed in 3 males and 6 females (median age 38.7 years). At diagnosis, all patients had Hb levels < 10 g/dL and 8/9 required packed red cell transfusions. The median time from diagnosis to SR was 18 months (range 4-46) and all patients had normalization of peripheral blood parameters: in 2 out of 3 patients with karyotypic abnormalities at onset, a cytogenetic remission was documented. The median duration of SR was 56 months; 5 patients are still in SR and 4 patients have relapsed (1 as MDS and 3 as acute myeloid leukemia). INTERPRETATION AND CONCLUSIONS: SR is a rare (less than 2% in our experience) but possible event also in adult MDS patients. It should be kept in mind in the evaluation of experimental treatments for MDS in which very low rates of complete responses are expected. PMID- 11726320 TI - Interferon alpha plus intermittent oral Ara-C ocfosfate (YNK-01) in chronic myeloid leukemia primarily resistant or with minimal cytogenetic response to interferon. AB - BACKGROUND AND OBJECTIVES: Subcutaneous Ara-C plus interferon (IFN) produces more cytogenetic responses than IFN in chronic myeloid leukemia (CML) but a greater toxicity. The objective of this study was to determine the efficacy and tolerance of IFN plus oral Ara-C ocfosfate (YNK-01) in IFN-resistant CML patients. DESIGN AND METHODS: A phase II pilot study was conducted in 19 CML patients primarily resistant or with minimal cytogenetic response to IFN. Patients were scheduled to receive 6 monthly 14-day cycles of YNK-01 (500 mg/day), with progressive escalation if tolerated, in addition to IFN. Cytogenetic assessment was performed thereafter. RESULTS: Of the first 7 patients, 5 had severe hematologic and 5 moderate gastrointestinal toxicity; IFN was reduced in 6, YNK-01 in 5, and treatment discontinued in 2; hematologic response was achieved in 2 of the 5 evaluable patients. In the following 4 patients the Ara-C was reduced to 300 mg: 2 had severe hematologic and 2 moderate gastrointestinal toxicity; IFN and Ara-C were reduced in 2 patients and treatment discontinued in 2 due to progression or toxicity; the other 2 achieved a minor cytogenetic response, progressing in one to a major response after 6 more cycles. In 8 patients the starting Ara-C dose was 200 mg: 5 had moderate-severe hematologic and 5 mild gastrointestinal toxicity; IFN was reduced in 5, Ara-C in 1, and treatment discontinued in 1; Ara C was increased in 7 cases; hematologic response was obtained in 4 patients, 2 of whom attained a minor and 1 a major cytogenetic response. INTERPRETATION AND CONCLUSIONS: These results provide background for future studies aimed at ascertaining the role of oral Ara-C combined with IFN or STI571 in newly diagnosed CML. PMID- 11726321 TI - Multiparametric analysis of apoptotic and multi-drug resistance phenotypes according to the blast cell maturation stage in elderly patients with acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Acute myeloid leukemia (AML) is a heterogeneous group of malignant diseases, often characterized by coexistence of more than one subpopulation of blast cells. Multiparametric flow cytometry immunophenotyping has proven to be a reliable and sensitive approach for the discrimination of myeloid blast cells from residual normal cells present in bone marrow samples from AML patients and, at the same time, allows the identification of different maturation compartments among myeloid blasts. Therefore, it provides a unique tool for assessing apoptotic and multidrug resistance (MDR)-associated phenotypes in individual subsets of leukemic cells. DESIGN AND METHODS: The aim of the present study was to explore the simultaneous expression of proteins related to both apoptosis (APO2.7, bcl-2, bax) and multidrug resistance (MDR1, MRP, LRP) in the different blast cell subpopulations detected at diagnosis in a group of 72 elderly patients with AML. In addition, we included 5 bone marrow samples from healthy adult donors in the analysis. RESULTS: Immature blast cells (CD34+: subset I) showed a significantly higher level of bcl-2 expression (p <0.0001) together with a lower reactivity for APO 2.7 (p=0.02) as compared to the other more mature CD34- cell subsets. The expression of Bax parallelled that of APO 2.7, although the difference between immature CD34+ blast cells and the mature blast cell subsets did not reach statistical significance (p=0.18). These results translated into a significantly (p<0.0001) higher bcl-2/bax ratio for the CD34+ blast cells as compared to that of the two CD34- blast cell subpopulations. Regarding the expression of the multidrug resistance-associated proteins Pgp and MRP, CD34+ blast cells displayed a greater expression of both proteins as compared to the more mature CD34- AML blast cells, but differences according to maturation stage of AML blast cells did not reach statistical significance. In contrast, LRP expression was significantly lower in the more immature CD34+ blast cell subset than in the more mature ones (p=0.01). INTERPRETATIONS AND CONCLUSIONS: As far as normal bone marrow is concerned our results suggest that all blast cell subpopulations are more protected from apoptosis than their normal counterparts. We conclude that in elderly patients with AML the more immature blast cells are more resistant to apoptotic processes, which could explain why, when AML relapses, the blast cells frequently display a more immature phenotype than that observed at diagnosis. Contradictory results in multidrug resistance profile support the hypothesis that failure to respond to chemotherapeutic drugs in AML is a multifactorial phenomenon. PMID- 11726323 TI - Factor V Leiden mutation and the risk of venous thromboembolism in pregnant women. AB - BACKGROUND AND OBJECTIVES: In this retrospective, single center, cohort study we assessed the risk of pregnancy-related venous thromboembolism (VTE) in women belonging to a large number of families identified because of a symptomatic proband with single identified factor V Leiden mutation. DESIGN AND METHODS: Female family members who had experienced at least one full-term pregnancy were enrolled in the study. Two hundred and seventy pregnancies occurred in 105 carriers and 215 pregnancies in 81 non-carriers of factor V Leiden mutation. RESULTS: The frequency of VTE was 6.4% for heterozygous, 16.7% for homozygous, 20% for double heterozygous carriers of thrombophilic defects, and 1.2% for non carriers. The majority of VTE events related to pregnancy occurred in the post partum period. The relative risks of developing pregnancy-related VTE in women who were carriers of heterozygous and homozygous (or combined heterozygous) factor V Leiden mutation as compared to non-carriers were 5.3 (95% CI, 0.6 to 43.9) and 15.4 (95% CI, 1.4 to 164), respectively. INTERPRETATION AND CONCLUSIONS: Factor V Leiden mutation is a risk factor for pregnancy-related VTE, especially in its homozygous form and in combination with other thrombophilic abnormalities. Screening of families with this mutation might be useful for women of fertile age, as they may take advantage from thromboprophylaxis during pregnancy and the post-partum period. PMID- 11726322 TI - Cell cycle regulatory proteins and apoptosis in B-cell chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: The pathogeny of B-cell chronic lymphocytic leukemia (B-CLL) involves both deregulated proliferation and inhibition of cell death. A particular role in the regulation of these phenomena is played by proteins involved in early G1 phase regulation: pRb kinases: cyclin-dependent kinases (cdk): cdk4 and cdk6 activated by cyclins D, and universal cdk inhibitor p27(Kip1). DESIGN AND METHODS: We determined by flow cytometry the expression of p27(Kip1) and cyclins D (D2 and D3) in populations of peripheral blood lymphocytes obtained from 59 (for p27(Kip1)) and 31 (for cyclins D) previously untreated patients with B-CLL, and compared them with cell cycle parameters, cell viability and apoptosis in 72-hour cultures in medium only. As a control we determined the expression of p27(Kip1), cyclin D2 and D3 in peripheral blood CD5+/CD19+ lymphocytes from 15 healthy donors. RESULTS: p27(Kip1) was present in nearly 100% of lymphocytes in all B-CLL populations tested. Its cellular content estimated semiquantitatively by specific mean fluorescence intensity was higher than in normal CD5+/CD19+ lymphocytes, p27(Kip1) was inversely correlated with patients' age and not correlated with other clinical variables, cell cycle or apoptosis rate. Cyclin D2 was detectable in 25 out of 31, and cyclin D3 in all B CLL lymphocytes populations studied. In contrast to p27Kip1 present in all CD5+/CD19+ lymphocytes, both cyclins were detected only in a subset of neoplastic cells: 27.5 to 87% (mean 51.2) for cyclin D2 and 20.3 to 98% (mean 76.5) for cyclin D3. In cyclin D2- and D3-positive normal CD5+/CD19+ lymphocytes and B-CLL cell populations, cyclin D3 was expressed in a higher percentage of cells than cyclin D2. Both cyclin D2-and cyclin D3-positive fractions of B-CLL cells were, on average, larger than corresponding fractions of normal CD5+/CD19+ peripheral blood lymphocytes. INTERPRETATION AND CONCLUSIONS: Our results indicate that cyclin D3 plays an important role in the regulation of normal and neoplastic CD5+/CD19+ cells, and point to the possibility of the exit of a number of CLL lymphocytes from quiescence. PMID- 11726324 TI - Thalassemia screening based on red cell indices in the Chinese. PMID- 11726326 TI - Application of a chemiluminescent methodology for detection of minimal residual disease in childhood acute lymphoblastic leukemia. PMID- 11726325 TI - Polyploidy in acute promyelocytic leukemia without the 15:17 translocation. PMID- 11726327 TI - High-dose therapy and autologous stem cell transplantation for high-risk Hodgkin's lymphoma: a single center experience. PMID- 11726328 TI - The irreplaceable image: Acute toxicity in erythroid bone marrow progenitors after antimonial therapy. PMID- 11726329 TI - Tge irreplaceable image: Cutaneous mucormycosis in acute myeloid leukemia; hit hard and quickly. PMID- 11726330 TI - The irreplaceable image: Amelioration of oral mucosal lesions of acute graft versus-host disease by low-level laser therapy. PMID- 11726331 TI - The irreplaceable image: Pulmonary graft-versus-host disease. PMID- 11726332 TI - The role of the occupational physician in substance abuse. AB - Occupational physicians are in a unique position to intervene early in substance abuse in the workplace. Despite the complexities of and the barriers to identifying and approaching the troubled worker, there are a number of effective methods available to enable a successful intervention and an appropriate referral to the substance abuse treatment system. Motivational interviewing is one effective way to increase motivation, in a short span of time, in a worker facing needed change. Drug testing in the workplace abounds with potential conflict. Guidelines set forth by the ADA and ACOEM can help the occupational physician balance the need to advocate and protect the rights and confidentiality of patients with the need to uphold the well being and safety of the workplace and of the general public. PMID- 11726333 TI - Substance abuse in the workplace: epidemiology, effects, and industry response. AB - The Substance Abuse and Mental Health Services Administration's National Household Survey on Drug Abuse (NHSDA) reveals self-reported information on illicit drug and alcohol use among full-time, part-time, and unemployed U.S. workers, including information on workplace policies, workers' health, productivity, absenteeism, job turnover rates, accidents, and injuries. Selected statistics from 1985, 1993, and 1999 NHSDAs are reviewed in this chapter, with focus on the effectiveness and outcomes of a comprehensive Drug-Free Workplace Program, including industry and employee response and the effects of a drug testing program. PMID- 11726334 TI - The substance abuse treatment system in the U.S. What is it? What does it do? Myths and misconceptions. AB - The current system of care for treating individuals with substance use disorders evolved during the second half of the 20th century. It has roots in self-help movements and in medical and scientific research findings, and has been shaped by emerging drug trends, public health problems, and uneven treatment financing policies. Currently, the bulk of care is provided by: structured outpatient treatment programs; narcotic treatment programs; variable length residential programs (with and without medical services); and modified therapeutic communities. The majority of the services provided within these organizations meet professional credentialing and/or licensing requirements. The treatment delivery system has made great strides in the past decade to improve quality of care, incorporate outcomes monitoring, and increase the role of empirically supported treatments. Research on the effectiveness of substance abuse treatment indicates that the success rates for these services are comparable to other areas of medical/psychiatric care. PMID- 11726335 TI - Addiction: selecting appropriate treatment and using the self-help system. AB - This chapter provides guidance for the occupational physician in identifying appropriate resources within the treatment and self-help system. Identifying resources to meet patient needs can be challenging in a climate where services are rapidly changing. Two inpatient or outpatient programs may have considerably different services, strengths, and limitations that can be difficult to precisely characterize. Degree of structure (e.g., inpatient versus outpatient) may be misleading in terms of the nature of the treatment provided. Knowledge and attitudes about the self-help system vary greatly, and it is important not to underestimate its potential or hold unrealistic expectations for what it can accomplish. PMID- 11726336 TI - Motivational interviewing in substance abuse: applications for occupational medicine. AB - Motivational interviewing has been proven to increase the motivation of individuals who need to change substance abuse and other health behaviors. This article explores a descriptive model of change and describes how motivational interviewing fits in this larger context. The theoretical assumptions of motivational interviewing are described, including controversies surrounding denial and resistance in ambivalent individuals. Specific and concrete strategies are provided for occupational medicine settings and various combinations of motivational interviewing with feedback and other treatments are described. PMID- 11726337 TI - Substance use disorders: approaching the patient traditional history and physical, or screening? AB - Physicians are encouraged either to assess patients via the traditional history and physical examination for substance abuse issues, or to screen them to determine who is at risk for developing problems related to drug or alcohol use. The former approach is more time consuming and requires some training for the physician to become adept, yet it is ultimately more rewarding both for the patient and physician. Screening and referral is quicker, but also requires some training and an awareness of resources to which the patient at high risk can be referred. This chapter offers basic guidance for both approaches: how to take a productive history, what to look for in a physical examination, which laboratory tests are most helpful; plus standard screening questions and referral indices. PMID- 11726338 TI - Drug testing in the workplace: integrating medical review officer duties into occupational medicine. AB - The authors discuss the social and political climate of drug testing in the workplace, the role of the medical review officer, Federal guidelines, and toxicology methods. The Americans with Disabilities Act is also briefly discussed, and the chapter closes with a case study. PMID- 11726339 TI - Medications in the treatment of addiction: workplace issues. AB - Despite resistance from some treatment providers, medications play an increasingly important role in the treatment of substance dependence disorders. Goals of pharmacotherapy are discussed in terms of a four-stage model of treatment: acute withdrawal, protracted withdrawal, abstinence, and maintenance. The greatest controversy centers on the use of medications during the abstinence and maintenance phases of recovery. Particularly controversial, despite considerable supporting scientific data, is the lon -term use of agonists such as methadone. Occupational physicians and other non-prescribing staff can play a crucial role by supporting the responsible use of medications, maintaining contact with other medical and psychosocial treatment providers, enlisting support for medication compliance in patients' personal and professional support systems, advocating for comprehensive and humane patient care, and being alert to signs of medication misuse. PMID- 11726340 TI - A model comprehensive psychiatric fitness-for-duty evaluation. AB - A three-pronged model for a comprehensive psychiatric fitness-for-duty evaluation is presented as a resource tool for occupational physicians in the evaluation of substance abuse. Specialized psychiatric evaluators can play a role in (1) rendering diagnosis and fitness-for-duty determinations, (2) recommending treatment and monitoring plans, and (3) offering remedies for improving the employee/employer and medical systems. The comprehensive evaluation is especially helpful in difficult-to-diagnose and complex cases. The author provides guidance for deciding when to refer, discusses issues of confidentiality, and lists indicators of substance abuse based on employee behavior/performance and medical signs and symptoms. PMID- 11726341 TI - Legal issues facing physicians: substance abuse in the workplace. AB - This chapter addresses key Department of Transportation (DOT) rules for drug testing employees working in industries regulated by the DOT. It also discusses the issue of substance abuse and the Americans with Disabilities Act. Finally, it reviews the iss e of confidentiality. It is important for the reader to recognize that this chapter is but a review of critical and crucial material. The reader is encouraged to pursue additional information if greater clarity and understanding are required. PMID- 11726342 TI - Tobacco in the workplace. AB - Tobacco use, the leading cause of preventable death, has numerous adverse impacts in the workplace. It is associated with increased healthcare costs and disability, greater absenteeism, decrements in job performance, and increased risk of injury, and secondhand smoke is a major pollution problem. Tobacco dependence responds to treatment, and employers have several opportunities to offer treatment to employees and their families. Smoke-free environments can help smokers cut back or quit. Combining sound workplace policies and treatment with community involvement in tobacco control provides synergies to reduce tobacco use in the community as well as in the workforce. These activities benefit workers and their families, employers, and society at large. PMID- 11726344 TI - Management of Fever in neutropenic patients with cancer. AB - During the last decade, significant advancements have been made in supportive care of neutropenic patients with cancer, including broad-spectrum antimicrobial agents and hematopoietic growth factors. Despite these achievements, infection continues to be the major cause of morbidity and mortality in patients with cancer because of alterations of normal host defenses. Because infection in patients with immunocompromised cancer is a potentially life-threatening situation, it is imperative that clinicians have optimal knowledge of therapeutic interventions directed toward the prevention, detection, and treatment of infection. Furthermore, the clinician should have basic knowledge about the pathophysiology of febrile neutropenia. PMID- 11726345 TI - Deep vein thrombosis and anticoagulant therapy. AB - This article focuses on deep vein thrombosis (DVT) and anticoagulant therapy. The risk factors, pathophysiology, prevention, and detection of DVT are explained. Anticoagulants such as unfractionated heparin (UFH), low molecular weight heparin (enoxaparin), and warfarin are discussed. The article also reviews nursing assessment and monitoring of patients with DVT as well as patient education on anticoagulants. PMID- 11726346 TI - Management of malignant pleural effusion. AB - Malignant pleural effusion, an abnormal accumulation of fluid in the intrapleural space, is a common complication of cancer resulting in debilitating symptoms including dyspnea, pain, and fatigue. The goal of treatment is aimed at symptomatic control, relief and control of dyspnea, and obliteration of the intrapleural space to prevent reaccumulation of pleural fluid. Treatment options include chemotherapy, radiation, intrapleural instillation of sclerosing agents, indwelling pleural catheters with intermittent drainage, pleuroperitoneal shunts, and pleurectomy. Therapeutic approaches used in the management of malignant effusions must be directed at maintaining or improving the quality of life for the individual. PMID- 11726347 TI - Fatigue: a debilitating symptom. AB - Fatigue is a commonly reported and observed symptom among patients at the University of Texas M.D. Anderson Cancer Center. Caring for patients with this debilitating symptom is a major challenge. The nurse practitioner in the Fatigue Clinic has a special role in the management of these patients. PMID- 11726348 TI - Management of chemotherapy-induced diarrhea and constipation. AB - This article reviews the incidence, etiology, pathophysiology, and assessment of diarrhea and constipation related to chemotherapy. Protocols for treatment are presented. The nurse's role in prevention, patient education, assessment, and treatment is emphasized. PMID- 11726349 TI - Lymphedema management in patients with lymphoma. AB - Lymphoma and its treatments put patients at risk for lymphedema. The effect of lymphedema and its treatments on quality of life has been studied in other populations, but not in patients with lymphoma. Treatment effectiveness is poorly documented and confusion surrounding lymphedema therapy creates barriers to care. Reimbursement issues and the paucity of therapists limit access to care. Nursing research, care and collaboration with lymphedema therapists directly impacts the quality of lymphedema care for patients with lymphoma. Lymphedema pathology and treatments are described and recommendations are made for nursing care and research. PMID- 11726350 TI - Anorexia and cachexia in advanced cancer. AB - During the last days of life, patients experience a myriad of symptomatology. While the exact percentage of patients with cancer affected by anorexia may be subject to debate, there is a clear indication that a significant portion of patients with cancer will at some point in the course of his or her illness suffer the ravages of anorexia and the related progressive weight loss that accompanies it. Anorexia is often related to the tissue wasting process of cachexia, a more severely debilitating condition that may be a contributing factor, or even the primary cause of death, in approximately 20% of cancer patients. Clinicians involved in oncology and hospice/palliative care should have a clear understanding of this process and appropriate interventions for patients experiencing anorexia/cachexia. PMID- 11726351 TI - Mucositis in the oncology patient. AB - Mucositis is a common side effect related to the treatment of cancer. Many agents have been tested for the prevention and management of mucositis, but the data regarding a number of the agents are conflicting. This article describes the incidence of mucositis in individuals with cancer and the various agents used in its prevention and management. Nurses play an important role in the management of patients with mucositis and in identifying agents that may decrease the patient's risk for mucositis and aid healing. PMID- 11726352 TI - Palliation of gastrointestinal obstructive disorders. AB - Gastrointestinal and biliary obstructive disorders are complications encountered in advanced cancer nursing that are associated with significant morbidity and mortality. Patients with bowel gastric, or hepatobiliary obstruction require prompt and accurate diagnosis, so that appropriate care is initiated to treat the obstruction and related symptoms. Intensive nursing care aimed at comfort, psychological support, and patient/family education is essential for the well being of patients suffering from a gastrointestinal obstructive disorder. PMID- 11726353 TI - Managing pain at the end of life. AB - There are many threats to a calm and peaceful passing, but none more distressing than unrelenting pain. Pain exists not only in the physical realm, but also in the psychological, social and spiritual senses. Discussion of barriers and responsibilities are important to assure that pain is appropriately treated. Effective pain treatment relies on communication among patients, families, and care providers. Nurses are a central force in this communication and must act as leaders and advocates in planning care and relieving pain in dying patients. PMID- 11726354 TI - Family coping, styles, and challenges. AB - Creating a therapeutic partnership with patients and families during their crisis with cancer is both a challenging and extremely rewarding experience for the oncology nurse. Each family brings to the experience its own structure, maturational stage, culture, and history. They bring individual and group emotional responses, coping strategies and conflicts when confronting a disease over which they have no control. This article describes the nature of contemporary families within the context of initial and ongoing nursing assessment. It also provides examples of family cancer experiences and the therapeutic approaches selected by nurses faced with the challenges of individual and family coping styles. PMID- 11726355 TI - The psychologic distress in patients with cancer. AB - The psychologic distress in patients with cancer may be divided into three categories; depression, anxiety and neuro-cognitive changes including delirium. The supportive interventions of health care professionals along with the use of psychotropic medications have been shown to relieve depression, anxiety, and states of confusion in oncology patients. Oncology nurses are in a critical position to identify problems and intervene to provide appropriate symptom relief. The management of psychologic distress may improve adherence to treatment programs, reduce toxicity, achieve a better quality of life, and enhance grace and dignity at the end of life. PMID- 11726356 TI - Ethical issues confronting oncology nurses. AB - The focus of this article is the ethical dilemmas challenging oncology nurses and their patients. The cancer diagnosis itself may present ethical issues related to genetic testing, prevention and early detection, and information sharing. Cancer treatment options may involve considerations related to level of appropriate care, informed consent, patient education, and unproven, but promising, therapies. Finally, the ethical issues related to outcomes of the disease and treatment, such as pain control, advanced directives, do not resuscitate order, discontinuation of medically inappropriate care, euthanasia, and physician assisted suicide are discussed. PMID- 11726357 TI - Barriers to palliative and supportive care. AB - The goal of palliative care is to provide comprehensive and intense symptom management and psychological, educational, social, and spiritual support. Barriers to palliative care that nurses encounter may be conceptualized into three main levels: the patient/family, professional, and system barriers. Recommendations for nurses, health care policy makers, and other health professionals to address the barriers are discussed. Future research and work needs to be implemented at all three levels to optimize the quality end-of-life care. PMID- 11726358 TI - Self-care for the caregiver. AB - Palliative care nurses can face unique stressors and compassion fatigue working in their field. Working with the dying and their families, communicating with other health care professionals, and handling ethical issues are often sources of stress in palliative care. The biochemistry and theory of stress are discussed. Various strategies to cope with caregiver stress are explained. PMID- 11726359 TI - Biopreparedness and public health. PMID- 11726360 TI - Land use history and public health. PMID- 11726362 TI - The interpretation of statistically significant results: the impact of the Taiwan helmet use law. PMID- 11726363 TI - Individualized or population risks: what is the argument? PMID- 11726366 TI - The challenge and place of international human rights in public health. PMID- 11726367 TI - Can public health researchers and agencies reconcile the push from funding bodies and the pull from communities? AB - Responding to growing impatience with the limited application of research findings to health practices and policies, both funding bodies and communities are demanding that research show greater sensitivity to communities' perceptions, needs, and unique circumstances. One way to assure this is to employ participatory research-to engage communities at least in formulating research questions and interpreting and applying research findings and possibly also in selecting methods and analyzing data. "Community" should be interpreted broadly as all who will be affected by the research results, including lay residents of a local area, practitioners, service agencies, and policymakers. Participatory research should not be required of every project, but when results are to be used for, in, and by communities, those communities should collaborate not only in applying findings but also in determining the ways in which the findings are produced and interpreted. PMID- 11726368 TI - What is community? An evidence-based definition for participatory public health. AB - Increased emphasis on community collaboration indicates the need for consensus regarding the definition of community within public health. This study examined whether members of diverse US communities described community in similar ways. To identify strategies to support community collaboration in HIV vaccine trials, qualitative interviews were conducted with 25 African Americans in Durham, NC; 26 gay men in San Francisco, Calif; 25 injection drug users in Philadelphia, Pa; and 42 HIV vaccine researchers across the United States. Verbatim responses to the question "What does the word community mean to you?" were analyzed. Cluster analysis was used to identify similarities in the way community was described. A common definition of community emerged as a group of people with diverse characteristics who are linked by social ties, share common perspectives, and engage in joint action in geographical locations or settings. The participants differed in the emphasis they placed on particular elements of the definition. Community was defined similarly but experienced differently by people with diverse backgrounds. These results parallel similar social science findings and confirm the viability of a common definition for participatory public health. PMID- 11726369 TI - The role of community advisory boards: involving communities in the informed consent process. AB - Ethical research involving human subjects mandates that individual informed consent be obtained from research participants or from surrogates when participants are not able to consent for themselves. The existing requirements for informed consent assume that all study participants have personal autonomy; fully comprehend the purpose, risks, and benefits of the research; and volunteer for projects that disclose all relevant information. Yet contemporary examples of lapses in the individual informed consent process have been reported. The authors propose the use of community advisory boards, which can facilitate research by providing advice about the informed consent process and the design and implementation of research protocols. These activities could help reduce the number of individual informed consent lapses, benefiting study participants and the scientific integrity of the research in question. PMID- 11726370 TI - Urban history, urban health. AB - Over the course of the 20th century, the United States became an urban nation: 80% of Americans now live in metropolitan areas. Supplying basic sanitary services-drinking water, sewers, and garbage removal-to these cities is a gargantuan task, yet most people have little understanding of urban infrastructure systems and their enormous regional ecologic impacts. Municipalization of sanitary services, especially since 1880, distanced people from their wastes and gave city dwellers a simplistic experience of one-way material flow through cities, without knowledge of the environmental costs. Most sanitary infrastructures were built primarily for durability and lack the elasticity to meet changing needs. The challenge now is to adapt sanitary systems for flexibility and simultaneously move from unchecked material consumption toward resource-based thinking. PMID- 11726371 TI - The face of public health nursing: Carol Maureen Easley Allen. PMID- 11726372 TI - Meeting the data needs of a local health department: the Los Angeles County Health Survey. AB - Monitoring the health status of populations is a core function of all public health agencies but is particularly important at the municipal and community levels, where population health data increasingly are used to drive public health decision making and community health improvement efforts. Unfortunately, most local health jurisdictions lack important data for developing population health profiles, such as data on chronic disease prevalence, quality-of-life measures, functional status, and self-perceptions of health status. In addition, data on important determinants of health, including health behaviors and access to health care services, are rarely available locally. These data frequently are collected in national and state surveys (e.g., the National Health Interview Survey and the Behavioral Risk Factor Surveillance System) and provide critical information to assess progress toward achieving state and national health objectives. The surveys rarely serve local data needs, however, because of insufficient sample size and lack of flexibility to address local health issues. To address gaps in local health data, in 1997 the Los Angeles County Department of Health Services inaugurated the Los Angeles County Health Survey. PMID- 11726373 TI - The influenza pandemic of 1918. PMID- 11726374 TI - Understanding and responding to youth substance use: the contribution of a health and human rights framework. AB - This article examines the utility of a health and human rights framework for conceptualizing and responding to the causes and consequences of substance use among young people. It provides operational definitions of "youth" and "substances," a review of current international and national efforts to address substance use among youths, and an introduction to human rights and the intersection between health and human rights. A methodology for modeling vulnerability in relation to harmful substance use is introduced and contemporary international and national responses are discussed. When governments uphold their obligations to respect, protect, and fulfill human rights, vulnerability to harmful substance use and its consequences can be reduced. PMID- 11726375 TI - A bold new direction for environmental health research. AB - The biotechnology revolution has opened new opportunities for addressing current inadequacies in decision making regarding environmental health. Strategic investments need to be made (1) to develop high-throughput technologies that could accelerate toxicity testing and generate a mechanistic understanding of toxicity, (2) to incorporate individual susceptibility into risk assessments, and (3) to establish a rational basis for testing and regulatory decision making. New initiatives of the National Institute of Environmental Health Sciences, including the Environmental Genome Project and the Toxicogenomics Center, are discussed. PMID- 11726376 TI - Implications for tobacco control of the multistate tobacco settlement. AB - The 1998 master settlement agreement between major tobacco manufacturers and the US states will have a profound effect on many tobacco industry practices and will significantly influence future settlements with the tobacco industry. This article analyzes the settlement's key provisions pertaining to youth sales, advertising, marketing, and lobbying. It also examines the ways in which the settlement restricts industry practices as well as the many industry practices that remain unregulated. PMID- 11726377 TI - Community advocates in public housing. PMID- 11726378 TI - Blood lead levels in relation to paint and dust lead levels: the lead-safe cambridge program. PMID- 11726379 TI - The impact on clients of a community-based infant mortality reduction program: the National Healthy Start Program Survey of Postpartum Women. AB - OBJECTIVES: This study assessed the effect of the national Healthy Start Program on its clients. METHODS: We used a cross-sectional survey of a sample from Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) rosters of women less than 6 months postpartum who were residents of Healthy Start Program areas. RESULTS: Healthy Start clients revealed higher sociodemographic risk, but not behavioral risk, for adverse pregnancy outcome than other area residents. They did not differ from other residents in receipt of services except for a greater likelihood of receiving case management, using birth control at the time of the interview, and rating their prenatal care more highly. CONCLUSIONS: The Healthy Start Program succeeded in enrolling women at high risk. It had little effect on the immediately concluded pregnancy, but it might influence future outcomes. PMID- 11726380 TI - Grassroots participation, peer education, and HIV prevention by sex workers in South Africa. AB - OBJECTIVES: This microqualitative case study of a community-based peer education program led by sex workers at a South African mine examined the role of grassroots participation in sexual health promotion. METHODS: The study involved in-depth interviews with 30 members of the target community. The interviews were analyzed in terms of social capital, empowerment, and identity. RESULTS: The study yielded a detailed analysis of the way in which community dynamics have shaped the peer education program's development in a deprived, violent community where existing norms and networks are inconsistent with ideal criteria for participatory health promotion. CONCLUSIONS: Much remains to be learned about the complexities of translating theoretically and politically vital notions of "community participation" into practice among hard-to-reach groups. The fabric of local community life is shaped by nonlocal structural conditions of poverty and sexual inequality in ways that challenge those seeking to theorize the role of social capital in community development in general and in sexual health promotion in particular. PMID- 11726381 TI - The visibility of illicit drugs: implications for community-based drug control strategies. AB - OBJECTIVES: This study examined differences between the visibility of drugs and drug use in more than 2100 neighborhoods, challenging an assumption about drug use in poor, minority, and urban communities. METHODS: A telephone survey assessed substance use and attitudes across 41 communities in an evaluation of a national community-based demand reduction program. Three waves of data were collected from more than 42 000 respondents. RESULTS: Measures of neighborhood disadvantage, population density, and proportion of minority residents explained more than 57% of the variance between census tracts in visibility of drug sales but less than 10% of tract-to-tract variance in drug use. Visible drug sales were 6.3 times more likely to be reported in the most disadvantaged neighborhoods than in the least disadvantaged, while illicit drug use was only 1.3 times more likely. CONCLUSIONS: The most disadvantaged neighborhoods have the most visible drug problems, but drug use is nearly equally distributed across all communities. Thus, efforts to address drug-related problems in poorer areas need to take into account the broader drug market served by these neighborhoods. PMID- 11726382 TI - Environmental and policy determinants of physical activity in the United States. AB - OBJECTIVES: This study examined (1) descriptive patterns in perceived environmental and policy determinants of physical activity and (2) associations between these factors and behavior. METHODS: A cross-sectional study was conducted from 1999 to 2000 among US adults; individuals at lower income levels were oversampled. RESULTS: Availability of areas for physical activity was generally higher among men than among women. The 4 most commonly reported personal barriers were lack of time, feeling too tired, obtaining enough exercise at one's job, and no motivation to exercise. Neighborhood characteristics, including the presence of sidewalks, enjoyable scenery, heavy traffic, and hills, were positively associated with physical activity. There was a high level of support for health policy-related measures. Up to one third of individuals who had used environmental supports reported an increase in physical activity. CONCLUSIONS: An array of environmental and policy determinants, particularly those related to the physical environment, are associated with physical activity and should be taken into account in the design of interventions. PMID- 11726383 TI - Using signs, artwork, and music to promote stair use in a public building. AB - OBJECTIVES: This study assessed the impact on stair use of improving the attractiveness of a stairwell. METHODS: Observations of stair usage were made in a university building during baseline, 2 interventions, and follow-up. The first intervention involved signs; the second intervention added artwork and music in the stairwell. RESULTS: More participants used the stairs during the music and artwork intervention than at baseline or when signs alone were used. CONCLUSIONS: Improving the aesthetic qualities of a stairwell can increase rates of stair usage in a public building. Designs for buildings should take accessibility and aesthetic issues into consideration. PMID- 11726384 TI - Promoting stair use in a US-Mexico border community. AB - OBJECTIVES: This study sought to determine whether a culturally relevant health message would promote stair use in a predominantly Hispanic community. METHODS: Observations of stair, elevator, and escalator use were collected over a 6-month period at 4 sites throughout the city of El Paso, Tex. The efficacy of individual and family health promotion signs was tested. RESULTS: Stair use increased in response to both individual and family promotion health messages, and use varied widely by intervention site. CONCLUSIONS: These results underscore the importance of considering the physical characteristics of the environments targeted for health promotion campaigns. PMID- 11726385 TI - Limitations on the use of a single screening question to measure sedentary behavior. AB - OBJECTIVES: This study explored the limitations of identifying sedentary individuals via an existing screening question in a state-based surveillance system. METHODS: A national sample (n = 7529) of adults, selected by random-digit dialing between November 1999 and May 2000, responded about participation in leisure-time physical activity. RESULTS: Of those who initially reported no leisure-time physical activity (25%), 85% were engaging in at least some activity, and 20% were engaging in enough moderate- or vigorous-intensity activity to meet health-related recommendations. CONCLUSIONS: Public health programs that use only 1 screening question to identify sedentary behavior may not be able to target physical activity messages effectively, especially if physical activity is defined to include a broad range of activities beyond sports. PMID- 11726386 TI - A population-based assessment of human rights abuses committed against ethnic Albanian refugees from Kosovo. AB - OBJECTIVES: This study assessed patterns of displacement and human rights abuses among Kosovar refugees in Macedonia and Albania. METHODS: Between April 19 and May 3, 1999, 1180 ethnic Albanian refugees living in 31 refugee camps and collective centers in Macedonia and Albania were interviewed. RESULTS: The majority (68%) of participants reported that their families were directly expelled from their homes by Serb forces. Overall, 50% of participants saw Serb police or soldiers burning the houses of others, 16% saw Serb police or soldiers burn their own home, and 14% witnessed Serb police or soldiers killing someone. Large percentages of participants saw destroyed mosques, schools, or medical facilities. Thirty-one percent of respondents reported human rights abuses committed against their household members, including beatings, killings, torture, forced separation and disappearances, gunshot wounds, and sexual assault. CONCLUSIONS: The present findings confirm that Serb forces engaged in a systematic and brutal campaign to forcibly expel the ethnic Albanian population of Kosovo. In the course of these mass deportations, Serb forces committed widespread abuses of human rights against ethnic Albanians. PMID- 11726387 TI - The relation between community bans of self-service tobacco displays and store environment and between tobacco accessibility and merchant incentives. AB - OBJECTIVES: These studies investigated (1) the effect of community bans of self service tobacco displays on store environment and (2) the effect of consumer tobacco accessibility on merchants. METHODS: We counted cigarette displays (self service, clerk-assisted, clear acrylic case) in 586 California stores. Merchant interviews (N = 198) identified consumer tobacco accessibility, tobacco company incentives, and shoplifting. RESULTS: Stores in communities with self-service tobacco display bans had fewer self-service displays and more acrylic displays but an equal total number of displays. The merchants who limited consumer tobacco accessibility received fewer incentives and reported lower shoplifting losses. In contrast, consumer access to tobacco was unrelated to the amount of monetary incentives. CONCLUSIONS: Community bans decreased self-service tobacco displays; however, exposure to tobacco advertising in acrylic displays remained high. Reducing consumer tobacco accessibility may reduce shoplifting. PMID- 11726388 TI - Reactive oxygen species and cell signaling. PMID- 11726389 TI - AP endonucleases and the many functions of Ref-1. PMID- 11726390 TI - Surfactant protein A (SP-A)-mediated bacterial clearance: SP-A and cystic fibrosis. PMID- 11726391 TI - A novel LPS-inducible CCR3 activator: why so many CCR3 ligands? PMID- 11726392 TI - Ceramide-mediated apoptosis in lung epithelial cells is regulated by glutathione. AB - Reactive oxygen species (ROS) are mediators of lung injury, and glutathione (GSH) is the major nonprotein antioxidant that protects the cell from oxidative stress. We have recently shown that H(2)O(2) induces ceramide-mediated apoptosis in human lung epithelial cells. We hypothesized that ROS-mediated depletion of GSH plays a regulatory role in ceramide generation, and thus in the induction of apoptosis. Our present studies demonstrate that GSH at physiologic concentrations (1 to 10 mM) inhibits ceramide production in a time- and dose-dependent manner in A549 human alveolar epithelial cells. On the other hand, buthionine-sulfoximine mediated depletion of intracellular GSH induces elevation of ceramide levels and apoptosis. In addition, GSH blocks H(2)O(2)-mediated induction of intracellular ceramide generation and apoptosis. These effects were not mimicked by oxidized GSH (GSSG) or other thiol antioxidants, such as dithiothreitol and 2 mercaptoethanol. Moreover, increase of intracellular H(2)O(2), mediated by inhibition of catalase by aminotriazole, also induces ceramide generation and apoptosis. These effects were blocked by N-acetylcysteine. Our results suggest that GSH depletion may be the link between oxidative stress and ceramide-mediated apoptosis in the lung. PMID- 11726393 TI - Secretion of interleukin-1 receptor antagonist from human mast cells after immunoglobulin E-mediated activation and after segmental antigen challenge. AB - Mast cells produce substances with antiinflammatory properties in addition to their capacity to release proinflammatory mediators. To further probe the antiinflammatory aspect of mast-cell function we investigated the ability of human mast cells (huMCs) to produce interleukin (IL)-1 receptor antagonist (IL 1ra) in response to high-affinity Fc receptor for immunoglobulin E (Fcalpha RI) aggregation, and examined IL-1ra in bronchoalveolar lavage fluid (BALF) to determine whether it might be of mast-cell origin. Using a ribonuclease protection assay, flow cytometry, and enzyme-linked immunosorbent assay (ELISA), IL-1ra message and protein were found to be constitutively expressed in cultured huMCs. Upon stimulation through Fcalpha RI, IL-1ra message was upregulated in huMCs and IL-1ra protein secreted from cultured huMCs and isolated human lung mast cells. By immunoblot analysis, huMCs were found to produce the 17-kD form of IL-1ra and the presence of IL-1ra in human lung mast cells was confirmed by immunohistochemistry. In BALF obtained from allergic asthmatic subjects, IL-1ra production increased after specific antigen challenge, with the 17-kD isoform of IL-1ra predominating. These findings demonstrate that huMCs produce and release IL-1ra after Fcalpha RI aggregation, which may contribute to a local inhibition of IL-1-dependent effects on inflammation in the lung. PMID- 11726394 TI - Expression of yeast apurinic/apyrimidinic endonuclease (APN1) protects lung epithelial cells from bleomycin toxicity. AB - Bleomycin is a well-established anti-tumor drug. Its major untoward effect, pulmonary toxicity, has limited its usage. In this study, we used a DNA repair protein, yeast apurinic/apyrimidinic endonuclease (APN1) to reduce the toxicity of bleomycin on lung cells. A549 cells, an alveolar epithelial cell line, were transduced by MIEG3 retroviral vector encoding both enhanced green fluorescent protein (EGFP) and APN1. Transduced cells were sorted by fluorescent-activated cell sorter (FACS) analysis and were cloned. The APN1 expression of transduced A549 cell population and four selected clones expressing different levels of EGFP was confirmed by Northern, Western, and apurinic/apyrimidinic (AP) endonuclease activity analyses. The expression of APN1 was positively correlated with the expression of EGFP. The protective effect of APN1 against bleomycin was determined by single cell gel electrophoresis/Comet assay and by clonogenic survival assay following bleomycin treatment. The A549 population expressing APN1 showed a significant reduction of DNA damage in the presence of 20, 50, and 100 microg/ml bleomycin; similarly, the APN1-expressing A549 population also demonstrated increased survival in the presence of bleomycin compared with the vector-transduced A549 population. In selected clones, three of four APN1 expressing clones resulted in significantly improved cell survival. The current study suggests that the yeast DNA repair protein, APN1, can reduce bleomycin toxicity to target lung cells. PMID- 11726395 TI - Effects of surfactant protein A and NaCl concentration on the uptake of Pseudomonas aeruginosa by THP-1 cells. AB - In the present study, we characterized a model system in which we examined the effects of human surfactant protein A (SP-A) on the uptake of a common human pulmonary pathogen, Pseudomonas aeruginosa, by a human monocytic/macrophage cell line, THP-1 cells. We found that SP-A significantly increases uptake of the bacteria in a dose-dependent manner. Bacterial uptake was temperature-dependent, because an effect of SP-A on bacterial uptake was observed at 37 degrees C and not at 4 degrees C. The continued presence of SP-A during the period when the bacteria and THP-1 cells were co-incubated was necessary for enhanced uptake. Pre incubation of the bacteria or THP-1 cells with SP-A, followed by washing, abolished the effect of SP-A on bacterial uptake. The effect of SP-A could be inhibited by high concentrations of mannose, but was not affected by the removal or addition of lipopolysaccharide (LPS). Finally, we observed that the SP-A mediated increase in uptake of P. aeruginosa by THP-1 cells was optimal in a narrow (100 mM and 150 mM) range of NaCl concentrations. We conclude that SP-A enhances the THP-1 cell-mediated uptake of P. aeruginosa in a manner dependent on temperature, the concentration of SP-A, and the concentration of NaCl. PMID- 11726396 TI - LPS induces eosinophil migration via CCR3 signaling through a mechanism independent of RANTES and Eotaxin. AB - Mounting evidence suggests that lipopolysaccharide (LPS) modulates bronchoconstriction and eosinophil function in asthma. We have investigated the role of different chemokines in the eosinophil influx to the pleural cavity after LPS stimulation. Expression of mRNA for eotaxin, regulated on activation, normal T cells expressed and secreted (RANTES), macrophage inflammatory protein (MIP) 1alpha, MIP-1beta, MIP-2, and monocyte chemotactic protein (MCP)-1 was increased in cells recovered from the mouse pleural cavity 6 h after LPS administration. Eotaxin and RANTES, but not MIP-1alpha, protein levels were also increased in cell-free pleural washes recovered 6 h after LPS stimulation (LPW). Antimurine eotaxin and antimurine RANTES antibodies (Abs) failed to inhibit LPS-induced eosinophil influx into mouse pleural cavity in vivo. Pertussis toxin inhibited LPW-induced eosinophil shape change in vitro, suggesting the involvement of G protein-coupled receptors in LPW signaling. Blockade of CCR3 receptors diminished eosinophil shape change induced by LPW fractions in vitro and LPS-induced eosinophil accumulation in vivo. To investigate further contribution of CC chemokines, we administered a 35-kD CC chemokine neutralizing protein (vCKBP) in vivo. vCKBP inhibited the eosinophil accumulation induced by eotaxin and ovalbumin, but did not block that induced by LPS or LPW. Our data suggest that LPS-induced eosinophil accumulation depends on G protein-coupled CCR3 receptor activation, through a mechanism independent of eotaxin, RANTES, or other vCKBP inhibitable CC chemokines. PMID- 11726397 TI - Ventilator-induced lung injury upregulates and activates gelatinases and EMMPRIN: attenuation by the synthetic matrix metalloproteinase inhibitor, Prinomastat (AG3340). AB - Mechanical ventilation has become an indispensable therapeutic modality for patients with respiratory failure. However, a serious potential complication of MV is the newly recognized ventilator-induced acute lung injury. There is strong evidence suggesting that matrix metalloproteinases (MMPs) play an important role in the development of acute lung injury. Another factor to be considered is extracellular matrix metalloproteinase inducer (EMMPRIN). EMMPRIN is responsible for inducing fibroblasts to produce/secrete MMPs. In this report we sought to determine: (1) the role played by MMPs and EMMPRIN in the development of ventilator-induced lung injury (VILI) in an in vivo rat model of high volume ventilation; and (2) whether the synthetic MMP inhibitor Prinomastat (AG3340) could prevent this type of lung injury. We have demonstrated that high volume ventilation caused acute lung injury. This was accompanied by an upregulation of gelatinase A, gelatinase B, MT1-MMP, and EMMPRIN mRNA demonstrated by in situ hybridization. Pretreatment with the MMP inhibitor Prinomastat attenuated the lung injury caused by high volume ventilation. Our results suggest that MMPs play an important role in the development of VILI in rat lungs and that the MMP inhibitor Prinomastat is effective in attenuating this type of lung injury. PMID- 11726398 TI - Phosphatidylcholine molecular species in lung surfactant: composition in relation to respiratory rate and lung development. AB - Surfactant reduces surface tension at the air-liquid interface of lung alveoli. While dipalmitoylphosphatidylcholine (PC16:0/ 16:0) is its main component, proteins and other phospholipids contribute to the dynamic properties and homeostasis of alveolar surfactant. Among these components are significant amounts of palmitoylmyristoylphosphatidylcholine (PC16:0/ 14:0) and palmitoylpalmitoleoylphosphatidylcholine (PC16:0/ 16:1), whereas in surfactant from the rigid tubular bird lung, PC16:0/14:0 is absent and PC16:0/16:1 strongly diminished. We therefore hypothesized that the concentrations of PC16:0/14:0 and PC16:0/16:1 in surfactants correlate with differences in the respiratory physiology of mammalian species. In surfactants from newborn and adult mice, rats, and pigs, molar fractions of PC16:0/14:0 and PC16:0/16:1 correlated with respiratory rate. Labeling experiments with [methyl-(3)H]choline in mice and perfused rat lungs demonstrated identical alveolar proportions of total and newly synthesized PC16:0/14:0, PC16:0/16:1, and PC16:0/16:0, which were much higher than those of other phosphatidylcholine species. In surfactant from human term and preterm neonates, fractional concentrations not only of PC16:0/16:0 but also of PC16:0/14:0 and PC16:0/ 16:1 increased with maturation. Our data emphasize that PC16:0/14:0 and PC16:0/16:1 may be important surfactant components in alveolar lungs, and that their concentrations are adapted to respiratory physiology. PMID- 11726399 TI - Reduced Smad3 protein expression and altered transforming growth factor-beta1 mediated signaling in cystic fibrosis epithelial cells. AB - Cystic fibrosis (CF) is a disease characterized by an aggressive inflammatory response in the airways. Given the antiinflammatory properties of transforming growth factor (TGF)-beta1, it was our goal to examine components of TGF-beta1 mediated signaling in both a cultured cell model and a mouse model of CF. A CF related reduction of protein levels of the TGF-beta1 signaling molecule Smad3 was found in both of these model systems, whereas Smad4 levels were unchanged. Functional effects of reduced Smad3 expression are manifest in our cultured cell model, as reduced basal and TGF-beta1-stimulated levels of luciferase expression using the TGF-beta1-responsive reporter construct 3TP-Lux in the CF-phenotype cells compared with control cells. However, TGF-beta1-stimulated responses using the A3-Luc reporter construct were normal in both cell lines. These results suggest that select TGF-beta1-mediated signaling pathways are impaired in CF epithelial cells. This selective loss of Smad3 protein expression in CF epithelium may also influence inflammatory responses. Our data demonstrate that both CF-phenotype cells lacking Smad3 expression, and A549 cells expressing a dominant-negative Smad3, are unable to support TGF-beta1-mediated inhibition of either the interleukin (IL)-8 or the NOS2 promoter. We conclude that a CF-related reduction in Smad3 protein expression selectively alters TGF- beta1-mediated signaling in CF epithelium, potentially contributing to aggressive inflammatory responses. PMID- 11726400 TI - Interleukin-13 induces proliferation of human airway epithelial cells in vitro via a mechanism mediated by transforming growth factor-alpha. AB - Remodeling of the airways, as occurs in asthmatic patients, is associated with the continual presence of inflammatory mediators and Th2 cytokines, especially interleukin (IL)-13, during cycles of epithelial injury and repair. In this study, we examined the effect of IL-13 on well-differentiated normal human bronchial epithelial (NHBE) cells maintained in air-liquid interface culture. IL 13 induced proliferation of NHBE cells after 24 h exposure, as reflected by [(3)H]thymidine uptake and cell counts. The effects of IL-13 were mediated through the epidermal growth factor receptor (EGFR), as proliferation was attenuated by AG1478, an EGFR tyrosine kinase inhibitor. Proliferation appeared to be mediated by transforming growth factor (TGF)-alpha, a potent ligand for EGFR, which was released rapidly from NHBE cells in response to IL-13. Neutralizing antibody to TGF-alpha, but not antibodies against other potentially important growth factors (EGF, heparin binding epidermal growth factor-like growth factor [HB-EGF], platelet-derived growth factor [PDGF]), inhibited the mitogenic response to IL-13. This study provides the first experimental evidence that IL-13 can initiate a proliferative response of human airway epithelium in the absence of inflammatory cells or other cell types. The results are consistent with a mechanism whereby IL-13 induces release of TGF-alpha from the epithelial cells, which in turn binds via an autocrine/paracrine-type action to the EGFR, initiating proliferation. IL-13-induced airway remodeling in vivo may involve this epithelium-driven response. PMID- 11726401 TI - Effects of retinoic acid receptor-selective agonists on human nasal epithelial cell differentiation. AB - Retinoids play a critical role in the maintenance of the mucociliary phenotype of epithelial cells in the upper respiratory tract. To determine the role of retinoic acid receptors (RARs) in the regulation of epithelial differentiation, we tested the effect of the synthetic retinoids CD336, CD2019, and CD666, selective agonists for RARalpha, RARbeta, and RARgamma, respectively, during differentiation of human nasal epithelial (HNE) cells in vitro. Using glutamylated tubulin and transglutaminase I (Tg I) as markers of ciliated cell and squamous cell differentiation, respectively, we showed that retinoic acid (RA) stimulated mucociliary differentiation and, in parallel, inhibited squamous cell differentiation. The agonists of the three RARs independently induced ciliogenesis and inhibited squamous cell differentiation by downregulating Tg I expression in a dose- and time-dependent manner. Antagonists specific for the three RARs abolished the effects of the corresponding agonists, demonstrating an RAR-specific mediated effect. Moreover, treatment of retinoid-deficient cultures with RAR agonists induced conversion of the squamous-like phenotype into a ciliated phenotype. In conclusion, all three RARs are potentially involved in the differentiating effects of RA in respiratory epithelial cells. PMID- 11726402 TI - CTLA4-IgG reverses asthma manifestations in a mild but not in a more "severe" ongoing murine model. AB - We investigated whether CTLA4-Ig can reverse established asthma manifestations in a novel murine model of ongoing disease. In BALB/c mice, sensitized to ovalbumin (OVA) without adjuvant, airway inflammation was induced by a first series of OVA aerosol challenges. Murine CTLA4-IgG was then administered, followed by a second series of OVA inhalations. In control-treated mice, two series of OVA challenges induced upregulation of OVA-specific IgE in serum, eosinophils in the bronchoalveolar lavage fluid (BALF), and IL-5 production by lung lymphocytes upon OVA restimulation in vitro, compared with saline-challenged mice. CTLA4-IgG significantly inhibited all of these parameters in OVA-challenged mice. Importantly, mCTLA4-IgG performed better than the gold-standard dexamethasone because this corticosteroid did not inhibit the upregulation of OVA-specific IgE in serum. In a more "severe" ongoing model, induced by sensitization to OVA emulsified in aluminum hydroxide, resulting in airway hyperresponsiveness to methacholine and stronger inflammatory responses, mCTLA4-IgG was less effective in that only the number of eosinophils in the BALF was reduced (P = 0.053), whereas dexamethasone inhibited both BALF eosinophilia and cytokine production by lung lymphocytes. Thus, CTLA4-Ig might be an effective alternative therapy in established allergic asthma, especially in situations of mild disease. PMID- 11726403 TI - Association between IL-1beta/TNF-alpha-induced glucocorticoid-sensitive changes in multiple gene expression and altered responsiveness in airway smooth muscle. AB - The pleiotropic cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF) alpha have been implicated in the pathophysiology of asthma. To elucidate the role of these cytokines in the pro-asthmatic state, the effects of IL-1beta and TNF-alpha on airway smooth muscle (ASM) responsiveness and ASM expression of multiple genes, assessed by high-density oligonucleotide array analysis, were examined in the absence and presence of the glucocorticoid dexamethasone (DEX). Administration of IL-1beta/TNF-alpha increased ASM contractility to acetylcholine and impaired ASM relaxation to isoproterenol. These pro-asthmatic- like changes in ASM responsiveness were associated with IL-1beta/ TNF-alpha-induced mRNA expression of a host of proinflammatory genes that regulate transcription, cytokines and chemokines, cellular adhesion molecules, and various signal transduction molecules that regulate ASM responsiveness. In the presence of DEX, the changes induced in ASM responsiveness were abrogated, and most of the IL 1beta/TNF-alpha-mediated changes in proinflammatory gene expression were repressed, although mRNA expression of a small number of genes was enhanced by DEX. Collectively, the observations support the concept that, together with its role as a regulator of airway tone, in response to IL-1beta/TNF-alpha, the ASM expresses a host of glucocorticoid-sensitive genes that contribute to the altered structure and function of the airways in the pro-asthmatic state. We speculate that glucocorticoid-sensitive, cytokine-induced pathways involved in ASM cell signaling represent important targets for new therapeutic interventions. PMID- 11726404 TI - Cigarette smoke inhibits human bronchial epithelial cell repair processes. AB - By interfering with the ability of airway epithelial cells to support repair processes, cigarette smoke could contribute to alterations of airway structures and functions that characterize chronic obstructive pulmonary disease (COPD). The current study assessed the ability of cigarette smoke extract (CSE) to alter human airway epithelial cell chemotaxis, proliferation, and contraction of three dimensional collagen gels, a model of extracellular matrix remodeling. The volatile components contained in cigarette smoke, acetaldehyde and acrolein, were able to inhibit all three processes. Nonvolatile components contained within lyophilized CSE also inhibited chemotaxis but displayed no activity in the other two bioassays. CSE also inhibited the ability of airway epithelial cells to release transforming growth factor (TGF)-beta and fibronectin. Exogenous fibronectin was unable to restore epithelial cell contraction of collagen gels. Exogenous TGF-beta partially restored the ability of airway epithelial cells to contract collagen gels and to produce fibronectin. This supports a role for inhibition of TGF-beta release in mediating the inhibitory effects of cigarette smoke. Taken together, the results of the current study suggest that epithelial cells present in the airways of smokers may be altered in their ability to support repair responses, which may contribute to architectural disruptions present in the airways in COPD associated with cigarette smoking. PMID- 11726405 TI - Hypoxia mediates increased neutrophil and macrophage adhesiveness to alveolar epithelial cells. AB - Leukocyte infiltration is known to play an important role in hypoxia-induced tissue damage. There is a paucity of information on the role of hypoxia in the expression of adhesion molecules on respiratory epithelial cells. The current studies focus on the adhesion molecules intercellular adhesion molecule-1 (ICAM 1) and vascular cell adhesion molecule-1 (VCAM-1), their expression pattern on alveolar epithelial cells, and their biologic function under hypoxic conditions. Rat alveolar epithelial cells (AEC) were exposed to hypoxia for several time periods. With 5% oxygen, mRNA for ICAM-1 and VCAM-1 rose by 100%, peaking between 0.5 and 1 h. ICAM-1 and VCAM-1 protein showed an increase between 2 and 4 h. Neutrophil adherence to hypoxia-exposed AEC was enhanced by 115%. This increase was reduced by 83% with anti-ICAM-1 antibody. Adherence of alveolar macrophages to AEC increased by 118% and could be blocked by 95% with anti-VCAM-1 antibody. The present study shows for the first time an early increase of ICAM-1 and VCAM-1 expression on rat AEC under hypoxic conditions. These adhesion molecules are involved in increased adhesiveness of neutrophils and macrophages. Such responses might play an important role in the adhesion of leukocytes and macrophages to lung epithelial cells during hypoxic conditions. PMID- 11726407 TI - Optimizing anesthesia for inguinal herniorrhaphy: general, regional, or local anesthesia? PMID- 11726408 TI - Remifentanil for neonates and infants: piano, piano con calma. PMID- 11726409 TI - One-thousand consecutive inguinal hernia repairs under unmonitored local anesthesia. AB - To evaluate the feasibility and safety of unmonitored local anesthesia (ULA) for elective open inguinal hernia repair, we made a prospective, consecutive data collection from 1000 operations on primary and recurrent hernias. Follow-up consisted of a questionnaire 1 mo after surgery and retrieval from the electronic patient data management system. In 921 ASA Group I and II and 79 ASA Group III and IV patients, the median age was 60 yr (range, 18-95 yr). ULA was converted to general anesthesia in 5 of 1000 cases, and 961 patients were discharged on the day of surgery after 95 min (median; interquartile range, 75-150); 29 patients had complications requiring surgical intervention. Within the first month, three patients died of causes unrelated to hernia surgery, and six had cardiovascular or respiratory events. The questionnaire was returned by 940 patients; 124 were dissatisfied with local anesthesia, day-case setup, or both, primarily because of intraoperative pain (n = 74; 7.8%). We conclude that open inguinal hernia repair can be conducted under ULA, regardless of comorbidity, with a small rate of deviation from day-case setup and minimal morbidity. It provides a safe alternative to other anesthetic techniques with an acceptable rate of satisfaction, but intraoperative pain relief needs improvement. IMPLICATIONS: Inguinal hernia repair can be safely performed under unmonitored local anesthesia with infrequent postoperative morbidity and acceptable satisfaction, but intraoperative pain may be a problem. PMID- 11726410 TI - Selective spinal anesthesia: a comparison of hyperbaric bupivacaine 4 mg versus 6 mg for outpatient knee arthroscopy. AB - IMPLICATIONS: A low-dose (4 mg), low-volume (0.8 mL), low-flow (2 min) technique with hyperbaric bupivacaine toward the dependent side oriented injection and maintenance of the lateral decubitus position for 10 min produced selective spinal anesthesia with rapid recession of motor block and early discharge home. PMID- 11726411 TI - A randomized multicenter study of remifentanil compared with halothane in neonates and infants undergoing pyloromyotomy. I. Emergence and recovery profiles. AB - Pyloric stenosis is sometimes associated with hemodynamic instability and postoperative apnea. In this multicenter study we examined the hemodynamic response and recovery profile of remifentanil and compared it with that of halothane in infants undergoing pyloromyotomy. After atropine, propofol, and succinylcholine administration and tracheal intubation, patients were randomized (2:1 ratio) to receive either remifentanil with nitrous oxide and oxygen or halothane with nitrous oxide and oxygen as the maintenance anesthetic. Pre- and postoperative pneumograms were done and evaluated by an observer blinded to the study. Intraoperative hemodynamic data and postanesthesia care unit (PACU) discharge times, PACU recovery scores, pain medications, and adverse events (vomiting, bradycardia, dysrhythmia, and hypoxemia) were recorded by the study's research nurse. There were no significant differences in patient age or weight between the two groups. There were no significant differences in hemodynamic values between the two groups at the various intraoperative stress points. The extubation times, PACU discharge times, pain medications, and adverse events were similar for both groups. No patient anesthetized with remifentanil who had a normal preoperative pneumogram had an abnormal postoperative pneumogram, whereas three patients with a normal preoperative pneumogram who were anesthetized with halothane had abnormal pneumograms after. IMPLICATIONS: The use of ultra-short acting opioids may be an appropriate technique for infants less than 2 mo old when tracheal extubation after surgery is anticipated. PMID- 11726412 TI - A randomized multicenter study of remifentanil compared with halothane in neonates and infants undergoing pyloromyotomy. II. Perioperative breathing patterns in neonates and infants with pyloric stenosis. AB - Although former preterm birth infants are at risk for postoperative apnea after surgery, it is unclear whether the same is true of full-term birth infants. We evaluated the incidence of apnea in 60 full-term neonates and infants undergoing pyloromyotomy both before and after anesthesia. All subjects were randomized to a remifentanil- or halothane-based anesthetic. Apnea was defined by the presence of prolonged apnea (>15 s) or frequent brief apnea, as observed on the pneumocardiogram. Apnea occurred before surgery in 27% of subjects and after surgery in 16% of subjects, with no significant difference between subjects randomized to remifentanil or halothane anesthesia. This apnea was primarily central in origin, occurred throughout the recording epochs, and was associated with severe desaturation in some instances. Of the subjects with normal preoperative pneumocardiograms, new onset postoperative apnea occurred in 3 (23%) of 13 subjects who received halothane-based anesthetics versus 0 (0%) of 22 subjects who received remifentanil-based anesthetics (P = 0.04). Thus, postoperative apnea can follow anesthesia in otherwise healthy full-term infants after pyloromyotomy and is occasionally severe with desaturation. New-onset postoperative apnea was not seen with a remifentanil-based anesthetic. IMPLICATIONS: Abnormal breathing patterns can follow anesthesia in infants after surgical repair of pyloric stenosis. Occasionally, these patterns can be associated with desaturation. New-onset postoperative apnea was not seen with a remifentanil-based anesthetic. PMID- 11726413 TI - Pharmacokinetics of remifentanil in anesthetized pediatric patients undergoing elective surgery or diagnostic procedures. AB - Remifentanil hydrochloride is an ultra-short-acting opioid that undergoes rapid metabolism by tissue and plasma esterases. We aimed to characterize the pharmacokinetics and determine the hemodynamic profile of remifentanil after a single-bolus dose in children aged 0 to 18 yr. Forty-two children undergoing elective surgical procedures received remifentanil 5 microg/kg infused over 1 min. Patients were divided into age groups as follows: young infants (< or =2 mo), older infants (> 2 mo to < 2 yr), young children (2 to < 7 yr), older children (7 to < 13 yr), adolescents (13 to < 16 yr), and young adults (16 to < 18 yr). Arterial blood samples were collected and analyzed by mass spectroscopy to determine remifentanil pharmacokinetic profiles. Hemodynamic measurements for remifentanil's effect were made after the infusion. Methods of statistical analysis included analysis of variance and linear regression, with significance at P < or = 0.05. Complete remifentanil pharmacokinetic data were obtained from 34 patients. The volume of distribution was largest in the infants < 2 mo (mean, 452 mL/kg) and decreased to means of 223 to 308 mL/kg in the older patients. There was a more rapid clearance in the infants < 2 mo of age (90 mL. kg(-1). min(-1)) and infants 2 mo to 2 yr (92 mL. kg(-1). min(-1)) than in the other groups (means, 46 to 76 mL. kg(-1). min(-1)). The half-life was similar in all age groups, with means of 3.4 to 5.7 min. Seven subjects (17%) developed hypotension related to the remifentanil bolus. Remifentanil showed an extremely rapid elimination similar to that in adults. The fast clearance rates observed in neonates and infants, as well as the lack of age-related changes in half-life, are in sharp contrast to the pharmacokinetic profile of other opioids. Remifentanil in a bolus dose of 5 microg/kg may cause hypotension in anesthetized children. IMPLICATIONS: The pharmacokinetics of remifentanil were studied in children from birth to 18 yr. Remifentanil was found to have age-related changes in clearance and volume of distribution, but not half-life. The increased clearance observed in young infants is in contrast to other opioids. PMID- 11726414 TI - The continuous recording of blood pressure in patients undergoing carotid surgery under remifentanil versus sufentanil analgesia. AB - We compared the hemodynamic stability during carotid endarterectomy of remifentanil with that of sufentanil anesthesia. Fifty-six patients were randomly assigned into Remifentanil (n = 27) or Sufentanil (n = 29) groups. In the Remifentanil group, IV propacetamol (2 g) and morphine (0.1 mg/kg) were infused 30 min before skin closure. In the Sufentanil group, patients received 2 g propacetamol. Beat-to-beat recordings of systolic arterial blood pressure (SBP) and heart rate (HR) were stored on a computer. The maximum and minimum values of BP and HR after induction, at intubation, during the surgical procedure, and after the operation and the coefficients of variation of SBP and HR were used as indices of hemodynamic stability. The coefficients of variation of SBP and HR were similar in both groups during and after surgery. However, at intubation, maximal SBP was higher in the Sufentanil group (P < 0.05). Decreased propofol doses and isoflurane end-tidal concentrations were used in the Remifentanil group. At recovery, a similar profile of SBP and HR was found in both groups. We conclude that intra- and posthemodynamic stability was similar with remifentanil or sufentanil in patients undergoing carotid endarterectomy. However, remifentanil was more effective for blunting the increase in SBP at intubation without increasing the blood pressure-decreasing effect of induction. Intraoperative remifentanil use was associated with a decreased amount of hypnotic drug administered. IMPLICATIONS: Beat-to-beat recordings of heart rate and blood pressure in patients undergoing carotid surgery revealed that hemodynamic stability was similar with remifentanil or sufentanil anesthesia both during and after surgery. Remifentanil was more effective in limiting the increase in blood pressure associated with intubation without increasing the blood pressure-lowering effect of induction or the blood pressure response to recovery. PMID- 11726415 TI - Risk factors for ischemic optic neuropathy after cardiopulmonary bypass: a matched case/control study. AB - Visual loss (acuity or field) secondary to ischemic optic neuropathy (ION) is a rare but devastating complication of cardiac surgery involving cardiopulmonary bypass (CPB). We determined clinical features and risk factors for ION by a retrospective time-matched, case-control study. ION was identified in 17 (0.06%) patients out of 27,915 patients who underwent CPB between January 1, 1976, and December 31, 1994. For each ION patient, two patients who underwent CPB exactly 2 wk before the ION patient were selected as controls. Data were analyzed by using conditional logistic regression with the 1:2 matched-set feature of 17 cases and 34 controls. Two-tailed P values < or =0.05 were considered significant. From bivariate analysis, smaller minimum postoperative hemoglobin concentration (odds ratio [OR] = 1.9, P = 0.047) and the presence of atherosclerotic vascular disease (OR = 7.0, P = 0.026) were found to be independently associated with ION after CPB, as were smaller minimum postoperative hemoglobin concentration (OR = 2.2, P = 0.027) and preoperative angiogram within 48 h of surgery (OR = 7.2, P = 0.042). In ION patients, 13 (76.5%) of 17 experienced a minimum postoperative hemoglobin value of < 8.5 g/dL, whereas only 14 (41.2%) of 34 control patients experienced values < 8.5 g/dL. IMPLICATIONS: Patients with clinically significant vascular disease history or preoperative angiogram may be at increased risk for ischemic optic neuropathy after cardiac surgery, especially if the hemoglobin remains low in the postoperative period. PMID- 11726416 TI - The frequency of perioperative vision loss. AB - The frequency of perioperative vision loss, especially for spinal surgery, has been increasing recently. We undertook a retrospective study to determine the frequency of this outcome in a large surgical population receiving general or central neuraxis regional anesthesia for noncardiac procedures from 1986 to 1998. Specific criteria were used to separate cases in which the surgical procedure likely directly contributed to the vision loss. Vision loss was present if any part of the visual field was affected. Initial database screening found 405 cases of new-onset vision loss or visual changes in 410,189 patients who underwent 501,342 anesthetics and who survived at least 30 days after their final procedures. Two hundred sixteen of these patients regained full vision or acuity within 30 days. Of the 189 patients who developed vision deficits for longer than 30 days, 185 underwent ophthalmologic or neurologic procedures in which ocular or cerebral tissues were surgically damaged or resected. The remaining 4 patients (1 per 125,234 overall; 0.0008%) developed prolonged vision loss without direct surgical trauma to optic or cerebral tissues. In this large study population of noncardiac surgical patients, including those who underwent spinal surgical procedures, the frequency of perioperative vision loss persisting for longer than 30 days was very small. IMPLICATIONS: Vision loss and blindness after surgery and anesthesia is a very rare event. In this study, only one per 125,234 patients undergoing noncardiac surgery developed vision loss persisting for longer than 30 days. PMID- 11726417 TI - Training and certification in perioperative transesophageal echocardiography: a historical perspective. PMID- 11726418 TI - The predictive value of ventricular fibrillation electrocardiogram signal frequency and amplitude variables in patients with out-of-hospital cardiac arrest. AB - We evaluated ventricular fibrillation frequency and amplitude variables to predict successful countershock, defined as pulse-generating electrical activity. We also elucidated whether bystander cardiopulmonary resuscitation (CPR) influences these electrocardiogram (ECG) variables. In 89 patients with out-of hospital cardiac arrest, ECG recordings of 594 countershock attempts were collected and analyzed retrospectively. By using fast Fourier transformation analysis of the ventricular fibrillation ECG signal in the frequency range 0.333 15 Hz (median [range]), median frequency, dominant frequency, spectral edge frequency, and amplitude were as follows: 4.4 (2.4-7.5) Hz, 4.0 (0.7-7.0) Hz, 7.7 (3.7-13.7) Hz, and 0.94 (0.24-1.95) mV, respectively, before successful countershock (n = 59). These values were 3.8 (0.8-7.7) Hz (P = 0.0002), 3.0 (0.3 9.7) Hz (P < 0.0001), 7.3 (2.0-14.0) Hz (P < 0.05), and 0.53 (0.03-3.03) mV (P < 0.0001), respectively, before unsuccessful countershock (n = 535). In patients in whom bystander CPR was performed (n = 51), ventricular fibrillation frequency and amplitude before the first defibrillation attempt were higher than in patients without bystander CPR (n = 38) (median frequency, 4.4 [2.4-7.5] vs 3.7 [1.8-5.3] Hz, P < 0.0001; dominant frequency, 3.8 [0.9-7.7] vs 2.6 [0.8-5.9] Hz, P < 0.0001; spectral edge frequency, 8.4 [4.8-12.9] vs 7.2 [3.9-12.1] Hz, P < 0.05; amplitude, 0.79 [0.06-4.72] vs 0.67 [0.16-2.29] mV, P = 0.0647). Receiver operating characteristic curves demonstrate that successful countershocks will be best discriminated from unsuccessful countershocks by ventricular fibrillation amplitude (3000-ms epoch). At 73% sensitivity, a specificity of 67% was obtained with this variable. IMPLICATIONS: In patients with out-of-hospital cardiac arrest, successful countershocks will be best discriminated from unsuccessful countershocks by ventricular fibrillation amplitude (3000-ms epoch). At 73% sensitivity, a specificity of 67% was obtained with this variable. PMID- 11726419 TI - The effects of increasing concentrations of isoflurane and desflurane on pulmonary perfusion and systemic oxygenation during one-lung ventilation in pigs. AB - During one-lung ventilation (OLV), hypoxic pulmonary vasoconstriction (HPV) reduces venous admixture and attenuates the decrease in arterial oxygen tension by diverting blood from the nonventilated lung to the ventilated lung. In vitro, desflurane and isoflurane depress HPV in a dose-dependent manner. Accordingly, we studied the effects of increasing concentrations of desflurane and isoflurane on pulmonary perfusion, shunt fraction, and PaO(2) during OLV in vivo. Fourteen pigs (30-42 kg) were anesthetized, tracheally intubated, and mechanically ventilated. After placement of femoral arterial and thermodilution pulmonary artery catheters, a left-sided double-lumen tube (DLT) was placed via tracheotomy. After DLT placement, FIO(2) was adjusted at 0.8 and anesthesia was continued in random order with 3 concentrations (0.5, 1.0, and 1.5 minimal alveolar concentrations) of either desflurane or isoflurane. Differential lung perfusion was measured with colored microspheres. All measurements were made after stabilization at each concentration. Whereas mixed venous PO(2), mean arterial pressure, cardiac output, nonventilated lung perfusion, and shunt fraction decreased in a dose dependent manner, PaO(2) remained unchanged with increasing concentrations of desflurane and isoflurane during OLV. In conclusion, increasing concentration of desflurane and isoflurane did not impair oxygenation during OLV in pigs. IMPLICATIONS: In an animal model of one-lung ventilation, increasing concentrations of desflurane and isoflurane dose-dependently decreased shunt fraction and perfusion of the nonventilated lung and did not impair oxygenation. The decreases in shunt fraction are likely the result of anesthetic-induced marked decreases in cardiac output and mixed venous saturation. PMID- 11726420 TI - The additive pulmonary vasodilatory effects of inhaled prostacyclin and inhaled milrinone in postcardiac surgical patients with pulmonary hypertension. AB - Selective pulmonary vasodilation is an advantageous therapeutic strategy for cardiac surgical patients with increased pulmonary vascular resistance (PVR) and right ventricular failure. We hypothesized that milrinone, an adenosine-3',5' cyclic monophosphate (cAMP)-selective phosphodiesterase enzyme (PDE) inhibitor may, when nebulized and inhaled, cause selective pulmonary vasodilation and potentiate the vasodilation by inhaled prostacyclin (iPGI(2)). Consequently, we investigated the hemodynamic effects of inhaled milrinone or the combination iPGI(2) + inhaled milrinone in cardiac surgical patients with postoperative mean pulmonary arterial pressure (MPAP) >25 mm Hg and PVR >200 dynes. s(-1). cm(-5). During mechanical ventilation and using a conventional nebulizing system, 9 patients inhaled incremental concentrations of milrinone (0.25, 0.5 and 1 mg/mL) in subsequent 10-min periods (Study Part 1). In the same manner, 11 patients received iPGI(2) (10 microg/mL) followed by the combination of iPGI(2) (10 microg/mL) and inhaled milrinone (1 mg/mL) (Study Part 2). Inhaled milrinone reduced PVR with a maximal effect (-20%, P < 0.001) at the largest concentration. As compared with iPGI(2) alone, iPGI(2) + inhaled milrinone caused a further and prolonged reduction of PVR (-8%, P < 0.05) and increased stroke volume (+5%, P < 0.05). Systemic vascular resistance or mean arterial pressure was not affected by inhalation of either drug(s). The authors conclude that inhalation of the cAMP selective PDE-inhibitor milrinone selectively dilates the pulmonary vasculature without systemic effects in cardiac surgical patients with pulmonary hypertension. Furthermore, inhaled milrinone appears to potentiate and prolong the pulmonary selective vasodilatory effect of iPGI(2). Inhaled milrinone alone or combined with iPGI(2) may be an important therapeutic option in the treatment of patients with pulmonary hypertension and right ventricular failure. IMPLICATIONS: Pulmonary hypertension may cause or aggravate right heart failure. IV vasodilators reduce systemic blood pressure and might thereby further impair coronary perfusion and right heart performance. In the present study of cardiac surgical patients with pulmonary hypertension, selective pulmonary vasodilation without systemic effects was induced by nebulized, inhaled vasodilators. PMID- 11726421 TI - A dose-determining trial of heparinase-I (Neutralase) for heparin neutralization in coronary artery surgery. AB - Heparinase-I, a specific heparin-degrading enzyme, may represent an alternative to protamine. We explored the dose of heparinase-I for efficacy and safety in patients undergoing coronary artery surgery. At the conclusion of cardiopulmonary bypass, subjects received 5, 7, or 10 microg/kg of open-label heparinase-I instead of protamine. Activated clotting time (ACT) and its difference from a contemporaneous heparin-free sample (DeltaACT) at 3 min before and 3, 6, and 9 min after heparinase-I determined reversal efficacy. After surgery, we recorded hourly chest tube drainage. Systemic and pulmonary arterial blood pressure and cardiac output measurements before and immediately after heparinase-I were used to evaluate hemodynamic safety. Coagulation measurements included anti-factor Xa and anti-factor IIa activities. Forty-nine patients from seven institutions participated: 12 received 5 microg/kg, 21 received 7 microg/kg, 4 received two doses of 7 microg/kg, 8 received 10 microg/kg, and 4 received two doses of 10 microg/kg. Treatment groups did not differ demographically. Median DeltaACT 9 min later was 11, 7, and 4 s for the 5, 7, and 10 microg/kg groups, respectively. No adverse hemodynamic changes occurred with heparinase-I administration. The authors conclude that heparinase-I effectively restored the ACT after cardiopulmonary bypass. This effect appeared to be dose dependent. IMPLICATIONS: Heparinase-I (Neutralase(TM)) successfully restored activated coagulation time with no adverse hemodynamic events in patients undergoing coronary artery surgery with cardiopulmonary bypass in an open-label dose-determining trial. PMID- 11726422 TI - The in vitro reversal of histamine-induced vasodilation in the human internal mammary artery. AB - Anaphylactic shock therapy includes the use of catecholamines but they may not always be effective. Because vasodilation during anaphylaxis is a result of the endothelial release of multiple mediators, we investigated the effects of epinephrine, vasopressin, and inhibitors of nitric oxide and prostanoid pathways on histamine-induced relaxation in human internal mammary artery. The vessel segments were obtained intraoperatively and were suspended in organ chambers to record isometric tension. Norepinephrine (10(-6) M) was used to precontract the rings followed by histamine (10(-6.5) M) to relax the vessels and mimic vascular collapse. Epinephrine, vasopressin, methylene blue, N(G)-monomethyl-L-arginine (L NMA) and indomethacin were added in a cumulative fashion to reverse the histamine induced vasodilation. The internal mammary artery segments exhibited greater contraction in the presence of the epinephrine (4.9 +/- 0.7 g) compared with vasopressin (2.6 +/- 0.7 g). Vasopressin (10(-11) to 10(-7) M), methylene blue (10(-7) to 10(-5) M), L-NMA (10(-6) to 10(-4) M), and indomethacin (10(-7) to 10( 5) M) were only partially effective. These findings suggest that vasopressin and methylene blue may offer a potential therapeutic option in the treatment of histamine-induced vasodilatory shock. IMPLICATIONS: Epinephrine only partially reverses histamine-induced vasodilation in human internal mammary arteries, whereas vasopressin, methylene blue, and drugs involved in the inhibition of nitric oxide and prostaglandin generation lead to a complete reversal of the vascular relaxation. PMID- 11726423 TI - Thoracic, but not lumbar, epidural anesthesia improves cardiopulmonary function in ovine pulmonary embolism. AB - We hypothesized that sympathetic stimulation is the main mechanism contributing to hemodynamic failure in pulmonary embolism. We investigated the effects of epidural anesthesia-induced sympathetic blockade, restricted to thoracic and lumbar levels, during pulmonary embolism. Two experiments were performed in chronically instrumented ewes. In the first experiment, six sheep received 6 mL bupivacaine 0.175% (Thoracic Epidural Anesthesia [TEA] group), and six sheep received 6 mL saline 0.9% (TEA-Control group), respectively, via an epidural catheter (T3 level). In the second experiment, six sheep received 2.8 mL bupivacaine 0.375% (Lumbar Epidural Anesthesia [LEA] group), and six sheep received 2.8 mL saline 0.9% (LEA-Control group) epidurally (L4 level). Embolization was performed by IV injection of autologous blood clots (Experiment 1, 0.75 mL/kg; Experiment 2, 0.625 mL/kg). TEA was associated with significantly slower heart rates, decreased mean pulmonary artery pressures and central venous pressures, and significantly higher stroke volume index and oxygenation in comparison with the TEA-Control group. By contrast, LEA was associated with significantly faster heart rates and increased central venous pressures and with a significantly lower stroke volume index in comparison with the LEA-Control group. TEA significantly reduced, and LEA significantly increased, hemodynamic deterioration, suggesting beneficial effects of TEA on cardiopulmonary function during pulmonary thromboembolism. IMPLICATIONS: Thoracic (but not lumbar) epidural anesthesia was associated with beneficial cardiopulmonary effects during experimental pulmonary thromboembolism in sheep. PMID- 11726424 TI - How does the plethysmogram derived from the pulse oximeter relate to arterial blood pressure in coronary artery bypass graft patients? AB - Twenty patients scheduled for coronary artery bypass grafting had their ear and finger oximeter and radial artery blood pressure (Bp(meas)) waveforms collected. The ear and finger pulse oximeter waveforms were analyzed to extract beat-to-beat amplitude and area and width measurements. The Bp(meas) waveforms were analyzed to measured systolic blood pressure (BP), mean BP, and pulse pressure. The correlation coefficient was determined between the derived waveforms from the pulse oximeter and Bp(meas) for the first 10 patients. The ear pulse oximeter width (Width(Ear)) had the best correlation (r = 0.8). Linear regression was done between Width(Ear) and Bp(meas) based on slope (b) and intercept (a) values, BP was calculated (Bp(calc)) in the next 10 patients as: [equation: see text] where i = systolic BP, mean BP, and pulse pressure. The initial bias was too large to be clinically useful. To improve clinical applicability a period of calibration was introduced in which the first 50 readings of Width(Ear) and Bp(meas) for each patient were used to calculate the intercept. After calibration the systolic BP, mean BP and pulse pressure bias values were -2.6, -1.88 and -1.28 mm Hg, and the precision values were 15.9 10.09, and 9.94 mm Hg, respectively. The present attempt to develop a clinically useful method of noninvasive BP measuring was partly successful with the requirement of a calibration period. IMPLICATIONS: Statistical comparison was made between measured blood pressure (BP) from arterial line and calculated BP derived from ear pulse oximeter waveform in 10 patients undergoing coronary artery bypass graft surgery. Using 62,077 paired readings, the mean difference for systolic BP, mean BP, and pulse pressure between the 2 methods was -2.6, -1.88, and -1.28 mm Hg, respectively. PMID- 11726425 TI - Cariporide (HOE 642) attenuates leukocyte activation in ischemia and reperfusion. AB - Cariporide (HOE 642) ameliorates myocardial ischemia/reperfusion (I/R) injury, by the well established reduction of cytosolic [Ca(2+)] in cardiac myocytes through inhibition of Na(+)/H(+) exchange. However, postischemic inflammation also contributes to I/R injury. We tested the hypothesis that cariporide also modulates the inflammatory response. The effect of cariporide on L-selectin expression by human leukocytes in vitro and leukocyte adhesion and emigration in the reperfused rat cremaster muscle in vivo were studied. The rat cremaster muscle was exteriorized for intravital videomicroscopy, induction of ischemia (90 min), and reperfusion (90 min). Eleven rats were pretreated with cariporide (9 mg/kg body weight IV) whereas 11 rats received saline. Leukocyte adhesion was quantified offline. Human venous blood was incubated with cariporide (3 micromol/L) or saline, stimulated with formyl- methionine-leucine-phenylalanine (10(-10)-10(-6) mol/L), and granulocyte L-selectin expression was analyzed by flow cytometry. Cariporide reduced leukocyte rolling and adhesion by approximately 35% and 45%, respectively, after 30 min of reperfusion. Leukocyte extravasation was decreased by approximately 85% after 90 min. Cariporide increased L-selectin shedding at each formyl-methionine-leucine-phenylalanine concentration, reducing the 50% effective dose from 9.95 to 4.68 nmol/L. Thus, cariporide may ameliorate I/R injury not only by the known reduction of cytosolic [Ca(2+)] in cardiomyocytes, but also by attenuating leukocyte-dependent inflammatory responses. Promotion of L-selectin shedding from activated leukocytes may present a mechanism underlying this newly detected effect. IMPLICATIONS: This study provides evidence that inhibition of Na(+)/H(+) exchange by cariporide (HOE 642) attenuates the postischemic inflammatory response. Leukocyte adhesion and emigration, assessed by in vivo microscopy, were markedly reduced in rat cremaster muscle, possibly because of increased L-selectin shedding of activated leukocytes as demonstrated by flow cytometry. PMID- 11726426 TI - Hemostatic analysis of a patient with hereditary angioedema undergoing coronary artery bypass grafting. AB - IMPLICATIONS: Hereditary angioedema is a disease associated with acute complement mediated inflammation and swelling of the airway and other vital organs. This case describes the impact of hereditary angioedema and cardiopulmonary bypass on hemostasis as assessed by thrombelastography. PMID- 11726427 TI - Errors during intraoperative cell salvage because of inappropriate wash solutions. AB - IMPLICATIONS: Two case reports illustrate errors that can occur during intraoperative red blood cell salvage and emphasize the need for standardized procedures and quality improvement processes for this intervention. PMID- 11726428 TI - Minimally invasive direct coronary artery bypass surgery under high thoracic epidural. AB - IMPLICATIONS: This report describes the use of high-thoracic epidural anesthesia for a patient undergoing minimally invasive direct coronary artery bypass. PMID- 11726429 TI - The recovery of cognitive function after general anesthesia in elderly patients: a comparison of desflurane and sevoflurane. AB - We evaluated the cognitive recovery profiles in elderly patients after general anesthesia with desflurane or sevoflurane. After IRB approval, 70 ASA physical status I-III consenting elderly patients (> or =65 yr old) undergoing total knee or hip replacement procedures were randomly assigned to one of two general anesthetic groups. Propofol and fentanyl were administered for induction of anesthesia, followed by either desflurane 2%-4% or sevoflurane 1%-1.5% with nitrous oxide 65% in oxygen. The desflurane (2.5 +/- 0.6 MAC. h) and sevoflurane (2.7 +/- 0.5 MAC. h) concentrations were adjusted to maintain comparable depths of hypnosis using the electroencephalogram bispectral index monitor. The Mini Mental State (MMS) test was used to assess cognitive function preoperatively and postoperatively at 1, 3, 6, and 24-h intervals. The use of desflurane was associated with a more rapid emergence from anesthesia (6.3 +/- 2.4 min versus 8.0 +/- 2.8 min) and a shorter length of stay in the postanesthesia care unit (213 +/- 66 min versus 241 +/- 87 min). However, there were no significant differences between the Desflurane and the Sevoflurane groups when the MMS scores were compared preoperatively, and postoperatively at 1, 3, 6, and 24 h. Compared with the preoperative (baseline) MMS scores, the values were significantly decreased at 1 h postoperatively (27.8 +/- 1.7 versus 29.5 +/- 0.5 in the Desflurane group, and 27.4 +/- 1.7 versus 29.2 +/- 1.0 in the Sevoflurane group, respectively). However, the MMS scores returned to preoperative baseline levels within 6 h after surgery. At 1 h and 3 h after surgery, 51% and 11% (versus 57% and 9%) of patients in the Desflurane (versus Sevoflurane) Group experienced cognitive impairment. In conclusion, desflurane is associated with a faster early recovery than sevoflurane after general anesthesia in elderly patients. However, recovery of cognitive function was similar after desflurane and sevoflurane-based anesthesia. IMPLICATIONS: Desflurane was associated with a faster early recovery than sevoflurane after general anesthesia in elderly patients. However, recovery of cognitive function was similar with both volatile anesthetics. PMID- 11726430 TI - The involvement of the mu-opioid receptor in ketamine-induced respiratory depression and antinociception. AB - N-methyl-D-aspartate receptor antagonism probably accounts for most of ketamine's anesthetic effects; its analgesic properties are mediated partly via N-methyl-D aspartate and partly via opioid receptors. We assessed the involvement of the mu opioid receptor in S(+) ketamine-induced respiratory depression and antinociception by performing dose-response curves in exon 2 mu-opioid receptor knockout mice (MOR(-/-)) and their wild-type littermates (WT). The ventilatory response to increases in inspired CO(2) was measured with whole body plethysmography. Two antinociceptive assays were used: the tail-immersion test and the hotplate test. S(+) ketamine (0, 10, 100, and 200 mg/kg intraperitoneally) caused a dose-dependent respiratory depression in both genotypes, with greater depression observed in WT relative to MOR(-/-) mice. At 200 mg/kg, S(+) ketamine reduced the slope of the hypercapnic ventilatory response by 93% +/- 15% and 49% +/- 6% in WT and MOR(-/-) mice, respectively (P < 0.001). In both genotypes, S(+) ketamine produced a dose-dependent increase in latencies in the hotplate test, with latencies in MOR(-/-) mice smaller compared with those in WT animals (P < 0.05). In contrast to WT mice, MOR(-/-) mice displayed no ketamine-induced antinociception in the tail-immersion test. These results indicate that at supraspinal sites S(+) ketamine interacts with the mu opioid system. This interaction contributes significantly to S(+) ketamine induced respiratory depression and supraspinal antinociception. IMPLICATIONS: The involvement of the mu-opioid receptor system in S(+) ketamine-induced respiratory depression and spinal and supraspinal analgesia was demonstrated by performing experiments in mice lacking the mu-opioid receptor and in mice with intact mu opioid receptors. PMID- 11726431 TI - The effects of lidocaine on superoxide production and p47 Phox translocation in opsonized zymosan-activated neutrophils. AB - Superoxide is generated by reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase that exists in the cell membrane of neutrophils and contains p47 phox in the enzyme complex. Superoxide is closely related to various diseases, including infection, myocardial infarction, and aspiration pneumonia. Therefore we performed this study to establish how lidocaine acts on NADPH oxidase. We examined the effects of lidocaine on superoxide production in neutrophils using the method of 2-methyl-6-phenyl-3,7-dihydro [1,2-a] pyrazin-3-one (CLA phenyl) dependent chemiluminescence activated by opsonized zymosan (OZ). Treatment with lidocaine 15 microM suppressed OZ-activated CLA phenyl-dependent chemiluminescence by approximately 40% compared with that of OZ only-activated neutrophils. It was further demonstrated that the suppressive effect of lidocaine on superoxide production was dependent on the concentration of lidocaine. Translocation of p47 phox from the cytosol to the cell membrane was studied using immunoblot analysis with a polyclonal anti-p47 phox antibody. Lidocaine suppressed OZ-induced p47 phox translocation in a dose-dependent manner. Our results suggest that suppression of superoxide production by a therapeutic dose of lidocaine correlates strongly with suppression of p47 phox translocation. IMPLICATIONS: We report on the effects of lidocaine on superoxide production in opsonized zymosan-activated neutrophils. Lidocaine suppresses superoxide production in a dose-dependent manner. This suppression correlates strongly with the suppression of translocation of p47 phox, a new subunit of NADPH-oxidase. PMID- 11726432 TI - Preoperative oral B vitamins prevent nitrous oxide-induced postoperative plasma homocysteine increases. AB - Nitrous oxide increases total homocysteine (tHcy) plasma levels, which are associated with an increase in perioperative myocardial ischemia. We designed this study to determine whether oral B vitamins, which are cofactors in homocysteine metabolism, can prevent nitrous oxide anesthesia-induced tHcy increases in patients undergoing elective surgery scheduled to last longer than 3 h. Fifty-three patients presenting for elective revision knee or hip arthroplasty received in random, double-blinded fashion oral vitamin B complex (folate 2.5 mg, B(6) 25 mg, and B(12) 500 microg) or placebo daily for 1 wk before surgery. Anesthesia was induced with propofol and maintained with an opioid, isoflurane, and nitrous oxide/oxygen (inspired nitrous oxide >50%). Blood samples for measurement of tHcy concentration were obtained at study enrollment, before induction, on arrival in the postanesthesia care unit, and on Day 5. Fourteen patients had their surgery rescheduled after taking their vitamins and were removed from the study. The Placebo group had a mean increase in tHcy concentration from baseline of 15% +/- 31% compared with the Vitamin group, which had an initial decrease of 9.1% +/- 11% (P = 0.035). This was maintained throughout the 5-day study period. The use of an oral B vitamin complex prevented the increase in postoperative tHcy by nitrous oxide. IMPLICATIONS: The use of nitrous oxide anesthesia increases postoperative homocysteine concentrations and associated myocardial ischemia. This study indicates that a 1-wk course of oral B vitamins can prevent the increase in homocysteine from nitrous oxide, and, by implication, myocardial ischemia as well. PMID- 11726433 TI - Long-duration low-flow sevoflurane and isoflurane effects on postoperative renal and hepatic function. AB - Sevoflurane degradation by carbon dioxide absorbents during low-flow anesthesia forms the haloalkene Compound A, which causes nephrotoxicity in rats. Numerous studies have shown no effects of Compound A formation on postoperative renal function after moderate-duration (3-4 h) low-flow sevoflurane; however, effects of longer exposures remain unresolved. We compared renal function after long duration low-flow (<1 L/min) sevoflurane and isoflurane anesthesia in consenting surgical patients with normal renal function. To maximize degradant exposure, Baralyme was used, and anesthetic concentrations were maximized (no nitrous oxide and minimal opioids). Inspired and expired Compound A concentrations were quantified. Blood and urine were obtained for laboratory evaluation. Sevoflurane (n = 28) and isoflurane (n = 27) groups were similar with respect to age, sex, weight, ASA status, and anesthetic duration (9.1 +/- 3.0 and 8.2 +/- 3.0 h, mean +/- SD) and exposure (9.2 +/- 3.6 and 9.1 +/- 3.7 minimum alveolar anesthetic concentration hours). Maximum inspired Compound A was 25 +/- 9 ppm (range, 6-49 ppm), and exposure (area under the concentration-time curve) was 165 +/- 95 (35 428) ppm. h. There was no significant difference between anesthetic groups in 24- or 72-h serum creatinine, blood urea nitrogen, creatinine clearance, or 0- to 24 h or 48- to 72-h urinary protein or glucose excretion. Proteinuria and glucosuria were common in both groups. There was no correlation between Compound A exposure and any renal function measure. There was no difference between anesthetic groups in 24- or 72-h aspartate aminotransferase or alanine aminotransferase. These results show that the renal and hepatic effects of long-duration low-flow sevoflurane and isoflurane were similar. No evidence for low-flow sevoflurane nephrotoxicity was observed, even at high Compound A exposures as long as 17 h. Proteinuria and glucosuria were common and nonspecific postoperative findings. Long-duration low-flow sevoflurane seems as safe as long-duration low-flow isoflurane anesthesia. IMPLICATIONS: Postoperative renal function after long duration low-flow sevoflurane (with Compound A exposures greater than those typically reported) and isoflurane anesthesia were not different, as assessed by serum creatinine, blood urea nitrogen, and urinary excretion of protein and glucose. This suggests that low-flow sevoflurane is as safe as low-flow isoflurane, even at long exposures. PMID- 11726435 TI - Esmolol promotes electroencephalographic burst suppression during propofol/alfentanil anesthesia. AB - This study examined the effects of an esmolol infusion on the electroencephalogram during propofol/alfentanil IV anesthesia. After informed consent, 20 patients were randomly assigned into four groups on the basis of two target alfentanil concentrations (alfentanil 50 or 150 ng/mL) and of a saline or esmolol infusion. Bispectral index (BIS), burst suppression ratio (SR), and physiologic variables were continuously monitored. A 30-min blinded infusion of saline or esmolol was started after establishing a stable baseline and followed by a washout period. The electroencephalogram was significantly suppressed by esmolol (BIS, 37 +/- 6 to 22 +/- 6, 40% decrease [mean +/- SD]; SR, 5 +/- 7 to 67 +/- 23, 13.4-fold increase) compared with baseline in the small-dose alfentanil groups. Discontinuation of esmolol reversed the response. BIS and SR were unaffected by placebo infusion. Twelve-minute to 16-min hysteresis between esmolol administration and the onset of half-maximal cortical suppression was observed. Physiologic variables and serum propofol and alfentanil concentrations were not significantly altered by esmolol. Although the mechanism remains unclear, significant cortical depression and the onset of burst suppression during a stable, computer-controlled propofol/alfentanil anesthetic was associated with esmolol infusion. IMPLICATIONS: This study demonstrated the suppression of cerebral cortical electrical activity after blinded esmolol infusion during propofol/alfentanil anesthesia. A significant lag was noted between infusion and half-maximal effect (12-16 min). Whether esmolol, a metabolite, or a secondary process was responsible for this cortical suppression remains unknown and requires further study. PMID- 11726434 TI - Small-dose pentobarbital enhances synaptic transmission in rat hippocampus. AB - We investigated the contribution of bicarbonate ion, gamma-aminobutyric acid-A (GABA(A)) receptors, and N-methyl-D-aspartate (NMDA) receptors to pentobarbital induced enhancement of excitatory synaptic transmission in the hippocampal slice. Transverse hippocampal slices (400 microm thick) were prepared from 20- to 30-day old Sprague-Dawley rats and maintained in an interface chamber perfused with warmed (35 degrees C) oxygenated artificial cerebrospinal fluid. Extracellular field potentials, evoked by orthodromic paired-pulse stimulation of the Schaffer collateral CA1 pathway, were analyzed for the population spike (PS) amplitude. Pentobarbital had a concentration-dependent, biphasic effect on PS amplitudes, which were increased approximately twofold (P < 0.001) when the slice was exposed to pentobarbital concentrations of 1 and 5 microM and depressed at drug concentrations larger than 10 microM. Pentobarbital (5 microM) did not increase the PS amplitude when stimulation was stopped during exposure to the drug. The enhancement of PS amplitude was suppressed in the presence of 10 microM acetazolamide, a nonselective carbonic anhydrase inhibitor, and when the slice was bathed in CO(2)/HCO(3)(-)-free artificial cerebrospinal fluid. Pretreatment with 1 microM picrotoxin, a GABA(A) receptor antagonist, or 5 microM 2-amino-5 phosphopentanoic acid, a specific NMDA receptor antagonist, also suppressed enhancement of PS amplitude by 5 microM pentobarbital. The results suggest that small concentrations of pentobarbital (1 and 5 microM) enhance synaptic transmission through mechanisms involving GABA(A) and NMDA receptors and the HCO(3)(-) ion. IMPLICATIONS: Enhanced hippocampal synaptic transmission after exposure to subanesthetic concentrations of pentobarbital persists during drug washout. This finding may help to explain why some patients experience excitation and enhanced pain during emergence from anesthesia. PMID- 11726436 TI - Generalized tonic-clonic activity after remifentanil administration. AB - IMPLICATIONS: This is the first report of seizure-like activity in an adult who received remifentanil. This report confirms that opioid administration can be associated with generalized tonic-clonic seizure-like activity. It is suggested that this reaction could be referred to as the "opioid-seizure syndrome." PMID- 11726437 TI - Cardiac arrest from inadvertent overdose of lidocaine hydrochloride through an arterial pressure line flush apparatus. AB - IMPLICATIONS: An overdose of the antiarrhythmic drug, lidocaine hydrochloride, was inadvertently administered to a patient through an arterial pressure monitoring apparatus, resulting in cardiac asystole. This medication error demonstrates to all anesthesia care team members the importance of preventing similar occurrences. PMID- 11726438 TI - The impact of longer-than-average anesthesia times on the billing of academic anesthesiology departments. AB - Academic anesthesiology departments provide clinical services for surgical procedures that have longer-than-average surgical times and correspondingly increased anesthesia times. We examined the financial impact of these longer times in three ways: 1) the estimated loss in revenue if billing were done on a flat-fee system by using industry-averaged anesthesia times; 2) the estimation of incremental operating room (OR) sites necessitated by longer anesthesia times; and 3) the estimated potential gain in billed units if the hours of productivity of current anesthesia time were applied to surgical cases of average duration. Health Care Financing Administration average times per anesthesia procedure code were used as industry averages. Billing data were collected from four academic anesthesiology departments for 1 yr. Each claim billed with ASA units was included except for obstetric anesthesia care. All clinical sites that do not bill with ASA units were excluded. Base units were determined for each anesthesia procedure code. The mean commercial conversion factor (US$45 per ASA unit) for reimbursement was used to estimate the impact in dollar amounts. In all four groups, anesthesia times exceeded the Health Care Financing Administration average. The loss per group in billed ASA units if a flat-fee billing system were used ranged from 18,194 to 31,079 units per group, representing a 5% to 15% decrease (estimated billing decrease of US$818,719 to US$1,398,536 per group). The number of excess OR sites necessitated by longer surgical and anesthesia times ranged from 1.95 to 4.57 OR sites per group. The potential gain in billed units if the hours of productivity of current anesthesia time were applied to surgical cases of average duration was estimated to be from 13,273 to 21,368 ASA units. Longer-than-average anesthesia and surgical times result in extra hours or additional OR sites to be staffed and loss of potential reimbursement for the four academic anesthesiology departments. A flat-fee system would adversely affect academic anesthesiology departments. IMPLICATIONS: We examined the economic impact of longer-than-average anesthesia times on four academic anesthesiology departments in three ways: the estimated loss in revenue under a flat-fee system, the excess operating room sites staffed, and the potential gain in revenue if the surgeries were of average length. These results should be considered both in productivity measurements and strategies for operating room management. PMID- 11726439 TI - Patient threats present an ethical dilemma for the anesthesiologist. AB - IMPLICATIONS: Patients who receive sedation occasionally divulge thoughts that they would not usually express. This report describes a sedated patient who threatened to murder two family members. Immediate consultation with an attorney and psychiatrist is recommended when the anesthesiologist may be required to breach patient confidentiality to warn potential victims. PMID- 11726440 TI - The safety of immediate extubation after abdominal aortic surgery: a prospective, randomized trial. AB - We performed this study to assess the safety of immediate extubation after elective abdominal aortic aneurysm surgery. Consecutive patients were prospectively randomized into two groups after surgery: Group 1 (n = 29) immediate extubation; and Group 2 (n = 21) delayed (at least 4 h) extubation. All patients were assessed by a senior anesthesiologist or intensivist before extubation. The following data were collected: preoperative-demographics, presence of comorbid disease, body mass index, hemoglobin level, heart rate, and blood pressure; intraoperative-duration of surgery and cross-clamping, blood loss, amount of crystalloids, colloids, and blood transfused, temperature at end of procedure, urine output, and complications; and postoperative-time to extubation, scores on the Acute Physiology and Chronic Health Evaluation-II and Therapeutic Intervention Scoring System, total fentanyl dose, and complications. Outcome variables were length of intensive care unit and hospital stay and 28-day mortality. The results showed no significant differences in preoperative or intraoperative variables between the groups, apart from a longer duration of surgery in Group 1 (P = 0.045). Group 2 patients had a significantly higher Therapeutic Intervention Scoring System score (P = 0.04) and required a significantly larger dose of fentanyl (P < 0.001). One patient in Group 2 required reintubation after a cerebrovascular accident. The overall mortality rate was 4% (2 patients in Group 2). There were no significant differences in any of the outcome variables. We conclude that immediate extubation can safely be performed after elective abdominal aortic aneurysm surgery. IMPLICATIONS: In this prospective randomized study, we compared the outcome of patients undergoing elective aortic abdominal surgery who either were extubated immediately after surgery or after 4 h of stabilization in the intensive care unit. No significant differences were found in the length of intensive care unit or hospital stay, or 28-day mortality between the 2 groups. PMID- 11726441 TI - Rapid weaning from mechanical ventilator in acute cervical cord multiple sclerosis lesion after steroids. AB - IMPLICATIONS: We report a patient with multiple sclerosis (MS), who developed neuromuscular respiratory failure requiring ventilation because of a cervical cord relapse. Serial pulmonary function tests documented improvement after steroid treatment. Cervical cord or brainstem relapses should be suspected in MS patients with respiratory failure. Identification and management of this critical condition are discussed. PMID- 11726442 TI - The differential effects of stereoisomers of ropivacaine and bupivacaine on cerebral pial arterioles in dogs. AB - We investigated whether the stereoisomers of ropivacaine and bupivacaine exert differential effects on the cerebral microcirculation. Pentobarbital-anesthetized dogs (n = 16) were prepared for measurement of cerebral pial vessel diameters by using a closed cranial window preparation. We administered three different concentrations (10(-7), 10(-5), and 10(-3) M) of each of three drug solutions [R(+), racemic, and S(-) forms of ropivacaine (n = 8) or bupivacaine (n = 8)] under the window in a randomized manner and measured cerebral pial arteriolar diameters. Various physiologic data were obtained before and after topical application of each test solution. All three forms of ropivacaine constricted cerebral pial arterioles, each in a concentration-dependent manner. The rank order for degree of vasoconstriction was S(-) ropivacaine > racemic ropivacaine > R(+) ropivacaine. In contrast, R(+) and racemic bupivacaine dilated, but S(-) bupivacaine constricted, cerebral pial arterioles, each in a concentration dependent manner. We could find no difference in vascular reactivity to these drugs between large (> or = microm) and small (<100 microm) arterioles. Topical application of these drugs induced no changes in mean blood pressure or heart rate. The observed differences in the microvascular alterations induced by the stereoisomers of ropivacaine and bupivacaine suggest that the vasoactive effects of these drugs on cerebral arterioles could, at least in part, depend on their chirality. IMPLICATIONS: The differential effects of the stereoisomers of ropivacaine and bupivacaine on cerebral pial vessels could, at least in part, depend on their chirality. PMID- 11726443 TI - Paraneoplastic syndromes associated with lung cancer: a unique case of concomitant subacute cerebellar degeneration and Lambert-Eaton myasthenic syndrome. AB - IMPLICATIONS: We report an unusual case in which a patient with paraneoplastic subacute cerebellar degeneration (a brain disorder resulting from antibody production by a tumor located outside the skull) developed Lambert-Eaton Myasthenic Syndrome (antibody-mediated skeletal muscle weakness) that was not apparent until she underwent surgery. Failure to recognize this disease process can cause life-threatening respiratory distress. PMID- 11726444 TI - Neostigmine combined with bupivacaine, clonidine, and sufentanil for spinal labor analgesia. AB - We previously found that spinal clonidine prolongs labor analgesia when combined with spinal bupivacaine and sufentanil. We sought to determine whether the addition of spinal neostigmine to these drugs would further enhance labor analgesia. By use of a combined spinal/epidural technique, 36 patients were randomized to receive a hyperbaric spinal injection of bupivacaine 2.5 mg plus clonidine 50 microg and sufentanil 10 microg with or without neostigmine 10 microg. Pain, maternal hemodynamics, fetal heart rate, nausea, pruritus, sedation, motor block, sensory levels to pinprick, and maternal oxygen saturation were assessed at regularly specified intervals after spinal injection until additional analgesia was requested. The duration of spinal analgesia was similar between groups (215 +/- 60 min in the Control group versus 205 +/- 62 min in the Neostigmine group). Likewise, pain scores, the duration of labor, Apgar scores, and side effects were similar between groups except that patients administered neostigmine experienced significantly more nausea and vomiting (53% vs 7%, P = 0.01). We conclude that spinal neostigmine 10 microg produces severe nausea and does not potentiate the duration of spinal analgesia in laboring women from spinal bupivacaine, clonidine, and sufentanil. IMPLICATIONS: Spinal neostigmine 10 microg as an adjunct to spinal bupivacaine, clonidine, and sufentanil produces severe nausea and fails to potentiate analgesia in laboring women. PMID- 11726445 TI - Early pregnancy does not reduce the C(50) of propofol for loss of consciousness. AB - Requirements for inhaled anesthetics decrease during pregnancy. There are no published data, however, regarding propofol requirements in these patients. Because propofol is often used for induction of general anesthesia when surgery is necessary in early pregnancy, we investigated whether early pregnancy reduces the requirement of propofol for loss of consciousness using a computer-assisted target-controlled infusion (TCI). Propofol was administered using TCI to provide stable concentrations and to allow equilibration between blood and effect-site (central compartment) concentrations. Randomly selected target concentrations of propofol (1.5-4.5 microg/mL) were administered to both pregnant women (n = 36) who were scheduled for pregnancy termination and nonpregnant women (n = 36) who were scheduled for elective orthopedic or otorhinolaryngologic surgery. The median gestation of the pregnant women was 8 wk (range, 6-12 wk). Venous blood samples for analysis of the serum propofol concentration were taken at 3 min and 8 min after equilibration of the propofol concentration. After a 10-min equilibration period of the predetermined propofol blood concentration, a verbal command to open their eyes was given to the patients twice, accompanied by rubbing of their shoulders. Serum propofol concentrations at which 50% of the patients did not respond to verbal commands (C(50) for loss of consciousness) were determined by logistic regression. There was no significant difference in C(50) +/- SE of propofol for loss of consciousness between the Nonpregnant (2.1 +/- 0.2 microg/mL) and Pregnant (2.0 +/- 0.2 microg/mL) groups. These results indicate that early pregnancy does not decrease the concentration of propofol required for loss of consciousness. IMPLICATIONS: The C(50) of propofol for loss of consciousness in early pregnancy did not differ from that in nonpregnant women, indicating that there is no need to decrease the propofol concentration for loss of consciousness when inducing general anesthesia for termination of pregnancy. PMID- 11726446 TI - Hemodynamics during laparoscopic surgery in pregnancy. AB - IMPLICATIONS: During laparoscopic cholecystectomy in four pregnant women, we observed hemodynamic changes similar to those in nonpregnant patients (i.e., decreases in cardiac index together with increases in mean arterial blood pressure and systemic vascular resistance). PMID- 11726447 TI - Chronic blockade of melanocortin receptors alleviates allodynia in rats with neuropathic pain. AB - We investigated the involvement of the spinal cord melanocortin (MC) system in neuropathic pain. Because we recently demonstrated that MC receptor ligands acutely alter nociception in an animal model of neuropathic pain, in this study we tested whether chronic administration was also effective. We hypothesized that chronic blockade of the spinal MC system might decrease sensory abnormalities associated with this condition. The effects of the MC receptor antagonist SHU9119 (0.5 microg/d) and agonist MTII (0.1 microg/d) were evaluated in rats with a chronic constriction injury of the sciatic nerve. Drugs were continuously infused into the cisterna magna. Antinociceptive effects were measured with tests involving temperature (10 degrees C or 47.5 degrees C) or mechanical (von Frey) stimulation. The administration of MTII increased mechanical allodynia, whereas SHU9119 produced a profound cold and mechanical antiallodynia, altering responses to control levels. The antiallodynic effects of SHU9119 were very similar to those produced by the alpha(2)-adrenergic agonist tizanidine (50 microg/d). The effects of SHU9119 and MTII are most likely mediated through the MC4 receptor, because this is the only MC-receptor subtype present in the spinal cord. We conclude that the chronic administration of MC4-receptor antagonists might provide a promising tool in the treatment of neuropathic pain. IMPLICATIONS: In this study we demonstrated that continuous intrathecal infusion of the melanocortin-receptor antagonist SHU9119 reduces cold and mechanical allodynia in rats with a chronic constriction injury of the sciatic nerve, a lesion producing neuropathic pain. PMID- 11726448 TI - Body temperature and diaphoresis disturbances in a patient with arachnoiditis. AB - IMPLICATIONS: Arachnoiditis, produced by different causes, is an inflammation of the sac containing the spinal cord and nerve roots. Patients with this disease have severe low back and leg pain, sweating and low grade fever. This case had aberrant skin temperature and sweating in different parts of the body. PMID- 11726449 TI - Endogenous endothelin and vasopressin support blood pressure during epidural anesthesia in conscious dogs. AB - We studied whether endogenous endothelin, like endogenous vasopressin, helps to maintain blood pressure during high epidural anesthesia when efferent sympathetic drive is diminished. On different days, six awake dogs underwent each of the following five interventions: blockade of vasopressin V(1a) receptors using [d(CH(2))(5)Tyr(Me(2))]AVP, (40 microg/kg) or endothelin receptors using tezosentan (3 mg/kg followed by 3 mg. kg(-1). h(-1)) with or without epidural anesthesia (1% lidocaine, intraindividual dose did not differ between experiments), and epidural saline (n = 5). The effects of endothelin- or vasopressin-receptor blockade were analyzed (means +/- SEM) and compared by an analysis of variance for repeated measures (paired Student's t-test, alpha adjusted, P < 0.05). Vasopressin-receptor blockade decreased blood pressure (10 +/- 2 mm Hg) only in the presence of epidural anesthesia, whereas endothelin receptor blockade reduced blood pressure both in the presence and absence of epidural anesthesia (12 +/- 3 versus 10 +/- 1 mm Hg). During baseline and each intervention, plasma concentrations of vasopressin and big-endothelin were measured and compared by a Wilcoxon's rank sum test; P < 0.05. Vasopressin concentrations increased during epidural anesthesia and after additional endothelin receptor blockade, but big-endothelin concentrations remained unchanged during each intervention. We conclude that vasopressin acts as a reserve system, as it stabilizes blood pressure specifically during epidural anesthesia, whereas the unchanged concentrations of big-endothelin indicate that the endothelin system is not specifically activated to support blood pressure during epidural anesthesia. IMPLICATIONS: We studied in awake dogs whether endogenous endothelin, like endogenous vasopressin, helps to maintain blood pressure during resting conditions and epidural anesthesia. Only vasopressin was specifically activated to support blood pressure during epidural anesthesia, whereas endothelin supported blood pressure to the same extent during epidural anesthesia and during resting conditions. PMID- 11726450 TI - Patient-controlled epidural analgesia after abdominal surgery: ropivacaine versus bupivacaine. AB - In this randomized, double-blinded study we sought to assess the analgesic efficacy of ropivacaine and bupivacaine in combination with sufentanil and the efficacy of ropivacaine alone after major abdominal surgery. Sixty patients undergoing major abdominal surgery received standardized general anesthesia combined with epidural thoracic analgesia. They were allocated to one of three groups: the BS group received postoperative patient-controlled epidural analgesia with 0.125% bupivacaine plus 0.5 microg/mL sufentanil; the RS group received 0.125% ropivacaine plus 0.5 microg/mL sufentanil; and the R group received 0.2% ropivacaine, with the patient-controlled epidural analgesia device set at bolus 2 3 mL and background infusion 3-5 mL/h. Visual analog scale scores were significantly lower during coughing in the BS group compared with the RS and R groups and in the RS group compared with the R group. The BS group required significantly less local anesthetic (milligrams per day) during the first three postoperative days compared with the RS and R groups, and the RS group, significantly less than the R group. No major side effects were noted in any group. We conclude that, after major abdominal surgery, thoracic epidural analgesia was more effective with bupivacaine than with ropivacaine when these two local anesthetics are used in a mixture with sufentanil. Ropivacaine alone was less effective than ropivacaine in combination with sufentanil. IMPLICATIONS: After major abdominal surgery, thoracic epidural analgesia was more effective with 0.125% bupivacaine than with 0.125% ropivacaine when these two local anesthetics were used in a mixture with 0.5 microg/mL sufentanil. Ropivacaine 0.2% alone was less effective than 0.125% ropivacaine combined with sufentanil. PMID- 11726451 TI - A dose response study of clonidine with local anesthetic mixture for peribulbar block: a comparison of three doses. AB - Clonidine prolongs anesthesia and analgesia of local anesthetics in various neural blocks as well as the duration of retrobulbar block. We assessed the dose response relationship of clonidine added to lidocaine in peribulbar block. Sixty patients undergoing cataract surgery were given peribulbar block with 7 mL of 2% lidocaine and hyaluronidase with either saline (Control) or clonidine in 0.5 microg/kg (0.5 Clon), 1.0-microg/kg (1.0 Clon), or 1.5-microg/kg (1.5 Clon) doses. The onset and duration of lid and globe akinesia, globe anesthesia and analgesia, postoperative analgesic requirement, and adverse effects (hypotension, bradycardia, hypoxia, sedation, and dizziness) were recorded. The success rate and onset of block were comparable in all groups. The duration of lid and globe akinesia, globe anesthesia and analgesia was significantly (P < 0.01) prolonged in patients receiving 1.0 and 1.5 microg/kg clonidine as compared with the Control group. Perioperative pain scores and analgesic requirement were significantly less in these groups. 0.5 microg/kg clonidine did not increase the duration of anesthesia and analgesia significantly. Hypotension and dizziness were observed more in patients receiving 1.5 microg/kg clonidine as compared with other groups. We conclude that 1.0 microg/kg clonidine with a mixture of lidocaine (2%) significantly prolonged the duration of anesthesia and analgesia after peribulbar block with limited side effects. IMPLICATIONS: We studied the effect of the addition of 0.5, 1.0 and 1.5 microg/kg clonidine to a lidocaine hyaluronidase mixture on the onset and duration of peribulbar block and perioperative analgesia. A dose of 1.0 microg/kg produced a significant increase in duration of anesthesia and analgesia with minimal side effects. PMID- 11726452 TI - The comparative electrophysiologic and hemodynamic effects of a large dose of ropivacaine and bupivacaine in anesthetized and ventilated piglets. AB - Ropivacaine is less potent and less toxic than bupivacaine. We administered these two local anesthetics in a cardiac electrophysiologic model of sodium thiopental anesthetized and ventilated piglets. After assessing the stability of the model, bupivacaine (4 mg/kg) and ropivacaine (6 mg/kg) were given IV in two groups (n = 7) of piglets. No alteration in biological variables was reported throughout the study. Bupivacaine and ropivacaine similarly decreased mean aortic pressure from 99 +/- 22 to 49 +/- 31 mm Hg and from 87 +/- 17 to 58 +/- 28 mm Hg, respectively, and decreased the peak of the first derivative of left ventricular pressure from 1979 +/- 95 to 689 +/- 482 mm Hg/s and from 1963 +/- 92 to 744 +/- 403 mm Hg/s, respectively. Left ventricular end-diastolic pressure was similarly increased from 6 +/- 5 to 9 +/- 5 mm Hg and from 6 +/- 4 to 12 +/- 4 mm Hg, respectively. Bupivacaine and ropivacaine similarly lengthened the cardiac cycle length (R-R; from 479 +/- 139 to 706 +/- 228 ms and from 451 +/- 87 to 666 +/- 194 ms, respectively), atria His (from 71 +/- 15 to 113 +/- 53 ms and from 64 +/- 6 to 86 +/- 10 ms, respectively), and QTc (QTc = QT x R-R(-0.5), Bazett formula; from 380 +/- 71 to 502 +/- 86 ms and from 361 +/- 33 to 440 +/- 56 ms, respectively) intervals. Bupivacaine altered to a greater extent the PQ (the onset of the P wave to the Q wave of the QRS complex) (from 97 +/- 20 to 211 +/- 60 ms versus from 91 +/- 8 to 145 +/- 38 ms, P < 0.05), QRS (from 58 +/- 3 to 149 +/- 34 ms versus from 60 +/- 5 to 101 +/- 17 ms, P < 0.05), and His ventricle interval (from 25 +/- 4 to 105 +/- 30 ms vs from 25 +/- 4 to 60 +/- 30 ms, P < 0.05) than ropivacaine. A 6 mg/kg ropivacaine dose induced similar hemodynamic alterations as 4 mg/kg bupivacaine. However, bupivacaine altered the variables of ventricular conduction (QRS and His ventricle) to a greater extent. IMPLICATIONS: A 6 mg/kg ropivacaine dose induced similar hemodynamic alterations as 4 mg/kg bupivacaine. However, bupivacaine altered the variables of ventricular conduction (QRS and His ventricle) to a greater extent. PMID- 11726453 TI - Regional anesthesia does not increase the risk of postoperative neuropathy in patients undergoing ulnar nerve transposition. AB - The use of regional anesthetic techniques in patients with preexisting neuropathies has been widely debated. The possibility of needle- or catheter induced trauma, local anesthetic toxicity, or neural ischemia during regional blockade may place patients with underlying mechanical, ischemic, or metabolic neurologic derangements at increased risk of progressive neural injury. We evaluated the safety of regional versus general anesthesia in patients with a preexisting ulnar neuropathy undergoing ulnar nerve transposition. All patients (n = 360) who underwent ulnar nerve transposition at the Mayo Clinic from 1985 to 1999 were retrospectively studied. A general anesthetic was performed in 260 (72%) patients. The remaining 100 (28%) patients received an axillary block, including 64 patients in whom an ulnar paresthesia or nerve stimulator motor response was elicited at the time of block placement. Patient characteristics, the severity of preoperative ulnar nerve dysfunction, and surgical variables were similar between groups. Anesthetic technique did not affect neurologic outcome (new or worsening pain, paresthesias, numbness, or motor weakness) immediately after surgery or at 2 or 6 wk after surgery. All six patients in the Axillary Block group who reported new or worsening neurologic symptoms after surgery had received bupivacaine in combination with either an ulnar paresthesia or motor response. By using logistic regression, bupivacaine was identified as an independent risk factor for worsening of ulnar nerve function compared with other local anesthetics. We conclude that axillary blockade is a suitable anesthetic technique for this procedure. IMPLICATIONS: The use of regional anesthetic techniques in patients with preexisting neuropathies has been widely debated. Theoretical concerns include the risk of progressive nerve damage from direct needle trauma or local anesthetic toxicity. This investigation, however, supports the safety of axillary blockade in patients with preexisting ulnar neuropathy undergoing ulnar nerve transposition. PMID- 11726455 TI - The use of a "reverse" axis (axillary-interscalene) block in a patient presenting with fractures of the left shoulder and elbow. AB - IMPLICATIONS: A patient presented for surgery to repair a fractured left shoulder and elbow and requested regional anesthesia. Most upper extremity operations require a single brachial plexus nerve block. The position of the two fractures however required the use of two separate approaches, an interscalene and an axillary approach. PMID- 11726454 TI - The efficacy of simulated intravascular test dose in sedated patients. AB - Sedation usually decreases the reliability of subjectively detecting an intravascular test dose, but the efficacies of objective hemodynamic and T-wave criteria remain undetermined. Sixty healthy patients were randomly assigned to receive IV midazolam in 1-mg increments until they were lightly sedated, fentanyl 2 microg/kg followed by incremental midazolam until they were similarly sedated, or no sedative (n = 20 each). Then, normal saline 3 mL was administered IV, followed 4 min later by 1.5% lidocaine 3 mL plus epinephrine 15 microg (1:200,000) in all subjects. Heart rate (HR), systolic blood pressure (SBP) measured by a radial arterial catheter, and lead II of the electrocardiogram were continuously recorded for 4 min after the saline and test dose injections. An IV test dose produced significant increases in HR and SBP and decreases in T-wave amplitude in all subjects. However, the mean maximum increase in HR in patients sedated with midazolam plus fentanyl (31 +/- 14 bpm [mean +/- SD]) was significantly less than in those administered midazolam alone or no sedative (42 +/- 12 and 44 +/- 10 bpm, respectively; P < 0.05). A sensitivity of 100% was obtained on the basis of the traditional HR criterion (positive if > or =20 bpm increase) in patients sedated with midazolam or no sedative, but it was 70% in those with midazolam plus fentanyl (P < 0.05 versus the other two groups). Irrespective of the treatment, sensitivities and specificities of 100% were obtained according to the SBP (positive if > or =15 mm Hg increase) and T-wave (positive if > or =25% decrease in amplitude) criteria. An increase in SBP and a decrease in T-wave amplitude are more reliable than an HR response for detecting accidental intravascular injection of the epinephrine-containing test dose in subjects sedated with midazolam and fentanyl. IMPLICATIONS: To determine whether an epidural catheter is in a blood vessel, an epidural test dose containing 15 microg epinephrine is used. We found that an increase in systolic blood pressure and a decrease in T-wave amplitude seem to be more reliable than a heart rate change for detecting inadvertent intravascular injection of an epinephrine containing test dose in patients sedated with midazolam and fentanyl. PMID- 11726456 TI - Gastric residue is not more copious in obese patients. AB - IMPLICATIONS: Previous studies have shown that obese patients have a larger volume of gastric content than lean patients do. However, methodological limitations call into question the validity of these findings. We have reexamined this issue and found identical gastric content volumes in fasting obese and lean subjects after an 8-h fast. PMID- 11726457 TI - A multimodal postoperative rehabilitation program in critically ill diabetic patients. PMID- 11726458 TI - Play stickers: pediatric mask induction made easy. PMID- 11726459 TI - Difficult intubation and difficult airway. PMID- 11726460 TI - Selective bronchial blockade in small infants. PMID- 11726461 TI - Pain therapy with interscalene local anesthetic. PMID- 11726462 TI - About the origin of the PaCO(2)/ETCO(2) ratio. PMID- 11726463 TI - What to prefer during jet ventilation for endolaryngeal procedures, risk of barotrauma or of aspiration of contaminating material? PMID- 11726464 TI - Acetaminophen analgesia in infants. PMID- 11726465 TI - Analgesic efficacy of tramadol if coadministered with ondansetron. PMID- 11726466 TI - TEE diagnosis of mechanical AVR dysfunction associated with biological glue. PMID- 11726467 TI - Cerebral oxygenation and cognitive decline. PMID- 11726468 TI - Raiders of the lost dream. PMID- 11726469 TI - Warm up. PMID- 11726470 TI - When to retire after concussion? PMID- 11726471 TI - A systematic review of interventions to prevent lower limb soft tissue running injuries. AB - OBJECTIVES: To assess the available evidence for preventive strategies for lower limb soft tissue injuries caused by running. METHODS: An electronic database search was conducted using The Cochrane Musculoskeletal Injuries Group Specialised Register, The Cochrane Controlled Trials Register, Medline, Embase, Sport Discus, Heracles, Atlantes, Biosis, Cinahl, Scisearch, Current Contents, Index To Theses and Dissertation Abstracts. Any randomised or quasi-randomised trials evaluating interventions to prevent running injuries to lower limb soft tissue were included. The eligibility of trials for inclusion and the quality of the trials were independently assessed by two reviewers. RESULTS: Exposure to a high training load (duration, frequency, or running distance) increases the risk of injury, and thus modification of the training schedule can reduce the incidence of injury. The effectiveness of stretching exercises and of insoles in the prevention of lower extremity soft tissue injuries caused by running is not known. Wearing a knee brace with a patellar support ring may be effective in the prevention of anterior knee pain caused by running. CONCLUSIONS: This review provides evidence for the effectiveness of the modification of training schedules in reducing lower limb soft tissue running injuries. More studies are required to quantify the optimal training loads and to confirm that knee braces can prevent knee pain. It is important to note that the studies included in this review had few female participants therefore the results may not be generalisable. PMID- 11726472 TI - Effects of rapid weight loss on mood and performance among amateur boxers. AB - AIMS: To examine the effects of rapid weight loss on mood and performance among amateur boxers. METHODS: Participants were 16 experienced amateur boxers. In stage 1, structured interviews were used to assess the type of strategies that boxers used to reduce weight and the value of performing at their desired weight in terms of performance. In stage 2, boxers completed a 4 x 2 minute (1 minute recovery) circuit training session. Boxers completed the circuit training session on three different occasions with a week between each. The first test was used to familiarise the boxers with the circuit training task; the second and third tasks were at their training weight and championship weight, respectively. Participants were given one week to reduce their body weight to their championship weight using their preferred weight making strategies; boxers reduced their body weight by an average of 5.16% of body weight. RESULTS: Boxers typically lost weight by restricting fluid and food intake in the week leading to competition. Repeated measures multivariate analysis of variance results indicated that rapid weight loss among boxers was associated with poor performance, increased anger, fatigue, and tension, and reduced vigour. CONCLUSIONS: Strategies used to make weight by boxers are associated with poor performance and a negative mood profile. PMID- 11726473 TI - Physiological and metabolic characteristics of elite tug of war athletes. AB - OBJECTIVE: To determine the aerobic power (VO(2)MAX), body composition, strength, muscular power, flexibility, and biochemical profile of an elite international squad of tug of war athletes. METHODS: Sixteen male competitors (mean (SEM) age 34 (2) years) were evaluated in a laboratory. For comparative purposes, data were analysed relative to normative data for our centre and to a group of 20 rugby forwards from the Irish international squad. RESULTS: The tug of war participants were lighter (83.6 (3.0) v 104.4 (1.8) kg, p<0.0001) and had less lean body mass (69.4 (2.1) v 86.2 (1.2) kg) than the rugby players and had lower than normal body fat (16.7 (0.9)%); all values are mean (SEM). Aerobic power measured during a treadmill test was 55.8 (1.6) ml/kg/min for the tug of war participants compared with 51.1 (1.4) ml/kg/min for the rugby forwards (p<0.03). A composite measure of strength derived from (sum of dominant and non-dominant grip strength and back strength)/lean body mass yielded a strength/mass ratio that was 32% greater (p<0.0001) for the tug of war group than the rugby group. Dynamic leg power was lower for the tug of war group than the rugby forwards (4659.8 (151.6) v 6198.2 (105) W respectively; p<0.0001). Leg flexibility was 25.4 (2.0) cm for the tug of war group. Back flexibility was 28.6 (1.4) cm which was lower (p<0.02) than the rugby forwards 34.2 (1.5) cm. Whereas blood chemistry and haematology were normal, packed cell volume, haemoglobin concentration, and erythrocyte volume were lower in the tug of war group than in the rugby players (p<0.05). All three haematological measures correlated with muscle mass (packed cell volume, r(2) = 0.37, p<0.0001; haemoglobin concentration, r(2) = 0.13, p<0.05; erythrocyte volume, r(2) = 0.21, p<0.01). CONCLUSIONS: The data indicate that international level tug of war participants have excellent strength and above average endurance relative to body size, but have relatively low explosive leg power and back flexibility. The data provide reference standards for the sport and may be useful for monitoring and evaluating current and future participants. PMID- 11726476 TI - Distal radial fractures in young goalkeepers: a case for an appropriately sized soccer ball. AB - OBJECTIVES: To assess the rate of wrist fractures in young goalkeepers sustained by the specific mechanism of "saving the ball" and the potential influence of ball size and environmental conditions. METHODS: A prospective, clinic based study in one institution over a 17 month period. Patients were identified by specific questioning. Information on play circumstances and subsequent clinical progress was documented. RESULTS: Twenty nine fractures of the distal radius were identified in young goalkeepers (age range 6-15 years) as a direct result of saving the ball. Most were managed simply in a plaster cast. Three patients required minor surgical interventions, and all fractures went on to unite without significant complications. Where ball size was known, 12 of the 15 fractures in children aged 11 years or less occurred as the result of impact with an adult sized ball compared with three when a junior ball was involved. This is statistically significant (p = 0.039). In the 10 children aged 12-15 years, only one fracture involved a junior ball; this is also statistically significant (p = 0.027). Six of the injuries (21%) occurred when the ball was kicked by an adult. Injuries occurred in both organised and informal games throughout the year. CONCLUSIONS: This specific mechanism of injury has not been widely acknowledged nor has the potential influence of ball size as a causative factor been examined. Recommendations for an appropriately sized soccer ball for young players exist but are not in universal use. Increased awareness of this particular injury mechanism is required. PMID- 11726474 TI - Propensity for osteoarthritis and lower limb joint pain in retired professional soccer players. AB - OBJECTIVE: To quantify the prevalence of osteoarthritis and the severity of pain in the lower limb joints of players retired from English professional soccer. METHOD: An anonymous self administered questionnaire was distributed to 500 former players registered with the English Professional Footballers' Association. The questionnaire was designed to gather information on personal details, physical activity loading patterns, history of lower limb joint injury, and current medical condition of the lower limb joints. RESULTS: Of 500 questionnaires distributed, 185 (37%) were returned. Nearly half of the respondents (79: 47%) retired because of injury; 42% (33) were acute injuries and 58% (46) chronic injuries. Most of the acute injuries that led to early retirement were of the knee (15: 46%), followed by the ankle (7: 21%) and lower back (5: 15%). Most of the chronic injuries that led to early retirement were also of the knee (17: 37%), followed by the lower back (10: 22%) and the hip (4: 9%). Of all respondents, 32% (59) had been medically diagnosed with osteoarthritis in at least one of the lower limb joints. More respondents had been diagnosed with osteoarthritis in the knee joints than either the ankle or the hip joints. Significantly (p<0.001) more respondents reported pain in one lower extremity joint during one or more daily activities than those who did not (joint pain: 137, 80%; no joint pain: 35, 20%). CONCLUSION: The risk for professional soccer players of osteoarthritis in at least one of the lower extremity joints is very high and significantly greater than for the general population. The results support the suggestion that professional soccer players should be provided with health surveillance during their playing career. PMID- 11726477 TI - Influence of preseason training, fitness, and existing injury on subsequent rugby injury. AB - OBJECTIVES: To examine the influence of preseason fitness, existing injury, and preseason rugby training on subsequent injury. METHODS: Players were eligible for the survey if they were a member of a Scottish Rugby Union (SRU) affiliated rugby club in the Border Reivers District of the SRU during the 1997-1998 season. A total of 803 (84%) players from 22 (88%) participating clubs provided details of rugby training, injuries sustained, and physical activity undertaken during the 16 week summer period (26 April to 16 August 1997) and their perceived fitness before the start of the season. Observers at participating clubs reported all injury episodes occurring to club players throughout the 1997-1998 season. RESULTS: One fifth of players did not attend any rugby training during the 16 week summer period; the remainder attended a median of 14 sessions. Throughout the 1997-1998 season, 675 injury episodes occurred to 423 (53%) players during training or in matches. After adjustment for whether players held a professional contract or were amateurs, Cox regression showed a 3.9% relative increase (95% confidence interval (CI) 1.9 to 5.9%) in the risk of injury over the season for each additional preseason training week attended, and a 61% relative increase (95% CI 32 to 97%) for those players who had been injured or were carrying an injury at the end of the previous season. CONCLUSIONS: Injury risk is more likely to be related to rugby training (type of activities undertaken in rugby training, or personalities and characteristics of players undertaking training more frequently) than to overall player fitness. Players who were injured at the end of the previous season were more likely to be injured in the following season. This may be because they do not allow previous injuries to heal sufficiently before returning to the game, or the intensity of their participation may increase their risk of injury. PMID- 11726478 TI - Evidence for the aetiology of injuries in Australian football. AB - OBJECTIVES: To determine in Australian football (a) the influence of ground hardness and playing grade (level) on game speed and structure, and (b) player movement patterns throughout the game and across levels. METHODS: The design consisted of several studies. Seventeen games played on grounds of different hardness in 2000 were used to determine game speed and structure. Four first grade and four second grade grand final games (1994, 1996, 1997, 1999) were used to determine the game speed and structure on the same ground but at different levels. Fifty one players (44 first grade and seven second grade) were used to measure movement patterns within games and across levels during the 2000 season. RESULTS: There was a significant relation between ground hardness and game speed, which could lead to higher injury rates when the ground is harder. There was a 6.7% difference in game speed between the first and second grade levels reflecting differences in injury incidence. The first grade games were also characterised by a greater number of shorter, high intensity play periods and longer stop periods than the second grade games. Midfield players in the first grade games covered about 24% greater distance than their second grade counterparts, and there was a significant difference in their playing speeds. CONCLUSIONS: Over the past 40 years, the game speed in the top level of Australian football has approximately doubled. Over the same time, the number of collisions and the estimated injury incidence have also doubled. This study provides additional support to the suggestion that these variables are strongly linked. Factors such as ground hardness, playing level, and time during the game influence game speed and are therefore important in injury development in Australian football. PMID- 11726480 TI - Hormone levels of world class cyclists during the Tour of Spain stage race. AB - OBJECTIVES: To evaluate the hormonal response to strenuous endurance exercise performed by elite athletes. METHODS: Nine professional cyclists (mean (SD) age 28 (1) years; mean (SD) VO(2)MAX 75.3 (2.3) ml/kg/min) who participated in a three week tour race (Vuelta a Espana 1999) were selected as subjects. Morning urinary levels of 6-sulphatoxymelatonin (aMT6s) and morning serum levels of testosterone, follicle stimulating (FSH), luteinising hormone (LH), and cortisol were measured in each subject at t(0) (before the competition), t(1) (end of first week), t(2) (end of second week), and t(3) (end of third week). Urine samples of aMT6s were also evaluated in the evening at t(0), t(1), t(2), and t(3). RESULTS: Mean urinary aMT6s levels had increased significantly (p<0.01) during the day after each stage (1091 (33) v 683 (68) ng/ml at t(1); 955 (19) v 473 (53) ng/ml at t(2); 647 (61) v 337 (47) ng/ml at t(3)). Both morning and evening aMT6s levels decreased significantly during the study. A similar pattern was observed for morning serum levels of cortisol and testosterone. CONCLUSIONS: The results suggest that the basal activity of the pineal gland, adrenal glands, and testis may be decreased after consecutive days of intense, long term exercise. PMID- 11726481 TI - A reduction in training volume and intensity for 21 days does not impair performance in cyclists. AB - OBJECTIVES: (a) To investigate the effects of reduced training on physical condition and performance in well trained cyclists; (b) to study whether an intermittent exercise programme would maintain physiological training adaptations more effectively than a continuous exercise programme during a period of reduced training. METHODS: Twelve male cyclists participated in a 21 day training programme and were divided into two training groups. One group (age 25.3 (7) years; weight 73.3 (5.7) kg; VO(2)MAX 58.6 (4.5) ml/kg/min; means (SD)) underwent a continuous endurance exercise training programme (CT) whereas the second group (age 22.8 (3.5) years; weight 74.1 (7.0) kg; VO(2)MAX 59.7 (6.7) ml/kg/min) followed an intermittent endurance exercise training programme (IT). During this reduced training period, both groups trained for two hours a day, three days a week. RESULTS: Neither group showed changes in maximal workload (WMAX) (4.6 (0.5) v 4.8 (0.5) W/kg and 4.6 (0.5) v 4.7 (0.6) W/kg for the CT and IT group respectively) and VO(2)MAX (58.6 (4.5) v 60.1 (5.8) ml/kg/min and 59.7 (6.7) v 58.8 (7.5) ml/kg/min for the CT and IT group respectively). During the submaximal steady state exercise test, substrate use and heart rate remained unchanged after reduced training. CONCLUSIONS: These results indicate that well trained cyclists who reduce training intensity and volume for 21 days can maintain physiological adaptations, as measured during submaximal and maximal exercise. An intermittent training regimen has no advantage over a continuous training regimen during a detraining period. PMID- 11726483 TI - Clinical risk factors for hamstring muscle strain injury: a prospective study with correlation of injury by magnetic resonance imaging. AB - OBJECTIVE: To prospectively establish risk factors for hamstring muscle strain injury using magnetic resonance imaging (MRI) to define the diagnosis of posterior thigh injury. METHOD: In a prospective cohort study using two elite Australian Rules football clubs, the anthropometric characteristics and past clinical history of 114 athletes were recorded. Players were followed throughout the subsequent season, with posterior thigh injuries being documented. Hamstring intramuscular hyperintensity on T2 weighted MRI was required to meet our criteria for a definite hamstring injury. Statistical associations were sought between anthropometric and previous clinical characteristics and hamstring muscle injury. RESULTS: MRI in 32 players showed either hamstring injury (n = 26) or normal scans (n = 6). An association existed between a hamstring injury and each of the following: increasing age, being aboriginal, past history of an injury to the posterior thigh or knee or osteitis pubis (all p<0.05). These factors were still significant when players with a past history of posterior thigh injury (n = 26) were excluded. Previous back injury was associated with a posterior thigh injury that looked normal on MRI scan, but not with an MRI detected hamstring injury. CONCLUSIONS: Hamstring injuries are common in Australian football, and previous posterior thigh injury is a significant risk factor. Other factors, such as increasing age, being of aboriginal descent, or having a past history of knee injury or osteitis pubis, increase the risk of hamstring strain independently of previous posterior thigh injury. However, as the numbers in this study are small, further research is needed before definitive statements can be made. PMID- 11726485 TI - Non-invasive quantitative assessment of oxidative metabolism in quadriceps muscles by near infrared spectroscopy. AB - BACKGROUND: Near infrared spectroscopy can be used in non-invasive monitoring of changes in skeletal muscle oxygenation in exercising subjects. OBJECTIVE: To evaluate whether this method can be used to assess metabolic capacity of muscles. Two distinctive variables abstracted from a curve of changes in muscle oxygenation were assessed. METHODS: Exercise on a cycle ergometer was performed by 18 elite male athletes and eight healthy young men. A measuring probe was placed on the skin of the quadriceps muscle to measure reflected light at two wavelengths (760 and 850 nm), so that the relative index of muscle oxygenation could be calculated. Exercise intensity was increased from 50 W in 50 W increments until the subject was exhausted. During exercise, changes in muscle oxygenation and blood lactate concentration were recorded. The following two variables for assessment of muscle oxygenation were then abstracted and analysed by plotting curves of changes in muscle oxygenation: the rate of recovery of muscle oxygen saturation (R(R)) and the relative value of the effective decrease in muscle oxygenation (D(eff)). RESULTS: Data analysis showed a correlation between muscle oxygenation and blood lactate concentration at the various exercise intensities and verified the feasibility of the experiment. Data for the athletes were compared with those for the controls using the Aspin-Welch test of significance; t = 2.3 and 2.86 for R(R) and D(eff) respectively. There were significant differences (p = 0.05) between the athletes and the control group with respect to these two variables. CONCLUSION: R(R) and D(eff) may be distinctive variables that can be used to characterise muscle oxidative metabolism during human body movement. PMID- 11726486 TI - Validation of Ottawa ankle rules protocol in Greek athletes: study in the emergency departments of a district general hospital and a sports injuries clinic. AB - OBJECTIVE: To validate the Ottawa ankle rules protocol for predicting ankle and midfoot fractures in Greek athletes. METHOD: A prospective survey in the emergency departments of a district general hospital and a sports injury clinic in Greece over nine months. A clinical evaluation was made of 122 patients with acute ankle and/or midfoot injury, and then radiographs were taken. RESULTS: Nine ankle and eight midfoot fractures were detected. The sensitivity of the Ottawa ankle rules protocol in predicting fractures in both the malleolar and midfoot zones was 100%. The negative predictive value for each of these areas was also 1.0. Specificity was estimated to be 0.3 for ankle fractures and 0.4 for midfoot fractures. Positive predictive values were 0.16 and 0.28 respectively. A possible reduction of up to 28.7% was found in the need for radiography. CONCLUSIONS: Use of the Ottawa ankle rules protocol in evaluating injured Greek athletes resulted in 100% sensitivity when performed by orthopaedic residents or sports medicine doctors, and had the potential to reduce the use of radiography. PMID- 11726488 TI - Stress fracture of the clavicle in a young female gymnast. AB - Stress fractures of the clavicle are rare. This is the report of one such fracture in a 10 year old female gymnast, who presented with a six week history of medial clavicular pain. Radiographs and a computed tomography scan showed an undisplaced fracture through the medial third of the clavicle extending inferiorly to the rhomboid fossa in the inferior aspect of the clavicle. Pathological fracture was excluded by magnetic resonance imaging. The patient was treated conservatively and was able to return to full training eight weeks later. PMID- 11726489 TI - Proportions of activities in a climbing gym. PMID- 11726490 TI - Impact of professionalism on injuries in rugby union. PMID- 11726491 TI - Sports doctors' resuscitation skills. PMID- 11726492 TI - Structures of F420H2:NADP+ oxidoreductase with and without its substrates bound. AB - Cofactor F420 is a 5'-deazaflavin derivative first discovered in methanogenic archaea but later found also to be present in some bacteria. As a coenzyme, it is involved in hydride transfer reactions and as a prosthetic group in the DNA photolyase reaction. We report here for the first time on the crystal structure of an F420-dependent oxidoreductase bound with F420. The structure of F420H2:NADP+ oxidoreductase resolved to 1.65 A contains two domains: an N terminal domain characteristic of a dinucleotide-binding Rossmann fold and a smaller C-terminal domain. The nicotinamide and the deazaflavin part of the two coenzymes are bound in the cleft between the domains such that the Si-faces of both face each other at a distance of 3.1 A, which is optimal for hydride transfer. Comparison of the structures bound with and without substrates reveals that of the two substrates NADP has to bind first, the binding being associated with an induced fit. PMID- 11726493 TI - Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases. AB - Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease processing enzyme. The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme. PMID- 11726494 TI - V-shaped structure of glutamyl-tRNA reductase, the first enzyme of tRNA-dependent tetrapyrrole biosynthesis. AB - Processes vital to life such as respiration and photosynthesis critically depend on the availability of tetrapyrroles including hemes and chlorophylls. tRNA dependent catalysis generally is associated with protein biosynthesis. An exception is the reduction of glutamyl-tRNA to glutamate-1-semialdehyde by the enzyme glutamyl-tRNA reductase. This reaction is the indispensable initiating step of tetrapyrrole biosynthesis in plants and most prokaryotes. The crystal structure of glutamyl-tRNA reductase from the archaeon Methanopyrus kandleri in complex with the substrate-like inhibitor glutamycin at 1.9 A resolution reveals an extended yet planar V-shaped dimer. The well defined interactions of the inhibitor with the active site support a thioester-mediated reduction process. Modeling the glutamyl-tRNA onto each monomer reveals an extensive protein-tRNA interface. We furthermore propose a model whereby the large void of glutamyl-tRNA reductase is occupied by glutamate-1-semialdehyde-1,2-mutase, the subsequent enzyme of this pathway, allowing for the efficient synthesis of 5-aminolevulinic acid, the common precursor of all tetrapyrroles. PMID- 11726495 TI - Specific roles of protein-phospholipid interactions in the yeast cytochrome bc1 complex structure. AB - Biochemical data have shown that specific, tightly bound phospholipids are essential for activity of the cytochrome bc1 complex (QCR), an integral membrane protein of the respiratory chain. However, the structure and function of such phospholipids are not yet known. Here we describe five phospholipid molecules and one detergent molecule in the X-ray structure of yeast QCR at 2.3 A resolution. Their individual binding sites suggest specific roles in facilitating structural and functional integrity of the enzyme. Interestingly, a phosphatidylinositol molecule is bound in an unusual interhelical position near the flexible linker region of the Rieske iron-sulfur protein. Two possible proton uptake pathways at the ubiquinone reduction site have been identified: the E/R and the CL/K pathway. Remarkably, cardiolipin is positioned at the entrance to the latter. We propose that cardiolipin ensures structural integrity of the proton-conducting protein environment and takes part directly in proton uptake. Site-directed mutagenesis of ligating residues confirmed the importance of the phosphatidylinositol- and cardiolipin-binding sites. PMID- 11726496 TI - Crystal structure of the fission yeast mitochondrial Holliday junction resolvase Ydc2. AB - Resolution of Holliday junctions into separate DNA duplexes requires enzymatic cleavage of an equivalent strand from each contributing duplex at or close to the point of strand exchange. Diverse Holliday junction-resolving enzymes have been identified in bacteria, bacteriophages, archaea and pox viruses, but the only eukaryotic examples identified so far are those from fungal mitochondria. We have now determined the crystal structure of Ydc2 (also known as SpCce1), a Holliday junction resolvase from the fission yeast Schizosaccharomyces pombe that is involved in the maintenance of mitochondrial DNA. This first structure of a eukaryotic Holliday junction resolvase confirms a distant evolutionary relationship to the bacterial RuvC family, but reveals structural features which are unique to the eukaryotic enzymes. Detailed analysis of the dimeric structure suggests mechanisms for junction isomerization and communication between the two active sites, and together with site-directed mutagenesis identifies residues involved in catalysis. PMID- 11726497 TI - Crystal structure of the CENP-B protein-DNA complex: the DNA-binding domains of CENP-B induce kinks in the CENP-B box DNA. AB - The human centromere protein B (CENP-B), one of the centromere components, specifically binds a 17 bp sequence (the CENP-B box), which appears in every other alpha-satellite repeat. In the present study, the crystal structure of the complex of the DNA-binding region (129 residues) of CENP-B and the CENP-B box DNA has been determined at 2.5 A resolution. The DNA-binding region forms two helix turn-helix domains, which are bound to adjacent major grooves of the DNA. The DNA is kinked at the two recognition helix contact sites, and the DNA region between the kinks is straight. Among the major groove protein-bound DNAs, this 'kink straight-kink' bend contrasts with ordinary 'round bends' (gradual bending between two protein contact sites). The larger kink (43 degrees ) is induced by a novel mechanism, 'phosphate bridging by an arginine-rich helix': the recognition helix with an arginine cluster is inserted perpendicularly into the major groove and bridges the groove through direct interactions with the phosphate groups. The overall bending angle is 59 degrees, which may be important for the centromere specific chromatin structure. PMID- 11726498 TI - The structure of the negative transcriptional regulator NmrA reveals a structural superfamily which includes the short-chain dehydrogenase/reductases. AB - NmrA is a negative transcriptional regulator involved in the post-translational modulation of the GATA-type transcription factor AreA, forming part of a system controlling nitrogen metabolite repression in various fungi. X-ray structures of two NmrA crystal forms, both to 1.8 A resolution, show NmrA consists of two domains, including a Rossmann fold. NmrA shows an unexpected similarity to the short-chain dehydrogenase/reductase (SDR) family, with the closest relationship to UDP-galactose 4-epimerase. We show that NAD binds to NmrA, a previously unreported nucleotide binding property for this protein. NmrA is unlikely to be an active dehydrogenase, however, as the conserved catalytic tyrosine in SDRs is absent in NmrA, and thus the nucleotide binding to NmrA could have a regulatory function. Our results suggest that other transcription factors possess the SDR fold with functions including RNA binding. The SDR fold appears to have been adapted for other roles including non-enzymatic control functions such as transcriptional regulation and is likely to be more widespread than previously recognized. PMID- 11726499 TI - Apoptosis-associated release of Smac/DIABLO from mitochondria requires active caspases and is blocked by Bcl-2. AB - Smac/DIABLO is a mitochondrial protein that potentiates some forms of apoptosis, possibly by neutralizing one or more members of the IAP family of apoptosis inhibitory proteins. Smac has been shown to exit mitochondria and enter the cytosol during apoptosis triggered by UV- or gamma-irradiation. Here, we report that Smac/DIABLO export from mitochondria into the cytosol is provoked by cytotoxic drugs and DNA damage, as well as by ligation of the CD95 death receptor. Mitochondrial efflux of Smac/DIABLO, in response to a variety of pro apoptotic agents, was profoundly inhibited in Bcl-2-overexpressing cells. Thus, in addition to modulating apoptosis-associated mitochondrial cytochrome c release, Bcl-2 also regulates Smac release, suggesting that both molecules may escape via the same route. However, whereas cell stress-associated mitochondrial cytochrome c release was largely caspase independent, release of Smac/DIABLO in response to the same stimuli was blocked by a broad-spectrum caspase inhibitor. This suggests that apoptosis-associated cytochrome c and Smac/DIABLO release from mitochondria do not occur via the same mechanism. Rather, Smac/DIABLO efflux from mitochondria is a caspase-catalysed event that occurs downstream of cytochrome c release. PMID- 11726500 TI - Wide spectrum of tumors in knock-in mice carrying a Cdk4 protein insensitive to INK4 inhibitors. AB - We have introduced a point mutation in the first coding exon of the locus encoding the cyclin-dependent kinase 4 (Cdk4) by homologous recombination in embryonic stem cells. This mutation (replacement of Arg24 by Cys) was first found in patients with hereditary melanoma and renders Cdk4 insensitive to INK4 inhibitors. Here, we report that primary embryonic fibroblasts expressing the mutant Cdk4R24C kinase are immortal and susceptible to transformation by Ras oncogenes. Moreover, homozygous Cdk4(R24C/R24C) mutant mice develop multiple tumors with almost complete penetrance. The most common neoplasia (endocrine tumors and hemangiosarcomas) are similar to those found in pRb(+/-) and p53(-/-) mice. This Cdk4 mutation cooperates with p53 and p27(Kip1) deficiencies in decreasing tumor latency and favoring development of specific tumor types. These results provide experimental evidence for a central role of Cdk4 regulation in cancer and provide a valuable model for testing the potential anti-tumor effect of Cdk4 inhibitors in vivo. PMID- 11726501 TI - Bub3 interaction with Mad2, Mad3 and Cdc20 is mediated by WD40 repeats and does not require intact kinetochores. AB - The kinetochore checkpoint pathway, involving the Mad1, Mad2, Mad3, Bub1, Bub3 and Mps1 proteins, prevents anaphase entry and mitotic exit by inhibiting the anaphase promoting complex activator Cdc20 in response to monopolar attachment of sister kinetochores to spindle fibres. We show here that Cdc20, which had previously been shown to interact physically with Mad2 and Mad3, associates also with Bub3 and association is up-regulated upon checkpoint activation. Moreover, co-fractionation experiments suggest that Mad2, Mad3 and Bub3 may be concomitantly present in protein complexes with Cdc20. Formation of the Bub3 Cdc20 complex requires all kinetochore checkpoint proteins but, surprisingly, not intact kinetochores. Conversely, point mutations altering the conserved WD40 motifs of Bub3, which might be involved in the formation of a beta-propeller fold devoted to protein-protein interactions, disrupt its association with Mad2, Mad3 and Cdc20, as well as proper checkpoint response. We suggest that Bub3 could serve as a platform for interactions between kinetochore checkpoint proteins, and its association with Mad2, Mad3 and Cdc20 might be instrumental for checkpoint activation. PMID- 11726502 TI - Fission yeast Rad50 stimulates sister chromatid recombination and links cohesion with repair. AB - To study the role of Rad50 in the DNA damage response, we cloned and deleted the Schizosaccharomyces pombe RAD50 homologue. The deletion is sensitive to a range of DNA-damaging agents and shows dynamic epistatic interactions with other recombination-repair genes. We show that Rad50 is necessary for recombinational repair of the DNA lesion at the mating-type locus and that rad50Delta shows slow DNA replication. We also find that Rad50 is not required for slowing down S phase in response to hydroxy urea or methyl methanesulfonate (MMS) treatment. Interestingly, in rad50Delta cells, the recombination frequency between two homologous chromosomes is increased at the expense of sister chromatid recombination. We propose that Rad50, an SMC-like protein, promotes the use of the sister chromatid as the template for homologous recombinational repair. In support of this, we found that Rad50 functions in the same pathway for the repair of MMS-induced damage as Rad21, the homologue of the Saccharomyces cerevisiae Scc1 cohesin protein. We speculate that Rad50 interacts with the cohesin complex during S phase to assist repair and possibly re-initiation of replication after replication fork collapse. PMID- 11726503 TI - p27 cytoplasmic localization is regulated by phosphorylation on Ser10 and is not a prerequisite for its proteolysis. AB - The activity of the cyclin-dependent kinase inhibitor p27 is controlled by its concentration and subcellular localization. However, the mechanisms that regulate its intracellular transport are poorly understood. Here we show that p27 is phosphorylated on Ser10 in vivo and that mutation of Ser10 to Ala inhibits p27 cytoplasmic relocalization in response to mitogenic stimulation. In contrast, a fraction of wild-type p27 and a p27(S10D)-phospho-mimetic mutant translocates to the cytoplasm in the presence of mitogens. G1 nuclear export of p27 and its Ser10 phosphorylation precede cyclin-dependent kinase 2 (Cdk2) activation and degradation of the bulk of p27. Interestingly, leptomycin B-mediated nuclear accumulation accelerates the turnover of endogenous p27; the p27(S10A) mutant, which is trapped in the nucleus, has a shorter half-life than wild-type p27 and the p27(S10D) mutant. In summary, p27 is efficiently degraded in the nucleus and phosphorylation of Ser10 is necessary for the nuclear to cytoplasmic redistribution of a fraction of p27 in response to mitogenic stimulation. This cytoplasmic localization may serve to decrease the abundance of p27 in the nucleus below a certain threshold required for activation of cyclin-Cdk2 complexes. PMID- 11726504 TI - Yeast prion protein derivative defective in aggregate shearing and production of new 'seeds'. AB - According to the nucleated polymerization model, in vivo prion proliferation occurs via dissociation (shearing) of the huge prion polymers into smaller oligomeric 'seeds', initiating new rounds of prion replication. Here, we identify the deletion derivative of yeast prion protein Sup35 (Sup35-Delta22/69) that is specifically defective in aggregate shearing and 'seed' production. This derivative, [PSI+], previously thought to be unable to turn into a prion state, in fact retains the ability to form a prion ([PSI+](Delta22/69)) that can be maintained in selective conditions and transmitted by cytoplasmic infection (cytoduction), but which is mitotically unstable in non-selective conditions. MorePSI+](Delta22/69) retains its mitotic stability defect. The [PSI+](Delta22/69) cells contain more Sup35 protein in the insoluble fraction and form larger Sup35 aggregates compared with the conventional [PSI+] cells. Moderate excess of Hsp104 disaggregase increases transmission of the [PSI+](Delta22/69) prion, while excess Hsp70-Ssa chaperone antagonizes it, opposite to their effects on conventional [PSI+]. Our results shed light on the mechanisms determining the differences between transmissible prions and non transmissible protein aggregates. PMID- 11726505 TI - Glycosylation influences cross-species formation of protease-resistant prion protein. AB - A key event in the transmissible spongiform encephalopathies (TSEs) is the formation of aggregated and protease-resistant prion protein, PrP-res, from a normally soluble, protease-sensitive and glycosylated precursor, PrP-sen. While amino acid sequence similarity between PrP-sen and PrP-res influences both PrP res formation and cross-species transmission of infectivity, the influence of co- or post-translational modifications to PrP-sen is unknown. Here we report that, if PrP-sen and PrP-res are derived from different species, PrP-sen glycosylation can significantly affect PrP-res formation. Glycosylation affected PrP-res formation by influencing the amount of PrP-sen bound to PrP-res, while the amino acid sequence of PrP-sen influenced the amount of PrP-res generated in the post binding conversion step. Our results show that in addition to amino acid sequence, co- or post-translational modifications to PrP-sen influence PrP-res formation in vitro. In vivo, these modifications might contribute to the resistance to infection associated with transmission of TSE infectivity across species barriers. PMID- 11726506 TI - A novel CCCH protein which modulates differentiation of Trypanosoma brucei to its procyclic form. AB - Cell differentiation in Trypanosoma brucei involves highly regulated changes in morphology, proliferation and metabolism. However, the controls of these developmental processes are unknown. We have identified two novel proteins from the rare CCCH zinc finger family, each <140 amino acids in length and implicated in life cycle regulation. TbZFP1 is transiently enriched during differentiation from the bloodstream to procyclic form, whereas tbZFP2, when ablated in bloodstream forms by RNA interference, inhibits this developmental step. Moreover, expressing an ectopic copy of tbZFP2 results in a dramatic procyclic stage-specific remodelling of the trypanosome cytoskeleton similar to the morphogenic events of differentiation. This phenotype, we term 'nozzle', involves polar extension of microtubules at the posterior end of the cell and is dependent upon a motif hitherto restricted to E3 ubiquitin ligases. TbZFP1 and tbZFP2 represent the first molecules implicated in the control of trypanosome differentiation to the procyclic form. PMID- 11726507 TI - Nuclear export of phosphorylated C/EBPbeta mediates the inhibition of albumin expression by TNF-alpha. AB - Decreased albumin expression is a frequent feature of cachexia patients afflicted with chronic diseases, including cancer, and a major contributor to their morbidity. Here we show that tumor necrosis-alpha (TNF-alpha) treatment of primary mouse hepatocytes or TNF-alpha overexpression in a mouse model of cachexia induces oxidative stress, nitric oxide synthase (NOS) expression and phosphorylation of C/EBPbeta on Ser239, within the nuclear localization signal, thus inducing its nuclear export, which inhibits transcription from the albumin gene. SIN-1, a NO donor, duplicated the TNF-alpha effects on hepatocytes. We found similar molecular abnormalities in the liver of patients with cancer cachexia. The cytoplasmic localization and association of C/EBPbeta-PSer239 with CRM1 (exportin-1) in TNF-alpha-treated hepatocytes was inhibited by leptomycin B, a blocker of CRM1 activity. Hepatic cells expressing the non-phosphorylatable C/EBPbeta alanine mutant were refractory to the inhibitory effects of TNF-alpha on albumin transcription since the mutant remained localized to the nucleus. Treatment of TNF-alpha mice with antioxidants or NOS inhibitors prevented phosphorylation of C/EBPbeta on Ser239 and its nuclear export, and rescued the abnormal albumin gene expression. PMID- 11726508 TI - Evidence for a novel GTPase priming step in the SRP protein targeting pathway. AB - Protein targeting by the signal recognition particle (SRP) pathway requires the interaction of two homologous GTPases that reciprocally regulate each other's GTPase activity, the SRP signal peptide- binding subunit (SRP54) and the SRP receptor alpha-subunit (SRalpha). The GTPase domain of both proteins abuts a unique 'N domain' that appears to facilitate external ligand binding. To examine the relationship between the unusual regulation and unique architecture of the SRP pathway GTPases, we mutated an invariant glycine in Escherichia coli SRP54 and SRalpha orthologs ('Ffh' and 'FtsY', respectively) that resides at the N GTPase domain interface. A G257A mutation in Ffh produced a lethal phenotype. The mutation did not significantly affect Ffh function, but severely reduced interaction with FtsY. Likewise, mutation of FtsY Gly455 produced growth defects and inhibited interaction with Ffh. The data suggest that Ffh and FtsY interact only in a 'primed' conformation which requires interdomain communication. Based on these results, we propose that the distinctive features of the SRP pathway GTPases evolved to ensure that SRP and the SR engage external ligands before interacting with each other. PMID- 11726509 TI - Involvement of the twin-arginine translocation system in protein secretion via the type II pathway. AB - The general secretory pathway (GSP) is a two-step process for the secretion of proteins by Gram-negative bacteria. The translocation across the outer membrane is carried out by the type II system, which involves machinery called the secreton. This step is considered to be an extension of the general export pathway, i.e. the export of proteins across the inner membrane by the Sec machinery. Here, we demonstrate that two substrates for the Pseudomonas aeruginosa secreton, both phospholipases, use the twin-arginine translocation (Tat) system, instead of the Sec system, for the first step of translocation across the inner membrane. These results challenge the previous vision of the GSP and suggest for the first time a mosaic model in which both the Sec and the Tat systems feed substrates into the secreton. Moreover, since P.aeruginosa phospholipases are secreted virulence factors, the Tat system appears to be a novel determinant of bacterial virulence. PMID- 11726510 TI - An acidic sequence of a putative yeast Golgi membrane protein binds COPII and facilitates ER export. AB - We previously identified Sys1p as a high copy number suppressor of Ypt6 GTPase deficient yeast mutants that are defective in endosome-to-Golgi transport. Here, we show that Sys1p is an integral membrane protein that resides on a post endoplasmic reticulum (ER) organelle(s). Affinity studies with detergent- solubilized yeast proteins showed that the C-terminal 53 amino acid tail of Sys1p binds effectively to the cytoplasmic Sec23p-Sec24p COPII subcomplex. This binding required a di-acidic Asp-Leu-Glu (DXE) motif, previously shown to mediate efficient ER export of the vesicular stomatitis virus glycoprotein in mammalian cells. In Sys1p, a Glu-Leu-Glu (EXE) sequence could not substitute for the (DXE) motif. Mutations of the (DXE) sequence resulted in ER retention of approximately 30% of the protein at steady state, whereas addition of the Sys1p tail to an ER resident membrane protein led to an intracellular redistribution of the chimeric protein. Our study demonstrates for the first time that, in yeast, a di-acidic sequence motif can act as a sorting signal for cargo selection during the formation of transport vesicles at the ER by direct binding to COPII component(s). PMID- 11726511 TI - Recruitment to Golgi membranes of ADP-ribosylation factor 1 is mediated by the cytoplasmic domain of p23. AB - Binding to Golgi membranes of ADP ribosylation factor 1 (ARF1) is the first event in the initiation of COPI coat assembly. Based on binding studies, a proteinaceous receptor has been proposed to be critical for this process. We now report that p23, a member of the p24 family of Golgi-resident transmembrane proteins, is involved in ARF1 binding to membranes. Using a cross-link approach based on a photolabile peptide corresponding to the cytoplasmic domain of p23, the GDP form of ARF1 (ARF1-GDP) is shown to interact with p23 whereas ARF1-GTP has no detectable affinity to p23. The p23 binding is shown to localize specifically to a 22 amino acid C-terminal fragment of ARF1. While a monomeric form of a non-photolabile p23 peptide does not significantly inhibit formation of the cross-link product, the corresponding dimeric form does compete efficiently for this interaction. Consistently, the dimeric p23 peptide strongly inhibits ARF1 binding to native Golgi membranes suggesting that an oligomeric form of p23 acts as a receptor for ARF1 before nucleotide exchange takes place. PMID- 11726512 TI - Two distinct effects on neurotransmission in a temperature-sensitive SNAP-25 mutant. AB - Vesicle fusion in eukaryotic cells is mediated by SNAREs (soluble N ethylmaleimide-sensitive factor attachment protein receptors). In neurons, the t SNARE SNAP-25 is essential for synaptic vesicle fusion but its exact role in this process is unknown. We have isolated a SNAP-25 temperature-sensitive paralytic mutant in Drosophila, SNAP-25(ts). The mutation causes a Gly50 to Glu change in SNAP-25's first amphipathic helix. A similar mutation in the yeast homologue SEC9 also results in temperature sensitivity, implying a conserved role for this domain in secretion. In vitro-generated 70 kDa SNARE complexes containing SNAP 25(ts) are thermally stable but the mutant SNARE multimers (of approximately 120 kDa) rapidly dissociate at 37 degrees C. The SNAP-25(ts) mutant has two effects on neurotransmitter release depending upon temperature. At 22 degrees C, evoked release of neurotransmitter in SNAP-25(ts) larvae is greatly increased, and at 37 degrees C, the release of neurotransmitter is reduced as compared with controls. Our data suggest that at 22 degrees C the mutation causes the SNARE complex to be more fusion competent but, at 37 degrees C the same mutation leads to SNARE multimer instability and fusion incompetence. PMID- 11726513 TI - Calreticulin and calnexin in the endoplasmic reticulum are important for phagocytosis. AB - Calreticulin and calnexin are Ca2+-binding proteins with chaperone activity in the endoplasmic reticulum. These proteins have been eliminated by gene replacement in Dictyostelium, the only microorganism known to harbor both proteins; family members in Dictyostelium are located at the base of phylogenetic trees. A dramatic decline in the rate of phagocytosis was observed in double mutants lacking calreticulin and calnexin, whereas only mild changes occurred in single mutants. Dictyostelium cells are professional phagocytes, capable of internalizing particles by a sequence of activities: adhesion of the particle to the cell surface, actin-dependent outgrowth of a phagocytic cup, and separation of the phagosome from the plasma membrane. In the double-null mutants, particles still adhered to the cell surface, but the outgrowth of phagocytic cups was compromised. Green fluorescent protein-tagged calreticulin and calnexin, expressed in wild-type cells, revealed a direct link of the endoplasmic reticulum to the phagocytic cup enclosing a particle, such that the Ca2+ storage capacity of calreticulin and calnexin might directly modulate activities of the actin system during particle uptake. PMID- 11726514 TI - Sphingoid base signaling via Pkh kinases is required for endocytosis in yeast. AB - In yeast, sphingoid base synthesis is required for the internalization step of endocytosis and organization of the actin cytoskeleton. We show that overexpression of either one of the two kinases Pkh1p or Pkh2p, that are homologous to mammalian 3-phosphoinositide-dependent kinase-1 (PDK1), can specifically suppress the sphingoid base synthesis requirement for endocytosis. Pkh1p and Pkh2p have an overlapping function because only a mutant with impaired function of both kinases is defective for endocytosis. Pkh1/2p kinases are activated in vitro by nanomolar concentrations of sphingoid base. These results suggest that Pkh1/2p kinases are part of a sphingoid base-mediated signaling pathway that is required for the internalization step of endocytosis. The Pkc1p kinase that is phosphorylated by Pkh1/2p kinases and plays a role in endocytosis was identified as one of the downstream effectors of this signaling cascade. PMID- 11726515 TI - Tyrosine phosphorylation of Grb2 by Bcr/Abl and epidermal growth factor receptor: a novel regulatory mechanism for tyrosine kinase signaling. AB - Growth factor receptor-binding protein-2 (Grb2) plays a key role in signal transduction initiated by Bcr/Abl oncoproteins and growth factors, functioning as an adaptor protein through its Src homology 2 and 3 (SH2 and SH3) domains. We found that Grb2 was tyrosine-phosphorylated in cells expressing BCR/ABL and in A431 cells stimulated with epidermal growth factor (EGF). Phosphorylation of Grb2 by Bcr/Abl or EGF receptor reduced its SH3-dependent binding to Sos in vivo, but not its SH2-dependent binding to Bcr/Abl. Tyr209 within the C-terminal SH3 domain of Grb2 was identified as one of the tyrosine phosphorylation sites, and phosphorylation of Tyr209 abolished the binding of the SH3 domain to a proline rich Sos peptide in vitro. In vivo expression of a Grb2 mutant where Tyr209 was changed to phenylalanine enhanced BCR/ABL-induced ERK activation and fibroblast transformation, and potentiated and prolonged Grb2-mediated activation of Ras, mitogen-activated protein kinase and c-Jun N-terminal kinase in response to EGF stimulation. These results suggest that tyrosine phosphorylation of Grb2 is a novel mechanism of down-regulation of tyrosine kinase signaling. PMID- 11726516 TI - Retroviral oncoprotein Tax induces processing of NF-kappaB2/p100 in T cells: evidence for the involvement of IKKalpha. AB - IkappaB kinase (IKK) is a key mediator of NF-kappaB activation induced by various immunological signals. In T cells and most other cell types, the primary target of IKK is a labile inhibitor of NF-kappaB, IkappaBalpha, which is responsible for the canonical NF-kappaB activation. Here, we show that in T cells infected with the human T-cell leukemia virus (HTLV), IKKalpha is targeted to a novel signaling pathway that mediates processing of the nfkappab2 precursor protein p100, resulting in active production of the NF-kappaB subunit, p52. This pathogenic action is mediated by the HTLV-encoded oncoprotein Tax, which appears to act by physically recruiting IKKalpha to p100, triggering phosphorylation-dependent ubiquitylation and processing of p100. These findings suggest a novel mechanism by which Tax modulates the NF-kappaB signaling pathway. PMID- 11726517 TI - CBP/p300 and muscle differentiation: no HAT, no muscle. AB - Terminal differentiation of muscle cells follows a precisely orchestrated program of transcriptional regulatory events at the promoters of both muscle-specific and ubiquitous genes. Two distinct families of transcriptional co-activators, GCN5/PCAF and CREB-binding protein (CBP)/p300, are crucial to this process. While both possess histone acetyl-transferase (HAT) activity, previous studies have failed to identify a requirement for CBP/p300 HAT function in myogenic differentiation. We have addressed this issue directly using a chemical inhibitor of CBP/p300 in addition to a negative transdominant mutant. Our results clearly demonstrate that CBP/p300 HAT activity is critical for myogenic terminal differentiation. Furthermore, this requirement is restricted to a subset of events in the differentiation program: cell fusion and specific gene expression. These data help to define the requirements for enzymatic function of distinct coactivators at different stages of the muscle cell differentiation program. PMID- 11726518 TI - T7 promoter release mediated by DNA scrunching. AB - Transcription initiation includes a phase in which short transcripts dissociate from the transcription complex and the polymerase appears not to move away from the promoter. During this process DNA may scrunch within the complex or the polymerase may transiently break promoter contacts to transcribe downstream DNA. Promoter release allowing extended downstream movement of the polymerase may be caused by RNA-mediated disruption of promoter contacts, or by limits on the amount of DNA that can be scrunched. Using exonuclease and KMnO4 footprinting of T7RNAP transcription complexes we show that the DNA scrunches during progression through initial transcription. To determine whether promoter release is determined by RNA length or by the amount of DNA scrunched, we compared release at promoters where the polymerase is forced to initiate at +2 with those where it initiates at +1. For RNAs of identical length, release is greater when more DNA is scrunched. Release is inhibited when a nick introduced into the template relieves the strain of scrunching. DNA scrunching therefore makes an important contribution to T7 promoter release. PMID- 11726519 TI - Functional interaction of STAT5 and nuclear receptor co-repressor SMRT: implications in negative regulation of STAT5-dependent transcription. AB - Signal transducers and activators of transcription (STATs) play a central role in cytokine signaling. Activating and repressing gene transcription is a dynamic process involving chromatin remodeling by histone acetylases and deacetylases, yet the role of this process in STAT-dependent transcription remains largely unknown. In a search for STAT5-interacting proteins by yeast two-hybrid screening, we identified the nuclear receptor co-repressor SMRT (silencing mediator for retinoic acid receptor and thyroid hormone receptor) as a potential STAT5-binding partner. SMRT binds to both STAT5A and 5B, and strongly repressed STAT5-dependent transcription in vitro. SMRT binds to the N-terminal coiled-coil domain of STAT5 and a mutation within this region previously found to render STAT5 hyperactive in response to cytokines abolished the interaction with SMRT. Overexpression of SMRT suppressed the induction of STAT5 target genes by interleukin-3, whereas the histone deacetylase inhibitor trichostatin A effectively enhanced and prolonged their expression. Together, these findings illuminate the potential role of SMRT in down-regulating STAT5 activity, with a consequent reduction of STAT5 target gene expression. PMID- 11726520 TI - PNPase autocontrols its expression by degrading a double-stranded structure in the pnp mRNA leader. AB - Polynucleotide phosphorylase synthesis is autocontrolled at a post transcriptional level in an RNase III-dependent mechanism. RNase III cleaves a long stem-loop in the pnp leader, which triggers pnp mRNA instability, resulting in a decrease in the synthesis of polynucleotide phosphorylase. The staggered cleavage by RNase III removes the upper part of the stem-loop structure, creating a duplex with a short 3' extension. Mutations or high temperatures, which destabilize the cleaved stem-loop, decrease expression of pnp, while mutations that stabilize the stem increase expression. We propose that the dangling 3' end of the duplex created by RNase III constitutes a target for polynucleotide phosphorylase, which binds to and degrades the upstream half of this duplex, hence inducing pnp mRNA instability. Consistent with this interpretation, a pnp mRNA starting at the downstream RNase III processing site exhibits a very low level of expression, regardless of the presence of polynucleotide phosphorylase. Moreover, using an in vitro synthesized pnp leader transcript, it is shown that polynucleotide phosphorylase is able to digest the duplex formed after RNase III cleavage. PMID- 11726521 TI - Release of U18 snoRNA from its host intron requires interaction of Nop1p with the Rnt1p endonuclease. AB - An external stem, essential for the release of small nucleolar RNAs (snoRNAs) from their pre-mRNAs, flanks the majority of yeast intron-encoded snoRNAs. Even if this stem is not a canonical Rnt1p substrate, several experiments have indicated that the Rnt1p endonuclease is required for snoRNA processing. To identify the factors necessary for processing of intron-encoded snoRNAs, we have raised in vitro extracts able to reproduce such activity. We found that snoRNP factors are associated with the snoRNA- coding region throughout all the processing steps, and that mutants unable to assemble snoRNPs have a processing deficient phenotype. Specific depletion of Nop1p completely prevents U18 snoRNA synthesis, but does not affect processing of a dicistronic snoRNA-coding unit that has a canonical Rnt1p site. Correct cleavage of intron-encoded U18 and snR38 snoRNAs can be reproduced in vitro by incubating together purified Nop1p and Rnt1p. Pull-down experiments showed that the two proteins interact physically. These data indicate that cleavage of U18, snR38 and possibly other intron-encoded snoRNAs is a regulated process, since the stem is cleaved by the Rnt1p endonuclease only when snoRNP assembly has occurred. PMID- 11726522 TI - Control of branch-site choice by a group II intron. AB - The branch site of group II introns is typically a bulged adenosine near the 3' end of intron domain 6. The branch site is chosen with extraordinarily high fidelity, even when the adenosine is mutated to other bases or if the typically bulged adenosine is paired. Given these facts, it has been difficult to discern the mechanism by which the proper branch site is chosen. In order to dissect the determinants for branch-point recognition, new mutations were introduced in the vicinity of the branch site and surrounding domains. Single mutations did not alter the high fidelity for proper branch-site selection. However, several combinations of mutations moved the branch site systematically to new positions along the domain 6 stem. Analysis of those mutants, together with a new alignment of domain 5 and domain 6 sequences, reveals a set of structural determinants that appear to govern branch-site selection by group II introns. PMID- 11726523 TI - Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. AB - Duplexes of 21-23 nucleotide (nt) RNAs are the sequence-specific mediators of RNA interference (RNAi) and post-transcriptional gene silencing (PTGS). Synthetic, short interfering RNAs (siRNAs) were examined in Drosophila melanogaster embryo lysate for their requirements regarding length, structure, chemical composition and sequence in order to mediate efficient RNAi. Duplexes of 21 nt siRNAs with 2 nt 3' overhangs were the most efficient triggers of sequence-specific mRNA degradation. Substitution of one or both siRNA strands by 2'-deoxy or 2'-O-methyl oligonucleotides abolished RNAi, although multiple 2'-deoxynucleotide substitutions at the 3' end of siRNAs were tolerated. The target recognition process is highly sequence specific, but not all positions of a siRNA contribute equally to target recognition; mismatches in the centre of the siRNA duplex prevent target RNA cleavage. The position of the cleavage site in the target RNA is defined by the 5' end of the guide siRNA rather than its 3' end. These results provide a rational basis for the design of siRNAs in future gene targeting experiments. PMID- 11726524 TI - The intracellular location of two aminoacyl-tRNA synthetases depends on complex formation with Arc1p. AB - In yeast, two aminoacyl-tRNA synthetases, MetRS and GluRS, are associated with Arc1p. We have studied the mechanism of this complex formation and found that the non-catalytic N-terminally appended domains of MetRS and GluRS are necessary and sufficient for binding to Arc1p. Similarly, it is the N-terminal domain of Arc1p that contains distinct but overlapping binding sites for MetRS and GluRS. Localization of Arc1p, MetRS and GluRS in living cells using green fluorescent protein showed that these three proteins are cytoplasmic and largely excluded from the nucleus. However, when their assembly into a complex is inhibited, significant amounts of MetRS, GluRS and Arc1p can enter the nucleus. We suggest that the organization of aminoacyl-tRNA synthetases into a multimeric complex not only affects catalysis, but is also a means of segregating the tRNA- aminoacylation machinery mainly to the cytoplasmic compartment. PMID- 11726525 TI - Cell-specific proteins regulate viral RNA translation and virus-induced disease. AB - Translation initiation of the picornavirus genome is regulated by an internal ribosome entry site (IRES). The IRES of a neurovirulent picornavirus, the GDVII strain of Theiler's murine encephalomyelitis virus, requires polypyrimidine tract binding protein (PTB) for its function. Although neural cells are deficient in PTB, they express a neural-specific homologue of PTB (nPTB). We now show that nPTB and PTB bind similarly to multiple sites in the GDVII IRES, rendering it competent for efficient translation initiation. Mutation of a PTB or nPTB site results in a more prominent decrease in nPTB than PTB binding, a decrease in activity of nPTB compared with PTB in promoting translation initiation, and attenuation of the neurovirulence of the virus without a marked effect on virus growth in non-neural cells. The addition of a second-site mutation in the mutant IRES generates a new PTB (nPTB) binding site, and restores nPTB binding, translation initiation and neurovirulence. We conclude that the tissue-specific expression and differential RNA-binding properties of PTB and nPTB are important determinants of cell-specific translational control and viral neurovirulence. PMID- 11726526 TI - Heme-regulated eIF2alpha kinase (HRI) is required for translational regulation and survival of erythroid precursors in iron deficiency. AB - Although the physiological role of tissue-specific translational control of gene expression in mammals has long been suspected on the basis of biochemical studies, direct evidence has been lacking. Here, we report on the targeted disruption of the gene encoding the heme-regulated eIF2alpha kinase (HRI) in mice. We establish that HRI, which is expressed predominantly in erythroid cells, regulates the synthesis of both alpha- and beta-globins in red blood cell (RBC) precursors by inhibiting the general translation initiation factor eIF2. This inhibition occurs when the intracellular concentration of heme declines, thereby preventing the synthesis of globin peptides in excess of heme. In iron-deficient HRI(-/-) mice, globins devoid of heme aggregated within the RBC and its precursors, resulting in a hyperchromic, normocytic anemia with decreased RBC counts, compensatory erythroid hyperplasia and accelerated apoptosis in bone marrow and spleen. Thus, HRI is a physiological regulator of gene expression and cell survival in the erythroid lineage. PMID- 11726528 TI - Conformational isomerization in phage Mu transpososome assembly: effects of the transpositional enhancer and of MuB. AB - Initiation of phage Mu DNA transposition requires assembly of higher order protein-DNA complexes called Mu transpososomes containing the two Mu DNA ends and MuA transposase tetramer. Mu transpososome assembly is highly regulated and involves multiple DNA sites for transposase binding, including a transpositional enhancer called the internal activation sequence (IAS). In addition, a number of protein cofactors participate, including the target DNA activator MuB ATPase. We investigated the impact of the assembly cofactors on the kinetics of transpososome assembly with the aim of deciphering the reaction steps that are influenced by the cofactors. The transpositional enhancer IAS appears to have little impact on the initial pairing of the two Mu end segments bound by MuA. Instead, it accelerates the post-synaptic conformational step(s) that converts the reversible complex to the stable transpososome. The transpososome assembly stimulation by MuB does not require its stable DNA binding activity, which appears critical for directing transposition to sites distant from the donor transposon. PMID- 11726529 TI - Nutrients as regulators of immune function: Introduction. PMID- 11726527 TI - DNA polymerase beta is the major dRP lyase involved in repair of oxidative base lesions in DNA by mammalian cell extracts. AB - The repair of oxidative base lesions in DNA is a coordinated chain of reactions that includes removal of the damaged base, incision of the phosphodiester backbone at the abasic sugar residue, incorporation of an undamaged nucleotide and sealing of the DNA strand break. Although removal of a damaged base in mammalian cells is initiated primarily by a damage-specific DNA glycosylase, several lyases and DNA polymerases may contribute to the later stages of repair. DNA polymerase beta (Pol beta) was implicated recently as the major polymerase involved in repair of oxidative base lesions; however, the identity of the lyase participating in the repair of oxidative lesions is unclear. We studied the mechanism by which mammalian cell extracts process DNA substrates containing a single 8-oxoguanine or 5,6-dihydrouracil at a defined position. We find that, when repair synthesis proceeds through a Pol beta-dependent single nucleotide replacement mechanism, the 5'-deoxyribosephosphate lyase activity of Pol beta is essential for repair of both lesions. PMID- 11726530 TI - Modulation of the expression of cyclooxygenase-2 by fatty acids mediated through toll-like receptor 4-derived signaling pathways. AB - Genetic evidence that Toll-like receptor 4 (Tlr4) is the lipopolysaccharide (LPS) receptor and biochemical evidence that Tlr4 confers LPS responsiveness as determined by activation of NF-kappaB and expression of inducible cyclooxygenase 2 have been demonstrated. Saturated fatty acids (SFAs) acylated in lipid A moiety of LPS are essential for biological activities of LPS. It is now demonstrated that SFAs, but not unsaturated fatty acids (UFAs), induce NF-kappaB activation and expression of COX-2 and other inflammatory markers in macrophages. UFAs inhibit COX-2 expression induced by SFAs and LPS. Additional evidence suggests that both SFA-induced COX-2 expression and its inhibition by UFAs are mediated through a common signaling pathway derived from Tlr4. These results represent a novel mechanism by which fatty acids modulate signaling pathways and target gene expression. Whether fatty acids also modulate signaling pathways and target gene expression derived from the activation of other Tlrs remains to be determined. Hwang, D. Modulation of the expression of cyclooxygenase 2 by fatty acids mediated through Toll-like receptor 4-derived signaling pathways. PMID- 11726531 TI - Leptin regulation of the immune response and the immunodeficiency of malnutrition. AB - Leptin is a 16 kDa protein mainly produced by adipose tissue in proportion to adipose tissue mass. Originally thought to be a satiety factor, leptin is a pleiotropic molecule. In addition to playing a role in energy regulation, leptin also regulates endocrine and immune functions. Both the structure of leptin and that of its receptor suggest that leptin might be classified as a cytokine. The secondary structure of leptin has similarities to the long-chain helical cytokines family, which includes interleukin 6 (IL-6), IL-11, CNTF, and LIF, and the leptin receptor is homologous to the gp-130 signal-transducing subunit of the IL-6-type cytokine receptors. Leptin plays a role in innate and acquired immunity. Leptin levels increase acutely during infection and inflammation, and may represent a protective component of the host response to inflammation. More important, leptin deficiency increases susceptibility to infectious and inflammatory stimuli and is associated with dysregulation of cytokine production. Leptin deficiency also causes a defect in hematopoiesis. Leptin regulates T cells responses, polarizing Th cells toward a Th1 phenotype. Low leptin levels occurring during starvation mediate the neuroendocrine and immune dysfunction of starvation. PMID- 11726532 TI - A distinct role for apoptosis in the changes in lymphopoiesis and myelopoiesis created by deficiencies in zinc. AB - Reduced numbers of lymphocytes in the peripheral immune system appeared to be a significant cause of the loss in host defense capacity in humans and animals that are zinc deficient (ZD). A series of studies verified that ZD substantially reduced the lymphocyte compartment of both the marrow and thymus in young adult mice, with large losses noted among the pre-B and pre-T cells. Suboptimal nutriture along with chronic production of glucocorticoids generated during ZD had accelerated apoptosis among these precursor lymphocytes two- to threefold. Thus, the primary cause of the lymphopenia created by ZD was reduced production of lymphocytes and heightened cell death among precursor cells. The data will also show that myelopoiesis in the marrow was protected and enhanced numbers of myeloid progenitor cells were found in S and G2/M. Thus, as zinc became limiting the second line of defense appeared to be down-regulated via reduction of lymphopoiesis while cells of the myeloid lineage were protected to maintain the first line of defense that provides innate immunity. This may represent an important adaptation of the immune system to suboptimal nutriture that deserves further exploration. PMID- 11726533 TI - Vitamin D: its role and uses in immunology. AB - In recent years there has been an effort to understand possible noncalcemic roles of vitamin D, including its role in the immune system and, in particular, on T cell-medicated immunity. Vitamin D receptor is found in significant concentrations in the T lymphocyte and macrophage populations. However, its highest concentration is in the immature immune cells of the thymus and the mature CD-8 T lymphocytes. The significant role of vitamin D compounds as selective immunosuppressants is illustrated by their ability to either prevent or markedly suppress animal models of autoimmune disease. Results show that 1,25 dihydroxyvitamin D3 can either prevent or markedly suppress experimental autoimmune encephalomyelitis, rheumatoid arthritis, systemic lupus erythematosus, type I diabetes, and inflammatory bowel disease. In almost every case, the action of the vitamin D hormone requires that the animals be maintained on a normal or high calcium diet. Possible mechanisms of suppression of these autoimmune disorders by the vitamin D hormone have been presented. The vitamin D hormone stimulates transforming growth factor TGFbeta-1 and interleukin 4 (IL-4) production, which in turn may suppress inflammatory T cell activity. In support of this, the vitamin D hormone is unable to suppress a murine model of the human disease multiple sclerosis in IL-4-deficient mice. The results suggest an important role for vitamin D in autoimmune disorders and provide a fertile and interesting area of research that may yield important new therapies. PMID- 11726534 TI - Distinct myoprotective roles of cardiac sarcolemmal and mitochondrial KATP channels during metabolic inhibition and recovery. AB - The protective roles of sarcolemmal (sarc) and mitochondrial (mito) KATP channels are unclear despite their apparent importance to ischemic preconditioning. We examined these roles by monitoring intracellular calcium ([Ca]int), using fura-2 and fluo-3, in enzymatically isolated rat right ventricular myocytes. Myocyte mortality, estimated using a trypan blue assay, changed approximately in parallel with changes in [Ca]int. Chemically induced hypoxia (CIH), induced by application of cyanide and 2-deoxy-glucose, caused a steady rise in [Ca]int. Calcium increased more rapidly on 'reoxygenation' by return to control solutions. The protein kinase C (PKC) activator PMA abolished both phases of calcium increase. The mitoKATP channel-selective blocker 5-hydroxydecanoate partially prevented the PMA-induced protection during CIH, but not during reoxygenation. In contrast, HMR 1098, a sarcKATP channel-selective blocker, abolished protection only during the reoxygenation. Adenosine (A1) receptor activation prevented or reduced increases in [Ca]int and improved cell viability via a PKC and mito/sarcKATP channel dependent mechanism. PKC-dependent protection against cytoplasmic calcium increases was also observed in a human cell line (tsA201) transiently expressing sarcKATP channels. Protection was abolished only during the reoxygenation phase by the amino acid substitution (T180A) in the pore-forming Kir6.2 subunit, a mutation previously shown to prevent PKC-dependent modulation. Our data suggest that sarc and mitoKATP channel populations play distinct protective roles, triggered by PKC and/or adenosine, during chemically induced hypoxia/reoxygenation. PMID- 11726535 TI - Implication of acyl chain of diacylglycerols in activation of different isoforms of protein kinase C. AB - We synthesized diacylglycerols (DAGs) containing omega-6 or omega-3 polyunsaturated fatty acids [i.e., 1-stearoyl-2-arachidonoyl-sn-glycerol (SAG), 1 stearoyl-2-docosahexaenoyl-sn-glycerol (SDG), and 1-stearoyl-2-eicosapentaenoyl sn-glycerol (SEG)] and assessed their efficiency on activation of conventional (alpha, beta I, gamma) and novel (epsilon, delta) protein kinase C (PKC). SAG exerted significantly higher stimulatory effects than SDG and SEG on activation of PKC alpha and PKC delta. Activation of PKC beta I by SEG and SDG was higher than that by SAG. Activation of PKC gamma did not differ significantly among DAG molecular species. Addition of SAG to assays containing SEG and SDG exerted additive effects on activation of alpha and epsilon, but not on beta I and gamma, isoforms of PKC. SDG- and SEG-induced activation of PKC delta was significantly curtailed by the addition of SAG. Three DAG species significantly curtailed the PMA-induced activation of beta Iota, gamma, and delta, but not of alpha and epsilon, isoforms of PKC. Our study demonstrates for the first time that in vitro activation of different PKC isoenzymes vary in response to different DAG species, and one can envisage that this differential regulation may be responsible for their in vivo effects on target organs. PMID- 11726536 TI - Antibodies against the carboxyl-terminal end of the Trypanosoma cruzi ribosomal P proteins are pathogenic. AB - Sera from patients with chronic Chagas heart disease recognize the carboxyl terminal regions of the Trypanosoma cruzi ribosomal P proteins defined by B cell epitopes P013 (EDDDDDFGMGALF) and R13 (EEEDDDMGFGLFD) corresponding to the T. cruzi ribosomal P0 (TcP0) and P2beta (TcP2beta) proteins, respectively. It has been hypothesized that both epitopes may induce antibodies that cross-react and stimulate the beta1-adrenoreceptor. However, no proof as to their pathogenicity has been obtained. We investigated the consequences of immunizing mice with either TcP0 or TcP2beta proteins. Of 24 immunized animals, 16 generated antibodies against the carboxyl-terminal end of the corresponding protein, 13 of which showed an altered ECG (P<0.001, 81%). Immunization with TcP0 induced anti P013 antibodies that bind to and stimulate cardiac G-protein-coupled receptors and are linked to the induction of supraventricular arrhythmia, repolarization, and conduction abnormalities as monitored by serial electrocardiographic analysis. In contrast, immunization with TcP2beta generated anti-R13 antibodies with an exclusive beta1-adrenergic-stimulating activity whose appearance strictly correlated with the recording of supraventricular tachycardia and death. These findings demonstrate that anti-P antibodies are arrhythmogenic in the setting of a normal heart, since no inflammatory lesions or fibrosis were evident to light microscopic examination. PMID- 11726537 TI - Hypoxia affects expression of circadian genes PER1 and CLOCK in mouse brain. AB - The key elements of circadian clockwork and oxygen homeostasis are the PAS protein family members PER and CLOCK and hypoxia-inducible factor 1alpha (HIF 1alpha). The PAS domain serves as an interface for protein-protein interactions. We asked whether a cross-talk exists between the PAS components of hypoxic and circadian pathways. We found several isoforms of PER1 protein that exhibit tissue specific size differences. In the mouse brain, a predominantly nuclear 48 kDa isoform that followed a daily rhythm was observed. The 48 kDa form was found in the nuclear fractions derived from mouse liver, Swiss3T3 fibroblasts, and N2A neuroblastoma cells. In mouse kidney and human 293 kidney cells, a 55 kDa PER1 form was detected. CLOCK was observed as a predicted 100 kDa protein in rat-1 cells and in all analyzed mouse tissues including brain, liver, kidney, and spleen. In contrast to PER1, CLOCK protein expression was not rhythmic. Exposure to hypoxia led to increased PER1 and CLOCK protein levels in mice. Based on coimmunoprecipitation experiments that showed protein-protein interaction between PER1 and the alpha subunit of HIF-1, we suggest that these hypoxic effects may be modulated by HIF-1alpha.-Chilov, D., Hofer, T., Bauer, C., Wenger, R. H., Gassmann, M. Hypoxia affects expression of circadian genes PER1 and CLOCK in mouse brain. PMID- 11726538 TI - Pathophysiology of apolipoprotein E deficiency in mice: relevance to apo E related disorders in humans. AB - Apolipoprotein E (apo E) deficiency (or its abnormalities in humans) is associated with a series of pathological conditions including dyslipidemia, atherosclerosis, Alzheimer's disease, and shorter life span. The purpose of this study was to characterize these conditions in apo E-deficient C57BL/6J mice and relate them to human disorders. Deletion of apo E gene in mice is associated with changes in lipoprotein metabolism [plasma total cholesterol (TC) (>+400%), HDL cholesterol (-80%), HDL/TC, and HDL/LDL ratios (-93% and -96%, respectively), esterification rate in apo B-depleted plasma (+100%), plasma triglyceride (+200%), hepatic HMG-CoA reductase activity (-50%), hepatic cholesterol content (+30%)], decreased plasma homocyst(e)ine and glucose levels, and severe atherosclerosis and cutaneous xanthomatosis. Hepatic and lipoprotein lipase activities, hepatic LDL receptor function, and organ antioxidant capacity remain unchanged. Several histological/immunohistological stainings failed to detect potential markers for neurodegenerative disease in the brain of 37-wk-old male apo E-KO mice. Apo E-KO mice may have normal growth and development, but advanced atherosclerosis and xanthomatosis may indirectly reduce their life span. Apo E plays a crucial role in regulation of lipid metabolism and atherogenesis without affecting lipase activities, endogenous antioxidant capacity, or appearance of neurodegenerative markers in 37-wk-old male mice. PMID- 11726539 TI - DNA methylation-dependent chromatin fiber compaction in vivo and in vitro: requirement for linker histone. AB - Dynamic alterations in chromatin structure mediated by postsynthetic histone modifications and DNA methylation constitute a major regulatory mechanism in DNA functioning. DNA methylation has been implicated in transcriptional silencing, in part by inducing chromatin condensation. To understand the methylation-dependent chromatin structure, we performed atomic force microscope (AFM) studies of fibers isolated from cultured cells containing normal or elevated levels of m5C. Chromatin fibers were reconstituted on control or methylated DNA templates in the presence or absence of linker histone. Visual inspection of AFM images, combined with quantitative analysis of fiber structural parameters, suggested that DNA methylation induced fiber compaction only in the presence of linker histones. This conclusion was further substantiated by biochemical results. PMID- 11726540 TI - Effect of graded hypoxia on the rat hepatic tissue oxygenation and energy metabolism monitored by near-infrared and 31P nuclear magnetic resonance spectroscopy. AB - Alteration in hepatic cellular adenosine triphosphate (ATP) levels has been shown to be a sensitive index for hypoxic damage. Hepatic ATP metabolism can be monitored by 31P nuclear magnetic resonance (NMR). Near-infrared spectroscopy (NIRS) can measure tissue oxyhemoglobin (HbO2), deoxyhemoglobin (Hb), and cytochrome oxidase (Cyt Ox), which reflect ATP production. In this study, hepatic oxygenation parameters have been correlated with ATP metabolism under graded hypoxia. Sprague-Dawley rats underwent laparotomy for liver exposure. NIRS probes and an NMR coil were placed on the liver and the animal was positioned in the NMR magnet. Graded hypoxia was achieved by a stepwise reduction of the fraction of inspired oxygen (FiO2) from 15 to 4%. Recovery between the hypoxic periods was achieved using 30% oxygen. Hepatic tissue oxygenation parameters were measured continuously by NIRS; 31P-NMR spectra obtained at 1 min intervals from energy metabolites and intracellular pH were calculated. All the hypoxic grades produced an immediate reduction in HbO2 with a simultaneous increase in Hb. Cyt Ox was reduced significantly only with FiO2 of 45 kb) and producing abnormal DMPK mRNA with at least 300 CUG repeats, displayed clinical, histological, molecular and electrophysiological abnormalities in skeletal muscle consistent with those observed in DM1 patients. Like DM1 patients, these transgenic mice show abnormal tau expression in the brain. These results provide further evidence for the RNA trans-dominant effect of the CUG expansion, not only in muscle, but also in brain. PMID- 11726561 TI - Cost-effectiveness modelling. PMID- 11726560 TI - Regulation of murine survival motor neuron (Smn) protein levels by modifying Smn exon 7 splicing. AB - Proximal spinal muscular atrophy (SMA) is caused by mutations in the survival motor neuron gene (SMN1). In humans, two nearly identical copies of SMN exist and differ only by a single non-polymorphic C-->T nucleotide transition in exon 7. SMN1 contains a 'C' nucleotide at the +6 position of exon 7 and produces primarily full-length SMN transcripts, whereas SMN2 contains a 'T' nucleotide and produces high levels of a transcript that lacks exon 7 and a low level of full length SMN transcripts. All SMA patients lack a functional SMN1 gene but retain at least one copy of SMN2, suggesting that the low level of full-length protein produced from SMN2 is sufficient for all cell types except motor neurons. The murine Smn gene is not duplicated or alternatively spliced. It resembles SMN1 in that the critical exon 7 +6 'C' nucleotide is conserved. We have generated Smn minigenes containing either wild-type Smn exon 7 or an altered exon 7 containing the C-->T nucleotide transition to mimic SMN2. When expressed in cultured cells or transgenic mice, the wild-type minigene produced only full-length transcripts whereas the modified minigene alternatively spliced exon 7. Furthermore, Smn exon 7 contains a critical AG-rich exonic splice enhancer sequence (ESE) analogous to the human ESE within SMN exon 7, and subtle mutations within the mESE caused a variation in Smn transcript levels. In summary, we show for the first time that the murine Smn locus can be induced to alternatively splice exon 7. These results demonstrate that SMN protein levels can be varied in the mouse by the introduction of specific mutations at the endogenous Smn locus and thereby lay the foundation for developing animals that closely 'resemble' SMA patients. PMID- 11726562 TI - Investigation of the infertile couple: a one-stop ultrasound-based approach. AB - The appropriateness of many investigations for subfertility will continue to be of debate for some time yet. Of most benefit to the concerned couple would be a process that is diagnostically accurate, expeditious and reliable. It should be performed with a minimum of invasion and provide both patient and clinician with useful prognostic information regarding possible future treatment. This article is intended to illustrate the advantages of an ultrasound-based process of subfertility investigation. Discussed is the role of ultrasound compared with more invasive investigative methods such as laparoscopy and hysteroscopy. In addition, the potential capacity of newer advanced ultrasound technologies is reviewed. PMID- 11726563 TI - Should ICSI be the treatment of choice for all cases of in-vitro conception? Considerations of fertilization and embryo development, cost effectiveness and safety. AB - There is now considerable discussion whether intracytoplasmic sperm injection (ICSI) should be used in all cases of IVF. A critical and balanced view of the current literature is presented. The difficult question is how to identify men with apparently normal semen who are likely to fail to achieve a pregnancy using IVF. In conclusion, from both the safety and scientific viewpoint, ICSI should only be used in cases where success at IVF is regarded as unlikely. PMID- 11726564 TI - Preventive attitude of physicians to avoid OHSS in IVF patients. AB - The ovarian hyperstimulation syndrome (OHSS) is a dramatic iatrogenic complication of fertility treatment. There is no consensus about its prevention strategies. We assessed whether physicians modify their preventive attitude in relation to clinical risk factors and to the oestradiol response chart. Three case scenarios with three levels of risk factors for OHSS were constructed. For each case scenario, four different charts of the oestradiol curve were described (peak serum oestradiol of 3590 or 6590 pg/ml obtained after a shorter or longer period). At random, we sent three out of the 12 artificially constructed case scenarios to 573 physicians who are members of the European Society of Human Reproduction and Embryology (ESHRE). They were asked whether they would cancel the cycle, take preventive measures or proceed to a regular IVF. A total of 389 cases from 130 different physicians was analysed. Globally, in 23% of the cases the physicians would proceed to a regular IVF. This decision varied significantly according to the risk level (between 38% in low risk to 8% in high risk cases; P < 0.01) and in relation to the oestradiol curve (P < 0.01). In 11% of the cases they would cancel the cycle and in 66% take some preventive measures. Among the selected preventive measures, coasting was by far the most popular choice (60%), followed by the use of i.v. albumin or hydroxyaethyl starch solution (36%) and cryo-preservation of all embryos (33%). In view of the sparse data as to whether coasting is really an effective method, and on how it should be carried out, we suggest that there is an urgent need to evaluate coasting as a preventive method in a large randomized trial in order to properly assess its efficiency and to provide precise guidelines for its use. PMID- 11726565 TI - Model of counter-current transfer from vagina to urethra in postmenopausal women. AB - BACKGROUND: Low-dose vaginal oestrogens are effective in treating post-menopausal urogenital atrophy without inducing endometrial proliferation. We aimed to assess whether this dichotomic effect could be the result of a preferential vagina-to urethra transfer via a counter-current transfer of oestrogens from vagina to the arterial blood supplying the urethra. Due to the impossibility of obtaining blood samples from urethral arteries, and since the nature of counter-current exchange of substances is similar to the transfer of heat, we investigated cold transfer throughout the anterior vaginal wall to the vesical trigone and urethra. METHODS: Plastic tubes filled with cold saline were inserted into the vagina of 30 menopausal women. Temperatures were recorded at the vesical trigone and at three different urethral sites. Comparisons were performed 2 and 4.5 min after starting of cooling, and 4.5 min after removal of tubes. RESULTS: The urethra cooled significantly more than the bladder. Urethral cooling was not homogeneous; distal sites of the urethra cooled significantly more than proximal site and bladder despite a larger distance to the vaginal cooling device. CONCLUSIONS: Distribution of cold from the vagina to the urethra is not the result of simple diffusion and mechanisms of preferential distribution may exist from the vagina to the middle and low part of the urethra. PMID- 11726566 TI - Characteristics of human ovulation in natural cycles correlated with age and achievement of pregnancy. AB - BACKGROUND: The aim of this study was to evaluate whether characteristics of human ovulation correlate with age and pregnancy potential. METHODS: Two groups of women with regular menstrual cycles were included (i.e. one fertile and one infertile group), which were divided into four age groups (< or =29, 30-34, 35 39, > or = 40 years). Monitoring included observations of follicular phase length, whether ovulations occurred from the left or right ovary, the pattern of ovulations in succeeding natural cycles and, in a subset of women, early follicular phase FSH concentrations. RESULTS: Ovulation moving from one ovary to the other in two consecutive cycles (i.e. contralateral ovulation) was inversely correlated with age, showing a ratio of contralateral ovulation per contra plus ipsilateral ovulations (C/C+I) of 62% in women <29 years, gradually decreasing to 42% in women >40 years. The ratio of right-sided ovulation per right plus left sided ovulations (R/R+L) was unrelated to age and remained almost constant at a level of approximately 55%. The follicular phase length was inversely correlated with age, being 16.2 +/- 2.9, 15.4 +/- 2.9, 14.8 +/- 2.8 and 13.7 +/- 1.3 days in women < 29, 30-34, 35-39 and >40 years of age respectively. The follicular phase length was similar when comparing ovulations occurring from the right and left ovary, but comparing two successive cycles, the length of the follicular phase of the second cycle, showing contralateral ovulation, was shorter than ipsilateral ovulation with two consecutive ovulations in the same ovary. The pregnancy rate of the four groups decreased with age, being 14, 12, 5 and 3% respectively. The C/C+I ratio correlates with pregnancy rate and follicular phase length, and inversely correlates with basal FSH, whereas R/R+L is unrelated to age and pregnancy rate. CONCLUSIONS: Human ovulation shows characteristics related to age. The interaction between the two ovaries seems to be most pronounced in the younger years, where ovulations jump from one ovary to the other more frequently than later on in life. The C/C+I ratio shows a clear correlation with age and pregnancy rate. PMID- 11726567 TI - Follicular fluid responds endothermically to aqueous dilution. AB - BACKGROUND: Studies suggest that ovarian follicles are cooler than their surrounding tissues. The mechanism of this remarkable phenomenon is unclear. We postulate that endothermic reactions accompany the growth-associated hydration of follicular fluid. METHODS: We performed two types of experiment, using human and animal follicular fluids. In the first, saline (50 microl) was injected into follicular fluid (500 microl) held in an equilibrated incubator, with monitoring of sample temperature. In the second, an adiabatic microcalorimeter recorded thermal shifts after injection of buffer (10 microl) into previously dialysed samples (1.4 ml). The relevance of changes observed was assessed by mathematical modelling. RESULTS: In the incubator study, 9/17 bovine and 6/12 human fluids showed a temperature fall (0.05-0.2 degrees C). Cooling was delayed by up to 2 min but sustained for 7-25 min. Remaining fluids showed no change. In the microcalorimeter, 4/9 human, 4/6 bovine, 5/5 porcine and 1/4 equine samples showed an endothermic response. Remaining samples showed either no response (bovine) or exothermy (human, equine). Pre-concentration of human follicular fluid amplified the endothermy or reversed the exothermy. Modelling indicated that the incubator-type response was of appropriate magnitude to explain follicular hypothermy. CONCLUSION: Follicular fluid responds endothermically to aqueous dilution and may contribute to follicular cooling during growth. PMID- 11726568 TI - Angiogenesis in the human corpus luteum: simulated early pregnancy by HCG treatment is associated with both angiogenesis and vessel stabilization. AB - BACKGROUND: This study examined changes in the luteal vasculature throughout the menstrual cycle and during simulated pregnancy with human chorionic gonadotrophin (HCG) in the human. METHODS: Endothelial cell and pericyte area were assessed by quantitative immunocytochemistry for CD34 and alpha-smooth muscle actin respectively, taking into consideration the dynamics of lutein cell hypertrophy and atrophy throughout the cycle and after HCG treatment. Endothelial cell proliferation was detected by Ki-67/CD34 dual staining and a proliferation index was obtained. The molecular regulation of angiogenesis was studied by examining changes in vascular endothelial growth factor (VEGF) immunostaining. RESULTS: The early luteal phase is associated with intense angiogenesis, as indicated by high endothelial cell proliferation, and by the mid-luteal phase a mature vasculature was apparent, as shown by maximal endothelial cell and pericyte areas. During the late luteal phase, decreased endothelial proliferation, endothelial cell and pericyte area indicated vascular regression. HCG treatment induced a second burst of total and endothelial cell proliferation and a concomitant increase in endothelial cell and pericyte areas. VEGF protein was expressed throughout the luteal phase and a significant increase was found after HCG treatment. CONCLUSION: Luteal rescue with HCG is associated with a second wave of angiogenesis and vascular stabilization. PMID- 11726569 TI - The effectiveness and safety of recombinant human LH to support follicular development induced by recombinant human FSH in WHO group I anovulation: evidence from a multicentre study in Spain. AB - BACKGROUND: Until recently, human menopausal gonadotrophin (HMG), a urinary extract containing a fixed combination of LH and FSH, was the only source of exogenous LH for women with hypogonadotrophic hypogonadism undergoing ovulation induction with gonadotrophins. Recombinant human LH (rLH) is now available for clinical use, providing a new treatment option but clinical data on its use are scanty. Therefore, the aim of the present study was to investigate the efficacy and safety of rLH combined with recombinant FSH (rFSH) to induce follicular development and ovulation in World Health Organization (WHO) group I anovulatory women. METHODS: We included in this multicentre study 38 hypogonadotrophic anovulatory (WHO group I) women. Patients received 150 IU/day rFSH and 75 IU/day rLH (with the possibility of dose adjustment) as a single s.c. injection for up to three cycles with a total of 84 treatment cycles. RESULTS: Sufficient follicular growth was observed in 79 (94%) out of 84 initiated cycles. The 75 IU rLH dose was found to be effective in most treatment cycles (94%) and only five cycles in three patients required daily dose increase. Overall, HCG was administered to trigger ovulation in 67 (80%) of the 84 cycles while it was withheld in 12 cycles (14%) due to ovarian hyper-response and five cycles (6%) were cancelled for insufficient follicular growth. The pregnancy rate per started treatment cycle and per cycle given HCG was 18 and 22.4% respectively. Pregnancy was achieved by 15 (39.5%) of the 38 patients. Mild to moderate ovarian hyperstimulation syndrome occurred in three patients. Local tolerance was good. CONCLUSIONS: This study confirms that combined rFSH and rLH treatment induces follicular growth, ovulation and pregnancy in a good proportion of hypogonadotrophic anovulatory patients and is well tolerated. The doses of 150 IU rFSH and 75 IU rLH daily seem the most appropriate but in a small minority of patients doses >75 IU rLH/day may be necessary. PMID- 11726570 TI - Human ovarian steroid secretion in vivo: effects of GnRH agonist versus antagonist (cetrorelix). AB - BACKGROUND: In order to investigate whether gonadotrophin-releasing hormone (GnRH) antagonists exert a significant effect on steroid secretion in vivo compared with GnRH agonists, concentrations of sex steroid hormones (oestradiol, progesterone and testosterone) were studied in follicular fluid from women undergoing ovarian stimulation and treated with either GnRH agonist or antagonist. In addition, the correlation between follicular fluid steroid hormone concentrations and variables of follicular and oocyte development was evaluated. METHODS: Microparticle enzyme immunoassay and radioimmunoassays were used. RESULTS: The mean (SEM) follicular fluid oestradiol concentration was significantly lower in patients treated with GnRH antagonist than in those treated with GnRH agonist (542.0 +/- 76.9 versus 873.0 +/- 105.1 pg/ml, P = 0.02), which correlates with the mean serum oestradiol concentrations found in these two groups. No significant differences were found between groups in follicular fluid progesterone concentrations. Women undergoing GnRH antagonist treatment showed similar concentrations of follicular fluid testosterone compared with GnRH agonist-treated women (14.8 +/- 1.1 versus 13.3 +/- 2.7 ng/ml). The oestradiol:testosterone ratio was markedly reduced in women treated with GnRH antagonist (49.1 +/- 2.3 versus 60.1 +/- 4.4, P = 0.04). In contrast, no differences were found either in the progesterone:testosterone ratio, or in the oestradiol:progesterone ratio. CONCLUSIONS: GnRH antagonist therapy in women undergoing ovarian stimulation had a significant effect on ovarian follicular steroidogenesis. PMID- 11726571 TI - Plasma prolactin/oestradiol ratio at 38 weeks gestation predicts the duration of lactational amenorrhoea. AB - BACKGROUND: Fully breastfeeding women experience an amenorrhoea of variable duration. Our aim was to identify in pregnancy, endocrine markers that could predict the duration of subsequent lactational amenorrhoea. METHODS: We studied 17 healthy women at 34 and 38 weeks gestation, and 1 and 3 months post-partum. The women fully breastfed until 6 months post-partum. During pregnancy, prolactin (PRL), oestrogens (total oestradiol, unconjugated oestrone, unconjugated oestriol), sex hormone binding globulin (SHBG), dehydroepiandrosterone sulphate (DHEA-S), progesterone and placental lactogen, and during post-partum PRL, oestrogens and SHBG, were measured. Free oestradiol in pregnancy and post-partum was calculated. RESULTS: Ten women experienced long (>6 months) and seven experienced short (<6 months) lactational amenorrhoea. At 38 weeks gestation, the women who experienced a long lactational amenorrhoea had twice as much PRL, about half the total oestradiol, lower SHBG concentration (P < 0.05, Student's t-test, Bonferroni modification) and similar free oestradiol concentration, compared with those who experienced short lactational amenorrhoea. The difference in PRL concentration persisted in post-partum postsuckling samples. CONCLUSION: At 38 weeks gestation, the ratio PRL/oestradiol identified all individual women according to the subsequent duration of their lactational amenorrhoea, suggesting that duration of lactational amenorrhoea is conditioned during pregnancy. PMID- 11726572 TI - Ovarian steroidogenic response to human chorionic gonadotrophin in obese women with polycystic ovary syndrome: effect of metformin. AB - BACKGROUND: The aim of the present study was to investigate the steroidogenic response pattern to HCG in obese women with polycystic ovary syndrome (PCOS) and the possible effects of metformin treatment on it. METHODS: A single injection of human chorionic gonadotrophin (HCG, 5000 IU) was given to 12 obese [body mass index (BMI) > or = 27 kg/m2] women with PCOS and to 27 control women. Blood samples for assays of 17alpha-hydroxyprogesterone (17-OHP), androstenedione, testosterone and oestradiol were collected at baseline and 1, 2 and 4 days after the injection. Responses to HCG were also assessed in the PCOS women after 2 month treatment with metformin (500 mg x 3 daily). RESULTS: Serum 17-OHP and oestradiol concentrations peaked at 24 h in the PCOS women and preceded the maximum testosterone concentration, which was seen at 48 h. In the control women the maximum concentrations of all these steroids were reached 96 h after HCG. After metformin treatment, the basal serum testosterone concentration and the areas under the androstenedione (AUC(A)) and testosterone (AUC(T)) response curves after HCG decreased significantly. CONCLUSIONS: The results demonstrate that obese PCOS women have a male-type steroidogenic response pattern to a single injection of HCG and a higher androgen secretory capacity than control women, which may be explained by the increased thecal cell activity in the polycystic ovary. The slight alleviation of hyperandrogenism brought about by metformin therapy appears to be due to its effect on ovarian steroidogenesis possibly mediated by decreased insulin action. PMID- 11726573 TI - Follistatin and activin A serum concentrations in obese and non-obese patients with polycystic ovary syndrome. AB - BACKGROUND: Activin promotes ovarian follicular development, inhibits androgen production and increases FSH and insulin secretion. Follistatin, an activin binding protein, neutralizes activin bioactivity. Therefore, a decrease in the ratio of activin/follistatin might encourage characteristic features of polycystic ovary syndrome (PCOS). We investigated whether women with PCOS showed disordered follistatin and/or activin serum concentrations. METHODS: The study group included 24 obese and 20 non-obese (body mass index vertical line and <27 kg/m2 respectively) clomiphene-failure PCOS patients. The control group included 16 obese and 46 non-obese patients with normal ovulatory cycles. Blood samples were obtained from the patients on day 3-5 of a progesterone-induced or spontaneous cycle and were assayed for LH, FSH, testosterone, 17-hydroxy progesterone, androstenedione, follistatin, activin A, fasting glucose and insulin. RESULTS: Follistatin concentrations were comparable between obese and non-obese PCOS patients (mean +/- SE; 1171 +/- 103 and 1045 +/- 159 pg/ml respectively) and significantly higher than their respective controls (628 +/- 61 and 592 +/- 49 pg/ml, P < 0.0001 and P < 0.02 respectively). Activin A concentrations were comparable between the four groups (590 +/- 35, 513 +/- 74, 661 +/- 87 and 595 +/- 43 pg/ml in obese and non-obese PCOS and controls respectively). Stepwise regression analyses for relationships between follistatin or activin A levels and all other variables indicated that follistatin was significantly and independently positively affected by PCOS (P < 0.0001), age (P < 0.02), androstenedione (P < 0.03) and weight (P < 0.05). Activin A was significantly and independently negatively affected by PCOS (P < 0.003) and FSH (P < 0.03), and positively affected by weight (P < 0.009) and androstenedione (P < 0.02). CONCLUSIONS: Serum follistatin is increased in PCOS patients, regardless of obesity. PCOS is the most significant variable that relates to high follistatin and low activin A serum concentration. A high follistatin/activin ratio could well contribute to the pathophysiology of PCOS. PMID- 11726574 TI - The cost-effectiveness of IVF in the UK: a comparison of three gonadotrophin treatments. AB - BACKGROUND: The objective of this study was to evaluate the cost-effectiveness of women undergoing IVF treatment with recombinant FSH (rFSH) in comparison with highly purified urinary FSH (uFSH-HP) and human menopausal gonadotrophins (HMG). METHODS: A decision-analytic model was used to estimate cost-effectiveness ratios for 'the average cost per ongoing pregnancy' and 'incremental cost per additional pregnancy' for women entering into IVF treatment for a maximum of three cycles. The model was constructed based on a previously published large prospective randomized clinical trial comparing rFSH and uFSH-HP. Where necessary, these data were augmented with a combination of expert opinion, evidence from the literature and observational data relating to the management and cost of IVF treatment in the UK. The cost of rFSH, uFSH-HP and HMG were obtained from National Health Service list prices in the UK. RESULTS: The model predicted a cumulative pregnancy rate after three cycles of 57.1% for rFSH and 44.4% for both uFSH-HP and HMG. The cost of IVF treatment was 5135 pounds sterling for rFSH, 4806 pounds sterling for uFSH-HP and 4202 pounds sterling for HMG. When assessed in association with outcomes, the average cost per ongoing pregnancy was more favourable with rFSH (8992 pounds sterling) than with either uFSH-HP (10 834 pounds sterling) or HMG (9472 pounds sterling). The incremental cost per additional pregnancy was 2583 pounds sterling using rFSH instead of uFSH-HP and 7321 pounds sterling using rFSH instead of HMG. These results were robust to changes in the baseline assumptions of the model. CONCLUSION: rFSH is a cost effective treatment strategy in ovulation induction prior to IVF. PMID- 11726575 TI - Cost-effectiveness modelling of recombinant FSH versus urinary FSH in assisted reproduction techniques in the UK. AB - BACKGROUND: The purpose of this study was to undertake an economic evaluation to compare the cost-effectiveness of recombinant (r)FSH with urinary (u)FSH for attaining clinical pregnancy with assisted reproduction. METHODS: Mathematical modelling was utilized incorporating a Markovian decision framework and a Monte Carlo simulation. Statistical representations of recurrent events over time were incorporated into a decision analysis involving fresh and frozen cycles in any sequence (after the first fresh embryo transfer cycle) over three successive assisted reproduction attempts. The mean values of transition probabilities were derived from randomized controlled clinical trials and published reports. The distributions of these transition probabilities were agreed upon by a panel of experts. Cost data for procedures and drugs were derived and validated according to the perspectives of the National Health Service and private clinics in the UK. RESULTS: The study involved 5000 Monte-Carlo simulations of treatment on a Markov cohort of 100 000 patients. The total number of pregnancies attained was significantly higher in the rFSH (40 575) compared with the uFSH (37 358) group. The cost per successful pregnancy was significantly lower for rFSH (5906 pounds sterling) compared with uFSH (6060 pounds sterling) and overall, fewer cycles of treatment were required with rFSH to achieve an ongoing pregnancy. The incremental cost-effectiveness ratio is 4148 pounds sterling for each additional clinical pregnancy with rFSH. CONCLUSIONS: In addition to the increased effectiveness of rFSH in ART, this study demonstrated that it is more cost effective and more efficient than uFSH in attaining an ongoing pregnancy. PMID- 11726577 TI - Transvaginal ultrasound-guided embryo transfer improves pregnancy and implantation rates after IVF. AB - BACKGROUND: Attempts are constantly being made to improve clinical pregnancy rates after IVF and embryo transfer. Since November 1998, we have gradually been adopting transvaginal ultrasound guidance during embryo transfer. We retrospectively examined the efficacy of this method on pregnancy and implantation rates. METHODS: The results of 846 cycles from our IVF-embryo transfer programme were analysed and comparisons were made between those carried out using ultrasound guidance and those by the clinical touch method. RESULTS: Higher pregnancy and implantation rates (28.9 and 15.2% respectively) were found in the group using the transvaginal ultrasound guidance during embryo transfer compared with those in the group using the clinical touch method (13.1 and 7.0% respectively). The differences were statistically significant (P < 0.01). There was no significant difference in ectopic pregnancy rates between the two groups. CONCLUSION: The use of transvaginal ultrasound-guided embryo transfer significantly improved both pregnancy and implantation rates. Although technically difficult, we suggest its use may maximize the chances of achieving a successful pregnancy outcome. PMID- 11726576 TI - Suppression of spermatogenesis to azoospermia by combined administration of GnRH antagonist and 19-nortestosterone cannot be maintained by this non-aromatizable androgen alone. AB - BACKGROUND: For male hormonal contraception, combined administration of gonadotrophin-releasing hormone (GnRH) antagonists and androgens effectively suppresses spermatogenesis to azoospermia. In non-human primates this suppression can be maintained more easily by androgens alone. METHODS: A clinical trial with six healthy volunteers was performed to test this approach in man. Loading doses of 10 mg/day of the GnRH antagonist cetrorelix were given subcutaneously for 5 days, followed by maintenance doses of 2 mg/day up to week 12. At 2 weeks after the first GnRH antagonist injection, androgen substitution was initiated with a loading dose of 400 mg 19-nortestosterone hexyloxyphenylpropionate (19NT-HPP) intramuscularly, followed by injections of 200 mg 19NT-HPP every 3 weeks up to week 26. RESULTS: Serum concentrations of LH, FSH and testosterone were effectively suppressed by cetrorelix administration. Within 12 weeks, azoospermia was achieved in all six volunteers. After cessation of cetrorelix injections in week 12, gonadotrophins and testosterone increased significantly despite continued 19NT-HPP injections. In parallel, spermatogenesis was restimulated in five of six volunteers. CONCLUSIONS: Combined administration of cetrorelix and 19NT-HPP leads to azoospermia within 3 months. However, complete azoospermia cannot be maintained by continued injections of the non-aromatizable 19NT-HPP alone. PMID- 11726578 TI - Ovarian response during IVF-embryo transfer cycles after laparoscopic ovarian cystectomy for endometriotic cysts of >3 cm in diameter. AB - BACKGROUND: Ovarian response during IVF cycles after laparoscopic ovarian cystectomy for endometriotic cysts >3 cm is controversial. A retrospective study was designed to study this problem. METHODS: At laparoscopy, endometriomas >3 cm were treated by ovarian cystectomy, whilst adhesions and peritoneal endometriosis were treated using conventional techniques. Ovarian stimulation was achieved with clomiphene and gonadotrophins or with gonadotrophins after a desensitization with gonadotrophin-releasing hormone agonists. Three groups of patients were retrospectively selected from an IVF-embryo transfer database: patients who underwent laparoscopic ovarian cystectomy for an endometrioma >3 cm (Group A, n = 41), patients with endometriosis without ovarian endometrioma (Group B, n = 139) and patients with tubal infertility (Group C, n = 59). RESULTS: The groups did not differ in age. In the first IVF cycle, the mean (+/- SD) numbers of oocytes and of embryos were 9.4 +/- 6.2 and 4.7 +/- 3.6 respectively in group A and 11.6 +/- 7.5 and 5.1 +/- 4.9 in group B (not significant). The results did not differ in cycles 2 and 3 or when compared according to age. No difference was found when comparing patients with endometriosis and patients with tubal infertility. CONCLUSION: The number of oocytes and embryos obtained was not significantly decreased by laparoscopic cystectomy, suggesting that in experienced hands this procedure may be a valuable surgical tool for the treatment of large ovarian endometriomas. However, great care must be taken to avoid ovarian damage. PMID- 11726579 TI - Natural cycle IVF in unexplained, endometriosis-associated and tubal factor infertility. AB - BACKGROUND: To elucidate possible differences between unexplained and minimal peritoneal endometriosis-associated infertility, we studied their outcome in natural cycle IVF (NIVF). METHODS: A prospective cohort study was carried out on unexplained (33 couples), minimal peritoneal endometriosis-associated (30 couples) and tubal factor (24 couples) infertility in 223 NIVF cycles, using human chorionic gonadotrophin (HCG) for ovulation induction. RESULTS: During the first NIVF attempt, follicular and luteal phase oestradiol, FSH, LH and progesterone concentrations, as well as endometrial thickness and follicular diameter were similar among the three groups. Periovulatory follicular growth monitored from day of HCG administration to oocyte aspiration was significantly lowered in unexplained infertility compared with minimal endometriosis-associated and tubal factor infertility. The fertilization rate, clinical pregnancy rate per initiated cycle, per successful oocyte retrieval and per embryo transfer, in minimal endometriosis (80.0, 10.4, 16.0 and 23.5% respectively) were similar to that in tubal factor infertility patients (68.6, 5.8, 11.4 and 16.0%) but significantly higher (P < 0.05) than that of the unexplained infertility group (62.2, 2.6, 5.4 and 8.7%). CONCLUSIONS: The significant reduction in follicular periovulatory growth, fertilization and pregnancy rates in unexplained infertility compared with minimal peritoneal endometriosis patients may be explained by sub-optimal follicular development with possibly reduced oocyte quality, intrinsic embryo quality factors or by impaired implantation. From a clinical point of view, NIVF is less suited to unexplained infertility treatment, but might represent an interesting treatment option for minimal peritoneal endometriosis-associated infertility. PMID- 11726580 TI - Review of unexplained infertility and obstetric outcome: a 10 year review. AB - BACKGROUND: Increased maternal and fetal risks have been reported in pregnancies following unexplained infertility. Our aims were to examine the obstetric and perinatal outcome of singleton pregnancies in couples with unexplained infertility and explore the impact of fertility treatment. METHODS: Women with unexplained infertility were identified from the Aberdeen Fertility Clinic Database. Their unit numbers were matched against the Aberdeen Maternity and Neonatal Databank (AMND) in order to extract obstetric records of those women with subsequent pregnancy outcomes. The general obstetric population served as a control group. RESULTS: Women with unexplained infertility were older [30.8 versus 27.9 years, 95% confidence interval (CI) for difference = +2.4 to +3.4] and more likely to be primiparous (59 versus 40%, 95% CI = +1.3 to +1.9). After adjusting for age and parity they had a higher incidence of pre-eclampsia, abruptio placentae, preterm labour, emergency Caesarean section and induction of labour in comparison with the general population (P < 0.05). Perinatal outcome did not differ between women with unexplained infertility and those of the general population. The multiple pregnancy rate was 5.4% higher following fertility treatment than in women who conceived spontaneously (95% CI = +2.8 to +9.7). CONCLUSIONS: Women with unexplained infertility are at higher risk of obstetric complications which persist even after adjusting for age, parity and fertility treatment. The reasons are however unclear and merit further study. PMID- 11726581 TI - Trends in the incidence of births and multiple births and the factors that determine the probability of multiple birth after IVF treatment. AB - BACKGROUND: We aimed to review trends in the probability of birth and multiple birth before and after the legal restriction limiting the maximum allowable number of embryos transferred, and to examine factors that determine the probability of multiple birth following IVF treatment. METHODS: We analysed data relating to 7170 IVF and 530 intracytoplasmic sperm injection cycles reaching embryo transfer undertaken by 4417 women at a single tertiary referral assisted conception centre in the UK between 1984 and 1997. Probability of birth, and of proportion of multiple births among those who gave birth, was explored using logistic regression analysis. RESULTS: Between 1984 and 1997 there was a significant increase in probability of birth but no change in the probability of multiple birth. The trend in probability of birth was almost wholly explained by the significant increase in number of embryos created per cycle. Pooling all the data, risk factors for increased chance of birth and multiple birth were: younger age (<35 years), diagnoses other than tubal infertility, fewer than three previous unsuccessful cycles, previous IVF live birth and a large number of embryos created. Given these factors, increasing the number of embryos transferred did not increase the chance of a birth, but did increase the chance of a multiple birth. CONCLUSIONS: The probability of birth has increased and the probability of multiple birth has remained unchanged, despite legislation limiting the number of embryos transferred in the UK. Efforts should be made to reduce the incidence of multiple births by transferring fewer embryos, especially in the presence of good prognostic factors for birth. PMID- 11726582 TI - Polycystic ovarian syndrome and the risk of spontaneous abortion following assisted reproductive technology treatment. AB - BACKGROUND: A high proportion of infertile patients have polycystic ovarian syndrome (PCOS) with a reportedly greater risk of spontaneous abortion. Because of the close link between PCOS and obesity and the independent association of obesity with poor pregnancy outcomes, it is important to distinguish the possible confounding effect of body mass index (BMI) or other variables from that of PCOS. This study aims to determine the effect of PCOS status on the risk of spontaneous abortion with adjustment for body mass and several other confounding factors in a large cohort of pregnant infertile women. METHODS: The patients (n = 1018) were treated in a tertiary infertility centre. Their PCOS status was determined by standard criteria and their BMI had been taken less than 1 year before the pregnancy. Patients whose PCOS status or BMI measurements were not assessed were excluded. Student's t-test or chi2 test were used to test the difference between the PCOS and non-PCOS groups while a multivariate logistical regression model was used to assess the effect of PCOS, BMI and other confounding factors. RESULTS: Overall, the incidence of PCOS was 37% in this cohort. The overall incidence of spontaneous abortion in the study population was 21%. Univariate analysis showed that women with PCOS had a significantly greater risk of spontaneous abortion compared with non-PCOS women (25 versus 18%, P < 0.01). However, using multivariate logistic regression analysis this effect was reduced to a non significant level [odds ratio (OR) = 1.10, 95% confidence interval (CI) 0.85 1.36] after adjusting for obesity and patients/treatment combination factor, and to nil after adjusting for all confounding factors considered in this study (OR = 0.98, 95% CI 0.75-1.28). CONCLUSION: The results of this study suggest that the higher risk of spontaneous abortion observed in women with PCOS is likely to be due to their high prevalence of obesity and the type of treatment they receive. PMID- 11726583 TI - Endometrial cavity fluid is associated with poor ovarian response and increased cancellation rates in ART cycles. AB - BACKGROUND: Endometrial cavity fluid (ECF) is occasionally observed during assisted reproductive technology (ART) cycles. However, few reports have described its prevalence or significance. METHODS AND RESULTS: We examined the relationships between ECF, clinical pregnancy rate (CPR), tubal factor infertility and ultrasound-visible (USV) hydrosalpinges. In 843 ART cycles involving 721 patients, ECF was observed during stimulation in 57 cycles and after human chorionic gonadotrophin (HCG) administration in 12 cycles, with an overall incidence of 8.2% (69/843). When ECF was observed during stimulation, the cancellation rate due to poor ovarian response was significantly higher (29.8 versus 16.9%, P <0.05) and the CPR per started cycle was significantly lower (26.3 versus 42.4%, P <0.05) than cycles without ECF. When ECF developed after HCG administration, the CPR was similar compared with that of the group for which ECF was not observed. In the 327 cycles involving tubal factor infertility patients, USV hydrosalpinges were noted in 71 cycles (71/327; 21.7%), and ECF developed in five of those cycles (5/71; 7.0%). A total of 27 cycles during which ECF developed (27/57, 47.4%) involved non-tubal factor patients. CONCLUSIONS: ECF during stimulation was associated with increased cancellation rates and lower CPRs per started cycle, and was not associated with USV hydrosalpinges. Furthermore, ECF observed after HCG administration did not impact CPR and may represent a different clinical entity. PMID- 11726584 TI - Outcome of ICSI using fresh and cryopreserved-thawed testicular spermatozoa in patients with non-mosaic Klinefelter's syndrome. AB - BACKGROUND: Recently, intracytoplasmic sperm injection (ICSI) of testicular spermatozoa retrieved surgically from patients with non-mosaic Klinefelter's syndrome resulted in several deliveries. The aim of this study was to evaluate the outcome of ICSI using fresh and cryopreserved-thawed testicular spermatozoa in these patients. METHODS AND RESULTS: Following informed consent regarding the genetic risks of their potential offspring, mature testicular spermatozoa were found in five out of 12 (42%) patients who underwent testicular sperm extraction, and ICSI was performed while excess tissue was cryopreserved. The mean age of the patients was 28.7 +/- 3.6 (range 23-36 years). Their baseline FSH was elevated (mean 38.3 +/- 11.4; range 22-58 mIU/ml). All patients had small testicles of 2-4 ml in volume. The outcome of ICSI using fresh or cryopreserved-thawed testicular spermatozoa during five cycles in each group, was compared. No statistical significant difference was found in the two pronuclear fertilization rate (66 versus 58%), embryo cleavage rate (98 versus 90%) and embryo implantation rate (33.3 versus 21.4%) for fresh or cryopreserved sperm accordingly. The clinical outcome after using fresh testicular sperm included two singleton pregnancies (one delivered and one ongoing) and a triplet pregnancy resulting in a twin delivery (after reduction of an 47,XXY embryo). After using cryopreserved-thawed testicular spermatozoa, two pregnancies were obtained resulting in one delivery of twins and one early spontaneous abortion. CONCLUSIONS: Outcome of ICSI using cryopreserved-thawed testicular spermatozoa of patients with non-mosaic Klinefelter's syndrome is comparable with that following the use of fresh spermatozoa. The genetic implications for the future offspring should be explained to the patients. PMID- 11726585 TI - Serial ultrasonography, hormonal profile and antisperm antibody response after testicular sperm aspiration. AB - BACKGROUND: In many fertility centres, intracytoplasmic sperm injection (ICSI) with epididymal or testicular spermatozoa is a routine treatment for men with azoospermia. In this prospective study, the physiological consequences after testicular sperm aspiration (TESA), using suction and a 19 gauge needle, were evaluated. METHODS AND RESULTS: Thirty-five consecutive men with azoospermia underwent TESA. Testicular ultrasonography with Doppler flow imaging was performed and testicular volumes were evaluated pre-operatively and 3 months after aspiration. If focal testicular lesions were found, further examinations were performed 6 and 9 months after TESA. Serum FSH, testosterone and antisperm antibodies (ASA) were analysed. Focal testicular lesions were seen in four out of 61 testes (6.6%) at the 3 month investigation point. Three lesions were resolved after 6 months and all after 9 months. Testicular echogenicity remained unchanged in 50 cases (82%) 3 months after TESA. Four men (11.4%) reported severe subjective discomfort post-operatively, but only one had a medical consultation where an intratesticular haematoma was diagnosed. There were no significant changes in FSH and testosterone after surgery and testicular volumes were similar after 3 months. There were three borderline cases of ASA in serum, but none was classified as ASA-positive. CONCLUSIONS: The puncture method of testicular sperm aspiration seems to be a safe method for sperm retrieval, with minimal physiological consequences. PMID- 11726586 TI - Laser assisted immobilization of spermatozoa prior to intracytoplasmic sperm injection in humans. AB - BACKGROUND: The conventional method of immobilization of spermatozoa prior to intracytoplasmic sperm injection (ICSI) is mechanical breakage of the tail by pressing it against the bottom of the injection dish. METHODS: This prospective self-controlled study was set up to evaluate the potential of a non-contact 1.48 microm wavelength diode laser in terms of immobilization. In addition, the fertilization rate and further development potential of such zygotes were investigated. The patients included in our study (n = 60) had oestradiol concentrations >2000 pg/ml, and thus a relatively high number of MII oocytes could be expected. Approximately half the oocytes were injected with laser treated spermatozoa (n = 262, study group) and the other half with mechanically immobilized spermatozoa (n = 252, control group). RESULTS: No significant differences between the two groups in terms of fertilization rate, early cleavage or blastocyst formation were observed. However, time required for identification, aspiration and injection of a potential spermatozoa was significantly shorter in the laser immobilized sperm group (P < 0.001). CONCLUSIONS: The application of a non-contact diode laser for sperm immobilization prior to ICSI is a potentially useful alternative to the conventional mechanical approach. PMID- 11726587 TI - Long-term outcomes of elective human sperm cryostorage. AB - BACKGROUND: Sperm cryopreservation allows men with threatened fertility to preserve their progenitive potential, but there is little data on long-term outcomes of elective sperm cryostorage programmes. METHODS AND RESULTS: Over 22 years, 930 men sought semen cryostorage in a single academic hospital, of which 833 (90%) had spermatozoa cryostored. Among 692 (74%) men surviving their illness, sperm samples were discarded for 193 (21% of all applicants, 28% of survivors) and cryostored spermatozoa were used for 64 men (7% of all applicants, 9% of survivors) in 85 treatment cycles commencing at a median of 36 months post storage (range 12-180 months) with nearly 90% of usage started within 10 years of storage and none after 15 years. Pregnancy was most efficiently produced by intracytoplasmic sperm injection (median three cycles) compared with conventional IVF (median eight cycles) or artificial insemination (median more than six cycles; P < 0.05). A total of 141 (15%) of men had died and of these, 120 (85% of those dying) had their spermatozoa discarded; requests to prolong cryostorage were received from relatives of 21 men (2% of all applicants, 15% of deceased) of which three cases had spermatozoa transferred for use with no pregnancies reported. Sperm concentration was lower for all cryostorage groups compared with healthy sperm donor controls (P < 0.05). Following orchidectomy, men with testicular cancer had sperm density approximately half that of all other groups of men seeking cryostorage (P < 0.05), the lowering attributable to removal of one testis rather than in defects in spermatogenesis. CONCLUSION: Elective sperm cryopreservation is an effective, if sparsely used, form of fertility insurance for men whose fertility is threatened by medical treatment and is an essential part of any comprehensive cancer care programme. PMID- 11726588 TI - Assisted reproduction with in-vitro-cultured testicular spermatozoa in cases of severe germ cell apoptosis: a pilot study. AB - BACKGROUND: Apoptosis-related cell damage is known to compromise success rates of assisted reproduction with ejaculated spermatozoa. This study was undertaken to determine whether the frequency of apoptosis-related cell damage and reproductive performance of testicular spermatozoa from men with non-obstructive azoospermia can be improved by in-vitro culture. METHODS: Testicular tissue samples were cultured for 2 days in the presence of 50 IU/l FSH and 1 micromol/l testosterone. The frequency of spermatozoa showing DNA strand breakage and plasma membrane phosphatidylserine externalization was compared in before-culture and after culture samples. The after-culture samples were used in assisted reproduction attempts. RESULTS: In a group of 11 azoospermic patients with at least two previous intracytoplasmic sperm injection (ICSI) failures, the incidence of DNA strand breakage was high in living testicular spermatozoa from before-culture samples, but significantly lower in after-culture samples (96 versus 30%, P < 0.001). The same applied to the incidence of phosphatidylserine externalization in the motile sperm subpopulation from the before-culture and after-culture samples (83 versus 6%, P < 0.001). Seven ongoing clinical pregnancies (six with fresh embryos and one with cryopreserved embryos) were established. CONCLUSIONS: Severe testicular sperm apoptosis may become a new indication for testicular tissue in-vitro culture before ICSI. PMID- 11726589 TI - Familial sperm polyploidy induced by genetic spermatogenesis failure: case report. AB - We report a case of oligoasthenoteratozoospermia in a 40 year-old patient with a familial history that revealed multiple cases of infertility and perinatal deaths. The patient's semen sample contained 2x10(6) spermatozoa/ml, with an overall progressively motile population of <5%. Cytological analysis revealed a teratozoospermia with 100% of abnormal macrocephalic sperm heads and an irregular acrosomal cap in 38% of cells. Moreover, 72% of spermatozoa carried multiple flagella (2-5). The midpiece was elongated and/or enlarged with cytoplasmic droplets in 15% of cells. The multiple anomalies index (MAI) was 3.3 (normal value = 1.6), reflecting the high incidence of spermatozoal morphological abnormalities in this patient. Ultrastructural analysis revealed the presence of 2 or 3 vacuolated nuclei per sperm head. The acrosome was abnormal and the chromatin, partially packaged, appeared rough. In some cases, a large amount of cytoplasm containing vacuoles was observed around the nucleus and the acrosome. The mitochondrial helix was disorganized. Chromosome analysis performed on blood cells revealed a normal karyotype. Three-colour fluorescence in-situ hybridization (FISH) analysis of 1148 spermatozoa showed 21.6% to be diploid, 62.4% triploid, 13.3% quadriploid and 2.7% hyperploid (<4n). In conclusion, we suggest that this case could result from a genetically induced spermiation failure, the origin of which is discussed. PMID- 11726590 TI - Early embryo cleavage is a strong indicator of embryo quality in human IVF. AB - BACKGROUND: In order to decrease multiple birth rates without decreasing birth rates overall, it is important to increase the capability of selecting the most optimal embryos for transfer. It has been shown that human embryos which cleave early, i.e. complete the first mitotic division within 25-27 h post insemination, provide higher pregnancy and implantation rates. METHODS AND RESULTS: In this prospective study, an evaluation of 10 798 scored embryos showed that early cleavage resulted in a significantly higher proportion of good quality embryos compared with late cleavage (62.5 versus 33.4%, P < 0.0001). When examining both day 2 and day 3 transfers together, early-cleaving embryos (306 transfers) gave rise to significantly higher rates of pregnancy/transfer (40.5 versus 31.3%, P = 0.0049), implantation (28.0 versus 19.5%, P = 0.0001) and birth/ongoing pregnancy (34.3 versus 24.0%, P = 0.0009) than did late-cleaving embryos (521 transfers). A stepwise logistic regression of all data showed that the total number of good quality embryos and female age were independent predictors of both pregnancies and birth. For intracytoplasmic sperm injection (ICSI) embryos, early cleavage was found to be an independent predictor of birth. CONCLUSIONS: Early embryo cleavage is a strong biological indicator of embryo potential, and may be used as an additional embryo selection factor for ICSI embryos. PMID- 11726591 TI - Early cleavage of human embryos: an effective method for predicting successful IVF/ICSI outcome. AB - BACKGROUND: The need for effective parameters for selecting the best embryos is paramount when a large number of them are available for transfer. Other studies have reported that transfer of pre-selected embryos, based on cleavage to the 2 cell stage at 25 h and 27 h post-insemination/intracytoplasmic sperm injection (ICSI), increases implantation and pregnancy rates. We investigated whether extending the time for selection of cleaved embryos to 29 h post insemination/ICSI had a similar effect on pregnancy and implantation rates. METHODS: Cleavage to the 2-cell stage was assessed at 25, 27 and 29 h post insemination/ICSI. Embryos that had cleaved at any of these time points were designated as 'early cleavage' (EC), while others were designated as 'non-early cleavage' (NEC). EC embryos were selected and preferentially transferred. RESULTS: EC occurred in 57% of the cycles (61% IVF; 51% ICSI). Significantly (P = 0.02) more clinical pregnancies occurred in the EC group (23/42, 55%) compared with the group that had no embryo undergoing first cleavage up to 29 h post insemination/ICSI (8/32, 25%). The EC group of patients was significantly younger than the NEC. CONCLUSION: Transfer of selected embryos that reached the 2-cell stage between 25 and 29 h post-insemination/ICSI is a reliable prognostic tool for patients undergoing assisted reproduction techniques. PMID- 11726592 TI - The mechanism of hydrosalpinx in embryo implantation. AB - BACKGROUND: Hydrosalpinx adversely affects embryo implantation and contributes to poor implantation rates post embryo transfer. Embryo transport depends on concomitant intrauterine fluid motion induced by uterine wall motility, the result of spontaneous myometrial contractions towards the fundus. METHODS AND RESULTS: The uterine dynamics of five patients with hydrosalpinx were recorded and analysed by image-processing techniques: the frequency was higher while the amplitudes and passive widths were lower compared with healthy volunteers. The existing peristaltic activity should have induced intrauterine fluid flow; however, the recordings failed to show the expected transport of fluid bolus. This observation was supported by mathematical simulations based on the hypothesis that fluid accumulation in the Fallopian tube of a patient with hydrosalpinx increases tubal pressure and thereby induces a pressure gradient between the fundus and the cervix. This pressure gradient acts adversely to the cervix-to-fundus intrauterine peristalsis and generates reflux currents that may thrust embryos away from the implantation site. CONCLUSIONS: The reflux phenomenon could explain the reduced implantation rate associated with hydrosalpinx. Resolution of the issue of whether the removal of a Fallopian tube with hydrosalpinx should be undertaken for improving IVF pregnancy rates should be accompanied by prospective randomized clinical trials. PMID- 11726593 TI - Relationship between stage, site and morphological characteristics of pelvic endometriosis and pain. AB - BACKGROUND: The relationship between frequency and severity of pain symptoms and site, stage and morphological characteristics of endometriotic lesions was analysed in a multicentre cross-sectional observational study. METHODS: A total of 469 women (median age 31 years, range 18-45) who met the following criteria were consecutively observed in the participating centres during the study period: age 18-45 years, first laparoscopic or laparotomic diagnosis of endometriosis, pain symptoms lasting > or = 6 months, pain as the main or only complaint of the condition, absence of pelvic anomalies and no previous pelvic surgery. Dysmenorrhoea and non-menstrual pain were evaluated using a multidimensional verbal rating scale. The women were requested to grade the severity of dysmenorrhoea, non-menstrual pelvic pain and deep dyspareunia using a 10-point linear analogue scale. RESULTS: Dysmenorrhoea was present in 77% of subjects with ovarian endometriosis, 88% of those with endometriosis of the peritoneum, 92% of subjects with endometriosis of both ovary and peritoneum and in all the subjects with endometriosis of rectovaginal septum. These differences were not statistically significant after Bonferroni's correction. No marked difference emerged between the severity of dysmenorrhoea and site of endometriosis, but women with ovarian endometriosis tended to have lower scores (not significant). No clear association emerged between frequency and severity of non-menstrual pain, dyspareunia and site of endometriosis and the presence and severity of dysmenorrhoea, non-menstrual pain and dyspareunia. Dyspareunia was more frequently reported in women with only atypical endometriosis (56.8%) versus 47.7% in women with typical endometriosis, but with borderline significance (P = 0.05). Dyspareunia occurred in 68.2% of patients with both typical and atypical lesions. CONCLUSIONS: The results of this study find no clear-cut association between stage, site or morphological characteristics of pelvic endometriosis and pain. PMID- 11726594 TI - Transvaginal colour Doppler in patients with ovarian endometriomas and pelvic pain. AB - BACKGROUND: The aim of this investigation was to correlate ovarian endometrioma vascularization with the presence of pelvic pain. METHODS: The presence of blood flow, peak systolic velocity (PSV, cm/s) and lowest pulsatility index (PI), assessed by transvaginal colour Doppler ultrasonography and CA-125 plasma concentrations, were retrospectively analysed in 74 patients who had undergone operations for cystic ovarian endometriosis. Fifty-two patients were asymptomatic (group A) and 22 presented with pelvic pain (group B). There were 56 endometriomas in group A and 26 in group B. RESULTS: Blood flow was found in 66.1 and 88.5% of endometriomas in groups A and B respectively (P = 0.036). PI was significantly lower (P = 0.009) and CA-125 concentration higher (P = 0.0004) in group B. There were no differences in PSV. CONCLUSIONS: We conclude that vascularization of ovarian endometriomas in patients presenting with pelvic pain is higher than in asymptomatic patients. This could be an indicator of endometriosis activity. PMID- 11726595 TI - Endometriosis: absence of recurrence in patients after endometrial ablation. AB - BACKGROUND: The present study was undertaken to evaluate differences between patients with and without eutopic endometrium in the recurrence of ectopic endometriotic implants. METHODS: Endometrial ablation (EA) was carried out in 14 women out of 28 laparoscopically treated for endometriosis and recurrence of the disease was evaluated 24 months later. Data were compared using paired Student's t-test and chi2 test. RESULTS: Patients undergoing EA procedures did not exhibit recurrence of endometriosis while nine patients without that procedure had recurrence of the disease (P < 0.001). The endometrial cells found in the debris of the cul de sac of eight patients who did not undergo EA were both stromal and epithelial cells. No blood or blood cells were found in the cul de sac of patients undergoing EA. CONCLUSIONS: The present study supports a role of eutopic endometrium in the recurrence of endometriosis through tubal dissemination of endometrial debris and implantation of endometrial cells into the abdomen. PMID- 11726596 TI - Ultrastructure of epithelial plaque formation and stromal cell transformation by post-ovulatory chorionic gonadotrophin treatment in the baboon (Papio anubis). AB - BACKGROUND: To understand factors controlling endometrial responses to pregnancy, we have established a model using the baboon and examined the effects of infused human chorionic gonadotrophin (HCG) on the preparation of the luminal epithelium and stromal cell differentiation for the establishment of pregnancy. METHODS: The ultrastructure of endometrium from normal day 10 post-ovulation animals, cycling females treated with either HCG or FSH (control), and a day 15 pregnant animal has been compared. RESULTS: In the control endometrium, the luminal epithelium was smooth and regular, with underlying spindle shaped stromal cells. In pregnancy, the luminal epithelium underwent a plaque reaction, while stromal cells enlarged and developed filament-rich cell processes. Infusion of HCG produced changes similar to those seen in pregnancy, with generalized plaque formation and stromal decidualization, while in the animal treated with FSH there was no response. CONCLUSIONS: This study indicates that infusion of HCG into the uterus can duplicate many of the responses of the endometrium to pregnancy, although in this case the plaque reaction involved the whole of the luminal epithelium, rather than only the implantation site as in pregnancy. PMID- 11726597 TI - Mortality in a cohort of IVF patients. AB - BACKGROUND: Risks associated with IVF and related assisted reproduction technologies include complications of ovarian stimulation, surgical procedures and pregnancy itself. Serious complications are uncommon but may be potentially life threatening. The aims of this study were to compare the mortality rates of women who received IVF treatment, as well as those who were referred but were not treated, with the mortality rate in the general female population, to determine the maternal mortality rate following IVF conception and to establish whether any deaths had occurred as a result of treatment complications. METHODS: Deaths were identified in a cohort of 29 700 Australian IVF patients by record-linkage with the National Death Index and a cancer registry. RESULTS: The all-cause mortality rates in IVF patients (treated and untreated) were significantly lower than in the general female population of the same age. In treated women, 72 deaths were observed and 125 deaths were expected giving an age-standardized mortality ratio of 0.58 (95% confidence interval 0.48-0.69). Two maternal deaths were identified in the 42 days of the puerperium. Complications of ovarian hyperstimulation syndrome could not be directly related to any of the deaths identified in this cohort. CONCLUSIONS: As well as providing some reassurance about the safety of IVF treatments, the findings point to the existence of a 'healthy patient effect' whereby the unhealthiest women in the population are deterred from pregnancy and infertility treatment. PMID- 11726598 TI - Regional differences in waiting time to pregnancy among fertile couples from four European cities. AB - BACKGROUND: A previous European study found a longer time to pregnancy (TTP) among fertile women from Paris compared with women from other Western European countries. A co-ordinated, cross-sectional study of pregnant couples from Denmark (Copenhagen), France (Paris), Scotland (Edinburgh) and Finland (Turku) was therefore undertaken to assess differences in waiting TTP among couples from these cities. METHODS: Pregnant women were invited to participate when they showed up for their first antenatal visit in one of the four centres. Inclusion criteria included that their partner was 20-45 years of age and born in the country in which he was currently living and that the pregnancy was achieved without fertility treatment. Both partners filled in a questionnaire and the man underwent a physical examination and delivered a semen sample (Turku: n = 237, Copenhagen: n = 302, Edinburgh: n = 212, Paris: n = 191). RESULTS: French couples had a decreased probability of conception compared with couples from the other three countries, although only after adjustment for confounders. No significant differences between couples from the three other countries were found. CONCLUSION: The observed geographical differences in TTP remain unexplained and were not due to differences in semen quality, but may be caused by varying exposures to environmental factors or psychological distress. In addition, selection bias due to the low participation rates cannot be ruled out. Future studies examining the causes for geographical differences in TTP are needed. PMID- 11726599 TI - Moderate alcohol consumption and waiting time to pregnancy. AB - BACKGROUND: Recent research indicates that even a moderate consumption of alcohol in women trying to become pregnant is associated with longer waiting time to pregnancy. The findings, though, are based upon few observations. METHODS: Self reported data on alcohol intake and waiting time to pregnancy (0-2, 3-5, 6-12 and >12 months) was used for 39 612 pregnant women, recruited to the Danish National Birth Cohort within the first 24 weeks of pregnancy from 1997 to 2000. Main outcome measures were odds ratios (OR) for a prolonged waiting time to pregnancy according to alcohol intake. RESULTS: In nulliparous women neither moderate nor high alcohol intake was related with longer waiting time to pregnancy compared with a low intake. In parous women, a modest association was seen only among those with an intake of >14 drinks per week (subfecundity OR 1.3; 95% confidence interval 1.0-1.7). Women who reported no alcohol intake had a slightly longer waiting time (subfecundity OR 1.2; 95% confidence interval 1.1-1.3) than women with a moderate intake of alcohol. CONCLUSIONS: Our findings do not corroborate recent results suggesting a marked reduction in fecundity associated with a moderate intake of alcohol. PMID- 11726600 TI - Sperm morphological defects related to environment, lifestyle and medical history of 1001 male partners of pregnant women from four European cities. AB - BACKGROUND: Recently, differences in semen quality have been found among the partners of pregnant women from four European cities: Turku, Copenhagen, Edinburgh and Paris. METHODS: During this study, slides from the four centres were subjected to a centralized assessment of sperm morphology. The percentages of sperm defects were recorded using a multiple-entry classification enabling the calculation of the multiple anomalies index (MAI), which is the mean number of anomalies per abnormal sperm. The relationships between various sperm abnormalities and self-reported data on medical history, lifestyle and occupational factors were examined. RESULTS: Significant differences in the MAI and most of the sperm defects were found between the four cities, and more abnormalities were found in spring than in winter. An increase in some sperm abnormalities was related to medical treatment of the mother during pregnancy, higher birthweight and previous treatment for cryptorchidism. Significant variations of several sperm defects were related to stress, weekly working time, occupational posture and metal welding, suggesting directions for further exposure studies. CONCLUSION: The present study indicated that the detailed assessment of sperm abnormalities is a useful biomarker of the effect of various external factors which may qualitatively affect human spermatogenesis. PMID- 11726602 TI - Human blastocyst development media. PMID- 11726601 TI - Evaluation of a sprayable polyethylene glycol adhesion barrier in a porcine efficacy model. AB - BACKGROUND: The formation of adhesions following pelvic surgery remains one of the leading causes of infertility, small bowel obstruction and re-operation for pelvic pain. A novel hydrophilic polyethylene glycol based adhesion barrier (SprayGel) is formed by simultaneously spraying two liquid precursors onto surgical sites. The liquids polymerize to form a gel that effectively coats and adheres to tissue. After about 5 days, the hydrogel layer is absorbed and subsequently undergoes renal clearance. It is believed that the presence of such a barrier would inhibit the formation of adhesions following surgical insult. METHODS: A porcine adhesion model was developed wherein bilateral uterine horn transection and re-anastomosis, along with peritoneal side wall excision was performed via laparotomy. In each animal (n = 10, including the pilot study) one pelvic side wall was treated with adhesion barrier, while the contralateral side remained untreated. RESULTS: At second look laparoscopy, 90% of the untreated sites had adhesions, compared with 30% of the treated sites (P = 0.006). Also observed were statistically significant reductions in the adhesion extent (P = 0.029) and adhesion severity scores (P = 0.023) at the treated sites. However, if the pilot study was excluded (n = 8) the differences obtained were no longer significant. CONCLUSIONS: Polyethylene glycol (SprayGel) merits further investigation as an effective barrier to the formation of post-operative adhesions in this porcine model. PMID- 11726603 TI - Subgroup analyses in Orgalutran trials. PMID- 11726604 TI - Subgroup analyses in Orgalutran trials. PMID- 11726606 TI - Uveal melanoma: no expression of HLA-G. AB - PURPOSE: To investigate whether uveal melanoma cells express HLA-G, a nonclassical HLA class I molecule that has been shown to be a critical mediator in the inhibition of natural killer (NK) cell-mediated cytolysis. METHODS: Eleven human uveal melanoma cell lines were analyzed for the expression of HLA-G by flow cytometry, immunocytochemistry, Western blot analysis, and RT-PCR followed by Southern blot analysis. Two HLA-G-specific monoclonal antibodies were used, 87G and MEM-G/1. In addition, HLA-G expression was determined on frozen tissue sections of 17 primary uveal melanomas. RESULTS: With all HLA-G detection methods, no evidence for HLA-G expression by uveal melanoma cells was found. In contrast, the trophoblast cell line JEG-3 clearly expressed HLA-G transcripts and protein in all cases. Furthermore, interferon-gamma did not induce HLA-G expression in the uveal melanoma cell lines. Notably, all cell lines expressed HLA-E, and this expression was significantly enhanced by interferon-gamma. CONCLUSIONS: Because none of the uveal melanoma cell lines nor any of the primary uveal melanomas displayed expression of HLA-G, it is unlikely that HLA-G plays a role, direct or indirect, in the modulation of cellular immunity against uveal melanoma tumors. PMID- 11726607 TI - Number, distribution, and morphologic particularities of encapsulated proprioceptors in pig extraocular muscles. AB - PURPOSE: To analyze qualitatively and quantitatively the total complement of encapsulated proprioceptors (Golgi tendon organs [GTOs] and neuromuscular spindles) in pig extraocular muscles (EOMs). METHODS: EOMs of four pigs of different ages were prepared for light microscopic histochemical and immunohistochemical analysis and for transmission electron microscopy. RESULTS: GTOs and muscle spindles were numerous in pig EOMs. GTOs were found to be distributed in aponeurotic expansions of the distal and proximal EOM tendons, being more numerous in the distal aponeurosis than in the proximal aponeurosis. The total number of GTOs was higher in the recti EOMs (100-128) than in the oblique EOMs (45-61). Spindles were distributed over the entire muscle length. In each EOM the number of muscle spindles (142-333) exceeded those of GTOs. The morphology of the GTOs was variable. In addition to collagen bundles, approximately one third of the GTOs contained intracapsular muscle fibers that resembled the multiply innervated fiber type. Intracapsular muscle fibers entered the poles of the GTOs and either terminated inside the receptors in collagen bundles or exited the GTOs at the opposite poles. Nerve terminals were numerous in each GTO and established intimate contacts with collagen fibrils. CONCLUSIONS: Most structural particularities formerly observed in GTOs of rhesus monkey and sheep EOMs are also present in GTOs of pig EOMs. The high number of GTOs with their typical nerve terminals indicates functional importance. During muscle activity, afferent signals from GTOs and muscle spindles may provide sufficient information about eye position. PMID- 11726608 TI - Expression of Drosophila omb-related T-box genes in the developing human and mouse neural retina. AB - PURPOSE: To examine the role of Drosophila optomotor blind (omb)-related T-box genes in development of human and mouse retina. METHODS: Mouse Tbx2, Tbx3, and Tbx5 and human TBX2 cDNAs were isolated from retinal cDNA libraries by hybridization to the Drosophila omb gene. Gene expression patterns in developing retina were analyzed by in situ hybridization. RESULTS: TBX2/Tbx2, TBX3/Tbx3, and TBX5/Tbx5 were expressed asymmetrically across the embryonic neural retina with highest levels of mRNA within dorsal and peripheral retina. The dorsoventral gradient of TBX2 expression disappeared before the ganglion cell layer (GCL) formed. Its expression then became restricted to the inner neuroblastic retina and later to the GCL and inner nuclear layer (INL). The dorsal expression domains of TBX5/Tbx5 and TBX3/Tbx3 were maintained during formation of the GCL. As the retina matured, TBX3/Tbx3 expression was restricted to the INL, and TBX5/Tbx5 was expressed within the GCL. CONCLUSIONS: The expression pattern of TBX2, TBX3, and TBX5 within the developing retina supports the idea that the encoded transcription factors play a role in providing positional information important for topographic mapping and in differentiation of distinct cell types across the laminar axis of the retina. PMID- 11726609 TI - Atrophy of Muller glia and photoreceptor cells in chick retina misexpressing cNSCL2. AB - PURPOSE: To investigate whether and how the basic helix-loop-helix (bHLH) gene cNSCL2 is involved in retinal development. METHODS: cNSCL2, the chick homologue of human NSCL2, was isolated and sequenced. In situ hybridization was used to examine its spatial and temporal expression pattern in the retina. Replication competent retrovirus RCAS was used to drive cNSCL2 misexpression in the developing chick retina, and the effect of the misexpression was analyzed. RESULTS: Expression of cNSCL2 in the retina was restricted. Its mRNA was detected in amacrine and horizontal cells, but not in photoreceptor, bipolar, or ganglion cells. Retroviral-driven misexpression of cNSCL2 in the developing chick retina resulted in missing photoreceptor cells and gross deficits in the outer nuclear layer (ONL). These deficits were probably not because of decreased photoreceptor production, in that the ONL appeared normal in early developmental stages. TUNEL+ cells were detected in the ONL, indicating that photoreceptor cells underwent apoptosis in retinas misexpressing cNSCL2. Muller glial cells were far fewer in the experimental retina than in the control, indicating that cNSCL2 also caused Muller glia atrophy. The onset of Muller glia disappearance preceded that of photoreceptor degeneration. CONCLUSIONS: Expression of cNSCL2 in the chick retina was restricted to amacrine and horizontal cells. Misexpression of cNSCL2 caused severe retinal degeneration, and photoreceptor cells and Muller glia were particularly affected. PMID- 11726610 TI - Interaction of the insulin receptor beta-subunit with phosphatidylinositol 3 kinase in bovine ROS. AB - PURPOSE: To identify the tyrosine-phosphorylated protein(s) in bovine rod outer segments (ROS) that are associated with phosphatidylinositol 3-kinase (PI3K). METHODS: Glutathione-S-transferase (GST) fusion proteins containing two SH2 domains of the p85 regulatory subunit of PI3K-GST-p85 (N-SH2), GST-p85 (C-SH2), and respective SH2 mutants (N-SH2, R358A, and C-SH2, R649A)-were prepared and used to pull down tyrosine-phosphorylated proteins in bovine ROS. Protein identity was established by Western blot analysis. PI3K activity was determined in the pull-down mixtures and in immunoprecipitates by incubation with phosphatidylinositol-4,5-bisphosphate (PI-4,5-P(2)) and [gamma(32)P]adenosine triphosphate (ATP). RESULTS: The GST pull-down assays indicated the binding of a 97-kDa protein by GST-p85 (N-SH2) in tyrosine-phosphorylated (PY)-ROS that was not present in nonphosphorylated (N)-ROS. Binding was completely abolished when the Arg 358 in the N-SH2 domain was mutated to Ala. Increased binding of the p110alpha catalytic subunit to GST-p85 (N-SH2) fusion protein was also observed in the presence of the 97-kDa phosphorylated protein. Biochemical evidence indicated that the 97-kDa protein was the beta-subunit of the insulin receptor beta-subunit (IRbeta). Immunoprecipitates of PY-ROS and N-ROS with anti-PY antibodies, probed with anti-IRbeta, indicated the presence of IRbeta only in PY ROS. Immunoprecipitates of PY-ROS and N-ROS with anti-IRbeta antibodies, probed with anti-p85 and anti-p110alpha antibodies, indicated increased amounts of both p85 and p110alpha in PY-ROS compared to N-ROS. Treatment of ROS with insulin, followed by immunoprecipitation with either anti-IRbeta or anti-PY, resulted in increased PI3K activity. Expression and phosphorylation of the cytoplasmic tail of retina insulin receptor showed direct involvement with the p85 subunit of PI3K in vitro. CONCLUSIONS: Tyrosine phosphorylation of the beta-subunit of the insulin receptor is involved in the regulation of PI3K activity in ROS. PMID- 11726611 TI - A novel keratocan mutation causing autosomal recessive cornea plana. AB - PURPOSE: Mutations in keratocan (KERA), a small leucine-rich proteoglycan, have recently been shown to be responsible for cases of autosomal recessive cornea plana (CNA2). A consanguineous pedigree in which cornea plana cosegregated with microphthalmia was investigated by linkage analysis and direct sequencing. METHODS: Linkage was sought to polymorphic microsatellite markers distributed around the CNA2 and microphthalmia loci (arCMIC, adCMIC, NNO1, and CHX10) using PCR and nondenaturing polyacrylamide gel electrophoresis before KERA was directly sequenced for mutations. RESULTS: Positive lod scores were obtained with markers encompassing the CNA2 locus, the maximum two-point lod scores of 2.18 at recombination fraction theta = 0 was obtained with markers D12S95 and D12S327. Mutation screening of KERA revealed a novel single-nucleotide substitution at codon 215, which results in the substitution of lysine for threonine at the start of a highly conserved leucine-rich repeat motif. Structural modeling predicts that the motifs are stacked into an arched beta-sheet array and that the effect of the mutation is to alter the length and position of one of these motifs. CONCLUSIONS: This report describes a novel mutation in KERA that alters a highly conserved motif and is predicted to affect the tertiary structure of the molecule. Normal corneal function is dependent on the regular spacing of collagen fibrils, and the predicted alteration of the tertiary structure of KERA is the probable mechanism of the cornea plana phenotype. PMID- 11726612 TI - Organization of the human IMPG2 gene and its evaluation as a candidate gene in age-related macular degeneration and other retinal degenerative disorders. AB - PURPOSE: To characterize the genomic organization of human IMPG2, the gene encoding the retinal interphotoreceptor matrix (IPM) proteoglycan IPM 200, to evaluate its relationship to IPM 150, and to evaluate its involvement in inherited retinopathies, such as age-related macular degeneration, retinitis pigmentosa, and Leber congenital amaurosis. METHODS: After isolation of human genomic clones, the structure of IMPG2 was determined by sequence analysis. Mutational analyses were conducted on genomic DNA isolated from 316 probands using single-strand conformation polymorphism analysis. RESULTS: The IMPG2 gene is organized into 19 exons, and the structure of the gene is highly similar to that of the IMPG1 gene, which encodes another retinal proteoglycan, IPM 150. Mutational analyses indicate that the observed sequence changes are present at approximately equal rates in donors with and without retinal disease. Additional data derived from RT-PCR and Northern blot analysis show that IMPG2 is processed in the human retina into multiple alternatively sized transcripts that may represent splicing isoforms. CONCLUSIONS: Analysis of the overall relationship of human IMPG2 (located on chromosome 3q12.2-12.3) to human IMPG1 (located on chromosome 6q14) suggests that these genes have evolved from a common ancestral gene. Although this is an excellent candidate gene for hereditary retinopathies, single-strand conformation polymorphism analyses provided no evidence that variations in IMPG2 coding region are responsible for the inherited retinopathies examined. PMID- 11726613 TI - Water-soluble antioxidants in human tears: effect of the collection method. AB - PURPOSE: To resolve differences in published data on tear antioxidant levels by comparing the concentration of water-soluble antioxidants in human reflex tears collected by capillary tube and by the Schirmer strip collection method and in basal and reflex tears collected using the Schirmer strip method. METHODS: Yawn induced reflex tears (collected simultaneously by capillary tubes and by Schirmer strips) and basal tears (by Schirmer strips and using local anesthetic) were collected from 12 healthy subjects. Tear cysteine, ascorbate, glutathione, urate, and tyrosine were measured by high-performance liquid chromatography within a few minutes of collection. RESULTS: Cysteine, ascorbate, glutathione, and tyrosine were 5 to 10 times higher (P < 0.01) in both reflex and basal tears collected by Schirmer strip compared with reflex tears collected by capillary tube from the same subject. Urate levels were slightly but nonsignificantly higher in Schirmer strip samples (P > 0.05). CONCLUSIONS: The conflict in published data on tear antioxidants is caused by differences in collection methods. With the exception of urate, antioxidants accumulate to very high levels in corneal cells. Spuriously high antioxidant levels in tears collected using Schirmer strips, therefore, are most probably caused by contamination with intracellular constituents. The capillary tube collection method is proposed as the method of choice for reflex tear collection for biochemical studies. This less-invasive method facilitates the evaluation of tear antioxidant levels as a biomonitoring tool for corneal health. Although moderately increased antioxidant levels may be beneficial, the authors hypothesize that marked increases may indicate damage to the ocular surface. PMID- 11726614 TI - Maspin: synthesis by human cornea and regulation of in vitro stromal cell adhesion to extracellular matrix. AB - PURPOSE: Maspin, a tumor-suppressor protein that regulates cell migration, invasion, and adhesion, is synthesized by many normal epithelial cells, but downregulated in invasive epithelial tumor cells. The purpose of this study was to determine whether cells in the normal human cornea express maspin and whether maspin affects corneal stromal cell adhesion to extracellular matrix molecules. METHODS: Maspin expression was analyzed by immunodot blot, Western blot, and RT PCR analyses in cells obtained directly from human corneas in situ. Maspin protein and mRNA were also studied in primary and passaged cultures of corneal stromal cells using Western blot analysis, RT-PCR, and immunofluorescence microscopy. Maspin cDNA was cloned and sequenced from human corneal epithelial cells and expressed in a yeast system. The recombinant maspin was used to study attachment of cultured human corneal stromal cells to extracellular matrices. RESULTS: Maspin mRNA and micromolar amounts of the protein were found in all three layers of the human cornea in situ, including the stroma. Maspin was also detected in primary and first-passage corneal stromal cells, but its expression was downregulated in subsequent passages. Late-passage stromal cells, which did not produce maspin, responded to exogenous recombinant maspin as measured by increased cell adhesion not only to fibronectin, similar to mammary gland tumor epithelial cells, but also to type I collagen, type IV collagen, and laminin. CONCLUSIONS: The corneal stromal cell is the first nonepithelial cell type shown to synthesize maspin. Loss of maspin expression in late-passage corneal stromal cells in culture and their biological response to exogenous maspin suggests a role for maspin on the stromal cells in the cornea. Maspin may function within the cornea to regulate cell adhesion to extracellular matrix molecules and perhaps to regulate the migration of activated fibroblasts during corneal stromal wound healing. PMID- 11726615 TI - Role of p27(Kip1) in cAMP- and TGF-beta2-mediated antiproliferation in rabbit corneal endothelial cells. AB - PURPOSE: To determine whether p27(Kip1) plays a role in antiproliferation mediated by antimitogens (cAMP and TGF-beta2) in rabbit corneal endothelial cells (CECs). METHODS: Cell proliferation was assayed using a colorimetric method to determine the number of viable cells. Subcellular localization of cell cycle regulatory proteins was determined by immunofluorescent staining, and expression of the proteins was analyzed by immunoblot analysis. RESULTS: When cells were treated with cAMP or TGF-beta2, serum-mediated cell proliferation was inhibited in a dose-dependent manner. Simultaneous treatment of the two antimitogens did not show a synergistic effect on inhibition of cell growth. Expression of cell cycle-regulatory proteins, such as cyclin-D1, cyclin-E, cdk2, cdk4, p21(Cip1), and p27(Kip1) was determined using immunofluorescent staining. A strong nuclear staining was observed for p27(Kip1). The other proteins were not stained or were only very faintly stained. Treatment of cells with either cAMP or TGF-beta2 did not change the staining potential of any proteins other than p27(Kip1), but all cells were positive for nuclear p27(Kip1) when treated with either TGF-beta2 or cAMP. In contrast, mitogen (FGF-2)-containing medium decreased the number of p27(Kip1)-positive cells. When the expression level of p27(Kip1) was determined using immunoblot analysis in the cells treated either with FGF-2 alone or with a concomitant treatment with FGF-2 and cAMP for 24 hours, FGF-2 markedly decreased the p27(Kip1) level, and cAMP prevented the decrease in p27(Kip1) level induced by FGF-2. No such phenomenon occurred during a short-term exposure of cells to either FGF-2 or cAMP or to a combination of the two. When cells were stained for phosphorylated p27(Kip1), FGF-2 markedly enhanced the staining of phosphorylated p27(Kip1) in nuclei, whereas both cAMP and TGF-beta2 prevented the phosphorylation of p27(Kip1). CONCLUSIONS: These findings suggest that both antimitogens upregulate the expression of p27(Kip1) as they prevent the decrease of the p27(Kip1) level mediated by mitogen. Furthermore, cAMP and TGF-beta2 may inhibit the G(1)-to-S transition by blocking phosphorylation of p27(Kip1), which is a prerequisite for nuclear export of the inhibitor molecule for degradation. PMID- 11726616 TI - Predicting extended wear complications from overnight corneal swelling. AB - PURPOSE: To examine the hypothesis that the corneal overnight swelling response (ONSR) is a predictor of ocular complications in contact lens extended wear (EW). METHODS: The Berkeley Contact Lens Extended Wear Study (CLEWS) was a randomized, concurrently controlled clinical trial in which more than 200 subjects in EW with rigid gas-permeable (RGP) lenses were observed for 1 year. After adapting to EW, subjects were randomized to either medium or high oxygen-permeable (Dk) RGP lenses and underwent clinical assessments, keratometry, and corneal pachometry at 3-month intervals. RESULTS: The ONSR was directly related to lens Dk (P = 0.01) and exhibited substantial variability across subjects. The probability of remaining free of complications over time was not significantly lower for subjects with a mild ONSR compared with those with greater edema (P = 0.84). The risk of development of keratopathy was not significantly related to the ONSR (relative risk = 1.00). CONCLUSIONS: The corneal ONSR is not a good predictor of ocular complications in 1 year of RGP EW. Lenses that cause little or no corneal edema are not necessarily safer for overnight wear. PMID- 11726617 TI - Long-term changes in myosin heavy chain composition after botulinum toxin a injection into rat medial rectus muscle. AB - PURPOSE: To study long-term changes of extraocular muscles after botulinum toxin (Botx) A-induced paralysis, with special emphasis on myosin heavy chain (MyHC) isoform pattern in muscle fibers. METHODS: Botx A (5 IU) was injected into the ocular medial rectus (MR) muscles of adult rats. After 1, 5, and 8 months muscle cross sections were examined immunohistochemically, histochemically, and morphometrically. MyHC content was analyzed by gel electrophoresis. RESULTS: Paralyzed MR muscles displayed mildly atrophic and hypertrophic muscle fibers and decreased oxidative metabolism, due to decreased succinate dehydrogenase activity. However, muscle morphology was not grossly disturbed. MyHC profile was shifted toward slower isoforms. Electrophoretic analysis showed that the share of MyHCI, and especially of MyHCIIa and MyHCIIx/d, increased several fold, whereas the share of MyHCIIb decreased heavily during the first 5 months. Immunohistochemical analysis generally mirrored the results obtained by electrophoresis. Moreover, specific extraocular MyHC isoform MyHCeom disappeared and could not be detected during the whole experimental period. The portion of MyHCIIb relatively increased 8 months after Botx A injection, although the MyHC profile was still far from normal. CONCLUSIONS: These long-lasting changes in Botx A-paralyzed ocular MR muscles most probably reflect their inability to regain their unique functional characteristics after new motor end plate formation and recovery of muscle contraction. PMID- 11726618 TI - Changes in mRNA levels of the Myoc/Tigr gene in the rat eye after experimental elevation of intraocular pressure or optic nerve transection. AB - PURPOSE: To isolate the rat Myoc/Tigr gene and investigate changes in its expression pattern in normal eyes and in eyes with either pressure-induced optic nerve damage or optic nerve transection. METHODS: Expression pattern of the rat Myoc/Tigr gene was investigated by Northern blot hybridization. Optic nerve damage and death of ganglion cells in the retina were induced unilaterally, by injection of hypertonic saline solution, episcleral vein cauterization, or optic nerve transection. The levels of mRNA for Myoc/Tigr were compared between several tissues of the control and surgically altered eyes, by using semiquantitative RT PCR, real-time PCR, and Northern blot analysis. RESULTS: The rat Myoc/Tigr gene is 10 kb long and contains three exons. Among the eye tissues analyzed, Myoc/Tigr mRNA was detected in the combined tissues of the eye angle, sclera, cornea, retina, and optic nerve head. With pressure-induced optic nerve degeneration, the level of Myoc/Tigr mRNA decreased in the retina and the combined tissues of the eye angle, but increased in the optic nerve head. After optic nerve transection, the level of Myoc/Tigr mRNA increased in the retina, but did not change in the combined tissues of the eye angle. CONCLUSIONS: The decreased level of Myoc/Tigr mRNA in the retina after induction of elevated intraocular pressure compared with that in the control retina cannot be explained by ganglion cell death alone. Differences in Myoc/Tigr mRNA levels in eye tissues after elevation of intraocular pressure or optic nerve transection may reflect the activation of different signaling pathways involved in regulation of this gene. PMID- 11726619 TI - Delayed secondary glucocorticoid responsiveness of MYOC in human trabecular meshwork cells. AB - PURPOSE: To characterize the glucocorticoid responsiveness of the glaucoma gene MYOC (myocilin/TIGR) in cultured human trabecular meshwork (TM) cells. METHODS: MYOC expression in two independently derived human TM cell lines was quantified by Western immunoblot analysis of protein levels and quantitative PCR analysis of mRNA levels. Promoter activity was measured indirectly with the luciferase reporter gene in a dual luciferase reporter assay. RESULTS: Application of the synthetic glucocorticoid dexamethasone (Dex) to cultured TM cells at 100 nM resulted in a delayed (8-16 hours) induction of myocilin. The concentration dependence (median effective concentration [EC(50)], approximately 10 nM) and reversal by the glucocorticoid antagonist, RU486, implicates the glucocorticoid receptor (GR). In an interesting observation, RU486 alone acted as a partial agonist to MYOC expression. Treatment of TM cells with the protein synthesis inhibitor cycloheximide abolished the Dex induction, suggesting an indirect effect of the GR on MYOC expression. In addition, the RNA synthesis inhibitor actinomycin D also blocked Dex induction, indicating that the Dex effect was due to increased MYOC transcription. Analysis of up to 2700 nucleotides (nt) of the MYOC gene 5'-flanking region in luciferase reporter constructs showed no Dex induction, despite the presence of multiple putative glucocorticoid response element (GRE)-like half-sites in the MYOC promoter and the presence of an intact cellular GR-mediated signaling system. CONCLUSIONS: MYOC is a delayed secondary glucocorticoid-responsive gene. Characterization of the transcription factors that mediate the secondary response will shed new light on the pathophysiology of steroid-induced ocular hypertension and glaucoma. PMID- 11726620 TI - Detection of prostaglandin EP(1), EP(2), and FP receptor subtypes in human sclera. AB - PURPOSE: To examine the expression of five prostaglandin (PG) receptors, EP(1), EP(2), EP(3), EP(4), and FP and their corresponding mRNA transcripts in human sclera and cultured human scleral fibroblasts (HSFs). METHODS: Primary cultures of HSFs were established from donor eyes. Also, sclera from human donor eyes was snap frozen and sectioned. Immunocytochemistry was performed on HSFs and tissue sections with subtype-specific antibodies to the EP(1), EP(2), EP(3), EP(4), and FP receptors. The presence of mRNA for the receptor subtypes was examined from total RNA obtained from human sclera and confirmed with restriction digest analysis. RESULTS: Positive EP(1) and FP receptor immunoreactivity was observed in fibroblasts within the sections from human sclera. In primary cultures of HSFs, EP(1) and FP labeling was observed over the entire cell surface. EP(2) immunoreactivity within HSFs was mostly present in the juxtanuclear region. RT PCR analysis of total RNA isolated from human sclera and HSFs confirmed the presence of EP(1), EP(2), and FP receptor subtypes. The identity of the polymerase chain reaction products was confirmed by restriction enzyme analysis. No mRNA or immunoreactivity above basal levels was detected for the EP(3) and EP(4) prostanoid receptor subtypes in tissue sections or primary cultures. CONCLUSIONS: The EP(1), EP(2), and FP receptor subtypes are present in HSFs, suggesting that these cells may respond to endogenous PGs and their structural analogues through interaction with these receptor subtypes. PMID- 11726621 TI - Effects of the marine macrolides swinholide A and jasplakinolide on outflow facility in monkeys. AB - PURPOSE: To determine effects of the marine macrolides swinholide A (Swin A) and jasplakinolide (Jas), alone or in conjunction with latrunculin B (Lat B) on outflow facility in monkeys. METHODS: Total outflow facility was measured by two level constant-pressure perfusion of the anterior chamber before and after exchange with Swin A, Jas, or vehicles followed by continuous anterior chamber infusion of drug or vehicle, in opposite eyes of cynomolgus monkeys. The effect of a facility-ineffective dose of Jas plus a threshold or submaximal facility effective dose of the actin depolymerizer Lat B on outflow facility was also determined. RESULTS: Ten or 100 nM Swin A or 20, 100, or 500 nM Jas had no significant effect on outflow facility. However, 500 nM Swin A and 2.5 microM Jas significantly increased facility by 80% +/- 21% and 157% +/- 57% (mean +/- SEM) respectively, adjusted for corresponding baselines and resistance washout in contralateral control eyes. The facility increase in the eye treated with 500 nM Jas with 60 or 200 nM Lat B was similar to that in the eye treated with 60 or 200 nM Lat B only. CONCLUSIONS: Swin A (which severs actin filaments and sequesters actin dimers) and Lat B (which sequesters actin monomers) similarly increase outflow facility. The potent inducer of actin polymerization Jas (500 nM) neither inhibits nor potentiates the facility increase induced by Lat B (60 or 200 nM). A higher dose of Jas increases rather than decreases outflow facility. PMID- 11726622 TI - Mechanisms of action of unoprostone on trabecular meshwork contractility. AB - PURPOSE: This study was performed to clarify the possible mechanism behind the ocular hypotensive effect of unoprostone isopropyl (Rescula; Novartis Ophthalmics AG, Basel, Switzerland), a new docosanoid that has been shown to reduce intraocular pressure (IOP) in patients with ocular hypertension or primary open angle glaucoma. To gain insight into the possible mode of action, the effects of unoprostone on ciliary muscle (CM) and trabecular meshwork (TM) contractility, intracellular calcium levels, and membrane channels were investigated. METHODS: The effects of unoprostone (M1 metabolite = free acid, 10(-5) M) and endothelin (ET)-1 (10(-9) M) on bovine TM (BTM) and ciliary muscle (CM) strips were investigated, by using a custom-made force-length transducer system. The effects of unoprostone and ET-1 (5 x 10(-8) M) on intracellular Ca(2+) mobilization in cultured human TM (HTM) were measured using fura-2AM as a fluorescent probe. Patch-clamp experiments were performed on HTM and BTM cells to investigate the unoprostone-dependent modulation of membrane currents. RESULTS: In isolated TM and CM strips, unoprostone almost completely inhibited ET-induced contractions (TM: 2.9% +/- 4.3% vs. 19.6% +/- 5.7%, P < 0.05, n = 6; CM: 1.4% +/- 1.6% vs. 30.1% +/- 5.3%, P < 0.01, n = 6; 100% = maximal carbachol-induced (10(-6) M) contraction). However, neither carbachol-induced contraction nor baseline tension was affected by unoprostone. Furthermore, unoprostone had no effect on baseline intracellular calcium levels (baseline: 126 +/- 45 nM versus unoprostone: 132 +/- 42 nM, n = 8) in HTM cells. The endothelin-induced increase (679 +/- 102 nM), however, was almost completely (P < 0.01) blocked by unoprostone (178 +/- 40 nM). In patch-clamp recordings, unoprostone could be shown to double the amplitude of outward current (HTM: 200% +/- 33%; n = 6; BTM: 179% +/- 20%; n = 8). This effect was blocked by the specific inhibitor of maxi-K channels, iberiotoxin. CONCLUSIONS: This study presents evidence for direct interaction of unoprostone with the contractility of the TM and CM. This compound may lower IOP by affecting aqueous outflow, most probably conventional outflow pathways (i.e., TM) through inhibition of ET-dependent mechanisms. In addition, unoprostone interacts with the maxi-K channel. Although primarily Ca(2+)-sensitive signal-transduction pathways seem to be involved, effects of unoprostone on Ca(2+)-independent pathways and uveoscleral outflow cannot be excluded. PMID- 11726623 TI - Posterior scleral thickness in perfusion-fixed normal and early-glaucoma monkey eyes. AB - PURPOSE: To characterize posterior scleral thickness in the normal monkey eye and to assess the effects of acute (15- to 80-minute) and short-term chronic (3- to 7 week) intraocular pressure (IOP) elevations. METHODS: Both eyes of four normal monkeys (both eyes normal) and four monkeys with early glaucoma (one eye normal and one eye with induced chronic elevation of IOP) were cannulated. In each monkey, IOP was set to 10 mm Hg in the normal eye and 30 or 45 mm Hg in the contralateral eye (normal or early glaucoma) for 15 to 80 minutes. All eight monkeys were perfusion fixed, yielding eight low IOP-normal eyes, four high IOP normal eyes, and four high IOP-early glaucoma eyes. Posterior scleral thickness was measured histomorphometrically at 15 measurement points within each eye, and the data were grouped by region: foveal, midposterior, posterior-equatorial, and equatorial. RESULTS: Overall, posterior scleral thickness was significantly different in the various regions and among the treatment groups (P < 0.0001). In the low IOP-normal eyes, the posterior sclera was thickest in the foveal region (307 microm) and thinner in the midposterior (199 microm), posterior-equatorial (133 microm), and equatorial (179 microm) regions. In the high IOP-normal and high IOP-early glaucoma eyes, the posterior sclera was thinner both overall and within specific regions, compared with the low IOP-normal eyes. CONCLUSIONS: The posterior sclera in the perfusion-fixed normal monkey eye thins progressively from the fovea to the equator and is thinnest just posterior to the equator. Acute and short-term chronic IOP elevations cause regional thinning within the posterior sclera of some monkey eyes, which significantly increases stresses in the scleral wall. PMID- 11726624 TI - Prostanoid receptor gene expression profile in human trabecular meshwork: a quantitative real-time PCR approach. AB - PURPOSE: To assess the expression pattern of prostanoid receptor-encoding genes in trabecular meshwork (TM) of human donor eyes. METHODS: Disposed human donor eyes (n = 10) were obtained from the Cornea Bank, Amsterdam. The TM was dissected from the scleral tissue and homogenized in lysis buffer, and total RNA was isolated. The RNA was converted into cDNA and used as a template for noncompetitive quantitative real-time polymerase chain reaction (PCR) using green fluorescent dye to quantify the accumulation of double-stranded PCR product. Specific primers for four housekeeping genes and DP, EP(1), EP(2), EP(3,) EP(4), FP, IP, and TP receptor-encoding transcripts were developed and tested for their efficiency. RESULTS: The characterized expression profile was highly reproducible in all samples, with the EP(2) receptor-encoding transcript in the highest abundance, followed by FP, TP, IP, and EP(4) at levels that were approximately 10 to 15 times lower than that of the EP(2) subtype. DP and EP(3) were at the lowest levels, which were, on average, 45 times and 228 times lower than EP(2), respectively. CONCLUSIONS: These data show that all prostanoid receptors are expressed at different levels in human TM tissue. Because the gene expression of the EP(2) receptor is, on average, 15 times more abundant than that of the EP(4) receptor, it may be expected that the increase in flow and cAMP levels in response to the activation of the EP receptors by application of prostaglandin E(1) (PGE(1)), is primarily mediated by the EP(2) receptor. These data should be considered when designing prostanoid receptor mimetics intended to enhance the aqueous humor outflow through the TM and Schlemm's canal. PMID- 11726625 TI - Atrophy of relay neurons in magno- and parvocellular layers in the lateral geniculate nucleus in experimental glaucoma. AB - PURPOSE: To determine whether in glaucoma there is atrophy of relay neurons in magnocellular and/or parvocellular lateral geniculate nucleus (LGN) layers projecting to the visual cortex and to compare the degree of neuronal atrophy in magnocellular layers with that in parvocellular layers. METHODS: Seven cynomolgus monkeys with unilateral experimentally induced glaucoma and five control monkeys were studied. The left LGN neurons in magnocellular layer 1 and parvocellular layers 4 and 6, connected to the right glaucomatous eye were examined. Immunocytochemistry with antibody to parvalbumin was used to specifically label relay neurons connecting to the visual cortex. Neuronal cell body cross-sectional area was estimated using unbiased point-counting methodology. Experimental and control groups were compared using t-tests. Analysis of covariance (ANCOVA) tests were used to compare the percentage of decrease in mean neuronal area between layers 1, 4, and 6, as a function of percentage of optic nerve fiber loss or mean IOP. There was significant correlation between percentage of optic nerve fiber loss and mean IOP. RESULTS: The mean cross-sectional area of relay neurons in magnocellular layer 1 and parvocellular layers 4 and 6 were significantly decreased in glaucoma compared with controls by 28%, 37%, and 45%, respectively. Neuronal area decreased in a linear fashion, with increasing optic nerve fiber loss or increasing mean IOP for layers 1, 4, and 6. The percentage of neuronal shrinkage in each of parvocellular layers 4 and 6, as a function of optic nerve fiber loss (P = 0.05; P = 0.001, respectively) or mean IOP (P = 0.046; P = 0.0008, respectively), was greater than that seen in magnocellular layer 1. CONCLUSIONS: Relay neurons in the LGN, which project to the visual cortex, undergo significant shrinkage in glaucoma, and neurons in parvocellular layers undergo significantly more shrinkage than neurons in magnocellular layers. PMID- 11726626 TI - Expression of membrane-type matrix metalloproteinases 4, 5, and 6 in mouse corneas infected with P. aeruginosa. AB - PURPOSE: To investigate the expression and regulation of membrane-type matrix metalloproteinases (MT-MMPs) 4, 5, and 6 in the mouse corneas infected with Pseudomonas aeruginosa. METHODS: C57BL/6J mice were intracorneally infected with P. aeruginosa. The expression of MT4-, MT5-, and MT6-MMP was detected at both the mRNA and protein levels by RT-PCR and immunoblot analysis. Immunohistochemical staining was performed to localize the expression of MT4- and MT5-MMP in the mouse corneas. RESULTS: Expression of MT4- and MT5-MMP was detected in the normal (uninfected) cornea by RT-PCR and immunoblot analysis. When infected with P. aeruginosa, the corneas showed significant induction of each MT-MMP. Localization of MT4- and MT5-MMP revealed that the expression of MT5-MMP was restricted to the epithelial tissue in the normal cornea, whereas the induced expression of MT4- and MT5-MMP was predominantly in the substantia propria, which contained most of the infiltrating cells. MT6-MMP expression was not detected in the uninfected cornea but was upregulated in the infected corneas. CONCLUSIONS: Expression of MT4-, MT5-, and MT6-MMP was induced in corneas infected with P. aeruginosa. Immunohistochemistry showed predominant immunoreactivity of MT4- and MT5-MMP in the substantia propria. Previous histologic studies have revealed different patterns of inflammatory cell infiltration with an increased number of polymorphonuclear neutrophils (PMNs) during the early stage of inflammation and increased macrophages during the late stage. These results indicate a good correlation between the overexpression of the MT-MMPs in the infected corneas and the inflammatory response-that is, leukocyte infiltration-indicating that inflammatory cells such as macrophages and PMNs may play a role in the upregulation of MT-MMPs during corneal infection, which in turn can cause the destruction of corneal tissue. PMID- 11726627 TI - Characterization of Toxoplasma gondii-specific T cells recovered from vitreous fluid of patients with ocular toxoplasmosis. AB - PURPOSE: The mechanisms involved in reactivations of latent ocular Toxoplasma gondii (Tg) infections in immunocompetent patients are poorly understood. In view of the possible role of T cells in the immunopathogenesis of the disease, ocular infiltrating T cells obtained from patients with recurrent ocular toxoplasmosis were characterized phenotypically and functionally. METHODS: Ocular infiltrating T cells were recovered from vitreous fluid (VF) samples of 10 patients with active recurrent ocular toxoplasmosis. Two patients with uveitis of other origins were included as control subjects. T-cell lines (TCLs) were generated by mitogenic stimulation and tested for reactivity to Tg and human retinal protein extracts. The TCLs of three patients were cloned by limiting dilution. Tg reactive T-cell clones (TCCs) were characterized with respect to their phenotype, T-cell receptor variable (TCR V)-beta gene usage, HLA restriction, and cytokine secretion profile. RESULTS: Reactivity to Tg could be detected only in the TCLs of patients with ocular toxoplasmosis. None of the TCLs showed reactivity to human retinal antigens. All tested intraocular Tg-specific TCCs (n = 23) were CD3+CD4+ and displayed differential TCR Vbeta usage. Twenty-one TCCs were HLA-DR restricted and two TCCs were restricted by HLA-DP. The majority of the intraocular Tg-specific TCCs showed a bias toward a T-helper (Th)0-Th2 cytokine profile. CONCLUSIONS: The data indicate that T cells specific for the triggering microorganism infiltrate the eye of patients with recurrent ocular toxoplasmosis. The functional characteristics of the VF-derived Tg-specific T cells and their presence at the site of inflammation suggest their involvement in the local inflammatory response of ocular toxoplasmosis. PMID- 11726628 TI - Human S-antigen determinant recognition in uveitis. AB - PURPOSE: Soluble antigen (S-Ag) is a member of the arrestin family of protein with which it shares a high level of homology. It is an immunologically privileged retinal antigen that can elicit experimental autoimmune uveitis (EAU) and is thought to be a target for ocular inflammatory diseases. This study was conducted to identify in humans, the immunogenic determinants of human S-Ag and to establish whether a specific response profile occurs in particular ocular inflammatory conditions. METHODS: Peripheral blood lymphocyte responses were measured against a panel of 40 overlapping synthetic peptides of human S-Ag in patients with chronic uveitis and compared with control subjects. Patients with Behcet disease, sarcoidosis, Vogt-Koyanagi-Harada, and sympathetic ophthalmia were tested. RESULTS: A limited number of immunodominant determinants were identified for Behcet disease and sarcoidosis. These were all located at sites of limited homology with other known arrestins. In addition, several individual patients had prominent proliferative responses to multiple determinants well above that of control subjects. This determinant spread was observed in all disease entities except sympathetic ophthalmia, which did not show any immunoreactivity to S-Ag. Significant response shifts were also noted over time in two patients. CONCLUSIONS: The results indicate that there are specific immunodominant determinants to human S-Ag in patients with certain forms of uveitis. However, in individual patients, response is not limited to these determinants. In the chronic stage of disease, response is spread over many determinants. PMID- 11726629 TI - Regulation of c-fos induction in lens epithelial cells by 12(S)HETE-dependent activation of PKC. AB - PURPOSE: 12(S)-Hydroxyeicosatetraenoic acid (12(S)HETE), a 12-lipoxygenase metabolite of arachidonic acid, is required for epidermal growth factor (EGF) dependent DNA synthesis and c-fos induction in lens epithelial cells. The present study was undertaken to identify signal transduction events upstream of c-fos induction that may be regulated by 12(S)HETE. METHODS: The rabbit lens epithelial cell line, N/N1003A, was cultured in serum-free medium, with or without EGF. Activation of PKC and other selected enzymes was examined in the presence of the lipoxygenase inhibitor baicalein and/or exogenous 12(S)HETE. Relative abundance of PKC isoforms in subcellular fractions was determined by immunoblot analysis with isoform-specific antibodies. PKC activity in subcellular fractions was measured by peptide substrate phosphorylation, with and without pseudosubstrate peptide inhibitor. Phosphorylated enzymes were detected by immunoblot analysis. Relative levels of c-fos mRNA were determined by RT/PCR with internal standard. RESULTS: Baicalein blocked EGF-dependent translocation and activation of PKC, without affecting phosphorylation of Erk1/2. Of several PKC isoforms investigated (alpha, betaI, betaII, and gamma), only PKCalpha and betaII were significantly activated by EGF and inhibited by baicalein. 12(S)HETE, in combination with EGF, countered the effect of lipoxygenase inhibitors on PKC activation, and 12(S)HETE in the absence of EGF stimulated PKC translocation. Also of note, 12(S)HETE alone activated PKCgamma, an isoform that was not significantly activated by EGF. Inhibiting PKC activation with GF109203X blocked induction of c-fos by EGF but did not affect EGF-stimulated phosphorylation of Erk1/2, indicating that the effect of PKC on c-fos induction is independent of the Erk1/2 pathway. CONCLUSIONS: In lens epithelial cells, 12(S)HETE-dependent activation of PKCalpha and betaII acts in concert with other EGF-dependent signals to induce c-fos mRNA. PMID- 11726630 TI - Glutathione peroxidase-1 deficiency leads to increased nuclear light scattering, membrane damage, and cataract formation in gene-knockout mice. AB - PURPOSE: Previous in vitro studies with transgenic and gene-knockout mice have shown that lenses with elevated levels of glutathione peroxidase (GPX)-1 activity are able to resist the cytotoxic effect of H(2)O(2), compared with normal lenses and lenses from GPX-1-deficient animals. The purpose of this study was to investigate the functional role of this enzyme in antioxidant mechanisms of lens in vivo by comparing lens changes of gene-knockout mice with age-matched control animals. METHODS: In vivo lens changes were monitored by slit lamp biomicroscopy, and enucleated lenses were examined under a stereomicroscope in gene-knockout animals and age-matched control animals ranging in age from 3 weeks to 18 months. Transmission (TEM) and confocal microscopy were performed on different regions of lenses after the mice were killed at various times. RESULTS: Slit lamp images showed an increase in nuclear light scattering (NLS) in gene-knockout mice compared with control animals. TEM revealed changes in the nucleus as early as 3 weeks of age by the appearance of waviness of fiber membranes. With increasing age, there was greater distortion of fiber membranes and distension of interfiber space at the apex of fiber cells compared with control mice. The changes in nuclear fiber membranes were even more dramatic, as observed by confocal microscopy, which was performed on thicker sections. In contrast to the changes in the lens nucleus, the morphology of the epithelium and superficial cortex remained unchanged in knockout animals during the same experimental period, consistent with slit lamp observations. Stereomicroscopy of ex vivo lenses demonstrated a significant increase in opacification in gene-knockout mice relative to control animals of the same age. This effect became evident in mice aged 5 to 9.9 months and persisted thereafter in older animals, resulting in mature cataracts after 15 months. CONCLUSIONS: The results demonstrate the critical role of GPX-1 in antioxidant defense mechanisms of the lens nucleus. The increased NLS appears to be associated with damage to fiber membranes in the nucleus, which is particularly susceptible to oxidative challenge because of the deficiency of GPX-1. It is suggested that the lens membrane changes in the knockout animals may be due to the formation of lipid peroxides, which serve as substrates for GPX-1. Cataract development in gene-knockout mice appeared to progress from focal opacities, apparent at an earlier age, to lamellar cataracts between 6 and 10 months, and finally to complete opacification in animals older than 15 months. This is the first reported phenotype in GPX-1-knockout mice. PMID- 11726631 TI - Immunocytochemical characterization of cysts in the peripheral retina and pars plana of the adult primate. AB - PURPOSE: To better characterize the cellular constituents of cysts in the peripheral retina and pars plana of the adult monkey. METHODS: Frozen sections of the peripheral retinal margin and pars plana from monkeys (Macaca nemestrina) between 1 and 15 years of age were stained with toluidine blue or immunolabeled with a variety of glia- and neuron-specific antibodies. RESULTS: In animals 1 to 2 years of age, the nonpigmented inner layer of the pars plana is a pseudostratified columnar epithelium. In these young animals, the peripheral retina had distinct layers and did not contain cysts. In animals 6 years of age or older, there were numerous cysts in the pars plana and in the peripheral retina. In the peripheral retina, neurons were randomly distributed and did not have a laminar organization. Cells surrounding cysts were immunoreactive for different types of markers for retinal neurons. Some of the cells surrounding cysts in the pars plana were also unexpectedly immunoreactive for antigens normally expressed only in retinal neurons and glia. CONCLUSIONS: Cysts form in the peripheral retina and pars plana in adult monkeys. The peripheral retinal cysts disrupt the normal lamination of the cells, but all types of retinal neurons are still present in the cysts. In an unexpected finding, cysts in the pars plana also contained cells immunoreactive for a few of the markers of retinal cells, suggesting that neurogenesis may occur in the pars plana of the adult primate. PMID- 11726632 TI - The ability of hyperoxia to limit the effects of experimental detachment in cone dominated retina. AB - PURPOSE: To determine the ability of oxygen supplementation to ameliorate the effects of retinal detachment in a cone-dominated retina. METHODS: Retinal detachments were created in the right eyes of ground squirrels and the animals immediately placed in normoxic (room air) or hyperoxic (70% oxygen) conditions for 3 days. The retinas were sampled from different regions and investigated morphologically or immunocytochemically by light or confocal microscopy. Agarose embedded sections were immunostained with antibody probes to cytochrome oxidase, synaptophysin, medium-to-long wavelength-sensitive (M/L) cone opsin, rod opsin, excitatory amino acid transporter 1 (EAAT1), glutamate synthetase (GS), cellular retinaldehyde-binding protein (CRALBP), and peanut agglutinin (PNA) lectin. Retinal wholemounts were labeled with PNA and antibodies to short (S)-wavelength sensitive cone opsin and rod opsin. Cell death was examined using a TUNEL assay on agarose sections or using toluidine blue staining on semithin sections. RESULTS: The percentage of dying cells relative to the total nuclei in the photoreceptor layer was significantly reduced, and the total number of nuclei was greater in hyperoxic animals. Triple labeling using TUNEL, anti-M/L cone opsin and anti-rod opsin showed that hyperoxia had a remarkable effect both on the reduction of cone cell death and the maintenance of the overall structure of cone photoreceptors. Analysis of the retinal wholemounts demonstrated the preservation of PNA, S-cone, and rod opsin antibody labeling in the detachments maintained in hyperoxic conditions. Although the disruption of cytochrome oxidase and synaptophysin was seen in normoxic animals, there was minimal disruption in hyperoxic animals. Labeling with anti-EAAT1, anti-GS, and anti-CRALBP was increased in the Muller cells of normoxic animals with detachments, but was decreased in the hyperoxic animals. CONCLUSIONS: Hyperoxia prevents the degeneration of both rods and cones in retinas heavily dominated by cones and mitigates the effect of detachment on Muller cell reactivity. The current results suggest that the rescue of cones is not secondary to that of rods. PMID- 11726633 TI - Polarity and developmental regulation of two PDZ proteins in the retinal pigment epithelium. AB - PURPOSE: Identification of binding partners for ezrin, an actin-binding protein crucial for morphogenesis of apical microvilli and basolateral infoldings in RPE cells. METHODS: Rat eyes, rat primary RPE, the rat RPE-J cell line, and a clonal line of RPE-J cells transfected with human ezrin cDNA were analyzed by immunofluorescence microscopy and immunoblot. Immunofluorescence localization of two ezrin-binding proteins was performed in cryosections of rat eyes of various ages and in monolayers extracted with the detergent Triton X-100 and fixed in paraformaldehyde. The interaction of both proteins with ezrin and gluthathione-S transferase (GST)-ezrin fusion proteins was analyzed by SDS-PAGE and immunoblot. RESULTS: Immunofluorescence microscopy of adult rat eyes detected a polarized distribution of ERM (ezrin, radixin, and moesin)-binding phosphoprotein of 50 kDa (EBP50) at the apical microvilli and synaptic-associated protein of 97 kDa (SAP97) at the basolateral surface of RPE cells, which overlapped with ezrin. These two PDZ (postsynaptic density protein [PSD-95]/disc large [DLG]-A/ZO-1) domain proteins had a similar polarized distribution and high resistance to detergent extractability, indicative of cytoskeletal association, both in primary cultures of rat RPE and in a clonal RPE-J cell line expressing high levels of transfected ezrin. RPE cell lysates from rat retinas of various postnatal ages revealed increasing levels of EBP50 and SAP97 compared with alphav integrin, a protein expressed at constant adult levels from birth. GST pull-down and immunoprecipitation experiments demonstrated a direct interaction between EBP50 and SAP97 and ezrin. CONCLUSIONS: The data indicate that EBP50 localizes at the apical microvilli, whereas SAP97 localizes at the basolateral surface of RPE cells, probably through a direct interaction with ezrin. PMID- 11726634 TI - Protection of retinal ganglion cells from ischemia-reperfusion injury by electrically applied Hsp27. AB - PURPOSE: To determine whether the Hsp27 protein can rescue retinal ganglion cells (RGCs) of rats from ischemia-reperfusion injury. METHODS: Retinal ischemia was induced in rats by clamping the ophthalmic artery within the dural sheath of the optic nerve. Immediately after removing the clamp and beginning the reperfusion, Hsp27 protein solution was injected into the vitreous, and electroporation was applied. To determine whether Hsp27 entered the RGCs, anti-Hsp27 immunohistochemistry was performed. The retinal damage was evaluated by counting the number of RGCs retrogradely labeled by 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine percholorate (diI) injected into the superior colliculus, and also by comparing the ratio of TUNEL-positive to all RGCs in the RGC layer. RESULTS: Electroporation successfully delivered Hsp27 protein into RGCs. In the Hsp27 electroinjected group, the number of RGCs 7 days after ischemia-reperfusion was significantly higher than in the control groups. The ratio of TUNEL-positive cells to all RGCs was lower in the group electroinjected with Hsp27 protein. CONCLUSIONS: Electroporation of Hsp27 protein into RGCs increased the resistance of the RGCs to the apoptosis induced by ischemia reperfusion injury. PMID- 11726635 TI - Evidence for pigment epithelium-derived factor receptors in the neural retina. AB - PURPOSE: The neurotrophic activity of pigment epithelium-derived factor (PEDF), an extracellular factor present in the retina, is mediated by binding to cell surface receptors in responsive cell cultures. In the present study, the expression of PEDF receptors in native neural retinas from adult steers was examined. METHODS: Binding reactions were performed with (125)I-PEDF and fluoresceinated PEDF using plasma membranes, detergent-soluble membrane proteins, or cryosections of retina from adult bovine eyes. Radioligand-binding and competition analyses were performed with a computer-assisted program. Ligand blot analysis of detergent-soluble membrane proteins was performed with (125)I-PEDF followed by autoradiography. Ligand-affinity column chromatography of detergent soluble membrane proteins was performed with PEDF-coupled resin followed by SDS PAGE. Binding of fluoresceinated PEDF to retina cryosections was detected by confocal microscopy. RESULTS: Radioligand-binding assays showed that (125)I-PEDF bound in a specific and saturable fashion to one class of sites on retina membranes (K(d) = 2.5-6.5 nM; maximum binding [B(max)] = 1-48 x 10(10) sites/retina). A peptide of 44 amino acids (44-mer), identified as the receptor binding region of PEDF, competed efficiently for (125)I-PEDF binding to retina membranes with kinetics similar to the full-length PEDF. Ligand blot analysis and ligand-affinity chromatography revealed a specific and high-affinity PEDF-binding protein of approximately 85 kDa in retina plasma membranes. Confocal microscopy showed that fluorescein-conjugated PEDF stained exclusively the inner segments of photoreceptors and cells of the ganglion cell layer in retinal cryosections. CONCLUSIONS: Altogether, these data conclusively demonstrate the existence of PEDF receptors discretely distributed on the surface of cells from the adult neural retina of bovine eyes. Furthermore, they provide evidence for the direct action of PEDF on photoreceptor and ganglion cell neurons and an anatomic basis for studies to assess PEDF neurotrophic effects on the adult retina. PMID- 11726636 TI - Gene transfer mediated by recombinant baculovirus into mouse eye. AB - PURPOSE: To determine the efficiency of baculoviruses (BVs) to transfer recombinant genes in vivo into murine ocular tissues. METHODS: Recombinant (r)BVs carrying fluorescent protein (FP) cDNA under the control of cytomegalovirus (CMV) immediate early promoter were constructed. Initially, cultured HEK293 and ARPE19 cells were infected with these rBVs and analyzed for efficiency and stability of transgene expression. The rBV-CMV green (G)FP was also injected into the intravitreal and subretinal space of mouse eye. Mice were periodically analyzed to determine the efficiency and stability of expression by histologic examination under fluorescence microscopy. The effect of rBV-CMV-GFP on the physiology of the retina was analyzed by electroretinography. RESULTS: cDNAs encoding fluorescent proteins were efficiently transduced in HEK293 and ARPE19 cells in vitro. GFP expression in vivo was observed exclusively in retinal pigment epithelial (RPE) cells after subretinal injections. Intravitreal injections of rBV resulted in GFP expression in the corneal endothelium, lens, RPE, and retina. GFP expression was observed for up to 14 days after injection. The infiltration of macrophages, observed 2 days after injection in the area of GFP transduction, had dissipated by day 8 after injection. No alteration in ERG responses was observed 6 weeks after injection of rBV-CMV-GFP. CONCLUSIONS: BV efficiently transduces cultured RPE cells and many cell types in vivo in the eye, including endothelial, epithelial, and neuronal cells. BV may be a useful vector for transferring genes in cultured cells and in vivo into ocular tissue. PMID- 11726637 TI - Light-induced migration of retinal microglia into the subretinal space. AB - PURPOSE: To explore the effects of light exposure and deprivation on the distribution and function of microglia in the subretinal space of mice. METHODS: Using a monoclonal antibody, 5D4, that identifies resting, ramified microglia, the distribution and density of microglia in the retina, and the subretinal space were determined by confocal microscopy and by immunohistochemistry of cryopreserved sections of eyes of albino and pigmented mice exposed to diverse levels of light, ranging from complete darkness to intense brightness. Axotomized retinal ganglion cells were retrograde labeled by fluorescent tracer to determine whether the marker colocalizes to 5D4+ cells. Electron microscopy was used to evaluate microglia for evidence of phagocytosis. RESULTS: 5D4+ microglia in pigmented eyes were limited to the inner retinal layers, but in albino eyes 5D4+ cells were found in the outer retinal layers and subretinal space as well. The subretinal space of eyes of albino mice raised from birth in complete darkness contained few 5D4+ cells, but exposure to light caused the rapid accumulation of 5D4+ cells at this site. 5D4+ cell density in the subretinal space correlated directly with intensity of ambient light. Retrograde labeling of axotomized ganglion cells resulted in 5D4+ cells in the subretinal space that contained the retrograde label. Subretinal microglia contained phagocytized rod outer segment discs. On intense light exposure, 5D4+ cells adopted an active morphology, but failed to express class II major histocompatibility complex (MHC) molecules. CONCLUSIONS: Light exposure induced retinal microglia migration into the subretinal space in albino mice. Subretinal microglia appeared to augment through phagocytosis the capacity of pigment epithelium to take up the photoreceptor debris of light toxicity. The unexpected presence of these cells in the subretinal space raises questions concerning their potential contribution to immune privilege in this space and to the fate of retinal transplants. PMID- 11726638 TI - The fate of heterotopically grafted neural precursor cells in the normal and dystrophic adult mouse retina. AB - PURPOSE: To study the integration and differentiation of heterotopically transplanted neural precursor cells in the retina of adult mouse mutants displaying apoptotic degeneration of photoreceptor cells. METHODS: Neural precursor cells were isolated from the spinal cord of transgenic mouse embryos ubiquitously expressing enhanced green fluorescent protein. Cells were expanded in vitro and transplanted into the retina of adult wild-type and age-matched beta2/beta1 knock-in mice. Beta2/beta1 knock-in mutants display apoptotic death of photoreceptor cells and were generated by placing the cDNA of the beta1 subunit into the gene of the beta2 subunit of Na,K-ATPase. The integration and differentiation of grafted cells in recipient retinas was studied 1 or 6 months after transplantation. RESULTS: Mutant retinas contained more donor-derived cells than wild-type hosts. Moreover, in mutants, donor cells integrated into deeper retinal layers. In both genotypes, grafted cells differentiated into astrocytes and oligodendrocytes. Only a few ganglion cell axons were myelinated by donor derived oligodendrocytes 1 month after transplantation, whereas extensive myelination of the nerve fiber layer was observed 6 months after transplantation. Unequivocal evidence for differentiation of grafted cells into neurons was not obtained. CONCLUSIONS: Heterotopically transplanted neural precursor cells are capable of integrating, surviving, and differentiating into neural cell types in normal and dystrophic retinas of adult mice. The particular environment of a pathologically altered retina facilitates integration of transplanted precursor cells. In principle, neural precursors may thus be useful to substitute for or replace dysfunctional or degenerated cell types. Results of the present study also indicate that replacement of retinal cell types is likely to require more appropriate donor cells, such as retinal precursor cells. PMID- 11726639 TI - Adenosine receptor antagonists and retinal neovascularization in vivo. AB - PURPOSE: The role of adenosine receptor (AdoR) antagonists in human retinal endothelial cell function in vitro has previously been determined. In this study, efficacy of AdoR antagonist administration in reducing retinal neovascularization was examined in a mouse pup model of oxygen-induced retinopathy. METHODS: A previously described model of oxygen-induced retinal neovascularization in newborn mouse pups was used to examine the effect of various AdoR antagonists on neovascularization. The nonselective AdoR antagonist xanthine amine congener (XAC), the A(2A)-selective antagonist ZM241385, the A(2B)-selective antagonists 3 N-propylxanthine (enprofylline) and 3-isobutyl-8-pyrrolidinoxanthine (IPDX), and the A(1)-selective antagonist cyclopentyl-1,3-dipropylxanthine (CPX) were used. After the hyperoxia exposure the animals received daily intraperitoneal injections of pharmacologically relevant doses of AdoR antagonists for 5 days. Control animals received vehicle (0.1% dimethyl sulfoxide [DMSO]) alone. The animals were then killed and perfused with fluorescein-dextran. Wholemounts of retinas from one eye were prepared and examined, whereas the retinas of the contralateral eye were embedded, sectioned, and stained for counting neovascular nuclei extending beyond the internal limiting membrane into the vitreous. RESULTS: Angiography of wholemount retinas showed reduction of neovascular tufts in animals treated with selective A(2B) AdoR antagonists. Quantification of the extraretinal neovascular nuclei showed that only animals treated with XAC, enprofylline, or IPDX showed a significant reduction in retinal neovascularization. By contrast, neither CPX nor ZM241385 had an effect on neovascularization. CONCLUSIONS: The A(2B)-selective AdoR antagonists inhibited oxygen-induced retinal neovascularization in vivo and may provide a basis for developing pharmacologic therapies for the treatment of proliferative retinopathies. PMID- 11726640 TI - Angiostatin inhibits pathological but not physiological retinal angiogenesis. AB - PURPOSE: Antiangiogenic treatment is a promising new therapy for angiogenesis dependent diseases. In the current study, the biologic effects on pathologic and physiological angiogenesis in the retina of angiostatin, a very potent angiogenesis inhibitor were determined. In addition, the effects of angiostatin on the growth and development of newborn mice were examined. METHODS: Oxygen induced retinopathy was induced by subjecting mice postnatal day (P)7 to hyperoxic conditions (5 days) followed by normoxic conditions (relative hypoxia). Mice were treated with angiostatin (intravitreal or systemic). Retinal blood vessels were visualized by fluorescein angiography. Retinal neovascularization was assessed by counting intravitreal endothelial cell nuclei. Growth and organogenesis were determined between P0 and P14. RESULTS: Relative hypoxia resulted in intravitreal proliferation of retinal blood vessels. However, proliferation was inhibited completely by systemic administration of angiostatin without affecting normal retinal vascularization. After intravitreal injection of angiostatin, pathologic proliferation of the retinal blood vessels was impaired by 62%. Neither systemic nor intravitreal treatment impaired the development or growth of organs throughout the body. CONCLUSIONS: Angiostatin inhibits oxygen induced intravitreal pathologic retinal angiogenesis without affecting the development of physiological retinal vascularization, development, and growth of newborn mice. Therefore, antiangiogenic treatment may be a useful tool in the treatment of proliferative retinopathies. PMID- 11726641 TI - Diverse macular dystrophy phenotype caused by a novel complex mutation in the ELOVL4 gene. AB - PURPOSE: A 5-bp deletion in ELOVL4, a photoreceptor-specific gene, has been associated with autosomal dominant (ad) macular dystrophy phenotypes in five related families, in which phenotypes range from Stargardt-like macular dystrophy (STGD3; Mendelian Inheritance in Man 600110) to pattern dystrophy. This has been the only mutation identified in ELOVL4 to date, which is associated with macular dystrophy phenotypes. In the current study, the potential involvement was investigated of an ELOVL4 gene variation in adSTGD-like and other macular dystrophy phenotypes segregating in a large unrelated pedigree from Utah (K4175). METHODS: The entire open reading frame of the ELOVL4 gene was analyzed by direct sequencing in a proband from the K4175 family. The combination of denaturing high performance liquid chromatography (DHPLC) analysis and direct sequencing of all available family members was used to further assess segregation of identified ELOVL4 variants in the pedigree. RESULTS: A complex mutation, two 1-bp deletions separated by four nucleotides, was detected in all affected members of the family. The mutation results in a frameshift and the truncation of the ELOVL4 protein, similar to the effect of the previously described 5-bp deletion. CONCLUSIONS: The discovery of a second mutation in the ELOVL4 gene segregating with macular dystrophy phenotypes confirms the role of this gene in a subset of dominant macular dystrophies with a wide range of clinical expressions and suggests a role for modifying genes and/or environmental factors in the disease process. PMID- 11726642 TI - Transpupillary thermotherapy for subfoveal occult choroidal neovascularization: effect on ocular perfusion. AB - PURPOSE: To perform a descriptive analysis of the effects on ocular blood flow of transpupillary thermotherapy (TTT) for occult subfoveal choroidal neovascular membranes (CNVMs) in age-related macular degeneration (AMD). METHODS: Eleven subjects with occult subfoveal CNVM due to AMD were assessed in a masked fashion by color Doppler imaging (CDI) within 24 hours before, 24 hours after, and 1 month after undergoing TTT. RESULTS: In the posterior ciliary arteries (PCAs), there were no statistically significant changes observed in the peak systolic velocity (PSV), end diastolic velocity (EDV), or resistive index (RI) at 24 hours. At 1 month, the mean EDV decreased 36% (P = 0.0105) and the mean RI increased 3.8% (P = 0.0305) in the nasal PCA. Although there was a similar trend in the temporal PCA, the differences did not reach statistical significance. In the central retinal artery (CRA), the mean PSV decreased 16% (P = 0.0137), and the mean EDV decreased 21% (P = 0.0222) at 24 hours after treatment. There were no statistically significant differences in the CRA blood flow indices at 1 month after treatment. In the ophthalmic artery, there were no statistically significant differences observed in the mean PSV, EDV, or RI at 24 hours or 1 month after treatment. CONCLUSIONS: TTT is associated with transiently decreased volumetric blood flow in the retinal circulation 24 hours after treatment. In the posterior ciliary arteries that supply the choroid, there were no changes observed at 24 hours, but at 1 month, there was a decrease in the mean EDV and an increase in the RI in the nasal and temporal PCAs, reaching statistical significance in the nasal PCA only. This study suggests that TTT could lead to alterations in choroidal blood flow, as assessed by CDI. Further study is warranted. PMID- 11726643 TI - Mapping of glaucomatous visual field defects by multifocal VEPs. AB - PURPOSE: To objectively measure a visual field and to sensitively detect glaucomatous visual field defects by measuring the contrast sensitivity function (CSF), using multifocal visual evoked cortical potentials (MVEPs). METHODS: MVEPs of normal subjects (n = 28) and of patients with glaucoma (n = 12) or ocular hypertension (OH, n = 1) were recorded. A multi-input procedure was used to obtain 37 local VEP responses to each scaled hexagon, composed of 24 triangular patterns, reversing in a counterphase manner. Two pattern contrasts of 32% and 8% were used for measuring the contrast threshold. To improve the signal-to-noise ratio, 37 MVEPs were averaged into 20 groups. The root-mean-square (RMS) measures at both contrasts were calculated. Contrast thresholds were estimated by extrapolating the regression line of the amplitude versus contrast to the mean noise levels. RESULTS: RMS amplitudes of each local MVEP decreased as the eccentricity increased and as the pattern contrast decreased in normal subjects. It was also revealed that the amplitudes were smaller in the upper-half field than those in the lower-half field. Compared with the RMS amplitudes, CSFs estimated by MVEPs were relatively constant without being strongly influenced by retinal eccentricity. In patients with glaucoma, the CSFs, even from the locations where the mean perimetric sensitivities ranged to more than 30 dB, were significantly smaller than those in the normal control subjects (P < 0.001, Mann Whitney test). CSFs in the 20- to 30-dB and 100 to 20-dB groups were also significantly smaller than those in the more-than-30-dB or 20- to 30-dB groups, respectively (P < 0.001, Mann-Whitney test). A significant correlation was found between the logarithmic function of the CSF and perimetric sensitivity (r = 0.57, P < 0.001, n = 216). The CSFs were evaluated on the basis of normal SD. Mappings of the CSFs agreed well with those of the perimetric sensitivity in all patients with glaucoma and was even more sensitive in detecting slight optic nerve damage by glaucoma than was perimetry. CONCLUSIONS: Local optic nerve damage caused by glaucoma can be sensitively detected by measuring contrast sensitivity using the MVEP. PMID- 11726644 TI - Optical response to LASIK surgery for myopia from total and corneal aberration measurements. AB - PURPOSE: To evaluate the optical aberrations induced by LASIK refractive surgery for myopia on the anterior surface of the cornea and the entire optical system of the eye. METHODS: Total and corneal aberrations were measured in a group of 14 eyes (preoperative myopia ranging from -2.5 to -13 D) before and after LASIK surgery. Total aberrations were measured using a laser ray-tracing technique. Corneal aberrations were obtained from corneal elevation maps measured using a corneal system and custom software. Corneal and total wave aberrations were described as Zernike polynomial expansions. Root-mean-square (RMS) wavefront error was used as a global optical quality metric. RESULTS: Total and corneal aberrations (third-order and higher) showed a statistically significant increase after LASIK myopia surgery, by a factor of 1.92 (total) and 3.72 (corneal), on average. This increase was more pronounced in patients with the highest preoperative myopia. There is a good correlation (r = 0.97, P < 0.0001) between the aberrations induced in the entire optical system and those induced in the anterior corneal surface. However, the anterior corneal spherical aberration increased more than the total spherical aberration, suggesting also a change in the spherical aberration of the posterior corneal surface. Pupil centration and internal optical aberrations, which are not accounted for in corneal topography, play an important role in evaluating individual surgical outcomes. CONCLUSIONS: Because LASIK surgery induces changes in the anterior corneal surface, most changes in the total aberration pattern can be attributed to changes in the anterior corneal aberrations. However, because of individual interactions of the aberrations in the ocular components, a combination of corneal and total aberration measurements is critical to understanding individual outcomes, and by extension, to designing custom ablation algorithms. This comparison also reveals changes in the internal aberrations, consistent with the posterior corneal changes reported using scanning slit corneal topography. PMID- 11726645 TI - Is color vision deficiency an advantage under scotopic conditions? AB - PURPOSE: To examine experimentally whether color vision deficiency confers a selective advantage under scotopic conditions. METHODS: Red-green color-deficient subjects, monochromats, and age-matched color-normal control subjects were examined. In each subject the time course of dark adaptation, scotopic visual field sensitivity, and performance on a scotopic perceptual task were measured. RESULTS: No significant differences were found between red-green color-deficient subjects and control subjects on any of the three tests. Our small sample of monochromats had higher absolute thresholds than the corresponding control subjects, but their performance at the scotopic visual field test and perceptual task did not differ significantly from that of color-normal subjects. CONCLUSIONS: No evidence was found that red-green color deficiency or monochromatism confers a selective advantage under scotopic conditions. PMID- 11726646 TI - Protective roles of NF-kappa B for chromium(VI)-induced cytotoxicity is revealed by expression of Ikappa B kinase-beta mutant. AB - To delineate the molecular mechanisms of NF-kappaB-mediated regulation of chromium(VI)-induced cell death, the signaling pathway leading to the activation of NF-kappaB was interrupted by stable transfection of a kinase-mutated form of IkappaB kinase beta (IKKbeta-KM). Here we demonstrate a novel role for the NF kappaB transcription factor in inhibiting chromium(VI)-induced cell death. Inhibition of NF-kappaB by IKKbeta-KM or IKKbeta gene deficiency resulted in a spontaneous cleavage of Bcl-xL antiapoptotic protein due to the elevated caspase 3 activity. DNA microarray assay suggested a decreased expression of genes encoding antiapoptotic proteins, cIAP1 and cIAP2, in the cells overexpressing IKKbeta-KM. Chromium(VI) treatment of these NF-kappaB-inhibited cells induced necrotic-like cell death. Such chromium(VI)-induced cell killing could be partially inhibited by expression of exogenous cIAP1, an inhibitor of caspases, indicating that caspases along with others may be involved in chromium(VI) induced cell death. These results suggest that NF-kappaB is essential for inhibiting toxic metal-induced cytotoxicity. Such inhibition may involve up regulation of the expression of anti-death proteins including cIAP1 that prevents spontaneous caspase activation and subsequent cleavage of Bcl-xL protein. PMID- 11726647 TI - Prolonged nuclear retention of activated extracellular signal-regulated protein kinase promotes cell death generated by oxidative toxicity or proteasome inhibition in a neuronal cell line. AB - In the HT22 mouse hippocampal cell line and primary immature embryonic rat cortical neurons, glutamate-induced oxidative toxicity is associated with a delayed but chronic activation of extracellular signal-regulated kinase-1/2 (ERK 1/2). ERK-1/2 is also activated in HT22 cells that undergo caspase-dependent cell death upon inhibition of proteasome-dependent protein degradation brought about by MG132 treatment. As in glutamate-treated HT22 cells and primary neurons, inhibition of MEK-1, an upstream activator of ERK-1/2 protects against MG132 induced toxicity. Furthermore, activated ERK-1/2 is retained within the nucleus in glutamate- and MG132-treated HT22 cells. Although previous studies suggested that ERK-1/2 activation was downstream of many cell death-inducing signals in HT22 cells, we show here that cycloheximide, the Z-vad caspase inhibitor, and a nonlethal heat shock protect against glutamate- and MG132-induced toxicity without diminishing ERK-1/2 activation. In these cases, ERK-1/2, although chronically activated, is not retained within the nucleus but accumulates within the cytoplasm. Thus, persistent nuclear retention of activated ERK-1/2 may be a critical factor in eliciting proapoptotic effects in neuronal cells subjected to oxidative stress or proteasome inhibition. PMID- 11726648 TI - Interleukin (IL)-4 indirectly suppresses IL-2 production by human T lymphocytes via peroxisome proliferator-activated receptor gamma activated by macrophage derived 12/15-lipoxygenase ligands. AB - The respective development of either T helper type 1 (Th1) or Th2 cells is believed to be mediated by the effects of cytokines acting directly on Th precursors (Thp). We have generated evidence for an indirect monocyte-dependent immunoregulatory pathway. Recently, interleukin (IL) 4 has been shown to produce "new" potential peroxisome proliferator-activated receptor gamma (PPARgamma) ligands by inducing macrophage 12/15-lipoxygenase (12/15-LO). We have shown previously that the activated PPARgamma is a profound inhibitor of IL-2 transcription in human T lymphocytes. It is hypothetically possible that IL-4 might indirectly affect IL-2 production by Thp cells via macrophage-derived PPARgamma ligands. Using human monocytes and T lymphocytes from same donors, we have found that monocyte 12/15-LO products mediate the indirect inhibitory effect of IL-4 on anti-CD3- or phytohemagglutinin/phorbol 12-myristate 13-acetate stimulated IL-2 production by T lymphocytes. We further analyzed which major 12/15-LO metabolites contributed to the above inhibition. 13 Hydroxyoctadecadienoic acid (13-HODE), a 12/15-LO product, markedly blocked IL-2 production by human blood T lymphocytes, but not Jurkat T cells. Moreover, the IL 4-conditioned macrophage medium contained a sufficient amount of 13-HODE and anti 13-HODE antibody indeed neutralized the inhibitory effects of the IL-4 conditional medium on T-cell IL-2 production. Using human T lymphocytes and the PPARgamma-transfected Jurkat T cells, we demonstrated the specific inhibition by 13-HODE of the transcription factors NFAT (nuclear factor of activated T cells) and nuclear factor kappaB, the IL-2 promoter reporter, and IL-2 production. However, 15-hydroxytetraenoic acid had little inhibitory effect. The potency of such inhibitory effects correlates well with the capability of the above metabolic lipids to activate PPARgamma. These data provide a mechanism whereby IL 4 may indirectly affect Thp function via PPARgamma activated by macrophage products of the 12/15-LO pathway. PMID- 11726649 TI - Distinct role of CD80 and CD86 in the regulation of the activation of B cell and B cell lymphoma. AB - To date, not much has been known regarding the role of CD80 and CD86 molecules in signaling of B cells. The CD28/CTLA4 ligands, CD80 (B7-1) and CD86 (B7-2), are expressed on the surface of freshly isolated splenic B cells, and their expression is up-regulated by lipopolysaccharides. In the present study, we have investigated whether signaling via CD80/CD86 could alter the proliferation and immunoglobulin synthesis of B cells. Splenic B cells were stimulated with lipopolysaccharides in the presence of anti-B7-1 (16-10A1) and anti-B7-2 (GL1) monoclonal antibodies (mAbs). Exciting features observed during the study were that cross-linking of CD86 with GL1 enhanced the proliferation and production of IgG1 and IgG2a isotypes. In contrast, anti-B7-1 (16-10A1) mAb could efficiently block the proliferation and production of IgG1 and IgG2a. Furthermore, GL1 mAb could also induce the secretion of IgG isotypes from B cell lymphomas. Importantly, 16-10A1 could retard the growth of lymphomas and favored the up regulation of pro-apoptotic molecules caspase-3, caspase-8, Fas, FasL, Bak, and Bax and down-regulation of anti-apoptotic molecule Bcl-x(L). In contrast, GL1 augmented the level of anti-apoptotic molecules Bcl-w and Bcl-x(L) and decreased the levels of pro-apoptotic molecule caspase-8, thereby providing a novel insight into the mechanism whereby triggering through CD80 and CD86 could deliver regulatory signals. Thus, this study is the first demonstration of a distinct signaling event induced by CD80 and CD86 molecules in B cell lymphoma. Finally, the significance of the finding is that CD80 provided negative signal for the proliferation and IgG secretion of normal B cells and B cell lymphomas. In contrast, CD86 encouraged the activity of B cells. PMID- 11726650 TI - Mutation of the matrix metalloproteinase At2-MMP inhibits growth and causes late flowering and early senescence in Arabidopsis. AB - This study characterizes the expression and functional significance of the member of the matrix metalloproteinase (MMP) family At2-MMP from Arabidopsis. By transcript analysis, expression of At2-MMP was found in leaves and roots of juvenile Arabidopsis and leaves, roots, and inflorescences of mature flowering plants showing strong increase of transcript abundance with aging. Cell specificity of expression of At2-MMP was studied by in situ hybridizations in leaves and flowers of Arabidopsis. In leaves, the gene was expressed in the phloem, in developing xylem elements, epidermal cells, and neighboring mesophyll cell layers. In flowers, signals were localized in pistils, ovules, and receptacles. In an Arabidopsis mutant (at2-mmp-1) carrying a tDNA insertion in At2-MMP, neither germination nor development of plants was modified in comparison to the wild type in the juvenile rosette stage. Starting with the onset of shoots, growth of roots, leaves, and shoots was inhibited compared with the wild type, and the plants were characterized by late flowering. Besides the flowering, at2-mmp-1 plants showed fast degradation of chlorophyll in leaves and early senescence. These results demonstrate the involvement of At2-MMP in plant growth, morphogenesis, and development with particular relevance for flowering and senescence. PMID- 11726651 TI - Identification of a region of RyR1 that participates in allosteric coupling with the alpha(1S) (Ca(V)1.1) II-III loop. AB - In skeletal muscle, excitation-contraction (EC) coupling and retrograde signaling are thought to result from direct interactions between the ryanodine receptor (RyR1) and the alpha(1) subunit of the dihydropyridine receptor (alpha(1S)). Previous work has shown that the s53 region of alpha(1S) (residues 720-765 in the II-III loop) and regions R10 (1635-2636) and R9 (2659-3720) of RyR1 are involved in this signaling. Using the yeast two-hybrid system, we here report an interaction between s53 and the sR16 region of RyR1 (1837-2168, within R10), whereas no interaction was seen using upstream residues of the alpha(1S) II-III loop (s31, 666-709). The specificity of the s53-sR16 interaction was tested by using fragments of the cardiac RyR (RyR2) and DHPR (alpha(1C)) that correspond to sR16 and s53, respectively. No interaction was observed for sR16 x c53 (alpha(1C) 850-897), but weak interaction was occasionally observed for s53 x cR16 (RyR2 1817-2142). To test the functional significance of the s53 x sR16 interaction, we expressed in dyspedic myotubes a chimeric RyR (chimeraR16) in which sR16 was substituted for the corresponding region of RyR2. ChimeraR16 was found to mediate weak skeletal-type EC coupling. To test the necessity of sR16 sequence for coupling, we used "chimeraR16-rev," in which sR16 and a small upstream region of RyR1 were replaced by RyR2 sequence. ChimeraR16-rev did not differ from RyR1 in its ability to mediate EC coupling. Thus, interaction between residues 720-765 of alpha(1S) and residues 1837-2168 of RyR1 appears to contribute to but is not essential for EC coupling in skeletal muscle. PMID- 11726652 TI - Inhibition of insulin receptor catalytic activity by the molecular adapter Grb14. AB - Grb14 belongs to the Grb7 family of adapters and was recently identified as a partner of the insulin receptor (IR). Here we show that Grb14 inhibits in vitro IR substrate phosphorylation. Grb14 does not alter the K(m) for ATP and behaves as an uncompetitive inhibitor for the IR substrate. Similar experiments performed with other members of the Grb7 family, Grb7 and Grb10, and with IGF-1 receptor argue in favor of a specific inhibition of the IR catalytic activity by Grb14. The IR-interacting domain of Grb14, the PIR, is sufficient for the inhibitory effect of Grb14, whereas the SH2 domain has no effect on IR catalytic activity. In Chinese hamster ovary (CHO) cells overexpressing both IR and Grb14, Grb14 binds to the IR as early as 1 min after insulin stimulation, and the two proteins remain associated. When interacting with Grb14, the IR is protected against tyrosine phosphatases action and therefore maintained under a phosphorylated state. However, the binding of Grb14 to the IR induces an early delay in the activation of Akt and ERK1/2 in CHO-IR cells, and ERK1/2 are less efficiently phosphorylated. These findings show that Grb14 is a direct inhibitor of the IR catalytic activity and could be considered as a modulator of insulin signaling. PMID- 11726653 TI - Specificities of N-acetylglucosamine-6-O-sulfotransferases in relation to L selectin ligand synthesis and tumor-associated enzyme expression. AB - N-Acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) catalyzes the transfer of sulfate from adenosine 3'-phosphate,5'-phosphosulfate to the C-6 position of the non-reducing GlcNAc. Three human GlcNAc6STs, namely GlcNAc6ST-1, GlcNAc6ST-2 (HEC GlcNAc6ST), and GlcNAc6ST-3 (I-GlcNAc6ST), were produced as fusion proteins to protein A, and their substrate specificities as well as their enzymological properties were determined. Both GlcNAc6ST-1 and GlcNAc6ST-2 efficiently utilized the following oligosaccharide structures as acceptors: GlcNAcbeta1-6[Galbeta1 3]GalNAc-pNP (core 2), GlcNAcbeta1-6ManOMe, and GlcNAcbeta1-2Man. The ratios of activities to these substrates were not significantly different between the two enzymes. However, GlcNAc6ST-2 but not GlcNAc6ST-1 acted on core 3 of GlcNAcbeta1 3GalNAc-pNP. GlcNAc6ST-3 used only the core 2 structure among the above mentioned oligosaccharide structures. The ability of GlcNAc6ST-1 to sulfate core 2 structure as efficiently as GlcNAc6ST-2 is consistent with the view that GlcNAc6ST-1 is also involved in the synthesis of l-selectin ligand. Indeed, cells doubly transfected with GlcNAc6ST-1 and fucosyltransferase VII cDNAs supported the rolling of L-selectin-expressing cells. The activity of GlcNAc6ST-2 on core 3 and its expression in mucinous adenocarcinoma suggested that this enzyme corresponds to the sulfotransferase, which is specifically expressed in mucinous adenocarcinoma (Seko, A., Sumiya, J., Yonezawa, S., Nagata, K., and Yamashita, K. (2000) Glycobiology 10, 919-929). PMID- 11726654 TI - Nuclear receptor peroxisome proliferator-activated receptor alpha (PPARalpha) is expressed in resting murine lymphocytes. The PPARalpha in T and B lymphocytes is both transactivation and transrepression competent. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors that belong to the nuclear hormone receptor superfamily. PPARalpha and PPARgamma ligands have been demonstrated to exert anti-inflammatory activities in macrophages by repressing the activities of several transcription factors. PPARgamma is expressed in T lymphocytes and may play a role in cytokine production, cellular proliferation, and susceptibility to apoptosis. Herein, we demonstrate that T and B lymphocytes constitutively express PPARalpha. PPARalpha represents the predominant isoform expressed in lymphocytes, whereas PPARgamma dominates in all cell types of the myeloid lineage. PPARalpha expression was down regulated following T-cell activation while PPARgamma expression increased under the same activating conditions. PPARalpha expression in T cells may be regulated by microenvironmental factors, because Peyer's patch T cells expressed far greater levels of PPARalpha than T cells isolated from peripheral lymphoid organs. Exposure to specific ligand determined that PPARalpha in lymphocytes can effectively transactivate a peroxisome proliferator response element reporter construct. PPARalpha's ability to regulate endogenous genes, however, required treatment with histone deacetylase inhibitors. Finally, ligand activation of lymphocyte PPARalpha antagonized NF-kappaB. Our observation that a functional PPARalpha exists within T cells and B lymphocytes suggests an expanding role for this nuclear receptor in cells of the immune system. PMID- 11726655 TI - Residues 88-109 of factor IXa are important for assembly of the factor X activating complex. AB - Activated platelets and phospholipid vesicles promote assembly of the intrinsic factor X (FX) activating complex by presenting high-affinity binding sites for blood coagulation FIXa, FVIIIa, and FX. Previous reports suggest that the second epidermal growth factor (EGF)-like domain of FIXa mediates assembly of the FX activating complex (Ahmad, S. S., Rawala, R., Cheung, W. F., Stafford, D. W., and Walsh, P. N. (1995) Biochem. J. 310, 427-431; Wong, M. Y., Gurr, J. A., and Walsh, P. N. (1999) Biochemistry 38, 8948-8960). To identify important residues, we prepared several chimeric FIXa proteins using homologous sequences from FVII: FIXa(FVIIEGF2) (FIX Delta 88-124,inverted Delta FVII91-127), FIXa(loop1) (FIX Delta 88-99,inverted Delta FVII91-102), FIXa(loop2) (FIX Delta 95-109,inverted Delta FVII98-112), FIXa(loop3) (FIX Delta 111-124,inverted Delta FVII114-127), and point mutants (FIXaR94D and FIXa(loop1)G94R). In the presence and absence of FVIIIa, a 2- to 10-fold reduced V(max) of FX activation (nm FXa min(-1)) was observed for FIXa(FVIIEGF2), FIXa(loop1), FIXa(loop2), and FIXa(loop1)G94R, whereas FIXa(loop3) and FIXaR94D were normal. For all of the FIXa proteins, K(m)((app)) values were normal as were EC(50) values for interactions with FVIIIa. However, K(d)((app)) (in nm) for the FX activating complex assembled on phospholipid vesicles was increased for FIXa(FVIIEGF2) (43.3 +/- 2.70), FIXa(loop1)(10.9 +/- 2.8), FIXa(loop2) (70.5 +/- 1.60), and FIXa(loop1)G94R (17.1 +/- 2.90) relative to FIXa(N) (3.9 +/- 0.11), FIXa(WT) (4.6 +/- 0.17), FIXa(loop3) (4.5 +/- 0.20), and FIXaR94D (2.2 +/- 0.09) suggesting that reduced V(max) is a result of impaired complex assembly. These data indicate that residues 88-109 (but not Arg(94)) are important for normal assembly of the FX activating complex on phospholipid vesicles. PMID- 11726657 TI - HIV Nef increases T cell ERK MAP kinase activity. AB - The human immunodeficiency regulatory protein Nef enhances viral replication and is central to viral pathogenesis. Although Nef has displayed a capacity to associate with a diverse assortment of cellular molecules and to increase T cell activity, the biochemical activity of Nef in T cells remains poorly defined. In this report we examine the bioactivity of Nef in primary CD4 T cells and, in particular, focus on the biochemical pathways known to be central to T cell activity. The extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase pathway was dramatically affected by Nef expression with increases in ERK, MEK, and Elk induction. The capacity of Nef to increase the MAP kinase pathway activity was dependent on T cell receptor stimulation. By increasing ERK MAP kinase activity, Nef is functionally associated with a kinase known to affect T cell activity, viral replication, and viral infectivity. PMID- 11726656 TI - Role of JunB in erythroid differentiation. AB - The role of junB as a regulator of erythroid cell survival, proliferation, and differentiation was tested by controlled expression of JunB in the erythropoietin (EPO)-dependent erythroleukemia cell line HCD57. JunB induced erythroid differentiation as evidenced by increased expression of the erythroid-specific proteins beta-globin, spectrin-alpha, and TER-119. Expression of JunB for at least 48 h was required for the differentiated phenotype to emerge. Differentiation was accompanied by a slower rate of proliferation and an increase in the expression of the cell cycle inhibitory protein p27. p27 protein expression increased due to reduced turnover without changes in transcription, indicating global changes in cell physiology following JunB induction. JunB expression was also studied in mouse and human primary erythroid cells. JunB expression increased immediately in both primary mouse cells and HCD57 cells treated with EPO and quickly returned to base-line levels, followed by a secondary rise in JunB in primary erythroid cells, but not in HCD57 cells, 36-48 h later. This result suggested that the initial EPO-dependent JunB induction was not sufficient to induce differentiation, but that the late EPO-independent JunB expression in primary erythroid cells was necessary for differentiation. This study suggests that JunB is an important regulator of erythroid differentiation. PMID- 11726658 TI - Enzymatically active paraoxonase-1 is located at the external membrane of producing cells and released by a high affinity, saturable, desorption mechanism. AB - Paraoxonase-1 (PON1) is a high density lipoprotein (HDL)-associated serum enzyme that protects low density lipoproteins from oxidative modifications. There is a relative lack of information on mechanisms implicated in PON1 release from cells. The present study focused on a model derived from stable transfection of CHO cells, to avoid co-secretion of apolipoprotein (apo) A-I and lipids, which could lead to formation of HDL-like complexes. Our results indicate that, in the absence of an appropriate acceptor, little PON1 is released. The results designate HDL as the predominant, physiological acceptor, whose efficiency is influenced by size and composition. Neither lipid-poor apoA-I or apoA-II nor low density lipoproteins could substitute for HDL. Protein-free phospholipid complexes promoted PON1 release. However, the presence of both apolipoprotein and phospholipid were necessary to promote release and stabilize the enzyme. Immunofluorescence studies demonstrated that PON1 was inserted into the external membrane of CHO cells, where it was enzymatically active. Accumulation of PON1 in the cell membrane was not influenced by the ability of the cell to co-secrete of apoA-I. Release appeared to involve desorption by HDL; human and reconstituted HDL promoted PON1 release in a saturable, high affinity manner (apparent affinity 1.59 +/- 0.3 microg of HDL protein/ml). Studies with PON1-transfected hepatocytes (HuH-7) revealed comparable structural features with the peptide located in a punctate pattern at the external membrane and enzymatically active. We hypothesize that release of PON1 involves a docking process whereby HDL transiently associate with the cell membrane and remove the peptide from the external membrane. The secretory process may be of importance for assuring the correct lipoprotein destination of PON1 and thus its functional efficiency. PMID- 11726659 TI - Novel protein modification by kynurenine in human lenses. AB - It is known that human lenses increase in color and fluorescence with age, but the molecular basis for this is not well understood. We demonstrate here that proteins isolated from human lenses contain significant levels of the UV filter kynurenine covalently bound to histidine and lysine residues. Identification was confirmed by synthesis of the kynurenine amino acid adducts and comparison of the chromatographic retention times and mass spectra of these authentic standards with those of corresponding adducts isolated from human lenses following acid hydrolysis. Using calf lens proteins as a model, covalent binding of kynurenine to lens proteins has been shown to proceed via side chain deamination in a manner analogous to that observed for the related UV filter, 3-hydroxykynurenine O-beta D-glucoside. Levels of histidylkynurenine and lysylkynurenine were low in human lenses in subjects younger than 30, but thereafter increased in concentration with the age of the individual. Post-translational modification of lens proteins by tryptophan metabolites therefore appears to be responsible, at least in part, for the age-dependent increase in coloration and fluorescence of the human lens, and this process may also be important in other tissues in which up-regulation of tryptophan catabolism occurs. PMID- 11726660 TI - Insulin-like growth factor II plays a central role in atherosclerosis in a mouse model. AB - Insulin-like growth factor II is a fetal promoter of cell proliferation that is involved in some forms of cancer and overgrowth syndromes in humans. Here, we provide two sources of genetic evidence for a novel, pivotal role of locally produced insulin-like growth factor II in the development of atherosclerosis. First, we show that homozygosity for a disrupted insulin-like growth factor II allele in mice lacking apolipoprotein E, a widely used animal model of atherosclerosis, results in aortic lesions that are approximately 80% smaller and contain approximately 50% less proliferating cells compared with mice lacking only apolipoprotein E. Second, targeted expression of an insulin-like growth factor II transgene in smooth muscle cells, but not the mere elevation of circulating levels of the peptide, causes per se aortic focal intimal thickenings. The insulin-like growth factor II transgenics presented here are the first viable mutant mice spontaneously developing intimal masses. These observations provide the first direct evidence for an atherogenic activity of insulin-like growth factor II in vivo. PMID- 11726661 TI - Osteoprotegerin ligand induces beta-casein gene expression through the transcription factor CCAAT/enhancer-binding protein beta. AB - Osteoprotegerin ligand (OPGL, also known as RANKL), a member of the tumor necrosis factor superfamily, is essential for mammary gland development during pregnancy in addition to key roles in the immune system and bone development. Here we show that OPGL induces beta-casein transcription through the CCAAT/enhancer-binding protein beta (C/EBPbeta). In both HC11 cell lines and primary mammary epithelial cells, OPGL stimulation triggers rapid nuclear translocation of C/EBPbeta, which is critical for the expression of the beta casein gene. Mutation of C/EBbeta binding sites in the beta-casein gene promoter completely abrogated OPGL-induced beta-casein promoter activity. By contrast, OPGL stimulation did not result in STAT5 phosphorylation. In vivo immunohistochemistry studies further demonstrated defective nuclear translocation of C/EBPbeta, but normal STAT5 activation, in OPGL-deficient mice. These data show that OPGL is a critical activator of beta-casein gene expression via the transcription factor C/EBPbeta. Our data provide new insights into the understanding of the molecular events involved in milk protein gene expression. PMID- 11726662 TI - Heparan sulfate modulates kinin release by Trypanosoma cruzi through the activity of cruzipain. AB - Trypanosoma cruzi activates the kinin pathway through the activity of its major cysteine proteinase, cruzipain. Because kininogen molecules may be displayed on cell surfaces by binding to glycosaminoglycans, we examined whether the ability of cruzipain to release kinins from high molecular weight kininogen (HK) is modulated by heparan sulfate (HS). Kinetic assays show that HS reduces the cysteine proteinase inhibitory activity (K(i app)) of HK about 10-fold. Conversely, the catalytic efficiency of cruzipain on kinin-related synthetic fluorogenic substrates is enhanced up to 6-fold in the presence of HS. Analysis of the HK breakdown products generated by cruzipain indicated that HS changes the pattern of HK cleavage products. Direct measurements of bradykinin demonstrated an up to 35-fold increase in cruzipain-mediated kinin liberation in the presence of HS. Similarly, kinin release by living trypomastigotes increased up to 10-fold in the presence of HS. These studies suggest that the efficiency of T. cruzi to initiate kinin release is potently enhanced by the mutual interactions between cruzipain, HK, and heparan sulfate proteoglycans. PMID- 11726663 TI - DNA damage invokes mismatch repair-dependent cyclin D1 attenuation and retinoblastoma signaling pathways to inhibit CDK2. AB - DNA-damage evokes cell cycle checkpoints, which function to maintain genomic integrity. The retinoblastoma tumor suppressor (RB) and mismatch repair complexes are known to contribute to the appropriate cellular response to specific types of DNA damage. However, the signaling pathways through which these proteins impact the cell cycle machinery have not been explicitly determined. RB-deficient murine embryo fibroblasts continued a high degree of DNA replication following the induction of cisplatin damage, but were inhibited for G(2)/M progression. This damage led to RB dephosphorylation/activation and subsequent RB-dependent attenuation of cyclin A and CDK2 activity. In both Rb+/+ and Rb -/- cells, cyclin D1 expression was attenuated following DNA damage. As cyclin D1 is a critical determinant of RB phosphorylation and cell cycle progression, we probed the pathway through which cyclin D1 degradation occurs in response to DNA damage. We found that attenuation of endogenous cyclin D1 is dependent on multiple mismatch repair proteins. We demonstrate that the mismatch repair-dependent attenuation of endogenous cyclin D1 is critical for attenuation of CDK2 activity and induction of cell cycle checkpoints. Together, these studies couple the activity of the retinoblastoma and mismatch repair tumor suppressor pathways through the degradation of cyclin D1 and dual attenuation of CDK2 activity. PMID- 11726664 TI - Intergenic mRNA molecules resulting from trans-splicing. AB - Accumulated recent evidence is indicating that alternative splicing represents a generalized process that increases the complexity of human gene expression. Here we show that mRNA production may not necessarily be limited to single genes, as human liver also has the potential to produce a variety of hybrid cytochrome P450 3A mRNA molecules. The four known cytochrome P450 3A genes in humans, CYP3A4, CYP3A5, CYP3A7, and CYP3A43, share a high degree of similarity, consist of 13 exons with conserved exon-intron boundaries, and form a cluster on chromosome 7. The chimeric CYP3A mRNA molecules described herein are characterized by CYP3A43 exon 1 joined at canonical splice sites to distinct sets of CYP3A4 or CYP3A5 exons. Because the CYP3A43 gene is in a head-to-head orientation with the CYP3A4 and CYP3A5 genes, bypassing transcriptional termination can not account for the formation of hybrid CYP3A mRNAs. Thus, the mechanism generating these molecules has to be an RNA processing event that joins exons of independent pre-mRNA molecules, i.e. trans-splicing. Using quantitative real-time polymerase chain reaction, the ratio of one CYP3A43/3A4 intergenic combination was estimated to be approximately 0.15% that of the CYP3A43 mRNAs. Moreover, trans-splicing has been found not to interfere with polyadenylation. Heterologous expression of the chimeric species composed of CYP3A43 exon 1 joined to exons 2-13 of CYP3A4 revealed catalytic activity toward testosterone. PMID- 11726665 TI - Dimeric versions of two short N-cadherin binding motifs (HAVDI and INPISG) function as N-cadherin agonists. AB - N-cadherin is a member of the classical cadherin family of homophilic binding molecules. Peptide competition studies have identified the HAVDI and INPISGQ sequences as functional binding motifs in extracellular domain 1 (ECD1) of N cadherin. Whereas monomeric versions of these motifs function as specific N cadherin antagonists, we now show that cyclic peptides containing a tandem repeat of the individual motifs function as N-cadherin agonists. In this context, when presented to neurons as soluble molecules, the dimeric versions of the motifs stimulate neurite outgrowth in a similar manner to native N-cadherin. The response to the dimeric agonist peptides was inhibited by monomeric versions of the same motif and also by recombinant N-cadherin ECD1 protein. The responses were also inhibited by antibodies to a fibroblast growth factor receptor (FGFR) binding motif in ECD4 of N-cadherin and by a specific FGFR antagonist (PD17304). These data suggest that the peptides function by binding to and clustering N cadherin in neurons and thereby activating an N-cadherin/FGFR signaling cascade. The novel agonists will be invaluable for dissecting out those cadherin functions that rely on signaling as opposed to adhesion and clearly have the potential to be developed as therapeutic agents for the promotion of cell survival and axonal regeneration. PMID- 11726666 TI - Molecular cloning and characterization of a novel histone deacetylase HDAC10. AB - The growing number of proteins controlled by reversible acetylation suggests the existence of a large number of acetyltransferases and deacetylases. Here, we report the identification of a novel class II histone deacetylase, HDAC10. Homology comparison indicates that HDAC10 is most similar to HDAC6. Both contain a unique, putative second catalytic domain not found in other HDACs. In HDAC10, however, this domain is not functional. This tandem organization of two catalytic domains confers resistance to the inhibitors trapoxin B and sodium butyrate, which potently inhibit the deacetylase activity of all other HDAC members. Thus, HDAC10 and HDAC6 share unusual structural and pharmacological characteristics. However, unlike HDAC6, which is normally a cytoplasmic deacetylase, HDAC10 resides in both the nucleus and cytoplasm. In the nucleus, when tethered to a promoter, HDAC10 represses transcription independent of its deacetylase activity, indicating that HDAC10 contains a distinct transcriptional repressor domain. These observations suggest that HDAC10 might uniquely play roles both in the nucleus, as a transcriptional modulator, and in the cytoplasm in an unidentified role. Together, our results identify HDAC10 as a novel deacetylase with distinct structure, pharmacology and localization and further expand the complexity of the HDAC family. PMID- 11726667 TI - Comparison of TATA-binding protein recognition of a variant and consensus DNA promoters. AB - Assembly of transcription pre-initiation complexes proceeds from the initial complex formed between "TATA" bearing promoter DNA and the TATA-binding protein (TBP). Our laboratory has been investigating the relationships among TATA sequence, TBP center dot TATA solution structure, recognition mechanisms, and transcription efficiency. TBP center dot TATA interactions have been modeled by global analysis of detailed kinetic and thermodynamic data obtained using fluorimetric and fluorometric techniques in conjunction with fluorescence resonance energy transfer. We have reported recently that TBP recognition of two consensus promoters, adenovirus major late (AdMLP: TATAAAAG) and E4 (TATATATA), is well described by a linear two-intermediate mechanism with simultaneous DNA binding and bending. Similar DNA geometries and high transcription efficiencies characterize these TBP x TATA complexes. Here we show that, in contrast to the consensus sequences, TBP recognition of a variant sequence (C7: TATAAACG) is described by a three-step model with two branching pathways. One pathway proceeds through an intermediate having severely bent DNA, reminiscent of the consensus interactions, with the other branch yielding a unique conformer with shallowly bent DNA. The resulting TBP x C7 complex has a dramatically different solution conformation than for TBP x DNA(CONSENSUS) and is correlated with diminished relative transcription activity. The temperature dependence of the TBP x C7 helical bend is postulated to derive from population shifts between the conformers with slightly and severely bent DNA. PMID- 11726668 TI - Internalization determinants of the parathyroid hormone receptor differentially regulate beta-arrestin/receptor association. AB - beta-Arrestins have been implicated in regulating internalization of the parathyroid hormone receptor (PTHR), but the structural features in the receptor required for this effect are unknown. In the present study performed in HEK-293 cells, we demonstrated that different topological domains of PTHR are implicated in agonist-dependent receptor internalization; truncation of the cytoplasmic tail (PTHR-TR), selective mutations of the cytoplasmic tail to remove the sites of parathyroid hormone (PTH)-stimulated phosphorylation (PTHR-PD), and mutations in the third transmembrane helix (N289A) or in the third cytoplasmic loop (K382A) resulted in a 30-60% reduction in (125)I-PTH-related protein internalization. To better define the role of these internalization determinants, we have tested the ability of these mutant PTHRs to associate with beta-arrestins by using three different methodological approaches: 1) ability of overexpression of beta arrestins to restore the internalization of (125)I-PTH-related protein for the mutant PTHRs; 2) visualization of PTH-mediated trafficking of beta-arrestin1 and 2 fused to the green fluorescent protein with receptors by confocal microscopy; 3) quantification of beta-arrestin1-green fluorescent protein translocation by Western blot. Our data reveal that the receptor' cytoplasmic tail contains determinants of beta-arrestin interaction that are distinct from the phosphorylation sites and are sufficient for transient association of beta arrestin2, but stable association requires receptor phosphorylation. Determinants in the receptor's core (Asn-289 and Lys-382) appear to regulate internalization of the receptor/beta-arrestin complex toward early endocytic endosomes during the initial step of endocytosis. PMID- 11726669 TI - The interferon (IFN)-induced GTPase, mGBP-2. Role in IFN-gamma-induced murine fibroblast proliferation. AB - To investigate the function of mGBP-2, a member of the interferon (IFN)-induced guanylate-binding protein family of GTPases, NIH 3T3 fibroblasts were generated that constitutively expressed mGBP-2. mGBP-2 induced a faster growth rate, with the highest expressing clones showing approximately a 50% reduction in doubling time. mGBP-2-expressing cells also grew to higher density and exhibited partial loss of contact growth inhibition, as evidenced by the formation of foci in post confluent cultures. In addition, mGBP-2-expressing cells showed decreased dependence on serum-derived growth factors. However, they did not lose the requirement for anchorage-dependent growth. Finally, NIH 3T3 cells expressing mGBP-2 formed tumors in athymic mice. An mGBP-2 protein carrying a point mutation (S52N) that reduced GTP binding failed to produce these phenotypes when expressed at the same levels as wild type. The additional finding that IFN-gamma treatment of NIH 3T3 cells resulted in an increase in proliferation similar to that observed for mGBP-2 in the absence of other IFN-induced proteins suggests that mGBP-2 may indeed be important for these growth changes. PMID- 11726670 TI - Versican interacts with fibrillin-1 and links extracellular microfibrils to other connective tissue networks. AB - Fibrillin-containing microfibrils are polymeric structures that are difficult to extract from connective tissues. Proteolytic digestion of tissues has been utilized to release microfibrils for study. Few of the molecules that connect microfibrils to other elements in the matrix have been identified. In this study, electron microscopic immunolocalization of anti-versican antibodies in tissues and in extracted microfibrils demonstrated that the C-terminal region of versican is found associated with fibrillin microfibrils. Extraction of microfibrils followed by treatment of microfibrils under dissociating conditions suggested that the versican C terminus is covalently bound to microfibrils. Binding assays using recombinant fibrillin-1 polypeptides and recombinant lectican lectin domains indicated that the versican lectin domain binds to specific fibrillin-1 polypeptides. The versican lectin domain also bound to molecules comigrating with authentic fibrillin-1 monomers in an assay using cell culture medium. In assays using microfibrils, the versican lectin domain demonstrated preferential binding compared with other lecticans. Binding was calcium-dependent. The binding site for versican in microfibrils is most likely within a region of fibrillin-1 between calcium-binding epidermal growth factor-like domains 11 and 21. Human mutations in this region can result in severe forms of the Marfan syndrome ("neonatal" Marfan syndrome). The connection between versican and fibrillin microfibrils may be functionally significant, particularly in cardiovascular tissues. PMID- 11726671 TI - Validated zinc finger protein designs for all 16 GNN DNA triplet targets. AB - The Cys(2)-His(2)-type zinc finger DNA-binding proteins can be engineered to bind specifically to many different DNA sequences. A single zinc finger typically binds to a 3-4-base pair DNA subsite. One strategy for design is to identify highly specific fingers that recognize each of the 64 possible DNA triplets. We started with a subgroup of the 64 triplets, the GNN-binding fingers. The GNN binding fingers have been examined in several studies, but previous studies did not produce specific fingers for all of the 16 GNN triplets. These previous studies did not provide any information on the possible positional or context effects on the performance of these fingers. To identify the most specific design and take the possible positional effects into consideration, we did a large-scale site selection experiment on our GNN designs. From this study, we identified very specific fingers for 14 of the 16 GNN triplets, demonstrating for the first time a clear positional dependence for many of the designs. Further systematic specificity study reveals that the in vivo functionality of these zinc finger proteins in a reporter assay depends on their binding affinities to their target sequences, thus giving a better understanding of how these zinc finger proteins might function inside cells. PMID- 11726672 TI - Flt-1-mediated down-regulation of endothelial cell proliferation through pertussis toxin-sensitive G proteins, beta gamma subunits, small GTPase CDC42, and partly by Rac-1. AB - Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) promotes its function primarily by activating two receptor tyrosine kinases, Flt 1 (VEGFR-1) and KDR (VEGFR-2). Recently, it has been shown that KDR is responsible for VPF/VEGF-stimulated endothelial cell (EC) proliferation and migration, whereas Flt-1 activation down-modulates KDR-mediated EC proliferation. Although KDR-mediated EC proliferation and migration have been extensively studied, much less is known about Flt-1-mediated antiproliferation. Here, we demonstrate that Flt-1-mediated antiproliferative activity can be blocked completely by the dominant negative mutant of CDC42 (CDC42-17N) and partially by a Rac1 dominant negative mutant (Rac1-17N) but is not affected by a RhoA dominant negative mutant (RhoA-19N). Both CDC42-17N and Rac1-17N increase the intracellular Ca(2+) mobilization in response to VPF/VEGF but have no effect on KDR and MAPK phosphorylation. Using the chimeric-receptor EGLT in which the extracellular domain of epidermal growth factor receptor was fused to the transmembrane and intracellular domains of Flt-1, we also demonstrate that CDC42 and Rac1 are activated by EGLT. Previously, we showed that phosphatidylinositol 3 kinase is required for Flt-1-mediated antiproliferative activity, but phospholipase C is not required. As expected, CDC42 and Rac1 activation mediated by EGLT can be completely inhibited by PI3K inhibitors, wortmannin and LY294002, and the p85 dominant negative mutant but not by either the phospholipase C inhibitor, or an intracellular Ca(2+) chilator BAPTA/AM. Surprisingly, pertussis toxin and overexpression of the free Gbetagamma-specific sequestering minigene hbetaARK1(495) also inhibit EGLT-mediated CDC42 and Rac1 activation completely. Moreover, pertussis toxin treatment also increases the intracellular Ca(2+) mobilization and inhibits the antiproliferation activity, thus suggesting that pertussis toxin-sensitive G proteins and the Gbetagamma subunits are involved in the signaling pathway of Flt-1 that down-regulates EC proliferation. Taken together, these results further expand our understanding of Flt-1-mediated antiproliferative activity in VPF/VEGF-stimulated endothelium. PMID- 11726673 TI - Agonist-induced force enhancement: the role of isoforms and phosphorylation of the myosin-targeting subunit of myosin light chain phosphatase. AB - The magnitude of agonist-induced Ca(2+) sensitization of force is tissue dependent, but an explanation for this diversity is unknown. Ca(2+) sensitization is thought to involve a G-protein-mediated inhibition of myosin light chain phosphatase activity by phosphorylation of the myosin-targeting subunit (MYPT). The MYPT has two isoforms that differ by a central insert, which lies near this phosphorylation site. Expression of MYPT isoforms is both developmentally regulated and tissue-specific. We hypothesized that the presence or absence of the central insert determines the magnitude of agonist-induced Ca(2+) sensitization. Throughout development, the chicken aorta exclusively expresses the splice-in MYPT isoform, and guanosine 5'-O-(thiotriphosphate) (GTPgammaS) produces a significant force enhancement. Early during development, the chicken gizzard expresses the splice-in MYPT isoform, and GTPgammaS produced a Ca(2+) sensitization. In the gizzard coincident with the shift in expression from the splice-in to splice-out MYPT isoform, GTPgammaS no longer produced force enhancement. In addition, adenosine 5'-O-(thiotriphosphate) (ATPgammaS) phosphorylated only adult gizzard tissue, the only tissue that did not demonstrate a Ca(2+) sensitization. These results suggest that the relative expression of splice-in/splice-out MYPT isoforms determines the magnitude of agonist-induced force enhancement and that MYPT phosphorylation is not required for Ca(2+) sensitization. PMID- 11726674 TI - How to build a better mousetrap: introduction to the special issue on methodology and design. PMID- 11726675 TI - Collecting and managing multisource and multimethod data in studies of pediatric populations. AB - OBJECTIVE: To provide recommendations for the collection and management of multisource and multimethod data in studies of children and adolescents with pediatric conditions. METHODS: We discuss limitations of single-source and single method data collection strategies. We review strategies for collecting and managing multisource and multimethod data, including coverage of the literature on level of agreement across sources, strengths and weaknesses of various source and method aggregation strategies, and methods of examining discrepancies between sources. RESULTS: Multisource and multimethod data collection strategies enable researchers to rule out alternative explanations for their findings and pose research questions that would probably not be testable with single-source, single method data sets. CONCLUSIONS: We emphasize the utility of multisource and multimethod data and provide recommendations for future work. PMID- 11726676 TI - Reporting of demographics, methodology, and ethical procedures in journals in pediatric and child psychology. AB - OBJECTIVE: To identify potential problems in methodology reporting that may limit research interpretations and generalization. METHODS: We examined the rates at which articles in four major journals publishing research in pediatric, clinical child, and child psychology report 18 important demographic, methodological, and ethical information variables, such as participants' gender, socioeconomic status, ethnicity, inclusion/exclusion criteria, and consent and assent procedures. RESULTS: Overall, participants' ages, genders, and ethnicity were reported at moderate to high rates, whereas socioeconomic status was reported less often. Reports of research methodology frequently did not include information on how and where participants were recruited, the participation/consent rates, or attrition rates. Consent and assent procedures were not frequently described. CONCLUSIONS: There is wide variability in articles reporting key demographic, methodological, and ethical procedure information. Necessary information about characteristics of participation samples, important for drawing conclusions, is lacking in the flagship journals serving the child psychology field. PMID- 11726677 TI - Developmental differences in children's use of rating scales. AB - OBJECTIVE: To examine the effect of child age and number of response choices on children's tendency to respond at the extremes of Likert-type scales rating emotional states. METHODS: Sixty children (5-6 years, 7-9 years, 10-12 years) were randomly assigned to use either three or five response choices in providing ratings in three different task conditions. Tasks were designed to have correct choices at the midpoints of the rating scales. Children also completed a self report feelings questionnaire. RESULTS: Results showed that younger children responded in an extreme manner when rating emotion-based, but not physical, tasks. Children's extreme scores did not vary as a function of number of response choices used. More extreme scores on the three tasks were related to more extreme scores on the feelings questionnaire. CONCLUSIONS: These results indicate that young children may respond in an extreme manner when rating emotional states. Researchers and clinicians should take this into account when interpreting children's self-reporting ratings. PMID- 11726678 TI - Methodological issues in outcome studies of at-risk infants. AB - OBJECTIVE: To identify methodologic problems found in follow-up studies of infants at biologic and environmental risk and provide solutions and recommendations. METHODS: This article is a literature review. RESULTS: Problems fall into four groupings: (1) conceptualization/design issues, 2) subject population concerns, 3) procedural issues, and 4) measurement/outcome concerns. CONCLUSIONS: Main-effect models are not useful; confounding and mediating variables must be identified. In addition, the following are needed: alternative analytic techniques, more precise subject selection and characterization of risk factors, geographically defined samples, broadened scope of outcome measures, and use of epidemiologic techniques. PMID- 11726679 TI - The use of focus group interviews in pediatric health care research. AB - OBJECTIVE: To review and synethize the research material on focus groups with children and adolescents and to provide guidelines for future development. METHODS: Psychlit, Medline, and Cinahl electronic databases, as well as the reference lists of those articles consulted, were reviewed for information regarding focus groups with participants under the age of 18 years. Both empirical and methodological articles were part of this review. RESULTS: We review the utility of focus groups for exploratory research, program evaluation, program development, and questionnaire construction or adaptation. Based on previous research, we provide guidelines for focus groups with children and adolescents and outline suggestions for future development. CONCLUSIONS: There is evidence to suggest that focus groups are a valuable means of eliciting children's views on health-related matters, given an appropriate research question. However, empirical research is required in order to investigate systematically the effect of different processes and variables on the final outcome of focus group interviews. PMID- 11726680 TI - Contrasts and correlations in theory assessment. AB - OBJECTIVE: To describe a systematic quantitative approach to assessing the predictions made by competing theories using contrasts and correlational indices of effect sizes. METHODS: We illustrate the use of the contrast F and t to compare and combine predictions when the raw data are continuous scores, and z contrasts when working with frequencies in 2 x k tables of counts. RESULTS: The traditional effect size correlation indicates the magnitude of the effect on individual scores of participants' assignment to particular conditions. The contrast correlation obtained from the contrast F or t is, in some cases, the easiest way of estimating the effect size correlation in designs using more than two groups. The alerting correlation is another way of appraising the predictive power of a contrast and can be used to compute the contrast F from published results when all we have are condition means and the omnibus F from an overall analysis of variance. Omnibus Fs, those with more than 1 df in the numerator, are rarely useful in data analytic work since they address unfocused questions, yielding only vague answers. CONCLUSIONS: Asking focused questions using contrasts increases the clarity of our questions and the clarity and statistical power of our answers. PMID- 11726681 TI - Daily reports and pooled time series analysis: pediatric psychology applications. AB - OBJECTIVE: To apply daily reports and pooled time series analysis (PTSA) to issues in pediatric psychology research. We discuss specific applications for this procedure in analyzing repeated observations for a small sample, including medication effects, caregiving role strain, pain reports, and treatment effects. METHODS: In the PTSA example presented, 20 daily behavior reports were provided by parents of 10 children with steroid-sensitive nephrotic syndrome (SSNS) during high-dose steroid administration and tapering. RESULTS: The full model, including child age, medication dosage, and between-subjects effects, significantly predicted children's aggressive behavior and anxious/depressed behavior. Steroid dosage significantly predicted aggressive, but not anxious/depressed, behavior. CONCLUSIONS: Daily reports analyzed using PTSA provided insight into serious behavioral side effects of steroid medications used to treat SSNS. We discuss the role of pediatric psychologists in addressing medication side effects and other time-related effects detectable using this methodology. PMID- 11726682 TI - Disentangling the effects of current age, onset age, and disease duration: parent and child attitudes toward diabetes as an exemplar. AB - OBJECTIVE: To develop a methodology for use with cross-sectional data to disentangle the effects of current age, disease onset age, and disease duration in chronically ill children. METHODS: We used a questionnaire data set from a large cross-sectional sample of mothers and children with Type 1 diabetes. The interdependence of current age, onset age, and disease duration precluded use of all three in the same regression model. Consequently, pairs of models were run, looking for a pattern in the results. RESULTS: The approach successfully disentangled the differential effects of the child's current age, disease onset age, and disease duration. Child current age predicted child attitudes about diabetes management rules, child sick-role identification, and maternal attitudes toward medical staff. Onset age predicted child-perceived family disruption and mothers' confidence in detecting a hypoglycemic reaction. Disease duration predicted maternal religious beliefs about diabetes and maternal attitudes toward medical staff. CONCLUSIONS: This study illustrates a methodology for disentangling the effects of child current age, disease onset age, and disease duration in cross-sectional data that may be useful for any childhood chronically ill population that varies in child onset age. PMID- 11726683 TI - Post-hoc probing of significant moderational and mediational effects in studies of pediatric populations. AB - OBJECTIVE: To provide examples of post-hoc probing of significant moderator and mediator effects in research on children with pediatric conditions. METHODS: To demonstrate post-hoc probing of moderational effects, significant two-way interaction effects (dichotomous variable x continuous variable; continuous variable x continuous variable) were probed with regressions that included conditional moderator variables. Regression lines were plotted based on the resulting regression equations that included simple slopes and y-intercepts. To demonstrate probing of mediational effects, the significance of the indirect effect was tested (i.e., the drop in the total predictor --> outcome effect when the mediator is included in the model), using Sobel's (1988) equation for computing the standard error of the indirect effect. RESULTS: All significant moderator and mediator effects are presented in figure form. CONCLUSIONS: The computational examples demonstrate the importance of conducting post-hoc probes of moderational and mediational effects. PMID- 11726684 TI - Methodological and statistical considerations for threats to internal validity in pediatric outcome data: response shift in self-report outcomes. AB - OBJECTIVE: To examine conceptual models of response shift, research design, and internal validity issues in the context of longitudinal outcome research using self-report measures such as pediatric quality of life. METHODS: Growth modeling was introduced and illustrated using a previously published data set (Clay, Wood, Frank, Hagglund, & Johnson, 1995) of adjustment in children with juvenile rheumatoid arthritis, diabetes, and healthy controls. RESULTS: Demonstrations revealed that growth modeling may detect response shift and may also model the time and shape of the response shift. CONCLUSIONS: Growth modeling provides one avenue to investigate response shift, thereby addressing an important threat to internal validity in longitudinal outcome research such as quality of life in children with chronic illness. PMID- 11726686 TI - Misreading of termination codons in eukaryotes by natural nonsense suppressor tRNAs. AB - Translational stop codon readthrough provides a regulatory mechanism of gene expression that is extensively utilised by positive-sense ssRNA viruses. The misreading of termination codons is achieved by a variety of naturally occurring suppressor tRNAs whose structure and function is the subject of this survey. All of the nonsense suppressors characterised to date (with the exception of selenocysteine tRNA) are normal cellular tRNAs that are primarily needed for reading their cognate sense codons. As a consequence, recognition of stop codons by natural suppressor tRNAs necessitates unconventional base pairings in anticodon-codon interactions. A number of intrinsic features of the suppressor tRNA contributes to the ability to read non-cognate codons. Apart from anticodon codon affinity, the extent of base modifications within or 3' of the anticodon may up- or down-regulate the efficiency of suppression. In order to out-compete the polypeptide chain release factor an absolute prerequisite for the action of natural suppressor tRNAs is a suitable nucleotide context, preferentially at the 3' side of the suppressed stop codon. Three major types of viral readthrough sites, based on similar sequences neighbouring the leaky stop codon, can be defined. It is discussed that not only RNA viruses, but also the eukaryotic host organism might gain some profit from cellular suppressor tRNAs. PMID- 11726688 TI - DNA sequences from multiple amplifications reveal artifacts induced by cytosine deamination in ancient DNA. AB - We show that DNA molecules amplified by PCR from DNA extracted from animal bones and teeth that vary in age between 25 000 and over 50 000 years carry C-->T and G ->A substitutions. These substitutions can reach high proportions among the molecules amplified and are due to the occurrence of modified deoxycytidine residues in the template DNA. If the template DNA is treated with uracil N glycosylase, these substitutions are dramatically reduced. They are thus likely to result from deamination of deoxycytidine residues. In addition, 'jumping PCR', i.e. the occurrence of template switching during PCR, may contribute to these substitutions. When DNA sequences are amplified from ancient DNA extracts where few template molecules initiate the PCR, precautions such as DNA sequence determination of multiple clones derived from more than one independent amplification are necessary in order to reduce the risk of determination of incorrect DNA sequences. When such precautionary measures are taken, errors induced by damage to the DNA template are unlikely to be more frequent than approximately 0.1% even under the unlikely scenario where each amplification starts from a single template molecule. PMID- 11726687 TI - The influence of DNA double-strand break structure on end-joining in human cells. AB - DNA end-joining is the major repair pathway for double-strand breaks (DSBs) in higher eukaryotes. To understand how DSB structure affects the end-joining process in human cells, we have examined the in vivo repair of linearized plasmids containing complementary as well as several different configurations of non-complementary DNA ends. Our results demonstrate that, while complementary and blunt termini display comparable levels of error-free rejoining, end-joining fidelity is decreased to varying extents among mismatched non-complementary ends. End structure also influences the kinetics of repair, accurately recircularized substrates for blunt and complementary termini being detected significantly earlier than for mismatched non-complementary ends. These results suggest that the end-joining process is composed of an early component, capable of efficiently repairing substrates requiring a single ligation event, and a late component, involved in the rejoining of complex substrates requiring multiple processing steps. Finally, these two types of repair events may have different genetic requirements as suggested by the finding that exposure of cells to wortmannin, a potent inhibitor of phosphatidylinositol 3-related kinases (PI 3-related kinases), blocks the repair of complex substrates while having little or no effect on those requiring a simple ligation event. PMID- 11726689 TI - A mRNA-based thermosensor controls expression of rhizobial heat shock genes. AB - Expression of several heat shock operons, mainly coding for small heat shock proteins, is under the control of ROSE (repression of heat shock gene expression) in various rhizobial species. This negatively cis-acting element confers temperature control by preventing expression at physiological temperatures. We provide evidence that ROSE-mediated regulation occurs at the post-transcriptional level. A detailed mutational analysis of ROSE(1)-hspA translationally fused to lacZ revealed that its highly conserved 3'-half is required for repression at normal temperatures (30 degrees C). The mRNA in this region is predicted to form an extended secondary structure that looks very similar in all 15 known ROSE elements. Nucleotides involved in base pairing are strongly conserved, whereas nucleotides in loop regions are more divergent. Base substitutions leading to derepression of the lacZ fusion at 30 degrees C exclusively resided in potential stem structures. Optimised base pairing by elimination of a bulged residue and by introduction of complementary nucleotides in internal loops resulted in ROSE elements that were tightly repressed not only at normal but also at heat shock temperatures. We propose a model in which the temperature-regulated secondary structure of ROSE mRNA influences heat shock gene expression by controlling ribosome access to the ribosome-binding site. PMID- 11726691 TI - Quantitation of telomerase components and hTERT mRNA splicing patterns in immortal human cells. AB - Telomerase is a reverse transcriptase that adds telomeric repeats to chromosomal ends. In most normal human somatic cells, telomerase is repressed and telomeres progressively shorten, leading to limited proliferative life-span. Telomerase reactivation is associated with cellular immortalization and is a frequent event during tumorigenesis. The telomerase ribonucleoprotein complex consists of two essential components, a catalytic protein subunit [human telomerase reverse transcriptase (hTERT)] and a template RNA (hTR). hTR is constitutively expressed, while hTERT is almost universally absent in telomerase-negative cells. Although repression of telomerase is transcriptional in telomerase-negative cells, post transcriptional and assembly processes are likely to play important roles in regulating telomerase activity in those that are telomerase-positive. The telomerase transcript can also be alternatively spliced into a variety of non functional forms. To establish the quantitative relationships between telomerase activity and its various components, we determined the numbers of molecules of hTR and hTERT mRNA, and the levels of alternatively spliced hTERT mRNA variants in normal, in vitro immortalized and cancer cell lines. We report here that there is surprisingly little variation in the proportion of alternatively spliced forms of hTERT in different cell lines. The only variation observed occurred when a change in splicing to non-functional forms appeared in response to conditions that repress telomerase activity in IDH4 cells. We also found that most telomerase-positive cell lines only contain a few molecules of potentially functional hTERT mRNA, and there is a correlation between telomerase activity and the levels of both hTR and hTERT +alpha+beta mRNA. PMID- 11726690 TI - Replication protein A is sequentially phosphorylated during meiosis. AB - Phosphorylation of the cellular single-stranded DNA-binding protein, replication protein A (RPA), occurs during normal mitotic cell cycle progression and also in response to genotoxic stress. In budding yeast, these reactions require the ATM homolog Mec1, a central regulator of the DNA replication and DNA damage checkpoint responses. We now demonstrate that the middle subunit of yeast RPA (Rfa2) becomes phosphorylated in two discrete steps during meiosis. Primary Rfa2 phosphorylation occurs early in meiotic progression and is independent of DNA replication, recombination and Mec1. In contrast, secondary Rfa2 phosphorylation is activated upon initiation of recombination and requires Mec1. While the primary Rfa2 phosphoisomer is detectable throughout most of meiosis, the secondary Rfa2 phosphoisomer is only transiently generated and begins to disappear soon after recombination is complete. Extensive secondary Rfa2 phosphorylation is observed in a recombination mutant defective for the pachytene checkpoint, indicating that Mec1-dependent Rfa2 phosphorylation does not function to maintain meiotic delay in response to DNA double-strand breaks. Our results suggest that Mec1-dependent RPA phosphorylation could be involved in regulating recombination rather than cell cycle or meiotic progression. PMID- 11726692 TI - Restriction enzymes increase efficiencies of illegitimate DNA integration but decrease homologous integration in mammalian cells. AB - Mammalian cells repair DNA double-strand breaks by illegitimate end-joining or by homologous recombination. We investigated the effects of restriction enzymes on illegitimate and homologous DNA integration in mammalian cells. A plasmid containing the neo(R) expression cassette, which confers G418 resistance, was used to select for illegitimate integration events in CHO wild-type and xrcc5 mutant cells. Co-transfection with the restriction enzymes BamHI, BglII, EcoRI and KpnI increased the efficiency of linearized plasmid integration up to 5-fold in CHO cells. In contrast, the restriction enzymes did not increase the integration efficiency in xrcc5 mutant cells. Effects of restriction enzymes on illegitimate and homologous integration were also studied in mouse embryonic stem (ES) cells using a plasmid containing the neo(R) gene flanked by exon 3 of HPRT: The enzymes BamHI, BglII and EcoRI increased the illegitimate integration efficiency of transforming DNA several-fold, similar to the results for CHO cells. However, all three enzymes decreased the absolute frequency of homologous integration approximately 2-fold, and the percentage of homologous integration decreased >10-fold. This suggests that random DNA breaks attract illegitimate recombination (IR) events that compete with homology search. PMID- 11726693 TI - Nearest neighbour analysis of MCM protein complexes in Drosophila melanogaster. AB - The MCM proteins are a group of six proteins whose action is vital for DNA replication in eukaryotes. It has been suggested that they constitute the replicative helicase, with a subset of the proteins forming the catalytic helicase (MCM4,6,7) while the others have a loading or control function. In this paper we show that all six MCM proteins are present in equivalent amounts in soluble extracts and on chromatin. We have also analysed soluble and chromatin associated MCM protein complexes under different conditions. This suggests that all six MCM proteins are always found in a complex with each other, although the interaction between the individual MCM proteins is not equivalent as stringent salt conditions are able to break the intact complex into a number of stable subcomplexes. These data contribute to the ongoing debate about the nature of MCM complexes, supporting the hypothesis that they act as a heterohexamer rather than as a number of different subcomplexes. Finally, using protein-protein cross linking we have shown that MCM2 interacts directly with MCM5 and MCM6; MCM5 with MCM3 and MCM2; and MCM6 with MCM2 and MCM4. This provides the first direct information about specific subunit contacts in the MCM complex. PMID- 11726694 TI - The role of basal and myogenic factors in the transcriptional activation of utrophin promoter A: implications for therapeutic up-regulation in Duchenne muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is an X-linked recessive muscle wasting disease caused by the absence of a muscle cytoskeletal protein, dystrophin. Utrophin is the autosomal homologue of dystrophin. We previously demonstrated that overexpression of utrophin in the muscles of dystrophin-null transgenic mice completely prevented the phenotype arising from dystrophin deficiency. Two independently regulated promoters control utrophin expression and the upstream promoter (promoter A) is synaptically regulated in muscle. In this study, we have investigated basal regulation and myogenic induction of promoter A. Interactions between Ap2 and Sp1 and their cognate DNA motifs are critical for basal transcription from the minimal promoter region. During differentiation of C2C12 myoblasts in vitro, a 2-fold increase in A-utrophin mRNA level was observed. Expression of a reporter gene, whose transcription was driven by a 1.3 kb promoter A fragment, paralleled expression of the endogenous transcript. Myogenic induction mapped to a conserved upstream muscle-specific E-box, which was shown to bind myogenic regulatory factors, transactivating the promoter up to 18-fold in transient assays. This study provides a basis for further understanding the regulatory mechanisms that control utrophin expression in muscle and may facilitate the development of reagents to effect therapeutic up-regulation of utrophin in DMD. PMID- 11726695 TI - Molecular recognition of pyr mRNA by the Bacillus subtilis attenuation regulatory protein PyrR. AB - The pyrimidine nucleotide biosynthesis (pyr) operon in Bacillus subtilis is regulated by transcriptional attenuation. The PyrR protein binds in a uridine nucleotide-dependent manner to three attenuation sites at the 5'-end of pyr mRNA. PyrR binds an RNA-binding loop, allowing a terminator hairpin to form and repressing the downstream genes. The binding of PyrR to defined RNA molecules was characterized by a gel mobility shift assay. Titration indicated that PyrR binds RNA in an equimolar ratio. PyrR bound more tightly to the binding loops from the second (BL2 RNA) and third (BL3 RNA) attenuation sites than to the binding loop from the first (BL1 RNA) attenuation site. PyrR bound BL2 RNA 4-5-fold tighter in the presence of saturating UMP or UDP and 150- fold tighter with saturating UTP, suggesting that UTP is the more important co-regulator. The minimal RNA that bound tightly to PyrR was 28 nt long. Thirty-one structural variants of BL2 RNA were tested for PyrR binding affinity. Two highly conserved regions of the RNA, the terminal loop and top of the upper stem and a purine-rich internal bulge and the base pairs below it, were crucial for tight binding. Conserved elements of RNA secondary structure were also required for tight binding. PyrR protected conserved areas of the binding loop in hydroxyl radical footprinting experiments. PyrR likely recognizes conserved RNA sequences, but only if they are properly positioned in the correct secondary structure. PMID- 11726696 TI - Combinatorial interactions of two amino acids with a single base pair define target site specificity in plant dimeric homeodomain proteins. AB - Four groups of plant homeodomain proteins contain a dimerization motif closely linked to the homeodomain. We here show that two sunflower homeodomain proteins, Hahb-4 and HAHR1, which belong to the Hd-Zip I and GL2/Hd-Zip IV groups, respectively, show different binding preferences at a defined position of a pseudopalindromic DNA-binding site used as a target. HAHR1 shows a preference for the sequence 5'-CATT(A/T)AATG-3', rather than 5'-CAAT(A/T)ATTG-3', recognized by Hahb-4. To analyze the molecular basis of this behavior, we have constructed a set of mutants with exchanged residues (Phe-->Ile and Ile-->Phe) at position 47 of the homeodomain, together with chimeric proteins between HAHR1 and Hahb-4. The results obtained indicate that Phe47, but not Ile47, allows binding to 5' CATT(A/T)AATG-3'. However, the preference for this sequence is determined, in addition, by amino acids located C-terminal to residue 53 of the HAHR1 homeodomain. A double mutant of Hahb-4 (Ile47-->Phe/Ala54-->Thr) shows the same binding behavior as HAHR1, suggesting that combinatorial interactions of amino acid residues at positions 47 and 54 of the homeodomain are involved in establishing the affinity and selectivity of plant dimeric homeodomain proteins with different DNA target sequences. PMID- 11726697 TI - Triplex formation by morpholino oligodeoxyribonucleotides in the HER-2/neu promoter requires the pyrimidine motif. AB - Triplex-forming oligonucleotides (TFOs) are good candidates to be used as site specific DNA-binding agents. Two obstacles encountered with TFOs are susceptibility to nuclease activity and a requirement for magnesium for triplex formation. Morpholino oligonucleotides were shown in one study to form triplexes in the absence of magnesium. In the current study, we have compared phosphodiester and morpholino oligonucleotides targeting a homopurine homopyrimidine region in the human HER2/neu promoter. Using gel mobility shift analysis, our data demonstrate that triplex formation by phosphodiester oligonucleotides at the HER-2/neu promoter target is possible with pyrimidine parallel, purine-antiparallel and mixed sequence (GT)-antiparallel motifs. Only the pyrimidine-parallel motif morpholino TFO was capable of efficient triple helix formation, which required low pH. Triplex formation with the morpholino TFO was efficient in low or no magnesium. The pyrimidine motif TFOs with either a phosphodiester or morpholino backbone were able to form triple helices in the presence of potassium ions, but required low pH. We have rationalized the experimental observations with detailed molecular modeling studies. These data demonstrate the potential for the development of TFOs based on the morpholino backbone modification and demonstrate that the pyrimidine motif is the preferred motif for triple helix formation by morpholino oligonucleotides. PMID- 11726698 TI - Strong minor groove base conservation in sequence logos implies DNA distortion or base flipping during replication and transcription initiation. AB - The sequence logo for DNA binding sites of the bacteriophage P1 replication protein RepA shows unusually high sequence conservation ( approximately 2 bits) at a minor groove that faces RepA. However, B-form DNA can support only 1 bit of sequence conservation via contacts into the minor groove. The high conservation in RepA sites therefore implies a distorted DNA helix with direct or indirect contacts to the protein. Here I show that a high minor groove conservation signature also appears in sequence logos of sites for other replication origin binding proteins (Rts1, DnaA, P4 alpha, EBNA1, ORC) and promoter binding proteins (sigma(70), sigma(D) factors). This finding implies that DNA binding proteins generally use non-B-form DNA distortion such as base flipping to initiate replication and transcription. PMID- 11726699 TI - The P1 phage replication protein RepA contacts an otherwise inaccessible thymine N3 proton by DNA distortion or base flipping. AB - The RepA protein from bacteriophage P1 binds DNA to initiate replication. RepA covers one face of the DNA and the binding site has a completely conserved T that directly faces RepA from the minor groove at position +7. Although all four bases can be distinguished through contacts in the major groove of B-form DNA, contacts in the minor groove cannot easily distinguish between A and T bases. Therefore the 100% conservation at this position cannot be accounted for by direct contacts approaching into the minor groove of B-form DNA. RepA binding sites with modified base pairs at position +7 were used to investigate contacts with RepA. The data show that RepA contacts the N3 proton of T at position +7 and that the T=A hydrogen bonds are already broken in the DNA before RepA binds. To accommodate the N3 proton contact the T(+7 )/A(+7)((')) base pair must be distorted. One possibility is that T(+7) is flipped out of the helix. The energetics of the contact allows RepA to distinguish between all four bases, accounting for the observed high sequence conservation. After protein binding, base pair distortion or base flipping could initiate DNA melting as the second step in DNA replication. PMID- 11726701 TI - Signal amplification by rolling circle amplification on DNA microarrays. AB - While microarrays hold considerable promise in large-scale biology on account of their massively parallel analytical nature, there is a need for compatible signal amplification procedures to increase sensitivity without loss of multiplexing. Rolling circle amplification (RCA) is a molecular amplification method with the unique property of product localization. This report describes the application of RCA signal amplification for multiplexed, direct detection and quantitation of nucleic acid targets on planar glass and gel-coated microarrays. As few as 150 molecules bound to the surface of microarrays can be detected using RCA. Because of the linear kinetics of RCA, nucleic acid target molecules may be measured with a dynamic range of four orders of magnitude. Consequently, RCA is a promising technology for the direct measurement of nucleic acids on microarrays without the need for a potentially biasing preamplification step. PMID- 11726700 TI - Statistical evaluation of differential expression on cDNA nylon arrays with replicated experiments. AB - In this paper we focus on the detection of differentially expressed genes according to changes in hybridization signals using statistical tests. These tests were applied to 14 208 zebrafish cDNA clones that were immobilized on a nylon support and hybridized with radioactively labeled target mRNA from wild type and lithium-treated zebrafish embryos. The methods were evaluated with respect to 16 control clones that correspond to eight different genes which are known to be involved in dorso-ventral axis specification. Moreover, 4608 Arabidopsis thaliana clones on the same array were used to judge statistical significance of expression changes and to control the false positive rates of the test decisions. Utilizing this special array design we show that differential expression of a high proportion of cDNA clones (15/16) and the respective genes (7/8) were identified, with a false positive error of <5% using the constant control data. Furthermore, we investigated the influence of the number of repetitions of experiments on the accuracy of the procedures with experimental and simulated data. Our results suggest that the detection of differential expression with repeated hybridization experiments is an accurate and sensitive way of identifying even small expression changes (1:1.5) of a large number of genes in parallel. PMID- 11726702 TI - Single tube genotyping of sickle cell anaemia using PCR-based SNP analysis. AB - Allele-specific amplification (ASA) is a generally applicable technique for the detection of known single nucleotide polymorphisms (SNPs), deletions, insertions and other sequence variations. Conventionally, two reactions are required to determine the zygosity of DNA in a two-allele system, along with significant upstream optimisation to define the specific test conditions. Here, we combine single tube bi-directional ASA with a 'matrix-based' optimisation strategy, speeding up the whole process in a reduced reaction set. We use sickle cell anaemia as our model SNP system, a genetic disease that is currently screened using ASA methods. Discriminatory conditions were rapidly optimised enabling the unambiguous identification of DNA from homozygous sickle cell patients (HbS/S), heterozygous carriers (HbA/S) or normal DNA in a single tube. Simple downstream mathematical analyses based on product yield across the optimisation set allow an insight into the important aspects of priming competition and component interactions in this competitive PCR. This strategy can be applied to any polymorphism, defining specific conditions using a multifactorial approach. The inherent simplicity and low cost of this PCR-based method validates bi directional ASA as an effective tool in future clinical screening and pharmacogenomic research where more expensive fluorescence-based approaches may not be desirable. PMID- 11726703 TI - The phytochrome A specific signaling component PAT3 is a positive regulator of Arabidopsis photomorphogenesis. AB - Phytochrome A plays a major role in early seedling development by triggering the transition from etiolated growth to greening. Seedlings germinated under constant far-red (FR) light show a partially de-etiolated phenotype that is not seen in phyA mutants. This phytochrome A specific response was used to screen a population of T-DNA mutagenized Arabidopsis seedlings. One mutant line, pat3 (phytochrome A signal transduction3), which showed no inhibition of hypocotyl elongation under FR light conditions and no FR-induced killing response, contained a T-DNA insertion in a 609-bp ORF. The recessive mutation co-segregated with the T-DNA resistance marker and could be allelic to fhy1. A 2,248-bp genomic fragment of the PAT3 locus can complement the pat3 mutant phenotype. PAT3 transcript peaked 3 d after germination and was downregulated by light. PAT3 has no significant homology to any known protein and shows no preferential cellular localization. The protein can activate transcription in yeast when fused to the GAL4 DNA-binding domain. Our results show that PAT3 is a positive regulator of phytochrome A signal transduction. PMID- 11726704 TI - Identification of a component that induces flowering of Lemna among the reaction products of alpha-ketol linolenic acid (FIF) and norepinephrine. AB - A stress-induced fatty acid [FIF; 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid] incubated with (-)-norepinephrine (NE) strongly induces flower formation in Lemna paucicostata [Yokoyama et al. (2000), Plant Cell Physiol. 41: 110). The increase of flower-inducing activity was well correlated with the decrease in FIF in the incubation mixture, and the reaction proceeded rapidly at higher pH. We detected small amounts of many active components in the mixture after incubation by HPLC analysis. In this study, two major components, named FN1 and FN2, of the reaction mixture were isolated, and their absolute stereostructures were determined. FN1 showed a strong flower-inducing activity and was identified as a tricyclic alpha-ketol fatty acid, 9(R)-11-[(2'R,8'R,10'S,11'S)-2',8'-dihydroxy-7' oxo-11'-[(Z)-2-pentenyl]-9'-oxa-4'-azatricyclo[6.3.1.0(1.5)]dodec- 5'en-10'-yl]-9 hydroxy-10-oxoundecanoic acid [corrected]. FN2, the C-9 epimer of FN1, showed no flower-inducing activity. The absolute stereostructure of FIF was also determined by a modification of Mosher's method. The 9-hydroxyl group was found to be predominantly 9R, with an enantiomeric excess of 40% (70% 9R and 30% 9S). FN1 was derived from 9R-type FIF and FN2 from 9S-type FIF. Various catecholamines and related substances were investigated for the ability to develop flower-inducing activity upon incubation with FIF. The essential structures were catechol and ethylamine groups (dopamine). PMID- 11726705 TI - Primary phloem-specific expression of a Zinnia elegans homeobox gene. AB - Some plant homeobox genes are expressed specifically in vascular cells and are assumed to function in the differentiation of specific types of vascular cells. However, homeobox genes exhibiting primary phloem-specific expression have not been reported. To elucidate the molecular mechanisms of vascular development, we undertook to isolate from Zinnia elegans primary phloem-specific homeobox genes that may function in phloem development. An HD-Zip type homeobox gene, ZeHB3, was isolated. This gene encodes a class I HD-Zip protein, and constitutes a gene subfamily with the Daucus carota gene CHB6, and Arabidopsis thaliana genes Athb 5, Athb-6, and Athb-16. In situ hybridization of 1-, 14- and 50-day-old plants demonstrated that ZeHB3 mRNA accumulation is restricted to a few cells destined to differentiate into phloem cells and to the immature phloem cells surrounding the sieve elements and companion cells. ZeHB3 protein was also localized to immature phloem cells. These findings clearly indicate that ZeHB3 is a novel homeobox gene that marks, and may function in, the early stages of phloem differentiation. PMID- 11726706 TI - Auxin- and abscisic acid-dependent osmoregulation in protoplasts of Phaseolus vulgaris pulvini. AB - Protoplasts isolated from the laminar pulvinus of Phaseolus vulgaris and bathed in a medium containing KCl as the major salt were found to swell in response to IAA and to shrink in response to ABA. The protoplasts of flexor cells and those of extensor cells responded similarly. The results indicate that the cellular content of osmotic solutes is enhanced by IAA and reduced by ABA. The IAA-induced swelling was abolished when either the K(+) or the Cl(-) of the bathing medium was replaced by an impermeant ion or when the medium was adjusted to neutral pH (instead of pH 6). The response was inhibited by vanadate. It is concluded that the swelling is caused by enhanced influxes of K(+) and Cl(-), which probably occur through K(+) channels and Cl(-)/H(+) symporters, respectively. The ABA induced shrinking was inhibited by 5-nitro-2-(3-phenylpropylamino)-benzoic acid, an anion-channel inhibitor, suggesting that it is caused by Cl(-) efflux through anion channels and charge-balancing K(+) efflux through outward-rectifying K(+) channels. It appears that the two plant hormones act on pulvinar motor cells to regulate their turgor pressure, as they do in stomatal guard cells. The findings are discussed in relation to the pulvinar movements induced by environmental stimuli. PMID- 11726707 TI - A Ca(2+)-dependent protein kinase that endows rice plants with cold- and salt stress tolerance functions in vascular bundles. AB - A rice Ca(2+)-dependent protein kinase, OsCDPK7, is a positive regulator commonly involved in the tolerance to cold and salt/drought. We carried out in situ detection of the transcript and immunolocalization of the protein. In the wild type rice plants under both stress conditions, OsCDPK7 was expressed predominantly in vascular tissues of crowns and roots, vascular bundles and central cylinder, respectively, where water stress occurs most severely. This enzyme was also expressed in the peripheral cylinder of crown vascular bundles and root sclerenchyma. Similar localization patterns with stronger signals were observed in stress-tolerant OsCDPK7 over-expressing transformants with the cauliflower mosaic virus 35S promoter. The transcript of a putative target gene of the OsCDPK7 signaling pathway, rab16A, was also detected essentially in the same tissues upon salt stress, suggesting that the OsCDPK7 pathway operates predominantly in these regions. We propose that the use of the 35S promoter fortuitously strengthened the localized expression of OsCDPK7, resulting in enhancement of the stress signaling in the inherently operating regions leading to improved stress tolerance. PMID- 11726708 TI - Relationship between the self-incompatibility and cAMP level in Lilium longiflorum. AB - The elongation of pollen tubes in Lilium longiflorum cv. Hinomoto after self incompatible pollination stopped halfway, but that after cross-compatible pollination (cross with cv. Georgia) did not. The elongation of pollen tubes after self-pollination was enhanced by exogenous cAMP and by pertussis toxin or cholera toxin, which activates adenylate cyclase. The level of endogenous cAMP in pistils after self-pollination was approximately one half of that after cross pollination. Furthermore, the activity of adenylate cyclase in pistils after self pollination was also approximately one half of that after cross-pollination. By contrast, cAMP phosphodiesterase in pistils after self-pollination was approximately 2 times as high as that after cross-pollination. A possible correlation between self-incompatibility and the low level of endogenous cAMP in lily pistils is discussed on the basis of these results. PMID- 11726709 TI - Chromatic variation of the abundance of PSII complexes observed with the red alga Prophyridium cruentum. AB - Chromatic regulation of photosystem stoichiometry in cyanophytes, green algae and probably vascular plants is achieved by regulation of the abundance of PSI in response to thylakoid electron transport state at least under our experimental conditions [cf. Fujita (1997) Photosyn. Res. 53: 83]. However, variation of not only PSI but also PSII, in reverse of each other, is characteristic of the stoichiometry regulation in red algae and some of marine cyanophytes. Our previous study with the red alga Porphyridium cruentum has revealed that PSII is inactivated by 50% upon a light shift from the light absorbed by Chl a, PSI light, to that mainly absorbed by phycobilisomes (PBS), PSII light [Fujita (1999) Plant Cell Physiol. 40: 924]. To evaluate the contribution of the photoinactivation to the chromatic variation of PSII, variation of the abundance of PSI, PSII and PBS, together with the fluorescence parameter and the activity of PSII, was followed after a light shift from PSI light to PSII light. Upon a light shift to PSII light, PSII, determined as Cyt b(559) per PBS, decreased rapidly, following the photoinactivation, down to the level a half of that before the light shift, and remained constant. Since the increase in PBS was not significant during this period, a rapid decrease of PSII/PBS led us to tentatively conclude that the degradation of PSII is a main cause for variation of the abundance of PSII. Photoinactivation of PSII, and also decrease in Cyt b(559), was accelerated, but only slightly, by the addition of chloramphenicol (CAP) at a moderate concentration while CAP at the same concentration significantly suppressed the increment of PSI determined as P700. A selective effect of CAP supports the above conclusion. PMID- 11726710 TI - An in vitro system to examine the effective phospholipids and structural domain for protein targeting to seed oil bodies. AB - An in vitro system was established to examine the targeting of proteins to maturing seed oil bodies. Oleosin, the most abundant structural protein, and caleosin, a newly identified minor constituent in seed oil bodies, were translated in a reticulocyte lysate system and simultaneously incubated with artificial oil emulsions composed of triacylglycerol and phospholipid. The results suggest that oil body proteins could spontaneously target to artificial oil emulsions in a co-translational mode. Incorporation of oleosin to artificial oil emulsions extensively protected a fragment of approximately 8 kDa from proteinase K digestion. In a competition experiment, in vitro translated caleosin and oleosin preferentially target to artificial oil emulsions instead of microsomal membranes. In oil emulsions with neutral phospholipids, relatively low protein targeting efficiency was observed. The targeting efficiency was substantially elevated when negatively charged phospholipids were supplemented to oil emulsions to mimic the native phospholipid composition of oil bodies. Mutated caleosin lacking various structural domains or subdomains was examined for its in vitro targeting efficiency. The results indicate that the subdomain comprising the proline knot motif is crucial for caleosin targeting to oil bodies. A model of direct targeting of oil-body proteins to maturing oil bodies is proposed. PMID- 11726711 TI - The response of the phosphate uptake system and the organic acid exudation system to phosphate starvation in Sesbania rostrata. AB - It is well known that the P(i) uptake system via the high-affinity P(i) transporter and the organic acid exudation system via PEPC are enhanced in the roots of P(i)-starved plants. In this paper, we compared the expression of these two systems in Sesbania rostrata, a leguminous plant, on whose roots and stems it forms nodules. When S. rostrata plants were transferred to a 0 microM P(i) nutrient solution, the expression of both the high-affinity P(i) transporter and PEPC was enhanced within 2 d. The enhancement of the expression of the high affinity P(i) transporter genes and the PEPC gene coordinated with the increases in the P(i) uptake rate and the PEPC activity, respectively. This suggests that the expression of the high-affinity P(i) transporters and PEPC is regulated in part at the transcript level. Furthermore, we examined which of the environmental or the endogenous P(i) level regulates the expression of these two systems. The P(i) content in the 6-day-old plants decreased to a lower level than that in the 15-day-old plants when grown in a 30 microM P(i) solution. At that time, the expression of the high-affinity P(i) transporters and PEPC was enhanced only in the 6-day-old plants. Moreover, the P(i) content in plants forming many nodules on their stems decreased. The expression of the high-affinity P(i) transporters and PEPC was then enhanced in the nodulated plants. These facts suggest that the expression of these two systems may be regulated by the P(i) content in the plants, not by the P(i) concentration in the soil. PMID- 11726712 TI - Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings. AB - Leaves of maize (Zea mays L.) seedlings were supplied with different concentrations of abscisic acid (ABA). Its effects on the levels of superoxide radical (O(2)(-)), hydrogen peroxide (H(2)O(2)) and the content of catalytic Fe, the activities of several antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), the contents of several non-enzymatic antioxidants such as ascorbate (ASC), reduced glutathione (GSH), alpha-tocopherol (alpha-TOC) and carotenoid (CAR), and the degrees of the oxidative damage to the membrane lipids and proteins were examined. Treatment with 10 and 100 microM ABA significantly increased the levels of O(2)(-) and H(2)O(2), followed by an increase in activities of SOD, CAT, APX and GR, and the contents of ASC, GSH, alpha-TOC and CAR in a dose- and time dependent pattern in leaves of maize seedlings. An oxidative damage expressed as lipid peroxidation, protein oxidation, and plasma membrane leakage did not occur except for a slight increase with 100 microM ABA treatment for 24 h. Treatment with 1,000 microM ABA led to a more abundant generation of O(2)(-) and H(2)O(2) and a significant increase in the content of catalytic Fe, which is critical for H(2)O(2)-dependent hydroxyl radical production. The activities of these antioxidative enzymes and the contents of alpha-TOC and CAR were still maintained at a higher level, but no longer further enhanced when compared with the treatment of 100 microM ABA. The contents of ASC and GSH had no changes in leaves treated with 1,000 microM ABA. These results indicate that treatment with low concentrations of ABA (10 to 100 microM) induced an antioxidative defence response against oxidative damage, but a high concentration of ABA (1,000 microM) induced an excessive generation of AOS and led to an oxidative damage in plant cells. PMID- 11726713 TI - Characterization of an Arabidopsis cDNA encoding a subunit of serine palmitoyltransferase, the initial enzyme in sphingolipid biosynthesis. AB - Serine palmitoyltransferase (SPT; EC 2.3.1.50) catalyzes the condensation of serine with palmitoyl-CoA to form 3-ketosphinganine in the first step of de novo sphingolipid biosynthesis. In this study, we describe the cloning and functional characterization of a cDNA from Arabidopsis thaliana encoding the LCB2 subunit of SPT. The Arabidopsis LCB2 (AtLCB2) cDNA contains an open reading frame of 1,467 nucleotides, encoding 489 amino acids. The predicted polypeptide contains three transmembrane helices and a highly conserved motif involved in pyridoxal phosphate binding. Expression of this open reading frame in the Saccharomyces cerevisiae mutant strains defective in SPT activity resulted in the expression of a significant level of sphinganine, suggesting that AtLCB2 cDNA encodes SPT. Southern blot analysis and inspection of the complete Arabidopsis genome sequence database suggest that there is a second LCB2-like gene in Arabidopsis. Expression of a green fluorescent protein (GFP) fusion product in suspension-cultured tobacco BY-2 cells showed that AtLCB2 is localized to the endoplasmic reticulum. AtLCB2 cDNA may be used to study how sphingolipid synthesis is regulated in higher plants. PMID- 11726714 TI - Functional analysis of salt-inducible proline transporter of barley roots. AB - We cloned a cDNA encoding Hordeum vulgare Proline Transporter (HvProT) from salt stressed barley roots by differential display. HvProT was 2,161 bp long and had an open reading frame encoding 450 amino acids. The deduced amino acid sequence of HvProT was similar to those of proline transporter proteins of rice (65.7%), Arabidopsis (57.7%) and tomato (42.0%). Northern blot analysis showed that the transcript level of HvProT was induced in roots at 30 min after 200 mM NaCl treatment and its peak was observed at 3 h. However, the transcript level was very low in leaves and did not increase by salt stress. The expression level of Delta(1)-pyrroline-5-carboxylate synthetase (P5CS), encoding a key enzyme of proline synthesis, was induced later than HvProT by salt stress. A transport assay using a yeast with mutation in proline uptake revealed that HvProT was a transporter with high affinity for L-proline (K(m) = 25 microM). HvProT was found to be a unique transporter with high affinity for L-proline. Since its transport activity was dependent on the pH gradient, HvProT was suggested to be a H(+)/amino acid symporter. In situ hybridization analysis showed that the HvProT mRNA was strongly expressed in root cap cells under salt stress. HvProT might play an important role in the transport of proline to root tip region urgently upon salt stress. PMID- 11726715 TI - Involvement of gibberellins in expression of a cysteine proteinase (SH-EP) in cotyledons of Vigna mungo seedlings. AB - The expression of a papain-type proteinase, designated SH-EP, in cotyledons of Vigna mungo seedlings has been shown to require some factors in the embryonic axes. Gibberellin A1 (GA(1)) and GA(20) were identified by GC-MS in embryonic axes of V. mungo seedlings. The level of accumulation of SH-EP in cotyledons of V. mungo seedlings was greatly reduced by treatment of the seeds with uniconazole P, an inhibitor for GA biosynthesis. The reduced level of accumulation of SH-EP in cotyledons by uniconazole-P was recovered by exogenous application of GA(1) and GA(20) to the seedlings. PMID- 11726717 TI - Phosphorylation, CREB, and Mental Retardation. PMID- 11726718 TI - Much huffing about HIF: oxygen dependent proline hydroxylation and the regulation of hypoxia inducible factors. PMID- 11726719 TI - Neonatal jaundice: continuing concern and need for research. PMID- 11726720 TI - Just when you thought it was safe. PMID- 11726721 TI - Bilirubin and neurological dysfunction--do we need to change what we are doing? PMID- 11726722 TI - Neonatal hyperbilirubinemia and the potential risk of subtle neurological dysfunction. PMID- 11726723 TI - Melatonin: the next panacea? PMID- 11726724 TI - Antioxidants as therapy in the newborn: some words of caution. AB - Reactive oxygen and nitrogen species are considered to play a major role in the pathogenesis of a wide range of human disorders. This may be a particularly important pathogenetic mechanism in the newborn nursery. The phrase "oxygen radical disease of prematurity" has been coined to collectively describe a wide range of neonatal disorders based on the belief that premature newborns are deficient in antioxidant defenses at a time when they are subjected to acute and chronic oxidant stresses. This belief has led to a number of clinical trials of antioxidant therapies being undertaken in neonatal patients. The realization that reactive oxygen species play a critical role in neonatal illnesses has only recently been paralleled by an increased understanding of their physiologic roles. A major concern is that effective scavenging of reactive oxygen species, to attenuate their toxic effects, will also inhibit essential cellular functions such as growth in potential target organs such as lung, brain, intestine, and retina. PMID- 11726725 TI - Apolipoprotein E phenotype regulates cholesterol absorption in healthy 13-month old children--The STRIP Study. AB - High serum cholesterol concentration is one of the key risk factors in development of atherosclerosis, which may begin early in life and later progress to symptomatic coronary heart disease. In adults, apoE polymorphism strongly influences cholesterol metabolism, as subjects with apoE 3/4 or 4/4 (collectively called apoE4) phenotype absorb cholesterol effectively and thus also have higher cholesterol absorption-reflecting plant sterol concentrations in serum than subjects with other apoE phenotypes. Because of the inverse correlation of absorption and synthesis of cholesterol, concentrations of cholesterol synthesis reflecting serum cholesterol precursor sterols are lower in subjects with apoE4 than in subjects with other phenotypes. To analyze whether apoE phenotype affects cholesterol absorption and synthesis in early childhood, we measured serum plant sterol (campesterol and sitosterol) and cholesterol precursor sterol (desmosterol and lathosterol) concentrations in healthy 13-month old children using gas-liquid chromatography. The 36 study children were participants in a randomized prospective trial (the Special Turku Coronary Risk Factor Intervention Project) aimed at decreasing exposure of the children to environmental atherosclerosis risk factors. The 16 apoE4 children had 30% to 50% higher cholesterol-adjusted campesterol and sitosterol concentrations in serum than the 20 apoE 3/3 children (p = 0.002 and p = 0.02, respectively). The concentrations of cholesterol precursor sterols in serum did not differ between the two groups of children. We conclude that the young apoE4 children may absorb cholesterol and plant sterols more effectively than the children with apoE 3/3 phenotype without compensatory reduction in endogenous synthesis of cholesterol. PMID- 11726726 TI - Early increases in brain myo-inositol measured by proton magnetic resonance spectroscopy in term infants with neonatal encephalopathy. AB - Our aim was to assess brain myo-inositol/creatine plus phosphocreatine (Cr) in the first week in term infants with neonatal encephalopathy using localized short echo time proton magnetic resonance spectroscopy and to relate this to measures of brain injury, specifically lactate/Cr in the first week, basal ganglia changes on magnetic resonance imaging (MRI), and neurodevelopmental outcome at 1 y. Fourteen term infants with neonatal encephalopathy of gestational age (mean +/- SD) 39.6 +/- 1.6 wk, birth weight 3270 +/- 490 g, underwent MRI and magnetic resonance spectroscopy at 3.5 +/- 2.1 d. Five infants were entered in a pilot study of treatment with moderate whole-body hypothermia for neonatal encephalopathy; two were being cooled at the time of the scan. T(1)- and T(2) weighted transverse magnetic resonance images were graded as normal or abnormal according to the presence or absence of the normal signal intensity of the posterior limb of the internal capsule and signal intensity changes in the basal ganglia. Localized proton magnetic resonance spectroscopy data were obtained from an 8-cm(3) voxel in the basal ganglia using echo times of 40 and 270 ms, and the peak area ratios of myo-inositol/Cr and lactate/Cr were measured. Outcome was scored using Griffith's development scales and neurodevelopmental examination at 1 y. MRI and outcome were normal in six infants and abnormal in eight. myo Inositol/Cr and lactate/Cr were higher in infants with abnormal MRI and outcome (p < 0.01, p < 0.01, respectively). myo-Inositol/Cr and lactate/Cr were correlated (p < 0.01) and were both correlated to the Griffith's developmental scales (p < 0.01, p < 0.01, respectively). In conclusion, these preliminary data suggest that early increases in brain basal ganglia myo-inositol/Cr in infants with neonatal encephalopathy are associated with increased lactate/Cr, MRI changes of severe injury, and a poor neurodevelopmental outcome at 1 y. PMID- 11726727 TI - Are moderate degrees of hyperbilirubinemia in healthy term neonates really safe for the brain? AB - In 1994 the American Academy of Pediatrics recommended more liberal rules for the treatment of hyperbilirubinemia in healthy term newborns. Yet, the safety of moderate degrees of hyperbilirubinemia in healthy term newborns is debated. To evaluate the safety of moderate degrees of hyperbilirubinemia, we assessed neurologic condition of 20 healthy nonhemolytic term newborns with peak total serum bilirubin levels of 233-444 micromol/L and 20 control infants matched for sex and gestational age at birth. Neurologic condition was evaluated with techniques focusing on the presence of minor neurologic dysfunction: in the newborn period according to Prechtl, at 3 mo on the basis of the quality of general movements, and at 12 mo according to Touwen. Moderate hyperbilirubinemia turned out to be associated with a significant increase in minor neurologic dysfunction throughout the first year of life. At 12 mo a strong dose-response relationship between the degree of hyperbilirubinemia and the severity of minor neurologic dysfunction was present. Our results indicate that total serum bilirubin levels 335 micromol/L should be avoided. PMID- 11726728 TI - Neurotoxic effects of fluorinated glucocorticoid preparations on the developing mouse brain: role of preservatives. AB - Prenatal betamethasone (Celestene) therapy reduces the incidence of brain damage, whereas prenatal or neonatal dexamethasone (Soludecadron) increases the risk of brain lesions or neuromotor deficits. To determine whether this increase is ascribable to the sulfites used as preservatives in Soludecadron, we investigated the effects of 12 h of exposure to pure dexamethasone, Soludecadron, pure betamethasone, Celestene, and sulfites on in vitro and in vivo death of neurons cultured under basal conditions or with excitotoxic agents (N-methyl-D-aspartate or (S)-5-bromowillardiine) or hypoxia. Apoptotic features were quantitated using a fluorescent chromatin stain (Hoechst 33258). Neuronal viability was unaffected by pure dexamethasone, pure betamethasone, or Celestene. Soludecadron or sulfites significantly increased neuronal loss. Pure dexamethasone or pure betamethasone produced a 40-50% decrease in neuronal death induced by N-methyl-D-aspartate, (S) 5-bromowillardiine, or hypoxia, whereas Soludecadron had no effect and sulfites significantly increased the neurotoxicity of excitotoxic agents. In in vivo experiments involving terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling after several i.p. injections of fluorinated glucocorticoids, Soludecadron, but not pure dexamethasone, significantly increased the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-stained cells in neocortical layers and thalamus. These experimental findings suggest that injectable dexamethasone should be used with caution during the perinatal period. PMID- 11726729 TI - Hypocapnia and other ventilation-related risk factors for cerebral palsy in low birth weight infants. AB - Ventilatory management patterns in very low birth weight newborns, particularly iatrogenic hypocapnia, have occasionally been implicated in perinatal brain damage. However, such relationships have not been explored in large representative populations. To examine the risk of disabling cerebral palsy in mechanically ventilated very low birth weight infants in relation to hypocapnia and other ventilation-related variables, we conducted a population-based prospective cohort study of 1105 newborns with birth weights of 500-2000 g born in New Jersey from mid-1984 through 1987, among whom 777 of 902 survivors (86%) had at least one neurodevelopmental assessment at age 2 y or older. Six hundred fifty-seven of 777 assessed survivors (85%), of whom 400 had been mechanically ventilated, had blood gases obtained during the neonatal period. Hypocapnia was defined as the highest quintile of cumulative exposure to arterial PCO(2) levels <35 mm Hg during the neonatal period. Disabling cerebral palsy was diagnosed in six of 257 unventilated newborns (2.3%), 30 of 320 ventilated newborns without hypocapnia (9.4%), and 22 of 80 ventilated newborns with hypocapnia (27.5%). Two additional ventilatory risk factors for disabling cerebral palsy were found hyperoxia and prolonged duration of ventilation. In a multivariate analysis, each of the three ventilatory variables independently contributed a 2- to 3-fold increase in risk of disabling cerebral palsy. These risks were additive. Although duration of mechanical ventilation in very low birth weight newborns likely represents severity of illness, both hypocapnia and hyperoxia are largely controlled by ventilatory practice. Avoidance of arterial PCO(2) levels <35 mm Hg and arterial PO(2) levels >60 mm Hg in mechanically ventilated very low birth weight infants would seem prudent. PMID- 11726730 TI - Effect of increased lung expansion on surfactant protein mRNA levels in lambs. AB - Increased fetal lung expansion profoundly inhibits surfactant protein gene expression and stimulates cellular proliferation in the fetal lung. Our aim was to determine whether increased expansion of the lung after birth, by the application of a continuous positive airway pressure (CPAP) for 12 h, inhibits surfactant protein gene expression and stimulates cell division in lambs. Two week-old lambs were randomly divided into 2 groups (n = 5 for each), sedated, and exposed to either no CPAP (controls) or 10 cm H(2)O of CPAP during a 12-hour treatment period. After 2 h of the treatment, (3)H-thymidine was administered to each lamb (iv) to measure pulmonary DNA synthesis rates over the following 10 h of treatment. To assess the increase in lung expansion, functional residual capacity (FRC) was measured before the start of the treatment period and again at 6 and 12 h. Compared with control lambs, a CPAP of 10 cm H(2)O increased FRC from 26.8 +/- 3.8 mL/kg to 62.9 +/- 19.7 mL/kg at 6 h and it remained elevated at 12 h (56.2 +/- 5.7 mL/kg). Despite this large increase in end expiratory lung volume (FRC), the mRNA levels for SP-A, SP-B, and SP-C and DNA synthesis rates in lung tissue were not altered. The results of this study indicate that, in contrast to the fetus, an increase in end expiratory lung volume of approximately 100% does not affect surfactant protein gene expression or pulmonary DNA synthesis rates in 2 week old lambs. Thus, the response of the lung to increases in lung expansion varies markedly before and after birth. PMID- 11726731 TI - Lipopolysaccharide-induced tumor necrosis factor-alpha and IL-10 production by lung macrophages from preterm and term neonates. AB - Lung injury in preterm neonates with respiratory failure has been attributed to persistent inflammation, which is likely to involve lung macrophages (LM). The study objective was to investigate LM during the first 8 d of life from preterm infants (n = 19), using term infants (n = 11) with respiratory failure as control subjects. LM percentages from mixed-cell suspensions produced from tracheobronchial lavage were calculated. A postnatal increase in the mean LM concentration was demonstrated within the preterm group (p = 0.01), which was greater in comparison to that from the term group (p < 0.01). Regression analyses were significant for direct relationships between LM concentrations and ex vivo lipopolysaccharide-induced tumor necrosis factor-alpha and IL-10 production (r = 0.93 and r = 0.63, respectively), establishing LM as the source of these cytokines. Comparative analyses demonstrated that the ability of preterm versus term LM to produce tumor necrosis factor-alpha was nearly identical; in contrast, a trend toward diminished levels of IL-10 expression in the preterm group was observed (p = 0.06). Thus, although studies have shown that LM precursors (i.e. cord blood monocytes) produce less tumor necrosis factor-alpha in preterm versus term infants, the present data strongly suggest that this relationship does not hold postnatally with respect to terminally differentiated LM in sick neonates. Overall, the data are consistent with a pro- versus antiinflammatory imbalance that may bear functional significance on the pathogenesis of chronic lung disease. PMID- 11726732 TI - Physical growth and retinopathy in preterm infants: involvement of IGF-I and GH. AB - GH and IGF-I are important for physical growth. We measured serum levels of these factors in preterm infants. The study population (n = 81) was divided into three groups according to the gestational age. We evaluated differences in serum GH and IGF-I levels among groups with regard to physical growth and development of retinopathy of prematurity. Serum GH levels in extremely preterm infants born at <28 wk of gestational age were significantly higher than levels in those born between 28 and 34 wk at 1 and 2 mo of age. In contrast, serum IGF-I levels in extremely preterm infants remained low, whereas those in the other two groups gradually increased. Evaluation of the effects of GH and IGF-I on physical growth in very low birth weight infants (<1500 g) showed that IGF-I concentrations were positively related to physical growth for several months after birth, whereas no relationship was observed between GH and physical growth. Multivariate analysis demonstrated that high GH concentration at 1 mo of age was significantly associated with development of severe retinopathy of prematurity. In conclusion, persistent low serum IGF-I levels may explain the slow physical growth during neonatal life, and exposure of high GH may cause, at least in part, severe retinopathy of prematurity in preterm infants. PMID- 11726733 TI - Onset of significant GH dependence of serum IGF-I and IGF-binding protein 3 concentrations in early life. AB - Forty-eight normal full-term Chinese babies (25 boys and 23 girls) were followed up every 2 mo in the first year and every 3 mo during the second year of life for anthropometric measurements. Blood samples were taken at birth and at 6, 10, 12, and 18 mo after birth for serum GH-binding protein, IGF-I, and IGF-binding protein 3 analysis. Onset of the childhood phase of growth in the infants was determined from the growth data plotted on Infancy-Childhood-Puberty growth charts. The serum GH-binding protein concentrations were low in cord blood but rose significantly at 6 mo, with slower rises in late infancy and early childhood. However, a significant rise in serum IGF-I and IGF-binding protein 3 levels was only observed from 10 mo of life onward. The change in IGF-I between birth and 6 mo was significantly correlated with length gain (r(2) = 0.35, p < 0.05) and body mass index gain (r(2) = 0.41, p < 0.01) during the same period. The 34 infants with onset of childhood phase of growth between 6 and 10 mo had a higher mean serum IGF-I value at 10 mo (8.8 +/- 5.8 nM versus 4.9 +/- 3.1 nM; p < 0.05) and higher length velocity between 10 and 12 mo (16.3 +/- 4.7 cm/y versus 8.8 +/- 4.3 cm/y; p < 0.001) compared with the 14 infants with a later onset after 10 mo of age. A significant correlation between a change in serum IGF-I and IGF-binding protein 3 levels was observed during the three 6-mo periods between birth and 18 mo, but a significant correlation between a change in serum GH binding protein and a change in serum IGF-I or IGF-binding protein 3 levels was only seen between 12 and 18 mo of age. The multiple regression analysis (r(2) = 0.43, p = 0.0002) revealed that the change in serum GH-binding protein and IGF-I concentrations between 6 and 12 mo of age and the age of onset of childhood phase of growth could explain 43% of the length gain between 6 and 12 mo of age in our babies. The results of our study support the hypothesis that the onset of the childhood phase of growth is associated with the onset of significant GH action on growth. PMID- 11726734 TI - Fas and Fas ligand expression in maternal blood and in umbilical cord blood in preeclampsia. AB - The Fas-Fas ligand (FasL) pathway of apoptosis is abnormally activated in diseases associated with impaired immune tolerance or chronic inflammation. Pregnancy-related hypertension is a spectrum of disease that commonly causes significant morbidity in women and in their newborn infants, is associated with generalized inflammation, and may be causally related to impaired maternal-fetal tolerance. Our recent observation of enhanced trophoblast expression of FasL in one form of pregnancy-related hypertension led us to hypothesize that this group of disorders might be associated with abnormal activation of the Fas-FasL pathway. To test this hypothesis, we prospectively quantified soluble and leukocyte-associated Fas receptor and FasL in the maternal and umbilical cord blood (CB) sera of 20 gestations complicated by preeclampsia and of 18 normal control gestations, using ELISA and flow cytometric analyses. We determined higher soluble FasL levels in paired maternal and CB sera of hypertensive gestations compared with control gestations (p < 0.01); in contrast, soluble Fas levels were similar between groups. Surface expression of FasL was lower on maternal (p < 0.01) and CB (p < 0.05) neutrophils from affected gestations, whereas surface Fas expression was lower on maternal (p < 0.02), but not CB, neutrophils and lymphocytes. We conclude that expression of Fas and FasL in sera and on leukocytes is altered in gestations complicated by preeclampsia, and speculate that activation of the Fas-FasL pathway mediates associated pathologic processes in affected women and in their neonates. PMID- 11726735 TI - Neonatal endotoxin exposure influences HPA responsivity and impairs tumor immunity in Fischer 344 rats in adulthood. AB - Recent research in rodents has demonstrated that exposure to bacterial endotoxin during the neonatal period alters the development of the hypothalamic-pituitary adrenal axis resulting in hypersecretion of corticosterone after stress-exposure in adulthood. Given the known interactions between glucocorticoids and the immune system it was hypothesized that such alterations may impact on immune outcomes. Fischer 344 rats were treated with endotoxin (50 microg/kg Salmonella enteritidis, i.p.) or the vehicle on postpartum d 1, 3, 5, and 7. In adulthood, animals were subjected to chronic stress (6 x 10 h/d restraint stress), and the effect on resistance to tumor colonization (experiment 1) and natural killer cell activity (experiment 2) was assessed. Experiment 3 assessed corticosterone responses to acute stress in adulthood after neonatal endotoxin or saline treatment. Neonatal endotoxin exposure resulted in a 2-fold increase in tumor colonization (p < 0.001) and a significant impairment in the activity of natural killer cells (p < 0.01), cells critically involved in the surveillance and eradication of tumor cells. Neonatal endotoxin exposure also resulted in a significant decrease in gain weight that persisted into adulthood (p < 0.05), and potentiation of corticosterone responses to acute stress in adulthood (p < 0.05). We conclude that neonatal endotoxin exposure produces long-term changes in the hypothalamic-pituitary-adrenal axis, and has significant long-term effects on immune function, specifically in terms of resistance to tumor colonization in adulthood. PMID- 11726736 TI - Effects of melatonin treatment in septic newborns. AB - Free radicals have been implicated in the pathogenesis of neonatal sepsis and its complications. This study was conducted to determine the changes in the clinical status and the serum levels of lipid peroxidation products [malondialdehyde (MDA) and 4-hydroxylalkenals (4-HDA)] in 10 septic newborns treated with the antioxidant melatonin given within the first 12 h after diagnosis. Ten other septic newborns in a comparable state were used as "septic" controls, while 10 healthy newborns served as normal controls. A total of 20 mg melatonin was administered orally in two doses of 10 mg each, with a 1-h interval. One blood sample was collected before melatonin administration and two additional blood samples (at 1 and 4 h) were collected after melatonin administration to assess serum levels of lipid peroxidation products. Serum MDA + 4-HDA concentrations in newborns with sepsis were significantly higher than those in healthy infants without sepsis; in contrast, in septic newborns treated with melatonin there was a significant reduction (p < 0.05) of MDA + 4-HDA to the levels in the normal controls at both 1 and 4 h (p < 0.05). Melatonin also improved the clinical outcome of the septic newborns as judged by measurement of sepsis-related serum parameters after 24 and 48 h. Three of 10 septic children who were not treated with melatonin died within 72 h after diagnosis of sepsis; none of the 10 septic newborns treated with melatonin died. To our knowledge, this is the first study where melatonin was given to human newborns. PMID- 11726737 TI - Localization and potential role of matrix metalloproteinase-1 and tissue inhibitors of metalloproteinase-1 and -2 in different phases of bronchopulmonary dysplasia. AB - Bronchopulmonary dysplasia (BPD) can evolve in prematurely born infants who require mechanical ventilation because of hyaline membrane disease (HMD). The development of BPD can be divided in an acute, a regenerative, a transitional, and a chronic phase. During these different phases, extensive remodeling of the lung parenchyma with re-epithelialization of the alveoli and formation of fibrosis occurs. Matrix metalloproteinase-1 (MMP-1) is an enzyme that is involved in re-epithelialization processes, and dysregulation of MMP-1 activity contributes to fibrosis. Localization of MMP-1 and its inhibitors, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2, were investigated in lung tissue obtained from infants who died during different phases of BPD development. In all studied cases (n = 50) type-II pneumocytes were found to be immunoreactive for MMP-1, TIMP-1, and TIMP-2. During the acute and regenerative phase of BPD, type-II pneumocytes re-epithelialize the injured alveoli. This may suggest that MMP-1 and its inhibitors, expressed by type-II pneumocytes, play a role in the re epithelialization process after acute lung injury. Although MMP-1 staining intensity remained constant in type-II pneumocytes during BPD development, TIMP-1 increased during the chronic fibrotic phase. This relative elevation of TIMP-1 compared with MMP-1 is indicative for reduced collagenolytic activity by type-II pneumocytes in chronic BPD and may contribute to fibrosis. Fibrotic foci in chronic BPD contained fibroblasts immunoreactive for MMP-1 and TIMP-1 and -2. This may indicate that decreased collagen turnover by fibroblasts contributes to fibrosis in BPD development. PMID- 11726738 TI - Gut-derived bone infection in the neonatal rat. AB - The risk of osteomyelitis is increased in the premature and critically ill neonate. Although potential sites of bacterial entry are present in many of these infants, the source of infection frequently cannot be established. This study was performed to assess the possible role of bacterial translocation from the intestine in the origin of bone infection using models of breast-fed and formula fed rat pups. Newborn Sprague-Dawley rats suckled either ad libitum by the dam (n = 30), or were fed a rat milk-simulated formula (n = 30). After 3 d, the animals were killed, and the left femur, heart blood, mesenteric lymph nodes, liver, spleen, and terminal ileum were excised. Organs were analyzed for bacteria by standard microbiologic procedures. Bacterial translocation occurred in 23% of breast-fed rats; the bone was not infected in any of these animals. After feeding of formula diet, bacterial counts of the ileum were markedly elevated (p < 0.001), and the composition of the gut flora was disrupted. Bacterial translocation was noted in all formula-fed rats. Bone cultures were positive in 23 of 30 (77%) rats after formula-feeding (p < 0.001 versus breast-feeding). Organisms translocated to the bone included Enterococci, Proteus, Enterobacter, and Escherichia coli. Bacterial species cultured from the bone correlated with the individual colonization pattern of other extraintestinal organs and with the composition of the ileal flora. Members of the gut flora can escape the intestine and colonize the bone in formula-fed rats. The gut should be considered as a potential source for osteomyelitis in the neonate. PMID- 11726739 TI - Donut shop diversion program. PMID- 11726741 TI - Datapoints: suicide and access to care. PMID- 11726742 TI - Consumer & family information: borderline personality disorder. PMID- 11726743 TI - Emergency psychiatry: tools of engagement: avoiding pitfalls in collaborating with patients. PMID- 11726744 TI - Practical geriatrics: insomnia in late life. PMID- 11726745 TI - Personal accounts: us and them. PMID- 11726747 TI - Economic grand rounds: Is there a shortage of psychiatrists? PMID- 11726748 TI - Policy implications for implementing evidence-based practices. AB - The authors describe the policy and administrative-practice implications of implementing evidence-based services, particularly in public-sector settings. They review the observations of the contributors to the evidence-based practices series published throughout 2001 in Psychiatric Services. Quality and accountability have become the watchwords of health and mental health services; evidence-based practices are a means to both ends. If the objective of accountable, high-quality services is to be achieved by implementing evidence based practices, the right incentives must be put in place, and systemic barriers must be overcome. The authors use the framework from the U.S. Surgeon General's 1999 report on mental health to describe eight courses of action for addressing the gap between science and practice: continue to build the science base; overcome stigma; improve public awareness of effective treatments; ensure the supply of mental health services and providers; ensure delivery of state-of-the art treatments; tailor treatment to age, sex, race, and culture; facilitate entry into treatment; and reduce financial barriers to treatment. PMID- 11726749 TI - Strategies for disseminating evidence-based practices to staff who treat people with serious mental illness. AB - Evidence-based practices have not been widely implemented in real-world treatment settings for several reasons, including existing state laws, administrative policies, funding priorities, advocates' concerns, and program staffing. Dissemination strategies focus largely on program staffing and the question of why treatment teams that are responsible for assisting people with serious mental illness fail to use evidence-based practices. In a review of the research literature, two barriers to staff dissemination emerge: individual service providers lack the necessary knowledge and skills to assimilate these practices, and certain organizational dynamics undermine the treatment teams' ability to implement and maintain innovative approaches. Three sets of strategies are useful for overcoming these barriers and fostering dissemination: packaging evidence based practices so that specific interventions are more accessible and user friendly to service providers; educating providers about relevant knowledge and skills; and addressing the organizational dynamics of the team to facilitate the implementation of innovations. Research on dissemination is relatively new and is less well developed than the clinical and services research enterprise that has led to evidence-based practices. Implications for future studies are discussed. PMID- 11726750 TI - Organizational process: a missing link between research and practice. AB - Organizational process is an underexamined barrier and a potential bridge for the introduction of innovative treatment models into mental health practice. The author describes key operational characteristics of large, complex organizations and strategies that have been used to facilitate implementation of innovative programs in the Department of Veterans Affairs health care system. He argues that complex organizations of the type in which mental health care is increasingly delivered are characterized by multiple competing goals, uncertain technologies, and fluid involvement of key participants. Interventions shown to be effective in controlled studies are often not easily introduced into such organizations, because research is typically conducted in a buffered organizational niche that is shielded from the complex open systems around it. Key strategies for moving research into practice include constructing decision-making coalitions, linking new initiatives to legitimated goals and values, quantitatively monitoring implementation and ongoing performance, and developing self-sustaining communities of practice as well as learning organizations. The author shows how effective dissemination of new treatment methods requires attention to and effective engagement with organizational processes. PMID- 11726751 TI - Special section on stigma as a barrier to recovery: Introduction. PMID- 11726752 TI - Stigma as a barrier to recovery: Perceived stigma and patient-rated severity of illness as predictors of antidepressant drug adherence. AB - OBJECTIVE: Major depression is undertreated despite the availability of effective treatments. Psychological barriers to treatment, such as perceived stigma and minimization of the need for care, may be important obstacles to adherence to the pharmacologic treatment of major depression. The authors examined the impact of barriers that were present at the initiation of antidepressant drug therapy on medication adherence in a mixed-age sample of outpatients with major depression. METHODS: A two-stage sampling design was used to identify adults with a diagnosis of major depressive disorder, as determined by the Structured Clinical Interview for Diagnosis, who sought mental health treatment at outpatient clinics. Additional instruments were administered to 134 newly admitted adults who had been taking a prescribed antidepressant medication for at least a week to assess perceived stigma, self-rated severity of illness, and views about treatment. The patients were reinterviewed three months later and were classified as adherent or nonadherent on the basis of self-reported estimates of the number and frequency of missed doses. RESULTS: Medication adherence was associated with lower perceived stigma, higher self-rated severity of illness, age over 60 years, and absence of personality pathology. No other characteristics of treatment or illness were significantly related to medication adherence. CONCLUSIONS: Perceived stigma associated with mental illness and individuals' views about the illness play an important role in adherence to treatment for depression. Clinicians' attention to psychological barriers early in treatment may improve medication adherence and ultimately affect the course of illness. PMID- 11726753 TI - Stigma as a barrier to recovery: The consequences of stigma for the self-esteem of people with mental illnesses. AB - OBJECTIVE: The objective of this study was to determine whether stigma affects the self-esteem of persons who have serious mental illnesses or whether stigma has few, if any, effects on self-esteem. METHODS: Self-esteem and two aspects of stigma, namely, perceptions of devaluation-discrimination and social withdrawal because of perceived rejection, were assessed among 70 members of a clubhouse program for people with mental illness at baseline and at follow-up six and 24 months later. RESULTS: The two measures of perceptions of stigma strongly predicted self-esteem at follow-up when baseline self-esteem, depressive symptoms, demographic characteristics, and diagnosis were controlled for. Participants whose scores on the measures of stigma were at the 90th percentile were seven to nine times as likely as those with scores at the 10th percentile to have low self-esteem at follow-up. CONCLUSIONS: The stigma associated with mental illness harms the self-esteem of many people who have serious mental illnesses. An important consequence of reducing stigma would be to improve the self-esteem of people who have mental illnesses. PMID- 11726754 TI - Stigma as a barrier to recovery: Adverse effects of perceived stigma on social adaptation of persons diagnosed with bipolar affective disorder. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of concerns about stigma on social adaptation among persons with a diagnosis of bipolar affective disorder. METHODS: The sample comprised 264 persons who were consecutively admitted to a psychiatric inpatient or outpatient service at a university affiliated hospital and who met research diagnostic criteria for bipolar I disorder, bipolar II disorder, or schizoaffective disorder, manic type. Patients were evaluated with use of the Schedule for Affective Disorders and Schizophrenia, Lifetime Version (SADS-L), the Brief Psychiatric Rating Scale (BPRS), and a measure of perceived stigma. Social adjustment was measured at baseline and seven months later with the Social Adjustment Scale (SAS). RESULTS: As predicted, patients who had concerns about stigma showed significantly more impairment at seven months on the social leisure subscale but not on the SAS extended family subscale, after baseline SAS score and symptom level had been controlled for. More refined models using SAS-derived factors as dependent variables indicated that concerns about stigma predicted higher avoidance of social interactions with persons outside the family and psychological isolation at seven-month follow-up, after baseline SAS and BPRS scores had been controlled for. CONCLUSIONS: Concerns about the stigma associated with mental illness reported by patients during an acute phase of bipolar illness predicted poorer social adjustment seven months later with individuals outside the patient's family. Greater attention to patients' concerns about stigma is needed from both researchers and clinicians. PMID- 11726755 TI - Stigma as a barrier to recovery: The extent to which caregivers believe most people devalue consumers and their families. AB - OBJECTIVE: The extent to which 461 caregivers of persons with serious mental disorders believed that most people devalue consumers and their families was assessed, and the magnitude of the relationships between these beliefs and the diagnostic status of consumers was estimated. METHODS: Caregivers of 180 consumers with schizophrenia, major depression, or bipolar disorder and caregivers of 281 consumers with bipolar disorder or schizoaffective disorder, manic type, completed a 15-item instrument comprising two scales: eight of the 15 items operationally defined the devaluation of individual consumers, and seven items operationally defined the devaluation of consumers' families. RESULTS: No significant differences were observed between the two samples on the two devaluation scales or on 14 of the 15 items that constituted the scales. About 70 percent of all caregivers indicated a belief that most people devalue consumers, and 43 percent expressed a belief that most people also devalue the families of consumers. CONCLUSIONS: Strong evidence from previous research indicates that the caregiving role is very demanding, is frequently distressing, and may be harmful to health and injurious to one's quality of life. The addition of a community that is perceived to be rejecting makes life even more difficult for the caregivers and families of people with serious mental disorders. The development and implementation of effective interventions to create more supportive and understanding communities would be a challenging and worthwhile endeavor. PMID- 11726756 TI - Depression in the workplace: costs and barriers to treatment. AB - Surveys estimate that 1.8 to 3.6 percent of workers in the U.S. labor force suffer from major depression. Depression has a significant impact on vocational functioning. Seventeen to 21 percent of the workforce experiences short-term disability during any given year, and 37 to 48 percent of workers with depression experience short-term disability. Studies indicate that treating workplace depression provides favorable cost offsets for employers, although a number of methodological issues have influenced the interpretation of these findings. In addition to disability costs, cost analyses need to include lost wages and indirect costs to employers, such as the costs of hiring and training new employees. In general, employers are not aware of the extent of the indirect costs of untreated depression. They have mistaken assumptions about the availability of effective treatment, and they are unaware of how often depression contributes to worker disability. The workers' compensation system and the courts have been slow to recognize depression as a work-related disability, and as a result employers have few incentives to treat and prevent workplace depression. PMID- 11726757 TI - Characteristics and service use patterns of nonelderly medicare beneficiaries with schizophrenia. AB - OBJECTIVE: The authors sought to describe the characteristics of nonelderly Medicare beneficiaries who have a diagnosis of schizophrenia and to assess the impact of Medicare financing on service quality by comparing service use among individuals who were enrolled only in Medicare and those who were enrolled in both Medicare and Medicaid. The authors hypothesized that persons who received only Medicare benefits would use proportionally fewer psychosocial services and less antipsychotic medication than individuals who were dually enrolled. METHODS: Data were drawn from the Medicare Current Beneficiary Survey (MCBS). The study sample consisted of 257 individuals younger than age 65 who were included in the 1995 MCBS sample and who had one inpatient or two outpatient claims for schizophrenia between 1992 and 1996. The variables examined were demographic characteristics, comorbid psychiatric and substance use disorders, activities of daily living, instrumental activities of daily living, self-reported use of prescribed antipsychotic medication, and claims for psychosocial services. A multivariate analysis was also conducted to predict the use of antipsychotic medication from demographic and health status variables. RESULTS: Dually enrolled beneficiaries were significantly more likely to be receiving antipsychotic medication than Medicare-only beneficiaries, even when the analysis controlled for demographic characteristics, health status, and comorbidity. No significant differences were found in the use of psychosocial services. CONCLUSIONS: The findings were consistent with the hypothesis that Medicare financing, which restricts access to many mental health services, is not conducive to good community care for persons with schizophrenia. PMID- 11726758 TI - A program for teaching psychiatric residents to provide integrated psychiatric and primary medical care. AB - The Psychiatry Primary Medical Care program of the Portland Veterans Affairs Medical Center teaches psychiatric residents to provide integrated psychiatric and medical care in a primary care setting. During the program's first year of operation, 34 patients received ongoing integrated care from seven residents. Patients, psychiatric residents, and medical faculty reported a high degree of satisfaction with the program. The duration and frequency of visits reflected the substantial mental health and primary care needs of the population. Patients frequently missed appointments. Barriers to starting and maintaining the program included space constraints and the amount of supervision required. PMID- 11726759 TI - A survival strategy for an academic psychiatry department in a managed care environment. AB - The changing economics of medical practice have had a profound effect on the educational, research, and service missions of academic departments of psychiatry across the country. The authors describe the development of a managed behavioral health care organization in their parent academic health system as a survival strategy for allowing their department to function in a managed care environment. They present a series of lessons learned in this effort to adapt to a highly volatile managed behavioral health care market: know how you fit into your market as well as your institution, form cooperative alliances within and outside of your institution, provide incentives to manage risk, focus on core competencies, innovate in your areas of strength, and collect data. PMID- 11726761 TI - Insight and disability. PMID- 11726760 TI - Encouraging pursuit of court-ordered treatment in a state hospital. AB - The authors discuss a performance improvement project designed to encourage staff psychiatrists at a large state hospital to more assertively pursue the option of court-ordered treatment for severely mentally ill inpatients who refuse medication for more than a week. A campaign was undertaken in 1997 to educate psychiatrists about the importance of using psychotropic medications early in a patient's hospital stay. A monthly report is now circulated to all personnel identifying units on which patients have refused medication for more than a week and psychiatrists who have filed court petitions. The number of petitions filed annually has doubled, from 97 in the year before the project to 192 four years later. PMID- 11726762 TI - Rates of HIV infection among probationers and parolees with mental illness. PMID- 11726763 TI - Telepsychiatry in a rural inpatient setting. PMID- 11726765 TI - Recognizing and managing "illy" intoxication. PMID- 11726766 TI - Calpain inhibitors prevent nitric oxide-triggered excitotoxic apoptosis. AB - The pathogenesis of some neurodegenerative disorders has been linked to excitotoxicity, excess generation of nitric oxide (NO) and apoptosis. Here, we used a model of NO-triggered neuronal apoptosis that was strictly dependent on autocrine NMDA receptor (NMDA-R) activation and intracellular Ca2+ increase. We investigated the efficiency and potentially beneficial effects of calpain inhibition. Three calpain inhibitors that prevented intracellular fodrin proteolysis also blocked apoptotic features such as decrease in mitochondrial membrane potential, chromatin breakdown, and subsequent death of cerebellar granule neurons exposed to NO donors (S-nitroso-L-glutathione, S-nitroso-N-acetyl d,l-penicillamine, and diethylamino-diazenolate-2-oxide). Since inhibitors did not interfere with NMDA-R activation, we suggest that block of calpains blunts NO triggered neuronal apoptosis by stopping the cascade downstream of primary autocrine excitotoxic events. PMID- 11726767 TI - Activity-dependent accumulation of Ca2+ in axon and dendrites of the leech Leydig neuron. AB - We have investigated Ca2+ changes evoked by single action potentials (APs) in axon and dendrites of leech Leydig neurons. Dendritic Ca2+ transients induced by an AP were twice as large as in the axon, and Ca2+ recovery was significantly faster in the dendrites as compared to the axon. The AP-induced Ca2+ transients were blocked by Co2+ and suppressed in Ca2+-free saline, indicating Ca2+ influx through voltage-activated channels. During a train of APs, Ca2+ accumulated significantly more in the axon than in the dendrites. Suppression of the Ca2+ influx changed the shape of the action potential and increased the firing frequency. The results suggest a functional role of Ca2+ influx and Ca2+ accumulation during electrical activity in different neuronal subcompartments. PMID- 11726768 TI - Decreased pallidal GABA following reverse microdialysis with clozapine, but not haloperidol. AB - Changes in striatopallidal GABA are believed to play a significant role in the motor side effects produced by antipsychotic drugs (APDs). In the current study, we measured extracellular GABA in the globus pallidus (GP) of rats. GABA release was partially impulse- and Ca2+-dependent, as evidenced by decreased efflux following tetrodotoxin (TTX) or removal of Ca2+. In addition, GABA release was significantly increased by high K+ (100 mM KCl) stimulation. Reverse dialysis of the atypical APD, clozapine (1-100 microM), produced a concentration dependent decrease in extracellular GABA. In contrast, the typical APD, haloperidol (1-100 microM), had no significant effect on GABA levels. These results suggest that clozapine has direct actions within the GP, while the effects of haloperidol are most likely mediated through its effects in the striatum. The clozapine-induced decrease in pallidal GABA may account for its low motor side effect liability. PMID- 11726769 TI - Effects of ATP on TTX-sensitive and TTX-resistant sodium currents in rat sensory neurons. AB - Differential effects of ATP on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin resistant (TTX-R) sodium currents in rat dorsal root ganglion neurons were studied using the whole-cell variation of path-clamp technique. Currents were evoked by step depolarizations to 0 mV from a holding potential of -80 mV. ATP suppressed TTX-S sodium currents while it increased TTX-R sodium currents. The effects were concentration-dependent and were reversible upon washout with ATP free external solution. ATP-gamma-S, a hydrolysis-resistant ATP analog, also affected two types of sodium currents similarly to ATP, excluding the possibility that the effects were caused by the products of ATP hydrolysis, namely adenosine. ATP by modulating sodium currents may exert profound effects on the transmission of sensory information such as nociception. PMID- 11726770 TI - NK1 receptor and the ventral medulla of the rat: bulbospinal and catecholaminergic neurons. AB - Bulbospinal C1 neurons are sympathoexcitatory and excited by substance P. However the substance P receptor (NK1 receptor), has been reported to be absent from the somata of C1 neurons. In this study, using double and triple labelling immunofluorescence and retrograde tracing, we provide evidence that the NK1 receptor is present on 5.3% of C1 neurons, and that 4.7% of C1 neurons receive close oppositions from NK1 receptor immunoreactive terminals, indicating a pre synaptic and post-synaptic site for the action of substance P. These results provide support for the sympathoexcitatory actions of substance P on C1 neurons. We also demonstrate the NK1 receptor on bulbospinal neurons of the ventral respiratory group, in a region overlapping the pre-Botzinger Complex. PMID- 11726771 TI - Is perceptual anticipation a motor simulation? A PET study. AB - A large body of psychophysical evidence suggests that perception of human movement is constrained by the observer's motor competence. PET measurements of regional cerebral blood flow were performed in eight healthy subjects who were requested, in a forced-choice paradigm, to anticipate the outcome of a single moving dot trajectory depicting the beginning of either mechanical, pointing, or writing movements. Selective activation of the left premotor cortex and of the right intraparietal sulcus was associated with visual anticipation of pointing movements while the left frontal operculum and superior parietal lobule were found to be activated during anticipation of writing movements. These results are discussed in the perspective that the motor system is part of a simulation network, which is used to interpret perceived actions. PMID- 11726772 TI - Presence of kynurenic acid and kynurenine aminotransferases in the inner retina. AB - Kynurenine aminotransferases (KATs I and II) are pivotal to the synthesis of kynurenic acid (KYNA), the only known endogenous glutamate receptor antagonist and neuroprotectant. This study is the first to identify KYNA in the rat retina and to examine immunohistochemically the distribution of KAT isoforms. As determined by HPLC, KYNA concentration in the retina was 99.9 +/- 24.6 pmol/g wet wt. Immunohisto- chemical experiments showed that both KATs were present in the retina. KAT I was preferentially localised on Muller cell endfeet while KAT II was expressed in cells within the ganglion cell layer. In conclusion, KYNA is present and synthesised in the inner retina. This may suggest a modulatory role in glutamate-mediated retinal neurotransmission. PMID- 11726773 TI - Transient spatial attention modulates distinct components of the auditory ERP. AB - We recorded ERPs to pairs of externally presented tones, T1 and T2, in the absence of attentional cues to determine whether attention is momentarily sustained at the location of a behaviourally relevant sound, and what effect this focusing of attention might have on the neural response to target stimuli. ERPs to T2 were more negative when the preceding T1 was presented on the same side of fixation than when T1 was presented on the opposite side of fixation. This negative difference consisted of an early, parietal phase and a later, frontocentral phase. These results confirm and extend previously reported effects of transient spatial attention on auditory ERPs, and they demonstrate that transient spatial attention has a distinct and robust effect on the early stages of stimulus processing in the auditory system. PMID- 11726774 TI - Dissociation of reward anticipation and outcome with event-related fMRI. AB - Reward processing involves both appetitive and consummatory phases. We sought to examine whether reward anticipation vs outcomes would recruit different regions of ventral forebrain circuitry using event-related fMRI. Nine healthy volunteers participated in a monetary incentive delays task in which they either responded to a cued target for monetary reward, responded to a cued target for no reward, or did not respond to a cued target during scanning. Multiple regression analyses indicated that while anticipation of reward vs non-reward activated foci in the ventral striatum, reward vs non-reward outcomes activated foci in the ventromedial frontal cortex. These findings suggest that reward anticipation and outcomes may differentially recruit distinct regions that lie along the trajectory of ascending dopamine projections. PMID- 11726775 TI - CB1-cannabinoid and mu-opioid receptor co-localization on postsynaptic target in the rat dorsal horn. AB - Cannabinoids and opioids interact in the control of nociception at the spinal level. Likely, several mechanisms are involved, with one of them being co localization of cannabinoid and opioid receptors. In order to validate this hypothesis, a double labeling study of CB1 cannabinoid receptors and mu-opioid receptors in the dorsal horn of the rat spinal cord was performed. A strong co localization of CB1 and mu-opioid receptors was observed in lamina II interneurons at the ultrastructural level. The physiological consequences of the co-localization are discussed. PMID- 11726776 TI - Vanilloid receptor homologue, VRL1, is expressed by both A- and C-fiber sensory neurons. AB - The capsaicin receptor (VR1) homologue, VRL1, is thought to be responsible for transducing high-threshold heat responses in Adelta-fiber neurons. In the present study, the expression of VRL1 by A- or C-fiber sensory neurons in rats was investigated by using a VRL1 and 200 kDa neurofilament (NF200, an A-fiber marker) double immunohistochemical staining method. Approximately 46% of VRL-positive neurons were NF200 positive. Though double-labeled neurons tended to be medium to large, many VRL1 single-labeled neurons were large. Dense VRL1 immunoreactivity was also found in laminae I and II of the spinal dorsal horn, where nociceptive Adelta- and C-fibers normally terminate. These results suggest that both C-fiber and Adelta-fiber primary sensory neurons express VRL1, and VRL1 may play an important role in nociception. PMID- 11726777 TI - The hemispheric difference of semantic processing of Chinese characters in two dimensions as revealed by ERPs. AB - An experiment was conducted to examine the hemispheric dominance in semantic processing of Chinese characters. Results showed that N1 is the earliest component that reflects the semantic processing of Chinese characters. Although N1 of the left hemisphere is larger in amplitude, that of the right hemisphere is shorter in latency. Based on these findings, the authors propose that the left hemisphere does not start processing until the necessary information has been transferred from the right hemisphere. Once the left hemisphere starts processing, its intensity is stronger. Thus, it is more appropriate to differentiate hemispheric processing dominance into two dimensions: speed and intensity. Moreover, the semantic cognition onset of Chinese characters occurs from 100 ms to 160 ms in the posterior perceptual processing area of brain. PMID- 11726778 TI - Melanocortin receptor agonist transiently increases oxygen consumption in rats. AB - Acute injections of melanocortin (MC) agonist and antagonist are highly effective in reducing or increasing food take, respectively. Much less is known about how injection of MC receptor active substances affects metabolism, in particular during long term administration. Here we investigated the effect of 8 days continuous i.c.v. infusion of either MC receptor agonist MTII or the selective MC4 receptor antagonist HS024 on oxygen consumption, food intake and body weight in rats. We observed significant increase in oxygen consumption 2 days after the start of the MTII infusion. However, this increase had disappeared by day 4 of the study. No difference was observed in the oxygen consumption after injection of HS024. MTII substantially decreased the food intake during the first days, but then the feeding recovered and the body weight stabilised at a new level. The immediate effect of the MC receptor agonist on both food intake and metabolism was thus transient, even though the weight loss was maintained. The HS024 treated rats were hyperphagic throughout the test period, continuously gaining weight, resulting in increased fat pads and high leptin levels. This is the first study that describes long term effects of MC receptor agonist and antagonist on metabolism and energy balance. PMID- 11726779 TI - Effects of cerebellar dentate nucleus GABAergic cells on rat inferior olivary neurons. AB - The present study was undertaken to analyze the effects on unitary activity of inferior olive (IO) neurons elicited by activation of cerebellar lateral nucleus (LN), in rats submitted to the chronic destruction of MDJ structures, i.e. in animals in which the LN-evoked effects in IO should be depended only on activation of GABAergic cells of LN. It has been observed that about two-thirds of the olivocerebellar neurons are significantly affected by LN stimulation, and > 68% of those cells were inhibited. Two-thirds of the inhibitory responses were compatible with a monosynaptic linkage, whereas the remaining inhibitions were probably due to polisynaptic linkages. The majority of LN-induced inhibitions was abolished or greatly reduced following application of GABA antagonists. PMID- 11726780 TI - The T. cruzi trans-sialidase induces PC12 cell differentiation via MAPK/ERK pathway. AB - The trans-sialidase (TS) of Trypanosoma cruzi induces survival and differentiation of neuronal and glial cells. This mechanism underlying survival is via phosphatidylinositol 3-kinase (PI3K) but how TS promotes neuronal differentiation remained to be determined. Here we show that TS-induced neurite outgrowth in PC12 cells is through sustained activation of the mitogen-activated protein kinase or ERK cascade and, therefore, by a signaling mechanism distinct from the one it uses to induce cell survival. Such differential activation of signal pathways in neuronal cells to effectuate diverse biological activities is analogous to the action of authentic neurotrophins and other growth factors, thereby reinforcing the novel concept of T. cruzi mimicry of host neurotrophic factor(s). PMID- 11726781 TI - The direct effect of insulin on barosensitive neurones in the nucleus tractus solitarii of rats. AB - The present investigation was designed to determine the direct effect of insulin on the spontaneous discharge of barosensitive neurones in the nucleus tractus solitarii (NTS) of rats anaesthetized with urethane. Microinjection of 20 nl insulin (10 IU/ml) into NTS decreased the spontaneous discharge of 38 of the 52 units studied (73.1%), and this decrease was augmented by increasing the concentration to 40 IU/ml. Microinjections of insulin vehicle, glucose, hydralazine or phenylephrine did not elicit significant changes in the spontaneous discharge of NTS barosensitive neurones. These results demonstrate that insulin inhibits the spontaneous discharge of barosensitive NTS neurones. They suggest that insulin increases sympathetic nervous activity via a central neural mechanism and may play a role in the modulation of cardiovascular information within the NTS. PMID- 11726782 TI - Endomorphin-1 improves scopolamine-induced impairment of short-term memory via mu1-opioid receptor in mice. AB - The effects of intracerebroventricular injection of endomorphin-1 and 2, endogenous mu-opioid receptor agonists, on the scopolamine-induced impairment of spontaneous alternation performance associated with short-term memory were investigated in mice. Endomorphin-1 (0.03 microg) inhibited scopolamine (1 mg/kg) induced impairment of spontaneous alternation performance without affecting total arm entries, while endomorphin-2 (0.01-10 microg) failed to significantly influence the scopolamine (1 mg/kg)-induced impairment. Endomorphin-1 (0.03 microg) itself had no marked effects on spontaneous alternation performance in intact mice. Although beta-funaltrexamine (5 microg), a mu-opioid receptor antagonist, did not significantly affect the inhibitory effects of endomorphin-1 (0.03 microg) on the scopolamine (1 mg/kg)-induced impairment, naloxonazine (35 mg/kg), a mu1-opioid receptor antagonist, significantly reversed the inhibitory effects of endomorphin-1 (0.03 microg) on the impairment. Naloxonazine (35 mg/kg) unlike beta-funaltrexamine (5 microg) did not significantly influence the scopolamine (1 mg/kg)-induced impairment of spontaneous alternation performance. These results suggest that endomorphin-1 improves the disturbance of short-term memory resulting from cholinergic dysfunction through the mediation of mu1-opioid receptors. PMID- 11726783 TI - Estimation of aspartate synthesis in GABAergic neurons in mice by 13 C NMR spectroscopy. AB - Aspartate synthesis in GABAergic neurons was estimated following inhibition of glutamate decarboxylase (GAD) with 3-mercaptopropionic acid (3-MPA). Mice received 3-MPA, 50 mg/kg, and [1-13C]glucose or [2-13C]acetate. Brain extracts were analyzed by 13C NMR spectroscopy. GABA synthesis was inhibited by 50%, and the synthesis of [13C]aspartate subsequently decreased by 25%. This means that 50% of cerebral aspartate is labeled from metabolites formed through the GABA shunt. A large proportion of the remaining aspartate is labeled through the TCA cycle in GABAergic neurons. PMID- 11726784 TI - Cloning and mutation analysis of the human potassium channel KCNQ2 gene promoter. AB - Benign familial neonatal convulsions (BFNC) have been previously found to be associated with mutations within the coding region of KCNQ2. We have now cloned and analyzed the promoter region of the human KCNQ2 gene. 5'-RACE identified a transcription start site (TSS) located 200 bp upstream of the ATG start codon. The TSS is located close to a repetitive region containing seven copies of a degenerate 42-mer repeat. Several different luciferase (LUC) reporter plas- mids containing fragments from the KCNQ2 5'-flanking region were constructed and expressed in NT2N and SH-SY5Y cell lines. A core promoter region was found to be located between bp 20 and bp 74 upstream of the TSS. Neither the promoter region nor the repetitive region showed any mutations in 13 index patients from unrelated BFNC families. PMID- 11726785 TI - How the brain perceives causality: an event-related fMRI study. AB - Detection of the causal relationships between events is fundamental for understanding the world around us. We report an event-related fMRI study designed to investigate how the human brain processes the perception of mechanical causality. Subjects were presented with mechanically causal events (in which a ball collides with and causes movement of another ball) and non-causal events (in which no contact is made between the balls). There was a significantly higher level of activation of V5/MT/MST bilaterally, the superior temporal sulcus bilaterally and the left intraparietal sulcus to causal relative to non-causal events. Directing attention to the causal nature of the stimuli had no significant effect on the neural processing of the causal events. These results support theories of causality suggesting that the perception of elementary mechanical causality events is automatically processed by the visual system. PMID- 11726786 TI - Impaired size processing for single objects after hemispatial neglect. AB - Some patients with hemispatial neglect show deficits in horizontal size perception. Most previous studies investigating this effect required the relative comparison of two horizontal stimuli. We examined whether the effect would also be present for single stimuli which would reflect an impairment in the computation of absolute horizontal length. Ten neglect patients, five with and five without hemianopia, and two control groups were asked to verbally judge the length of horizontal lines varying in length (3, 3.5, 4 inches) and spatial location (extreme left to extreme right). Three of the ten neglect patients judged a single object on the left as significantly smaller than a single object on the right, thus demonstrating a size processing deficit. This deficit was unrelated to hemianopia. PMID- 11726787 TI - Regulation of caspases and XIAP in the brain after asphyxial cardiac arrest in rats. AB - The aim of this study was to determine whether hypoxic-ischemia from asphyxial cardiac arrest activates brain caspases-1 and -3, and the anti-apoptotic protein, XIAP. Asphyxial cardiac arrest in rats was used to induce hypoxic-ischemia. A pan caspase inhibitor (zVAD) was given in the treatment group. At 72 h after reperfusion, caspase-3 and XIAP expression were present in multiple vulnerable brain regions, whereas caspase-1 was predominantly found in the CA1 hippocampus. zVAD significantly reduced expression of caspases and XIAP and the number of ischemic neurons in the CA1 hippocampus while neurological deficit scores were improved. We conclude that hypoxic-ischemia increases caspases-1 and-3, and XIAP expression. Treatment with zVAD significantly decreases caspase and XIAP expression in these brain regions and improves neurological outcome. PMID- 11726788 TI - Interleukin-1beta stimulates galectin-9 expression in human astrocytes. AB - Galectin-9 is an eosinophil chemoattractant produced by activated T lymphocytes. We have addressed expression of galectin-9 in normal human astrocytes in culture. Expression of galectin-9 mRNA and protein were examined by reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescent staining. Interleukin-1beta (IL-1beta) was found to enhance the galectin-9 expression in time- and concentration-dependent manners. Galectin-9 protein was detected in the membrane fraction, 105 000 x g precipitate, and immunofluorescent staining revealed diffuse cellular and perinuclear distributions. Dexamethasone pretreatment almost completely suppressed the production. We conclude that astrocytes produce galectin-9 in response to the stimulation with IL-1beta, and this may contribute to inflammatory reactions in the CNS. PMID- 11726789 TI - A new locus on chromosome 19 linked with late-onset Alzheimer's disease. AB - Alzheimer's disease (AD) is a complex neurodegenerative disorder, characterized by cognitive decline and distinctive neuropathology. APOE 4 and APOCI A on chromosome 19 are risk factors for late-onset disease. Using large extended families with multiple siblings affected, we have identified several microsatellite markers which are also linked with late- onset Alzheimer's disease. These microsatellites are distal from the apolipoprotein cluster on chromosome 19. It is likely that multiple genes will be involved with late-onset disease, either as risk factors or as causative agents. PMID- 11726790 TI - Effects of cerebral ischemia in mice lacking DNA methyltransferase 1 in post mitotic neurons. AB - DNA methylation is important for controlling gene expression and is catalyzed by DNA methyltransferase (Dnmt1) an enzyme abundant in brain. We recently demonstrated that mice expressing reduced levels of Dnmt1 are protected from cerebral ischemia. Here, we used the cre/loxP system to produce conditional mutants that lack Dnmt 1 in postmitotic neurons of the postnatal brain. We demonstrate that animals heterozygous for the conditional allele (Dnmt11lox/+) have significantly smaller infarcts following 1 h middle cerebral artery occlusion/reperfusion compared to their wildtype litters. Surprisingly, mice with a deletion of Dnmt1 in post-mitotic neurons (Dnmt11lox/c) were not protected. In conclusion, we demonstrate that reduced levels of Dnmt1, but not its absence, in post-mitotic neurons protect from ischemic brain injury. PMID- 11726791 TI - Cortical cholinergic decline parallels the progression of Borna virus encephalitis. AB - Borna disease virus (BDV)-induced meningoencephalitis is associated with the dysfunction of the cholinergic system. Temporal development of this cholinergic decline during pre-encephalitic and encephalitic stages of BDV infection remains however elusive. Changes in choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities were therefore determined in the cerebral cortex, hippocampus, striatum, amygdala and cholinergic basal forebrain nuclei (ChBFN) of rats infected with BDV. Immunocytochemistry for ChAT and vesicular acetylcholine transporter (VAChT) was employed to identify morphological consequences of BDV infection on cholinergic neurons. Whereas both ChAT and AChE activities changed only slightly under pre-encephalitic conditions, the encephalitic stage was characterized by a significant decrease of ChAT activity in the cerebral cortex, horizontal diagonal band of Broca (hDBB), hippocampus and amygdala concomitant with a marked reduction of AChE activity in the cerebral cortex, hDBB and hippocampus. The striatum and medial septum remained unaffected. ChAT and VAChT immunocytochemistry revealed prominent axonal degeneration in affected cortical and limbic projection areas of ChBFN. In summary, our data indicate progressive deterioration of forebrain cholinergic systems that parallels the progression of BDV encephalitis. PMID- 11726792 TI - Adenosine A2A, 5-HT1A and 5-HT7 receptor in neonatally pregnenolone-treated rats. AB - Steroid hormones synthesized in the brain, called 'neurosteroids', modulate neuronal activity. We treated neonatal rats with a main precursor of the neurosteroidogenesis, pregnenolone, and examined adenosine A2A receptor, 5- hydroxytryptamine (5-HT)1A and 5-HT7 receptor densities in the front-parietal cortex in juvenile and adult rats. In receptor binding assay using [3H]CGS21680 and [3H]8-OH-DPAT, it was shown that neonatal pregnenolone-treatment induced a significant decrease in the adenosine A2A receptor density with no significant effects on the 5-HT1A and 5-HT7 receptor densities. PMID- 11726793 TI - Event-related potential correlates of sound duration: similar pattern from birth to adulthood. AB - The effects of sound duration on event-related potentials (ERP) were studied in newborns and adults. Increasing tone duration from 200 to 300 ms led to the enhancement of the N2 peak amplitude, whereas two peaks became distinguishable in the N2 response elicited by 400 ms long tones. The sound-duration related ERP changes most likely reflect contribution from the sustained potential, although the observed results can also be explained by assuming the elicitation of a sound duration sensitive frontocentrally negative ERP component (duration-sensitive N2; DN2). The pattern of duration-related changes observed in newborn infants was very similar to that in adults, regardless of the structural differences between adult and infant ERPs. The results suggest that sound duration is processed already at birth in a similar way as in adulthood. PMID- 11726794 TI - Experimental nerve compression and upregulation of CPON in DRG. AB - Expression of C-terminal flanking peptide of neuropeptide Y (CPON) in DRG and cell proliferation (incorporation of BrdU) in sciatic nerve of rats following chronic nerve compression (silicone tubes with different internal diameters) was studied by immunocytochemistry. An increased number of CPON-positive neurons and cells incorporating BrdU was induced on the compressed side, most pronounced when a tight tube was used, while no cells expressed CPON or BrdU in intact nerves. The increase was transient and declined with time. Nerve compression induces transient cell proliferation in the nerve and expression of CPON in nerve cell bodies, but this is of a lesser magnitude than those following nerve transection. PMID- 11726795 TI - 17beta-Estradiol increases, aging decreases, c-Fos expression in the rat accessory olfactory bulb. AB - In the present paper we investigated the c-Fos immunoreactivity in the accessory olfactory bulb (AOB) of juvenile, adult and old rats of both sexes, as well as the effect of 17beta-estradiol on the expression of this immediate early gene. Basal c-Fos expression in the olfactory bulb decreased with age, and estrogen treatment caused an increase in the number of neurons expressing c-Fos in the AOB. The results indicate that both aging and estrogen have roles in the ability of neurons to co-ordinate genetic activity. Our observations may explain the decrease in age-related changes of brain plasticity, and provide data for the understanding of hormonally regulated neuronal plasticity. PMID- 11726796 TI - Delta9 -tetrahydrocannabinol increases brain temperature and inverts circadian rhythms. AB - Delta9-tetrahydrocannabinol (THC) has been shown to protect against focal and global ischemia. Hypothermia is thought to be one mechanism for this protection. These observations are important since brain hyperthermia is known to increase ischemic damage while hypothermia is protective. To establish the effect of THC on brain and body core temperature, brain and body temperature probes were inserted for chronic temperature monitoring (n = 20). THC treated groups were administered THC at either low (0.1 mg/kg) or high (10 mg/kg) dose for 1 week. Brain temperature was recorded during this period and for 1 week following the discontinuation of THC. Chronic administration of THC at either dose increased brain temperature (p < 0.0001) but did not significantly change body core temperature (p = 0.4767) in the freely moving rat. PMID- 11726797 TI - Subthalamo-pallido-striatal axis: a feedback system in the basal ganglia. AB - Systemic administration of a dopamine D2 receptor blocker, haloperidol, but not vehicle, significantly increased the number of c-Fos-immunoreactive neurons in the globus pallidus (GP) in rats. Dual immunohistochemistry, a combination of c Fos immunohistochemistry and retrograde tracing experiments with cholera toxin B (ChB), revealed that a subset of the c-Fos-immunoreactive GP neurons was pallidostriatal feedback neurons. Lesioning of the subthalamic nucleus (STN) by local injection of ibotenic acid inhibited the haloperidol-induced c-Fos expression in the GP neurons, suggesting that the activation of GP neurons is a result of increased excitatory drives from the STN. Therefore, the present findings are evidence of the existence of the subthalamo-pallido-striatal axis as a feedback system in the internal circuits of the basal ganglia. PMID- 11726798 TI - Acidic fibroblast growth factor (FGF-1) in the anterior horn cells of ALS and control cases. AB - The expression and localization of acidic fibroblast growth factor (aFGF; FGF-1) were examined in the spinal cord of patients with amyotrophic lateral sclerosis (ALS) and controls by reverse transcription-polymerase chain reaction (RT-PCR) method and immunohistochemistry. The RT-PCR experiments demonstrated that aFGF amplification products were clearly detected in all control cases but could be scarcely seen in ALS patients. aFGF immunoreactivity was detected in the anterior horn cells of the spinal cord. Double immunostaining for aFGF and choline acetyltransferase revealed that the majority (95.9%) of cholinergic neurons expressed aFGF. In ALS cases, the number and the staining intensity of aFGF positive neurons were markedly decreased. These results suggest that aFGF is involved in ALS pathology. PMID- 11726799 TI - Kinesthetic illusion of wrist movement activates motor-related areas. AB - We used positron emission tomography (PET) to test the hypothesis that illusory movement of the right wrist activates the motor-related areas that are activated by real wrist movements. We vibrated the tendons of the relaxed right wrist extensor muscles which elicits a vivid illusory palmar flexion. In a control condition, we vibrated the skin surface over the processes styloideus ulnae, which does not elicit the illusion, using the identical frequency (83 Hz). We provide evidence that kinesthetic illusory wrist movement activates the contralateral primary sensorimotor cortices, supplementary motor area (SMA) and cingulate motor area (CMA). These areas are also active when executing the limb movement. PMID- 11726800 TI - Somatotopic organization of the medial wall of the cerebral hemispheres: a 3 Tesla fMRI study. AB - Somatotopic organization of the human medial wall of the cerebral hemispheres was studied using functional MRI conducted at high field strength (3 T) with fine spatial resolution ( approximately 2 mm). Healthy subjects performed paced, repetitive movements of the fingers and toes. Within the supplementary motor area (SMA), two regions were identified: finger movements activated a region rostral and superior to that for toe movements. Two activation foci were also identified in the cingulate motor area: toe movements activated a region rostral and ventral to that activated by finger movements. All foci were located between the anterior and posterior commissures. Our results confirm previous human and non-human primate studies regarding the rostral-caudal organization of the SMA and CMA. The dorsal-ventral organization of the CMA, however, appears to be divergent from results derived from cortical stimulation studies conducted in non-human primates. PMID- 11726801 TI - Early visual processing deficits in schizophrenia: impaired P1 generation revealed by high-density electrical mapping. AB - Integrity of early visual sensory processing in schizophrenia was assessed using the well characterized P1 and N1 components of the visual evoked potential (VEP) as our dependent measures. VEPs were recorded in response to successively less fragmented line drawings of common objects. P1 amplitudes were significantly reduced across all stimulus conditions for patients versus controls. Further, this decrement was relatively greater at parieto-occipital than occipito-temporal electrode sites. No differences in N1 amplitude were found. The finding of P1 deficits in patients, particularly over dorsal scalp, supports the view that schizophrenia is associated with impairment of early dorsal visual stream processing. On the other hand, the finding of normal N1 amplitudes in patients suggests that early stages of ventral stream processing may be relatively more intact. These results imply that the cognitive impairment seen in schizophrenia is not just due to deficits in higher order aspects of cognition but also encompasses significant deficits in early sensory processing. PMID- 11726802 TI - Effects of acute hypotension on neuronal activity in the medial vestibular nuclei of rats. AB - The role of peripheral vestibular receptors in acute hypotension was investigated in anesthetized rats. In animals with intact labyrinths, acute hypotension induced by either i.v. infusion of sodium nitroprusside or hemorrhage produced excitation of electrical activity in two-thirds of type I neurons and inhibition in two-thirds of type II neurons recorded in the medial vestibular nuclei. In unilaterally labyrinthectomized animals, two-thirds of type I neurons ipsilateral to the lesion showed an inhibitory response, and two-thirds of contralateral type I neurons showed an excitatory response after the induction of acute hypotension. The response patterns of type II neurons were opposite to those of type I neurons. These results suggest that blood flow changes are detected by peripheral vestibular receptors, and that this might suggest a mechanism for control of blood pressure. PMID- 11726803 TI - Input minimization: a model of cerebellar learning without climbing fiber error signals. AB - The cerebellum is critical for motor learning. Current cerebellar learning models follow the Marr/Albus paradigm, in which climbing fibers provide error signals that shape plastic synapses between parallel fibers and Purkinje cells. However, climbing fibers have slow and largely random discharge, and seem unlikely to provide error signals with resolution sufficient to guide cerebellar learning. Parallel fibers carry error signals and could direct the plasticity of their own synapses, but the error signals are carried along with other signals. This report presents the new input minimization (InMin) model, in which Purkinje cells reduce error by minimizing their overall parallel fiber input. The slowly, randomly firing climbing fiber provides only synchronization pulses. InMin offers an alternative that can unify cerebellar findings. PMID- 11726804 TI - Photic regulation of melatonin in rat retina and the role of proteasomal proteolysis. AB - Several investigations have shown that illumination at night reduces melatonin level in the mammalian pineal, but the effect of night illumination on the retina is not known. In this study retinas were cultured in a flow-through apparatus and then were exposed to light at ZT 18. Light exposure reduced melatonin levels to the daytime level within 30 min. The reduction of melatonin levels was due to a rapid decrease in the activity of the enzyme AA-NAT; AA-NAT mRNA levels were not affected by illumination. Pre-incubation with lactacystin (25 microM) prevented light-induced reduction of AA-NAT activity and melatonin levels. These results demonstrate that melatonin levels in the mammalian retina are affected by light exposure at night, via proteosomal proteolysis of AA-NAT. PMID- 11726805 TI - Phosphorylation of amyloid-beta at the serine 26 residue by human cdc2 kinase. AB - The amyloid-beta (Abeta) peptide has been implicated in the pathology of Alzheimer's disease (AD). Using an antisense peptide approach a novel interaction between Abeta and the human cdc2 kinase was identified. The Abeta 1-42, 1-40 and 25-35 peptides were shown to be substrates for the cdc2 kinase and phosphorylated on the Serine 26 residue. Phosphorylated Abeta (pSAbeta) was found in extracts from NT-2 neurons and AD brain. In NT-2 neurons the levels of pSAbeta were increased in the presence of exogenous Abeta and this increase was prevented by a cdc2 protein kinase inhibitor, olomoucine, that also prevented Abeta cytotoxicity. The results from this study suggest that Abeta phosphorylation by cdc2 could play a role in the brain pathology of AD. PMID- 11726806 TI - Modulation of spinal cord excitability by subthreshold repetitive transcranial magnetic stimulation of the primary motor cortex in humans. AB - Repetitive transcranial magnetic stimulation (rTMS) allows the modulation of intra-cortical excitability and may therefore affect the descending control of spinal excitability. We applied rTMS at subthreshold intensity and 1 Hz frequency for 10 min to the left primary motor cortex representation of the flexor carpi radialis muscle (FCR) in 10 subjects and assessed the H and M responses to median nerve stimulation before and after the rTMS. Following rTMS, H wave thresholds significantly reduced by approximately 20%. Maximal H but not M wave amplitude significantly increased over the baseline, so that H/M amplitude ratio was increased by 41%. Sham stimulation did not induce any noticeable change in M or H waves. Slow rTMS might facilitate monosynaptic spinal cord reflexes by inhibiting the cortico-spinal projections modulating spinal excitability. PMID- 11726807 TI - Sound alters visual evoked potentials in humans. AB - When a single flash is accompanied by two auditory beeps, the single flash is perceived as two flashes. We investigated whether this crossmodal influence on visual perception occurs at the level of the modality-specific visual pathway or later. We compared the visual evoked potentials (VEPs) in the presence and absence of sound. Activity was modulated extensively and with short latency in trials in which an illusory flash was perceived. In addition, the brain potentials for the illusory flash were qualitatively very similar to those for a physical flash, suggesting that the same mechanism underlies the percept of both illusory and physical flashes. These results suggest that the activity in the visual cortex can be modulated by sound. This implication challenges the general belief that the visual cortical processing is independent of other modalities. PMID- 11726808 TI - Endothelin-1 activates ET(A) receptors to increase intracellular calcium in model sensory neurons. AB - Endothelin-1 (ET-1) induces endothelin-A (ETA) receptor-mediated pain and selective excitation of nociceptors. Here we studied ET-1-induced changes in intracellular calcium (Ca2+in) in Fura-2 loaded mouse neuroblastoma-rat dorsal root ganglion hybrid cells (ND7/104). ET-1 (1-400 nM) induced concentration dependent, transient increases in Ca2+in, probably of intracellular source. Responses to repeated application declined with increasing ET-1 concentration, implying receptor desensitization. Treatment of cells with the selective ETA receptor antagonist, BQ-123, produced a dose-dependent inhibition of the response that was 20% of ET-1 alone (IC50 = 20 nM, KI = 7 nM). No inhibition of the calcium response was observed with the selective ETB antagonist, BQ-788 (10-1000 nM). These results demonstrate that ET-1 induces dose- and ETA receptor-dependent release of Ca2+in in nociceptor-like neurons, and permit further examination of the pathways that underlie ET-1-induced pain signaling. PMID- 11726809 TI - Does post-movement beta synchronization reflect an idling motor cortex? AB - After the completion of a voluntary movement, a synchronization of cortical beta rhythms is recorded over the contralateral central region, which is assumed to reflect the termination of the motor command. In order to test this hypothesis, we compared in eight healthy subjects the synchronization of EEG beta rhythms following active and passive index extension. The passive movement was also performed after deafferentation by ischaemic nerve block in three subjects. Beta synchronization was present in all subjects after both active and passive movements, and disappeared under ischaemia in all three subjects. Post-movement beta synchronization can not solely be explained by an idling motor cortex. It may also, at least in part, reflect a movement-related somatosensory processing. PMID- 11726810 TI - Semantic acquisition without memories: evidence from transient global amnesia. AB - Transient global amnesia (TGA), characterised by a profound anterograde amnesia, is a model of interest to study the acquisition of novel meanings independent of episodic functioning. Three patients were tested during a TGA attack, two in the early recovery phase and the third during the acute phase of TGA, with a semantic priming task involving a restructuring process of conceptual knowledge. During TGA, all patients demonstrated priming effects. Results obtained the day after the episode with the same task showed that these effects persisted at least one day. Episodic memory seems not to be critical for the formation of novel connections among unrelated semantic representations, in accordance with Tulving's model of memory, i.e. episodic memory is not necessary for the acquisition of semantic information. PMID- 11726811 TI - Tissue distribution and neuroprotective effects of citrus flavonoid tangeretin in a rat model of Parkinson's disease. AB - Neuroprotective effects of a natural antioxidant tangeretin, a citrus flavonoid, were elucidated in the 6-hydroxydopamine (6-OHDA) lesion rat model of Parkinson's disease (PD), after bioavailability studies. Following the chronic oral administration (10 mg/kg/day for 28 days), significant levels of tangeretin were detected in the hypothalamus, striatum and hippocampus (3.88, 2.36 and 2.00 ng/mg, respectively). The levels in the liver and plasma were 0.59 ng/mg and 0.11 ng/ml respectively. Unilateral infusion of the dopaminergic neurotoxin, 6 hydroxydopamine (6-OHDA; 8 microg), onto medial forebrain bundle significantly reduced the number of tyrosine hydroxylase positive (TH+) cells in the substantia nigra and decreased striatal dopamine content in the vehicle treated rats. Sub chronic treatment of the rats with high doses of tangeretin (20 mg/kg/day for 4 days; p.o.) before 6-OHDA lesioning markedly reduced the loss of both TH+ cells and striatal dopamine content. These studies, for the first time, give evidence that tangeretin crosses the blood-brain barrier. The significant protection of striato-nigral integrity and functionality by tangeretin suggests its potential use as a neuroprotective agent. PMID- 11726812 TI - Embryonic cell grafting for the treatment of peripheral nervous system diseases. PMID- 11726813 TI - Assembling the vertebrate retina: global patterning from short-range cellular interactions. PMID- 11726814 TI - BK virus in solid organ transplant recipients: an emerging syndrome. AB - BK virus is a human polyomavirus associated with a range of clinical presentations from asymptomatic viruria with pyuria to ureteral ulceration with ureteral stenosis in renal transplant patients or hemorrhagic cystitis in bone marrow transplant recipients. Infection of renal allografts has been associated with diminished graft function in some individuals. Fortunately, however, the majority of patients with BK virus infections are asymptomatic. The type, duration, and intensity of immunosuppression are major contributors to susceptibility to the activation of BK virus infection. Histopathology is required for the demonstration of renal parenchymal involvement; urine cytology and viral polymerase chain reaction methods are useful adjunctive diagnostic tools. Current, treatment of immunosuppressed patients with polyomavirus viruria is largely supportive and directed toward minimizing immunosuppression. Improved diagnostic tools and antiviral therapies are needed for polyomavirus infections. PMID- 11726815 TI - Posttransplantation Diabetes Mellitus in FK-506-Treated Renal Transplant Recipients: Analysis of Incidence and Risk Factors. Transplantation 2001; 72: 1655. PMID- 11726816 TI - Liver Transplantation in Adults Coinfected With HIV. Transplantation 2001; 72: 1684. PMID- 11726817 TI - "Accomodated" pig endothelial cells promote nitric oxide-dependent Th-2 cytokine responses from human T cells. AB - BACKGROUND: Cardiac and renal allo- and xenografts can become naturally resistant to vascular rejection. Understanding this process of "accommodation" would enhance our understanding of vascular inflammatory responses and have implications for immune manipulation and tolerance induction. A feature of these grafts is infiltration by leukocytes secreting a Th-2 pattern of cytokines. METHODS: HLA-DR-1-transfected, immortalized porcine endothelial cells (IPEC) were incubated with polyclonal human immunoglobulin G (IgG) for 6 days before incubation with purified human CD4+ T cells. RESULTS: IgG-incubated IPEC stimulated a normal proliferative response from alloreactive T cells. However, interferon (IFN)-gamma levels were significantly reduced, whereas interleukin (IL)-5 and IL-10 were maintained at levels equivalent to those stimulated by control IPEC. Cognate interaction between T cells and IPEC was not required for this effect, because IgG-incubated, MHC-class II-negative IPEC caused reduced IFN gamma secretion during a response to human Epstein-Barr virus-transformed B cells. Experiments with the nitric oxide (NO) donor, (z)-1-2-[2-Aminoethyl)-N-(2 ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA-NO), and the NO synthase inhibitor, NG-monomethyl-L-arginine.monoacetate (L-NMMA), showed that NO released by the IgG-incubated IPEC was actively involved in the development of this phenotype. CONCLUSIONS: These data suggest a novel, IgG-mediated, NO-dependent mechanism by which endothelial cells (EC) influence T cell responsiveness and that the Th-2 cytokine skewing seen in "accommodated" grafts may be a secondary phenomenon, resulting from the T-EC interactions. PMID- 11726818 TI - Myocyte function and [Ca 2+ ]i homeostasis during early allogenic heart transplant rejection. AB - BACKGROUND: Recent studies from our laboratory have demonstrated that in vivo contractile function of rejecting mouse heterotopic abdominal heart allografts 5 days after transplantation is depressed to 40% of that of syngenic controls, and that this depression of function is prevented by the nitric oxide synthase (NOS) inhibitor NG-monomethyl-l-arginine. However, the mechanisms of altered myocyte function caused by nitric oxide production in this setting are not established. METHODS: We measured intracellular calcium concentration ([Ca2+]i) transients (fluo-3, confocal microscopy), fractional shortening (video motion), and L-type Ca2+ currents (whole-cell patch clamp) 5 days after transplantation in ventricular myocytes freshly isolated from syngenic (Balb/C into Balb/C) and allogenic (Balb/C into C3H) transplants. RESULTS: L-type Ca2+ currents, [Ca2+]i transient amplitudes, and fractional shortening did not differ between nonrejecting, syngenic and rejecting, allogenic transplants. Catecholamine responsiveness as analyzed by the change in the peak [Ca2+]i transient induced by 100 nM isoproterenol was also similar. Superfusion with l-arginine, an NOS substrate, caused decreased shortening with no change in [Ca2+]i transients in allogenic myocytes, but had no effect in syngenic myocytes. CONCLUSIONS: Depressed contractile function of rejecting allogenic heart transplants in vivo appears to be caused in part by an NOS-dependent decrease in myofilament Ca2+ sensitivity. PMID- 11726819 TI - Highly disparate xenogeneic skin graft tolerance induction by fetal pig thymus in thymectomized mice: Conditioning requirements and the role of coimplantation of fetal pig liver. AB - BACKGROUND: Highly disparate xenogeneic pig skin graft tolerance and efficient repopulation of mouse CD4+ T cells are achieved in thymectomized (ATX) B6 mice that receive T cell and natural killer (NK) cell depletion by injection of a mixture of monoclonal antibodies (mAbs) (GK1.5, 2.43, 30-H12, and PK136) on days 6, -1, +7, and +14 and 3 Gy total body irradiation (TBI) followed by implantation of fetal pig thymus/liver (FP THY/LIV) grafts on day 0. The requirements for each treatment in this model to achieve pig skin graft tolerance have not previously been defined. Therefore, we performed a series of experiments to address the role of each treatment in achieving maximal skin graft tolerance. METHODS: Peripheral mouse CD4+ T-cell repopulation and pig skin graft survival were followed in this pig-to-mouse model in which recipient B6 mice were treated with modified regimens that omitted thymectomy, 3 Gy TBI, anti-Thy1.2, and anti-NK1.1 mAbs, injection of a mixture of mAbs on day +14, or coimplantation of FP LIV, respectively. RESULTS: Prolongation but not permanent survival of donor MHC-matched pig skin grafts was observed in euthymic B6 mice that received T and NK cell depletion, 3 Gy TBI, and 7 Gy thymic irradiation and FP THY/LIV in the mediastinum, suggesting that full xenogeneic tolerance was not achieved in euthymic mice. However, after grafting FP THY alone to ATX B6 mice treated either with the "standard" regimen, or with a conditioning regimen that omitted all components of the conditioning regimen except treatment with anti-CD4 and anti-CD8 mAbs, efficient peripheral repopulation of mouse CD4+ T cells and long-term donor MHC-matched pig skin graft acceptance were observed. CONCLUSIONS: Highly disparate xenogeneic pig skin graft tolerance can be achieved by grafting FP THY alone in anti-CD4 and anti-CD8 mAb treated ATX B6 mice, but not in euthymic B6 mice. Additional treatment of ATX recipient mice with anti-Thy1.2 and NK1.1 mAbs and 3 Gy TBI is not essential for donor pig skin graft tolerance induction. PMID- 11726820 TI - Dependence of murine obstructive airway disease on CD40 ligand. AB - BACKGROUND: Human lung transplantation carries a poor prognosis because of chronic rejection in the form of obliterative bronchiolitis syndrome (OBS). Using the mouse model of heterotopic tracheal transplantation, we examined the role of costimulation in the allograft rejection that characterizes obstructive airway disease (OAD). METHODS: C57BL/6 or BALB/c tracheae were implanted into wild-type control, CD28-/-, muMT (B-cell deficient), or CD40L-/- recipient mice. Grafts were explanted from 7 to 42 days posttransplantation and evaluated. RESULTS: Thickening of the basement membrane and a decrease in patent luminal area were first noted at 2 weeks in wild-type allogeneic trachea recipients and to a slightly lesser degree in CD28-/- recipients. In contrast, CD40L-/- recipient mice showed no evidence of cellular infiltrates or fibrosis in transplanted tracheae. To determine whether CD40L interacted with host or donor CD40, CD40 deficient tracheae were transplanted into CD40L+/+, CD40+/+ wild-type mice. Wild type mice rejected CD40-/- tracheae. Tracheae were transplanted into B-cell deficient mice to determine the role of B-cell CD40 in chronic pulmonary allograft rejection. The OAD reaction was identical in wild-type and B-cell deficient mice. CONCLUSIONS: Development of OAD in the mouse trachea transplant model is primarily dependent on CD40L and is relatively CD28 independent. The ability of mice to reject CD40-/- tracheae demonstrated that host, not donor, CD40 is required for rejection. Furthermore, the ability of B-cell-deficient mice to reject allogeneic tracheae demonstrated that B-cell CD40-mediated responses are not required for the development of OAD. PMID- 11726822 TI - Donor brain death reduces survival after transplantation in rat livers preserved for 20 hr. AB - BACKGROUND: Eighty percent of donor organs come from donors who have suffered brain trauma (brain-dead donors). This unphysiological state alters the hemodynamic and hormonal status of the organ donor. This can cause organ injury, which has been suggested to alter the immunological or inflammatory status of the organ after transplantation, and may lead to increased sensitivity of the organ to preservation/transplantation injury. In this study we asked the question: does brain death cause injury to the liver that decreases successful liver preservation? METHODS: The rat liver transplant model was used to compare survival in rats receiving a liver from a brain-dead donor versus a non-brain dead donor. Brain death was induced by inflation of a cranially placed balloon catheter. The rats were maintained normotensive with fluid infusion for 6 hr. The livers were flushed with University of Wisconsin (UW) solution and immediately transplanted or cold stored for 20 hr before transplantation. RESULTS: Recipient survival with immediately transplanted livers or those stored for 20 hr was 100% with livers from non-brain-dead donors. However, survival decreased when livers were procured from brain-dead donors. Survival was 75% (6/8) when storage time was 0 hr and 20% (2/10) when the liver was cold stored for 20 hr before transplantation. CONCLUSION: This study shows that brain death induces alterations in the donor liver that make it more sensitive to preservation/reperfusion injury than livers from donors without brain death. The mechanism of injury to the liver caused by brain death is not known. Because most livers used clinically for transplantation come from brain-dead donors, it is possible that poor function of these livers is due to the intrinsic condition of the donor organ, more than the quality of the preservation. Methods to treat the brain-dead donor to improve the quality of the liver may be needed to allow better preservation of the organ and to give better outcome after liver transplantation. PMID- 11726821 TI - Effect of ischemia-reperfusion injury on the microcirculation of the steatotic liver of the Zucker rat. AB - BACKGROUND: Much discussion has been focused on the use of steatotic livers for transplantation due to the prevalence of steatosis in the potential donor liver pool (1). The aim of this study was to investigate the possibility that the microcirculation of steatotic liver is more sensitive to the ischemia-reperfusion (IR) injury than normal liver. METHODS: The left liver lobe of obese (n=9) and lean Zucker rats (n=9) were subjected to 40 min of warm ischemia followed by 60 min of reperfusion. Fluorescent probes rhodamine 123 (Rh123), bisbenzimide (Bis), and rhodamine 6G (Rh6G) were administered for the identification by intravital fluorescence microscopy (IVFM) of mitochondrial membrane potential, hepatocyte nuclei and leukocytes, respectively before hepatic ischemia and at 15, 30, 45, and 60 min after reperfusion. Blood samples were obtained before and after 60 min of reperfusion. Liver tissue was taken at the end of experiment for histological analysis. RESULTS: The liver of the obese rats showed prominent macro- and microvesicular fatty changes (MAFC and MIFC) and hepatocyte swelling. Under IVFM, the obese animals had significantly wider hepatic cords (23.1+/-0.8 microm) than the lean ones (15.9+/-0.5 microm) (P<0.01), whereas no significant difference in sinusoidal diameters was noted. The number of functional sinusoids significantly decreased after 30 min of reperfusion in both groups but no significant change was noted in the nucleus count throughout the experiment. Rh123 fluorescence intensity dropped significantly in the obese group after 60 min of reperfusion but not in the lean rats. Leukocyte adherence showed a significant rise after reperfusion in both groups. Plasma AST and ALT levels were 40- and 24-fold higher respectively for the obese animals after IR compared with their preischemic values, whereas the corresponding increase were 4.2- and 3.4-fold for the lean animals, respectively. CONCLUSIONS: Our results indicate that the liver of the obese Zucker rat is steatotic and presents with an abnormal microcirculation manifested by a reduced sinusoidal density. IR led to significantly greater hepatic injury in the steatotic than in the normal liver. This injury was accompanied by a significant reduction in the functional sinusoidal density and mitochondrial membrane potential as assessed by Rh123-associated fluorescence in the steatotic liver. In conclusion, the increased sensitivity of the steatotic liver to IR injury would appear to involve both alterations in blood flow in the microcirculation and to cellular changes. PMID- 11726823 TI - Multicenter survey of daclizumab induction in simultaneous kidney-pancreas transplant recipients. AB - We report an experience with 71 simultaneous kidney-pancreas transplant (SKPT) recipients receiving daclizumab induction in combination with tacrolimus (TAC), mycophenolate mofetil (MMF), and steroids. The mean follow-up time was 5.9+/-2.5 (SD) months (range 0.5-11 months). The study population included 47 males (65%) and 24 females (35%) with a mean age of 40+/-8 years. The mean pretransplant duration of diabetes and dialysis were 25+/-8 and 1.5+/-0.9 years (34 hemodialysis, 16 peritoneal dialysis), respectively. Mean HLA match was 1.2+/ 1.5, with one patient receiving a second transplant. The mean cold ischemic times for the kidney and the pancreas were 15+/-5 and 16+/-4 hr, respectively. Six month patient, kidney, and pancreas graft survival and rejection rates were 97, 96, 93, and 35%, respectively. There were two deaths, one due to fungal infection and the other due to a cardiac event. There were three kidney graft losses, two immunological, and one death with function. Of the five pancreas graft losses, two were due to infection, one immunological, one thrombosis, and one death with function. The patient population was then stratified according to the number of daclizumab doses: 4-5 doses (n=45) or 1-3 doses (n=26). There were no differences in patient and kidney graft survival rates, 98 vs. 96%, and 92 vs. 92%, respectively. However, there was a trend toward improved pancreas graft survival in the group receiving 4-5 doses (96%) compared with 1-3 doses (85%), P=0.07. Although more patients receiving 1-3 doses had rejection (54%) than patients receiving 4-5 doses (24%), there was no dose response relationship between the total number of doses or the adjusted total mg/kg dose and time to rejection. All patients with functioning grafts have good renal and pancreas allograft function at 6 and 12 months. The overall incidence of major infection was 27% and there were no differences in the incidence of infection between the two groups. No major adverse events were attributed to daclizumab use. In conclusion, excellent short-term outcomes were noted in this retrospective, multicenter survey of initial experience with daclizumab induction in combination with TAC, MMF, and steroids in SKPT recipients. Optimal dosing strategies for SKPT recipients remain to be determined. PMID- 11726824 TI - Leukemia after liver transplant. AB - INTRODUCTION: Acute leukemia is rare after solid organ transplantation. METHODS: Review of data on 3 patients with acute leukemia identified among 1365 who underwent liver transplantation at our center, and a review of the literature. RESULTS: In patient 1, AML-M4 developed 19 months after transplant for cryptogenic cirrhosis. In patient 2, B cell acute lymphoid leukemia was diagnosed 10 months after liver transplant for fulminant hepatitis. Both patients had normal cytogenetics, and achieved complete remission with chemotherapy. In patient 3, acute monocytic leukemia-M3 with t(15;17) arose 18 months after transplant for hepatitis C cirrhosis. This patient received treatment with retinoic acid and chemotherapy, but developed a disseminated intravascular coagulation and died before completing therapy. No patient presented with chromosomal abnormalities commonly seen in secondary leukemia. The latency period to diagnosis after transplant was 10-19 months. CONCLUSIONS: Acute leukemia, although rare after liver transplantation, should be considered in the differential diagnosis of hematological complications. PMID- 11726825 TI - Impact of cytomegalovirus hyperimmune globulin on outcome after cardiothoracic transplantation: a comparative study of combined prophylaxis with CMV hyperimmune globulin plus ganciclovir versus ganciclovir alone. AB - BACKGROUND: Cytomegalovirus (CMV) disease was previously shown to be unaltered by a 28-day course of ganciclovir compared with placebo in seronegative recipients of hearts from seropositive donors (D+/R-). This study tests the hypothesis that a combination of ganciclovir plus CMV hyperimmune globulin (CMVIG) is more effective than ganciclovir alone for preventing acute CMV illness and its long term sequelae. METHODS: The study population receiving CMVIG (n=80) included 27 heart transplant recipients (D+/R-) and 53 heart-lung and lung transplant recipients (R+ and/or D+). Each group was matched with historical controls who underwent transplantation within the preceding 2-3 years. Outcome measures compared were as follows: 3-year incidence of CMV disease; fungal infection; acute rejection; survival; rates and severity of transplant coronary artery disease (in heart patients) defined by intimal thickness (ultrasound) and coronary artery stenosis (angiographic); and incidence and death from obliterative bronchiolitis defined by pathological criteria on endobronchial biopsy specimens (in heart-lung/lung patients). RESULTS: Patients treated with CMVIG had a higher disease-free incidence of CMV, lower rejection incidence, and higher survival rate compared with the patients treated with ganciclovir alone. The coronary artery intimal thickness and the prevalence of intimal thickening were lower in the patients receiving CMVIG. Heart-lung and lung transplant patients treated with CMVIG had lower incidences of obliterative bronchiolitis and death from obliterative bronchiolitis and longer survival compared with the patients treated with ganciclovir alone. CONCLUSIONS: CMVIG plus ganciclovir seems to be more effective that ganciclovir alone for preventing the sequelae of CMV infection. A prospective randomized study is required to confirm these observations. PMID- 11726826 TI - Stem cell transplantation eliminates alloantibody in a highly sensitized patient. AB - Highly sensitized patients are forced to stay on transplant waiting lists for many years and ultimately may never find a donor. Peripheral blood stem cell (PBSC) transplantation may provide a strategy to decrease host alloreactivity through the production of a chimeric state. We investigated alloreactivity and chimerism in a highly sensitized 40-year-old patient with sickle cell disease who underwent a nonradiation based conditioning regimen consisting of fludarabine, ATG, and high dose melphalan, for allogeneic stem cell transplant. Host monocytes and lymphocytes became donor in origin by day 14. PRA, initially 100% pretransplant, fell to 0 by day 263. Anti-red blood cells antibody became undetectable by day 152. The use of a new nonradiation-based conditioning regimen enabled successful engraftment of allogeneic donor PBSCs and the elimination of alloantibody. As new less toxic conditioning regimens are developed, PBSC transplantation might provide a new solution to allosensitization. PMID- 11726827 TI - Posttransplantation diabetes mellitus in FK-506-treated renal transplant recipients: analysis of incidence and risk factors. AB - BACKGROUND: The development of posttransplantation diabetes mellitus has a major impact on the quality of life and long-term outcome. METHODS: One hundred thirty nine patients without known glucose metabolism abnormalities and treated with FK 506, methylprednisolone, and mycophenolate mofetil/azathioprine were analyzed for incidence of and risk factors for developing impaired fasting glycemia (IFG) and diabetes mellitus (DM). RESULTS: Using the American Diabetes Association criteria, 15% developed IFG and 32% developed DM in the first year after transplantation. High trough levels of FK-506 during the first month after transplantation (especially >15 ng/ml) and high body mass index (BMI) were significant risk factors for IFG or DM. Patients with (steroid-treated) acute rejections in addition to high trough levels of FK-506 were most prone to develop DM, whereas high BMI conferred risk of developing IFG. Patients with posttransplantation glycemic abnormalities also had higher levels of serum triglycerides at the time of transplantation, but they needed a lower dose of FK 506 to obtain higher trough levels of FK-506, suggesting metabolic differences already present before transplantation. The only risk factor retained for persistent IFG or DM beyond the first year was a higher number of trough levels of FK-506 >15 ng/ml during the first month after transplantation. CONCLUSIONS: Induction with an FK-506 based immunosuppressive regimen resulted in a high incidence of glucose metabolism disorders in renal transplantation recipients. Higher trough levels of FK-506 during the first month, acute rejections, and higher BMI were the most obvious risk factors. PMID- 11726828 TI - Mycophenolate mofetil versus azathioprine in patients surviving the initial cardiac transplant hospitalization: an analysis of the Joint UNOS/ISHLT Thoracic Registry. AB - BACKGROUND: Although the randomized mycophenolate mofetil- (MMF) azathioprine (AZA) trial is likely applicable to cardiac transplantation in general, it was limited to select and usually larger cardiac transplant centers and suffered from substantial cross-over and failure of many patients to receive assigned treatment drug. METHODS: The Joint ISHLT/UNOS Thoracic Registry was analyzed for the effects of MMF versus AZA in patients 1) on a cyclosporine- (CsA) based immunosuppression protocol; 2) having survived long enough to be discharged from the transplant hospitalization. RESULTS: A total of 5599 patients (4942 CsA/AZA and 657 CsA/MMF) were included with no significant differences between the MMF and AZA groups in baseline characteristics with the exception of recipient age (50 vs. 47 years), donor age (29 vs. 28 years), ischemic time (3.0 vs. 2.9 hr), and pretransplant medical condition (more AZA patients in ICU, more MMF patients on VAD). Actuarial survival was greater in the MMF group compared to the AZA group in patients surviving the initial transplant hospitalization (1 year 96 vs. 93%, 3 years 91 vs. 86%, P=0.0012). This difference was confirmed in the logistic regression analysis of 3-year mortality showing a relative risk of 0.62 (P=0.011). CONCLUSIONS: These data provide independent support for the broad applicability of the positive results from the randomized MMF-AZA clinical trial in a substantially larger patient population and confirm improved survival in patients using mycophenolate mofetil compared to azathioprine late after cardiac transplantation. PMID- 11726829 TI - Results of simultaneous and sequential pediatric liver and kidney transplantation. AB - BACKGROUND: The indications for simultaneous and sequential pediatric liver (LTx) and kidney (KTx) transplantation have not been well defined. We herein report the results of our experience with these procedures in children with end-stage liver disease and/or subsequent end-stage renal disease. PATIENTS AND METHODS: Between 1984 and 1995, 12 LTx recipients received 15 kidney allografts. Eight simultaneous and seven sequential LTx/KTx were performed. There were six males and six females, with a mean age of 10.9 years (1.5-23.7). One of the eight simultaneous LTx/KTx was part of a multivisceral allograft. Five KTx were performed at varied intervals after successful LTx, one KTx was performed after a previous simultaneous LTx/KTx, and one KTx was performed after previous sequential LTx/KTx. Immunosuppression was with tacrolimus or cyclosporine and steroids. Indications for LTx were oxalosis (four), congenital hepatic fibrosis (two), cystinosis (one), polycystic liver disease (one), A-1-A deficiency (one), Total Parenteral Nutrition (TPN)-related (one), cryptogenic cirrhosis (one), and hepatoblastoma (one). Indications for KTx were oxalosis (four), drug-induced (four), polycystic kidney disease (three), cystinosis (one), and glomerulonephritis (1). RESULTS: With a mean follow-up of 58 months (0.9-130), the overall patient survival rate was 58% (7/12). One-year and 5-year actuarial patient survival rates were 66% and 58%, respectively. Patient survival rates at 1 year after KTx according to United Network of Organ Sharing (liver) status were 100% for status 3, 50% for status 2, and 0% for status 1. The overall renal allograft survival rate was 47%. Actuarial renal allograft survival rates were 53% at 1 and 5 years. The overall hepatic allograft survival rate was equivalent to the overall patient survival rate (58%). Six of seven surviving patients have normal renal allograft function, and one patient has moderate chronic allograft nephropathy. All surviving patients have normal hepatic allograft function. Six (86%) of seven sequentially transplanted kidneys developed acute cellular rejection compared with only two (25%) of eight simultaneously transplanted kidneys (P<0.04). CONCLUSIONS: Simultaneously transplanted kidneys were less likely to develop rejection than sequentially transplanted kidneys in this series. This did not have any bearing on patient or graft survival rates. Mortality correlated directly with the severity of United Network of Organ Sharing status at the time of kidney transplantation. Candidates for simultaneous or sequential LTx/KTx should be prioritized based on medical stability to optimize distribution of scarce renal allografts. PMID- 11726830 TI - Thymoglobulin induction decreases rejection in solitary pancreas transplantation. AB - BACKGROUND: Solitary pancreas transplants, both pancreas transplant alone (PTA) and pancreas after kidney (PAK), have higher rejection rates and lower graft survivals than simultaneous pancreas-kidney transplants (SPK). The aim of this study is to compare three different antibody induction regimens in solitary pancreas transplant recipients and to assess the role of surveillance pancreas biopsies in the management of these patients. METHODS: Solitary pancreas transplant recipients between 01/98 to 02/00 (n=29) received induction with either daclizumab (1 mg/kg on day 0, 7, 14), OKT 3 (5 mg/day x0-7), or thymoglobulin (1.5 mg/kg/day x0-10). Maintenance immunosuppression was similar for the three groups. All rejections were biopsy-proven either by surveillance/protocol or when clinically indicated. RESULTS: The 1-year graft survival was 89.3% overall and 91.7% in the thymoglobulin group. Thymoglobulin significantly decreased rejection in the first 6 months when compared with OKT3 or daclizumab (7.7 vs. 60 vs. 50%). Acute rejections were seen on surveillance biopsies in the absence of biochemical abnormalities in 40% of patients. CONCLUSIONS: Thymoglobulin induction regimen led to a low incidence of acute rejection and a high rate of graft survival in solitary pancreas transplants. In addition, surveillance biopsies were useful in the detection of early acute rejection in the absence of biochemical abnormalities. PMID- 11726831 TI - Steroid elimination 24 hours after liver transplantation using daclizumab, tacrolimus, and mycophenolate mofetil. AB - BACKGROUND: Corticosteroids have long been a cornerstone of orthotopic liver transplant (OLTx) immunosuppression. Newer, more potent, agents have successfully allowed for more rapid tapering and discontinuation of corticosteroids in OLTx recipients. We hypothesize that corticosteroids can be safely avoided after the first postoperative day (POD) using these newer agents. METHODS: Thirty adult OLTx recipients were prospectively enrolled in a randomized open-label, institutional review board-approved protocol. Fifteen patients (group A) received our standard regimen of tacrolimus, mycophenolate mofetil, and corticosteroids, and 15 patients (group B) received daclizumab, 2 mg/kg on POD 0 and 14, with tacrolimus, mycophenolate mofetil, and corticosteroids on POD 0 and 1 and then discontinuation. In both groups, mycophenolate mofetil was tapered off between 3 and 4 months after OLTx. Bone mineral densitometry was performed at 1, 3, and 6 months after OLTx. Quantitative hepatitis C virus (HCV) polymerase chain reaction was obtained at days 0, 7, 14, 21, and 28. Retransplant recipients, patients with autoimmune hepatitis, or status 1 or 2A patients were excluded. RESULTS: Patient and graft survival rates were 93% (group A) and 100% (group B) with mean follow up of 18 months. Patients in group B had more rejection diagnosed (25%) compared with group A (6.7%). Yet, the incidence of biopsy-proven acute rejection requiring steroid therapy was 6.7% in both groups. Hispanic race was common in groups A and B (87% and 74%). A total of six biopsies were performed in group B, with three patients having mild rejection responding to an increase in tacrolimus without the need for corticosteroids. One patient in group B was switched to cyclosporine for severe neurotoxicity and remains on monotherapy with normal graft function. No patient in either group developed a requirement for additional antihypertensive medication. Likewise, there were no patients with new-onset diabetes. The bone mineral densitometry was higher in group B at every time point but did not reach statistical significance. Serum cholesterol level was significantly (P=0.03) lower in group B at 6 months after OLTx. Serum triglycerides were also lower, but the difference was not significant. Quantitative polymerase chain reaction for HCV-positive patients (group A, n=7; group B, n=8) frequently increased after OLTx. There was no correlative decrease associated with daclizumab. At present, two patients in group A have documented HCV recurrence. CONCLUSION: Corticosteroids can be safely avoided after POD 1 with the current regimen. With early follow-up, there is no difference in hypertension or diabetes or bone density. Lipid panels tended to be lower in patients who were not on corticosteroids. Longer term follow-up will be needed to demonstrate the potential advantage of corticosteroid avoidance in regard to hypertension, diabetes, and possibly HCV recurrence. PMID- 11726832 TI - Comparison of hepatic artery and portal vein reperfusion during orthotopic liver transplantation. AB - BACKGROUND: During orthotopic liver transplantation (OLT), it is standard procedure to reperfuse the liver via the portal vein (PV) despite having a lower oxygen content and perfusion pressure than the hepatic artery (HA). There are no published studies that describe graft function and outcome when the HA is used for reperfusion. We report a retrospective comparison of graft outcome after HA or PV reperfusion when the piggyback technique was used. METHODS: We identified 26 patients who had undergone OLT with HA reperfusion and 26 patients reperfused via the PV. Demographics, primary diagnosis, surgeon, warm and cold ischemic times, and blood product use were recorded. In each patient, whole blood lactate concentration, prothrombin time (PT), and alanine aminotransferase (ALT) were measured at defined time points during and after surgery as indices of graft lactate metabolism, synthetic function, and reperfusion injury, respectively. Thirty-day and 1-year outcome data were recorded. Data were compared between the HA and PV groups. RESULTS: Demographics, blood product use, primary diagnosis, cold ischemic time, and surgeon were similar between the groups. Warm ischemic time was longer in the HA group (mean [SD] HA 51.2 [14.7], PV 40 [9.1] min, P=0.002). Blood lactate concentrations were similar at all time points. There was no difference in 24-hr postoperative PT between the groups (median [InterQuartile (IQ) range] HA 17.5 [16-28.3], PV 19 [16-24] sec, P=0.85). Peak postoperative ALT values were comparable (median [IQ range] HA 1031 [668-1701], PV 1107 [754-1824] IU/ml, P=0.78). There were no statistically significant differences in 30-day or 1-year mortality, but more early deaths occurred in the HA group. Using our data, we calculated that a prospective randomized trial would need approximately 300 patients to be sure that mortality was the same with both techniques. CONCLUSION: We have demonstrated no clinically or statistically significant differences in indices of graft function, reperfusion injury, or outcome between primary HA or PV reperfusion. PMID- 11726833 TI - Liver transplantation in adults coinfected with HIV. AB - OBJECTIVE: To report our experience of prospectively identifying and transplanting livers into HIV-positive patients. DESIGN: Liver transplantation in HIV-positive patients remains controversial. The finding of HIV is usually considered a contraindication to any form of transplantation. Previously reported cases are few and refer to patients who tested HIV positive after they had their liver transplantations or who seroconverted in the posttransplantation period. This is, to our knowledge, the only report of patients who were known to be HIV positive at the time of decision for listing for transplantation. METHODS: The medical records of five HIV-positive patients who received liver transplants in King's College Hospital, London, during a 5-year period (January 1995-December 1999) were reviewed. All five were known to be HIV positive at the time of listing for liver replacement. Three of them had end-stage liver disease due to hepatitis C (two of them had underlying Hemophilia A) while the other two had acute liver failure, one due to hepatitis B infection and one due to nonA-nonB nonC hepatitis. In all but one patient the HIV infection had been asymptomatic. RESULTS: All patients survived the immediate posttransplantation period, but the three patients with hepatitis C died of complications of recurrent hepatitis C between 6 and 25 months posttransplantation. The other two patients are currently alive 4 and 34 months posttransplantation with good graft function and without complications from their HIV infection. CONCLUSION: The early outcome of liver transplantation in HIV seropositive patients can be good, and patients should not be excluded from transplantation if their liver disease determines their prognosis. More effective antiviral therapy for hepatitis C given posttransplantation, and for hepatitis B reinfection, should improve the longer term outcome of HIV patients with end-stage liver disease due to hepatitis. PMID- 11726834 TI - Hyperuricemia, gout, and renal function after liver transplantation. AB - BACKGROUND: Hyperuricemia is a recognized complication of renal and cardiac transplantation, but the development of hyperuricemia and gout following liver transplantation have received less attention. We have retrospectively assessed the prevalence of hyperuricemia in 134 consecutive liver transplant recipients. RESULTS: Forty-seven percent of the liver transplant recipients studied had hyperuricemia. Serum creatinine was higher in hyperuricemic than in nonhyperuricemic patients. Peak uric acid correlated significantly with corresponding serum creatinine (rs=0.694). Only 6% developed gout. All the patients with gout and 10 hyperuricemic patients with renal impairment but without gout were treated with allopurinol. Over a median period of 3 months, mean serum creatinine fell from 177 micromol/l to 160 micromol/l (P=0.01), without change in type or dose of immuno-suppression. CONCLUSIONS: There is an important association between liver transplantation and hyperuricemia. Treatment with allopurinol results in a significant reduction in serum creatinine in patients with gout and in those with hyperuricemia and renal impairment. PMID- 11726835 TI - Microcirculatory failure after rat liver transplantation is related to Kupffer cell-derived oxidant stress but not involved in early graft dysfunction. AB - BACKGROUND: Microcirculatory failure, activation of Kupffer cells (KC), and the formation of reactive oxygen species (ROS) are considered pivotal mechanisms of reperfusion injury after orthotopic liver transplantation. However, the sequence of these events and their impact on early graft function remain controversial. We therefore investigated whether KC induce microcirculatory disturbances through ROS release and whether microcirculatory failure contributes to early graft function after liver transplantation. METHODS: Donor livers of Lewis rats were pretreated either with saline or with gadolinium chloride (GdCl3), an inhibitor of KC function (n=8 each). Syngeneic OLT was performed after 24 hr of hypothermic preservation in University of Wisconsin solution. RESULTS: Intravital microscopy revealed significantly higher sinusoidal perfusion rates in GdCl3-treated allografts (92+/-1.1% vs. 75.7+/-0.8%; P<0.001) compared with untreated controls; permanent leukocyte sticking in sinusoids (23.5+/-2.1 vs. 62.6+/-3.3 cells/lobule, P<0.001) and in postsinusoidal venules (153.1+/-10.4 vs. 446.6+/ 46.4 cells/mm(2), P<0.001) were markedly attenuated in GdCl3-treated allografts. Improvement of microcirculatory parameters in GdCl3-treated livers was correlated with a significant reduction of plasma glutathione disulfide formation by KC derived ROS (0.96+/-0.1 microM vs. 1.79+/-0.5 microM; P<0.01). Despite these beneficial effects, GdCl3-pretreatment failed to improve postischemic alanine aminotransferase release and bile flow. CONCLUSIONS: Microcirculatory failure after liver transplantation is related to KC-derived oxidant stress but not involved in early graft dysfunction. PMID- 11726836 TI - Effects of cyclosporine and hydrocortisone on Kaposi's sarcoma-associated herpesvirus genome replication and cell apoptosis induction. AB - BACKGROUND: Iatrogenic immunosuppressed patients are at increased risk for development of various cancers that comprise Kaposi's sarcoma (KS). METHODS: To investigate the direct impact of immunosuppressive agents on Kaposi's sarcoma associated herpesvirus (KSHV) and KS development, we quantified the effects of cyclosporine (CsA) and hydrocortisone (HC) on KSHV genome replication and the consequences on the cell survival. RESULTS: In the presence of phorbol ester 12-O tetradecanoyl phorbol-13-acetate, we observed an increase of intracellular and extracellular KSHV DNA concomitantly with an increase of gp (glycoprotein) K8.1 expression, indicating KSHV genome replication. This replication was accompanied by cell apoptosis. In comparison, in the presence of CsA, HC, or both, we did not observe any effect on KSHV replication or gp K8.1 expression. CONCLUSION: Our results suggest that immunosuppressive agents such as HC and CsA do not activate the lytic cycle of KSHV and do not modify the cell survival thus promoting cancer progression by a direct cellular effect. PMID- 11726837 TI - Tacrolimus for the treatment of gout in renal transplantation: two case reports and review of the literature. AB - Episodes of gout are common in the setting of renal transplantation. Hyperuricemia and gout have been associated with the use of the calcineurin inhibitor, cyclosporine. We report two cases of severe polyarticular gout resistant to conventional therapy in renal transplant recipients that resolved after switching from cyclosporine to tacrolimus-based immunosuppression. There was no alteration in renal function, and trough concentrations of both cyclosporine and tacrolimus were within the recommended range. Resolution of gout occurred within a month of discontinuation of cyclosporine and commencement of tacrolimus. Use of tacrolimus may be beneficial in the renal transplant recipient with refractory gout. PMID- 11726838 TI - Intragraft activation of genes encoding cytotoxic T lymphocyte effector molecules precedes the histological evidence of rejection in human cardiac transplantation. AB - BACKGROUND: The purpose of the present study was to investigate transcripts of perforin, granzyme B, and Fas ligand (FasL) in heart transplants undergoing rejection. METHODS: Quantitative reverse transcriptase-polymerase chain reaction was applied for mRNA detection in 29 endomyocardial biopsy specimens from 11 cardiac allograft recipients. RESULTS: The mRNA levels of granzyme B, perforin, and FasL were higher (P<0.05) in biopsy specimens with rejection than in biopsy specimens without rejection (granzyme B, 0.53 vs. 0.09; perforin, 0.34 vs. 0; FasL, 0.57 vs. 0.36). In prerejection biopsy specimens, granzyme B and FasL levels were significantly higher than in biopsy specimens without rejection. Any two of the three transcripts were increased in 100% of prerejection, in 92% of rejection, and in 36% of no rejection biopsy specimens (P<0.04). CONCLUSIONS: The assessment of intragraft levels of cytotoxic T lymphocyte effector molecule mRNA represents a valuable tool in the monitoring of cardiac allograft rejection, especially considering the predictive value for warning of impending acute rejection. PMID- 11726839 TI - Use of leflunomide in human renal transplantation. PMID- 11726840 TI - Two doses of daclizumab are sufficient for prolonged interleukin-2Ralpha chain blockade. PMID- 11726841 TI - Endoscopic injection therapy in bleeding Mallory-Weiss syndrome: a randomized controlled trial. AB - BACKGROUND: Endoscopic injection is widely used in the therapy of bleeding gastroduodenal ulcers, but its role in the management of bleeding Mallory-Weiss tears has not been properly assessed. METHODS: Sixty-three patients undergoing emergency endoscopy in whom there was a high index of suspicion that a Mallory Weiss tear was the source of bleeding were randomly assigned to undergo endoscopic injection therapy (epinephrine and polidocanol) or no endoscopic therapy in 2 university-affiliated hospitals. Rates of recurrent bleeding, transfusion requirements, complications, mortality, and length of hospital stay were determined for both groups of patients. RESULTS: Bleeding recurred in 8 patients in the control group versus only 2 in the endoscopic treatment group (25.8% vs. 6.2%, p < 0.05). Hospital stay was longer for the control group (5.5 +/- 0.2, median 6.0, range 2.0-8.0 days vs. 3.4 +/- 0.2, median 3.0, range 2.0 6.0 days; p < 0.001). There was a trend toward a higher transfusion requirement after endoscopy in the control group versus the patients treated by injection (0.9 +/- 0.2, median 0.0, range 0.0-4.0 units vs. 0.2 +/- 0.1, median 0.0, range 0.0-2.0 units; p = 0.09). No complications or adverse events caused by endoscopic injection were noted. Two patients in the control group died of causes unrelated to bleeding. CONCLUSIONS: Endoscopic injection therapy is a useful option in the management of patients with Mallory-Weiss syndrome at high risk for recurrent bleeding. PMID- 11726842 TI - Combined endoscopic mucosal resection and photodynamic therapy for esophageal neoplasia within Barrett's esophagus. AB - BACKGROUND: Endoscopic mucosal resection (EMR) and photodynamic therapy have been proposed as treatments for early stage cancers. EMR is limited by its focal nature whereas photodynamic therapy is dependent on precise staging. The combination of EMR and photodynamic therapy were studied in the treatment of superficial cancer in patients with Barrett's esophagus. METHODS: Seventeen consecutive nonsurgical patients with superficial cancers underwent EMR followed by photodynamic therapy with a porphyrin photosensitizer. Photoradiation was performed at 630 nm for a total dose of 200 J/cm of diffuser. RESULTS: Seventeen patients (15 men; mean age 69 +/- 13 years) underwent EMR. The mean diameter of mucosal resection was 1 cm. The margins were involved by cancer in 3 cases. EMR improved staging in 8 patients (47%). Sixteen (94%) patients remained in remission (median follow-up 13 months). Complications included minor bleeding after EMR in 1 patient (6%), stricture in 5 (30%), cutaneous phototoxicity in 2 (12%), and supraventricular tachycardia in 1 patient (6%). CONCLUSIONS: Combined EMR and photodynamic therapy appears to be an effective and safe therapy for superficial esophageal cancer within Barrett's esophagus. This combination improves cancer staging, removes the superficial cancer, and eliminates remaining mucosa at risk for cancer development. PMID- 11726843 TI - Accuracy of EUS in the evaluation of Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma. AB - BACKGROUND: Nonoperative therapy with intent to cure may be considered for patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma. However, a more advanced stage of disease must be precluded before such treatment. The potential of EUS for this purpose was evaluated. METHODS: EUS was performed in patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma based on endoscopy, endoscopic biopsies, and CT before esophagectomy. EUS findings were compared with surgical/pathologic evaluation. RESULTS: EUS suggested submucosal invasion in 6 patients and lymph node involvement in 5 patients. By surgical/pathologic evaluation, 5 of 22 patients (23%) had unsuspected submucosal invasion and 1 had lymph node involvement. EUS detected all 5 instances of submucosal invasion and the single instance of lymph node involvement. EUS was falsely positive for submucosal invasion in 1 patient and for lymph node involvement in 4 patients. Sensitivity, specificity, and negative predictive values of preoperative EUS for submucosal invasion were 100%, 94%, and 100%, and for lymph node involvement were 100%, 81%, and 100%, respectively. A nodule or stricture noted by endoscopy was associated with an increased likelihood of submucosal invasion. CONCLUSIONS: In patients with Barrett's esophagus and high-grade dysplasia or intramucosal carcinoma, EUS detected otherwise unsuspected submucosal invasion and lymph node involvement. Patients should be evaluated with EUS when nonoperative therapy is contemplated. PMID- 11726844 TI - Comparison of sphincter of Oddi manometry, fatty meal sonography, and hepatobiliary scintigraphy in the diagnosis of sphincter of Oddi dysfunction. AB - BACKGROUND: Sphincter of Oddi dysfunction (SOD) afflicts approximately 1% to 5% of patients after cholecystectomy. The diagnostic standard for SOD is sphincter of Oddi manometry (SOM), a technically difficult, invasive test that is frequently complicated by pancreatitis. A sensitive and accurate noninvasive imaging modality is thus needed for the diagnosis of SOD. Quantitative hepatobiliary scintigraphy (HBS) and fatty meal sonography (EMS) are frequently used for this purpose, but results vary. This study compared SOM, HBS, and EMS in the diagnosis of SOD in a large group of patients. METHODS: Three hundred four consecutive patients after cholecystectomy (38 men, 266 women, age 17-72 years) suspected to have SOD were evaluated by SOM, FMS, and HBS. SOM was considered abnormal if any of the following were observed: (1) increased basal pressure (greater than 40 mm Hg), (2) increased phasic activity with amplitude greater than 350 mm Hg, (3) frequency of contractions greater than 8 per minute, (4) greater than 50% of propagation sequences retrograde, and (5) paradoxical response to cholecystokinin. FMS was considered abnormal if ductal dilation was greater than 2 mm at 45 minutes after fatty meal ingestion. Quantitative HBS was performed with sequential images obtained every 5 minutes for 90 minutes to monitor excretion of the radionuclide. Time-to-peak, halftime, and downslope were calculated according to predetermined ranges. RESULTS: A diagnosis of SOD was made in 73 patients (24%) by using SOM as the reference standard. HBS was abnormal in 86 whereas EMS was abnormal in 22 patients. A true-positive result was obtained in 15 patients by EMS and 36 patients with HBS. EMS and HBS gave false-positive results, respectively, in 7 and 50 patients. Sensitivity of EMS was 21% and for HBS 49%, whereas specificities were 97% and 78%, respectively. EMS, HBS, or both were abnormal in 90% of patients with Geenen-Hogan Type I SOD, 50% with Type II, and 44% of Type III. Of the 73 patients who underwent sphincterotomy, 40 had a long-term response. Of those with SOD, 11 of 13 patients (85%) with an abnormal HBS and EMS had a good long-term response. CONCLUSIONS: In this series, the largest reported to date, correlation of FMS and HBS with SOM in the diagnosis of SOD was poor. When HBS and EMS are used together, a slight increase in sensitivity can be expected. The accuracy of EMS and HBS in the diagnosis of SOD decreases across the spectrum from Type I to Type III SOD. EMS and HBS, nonetheless, may by of assistance in predicting long-term response to endoscopic sphincterotomy in patients with elevated sphincter of Oddi basal pressure. PMID- 11726845 TI - Efficacy and safety of sodium phosphate tablets compared with PEG solution in colon cleansing: two identically designed, randomized, controlled, parallel group, multicenter phase III trials. AB - BACKGROUND: Liquid purgatives for cleansing before colonoscopy often are poorly tolerated. A sodium phosphate tablet has been developed to provide equivalent efficacy with better patient tolerance. These 2 studies compare the safety, efficacy, and patient acceptance of the tablet (Visicol) to a polyethylene glycol (PEG) solution in adults undergoing colonoscopy. METHODS: Two identically designed, randomized, investigator-blinded, multicenter trials were performed. The primary efficacy variable was the overall quality of colon cleansing. Patient tolerance was assessed in terms of compliance with the dosing regimen. Safety assessments included recording of adverse events and changes in biochemical tests, electrocardiogram, and vital signs. RESULTS: Eight hundred forty-five patients participated in the studies; 420 took sodium phosphate tablets and 425 took a PEG solution. The 2 methods of preparation were equivalent in the overall quality of colon cleansing, cleansing in the right colon, and the frequency of inadequate preparation. Overall cleansing was excellent or good in 84.3% of patients in the tablet group and in 76.7% in the PEG group. Patient compliance was greater in the tablet group. There were also significantly fewer GI side effects in this group. CONCLUSIONS: Sodium phosphate tablets, compared with PEG solution, produce equivalent colon cleansing, are associated with fewer GI side effects, and are better tolerated by patients. PMID- 11726846 TI - The utility of EUS and EUS-guided fine needle aspiration in detecting celiac lymph node metastasis in patients with esophageal cancer: a single-center experience. AB - BACKGROUND: The aims of this study were to determine the utility of EUS and EUS guided fine needle aspiration (EUS-FNA) in the detection and confirmation of celiac lymph node metastasis in patients with esophageal cancer and to define EUS features predictive of celiac lymph node metastasis in these patients. METHODS: The records of 211 patients with esophageal cancer who underwent EUS staging were reviewed. The operating characteristics of EUS were determined in patients where either surgery, EUS-FNA of a celiac lymph node, or both were performed (n = 102). The association between selected variables and the presence of celiac lymph node metastasis was evaluated by univariate and multivariable analyses. RESULTS: EUS in 48 patients provided a true-positive diagnosis of celiac lymph node involvement, a false-positive and false-negative result, respectively, in 6 and 14 patients, and a true-negative diagnosis in 34 patients. The sensitivity of EUS in detecting celiac lymph node was 77% (95% CI [67, 88]), specificity 85% (95% CI [74, 96]), negative predictive value 71% (95% CI [58, 84]), and the positive predictive value 89% (95% CI [81, 97]). EUS-FNA was performed in 94% (51/54) of patients with celiac lymph nodes. The accuracy of EUS-FNA in detecting malignant celiac lymph nodes was 98% (95% CI [90, 100]). Advanced T-stage, the need for dilation, detection of peritumoral lymph nodes, and black race were associated with celiac lymph node involvement. In multivariable analysis, advanced T-stage was the strongest predictor of celiac lymph node involvement. CONCLUSION: EUS and EUS-FNA are highly accurate in detecting and confirming celiac lymph nodes metastasis. Depth of tumor invasion as assessed by EUS is a strong predictor of celiac lymph node metastasis in patients with esophageal cancer. PMID- 11726847 TI - Over-the-wire US catheter probe as an adjunct to ERCP in the detection of choledocholithiasis. AB - BACKGROUND: Intraductal ultrasound (IDUS) as an adjunct to ERCP for detection of extrahepatic bile duct stones is technically easy, accurate, and safe. This prospective study evaluated IDUS with an "over-the-wire" catheter US probe as an adjunct to ERCP. METHODS: Sixty-five patients, highly suspected to have choledocholithiasis, underwent IDUS during ERCP. The IDUS probe was inserted by means of the duodenoscope into the bile duct without performing a sphincterotomy. All stones identified by IDUS or retrograde cholangiography were removed with either a basket or retrieval balloon after endoscopic sphincterotomy. RESULTS: The final diagnosis was choledocholithiasis in 59 patients. Bile duct diameter ranged from 0.6 to 2.3 cm and stone size from 2 mm to 2 cm. IDUS successfully identified all stones in these patients. IDUS resulted in 2 false-positive diagnoses in the remaining 6 patients without stones (overall accuracy 97%, sensitivity 100%, specificity 67%). Cholangiography detected stones in 55 of the patients with stones (accuracy 94%, sensitivity 93%, specificity 100%). CONCLUSION: IDUS, a safe, technically easy procedure, is highly accurate in the detection of extrahepatic bile duct stones regardless of the diameter of the bile ducts. The "over-the-wire" technique preserves access to the cannulated duct. IDUS is an excellent adjunct to ERCP for the diagnosis of choledocholithiasis. IDUS differentiates stones from air bubbles and prevents unnecessary sphincterotomy. PMID- 11726848 TI - Early EUS of the bile duct before endoscopic sphincterotomy for acute biliary pancreatitis. AB - BACKGROUND: Endoscopic sphincterotomy can benefit patients with suspected biliary pancreatitis, although there are procedure-related complications. EUS can be used to select patients for endoscopic sphincterotomy. The results of this strategy were assessed. METHODS: Information on patients referred for EUS were recorded in a database. One hundred twenty-three patients with suspected biliary pancreatitis (57 men, 66 women; median age 55 years) were included and followed. All underwent EUS followed by endoscopic sphincterotomy during the same procedure if choledocholithiasis was identified. Outcomes were studied in relation to the initial severity of biliary pancreatitis (Ranson and Balthazar scores), presence of stones, and time span between onset of biliary pancreatitis and EUS plus endoscopic sphincterotomy. RESULTS: Thirty-five patients (28%) had a Ranson score greater than 3 on admission and 38 (31%) were Balthazar D-E. The median time from admission to EUS was 3 days. EUS imaging of the bile duct was complete in all but 3 patients. Thirty-three patients (27%) had choledocholithiasis on EUS and underwent endoscopic sphincterotomy. Stones were more frequent in patients with jaundice (p < 0.005) and when EUS was performed less than 3 days after admission (p < 0.05). One hundred patients (81%) recovered without complication. Two patients (1.6%) died, 1 had recurrent BP develop, 6 (5%) had further biliary symptoms, and 16 (13%) had complications of pancreatitis develop (9 pseudocysts). There were 3 mild endoscopic sphincterotomy-related complications (complication rate 6.5%). CONCLUSIONS: In this series in which endoscopic sphincterotomy was performed selectively depending on the endosonographic presence or absence of ductal stones early in the course of the pancreatitis, and not according to its predicted severity, mortality and complications of endoscopic sphincterotomy were low and unrelated to the predicted severity of biliary pancreatitis or the presence of choledocholithiasis. Controlled trials are needed to confirm the superiority of this strategy compared with ERCP alone for the management of biliary pancreatitis. PMID- 11726849 TI - Pseudomembranous esophagitis. AB - BACKGROUND: Little is known of pseudomembranous esophagitis, a condition of striking endoscopic appearance. Presented here is a description of its nature and outcome. METHOD: Information on all patients with upper GI disease seen in our unit are held in a computerized database that includes presentation, diagnosis (including pseudomembranous esophagitis), treatment, and outcome. RESULTS: Forty eight patients (mean age 70 years) with pseudomembranous esophagitis were seen over 15 years; 42 were in-patients with various illnesses. At endoscopy, a thin, concentric membrane and/or thick slough covered the distal half and occasionally the entire esophagus. This layer, yellow or blackened, could be peeled away to reveal underlying friable esophageal submucosa. The membrane was composed of fibrinous exudate and inflammatory cells; there was no basement membrane, hence the term pseudomembrane. Patients were treated with histamine H2 receptor antagonists or proton pump inhibitors for about 3 months and followed by endoscopy or clinical observation. All became asymptomatic; the pseudomembrane had disappeared in 32 who underwent follow-up endoscopy. It recurred in 3 of 38 being followed (mean 3.2 years), again associated with another illness requiring hospitalization. The long-term outcome was poor, determined by age and general condition and independent of pseudomembranous esophagitis. Seven patients died within 3 months and 17 during follow-up (mean 42 months, range 5-140 months). CONCLUSION: Pseudomembranous esophagitis is an unusual condition of unknown cause, probably under-reported and associated with systemic illness. It heals rapidly and recurrence is uncommon. PMID- 11726850 TI - Endoscopic features of celiac disease in children. AB - BACKGROUND: Endoscopic abnormalities have been described in adult patients with celiac disease that may suggest the diagnosis, especially when the presentation is atypical. METHODS: The duodenum of 140 children undergoing EGD for various different indications was evaluated macroscopically and histologically. RESULTS: Histology revealed total villous atrophy in 80 patients, 79 of whom had celiac disease. Among these, 100% had a mucosal mosaic pattern in the duodenum (sensitivity 98.7%, specificity 96.7%, positive predictive value 97.5%, negative predictive value 98.3%), 70% had scalloped duodenal folds (sensitivity 68.7%, specificity 98.3%, positive predictive value 98.2%, negative predictive value 70.2%), 15% had visible vasculature, and 6% had reduction of duodenal folds. Sensitivity and specificity of endoscopic findings were not modified by chromoendoscopy. Except for the mosaic pattern, the frequency of endoscopic abnormalities increased with age; reduction of duodenal folds was never seen in children with celiac disease who were less than 5 years of age. CONCLUSIONS: The frequency and diagnostic value of endoscopic abnormalities are different in children with celiac disease compared with adults with this disease. Because indications for endoscopy, such as abdominal pain, dyspepsia, and unexplained anemia, can be manifestations of celiac disease, and villous atrophy may have a patchy distribution, awareness of these endoscopic abnormalities is important in the diagnosis of celiac disease in children. PMID- 11726851 TI - Endoscopy and ERCP in the setting of previous upper GI tract surgery. Part I: reconstruction without alteration of pancreaticobiliary anatomy. PMID- 11726852 TI - Colonic mucosal pseudolipomatosis. PMID- 11726853 TI - Histologic findings after cyanoacrylate injection. PMID- 11726854 TI - Endoscopic images in cricopharyngeal dysfunction. PMID- 11726855 TI - Duodenal plasmacytoma. PMID- 11726856 TI - Treatment of symptomatic diffuse esophageal spasm by endoscopic injections of botulinum toxin: a prospective study with long-term follow-up. AB - BACKGROUND: Diffuse esophageal spasm is a rare esophageal motility disorder for which there are no satisfactory pharmacologic alternatives for treatment. The aim of this study was to investigate whether botulinum toxin (BTX) injection is an effective short- and long-term treatment for patients with symptoms caused by diffuse esophageal spasm. Whether recurrence of clinical symptoms can be successfully retreated by BTX injection was also studied. METHODS: Nine symptomatic patients (6 women, 3 men; 57-86 years) with manometrically proven diffuse esophageal spasm underwent BTX injection. One hundred IU BTX were diluted in l0 mL of saline solution and injected endoscopically at multiple sites along the esophageal wall beginning in the region of the lower esophageal sphincter and moving proximally in 1- to 1.5-cm intervals, and into endoscopically visible contraction rings. Symptom scores based on an analogue scale for dysphagia, regurgitation, and noncardiac chest pain were assessed before and after therapy, 1 day thereafter, and at 1 and 6 months. RESULTS: Symptoms improved immediately in 7 (78%) patients after 1 injection session. After 4 weeks 8 (89%) patients were in remission with a decrease in total symptom score. The total symptom score decreased from a median 8.0 (interquartile range: 6.75; 9.0) before treatment to 2.0 (1.5; 3.75) after 1 day (p < 0.01) and to 2.0 (interquartile range: 0.75; 3.0) after 1 month (p < 0.01). After 6 months all 8 patients with a response at 1 month still had a symptom score of 3 or less without further treatment. Subsequently 4 patients required reinjection 8, 12, 15, or 24 months after the initial treatment with similarly good results. No serious adverse effects were observed. CONCLUSIONS: BTX injection at several levels of the tubular esophagus is an effective treatment for patients with symptoms caused by diffuse esophageal spasm. Symptom relapse can be effectively treated by repeated BTX injection. PMID- 11726857 TI - Endoscopic treatment of Zenker's diverticulum with an oblique-end hood attached to the endoscope. AB - BACKGROUND: The feasibility and efficacy of incision of the septum between the pharyngoesophageal (Zenker's) diverticulum and the esophagus with a flexible endoscope and needle-knife have been demonstrated. Exposure of the septum remains a critical element of the procedure. This is a report of a technical improvement in this technique: use of a transparent oblique-end hood attached to the tip of the endoscope. METHODS: Ten patients (10 men, age range 67-87 years) with Zenker's diverticulum were treated. With the aid of an oblique-end hood attached to the tip of the endoscope, the septum between the Zenker's diverticulum and esophagus was incised with pure coagulation current. A 10F nasogastric tube was left in place for 48 hours after the procedure to provide nutrition. Thereafter, oral intake was progressively resumed. RESULTS: Complete incision of the septum was achieved in a single session in all cases. Bleeding or perforation did not occur in this small series. Complete relief of dysphagia was reported by all patients during follow-up that ranged from 2 to 12 months. CONCLUSIONS: With the use of the oblique-end hood attached to the tip of the endoscope, incision of Zenker's diverticulum is simplified and can be performed as a single-session procedure. PMID- 11726858 TI - Photodynamic therapy with 5-aminolevulinic acid is not effective in bile duct cancer. AB - BACKGROUND: The results of preliminary studies of photodynamic therapy (PDT) for palliation of patients with bile duct cancer with hematoporphyrin derivative have been good. Rapid elimination of a photosensitizer could potentially shorten the time requirement for shielding from light. This would enhance the benefit of this form of palliative treatment. Therefore the feasibility of PDT was investigated for nonresectable bile duct cancer by using 5-aminolevulinic acid. METHODS: Four patients with nonresectable bile duct cancer underwent cholangiography, cholangioscopy, and intraductal US before PDT. Light activation was performed 5 to 7 hours after oral administration of 5-aminolevulinic acid. All patients had an endoprosthesis placed in the bile duct after PDT. RESULTS: Cholangioscopy 72 hours after PDT revealed superficial fibrinoid necrosis. However, 4 weeks after PDT there was no significant reduction in bile duct stenoses. Two patients had infectious complications develop, but phototoxicity was not observed. CONCLUSIONS: Although superficial tumor necrosis was evident, PDT with 5 aminolevulinic acid failed to significantly reduce malignant bile duct obstruction. Therefore 5-aminolevulinic acid-PDT cannot be recommended for the palliative treatment of bile duct cancer. PMID- 11726859 TI - Primary cricopharyngeal dysfunction: treatment with balloon catheter dilatation. AB - BACKGROUND: Primary cricopharyngeal dysfunction (PCD) is a rare, idiopathic, functional disorder of the upper esophageal sphincter, characterized by dysphagia, frequent aspiration, and functional narrowing at the level of the upper esophageal sphincter. METHODS: Five of 29 patients with oropharyngeal dysphagia were found to have PCD. Patients presented with severe dysphagia and predisposition to aspiration. Radiography demonstrated narrowing at the level of the upper esophageal sphincter and aspiration. An endoscope could be introduced into the esophagus in only 2 patients before dilation. OBSERVATIONS: In contrast to organic stenoses, these functional upper esophageal sphincter stenoses were dilated without difficulty with a balloon catheter. After low-pressure (1.5-2 atm) progressive balloon dilation (to 20 mm), superficial mucosal injury was observed only in one patient. After dilatation, symptoms resolved and barium swallow demonstrated normal passage through the upper esophageal sphincter. During a mean follow-up of 21 months (7-33), redilation was necessary in only 1 case. CONCLUSIONS: Balloon catheter dilatation of PCD is minimally invasive and provides both important diagnostic information and effective therapy. It should be the first choice of therapy for PCD. PMID- 11726860 TI - Duodenal mucosa-associated lymphoid tissue lymphoma treated by eradication of Helicobacter pylori: report of 2 cases including EUS findings. PMID- 11726861 TI - An extraordinary Dieulafoy's lesion presenting as varices of the gastric fundus. PMID- 11726862 TI - Endoscopic treatment of massive lower GI hemorrhage in two patients with ulcerative colitis. PMID- 11726863 TI - Cap-assisted endoscopic submucosal resection of a pancreatic rest. PMID- 11726864 TI - Successful treatment of a bleeding esophageal sclerotherapy ulcer with endoscopic injection of granulocyte-macrophage colony-stimulating factor. PMID- 11726865 TI - Rectal varices successfully treated by endoscopic injection sclerotherapy after careful hemodynamic evaluation: a case report. PMID- 11726866 TI - Acute pancreatitis caused by a prolapsing gastrostomy tube. PMID- 11726867 TI - EUS: a role in metastatic cancer with undiagnosed primary? PMID- 11726868 TI - An occupational GI cancer screening program. PMID- 11726869 TI - Topical pharyngeal anesthesia during gastroscopy. PMID- 11726870 TI - Can unilateral stent placement be adequate in patients with Klatskin tumors of Bismuth-type III and IV? PMID- 11726871 TI - Laparoscopically assisted panenteroscopy. PMID- 11726872 TI - The modeling of relationships among variables with regression techniques. PMID- 11726873 TI - Guidelines for credentialing and granting privileges for endoscopic ultrasound. PMID- 11726874 TI - The role of endoscopy in dyspepsia. PMID- 11726875 TI - Diagnostic laparoscopy guidelines for clinical application. PMID- 11726876 TI - Use of endoscopy in diarrheal illnesses. PMID- 11726877 TI - Technology status evaluation report. Transmission of infection by gastrointestinal endoscopy. May 2001. PMID- 11726878 TI - Technology Status Evaluation report. Colonoscopy preparations. May 2001. PMID- 11726879 TI - Technology status evaluation report. Endoscopic hemostatic devices. May 2001. PMID- 11726880 TI - Ischemic mitral regurgitation redux--to repair or to replace? PMID- 11726881 TI - Clinical-pathologic conference: use and choice of statistical methods for the clinical study, "superficial adenocarcinoma of the esophagus". PMID- 11726882 TI - Superficial adenocarcinoma of the esophagus. AB - OBJECTIVE: Experience with treatment and outcome of superficial adenocarcinoma of the esophagus is limited. The purpose of this study was to evaluate the results of surgical management and identify predictors of survival. METHODS: Between September 1985 and December 1999, 122 patients underwent resection. Eighty-nine percent were men (mean age 63 +/- 10 years; range 35-83 years). Sixty (49%) patients were in endoscopic surveillance programs and 48 (39%) had the preoperative diagnosis of high-grade dysplasia. Forced expiratory volume in 1 second was less than 2 L in 12 (12%). Seventy-five (61%) patients underwent transhiatal esophagectomy. Pathologic stage was N1 in 8 (7%). Pulmonary complications necessitating reintubation (respiratory failure) occurred in 10 (8%) patients. Time-related survival models were developed for decision-making (preoperative), prognosis (operative), and hospital care (postoperative). RESULTS: Operative mortality was 2.5%. Survival at 1, 5, and 10 years was 89%, 77%, and 68%. Preoperative decision-making factors associated with ideal outcome were 1-second forced expiratory volume of more than 2 L, surveillance, preoperative diagnosis of high-grade dysplasia, and planned transhiatal esophagectomy. Prognosis was decreased in younger patients and in those with N1 disease. Postoperative respiratory failure increased mortality. CONCLUSIONS: Surgery is the treatment of choice for superficial adenocarcinoma of the esophagus. The ideal patient has a preoperative diagnosis of high-grade dysplasia found at surveillance, good pulmonary function, and undergoes a transhiatal esophagectomy. Discovery of N1 disease or development of postoperative respiratory failure reduces the benefits of surgery. PMID- 11726883 TI - Lobar torsion after pulmonary resection: presentation and outcome. AB - OBJECTIVE: We reviewed our experience on postoperative lobar torsion. METHODS: Between January 1972 and January 1998, 7887 patients underwent pulmonary resection at our institution. Seven (0.089%; 4 women and 3 men; median age, 68 years) patients required surgical reintervention for lobar torsion. RESULTS: The indications for pulmonary resection were non-small cell carcinoma in 5 patients, lymphoma in 1 patient, and metastatic prostate carcinoma in 1 patient. The right upper lobe was resected in 3 patients, the left lower lobe in 2 patients, and the right middle and right lower lobe in 1 patient each. Postoperative radiographs demonstrated pulmonary infiltrates and volume loss in 5 patients and complete opacification in 2 patients. The median white blood cell count was 10.6 x 10(9) cells/L (range, 9.3-14.9 x 10(9) cells/L), and the median peak temperature was 38.4 degrees C (range, 37.8 degrees C-40.2 degrees C) during the first 48 hours postoperatively. The diagnosis of lobar torsion was made a median of 10 days (range, 2-14 days) after the initial operation; 4 patients underwent completion pneumonectomy, and 3 had lobectomy. Median hospitalization was 24 days and ranged from 10 to 56 days. There were no postoperative deaths. Complications after reoperation included respiratory failure in 2 patients, atrial arrhythmia in 2 patients, and empyema, urinary tract infection, and a transient ischemic attack in 1 patient each. CONCLUSIONS: Lobar torsion represents a difficult diagnostic dilemma in the early postoperative period after pulmonary resection. A high index of suspicion is necessary to avoid a delay in treatment. Late diagnosis results in further pulmonary resection and prolonged hospitalization in the majority of cases. PMID- 11726884 TI - An artificial lung reduces pulmonary impedance and improves right ventricular efficiency in pulmonary hypertension. AB - OBJECTIVE: Artificial lungs may have a role in supporting patients with end-stage lung disease as a bridge or alternative to lung transplantation. This investigation was performed to determine the effect of an artificial lung, perfused by the right ventricle in parallel with the pulmonary circulation, on indices of right ventricular load in a model of pulmonary hypertension. METHODS: Seven adult male sheep were connected to a low-resistance membrane oxygenator through conduits anastomosed end to side to the pulmonary artery and left atrium. Banding of the distal pulmonary artery generated acute pulmonary hypertension. Data were obtained with and without flow through the device conduits. Outcome measures of right ventricular load included hemodynamic parameters, as well as analysis of impedance, power consumption, wave reflections, cardiac efficiency, and the tension-time index. RESULTS: The model of pulmonary hypertension increased all indices of right ventricular load and decreased ventricular efficiency. Allowing flow through the artificial lung significantly reduced mean pulmonary artery pressure, zero harmonic impedance, right ventricular power consumption, amplitude of reflected waves, and the tension-time index. Cardiac efficiency was significantly increased. CONCLUSIONS: An artificial lung perfused by the right ventricle and applied in parallel with the pulmonary circulation reduces ventricular load and improves cardiac efficiency in the setting of pulmonary hypertension. These data suggest that an artificial lung in this configuration may benefit patients with end-stage lung disease and pulmonary hypertension with right ventricular strain. PMID- 11726885 TI - Clinical significance of tissue blood flow during esophagectomy by laser Doppler flowmetry. AB - OBJECTIVES: We sought to investigate the effect of tissue blood flow on the incidence of anastomotic leakage during esophagectomy. METHODS: Reconstruction was done with a gastric tube, and all cases involved cervical anastomosis. Tissue blood flow of 43 patients was measured with a laser Doppler flowmeter before emplacement and after anastomosis during surgical intervention. The reconstruction route and tissue blood flow before emplacement and that after anastomosis were analyzed as possible factors influencing anastomotic leakage. RESULTS: Tissue blood flow after anastomosis with leakage was 9.1 +/- 2.0 mL/min per 100 g, and that without leakage was 13.7 +/- 2.9 mL/min per 100 g. Tissue blood flow with leakage was significantly lower than that without leakage (P <.01, unpaired t test). Twenty-one patients had tissue blood flow after anastomosis of greater than 13 mL/min per 100 g, and none of them had leakage, whereas 5 patients had blood flow of less than 10 mL/min per 100 g, and all 5 had leakage. CONCLUSION: Tissue blood flow can be an important and useful indicator of the presence of current anastomotic leakage. Low tissue blood flow after anastomosis may mediate for appropriate surgical or pharmacologic interventions to detect, localize, and counteract leakage. PMID- 11726886 TI - Ischemic mitral valve reconstruction and replacement: comparison of long-term survival and complications. AB - OBJECTIVE: This study reviews the 223 consecutive mitral valve operations for ischemic mitral insufficiency performed at New York University Medical Center between January 1976 and January 1996. The results for mitral valve reconstruction are compared with those for prosthetic mitral valve replacement. METHODS: From January 1976 to January 1996, 223 patients with ischemic mitral insufficiency underwent mitral valve reconstruction (n = 152) or prosthetic mitral valve replacement (n = 71). Coronary artery bypass grafting was performed in 89% of cases of mitral reconstruction and 80% of cases of prosthetic replacement. In the group undergoing reconstruction, 77% had valvuloplasty with a ring annuloplasty and 23% had valvuloplasty with suture annuloplasty. In the group undergoing prosthetic replacement, 82% of patients received bioprostheses and 18% received mechanical prostheses. RESULTS: Follow-up was 93% complete (median 14.6 mo, range 0-219 mo). Thirty-day mortality was 10% for mitral reconstruction and 20% for prosthetic replacement. The short-term mortality was higher among patients in New York Heart Association functional class IV than among those in classes I to III (odds ratio 5.75, confidence interval 1.25-26.5) and was reduced among patients with angina relative to those without angina (odds ratio 0.26, confidence interval 0.05-1.2). The 30-day death or complication rate was similarly elevated among patients in functional class IV (odds ratio 5.53; confidence interval 1.23-25.04). Patients with mitral valve reconstruction had lower short-term complication or death rates than did patients with prosthetic valve replacement (odds ratio 0.43, confidence interval 0.20-0.90). Eighty-two percent of patients with mitral valve reconstruction had no insufficiency or only trace insufficiency during the long-term follow-up period. Five-year complication free survivals were 64% (confidence interval 54%-74%) for patients undergoing mitral valve reconstruction and 47% (confidence interval 33%-60%) for patients undergoing prosthetic valve replacement. Results of a series of statistical analyses suggest that outcome was linked primarily to preoperative New York Heart Association functional class. CONCLUSIONS: Initial mortalities were similar among patients undergoing prosthetic replacement and valve reconstruction. Poor outcome was primarily related to preexisting comorbidities. Patients undergoing valve reconstruction had fewer valve-related complications. Valve reconstruction resulted in excellent durability and freedom from complications. These findings suggest that mitral valve reconstruction should be considered for appropriate patients with ischemic mitral insufficiency. PMID- 11726887 TI - Is repair preferable to replacement for ischemic mitral regurgitation? AB - OBJECTIVE: This study was undertaken to compare mitral valve repair and replacement as treatments for ischemic mitral regurgitation. METHODS: From 1985 through 1997, a total of 482 patients with ischemic mitral regurgitation underwent either valve repair (n = 397) or valve replacement (n = 85). Patients more likely (P < or =.01) to undergo repair had functional mitral regurgitation or coronary revascularization with an internal thoracic artery graft; those more likely to receive valve replacement were in higher New York Heart Association functional classes or underwent emergency operations. These factors were used for multivariable propensity matching. Risk factors for early and late death were identified by multivariable, multiphase hazard function analysis. RESULTS: Within the propensity-matched better-risk group, survivals after valve replacement were 81%, 56%, and 36% at 30 days, 1 year, and 5 years, but survivals after repair were 94%, 82%, and 58% at these intervals (P =.08). In contrast, within the poor risk group, survivals after repair and replacement were similar (P =.4). Risk factors (P < or =.01) included older age, higher functional class, greater wall motion abnormality, and renal dysfunction. Approximately 70% of patients were predicted to benefit from repair; the benefit lessened or was negated if an internal thoracic artery graft was not used, if a lateral wall motion abnormality was present, or if the mitral regurgitation jet pattern was complex. Freedom from repair failure at 5 years was 91%. CONCLUSION: Late survival is poor after surgery for ischemic mitral regurgitation. Most patients with ischemic mitral regurgitation benefit from mitral valve repair. In the most complex, high-risk settings, survivals after repair and replacement are similar. PMID- 11726889 TI - Intraoperative echocardiographic imaging of coronary arteries and graft anastomoses during coronary artery bypass grafting without cardiopulmonary bypass. AB - BACKGROUND: No accepted approach exists for the intraoperative evaluation of the quality of coronary arteries and the technical adequacy of graft anastomoses during coronary artery bypass grafting without cardiopulmonary bypass. METHODS AND RESULTS: We assessed the accuracy of high-frequency epicardial echocardiography and power Doppler imaging in evaluating coronary arteries during coronary artery bypass grafting without cardiopulmonary bypass. To validate measurements of coronary arteries and graft anastomoses by high-frequency epicardial echocardiography and power Doppler imaging, we compared luminal diameters determined by these methods with diameters determined histologically in a study of off-pump coronary artery bypass grafting in 20 dogs. Technical errors were deliberately created in 10 grafts (stenosis group). The results of these animal validation studies showed that the maximum luminal diameters of coronary arteries and graft anastomoses measured by high-frequency epicardial echocardiography (HEE) and power Doppler imaging (PDI) correlated well with the histologic measurements: HEE = 1.027 x Histologic measurements + 0.005 (P <.0001); PDI = 0.886 x Histologic measurements + 0.0453 (P =.0001). Similar results were found in the evaluation of the stenosis group: PDI = 0.991 x Histologic measurements + 0.074 (P <.0001). Subsequently, we demonstrated the clinical applicability of this approach in 12 patients who underwent minimally invasive or off-pump coronary artery bypass grafting. Twenty graft anastomoses were examined intraoperatively by high-frequency epicardial echocardiography and power Doppler imaging, and luminal diameters determined by power Doppler imaging were compared with those determined by postoperative coronary angiography. The results demonstrated that graft anastomosis by power Doppler imaging correlated well with the angiographic measurements: PDI = 1.018 x Angiographic measurements 0.106 (P <.0001). CONCLUSION: High-frequency epicardial echocardiography can provide meaningful information on the target coronary artery, and power Doppler imaging can accurately measure graft anastomoses and can detect technical errors and inadequacies during coronary artery bypass grafting without cardiopulmonary bypass. PMID- 11726888 TI - Microembolic signal counts increase during hyperbaric exposure in patients with prosthetic heart valves. AB - BACKGROUND: Patients with prosthetic heart valves have an increased risk of thromboembolic events, and transcranial Doppler sonography reveals microembolic signals. Whereas microembolic signals were initially assumed to be of particulate matter, recent studies suggest that they are partially gaseous in origin. If this is true, alteration of environmental pressure should change microembolic signal counts. We undertook this study to evaluate the influence of hyperbaric exposure on microembolic signal counts in persons with prosthetic heart valves. METHODS AND RESULTS: Microembolic signal counts were monitored by transcranial Doppler sonography of both middle cerebral arteries under normobaria (normobaria 1), 2 subsequent periods of hyperbaria (2.5 and 1.75 bar), and a second period of normobaria (normobaria 2) in 15 patients with prosthetic heart valves. Each monitoring period lasted 30 minutes. Compression and decompression rates were 0.1 bar/min. Microembolic signal counts increased from 20 (12-78) at normobaria 1 to 79 (30-165) at 2.5 bar (P <.01 vs normobaria 1 and 2), decreased to 44 (18-128) at 1.75 bar (P <.01 vs normobaria 1 and 2.5 bar; P <.001 vs normobaria 2), and returned to 20 (8-96) at normobaria 2 (values are medians and 95% confidence intervals). CONCLUSIONS: Our results strongly suggest that gaseous bubbles are underlying material for part of the microembolic signals detected in patients with prosthetic heart valves. PMID- 11726890 TI - Trimetazidine protects the energy status after ischemia and reduces reperfusion injury in a rat single lung transplant model. AB - BACKGROUND: Ischemia-reperfusion injury involves free radical generation and polymorphonuclear neutrophil chemotaxis. Trimetazidine is an anti-ischemic drug that restores the ability of the ischemic cells to produce energy and reduces the generation of oxygen-derived free radicals. We evaluated the effect of trimetazidine against ischemia-reperfusion injury after lung transplantation. METHODS: Rat single lung transplantation was performed in 3 experimental groups (n = 5): (1) the immediate transplantation group was defined as animals undergoing transplantation immediately after harvest without treatment; (2) the ischemic control group was defined as animals undergoing transplantation after 18 hours of cold (4 degrees C) ischemia without treatment; and (3) the trimetazidine treated group was defined as animals undergoing transplantation after 18 hours of cold (4 degrees C) ischemia and donor and recipient treatment with 5 mg/kg intravenous trimetazidine 10 minutes before harvest and reperfusion, respectively. All donor lungs were flushed with low-potassium dextran-glucose solution. After 2 hours of reperfusion, oxygenation was measured, and lung tissue was frozen and assessed for adenosine triphosphate content, myeloperoxidase activity, and thiobarbituric acid-reactive substances. Peak airway pressure was recorded throughout the reperfusion period. RESULTS: The trimetazidine group showed significantly higher levels of adenosine triphosphate content (1.73 +/- 0.8 pmol vs 0.72 +/- 0.2 pmol [ischemic control], P =.008), better oxygenation (238.82 +/- 113.9 mm Hg vs 89.39 +/- 14.7 mm Hg [ischemic control], P =.008), and reduced lipid peroxidation (1.28 +/- 0.1 nmol/g vs 2.09 +/- 0.4 nmol/g [ischemic control], P =.008). Adenosine triphosphate levels of the trimetazidine group were comparable with those of the immediate transplantation group (1.73 +/- 0.8 pmol vs 1.89 +/- 0.5 pmol, respectively; P =.31). Peak airway pressure and myeloperoxidase activity were comparable among groups. CONCLUSION: Donor and recipient treatment with trimetazidine provided a significant protection of the energy status, better oxygenation, and reduced lipid peroxidation in this experimental model. Our data suggest that trimetazidine may be an important adjunct to prolong ischemic time safely and to decrease lung ischemia-reperfusion injury. PMID- 11726891 TI - Expression of chemokine receptors CXCR1 and CXCR2 during cardiopulmonary bypass. AB - OBJECTIVE: This study investigated the effects of cardiopulmonary bypass on neutrophil expression of chemokine receptors, CXCR1 and CXCR2, and the beta2 integrin CD11b. METHODS: Ten patients undergoing coronary artery grafting with cardiopulmonary bypass were studied. Blood samples were collected preoperatively, before bypass, at termination of bypass, and 12 to 18 hours postoperatively. In vitro studies were performed on control subjects to determine changes in the surface expression of CXCR1, CXCR2, and CD11b on stimulation with interleukin 8. Receptor expression was measured by flow cytometry. Plasma levels of interleukin 8 from the patients were determined by enzyme-linked immunoassay. RESULTS: After bypass, CXCR2 expression fell by 66% (P <.0001) and remained low postoperatively (P <.0001). CXCR1 expression persisted at preoperative levels. CD11b expression increased significantly after bypass (P <.0001), returning to prebypass levels postoperatively. In vitro studies showed a dose-related fall of both CXCR1 (P <.0001) and CXCR2 expression (P <.0001) and a significant rise in CD11b expression (P <.0001). Plasma interleukin 8 increased significantly after bypass (P <.0001), remaining elevated 12 to 18 hours postoperatively (P =.02). Correlations between interleukin 8 levels and CXCR2 expression (P <.0001) and CD11b expression (P <.03) were demonstrated. CONCLUSIONS: CXCR2 expression is significantly down-regulated after bypass; in contrast, CXCR1 expression remains unchanged. In addition, whereas interleukin 8 is an important determinant of both CXCR1 and CXCR2 expression in vitro, it only correlates with CXCR2 and CD11b expression in vivo. This has implications in the search for antagonists against CXC chemokines and their receptors. PMID- 11726892 TI - Role of endogenous endothelin on coronary reflow after cardioplegic arrest. AB - OBJECTIVE: Endothelin plays a role in the regulation of basal coronary tone. We hypothesized that low coronary reflow and reduced cardiac function after prolonged ischemia may be due to increased release of endogenous endothelin. METHODS: Using an isolated perfused rat heart, we examined the effect of the addition of various endothelin antagonists during reperfusion after 4 hours of cardioplegic arrest at 4 degrees C. Hearts were freeze-clamped at the end of reperfusion for analysis of high-energy phosphate levels. Results are expressed as the percentages of preischemic values. RESULTS: The addition of bosentan or Ro61-0612 (nonselective endothelin antagonists) resulted in a significant increase in the recovery of coronary flow after 30 minutes of reperfusion (100.9% vs 85.3% [P =.03] and 122.4% vs 83.7% [P <.001], respectively, versus controls). The addition of PD155080 (endothelin A antagonist) had a similar effect (129.5% vs 91.4%, P =.008). BQ788 (endothelin B antagonist) and phosphoramidon (endothelin-converting enzyme inhibitor) had no effect. Myocardial adenosine triphosphate levels were significantly (12.1%) higher after reperfusion with Ro61 0612 (18.1 +/- 0.4 micromol/g vs 16.2 +/- 0.5 micromol/g, P =.01). There was no difference in the recovery of cardiac mechanical function with any of the antagonists studied. CONCLUSION: These results suggest that endogenous endothelin plays a role in low coronary reflow after prolonged cardioplegic arrest but does not impair recovery of myocardial function. PMID- 11726893 TI - Coronary endothelial dysfunction as a predictor of intimal thickening in the long term after heart transplantation. AB - OBJECTIVES: The mechanisms of cardiac allograft vasculopathy and its predisposing factors are multifactorial and as yet not well established. To determine the influence of endothelial dysfunction on the development of intimal thickening, we prospectively analyzed the vasomotor response to acetylcholine and nitroglycerin, as well as other donor and recipient variables. Findings were correlated with the coronary intimal thickness, which was evaluated by means of intravascular ultrasonography. METHODS: Nineteen patients who had undergone heart transplantation 4.89 +/- 2.35 years previously and who had angiographically normal coronary arteries were included. Endothelial function was analyzed by quantitative coronary analysis of the vasomotor response of the left anterior descending artery to acetylcholine. An intimal thickness index, reflecting the percentage of intima obstructing the coronary lumen, was calculated. RESULTS: Nine (47%) patients showed endothelial dysfunction, and the remaining 10 (53%) patients had a normal response. Four (44%) of 9 patients with a weight gain of greater than 20% after the operation showed endothelial dysfunction compared with none of the 10 patients with normal responses (P <.04). The severity of the intimal thickness correlated with the years after transplant (r = 0.45, P <.05). Patients with endothelial dysfunction had more intimal thickening than those without (32% +/- 17% vs 17% +/- 12%, respectively; P <.05). Furthermore, the degree of intimal thickening correlated with the magnitude of the vasomotor response to acetylcholine (r = -0.60, P =.006). No relationship was found between intimal thickness and the vasodilatory response to nitroglycerin. As independent variables for intimal thickness, multivariate analysis detected the magnitude of the response to acetylcholine (P =.0005), years after transplant (P =.01), and ischemic time (P =.03). CONCLUSIONS: Cardiac allograft vasculopathy is a multifactorial disease the severity of which increases over time. Endothelial dysfunction is a predictive factor of intimal thickening severity. Predisposing factors that provoke endothelial injury, such as perioperative ischemic time and obesity, may contribute to the development of allograft vasculopathy. PMID- 11726894 TI - Perioperative use of tirofiban hydrochloride (Aggrastat) does not increase surgical bleeding after emergency or urgent coronary artery bypass grafting. AB - BACKGROUND: The platelet glycoprotein IIb/IIIa inhibitor tirofiban hydrochloride improves outcome in patients with acute coronary syndrome. Nevertheless, a considerable number of patients require emergency or urgent coronary artery bypass grafting and may be at increased risk of postoperative bleeding after treatment with this molecule. The aim of this study is to evaluate the incidence of bleeding complications among patients undergoing bypass grafting after treatment with tirofiban. METHODS: We investigated the influence of the molecule on postoperative bleeding after cardiac surgery, comparing 2 groups of patients undergoing emergency or urgent coronary artery bypass grafting: group A (n = 20) received tirofiban, and group B (n = 68) received conventional therapy with intravenous heparin up until the operation. A total of 88 patients underwent coronary artery bypass surgery within 2 hours of ceasing the hemodynamic study. Clinical outcome, chest tube outputs, bleeding complications, transfusion requirements, platelet and hemoglobin counts, and clinical complications were examined. RESULTS: Bleeding differences were noted between the 2 groups at 8, 16, and 24 hours postoperatively. The incidence of blood, platelet, and fresh frozen plasma transfusions was higher in the control group. Postoperative thrombocytopenia was preserved in group A (199.5 +/- 70.4 vs 150.6 +/- 33.4 10(3)/mL, P <.01). No significant differences were noted between the 2 groups in the incidence of perioperative myocardial infarction, but significant differences were noted in enzyme levels, length of stay in the intensive care unit, and length of stay in the hospital. No deaths were observed. Hospital morbidity was increased in group B because of factors that were not apparently linked with tirofiban infusion. CONCLUSIONS: Patients may safely undergo coronary artery bypass surgery after treatment with tirofiban hydrochloride. This molecule, administered in the immediate preoperative period, has no adverse clinical effects and does not seem to negatively influence the incidence of perioperative myocardial infarction. Although extracorporeal circulation can modify platelet numbers and function, our ongoing data could show significant reduction in the loss of platelets induced by cardiopulmonary bypass, minor postoperative bleeding, and a minor transfusion requirement in general. PMID- 11726896 TI - Video-assisted minimally invasive approach in clinical bilateral lung transplantation. PMID- 11726895 TI - Multicenter clinical evaluation of the HeartMate vented electric left ventricular assist system in patients awaiting heart transplantation. AB - BACKGROUND: Despite advances in heart transplantation and mechanical circulatory support, mortality among transplant candidates remains high. Better ways are needed to ensure the survival of transplant candidates both inside and outside the hospital. METHODS: In a prospective, multicenter clinical trial conducted at 24 centers in the United States, 280 transplant candidates (232 men, 48 women; median age, 55 years; range, 11-72 years) unresponsive to inotropic drugs, intra aortic balloon counterpulsation, or both, were treated with the HeartMate Vented Electric Left Ventricular Assist System (VE LVAS). A cohort of 48 patients (40 men, 8 women; median age, 50 years; range, 21-67 years) not supported with an LVAS served as a historical control group. Outcomes were measured in terms of laboratory data (hemodynamic, hematologic, and biochemical), adverse events, New York Heart Association functional class, and survival. RESULTS: The VE LVAS treated and non-VE LVAS-treated (control) groups were similar in terms of age, sex, and distribution of patients by diagnosis (ischemic cardiomyopathy, idiopathic cardiomyopathy, and subacute myocardial infarction). VE LVAS support lasted an average of 112 days (range, < 1-691 days), with 54 patients supported for > 180 days. Mean VE LVAS flow (expressed as pump index) throughout support was 2.8 L x min(-1) x m(-2). Median total bilirubin values decreased from 1.2 mg/dL at baseline to 0.7 mg/dL (P =.0001); median creatinine values decreased from 1.5 mg/dL at baseline to 1.1 mg/dL (P =.0001). VE LVAS-related adverse events included bleeding in 31 patients (11%), infection in 113 (40%), neurologic dysfunction in 14 (5%), and thromboembolic events in 17 (6%). A total of 160 (58%) patients were enrolled in a hospital release program. Twenty-nine percent of the VE LVAS-treated patients (82/280) died before receiving a transplant, compared with 67% of controls (32/48) (P <.001). Conversely, 71% of the VE LVAS treated patients (198/280) survived: 67% (188/280) ultimately received a heart transplant, and 4% (10/280) had the device removed electively. One-year post transplant survival of VE LVAS-treated patients was significantly better than that of controls (84% [158/188] vs 63% [10/16]; log rank analysis P =.0197). CONCLUSION: The HeartMate VE LVAS provides adequate hemodynamic support, has an acceptably low incidence of adverse effects, and improves survival in heart transplant candidates both inside and outside the hospital. The studies of the HeartMate LVAS (both pneumatic and electric) for Food and Drug Administration approval are the only studies with a valid control group to show a survival benefit for cardiac transplantation. PMID- 11726897 TI - Biventricular repair of transposition of the great arteries and unbalanced ventricles. AB - BACKGROUND: It is well established that the arterial switch operation is the surgical procedure of choice in patients with transposition of the great arteries and balanced ventricular anatomy. The surgical approach of choice in patients with transposition but unbalanced ventricular size is unknown. OBJECTIVES: Since the beginning of the arterial switch operation program, patients with transposition of the great arteries and unbalanced ventricles underwent biventricular repair by means of the arterial switch operation and repair of any associated lesions, either through a single or staged surgical procedure. The aim of this retrospective study is to analyze whether this approach can be proposed to such patients. METHODS: Forty-four patients with transposition of the great arteries and unbalanced ventricles underwent this surgical approach since 1984. Two groups were defined: group I had transposition with a dominant right ventricle (n = 28), and group II had transposition with a dominant left ventricle (n = 16). In group I the median age and weight at the arterial switch operation were 8.5 days (range, 5-70 days) and 3.1 kg (range, 1.5-3.7 kg), respectively. The median end-diastolic left ventricular volume, mass, and long-axis ratio were 15 mL/m2 (range, 11-16 mL/m2), 31.5 g/m2 (range, 20-66 g/m2), and 0.85 (range, 0.9-0.7), respectively. The mitral valve diameter was slightly hypoplastic, with a median z value of -1.22 (range, -0.3 to 3.7). In group 2 the median age and weight at the arterial switch operation were 42 days (range, 8 days-15 years) and 3.5 kg (range, 2.8-35 kg), respectively. Associated lesions in this group were coarctation in 9 and single (n = 12) or multiple (n = 4) ventricular septal defects. The median long-axis ratio and tricuspid z value were 0.6 (range, 0.3 0.8) and -0.9 (range, -0.5 to 3.3), respectively. In this group 9 patients had a single-stage procedure with fenestrated ventricular defect patches, atrial septal defect patches, or both; 7 patients underwent the staged approach. RESULTS: In group I there was 1 early death from sepsis after weaning from postoperative extracorporeal membrane oxygenation. Three patients had severe pulmonary hypertension, one of whom died 1 year later. All survivors demonstrated, at discharge from the hospital, equilibrated ventricular size, with a median left ventricular end-diastolic volume of 25 mL/m2 (range, 21-30 mL/m2). In group II there were 2 early and 1 late deaths. All early deaths occurred in patients without voluntary residual intracardiac shunts. Median early postoperative long axis ratio and tricuspid z value were 0.8 (range, 0.7-1) and -0.2 (range, 0.74 to 1.2), respectively. CONCLUSION: This study demonstrates that the arterial switch operation in patients with transposition of the great arteries and unbalanced ventricles remains a good surgical option. PMID- 11726898 TI - Reconstruction of the right ventricular outflow tract with a bovine jugular vein graft fixed with a naturally occurring crosslinking agent (genipin) in a canine model. AB - OBJECTIVE: This study was designed to evaluate a newly developed biologic valved conduit fixed with genipin used to reconstruct the right ventricular outflow tract in a canine model. METHODS: Fresh bovine jugular veins with a retained native valve procured from a slaughterhouse were used as raw materials to fabricate the valved conduits. A naturally occurring crosslinking agent, genipin, was used to fix the procured jugular veins. The glutaraldehyde-fixed counterpart was used as a control. A canine model was used in the study. RESULTS: Echocardiography revealed that the motion of the valvular leaflets in both the glutaraldehyde- and genipin-fixed conduits was satisfactory. The transvalvular pressure gradients of both studied groups were minimal. No endothelium-like cells were observed on the luminal surface of the conduit and the valvular leaflet for the glutaraldehyde-fixed group throughout the entire course of the study. In contrast, endothelium-like cells were observed on the entire surface of the genipin-fixed valved conduit retrieved at 6 months postoperatively in all the cases studied. There was no evidence of luminal fibrous peel in any the valved conduits studied. Degradation of valvular leaflet in one of the glutaraldehyde fixed conduits was observed. In this particular case, thrombus formation was also observed on the surface of the valvular leaflet. On the other hand, no apparent degradation or thrombus formation was observed on the surfaces of the genipin fixed valvular leaflet and conduit. A significantly more severe inflammatory reaction was observed for the glutaraldehyde-fixed conduit than for its genipin fixed counterpart throughout the entire course of the study. The calcium contents of the samples before implantation and those retrieved at distinct implantation duration were minimal for both the glutaraldehyde- and genipin-fixed tissues. CONCLUSION: Although further studies are necessary, the genipin-fixed valved conduit appears to have great potential in helping mitigate the complications observed in the commercially available conduits. PMID- 11726899 TI - Extracardiac conduit versus lateral tunnel cavopulmonary connections at a single institution: impact on outcomes. AB - OBJECTIVE: To compare outcomes of extracardiac conduit and lateral tunnel Fontan connections in a single institution over a concurrent time period. METHODS: Between January 1994 and September 1998, 60 extracardiac conduit and 47 lateral tunnel total cavopulmonary connections were performed. Age, sex, and weight did not differ between the 2 groups. Compared with the lateral tunnel group (LT group), patients undergoing the extracardiac conduit procedure (EC group) had a trend to a higher incidence of morphologically right ventricle (EC group 48% vs LT group 32%; P <.09), a higher incidence of isomerism/heterotaxy syndrome (EC 22% vs LT 0%; P <.001), worse atrioventricular valve regurgitation (EC 11% moderate-plus vs LT 0%; P <.06), and lower McGoon indices (EC 1.8 +/- 0.5 vs LT 2.1 +/- 0.5; P <.03). Preoperative arrhythmias, transpulmonary gradients, room air oxygen saturations, ejection fractions, ventricular end-diastolic pressure, and pulmonary artery distortion did not differ between groups. Cardiopulmonary bypass times and fenestration usage were similar in both groups. RESULTS: Overall operative mortality was 5.6% and did not differ between groups. The LT group had a significantly higher incidence of postoperative sinoatrial node dysfunction (45% vs EC group 15%; P <.007), supraventricular tachycardia (33% vs EC group 8%; P <.0009), and need for temporary postoperative pacing (32% vs 12%; P <.01). Median duration of intensive care unit stay (EC 2 days, range 1-10 days, vs LT 2.8 days, range 1-103 days; P <.07) and ventilatory support (EC 1 day, range 0.25 10 days, vs LT 1 day, range 0.25-99 days; P <.03) were all longer in the LT group. Median chest tube drainage (EC 8 days, LT 9 days) was similar in both groups. Follow-up averaged 2.5 +/- 1.4 years in the EC group and 2.8 +/- 1.9 years in the LT group. There were 2 late deaths. Overall survival is 94% at 1 month, 92% at 1 year, and 92% at 5 years. Late ejection fraction or atrioventricular valve function did not differ between groups. Intermediate follow-up Holter analysis showed a higher incidence of atrial arrhythmias in the LT group (23% vs 7%; P <.02). Multivariable analysis showed that (1) prolonged cardiopulmonary bypass time was the only independent predictor for perioperative mortality, prolonged ventilation and intensive care unit length of stay, and increased time to final removal of chest tube drains and (2) lateral tunnel Fontan connection is an independent predictor of early postoperative and intermediate atrial arrhythmias. CONCLUSIONS: Although patients in the EC group were at higher preoperative risk, their outcomes were comparable with those of the LT group. Use of the extracardiac conduit technique for the modified Fontan operation reduces the risk of early and midterm atrial arrhythmia. PMID- 11726900 TI - Usefulness of continuous monitoring of airway resistance and flow-volume curve in the perioperative management of infants with central airway obstruction: a case of vascular ring. PMID- 11726901 TI - An ectopic thymoma with spontaneous infarction. PMID- 11726902 TI - Successful immediate phrenic nerve reconstruction during mediastinal tumor resection. PMID- 11726903 TI - Anatomic correction of Ebstein anomaly. PMID- 11726904 TI - Esophageal injury during radiofrequency ablation for atrial fibrillation. PMID- 11726905 TI - Paraplegia after open surgery using endovascular stent graft for aortic arch aneurysm. PMID- 11726906 TI - Intraventricular mitral annuloplasty technique for use with repair of posterior left ventricular aneurysm. PMID- 11726907 TI - Coronary artery bypass grafting in a 26-year-old man with total occlusion of the left main coronary artery related to Behcet disease. PMID- 11726908 TI - Reoperation for dilatation of the pulmonary autograft after the Ross procedure. PMID- 11726909 TI - Chylothorax: an unusual manifestation of a large atrial septal defect. PMID- 11726910 TI - One-year experience with the platelet glycoprotein IIb/IIIa antagonist tirofiban and heparin during cardiopulmonary bypass in patients with heparin-induced thrombocytopenia type II. PMID- 11726911 TI - The use of argatroban as an anticoagulant for cardiopulmonary bypass in cardiac operations. PMID- 11726912 TI - Donor lung salvage after neurogenic pulmonary edema with the use of post transplant extracorporeal membrane oxygenation. PMID- 11726913 TI - Challenges for the 21st century. PMID- 11726914 TI - Sex, flies and microarrays. PMID- 11726915 TI - Light responses of a plastic plant. PMID- 11726917 TI - A brief on bacterial biofilms. PMID- 11726916 TI - Lost in translation. PMID- 11726921 TI - Show me the data! PMID- 11726920 TI - Minimum information about a microarray experiment (MIAME)-toward standards for microarray data. AB - Microarray analysis has become a widely used tool for the generation of gene expression data on a genomic scale. Although many significant results have been derived from microarray studies, one limitation has been the lack of standards for presenting and exchanging such data. Here we present a proposal, the Minimum Information About a Microarray Experiment (MIAME), that describes the minimum information required to ensure that microarray data can be easily interpreted and that results derived from its analysis can be independently verified. The ultimate goal of this work is to establish a standard for recording and reporting microarray-based gene expression data, which will in turn facilitate the establishment of databases and public repositories and enable the development of data analysis tools. With respect to MIAME, we concentrate on defining the content and structure of the necessary information rather than the technical format for capturing it. PMID- 11726922 TI - Conjugation between bacterial and mammalian cells. AB - Bacterial conjugation, in which DNA is transferred from one bacterium to another, was first reported in 1946 and found to be mediated by the F factor. Although the F and RK2/RP4 prototypic plasmids can mediate the transfer of DNA from bacteria to yeast, there has been no evidence of classical bacterial conjugation to higher eukaryotes. Here, I present evidence of such transfer, using Escherichia coli, the RK2 plasmid system and Chinese hamster ovary CHO K1 cells. PMID- 11726923 TI - Mutation of the ST7 tumor suppressor gene on 7q31.1 is rare in breast, ovarian and colorectal cancers. AB - The gene ST7 has recently been implicated as the broad-range tumor suppressor on human chromosome 7q31.1. We did not detect somatic mutations in ST7 in any of 149 primary ovarian, breast or colon carcinomas. These data suggest that epigenetic downregulation or haploinsufficiency, rather than somatic genetic alterations, may be the primary mechanism of abrogation of ST7 function in these tumor types. PMID- 11726924 TI - Absence of ST7 mutations in tumor-derived cell lines and tumors. AB - The gene ST7 has been proposed as the multi-tissue tumor-suppressor gene (TSG) at chromosome 7q31.1. However, we have sought and failed to detect the truncating mutations reported to exist in this gene. PMID- 11726925 TI - The contributions of sex, genotype and age to transcriptional variance in Drosophila melanogaster. AB - Here we present a statistically rigorous approach to quantifying microarray expression data that allows the relative effects of multiple classes of treatment to be compared and incorporates analytical methods that are common to quantitative genetics. From the magnitude of gene effects and contributions of variance components, we find that gene expression in adult flies is affected most strongly by sex, less so by genotype and only weakly by age (for 1- and 6-wk flies); in addition, sex x genotype interactions may be present for as much as 10% of the Drosophila transcriptome. This interpretation is compromised to some extent by statistical issues relating to power and experimental design. Nevertheless, we show that changes in expression as small as 1.2-fold can be highly significant. Genotypic contributions to transcriptional variance may be of a similar magnitude to those relating to some quantitative phenotypes and should be considered when assessing the significance of experimental treatments. PMID- 11726926 TI - Deletion of Pten in mouse brain causes seizures, ataxia and defects in soma size resembling Lhermitte-Duclos disease. AB - Initially identified in high-grade gliomas, mutations in the PTEN tumor suppressor are also found in many sporadic cancers and a few related autosomal dominant hamartoma syndromes. PTEN is a 3'-specific phosphatidylinositol-3,4,5 trisphosphate (PI(3,4,5)P3) phosphatase and functions as a negative regulator of PI3K signaling. We generated a tissue-specific deletion of the mouse homolog Pten to address its role in brain function. Mice homozygous for this deletion (PtenloxP/loxP;Gfap-cre), developed seizures and ataxia by 9 wk and died by 29 wk. Histological analysis showed brain enlargement in PtenloxP/loxP;Gfap-cre mice as a consequence of primary granule-cell dysplasia in the cerebellum and dentate gyrus. Pten mutant cells showed a cell-autonomous increase in soma size and elevated phosphorylation of Akt. These data represent the first evidence for the role of Pten and Akt in cell size regulation in mammals and provide an animal model for a human phakomatosis condition, Lhermitte-Duclos disease (LDD). PMID- 11726927 TI - Pten regulates neuronal soma size: a mouse model of Lhermitte-Duclos disease. AB - Somatic inactivation of PTEN occurs in different human tumors including glioblastoma, endometrial carcinoma and prostate carcinoma. Germline mutations in PTEN result in a range of phenotypic abnormalities that occur with variable penetrance, including neurological features such as macrocephaly, seizures, ataxia and Lhermitte-Duclos disease (also described as dysplastic gangliocytoma of the cerebellum). Homozygous deletion of Pten causes embryonic lethality in mice. To investigate function in the brain, we used Cre-loxP technology to selectively inactivate Pten in specific mouse neuronal populations. Loss of Pten resulted in progressive macrocephaly and seizures. Neurons lacking Pten expressed high levels of phosphorylated Akt and showed a progressive increase in soma size without evidence of abnormal proliferation. Cerebellar abnormalities closely resembled the histopathology of human Lhermitte-Duclos disease. These results indicate that Pten regulates neuronal size in vivo in a cell-autonomous manner and provide new insights into the etiology of Lhermitte-Duclos disease. PMID- 11726928 TI - Computational identification of promoters and first exons in the human genome. AB - The identification of promoters and first exons has been one of the most difficult problems in gene-finding. We present a set of discriminant functions that can recognize structural and compositional features such as CpG islands, promoter regions and first splice-donor sites. We explain the implementation of the discriminant functions into a decision tree that constitutes a new program called FirstEF. By using different models to predict CpG-related and non-CpG related first exons, we showed by cross-validation that the program could predict 86% of the first exons with 17% false positives. We also demonstrated the prediction accuracy of FirstEF at the genome level by applying it to the finished sequences of human chromosomes 21 and 22 as well as by comparing the predictions with the locations of the experimentally verified first exons. Finally, we present the analysis of the predicted first exons for all of the 24 chromosomes of the human genome. PMID- 11726929 TI - Genes required for ionizing radiation resistance in yeast. AB - The ability of Saccharomyces cerevisiae to tolerate ionizing radiation damage requires many DNA-repair and checkpoint genes, most having human orthologs. A genome-wide screen of diploid mutants homozygous with respect to deletions of 3,670 nonessential genes revealed 107 new loci that influence gamma-ray sensitivity. Many affect replication, recombination and checkpoint functions. Nearly 90% were sensitive to other agents, and most new genes could be assigned to the following functional groups: chromatin remodeling, chromosome segregation, nuclear pore formation, transcription, Golgi/vacuolar activities, ubiquitin mediated protein degradation, cytokinesis, mitochondrial activity and cell wall maintenance. Over 50% share homology with human genes, including 17 implicated in cancer, indicating that a large set of newly identified human genes may have related roles in the toleration of radiation damage. PMID- 11726930 TI - A QTL for flowering time in Arabidopsis reveals a novel allele of CRY2. AB - Variation of flowering time is found in the natural populations of many plant species. The underlying genetic variation, mostly of a quantitative nature, is presumed to reflect adaptations to different environments contributing to reproductive success. Analysis of natural variation for flowering time in Arabidopsis thaliana has identified several quantitative trait loci (QTL), which have yet to be characterized at the molecular level. A major environmental factor that determines flowering time is photoperiod or day length, the length of the light period, which changes across the year differently with geographical latitude. We identified the EDI locus as a QTL partly accounting for the difference in flowering response to the photoperiod between two Arabidopsis accessions: the laboratory strain Landsberg erecta (Ler), originating in Northern Europe, and Cvi, collected in the tropical Cape Verde Islands. Positional cloning of the EDI QTL showed it to be a novel allele of CRY2, encoding the blue-light photoreceptor cryptochrome-2 that has previously been shown to promote flowering in long-day (LD) photoperiods. We show that the unique EDI flowering phenotype results from a single amino-acid substitution that reduces the light-induced downregulation of CRY2 in plants grown under short photoperiods, leading to early flowering. PMID- 11726931 TI - Natural variation in light sensitivity of Arabidopsis. AB - Because plants depend on light for growth, their development and physiology must suit the particular light environment. Plants native to different environments show heritable, apparently adaptive, changes in their response to light. As a first step in unraveling the genetic and molecular basis of these naturally occurring differences, we have characterized intraspecific variation in a light dependent developmental process-seedling emergence. We examined 141 Arabidopsis thaliana accessions for their response to four light conditions, two hormone conditions and darkness. There was significant variation in all conditions, confirming that Arabidopsis is a rich source of natural genetic diversity. Hierarchical clustering revealed that some accessions had response patterns similar to known photoreceptor mutants, suggesting changes in specific signaling pathways. We found that the unusual far-red response of the Lm-2 accession is due to a single amino-acid change in the phytochrome A (PHYA) protein. This change stabilizes the light-labile PHYA protein in light and causes a 100-fold shift in the threshold for far-red light sensitivity. Purified recombinant Lm-2 PHYA also shows subtle photochemical differences and has a reduced capacity for autophosphorylation. These biochemical changes contrast with previously characterized natural alleles in loci controlling plant development, which result in altered gene expression or loss of gene function. PMID- 11726932 TI - A 11.7-kb deletion triggers intersexuality and polledness in goats. AB - Mammalian sex determination is governed by the presence of the sex determining region Y gene (SRY) on the Y chromosome. Familial cases of SRY-negative XX sex reversal are rare in humans, often hampering the discovery of new sex-determining genes. The mouse model is also insufficient to correctly apprehend the sex determination cascade, as the human pathway is much more sensitive to gene dosage. Other species might therefore be considered in this respect. In goats, the polled intersex syndrome (PIS) mutation associates polledness and intersexuality. The sex reversal affects exclusively the XX individuals in a recessive manner, whereas the absence of horns is dominant in both sexes. The syndrome is caused by an autosomal gene located at chromosome band 1q43 (ref. 9), shown to be homologous to human chromosome band 3q23 (ref. 10). Through a positional cloning approach, we demonstrate that the mutation underlying PIS is the deletion of a critical 11.7-kb DNA element containing mainly repetitive sequences. This deletion affects the transcription of at least two genes: PISRT1, encoding a 1.5-kb mRNA devoid of open reading frame (ORF), and FOXL2, recently shown to be responsible for blepharophimosis ptosis epicanthus inversus syndrome (BPES) in humans. These two genes are located 20 and 200 kb telomeric from the deletion, respectively. PMID- 11726933 TI - Genetic mapping with SNP markers in Drosophila. AB - Map-based positional cloning of Drosophila melanogaster genes is hampered by both the time-consuming, error-prone nature of traditional methods for genetic mapping and the difficulties in aligning the genetic and cytological maps with the genome sequence. The identification of sequence polymorphisms in the Drosophila genome will make it possible to map mutations directly to the genome sequence with high accuracy and resolution. Here we report the identification of 7,223 single nucleotide polymorphisms (SNPs) and 1,392 insertions/deletions (InDels) in common laboratory strains of Drosophila. These sequence polymorphisms define a map of 787 autosomal marker loci with a resolution of 114 kb. We have established PCR product-length polymorphism (PLP) or restriction fragment-length polymorphism (RFLP) assays for 215 of these markers. We demonstrate the use of this map by delimiting two mutations to intervals of 169 kb and 307 kb, respectively. Using a local high-density SNP map, we also mapped a third mutation to a resolution of approximately 2 kb, sufficient to localize the mutation within a single gene. These methods should accelerate the rate of positional cloning in Drosophila. PMID- 11726937 TI - Petrifying research? PMID- 11726938 TI - Cellular function of the Fanconi anemia pathway. PMID- 11726940 TI - US plans xenophobic science laws. PMID- 11726941 TI - Canada tops cancer research impact list. PMID- 11726942 TI - Patents not key barrier to AIDS therapy. PMID- 11726943 TI - NIH posts online ES-cell registry. PMID- 11726944 TI - New TB vaccines to be tested. PMID- 11726945 TI - Fifteen-year follow up for gene therapy patients. PMID- 11726946 TI - UK cancer charities concerned about new EU regulations. PMID- 11726948 TI - US to revise vaccine compensation program. PMID- 11726949 TI - US dilutes smallpox vaccine supplies. PMID- 11726950 TI - New class of HIV drugs shows promise. PMID- 11726951 TI - Rapid anthrax test developed. PMID- 11726952 TI - US cancer research strategy revised. PMID- 11726953 TI - Confusion reigns over breast cancer. PMID- 11726956 TI - Bioterrorism: a clear and present danger. PMID- 11726959 TI - Seeing death in the living. PMID- 11726960 TI - HIV-1 and Ebola virus: the getaway driver nabbed. PMID- 11726961 TI - alpha-synuclein and cytosolic dopamine: stabilizing a bad situation. PMID- 11726962 TI - A new feature on the cholesterol-lowering landscape. PMID- 11726964 TI - Annihilating host defense. PMID- 11726965 TI - Death by influenza virus protein. PMID- 11726967 TI - Research news. PMID- 11726966 TI - A fresh facet for ouabain action. PMID- 11726968 TI - Possible new role for NF-kappaB in the resolution of inflammation. AB - Inflammation involves the sequential activation of signaling pathways leading to the production of both pro- and anti-inflammatory mediators. Although much attention has focused on pro-inflammatory pathways that initiate inflammation, relatively little is known about the mechanisms that switch off inflammation and resolve the inflammatory response. The transcription factor NF-kappaB is thought to have a central role in the induction of pro-inflammatory gene expression and has attracted interest as a new target for the treatment of inflammatory disease. We show here that NF-kappaB activation in leukocytes recruited during the onset of inflammation is associated with pro-inflammatory gene expression, whereas such activation during the resolution of inflammation is associated with the expression of anti-inflammatory genes and the induction of apoptosis. Inhibition of NF-kappaB during the resolution of inflammation protracts the inflammatory response and prevents apoptosis. This suggests that NF-kappaB has an anti inflammatory role in vivo involving the regulation of inflammatory resolution. PMID- 11726969 TI - Evasion of human innate and acquired immunity by a bacterial homolog of CD11b that inhibits opsonophagocytosis. AB - Microbial pathogens must evade the human immune system to survive, disseminate and cause disease. By proteome analysis of the bacterium Group A Streptococcus (GAS), we identified a secreted protein with homology to the alpha-subunit of Mac 1, a leukocyte beta2 integrin required for innate immunity to invading microbes. The GAS Mac-1-like protein (Mac) was secreted by most pathogenic strains, produced in log-phase and controlled by the covR-covS two-component gene regulatory system, which also regulates transcription of other GAS virulence factors. Patients with GAS infection had titers of antibody specific to Mac that correlated with the course of disease, demonstrating that Mac was produced in vivo. Mac bound to CD16 (FcgammaRIIIB) on the surface of human polymorphonuclear leukocytes and inhibited opsonophagocytosis and production of reactive oxygen species, which resulted in significantly decreased pathogen killing. Thus, by mimicking a host-cell receptor required for an innate immune response, the GAS Mac protein inhibits professional phagocyte function by a novel strategy that enhances pathogen survival, establishment of infection and dissemination. PMID- 11726970 TI - A novel influenza A virus mitochondrial protein that induces cell death. AB - While searching for alternative reading-frame peptides encoded by influenza A virus that are recognized by CD8+ T cells, we found an abundant immunogenic peptide encoded by the +1 reading frame of PB1. This peptide derives from a novel conserved 87-residue protein, PB1-F2, which has several unusual features compared with other influenza gene products in addition to its mode of translation. These include its absence from some animal (particularly swine) influenza virus isolates, variable expression in individual infected cells, rapid proteasome dependent degradation and mitochondrial localization. Exposure of cells to a synthetic version of PB1-F2 induces apoptosis, and influenza viruses with targeted mutations that interfere with PB1-F2 expression induce less extensive apoptosis in human monocytic cells than those with intact PB1-F2. We propose that PB1-F2 functions to kill host immune cells responding to influenza virus infection. PMID- 11726971 TI - HIV-1 and Ebola virus encode small peptide motifs that recruit Tsg101 to sites of particle assembly to facilitate egress. AB - Retroviral Gag proteins encode sequences, termed late domains, which facilitate the final stages of particle budding from the plasma membrane. We report here that interactions between Tsg101, a factor involved in endosomal protein sorting, and short peptide motifs in the HIV-1 Gag late domain and Ebola virus matrix (EbVp40) proteins are essential for efficient egress of HIV-1 virions and Ebola virus-like particles. EbVp40 recruits Tsg101 to sites of particle assembly and a short, EbVp40-derived Tsg101-binding peptide sequence can functionally substitute for the HIV-1 Gag late domain. Notably, recruitment of Tsg101 to assembling virions restores budding competence to a late-domain-defective HIV-1 in the complete absence of viral late domain. These studies define an essential virus host interaction that is conserved in two unrelated viruses. Because the Tsg101 is recruited by small, conserved viral sequence motifs, agents that mimic these structures are potential inhibitors of the replication of these lethal human pathogens. PMID- 11726972 TI - Mucosal AIDS vaccine reduces disease and viral load in gut reservoir and blood after mucosal infection of macaques. AB - Given the mucosal transmission of HIV-1, we compared whether a mucosal vaccine could induce mucosal cytotoxic T lymphocytes (CTLs) and protect rhesus macaques against mucosal infection with simian/human immunodeficiency virus (SHIV) more effectively than the same vaccine given subcutaneously. Here we show that mucosal CTLs specific for simian immunodeficiency virus can be induced by intrarectal immunization of macaques with a synthetic-peptide vaccine incorporating the LT(R192G) adjuvant. This response correlated with the level of T-helper response. After intrarectal challenge with pathogenic SHIV-Ku2, viral titers were eliminated more completely (to undetectable levels) both in blood and intestine, a major reservoir for virus replication, in intrarectally immunized animals than in subcutaneously immunized or control macaques. Moreover, CD4+ T cells were better preserved. Thus, induction of CTLs in the intestinal mucosa, a key site of virus replication, with a mucosal AIDS vaccine ameliorates infection by SHIV in non-human primates. PMID- 11726973 TI - HIV protease inhibitors protect apolipoprotein B from degradation by the proteasome: a potential mechanism for protease inhibitor-induced hyperlipidemia. AB - Highly active anti-retroviral therapies, which incorporate HIV protease inhibitors, resolve many AIDS-defining illnesses. However, patients receiving protease inhibitors develop a marked lipodystrophy and hyperlipidemia. Using cultured human and rat hepatoma cells and primary hepatocytes from transgenic mice, we demonstrate that protease inhibitor treatment inhibits proteasomal degradation of nascent apolipoprotein B, the principal protein component of triglyceride and cholesterol-rich plasma lipoproteins. Unexpectedly, protease inhibitors also inhibited the secretion of apolipoprotein B. This was associated with inhibition of cholesteryl-ester synthesis and microsomal triglyceride transfer-protein activity. However, in the presence of oleic acid, which stimulates neutral-lipid biosynthesis, protease-inhibitor treatment increased secretion of apolipoprotein B-lipoproteins above controls. These findings suggest a molecular basis for protease-inhibitor-associated hyperlipidemia, a serious adverse effect of an otherwise efficacious treatment for HIV infection. PMID- 11726974 TI - SCAP ligands are potent new lipid-lowering drugs. AB - Upregulation of low-density lipoprotein receptor (LDLr) is a key mechanism to control elevated plasma LDL-cholesterol levels. Here we identify a new class of compounds that directly binds to the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP). We show that a 14C-labeled, photo activatable analog specifically labeled both SCAP and a truncated form of SCAP containing the sterol-sensing domain. When administered to hyperlipidemic hamsters, SCAP ligands reduced both LDL cholesterol and triglycerides levels by up to 80% with a three-fold increase in LDLr mRNA in the livers. Using human hepatoma cells, we show that these compounds act through the sterol-responsive element of the LDLr promoter and activate the SCAP/SREBP pathway, leading to increased LDLr expression and activity, even in presence of excess of sterols. These findings have led to the identification of a class of compounds that represent a promising new class of hypolipidemic drugs. PMID- 11726975 TI - Stromal-derived factor-1 in human tumors recruits and alters the function of plasmacytoid precursor dendritic cells. AB - Dendritic-cell (DC) trafficking and function in tumors is poorly characterized, with studies confined to myeloid DCs (DC1s). Tumors inhibit DC1 migration and function, likely hindering specific immunity. The role of plasmacytoid DCs (DC2s) in tumor immunity is unknown. We show here that malignant human ovarian epithelial tumor cells express very high levels of stromal-derived factor-1, which induces DC2 precursor (preDC2) chemotaxis and adhesion/transmigration, upregulates preDC2 very late antigen (VLA)-5, and protects preDC2s from tumor macrophage interleukin-10-induced apoptosis, all through CXC chemokine receptor 4. The VLA-5 ligand vascular-cell adhesion molecule-1 mediated preDC2 adhesion/transmigration. Tumor preDC2s induced significant T-cell interleukin-10 unrelated to preDC2 differentiation or activation state, and this contributed to poor T-cell activation. Myeloid precursor DCs (preDC1s) were not detected. Tumors may weaken immunity by attracting preDC2s and protecting them from the harsh microenvironment, and by altering preDC1 distribution. PMID- 11726976 TI - Annexin-V imaging for noninvasive detection of cardiac allograft rejection. AB - Heart transplant rejection is characterized pathologically by myocyte necrosis and apoptosis associated with interstitial mononuclear cell infiltration. Any one of these components can be targeted for noninvasive detection of transplant rejection. During apoptotic cell death, phosphatidylserine, a phospholipid that is normally confined to the inner leaflet of cell membrane bilayer, gets exteriorized. Technetium-99m-labeled annexin-V, an endogenous protein that has high affinity for binding to phosphatidylserine, has been administered intravenously for noninvasive identification of apoptotic cell death. In the present study of 18 cardiac allograft recipients, 13 patients had negative and five had positive myocardial uptake of annexin. These latter five demonstrated at least moderate transplant rejection and caspase-3 staining, suggesting apoptosis in their biopsy specimens. This study reveals the clinical feasibility and safety of annexin-V imaging for noninvasive detection of transplant rejection by targeting cell membrane phospholipid alterations that are commonly associated with the process of apoptosis. PMID- 11726977 TI - Real-time imaging of apoptotic cell-membrane changes at the single-cell level in the beating murine heart. AB - We report a novel real-time imaging model to visualize apoptotic membrane changes of single cardiomyocytes in the injured heart of the living mouse, using fluorescent labeled annexin-V. Annexin-V binds to externalized phosphatidylserine (PS) of cells undergoing programmed cell death. With high-magnification (x100 160) real-time imaging, we visualized the binding of annexin-V to single cardiomyocytes. Kinetic studies at the single-cell level revealed that cardiomyocytes started to bind annexin-V within minutes after reperfusion, following an ischemic period of 30 minutes. The amount of bound annexin-V increased rapidly and reached a maximum within 20-25 minutes. Caspase inhibitors decreased the number of annexin-V-positive cardiomyocytes and slowed down the rate of PS exposure of cardiomyocytes that still bound annexin-V. This technology to study cell biology in the natural environment will enhance knowledge of intracellular signaling pathways relevant for cell-death regulation and strategies to manipulate these pathways for therapeutic effect. PMID- 11726978 TI - Targeting gene-modified hematopoietic cells to the central nervous system: use of green fluorescent protein uncovers microglial engraftment. AB - Gene therapy in the central nervous system (CNS) is hindered by the presence of the blood-brain barrier, which restricts access of serum constituents and peripheral cells to the brain parenchyma. Expression of exogenously administered genes in the CNS has been achieved in vivo using highly invasive routes, or ex vivo relying on the direct implantation of genetically modified cells into the brain. Here we provide evidence for a novel, noninvasive approach for targeting potential therapeutic factors to the CNS. Genetically-modified hematopoietic cells enter the CNS and differentiate into microglia after bone-marrow transplantation. Up to a quarter of the regional microglial population is donor derived by four months after transplantation. Microglial engraftment is enhanced by neuropathology, and gene-modified myeloid cells are specifically attracted to the sites of neuronal damage. Thus, microglia may serve as vehicles for gene delivery to the nervous system. PMID- 11726980 TI - Research update. PMID- 11726982 TI - This is so COOL. PMID- 11726983 TI - Canadian Cardiovascular Society Consensus Conference 2001 update: recommendations for the management of adults with congenital heart disease part III. PMID- 11726985 TI - Assessment of ventricular diastolic function. AB - A large number of patients suspected of having congestive heart failure have normal left ventricular systolic function and may, therefore, have primary diastolic heart failure. This diagnosis, however, should not be made unless there is also objective evidence of diastolic dysfunction, ie, signs of abnormal left ventricular relaxation and/or diastolic distensibility. The most useful noninvasive diagnostic approaches are the measurement of transmitral and pulmonary venous flow velocities by pulsed wave Doppler, and mitral annulus velocities by tissue Doppler echocardiography. In some patients, the assessment of intraventricular flow propagation by colour M-mode Doppler echocardiography provides additional information. Diastolic heart failure is most often due to coronary artery disease and/or hypertension; therefore, other noninvasive or invasive tests are needed to define the etiology of myocardial dysfunction. However, in the few patients who have constrictive pericarditis, the Doppler echocardiographic assessment of diastolic filling provides the most important clues to the etiology of the disease. Doppler echocardiographic assessment of left ventricular filling may also be used to obtain semiquantitative estimates of left ventricular diastolic pressure. Furthermore, left ventricular filling patterns, in particular, the deceleration time of early transmitral filling, are powerful predictors of patient prognosis. It is probably not cost effective to perform a comprehensive assessment of diastolic filling in every patient undergoing an echocardiographic examination. However, in selected patients, the assessment of diastolic filling provides information that is important for patient management. PMID- 11726984 TI - Altered metabolite concentrations with amiodarone generic substitution cannot be observed without monitoring. AB - The use of amiodarone has grown rapidly, resulting in the marketing of several generic formulations. The adequacy of the testing used to approve these formulations as bioequivalent has been questioned, and mounting clinical evidence suggests that in some patients, substitution with generic amiodarone can cause serious problems. The effects of switching amiodarone formulations may take weeks to develop, leaving the relationship between the events unrecognized. In animal models, the toxicity of desethylamiodarone, an active metabolite partly formed during amiodarone absorption, is greater than that of its parent compound. High metabolite to amiodarone ratios have been associated with clinical toxicity. Because measuring serum amiodarone and metabolite is not standard clinical practice, aberrations after switching formulations will be missed. Major changes in metabolite concentrations were documented in four patients switched to a generic formulation, suggesting that the tests used for regulatory approval failed to identify the cumulative effects of differing excipients on amiodarone metabolism during absorption. Physicians should monitor patients for several months after a switch in amiodarone formulation is made. Regulatory criteria for bioequivalence of amiodarone need to be reconsidered. PMID- 11726986 TI - The endocarditis that was not: an unusual case of heparin-induced thrombocytopenia with unusual complications. AB - The complications of heparin-induced thrombocytopenia have been well described previously. However, evidence of the possibility that heparin-induced thrombocytopenia can trigger a thyroid storm has never been published before. A catastrophic evolution of a man referred with a high endocarditis suspicion previously treated with heparin, who successively developed arterial thrombosis and thyroid storm, is described. PMID- 11726987 TI - Resection of renal cell carcinomas with inferior vena caval extension using deep hypothermic circulatory arrest. AB - Renal cell carcinoma with tumour thrombus extension into the inferior vena cava presents a difficult surgical challenge. The conventional surgical approach, which involves isolating the inferior vena cava, incising its wall and removing the thrombus, can have high incidences of perioperative mortality and embolization of the tumour thrombus compounded by severe hemorrhage. Four patients with renal cell carcinomas extending into the inferior vena cava were supported with cardiopulmonary bypass and deep hypothermic circulatory arrest during tumour excision. All of the operations were successfully performed with no mortality and minimal morbidity. The technique allowed the surgeon to operate in a bloodless field, thereby improving visibility and allowing complete tumour excision without significantly prolonging operative time. It is believed that this technique has improved the safety and technical feasibility of what had previously been a complicated and risky surgical procedure. PMID- 11726988 TI - Coexisting ventricular septal aneurysms: congenital and postmyocardial infarction. PMID- 11726990 TI - Canadian Cardiovascular Society 2000 Consensus Conference: Women and Ischemic Heart Disease. PMID- 11726989 TI - Reflections on the state of our health care system. PMID- 11726991 TI - Sex-related differences in the pathophysiology of cardiovascular disease: is there a rationale for sex-related treatments? PMID- 11726992 TI - Epidemiology of ischemic heart disease in women. PMID- 11726993 TI - Risk factors and primary prevention of ischemic heart disease in women. PMID- 11726994 TI - Hormone replacement therapy and cardiovascular disease. PMID- 11726995 TI - Clinical evaluation of women with ischemic heart disease: diagnosis and noninvasive testing. PMID- 11726996 TI - The medical management of acute coronary syndromes and chronic ischemic heart disease in women. PMID- 11726997 TI - Revascularization strategies in women with ischemic heart disease. PMID- 11726998 TI - Rehabilitation. PMID- 11726999 TI - Differences in access to care. PMID- 11727000 TI - Women and heart disease in Canada. PMID- 11727002 TI - Pyloric balloon dilation for delayed gastric emptying in children. AB - Delayed gastric emptying may manifest with symptoms of epigastric pain, early satiety and delayed vomiting, and at times may be associated with failure to thrive. These symptoms and signs may improve following surgical pyloroplasty. To determine whether pyloric balloon dilation (PBD) is an effective therapy for children with these symptoms, hospital records of all children who underwent endoscopic PBD between October 1991 and March 1994 at British Columbia's Children's Hospital were reviewed. Excluded were children with chromosomal abnormalities, neurological disorders and erosive esophagitis. Through-the-scope balloons of diameter 15 or 18 mm were positioned in the pyloric channel and inflated with air to 2334 or 1815 mmHg respectively, for 2 min. Nineteen children with a mean age of 3.75 years (range eight months to 10 years) who presented with symptoms for more than three months (mean 11 months) were identified. Eleven children presented with failure to thrive, 14 with delayed vomiting and 10 with early satiety. Results of gastric emptying tests at 90 min ranged from 8% to 75% (mean 32%). The pylorus was difficult to intubate in 11 of 19 children, and in two the pylorus could not be passed before PBD. No complications were experienced with PBD. Thirteen children had complete resolution of symptoms, and five had transient improvement lasting four to eight weeks after PBD with subsequent complete resolution of symptoms following surgical pyloroplasty. One child continued to have mild symptoms after PBD but did not have further treatment. This study suggests that PBD is a safe and effective therapeutic option in children with symptoms and signs associated with delayed gastric emptying. PMID- 11727003 TI - Liver transplantation: evolving patient selection criteria. AB - The widespread recognition of the success of liver transplantation as a treatment for most types of acute and chronic liver failure has led to increased referrals for transplantation in the setting of a relatively fixed supply of cadaver donor organs. These events have led to a marked lengthening of the waiting time for liver transplantation, resulting in increased deaths of those on the waiting list and sicker patients undergoing transplantation. Nearly 5000 liver transplantations were performed in the United States in 2000, while the waiting list grew to over 17,000 patients. The mounting disparity between the number of liver transplant candidates and the limited supply of donor organs has led to reassessment of the selection and listing criteria for liver transplantation, as well as revision of organ allocation and distribution policies for cadaver livers. The development of minimal listing criteria for patients with chronic liver disease based on a specific definition for decompensation of cirrhosis has facilitated the more uniform listing of patients at individual centres across the United States. The United Network for Organ Sharing, under pressure from transplant professionals, patient advocacy groups and the federal government, has continuously revised allocation and distribution policies based on the ethical principles of justice for the individual patient versus optimal utility of the limited organ supply available annually. Beginning in 2002, it is likely that the Model for End-stage Liver Disease (MELD) score will be implemented to determine disease severity and direct donor organs to the sickest patients rather than to those with the longest waiting times. PMID- 11727004 TI - An approach to iron-deficiency anemia. AB - Iron-deficiency anemia is a common reason for referral to a gastroenterologist. In adult men and postmenopausal women, gastrointestinal tract pathology is often the cause of iron-deficiency anemia, so patients are frequently referred for endoscopic evaluation. Endoscopy may be costly and at times difficult for the patient. Therefore, physicians need to know what lesions can be identified reliably and, more importantly, the importance of ruling out life-threatening conditions such as occult malignancy. Over the past decade, a number of prospective studies have been completed that examined the yield of endoscopy in the investigation of iron-deficiency anemia. The present article provides a broad overview of iron-deficiency anemia, with particular emphasis on hematological diagnosis, etiology, the use of endoscopy in identifying lesions and iron repletion therapy. Other clinical scenarios, including assessment of patients on anti-inflammatory or anticoagulation therapy and patients with bleeding of obscure origin, are also addressed. The present article provides a diagnostic algorithm to iron-deficiency anemia, which describes a more systematic manner in which to approach iron-deficiency anemia. PMID- 11727005 TI - Management of fistulas in patients with Crohn's disease: antibiotic to antibody. AB - Fistulas are common in patients with Crohn's disease and, when associated with inflammatory disease and established for several weeks, tend to be chronic. Perianal fistulas are the most frequent complication of, and are most often associated with, colonic disease. Perianal fistulas commonly require surgical resection and permanent ileostomy. Antibiotics, cyclosporine, methotrexate and thalidomide have been used in uncontrolled trials; only azathioprine, 6 mercaptopurine and infliximab have been assessed in double-blind, placebo controlled studies. Relapse of the fistula occurs with all drugs, unless treatment is continued long term. Each drug differs in its onset of action and long term tolerability. An approach to fistulizing disease in Crohn's disease is suggested. PMID- 11727006 TI - Familial occurrence of lymphocytic colitis. AB - The familial occurrence of lymphocytic colitis in a female parent and her two female children is reported. No other genetically based disorder, including celiac disease, was evident. For both children, the age of diagnosis was more than two decades younger than the age of recognition of disease in the parent, and some clinical features, including the requirement for pharmacological agents in both children, suggested that their disease severity was more significant than that of the involved parent. These characteristics of a familial disease have been previously reported and labelled 'genetic anticipation' in some monogenetic forms of neurological disease, as well as in other types of inflammatory bowel diseases, including Crohn's disease. Alternatively, a common cohort effect related to a pathological environmental factor may have played a role in the pathogenesis of this disorder. PMID- 11727007 TI - Osteonecrosis in a patient with Crohn's disease unrelated to corticosteroid use. AB - Osteonecrosis is a feared complication of corticosteroid use. However, a direct association between corticosteroid use and osteonecrosis has never been proven. The present report examines the case of a patient with longstanding Crohn's disease who had never been treated with corticosteroids and who developed osteonecrosis of the talus. The association of systemic inflammatory disorders with osteonecrosis and the possible association with vascular thrombosis are discussed. PMID- 11727009 TI - Embryo-maternal signaling prior to implantation. PMID- 11727010 TI - Investigation of an in vitro model of trophoblast invasion. AB - OBJECTIVES: To re-investigate methods for visualising cytotrophoblast invasion using the Matrigel invasion model. STUDY DESIGN: Cytotrophoblast cells were isolated from pooled first trimester placentae and cultured on Matrigel-coated transwells in media supplemented with either 5 ng/ml vascular endothelial growth factor (VEGF) or 10 ng/ml epidermal growth factor (EGF). Invasion was visualised using standard and confocal fluorescent microscopy. RESULTS: The purity of the enriched cytotrophoblast cells was 84-100%. Immunofluorescent cytokeratin staining examined with an inverted fluorescent microscope suggested that cytokeratin-positive cells were present on the underside of the membrane, having invaded through the Matrigel on the upper surface. However, confocal microscopy indicated that these cells did not invade through the Matrigel, and no viable cells were identified in the culture media below the membrane. CONCLUSION: These results suggest that cytokeratin-positive staining on Matrigel-coated transwells is not necessarily indicative of cell invasion, and that similar studies should be interpreted with caution depending on the method of quantification. PMID- 11727011 TI - Luteal estrogen supplementation in pregnancies associated with low serum estradiol concentrations. AB - The role of luteal phase estrogen in pregnancy outcome has been a matter of considerable debate. In order to evaluate the effectiveness of estrogen supplementation in gonadotropin releasing hormone agonist (GnRHa)/human menopausal gonadotropin (hMG)-stimulated cycles associated with low luteal estrogen concentration, a study was performed comparing the ongoing pregnancy rates in cycles with serum concentrations of estradiol (E2) <100 pg/ml 11 days post embryo transfer (p-ET), treated with luteal phase progesterone (P4) vs. E2 and P4 supplementation. Among 1106 serum samples studied, 951 were from women receiving GnRHa and follicle stimulating hormone (FSH) prior to oocyte retrieval and P4 (50 mg-100 mg IM daily) as luteal phase supplementation beginning day 11 after retrieval. The remaining 155 were from women receiving both E2 (2 mg-6 mg estrace orally each day) and P4 during the luteal phase. Significantly greater frequencies of preclinical losses were observed among women with human chorionic gonadotropin (hCG) concentrations>5 mIU/ml and concurrent E2 concentrations <100 pg/ml compared with E2 >100 pg/ml (p<0.00001). Among the 128 women who had hCG concentrations >5 mIU/ml and E2 concentrations <100 pg/ml, 102 received P4 only during the luteal phase and 26 were treated with estrace 2 mg-6 mg daily, as well as P4 during the luteal phase. The frequency of preclinical pregnancy losses among the 102 women with hCG >5 mIU/ml and E2 <100 pg/ml who did not receive luteal E2 supplementation was 72%, compared with 50% who received luteal E2 supplementation (p=0.04) The increase in preclinical pregnancy loss rates among women not receiving luteal E2 resulted in a decrease in ongoing pregnancy rate (8%), compared to those receiving luteal E2 supplementation (31%) (p=0.002). Our results indicated that a subset of women losing pregnancies preclinically after GnRHa and FSH stimulation due to low luteal phase serum E2 level may benefit from luteal estrogen supplementation. More sensitive and specific markers are needed to identify prospectively women in this risk group. PMID- 11727012 TI - Severe abdominal bleeding 51 days after laparoscopic salpingostomy for ectopic pregnancy: a case report. AB - Conservative management of ectopic pregnancy is important because it allows preservation of the fallopian tube. It has been reported to result in extratubal secondary trophoblastic implants (ESTI) in 3% to 22% of cases. The aim of this case report is to highlight the factors predicting the risk of ESTI. PMID- 11727013 TI - Ultrasonographic criteria for non-viability of first trimester intra-uterine pregnancy. AB - This prospective cohort study evaluated the performance of transvaginal sonography in the assessment of first trimester intra-uterine pregnancy. Consecutive women with singleton pregnancies who underwent a first trimester transvaginal sonographic examination between May 1995 and March 1996 were included. Women had to have an intra-uterine gestational sac and time since last menstrual period of less than 12 weeks. In case of absent cardiac activity and a crow-rump length < 12 mm, the test sonography was repeated with a 7 to 10 days interval. A final diagnosis, that was considered to be the gold standard in further analysis, was established by sonography performed one week after the last test sonography. Among 372 pregnancies, there were 92 (25%) non-viable. The combination of absence of cardiac activity and absence of a yolk sac appeared to be a highly specific criterion whenever the mean sac diameter was > or = 16 mm or whenever a crown rump length was > 5 mm. We conclude that single transvaginal sonography is a reliable test to diagnose non-viable pregnancy in case the MSD is > or = 16 mm or in case the crown rump length is > 5 mm. The combination of absence of cardiac activity and absence of a yolk sac virtually rules out the possibility of a viable pregnancy in these patients. PMID- 11727014 TI - Current progress in early pregnancy investigation. PMID- 11727015 TI - Detection of early atherosclerosis with radiolabeled monocyte chemoattractant protein-1 in prediabeteic Zucker rats. AB - BACKGROUND: Migration of monocytes into the arterial wall is an early finding of atherosclerosis. Monocytes are attracted to sites of vascular endothelial cell injury, the initiating event in the development of atheromatous disease, by a chemokine known as monocyte chemoattractant protein-1 (MCP-1). Injured vascular endothelial and smooth muscle cells selectively secrete MCP-1. OBJECTIVE: This study was performed to determine if radiolabeled MCP-1 would co-localize at sites of monocyte/macrophage concentration in an experimental model of transplant induced vasculopathy in diabetic animals. MATERIALS AND METHODS: Hearts from 3 month-old male Zucker rats, heterozygote (Lean) or homozygote (Fat) for the diabetes-associated gene fa, were transplanted into the abdomens of genetically matched recipients. Lean and Fat animals were then fed normal or high-fat diets for 90 days. RESULTS: At 90 days significant increases (P < 0.013) of MCP-1 graft uptake were seen at imaging and confirmed on scintillation gamma well counting studies in Lean (n = 5) and Fat (n = 12) animals, regardless of diet, 400 % and 40 %, above control values, respectively. MCP-1 uptake of native and grafted hearts correlated with increased numbers of perivascular macrophages (P < 0.02), as seen by immunostaining with an antibody specific for macrophages (ED 2). CONCLUSION: Radiolabeled MCP-1 can detect abnormally increased numbers of perivascular mononuclear cells in native and grafted hearts in prediabetic rats. MCP-1 may be useful in the screening of diabetic children for early atherosclerotic disease. PMID- 11727016 TI - Spiral CT scanning technique in the detection of aspiration of LEGO foreign bodies. AB - BACKGROUND: Radiolucent foreign bodies (FBs) such as plastic objects and toys remain difficult to identify on conventional radiographs of the neck and chest. Children may present with a variety of respiratory complaints, which may or may not be due to a FB. OBJECTIVE: To determine whether radiolucent FBs such as plastic LEGOs and peanuts can be seen in the tracheobronchial tree or esophagus using low-dose spiral CT, and, if visible, to determine the optimal CT imaging technique. MATERIALS AND METHODS: Multiple spiral sequences were performed while varying the CT parameters and the presence and location of FBs in either the trachea or the esophagus first on a neck phantom and then a cadaver. Sequences were rated by three radiologists blinded to the presence of a FB using a single scoring system. RESULTS: The LEGO was well visualized in the trachea by all three readers (both lung and soft-tissue windowing: combined sensitivity 89 %, combined specificity 89 %) and to a lesser extent in the esophagus (combined sensitivity 31 %, combined specificity 100 %). The peanut was not well visualized (combined sensitivity < 35 %). The optimal technique for visualizing the LEGO was 120 kV, 90 mA, 3-mm collimation, 0.75 s/revolution, and 2.0 pitch. This allowed for coverage of the cadaver tracheobronchial tree (approximately 11 cm) in about 18 s. Although statistical power was low for detecting significant differences, all three readers noted higher average confidence ratings with lung windowing among 18 LEGO-in-trachea scans. CONCLUSION: Rapid, low-dose spiral CT may be used to visualize LEGO FBs in the airway or esophagus. Peanuts were not well visualized. PMID- 11727017 TI - Digitizing pediatric chest radiographs: comparison of low-cost, commercial off the-shelf technologies. AB - OBJECTIVE: To compare low-cost, off-the-shelf technology for digitizing pediatric chest radiographs. MATERIALS AND METHODS: Forty pediatric chest radiographs (hard copy), each with a single abnormality, were digitized using a commercial film digitizer and two low-cost METHODS: a digital camera and a flatbed scanner. A stratified, randomized, block design was used where 20 readers evaluated 40 different images to determine the ability to accurately detect the abnormality. Readers then rated all 160 images (40 images x 4 methods) for conspicuity of the abnormality and overall image quality. RESULTS: Abnormalities were correctly identified on 82.3 % of hard copy images, 82.9 % of flatbed scanner images, 74.3 % of film digitizer images, and 69.7 % of digital camera images (p < 0.05) when compared to hard copy or flatbed scanner images. Lesion conspicuity was rated higher on hard copy (p < 0.05) than all digitized images. Conspicuity ratings were similar for flatbed scanner and film digitizer images, but lower in digital camera images (p < 0.05). For overall image quality, all were rated significantly different from each other (p < 0.05), with hard copy > flatbed scanner > film digitizer > digital camera images. CONCLUSION: A low-cost flatbed scanner yielded digital pediatric chest images which were significantly superior to digital camera images While flatbed scanner images were interpreted with the equivalent diagnostic accuracy of hard copy images, they were rated lower for image quality and lesion conspicuity. PMID- 11727018 TI - Isolated intrathoracic presentation of Mycobacterium avium complex in an immunocompetent child. AB - Mycobacterium avium complex (MAC) is a rare cause of isolated intrathoracic disease in the immunocompetent child; delays in diagnosis and treatment are common. The current case is an 11-month-old girl with symptoms, signs, and radiographic findings of bronchial obstruction. Comparison of this case to the literature revealed that there is a characteristic presentation of intrathoracic MAC infection in immunocompetent children (children aged 3 years or less, without exposure to Mycobacterium tuberculosis, presenting with clinical and radiographic evidence of bronchial obstruction) that should increase our index of suspicion for this diagnosis. This is particularly important since M. tuberculosis can present in a similar pattern. PMID- 11727019 TI - Analysis of the value of imaging as part of the follow-up of splenic injury in children. AB - OBJECTIVE: A recent article suggested that routine follow-up imaging is still frequently used in the conservative management of splenic trauma in children. The purpose of this study was to use decision analysis to assess the value of routine imaging as part of the long-term follow-up of splenic injury in children managed nonoperatively. METHODS: A literature review (1970-1999) on the management of blunt splenic trauma in children was performed. Data, including the use of follow up imaging and the occurrence of delayed splenic rupture and death, on those patients managed nonoperatively were collected. The data were used to construct a decision tree. A Poisson distribution was used to determine the risk of delayed splenic rupture. RESULTS: Information was extracted from 26 cohort studies. Nineteen of these studies were retrospective and six were prospective. One study had both retrospective and prospective arms. The study population consisted of 1,083 children. Of these patients, 920 (85 %) underwent routine follow-up imaging (US, CT, or scintigraphy). Follow-up imaging was either not performed or selectively performed in 163 patients (15 %). No cases of post-discharge splenic rupture or death were encountered in any of these groups. The maximum risk of delayed splenic rupture in the entire group was 0.3 % (0-3.7 cases). CONCLUSION: The risk of delayed splenic rupture following blunt injury in children is very low, and is apparently unaffected by imaging protocols. No deaths, even in cases of delayed presentation, were identified in our study. These findings do not support the use of routine follow-up imaging of children with blunt splenic trauma. PMID- 11727020 TI - Sonographic windsock sign of a duodenal web. AB - We report a neonate with a duodenal web demonstrating the windsock appearance on US. In neonates, duodenal web is rare and its windsock appearance is also rarely seen. The windsock sign of duodenal web has been a well-known finding on upper gastrointestinal series. The corresponding windsock appearance may be demonstrated on US. Duodenal web can, therefore, be accurately diagnosed by identifying the sonographic windsock sign even in neonates. PMID- 11727021 TI - Sacral development in anorectal malformations and in normal population. AB - BACKGROUND: The sacral ratio (SR) was proposed by Alberto Pena in 1995 as a reliable tool to evaluate sacral development in anorectal malformations (ARM). The SR is obtained by comparing sacrum size with fixed bony parameters of the pelvis. In previous studies, the average normal SR was calculated to be 0.74 for the anteroposterior view (APSR) and 0.77 for the lateral view (LSR). OBJECTIVES: (a) To evaluate the range of variability of the SR in a normal population with age as a possible variable factor. (b) To evaluate the reliability of the SR as a prognostic factor in patients with ARM. MATERIALS AND METHODS: We studied pelvic radiographs of 147 normal children in order to calculate a normal range of values for the SR. In 59 children with ARM, clinical parameters of faecal and urinary continence, spinal cord anomalies (as detected by MRI) and rectal innervation patterns (studied by enzymo-histochemistry) were correlated with SR values by Snedecor's 'F' test and Bravais-Pearson's 'r' correlation index. In 25 cases (15 children with ARM and 10 controls), multiple measurements of SR were obtained at different ages. RESULTS: Normal values for APSR ranged from 0.52 to 1.12, with an average of 0.74. Normal values for LSR ranged from 0.52 to 1.18, with an average of 0.75. Low SR values (below 0.52) correlated well with spinal cord anomalies and with unfavourable functional prognosis in children with ARM. APSR and LSR values increased with age in the normal population, while they decreased in patients with ARM. CONCLUSIONS: The SR has a wide range of normal values, and should be considered truly pathological when lower than 0.52, in either the AP or lateral views. Age has a significant influence on SR values. PMID- 11727022 TI - Infant shoulder sonography: technique, anatomy, and pathology. AB - BACKGROUND: In the infant, shoulder injury due to birth trauma and infection in joint and bone may be difficult to detect radiographically. Shoulder ultrasound provides a dynamic, noninvasive method of evaluation. OBJECTIVE: To develop a technique to evaluate the infant shoulder and to successfully demonstrate pathology. MATERIALS AND METHODS: Using a multiplanar, dynamic technique, 24 normal and 12 symptomatic infants (age range, 3 days to 9 months) were examined (49 shoulders). RESULTS: Sonographic findings were normal in 41 shoulders and abnormal in 8 shoulders. Abnormalities included subluxation, fracture, abnormal cartilage, soft-tissue mass, and inflammatory collections, including effusion. CONCLUSION: Ultrasound can be used successfully to evaluate the infant shoulder for instability, fracture, and infection. PMID- 11727023 TI - Imaging of malignant infantile osteopetrosis before and after bone marrow transplantation. AB - BACKGROUND: Malignant infantile osteopetrosis (MIOP) is a sclerosing bone disease caused by absence or defective function of osteoclasts. Since these are of haemopoietic origin, the disease can be cured by allogeneic stem-cell transplantation, but there are no detailed studies of radiological follow-up of these procedures. OBJECTIVE: To investigate the radiological findings at presentation and follow-up in children undergoing bone marrow transplantation (BMT) for MIOP. MATERIALS AND METHODS: Examination of the records and imaging studies of nine paediatric patients undergoing BMT for MIOP during 1988-2000. RESULTS: Presentation findings included characteristic features such as fractures, subperiosteal new bone formation and rachitic appearances. Five children engrafted successfully, allowing assessment of the nature and speed of resolution of radiological features after transplantation. CONCLUSIONS: Radiological improvement was apparent within 2 months of successful engraftment with almost complete resolution of abnormalities after 1 year. Studies in two children who are, respectively, 58 and 83 months post-transplant show complete resolution of all bone changes. PMID- 11727024 TI - A case of head and neck kaposiform hemangioendothelioma simulating a malignancy on imaging. AB - Kaposiform hemangioendothelioma (KH) is an endothelial-derived spindle cell neoplasm often associated with Kasabach-Merritt syndrome. Most cases arise in infancy and childhood and are soft-tissue tumors. The tumor displays an appearance between capillary hemangioma and Kaposi's sarcoma. We report a case of KH in a 1-year-old girl involving a mass that showed abnormal enhancement of soft tissue superficial to the right temporal bone with partial destruction of the temporal bone, the temporomandibular joint, mandibular condyle, and occipital bone. The physical finding of a discolored mass led clinicians to consider a hemangiomatous lesion, whereas the radiological picture suggested a more aggressive diagnosis of rhabdomyosarcoma and aggressive fibromatosis. PMID- 11727025 TI - Extramedullary hematopoiesis in a child with hereditary spherocytosis: an uncommon cause of an adrenal mass. AB - We report a case of extramedullary hematopoiesis presenting as an adrenal mass in a young male with hereditary spherocytosis. The unilateral adrenal mass was discovered during an abdominal ultrasound performed for jaundice. CT and MR imaging were subsequently performed, followed by an excisional biopsy at the time of splenectomy and cholecystectomy. Although extramedullary hematopoiesis is a rare cause of an adrenal mass, the diagnosis must be considered in any patient with a history of a congenital hemolytic disorder such as hereditary spherocytosis. In this regard, the morbidity of an unnecessary procedure may be avoided. PMID- 11727026 TI - Dynamic MR imaging of bladder haemangioma. AB - Haemangioma is a rare benign mesenchymal tumour of the bladder. We report a case of bladder haemangioma and describe the enhancement kinetics observed on MR images after gadolinium bolus administration, which is similar to that reported in liver haemangioma. PMID- 11727027 TI - Axial torsion of Meckel's diverticulum presenting as a pelvic mass. AB - Meckel's diverticulum is the most common congenital gastrointestinal anomaly. Axial torsion of the diverticulum is rare and may produce nonspecific abdominal signs and symptoms. We describe a case of torsion of a Meckel's diverticulum that was noted as a pelvic mass on CT images. PMID- 11727028 TI - Sickle cell anemia: the legacy of the patient (Walter Clement Noel), the interne (Ernest Irons), and the attending physician (James Herrick) and the facts of its discovery. PMID- 11727030 TI - The bank robber Willie Sutton, author of Sutton's Law: "go where the money is." Famous phrases in radiology. PMID- 11727031 TI - New dysplasia or achondrogenesis type 1B? The importance of histology and molecular biology in delineating skeletal dysplasias. PMID- 11727032 TI - Lumbar rhabdomyosarcoma in a patient with spina bifida. PMID- 11727033 TI - Transformation of Enterobacter gergoviae isolated from pink bollworm (Lepidoptera: Gelechiidae) gut with Bacillus thuringiensis toxin. AB - Production of molecules with toxic activity by genetically transformed symbiotic bacteria of pest insects may serve as a powerful approach to biological control. The symbiont, Enterobacter gergoviae, isolated from the gut of the pink bollworm (PBW), has been transformed to express Cyt1A, a cytolytic protein toxin lethal to mosquito and black fly larvae, as a model system. These transgenic bacteria might be used to spread genes encoding insecticidal proteins to populations of agricultural insects or as replacement for chemical insecticides such as malathion used in bait formulation to control specific insect pests, because of extreme public pressure against organophosphate pesticide spraying. PMID- 11727034 TI - Effect of oleic acid on Oenococcus oeni strains and Malolactic fermentation in wine. AB - A different capability to assimilate oleic acid from the culture medium has been demonstrated among malolactic Oenococcus oeni strains. Strains possessing higher percentages of oleic acid and its methylated derivative, dihydrosterculic acid, in their fatty acid profile showed higher cell viability and carried out a complete malolactic fermentation after their transfer into a wine lacking oleic acid. Wine supplementation with Tween 80 (polyoxyethylene-sorbitan-mono-oleate) enhanced cell survival of strains with lower capability to assimilate oleic acid and caused cell growth of strains with higher assimilative capacity, suggesting that oleic acid may act in wine as a survival factor for the former strains and as a growth factor for the latter strains. Practical consequences of these findings are also discussed. PMID- 11727035 TI - Characterization of anti-Listeria monocytogenes bacteriocins from Enterococcus faecalis, Enterococcus faecium, and Lactococcus lactis strains isolated from Raib, a Moroccan traditional fermented milk. AB - Seventy-four samples of raib, a Moroccan traditional fermented milk, were screened for their anti-Listeria monocytogenes activity. Nine lactic acid bacteria with antilisterial activity were isolated and identified as Lactococcus lactis [4], Enterococcus faecium [4], and E. faecalis [1]. Antibacterial spectra, determined against 45 target strains, led to the selection of four antibacterial producing strains, which were further characterized. Their anti-microbial agents, inactivated by one or more proteases, were designed as bacteriocins. Lactococcin R9/2 and R10/1 showed the broadest range of inhibitory action. Anti-bacterial spectra and physico-chemical properties suggest that these bacteriocins were similar to nisin. Enterocin R69 had a specificity of action against Listeria spp., whereas Enterocin R18 had a broad spectrum of activity. Lc. lactis R9/2 and E. faecalis R18 were able to coagulate sterilised UHT milk at 30 degrees C in 24 h and induced a 2 log reduction in L. monocytogenes ATCC 15313 population. PMID- 11727036 TI - Site-specific integration of the double-mutation glucose isomerase (GIG138PG247D) gene in Streptomyces lividans and its stable expression. AB - A recombinant expression plasmid pYH12, containing the double-mutation glucose isomerase (GIG138PG247D, GI2) coding gene and its natural regulatory sequence, was constructed for site-specific integration in Streptomyces. The resulting plasmid was introduced into Streptomyces lividans TK54 by protoplast transformation and two apramycin-resistance (AmR) transformants, designated GY2 and BY7, respectively, were obtained further based on enzyme assays. These results for polymerase chain reaction (PCR), Dot blot, and recovery of cloned fragments from the transformant chromosome indicated that the GI2 gene was integrated into the S. lividans chromosome by site-specific recombination, and which was further verified by Southern blot. We found that the free form of plasmid pYH12 co-existing with the integrated form was present in S. lividans. SDS-PAGE analysis showed that the GI2 gene was expressed in S. lividans. The intracellular GI2 specific activity was 1.15 U/mg. The stability of integrants demonstrated that the cloned GI2 gene was stably integrated and expressed even in the absence of selective pressure. PMID- 11727037 TI - A UV tolerant mutant of Bacillus thuringiensis subsp. kurstaki producing melanin. AB - A UV-resistant mutant (Bt-m) of Bacillus thuringiensis subsp. kurstaki, producing a dark brown pigment, identified as melanin, was studied. Bt-m had higher larvicidity against Heliothis armigera than its parent. Survival of Bt-m spores and their insecticidal activity to irradiation at 254 nm and 366 nm were higher than those of the parent. The only toxic polypeptide produced by Bt-m was Cry1Ac (130 kDa); it lost cry1Aa, cry2Aa, and cry2Ab. PMID- 11727038 TI - 4-Hydroxybenzoate uptake in Klebsiella planticola strain DSZ1 is driven by DeltapH. AB - Klebsiella planticola strain DSZ1 has the ability to degrade different aromatic compounds such as benzoate and organochlorinated as propachlor and alachlor. DSZ1 strain cells mineralised 4-hydroxybenzoate (4HBA) through a meta-cleavage pathway, yielding protocatechuate as dihydroxylated intermediate, with a specific rate of CO2 formation 0.12 x 10-6 (cpm/OD) h-1, and a rate of 4-HBA utilisation of 0.75 mmol h-1. Aerobically the 4HBA transport system is driven by gradient of protons (DeltapH), but is not ATP-driven. Under anaerobic conditions, the system can use the nitrate reduction as a final electron acceptor in respiration. A kinetic analysis of the 4HBA transport system revealed a Kt value of 16 microM with a Vmax value of 25 nmol/min.mg at pH 7. PMID- 11727039 TI - Carvacrol and thymol reduce swine waste odor and pathogens: stability of oils. AB - An incomplete anoxic fermentation of livestock waste results in offensive odor emissions. Antimicrobial additives may be useful in controlling odor emissions and pathogens. Natural antimicrobial compounds, carvacrol or thymol at 16.75 mM (2.5 g/l) completely inhibited the production of the offensive odor compounds, isobutyrate, valerate, isovalerate, and cresol, and significantly reduced other short-chain volatile fatty acids and gas emissions from swine waste. Fecal coliforms were reduced from 6.3 x 10(6) to 1.0 x 10(3) cells per ml 2 days after treatment with carvacrol (13.3 mM) and were not detectable within 14 days. Total culturable anaerobic bacteria were reduced from 12.4 x 10(10) to 7.2 x 10(8) cells per ml after 2 days and were suppressed below this level for 28 days. Lactate production was not prevalent in untreated swine waste indicating that the microbial populations differ from those in cattle waste. Carvacrol and thymol were stable in swine waste under anoxic conditions for 62 days with 90 to 95% of the additive being recovered in the waste solids. In conclusion, carvacrol and thymol are not metabolized in anoxic swine waste and they are potentially useful in controlling odor emissions and pathogens in swine waste. PMID- 11727040 TI - Multi-copy repression of Serratia marcescens nuclease expression by dinI. AB - The dinI homolog of S. marcescens was cloned from a plasmid library by virtue of its ability to inhibit nuclease expression from the S. marcescens nucA gene integrated in the genome of E. coli. The S. marcescens DinI protein is 68% identical to DinI of E. coli. It has a similar effect on other SOS regulated genes and likely exerts it effect on nuclease expression, which is most pronounced as the cells entered stationary phase, through inhibition of basal SOS expression. PMID- 11727041 TI - Nanoscale powders and formulations with biocidal activity toward spores and vegetative cells of bacillus species, viruses, and toxins. AB - Certain formulations of nanoscale powders possess antimicrobial properties. These formulations are made of simple, nontoxic metal oxides such as magnesium oxide (MgO) and calcium oxide (CaO, lime) in nanocrystalline form, carrying active forms of halogens, for example, MgO. Cl2 and MgO. Br2. When these ultrafine powders contact vegetative cells of Escherichia coli, Bacillus cereus, or Bacillus globigii, over 90% are killed within a few minutes. Likewise, spore forms of the Bacillus species are decontaminated within several hours. Dry contact with aflatoxins and contact with MS2 bacteriophage (surrogate of human enterovirus) in water also causes decontamination in minutes. PMID- 11727042 TI - Chemolithoautotrophic nitrifiers in the phyllosphere of a spruce ecosystem receiving high atmospheric nitrogen input. AB - Evidence is presented for the first time that chemolithoautotrophic ammonia oxidizers (CAO) and chemolithoautotrophic nitrite oxidizers (CNO) colonize in appreciable cell numbers the phyllosphere of spruce trees in a forest ecosystem exposed for decades to high levels of atmospheric nitrogen (The Hoglwald Forest, Bavaria, Germany). The results strongly indicate that both, CAO and CNO are predominantly located inside the spruce needles, most likely within the stomatal cavity. These results are further supported by field experiments of NH3 uptake into twigs on intact spruce trees in the presence and absence of 10 Pa acetylene, an inhibitor of the ammonia monooxygenase of CAO. It is clearly demonstrated for the first time that in situ uptake of NH3 from the atmosphere into spruce needles exposed to high levels of atmospheric N is not catalyzed exclusively by the tree, but is the result of combined activities of both, the spruce trees and the chemolithoautotrophic nitrifiers colonizing the needles. PMID- 11727043 TI - Proteome analysis of aniline-induced proteins in Acinetobacter lwoffii K24. AB - Acinetobacter lwoffii K24 is a soil bacterium that can use aniline as a sole carbon and nitrogen source (by beta-ketoadipate pathway genes (cat genes)) and has two copies of catABC gene separately located on the chromosome. In order to identify aniline-induced proteins, two-dimensional electrophoresis (2-DE) was applied to soluble protein fractions of A. lwoffii K24 cultured in aniline and succinate media. In the range of pH3-10, more than 370 spots were detected on the silver stained gels. Interestingly, more than 20 spots were selectively induced on aniline-cultured bacteria. Twenty-three protein spots of A. lwoffii K24 were analyzed by N-terminal microsequencing and internal microsequencing with in-gel digestion. Of 20 aniline induced protein spots, we identified six beta ketoadipate pathway genes, one subunit of amino group transfer (putative subunit of aniline oxygenase), malate dehydrogenase, putative ABC transporter, putative hydrolase, HHDD isomerase, and five unknown proteins. Especially in case of two catechol 1,2-dioxygenases (CDI1 and CDI2), more than three isotypes were detected on the 2D gel. This study showed that the proteome analysis of A. lwoffii K24 may be helpful for identification of genes induced by aniline and understanding of their function in the cell. PMID- 11727044 TI - Integration of the gene for carboxin resistance does not impact the Ustilago maydis-maize interaction. AB - A previous report indicated that insertion of the carboxin resistance (cbxR) gene into the Ustilago maydis genome impaired the pathogenic ability of the fungus towards Zea mays, the corn host. Because we had anecdotal evidence from work in our laboratory that this was not necessarily the case we decided to determine how general was the observation of reduced pathogenicity associated with cbxR. To accomplish this we tested the pathogenicity of several strains that had been transformed with the cbxR gene and compared them with non-transformed strains or strains transformed with the gene conferring hygromycin resistance which is a commonly used selectable marker in this fungus. Our results indicate that carboxin resistance does not significantly alter pathogenicity and is therefore a suitable marker for use in genetic analysis of U. maydis. PMID- 11727045 TI - Bacitracin: substantiation and elimination of contaminating proteolytic activity and use as an affinity chromatography ligand to purify a siderophore-degrading enzyme. AB - Bacitracin is a commercial general peptidase inhibitor that may be used to purify proteases. Significant protease contamination of a commercial bacitracin preparation was noted and four procedures were developed to overcome the contamination. Dialysis, gel-filtration chromatography, molecular weight cutoff filters, and heat inactivation were effective, resulting in the diminution or elimination of proteolysis while maintaining the inhibitory effect of bacitracin. Attachment of bacitracin to an affinity chromatography resin did not immobilize a siderophore-degrading enzyme, as has been noted with peptidases. It did, however, result in its partial purification from some of the contaminating proteins originally present. PMID- 11727046 TI - Indications for surgical treatment of funnel chest by chest radiograph. AB - Forty-seven children with funnel chest (FC) who underwent sternal elevation and 210 normal children were examined to determine the indications for surgical treatment using the vertebral index (VI) and frontosagittal index (FSI). In normal children VI gradually increased and FSI gradually decreased with age. Both indices changed significantly at 3 years of age. Although the VI of FC patients decreased significantly from 33.8 +/- 7.6 (n=40) to 24.4 +/- 3.9 (n=38) postoperatively (P < 0.0001), it was significantly larger than that of normal children over 3 years of age (20.2 +/- 2.2, n=150) (P < 0.0001), and although the FSI of FC patients increased significantly from 22.0 +/- 7.0 (n=40) to 34.5 +/- 6.5 (n=38) postoperatively (P < 0.0001), it was significantly smaller than that of normal children over 3 years of age (41.1 +/- 4.0, n=150) (P < 0.0001). Since many patients had a thin and flat chest despite excellent correction, their postoperative indices were not normal. There was a correlation between VI and FSI in normal children and a high degree of correlation between VI and FSI both before and after operation in FC patients. We conclude that a VI of more than 27 and/or a FSI of less than 29 are indications for surgical treatment based on the mean VI + 3SD and FSI - 3SD of normal children over 3 years of age. These values are almost equal to the mean VI - SD and FSI + SD of patients with physical, cosmetic, and/or psychological disturbances. However, it is not necessary to measure both indices simultaneously. Postoperative VI and FSI did not always reflect the degree of chest-wall depression in FC patients because of their flat chests. PMID- 11727047 TI - Infants with radiologic diagnosis of gastric volvulus: are they over-treated? AB - Gastric volvulus (GV) is a rare condition in infants. The aim of this study was to define the management strategies of infants with GV based on their clinical and radiologic features. The medical records of 13 infants with a radiologically confirmed diagnosis of GV were retrospectively reviewed. Patients were divided into two groups according to the type of treatment (surgical vs conservative). Abdominal radiographs and upper gastrointestinal contrast studies allowed an unequivocal diagnosis in both groups. Group 1 included 3 infants with acute GV and 2 with chronic, intermittent secondary GV. Three patients had associated diaphragmatic defects, 1 had an ileocolic intussusception, and 1 had hypertrophic pyloric stenosis. The main presenting symptoms were vomiting, dehydration, respiratory distress, and abdominal pain and distention in acute cases and vomiting and failure to thrive in chronic cases. A laparotomy was required in all 5 infants with no recurrence of symptoms. Group 2 included 8 infants with idiopathic chronic GV, who were managed nonoperatively with gradual improvement of symptoms over 12 months. Based on our study, we conclude that: (1) laparotomy can be reserved for patients with either acute or chronic secondary GV; (2) conservative treatment is both safe and effective in infants with chronic idiopathic GV; and (3) routine gastropexy for all patients with a radiologic diagnosis of GV appears to be overtreatment. PMID- 11727048 TI - Clinical significance of c-kit expression in biliary atresia. AB - The aim of the present study was to examine the clinical significance of c-kit expression in biliary atresia (BA) using formalin-fixed, paraffin-embedded sections from 21 patients with BA. Patients were divided into group I (n = 8) with good liver function; group II (n = 8) with moderate liver dysfunction; and group III (n = 5) with severe liver dysfunction. Choledochal cysts (CDC, n = 5) and normal liver samples (NL, n = 4) served as controls. The results were analyzed and compared among the groups. Most c-kit+ cells were present in the portal tracts, and their numbers in BA were significantly higher than in the controls (11.12 +/- 1.64 vs 2.15 +/- 0.15 [mean +/- standard error], P = 0.02, BA vs CDC; 11.12 +/- 1.64 vs 1.66 +/- 0.52, P = 0.03, BA vs NL). Clinical correlation revealed a significantly higher number of c-kit+ cells in group III versus group I (18.10 +/- 3.62 vs 8.86 +/- 2.51, P = 0.02). These results suggest that c-kit overexpression is associated with an adverse clinical outcome in BA. PMID- 11727049 TI - Bone density and 25-hydroxyvitamin D level in extrahepatic biliary atresia. AB - Biliary atresia (BA) represents a common cholestatic affliction of the gastrointestinal tract affecting infants and children. The objective of the present study was to evaluate 42 patients (20 with and 22 without jaundice) diagnosed with extrahepatic BA for bone mineral content and serum 25 hydroxyvitamin D (HVD) levels. Physical examination and anthropometric nutritional assessment were performed. The investigation included liver function tests and serum calcium (Ca), phosphate (P), magnesium (Mg), and 25-HVD levels. Dual-energy X-ray absorptiometry was used to measure the bone mineral density (BMD) of the lumbar spine (L(1)-L(4)). Our results showed that 16 jaundiced patients (80%) and only 3 nonjaundiced patients (13.6%) showed osteoporosis (P< 0.05). All patients had normal serum Ca and P levels. Only 1 nonjaundiced patient had a low serum Mg level. Serum 25-HVD levels (mean +/- SD) were 20.71 +/- 8.24, 16.12 +/- 4.3, and 9.18 +/- 5.84 ng/ml, respectively, in subjects with normal bone density (n=7), osteopenia (n=3), and osteoporosis (n=11). Bone disease represents a well-known complication among long-term survivors of BA. To date, the pathogenesis has remained unexplained. Since, as demonstrated in the present study, jaundiced patients develop osteoporosis more frequently than nonjaundiced patients, hyperbilirubinemia may have an influence. Bone-mineral deficiency can be detected earlier by means of BMD measurement (non-invasive method) than by measuring serum Ca, P, and Mg levels in these patients. PMID- 11727050 TI - In-vitro uptake of radioactive lipiodol I-131 and I-125 by hepatoblastoma: implications for targeted radiotherapy. AB - Lipiodol, a stable iodine addition product of fatty ethyl esters derived from poppyseed oil, has been used as a vehicle for targeted cytotoxic or radiotherapeutic treatment in adults with unresectable hepatocellular carcinoma and in a few children with advanced hepatoblastoma (HB). Prolonged retention of lipiodol by the cancer cells might enable more effective targeted therapy to advanced, multifocal, or metastatic HB. To investigate the uptake and cytotoxic efficacy of beta and Auger electron-emitting radioconjugates on HB cells, monolayers of HB (C3a, Hep Tow1) and normal human hepatocyte cell lines were exposed to lipiodol131 (L131), lipiodol125 (L125), or a cocktail of both isotopes. Uptake of radioactivity was assessed autoradiographically using phosphoimages and the cytotoxicity assessed by trypan blue exclusion and clonogenic assay. The uptake of the different radioconjugates was identical in both HB cell lines. The "cocktail" of both radioactive lipiodols caused the greatest cytotoxicity to HB C3a, and HepTow1. L125 alone had a similar level of uptake and cytotoxicity as L131. Radioactive iodine alone did not show any cytotoxicity on any of the liver cell lines in culture for up to 72 h. These experimental results provide support for a clinical strategy of combinations ("cocktails") of radioconjugates in targeted radiotherapy in patients with HB. PMID- 11727051 TI - Solid-cystic papillary tumor of the pancreas in children. AB - Solid-cystic papillary tumor (SCPT) of the pancreas is a rare neoplasm in children. The current study attempted to estimate the incidence and possible pathological origin of the tumor. Clinicopathologic features of the children under the age of 16 years with pancreatic tumors managed in a single institution between January 1993 and November 1999 were reviewed. Representative blocks of SCPT specimens were immunostained for neuron-specific enolase (NSE) chromogrannin, synaptophysin, insulin, glucagon, somatostatin, and gastrin. There were three SCPTs, two pancreatic endocrine tumors, and one acinar cell carcinoma during the study period. The estimated yearly incidence in the referral area of 2 million population is about 0.01 pediatric SCPT per year per 100,000 population. The children underwent surgical removal of the tumors. Postoperatively, they were followed up for 6 months to 4 years and were well. Immunostaining for NSE, chromogranin, synaptophysin, insulin, somatostatin, and gastrin was negative in all cases. SCPT may thus be the most common pancreatic tumor in the Asian pediatric population. The pathological origin of the tumor remains unclear and requires further investigations. PMID- 11727053 TI - Detection of bacterial translocation during intestinal distension in rats using the polymerase chain reaction. AB - To investigate whether distension causes bacterial translocation (BT), a rat model reported earlier by us was used and to detect the presence of bacterial DNA in blood by polymerase chain reaction (PCR) assay, the most sensitive detection method to date. In 4 groups of 4-week-old Wistar-albino rats a total of 15 animals each were included. In the 1st group (distension+gavage), 1010 Escherichia coli colonies were given via gavage and distension was carried out by rectal air inoculation. In the 2nd group (gavage), animals were inoculated with E. coli and no distension was induced. The 3rd group (distension) were only distended and no bacteria were inoculated. The control group were neither distended nor inoculated with E. coli. Blood samples were collected 3 h after manipulations and both blood cultures and PCR assays were performed. According to the PCR results BT was evident in 80% of group 1, 20% of group 2, and 33% of group 3 animals. BT was not determined in the control group. Significantly low percentages of positivity were observed in blood cultures in all groups (P < 0.05). These results confirm reports that BT occurs in the presence of distension and that PCR is a superior way of determining BT. Thus, it would be advisable to utilize PCR technology in cases where the possibility of distension exists, as early intervention might be useful before any severe clinical pathology (sepsis, multiple-organ-system failure) becomes evident. PMID- 11727052 TI - Internal drainage of pancreatic pseudocysts in children using an endoscopically placed stent. AB - Persistent pseudocysts, which are rare in children, have traditionally been managed by open surgery. We describe two children who presented with large, established pseudocysts of the pancreas. They were successfully treated in the short term by minimally-invasive stent placement to create an internal cyst gastric communication. This approach merits comparison with open surgical techniques. PMID- 11727054 TI - Typhoid ileal perforation in Nigerian children: an analysis of 106 operative cases. AB - Typhoid ileal perforation (TP) is a major problem in developing countries and carries a high mortality. The purpose of this retrospective study from Nigeria was to review the outcome in children less than 15 years of age who underwent surgery for TP from 1984 to 1999. Demographic data, clinical features, results of investigations, findings at surgery, postoperative course, and complications were recorded. There were 55 boys and 51 girls. The median age at presentation was 10 years (range 3-14). The surgically confirmed perforation rate was 11.0%. The clinical features in children older than 5 years were similar to already documented patterns in the literature. In children less than 5 years old the predominant symptoms were fever and vomiting only. Therefore, in the very young a high index of suspicion is required to avoid delay in diagnosis. Ninety eight patients (92.5%) had simple double-layer closure of the perforation. The mean hospital stay among survivors was 23.6 +/- 18.8 days. The commonest postoperative complications were wound infection and enterocutaneous fistula. The overall mortality was 23.8%, increasing to 50% in children aged less than 5 years, although the difference was not statistically significant (P > 0.05). To improve survival in TP, attention should be focused on perioperative resuscitation and early intervention. The provision of potable water, adequate sanitation, and active immunisation are means to eradicate the disease. PMID- 11727055 TI - Colonic atresia: surgical management and outcome. AB - Colonic atresia (CA) is a very rare cause of intestinal obstruction, and little information has been available about the management and predictors of outcome. A retrospective clinical trial was performed to delineate the clinical characteristics of CA with special emphasis on surgical treatment and factors affecting outcome. Children with CA who were treated in our department between 1977 and 1998 were reviewed: 14 boys and 4 girls aged 1 day to 5 months. All but 2 referred patients and 1 with prenatal diagnosis presented with intestinal obstruction. Plain abdominal X-ray films showed findings of intestinal obstruction in 14 cases; a barium enema demonstrated a distal atretic segment and microcolon in 4. The types of atresia were IIIa (n=9), I (n=6), and II (n=3). Type IIIa atresias were located proximal to the splenic flexure (n=8) and in the sigmoid colon (n=1), type I atresias were encountered throughout the colon; and all type II atresias were proximal to the hepatic flexure. Associated anomalies were multiple small-intestinal atresias (MSIA) (n=4), gastroschisis (GS) (n=2), pyloric atresia (n=1), Hirschsprung's disease (n=1), and complex urologic abnormalities (n=1). The initial management was an enterostomy in 15 patients (83%), including 2 referred and 2 with GS, and primary anastomosis in the remaining 3 (17%). Secondary procedures were the Santulli operation (n=2), colostomy closure and recolostomy followed by a Swenson operation (n=1), sacroabdominoperineal pull-through (n=1), and colostomy closure (n=1). Leakage was encountered in all primarily anastomosed patients. The overall mortality was 61%. Deaths occurred in patients with associated major anomalies (GS 2, MSIA 3, pyloric atresia 1) (55%) and in 3 patients who were initially managed by primary anastomosis (27%). Two additional patients died of sudden infant death syndrome (18%). Type I atresia was more common than in previously reported series and was associated with proximal multiple atresias. The initial management of CA should be prompt decompression of the intestine by an ostomy procedure, preferably end- or double-barrel. The type of surgery (primary anastomosis without prior colostomy) and associated abnormalities are the major determinants of poor outcome. PMID- 11727056 TI - Sigmoid volvulus and ileosigmoid knotting in children. AB - Six children with acute intestinal obstruction from sigmoid volvulus (SV) (n=2) and ileosigmoid knotting (ISK) (n=4) are reported. The median age at presentation was 4.5 years (range 2 weeks-15 years). Abdominal pain, distention, vomiting, and constipation were the main features. Two patients with ISK had bowel gangrene. In three children there was no identifiable cause; two had a narrow attachment of the sigmoid mesocolon with redundant colon and one had adhesive bands. Treatment was by resection and colostomy in five cases and derotation of the torted colon in one. One child with SV died following a wound infection. There was no recurrence. SV and ISK are uncommon in children. There are usually no features specific for these conditions, and the diagnosis is established at laparotomy. The prognosis is good when there is aggressive resuscitation and prompt surgery. PMID- 11727057 TI - Single-system ectopic ureter: a 15-year review. AB - Ectopic ureters present in childhood with symptoms related to an abnormal site or structure (refluxing, obstructed) of the ureteric orifice. The majority drain duplex kidneys. The diagnosis is relatively easy if the poles are functioning or hydronephrotic. Associated malformations are rarely seen and the results of surgery are gratifying. If an ectopic ureter drains a single kidney, it is called a single-system ectopic ureter (SSEU). We reviewed a 15-year experience (1980 1995) with 127 ectopic ureters from our hospital:11 SSEUs in ten consecutive children were managed during this period. Our data lead us to believe that SSEUs are a special subset of ectopic ureters. Diagnosis is often delayed because the ectopic ureter may be associated with a single small, dysplastic, poorly functioning, non-visualised kidney and the child may be thought to have a contralateral normal "solitary kidney". Associated systemic malformations are common. Residual symptoms of wetting may persist in the early postoperative period. A high degree of suspicion must be maintained for this entity when a child presents with urinary symptoms of wetting or recurrent infection and a "solitary kidney". Early endoscopic examination of the genitourinary tract will clinch the diagnosis in the majority of cases. Residual symptoms of wetting in the postoperative period generally resolve with passage of time. PMID- 11727058 TI - H-type tracheo-oesophageal fistula: appearance on three-dimensional computed tomography and virtual bronchoscopy. AB - H-type tracheo-oesophageal fistula (TOF) is a rare type of fistula without oesophageal atresia. The symptoms are usually present at birth, but the diagnosis is often delayed. Traditionally, contrast oesophagography and/or fiberoptic bronchoscopy are used to confirm the diagnosis. We describe the imaging features of a case of H-type TOF seen on three-dimensional computed tomography and virtual bronchoscopy and the usefulness of these techniques in pre-operative evaluation. PMID- 11727059 TI - Neonatal acute appendicitis: a strangulated appendix in an incarcerated inguinal hernia. AB - A 4-day-old infant with acute appendicitis (AA) due to incarceration of a right inguinal hernia is presented. Although appendicitis is the most common condition requiring abdominal surgery, the reported occurrence of AA subsequent to neonatal hernia incarceration is exceptionally rare. PMID- 11727060 TI - Urgent distal splenorenal shunt in low-body weight patients. AB - The authors report two patients with extrahepatic portal hypertension and repeated massive bleeding from esophageal varices who underwent urgent distal splenorenal shunting (DSRS) after having proved refractory to medical treatment, endoscopic sclerotherapy, and ligation of esophageal varices. Their ages and weights were 18 months/10 kg, and 11 months/6.4 kg, respectively. The splenic veins were 6 and 4 mm in diameter. During follow-up of 4 and 3 years, respectively, the shunts have remained patent in both patients as demonstrated by Doppler Ultrasound. Gastrointestinal hemorrhage has not recurred, splenomegaly regressed, and platelet and white blood cell counts increased gradually. Neither patient developed a significant encephalopathy, and liver function tests showed no significant changes throughout the observed period. The authors considered the shunting feasible despite the relatively narrow splenic veins. In both patients the anatomic position of the splenic vein--more caudal to the posterior wall of the pancreas--facilitated its isolation. In the smaller infant, the inferior mesenteric vein was sutured to gain a satisfactory segment of splenic vein. The successful use of a DSRS to control actively bleeding varices in a child weighing 6.4 kg has not been previously reported. PMID- 11727061 TI - A rare anorectal malformation: a very large H-type fistula. AB - A very large H-type rectovaginal fistula was observed in a 9-year-old girl. The diameter of the abnormal opening was about 15 mm. The fistulous tract originated from the stenotic segment 1 cm above the pectinate line of the anus and was connected to the back wall of the vagina. The patient underwent posterior sagittal transrectal surgery with a protecting colostomy. At present, she has good bowel function and sphincter control without recurrence of the fistula. PMID- 11727062 TI - Paratesticular pilomatricoma: a new location. AB - The first case of pediatric paratesticular pilomatricoma is reported. Differential diagnosis with other more common lesions in such a site is the main issue. Conservative surgery is the treatment of choice. Follow-up is recommended, since most pilomatricomata are benign, but rarely malignant transformation may occur. PMID- 11727063 TI - Spermatic cord metastasis from a medulloblastoma. AB - A 4-year-old boy presented with metastases in the spermatic cord and ribs 2 years after treatment for a primary medulloblastoma (MB). The testis was free of tumor. A MB presenting with a metastasis to the spermatic cord has not been reported to date. The role of a ventriculo-peritoneal shunt in promoting such spread should be considered in planning management. PMID- 11727064 TI - Air-gun pellet injuries to the head and neck in children. AB - Air-gun pellet injuries to the head and neck are seldom reported in pediatric practice, although they typically occur in children. The adult skeleton stops these projectiles, but they can easily transverse the thin bones of children. If unnoticed, these apparently trivial injuries may have catastrophic consequences. We report three children who sustained a central nervous system injury resulting from a shot by a compressed-air gun. The true nature and extent of the lesion in two infants was established only by neuroradiological investigations. We also briefly review the management and prevention of this type of injury. PMID- 11727065 TI - Intestinal atresia: an end-to-end linear anastomotic technique. AB - In a series of 19 neonates with small-bowel atresia, 16 were treated by end-to end linear anastomosis (ELA) without resection and 3 by resection anastomosis (RA). Seven atresias were jejunal, 11 were ileal, and 1 jejunoileal; 3 cases were type II, 12 type IIIa, 3 type IIIb, and 1 type IV. There were 4 deaths, 1 after ELA and 3 after RA. The overall mortality decreased from 68 to 20.80% and for linear anastomosis to 6.25% presumably, because the intestinal contents are propelled along the lumen in a linear fashion and not at an angle as in end-to back anastomosis, avoiding shearing of the suture line. The additional plicating sutures reduce the radius and increase the propelling force. We recommend this technique because it is based on sound principles of physics and preserves the entire available length of intestine. PMID- 11727066 TI - Elastic bandaging facilitates primary closure of large ventral hernias due to giant omphaloceles. AB - Large ventral hernias (VH) following conservative treatment of a giant omphalocele are a major challenge for pediatric surgeons. Although primary closure is accepted as the ideal procedure, these defects often require multi staged operations, prosthetic support, and postoperative mechanical ventilation. Between 1990 and 2000, 34 patients with omphaloceles were admitted to our unit and 6 of them (17.6%) who had giant omphaloceles were treated conservatively. In the same period, a 5-year-old girl was admitted with a huge VH following application of subcutaneous tissue expanders in another center. In these 7 patients (6 girls, 1 boy, average age 20 months) we used preoperative elastic bandaging for facilitating primary closure. In all cases primary closure was successful following 3 to 6 weeks of bandaging. We did not use prosthetic material or postoperative mechanical ventilation in any case. Follow-up periods ranged from 2 to 10 years, and there were no recurrences or other problems. Out limited experience reveals that preoperative elastic bandaging is a safe and effective procedure for facilitating primary closure of the large VHs. PMID- 11727067 TI - Zhang's ring-clamp modification of Duhamel's procedure: 30 years experience. AB - To evaluate the efficacy of the ring-clamp procedure (RCP) in the treatment of Hirschsprung's disease (HD) 435 RCPs were compared to 54 Kocher procedures performed between 1960 and 1990. The data were analyzed by chi-square test. The RCP resulted in much less blind-pouch syndrome (7%) compared to the Kocher's procedure (40%), and represents a valid alternative in the treatment of HD in children. PMID- 11727068 TI - A new colonic lavage method for infants with Hirschsprung's disease. PMID- 11727069 TI - Wide-gap esophageal atresia. PMID- 11727070 TI - The digital surgeon. PMID- 11727071 TI - The need for training opportunities in advanced laparoscopic surgery. AB - BACKGROUND: There is controversy regarding the amount of training necessary to safely perform advanced laparoscopic surgery. General surgical residency often provides only a low volume of advanced laparoscopic cases and there is growing interest in nonaccredited fellowships focused on laparoscopic surgery. OBJECTIVE: To assess surgical residents' perception of the need for training in advanced laparoscopic surgery in addition to that provided in a standard general surgical residency. METHODS: A 15-item questionnaire was mailed to 985 physicians who either were Society of American Gastrointestinal Endoscopic Surgeons (SAGES) candidate members or had attended a SAGES resident course in 1998 or 1999. For the purposes of the survey, laparoscopic Nissen fundoplication, laparoscopic herniorrhaphy, laparoscopic splenectomy, and laparoscopic colectomy were chosen as advanced procedures. RESULTS: Of the 85 responses obtained, 81% were from respondents who were at the postgraduate fourth-year (PG4) level or higher. Furthermore, 58% of the respondents had taken a course in advanced laparoscopic surgery outside their residency program. The respondents believed that to perform the procedures safely and with confidence on entering practice, they needed to do at least eight each of the selected laparoscopic procedures. As reported, 45% of the respondents had performed three or fewer laparoscopic hernias; 60% had performed three or fewer laparoscopic Nissen fundoplications; 81% had performed three or fewer laparoscopic colectomies; and 86% had performed three or fewer splenectomies. Only 32% of the residents expected to perform more than 10 laparoscopic Nissen fundoplications, only 10% expected to perform more than 10 colectomies, and only 4% expected to perform more than 10 splenectomies before completing their residency. Many respondents (65%) said they would pursue an additional year of advanced laparoscopic training if it were available. In programs unaffiliated with a fellowship in advanced laparoscopic surgery, 65% of the residents were concerned that such a fellowship would interfere with residency training in laparoscopic surgery. In comparison, only 24% of the residents in programs affiliated with a fellowship in advanced laparoscopic surgery believed that the fellowship interfered with their training, whereas 47% of the residents in programs affiliated with a fellowship in advanced laparoscopic surgery thought that the fellowship had no impact on their training. CONCLUSIONS: Residents clearly perceive a need for additional training in advanced laparoscopic surgery. Residents from programs without a laparoscopic fellowship are concerned about a negative impact on their experience from a laparoscopic fellow, but residents from programs with a laparoscopic fellowship are neutral about the impact of a fellow. PMID- 11727072 TI - The role of the basic surgical skills course in the acquisition and retention of laparoscopic skill. AB - BACKGROUND: This study assesses the transfer of laparoscopic skills to a group of Basic Surgical Trainees (BST) attending the Basic Surgical Skills (BSS) course. METHODS: The virtual reality simulator MIST-VR was used to assess 13 trainees before and after the course and again 3 weeks and 3 months later. Analysis of kinematic data using the Imperial College Surgical Assessment Device gave measures of distance traveled, distance efficiency ratio, time taken, number of errors made, and number of movements made in completing a virtual laparoscopic task. The performance of the group was compared to a control group who underwent no training. RESULTS: All parameters improved significantly after the course, with the exception of distance traveled by the instruments. All outcome measures were significantly improved at 3 weeks. The control group showed a nonsignificant trend toward improvement in all parameters. CONCLUSIONS: The Basic Surgical Skills course produces quantifiable improvements in laparoscopic skill that are measurable by MIST-VR. There is a learning effect associated with using MIST-VR alone. PMID- 11727073 TI - Skill transfer from virtual reality to a real laparoscopic task. AB - BACKGROUND: To validate the usefulness of virtual reality surgical simulators, we investigated the transfer of skills achieved by their use to real tasks. METHODS: Thirty medical students underwent a pretest using a real laparoscopic trainer. They were then randomized to the following three groups: group I received no training; group II received training using the Minimal Invasive Surgical Trainer in Virtual Reality (MIST-VR); and group III received training using conventional training exercises. Each group then underwent a posttest. Using the Imperial College Surgical Assessment Device (ICSAD), scores were generated for time taken, distance traveled, number of movements made, and speed of instrument movement. RESULTS: Significant changes between the MIST-VR group (group II) and the conventionally trained group (group III), were observed in the speed of movement of the left hand and the numbers of movements taken by each hand, when compared to the untrained group (group I). CONCLUSION: The training of novices using MIST VR yields quantifiable changes in skill that are transferable to a simple real task and are similar to the results achieved with conventional training. PMID- 11727074 TI - Virtual reality training leads to faster adaptation to the novel psychomotor restrictions encountered by laparoscopic surgeons. AB - BACKGROUND: The fulcrum effect of the body wall on instrument handling poses a major obstacle to the mastery of instrument coordination for junior laparoscopic surgeons. This study evaluated three types of laparoscopic simulator training to assess their ability to promote the user's adaptation to the fulcrum effect. METHODS: Thirty-two participants with no previous experience in laparoscopic surgery were randomly assigned to one of four groups representing different training conditions. One group was assigned to use a virtual reality simulator (MIST VR); two others were given a laparoscopic Z or U maze-tracking task. The control group received no training. Subjects were asked to perform a 2-min laparoscopic cutting task under normal laparoscopic imaging conditions first before and then after training. RESULTS: In the test trial, subjects who trained on MIST VR made significantly more correct incisions (p < 0.0001) and fewer incorrect incisions (p < 0.0001). CONCLUSION: Training on a virtual reality simulator such as MIST VR helps laparoscopic novices adapt to the fulcrum effect faster. PMID- 11727075 TI - A computer-based laparoscopic skills assessment device differentiates experienced from novice laparoscopic surgeons. AB - BACKGROUND: The acquisition of laparoscopic skills is difficult to assess. Previously, through the use of a computer-based skills assessment device (SAD), we have shown that assessments of the time needed to perform a task and graded observations of task performance overestimate the speed at which laparoscopic skills can be acquired. The aim of this study was to test the ability of a laparoscopic SAD to differentiate novices from experienced laparoscopic surgeons and thereby further validate its use in surgical education. METHODS: Using a laparoscopic simulator platform integrated with task simulation and data collection software (MIST VR), we tested both experienced and novice laparoscopists. Each group performed three tasks with a minimum of seven repetitions. The tasks consisted of target acquisition, target traversal, and target manipulation with diathermy. Within each task, time (T), errors (E), and economy of movement for each hand (EML, EMR) were assessed. Results were reported as mean +/- SEM, and comparisons were made using an independent samples t-test. RESULTS: For the target acquisition task, the expert group performed the task faster than the novice group (5.5 +/- 0.24 vs 7.6 +/- 0.40 sec, p < 0.05); the experts also made fewer errors (0.5 +/- 0.10 vs 0.8 +/- 0.11 errors, p < 0.05), and achieved both a better EML (1.8 +/- 0.6 vs 2.3 +/- 0.9, p < 0.05) and a better EMR (2.0 +/- 0.1 vs 2.9 +/- 0.21, p < 0.05). In the target traversal task, the experts made fewer errors than the novices (2.2 +/- 0.25 vs 4.6 +/- 0.38 errors, p < 0.05). For the manipulation and diathermy tasks, the expert group completed the task faster (30.8 +/- 1.5 vs 39 +/- 1.5 sec, p < 0.05), made fewer errors (5.3 +/- 0.59 vs 8.1 +/- 0.63 errors, p < 0.05), and had a better EML (6.0 +/- 0.37 vs 7.2 +/- 0.45, p < 0.05) and EMR (4.3 +/- 0.23 vs 5.8 +/- 0.36, p < 0.05) than the novices. CONCLUSION: A laparoscopic skills assessment device can discern levels of laparoscopic manipulative skill. This type of device can be used in surgical training to make objective assessments of the acquisition of laparoscopic skills. PMID- 11727076 TI - Laparoscopically assisted subtotal colectomy for slow-transit constipation. AB - BACKGROUND: When medical therapy fails, slow-transit constipation (STC), a condition seen almost exclusively in women, can be treated surgically. The aim of this study was to describe our results with laparoscopically assisted subtotal colectomy (LASC) for STC. METHODS: Over a 22-month period, four female patients underwent LASC with ileorectal anastomosis for STC at our hospital. The preoperative workup included clinical assessment, evacuation proctography, rectoanal manometry, and colonic transit measurement. Mobilization of the whole colon and ligation and division of colonic mesenteric vessels with preservation of the omentum were achieved laparoscopically. Through a small Pfannenstiel incision, the colon was removed from the abdomen, leaving a 15-cm rectal stump in situ. The rectal stump was fully mobilized posteriorly, a side-to-end ileorectal anastomosis was fashioned, and the rectal stump was fixed onto the presacral fascia with sutures. RESULTS: The operating time ranged from 150 to 260 min and blood loss was minimal. There were no conversions to open surgery. The postoperative course was uneventful in all patients but one, a 47-year old woman who had transient severe abdominal distension, pain, and frequent small-volume bowel movements. All patients were discharged by the 4th to 10th postoperative day. At 9-month follow-up, all of our patients had normal anorectal function with two to four solid bowel movements per day. CONCLUSION: Although it is a technically demanding procedure, laparoscopically assisted colectomy for slow transit constipation, can be achieved safely. PMID- 11727077 TI - Gasless laparoscopy-assisted colorectal surgery. AB - BACKGROUND: Laparoscopy has gained wide acceptance as a treatment modality in a variety of colonic and rectal disorders. Currently, most laparoscopic procedures are performed using a carbon dioxide (CO2) pneumoperitoneum, which can lead to cardiopulmonary loading and subsequent complications. The object of this study was to assess the feasibility of gasless laparoscopy-assisted colorectal surgery (GLACS) as an alternative method. METHODS: Patients with benign colonic lesions were enrolled in the study. The operative field was exposed with a subcutaneous wire lifting system. A small incision, ~5 cm in length, was made early in the operation. The surgeon operated through the trocar ports and this incision using both laparoscopic and conventional instruments. The cardiopulmonary responses of the patients were monitored continuously during the operation. RESULTS: Fifteen consecutive patients underwent GLACS. In two patients (13.3%), conversion to open surgery was necessary. The exposure and ease of the procedure were acceptable. However, when the patients were stratified into hemicolectomy and sigmoidectomy groups, GLACS scored more favorably in the sigmoidectomy group. There were no operative deaths. One minor complication developed postoperatively. All of the patients recovered uneventfully, with return of bowel function in 2.8 +/- 0.1 days. The mean postoperative hospital stay was 6.4 +/- 0.4 days. The cardiac and pulmonary status of the patients remained stable during the operation. CONCLUSION: Gasless laparoscopy-assisted colorectal surgery is technically feasible; thus, it provides an alternative means for the performance of minimal access surgery. PMID- 11727078 TI - Laparoscopic-assisted total mesorectal excision and colonic J pouch reconstruction in the treatment of rectal cancer. AB - BACKGROUND: Total mesorectal excision (TME) and colonic J pouch reconstruction has been widely practiced for mid- or low-rectal cancer. However, the laparoscopic version of TME has never been described. METHODS: Five patients suffering from newly diagnosed mid- to low-rectal cancer were seen between March and July 1999. These five patients were selected for laparoscopic TME and colonic J pouch reconstruction because preoperative investigations revealed resectable tumor without extrarectal disease. RESULTS: There were three men and two women with a mean age of 61 years. The average body weight was 69 kg (range, 57-80). None of the patients had had previous abdominal operations. In all five patients, the tumor was located within 9 cm from anal verge. The average size of the main incision was 5 cm. All patients had a covering ileostomy at the end of the procedure. The mean operating time was 208 min; average blood loss was 158 ml; and mean hospital stay was 10.6 days. Three patients had Dukes' B disease and two had Dukes' C disease. The resection margins (proximal, circumferential, and distal) were all clear. There were no deaths or major complications. Two patients suffered from transient urinary retention. After ileostomy closure, the median frequency of bowel motion was twice per day at 6-month follow-up. Neither incontinence nor nocturnal soiling was reported. CONCLUSION: To the best of our knowledge, this is the first published series of such an operation. With good patient selection, laparoscopic-assisted TME and colonic J pouch-anal anastomosis is safe and feasible. PMID- 11727079 TI - Outcomes of laparoscopic herniorrhaphy without fixation of mesh to the abdominal wall. AB - BACKGROUND: Recently there has been interest in performing laparoscopic herniorrhaphies without the use of staples or tacks to fix the mesh. Although mesh fixation has been linked to an increased incidence of nerve injury and involves increased operative costs, many surgeons feel that fixation is necessary to reduce the risk of hernia recurrence. This study evaluates the outcomes of laparoscopic herniorrhapies performed with and without mesh fixation at our institution. METHODS: We retrospectively evaluated our last 172 laparoscopic herniorrhaphies, which span a period of conversion from staple fixation to nonfixation of total extraperitoneal herniorrhaphies using systematic chart review and follow-up self-administered questionnaires. The outcomes assessed were the incidences of postoperative neuralgia and hernia recurrence. Adjustment for important prognostic factors was achieved using Cox regression for estimating the risk of recurrence, and multiple logistic regression for estimating the risk of neuropathic complications. RESULTS: Of 172 laparoscopic herniorrhaphies performed in 129 patients since July 1993, 105 were accomplished without mesh fixation, and 67 were performed with fixation of mesh to the abdominal wall. There were no significant differences in demographics between the two groups. A trend toward a higher incidence of neuropathic complications was observed in the mesh-fixation group (risk ratio [RR], 2.2; 95% CI, 0.5-10). A nonsignificant increased risk of hernia recurrence with fixation of mesh was observed (4.2 vs 1.6 per 100 hernia years at risk; RR, 2.3; 95% CI, 0.4-13.10), but this finding may be associated with a selection bias with regard to giant hernia defects. CONCLUSIONS: Our data suggest that mesh fixation to the abdominal wall may be avoided in total extraperitoneal repairs without increasing the risk of hernia recurrence and neuropathic complications. The increased risk of recurrence observed with mesh fixation possibly results from selection bias. Large randomized controlled studies are needed to determine whether mesh fixation is truly related to neuropathic complications and the incidence of recurrence. PMID- 11727080 TI - Minimally invasive video-assisted approach for partial and total thyroidectomy: initial experience. AB - BACKGROUND: We report our initial experience with partial and total thyroidectomy using a video-assisted approach. The feasibility, safety, and potential benefits of this technique are examined. METHODS: Between January and May 2000, 28 patients were select to undergo a thyroid lobectomy (n = 17) or total thyroidectomy (n = 11) by a video-assisted cervical approach. Patient selection was based on clinical examination and preoperative ultrasonography. The surgical procedures were conducted under general anesthesia through a minimal substernal skin incision. Frozen sections were examined peroperatively in all cases. RESULTS: The initial diagnosis was solitary nodule in 19 patients and multinodular goiter in 8 patients. One patient was treated for hyperthyroidism. The mean cranio-caudal axis and transverse diameter of the resected specimen were 4.9 +/- 0.9 and 2.7 +/- 0.5 cm, respectively, and the mean total lobar weight was 11.9 +/- 5.5 g. Conversion to conventional surgery was required in three patients (10.7%), due to local bleeding in all cases. The mean operative times were 150 +/ 8.2 and 102.5 +/- 17 min for total and partial thyroidectomy, respectively. The laryngeal nerve was identified in 94.8% of cases. The mean length of skin incision was 25.4 +/- 2 mm. There was one case of postoperative hypocalcemia and one case of postoperative hoarseness. One patient had a transient vocal cord palsy. The postoperative hospital stay was 1 day for 66.7% of patients. The pain intensity evaluation, performed on postoperative day 1 using the visual analogue scale (VAS) method, was 1.9 +/- 1.4. CONCLUSION: Video-assisted thyroidectomy is feasible, safe, and effective in selected cases. Benefits for the patients in terms of postoperative pain, hospital stay, and cosmesis still need to be assessed in a prospective trial comparing standard open and video-assisted approaches. PMID- 11727081 TI - Video-assisted neck exploration for primary and secondary hyperparathyroidism: initial experience. AB - BACKGROUND: Minimally invasive surgery (MIS) for primary hyperparathyroidism includes unilateral neck exploration, access via a totally endoscopic approach, and access via a video-assisted procedure. We report herein our initial experience with the video-assisted neck exploration procedure for primary (PHPT) and secondary hyperparathyroidism (SHPT). METHODS: Between June 1999 and May 2000, 35 patients were selected for PHPT (n = 25) and SHPT (n = 10). In all cases, video-assisted neck exploration was performed under general anesthesia, leading to selective adenoma removal in PHPT or subtotal parathyroid resection in SHPT. Patient selection was based on the preoperative localization studies (ultrasonography and sestamibi scintigraphy). RESULTS: Five of the 25 patients operated on for PHPT (20%) and three of the 10 patients who underwent surgery for SHPT (30 %) were converted to a conventional surgical technique. The mean operative times in PHPT and SHPT were 48.9 +/- 18.7 min and 136.8 +/- 18.7 min, respectively. The recurrent laryngeal nerve was identified in 45% of the patients. The median size and weight of the resected parathyroid glands were 1.8 cm (range, 0.9-2.5) and 1 g (range, 0.5-7), respectively. The length of skin incision was 24 +/- 1.2 mm. All but one patient are currently cured, with a median follow-up of 6 months (range, 2-13). Postoperative complications included hematoma and transient hoarseness in one patient each (2.85%). The median pain intensity, based on the visual analogue scale (VAS) method, as measured at postoperative day 1 was 0.5 (range, 0-3.6). In the PHPT group, the postoperative hospital stay was PMID- 11727082 TI - Video-assisted parathyroidectomy via the lateral approach vs conventional surgery in the treatment of sporadic primary hyperparathyroidism: results of a case control study. AB - BACKGROUND: We previously demonstrated that minimally invasive video-assisted parathyroidectomy (VAP) can be performed via a lateral approach on the line of the sternocleidomastoid muscle. The aim of this study was to compare the results of this technique with those of conventional parathyroidectomy (CP) in a case control study. METHODS: Over a 2-year period, 80 VAP were attempted. The selection criteria were as follows: sporadic primary hyperparathyroidism, no history of previous neck surgery, no thyroid disease, suggestion of a single adenoma on preoperative imaging. A rapid intraoperative parathyroid hormone (PTH) assay was performed. The procedure was completed successfully in 68 patients. A case-control study of 68 patients who underwent CP for a single adenoma was performed. The controls were matched for age and sex. RESULTS: All of the patients were normocalcemic at follow-up. No statistically significant differences between the VAP and the control groups were found for age, sex, pre- and postoperative calcemia and PTH, adenoma weight, operating time, complication rate, or postoperative stay. One VAP patient developed recurrent laryngeal nerve palsy. Patients who underwent VAP required less analgesics (p < 0.0001) and were more satisfied with the cosmetic results (p < 0.0001). CONCLUSIONS: This study suggests that VAP by the lateral approach has some advantages over CP in terms of postoperative pain and cosmetic results. PMID- 11727083 TI - Video-assisted parathyroidectomy for primary hyperparathyroidism: a new approach involving a skin-lifting method. AB - BACKGROUND: Recent advances have allowed the performance of parathyroidectomy as an endoscopic procedure. Carbon dioxide (CO2) insufflation can be used to create a working space in the anterior neck, but it has been associated with a number of complications. We have devised a skin-lifting method to overcome these problems. METHODS: Eleven consecutive patients underwent video-assisted parathyroidectomy. Preoperative imaging revealed a solitary adenoma in all 11 cases. A 3-cm oblique incision was made below the clavicle, and a 5-mm incision was made on the lateral neck. After the skin was lifted, video-assisted parathyroidectomy was performed. RESULTS: Surgery required 186 +/- 50 min. No conversions to conventional cervicotomy were needed. Levels of serum calcium and intact parathormone decreased significantly in all patients on postoperative day 1. Laryngeal recurrent nerve paresis and seroma were noted in one patient each. CONCLUSIONS: Our procedure eliminates any potential CO2 problems and offers the advantages of direct manipulation and improved cosmesis. Endoscopic parathyroidectomy should be considered a viable option for the surgical treatment of a solitary adenoma. PMID- 11727084 TI - Short-term outcome after laparoscopic and open 360 degrees fundoplication. A prospective randomized trial. AB - BACKGROUND: Despite the lack of randomized trials supporting the laparoscopic approach, laparoscopic antireflux surgery has gained widespread acceptance during the last decade. The aim of this study was to compare the short-term symptomatic and objective outcome after laparoscopic and open 360 degrees fundoplication in a prospective randomized clinical trial. METHODS: Sixty patients with GERD were randomized to undergo either laparoscopic (LF) or open 360 degrees fundoplication (OF). Endoscopy, esophageal manometry, 24-h pH monitoring, clinical symptom evaluation, and symptom scoring according to a validated questionnaire (the Gastrointestinal Symptom Rating Scale [GSRS]) was performed preoperatively and 6 months after surgery. RESULTS: Five patients randomized to the laparoscopic group were converted to open surgery. Esophageal acid exposure was restored to normal in all patients. Lower esophageal sphincter length and resting pressure were significantly increased after both laparoscopic and open fundoplication (p < 0.001); there were no differences between the groups. No significant differences were seen in symptomatic outcome, although there was a trend toward a higher rate of mild dysphagia (p = 0.051) after laparoscopic surgery. GSRS revealed a decrease in reflux score (p < 0.001) and abdominal pain score (p < 0.001) postoperatively. There were no significant differences in GSRS scores between the two groups. CONCLUSION: Laparoscopic 360 degrees fundoplication is as effective in treating reflux disease as open fundoplication. Six months postoperatively, no significant differences were seen in symptomatic or objective outcome. Long-term evaluation is needed. PMID- 11727085 TI - Staging of pancreatic and ampullary cancers for resectability using laparoscopy with laparoscopic ultrasound. AB - BACKGROUND: Cancers of the pancreas and periampullary region are rarely curable. We set out to determine the efficacy of laparoscopy with laparoscopic ultrasound in the staging of pancreatic and ampullary malignancies for resectability. METHODS: Between January 1994 and September 1999, we retrospectively reviewed the laparoscopic staging (LS) of tumors already deemed resectable by standard radiologic criteria in 27 patients using laparoscopy with laparoscopic ultrasound (LUS). Patients found to be resectable by LS evaluation underwent laparotomy (LA). We then compared the results of the LS and LA findings. RESULTS: Of the 27 patients evaluated, 17 were men and 10 were women. Their mean age was 66 years. Preoperative computerized tomography (CT) scans were done in all 27 patients (100%), and transabdominal and endoscopic ultrasound (EUS) was done in 21 (78%). By LS, seven patients (26%) were found to have unresectable disease. Two patients with mesenteric tumor infiltration (one with peritoneal implants, and one with a visible liver metastasis) were judged to be unresectable by laparoscopy alone. LUS revealed that one patient had portal vein (PV) occlusion and two had metastases to the lymph nodes or liver that were not revealed by preoperative studies or laparoscopy alone. Among 20 patients (74%) deemed resectable by LS, two (10%) were found to be unresectable at LA, one due to PV involvement and the other due to local tumor extension with superior mesenteric lymph node metastasis. Eighteen of those in whom resection was attempted (90%) were resectable, with no unexpected findings of distant lymph node or hepatic metastasis. Pathology examination showed that eight had regional metastases (44%). The sensitivity of LS in determining unresectability was 77% (seven true positives and two false negatives). The negative predictive value (reflecting resectability) was 90%. Laparoscopy alone had a sensitivity of 44%, with a negative predictive value of 78%. The sensitivity and positive predictive value of LS was 100%, reflecting no false positive examinations. CONCLUSIONS: LS can effectively stage most patients and reliably predict which of them will benefit from LA. Intervention for unresectable patients can then be limited to laparoscopic or endoscopic bypass. The main limitation is that LS may underestimate PV and regional lymph node involvement. PMID- 11727086 TI - Periampullary tumors. Analysis of 319 consecutive cases submitted to preoperative endoscopic biliary drainage. AB - BACKGROUND: During the last 2 decades, endoscopic retrograde cholangiopancreatography (ERCP) has been widely used for the diagnosis of periampullary tumors and the preoperative or definitive treatment of jaundice. METHODS: We performed a retrospective analysis of 319 consecutive patients (184 men and 135 women with an average age of 66.5 years) who underwent ERCP for periampullary tumors between 1987 and 1999. RESULTS: Endoscopic internal biliary drainage was successful in 293 patients (92%), with some differences due to the origin of the tumor. There were five complications (1.5%), including four bleeds and one retroduodenal perforation. There were no deaths related to the endoscopic drainage. Eighty-four patients underwent pancreaticoduodenectomy. The postoperative morbidity rate was 23%, and the overall mortality rate was 4.8%. CONCLUSION: ERCP is a valid technique for the detailed preoperative assessment of periampullary tumors. It is also a safe method for internal biliary drainage. PMID- 11727087 TI - Dedicated minimally invasive surgery suites increase operating room efficiency. AB - BACKGROUND: The rapid adoption of laparoscopic surgery since the late 1980s added tremendous complexity into the operating room (OR) environment. For each case, a plethora of additional equipment-including monitors, video equipment, wiring, tubing, and cords-had to be set up, prolonging OR turnover time and decreasing OR efficiency. In 1993, the concept of designated minimally invasive surgery (MIS) suites was introduced. MIS suites integrated monitors and video equipment into the OR on ceiling-mounted columns and moved the controls to a centralized nursing station. The overall effect of this innovation on OR efficiency has not been measured. METHODS: Five RNs with varying degrees of MIS experience were instructed on video setup and put-away criteria and then timed while performing a set of standardized tasks. Each set of tasks was performed twice using a standardized surgery model. Differences in setup and put-away times between MIS suites and standard ORs were tested using the t-test for paired comparisons. RESULTS: The mean +/- standard deviation (SD) video setup times were 27.9 +/- 5.3 sec (MIS) and 254.3 +/- 54.0 sec (standard); the put-away times were 19.8 +/- 2.7 sec (MIS) and 222.3 +/- 26.0 sec (standard). The mean difference +/- standard error (SE) in both the setup (226.4 +/- 16.9 sec, p = 0.0001) and put-away times (202.5 +/- 8.6, p = 0.0001) were large and statistically significant. CONCLUSION: Using a simulation model, we have demonstrated that the use of a MIS suite reduces video setup and put-away time significantly, with the potential for significant associated cost savings. This provides just one justification for the high cost of building such "ORs of the future." PMID- 11727088 TI - Laparoscopic treatment of hemorrhage after vaginal hysterectomy or laparoscopically assisted vaginal hysterectomy (LAVH). AB - BACKGROUND: This study examines the use of laparoscopy for the treatment of secondary hemorrhage following vaginal or laparoscopically assisted vaginal hysterectomies (LAVH). METHODS: Over a 5-year period, the incidence and management of postoperative bleeding following vaginal hysterectomies or LAVH were registered prospectively. RESULTS: The overall incidence of hemorrhage after vaginal hysterectomies or LAVH was 1.2% (17 of 1319). Over the 5-year period, it decreased from 2.4% (five of 209) to 0.6% (two of 315). Surgical revision was initiated transvaginally in nine patients and by laparoscopy in eight patients. Five of the eight patients profited from the prompt use of laparoscopy; inconclusive vaginal exploration was followed by laparoscopy in another five patients. CONCLUSION: Hemorrhage following vaginal hysterectomy or LAVH can be treated by laparoscopy in the majority of patients. Laparoscopy is recommended if the source of bleeding cannot be identified clearly by vaginal examination and/or if an intraabdominal bleeding source is suspected. PMID- 11727089 TI - The surgical trauma of abdominal wall incision. A comparison of laparoscopic vs open surgery with three-dimensional stereography. AB - BACKGROUND: Laparoscopic operations seem to respect the integrity of the abdominal wall better than conventional laparotomy, but the effects of surgical trauma are not well understood. The new technique of three-dimensional stereography makes it possible to describe and calculate the mobility of the abdominal wall and the nature of the underlying disturbances. METHODS: Three dimensional stereography is a noninvasive optical method of measuring surface areas. Abdominal wall mobility can be assessed by comparing changes to the abdominal surface in its minimum and maximum excursions. Different parameters, such as height difference and curvature, are calculated. We studied patients undergoing different types of surgical procedures (laparoscopy and open surgery) by measuring their abdominal wall mobility before and after the procedure. We also compared these patients to a control group. Each group consisted of 30 patients, who were evaluated prospectively. RESULTS: We found a significant difference in abdominal wall mobility between patients treated via a laparoscopic approach, and those who had conventional surgery. At 7 days after laparoscopy, abdominal movement was always the same in the laparoscopic group as in the controls. By contrast, the open surgery group still showed a significant lack of mobility 12 days after the procedure. CONCLUSION: The new method of three dimensional stereography makes it possible to compare the trauma associated with different surgical approaches as it affects the integrity of the abdominal wall. As compared with open surgery, laparoscopy has a significant positive effect on abdominal wall integrity. PMID- 11727090 TI - The impact of laparoscopic biopsy of pancreatic lymph nodes on lipid peroxidation using helium and carbon dioxide in BOP-induced pancreatic cancer in hamsters. AB - BACKGROUND: Increased lipid peroxidation is believed to stimulate liver metastasis in pancreatic cancer. However, the effect of the laparoscopic biopsy of pancreatic lymph nodes on liver metastasis is still unknown. We hypothesized that the effects of a pneumoperitoneum with carbon dioxide (CO2) increase lipid peroxidation and stimulate liver metastasis. METHODS: Ductal pancreatic adenocarcinoma was induced in Syrian hamsters (n = 30) by weekly subcutaneous injections of N-nitrosobis-2-oxopropylamine (BOP) for 12 weeks. In group 1, a laparotomy and biopsy of pancreatic lymph nodes was performed. Groups 2 and 3 underwent laparoscopic biopsy with either CO2 or helium. In the 5th postoperative week, the hamsters were killed and the incidence of liver metastasis was analyzed histopathologically. We then made determinations of the level of lipid peroxidation (thiobarbituric acid-reactive substances [TBARS])as well as the activity of glutathionperoxidase (GSH-PX) and superoxidismutase (SOD) in pancreatic carcinoma and liver metastases of the animals. RESULTS: The incidence of liver metastasis was higher in the CO2 group (60%) than in the helium (30%) and laparotomy groups (20%) (p < 0.05). The concentration of TBARS was greater in pancreatic carcinoma and intrametastatic hepatic tissue than in extratumorous pancreatic and extrametastastic hepatic tissue in all groups. Extrametastatic hepatic concentration of TBARS was higher in the CO2 group (19.4 +/- 0.88 nmol/mg protein) than the laparotomy (10.66 +/- 0.95 nmol/mg protein) and helium groups (10.79 +/- 0.58 nmol/mg protein). GSH-PX and SOD activity was significantly lower in pancreatic carcinoma tissue and intrametastatic hepatic tissue than in extratumorous pancreatic and extrametastatic hepatic tissue. However, in the CO2 group (1.24 +/- 0.48 107 U/mg protein), extrametastatic GSH-PX activity was lower than in the laparotomy (1.73 +/- 0.4 107 U/mg protein) and helium groups (1.63 +/ 0.28 107 U/mg protein). CONCLUSION: After laparoscopic biopsy of pancreatic lymph nodes in the CO2 group, lipid peroxidation was increased and GSH-PX activity was decreased in extrametastatic hepatic tissue compared to the intrametastatic sections. This mechanism may be responsible for the increased liver metastasis in the CO2 group. PMID- 11727091 TI - Mobilization of the phrenic nerve in the thoracic cavity by video-assisted thoracic surgery. Techniques and initial experience. AB - BACKGROUND: The use of video-assisted thoracic surgery (VATS) techniques to mobilize the phrenic nerve in the thoracic cavity for neurotization after brachial plexus injury was studied. METHODS: From August 1999 to January 2000, 10 men and 1 woman with brachial plexus injury (left side in 5 and right side in 6) joined the study group. Their ages ranged from 20 to 38 years (average, 28 years). Supine after general anesthesia, all the patients had double-lumen trachea cannulas to guarantee complete lung collapse on the operative side. Three port incisions were made to allow introduction of the following: a 10-mm Stryker endoscope through the sixth intercostal space 2 cm medial to the anterior axillary line, one instrument for manipulation in the anterior axillary line of the third intercostal space, and another in the second intercostal space about 2 cm lateral to the parasternal line. The nerve was mobilized with two common long Mixter clamps and some endoscopic instruments by blunt and sharp dissection. RESULTS: All patients were managed successfully without severe complications. The mean additional length of phrenic nerves by this technique was 16 cm. CONCLUSIONS: Mobilization of the phrenic nerve by VATS is a safe and minimally invasive method for elongating the nerve for neurotization after brachial plexus injury. PMID- 11727092 TI - Prevention of compensatory hyperhidrosis after thoracoscopic sympathectomy for hyperhidrosis. AB - BACKGROUND: Compensatory hyperhidrosis is a troublesome complication of thoracoscopic sympathectomy for hyperhidrosis. After extensive resection of the second through the fourth ganglion (T2-4), as well as after limited resection of the second ganglion (T2), the reported incidence of compensatory hyperhidrosis ranges as high as 50-97%. The purpose of this study was to determine whether the incidence of compensatory hyperhidrosis can be reduced by limiting the thoracoscopic sympathectomy to another level, the third ganglion. METHODS: We analyzed 28 thoracoscopic sympathectomies for palmar and/or axillary hyperhidrosis. In all patients, the sympathetic chain was transected cranially and caudally to the third ganglion (T3 dissection). Long-term follow-up was conducted by interviewing patients using standardized questionnaires. RESULTS: The surgery was effective in all patients. After a median follow-up of 3.5 years, compensatory hyperhidrosis was not recorded in any of the patients. There were no recurrences of hyperhidrosis. CONCLUSION: Limited thoracoscopic sympathectomy at the level of the third ganglion is effective and seems to prevent compensatory hyperhidrosis. PMID- 11727093 TI - Thoracoscopic treatment for delayed presentation of congenital diaphragmatic hernia in the infant. A report of three cases. AB - BACKGROUND: Delayed presentations of congenital hernia occurring outside of the neonatal period have been reported for all ages. Classically, repair of the hernia defect involves a transverse subcostal laparotomy in the pediatric age group and usually a thoracotomy in the adult. The first report describing a laparoscopic repair of a congenital diaphragmatic hernia in a 6-month-old infant was published in 1995. During the past 25 years, 17 patients with delayed presentation of congenital diaphragmatic hernias have been managed by our pediatric surgery team. The last three patients underwent surgery thoracoscopically. PATIENTS AND METHODS: This study involved two boys (ages, 8.3 and 19 months, respectively) and one girl (age, 9 months) weighing 8 to 13 kg. All three infants underwent surgery using a thoracoscopic approach with general anesthesia. A thoracic epidural catheter was placed systematically for postoperative analgesia during the first 24 hours. Reduction of the hernia contents using one optical trocar and two operating trocars was difficult in the case without any hernia sac. In the cases with a hernia sac, reduction was easily and quickly obtained with a pleural insufflation of carbon dioxide (~8 mmHg). The hernia defect was repaired using interrupted sutures of 2/0 Ethibond. For two of the three patients, this repair was reinforced with staples in the one case and a nonresorbable mesh in the other case. RESULTS: The mean operative time was 78 min. The chest tube was removed on the first or second postoperative day and the patients were discharged on the fourth or fifth postoperative day. At the 19 month follow-up assessment in one case, the chest x-ray was perfectly normal and diaphragmatic motion also was confirmed to be normal by ultrasonography. CONCLUSIONS: The thoracoscopic approach for the repair of delayed-presentation congenital diaphragmatic hernia is feasible. Our results demonstrate the safety and efficiency of this surgery, as well as a remarkable functional and cosmetic result and a very quick recovery. PMID- 11727094 TI - An analysis of video-assisted thoracoscopic resection for mediastinal masses in 150 cases. An overview of the pansternal approach, histology, and complications. AB - BACKGROUND: To clarify the usefulness of the diagnosis and treatment of mediastinal masses, by video-assisted thoracoscopic surgery (VATS), we performed a retrospective multi-institutional study to delineate the type of approach, histopathology, and complications associated with these entities. METHODS: We analyzed 150 patients who underwent the VATS procedure at several institutions between 1991 and 1999. RESULTS: VATS resections were performed using various combinations of the "pansternal approach." The unilateral thorax approach was applied in 135 patients, the bilateral thorax approach combined with the suprasternum approach in seven patients, the unilateral thoracic approach combined with the supraclavicular approach in three patients, the infrasternal approach in three patients, and the bilateral thoracic approach combined with both the infrasternal and the suprasternal approaches in two patients. Thoracoscopic resections or biopsies diagnosed 140 benign and 10 malignant mediastinal masses. There were operative complications in nine patients (6%). Eight patients (5.3%) were converted to conventional thoracotomy. CONCLUSION: VATS is a safe, effective, minimally invasive technique that can facilitate the resection of mediastinal masses when the pansternal approach is applied as appropriate for the location of the tumor. PMID- 11727095 TI - Clinical efficacy of the Toupet technique performed by laparoscopic surgery. Medium-term follow-up in 122 cases. AB - BACKGROUND: We present the results obtained in a series of patients with gastroesophageal reflux disease (GERD) who underwent the same type of laparoscopic surgery (Toupet technique). METHODS: A total of 122 patients with symptomatic hiatal hernia and GERD were submitted to laparoscopic surgery between March 1993 and March 1999. In all patients, we performed a preoperative gastroduodenal radiological study, an upper gastrointestinal (GI) endoscopy, esophageal manometry, and 24-h pH monitoring. The same studies were repeated at 3 months. All patients were followed up clinically for ?1 year. The only laparoscopic technique used was the Toupet (270 degrees posterior fundoplication). RESULTS: In 117 patients (95.9%), the clinical results were good, with complete disappearance of the symptoms. Two patients suffered a relapse, which was resolved by reintervention with laparoscopy. One patient experienced dysphagia and frequent chest pain for 5 months. Another two patients had moderate difficulty in belching. The endoscopic and radiological barium studies at 3 months were normal in all patients. Significant differences were seen in functional studies performed prior to and 3 months after the operation in all patients (p < 0.05). CONCLUSIONS: Surgical laparoscopic treatment with the Toupet technique is effective both clinically and functionally. In cases of relapse, a new repair is possible with laparoscopy. Long-term follow-up of these patients is necessary to show whether these results are maintained over time. PMID- 11727096 TI - Video-assisted endoscopic lumbar discectomy. AB - BACKGROUND: The optimal management of symptomatic lumbar disc herniations (LDH) remains controversial. This study examines the feasibility and safety of a video assisted endoscopic intracanalicular technique for managing symptomatic LDH. METHODS: From September 1999 to June 2000, we used the current technique, the Vertebroscope System, on 11 patients (six men, five women), aged from 18 to 61 years (mean, 45), who had suffered symptomatic LDH. The disc levels involved were at L4-L5 (n = 8), and L5-S1 (n = 3). The Vertebroscope, which has a 30 degrees viewing angle and a working channel 1.7 cm in diameter, was used for the minimally invasive endoscopic procedures. The mean follow-up period was 12 months (range, 6-15). RESULTS: The operating time ranged from 60 to 335 min (mean, 136.5), and the estimated blood loss during operation was minimal to 200 ml. The mean length of the paramedian skin incisions was 2 cm. No drainage tube was used postoperatively. The mean hospital stay was 3 days (range, 2-5), with five patients discharged on the 1st postoperative day. Complications included one superficial wound infection, one conversion to an open procedure when muscle herniation into the working channel created a technical difficulty in approaching the ligamatum flavum, and one minor tear of the nerve root sleeve that did not require further surgery. In the first five patients studied herein, the mean operating time was significantly longer than that for the later five patients (201 vs 72 min, p < 0.001). CONCLUSIONS: The advantages of the current endoscopic disectomy technique include its minimally invasive character, with less paraspinal muscle trauma, direct address to the lesion site that resembles the open technique, and enhanced operative field visualization with a paramedian skin incision of just 2 cm. Practice is needed to perfect such an endoscopic approach for lumbar disc excision, so the operating time decreased significantly as the surgeons became more familiar with this endoscopic technique. It has proved to be safe and effective for treating patients with symptomatic LDH. PMID- 11727097 TI - Laparoscopic transperitoneal hernia repair of incarcerated hernias: Is it feasible? Results of a prospective study. AB - BACKGROUND: Laparoscopic transperitoneal hernia repair (TAPP) has proved its efficiency in elective surgery. However, TAPP results with incarcerated hernias still are unknown. METHODS: Data from a prospective study were evaluated with regard to TAPP repair for both chronically and acutely incarcerated hernias. RESULTS: During a 6-year period, 220 incarcerated hernias were repaired (194 via TAPP). The median operation time for TAPP was 55 min. An accompanying resection therapy became necessary for only four of the emergency cases (11.1%) and two of the chronically incarcerated cases (1.3%) in the TAPP group. Postoperative morbidity was 2.8% in the emergency group and 3.8% in the chronically incarcerated group, which does not differ from the rate for TAPP used on reducible hernias. One recurrence was found 26 months after TAPP reconstruction (0.5%). CONCLUSION: Laparoscopic inguinal hernia repair (TAPP) represents an efficient therapeutic concept in the treatment of both chronically and acutely incarcerated inguinal hernias. PMID- 11727098 TI - 5-aminolevulinic acid-induced (ALA) fluorescence for the laparoscopic diagnosis of peritoneal metastasis. AST An experimental study. AB - BACKGROUND: We performed a randomized experimental study in a rat model to evaluate the use of 5-aminolevulinic acid-induced (ALA) fluorescence in the laparoscopic diagnosis of peritoneal metastases of ovarian cancer. METHODS: We injected 103 ovarian adenocarcinoma cells in the peritoneum of 31 rats. One week later, 5-aminolevulinic acid was injected in the peritoneum (100 mg/kg). After 3 h, we examined the rats using a 10-mm endoscope with a mono CCD camera and a light source developed for photodynamic diagnosis. Metastases on the parietal peritoneum were evaluated independently by two surgeons randomly assigned to use either a conventional light mode or the fluorescence mode. RESULTS: The mean number of metastases detected was 2.84 with conventional laparoscopic light and 5.74 with ALA-induced fluorescence (p < 0.0008). Metastases diagnosed by fluorescence were confirmed by pathologic examination. Random peritoneal biopsy specimens taken from nonfluorescent areas were negative. CONCLUSION: In this experiment, ALA-induced fluorescence improved the detection of peritoneal metastases of ovarian cancer. PMID- 11727099 TI - A 10-year experience with the use of laparoscopic cholecystectomy for acute cholecystitis: is it safe? AB - BACKGROUND: In the era of open surgery, emergency open cholecystectomy has been shown for many reasons to be preferred to delayed surgery for acute cholecystitis. Despite the fact that elective laparoscopic cholecystectomy (LC) has become the gold standard for the treatment of symptomatic gallstone disease, the same procedure remains controversial for the management of acute cholecystitis because it is considered to be associated with more complications and an increased risk of common bile duct injuries than interval LC after resolution of the acute episode. The purpose of this report is to describe our experience with LC for acute cholecystitis during a 10-year period. METHODS: Patients undergoing laparoscopic surgery have been entered prospectively into a database since 1995. Those who underwent surgery before 1995 were added retrospectively to the same database. Patients were included in this study if they underwent emergency laparoscopic cholecystectomy for suspected acute cholecystitis. The diagnosis was based on clinical, laboratory, and echographic examinations. Analysis was performed to identify risk factors associated with conversion or morbidity. RESULTS: Of the 1,212 patients subjected to LC between 1990 and 1999, 268 (151 women and 117 men), with a mean age of 53 years, underwent surgery on an emergency basis for suspected acute cholecystitis. Their mean age (p = 0.002) and the proportion of men (p < 0.001) were higher than in the elective group. Delay before admission and surgery varied widely, but 72% of the patients underwent surgery within 48 h of admission. An intraoperative cholangiography, attempted in 218 patients, was successful in 207 (95%). Histologic examination confirmed acute cholecystitis in 82% of the patients. Conversion was necessary in 15.6% of the cases. It occurred more frequently in patients who underwent surgery later than 48 (p = 0.03) or 96 h (p = 0.006) after admission. No other predictor of conversion was found. Overall morbidity was 15.3%, and major morbidity was 4.4%. The only risk factor for morbidity was a bilirubin level greater than 20 mmol/l (p = 0.02). Three partial lesions of the common bile duct occurred. All were recognised and repaired immediately with no adverse effect. There was no difference in the overall rate of biliary complications between the patients operated for acute cholecystitis and those who underwent elective surgery. No reoperation was necessary, and there was no mortality. CONCLUSIONS: Although LC is safe and effective for acute cholecystitis, its associated morbidity and conversion rate are higher than for elective LC. The conversion rate decreases with experience. When surgery is performed within 2 or maximally 4 days of admission, in experienced hands, LC represents the treatment of choice for acute cholecystitis. Intraoperative cholangiography should be performed in every case because it helps to clarify the anatomy and allows for early diagnosis and repair of bile duct injuries. PMID- 11727100 TI - Endoscopic marking: an adjunct to laparoscopic gastrointestinal surgery. AB - BACKGROUND: In 1975 one of us (J.P.) first described endoscopic marking of the intestinal lumen with india ink to produce a serosal tattoo, thus facilitating the location of lesions during subsequent surgery. The use of the technique in concert with laparoscopic surgery is particularly important because the ability to palpate the bowel is lost. Carbon-based products such as india ink provide permanent marking. An alternative marker is methylene blue, whose mark is only temporary but clearly seen laparoscopically. METHODS: We have used methylene blue for intraoperative marking, reserving India ink for instances in which marking endoscopy occurs before surgery. RESULTS: 15 patients, over a period of 2 years, had endoscopic marking performed preoperatively or intraoperatively. All markings were easily visualized upon laproscopy, assisting identification of the sites of the lesions. Numerous other cases were marked at endoscopy but did not come to surgery. CONCLUSION: Endoscopic marking is an invaluable tool in the performance of laparoscopic gastrointestinal surgery. PMID- 11727101 TI - Performing laparoscopic surgery is significantly more stressful for the surgeon than open surgery. AB - BACKGROUND: The awkward visual and physical interface of video-endoscopic surgery (VES) has been shown to increase the physical workload of the surgeon, yet there is a lack of objective information on the mental effort and stress demanded by VES techniques. This study addresses the hypothesis that VES is more stressful than open surgery using an ergonomic analysis of surgical tasks in a laboratory setting. METHODS: A portable ergonomic work station was developed using a software Virtual Instrument (VI) interfaced with electronic hardware to compare the mental workload of 28 surgeons. The task was knot tying. The independent variable was work condition: rest, open technique, or VES technique. Dependent variables were tonic skin conductance level (SCL), electrooculogram (EOG), and subjective reports of concentration (CON) and stress (STR). Statistical analysis used nonparametric methods. RESULTS: Subjects tied fewer knots using the VES technique (p < 0.05). The SCL increased progressively from rest to the open task to the VES task (p < 0.05), correlating with the subjects' reported increase in mental stress level (p < 0.05). Eye blinks decreased from rest to the open task (p < 0.05), consistent with the subjects' reported increase in level of mental concentration. From the open to the VES task, eye blinks increased (p < 0.05), as would be expected given the greater demands of the VES task. Experienced subjects demonstrated less variability in SCL levels across tasks. CONCLUSIONS: VES technique requires greater concentration and places greater mental stress on surgeons than to open surgery. More experience with VES may decrease this effect. Studies are needed to improve the human-technology interface the stress. PMID- 11727102 TI - Needlescopic surgery. A logical evolution from conventional laparoscopic surgery. AB - BACKGROUND: The purpose of this study was to analyze the safety and feasibility of needlescopic surgery and to compare the short-term outcomes relative to conventional laparoscopic surgery. METHODS: Needlescopic surgery patients were compared to matched cohorts of conventional laparoscopic surgery patients from the same prospective database for a variety of selected procedures. RESULTS: A total of 101 needlescopic procedures were analyzed (30 cholecystectomy, 28 Nissen fundoplication, 12 bilateral sympathectomy, 10 splenectomy, 10 Heller myotomy, three adrenalectomy, two colon resection, two splenic cyst excision, four other). There was no significant difference between the needlescopic and conventional laparoscopic groups in conversion rates, morbidity, or mortality. A higher proportion of patients were in hospital PMID- 11727103 TI - Selective use of endoscopic retrograde cholangiopancreatography to facilitate laparoscopic cholecystectomy without cholangiography. A review of 1139 consecutive cases. AB - BACKGROUND: The aim of this study was to show that laparoscopic cholecystectomy can be performed safely without routine intraoperative cholangiography. METHODS: We performed a retrospective analysis of 1139 consecutive patients (376 men and 763 women with an average age of 51.4 years) who underwent laparoscopic cholecystectomy between 1991 and 1999. In all, 227 patients (20%) were selected to undergo preoperative endoscopic retrograde cholangiopancreatography (ERCP) on the basis of four criteria for risk of stones. RESULTS: ERCP allowed us to make a diagnosis of biliary stones in 53.3% of the selected patients. Extraction of the stones was successful in 97% of the cases. In 14% of cases, ERCP was normal; in 32.7%, some useful diagnostic information was obtained. There were three complications (pancreatitis) following endoscopy (complication rate, 1.3%). Laparoscopic cholecystectomy was successful in 92% of patients. The postoperative morbidity rate was 3.2% (major complications, 0.5%). There were no deaths. During a follow-up period ranging from 3 to 97 months, six patients (0.6%) were found to have residual biliary stones. CONCLUSION: This study confirms the hypothesis that laparoscopic cholecystectomy can be performed safely without routine intraoperative cholangiography. PMID- 11727104 TI - Laparoscopic management and clinical outcome of emphysematous cholecystitis. AB - BACKGROUND: As opposed to acute, chronic, and acalculus cholecystitis, emphysematous cholecystitis (EC) is associated with significant morbidity and mortality. Only a few studies have specifically reviewed the operative management and clinical outcome of EC. This study documents the operative management and clinical outcome of EC at the Cleveland Clinic Foundation. METHODS: Between January 1996 and June 1999, 18 consecutive patients underwent cholecystectomy for emphysematous cholecystitis at our institution. All charts were reviewed retrospectively, and patients undergoing concurrent procedures were excluded. Mean values +/- standard deviation (SD) of the mean were calculated for patient demographics, preoperative white blood cell count (WBC), bilirubin, alkaline phosphatase, and length of hospital stay. Operative procedure (laparoscopic, converted, or open), preoperative imaging studies, operative time, ICU stay, morbidity, and mortality were reviewed. RESULTS: Patients presented with a mean age of 53.4 years (range, 18-80) and a male/female ratio of 3.5 (14/4). There were no differences between groups in terms of patient demographics. Mean WBC on admission was 14.2 K/mL (range, 5.4-19.7). Mean alkaline phosphatase and total bilirubin were 115 U/L (range, 45-428) and 1.4 mg/dl (range, 0.5-3.4), respectively. Thirteen patients (72%) were completed laparoscopically, two patients (11%) were converted to an open procedure, and three patients (17%) had open surgery. Overall mean length of hospital stay was 5 days (range, 1-18). Two patients from the open group ultimately died 1 year later due to progression of preexisting illness. One of these patients had congestive heart failure and chronic renal failure; the other had metastatic malignant melanoma. None of the patients died in the immediate perioperative period. There were five complications (27.8%). Two patients presented with bleeding secondary to heparin and coumadin therapy. One developed sepsis, and another developed leakage from the cystic duct stump, necessitating an endoscopic retrograde cholangiopancreatogram (ERCP) with stent decompression. The fifth complication, ileus, required readmission 3 days postoperatively, but the patient responded to conservative management. Complications were evenly distributed between the three groups. There were two complications in the laparoscopic group, two in the open group, and one in the conversion group. All other patients were alive at the time of this publication. CONCLUSIONS: Using current techniques, patients with EC can be managed successfully utilizing laparoscopy. Morbidity and mortality appear to be slightly higher than published reports for acute, chronic, and acalculus cholecystitis. Conversion rates are comparable to patients with acute and chronic cholecystitis who undergo laparoscopic cholecystectomy. Based on these data, laparoscopic cholecystectomy should be considered the first-line treatment for patients with known or suspected EC. PMID- 11727105 TI - A technique for laparoscopic-assisted percutaneous drainage of infected pancreatic necrosis and pancreatic abscess. AB - BACKGROUND: Percutaneous drainage has been shown to be an acceptable method for treating both pancreatic abscesses and infected pancreatic necrosis. However, percutaneous techniques have certain shortcomings, including the time and labor required and failure of the catheters to adequately drain the particulate debris. Growing experience around the world indicates that there is a role for retroperitoneal laparoscopy as a means of facilitating the percutaneous drainage of infected pancreatic fluid collections and avoiding a laparotomy. Our technique is discussed in this paper. METHODS: Once infection is documented in a pancreatic fluid collection by fine-needle aspiration, one or more percutaneous drains are placed into the fluid collection(s). A computed tomography (CT) scan is repeated. If further drainage is indicated, retroperitoneoscopic debridement is performed. Using a combination of the percutaneous drain(s) and the post-drain CT scan, ports are placed and retroperitoneoscopic debridement of the necrosectum is performed under direct visualization. Prior to completion of the operation, a postoperative lavage system is created. RESULTS: Six patients with infected pancreatic necrosis have been treated with this technique. Prior to commencement of our laparoscopic protocol, all six patients would have required open necrosectomy. Four of the six patients were managed with retroperitoneoscopic debridement and catheter drainage alone. Complications included a colocutaneous fistula and a small flank hernia. There were no bleeding complications and no deaths. CONCLUSION: Although open necrosectomy remains the standard of care for the treatment of infected pancreatic necrosis and pancreatic abscess, there is growing evidence that laparoscopic retroperitoneal debridement is feasible. PMID- 11727106 TI - Combined endoscopic and laparoscopic removal of ingested large foreign bodies. Case report and decisional algorithm. AB - Foreign body ingestion is a rare occurrence, but quite more frequent in children; adults with mental disorders, bulimia, and dentition alterations; and prison inmates. Most foreign bodies pass spontaneously through the first digestive tract and can move along the entire alimentary canal without giving rise to any major damage. On the contrary, when foreign bodies, because of their shape, size or potential for danger, must be removed, endoscopy is the method of choice. In a small percentage of cases, however, it may be necessary to remove the foreign body surgically. In such cases, the laparoscopic technique certainly can prove to be a most valuable tool, and can be recommended as a first approach. We describe a case in which a large foreign body was accidentally ingested. After a brief analysis of the data in the literature, we propose the decision-making algorithm that we follow. PMID- 11727107 TI - Laparoscopic diagnosis and treatment of idiopathic segmental infarction of the greater omentum. Case report. AB - Acute pain at the right side of the abdomen rarely is caused by idiopathic segmental infarction of the greater omentum (ISIGO). In most cases the patient is presumed to suffer from appendicitis or cholecystitis. Although some radiologic signs might suggest ISIGO, this rare clinical entity mostly is diagnosed perioperatively and confirmed by postoperative pathologic findings. In the reported case, a patient is described with acute right-side abdominal pain of unknown origin, in whom ISIGO was encountered during diagnostic laparoscopy and successfully resected. Because of this minimally invasive approach, the patient was discharged the day after surgery and returned to work after 5 days. The pathogenesis, symptoms, and treatment methods are discussed. PMID- 11727108 TI - Aortic graft in the jejunum without bleeding. A real surprise at endoscopy. AB - It is known that prosthetic infection, graft-duodenal fistula, and erosion are possible late complications after aortic reconstruction, and that all these reported complications are accompanied generally by variable bleeding with different presentations. We report the case of a 63-year-old man who underwent a diagnostic upper gastrointestinal endoscopy for investigation of nausea, anorexia, asthenia, fever, and mild leukocytosis. The patient's medical history included a gastric resection for ulcer, with Billroth II gastrojejunostomy reconstruction and implantation of a Dacron vascular graft for abdominal aortic aneurysm 20 years and 3 years earlier, respectively. Abdomen ultrasonography showed hypoechoic area around an aortic prosthesis. Endoscopy found a foreign body corresponding to the vascular graft at the jejunum. No signs of bleeding were recorded. The patient was hospitalized and submitted to surgery that involved extra-anatomic axillofemoral bypass, bowel resection with a gastrojejunum Roux anastomosis, and prosthesis removal. PMID- 11727109 TI - Management of narrower anastomotic colonic strictures. Case report and proposal technique. AB - Management of narrower (<5-mm) colonic anastomotic stricture mainly is performed endoscopically by repeated balloon dilation often ineffectively. The use of metal self-expanding stents in the malignant and benign stricture of the large bowel has been suggested only recently, and is still being debated. In this report we propose a single-stage procedure that we developed to manage narrower anastomotic colonic stricture. A 60-year-old man 2 years previously had undergone surgery for perforated sigmoid diverticulitis by means of Hartmann's procedure. He was submitted to two mechanical recanalization attempts, both of which failed with dehiscence of anastomoses. He reached us with a significant stricture of the colorectal anastomoses (smaller than 5 mm in diameter) and a diversion ileostomy. After two endoscopic balloon dilations, we observed the relapse of the anastomotic stricture, so we decided to draw up another strategy. We performed a dilation with a TTS balloon, leaving a metallic self-expanding covered stent in situ for 3 months. The aim was to achieve the definitive healing of the anastomotic scar tissue at the desired diameter. We removed the stent during the ileostomy closure. At the time of this writing, 18 months of follow-up evaluation, the patient defecates without any problem PMID- 11727110 TI - Laparoscopic resection of calcifying fibrous pseudotumor of the gastric wall. A unique case report. AB - Calcifying fibrous pseudotumor (CFPT) is a recently described peculiar lesion characterized by the presence of abundant hyalinized collagen with psammomatous or dystrophic calcifications and a lymphoplasmacytic infiltrate. Initially, a few cases of CFPT were observed in the subcutaneous and deep soft tissues, especially at the level of the extremities, trunk, scrotum, groin, neck, and axilla; more recently, several cases have been reported in the pleura. We present a case of CFPT of the gastric wall. To our knowledge, this is the first such case to be described. A previously healthy 49-year-old woman presented with a 5-month history of epigastric pain after eating, early satiety, and weight loss. Gastroscopy revealed a polypoid lesion covered by normal mucosa along the great curvature of the gastric body. The patient was submitted to surgical treatment consisting of laparoscopic wedge resection of the gastric body. Histologic examination of the specimen disclosed CFPT affecting the wall of the gastrointestinal tract. The postoperative course was uneventful, and the patient was discharged from the hospital 5 days after surgery. The patient was free of disease 6 months after excision. Herein we discuss the clinicopathological features and surgical treatment of this most unusual case. PMID- 11727111 TI - Anaphylactoid reaction to intraoperative cholangiogram. Report of a case, review of literature, and guidelines for prevention. AB - Anaphylactoid (pseudoallergic, idiosyncratic) reactions are a well recognized but uncommon consequence to radiographic contrast media. Most reported reactions are to intravascular injections, but systemic reactions to nonvascular injections of radiographic contrast also are well documented. Reactions to nonvascular radiographic contrast media have been reported during or after instillation of radiographic contrast into a multitude of nonvascular body compartments, but not with intraoperative cholangiogram. We describe a case of a systemic anaphylactoid reaction caused by intraoperative cholangiogram during laparoscopic cholecystectomy. We then discuss the clinical presentation, suspected etiology, and treatment of these idiosyncratic reactions as well as established guidelines for prevention in patients at risk. PMID- 11727112 TI - Laparoscopic management of a gastric diverticulum. AB - Gastric diverticuli are rare entities that may present with a variety of vague abdominal symptoms. Diagnosis requires endoscopic and radiologic evaluation preceded by clinical suspicion. This is the first report describing the laparascopic excision of a gastric diverticulum. A description of this procedure and suggestions for the workup of gastric diverticuli are presented. PMID- 11727113 TI - Video-assisted thoracic lobectomy by the substernal hand-assisted method for a metastatic lung tumor. AB - In lobectomy by video-assisted thoracic surgery (VATS), the surgeon is required to have experience and a high degree of skill in performing the operation. We applied substernal hand-assisted thoracoscopic lobectomy to remove a metastatic lung tumor that originated from colon cancer in one patient and obtained satisfactory results. This method compensates for a current limitation of VATS lobectomy, namely, the fact that the surgeon cannot touch the lesion. The reported method is safe and useful for thoracoscopic surgery. It can be performed easily, even if the surgeon is not experienced in performing VATS lobectomy. PMID- 11727114 TI - Late-onset biliary leakage after laparoscopic cholecystectomy using laparoscopic coagulating shears. Report of a case. AB - We recently encountered a rare case of late-onset biliary leakage after laparoscopic cholecystectomy using laparoscopic coagulating shears (LCS). The patient was a 49-year-old Japanese man who had undergone a laparoscopic cholecystectomy at Hamamatsu Medical Center after a diagnosis of cholecystolithiasis associated with localized adenomyomatosis. The cystic duct and the cystic artery were closed using LCS instead of metal endoclips. The postoperative course was uneventful, and the patient was discharged on the 4th operative day. However, on the 7th day after the surgery, the patient developed severe upper abdominal pain and was readmitted to our center with the diagnosis of a late biliary leakage, which was confirmed by an endoscopic retrograde cholangiogram. We then treated the leak successfully with endoscopic nasobiliary drainage. PMID- 11727115 TI - Hand-assisted thoracoscopic resection for anterior mediastinal masses. AB - We report three cases of anterior mediastinal disease (pericardial cyst, thymoma, and myasthenia gravis) in which we used a hand-assisted method in combination with thoracoscopic resection via an infrasternal approach in order to improve the maneuverability and safety of the operation. This technique combines the advantages of both thoracoscopic and incisional resection. It shows promise of becoming a less invasive operation that can be used to treat certain mediastinal tumors, both malignant and benign. PMID- 11727116 TI - Cosmetic minilaparoscopic cholecystectomy. AB - BACKGROUND: Since its introduction in 1987, the technique of laparoscopic cholecystectomy has continued to undergo evolution. One area of refinement has been the optimization of cosmetic results. Surgeons have reduced port size and number or both in attempts to achieve this goal. In this report, we describe a method of adjusting port position to obtain more discreet scars. METHODS: Minilaparoscopic cholecystectomy is performed using three 5-mm ports. One port is placed in the umbilicus. Instead of placing the two additional ports in the right subcostal and subxiphoid positions, they are moved to either side of midline at the level of the pubic hairline. RESULTS: The result is one scar hidden in the umbilicus, with the two other scars located below the bikini line. These scars are nearly undetectable when the patient is wearing minimal clothing. CONCLUSION: We conclude that, in addition to minimizing port size and number, positioning of ports can be used to optimize cosmetic results. PMID- 11727117 TI - A curved edge moderates high pressure generated by a laparoscopic grasper. AB - BACKGROUND: Very high pressures are generated at the interface between the tip of laparoscopic graspers and tissue. The pressure profile suggests that the high pressure is due to stress concentration at the edge of the jaw. Stress concentration at an edge can be modified by curving the edge. This study sought to determine the effect of a curved edge on pressures generated by the jaw of a laparoscopic grasper. METHODS: Pressure generated at the instrument tissue interface was measured by a thin film pressure transducer. The two instruments that we compared were identical in every respect except the profile of the jaw edge. One was unmodified; the other had a rounded grasping edge. Load and handle pressure were constant throughout. Tissue pressures generated by the grasper were recorded as the angle of load was increased from vertical (0 degrees ) to 135 degrees. RESULTS: Localized pressures at the tip of the unmodified instrument reached a maximum of 1500 kPa. Pressures at the tip of the curved instrument reached a maximum of 920 kPa under identical conditions. CONCLUSION: High pressures generated at the tip of laparoscopic graspers can be reduced by rounding the edge of the jaw. PMID- 11727118 TI - Transballoon trocar insertion during preperitoneal hernia repair. AB - Trocar insertion during laparoscopic preperitoneal hernia repair (TEP) can be troublesome because the space into which the trocars are inserted is smaller than that available for transabdominal approaches. Insertion of the trocars directly into the balloon used to dissect the preperitoneal space can facilitate this process. The insertion of a 5-mm trocar into the balloon does not usually result in balloon deflation, and a second trocar can be placed into the balloon as well. Removing the balloon, despite the trocars inside it, is straightforward, allowing the placement of a cannula at the balloon insertion site and initiation of the hernia repair. PMID- 11727119 TI - Influence of nitrous oxide anesthesia on venous gas embolism with carbon dioxide and helium during pneumoperitoneum. PMID- 11727121 TI - Laparoscopic treatment of severe acute pancreatitis. PMID- 11727123 TI - Laparoscopic treatment of ventral hernia. PMID- 11727124 TI - Warmed insufflation carbon dioxide gas for laparoscopic cholecystectomy. PMID- 11727125 TI - Laparoscopic splenectomy. PMID- 11727127 TI - Pulmonary arterial balloon occlusion during video-assisted thoracoscopic surgery for reduction of the risk of massive hemorrhage. PMID- 11727129 TI - Laparoscopic interval appendectomy for periappendicular abscess. PMID- 11727130 TI - Randomized controlled trial of laparoscopic transabdominal preperitoneal hernioplasty vs Shouldice repair. AB - BACKGROUND: There is a scarcity of data on long-term results after laparoscopic hernia repair. Herein we report on the outcome of a group of patients who were followed up for 5 years in a multicenter study on hernia repair. METHODS: A total of 100 patients with 127 hernias were randomized to undergo either transabdominal preperitoneal (TAPP) or Shouldice hernia repair. Follow-up was by clinical examination and standardized questionnaire. RESULTS: Of the 100 patients who underwent surgery, 84 were available for follow-up at 5 years. The TAPP procedure was less painful than the Shouldice repair, with fewer patients receiving narcotic analgesics. The median time to return to 100% activity was shorter in the laparoscopic group (21 days) than in the Shouldice group (40 days). Up to 60 months after the operation, the complication rate was lower in laparoscopically repaired hernias (19/66) than in the open group (25/61). There were two recurrences (3.9%) in the TAPP group and five in the Shouldice group (10.2%). CONCLUSION: The TAPP hernia repair yields comparable or better results than Shouldice herniorrhaphy in terms of postoperative pain, recovery, and recurrence rate. PMID- 11727131 TI - Gastrointestinal quality of life in patients with symptomatic or asymptomatic cholelithiasis before and after laparoscopic cholecystectomy. AB - BACKGROUND: There is an ongoing need, from both the medical and the economic perspective, for a more accurate definition of the influence of symptomatic or asymptomatic gallstone disease on gastrointestinal symptomatology, as well as on the health of the individual in general. METHODS: Using the Gastrointestinal Quality of Life Index (GIQLI), 37 symptomatic and 30 asymptomatic gallstone patients were evaluated at admission to the hospital and again 4 months after undergoing an uneventful laparoscopic cholecystectomy (LC). RESULTS: Postoperatively, significant increases in the total GIQLI score were noted in both the symptomatic group (113.42 +/- 21.9 vs 80.32 +/- 19.1 preoperatively; p < 0.05) and the asymptomatic group (96.37 +/- 14.26 vs 113.30 +/- 15.22; p < 0.05). For the subgroups of items, the core symptoms and the physical, psychological, and disease-specific items improved significantly in both groups in the postoperative period (p < 0.05 for all comparisons), but only the symptomatic group achieved a significant improvement in the subgroup of social items (p < 0.05). Negative correlations were found in both the symptomatic and asymptomatic groups between the preoperative GIQLI scores and the improvement seen after LC (r = -0.70 and r = -0.49, respectively). CONCLUSION: Gallstone disease has a profoundly negative impact on quality of life, especially in symptomatic patients with a history of biliary colic attacks and/or the complications of the disease. Although the condition is not equally distressing for the asymptomatic group of patients without such a history, uncomplicated LC improves the quality of life significantly in both groups. Gallstone patients with lower GIQLI scores are more likely to benefit from LC. PMID- 11727132 TI - Pancreatic complications following laparoscopic splenectomy. AB - BACKGROUND: Laparoscopic splenectomy (LS) has been widely accepted despite a paucity of outcome data. Therefore, we performed a review of LS to assess the pancreatic complications and outcomes associated with this procedure. METHODS: Ninety-four splenectomies were performed for a variety of hematologic disorders. The patient was placed in the lateral position, and three or four trocars were used. RESULTS: LS was completed successfully in 93 patients. One case was converted to an open splenectomy for suspected gastrotomy. Thirty of 32 patients with splenomegaly underwent successful LS. Fifteen patients (16%) had some evidence of pancreatic injury. Six patients had asymptomatic hyperamylasemia. An injury directly associated with an adverse outcome occurred in nine cases (9.5% overall); six patients had pancreatic collections, one had a pancreatic fistula, and two developed hyperamylasemia and pain altering the length of hospitalization. Four of these nine patients did not have elevated postoperative amylase levels and were readmitted with pancreatic complications. CONCLUSIONS: LS can be performed for most pathologic conditions. Pancreatic injury is the most common morbidity associated with LS. The detection of hyperamylasemia can alert the surgeon to a pancreatic injury and alter postoperative management. PMID- 11727133 TI - Hand-assisted laparoscopic surgery: an emerging technique. AB - BACKGROUND: Some surgeons are finding that the placement of one hand in the abdomen during laparoscopic procedures returns tactile feedback lost during purely laparoscopic surgery and facilitates dissection, retraction, and control of bleeding. Studies comparing patient postoperative discomfort after laparoscopic and hand-assisted laparoscopic procedures have not found a significant difference. METHODS: This article is a review of the current literature on hand-assisted laparoscopic surgery and of the different hand assisted devices on the market. Included in the review are opinions of expert laparoscopic surgeons who have used hand-assisted devices. RESULTS: More than 100 hand-assisted laparoscopic procedures have been described in the literature. At least four different companies are involved in hand-assisted laparoscopic devices. Three of these companies currently are Food and Drug Administration (FDA) approved in the United States. CONCLUSIONS: Hand-assisted laparoscopic surgery is not necessary for all laparoscopic procedures. Hand-assisted laparoscopic technique is advantageous for certain procedures and clinical situations such as en bloc resections and removal of solid organ tumors, large colon tumors, and the kidney after donor nephrectomy. This technique offers benefits when a large incision is necessary to complete surgery such an open colon anastomosis. PMID- 11727134 TI - Influence of three-dimensional vision on surgical telemanipulator performance. AB - BACKGROUND: Different viewing conditions (two- and three-dimensional National Television Standard Committee [2D-NTSC and 3D-NTSC] and two-dimensional high definition television [2D-HDTV]) on telemanipulator performance were evaluated. METHODS: Six taskes were performed by 15 endoscopic surgeons using the daVinci telemanipulation system. Performance time and errors were measured. Encoder data from the system were used for kinematic analysis of motion. A self-evaluation questionnaire regarding performance under various viewing conditions was obtained. RESULTS: Resolution was better with 2D-HDTV. The estimate of relative distance was not influenced by the different visualization systems. Motor skill tasks were performed faster with binocular vision (3D-NTSC) than with monocular vision (2D-NTSC, 2D-HDTV). For both 2D settings, the deceleration phase of motion was prolonged (p < 0.05 vs 3D). Peak velocity was reduced with 2D-HDTV as compared with 3D-NTSC (p = 0.01). The surgeons tended to favor the 3D system despite their use of 2D systems in their own practice. CONCLUSIONS: Three dimensional vision enhances telemanipulator performance as compared with a 2D system at the same or higher level of resolution. Because it allows faster and more precise movement, future surgical systems should focus on 3D visualization. PMID- 11727135 TI - Changes in T cell subsets, interleukin-6 and C-reactive protein after laparoscopic and open colorectal resection for malignancy. AB - BACKGROUND: Attenuation of the immune response to surgery, as demonstrated with the laparoscopic approach to cholecystectomy, has potential benefits in patients undergoing laparoscopic colonic resection for malignancy. We aimed to study the perioperative immune response in patients undergoing laparoscopically assisted and open surgery for colorectal cancer. METHODS: This study involved 23 patients undergoing laparoscopically assisted (n = 13) and open surgery (n = 10). Interleukin-6 (IL-6) C-reactive protein (CRP), the total lymphocyte count, and the CD3, CD4, CD8, CD16, and CD19 lymphocyte subpopulations were assayed preoperatively and at 4, 8, 10, 24, 48, and 168 h postoperatively. RESULTS: Significant rises in IL-6 and CRP were demonstrated within 4 and 24 h, respectively (p < 0.001) in both groups. However, no significant difference between the groups was seen. Significant decreases in total lymphocyte count and all T cell subsets were demonstrated in both groups, beginning at 4 h (p < 0.01). However, no significant difference between the groups was seen. All parameters, excluding CRP, had returned to baseline by 7 days postoperatively in both groups. CONCLUSIONS: Patients with malignancy exhibit significant perioperative immune disturbance with laparoscopically assisted and open surgery. The current data do not provide justification for the laparoscopically assisted approach on grounds of immune preservation. PMID- 11727136 TI - Histologic evaluation of fetal brains following maternal pneumoperitoneum. AB - BACKGROUND: The purpose of this study was to determine if maternal pneumoperitoneum with carbon dioxide (CO2) produces evidence of central nervous system (CNS) injury in preterm fetal guinea pigs. METHODS: Thirty pregnant guinea pigs at gestational day (GD) 45 were assigned at random to one of three treatment groups: anesthesia only, CO2 pneumoperitoneum (5 mmHg), or laparotomy. Dams were killed 3 or 5 days postprocedure and fetal brains (83 total) harvested and fixed for subsequent histopathologic evaluation. For comparative purposes, histologic features of fetal guinea pig brain injury were defined from examination of fetal brains harvested from an additional dam that underwent laparotomy with 20 min of uterine arterial occlusion. RESULTS: Carbon dioxide pneumoperitoneum did not increase maternal/fetal morbidity. No evidence of brain injury was found in fetuses from any of the treatment groups. CONCLUSION: Carbon dioxide pneumoperitoneum at 5 mmHg for 40 min in the pregnant guinea pig does not produce evidence of fetal brain injury. PMID- 11727137 TI - A prospective study of bile leaks after laparoscopic cholecystectomy. AB - Since laparoscopic cholecystectomy rapidly became the gold standard, there is an increased morbidity of 1% to 3% for clinically significant bile leaks with this procedure, as compared with open cholecystectomy (<1%). The identification of subclinical bile leaks using cholescintigraphy occurs in the range from 31.4% to 40% after elective open cholecystectomy. At this writing, no studies exist that document the rate of subclinical bile leaks after elective laparoscopic cholecystectomy. In this study, 71 patients were evaluated using cholescintigraphy after elective laparoscopic cholecystectomy. This study represents the first prospective look at the rate of subclinical bile leaks after laparoscopic cholecystectomy in elective cases, and the findings show an overall incidence of 7.3%, as compared with historical reports of 30% to 44% for open cholecystectomy. PMID- 11727138 TI - Definitive treatment of extended Hirschsprung's disease or total colonic form. AB - BACKGROUND: Between December 1990 and March 1999, five laparoscopic Duhamel pull through procedures for extended or total aganglionosis were performed in our department, one of which had a rectosigmoid form with a short bowel and a colonic resection due to a volvulus. METHODS: The aim of this study was to show that even when the extended form of Hirschprung's disease or anatomic difficulties such as a short bowel and anterior colonic resection are encountered, the laparoscopic approach is possible and total colectomy is feasible. The procedure has been described previously. We used one camera port and three working ports. The sigmoid, transverse, and right colon up to the last ileal cove were mobilized laparoscopically. A standard posterior ileo-anal anastomosis was performed, and an endo-GIA stapler was used for the anterior anastomosis. RESULTS: A total of five patients underwent laparoscopic surgery for Hirschprung's disease. There were three total colonic forms, one rectosigmoid form with a short bowel and colon resection attributed to a volvulus, and one transverse variant that required a Deloyers' maneuver for the pull-through. Three of the infants required total parenteral nutrition (TPN) for an average of 49 days (range, 28-60) from diversion until the time of the definitive procedure. Only one patient did not receive TPN. Postoperatively, there were two complications-one wound infection and one hectic fever. The clinical results were good, with no soiling or stool incontinence and no constipation. CONCLUSION: The laparoscopic procedure for total aganglionosis or the extended form of Hirschprung's disease is safe, feasible, and reproducible. PMID- 11727139 TI - Endoscopic ligation for patients with active bleeding Mallory-Weiss tears. AB - BACKGROUND: Only a few patients with active nonesophageal variceal upper gastrointestinal bleeding have been treated with endoscopic ligation. To further address this issue, four patients with active bleeding Mallory-Weiss tears who underwent endoscopic band ligation are presented. PATIENTS AND METHODS: Endoscopic ligation was performed in four patients with a median age of 52 years (range, 40-93 years) after a diagnosis of active bleeding Mallory-Weiss tears (MWTs). A 45-year-old man with massive persistent upper gastrointestinal bleeding as a cause of a MWT underwent therapeutical endoscopic band ligation after an unsuccessful endoscopic injection trial. On the contrary, injection therapy should have been performed on a 93-year-old woman with multiple myeloma because of an actively bleeding MWT caused by the fibrotic tissue after an unsuccessful endoscopic ligation trial, although her other actively bleeding MWT lesion had been ligated successfully. RESULTS: After endoscopic ligation, all patients achieved complete hemostasis, and rebleeding did not occur. They were discharged without complications after a control endoscopy. CONCLUSIONS: Endoscopic ligation can be performed easily and without any complications such as perforation or delayed hemorrhage in patients with actively bleeding nonfibrotic MWTs. PMID- 11727140 TI - Laparoscopic spinal fusion of L4-L5 and L5-S1. AB - BACKGROUND: Patients with degenerative disc disease were treated by laparoscopic spondylodesis. Clinical outcome and quality of life were then analyzed in a retrospective study. METHODS: Thirty patients with a mean age of 43 years (range, 26-63) who suffered from instability of levels L4-L5 (n = 23), L5-S1 (n = 3), or both (n = 2) underwent surgery. Spondylodesis was achieved with two BAK cylinders. After 6 months, all patients completed a questionnaire designed to assess satisfaction with the operative outcome. RESULTS: After a median follow-up of 2.3 years (range, 0.9-3.5), 23 patients were free of pain or greatly improved. Complications included bleeding (n = 2), cage displacement (n = 1), retrograde ejaculation (n = 3), and postoperative ileus (n = 1). Good improvement was reported in daily activities and quality of life. CONCLUSION: Laparoscopic spondylodesis has a good clinical outcome with a low rate of morbidity. Notable improvements can be achieved in terms of daily activities and quality of life; however, these improvements are moderate in degree, and patient activity remains somewhat limited. PMID- 11727141 TI - Laparoscopic repair of poststernotomy subxiphoid epigastric hernia. AB - BACKGROUND: The repair of a subxyphoid hernia is a difficult procedure that nonetheless results in a high rate of recurrence. The laparoscopic approach is a promising new technique for more efficacious treatment of this condition. This is the first report in the English-language literature to describe the use of this approach for the correction of poststernotomy subxiphoideal hernia. METHODS: Information was retrieved from the patients' hospitalization and outpatient clinic files. Of 984 patients who had a median sternotomy, 10 developed a substernal subxiphoid epigastric hernia. These patients had all been treated laparoscopically using Gore-Tex mesh. RESULTS: Nine patients were admitted electively and one urgently. The fascial defect sizes were 4-15 cm (mean, 8.5) in length. Intraabdominal content was adherent to the hernia in six patients; in the other four cases, the defect was adhesion free. In four patients, an incidental surgical procedure was performed (three cholecystectomies and one inguinal hernia repair using the trans abdominal preperitoneal [TAPP] technique). The operations lasted 25-120 min (average, 55). No death occurred as a result of the operations, and none of the operations was converted to an open procedure. Three patients had minor postoperative complications. During 20-42 months of follow-up, one patient suffered a recurrence. CONCLUSIONS: Laparoscopic repair of a poststernotomy subxiphoideal epigastric hernia is feasible and has a low rate of minor complications. Our review of the literature indicates that this technique produces a better outcome than the conventional open repair. PMID- 11727142 TI - Laparoscopic surgery-induced changes in oxidative stress markers in human plasma. AB - BACKGROUND: The induction of the pneumoperitoneum increases intraabdominal pressure (IAP), causing splanchnic ischemia, whereas its deflation normalizes IAP and splanchnic blood flow. This procedure appears to represent an ischemia reperfusion model in humans. METHODS: Thirty laparoscopic cholecystectomies (LC) were performed in 30 patients with a mean age of 54.6 +/- 15.6 years. A group of 20 patients mean age, 57.3 +/- 9.65 who underwent open cholecystectomy (OC) was also studied. Vein plasma levels of thiobarbituric acid-reactive substances (TBARS), a marker of free radical production; plasma total antioxidant status (TAS); and uric acid (UA) levels were measured preoperatively, 5 min after deflation of the pneumoperitoneum or at the end of operation, and 24 h postoperatively. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TBL) levels were measured preoperatively and 24 h after the operation. RESULTS: In the LC group, significant elevations in the concentration of TBARS were observed in the early postoperative measurements in comparison with the preoperative measurements. TAS and UA levels were decreased significantly 24 h postoperatively compared to preoperative levels. The postoperative levels of AST, ALT, and TBL increased significantly in comparison with the preoperative levels. In the OC group, no alterations in the concentration of TBARS were observed in the postoperative period. The other parameters had changes similar to those recorded for the LC group. CONCLUSIONS: Free radical-induced lipid peroxidation associated with a decrease in plasma antioxidant capacity and UA levels as well as altered hepatic function is observed after deflation of the pneumoperitoneum. These results suggest that free radicals are generated at the end of a laparoscopic procedure, possibly as a result of an ischemia-reperfusion phenomenon induced by the inflation and deflation of the pneumoperitoneum. PMID- 11727143 TI - Laser-induced thermotherapy: intra- and extralesionary recurrence after incomplete destruction of experimental liver metastasis. AB - PURPOSE: The aim of this study was to determine the energy (J/mm3 tumor volume) and temperature required for a complete laser-induced thermotherapy (LITT) of experimental liver tumors, and to find out causes and areas of local recurrence followed by incomplete treatment. METHODS: In VX-2 tumor-bearing rabbits LITT was performed using neodymium:yttrium-aluminum-garnet (Nd:YAG) laser (1064 nm) with a diffuser-tip applicator and a temperature feedback system. The animals were randomized into four groups (n = 20) that differed in the target temperature at the tumor border as follows: 45 degrees C, 50 degrees C, 55 degrees C and 60 degrees C. The target temperature was held for 10 min constant. Histologic examination (hematoxylin and eosin [H and E], nicotinamide adenine dinucleotide phosphate [NADPH]-dehydrogenase) was performed at 0 h, 24 h, 96 h, and 14 days after LITT. RESULTS: The pretreatment tumor volume of 2191 +/- 61 mm3 was the same for all groups (p > 0.05). Energy up to target temperature and total energy required, lesion size, and the rate of incomplete tumor ablation (recurrences) are listed below (ap < 0.05, Kruskal-Wallis test). Histologically, two forms of local recurrences could be differentiated intralesionary and extralesionary. CONCLUSIONS: To achieve complete in situ ablation under the given conditions, it is necessary to apply laser energy of 3 J/mm3 tumor volume. A minimum temperature of 60 degrees C on the tumor border presumed an application of 10 min. Recurrence was found outside the coagulation zone (extralesionary) and in high vascularized areas within the coagulation zone (intralesionary). PMID- 11727144 TI - Hemodynamic responses to intrapleural insufflation with hemipulmonary collapse. AB - BACKGROUND: The hemodynamic effects of carbon dioxide (CO2) insufflation with hemipulmonary collapse were studied in consecutive thoracoscopic harvests of the left or right internal mammary artery (IMA), which were used for video-assisted coronary artery bypass grafting. METHODS: Thirty-eight patients (30 male, eight female) with a mean age of 69.5 +/- 11.5 years were selected, and 33 left and five right IMA were harvested thoracoscopically. After hemipulmonary collapse was established by single-lung ventilation, low-flow (2-3 L/min) CO2 was delivered at a constant intrapleural pressure of 8-10 mmHg. Using electrocardiography, a radial arterial catheter, a Swan-Ganz catheter, and transesophageal echocardiography, we obtained values for seven hemodynamic variables. Baseline data were collected during bilateral lung ventilation. Each variable was then measured during hemipulmonary collapse and insufflation. The significance of any changes was established with Student's t-test after correcting for baseline differences. RESULTS: Insufflation facilitated IMA harvest by expanding the pleural space between the anterior chest wall and heart. Mean insufflation times were 40.8 +/- 12.2 min on the left and 33.5 +/- 8.5 min on the right. Significant increases from the baseline values were observed in the mean central venous pressure (L:4.7-9.0 mmHg, R: 5.1-14.0 mmHg, p < 0.05), the pulmonary arterial pressure (L: 11.3-17.3 mmHg, R: 12.1-19.9 mmHg, p < 0.05), and the pulmonary capillary wedge pressure (L: 7.2-10.5 mmHg, R: 6.5-10.0 mmHg, p < 0.05). On the right, but not on the left, slight decreases were noted in the mean arterial pressure and cardiac index (71.3-62.6 mmHg, 2.01-1.76 L min-1 m-2, p < 0.05). CONCLUSIONS: The hemodynamic effect resulting from one-lung collapse plus low flow CO2 insufflation at 8-10 mmHg for 30-40 min is mild in both hemithoraces, although the impact is greater on the right. PMID- 11727145 TI - Clonidine provides opioid-sparing effect, stable hemodynamics, and renal integrity during laparoscopic cholecystectomy. AB - BACKGROUND: Carbon dioxide pneumoperitoneum causes a hemodynamic stress response and decreases urine output because of an activated renin-angiotensin-aldosterone system (RAAS). Clonidine is a potent antihypertensive drug that suppresses RAAS. METHODS: The effects of clonidine 4.5 mg/kg or saline on hemodynamics, neuroendocrine response, and renal parameters were compared in 30 healthy patients undergoing laparoscopic cholecystectomy. RESULTS: Heart rate, arterial blood pressures, and plasma renin activity were lower during and after pneumoperitoneum in patients with clonidine. There were no differences in urine output, urine oxygen tension (reflecting medullary perfusion), or antidiuretic hormone between the groups. N-acetyl-b-D-glucosaminidase, a marker of proximal tubular damage, was minimally elevated after clonidine. CONCLUSIONS: Clonidine enabled stable hemodynamics and prevented activation of RAAS seen as unchanged plasma renin activity. Clonidine may be beneficial during laparoscopy in patients with hypertension, cardiovascular, and/or renal diseases. PMID- 11727146 TI - Laparoscopic cholecystectomy in morbidly obese patients. AB - BACKGROUND: Morbid obesity is generally regarded as a risk factor for laparoscopic cholecystectomy due to increases in operative time, morbidity, and conversion rate to open cholecystectomy. The aim of this study was to evaluate the feasibility and outcome of laparoscopic cholecystectomy (LC) in morbidly obese patients. METHODS: A total of 864 consecutive patients underwent LC at our institution between 1990 and 1997. This series represents a continuing policy of LC for all comers. Data were collected prospectively. There were 659 nonobese (NO: BMI 40 kg/m2). Laparoscopic bile duct exploration was performed in 28 (4.2%), nine (4.8%), and one (5.9%) patients, respectively. RESULTS: Obesity and morbid obesity were associated with trends toward an increased conversion rate (2.3% NO; 4.3% OB; 5.9% MO), a longer operative time (median, 80, 85, and 107 mins, respectively), greater postoperative morbidity (4.7%, 5.9%, and 11.8%, respectively), and a reduced ability to obtain cholangiography (86.1%, 80.1%, and 71.4%, respectively). None of these differences, however, were statistically significant (c2 test, p > 0.05). Postoperative hospital stay for LC was similar for all three groups (median, 1 day). CONCLUSION: LC in morbidly obese patients is a safe procedure, but it may be associated with increased operative difficulty and morbidity, as compared with nonobese and obese patients. PMID- 11727147 TI - Fiberoptic ductoscopy for breast cancer patients with nipple discharge. AB - BACKGROUND: Breast cancer and precancer are thought to originate in the lining of the milk duct, but until recently, we have not had direct access to this area other than in tissue removed blindly by core biopsy or fine-needle aspiration. Fiberoptic ductoscopy (FDS) is an emerging technique that allows direct visual access of the ductal system of the breast through nipple orifice cannulation and exploration. To date, this technique has been used only in pilot studies. Previously, we have demonstrated that fiberoptic ductoscopy in patients with and without nipple discharge is a safe and effective means of visualizing the intraductal lesion. When combined with cytology, it is a screening technique that has high predictive value. METHODS: We applied ductoscopy to 415 women with nipple discharge with the specific intent of detecting those patients with nipple discharge who had intraductal carcinoma (DCIS) as the basis of their discharge. RESULTS: In this cohort of patients, ductoscopy was successful in visualizing an intraductal lesion in 166 patients (40%). In these cases, ductal lavage following ductoscopy increased the yield of cytologically interpretable ductal epithelial cells 100-fold compared to discharge fluid alone. In the majority of these patients, FDS examination detected lesions that had the appearance of typical papillomas. However, in 10 patients, the intraductal lesion exhibited one of several atypical features, including bleeding, circumferential obstruction, and gross fungating projections. In eight of these patients, the subsequent histopathology turned out to be DCIS. In two of these eight patients, endoscopic biopsy revealed cytologically malignant cells; in two others, ductal lavage (washings) revealed cytologically malignant cells. In three additional patients, although FDS examination uncovered a typical papilloma that was not biopsied, ductal lavage (washings) revealed cytologically malignant cells. On surgical pathology review of the extirpated lesions, all 11 patients were subsequently shown to have DCIS. Of these 11 cases of DCIS that were initially detected with a combination of FDS and ductal lavage cytology, six were completely negative on mammogram and physical exam. CONCLUSION: Although nipple discharge is an unusual presentation for DCIS, in patients with nipple discharge, FDS with ductal lavage cytology is a useful technique for diagnosing DCIS prior to definitive surgery. PMID- 11727148 TI - Description of two new human ovarian carcinoma models in nude rats suitable for laparoscopic experimentation. AB - BACKGROUND: Experimental laparoscopic trials require relevant models of ovarian carcinomatosis. METHODS: Female nude rats were inoculated intraperitoneally either with the IGR-OV1 or the NIH:OVCAR-3 human adenocarcinoma cell lines. Serial clinical checks and sacrifices were used to evaluate the rates of tumor take, survival, and patterns of tumor spread. Finally, laparoscopies with various pneumoperitoneum pressures were performed to verify the "surgical" relevancy of out models. The learning curve was measured. RESULTS: The best results were obtained when twenty-seven 106 IGR-OV1 cells and thirty-six 106 NIH:OVCAR-3 cells were injected in 28-day-old rats. The IGR-OV1 model provided a mean survival of 17.8 days (range, 13-22 days), with a high take rate (94%). The NIH:OVCAR-3 model resulted in a longer mean survival (59 days; range, 49-77) and also a high take rate (83%). The two models differed in their patterns of tumor spread: solid bulky omental metastasis having a diffuse microscopic peritoneal carcinomatosis with the IGR-OV1 line (the weight of the omental cake correlated significantly with the stage of development) and diffuse macroscopic peritoneal carcinomatosis having no large solid tumor, but visceral and paraaortic metastases, with the NIH:OVCAR-3 line. In both models, CA125 was high. Anesthesia could be performed and repeated in healthy and tumor-bearing rats. Laparoscopy was feasible, with pneumoperitoneum pressures as high as 8 mmHg lasting 1 h. Laparoscopy provided a reliable evaluation of the tumor spread into the peritoneal cavity. The plateau of the learning curve was soon obtained for take rate and survival after laparoscopy. CONCLUSION: We report two new human ovarian carcinoma xenografts in nude rats suitable for laparoscopy. The IGR-OV1 model mimics an advanced stage of the disease, and the NIH:OVCAR-3 model presents an earlier stage. These two models appear useful for experiments involving laparoscopy. PMID- 11727149 TI - The role of laparoscopy in the management of common bile duct obstruction in children. AB - BACKGROUND: Laparoscopic common bile duct exploration is commonplace in adults; however, this procedure is not often performed in children. The goal of this study was to evaluate the results of laparoscopic common bile duct exploration in children. METHODS: Of 50 patients undergoing laparoscopic cholecystectomy, six patients (12%) had obstructing lesions of the common bile duct (CBD). Five children underwent laparoscopic common bile duct exploration, and one child had a preoperative endoscopic sphincterotomy and stone removal. RESULTS: The mean age at laparoscopic CBD exploration was 11.6 years (range, 5-16). The obstructing lesion was visualized by intraoperative cholangiography in all five patients. The mean operative time for laparoscopic cholecystectomy along with CBD exploration was 215 min (range, 160-282). The transcystic laparoscopic CBD exploration was performed using a 7-Fr, multichannel rigid, or 10-Fr flexible fiberoptic cystoscope. The stones were either pushed into the duodenum with the scope or extracted through the cystic duct using a 3-Fr Segura basket. In one patient, a candidial ball disintegrated during an attempt to remove it with the basket. A repeat cholangiogram at the end of each procedure showed an anatomically normal CBD with free flow of contrast into the duodenum. All patients enjoyed a quick recovery. They were started on a regular diet on the same day of surgery and discharged on the 1st or 2nd postoperative day. One patient with sickle cell disease developed a pulmonary infarction and required 5 additional days of hospitalization. One patient developed recurrent choledocholithiasis 6 months after laparoscopic exploration and was treated successfully with endoscopic sphincterotomy and stone extraction. CONCLUSIONS: Laparoscopic CBD exploration can be performed safely at the time of the cholecystectomy in children. Endoscopic sphincterotomy before cholecystectomy is not necessary. We recommend laparoscopic CBD exploration for obstructing lesions of the CBD. Endoscopic sphincterotomy should be reserved for recurrent lesions of the CBD after laparoscopic cholecystectomy. PMID- 11727150 TI - Laparoscopic vs open adrenalectomy for benign adrenal neoplasm. AB - BACKGROUND: The aim of this study was to compare the outcome of laparoscopic adrenalectomy (LA) performed for benign adrenal neoplasm to the open procedure in a similar group of patients. METHODS: All consecutive patients who underwent LA between June 1996 and February 1999 were evaluated. Data analysis included patient's age and gender, indication for surgery, histological diagnosis, size of specimen, comorbid conditions, length of stay and ileus, postoperative narcotic consumption, and time to return to normal activity. The results were compared retrospectively to a well-matched group of patients who underwent an open adrenalectomy (OA). RESULTS: Twenty-eight LA were performed in 24 patients for the following disorders: adrenocortical adenoma, 16 (four Cushing's syndrome, 12 Conn's syndrome); pheochromocytoma, 10; and nonfunctioning tumor, two. These cases were compared with a well-matched group of 28 patients who underwent OA in the same department. There were two conversions to open surgery (7%) in the laparoscopic group and no deaths in either group. Of all the evaluated parameters, the following statistically significant differences between the two groups were noted: The mean operative time was longer in the LA group (188 vs 139 min, p < 0.001.); however, this became insignificant in the last 10 cases of LA, when the mean length of surgery was reduced to 130 min. The overall morbidity was lower in the LA group (16% vs 39%, p = 0.05), as was the mean time to tolerate a regular diet (2 vs 3.9 days), mean meperidine consumption (mg) (109 vs 209), mean length of stay (4 vs 7.5 days), and mean time to return to normal activity (2.2 vs 5.2 weeks), (p < 0.001 for all). CONCLUSION: LA for benign adrenal disorders is a safe procedure that is associated with significantly lower morbidity, shorter ileus and hospitalization, reduced postoperative pain, and a faster return to normal activity than the open procedure. PMID- 11727151 TI - Laparoscopic repair of Bochdalek's hernia with gastric volvulus. AB - Bochdalek's hernia is the most common congenital diaphragmatic hernia in newborns, but it is an uncommon clinical feature in adults. In most cases, there are no symptoms or only minor gastrointestinal problems. Rarely, it may manifest as an acute abdomen due to viscus strangulation, requiring emergency surgery. We report the case of a 52-year-old man with herniation of the colon and a volvulated stomach through a Bochdalek's hernia. The patient was operated on via a laparoscopic approach and had an uneventful recovery. We recommend the laparoscopic approach as an alternative to open surgery in cases of noncomplicated Bochdalek's hernia. PMID- 11727152 TI - Laparoscopic-assisted resection of bleeding jejunal leiomyoma. AB - Laparoscopic-assisted resection has shown remarkable improvements in the treatment of small bowel diseases, notably the leiomyomas. This case report documents the successful removal of a bleeding jejunal leiomyoma with the aid of laparoscopy. A 51-year-old man was admitted to the hospital twice within 3 months with melena. On the first admission, upper and lower gastrointestinal endoscopy were negative, and small bowel enema was inconclusive. On the second admission, Technetium-99 Red Blood Cells (Tm-99 RBC) scan showed dye extravasation (interpreted as from the left colon). Subsequent colonoscopy was normal, as was a barium enema. An elective superior mesenteric angiography revealed a benign looking jejunal leiomyoma. Subsequent CT (intravenous contrast scan) revealed a 4.6 x 3.5 cm mass with neither extraluminal infiltration nor enlarged lymph nodes. The patient underwent successful laparoscopic-assisted resection of the leiomyoma and enjoyed an uneventful postoperative recovery. Subsequent histopathology confirmed the diagnosis. The diagnosis of small bowel leiomyoma is generally difficult because the conventional radiographic methods are unhelpful. These tumors are therefore best detected preoperatively with superior mesenteric angiography. Repeat of the noninvasive tests is strongly indicated if they are initially negative. Once discovered, small bowel leiomyoma should be resected to avoid catastrophic complications. Thereafter, it has an excellent prognosis. In this context, minimal-access surgery is a safe and approachable method to deal with such a problem. PMID- 11727153 TI - Radiological treatment of retained bile duct stones following recent surgery using glucagons. AB - BACKGROUND: Retained common bile duct (CBD) stones pose an occasional problem following ductal exploration, in spite of completion cholangiography or choledochoscopy. We present a method for treating retained stones in the Radiology Department by biliary lavage via a transcystic tube (TCT) or a T-tube, after intravenous administration of glucagon. METHODS: A TCT or T-tube is inserted following CBD exploration for multiple intrahepatic stones or when stones are fragmented to facilitate removal or flushing into the duodenum. A tube cholangiogram is performed on the 1st postoperative day. If any retained stones are encountered, 1 mg glucagon is administered intravenously and saline irrigation through the tube is done under fluoroscopic control, allowing the stone to pass to the duodenum. The cholangiogram is repeated 10-14 days later, before removing the tube. RESULTS: In case 1, transcystic CBD exploration was performed. Two stones were crushed and flushed into the duodenum. TCT cholangiography the following day. showed a 5-6-mm fragment causing complete obstruction. Following the use of glucagon and irrigation, the stone was observed passing into the duodenum, causing a brief mild episode of pain. In case 2, laparoscopic choledochotomy was performed to remove seven large stones. Completion choledochoscopy was satisfactory. T-tube cholangiography identified a small stone in the CBD, which was cleared with the help of glucagon. CONCLUSION: The current standard treatment for retained stones is endoscopic sphincterotomy. This is associated with morbidity, mortality, and significant additional cost. This new technique is a simple and safe alternative for retained CBD stones, most of which as small stones or fragments. Because glucagon causes intense relaxation of the sphincter of Oddi, the procedure should not take much longer than a routine tube cholangiogram. The safety of glucagon makes it possible to repeat the procedure if necessary. PMID- 11727155 TI - Fatal pulmonary thromboembolism after laparoscopic cholecystectomy. AB - A 67-year-old woman presented with severe cardiopulmonary insufficiency 17 days after an uneventful laparoscopic cholecystectomy. Pulmonary thromboembolism was demonstrated by transthoracic echocardiogram and later confirmed at surgery. With the aid of a cardiopulmonary bypass, a thrombectomy of the right atrium and the pulmonary artery was accomplished. The patient could not be weaned off cardiopulmonary bypass and ultimately died. We therefore recommend antithromboembolism therapy with low-molecular-weight heparin in selected cases of laparoscopic cholecystectomy. PMID- 11727156 TI - Pneumatosis cystoides intestinalis as a complication of celiac disease. AB - Pneumatosis cystoides intestinalis (PCI) is an uncommon but important condition characterized by the presence of gas-filled cysts in the submucosa and subserosa of the gastrointestinal tract. PCI has been associated with several clinical settings. We report a case where PCI developed in a patient with known celiac disease. To our knowledge, this type of coincidence has been described in seven prior cases. Since PCI often results in pneumoperitoneum through rupture of the cysts, it is important to differentiate the benign form of pneumoperitoneum, in which no intervention is indicated, from the life-threatening form with intestinal infarction and/or perforation, in which immediate surgery is mandatory. Differentiating between them can be difficult; this explains the high rate of negative laparotomy, which, in the past, occurred in 27% of cases. By performing diagnostic laparoscopy, we were able to establish the diagnosis of PCI and exclude intestinal perforation or infarction, thus permitting the patient to avoid an unnecessary laparotomy. Although diagnostic laparoscopy is not routinely indicated as a diagnostic tool for the detection of PCI, it proved to be a safe and accurate method to differentiate between the benign form of PCI and the life threatening form. Whenever this differentiation has to be made, we recommend that diagnostic laparoscopy be performed first rather than primary laparotomy. Herein, the laparoscopic features of PCI are described and the coincidence of PCI with celiac disease is reviewed. PMID- 11727154 TI - Totally endoscopic Nissen fundoplication with a robotic system in a child. AB - The Da Vinci robot device represents the latest advance in laparoendoscopic surgery. We report the case of an endoscopic Nissen fundoplication performed with the aid of a telemanipulation robot system in a 10-year-old girl. To our knowledge, this is the first such case to be reported. In addition to standard monitoring, we used invasive blood pressure monitoring during the 282-min period of general anesthesia. Arterial blood gas samples were analyzed at short intervals. During surgery, which included a 172-min period of intraperitoneal insufflation of carbon dioxide (CO2), no significant changes were observed in PH, arterial oxygen pressure (PaO2), arterial carbon dioxide pressure (PaCO2), heart rate, or mean arterial pressure. Body temperature was maintained with an external warming blanket. Total intravenous anesthesia with continously administered propofol, remifentanil, and mivacurium for continous muscle relaxation allowed extubation immediately after skin closure. The girl was discharged from hospital on postoperative day 6. Robot-assisted techniques have the potential to significantly improve the performance of laparoendoscopic surgery. However, despite our encouraging first results, the potential risks of robot-assisted surgery have not yet been definitively established. Therefore, patients submitted to this type of procedure require intensive and even invasive monitoring. PMID- 11727157 TI - Adjustable gastric banding in a patient with sarcoidosis. AB - The use of silicone in patients with immune-compromising diseases is controversial because of fears that there may be inflammatory reactions against the material. We report the case of a patient who suffered from cutaneous sarcoidosis with bihilar lymphadenopathy and obesity (110 kg; BMI, 38 kg/m2) that was exaggerated by cortisone therapy. The patient underwent adjustable gastric banding because of aggravating comorbid hypertension and hyperlipidemia. Six months postoperatively, she had achieved an excess weight loss of 33%. During this period, she had a relapse of scar sarcoidosis of the right elbow and bilateral hilar lyphadenopathy. Although she was treated with a double dose of cortisone (8 mg daily), the scars from the laparoscopy as well as those in the gastric region adjacent to the band remained unaffected. We therefore believe that preexisting immune-compromising diseases are not an absolute contraindication against gastric banding, particularly since our patient experienced a notable improvement in her quality of life. PMID- 11727158 TI - Endoscopic thyroidectomy by the axillary approach. AB - Neck surgery has recently become one of the newest fields for the application of endoscopic surgery because the resultant scar is small and inconspicuous. Still, some patients feel even a small scar on the neck is not cosmetically acceptable. We therefore have developed a new technique of endoscopic thyroidectomy by the axillary approach that leaves no scar on the neck at all. When this method is used, the small scar in the axilla is completely covered by the patient's arm in a natural manner. The cosmetic result is excellent, and sensory loss in the neck is negligible because the area of surgical dissection is small. We believe that endoscopic thyroid surgery by the axillary approach will find a role in the treatment of thyroid disease. PMID- 11727159 TI - Transthoracic endoscopic sympathectomy. PMID- 11727160 TI - [Highly resistant pathogens in patients with diabetic foot syndrome with special reference to methicillin-resistant Staphylococcus aureus infections]. AB - BACKGROUND AND OBJECTIVE: Patients with diabetic foot infection (DFI) have a high rate of infection, up to 40%, with methicillin-resistant Staph. aureus (MRSA). Having noticed a definite increase of such patients in our special unit, we initiated a drastic change of hygienic measures and here report the results. PATIENTS AND METHODS: 788 patients with DFI (mean age 67.3 [32-90] years, 62% males) were admitted between 1.1.1999 and 31.7.2000. Before 31.7.1999, the following hygienic measures had been in place: cohort isolation, protective closing, implementation of general hygienic rules. Since 1.8.1999, modified measures have been undertaken: primary single-patient isolation, concentration in one ward of all patients with MRSA, medical care only by trained personnel, admission of patients only after microbiological results were known or primary solitary isolation. Algorithms were used for the transmission of all necessary information. RESULTS: MRSA was demonstrated in 64 patients. The number of infections during the hospital stay, before and after the change of hygienic measures were 9 (27%) and 2 patients (8%), respectively. The sites of MRSA colonisation and proven eradication were: nasopharynx only, 3 with 67% eradication; MRSA in a wound, 25 with 28% eradication. In comparison to the yearly statistic on wound healing in DFI 1999 (n=613) the following results are shown (patients with MRSA in brackets): healing rate with conservative treatment 61.5% (20%), minor-amputation 30.5% (52%), major-amputation 4.5% (22%), death 3.5% (6%). CONCLUSIONS: The rate of new infections were dramatically reduced by changing the hygienic measures. The rate of successful sanitation was unsatisfactoy. Patients with MRSA showed markedly poorer treatment results in respect to wound healing. PMID- 11727161 TI - [Renal oxalosis with renal failure after infusion of xylitol]. AB - HISTORY AND FINDINGS: Ten days after a complication-free operation for early gastric cancer a 61-year old man had a seizure and acute renal failure. INVESTIGATIONS: A percutaneous renal biopsy was performed to find the cause of the renal failure, which at times required several sessions of haemodialysis. TREATMENT AND COURSE: The seizure was thought to have been due to cerebrovascular deposits of oxalate crystals (renocerebral oxalosis). Parenteral xylitol had been given postoperatively, as a glucose substitute (total dose 1560g), together with other routine drugs. Primary oxalosis and other causes of secondary oxalosis had been excluded. CONCLUSION: Xylitol should not be used parenterally as a glucose substitute (banned in the USA). PMID- 11727162 TI - [Early pain reduction in the treatment of spasticity after a single injection of botulinum A toxin]. AB - HISTORY, ADMISSION FINDINGS AND DIAGNOSIS: After stem-cell transplantation a 45 year-old woman (case 1) had an attack of general hypoxia requiring resuscitation. She then developed a quadriplegia and spasticity of all limbs notably of the right arm and a severe pain syndrome which had to be treated by oral and intravenous analgesics. Immobilisation and secondary complications aggravated the already difficult situation. In the 2nd case a 66-year-old woman was admitted to our outpatient clinic with long-standing left-sided spastic hemiparesis after territorial infarction of the right middle cerebral artery. Beside the spasticity she also suffered from a distinct pain syndrome which did not respond to any oral analgesics. TREATMENT AND COURSE: For the treatment of the main symptoms, both patients received intramuscular injections of 1000 MU botulinum toxin A (Dysport(R) Ipsen Pharma). Astonishingly, both patients experienced pain relief the next day, whereas spasticity started to respond only 5-6 days later. CONCLUSIONS: In our experience pain relief after botulinum toxin A injections occurs not only due to reduced muscle hyperactivity, especially when such a temporal dissociation between pain relief and muscle relaxation appears as in the two cases reported above. Rather, we believe that botulinum toxin A interferes with the release of other neurotransmitters e. g. substance P (SP) and calcitonine-gene-related-peptide (CGRP) having a key function in the nociceptive cascade. PMID- 11727163 TI - [New methods for demonstrating nucleic acids]. PMID- 11727164 TI - [Pulmonary amyloidosis]. PMID- 11727166 TI - [Radon therapy in Bechterew disease. Benefits and risk factors]. PMID- 11727165 TI - [The diagnosis of pneumonia in daily practice: how many diagnostic measures are essential?]. PMID- 11727167 TI - [Ciguatera: clinical relevance of a marine neurotoxin]. PMID- 11727169 TI - [Percutaneous revascularization of long-standing chronic occlusion of the superficial femoral artery]. PMID- 11727170 TI - [Arteriovenous fistula in Osler's disease]. PMID- 11727171 TI - Noncontraceptive benefits and therapeutic uses of the oral contraceptive pill. AB - The oral contraceptive pill is one of the most extensively studied medications ever prescribed. The health benefits are numerous and outweigh the risks of their use. Definitive evidence exists for protection against ovarian and endometrial cancers, benign breast disease, pelvic inflammatory disease requiring hospitalization, ectopic pregnancy, and iron-deficiency anemia. It has also been suggested that oral contraceptives may provide a benefit on bone mineral density, uterine fibroids, toxic shock syndrome, and colorectal cancer. Minimal supportive evidence exists for oral contraceptives protecting against the development of functional ovarian cysts and rheumatoid arthritis. Treatment of medical disorders with oral contraceptives is an "off-label" practice. Dysmenorrhea, irregular or excessive bleeding, acne, hirsutism, and endometriosis-associated pain are common targets for oral contraceptive therapy. Most patients are unaware of these health benefits and therapeutic uses of oral contraceptives, and they tend to overestimate their risk. Counseling and education are necessary to help women make well-informed health-care decisions and improve compliance. PMID- 11727172 TI - The risks of oral contraceptive pills. AB - Oral contraceptive pills have been associated with increased risk for myocardial infarction, stroke, and venous thromboembolism. Studies have been published recently that suggest that these risks are minimal in appropriately chosen low risk women. Stroke is a very uncommon event in childbearing women, occurring in approximately 11 per 100,000 women over 1 year. Thus, even a doubling of this risk with oral contraceptive pills would have minimal effect on attributable risk. The estimated risk of myocardial infarction associated with oral contraceptive pill use in nonsmokers is 3 per million women over 1 year. The estimated risk of venous thromboembolism attributable to oral contraceptive pills is less than 3 per 10,000 women per year. Additionally, the literature suggests that there may be an increased risk of breast cancer associated with long-term oral contraceptive pill use in women under the age of 35. However, because the incidence of breast cancer is so low in this population, the attributable risk of breast cancer from birth control pill use is small. PMID- 11727173 TI - Successful use of oral contraceptives. AB - Typical failure rates for oral contraceptives remain much higher than ideal failure rates. Patients can cite the pill's perceived risks more readily than its benefits, and many women use oral contraceptives inconsistently or discontinue them entirely without a medical reason. Successful use of oral contraceptives mandates that we rethink our roles as providers. Medical barriers, such as mandatory pelvic examinations, impede clients' access to services and require reevaluation. Efforts must also focus on thorough, individualized counseling. Because compliance poses difficulties for many women and noncompliance results in reduced efficacy, counseling must address content points relevant to proper, consistent use. Additionally, as unexpected side effects often cause frustration and method discontinuation, counseling should anticipate their occurrence. Adolescents present additional challenges. However, with careful attention to concerns relevant to teenage women and consideration of obstacles to compliance, oral contraceptives can indeed be used successfully in this group. Regardless of age, all clients should receive information regarding emergency contraception, and instructions for pill use should be individualized to meet the needs of each patient. PMID- 11727174 TI - Emergency contraception. AB - Emergency contraceptives are methods that prevent pregnancy when used shortly after unprotected sex. Three different emergency contraceptive methods are safe, simple, and widely available in the United States. These are: (1) ordinary combined oral contraceptives containing ethinyl estradiol and levonorgestrel taken in a higher dose for a short period of time and started within a few days after unprotected intercourse; (2) levonorgestrel-only tablets used similarly; and (3) copper-bearing intrauterine devices inserted within approximately 1 week after unprotected intercourse. Emergency contraceptive use is best known for women who have been raped, but the methods are also appropriate for women who have experienced condom breaks, women who did not use any method because they were not planning on having sex, or women who had unprotected intercourse for any other reason. Unfortunately, few women know about emergency contraceptives, and few clinicians think to inform their patients routinely about the option. A nationwide toll-free hotline (1-888-NOT-2-LATE) and a website (http://not-2 late.com) can help women learn about these options. Sharing "family planning's best-kept secret" widely with women could prevent as many as a million unwanted pregnancies annually in the United States. PMID- 11727175 TI - Current options for injectable contraception in the United States. AB - Two injectable forms of hormonal contraception, depot medroxyprogesterone acetate (DMPA, Depo-Provera) and medroxyprogesterone acetate/estradiol cypionate (MPA/E(2)C, Lunelle), are now available to American women. Both formulations have demonstrated high degrees of efficacy, safety, and ease of use in international and U.S. trials. Data on DMPA have shown a number of noncontraceptive and therapeutic benefits, the most prominent of which is an 80% reduction in the risk of endometrial cancer. Although such benefits are less documented for MPA/E(2)C, they are expected to be similar to those seen with DMPA and oral contraceptives. Minor side effects of both formulations include menstrual irregularities in the early months of treatment and amenorrhea with DMPA. Patient counseling about the potential for these side effects, as well as possible risks, is important to long term successful use of these contraceptive methods. PMID- 11727176 TI - Implant contraception. AB - The experience of 6 million Norplant users has led to several more advanced implants. Implanon is a single-rod implant system containing a low androgenic progestin and requires 1 to 2 minutes for insertion and removal. Like other implants, Implanon prevents pregnancy by changing the character of the cervical mucus and interfering with luteal function. Unlike Norplant, though, Implanon is designed to prevent ovulation for the full duration of use. Implant contraception has several advantages over other types of contraception including high efficacy, minimal required maintenance, absence of estrogen, and rapid return of fertility after discontinuation. Implants can be a good choice for adolescents; women with hypertension, diabetes, anemia, endometriosis, or other medical problems; and women who are breast-feeding. Irregular bleeding is the most common adverse effect of implants and can be treated with several medication regimens. Preinsertion counseling, however, is the most important factor in ensuring satisfaction with implants. Unfortunately, no implant system is currently available in the United States since August 2000, but Implanon is expected to reach the U.S. market within the next 2 years. PMID- 11727177 TI - Progestin-releasing intrauterine systems. AB - The levonorgestrel-releasing intrauterine system (LNG IUS) is an effective method for contraception. It has a strong antiproliferative action on the endometrium. The endometrium is transformed under the influence of local levonorgestrel and becomes unresponsive to ovarian estrogens. This process is associated with progressive reduction of menstrual blood loss and menstrual duration. Scanty and irregular bleeding and/or spotting is usual during the first 3 to 4 months. The reduction of menstrual blood loss continues and after the first 9 months many women have no bleeding at all. However, they have normal ovarian function. The absence of bleeding is a result of the local antiproliferative action of the LNG IUS on the endometrium, which is also responsible for many health benefits during the use of this method. As with oral contraceptives, the risk of pelvic inflammatory disease is reduced, because of reduced menstrual blood loss, endometrial suppression, and thickening of the cervical mucus. There are some steroidal side effects: mood changes, oily skin, and acne. Weight increase is similar to that associated with copper intrauterine devices: 500 g per year over 5 years. Users should be told that the LNG IUS does not prevent sexually transmitted infection, and therefore women at risk should also use condoms for their protection. PMID- 11727178 TI - Therapeutic use of the LNG IUS, and counseling. AB - The intrauterine release of levonorgestrel (LNG) alters the function of the endometrium. This phenomenon offers special health benefits for users and a manner of local intrauterine therapy. In this review we discuss use of LNG releasing intrauterine system (LNG IUS) in treatment and prevention of anemia and in the therapy of menorrhagia and dysmenorrhea. The antiproliferative effect of the LNG IUS on the endometrium offers targeted therapy against the proliferative action of estrogen on the endometrium during hormone replacement therapy. The LNG IUS as an alternative to sterilization is also discussed. There are also promising therapeutic fields such as use of the LNG IUS in the treatment of endometriosis and in the protection of endometrium exposed to tamoxifen during breast cancer treatment. Special attention is paid to counseling because successful use of the LNG IUS requires good training of the providers and extensive counseling of users. PMID- 11727179 TI - Transdermal contraception. AB - This review summarizes the clinical studies involving the once-weekly Ortho Evra/Evra contraceptive patch. The patch delivers norelgestromin (NGMN), 150 microg, and ethinyl estradiol (EE), 20 microg, daily to the systemic circulation. The contraceptive patch provided ovulation suppression and cycle control similar to that of oral norgestimate 250 microg/EE 35 microg, significantly decreased mean maximum follicular diameter following a 3-day intentional delayed dosing phase when compared with oral levonorgestrel (LNG) 50/75/125 microg/EE 30/40/30 micorg and oral LNG 100 microg/EE 20 microg, and was as effective as oral LNG 50/75/125 microg/EE 30/40/30 microg and oral desogestrel 150 microg/EE 20 microg in altering cervical mucus composition (i.e., creating a scanty, viscous consistency). The contraceptive patch provided efficacy, cycle control, and safety comparable to that seen with oral LNG 50/75/125 microg/EE 30/40/30 microg, but women were able to correctly follow the weekly dosing regimen significantly more often than the daily oral contraceptive dosing regimen. Less than 2% of patches were replaced because of complete detachment in these trials. The patch was not associated with phototoxicity or photoallergy. The contraceptive patch, the only noninvasive, weekly birth control method that a woman can self administer, will be a valuable addition to current contraceptive options. PMID- 11727181 TI - [Surgery in Eastern Europe]. PMID- 11727180 TI - Contraceptive vaginal rings. AB - Contraceptive vaginal rings (CVRs) contain sex steroids that diffuse through a plastic polymer ring at a constant rate and are absorbed directly through the vaginal epithelium into the systemic circulation. This delivery system provides many advantages over oral contraceptives (OCs), including avoidance of the first pass effect through the liver, constant serum steroid levels, longer duration of use, and greater bioavailability of the hormones. CVRs containing progestin only are designed for continuous use for 3 to 6 months. Those containing progesterone alone are indicated for use in women who are breastfeeding. Large clinical trials of progestin-containing CVRs demonstrated good efficacy and safety of the CVR, with continuation rates similar to that of OCs. CVRs containing a combination of estrogen and progestin are designed to be used for 1 to 12 months in a cyclic manner similar to OCs, with withdrawal bleeding in the fourth week of each cycle. In clinical trials these CVRs have typical use efficacies similar to OCs, with an acceptable pattern of bleeding. PMID- 11727182 TI - [Publications in scientific journals]. PMID- 11727183 TI - [What kind of surgical radicality is justified in the treatment of esophageal and cardial carcinoma?]. AB - In the period from January 1, 1973 to November 30, 1994, a total number of 1 856 patients had been admitted to our Department of Surgery because of cancer of the esophagus and esophago-gastric junction. We divided our activities into two study periods. In the first study period from 1973 to 1984 only so called "conventional operations" were performed. Since 1985 new oncological aspects were introduced into our operative tactics:1. the subtotal esophagectomy combined with the two field lymphadenectomy,2. the total gastrectomy with extended lymph node dissection. The analysis of our results with respect to the survival parameters (TNM staging, histological type, grade of differentiation, gross pathology) showed that the best chances were obtained by curative resection and lymphadenectomy in tumours of low-grade biologic malignancy. In esophageal cancers the former 6 % 5-year cumulative survival rate increased to 26 %, and in cardia tumours from 9 to 27 %, because of enhancement of radicality and extension of lymphadenectomy. PMID- 11727184 TI - [Twenty-year experience with multimodal treatment of gastric cancer patients in Poland]. AB - BACKGROUND: Poland is among the countries with high morbidity and mortality rates for gastric carcinoma (GC). Differences of GC biology depending on the geographical regions were assumed. In the literature, there are no reports from detailed clinicopathologic studies carried out in large series of GC patients in Poland. METHODS: Based on the prospectively collected data of 3 696 GC patients treated surgically between 1977-1999, potentially significant prognostic factors were analyzed to assess their prognostic value, and their time related distribution during the over 20-year period of the study. RESULTS: The mean age of the patients was 59.0 (SD 11.6) years and ranged from 20 to 93 years. The male to-female ratio was 2.3. Overall cumulative 5-year survival was 0.28 (for early GC 0.85) and significantly increased over the period of study. Among factors analyzed the depth of invasion, lymph nodes status, tumor size, age, UICC-R classification and ratio of involved to removed lymph nodes are of statistically significant and most important prognostic value. In the over 20-year period of study the rate of diffuse type carcinoma according to Lauren increased, but the tumor locations did not change significantly. The rate of early GC did not change throughout the period of study but number of most advanced cases decreased. Chemo and chemoimmunotherapy improved outcomes in some groups of patients. CONCLUSIONS: The basic clinicopathologic profile of Polish patients is similar to that reported in Western Europe and Japan, except for significantly higher early GC occurrence in Japan. According to the obtained results the prognosis of GC depends clearly upon the stage of the disease at the time of surgery. The improvement of outcomes during over 20 years of GC study in Poland was due to introducing better diagnosis and routine multimodal treatment. PMID- 11727185 TI - [Upper gastrointestinal bleeding - Review of our ten years results]. AB - Acute upper gastrointestinal bleeding (UGIB) is a common reason for emergency hospitalisation. Early upper gastrointestinal endoscopy is the corner stone of management; the alternative option for achieving primary hemostasis is emergency surgery. The aim of this study was to analyse the frequency of UGIB in the last 10 years and to present our surgical results. We observed 5 955 bleeding patients (68.4 % male and 31.6 % female) with a mean age of 57.7 inverted question mark 15.8 years. The most frequent causes of bleeding were gastric and duodenal ulcers (61 %) followed by gastroduodenal erosions (15.4 %) and varicous veins (5.7 %). Indications for emergency surgery were massive UGIB or rebleeding after active endoscopic treatment. On operation, gastric and duodenal ulcers were responsible for massive UGIB in 86.4 % cases. Overall hospital mortality rate during 10 years was 13.2 % and depended on age and concomitant diseases. In total 5.9 % of operated patients were rebleeding. Those with rebleeding underwent a second operation and showed a statistically higher mortality rate (35.7 % vs 11.8 %) compared to those surgical patients without rebleeding; p < 0.001. The mortality rate after vagotomy and pyloroplasty was 13.2 % (14/106) and did not differ significantly from that after gastric resection (15.2 %; 9/59). PMID- 11727186 TI - [Pelvic exenteration in the treatment of locally advanced tumors of surgical, gynaecological and urological origin]. AB - Pelvic exenteration is a radical extirpative procedure designed to treat pelvic malignancy that has invaded more than one of the hollow organs of the gastrointestinal or genitourinary tracts. The operation comprises several radical procedures: resection of the rectum, hysterectomy (with adnexectomy) with partial or complete vaginectomy, pelvic lymphadenectomy and total (or partial) cystectomy with diversion of the urinary tract. The retrospective analysis of our results in 27 patients treated in the years 1995 to 2000 by means of pelvic exenteration showed a 7.4 % hospital mortality and 59.3 % morbidity rate. The recurrence and survival rates were adequate. This surprisingly favourable outcome in our patients should encourage Czech surgeons to perform this difficult procedures more often. PMID- 11727188 TI - [A new method for inguinal hernia repair]. AB - We report 60 operations for groin hernia which were performed with Artschvadze's new method. 45 operations were carried out under local anaesthesia, 15 under general anaesthesia. 46 primary and 14 recurrent hernias were included in the study. During a follow-up of 6-30 months no recurrences were seen. PMID- 11727187 TI - [Blood purification methods for treatment of organ failure in patients with severe pancreatitis]. AB - BACKGROUND: Recent years have shown a considerable progress in the management of severe pancreatitis (SP); however, the role of extracorporeal blood purification in the treatment of progressive multiple organ dysfunction syndrome (MODS) is not well assessed. THE AIM OF THIS STUDY: The retrospective assessment of the MODS treatment results after application of extracorporeal blood purification methods in SP patients. METHODS: 50 consecutive patients with acute pancreatitis were included in the study. Atlanta classification system was applied for stratification of patients with SP. MODS was defined according to the recommendations of the Consensus Conference of American College of Chest Physicians/Society of Critical Care Medicine in 1991, and MODS score was calculated. Involvement of the organ systems, ICU, hospital stays and main outcomes were analyzed. RESULTS: Totally, 45 patients met SP criteria. Necrotizing pancreatitis was found in 35 patients. In 19 cases the clinical course was complicated with peritonitis, in two with jaundice. Infection was present in four patients. MODS were observed in 34 patients, with average of 3.3 organ involvement. In total, 21 patients underwent surgery. Hemodialysis was necessary in 5 of the 21 patients with necrotizing pancreatitis. Hemofiltration was applied in 5, hemadsorption in 28 and plasmapheresis in 23 patients. The overall mortality in association with hemodialysis and hemofiltration reached 20 % in necrotizing pancreatitis patients. Hemadsorption was associated with 3.8 % mortality, and plasmapheresis with 25 % mortality rate in all SP patients. CONCLUSION: Combined derangement of the renal and hepatic function is highly associated with unfavorable outcome in patients with severe pancreatitis. Timely application of the extracorporeal blood purification methods may revert progression of MODS and can be successfully combined with surgery if it is needed in this category of patients. PMID- 11727189 TI - [Long-term results of the reduction mammaplasty with Revers T-technique]. AB - Functional and aesthetic aspects make the reduction mammaplasty a favoured surgical procedure. The vertical technique, as well as the technique with central pedicle have clearly gained in importance next to the traditional T-cut technique. We provided questionnaires to a random sample of 110 patients who had been treated according to the method of Strombeck-Weiner. The results were evaluated and compared with the literature. Our results show that the functional complaints were largely eliminated by the surgery and that the patients experienced a significant improvement in their psychological well being. The applied method produces a satisfactory shape of the breast with programmed ptosis, and to large extent retains the nipple-areola sensitivity. One shortcoming of this procedure is the scar formation which 20 % of the patients found bothering. However, 94 % of the patients would choose the procedure again. We conclude that the reduction mammaplasty with reverse T-technique according to Strombeck-Weiner continues to be of value for patients with a desired weight reduction of more than 500-600 g and patients over the age of fourty. PMID- 11727190 TI - [Nonpalpable carcinoma of the breast - diagnosed by vacuum core breast biopsy and surgical management]. AB - The wire-localized extirpation is the "gold standard" for the examination of nonpalpable lesions suspicious of malignancy. Less invasive techniques were introduced in the last years, offering also a high diagnostic reliability, e. g. stereotactic core needle biopsy, the "advanced breast biopsy" and the vacuum core biopsy. Based on an analysis of 40 vacuum core breast biopsies and the following interventions in the case of carcinoma recommendations for the management of the nonpalpable breast carcinoma diagnosed by vacuum core biopsy should be developed. In 12 patients (33 %) carcinomas were found necessitating further operations. These were 92 % pTis or pT1pN0M0-carcinomas and only in one case an occult pT2pN1M0-carcinoma. We recommend a short interval between core biopsy and operation, a preoperative localization of the clips e. g. the residual microcalcification, and the controlled placement of the hooked wire that should also be performed at the Mammotome(R) using the same way to the tumor. Furthermore it is necessary to excise the core biopsy localization channel en bloc together with a wide tumour excision. An intraoperative histological examination of the specimen should be performed to confirm tumour-free excision borders. For this, the position of specimen should be marked by a thread and a specimen radiography should be made for the orientation of the pathologist and for documentation. A long-term follow-up of these patients under study conditions should be considered. Patients with benign diagnosis, not undergoing general anesthesia and operation with the consequences for later radiological evaluation, mostly profit from vacuum core breast biopsy. For patients with carcinoma the costs of the perioperative management increase. This should have consequences for the quality assurance of this method. PMID- 11727191 TI - [Nonpalpable carcinoma of the breast - diagnosed by vacuum core breast biopsy and surgical management. Invited commentary]. PMID- 11727192 TI - [Perioperative antimicrobial prophylaxis for colonic surgery: Present status in Germany. A prospective multicenter study with and without Metronidazole]. AB - INTRODUCTION: The objective of a multicentric observational study, that was performed in Germany between 1(st) September 1996 and 30(th) September 1997, was to assess postoperative infections as a function of risk factors and antibiotic prophylaxis under everyday clinical conditions. 2 481 patients from 114 centres who received infection prophylaxis prior to elective colonic resection were included. In the descriptive analysis of the study it was noted that 36.1 % of the patients had received no prophylaxis with metronidazole despite the fact that the study protocol recommended the use of this drug in preoperative antibiotic combinations. The present analysis therefore considers the influence of metronidazole on the postoperative infection rate. METHODS: In order to exclude any bias due to intergroup differences in risk profile, the groups with and without metronidazole were subjected to a matched-pair analysis. Matching parameters were: duration of operation, blood loss, age, diabetes mellitus, hepatic, renal, or chronic airways disease, immunosuppressive therapy, and rectal resection. This led to the formation of 800 pairs that were matched with respect to these parameters. The 800 pairs were then stratified into the following treatment groups: Group 1 a and b: long-acting cephalosporine (ceftriaxone) with or without metronidazole (n = 2 x 491); Group 2 a and b: short-acting cephalosporines with or without metronidazole (n = 2 x 133); Group 3 a and b: broad-spectrum penicillines with or without metronidazole (n = 2 x 176). RESULTS: In all three treatment groups combination therapy with metronidazole was found to be significantly superior. Postoperative infection rates were 9.4 % and 18.7 % (p = 0.000) respectively in Group 1 a and b, 12.0 % and 25.6 % (p = 0.008) respectively in Group 2 a and b, and 19.9 % and 29.0 % (p = 0.009) respectively in Group 3 a and b. CONCLUSION: Preoperative administration of metronidazole in addition to an effective beta-lactam antibiotic is strongly advised in elective colonic surgery, as absence of antibiotic cover against anaerobic colonic flora leads to a significantly higher postoperative infection rate. PMID- 11727193 TI - [Management of hemorrhage and perforation following endoscopic sphincterotomy]. AB - Bleeding and perforation are rare but dangerous complications of diagnostic and therapeutic ERCP and endoscopic sphincterotomy (EST). To evaluate the clinical outcome of patients treated for complicated EST in our surgical department, data were collected prospectively between 1/1995 and 3/2000. A total of 9 patients were admitted to our department, 7 women and 2 men, average age 60 (range 41-72) years. 5 patients were treated for severe hemorrhage, all of them underwent laparotomy following duodenotomy and oversewing of the sphincterotomy site. In average 10 hours (range 4-20) after endoscopy. One of these patients died due to multiple organ failure. In 4 additional patients a retroperitoneal perforation was discovered, 2 of these patients underwent laparotomy and drainage of the retroperitoneal cavity. The other two patients were sufficiently treated by percutaneous drainage. One of the patients, who underwent surgical drainage for retroperitoneal perforation died. In conclusion for severe hemorrhage duodenotomy and oversewing of the bleeding site is recommended in combination with common bile duct T-drain or other draining procedures. The treatment for perforation should depend on the clinical finding. A laparotomy seems not always to be necessary. PMID- 11727194 TI - [Ischaemic stump necrosis after subtotal gastric resection - a rare complication]. AB - The development of gastric stump necrosis after subtotal gastrectomy with Roux-en Y loop reconstruction and splenectomy in a patient with gastric cancer is described. The patient's life could be saved by re-operations on the 12th and 50th postoperative day. Due to the patient's poor general condition and local peritonitis the gastric stump was removed and a catheter-jejunostomy was prepared by using the Roux-en-Y loop. The aboral end of the oesophagus was closed. To relieve the oesophageal stump a cervical mucoso-oesophagostoma was carried out. As the patient's general condition improved the reconstruction of the alimentary tract was performed by preparing a new Roux-en-Y loop oesophago-jejunostomy and by closing the mucoso-oesophagostoma.9 years after the operation the patient died of pulmonary metastases. The reasons for the delayed re-operation and the advantages of the cervical mucoso-oesophagostoma are explained. PMID- 11727195 TI - [Intraabdominal cystic lesions - the differentation of mesenteric cysts and cystic lymphangioma]. AB - Mesenteric cysts and cystic lymphangioma are rare intraabdominal lesions. In our clinic, during one year, two young women with these intraabdominal cysts underwent operation because of unspecific abdominal pain. Despite preoperative imaging the final diagnosis of a cystic lymphangioma and a mesenteric chylous cyst was made not before surgery. The postoperative course was uneventful. Problems of diagnostic, differential diagnosis and therapy are presented and differences in etiology, histology and location are discussed. PMID- 11727196 TI - [Duodenosigmoidal fistula in a patient with Crohn's disease]. AB - The development of enteral fistulas - internal and external - is common in Crohn's disease with a frequency of about 45 % in large series of patients. Most internal fistulas arise in the small bowel, but internal large bowel fistulas occur in about one fourth of all patients with fistulas in Crohn's disease. Colonic gastroduodenal fistulas do occur, but are very rare. Of the 35 cases of colonic-duodenal fistulas that we found in the literature, most have been described to involve either a previous ileocolostomy site or the ascending or transverse colon. There are only three cases displaying sigmoidoduodenal fistulas. To these we add a fourth case with this report. A 42-year-old woman presented with upper abdominal pain, loss of appetite and regurgitations with a foul smell as well as abdominal gas complaints. A duodenosigmoidal fistula was diagnosed by local application of contrast and the fistula tract was excised in typical fashion by resection of the colonic fistula and primary closure of the duodenum. The patient experienced a good recovery with relief of symptoms. PMID- 11727197 TI - [Richard von Volkmann and the Surgical School of Halle in 1867 to 1889]. AB - Richard von Volkmann (1830-1889) was appointed senior consultant of the University Surgical Hospital in Halle at the young age of 36. On returning home from the war of 1870/1871, he saw traumatic fever rampant in his hospital. He became a rigorous advocate of Lister's method of antisepsis. There were a large number of very serious accidents in the up-coming industrial town of Halle. Acute and chronic inflammation of the joint led to severe disfigurements. By invention of large reconstructive operations von Volkmann was able to obtain impressive results. He was the founder of traumatology and orthopaedic surgery. Tuberculosis was regarded at that time as the most common cause of death. By means of surgery certain forms of the illness could be cured. Tuberculous coxitis and gonarthritis were the most common indications for orthopaedic surgery. Von Volkmann used his distraction method for tuberculous spondylitis.Many terms used in medical practice derive from Richard von Volkmann. He described Volkmann's triangle, ischaemic muscle contractures and resorption fever. He developed a whole range of new methods of surgical procedures and the concept of wound drainage. His name is associated with Volkmann's splint, Volkmann's "sharp spoon" and several other inventions. Richard von Volkmann was one of the founders of the Deutsche Gesellschaft fur Chirurgie (German Surgical Society). His famous Traumereien an franzosischen Kaminen ("Reveries at French Firesides"), an anthology of fairy tales for his children, played an important role in his life's work. PMID- 11727198 TI - Relationship inference from trios of individuals, in the presence of typing error. AB - Misspecification of relationships and of genotype data can cause problems in linkage analyses based on genome-scan data. Previous reports have focused on pairwise relationships and a simple error model. This article considers the increased information available from the joint analysis of trios of individuals, integrating this analysis with an error model that allows for the most common genotyping errors. Given observed marker phenotypes in a genome scan, computational methods are outlined both for likelihoods of relationships and for the posterior probabilities of underlying genotypes. The methods are applied to examples from two real data sets: one has been previously well analyzed, and, hence, Mendelian inconsistencies have been removed; the other typifies the pedigree and genotype errors encountered in the initial analyses of a study. It is demonstrated that the coupling of relationship inference and error detection is quite effective, that the error model is computationally practical, and that data on a third relative can often clarify relationships. PMID- 11727199 TI - C-->U editing of neurofibromatosis 1 mRNA occurs in tumors that express both the type II transcript and apobec-1, the catalytic subunit of the apolipoprotein B mRNA-editing enzyme. AB - C-->U RNA editing of neurofibromatosis 1 (NF1) mRNA changes an arginine (CGA) to a UGA translational stop codon, predicted to result in translational termination of the edited mRNA. Previous studies demonstrated varying degrees of C-->U RNA editing in peripheral nerve-sheath tumor samples (PNSTs) from patients with NF1, but the basis for this heterogeneity was unexplained. In addition, the role, if any, of apobec-1, the catalytic deaminase that mediates C-->U editing of mammalian apolipoprotein B (apoB) RNA, was unresolved. We have examined these questions in PNSTs from patients with NF1 and demonstrate that a subset (8/34) manifest C-->U editing of RNA. Two distinguishing characteristics were found in the PNSTs that demonstrated editing of NF1 RNA. First, these tumors express apobec-1 mRNA, the first demonstration, in humans, of its expression beyond the luminal gastrointestinal tract. Second, PNSTs with C-->U editing of RNA manifest increased proportions of an alternatively spliced exon, 23A, downstream of the edited base. C-->U editing of RNA in these PNSTs was observed preferentially in transcripts containing exon 23A. These findings were complemented by in vitro studies using synthetic RNA templates incubated in the presence of recombinant apobec-1, which again confirmed preferential editing of transcripts containing exon 23A. Finally, adenovirus-mediated transfection of HepG2 cells revealed induction of editing of apoB RNA, along with preferential editing of NF1 transcripts containing exon 23A. Taken together, the data support the hypothesis that C-->U RNA editing of the NF1 transcript occurs both in a subset of PNSTs and in an alternatively spliced form containing a downstream exon, presumably an optimal configuration for enzymatic deamination by apobec-1. PMID- 11727201 TI - The gene mutated in variant late-infantile neuronal ceroid lipofuscinosis (CLN6) and in nclf mutant mice encodes a novel predicted transmembrane protein. AB - The neuronal ceroid lipofuscinoses (NCLs) are a group of autosomal recessive neurodegenerative diseases characterized by the accumulation of autofluorescent lipopigment in various tissues and by progressive cell death in the brain and retina. The gene for variant late-infantile NCL (vLINCL), CLN6, was previously mapped to chromosome 15q21-23 and is predicted to be orthologous to the genes underlying NCL in nclf mice and in South Hampshire and Merino sheep. The gene underlying this disease has been identified with six different mutations found in affected patients and with a 1-bp insertion in the orthologous Cln6 gene in the nclf mouse. CLN6 encodes a novel 311-amino acid protein with seven predicted transmembrane domains, is conserved across vertebrates and has no homologies with proteins of known function. One vLINCL mutation, affecting a conserved amino acid residue within the predicted third hydrophilic loop of the protein, has been identified, suggesting that this domain may play an important functional role. PMID- 11727202 TI - [Imbalance of plasma amino acids in patients with autism and subjects with attention deficit/hyperactivity disorder]. AB - INTRODUCTION: Plasma and brain amino acids are influenced by dietary intake. Alterations of plasma amino acid concentrations have been reported in neuropsychiatric disorders. OBJECTIVE: To analyse the plasma amino acid values in subject diagnosed with autism, with attention deficit/hyperactivity disorder (ADHD), and healthy subjects as controls. PATIENTS AND METHODS: Forty subjects affected by autism, 11 with ADHD and 41 healthy subjects (age range 3 18 years old) were included in this study. Peripheral venous blood was obtained in fasting condition, collected in EDTA tubes and centrifuged. Plasma was de proteinised with sulfosalicylic acid. Amino acids were analysed by ion exchange liquid chromatography with an LKB amino acid analyser with sodium citrate elution system and ninhydrin reaction. Results were expressed as mmol/L. RESULTS AND CONCLUSIONS: In both disorders a diminution of phenylalanine and glutamine plasma concentrations was observed beside an increase of glycine. Lysine appeared increased only in autistic subjects. These alterations produce an imbalance with the rest of plasma amino acids competing at the brain blood barrier by the same transport system thus causing alterations in the metabolism and/or transport of amino acids to the brain, altering CNS functions. The phenylalanine decreasing, beside glycine increasing appear to support the hypothesis of a disorder in the inhibitory neurotransmission system, especially in ADHD. The diminution of phenylalanine and the increasing of lysine in autism are suggestive that these two amino acids are metabolically related. PMID- 11727200 TI - Retinal dystrophy due to paternal isodisomy for chromosome 1 or chromosome 2, with homoallelism for mutations in RPE65 or MERTK, respectively. AB - Uniparental disomy (UPD) is a rare condition in which a diploid offspring carries a chromosomal pair from a single parent. We now report the first two cases of UPD resulting in retinal degeneration. We identified an apparently homozygous loss-of function mutation of RPE65 (1p31) in one retinal dystrophy patient and an apparently homozygous loss-of-function mutation of MERTK (2q14.1) in a second retinal dystrophy patient. In both families, the gene defect was present in the patient's heterozygous father but not in the patient's mother. Analysis of haplotypes in each nuclear kindred, by use of DNA polymorphisms distributed along both chromosomal arms, indicated the absence of the maternal allele for all informative markers tested on chromosome 1 in the first patient and on chromosome 2 in the second patient. Our results suggest that retinal degeneration in these individuals is due to apparently complete paternal isodisomy involving reduction to homoallelism for RPE65 or MERTK loss-of-function alleles. Our findings provide evidence for the first time, in the case of chromosome 2, and confirm previous observations, in the case of chromosome 1, that there are no paternally imprinted genes on chromosomes 1 and 2 that have a major effect on phenotype. PMID- 11727203 TI - [Efficacy and tolerance of gabapentin in dystonia and hemifacial spasm: a pilot study]. AB - INTRODUCTION: The pharmacological management of dystonia is suboptimal. Focal dystonia and hemifacial spasm can be successfully managed with botulinum toxin injections. The physiopathology of dystonia remains a mystery. The gabaergic transmission may be involved in the origin of dystonia. Gabapentin is a gabaergic agent and therefore it can be useful in dystonia. PATIENTS AND METHODS: 28 patients with different forms of dystonia and hemifacial spasm were included in this open study. They received gabapentin (mean dose 1885 mg/d). RESULTS: Gabapentin was not efficacious but a subgroup of 6 patients (3 with hemifacial spasm, 2 with posttraumatic dystonia and 1 with spasmodic torticollis) significantly improved. Gabapentin was well tolerated. Somnolence was the most commonly reported side effect. CONCLUSION: Gabapentin may be useful in some patients with dystonia and hemifacial spasm. Double blind studies are clearly needed. PMID- 11727204 TI - [Profile of epilepsy in a neurology clinic in Costa Rica]. AB - INTRODUCTION: Epilepsy is a worldwide medical problem it affects nearly 1% of the population. In Costa Rica it s no different. Although it is an important problem we don t have any statistical information concerning only epilepsy. PATIENTS AND METHODS: We took on the task of studying the cases of epilepsy in a Neurology clinic in a third level hospital, from January to August 2000. The clinical records of 127 patients with epilepsy were studied. We excluded patients with febrile seizures and those with only one isolated seizure not related to any neurological injury. RESULTS: There was a slight predominance of men (55.11%) over women (44,8%). A great majority of the patients did not have family history of epilepsy, and the average age of on set was 20.062 years. Idiopathic epilepsy was predominant with a 48,3%, 26,77% were cryptogenic and 25,19% symptomatic. Of this, the most frequent etiology was sclerosis and atrophy of the temporal lobe, followed by cerebrovascular disease. As to image studies 96,06% had CT, MRI or both; and 94,48% had an EEG. Of all the patients a great majority are receiving treatment with one antiepileptic drug (81,88%). Valproic acid and carbamazepine are the most commonly used. 51,98% of patients have not had a seizure in at least 8 months. CONCLUSIONS: It is clear, that our health system provides most of the basic needs required to control most of the epileptic patients. However there are important challenges in the present and pear future, such as developing video encephalography units for brain mapping and supporting new epilepsy surgery programs. It is our mission also to induce public health authorities to realize the impact of Epilepsy in social affairs and economics. PMID- 11727205 TI - [A comparative study of intravenous immunoglobulin and plasmapheresis preoperatively in myasthenia]. AB - INTRODUCTION: Thymectomy is a well established procedure in myasthenia gravis (MG). To reduce its morbidity, treatments which are effective in the short term, such as plasmapheresis and intravenous immunoglobulin (IGI) have been used. OBJECTIVE: To evaluate the efficacy and tolerability of the IGI, compared with plasmapheresis, in the preparation of myasthenic patients before thymectomy. PATIENTS AND METHODS: We compared a group of 33 prospective myasthenic patients treated with IGI with 38 clinical histories taken as controls treated by plasmapheresis during the peri operative period of thymectomy. RESULTS: The patients treated with IGI were in the intensive care unit and neurology ward for less time. The endotracheal tube was also removed sooner. However, these differences were not significant. The commonest complications of IGI were fever, shivering and phlebitis. The most frequent adverse reaction to plasmapheresis were cutaneous eruptions. One patient developed Hepatitis C after plasmapheresis. CONCLUSION: IGI is comparable in efficacy to plasmapheresis in the peri operative period of MG, but has a better profile of adverse reactions. PMID- 11727206 TI - [Changes in nerve growth factor levels related to age and neurotrophic treatment in non-human primate]. AB - INTRODUCTION: To examine the amounts and role of growth factors in different tissues and corporal fluid, new sensitive techniques have to be developed. A major problem is that the normal concentration of trophic substances, such as nerve growth factor (NGF), in central and peripheral nervous system and in fluids is very low (ng pg/ml). A valuable method of research is the sensitive two site enzyme immunoassay using the monoclonal antibody 27/21 to mouse NGF. Materials and methods. The present work applied this enzyme immunoassay to examine the NGF levels in normal non human primate sera (n= 94) and applied this assay to study of NGF levels in two non human primate receiving NGF infusion: one young and one aged. Two groups of non human primate sera were studied one young adult (n= 69) and one aged (n= 25). The serum samples NGF treated non human primate were taken before the infusion and at the 1st week and 1st, 3rd, 6th and 12th month after infusion. RESULTS: To further test the specificity of conjugate binding, dilutions of the non human primate sera were preincubated with an excess of monoclonal NGF antibody 27/21 in solution. With this strategy it was possible to completely block the signal obtained using the enzyme immunoassay. We found very low levels of NGF in aged monkeys (0.054 ng/ml) when compared with young adult group (0.152 ng/ml) (p> 0.01). The NGF levels in aged non human primate treatment with NGF was very low before (0.50 ng/ml) and during NGF treatment evolution time, whereas at the the 12th month showed an increase in NGF levels (0.180 ng/ml). We found normal values of NGF in the young monkey before and during the first year after NGF infusion. CONCLUSIONS: Using the enzyme immunoassay described it is possible to know the serum concentration of NGF immunoreactive in non human primate and this assay is able to detect peripheral changes in NGF levels after intracerebral infusion of NGF. PMID- 11727207 TI - [The advantages of a programme for training carers of persons with dementia: a pilot study by specialised nurses]. AB - INTRODUCTION: The care of patients with dementia usually has to be undertaken by persons with no formal training. However, the chances of treatment being effective depends on them. Specialist non hospital nurses, who are accessible and close to these patients, are in a privileged position to develop programmes to train carers. In our country no studies have been done to evaluate the efficacy of such programmes. OBJECTIVES: To evaluate the efficacy of a programme for training carers of patients with irreversible dementia. The programme was designed and carried out by specialist nurses, based on a neurology clinic outside the hospital. PATIENTS AND METHODS: Ten carers who came consecutively to the clinic were invited to participate in the programme. Before and after the programme we evaluated cognitive, functional behaviour and affective aspects of the patient and/or carer. RESULTS: As far as the patient was concerned no changes were found in either cognitive or functional aspects. Although the behaviour problems were reduced, they caused less stress to the carer after the training sessions. At the same time the affectivity of the patients improved. CONCLUSIONS: It is possible to carry out effective training programmes for carers of demented patients, with relatively little expense. Affectivity, both of the patient and the carer is the aspect most improved by this training. Large scale trials are necessary to show improvement of other aspects, and to compare different types of training programmes. PMID- 11727208 TI - [Influence of estradiol on pyroglutamyl aminopeptidase activity in the frontal cortex of ovariectomized mice]. AB - INTRODUCTION: Pyroglutamyl aminopeptidase (pGluAP) is an omega peptidase widely distributed in fluid and tissues which hydrolyses biological active peptides including thyrotropin releasing hormone (TRH). OBJECTIVES. The aim of present work is to study the influence of estradiol on soluble and membrane bound pGluAP activity in the frontal cortex of female mice. MATERIAL AND METHODS: Soluble and membrane bound pGluAP activities in frontal cortex of ovariectomized mice and ovariectomized mice injected with different doses of estradiol were measured using espectrophotometric assays. RESULTS: Soluble pGluAP activity in frontal cortex did not change after ovariectomy or after the administration of the different doses of estradiol. However, membrane bound pGluAP activity showed a significant increase after ovariectomy. After the administration of the lower dose of estradiol, membrane bond pGluAP activity returned to the same levels detected before the ovariectomy. CONCLUSION: Estradiol modifies membrane bound pGluAP activity which is the principal enzyme involved in the hydrolysis of TRH. Therefore, misregulation of estradiol levels may produce modifications in the neuromodulatory functions of TRH. PMID- 11727209 TI - [Dysautonomic features in patients with Cuban type 2 spinocerebellar ataxia]. AB - INTRODUCTION: Dysautonomic features are the clinical signs and symptoms derived from anomalous functioning of the sympathetic or parasympathetic nervous systems in either the peripheral or central parts. OBJECTIVE: To identify the clinical features related to dysautonomia in a patient with spinocerebellar ataxia with severe functional disability. PATIENTS AND METHODS: We studied a series of cases, including 21 patients with severe disability (confirmed to their bed or wheelchair, unable to walk and totally or partly dependent on other people for essential everyday activities). The patients and their families were closely questioned, and full clinical examination included a test for orthostasia. RESULTS: All patients had some signs of peripheral dysautonomia: all had vasomotor disorders (orthostasia, distal pallor and coldness, Raynaud s phenomenon, etc.), 95.2% (constipation, urinary and rectal incontinence, polachuria, palpitations, tachycardia at rest, etc.), exocrine gland disorders in 71.4% (increased lachrymation, reduced sweating, increased or reduced salivation), 87.5% peripheral tissue nutrition disorders. Similarly, in all patients studied there was evidence of central dysautonomic disorder, with a syndrome of cachexia with bulimia, sleep disorders together with dysregulation of thirst and body temperature. CONCLUSIONS: In patients with type 2 hereditary spinocerebellar ataxia with severe disability there was involvement of the peripheral and central nervous system regulating autonomic function. PMID- 11727210 TI - [Parkinsonism probably induced by manganese]. AB - INTRODUCTION: In all cases of young persons with clinical Parkinson s disease it should be suspected that it is secondary to some primary disorder. Therefore a battery of diagnostic tests should be done before classification as idiopathic Parkinson s disease. CLINICAL CASE: A 31 year old woman whose only previous illness had been Graves disease. She complained of difficulty with movements of her right arm and leg for some months (she had problems with walking and with rapid, repeated movements of her right hand). She also complained of tremor of her right limbs at rest. She denied taking drugs, having dysphagia, dysarthria, visual changes or sphincter disorders. Neurological examination showed her to have monotonous speech, slight facial hypomimia, slight reduction in spontaneous blinking, walking with less swing of her right arm; postural tremor of both arms, worse on the right; bradykinesia (2/4) of both right limbs and rigidity (1/4), axial and of the right limbs. The results of all the investigations done to rule out secondary Parkinsonism were normal, except for the plasma manganese level which was raised, although it returned to normal when the probable source of exposure to this metal was removed. However, the alterations of movement only disappeared after treatment with levodopa was started. CONCLUSION: In cases of Parkinsonism in young adults secondary causes should always be rules out, such as exposure to certain metals. PMID- 11727211 TI - [Simultaneous cephalic migration of a ventriculoperitoneal shunt into the intraventricular and subdural spaces. A case report]. AB - INTRODUCTION: Hydrocephaly is defined as a hemodynamic disorder in which the production, circulation or reabsorption of cerebrospinal fluid is involved. Migration, in a cephalic direction, into the subdural and intraventricular spaces of the proximal ends (shunt malfunction), is very unusual and perhaps not yet reported. It probably involves movement of the craniospinal region of the patient which leads to a follow on effect due to an underlying disorder. CLINICAL CASE: We report the case of a polytraumatized baby (non hospital birth, subarachnoid hemorrhage, porencephaly and subsequent hydrocephaly), who had had multiple shunts inserted. On her most recent admission the shunt was not working properly, as confirmed on plain radiological examination. This showed migration of the catheter, in cephalic direction, into the intraventricular and subdural space. CONCLUSIONS: In order that such migration could occur, conditioning factors would be necessary: such as detachment of the shunt at the distal end (technical fault), underlying disease (porencephaly), dynamic factors causing expulsion (abdominal peristaltic movements) dynamic translocation factor (neck movements), dynamic attraction factor (increased CSF reabsorption) and unishunt catheter (offering no resistance to passage through the trepanation orifice). PMID- 11727213 TI - [Infantile neuroaxonal dystrophy. A report of two new cases and a review of the literature published over the past ten years]. AB - INTRODUCTION: The neuroaxonal dystrophies make up a group of neurodegenerative disorders of unknown origin, which are characterized by all showing axonal lesions. The infantile form, or Seitelberger s disease, is one of the forms of earliest onset and rapid progression. The clinical, neurophysiological and pathological criteria described by Aicardi and Castelein in 1979 are still valid. However, we should emphasise the great usefulness of cerebral MR scanning in making an early diagnosis of this condition. CLINICAL CASES: We report two brothers, sons of consanguineous parents, who fulfilled the above clinical criteria. Their illness presented before the age of three years, with arrested psychomotor development followed by regression, an initial hypotonia syndrome which progressed to spastic tetraplegia, optic atrophy and progressive deafness, blindness and dementia. Neurophysiological findings were of central conduction disorders, and chronic denervation was shown on EMG. On EEG there were high frequency, high voltage rhythms. MR scanning showed the cerebral cortex to become atrophied and hyperintense at an early stage. On biopsy of the sural nerve and of skin there was spheroid swelling of the axons with tubulous vesicular material seen in myelinated and nonmyelinated axons. CONCLUSIONS: We reviewed the literature published over the past ten years (1990 2000). From this we conclude that on the initial clinical and neurophysiological criteria of Aicardi and with the aid of current neuroimaging techniques, the diagnosis may be suspected sufficiently early so as to permit genetic counselling. This would help to avoid further, high risk pregnancies, even before the diagnosis had been confirmed by the biopsy findings. PMID- 11727212 TI - [Familial Pitt-Rogers-Danks: two new cases]. AB - INTRODUCTION AND CLINICAL CASES: The Pitt Rogers Danks syndrome is characterized by prenatal and postnatal retardation of growth, mental retardation, microcephaly, convulsions and a peculiar facies. It is believed to represent a clinical variant of the Wolf Hirschhorn syndrome, since there is a deletion in the 4p16.3 region in both syndromes. We report two cases in the same family caused by maternal mal segregation of a 4:8 balanced translocation. We describe the clinical characteristics, investigations done and a review of the literature. PMID- 11727214 TI - [The cerebral cortex modulates the cutaneous transmission through the dorsal column nuclei]. AB - INTRODUCTION: The mechanisms used by the cerebral cortex to modulate the cutaneous information at prethalamic level have been scarcely studied. This article reviews experimental evidence leading to a better understanding of this issue at the level of the cuneate nucleus (Burdach nucleus). DEVELOPMENT: The primary afferents and the corticocuneate fibers make synaptic contact with cuneothalamic neurons and with inhibitory interneurons in the middle cuneate nucleus. By stimulating the skin at different places while recording the cuneothalamic intracellular activity in anaesthetized animals with the cortex intact, with the cortex pharmacologically inactivated, or in absence of a cerebral cortex it was possible to ascertain the functional role of the corticocuneate fibers. The primary afferents activated by stimulating a particular zone of the skin induce monosynaptic excitation on a group of cuneothalamic cells at the same time at which inhibit, through intranuclear interneurons, neighboring cuneothalamic cells with unmatched receptive fields. Similarly, the corticocuneate cells receiving information from the stimulated skin further increase the excitation of the cuneothalamic neurons with matched receptive fields while inhibiting others with unmatched fields. The cortex exaggerates an excited center surrounded by an inhibited periphery thus increasing the tactile discrimination both spatially and temporally which is essential for exploratory and manipulative purposes. PMID- 11727215 TI - [Classification of the cerebrovascular diseases. Iberoamerican Cerebrovascular diseases Society]. AB - OBJECTIVES: In view of the several terminologies in countries belonging to the Iberoamerican Cerebrovascular diseases Society it seemed appropriate that the Executive Committee should propose a unified cerebrovascular diseases classification and nomenclature. DEVELOPMENT: Based on the classifications of strokes NINCDS, TOAST, Laussane Stroke Registry, OCSP and that of the cerebrovascular diseases group of the Sociedad Espanola de Neurologia, we devised a system for classification of cerebrovascular diseases with the objective of harmonizing the nomenclature and criteria of Iberomerican countries. We chose the word stroke as the most suitable for the definition of cerebrovascular diseases. These were then divided into symptomatic and non symptomatic. The symptoms were then divided into those of transient ischaemic accidents (TIA) and stroke, and then into ischaemic and haemorrhagic. We established the aetiological subtypes of focal cerebral ischaemia (TIA and ischaemic stroke) as: disease of the great vessels or atherothrombotic, cardio embolic, occlusive small vessel disease, of unusual cause or of unknown origin. Haemorrhagic stroke was divided into subarachnoid haemorrhage and cerebral haemorrhage, which was then subdivided according to the site. CONCLUSION: We consider that this nomenclature is simple and easy to use and in concordance with the international classifications, which permits the use of common terminology that is easily translated into English. PMID- 11727216 TI - [Microdialysis cerebral. Main application of this technique]. AB - INTRODUCTION: Microdialysis cerebral technique has been widely employed in order to study the neurotransmitter release. DEVELOPMENT: This technique present numerous advantages such as: allow to work with sample in vivo from animals freely moving, by means of microdialysis can be infused simultaneously different drugs in different points implanted probes in several coordinates; also allow carry out pharmacokinetics studies that show correlation with behavior patter as well as to study metabolic changes, which is not possible when this determination are carry out in tissue, in post mortem stadio. CONCLUSIONS: In the present work is carry out a review about the main results obtaining in the sensitization and abstinence to several drug study and approach to biochemical characteristics to Parkinson s disease (PD) by means of microdialysis technique. In relation to sensibilization studies, the temporal changes in different behavior pattern that modify after amphetamine administration have been studied. Microdialysis study allowed correlationate dopamine concentration with behavior pattern above mentioned. In relation with PD, dopamine concentration after systemic and central (intracerebral) administration of levodopa and another dopaminergic drugs have been studied in different nucleus of basal ganglia as well as its respectively behavior correlates. PMID- 11727217 TI - [The amigdaloid body: functional implications]. AB - INTRODUCTION: The amygdaloid body is a structure localized to the temporal lobe in mammals, formed by different nuclei and traditionally associated with the emotion system of the brain. Some investigators have suggested an alternative function for the amygdaloid body, considering it to be part of the system modulating memory. Much of the experimental data supports both functions. DEVELOPMENT: The relation of the amygdaloid body with the emotions has been based on the study of the conditioning of fear, by which an emotionally neutral stimulus can cause emotional reactions due to its temporal association with an adverse stimulus. It has been shown that the amygdaloid body is necessary for learning and expressing this conditioning, and is therefore involved in emotional learning. With regard to the relation of the amygdaloid body with the modulation of memory, one should point out the results which show that it is not always necessary for learning and memory, but is necessary for showing the modulating effects of different substances on memory. The amygdaloid body modulates the storage of memory in other regions of the brain, such as the caudate nucleus and the hippocampus. CONCLUSIONS: Current data suggests that the whole amygdaloid complex is not involved in the two functions. However, the lateral and central nuclei participate in emotional learning whilst the basolateral nucleus is especially involved in modulation of the memory when emotional activation occurs. PMID- 11727218 TI - [Atrophy of the amygdala complex and neuropsychiatric expression of Alzheimer s disease]. AB - INTRODUCTION: In Alzheimer s disease, together with cognitive changes (loss of memory, lack of orientation in time and space, difficulty in carrying things out, etc.) non cognitive disorders also occur, seriously disturbing the patient s behavioral and emotional balance. Depression, delirious ideas, hallucinations and behavior changes (habits regarding sex, feeding and movement) are amongst the commonest features of this disease from its earliest stages. Several studies have described how initially the histopathological changes selectively involve different structures of the medial temporal lobe ( entorinal cortex, hippocampal formation, amygdala) and gradually includes neocortical association areas. Thus, the amygdala complex, a structure related to processes of memory and emotional control, is severely affected in this disease from the initial stages. PATIENTS AND METHODS: Therefore, to find any possible relationship between involvement of the amygdala and the psychiatric expression of Alzheimer s type dementia, we did a radiological study, using magnetic resonance, together with a neuropsychological study of a group of 24 persons with Alzheimer s disease who had mild moderate deterioration. Analysis of the amygdala area and the scores on the subtest Alzheimer s Disease Assessment Scale Non cognitive (ADAS NC) were the variables chosen for measurement of the anatomical and psychiatric aspects of the subjects under investigation. RESULTS AND CONCLUSIONS: The results of our study show a close relation between involvement of the amygdala (atrophy) and the presence of neuropsychiatric changes in persons with Alzheimer s disease. PMID- 11727219 TI - [Cognitive deterioration after consumption of cannabis]. AB - OBJECTIVES: The use of cannabis in humans causes several malfunctions in the cognitive performance, but it remains unclear if these alterations are reversible after the abstinence. DEVELOPMENT: The author carries out a wide bibliographical review from the first studies including a large number of subjects, which results are not reliable due to methodological defects. Recent researches, using more strict criteria and modern techniques of neuroimaging, show a subtle and permanent fail in several cognitive functions. CONCLUSIONS: We consider that long term cannabis use (more than 15 years) would lead to a subtle and permanent impairment in cognitive performance, mainly in the capacity of retain new information (probably due to inability to filter tangential information). PMID- 11727220 TI - [Seven ipsative keys to understanding the poor 'Hispanic' student in the USA]. AB - INTRODUCTION: Examination of the special bilingual student (5% of the bilingual student population of the USA) permits us to use not only (1) the statistical reports of the population coefficients and percentages), (2) measurements of progress or performance (age in years or grades of progress at school) and (3) the neuropsychological dimensions of the brain (left and right, anterior and posterior, inferior and superior). DEVELOPMENT: It is possible to put into parenthesis the statistical norms, either national, local, regional or even Hispanic national, given the great disparity seen in the results. At the present time the Hispanic population is made up mainly of young persons, recent immigrants with limited education. So it is logical that there are three major sources of error for students who do an exam in a foreign language when they are already disadvantaged and handicapped. Alternatively, we may pay attention to ipsative comparisons (of the same subject regarding different aspects of his behavior), which are almost free of any norm but comparison between themselves. CONCLUSIONS: Here we list a series of comparisons connected with concrete and abstract conduct, asynchrony between knowledge and operations, hand work (unilateral and bilateral), linguistic and academic performance, in one language or the other. The comparisons observed help us to understand, in a new, relevant manner, the cognitive and academic function of the child who is handicapped, disadvantaged and bilingual . It also allows us to describe the basic dimensions of his cognitive and academic progress. PMID- 11727222 TI - [Meningioma in childhood: an unusual case]. PMID- 11727221 TI - [Familial amyloid polyneuropathy: two new cases of late onset, with many asymptomatic carriers in the family]. PMID- 11727223 TI - [Multifocal motor neuropathy and the Lewis Sumner syndrome. A case report]. PMID- 11727224 TI - [The cost of cerebral infarct in the acute phase]. PMID- 11727226 TI - [Visual extinction: a study by means of potentials evoked with movement]. AB - INTRODUCTION: The term extinction refers to the phenomenon in which a patient with a lesion of the central nervous system ignores one of two stimuli in conditions of simultaneous bilateral stimuli. Various studies observe this phenomenon as the expression of a deficit in the process of selective attention, but there are few studies with potentials related to events (PRE). OBJECTIVES: To find the possible electrical correlation of the extinction phenomenon and the stage of processing at which this occurs. PATIENTS AND METHODS: We studied a patient with a right parieto occipital lesion and a control person of the same race, age, sex and educational level. We carried out two experiments. The first of these was to determine the direction of movement of two surfaces with red and green spots situated on both sides of a central fixation point. In the second experiment, the same task was carried out, but the two surfaces were superimposed, in the same place. In both cases the first movement occurred in the surface which was being attended to and the second might be this surface or the other. The proportion of correct answers were recorded for both movements. The PRE showed an attention disorder which affected the spatial distribution of attention with conservation of the attention to objects. CONCLUSIONS: The suppression of the P1 N1 components are the electrical correlate of the phenomenon of extinction which should occur during early stages of processing. Further investigations are necessary in a larger number of cases. PMID- 11727227 TI - [Intracranial venous thrombosis: a study of 16 cases and a review of the literature]. AB - INTRODUCTION: The clinical picture and aetiology of intracranial venous thrombosis are highly variable. Early descriptions reported it as a rare disease with a poor prognosis but the advent of neuroimaging techniques, and a deeper knowledge of the clinical picture, have shown it to have a higher frequency and a better prognosis. OBJECTIVE: To report the clinical and neuroimaging findings in patients diagnosed as having intracranial venous thrombosis in our department and review the state of the literature. PATIENTS AND METHODS: We reviewed all discharge reports from patients admitted to the neurology department of the Juan Canalejo Hospital between 1975 and 2000. Of these, we reviewed the medical records of those patients diagnosed as having intracranial venous thrombosis in order to obtain data relating to the clinical manifestations, complementary tests, etiological and topographical diagnosis, treatment and outcome. RESULTS: Diagnosis of intracranial venous thrombosis was made in 16 patients. The most common symptom was headache. The superior sagittal was the most frequently affected sinus. In almost all patients CT results led to the suspicion, and in some cases the confirmation, of the diagnosis. The most frequently found aetiology was oral contraceptive consumption. Outcome was generally good both with anticoagulation and symptomatic treatments. CONCLUSIONS: The most important difference between the present study and earlier reports is in the frequency of the different aetiologies. Our findings provide further evidence that intracranial venous thrombosis is not an infrequent disease and that the prognosis is generally good. PMID- 11727228 TI - [Monitorization of the jugular oxygen saturation and cerebral ischaemia in the neurocritical patient]. AB - INTRODUCTION: Catheterization of the jugular bulb is of great value in the study of cerebral metabolism, since it permits diagnosis of episodes of hypoxia and cerebral ischaemia. OBJECTIVES: To determine the frequency of the appearance of episodes of desaturation and cerebral ischaemia, whether these were simultaneous and their relation to the mortality in an intensive care unit (ICU). PATIENTS AND METHODS: We studied 21 neurocritical patients, recording the following data: age, sex, diagnosis and state at the time of discharge from the ICU (alive or dead). In the group of patients in whom the jugular bulb had been catheterised we also recorded: the side cannulated, complications, SyO2, cerebral lactate oxygen index and modified cerebral lactate oxygen. RESULTS: In 57% of the patients we catheterised the jugular bulb and in 50% of them at least one episode of desaturation was recorded, and cerebral ischaemia in 42%. Only one patient had simultaneous desaturation and cerebral ischaemia. Of the six patients in whom desaturation was detected, 5 died in the ICU (83.3%). The mortality of patients with ischaemia was only 40%. 40% of the patients with head injuries were monitored and 50% of the non monitored patients had head injuries. CONCLUSIONS: Episodes of desaturation are frequent in neurocritical patients and are associated with increased mortality. Recordings of lactate are not useful at present in the diagnosis of cerebral ischaemia. PMID- 11727229 TI - [Neonatal EEG trace of burst suppression. Etiological and evolutionary factors]. AB - INTRODUCTION: The electroencephalographic (EEG) trace seen during the neonatal period which shows so called discharges of burst suppression, is caused by a severe disorder of cerebral electrogenesis occurring at this time. OBJECTIVE: To determine the aetiology, clinical significance and evolution of a group of newborn babies with this type of EEG trace. PATIENTS AND METHODS: We made a retrospective study of fullterm babies in whom burst suppression EEG recordings had been obtained during the neonatal period. RESULTS: We studied 34 patients. In 14 cases the trace was associated with hypoxic ischemic encephalopathy; 4 with meningitis; another 4 with early infantile epileptic encephalopathy (Ohtahara syndrome); 4 cases were attributed to drugs (4 with fentanyl associated in one case with phenobarbitone and in another with midazolam); 2 cases were due to early myoclonic epilepsy; 3 to multiple organ failure; one to non ketotic hyperglycinemia and another to leucinosis. In one patient the aetiology could not be determined. Seven patients died before the age of 6 months. Severe neurological sequelae were seen in all the others except for four cases (3 treated with fentanyl and one case with hypoxic ischemic encephalopathy). CONCLUSIONS: The presence of a burst suppression EEG trace in a neonate makes extensive study to determine the aetiology necessary. Although associated with a worse prognosis, those not treated with piperidine derivatives should be classified separately. Those treated with piperidine derivatives have a good prognosis. PMID- 11727230 TI - [Validation of transcranial Doppler in Mataro, Barcelona]. AB - INTRODUCTION: Transcranial Doppler (TCD) is a new technique which is becoming increasingly used in neurology and is highly dependent on the user. OBJECTIVE: To evaluate the reliability of TCD in our hospital. PATIENTS AND METHODS: During a five month period, all patients who had a cerebral angiogram done, also had DTC within 24 hours. The TCD was reported on before the result of the angiogram was known. Subsequent analysis of the comparison of the results was done by a person who had not done the initial investigations. RESULTS: 49 persons were included in the study. The average time between the two investigations was 12.5 hours. In 5 patients the TCD was inconclusive due to a poor window. In 21 patients both tests were normal. Of the 14 patients in whom there were pathological findings on angiography, correct diagnosis was made on TCD in 12 cases, including those with stenosis of the anterior circulation. The two false negative findings of intracranial stenosis were in a vertebral artery and a posterior cerebral artery. The remaining seven cases were of patients in whom only one investigation showed signs of distal vasculopathy. CONCLUSION: The index of false positives of DTC showing intracranial stenosis was 0. All intracranial stenoses of the anterior circulation were correctly diagnosed. PMID- 11727231 TI - [Diagnose and predict utility of transcranial Doppler in the serious cerebral anoxia]. AB - INTRODUCTION: In the clinical daily practice, exist multiple situations in which could produce phenomenons of serious hypoxia to encephalic level, normally with important repercussions for the patient. OBJECTIVES: Study if after the existence of a situation of serious encephalic hypoxia, any typical pattern in the exploration transcranial Doppler (TCD) exists, and the possible diagnostic and/or prognostic implications could bear. PATIENTS AND METHODS: We presented 10 patients admitted in our Unit of Intensive Care, for clinical situations in which a situation of encephalic hypoxia took place and to which was practiced a TCD like exploration of encephalic flow. We related the discoveries of the DTC, with the clinical situation, the images of the cerebral computed tomography in three of the patients, and the data of the electroencephalogram in the other seven. Like statistical tool, is used the test of square ji, demanding a confidence interval of the 95%. RESULTS: In all the patients, we found a patron TCD, of high speed mean with low Pulsatility Index. Nine patients died, surviving with serious sequels the last patient. CONCLUSIONS: In our series, we could conclude, that the existence of a patron TCD with high speed mean and with low Pulsatility Index, after an episode of brain hypoxia, its associated to a wrong presage, existing a high risk of death of the patient. PMID- 11727232 TI - [Temporal spectral evolution of electroencephalogram. Methodology and applications]. AB - INTRODUCTION: The event related potentials (ERPs) allow to study the evolution of the cerebral electric activity related with the diverse sensorial, motors and cognitive processes. However, the ERPs and the spectral analyses based in Fast Fourier Transform, are not able to show the variations along the time of the spectral content of the electroencephalogram (EEG). OBJECTIVE: The present work will show the necessary methodology to obtain the temporal dynamics of the frequency (TSE) in the EEG, a technique described by Salmelin and Hari (1994) that allows the analysis of the modulations of any spectral band along the time. SUBJECTS AND METHODS: The presented data correspond to the electroencephalographic registration of 20 human subjects during an experiment of visual spatial attention. RESULTS: The modulations of the ERPs differ of the observed by the spectral techniques in latency and topography. Likewise, comparing different spectral techniques is proven that the TSE allows to define with more temporal precision the variations on the spectral content. CONCLUSION: The application of the TSE technique to the same EEG signal that is used for obtaining the ERPs, allows the inclusion of the variable time in the spectral analysis of the EEG, what can be crucial to understand the physiological bases of some neuropathologies. PMID- 11727233 TI - [Electro-oculographic features of persons with vitamin E deficiency]. AB - INTRODUCTION: Vitamin E (VE) deficiency is a very rare condition which may be due to an isolated deficit or be in the context of a malabsorption disorder. OBJECTIVE: To evaluate the findings from an electrooculographic (EOG) point of view of patients with VE deficiency. PATIENTS AND METHODS: We made a retrospective evaluation of all persons with a neurological diagnosis of VE seen in the otoneurological department of our hospital. In all patients an EOG recording was made of the saccadic jerks, and the spontaneous, provoked, positional and opticokinetic (NOC) mystagmus, visual suppression of the vestibulo ocular reflex (VOR) and follow up. RESULTS: We found four cases, with an average age of 9.2 years (range 6 14 years). All the patients but one were male (75%). The EOG findings were related to cerebellar dysfunction (saccadic following) and central nervous system alterations of no value for localization (ataxic following and visual suppression of the VOR). In two patients no anomalies were seen on the EOG recording. CONCLUSIONS: In persons with VE deficiency EOG alterations are not often seen. However, it may be useful to make the recording when it is thought necessary to detect subclinical cerebellar disorders. PMID- 11727234 TI - [Direct carotid-cavernous fistulas: endovascular treatment using a detachable balloon]. AB - INTRODUCTION: Carotid cavernous fistulas are abnormal communications between the cavernous sinus and the internal and/or external carotid artery. They may be spontaneous or acquired. Two basic types are distinguished as a function of the aetiology, clinical features and treatment. The direct type is usually due to trauma while the indirect is of varied aetiology. CLINICAL CASE: We present a series of seven cases of direct carotid-cavernous fistulas, four post-traumatic and three spontaneous, with initial symptoms of ocular involvement and ophthalmoparesia predominantly. All were treated, in our hospital, by embolization with a detachable balloon in the venous variant of the fistula in six cases and by carotid occlusion in one case. Only one severe complication due to distal embolization occurred. CONCLUSIONS: It is necessary to consider the diagnosis of carotid-cavernous fistulas in patients complaining of acute alterations of ocular motility, headache and/or other ocular symptoms, especially when there is a clinical history of head injury. There should be minimal delay between diagnosis and treatment since there is a high risk of hemorrhage and irreversible sequelae caused by alteration of the venous drainage when they are long-standing. The treatment of choice for direct fistulas is by detachable balloon, which has been shown to be superior to other techniques. PMID- 11727235 TI - [Acute encephalitis: bilateral lesions of the basal ganglia]. AB - OBJECTIVE: To refer two children with acute encephalitis and bilateral basal ganglia lesion and its neurological outcome. CLINICAL CASES: Two girls, one of 9 years and the other of 15 months of age were affected by acute encephalitis, bilateral basal ganglia lesion was found on MRI in both children. Abnormal movements (tremor, choreoatetosis) and dystonia were the main symptomatology. In the first girl a mild dystonic posture on her hand and minor bradykinesia was the only found after a 4 year follow up. The other child, after 17 months of the beginning of her disease, still has generalized dystonia and choreoatetosis movements. Control MRI studies, in both patients remain without changes. CONCLUSION: In acute encephalitis, basal ganglia lesion in two children, produced different neurological sequelae, probably related to the age of the presentation of the disease. PMID- 11727236 TI - [Creutzfeldt Jakob disease and hepatic encephalopathy: coexistence in the same patient]. AB - INTRODUCTION: Since detection of a series of cases of a new variant of Creutzfeldt Jakob disease (CJD) in the United Kingdom, prionic diseases are frequently mentioned by the media. The neurologists always consider them in daily clinical practice, although the incidence is relatively low (0.5-2 cases/1,000,000 inhabitants/year for CJD). In Spain, in recent years, we have seen an increased number of probable and possible cases, perhaps due to it always being included in the differential diagnosis, and to the greater use of diagnostic tests such as 14-3-3 protein in cerebrospinal fluid (CSF). There is also a reduction in the number of definite cases, probably due to the reticence to carrying out necropsies. In the differential diagnosis, one must remember structural, infectious, degenerative, toxic, vascular and metabolic alterations, including hepatic encephalopathy (HE). CLINICAL CASE: We report the case of a woman in whom CJD and EH co existed. This made diagnosis difficult. It was finally confirmed on necropsy. PMID- 11727237 TI - [Fibrinolytic treatment of cerebral intraventricular hemorrhage]. AB - INTRODUCTION: Spontaneous cerebral intraventricular hemorrhage (SCIVH) is associated with hydrocephaly which requires ventricular drainage as treatment. Maintenance of adequate debit through the drainage system contributes to reduce morbid mortality. When the drain becomes obstructed, intermittent irrigation is necessary. This causes increased intracranial pressure and risk of infection. We propose the use of recombinant plasminogen tissue activator (r-tPA) in the treatment of SCIVH and associated hydrocephaly. CLINICAL CASES: We gave two patients with SCIVH 4 cc/day of r-tPA intraventricularly, at a concentration of 1 mg/ml over 4 consecutive days. Daily computerized tomography (CT) series were done. In both cases there was complete resolution of the SCIVH, with no complications related to the treatment. The ventricular drainage systems maintained a constant debit, did not become obstructed and required no revision. The patients did not subsequently require permanent ventricular shunts. One patient died of an unrelated septic condition. The other patient, a year later, could walk unaided, had motor dysphasia and on CT had neither intraventricular blood nor hydrocephaly. CONCLUSIONS: The intraventricular administration of r-tPA is a valid therapeutic tool in selected cases. It contributes to lysis of the intraventricular clot, permits permeability of the drainage, and may reduce the incidence of chronic hydrocephaly requiring a permanent ventricular shunt. PMID- 11727238 TI - [Superficial siderosis of the central nervous system: an uncommon cause of spastic paraparesia]. AB - INTRODUCTION: Superficial siderosis of the central nervous system (CNS) is an uncommon neurological condition, characterized clinically by cerebellar ataxia, neurosensorial deafness, anosmia, myelopathy and cognitive deterioration. It is associated with the deposition of haemosiderin in the subpial layers of the brain, cerebellum and spinal cord, following chronic bleeding (often clinically silent) in the subarachnoid space. Histopathologically there is gliosis, neurone loss and demyelination of the CNS. CLINICAL CASE: We present the case of a 60 year old woman with a history of progressive worsening of a disorder of myelopathic type with signs of pyramidal liberation and sphincter incontinence associated with cerebellar ataxia, anosmia and bilateral hypoacusia. Initially she had unsteadiness, frequent falls and weakness of the legs. Her deafness was more obvious during the previous year. On lumbar puncture there was hemorrhagic CSF with increased red blood cells, iron, ferritin and protein. High field encephalic MR showed a hypointense image in T2 which surrounded the fissure of Sylvius, the brain stem, medulla oblongata, cerebellar hemispheres and sulci of the cerebellar vermis, suggestive of hemosiderosis, atrophy of the cerebellar vermis and slight cortical atrophy. Angioresonance of the intracranial vessels showed that there were no signs of aneurysms or vascular malformations. Transcranial Doppler studies were normal. CONCLUSIONS: Superficial siderosis of the CNS should be included in the differential diagnosis of the parethospastic and ataxic syndromes. The extensive study done ruled out any secondary cause such as chronic bleeding secondary to an arteriovenous malformation or bleeding aneurysm. PMID- 11727239 TI - [Lethal nemalinic myopathy and congenital arthrogryposis]. AB - INTRODUCTION: Nemalinic myopathy (NM) is a non progressive congenital disorder of skeletal muscle, characterized by rod like formations present in muscle fibres, whilst congenital multiple arthrogryposis (CMA) is a syndrome characterized by contractures and articular rigidity which may be due to many causes, including NM. CLINICAL CASE: A boy of 0 h of premature life, with CMA was assessed on the suspicion of neuromuscular disease with normal preliminary studies. Muscle biopsy eventually showed the typical rod like formations of NM. The clinical course was unsatisfactory and he died at the age of five months from respiratory problems. CONCLUSIONS: Diagnosis of NM requires a high index of suspicion and muscle biopsy. At the present time genetic counselling cannot be given after a sporadic case. PMID- 11727240 TI - [Use of the technology of the genetic chip to study of the nitric oxide synthase]. AB - INTRODUCTION: The end of our century has been characterized by a quick scientific and technological growth. Among other, the brain, the human genome project and the computer science, they are some of the numerous fields that characterize this revolution. DEVELOPMENT: In the following work, we will show as these three fields of the knowledge have converged in a new area of the science denominated technology of the gene chip or microarrays. Although this methodology would allow the study of thousands of genes in one chip, in the following work we will center ourselves in its potential application for the study of the enzyme nitric oxide synthase (NOS), which is responsible for the production of the nitric oxide (NO). The technical base of this system is to be able to synthesize chains of DNA in a chip, which will be hybridized with the sample of interest that in turn are usually marked with fluorescent nucleotides. The results are analyzed using a sophisticated image analysis system. The relative estimate of the quantity of existent messenger in each sample will come determined by the intensity of the fluorescence, and for its comparison with internal controls. The discovery of at least three genes that are coded for NOS, as well as that of numerous allelic variations will allow a more exhaustive study of this molecule, which has been recognized by their vital importance. CONCLUSION: In this review we will put in perspective the utility of this technology in the genetic study of the enzyme NOS, and its utility for the understanding of the pathophysiology of some neurological illnesses. PMID- 11727241 TI - [Neuropsychiatric disorders in multiple sclerosis]. AB - OBJECTIVE: Neuropsychological investigations over the past 20 years have shown the high prevalence of cognitive dysfunction in multiple sclerosis, although there are still large gaps. In this article we review current concepts regarding the different types of neuropsychiatric disorders, their physiopathology, diagnosis and treatment. DEVELOPMENT: We present a classification of the most important types of cognitive and psychiatric alterations in multiple sclerosis, and their physiopathology. We review the methods used in diagnosis, follow up and assessment of these patients. Finally, we analyze the current treatment available for each of these disorders. CONCLUSIONS: Although considerable advances have been made in understanding the neuropsychological disorders occurring in multiple sclerosis, there are still many questions to be answered. There is a need for a unified, agreed, validated battery of neuropsychological tests for the study of specific cognitive deficits in multiple sclerosis so that results may be compared. There is also a lack of biological markers which would help in diagnosis and prognosis. Finally the design and carrying out of clinical trials in the search for specific, effective drugs for the treatment of neuroconductual disorders of multiple sclerosis is necessary. PMID- 11727242 TI - [Cerebral mechanisms of mathematical thinking]. AB - OBJECTIVE: To suggest a cerebral map of elementary mathematical thinking, and integrate the most relevant findings from neuropsychology with those from cerebral imaging techniques and cognitive behavior experiments. DEVELOPMENT: Firstly we describe investigations into our numerical sense and the way in which numerical information is represented in the human brain. Then, using a multidisciplinary approach, we present the results of different studies of Gerstmann's syndrome, regarding the relation between numerical ability and other cognitive skills; the different participation of the cerebral hemispheres and the special implication of the parietal lobe in mathematical tasks. CONCLUSIONS: Different cerebral regions are involved in doing mental arithmetic, however simple. This makes one think more in terms of cerebral circuits than in a phrenological idea which would assign the responsibility for arithmetical calculations to a specific region. The similarity between the results analysed leads us to the conclusion that one region is particularly involved in understanding numbers, namely the inferior part of the parietal lobe. Different neuronal circuits are used depending on the type of task to be performed. Finally we describe the most relevant models for the processing of numbers which have been developed during the study. PMID- 11727243 TI - [Alcohol and stroke: a controversial association]. AB - OBJECTIVE: Although an increasing quantity of information concerning risk factors for stroke have been developed lately, the relationship between alcohol consumption and cerebral ischemic infarction, unlike at coronary sites, remains unclear. This review highlights the main pathogenic mechanisms of alcohol in the development of ischemic stroke, as it intends to be an update of this possible association. DEVELOPMENT: A critical literature review has been performed regarding this association to study its pathogenic mechanisms and the possible confounding factors present in the current design of epidemiological reports. CONCLUSIONS: There is an increasing evidence about the effect of light to moderate alcohol consumption and prevention of ischemic stroke. On the other hand it seems that heavy drinking, and particularly binge drinkers, are exposed to a higher risk for cardioembolic stroke. The growing incidence of acute heavy alcohol consumption highlights the necessity of considering a cardioembolic source when managing ischemic stroke, specially in young adults. PMID- 11727244 TI - [Relation of the cerebellum with cognitive function: neuroanatomical, clinical and neuroimaging evidence]. AB - INTRODUCTION: Published studies in the last decades, employing a variety of methodologies, suggest that the cerebellum function is not only limited to motor control but also has an important role in cognition and emotion. DEVELOPMENT: In the present paper we review the main neuroanatomical and functional neuroimaging studies supporting the idea that the cerebellum has a role in non motor behavior. On the other hand, we expound the results of the neuropsychological studies of patients with cerebellar lesions. The deficits showed after cerebellar lesions include disturbances in executive functions, learning, memory, attention, visuo spatial functions, language and personality change. Some studies propose that the cerebellum, through its afferent and efferent circuits with the cerebrum, perform a global function capable to influencing motor behavior, in cognitive functions and in emotion. A fault in this function would cause the behavioral deficits shown in patients with cerebellar lesions. CONCLUSION: The evidence available in the anatomical, functional and clinical studies support the conclusion that the cerebellum participates not only in motor functions, but also in cognitive functions and in emotion. PMID- 11727245 TI - [Bilateral carotid occlusion as a form of presentation of an aneurysm of the aortic arch in a 39 year old man]. PMID- 11727246 TI - [Type 1 neurofibromatosis, epilepsy and disorders of neuronal migration]. PMID- 11727247 TI - [Surgery for cervical spondylosis using an anterior approach]. PMID- 11727248 TI - [Overdiagnosis in the epidemic of neuropathy in Cuba]. PMID- 11727250 TI - [Brachial plexopathy in varicella of the adults. A case report]. PMID- 11727251 TI - Curricular trends in instruction of pathology: a nationwide longitudinal study from 1993 to present. AB - Medical schools throughout the United States continue to respond to various external and internal challenges and make modifications in their curricula. Responses obtained from 66% (n = 83) of schools on a longitudinal survey conducted during the year 2000 to study trends in instruction of pathology over a 7-year period (ie, for classes entering 1993 to 1999) indicate the following. There have been steady shifts in instruction of systemic and clinical pathology from discipline-based courses to integrated formats from 1993 to 1999. The degree of integration with other disciplines varies among schools, and may take one or more of the following formats: joint course (pathology and another discipline); multidisciplinary systems course; a combination of pathology course and another integrated course; and completely integrated in the form of problem-based or case based small group discussion. Presently, at least some degree of integration occurs in 51% of schools for instruction of systemic pathology and 65% for clinical pathology, up from 31% and 48%, respectively, in 1993. Although there has been an increased trend toward integration in instruction of general pathology as well, it is still taught predominantly in a discipline-based manner in the vast majority of schools. Although often difficult to identify with certainty, the best estimates indicate that the overall scheduled length of pathology instruction time has stabilized over the years; the mean total for the entering classes of 1999 was 196 hours versus 201 hours for the classes entering in 1993. However, internal rearrangements of time for various components of delivery of instruction continue. The lecture remains unchanged as an important mode comprising the largest component of pathology curriculum time (53% in 1999, 52.2% in 1993) during the 7-year period examined. The mean traditional laboratory instruction time has decreased slightly from 27% in 1993 to 24% in 1999. This decrease may be accounted for by a shift toward use of laboratory materials in various other formats and venues not included in the traditional laboratory instruction (eg, small group sessions, clinicopathologic correlation conferences, study of kodachrome slides, and computer programs). The use of electronic educational resources has increased remarkably, but for the most part it is not measurable because of the lack of any designated hours. Because pathology instruction occurs exclusively or primarily during year 2 in most schools, the classes entering in 1999 would have studied it during 2000 and 2001, which means the present study provides the most updated curricular trends at this time. PMID- 11727252 TI - The teaching of anatomic pathology in England and Wales: a transatlantic view. PMID- 11727253 TI - Expression of p53-related protein p63 in the gastrointestinal tract and in esophageal metaplastic and neoplastic disorders. AB - p63 is a p53-related DNA-binding protein that helps regulate differentiation and proliferation in epithelial progenitor cells. Its expression has never been evaluated in the human gastrointestinal tract. The aim of this study was to evaluate the expression of p63 in the esophagus and related metaplastic and neoplastic disorders to gain insight into the pathogenesis of these processes. Of particular interest was the expression of p63 in Barrett esophagus (BE) and in BE associated multilayered epithelium. Multilayered epithelium has been postulated to represent an early precursor to the development of BE primarily because it shares morphologic and immunophenotypic features of both squamous and columnar epithelium, and has been shown prospectively to be highly associated with BE. Routinely processed mucosal biopsy or resection specimens that contained normal esophageal squamous epithelium (n = 20), squamous dysplasia (n = 4), squamous cell carcinoma (n = 7), BE (n = 10), BE-associated multilayered epithelium (n = 13), esophageal mucosal gland ducts (n = 10), BE-associated dysplasia (n = 12), and BE-associated adenocarcinoma (n = 7) were immunostained for p63 to determine the extent and location of staining. p63 staining was compared with the staining patterns observed for p53, Ki 67 (proliferation marker), and cytokeratins (CKs) 13 (squamous marker), 14 (basal squamous marker), 8/18 (columnar marker), and 19 (basal/columnar marker). Expression of p63 messenger RNA (mRNA) isoforms was also analyzed by reverse-transcription polymerase chain reaction of freshly isolated tissues. In the normal esophagus, p63 was expressed in the basal and suprabasal layers of the squamous epithelium and in basal cells that line the mucosal gland ducts but was negative in all other epithelia of the gastrointestinal tract, including the stomach, small intestine, and colon. Similarly, p63 was not expressed in BE, but it, was present in the basal layer of multilayered epithelium in 9 of 13 cases (69%). p63-positive cells in multilayered epithelium and in the mucosal gland duct epithelium were positive for CK8/18 (100%) and CK13 (67% and 30%, respectively) and negative for CK14 (0%), in contrast to p63 positive cells in squamous epithelium, which were positive for CK14 and CK13 (100%) but negative for CK8/18. In neoplastic tissues, p63 was diffusely expressed in all cases of esophageal squamous cell dysplasia and carcinoma but was negative in all cases of esophageal and colorectal adenocarcinoma. The DeltaN isoform of p63 mRNA predominated in all benign and neoplastic squamous tissues examined. p63 may represent a marker of 2 distinct epithelial progenitor cells (basal squamous epithelium and gland duct epithelium) in the esophagus. P63 is upregulated in squamous neoplastic conditions and in this manner may play a role in squamous carcinogenesis. These data also indicate that multilayered epithelium is phenotypically similar to, and may share a lineage relationship with, mucosal gland duct epithelium. PMID- 11727254 TI - Expression of microtubule-associated protein tau by gastrointestinal stromal tumors. AB - The purpose of this work was to study the expression in gastrointestinal stromal tumors (GISTs) of various antigens, including the protein tau associated with enteric neuronal differentiation; to compare their expression with that of c-kit, known to be associated with interstitial cell of Cajal differentiation; and to correlate their expression with the observation of ultrastructural features of gastrointestinal autonomic nerve tumors. Twenty-six GISTs of the stomach and 16 GISTs of the small bowel were included in the study group. Thirty-five tumors served as controls. Tissue sections were immunostained with vimentin, CD34, desmin, specific smooth muscle actin, S100 protein, neuron-specific enolase, PGP9.5, neurofilament, bcl-2 oncoprotein, synaptophysin, chromogranin A, c-kit, and tau. Twenty-one of these tumors were also analyzed ultrastructurally. Of the 42 GISTs, 28 were predominantly spindled, 7 were predominantly epithelioid, and 7 were a mixture of epithelioid and spindle cells. Ten primary GISTs were classified as benign, 9 as borderline, and 23 as malignant. Metastatic dissemination was present at primary surgery in 1 case and eventually developed in 6 patients. Six disease-related deaths were counted. In normal submucous and myenteric plexuses of stomach and small bowel, ganglion cell bodies and nerve fibers strongly expressed tau. Twenty (76.9%) GISTs of the stomach and 12 (75%) of the small bowel expressed tau. Tau often showed intense, diffuse staining patterns in both spindled and epithelioid tumors. Ten (100%) of the 10 benign GISTs, 7 (77.8%) of the borderline GISTs, and 15 (65.2%) of the 23 frankly malignant GISTs expressed tau. Thirty-six GISTs expressed at least 2 different neuronal markers. A coexpression of the neuronal markers and c-kit was observed in 90% of GISTs. The expression of tau was observed in 12 of the 15 GISTs with dense core granules, considered as the definitive finding for a diagnosis of gastrointestinal autonomic nerve tumors. Ten of these also expressed c-kit; 9 were malignant. Tau also immunostained other intra-abdominal tumors, including neuroendocrine carcinomas, paragangliomas and desmoplastic round cell tumors. This immunohistochemical study shows that GISTs are specific tumors of the digestive tract and are nearly always characterized by simultaneous neuronal and interstitial cell of Cajal differentiation. Although the loss of tau expression is observed only in borderline and malignant tumors, its prognostic value is not clear cut. PMID- 11727255 TI - Mast cell distribution and activation in chronic pancreatitis. AB - Chronic pancreatitis (CP) is characterized by mononuclear inflammatory cell infiltration and replacement of the destroyed parenchyma by fibrous tissue. Recently, mast cells have been implicated in chronic inflammatory processes with fibrous tissue deposition. Therefore, the number and distribution of mast cells and their state of activation were evaluated in 12 normal specimens and in 46 specimens of CP with different causes (alcoholic, tropical, and idiopathic). Furthermore, the presence of stem cell factor (SCF), the main mast cell growth factor, and of its receptor, c-kit, was also assessed. In CP tissues, mast cells were localized both in the fibrotic areas and in the residual acinar parenchyma. The total number of mast cells was significantly higher in CP than in the normal pancreas (P < .0001) and correlated positively with the extent of fibrosis and the intensity of inflammation. Immunoglobulin E (IgE)-dependent mast cell activation was higher in CP than in the normal pancreas. No differences in mast cell number or IgE positivity were found among the 3 causes of CP. SCF-and c-kit immunoreactive mast cells were mostly localized in fibrous tissue and around regenerating ducts, which were also positive for c-kit but were negative for SCF. These results suggest that mast cells, activated by an IgE-dependent mechanism and/or by an SCF-c-kit autocrine loop, are a relevant component of the inflammatory infiltrate in CP, independent of the underlying cause. Their localization near degenerating acini and regenerating ducts might indicate that they play a crucial role in tissue destruction and remodeling in CP. PMID- 11727256 TI - Immunohistochemical expression of transforming growth factor alpha and epidermal growth factor receptor in pancreatic endocrine tumors. AB - Coexpression of transforming growth factor alpha (TGF-alpha) and its receptor epidermal growth factor receptor (EGFR) is known to be associated with aggressive biologic behavior and adverse clinical outcome in a variety of tumors, including pancreatic adenocarcinomas. However, very little information is currently available as to whether this is true of pancreatic endocrine tumors (PETs) as well. Thirty-five PETs were retrospectively studied for immunohistochemical expression of TGF-alpha, the intracellular and extracellular domains of EGFR, and various hormonal secretory products. Proliferative activity was additionally studied (in 20 cases only) using the MIB-1 antibody. Thirty-one (89%) of 35 tumors were reactive for 1 or more of the peptide hormones tested; 22 (63%) tumors were positive for TGF-alpha; and 23 (65%) were positive for the intracellular and/or extracellular domain of EGFR. Based on their TGF-alpha and EGFR expression, these tumors could be classified into 4 groups. Of the 10 tumors in group I (positive for TGF-alpha and the complete EGFR molecule), 3 were malignant, 6 were >2 cm in diameter, 5 were functional, and 1 had a proliferative index of >40%. The 12 tumors in group II (positive for TGF-alpha but negative for the intracellular and/or extracellular domain of EGFR) included 4 malignant tumors, 4 PETs >2 cm in diameter, 8 functional, and 1 with a proliferative index of >40%. The 7 PETs in group III (positive for the intracellular/extracellular domain of EGFR alone) included 3 malignant tumors, 3 PETs >2 cm in diameter, and 3 functional tumors. The 6 tumors in group IV (completely negative for both TGF alpha and EGFR) included 4 malignant tumors, 3 PETs >2 cm in diameter, and 4 functional lesions. Therefore, immunohistochemical expression of TGF-alpha and EGFR, either alone or in concert, shows no correlation with size, functional status, secretory profile, or biologic behavior and hence cannot be used as a marker of malignancy in this group of tumors. PMID- 11727257 TI - Expression of CD44 standard and isoforms V3 and V6 in uterine smooth muscle tumors: a possible diagnostic tool for the diagnosis of leiomyosarcoma. AB - Alterations of CD44 proteins, a family of cell adhesion molecule, have been linked with tumorigenesis, carcinogenesis, and prognosis in various neoplasms. Our aims were to evaluate and compare CD44 isoforms expression patterns in normal myometrium, uterine leiomyomas, and leiomyosarcomas and to correlate CD44 expression with clinicopathologic parameters. Fresh (n = 15) and formalin-fixed, paraffin-embedded (n = 76) tissues samples of myometrium, leiomyomas, and leiomyosarcomas were used for immunoblotting and immunohistochemistry, respectively. Semiquantitative evaluation was made after immunostaining. Monoclonal antibodies were used. By immunoblotting in myometrium and leiomyomas samples, we observed a band at 85 kd, corresponding to the apparent molecular weight of CD44s, and bands at 140 kd with the monoclonal antibodies against CD44v3 and CD44v6. In leiomyosarcomas, CD44s and CD44v6 were detected, but not CD44v3. By immunohistochemistry, decreased CD44s expression was found in leiomyomas and leiomyosarcomas (73.9% +/- 16.6% and 82.1% +/- 20.7%, respectively) compared with myometrium (97.3% +/- 6.2%; P < .0001). No CD44v6 staining was detected in myometrium, leiomyomas, and leiomyosarcomas. No CD44v3 expression was detected in leiomyosarcomas, whereas myometrium and leiomyomas expressed CD44v3. For the diagnosis of leiomyosarcoma, the absence of CD44v3 staining had a sensitivity, specificity, and positive and negative predictive values of 100%. In patients with recurrence of leiomyosarcomas, CD44s expression was decreased (P = .03). We conclude that CD44s immunostaining in leiomyosarcomas may have prognostic significance. The loss of CD44v3 expression could be used as a putative diagnostic tool for uterine leiomyosarcomas. PMID- 11727258 TI - MUC4 expression is increased in dysplastic cervical disorders. AB - The female uterine cervix has 2 characteristic populations of epithelial cells: the endocervix is composed by mucus-secreting cells that express several mucin genes, and the exocervix has a typical stratified squamous epithelium and does not express secreted mucins. Among human mucin genes, the MUC4 sequence has a transmembrane domain, and its molecular structure suggests that it has a protective role and also may be implicated in intracellular signalling. The aim of this study is to analyze whether changes in the expression of MUC4 can be detected associated with the squamous dysplastic transformation of exocervical epithelium. MUC4 expression has been analyzed by immunohistochemistry, Western blotting, and in situ hybridization. Using immunohistochemical techniques, MUC4 is found in normal endocervix (n = 11) and is absent or only focally detected in the normal stratified cervical epithelium (n = 18). In samples from squamous metaplasia (n = 9), MUC4 is variably expressed (10% to 50% positive cells), whereas MUC4 is strongly detected in dysplastic cervical epithelia. The greatest number of positive cells is found in samples with moderate and severe dysplasia in which MUC4 is detected in 100% of the analyzed samples (n = 16). These results have been confirmed by Western blotting and by detection of MUC4 transcripts using in situ hybridization. The present data suggest that MUC4 is activated during the process of squamous dysplastic transformation and may be used as a marker for this pathologic process. PMID- 11727259 TI - Heterogeneous genetic alterations in ovarian mucinous tumors: application and usefulness of laser capture microdissection. AB - Histologic observation of ovarian mucinous tumors suggests that there is a multistep transition through the accumulation of genetic alterations. We analyzed loss of heterozygosity (LOH) and replication error (RER) on TP53 and D17S855 as well as K-ras point mutations of the heterogeneous histologic areas of the same tumor in 26 cases of ovarian mucinous tumor. The laser capture microdissection (LCM) technique has been applied to the study of K-ras point mutation in 10 cases. As for genetic alterations for LOH or RER on TP53 and D17S855, 2 (1 borderline tumor and 1 carcinoma) of 14 cases and 4 (1 borderline tumor and 3 carcinomas) of 12 cases, respectively, showed genetic heterogeneities in different histologic areas. Six (2 borderline tumors and 4 carcinomas) of 18 cases showed heterogeneity of K-ras point mutation in the different histologic areas of the same tumor, and 5 (1 cystadenoma with Brenner tumor component, 2 borderline tumors, and 2 carcinomas) of 10 cases showed heterogeneous K-ras mutation pattern in the same tumor when the LCM technique was used. Atypical areas tended to show K-ras point mutations frequently. Out of 3 cases of mixed mucinous cystadenoma and Brenner tumor, 1 case showed K-ras point mutation in the Brenner tumor area but not in the area of mucinous cystadenoma. These preliminary results suggest that a subset of ovarian mucinous tumors occur through multistep carcinogenesis and show that LCM is useful for molecular pathologic studies. PMID- 11727260 TI - Clinical significance of cell proliferation, microvessel density, and CD44 adhesion molecule expression in renal cell carcinoma. AB - Renal cell carcinoma (RCC) is known to have a wide variation in clinical outcome despite the use of conventional prognostic factors, such as staging or grading. A better knowledge of the biologic aggressiveness of RCC could facilitate the selection of patients at high risk of tumor progression. The aim of this study was to determine if use of measurements of vascular density, cell proliferation, and cell adhesion could better predict the biologic behavior of RCC. We immunohistochemically analyzed CD34, Ki-67, and CD44H expression on formalin fixed, paraffin-embedded tissues from 73 RCCs for quantifying microvessel density (MVD), Ki-67 labeling index (LI), and CD44H LI, respectively. Univariate cancer specific survival analysis showed that tumor stage (P < .01), tumor size (P < .001), nuclear grade (P < .01), metastasis (P < .001), MVD (P < .03), Ki-67 LI (P < .001), and CD44H LI (P < .0001) were predictors of tumor-related death. There was a statistical correlation between CD44H LI and both Ki-67 LI (r' = .3) and MVD (r' = -44). Ki-67 LI (P < .04) and CD44H LI (P < .02), as well as metastasis (P < .008), emerged as independent predictors of cancer-specific survival in multivariate analysis in patients with metastases (P < .04 and P < .02, respectively) and in patients without metastases (P < .006 and P < .00001, respectively). Our study suggests that vascular density, cell proliferation, and cell adhesion represent a complex tumor-host interaction that may favor progression of RCC. Cell proliferation and CD44H expression appear to be powerful markers to identify patients with an adverse prognosis. PMID- 11727261 TI - The expression of E-cadherin and catenins in sporadic colorectal carcinoma. AB - The E-cadherin/catenin complex plays a major role in epithelial cell-cell adhesion. Immunohistochemical studies have highlighted perturbation in the expression and distribution of E-cadherin and catenins in sporadic colorectal neoplasms. In this study, we compared the expression of E-cadherin and catenins (alpha-, beta-, and gamma-catenin) in 30 sporadic colorectal carcinomas with that in the adjacent nonneoplastic mucosa and assessed whether any perturbation in the level of expression occurred at the messenger RNA (mRNA) or protein level. We also compared the expression of E-cadherin and catenins in 13 lymph node deposits and the primary tumors. Immunohistochemistry was used to study the level of expression and cellular distribution of E-cadherin and catenins. Levels of mRNA were studied by in situ hybridization. E-cadherin and catenin immunoreactivity was increased with cytoplasmic accumulation in more than 85% of the neoplasms. There were marked increases in the levels of mRNA in the carcinomas compared with the nonneoplastic mucosa. Nuclear localization of beta-catenin was higher at the invasive margin of some tumors, but expression of E-cadherin and catenin transcripts in the lymph node deposits showed no consistent relationship to that in the primary tumors. PMID- 11727262 TI - H-, K-, and N-ras gene mutation in atypical fibroxanthoma and malignant fibrous histiocytoma. AB - Atypical fibroxanthoma (AFX) which is histologically similar to malignant fibrous histiocytoma (MFH), occurs in the sun-exposed skin. The presence of mutations at codons 12 and 13 of the H- and K-ras genes and in exons 1 and 2, which include codons 12, 13, and 61, of the N-ras gene was studied in 8 cases of AFX and 8 cases of storiform-pleomorphic-type MFH using polymerase chain reaction (PCR) restriction fragment length polymorphism and PCR-single-conformation polymorphism. Two of the 8 cases of MFH showed ras mutations in the H-ras gene at codon 12 (GGC-AGC) and in the K-ras gene at codon 13 (GGC-GAC). H- and K-ras gene mutations were not seen in any of the cases of AFX (0 of 8). N-ras gene mutation was not detected in either the AFX (0 of 8) or MFH (0 of 8) cases. In conclusion, although the number of cases in this study was small, H- and K-ras genes were present in some of the MFH cases and accordingly may play an important role in the pathogenesis of MFH. In addition, the finding that H-, K-, and N-ras gene mutations are not present in AFX may indicate why AFX has a more favorable behavior than MFH. PMID- 11727263 TI - Contribution of the microvessel network to the clonal and kinetic profiles of adrenal cortical proliferative lesions. AB - Monoclonal adrenocortical lesions have been characterized by an inverse correlation between proliferation and apoptosis, and polyclonal lesions show a direct correlation. Their relationship with the vascular pattern remains unknown in adrenocortical nodular hyperplasias (ACNHs), adenomas (ACAs), and carcinomas (ACCs). We studied 20 ACNHs, 25 ACAs, and 10 ACCs (World Health Organization classification criteria) from 55 women. The analysis included X-chromosome inactivation assay (on microdissected samples), slide and flow cytometry, and in situ end labeling. Endothelial cells were stained with anti-CD31, and the blood vessel area and density were quantified by image analysis in the same areas. Appropriate tissue controls were run in every case. Regression analyses between kinetic and vascular features were performed in both polyclonal and monoclonal lesions. Polyclonal patterns were observed in 14 of 18 informative ACNHs and 3 of 22 informative ACAs, and monoclonal patterns were seen in 4 of 18 ACNHs, 19 of 22 ACAs, and 9 of 9 ACCs. A progressive increase in microvessel area was observed in the ACNH-ACA-ACC transition but was statistically significant between benign and malignant lesions only (191.36 +/- 168.32 v 958.07 +/- 1279.86 microm(2); P < .0001). In addition, case stratification by clonal pattern showed significant differences between polyclonal and monoclonal benign lesions; 6% of polyclonal and 57% of monoclonal lesions had microvessel area >186 microm(2) (P = .0000008). Monoclonal lesions showed parallel trends (but with opposite signs) for microvessel area and density in comparison with proliferation and apoptosis, whereas polyclonal lesions showed inverse trends. In conclusion, the kinetic advantage of monoclonal adrenal cortical lesions (increased proliferation, decreased apoptosis) is maintained by parallel increases in microvessel area and density. PMID- 11727264 TI - Expression of multidrug resistance 1 and glutathione-S-transferase-Pi protein in nasopharyngeal carcinoma. AB - Radiotherapy is the modality of choice for the treatment of nasopharyngeal carcinoma (NPC). However, systemic chemotherapy has recently been found to play an increasing role in the treatment of advanced or metastatic disease. The status of drug resistance gene expression that has crucial impact on chemotherapy has not been fully addressed for patients with NPC. In this study, we examined the expression of multidrug resistance 1 (MDR-1) and glutathione-S-transferase-Pi (GST-Pi) in primary, recurrent, and metastatic NPC using results of immunohistochemical examinations. The results were correlated with the expression of Epstein-Barr virus (EBV) latent protein, latent membrane protein 1 (LMP1), and clinicopathologic features, including stage, histopathologic types, and survival rates. MDR-1 protein expression was detected in 18 (12.6%) of 143 patients with primary NPC, 14 (32.6%) of 43 with recurrent NPC, and O (0%) of 20 with metastatic NPC, whereas 83 (58%) of 143 patients with primary NPC, 30 (69.8%) of 43 with recurrent NPC, and 13 (65%) of 20 with metastatic NPC expressed GST-Pi. EBV-LMP1 was expressed in 59 (41.3%) of 143 patients with primary NPC, 23 (53.5%) of 43 with recurrent NPC, and 9 (45%) of 20 with metastatic NPC. Simultaneous expression of MDR1 and GST-Pi was observed in 13 (72.2%) of 18 patients with primary NPC and 12 (85.7%) of 14 with recurrent NPC. The expression of LMP1 was detected in only 6 of the 13 patients with primary NPC and 6 of the 12 with recurrent NPC. We concluded that the expression of GST-Pi was more frequent in NPC tumor tissues than the expression of MDR-1. The expression of MDR-1 correlated with clinicopathologic features of primary NPC, including the histopathologic types and survival rates, but not with disease stage. The expression of GST-Pi did not correlate with clinicopathologic features. The expression of MDR-1 and GST-Pi did not correlate with expression of EBV-LMP1 for patients with NPC. PMID- 11727265 TI - Immunoexpression of neurofibromin, S-100 protein, and leu-7 and mutation analysis of the NF1 gene at codon 1423 in osteofibrous dysplasia. AB - The NF1 (neurofibromatosis type 1, or von Recklinghausen disease) gene, is a tumor-suppressor gene, and its product, neurofibromin, down-regulates ras protein by its guanosine triphosphatase-activating protein (GAP)-related domain. Osteofibrous dysplasia (OFD) is characterized by fibroblast-like spindle cells and osseous tissue and is generally seen in the tibia or fibula during childhood. The precise nature of OFD remains controversial. Cosegregations of OFD and NF1 have been reported, and it has been surmised that OFD is associated with the NF1 gene. We studied the expressions of NF1 gene product (neurofibromin) and so called Schwann cell markers (S-100 protein, Leu-7) in 17 cases of OFD immunohistochemically. Ten cases of fibrous dysplasia (FD) were also used for the purpose of comparison. Five OFD and 7 FD cases were analyzed for NF1 gene mutation at codon 1423, which is a GAP-related domain, by single-strand conformation polymorphism. Fibroblast-like cells of OFD showed the expression of neurofibromin (5 of 17), S-100 protein (9 of 17), and Leu-7 (5 of 17), and those of FD did not show these expressions, with the exception of 1 case that showed Leu-7 expression. Regarding the OFD cases, significant correspondence was found between cases showing expression of neurofibromin and S-100 protein, between cases showing expression of neurofibromin and Leu-7, and between cases showing expression of S-100 protein and Leu-7 (P < .01). NF1 gene mutation at codon 1423 was not detected in either the OFD (0 of 5) or FD (0 of 7) cases. These results seem to suggest the possible involvement of neurofibromin in the development of OFD, which is associated with the expression of Schwann cell markers (S-100 protein and Leu-7). Furthermore, NF1 gene mutation at codon 1423 did not seem to be related to OFD. PMID- 11727266 TI - Carcinoid tumors of the duodenum and ampulla of vater: a clinicomorphologic, immunohistochemical, and cell kinetic comparison. AB - Carcinoid tumors of the ampulla of Vater (ACs) differ from duodenal carcinoid tumors (DCs). A search for AC and DC was made between 1980 and 2000. The clinicopathologic features and follow-up were assessed. Immunohistochemistry for panneuroendocrine markers, hormone products, proliferating cell nuclear antigen (PCNA), Ki- 67, p21(cip1), and p27(kip1) were performed. A blind proliferative index counting 500 cells was made. Differences were contrasted using the Fisher exact and 2-sided Student t test. Five ACs and 8 DCs were identified in 9 women and 4 men with median ages of 59 and 64 years and mean tumor diameters of 1.6 and 1.85 cm, respectively. All patients with AC presented jaundice, and most patients with DC were asymptomatic (P = .047). Metastases were present in 4 ACs and 1 DC (P =.03). Tumor cells expressed synaptophysin and chromogranin in 60% of ACs and in 100% and 87% of DCs. Gastrin was expressed in 75% of DCs and 20% of ACs (P < .05). The mean value for PCNA index was 4.0% in ACs and 3.2% in DCs, and mean values for Ki-67 were 12.2% and 10.2%, respectively (P = NS). Expression of p21(cip1) and p27(kip1) was observed in 40% of ACs and 37.5% and 12.5% of DCs. Three of 5 patients with AC died of the disease within an average of 11 months, and none of the patients with DC had died at 103 months of follow-up. The more aggressive behavior of ACs is not associated with higher proliferative indices or with different expression of cell cycle inhibitors. PMID- 11727267 TI - Expression and regulation of hemeoxygenase 1 in healthy human lung and interstitial lung disorders. AB - Hemeoxygenase 1 (HO-1) has been implicated in the protection of lung tissue against exogenous oxidant exposure. However, the expression and cellular distribution of HO-1 in human lung continue to be poorly characterized. The localization of HO-1 was studied in histopathologically healthy lung from nonsmoking patients with a carcinoid or other lung tumor (5 cases), pulmonary sarcoidosis (13 cases), and chronic interstitial pneumonias (9 cases, usual interstitial pneumonia; 10 cases, desquamative interstitial pneumonia). Immunostaining was graded from 0 (no immunoreactivity) to +++ (intense immunoreactivity). In healthy lung, HO-1 was localized to alveolar macrophages with reactivity varying from moderate to intense, and in bronchial epithelium, alveolar epithelium, endothelium, and interstitium, the immunoreactivity was not detectable or was very low. Sarcoidosis and interstitial pneumonias showed intense HO-1 immunoreactivity in alveolar macrophages in most of the cases and weak to intense immunoreactivity in the granulomas of sarcoidosis. The immunoreactivity of interstitium was negative or weak in the fibrotic areas of the lung and also in the samples of bronchoalveolar lavage fluid obtained from the patients with UIP. Western blotting indicated that HO-1 is up-regulated by exposure of monocytes to formylated peptide, fMLP, which causes respiratory burst in the cells, and that inhibition of HO-1 by tin protoporphyrin potentiates the injury of fMLP-exposed cells. In conclusion, these data show differential distribution of HO-1 in human lung cells and strongly suggest the importance of HO-1, especially in the defense of alveolar macrophages in normal human lung and in the inflammatory, but not in the fibrotic, stage of interstitial lung disorders. PMID- 11727268 TI - Epstein-Barr virus-associated extranodal NK/T-cell lymphoma, nasal type of the hypopharynx, in a renal allograft recipient: case report and review of literature. AB - Posttransplant lymphoproliferative disorders (PTLPDs) are predominantly B-cell lymphoproliferations, whereas a T-cell origin is rarely observed. In contrast to B-cell PTLPD, T-cell PTLPDs show an inconsistent association with Epstein-Barr virus (EBV). Until now, only 13 cases of EBV-associated T-cell PTLPDs have been reported. We describe a case of an EBV-associated T-cell PTLPD in a renal allograft recipient 2 years after transplantation. Histologic examination showed medium- to large-sized lymphoid cells with an angiocentric growth pattern and necrosis. The atypical cells showed a CD2+, CD3epsilon+, CD7+, CD43+, CD45R0+, CD56+, and CD4-, CD5-, CD8- betaF1- phenotype with expression of the latent membrane protein (LMP)-1 of EBV. In addition, EBV-specific RNAs (EBER 1/2) were identified by in situ hybridization. Molecular analysis of the T-cell receptor (TCR) gamma chain by polymerase chain reaction (PCR) showed a polyclonal pattern. The morphologic, immunohistochemical, and molecular findings were consistent with a diagnosis of an EBV-associated extranodal natural killer (NK)/T-cell non Hodgkin lymphoma (NHL) of nasal type. To our knowledge, this is the first reported case of this rare entity in the posttransplant setting. PMID- 11727269 TI - Hodgkin's lymphoma of T-cell type: clonal association with a CD30+ cutaneous lymphoma. AB - The derivation of Reed-Sternberg cells in Hodgkin's lymphoma has been a subject of great interest. In most cases, Reed-Sternberg cells seem to be derived from germinal center B cells. In few sporadic cases, a T-cell origin has been shown. This article supports the concept of a T-cell derivation for rare cases of Hodgkin's lymphoma and provides evidence of a novel mechanism of pathogenesis from chronic inflammation in the skin. PMID- 11727270 TI - Mutation of von Hippel-Lindau tumor suppressor gene in a sporadic endolymphatic sac tumor. AB - Endolymphatic sac tumor (ELST) is a low-grade adenocarcinoma of the temporal bone that is presumed to originate from the endolymphatic system. Although ELSTs are extremely rare in the general population, a significant number of studies have documented the occurrence of ELST among patients with von Hippel-Lindau (VHL) disease. Because of the rarity of the tumor, however, few cases of ELST have been analyzed for mutations of the VHL tumor suppressor gene. In this study, we reported a Japanese male patient with sporadic ELST, along with a molecular genetic analysis of the VHL gene. The light microscopic and immunohistochemical features and clinical presentations were typical of ELST. Sequencing studies of the tumor DNA disclosed a G to T substitution of nucleotide 564, which resulted in an amino acid substitution (Trp to Cys). This is the first report of the VHL gene mutation in a sporadic Japanese case of ELST. PMID- 11727271 TI - Alternative to histologic grading of cancer. PMID- 11727273 TI - The expanding world of molecular pathology. PMID- 11727275 TI - Statistical significance versus clinical relevance in cardiovascular medicine. AB - Evidence-based medicine is the foundation of everyday clinical practices and large clinical trials investigating the effects of various interventions on morbidity and survival and generally provide the most robust evidence. Cardiovascular medicine is considered one of the most evidence-based disciplines of medicine. However, there are a number of limitations to the general applicability of clinical trial results in cardiovascular medicine. Although generally useful to the clinician, clinical trials have often been suboptimally designed from 1 or several points of view. As a consequence of flaws in the design and the execution of the trials, statistical significance is quite often not equal to clinical relevance. This article outlines some of the shortcomings of designing and carrying out clinical trials, as well as inadequacies concerning the publication, interpretation, and implementation of the trial results. Evidence-based medicine is obviously not always as solid as one might think. PMID- 11727276 TI - The implantable cardioverter-defibrillator: does everybody need one? AB - The implantable cardioverter-defibrillator (ICD) has emerged as an effective, but expensive, therapy for arrhythmic sudden cardiac death. ICD use has been increasing by 20% to 30% per year. Clinical trials have shown that the ICD can be effective for both the primary prevention and the secondary prevention of sudden cardiac death in selected populations. Despite the available trial evidence, several issues pertaining to ICD use remain unresolved, including the treatment of patients not represented in clinical trials, the optimal selection of patients who will benefit from an ICD, the duration of benefit from an ICD, the quality of life for patients with an ICD, and both the cost-effectiveness and the cost impact of the ICD. These considerations are discussed in this article. PMID- 11727277 TI - Intravenous glycoprotein IIb/IIIa inhibition in non-ST segment elevation acute coronary syndromes. AB - Over recent years, substantial clinical trial evidence regarding glycoprotein IIb/IIIa inhibition for the medical management of non-ST segment elevation acute coronary syndromes has been compiled. Despite being recently advocated for the management of coronary instability within widely accepted guidelines, its use among patients presenting with acute coronary syndromes remains somewhat contentious. Within randomized placebo-controlled trials, uniform efficacy with the glycoprotein IIb/IIIa inhibitors has not been shown, whereas a disturbing excess in adverse events is evident within some trials. Currently, the basis for this heterogeneity of clinical evidence has not been adequately explained. However, evolving insights from clinical trials and basic research have further refined our understanding of glycoprotein IIb/IIIa antagonist therapy and the potential effects beyond the inhibition of the fibrinogen receptor. Likewise, appreciation of the pharmacokinetic characteristics of these agents provides putative explanations for the diverse findings of the randomized trials. Reexamination of the clinical trial data in light of this recent evidence provides a basis for interpreting the marginal results of glycoprotein IIb/IIIa inhibition in acute coronary syndromes. Consideration of these factors may facilitate the optimal clinical application of this class of agents to the management of coronary instability. PMID- 11727278 TI - Calcium channel blockers in treatment of hypertension. AB - Calcium channel blockers (CCBs) are among the most often prescribed drugs for the treatment of hypertension, but there is still uncertainty regarding the risks and benefits of their use as first-line drugs in the treatment of hypertension. Compared with placebo, dihydropyridine CCBs (long-acting nifedipine and nitrendipine) reduce the risk for cardiovascular endpoints, and in a pooled analysis of available studies on treatment of hypertension, significantly decrease the risk for strokes and cardiovascular and total mortality. This also holds true for patients with diabetes who have a clearly reduced risk when treated with CCBs as compared with placebo. However, compared with other active treatments in mixed study populations, CCBs are associated with a small risk increase for myocardial infarction and heart failure, but for cardiovascular mortality, there is only a very small and nonsignificant trend to a risk increase, and total mortality is similar. Among patients with diabetes, compared with angiotensin-converting enzyme inhibitors in particular, available data suggest that CCB use is associated with a moderate increase in cardiac endpoints. Therefore, among patients with diabetes and those with heart failure, angiotensin converting enzyme inhibitors are preferable as first-line drugs; among the large fraction of patients without these conditions, there is no convincing evidence that long-acting dihydropyridine or nondihydropyridine CCBs are inferior to other blood pressure-lowering drugs. In these patients, the choice of blood pressure lowering medication can be based on the expected tolerability, costs, and personal preferences. PMID- 11727279 TI - Coronary microembolization. AB - Atherosclerotic plaque rupture is a key event in the pathogenesis of acute coronary syndromes and during coronary interventions. Atherosclerotic plaque rupture does not always result in complete thrombotic occlusion of the entire epicardial coronary artery with subsequent acute myocardial infarction; however, in milder forms it may result in the embolization of atherosclerotic and thrombotic debris into the coronary microcirculation. The present report summarizes the available morphologic evidence for coronary microembolization in patients who died of coronary artery disease, especially sudden death. The report then goes on to address the experimental pathophysiology of coronary microembolization in animal models of acute coronary syndromes and heart failure. Finally, the report presents the available clinical evidence for coronary microembolization, highlights its key features--arrhythmias, contractile dysfunction, infarctlets and reduced coronary reserve--and addresses prevention by mechanical protection devices and glycoprotein IIb/IIIa antagonism. PMID- 11727280 TI - Fundamentals of the psoralen-based Helinx technology for inactivation of infectious pathogens and leukocytes in platelets and plasma. AB - Psoralens plus ultraviolet A (UVA) light inactivate viruses and bacteria as well as leukocytes. A system employing the synthetic psoralen compound amotosalen hydrochloride (S-59), in combination with UVA light, is being developed to decontaminate platelet concentrates and plasma in a blood-bank setting. S-59 is a heterocyclic psoralen compound that reacts by a three-step process with nucleic acids (NAs): (1) S-59 intercalates into the double helix; (2) upon illumination with long-wavelength ultraviolet light (UVA), it covalently attaches to a single strand, forming a monoadduct; and (3) additional illumination causes a photoreaction of the monoadduct with the second NA strand, resulting in an interstrand crosslink. The reaction occurs with the genomic material of DNA- and RNA-based viruses and occurs in genomes that are single stranded as well as double stranded. Inactivation rate is related to genome size. Large genomes such as those in leukocytes are far more susceptible to inactivation than are viruses such as hepatitis B virus (HBV), which is inactivated (>10(5) logs) under conditions being developed for blood-bank use. The efficiency of the process is affected by a number of practical considerations such as solution components and light source. The S-59 photochemical treatment process (PCT) has been optimized for platelet concentrates as currently processed for transfusion. PMID- 11727281 TI - Preclinical safety of a nucleic acid-targeted Helinx compound: a clinical perspective. AB - Helinx technology (Cerus Corp, Concord, CA) uses amotosalen HCl (S-59) and ultraviolet A (UVA) light in an ex vivo photochemical treatment (PCT) to inactivate viruses, bacteria, and leukocytes in platelet concentrates while preserving therapeutic function. A comprehensive preclinical safety program was conducted, which included carcinogenicity, single-dose and multiple-dose (up to 13 weeks' duration) toxicity, safety pharmacology (central nervous system [CNS], renal, and cardiovascular), reproductive toxicity, genotoxicity, vein irritation, phototoxicity, and toxicokinetic testing. The results of the toxicokinetic analyses indicated that the test articles provided large multiples of the clinical exposure to S-59, whether the comparison was based on dose, maximum plasma concentration, or area under the concentration-time curve. No specific target organ toxicity, reproductive toxicity, or carcinogenicity was observed. S 59 and/or PCT formulations demonstrated CNS toxicity, electrocardiographic (ECG) effects, and phototoxicity at supraclinical doses. On the basis of the extremely large safety margins, the CNS and ECG observations (at >30,000-fold the expected clinical exposure) as well as the results of genotoxicity and phototoxicity studies are not considered to be of toxicological relevance. The results of an extensive series of studies have thus demonstrated no toxicologically relevant effects of platelets treated with Helinx technology. PMID- 11727282 TI - Bacterial contamination of platelet concentrates: incidence, significance, and prevention. AB - Severe transfusion reactions associated with bacteria and/or their products, during or following a blood transfusion, were one of the earliest recognized complications of allogeneic blood transfusions. Bacterial contamination of blood products has thus been a problem for many decades and at present is likely the most common microbiological cause of transfusion-associated morbidity and mortality. Transfusion-associated sepsis due to contaminated platelet concentrates appears to be much more common than that due to contaminated red blood cells. The overall incidence of contaminated cellular blood products is approximately 1 in 3,000. However, transfusion to a recipient of a contaminated platelet unit may not necessarily be associated with clinically apparent morbidity, because the majority of contaminated platelet units contain relatively few organisms. In a minority of instances, contaminated units contain large numbers of potentially virulent bacteria, as well as endotoxins, and their transfusion is often associated with significant recipient morbidity and mortality. The incidence of severe septic episodes has not been clearly established, but is probably of the order of 1 per 50,000 platelet units transfused. With heightened awareness in recent years of the possibility that platelet transfusion-associated septic episodes can occur, a variety of measures have been proposed, and in some cases implemented, to try to prevent and control this transfusion risk. PMID- 11727283 TI - Inactivation of cytomegalovirus in platelet concentrates using Helinx technology. AB - Cytomegalovirus (CMV) infection in immunocompromised patients may result in severe morbidity and mortality. The standard of care for these patients is the use of CMV-seronegative blood components. Alternatively, filtered, leukocyte reduced blood components have been used. However, neither approach completely prevented transfusion-associated CMV infection in the recipients. The Helinx technology (Cerus Corp, Concord, CA) utilizing amotosalen hydrochloride (HCl) (S 59) and long-wavelength ultraviolet A (UVA) light, was developed to enhance the safety of platelet transfusion. The combination of 150 micromol/L of S-59 and a 3 J/cm2 UVA treatment inactivated greater than 10(5.9 +/- 0.3) plaque-forming units (pfus) per milliliter of CMV in full-sized therapeutic platelet concentrates. The efficacy margin of this treatment is greater than 100-fold. In an immunocompromised in vivo murine transfusion model, mice transfused with platelets contaminated with murine CMV (MCMV)-infected splenocytes became MCMV positive, exhibited histologic evidence of CMV disease, and died. Mice transfused with Helinx-treated platelets contaminated with MCMV-infected splenocytes prior to treatment remained MCMV-negative with no histologic evidence of CMV disease and remained healthy. These results demonstrated that Helinx treatment of MCMV contaminated platelet concentrates prevented MCMV infection by platelet transfusion in immunodeficient animals. In conclusion, Helinx technology offers a potential alternative to selection of CMV-seronegative units and to leukocyte reduction filtration as a means to decrease the risk of transfusion-acquired CMV infection. PMID- 11727285 TI - Design of clinical trials to evaluate the efficacy of platelet transfusion: the euroSPRITE trial for components treated with Helinx technology. AB - Methods of collection, preparation, and transfusion of platelet components have evolved markedly since the introduction of modern platelet transfusion therapy three decades ago. Despite these improvements, few randomized, prospective, controlled studies have been conducted to evaluate the effects of these innovations on the outcome of platelet transfusion--prevention and treatment of bleeding due to thrombocytopenia. The majority of studies have used posttransfusion platelet count increments (CIs) as the primary outcome variable rather than bleeding assessments. In general, these studies have only examined average values for platelet CIs or adjusted ratio measures such as corrected count increment (CCI). Because platelet transfusions are given repeatedly over variable periods of time, this type of analysis has not provided information about the effects of multiple platelet transfusions or about specific product or patient-related covariates that may impact the outcome. Longitudinal regression analysis of platelet CIs offers the potential to provide more information than simple average values of ratio measures. The euroSPRITE trial, a European, multicenter, phase III study undertaken to assess the clinical efficacy and safety of platelets prepared with Helinx technology (Cerus Corp, Concord, CA), used longitudinal regression analysis to characterize more fully the response to platelet transfusions with products prepared with this new pathogen inactivation technology. In contrast to previous studies, the euroSPRITE study examined peritransfusion hemostasis and global indices of hemostasis to correlate the effect of platelet CI with prevention and treatment of bleeding during a period of platelet transfusion support. PMID- 11727284 TI - Prevention of transfusion-associated graft-versus-host disease by inactivation of T cells in platelet components. AB - Patients with hematological malignancies and infants with congenital immunodeficiencies who received blood are two of many populations at risk for transfusion-associated graft-versus-host disease (TA-GVHD). Of the methodologies (eg, photoinactivation, peglyation, ultraviolet light, and irradiation) that can be used to prevent TA-GVHD, only irradiation of whole blood and cellular components is currently accepted practice of the US Food and Drug Administration (FDA). Among the newer methods that have been developed to reduce the risks of bacterial and viral contaminants of platelet transfusions, photochemical treatment (PCT) using psoralens and long-wavelength ultraviolet (UVA) irradiation modifies bacterial and viral genomes sufficiently to inhibit replication. Among a broad group of compounds, the synthetic psoralen compound amotosalen hydrochloride (HCl) (S-59) has been shown to be particularly effective in inactivating bacteria and viruses, without adversely affecting in vitro and in vivo platelet function. PMID- 11727286 TI - The doctor on the patient's turf: assessing patient satisfaction with physician home visit programs. AB - We evaluated a physician home visit program (n = 23 patients) focusing on program implementation and quality. Quality was measured by evaluating patient satisfaction with services using a patient satisfaction scale and interviews with patients, caregivers, and providers. Scale results showed patients expressed the highest satisfaction with access to routine care and physician consideration. Patients expressed less satisfaction with access to emergency care and continuity of care. Physician communication and integration with home- and community-based service providers were other areas of concern. Recommendations include enhancing physician communication skills in the home, providing care for urgent medical conditions, improving chart documentation, and incorporating community-based chronic care experts into the program. PMID- 11727287 TI - Consumer choice in home- and community-based long-term care: policy implications for decisionally incapacitated consumers. AB - There is a strong national and international movement to enhance the role of consumer choice, control, and direction regarding important aspects of publicly financed home- and community-based long-term care. The research project described in this article set out to examine, with a particular focus on Ohio, policy relevant issues pertaining to how choices about publicly financed home- and community-based long-term care are actually made, implemented, and monitored under a consumer direction model, but when the care recipient is unable to act as an autonomous, independent consumer selecting rationally among various home- and community-based long-term care options competing for the consumer's business in the marketplace. PMID- 11727288 TI - Improving dementia care through community linkages: a multi-site demonstration project. AB - The purpose of the multi-site project was to develop and implement a model for dementia care which improved linkages of caregivers to community services. Key components of the model included a single point of informational contact, provider education, case-finding, caregiver education and support, internal linkages, and linkages with community services. The model was implemented at six medical centers. Outcome measures included caregiver, provider, and community agency satisfaction. Caregivers reported high satisfaction with information provided to them about community resources. Primary care providers reported that dementia services had improved from one year earlier. Community agencies reported high satisfaction with the dementia program initiatives. PMID- 11727289 TI - Reflection in action in caring for the dying: applying organizational learning theory to improve communications in terminal care. AB - Currently, single loop learning is the predominant method of problem solving orientation engaged in by healthcare institutions. This mode of learning is not conductive to fostering needed communications between health care providers and terminal patients. Reflection in action, second loop learning, focuses on deep listening and dialogue and can be critical in opening communications paths between the dying patient and his or her caregivers. This article discusses organizational learning theory and applies the theories double loop learning technique of reflection in action to end-of-life care. The article further explores an exemplar of reflection in action in a Palliative Care Program, and end-of-life home care program at Kaiser Permanente. In order to more effectively meet the needs of terminally ill patients, greater efforts are needed to incorporate second loop learning into the practice of those caring for these patients. PMID- 11727290 TI - An improved scale for assessing patients' trust in their physician. AB - Patients' trust in their primary care physician is a critical concept for healthcare practitioners and scholars. At the clinical level, such trust buttresses patient-physician treatment relationships; at the organizational level, such trust fosters enhanced organizational effectiveness and other positive outcomes. To empirically assess various trust-related issues on both levels, we develop a comprehensive, bi-dimensional trust scale specific to patient-physician relationships. Response analysis from two samples suggests that the scale's benevolence dimension comprises understanding patients' individual experiences, expressing caring, communicating clearly and completely, building partnership and sharing power, demonstrating honesty and respect, and keeping information confidential. The scale's technical competence dimension comprises evaluating problems thoroughly, providing appropriate and effective treatment, predisposing factors, and structural and staffing factors. PMID- 11727291 TI - Patients' perceptions of service quality dimensions: an empirical examination of health care in New Zealand. AB - The 1984 liberalization of the New Zealand economy has resulted in a health care sector that has become very competitive (Zwier and Clarke, 1999). The private sector is now able to supply health care services and, as a result, a greater value is being placed on patient satisfaction (Zwier and Clarke, 1999). However, despite the increasing focus on customer satisfaction, research into health care patients' perceptions of the dimensions of service quality is scarce. This can be problematic, as quality of care is an essential issue in the strategic marketing of health care services (Turner and Pol, 1995). This study takes a step towards addressing this deficiency by identifying patients' perceptions of the dimensions of service quality in health care. The findings of this study are based on the empirical analysis of a sample of 389 respondents interviewed by telephone. The findings indicate that the service quality dimensions identified in this health care specific study differ in number and dimensional structure from the widely adopted service quality dimensions first identified by Parasuraman, Berry and Zeithaml (1988): reliability, responsiveness, assurance, empathy and tangibles. The service quality dimensions identified in this study were: reliability, tangibles, assurance, empathy, food, access, outcome, admission, discharge and responsiveness. In addition, health care patients perceive the service quality dimensions relating to the core product in health care delivery (for example, outcome and reliability) as more important than the service quality dimensions relating to the peripheral product in health care delivery (for example, food, access and tangibles). Finally, the results of this study suggest that patients with different geographic, demographic, and behavioristic characteristics have different needs and wants during health care delivery and therefore perceive different service quality dimensions as important. PMID- 11727292 TI - Strategic decision-making among top executives in acute-care hospitals. AB - Recent changes within the dynamic healthcare industry have necessitated further assessment of the strategic thought processes associated with financial success. Therefore, this study explored hospital executive's use of a comprehensive strategy-making process and its impact on performance in acute care hospitals. Taken as a whole, the strategy-making process is found to be positively associated with financial performance. Supplementary analysis, investigating the unique effects of environmental planning, scanning and analysis, revealed that, when considered separately, scanning and analysis contribute significantly to performance, whereas strategic planning, as classically defined, does not. Implications for decision-making in hospital settings are discussed. PMID- 11727293 TI - Consumer involvement: effects on information processing from over-the-counter medication labels. AB - The objective of this study was to evaluate the effects of consumer involvement on information processing from over-the-counter (OTC) medication labels. A sample of 256 students evaluated simulated OTC product labels for two product categories (headache and cold) in random order. Each participant evaluated labels after reading a scenario to simulate high and low involvement respectively. A questionnaire was used to collect data on variables such as label comprehension, attitude-towards-product label, product evaluation, and purchase intention. The results indicate that when consumers are involved in their purchase of OTC medications they are significantly more likely to understand information from the label and evaluate it accordingly. However, involvement does not affect attitude towards-product label nor does it enhance purchase intention. PMID- 11727294 TI - The new social marketing challenge to promote radon testing. AB - As part of a project funded by the Environmental Protection Agency, exploratory qualitative analysis was conducted to gain insight into perceptions of the threat of radon in the Karst geological region (i.e., Northern Alabama, Central Tennessee, Central Kentucky). Based on health practitioner input, it was clear that the tenets of Protection Motivation Theory (PMT) and the probing afforded by focus group research would provide greatly needed theory-based insight into the public reactions (or lack thereof) to the threat posed by radon. Qualitative research findings of this project are discussed as well as preliminary recommendations are provided to advance the protection motivation theory research agenda for promoting awareness of the threat of radon and to influence appropriate response to that threat. PMID- 11727295 TI - Physicians' perceptions and uses of commercial drug information sources: an examination of pharmaceutical marketing to physicians. AB - Data were collected from physicians attending a medical conference. This exploratory study was primarily interested in two areas. First, the investigators were interested in better understanding physicians' responses to different promotional tactics typically used by the pharmaceutical industry. Pharmaceutical representatives were most useful, followed by drug samples and infomercials in medical journals. Direct mail, promotional faxes, and promotional products were used less by physicians. Second, the investigators were interested in learning what information sources influenced physicians' drug choices. Physicians were primarily influenced by their prior experience with a drug, then by drug compendiums, and journal articles. Physicians were also influenced by information provided by the industry and other factors, like the drug's price and their patients' financial situations. Managerial implications for marketing to physicians and ideas for future research are discussed. PMID- 11727296 TI - [How a formula should be presented in a book]. PMID- 11727297 TI - [Malignant melanoma and its treatment]. PMID- 11727298 TI - [The concept of sentinel lymph nodes. Principles and clinical significance]. PMID- 11727299 TI - [Allergic rhinitis. Is allergen therapy a possible treatment?]. PMID- 11727300 TI - [Myocardial infarct and acute coronary syndrome. New research results or acute treatment and secondary prevention]. PMID- 11727301 TI - [Contraceptives for epileptic?]. PMID- 11727302 TI - Web alert. The chemistry of drug design and lead optimization. PMID- 11727303 TI - Growth factor receptor kinase inhibitors: recent progress and clinical impact. AB - Inhibition of growth factor receptor kinases is one of the most promising therapeutic approaches for the treatment of cancer. This review focuses on the most recent progress in this area, and gives an overview of the compounds currently in the clinic, as well as key preclinical analogs. PMID- 11727304 TI - Progress towards testing the amyloid hypothesis: inhibitors of APP processing. AB - Alzheimer's disease is the most common form of dementia and a major public health problem. The amyloid hypothesis suggests that Alzheimer's disease is due to the abnormal accumulation of amyloid-beta protein (A beta) in affected brain regions. Rational therapies aimed at reducing amyloid burden in brain are currently being pursued in preclinical and early clinical development. This review summarizes recent progress in understanding the beta- and gamma-secretase activities required for the formation of A beta peptide and discusses therapeutic strategies aimed at inhibiting these activities. Recent progress in the identification of small molecule inhibitors of these secretases is also reviewed. PMID- 11727305 TI - Inhibition of HIV-1 integrase by small molecules: the potential for a new class of AIDS chemotherapeutics. AB - HIV-1 integrase, one of three constitutive viral enzymes required for replication, is an attractive target for chemotherapeutic intervention in the treatment of AIDS. Unlike the retroviral reverse transcriptase and protease enzymes, successful drug candidates based on the inhibition of integrase have yet to emerge despite the multitude of laboratories working on the problem. In vitro inhibition of integrase in the laboratory has not always led to antiviral activity in cell culture. Indeed, most compounds found from broad screening efforts have fallen into this category. Only recently have compounds that inhibit enzymatic function as well as exhibiting an antiviral effect as a result been discovered. The reasons why certain structural classes of compounds fail to have antiviral activity are only beginning to be understood. In the last two years a new class of compounds has emerged that do produce potent inhibition of viral growth in cell culture. Further work in this area may yet produce clinically useful antiviral agents. PMID- 11727306 TI - Recent advances in the discovery of small molecule therapies for HCV. AB - Hepatitis C virus (HCV) is the leading cause of chronic hepatitis in humans. This virus, first identified in the late-1980s, has been the target of aggressive efforts by the pharmaceutical industry to identify novel therapeutic agents. Current therapy requires the use of interferon-alpha in conjunction with ribavirin. Regrettably, the utility of this therapy is limited by both an unsuitable side effect profile and low efficacy. These shortcomings have maintained the pressure to identify more effective therapies. Several compounds that take advantage of different strategies for achieving an antiviral effect are currently in clinical trials. Several more classes of novel antiviral agents have been identified using a range of in vitro assays. The recently developed stable cell-based assay for HCV will be a highly useful tool in on-going discovery efforts. PMID- 11727307 TI - Thrombin receptor antagonists as novel therapeutic targets. AB - In addition to its pivotal role in hemostasis, thrombin activates various cell types such as platelets and vascular smooth muscle cells via proteolytic processing of specific cell-surface receptors known as proteinase activated receptors (PARs), the prototype of which is PAR-1. Thrombin receptor activation is likely to play a key role in cardiovascular disorders such as arterial thrombosis, atherosclerosis and restenosis, and as such a thrombin receptor antagonist should have potential utility in the treatment of these disorders. Since the fibrin pathway is unaffected by thrombin receptor antagonism, a thrombin receptor antagonist is expected to have minimal bleeding liability, which is a complicating factor in existing antithrombotic therapy. The currently available collection of thrombin receptor antagonists fall into three categories: (i) peptide antagonists; (ii) peptidomimetics; and (iii) non-peptide thrombin receptor antagonists, and this review outlines the development of members of these classes. PMID- 11727308 TI - High-throughput organic synthesis of peptide mimetics. AB - This article is a subjective review of the literature from 1999 to March 2001. It seeks to highlight current progress in the high-throughput synthesis of peptide mimetics. The review is loosely classified into sections based on the type of secondary structural feature which is being mimicked (reverse turn, beta-strand, peptide surrogates) or a common motif. The examples chosen have demonstrated the ability to produce a large number of compounds rapidly or, in our opinion, the method could produce large numbers of compounds if desired. PMID- 11727309 TI - Total synthesis of natural product DNA cross-linking agents. AB - A variety of naturally occurring compounds interact with duplex DNA by the formation of covalent cross-links. This review describes recent progress on the total synthesis of four natural product families, including the mitomycins, the structurally and functionally related agent FR-900482, the bis-pyrrole isochrysohermidine and the azinomycins. PMID- 11727310 TI - Combinatorial lead discovery and optimization of antimicrobial oxazolidinones. AB - Oxazolidinones are a new class of synthetic protein synthesis inhibitors with a novel mode of action, which are active against multidrug-resistant pathogens. The recent approval of the first drug of this class linezolid (Zyvox, Pharmacia) has sparked further interest in the development of novel antibiotics of this structural type. Combinatorial chemistry is an important tool capable of facilitating the discovery and optimization of the next generation leads with enhanced potency and improved activity spectrum. Combichem integration into a medicinal chemistry project is a multifactorial task that is reviewed herein in a context of efforts targeting novel oxazolidinone antimicrobials. Key strategic considerations that govern projects of this type are analyzed, and a survey of combichem methodologies and oxazolidinone libraries is presented with attention to biological evaluation results. PMID- 11727311 TI - Combinatorial synthesis of aminoglycoside libraries. AB - The interaction of aminoglycosides with RNA represents a paradigm in the use of small molecules as effectors of RNA function. Aminoglycosides bind and affect the function of a variety of therapeutically useful RNA targets and show antimicrobial as well as antiviral activities. However, due to the complex nature of aminoglycosides, efforts for synthesizing large libraries of these structures to screen against prospective biological targets have been limited. This review covers the recent advances in the generation of aminoglycoside libraries. PMID- 11727312 TI - The use of bioisosteric groups in lead optimization. AB - It is now half a century since Friedman introduced the term bioisosterism for the similar biological activity of structurally related compounds. Since then, the concept has been used extensively and successfully in the optimization of lead compounds in drug discovery. The number of chemical lead compounds has expanded enormously in recent years due to the expression of an increasing number of recombinant proteins, and the screening of these new protein targets against a large number of compounds in high-throughput screens. For the fine-tuning of lead compounds to obtain candidates suitable for clinical trials, which is in most circumstances still a tedious process, the use of bioisosteric replacement can be of significant value. This is especially the case in optimizing for selectivity for a specific target and in improving the pharmacokinetic properties of lead compounds. The use of bioisosteres in lead optimization is illustrated by some recent examples from the literature. PMID- 11727313 TI - NMR in the acceleration of drug discovery. AB - The field of NMR spectroscopy has grown beyond expectations from the first historic observation of nuclear magnetic resonance in 1946 by Bloch and Purcell, to the first generation of protein structures 20 years ago, to the present where NMR structures now represent 15 to 20% of those submitted to the Protein Data Bank (PDB). However, the applications of NMR go far beyond structure determination. Since NMR is a non-invasive technique that provides a wealth of information about structure and dynamics, it is an ideal analytical method that is now being used in every step of the drug discovery and development process. Indeed, NMR is undergoing a renaissance in the pharmaceutical industry as a plethora of new applications have been developed, ranging from in vitro screening methods in early stage discovery to in vivo magnetic resonance imaging techniques used to assess drug efficacy in clinical trials. In this review, we will present an overview of the role of NMR in these areas, with a special focus on how NMR is poised to revolutionize the high-throughput structural characterization of protein-ligand interactions. PMID- 11727314 TI - Combinatorial approaches to formulation development. AB - This review describes the use of combinatorial methods for the development of drug formulations. Combinatorial methods are applied to find solutions to various formulation problems, including drug solubilization, controlled release, oral drug administration, and others. Various methods are described, including the synthesis of carrier libraries, high-throughput screening and computational analysis, which are used during the formulation development process, starting from initial assays through the optimization of formulation composition, to the optimization of the manufacturing process. This review also describes an integrated approach to drug formulation development using libraries of block copolymers as the drug carriers. PMID- 11727315 TI - Computational methods to estimate drug development parameters. AB - Computational methods are currently available to estimate oral bioavailability, solubility, metabolism, toxicity, pKa, blood-brain barrier permeability and other ADME and physicochemical parameters. Decisions as to which methods to implement and to employ must be made in accordance with the stated goals of a drug discovery organization, the timeline for these goals, and the budgetary limitations as set forth to accomplish these goals. Certain methods are more attractive to the production environment of a pharmaceutical project team where early ADME and Tox information is sought to aid in drug design decisions and prioritization. Practical limitations of these methods, ease of use, utility of results, as well as their scope and limitations are discussed. Recommendations as to which parameters are best estimated by commercial products, as opposed to those that can be developed in-house, are delineated. Special attention is given to those methods that can be integrated into the current high-throughput paradigms of drug discovery programs. Together, these considerations define a 'zero-infrastructure' approach to provide ADME and Tox information during the early stages of the drug design process. PMID- 11727316 TI - Computational methods to predict drug safety liabilities. AB - Computational methods to predict drug safety liabilities are reviewed. A special emphasis of this article is on the perceived strengths and weaknesses of the commercial turnkey predictive toxicology programs (TOPKAT, MULTICASE and DEREK). This article includes proposals for improvements of individual predictive programs, experiences with pharmaceutical datasets, evaluations incorporating multiple programs, and strategies of their use as a sentinel filter for liability assessment early in the drug discovery process. PMID- 11727317 TI - Advances in protein microarray technology for protein expression and interaction profiling. AB - Protein microarrays address a great demand for high-throughput protein analysis techniques. Because protein microarrays detect many proteins in parallel, are quantitative, and have minimal reagent and sample consumption requirements due to miniaturization, they are potentially powerful tools for applications in basic and applied biology. Advances in manufacturing, protein immobilization and detection methods have enabled high-throughput protein quantitation and interaction studies. Protein microarrays can be applied to protein function studies, screening the production of antibodies and recombinant proteins, discovery of proteins implicated in disease or that are potential drug targets, and rapid detection or diagnosis of disease. A remaining challenge for the full implementation of protein microarrays in the acquisition of large sets of high affinity and highly specific protein capture reagents. PMID- 11727318 TI - Assay technologies for screening ion channel targets. AB - The sequence of the human genome will soon provide researchers with hundreds of new ion channel genes. To create a successful ion channel drug discovery program, it is necessary to quickly develop reliable and robust high-throughput screening (HTS) assays for those ion channels implicated in important diseases. Ion channels are dynamic proteins, and therefore require assays that 'sense' their various functional states. Competition-binding assays, although successfully used for other target classes, often fail to identify ligands that modulate specific ion channel functions. Cell-based functional assays, therefore, are preferred for HTS of ion channel targets. In this review, we evaluate the various cell-based assays available for ion channels, and discuss future directions for assay improvements. PMID- 11727319 TI - Web alert. The chemistry of metabolic and toxicological processes. PMID- 11727320 TI - In vitro carcinogenicity testing: present and future perspectives in pharmaceutical development. AB - Almost all new drugs must be tested for carcinogenicity at some point during their development, and ultimately, a lifetime in vivo assay, usually in rodents, must be performed. Many in vitro assays of carcinogenicity have been developed for use before short- or long-term in vivo testing in order to remove from the development stream those drugs that are likely to produce tumors in vivo. This review discusses in vitro assays that are required by the International Conference on Harmonization, followed by a discussion of in vitro carcinogenicity assays, which are currently in use, but are not specifically required. The concluding section is devoted to a discussion of high-throughput compatible carcinogenicity screens and potential human cell-based high-throughput compatible screens with reference to future methods in silico. PMID- 11727321 TI - The prediction of human clearance from hepatic microsomal metabolism data. AB - Human liver microsomal intrinsic clearance has become a commonly measured parameter during drug discovery, and such data are used to design compounds predicted to possess optimal drug disposition characteristics. Liver microsomal intrinsic clearance values can be scaled and used to predict hepatic clearance in humans. Clearance, when combined with the volume of distribution, determines the half-life of a drug. Hepatic clearance, when combined with absorption, determines the oral bioavailability of a drug. Half-life and oral bioavailability are key determinants of the dosing regimen, i.e., size of dose and frequency of administration. Thus, the accurate prediction of human clearance is important in the selection of new compounds for progression into development, as new drugs on the market must not only be efficacious and safe, but must also be convenient to use for patients and physicians. Over the past decade, exploring methods whereby human clearance can be predicted from in vitro data has been an area of active research in drug metabolism science. Human liver microsomes have been a key tool in this research. This in vitro system possesses many of the major drug metabolizing enzymes and is thus applicable to a wide variety of compounds. This review describes the theoretical and practical aspects of predicting clearance from human liver microsomal intrinsic clearance data, a summary of advantages and shortcomings of this in vitro system, a synopsis of recent applications of human liver microsomal intrinsic clearance data in clearance predictions, and a discussion of potential future directions for this field. PMID- 11727322 TI - The potential pharmacological and toxicological impact of P450 screening. AB - Cytochrome P450 (CYP) proteins catalyze the majority of drug biotransformations and play a pivotal role in several homeostatic mechanisms. Perturbation of CYP activity can have profound effects on therapeutic efficacy and in extreme cases may lead to life-threatening toxicities. The trend towards front loading physicochemical properties, and drug metabolism and pharmacokinetics (DMPK), in an attempt to minimize the attrition within drug development, has been a concentrated effort on screens aimed at optimizing interactions with the CYP system. These include improving metabolic stability and minimizing the potential for drug-drug interactions associated with marked reductions (inhibition) or increases (induction) in CYP activity. Increased emphasis on the identification of individual CYPs responsible for drug metabolism is also a common theme, with the desire to provide early alerts to compounds cleared predominantly by polymorphic enzymes or isozymes which demonstrate marked inter-subject variability. These data may provide a rationale for individualized therapy in the near future. Such screens should see the establishment of large databases and better in silico (if not in cerebro) prediction of properties typically associated with successful marketed therapies. PMID- 11727323 TI - Prediction of a new drug's potential to cause idiosyncratic reactions. AB - It is currently impossible to accurately predict which new drugs will be associated with a significant incidence of idiosyncratic drug reactions and this introduces a significant degree of uncertainty into the drug development process. In the absence of a better understanding of the mechanisms of these reactions, there are a few screening procedures that would likely reduce the probability that a new drug will be associated with idiosyncratic drug reactions. One method is to screen candidates for the formation of reactive metabolites and halt development of drugs that form significant amounts of such metabolites. However, such metabolites are not easy to screen for, and it would also eliminate many candidates that would have been safe if developed. Another risk factor may be the ability of the reactive metabolite to cause cell damage. Even though idiosyncratic reactions appear to be immune-mediated, reactive metabolites that cause cell damage will likely increase the probability of an immune response. Simply developing more potent drugs is likely to decrease the incidence of idiosyncratic drug reactions and it appears that drugs given at a dose of 10 mg/day or less, are associated with a low incidence of idiosyncratic drug reactions. Although the general use of these methods would probably lead to safer drugs, they are far from satisfactory. The use of genomics to search for patterns of change in gene expression that are associated with drugs that cause idiosyncratic reactions has the potential to lead to a more effective screen, but this is unlikely to be a simple process. Ultimately, it is likely that a much better understanding of the mechanisms involved in such reactions will be required to make real progress. PMID- 11727324 TI - Prediction of drug allergenicity: possible use of the local lymph node assay. AB - The mouse local lymph node assay (LLNA) is currently used for the prospective identification of chemicals that have the potential to cause skin sensitization and allergic contact dermatitis. In this respect, the assay has been fully and formally evaluated and validated, and has been accepted recently as a stand-alone method for the identification of potential skin sensitizers. The assay involves topical application of test substance and the subsequent measurement of proliferative responses in the lymph nodes draining the site of exposure. The testing of new drug entities using a similar assay technique could offer a potential alternative for the identification of potential drug allergens. Currently, the popliteal lymph node assay (PLNA), or modifications of it, are used in research studies for the identification of drugs which have the potential to cause allergic or autoimmunogenic reactions. The PLNA involves subcutaneous application of test substance into the hind footpad, followed by the measurement of proliferative responses, or other parameters of immune activation, in the popliteal lymph nodes. However, these assays have not been validated systematically and the potential utility of a modified LLNA for use in the identification of such compounds is discussed. PMID- 11727325 TI - Integrating metabolism and toxicity in multi-organ systems. AB - A wide range of experimental and predictive approaches are currently employed by the pharmaceutical industry in an attempt to avoid developing molecules with likely toxicities. Experimental systems may be adapted to handle the increasing number of molecules that require screening. The cost implication of such extensive screening is, however, immense and in silico models must therefore become the first lines of investigation to assess the metabolic and toxicological potential of new compound libraries. In silico systems have been based on the relationship between the chemical structure of novel molecules and toxicological outcomes in terms of types, but not always, locations of lesions. There should be sufficient published and in-house data available on the distribution of diverse molecules to identify the rules that govern organ, cellular and subcellular distribution of molecules (and their metabolites) as the basis of tissue and cell targeting and, more importantly, the possibility of toxicological processes. In addition, the extensive cell biology knowledge needs to be linked to current in silico systems to enhance our understanding of metabolic activation and other processes that affect likely toxicological consequences in different tissues and specific cell types. There is a need to provide a single knowledge platform that can access all current in silico products that use different datasets. The closer integration of these diverse tools will help predict other approaches to help identify the most likely toxicological potential of novel molecules, and ensure the 'least' toxic molecules are developed. PMID- 11727326 TI - Enhancing oral absorption in animals. AB - Optimizing absorption can present unique demands in many animal studies because where dose-response linearity and high tissue exposure levels are required, doses may be greater than when administered for therapeutic effect. At the same time, investigative molecular constructs designed to enhance potency and specificity can have features that militate against efficient absorption. Pre-treatment or formulation to optimize absorption may be warranted. This article reviews various physicochemical approaches employed for absorption enhancement. Strategies based on capitalizing or neutralizing physiological processes are also discussed, as there is much current interest in exploiting novel non-invasive routes to deliver medicinal agents. PMID- 11727327 TI - Standardized electroretinography in primates: a non-invasive preclinical tool for predicting ocular side effects in humans. AB - This paper reviews the use of a standard protocol for electroretinography in drug safety evaluation studies with non-human primates. The architecture and physiology of the eye of humans and higher primates are very similar, thus making the non-human primate a preferred animal model for evaluating potential ocular side effects of drugs in humans. Using the technical and procedural features of the human protocol, i.e., the standard for clinical electroretinography (SCE), electroretinography can be routinely performed from early- to late-phases of drug development as a toxicodynamic endpoint for retinal toxicity and potential neurotoxicity in non-human primates. Hence, scientists can collect clinically relevant data using a standardized testing procedure found in the SCE. This is important in drug safety evaluations since the SCE is an established protocol that lends itself easily for incorporation into studies requiring adherence to international Good Laboratory Practice regulations. The technique is non invasive, painless, and confers no adverse effects or damage to the eye. PMID- 11727328 TI - Chem-tox informatics: data mining using a medicinal chemistry building block approach. AB - Relating chemical structure to biological activity is not a new endeavor, however, the ability to do this on large datasets is just emerging. To cope with the enormous amounts of data being generated, an assortment of computational methods has been developed in the fields of chemoinformatics and computational toxicology. Many of the molecular descriptors used in these approaches are abstract, theoretical constructs that are difficult to understand and visualize. Having easily recognized chemical features, such as those in several new programs, will allow chemists to use toxicological information (or any biological information) when designing new libraries. These improved chem-tox informatics systems will have an impact on library design, hit and lead optimization, development candidate testing and regulatory review. PMID- 11727329 TI - Pathophysiologic characteristics of hypovolemic shock. AB - In the late 1800s, while caring for a trauma victim, Warren characterized shock as "a momentary pause in the act of death." A great deal about shock has been discovered since this first description. Dorland's Medical Dictionary defines shock as a condition of profound hemodynamic and metabolic disturbance characterized by failure of the circulatory system to maintain adequate perfusion of vital organs. Shock is now being defined at the cellular level as the inadequate delivery of nutrients to the cells of the body. Because oxygen is the only nutrient that cells cannot store in any appreciable quantity, shock is also equivalent to inadequate oxygen delivery (DO2). Although there are many types of shock, this article concentrates on the pathophysiologic characteristics of hypovolemic shock. PMID- 11727330 TI - Cardiogenic shock and cardiac arrest. AB - Severe heart disease may cause hypotension and hypoperfusion, and ultimately circulation may cease altogether. These two clinical syndromes are cardiogenic shock and cardiac arrest, respectively. This review summarizes the causes and clinical features of each, and describes the treatment options available to clinicians managing patients in cardiogenic shock. PMID- 11727331 TI - Systemic inflammatory response syndrome, sepsis, and multiple organ dysfunction. AB - Companion animals with sepsis and multiple organ dysfunction can be the most challenging of all patients to treat. Current research in humans and laboratory models offers some exciting insights into the pathophysiology behind some of our most frustrating clinical challenges. This article applies several current concepts to a clinical case of pancreatitis and secondary sepsis to illustrate some of the cardiovascular, immune, and coagulation abnormalities commonly seen. PMID- 11727332 TI - Central venous pressure and arterial blood pressure measurements. AB - Arterial blood pressure measurement and central venous pressure monitoring are important tools in the management of the critically ill pet. Central venous pressure is reflective of right atrial pressure and provides information concerning volume status. Arterial blood pressure is helpful in determining if perfusion to vital tissues is adequate. By providing more information with which to tailor fluid therapy and by prompt recognition of hypo- or hypertension, these monitoring tools are instrumental in the management of the critically ill pet. PMID- 11727333 TI - Cardiac output, wedge pressure, and oxygen delivery. AB - The primary goal of the intensive care clinician can be said to be to optimize global DO2. This approach is the primary means by which the greatest killers of the critically ill patient (sepsis, SIRS, multiple organ dysfunction syndrome) may be addressed at present. Optimizing DO2 means delivering just enough to meet the patient's needs, because therapeutic measures taken to increase DO2 are all associated with some degree of risk. When used correctly, the PAC can allow the clinician to determine if DO2 is optimal and, if not, what steps might be best suited to improve on it. Newer generations of PACs are becoming available and can provide valuable additional insights into a patient's cardiovascular status. Nearly all attempts to increase DO2 address one or more of a relatively short list of variables. The specific endpoints of therapy need to be tailored to the individual patient but include clinical, metabolic, organ function, and hemodynamic markers. As clinicians expand our understanding of the key elements found in survivors of critical illness, it is hoped that this knowledge translates into better outcomes. PMID- 11727334 TI - Colloid and crystalloid resuscitation. AB - Use of colloid and crystalloid fluids during resuscitation can be a complicated process. An understanding by the veterinarian of the patient's cardiovascular state, the underlying pathologic process, and the characteristics of crystalloid and colloid fluids available is necessary for establishing a fluid therapy plan. Through frequent reassessment and tailoring of the fluid plan according to the patient's response, the risks of fluid overload and fluid deficiency are reduced. PMID- 11727335 TI - Cardiovascular pharmacotherapy. Hemodynamic drugs and antiarrhythmic agents. AB - Successful management of the patient presenting with acute cardiovascular compromise requires a thorough working knowledge of the therapeutic agents available for treatment. This article focuses on the indications for, dosing, and monitoring employed for successful use of the vasoactive agents and antiarrhythmic medications in current widespread use in the veterinary intensive care setting. PMID- 11727336 TI - Cardiopulmonary cerebral resuscitation. AB - Cardiopulmonary arrest (CPA) is defined as the abrupt and unexpected cessation of spontaneous and effective ventilation and circulation. CPA can be the natural ending of a normal and long life; however, when CPA is the result of a reversible problem in an animal that has a treatable medical condition, rapid recognition and treatment may make the difference between a happy ending and premature death. Cardiopulmonary resuscitation provides artificial ventilation and circulation until advanced cardiac life support can be provided and spontaneous cardiopulmonary function is restored. The term cardiopulmonary cerebral resuscitation originated in the early 1960s in recognition of the severe central nervous system complications of prolonged cardiac arrest in human beings. Although neurologic complications of CPA may not be as noticeable in companion animals, newer brain-sparing strategies that recognize the consequences of reperfusion injury and the inflammatory cascade may some day offer improved survival. PMID- 11727337 TI - Strategies for management of acute renal failure. AB - Acute renal failure (ARF) is often defined as the sudden inability of the kidneys to regulate water and solute balance. ARF may be more broadly defined as rapid deterioration of renal function resulting in the accumulation of nitrogenous wastes such as urea and creatinine. Clinically, oliguria is defined as urine flow of less than 2 mL/kg/h and anuria has no measurable urine production. In animals, the most common cause of ARF is nephrotoxicity; ischemia ranks second, with interstitial and glomerular diseases following. PMID- 11727338 TI - Electrolyte disorders. AB - Abnormal electrolyte concentrations occur commonly in hospitalized patients and may produce a variety of clinical symptoms, cause lack of response to therapeutics for primary clinical conditions, and affect clinical outcome. Recognition of electrolyte disturbances requires a high index of suspicion by the clinician for such a disturbance and prompt therapy to ensure a positive and timely outcome for the patient. This article discusses electrolyte abnormalities that occur in critically ill patients, with a review of diseases commonly associated with each electrolyte disturbance, and their recommended management. PMID- 11727339 TI - Metabolic acid-base disorders. AB - Acid-base homeostasis is an important determinant of many physiologic functions. Nowhere is understanding the mechanisms and significance of hydrogen ion (H+) imbalance more important than in critical care management, where patients are threatened with a physiochemical disorder that is often as complex as it is dangerous. Although there may be contentious issues yet unresolved concerning acid-base homeostasis, the incontrovertible fact is that the body at least seems to defend H+ balance as vigorously as it does oxygen transport or perfusion pressure. Equally, there seems to be an important and predictable relation between this balance and other physiochemical variables such as concentrations of other ionic species, carbon dioxide, and plasma proteins. The prudent clinician strives to understand whether or not and how acid-base imbalances are affecting his or her patient and what to do about it. PMID- 11727340 TI - Respiratory acid-base disorders. AB - Respiratory acid-base disorders, although infrequently diagnosed in veterinary medicine, can cause or contribute to adverse clinical outcomes. Recognition of the mechanisms and causes of respiratory acidosis and alkalosis can prompt clinical detection of the acid-base derangement, allowing for appropriate intervention. PMID- 11727341 TI - The skin in old age. PMID- 11727342 TI - A back-pain patient seen moving furniture. PMID- 11727343 TI - Key developments in dermatology. PMID- 11727344 TI - Managing the patient with psoriasis. PMID- 11727345 TI - Identifying disease in pigmented skin. PMID- 11727346 TI - Casebook: the itchy patient. PMID- 11727347 TI - Erythema multiforme and erythema nodosum. PMID- 11727348 TI - The mouth in dermatological disorders. PMID- 11727349 TI - Cryotherapy in general practice. PMID- 11727350 TI - Rational prescribing of statins. PMID- 11727351 TI - Inner secrets of paper 1. PMID- 11727352 TI - Sunshine is therapy, too. PMID- 11727353 TI - Kallmann's syndrome: pregnancy through intracytoplasmic sperm injection and complicated by gestational diabetes. AB - Kallmann's syndrome is characterized by hypogonadotropic hypogonadism and anosmia. Assisted reproductive techniques such as intracytoplasmic sperm injection (ICSI) may be required to treat the infertile couple with oligozoospermia. Rare complications have been described in patients with Kallmann's syndrome, but gestational diabetes has not previously been reported. A case of Kallmann's syndrome with infertility is reported. Ovulation was successfully induced by human menopausal gonadotropin therapy, but pregnancy could not be achieved by artificial insemination or by conventional in vitro fertilization, although the husband had only moderate oligozoospermia. A high fertilization rate of the retrieved oocytes and successful pregnancy was achieved by ICSI. The pregnancy was complicated by gestational diabetes that was managed by insulin therapy. Successful ovulation induction in Kallmann's syndrome is not rare, but ICSI may be needed in selected cases. Some recent data have suggested that diabetes may occur in patients with Kallmann's syndrome, but further investigation is needed to establish whether gestational diabetes is associated with Kallmann's syndrome or is purely coincidental. PMID- 11727354 TI - In vitro fertilization following natural cycles in poor responders. AB - This prospective study was designed to examine the feasibility of natural cycle in vitro fertilization (IVF) in poor responders, and the clinical factors that may predict successful outcome. Twenty-two poor responders underwent IVF treatment with 44 unstimulated cycles. The results of the natural cycles were compared with those of the 55 low-response stimulated cycles of these patients during the 12 months prior to the study. Eighteen (82%) patients had at least one oocyte retrieved, while nine (41%) had at least one cycle with embryo transfer. Two (9%) patients each gave birth to a healthy term baby. These results are comparable with those of the stimulated cycles. Serum early follicular follicle stimulating hormone (FSH) level was found to be the only reliable predictor of oocyte recovery and overall outcome in each specific natural cycle. However, because of great variability in basal FSH levels among different cycles of the same patient, this is not a reliable predictor of outcome in future cycles. We conclude that poor responders are a unique group of patients who may benefit from natural-cycle IVF treatment. PMID- 11727355 TI - Relationship between estradiol 16 alpha-hydroxylation and human papillomavirus infection in cervical cell transformation. AB - The ovarian steroid hormone estradiol and its metabolite estrone were examined in 45 normal women and 127 premenopausal women with precancerous cervical lesions. Interviews, colposcopy and cervical scrapings were performed. The mean +/- SD values for estradiol and estrone were 0.07 +/- 0.08 ng/ml and 0.06 +/- 0.02 ng/ml, respectively in normal subjects. Corresponding data in patients with cervical intraepithelial neoplasia alone or in association with human papillomavirus (HPV) infection were 0.074 +/- 0.03 ng/ml and 0.076 +/- 0.03 ng/ml or 0.080 +/- 0.03 ng/ml and 0.148 +/- 0.02 ng/ml, respectively, which revealed a significantly greater extent of estrogenic action in the latter population (p < 0.05). We considered that the presence of HPV infection probably increased 16 alpha-hydroxylation of estradiol, providing a possible link between the viral and hormonal elements, possibly having a bearing on the etiology of the disease. PMID- 11727356 TI - Hemostatic effects of high-dose megestrol acetate therapy in patients with advanced gynecological cancer. AB - Effects of high-dose megestrol acetate on blood coagulation and fibrinolysis were investigated in patients with gynecological (n = 13) and breast (n = 10) cancer. Patients received either 160 mg or 320 mg/day megestrol acetate orally. Blood sampling was performed prior to and after months 1, 3 and 6 of treatment. Pretreatment values of global clotting times, fibrinogen, factor VII, thrombin antithrombin III complex, anticoagulation, fibrinolysis and antifibrinolysis were found to be within the reference range. Elevated plasma levels were demonstrated for prothrombin fragments 1 and 2, fibrin degradation products and the plasmin antiplasmin complex. We demonstrated a significant 20-30% reduction of factor VII until the 3rd month of therapy. No further effects were seen within the remaining 3 months of treatment. For the other analyzed parameters of hemostasis, no significant influence of high-dose progestin treatment was found. Furthermore, we observed no clinically relevant differences between the two dosages. Our results do not provide any evidence that there is a thrombogenic effect of high-dosage megestrol acetate with 160 mg or 320 mg per day amongst patients with advanced gynecological malignancies. The observed incidence of thrombosis might be the consequence of other risk factors such as tumor-induced hypercoagulability, simultaneous chemotherapy or other individual thrombosis risk factors. PMID- 11727358 TI - Neuroendocrine effects of raloxifene hydrochloride in postmenopausal women. AB - Raloxifene is a selective estrogen modulator able to exert an estrogen-like action on some target tissues and a specific antiestrogenic action on the uterus and breast. In ovariectomized rats, it has been shown to stimulate the beta endorphin and allopregnanolone concentrations of the anterior and neurointermediate pituitary lobes, the hypothalamus and the hippocampus. The present study aimed to evaluate, in 12 healthy postmenopausal women, the effect of 60 mg/day raloxifene hydrochloride administration for 6 months on plasma beta endorphin and allopregnanolone levels, and on the dynamic changes of both beta endorphin and allopregnanolone secretion after the administration of: (1) clonidine, an alpha 2-presynaptic adrenergic agonist; (2) naloxone, an opioid receptor antagonist; and (3) fluoxetine, a serotonin selective reuptake inhibitor. The administration of raloxifene significantly increased both circulating beta-endorphin and allopregnanolone concentrations, at both the third and sixth months of treatment (p < 0.01). Clonidine, fluoxetine and naloxone administration before therapy was not able to stimulate the release of beta endorphin, but the response was completely restored after raloxifene administration. Before therapy, clonidine and naloxone tests were accompanied by a significant rise in allopregnanolone secretion; the same changes were observed after raloxifene administration, but with significantly higher allopregnanolone concentrations at each time considered. While the fluoxetine test before therapy failed to increase the release of allopregnanolone, the same test after 6 months of raloxifene administration was characterized by a significant release of allopregnanolone at 60 and 90 minutes. The present data indicate that raloxifene has an estrogen-like effect on neuroendocrine pathways in postmenopausal women. PMID- 11727357 TI - A 1-year comparison of the efficacy and clinical tolerance in postmenopausal women of two hormone replacement therapies containing estradiol in combination with either norgestrel or trimegestone. AB - This double-blind, randomized, multi-center study compared the efficacy and clinical tolerance of a combined formulation containing 2 mg estradiol (E2) and 0.5 mg trimegestone (TMG) with a standard hormone replacement therapy containing estradiol valerate (E2V) and norgestrel (NG) in the treatment of climacteric symptoms. The study was conducted over 13 cycles, each of 28 days, and involved 634 subjects, of whom 481 completed the study. The primary efficacy variable was the percentage of subjects who showed at least a 50% reduction from baseline in the mean daily number of hot flushes in cycle 3. This was observed in 98.5% of the subjects in the E2 + TMG group and 93.3% of the subjects in the E2V + NG group (95% confidence interval of the difference, -8.6, -1.9). Significant differences in favor of the E2 + TMG combination were observed in the reduction in the mean daily number and severity of hot flushes, and in the percentage of subjects who had hot flushes at baseline but no hot flushes during treatment. There were no significant differences between the treatments in the Kupperman index and in urogenital signs or symptoms. Treatment with the E2 + TMG combination was well tolerated and the incidences of adverse events were similar in the two treatment groups. Breast pain was the main adverse event, possibly related to treatment that resulted in discontinuation. The mean number of bleeding days per cycle was significantly lower with the E2 + TMG combination than with the E2V + NG combination. The incidences of endometrial hyperplasia were low and comparable in both treatment groups. It was concluded that the E2 + TMG combination was either equivalent or superior to the E2V + NG combination in the treatment of hot flushes and other climacteric symptoms, and that its bleeding profile was favorable. PMID- 11727359 TI - Effects of long-term hormone replacement therapy on arterial wall thickness, lipids and lipoproteins, fibrinogen and antithrombin III. AB - We aimed to study the effects of hormone replacement therapy (HRT) in menopausal women on lipoproteins, blood coagulation, arterial blood pressure and markers of atherosclerosis. A total of 186 postmenopausal women were studied; 98 used HRT (0.625 mg 17 beta-estradiol plus 10 mg progestin) and were followed for 48 months, and were matched with 98 women without therapy. Low-density lipoproteins, high-density lipoproteins, triglycerides, antithrombin III, systolic and diastolic arterial pressure and intima-media thickness were measured at 6, 12, 24, 38 and 48 months. In users of HRT, we found a significant reduction of the lipoproteins, triglycerides, systolic and diastolic pressures and intima-media thickness; however, we found an increase in high-density lipoproteins and anthitrombin III in comparison with non-users. This study has shown a protective effect of HRT on such risk factors and on certain markers of atherosclerosis. PMID- 11727360 TI - Metformin therapy in the management of polycystic ovary syndrome: endocrine, metabolic and reproductive effects. AB - Polycystic ovary syndrome (PCOS) is a convergence of multisystem endocrine derangements. Impairment in insulin metabolism is a prominent feature of the syndrome and appears to play a key pathogenetic role precipitating the cascade of other disorders associated with PCOS. Recent studies report that insulin sensitizing agents, such as metformin, reduce hyperinsulinemia, reverse the endocrinopathy of PCOS and normalize endocrine, metabolic and reproductive functions, leading to the resumption of menstrual cyclicity and ovulation. In this review, we report the most recent evidence regarding metformin usage in women with PCOS. PMID- 11727361 TI - Temporal coupling of luteinizing hormone and follicle stimulating hormone secretion in polycystic ovary syndrome. AB - The aim of this study was to assess the luteinizing hormone (LH) and follicle stimulating hormone (FSH) pulsatile secretion and their temporal relation (concordance) in subjects with polycystic ovary syndrome (PCOS). Fifteen subjects were included in the study (age 17-30 years, body mass index (BMI) 19.38-33.46 kg/m2). For the LH and FSH determinations, blood sampling started at 23.00 and lasted for 6 h with an intersample interval of 10 min. Pulse analysis was carried out using the PulsDetekt program. LH/FSH pulse concordance was calculated using the specific concordance index. Gonadotropin co-pulsatility was found in six subjects who were significantly younger than the others (median 18.5 vs. 22.5 years, p = 0.036). BMI, hirsutism grade, insulin sensitivity, estradiol, progesterone, testosterone, prolactin, cortisol and results obtained from the pulsatility analysis did not significantly differ between the groups. A serum cortisol concentration was correlated with the increased LH/FSH lag time (p = 0.851, p = 0.036) all subjects were included. In conclusion, two distinct LH/FSH secretory patterns were found in PCOS patients, manifested by the presence or absence of the concordance of gonadotropin secretion. In the group where LH/FSH co-pulsatility was present, correlation was found between the serum cortisol and the LH/FSH lag. We also confirmed the finding of previous studies that LH and FSH secretion are regulated by two different mechanisms. PMID- 11727362 TI - Thyroid autoimmunity and infertility. AB - We aimed to study the prevalence of thyroid autoimmunity in infertile women; to assess whether thyroid autoantibodies were associated with non-organ-specific autoantibodies; and to investigate the influence of this dysfunction on the couples' chances of pregnancy. We assayed serum levels of thyroid stimulating hormone (TSH), free thyroxine, and microsomal and thyroglobulin autoantibodies in 149 infertile women. In patients with serum TSH levels in the hypothyroid or hyperthyroid range and/or with thyroid autoantibodies, we performed thyroid ultrasound examinations and assayed some non-organ-specific autoantibodies. We compared the duration of infertility in infertile patients with normal thyroid (control group), with thyroid abnormalities, and with thyroid autoantibodies in euthyroidism. Thirty infertile patients (20.1%) had thyroid abnormalities. The prevalence of thyroid autoantibodies was 17.4%. In infertile patients with thyroid autoantibodies, we found a poor association with non-organ-specific autoantibodies. Only the women with thyroid abnormalities and ovulatory dysfunction had a mean duration of infertility significantly longer than that of the control group. When the data were analyzed for euthyroid women with thyroid autoantibodies, we found no significant variation in the duration of infertility. Although we found a high prevalence of thyroid autoantibodies in infertile patients, the presence of these autoantibodies per se did not reduce the chance of pregnancy. PMID- 11727363 TI - Hypertension in menopausal women--a special case, for special treatment? AB - After the menopause the consequences of hypertension in women change. Their risks of myocardial infarction and stroke rise steeply, a rise that has been blamed in part on the loss of estrogen and the onset of menopausal metabolic syndrome, with endothelial dysfunction, hyperlipidemia, insulin resistance and derangement in coagulation. Hypertensive menopausal women have not had optimum treatment. They have poorer prognoses than men of the same age. Their antihypertensive management therefore merits special attention. Hormone replacement, aspirin prophylaxis and lipid-lowering drugs have their place. The antihypertensive drug chosen should not worsen the metabolic syndrome: angiotensin-II converting enzyme (ACE) inhibitors are therefore among the first-line drugs. Few drugs have been specifically aimed at menopausal hypertension and these are reviewed here. PMID- 11727364 TI - Technology after the fall. PMID- 11727365 TI - Communications that won't quit. PMID- 11727366 TI - Security you can live with. PMID- 11727367 TI - An update on the treatment of gait problems in cerebral palsy. AB - This article summarizes our experience with cerebral palsy over the past 20 years. The primary and secondary deformities that occur with cerebral palsy are described. Following this, there is a brief overview of the nature and role of gait analysis in the treatment of gait problems in cerebral palsy. The concept of lever-arm dysfunction is introduced. Our current treatment algorithm is then presented along with a brief discussion of our current treatment program, which is illustrated by a case example. Finally, a brief study of a group of patients with spastic diplegia or quadriplegia is presented to illustrate our current method of evaluating treatment outcomes and the need for team management in the treatment of this complex condition. PMID- 11727368 TI - Prediction of postoperative gait velocity in cerebral palsy. AB - Preoperative and postoperative gait analyses were reviewed for 47 patients with cerebral palsy in an attempt to ascertain predictors of surgical outcome as measured by gait velocity. Higher postoperative velocity correlated with higher preoperative velocity and younger age. Observed velocities in older children were smaller than predicted values. Older children showed either an average decline in velocity from normal for age or no change. Younger children showed an average increase in velocity toward normal for their age. Diagnosis, type of surgery, number of procedures performed and level of ambulation preoperatively were not predictive of postoperative velocity. These results suggests that gait velocity cannot be reliably increased in all children with cerebral palsy undergoing surgery, especially those older than age 12. PMID- 11727369 TI - Varus derotation osteotomy for the treatment of hip subluxation and dislocation in cerebral palsy: statistical analysis in 73 hips. AB - The purpose of this study is to review the results of varus osteotomy in patients with cerebral palsy and to determine factors that influence the rates of residual hip displacement. A retrospective chart review of 65 patients who underwent 79 varus osteotomies was performed. Preoperative, postoperative and follow-up radiographs were analyzed for routine radiographic measurements, pelvic obliquity, osteonecrosis (avascular necrosis), joint incongruity or degenerative joint disease. The average follow-up was 5.2 years (range, 1.1-18.4 years). At follow-up, 3 hips were dislocated, 19 were subluxated and 57 were stable (72%). Age at surgery and the degree of preoperative hip displacement had significant effects on outcome. The average age at surgery for initially subluxated hips, which were located at follow-up, was 7.2 years. This was significantly younger (P = 0.008) than initially subluxated hips, which were displaced (10 years of age). Subluxated hips at surgery were also more likely to be located at follow-up than dislocated hips. PMID- 11727370 TI - Conservative management of dynamic equinus in diplegic children treated by gait improvement surgery. AB - Multilevel surgery for gait improvement was performed on twelve ambulatory children with diplegic type of cerebral palsy and dynamic equinus deformity. Dynamic equinus deformities were defined as those who had an equinus at initial contact during preoperative gait analysis, and where the equinus deformity was correctable passively during physical examination. Ankle function was evaluated by clinical examination and gait analysis before surgery, and at least 3 years after surgery. The ankle showed an increase in dorsal flexion at initial contact, at single stance and in swing. There was an increase in dorsal flexion at the beginning of push-off, without a decrease in the range of motion of the ankle during push-off. Ankle moments demonstrated significant improvement in the maximum flexor moment in the second half of single stance. There was a change from abnormal generation of the energy in mid-stance to the normal pattern of energy absorption. Positive work during push-off was significantly increased. Conservative management of dynamic equinus deformities combined with multilevel surgery to correct other deformities of the locomotion system resulted in significant improvements in ankle function during gait. PMID- 11727371 TI - Reliability of the Severin classification in the assessment of developmental dysplasia of the hip. AB - This study was undertaken to investigate the reliability of Severin's classification at various ages, and to determine whether the reliability is improved by careful measurement rather than subjective assessment. The radiographs taken at ages 6, 10 and 16 years of 20 randomly selected patients treated for developmental dysplasia of the hip were graded by six observers on two separate occasions using parameters measured according to Severin's criteria. In addition, four of these observers regraded the same radiographs using subjective assessment without measurements being made on two other separate occasions. Agreements between and within observers were evaluated using the weighted Cohen's kappa statistic for each age group. Intraobserver reliability was good, there being a close association between the measured and the subjectively observed ratings. This accords with the subjective nature of this classification. The interobserver reliability was found to be poor although it improved when direct measurements were made. Overall agreement between observers improves as patient age increases. It is concluded that comparisons between different observers using the Severin classification system are not reliable. However, a single investigator comparing treatment modalities in the same study allowing for individual bias in assessing deformity and subluxation would produce reliable results. PMID- 11727372 TI - Sequential magnetic resonance imaging in slipped capital femoral epiphysis: assessment of preslip in the contralateral hip. AB - Thirty-three images using magnetic resonance imaging (MRI) in 10 patients with slipped capital femoral epiphysis were evaluated. The MRI of affected hips delineated typical widening of the physis. Sequential MRI demonstrated the process of premature closure of the physis developing from the posterior portion anteriorly. In 4 of the 10 asymptomatic contralateral hips, physeal widening was clearly observed in the center or posteromedial region of the physis on T1 weighted images. We performed prophylactic pinning for the four cases showing physeal widening on MRI, and none of the other cases developed a slip. MRI accurately documented detailed features of slips, as well as physeal changes, and may provide indicative information on prophylactic pinning for contralateral hips. PMID- 11727373 TI - Coxa vara: another option for fixation. AB - Coxa vara is a progressive childhood deformity that is best remedied by a valgus intertrochanteric osteotomy. By restoring the femoral neck and its physis to a more anatomic position, hip mechanics are normalized, with consequent beneficial effects upon hip motion, limb lengths and acetabular development. In the pediatric age group, there is no implant specifically designed for this purpose. Therefore, in addition to the challenge of obtaining adequate correction, inadequate fixation may contribute to recurrent varus deformity, a problem that, according to the literature, has a 30% to 70% prevalence. We are presenting a new technique using a modified veterinary plate for osteosynthesis in children in the 5-year-old to 11-year-old age group. The rationale, implant design, surgical technique and results in a group of nine patients (12 hips) are described. PMID- 11727374 TI - Knee arthroscopy in children and adolescents. AB - We report a series of 77 skeletally immature patients (41 children aged < or = 13 years and 36 adolescents aged 14-16 years) who underwent knee arthroscopy. The purpose of the study was to assess the accuracy of clinical diagnosis and the value of arthroscopy. The correlation between clinical and arthroscopic diagnoses was analyzed in 54 patients. Arthroscopy confirmed the clinical diagnosis in 46.29% (25) of the patients. The accuracy of clinical diagnosis in patients aged 13 years and younger was 31.25%, whereas the accuracy in patients aged from 14 years to 16 years was 68.18%. Paediatric knee arthroscopy is a safe procedure that significantly improves the accuracy of clinical diagnosis, allows the definitive treatment of a variety of conditions and often prevents an unnecessary arthrotomy. PMID- 11727375 TI - Does functional treatment of idiopathic clubfoot reduce the indication for surgery? Call for a widely accepted rating system. AB - We present our experience of treatment by physiotherapy, continuous passive motion and strapping in a series of 100 clubfeet classified on a scale of severity according to Dimeglio. Twenty-five percent were good after conservative treatment, and 75% required an operation. There were no recurrences or additional procedures. Before the introduction of the functional treatment in our department, 100% required some sort of surgical intervention and 51% required an additional procedure. In comparison with the results published by Dimeglio et al., the greatest discordance is observed in grade 2 clubfeet. PMID- 11727376 TI - Traumatic atlanto-occipital dislocation with survival in children. AB - Traumatic atlanto-occipital dislocation is a rarely seen clinical entity, because it is too often fatal. The case of an 8-year-old boy is presented with an anterior atlanto-occipital dislocation. He suffered from multiple injuries: severe head injury and coma, fractures of both femurs and one tibia. He was considered an incomplete spinal cord injury on admission. The diagnosis was suspected 4 days after admission, and a minimal cervical traction was applied for 4 weeks. At 5 years follow-up, clinical and radiologic status was normal. This case is compared with the 28 survivor children reported in the literature: 79% had incomplete neurologic lesions, and only 12 were normal at follow-up. Every report pointed out difficulties of initial clinical and radiologic diagnosis. Treatment is varied and controversial but, in children's cases, mild traction may be indicated in anterior and posterior displacement as initial management, and surgical stabilization performed early in longitudinal dislocation and deferred in displacement that remains unstable. PMID- 11727377 TI - Mobile thoracolumbar gibbus in Morquio type A: the cause of paraparesis and its management. AB - Four children with Morquio A syndrome had neurologic deterioration related to a progressive thoracolumbar gibbus. Findings on the magnetic resonance imaging scan did not suggest a significant canal compromise, but X-rays taken in the erect position showed a much larger kyphosis, and thus more canal compromise. All patients recovered neurologically after anterior decompression and correction of kyphosis. We believe that the cause of cord dysfunction is mobility at the kyphus and compression by the bulging discs and the internal gibbus. PMID- 11727378 TI - The 'upside-down' radial head: a diagnostic challenge. AB - The author describes a rare, traumatic dislocation of the radial head in an 8 year old girl. Only four similar cases were found in the literature. The danger of misdiagnosis and the life-long disability can be avoided by careful examination of the post-reduction radiograph. PMID- 11727379 TI - Histomorphometric study of growth plate subjected to different mechanical conditions (compression, tension and neutralization): an experimental study in lambs. Mechanical growth plate behavior. AB - We studied the morphologic effect of low mechanical stresses (compression, tension and neutralization) on the growth cartilage with an external fixator in 18 young lambs. On radiography, we only found more length in the femora subjected to tension (P < 0.05). Bony bridges were not present, nor were there signs of altered vascularization or Ranvier's perichondral ring. Histomorphometrically, in the group subjected to tension, the germinative layer in the femur was higher (P < 0.001) in the bones that had undergone surgery. The proliferative layer was lower in the operated tibia (P < 0.001), and the hypertrophic layer was higher in operated tibias and femora (P < 0.001). In the group subjected to compression, the germinative layer in the femora was higher in the operated bones (P < 0.05); the proliferative layer of the tibia was lower (P < 0.001) and the hypertrophic layer was higher (P < 0.001) in both operated bones. In the neutralization group, the proliferative layer of both operated bones presented lower values (P < 0.001) and the hypertrophic layer was higher (P < 0.001) than that in the control groups. PMID- 11727380 TI - Response of the physis to leg lengthening. AB - The physeal response to limb lengthening by callotasis was investigated in an animal model by measuring 1) the growth rate using an oxytetracycline labelling technique, 2) histomorphological changes, and 3) proliferative cellular activity using bromodeoxyuridine. An osteotomy without lengthening stimulated growth at the physis. At 20% lengthening, regardless of the rate of distraction, no significant changes were detected. However, the growth rate was drastically reduced after 30% or more of lengthening and in 50% of cases there was evidence of premature closure of the distal growth plate. CLINICAL RELEVANCE: Caution should be exercised in carrying out high percentages of lengthening of children's bones. PMID- 11727381 TI - Madelung deformity: clinical features, therapy and results. AB - In a retrospective study, 31 patients with Madelung deformity were reviewed. They were treated at one institution during a period of 15 years. On first presentation, the mean age was 22.5 years with a range from 10 years to 64 years. Twenty-four patients (77%) were female. The main complaints were pain, limited range of motion, and objectionable appearance. A family history of Madelung deformity was present in four patients (13%). The diagnosis of Leri-Weill syndrome could not be confirmed in any case. There was no correlation between the clinical appearance and the extent of radiologic abnormality. Five patients (16%) were operated on because of permanent pain. On postoperative examination, only one patient revealed no restricted range of mobility and no pain, whereas the other four patients improved in terms of pain but showed only limited improvement of function. The vast majority of patients, however, required no surgical therapy. PMID- 11727382 TI - Bilateral wrist dislocation in trisomy 21: a case report. AB - Ligamentous laxity is common in Trisomy 21 (Down's syndrome). The literature has numerous reports of atlanto-axial instability, patellar instability, carpal instability and other manifestations of joint instability. To date, no report has been published in the English literature describing non-traumatic unilateral or bilateral wrist dislocation either in trisomy 21 or in any other condition. PMID- 11727383 TI - Grip strength in children. AB - The grip strength in a group of 487 healthy children aged between 5 years and 15 years was measured. There was a clear correlation between age and grip strength. Up to the age of 12 years, there was a striking parallellism between boys and girls. From the age of 13 years, boys developed a 25% stronger grip force than girls. The difference between the dominant and nondominant hand grip was not significant, but was constantly stronger in the right hand for right-hand dominant children. There was a high variation in grip forces (from 6.9 kg to 38.4 kg). PMID- 11727384 TI - Techniques for small-bone lengthening in congenital anomalies of the hand and foot. AB - The purpose of this study is to analyse three different lengthening techniques used in 31 small bones for congenital malformations of the hand and foot: 15 metacarpals, 12 metatarsals, 1 foot stump and 3 spaces between a previously transplanted phalanx end of the carpus or the metacarpal. Progressive lengthening with an external fixator device was performed in 23 cases: the callus distraction (callotasis) technique was used in 15 cases, whereas in the other 8 cases the speed of lengthening was faster and the defect bridged with a bone graft as a second stage. In another eight cases, a one-stage lengthening was performed. In the callotasis group, the total length gained ranged from 9 mm to 30 mm and the percentage of lengthening obtained (compared with the initial bone length) averaged 53.4%; in the fast lengthening group, the length gained ranged from 8 mm to 15 mm, and the average percentage of lengthening was 53.1%; and in the one stage group, the length gained ranged from 7 mm to 15 mm, and the average percentage of lengthening was 43%. The overall complication rate was 22.5%. PMID- 11727385 TI - Primary chronic sclerosing (Garre's) osteomyelitis in children. AB - Three children with unifocal nonpyogenic inflammatory bony lesions with a prolonged, fluctuating course are reported. The lesions were located at the metaphyseal region of long bones. Three was progressive sclerosis and hyperostosis in the tibia or femur, such as the changes described in Garre's osteomyelitis. No pus was released by exploration of the lesions. Tissue and blood cultures were negative. The histology was typical of chronic osteomyelitis: the symptoms returned intermittently over several years, together with the development of sclerosis but without disturbance of bone growth. It is not clear whether Garre's chronic sclerosing osteomyelitis is a different entity from chronic recurrent multifocal osteomyelitis. PMID- 11727386 TI - Establishing the validity of test score inferences: performance of 4th-year U.S. medical students on the ECFMG Clinical Skills Assessment. AB - BACKGROUND: Although the relations among prematriculation, matriculation, and residency assessments have been studied, measures pertaining to clinical skills have rarely been incorporated in any analyses. PURPOSE: The purpose of this article was to investigate the relations between scores obtained on the Educational Commission for Foreign Medical Graduates Clinical Skills Assessment (CSA) prototype and aptitude-ability measures designed to select medical students, to assess medical students while in medical school, and to evaluate physicians in postgraduate training programs. The sample included 122 4th-year medical students. METHODS: Relations among scores were summarized with correlation coefficients. Analysis of variance was used to compare CSA scores by departmental grades. RESULTS: Significant correlations (p < .01) were found between scores from assessments commonly used in medical school (i.e., United States Medical Licensing Exam [USMLE] Steps 1 and 2) and CSA component scores. There were weak associations between CSA measures and both Medical College Admissions Test scores and residency program evaluations. CONCLUSIONS: The relations between CSA scores and various other medically oriented ability measures provide additional evidence that inferences based on CSA scores are appropriate and valid. PMID- 11727387 TI - National survey of internal medicine residency programs of their 1st-year experience with the electronic residency application service and national resident match program changes. AB - BACKGROUND: The residency recruitment and selection process is a critical one for residency programs and medical students. In 1999, internal medicine programs conducted the residency match on the Web for the first time using the Electronic Residency Application Service (ERAS). PURPOSE: The authors wished to study the impact of this change on house staff recruitment and quality of match. METHOD: A Web-based survey with e-mail, paper, and fax reminders was sent to all 407 internal medicine residency programs after the 1999 match. RESULTS: Eighty-six percent of reporting programs found the screening of applicants easier. The overall number of applicants varied greatly (48% of programs reported more applicants; 32% reported fewer). The quality of final match was rated the same as previous years by 47%, better by 38%, and worse by 15%. CONCLUSIONS: The transition to ERAS was successful in internal medicine. However, there are several areas that were identified that will improve the ERAS process as it evolves. PMID- 11727388 TI - Standardized patients' perceptions about their own health care. AB - BACKGROUND: There have been detailed descriptions on standardized patients (SP) programs' effects on students, curricula, and faculty, yet little attention has been paid to the consequences of participating on the SP's. PURPOSE: This study explored the perceptions of SPs toward their own health care in the context of having served as SPs. METHOD: All 180 SPs participating in Department of Medicine programs at 5 medical schools were surveyed. They completed the survey during SP activities, or it was mailed to them. SPs indicated their level of agreement or disagreement with 11 attitude statements related to their own health care after serving as an SP using a Likert scale, with 1 reflecting the most positive attitude and 5 the least positive. RESULTS: Responses to the attitudinal questions were obtained from 164 SPs (91%). SPs perceived that because of their participation as SPs they had a better understanding about medical history taking and physical examinations (1.9 +/- 0.9), communicated more effectively with their health care provider (1.8 +/- 0.9), and were more comfortable with both health care visits and physical examinations (2.2 +/- 0.9). There were no significant differences in results based on gender, age, race, or school. CONCLUSIONS: As a consequence of their participation, the SPs indicated a change in attitudes about their personal health care. They perceived improved understanding and ability to communicate and comfort with their own health care. Participation in SP programs seems to influence SPs by improving perceptions about their own health care interactions. PMID- 11727389 TI - Personalized remedial intensive training of one medical student in communication and interview skills. AB - BACKGROUND: Medical students found to be deficient in communication and interview skills pose a difficult remediation challenge. There is no standard way to address such deficiencies. The authors describe the development and implementation of an intensive remedial curriculum. DESCRIPTION: A 2nd-year student found deficient in communication skills was held back for a year and paired with a clinical preceptor for intensive skills training, including a weekly precepted clinic, structured readings, standardized patient (SP) exercises, communications workshops, and end-of-year standardized clinical evaluations. EVALUATION: The student's self-assessment and the preceptor's assessments of communication skill gradually improved over the year. The student improved through a progression of SP exercises focusing on specific communication skills. The student passed a final evaluation exercise with an excellent rating from the SP and the preceptor. CONCLUSIONS: This multistrategy approach to improve communication skills can be applied to other students and in other institutions. Based on the Bayer Institute communication workshops and integrated coaching techniques, the material forms a framework to help deficient students to become proficient in communication and interviewing skills. PMID- 11727390 TI - First aid and basic life support skills training early in the medical curriculum: curriculum issues, outcomes, and confidence of students. AB - BACKGROUND: The importance of training physicians and other health care professionals in first aid and basic life support (BLS) is now widely recognized. The Faculty of Medicine and Health Sciences of the United Arab Emirates University have been offering formal training in these skills to 1st-year medical students since 1989. The aim of this study was to determine the efficacy of the program and the extent to which it provides students with confidence to perform skills on their own after training. DESCRIPTION: 165 of 180 male and female students comprising 3 cohorts (91.7% response rate) completed a self-administered anonymous questionnaire. The questionnaire contained open- ended and Likert-type questions regarding course organization, content, strengths and weaknesses, teaching--learning approaches, and skills development and confidence. The authors analyzed assessment outcomes and associations between self-perceived levels of confidence and ability to practice. EVALUATION: Students were uniformly enthusiastic and highly motivated by the program. Self-assessed confidence in ability to perform skills on their own after completing the program was moderately correlated with perceived frequency of opportunity to practice many skills. There was nevertheless a consistent desire for more practice time. CONCLUSIONS: The results suggest that the program provides students with sound basic knowledge and adequate practical skills in first aid and BLS and that adequacy of time and physical and human resources are important prerequisites to facilitate practice and engender confidence in skills. PMID- 11727391 TI - Interdisciplinary education in a community-based geriatric evaluation clinic. AB - BACKGROUND: Demographic predictions are challenging schools of medicine to emphasize geriatrics. This article describes a geriatric evaluation (GET) clinic and explores the opportunity for residents to attain core geriatric competencies. DESCRIPTION: The GET clinic is located in a small Midwestern city associated with a community-based medical school. It is staffed by an interdisciplinary team consisting of a geriatrician, a gerontological nurse specialist, and a social worker. Residents, medical students, and nursing students are frequent participants. EVALUATION: Descriptive data indicate that the clinic experience addresses the core competencies set forth by the American Geriatric Society. The clinical outcomes indicate that the clinic is effective in maintaining the functional status of patients and has a positive effect on family caregivers. Residents and students rate their experience as excellent. CONCLUSIONS: The GET clinic provides a unique interdisciplinary educational opportunity. Further investigation is needed to determine if residents who participate do attain core competencies. PMID- 11727392 TI - Learning styles and perceptions of the value of various learning modalities before and after a 2nd-year course in microbiology and infectious diseases. AB - BACKGROUND: The authors' 4-week course in microbiology and infectious diseases consists of lectures, small-group sessions, interactive computer-assisted learning (CAL), and textbook readings. PURPOSE: To determine how individual learning style influenced learners' value assessment of these teaching modalities. METHODS: A Kolb Learning Style Inventory and questionnaire to assess enthusiasm for each teaching modality were administered before the course. At course end, a 2nd questionnaire assessed the perceived usefulness of each teaching modality. RESULTS: Learners with a relative preference for experiential learning rather than abstraction initially favored small groups (R2 = .06, p = .004) and CAL (R2 = .06, p = .005). Similarly, learners with a preference for reflective observation rather than active experimentation favored lectures (R2 = .05, p = .01). However, at course end, Kolb learning style did not predict the value assessment of any modality. CONCLUSIONS: Kolb learning style influenced the initial attractiveness but not the retrospective assessment of learning modalities; hence, quality and content superseded learning style as determinants of value after course completion. PMID- 11727393 TI - Offices of research in medical education: accomplishments and added value contributions. AB - PURPOSE: Offices of research in medical education have been in existence since the 1st one was begun by Hale Hamm at Case Western Reserve in 1958. There are now 61 medical schools in North America that have a formal office of medical education (OME) and are part of the Society of Directors of Research in Medical Education (SDRME). The purpose of this study was to report how SDRME and OMEs have contributed to the research in medical education (RIME) efforts. SUMMARY: This study reports specific initiatives that have been begun by the SDRME culled from historical documents and business meeting minutes and profiles results of the 2000 biannual membership survey that describe what the individual units are doing in support of their institutions. As an organization, SDRME sponsors literature reviews, 5 of which have been published in peer-reviewed journals. It has also collaborated with the American Medical Association, the National Institutes of Health Bureau of the Health Professions (NIH BHP), and the Association of American Medical Colleges in offering conferences and workshops. SDRME worked with the Macy Foundation on a grant initiative which by November 1993 had established 6 consortia to develop regional centers on clinical competence and with the NIH BHP in the development of a grants program that established centers for RIME. Results from the 2000 membership survey indicate that individual units are heavily tied into evaluation efforts at both the institutional and national levels. Research efforts being engaged by these units have led to a reasonably high, but variable, level of publication activity. The current wave of new curricula being implemented in medical education centers are being heavily supported by these units, with all units being involved in curriculum planning and administration. CONCLUSIONS: SDRME has been effective in promoting RIME through sponsoring literature reviews and collaborating with various national organizations. OME units have been effective in serving the needs of their institutions as well as contributing to national research efforts. PMID- 11727394 TI - The managed care pharmacy perspective. AB - Despite the important reproductive and noncontraceptive benefits of hormonal therapy, both oral contraceptives (OCs) and hormone replacement therapy (HRT) are underutilized, with only a small portion of eligible women receiving therapy. Increased use of hormonal therapy will result in greater pharmacy costs, a concern in the present era of cost containment that is reflected in the wide variability in coverage of hormonal therapy provided by managed care organizations. However, pharmacoeconomic research demonstrates that relatively small expenditures in pharmacy costs for hormonal therapy result in significantly lower healthcare costs per patient, through the prevention of unintended pregnancy with OCs and the noncontraceptive health benefits of both OCs and HRT. PMID- 11727395 TI - Pharmacologic characteristics of progestins used for contraception and hormone replacement therapy, including new transdermal technologies. AB - Structural differences among progestins account for their unique properties. Pharmacologic principles should be used when considering the appropriate progestational agent to achieve both clinical and economic outcomes. Use of the same progestin in oral contraceptives (OCs) and hormone replacement therapy (HRT) allows for a seamless transition from OCs to HRT, while maintaining the health benefits of the OC. A pharmacologically based approach also has been used in developing the first transdermal contraceptive system, which is expected to be on the market in early 2002. PMID- 11727396 TI - Managing the transition from oral contraceptives to hormone replacement therapy. AB - Oral contraceptives provide multiple benefits for perimenopausal women; their use results in considerable economic savings by managing vasomotor symptoms, preventing pregnancy, and improving future outcomes. Hormone replacement therapy provides many health benefits for menopausal women; a recently introduced constant estrogen/intermittent progestin regimen takes advantage of receptor dynamics and the extended half-life of norgestimate to provide a continuous progestational effect on the endometrium. The lower progestin dose may also decrease progestin-related side effects and contribute to increased adherence and treatment continuation. PMID- 11727397 TI - Patient communication and counseling on contraception and hormone replacement therapy. AB - Many women are unaware of the wide range of benefits resulting from hormonal therapy. Clinicians need to improve their ability to counsel patients about the benefits and risks of oral contraceptives (OCs) and hormone replacement therapy to improve patient compliance and willingness to continue therapy. For reproductive-age women, choosing the lowest effective estrogen/progestin dose will reduce the side effects and bleeding irregularities that are major reasons for patient discontinuation. Analysis has shown that compared with OCs the new transdermal contraceptive system, which is expected to be on the market in 2002, has better outcomes and lower costs at all ages. PMID- 11727398 TI - Treatment for the procoagulant state in type 2 diabetes. AB - A procoagulant state has been found to exist in diabetes mellitus. There may be activation of the intrinsic coagulation system, decreased fibrinolytic activity, or alterations in platelet function. Intensive glycemic control with insulin is effective in reducing the impact of this procoagulant state by favorably affecting all three components of the system. Decreased fibrinolytic activity, as influenced by plasma PAI-1 levels, may be favorably affected by weight loss, exercise, a low-GI diet, or by metformin, thiazolidinediones, gemfibrozil, and ACE inhibitor therapy. Insulin has variable effects on plasma PAI-1 activity. Estrogens will lower the elevated PAI-1 levels seen in the menopausal state. Collaborative trial evidence supports the use of low-dose aspirin as a primary or secondary prevention strategy in diabetic persons who are at high cardiovascular risk. A recent study suggests that this category includes virtually every type 2 diabetic individual in the United States. The American Diabetes Association recommends enteric-coated aspirin, 81 to 325 mg/day, as the first choice. In the case of aspirin allergy, clopidrogel is an alternate choice. Thus, recognition of and therapy for a procoagulant state in diabetes mellitus is likely to result in a decrease in the atherothrombotic events that characterize the later stages of this disease. PMID- 11727399 TI - Cardiomyopathy and heart failure in diabetes. AB - Patients with insulin resistance or type 2 diabetes have a particularly high risk for heart failure and a poor prognosis once they develop heart failure. The choice of drugs for the management of heart failure in these patients should be directed at changing the natural history of the disease. The various drugs available for the treatment of heart failure, including ACE inhibitors and beta adrenergic blockers, are known to be beneficial and should be given as first-line agents. Aggressive risk-factor modification and tight blood pressure and glycemic control are crucial. Much work is needed to establish the safety and efficacy of various oral antidiabetic agents, especially the TZDs, for which the theoretic benefits are substantial and overall morbidity and mortality impact remain ill defined. PMID- 11727400 TI - Pathogenesis of type 2 diabetes. Insulin resistance. AB - This article addresses the role of insulin resistance in the pathogenesis of type 2 diabetes. The major causes of acquired insulin resistance including aging, pregnancy, lack of physical activity, and obesity are discussed briefly. The role of free fatty acids (FFAs) as a link between obesity and insulin resistance/type 2 diabetes is discussed in detail. Evidence is provided showing that increased plasma FFA levels produce insulin resistance dosage dependently, acutely, and chronically. Mechanisms by which FFA can cause insulin resistance are outlined. Lastly, normalizing plasma FFA levels is proposed as a new approach to reducing insulin resistance and the risk for type 2 diabetes mellitus, and atherosclerotic vascular disease. PMID- 11727401 TI - Hyperglycemic crises in diabetes mellitus type 2. AB - Hyperglycemic crises in type 2 diabetes are not rare and are becoming increasingly recognized as part of the spectrum of the presentation of previously undiagnosed diabetes mellitus and the decompensation of established diabetes mellitus. Contributing factors and associations are being elucidated but remain far from clear, particularly in DKA states. Medications commonly used in the treatment of many comorbid illnesses in patients with diabetes can themselves predispose to HHS. Endocrinopathies can contribute to insulin resistance and directly increase the glycemic load, leading to hyperglycemia. Medications such as the protease inhibitors may in the future lead to a better understanding of the pathophysiology of the metabolic derangements seen in the development of type 2 diabetes mellitus. PMID- 11727402 TI - Microvascular complications. Retinopathy and nephropathy. AB - Diabetic retinopathy and diabetic nephropathy extract an enormous toll on patients with diabetes and an enormous burden on the health care system. With aggressive control of glycemia and blood pressure, coupled with aggressive use of laser photocoagulation and treatment of microalbuminuria, these problems can largely be eliminated. In the future, specific interventions may emerge that will allow interdiction of the pathophysiologic processes that lead to initiation and progression of these microvascular complications. The challenge for the primary care physician and diabetologist is to attain excellent glycemic control and aggressive control of blood pressure, while assuring that every patient has appropriate dilated fundus examinations at least annually, preferably by an ophthalmologist or retinal specialist, and regular screening for microalbuminuria. With such medical management, appropriate intervention can occur to reduce the risk of blindness and renal failure and to lessen the burden from diabetic retinopathy and nephropathy. PMID- 11727403 TI - Diabetes mellitus and coronary heart disease. AB - Diabetes mellitus, especially type 2 diabetes, is a growing concern in America. Longitudinal trends show that obesity is more prevalent than in the past, and the incidence of type 2 diabetes is also increasing. Type 2 diabetes typically doubles the CHD risk in men and triples the risk in women. Intervening to control lipid levels and blood pressure has been shown to be especially helpful in preventing CHD, but the impact of better glycemic control on CHD risk is less convincing, especially in clinical trials. Revascularization studies in diabetics show that coronary bypass surgery is related to better outcomes than angioplasty procedures. PMID- 11727404 TI - Diet and exercise in type 2 diabetes mellitus. AB - The question is no longer whether diet and exercise can benefit the individual with type 2 diabetes. Rather, the type and duration of exercise the magnitude of the effects on glycemic control, insulin sensitivity, and on risk factors for cardiovascular disease must be considered in determining the feasibility and acceptability of an intervention program. It is now clear that regular physical exercise is important in both the prevention and treatment of type 2 diabetes. The benefits of exercise are many and include increased energy expenditure, which, combined with dietary restriction, leads to decreased body fat, increased insulin sensitivity, improved long-term glycemic control, improved lipid profiles, lower blood pressure, and increased cardiovascular fitness. Persons with type 2 diabetes often find it difficult to exercise and are at increased risk for injury or exacerbation of underlying diseases or diabetic complications. Therefore, before starting an exercise program, all patients with type 2 diabetes should have a complete history and physical examination, with particular attention to evaluation of cardiovascular disease, medications that may affect glycemic control during or after exercise, and diabetic complications including retinopathy, nephropathy, and neuropathy. Exercise programs should be designed to start slowly, build up gradually, and emphasize moderately intense exercise performed at least three times a week and preferably five to seven times a week for best results. PMID- 11727405 TI - Oral therapies for diabetic hyperglycemia. AB - Many classes of oral antihyperglycemic agents are available for the treatment of type 2 diabetic patients. These classes improve glucose metabolism by different mechanisms, and their effects are additive. Therapy with lifestyle modification and a single oral antihyperglycemic agent infrequently achieves target glycemic goals, and, if it does, the effect is usually not sustained. A more rational approach would seem to be therapy with combinations of drugs with different mechanisms of action. Initial therapy might be with submaximal concentrations of two drugs. As the diabetic abnormalities progress, maximal concentrations of the drugs and addition of other classes of oral agents or insulin may be needed to maintain the target glycemic goal. In choosing combinations of oral antihyperglycemic agents, their effects on the components treatment of type 2 diabetic patients. These classes improve glucose considered, as must the specific effects of the agents on glucose metabolism. PMID- 11727406 TI - Insulin therapy in type 2 diabetes. AB - Type 2 diabetes is a common disorder often accompanied by numerous metabolic abnormalities leading to a high risk of cardiovascular morbidity and mortality. Results from the UKPDS have confirmed that intensive glucose control delays the onset and retards the progression of microvascular disease and possibly of macrovascular disease in patients with type 2 diabetes. In the early stages of the disease, insulin resistance plays a major role in the development of hyperglycemia and other metabolic abnormalities, and patients with type 2 diabetes often benefit from measures to improve insulin sensitivity such as weight loss, dietary changes, and exercise. Later, the use of oral insulin secretagogues and insulin sensitizers as monotherapy and in combination helps maintain glycemia for varying periods of time. Ultimately, because of the progressive nature of the disease and the progressive decline in pancreatic beta cell function, insulin therapy is almost always obligatory to achieve optimal glycemic goals. Not all patients are candidates for aggressive insulin management; therefore, the goals of therapy should be modified, especially in elderly individuals and those with co-morbid conditions. Candidates for intensive management should be motivated, compliant, and educable, without other major medical conditions and physical limitations that would preclude accurate and reliable HGM and insulin administration. In selected patients, combination therapy with insulin and oral antidiabetic medications can be an effective method for normalizing glycemia without the need for rigorous multiple-injection regimens. The patients for whom combination therapy is most commonly successful are those who do not achieve adequate glycemic control using daytime oral agents but who still show some evidence of responsiveness to the medications. Bedtime intermediate-acting or predinner premixed intermediate- and rapid-acting insulin is administered and progressively increased until the FPG concentration is normalized. If combination therapy is not successful, a split-mixed regimen of intermediate- and rapid-acting insulin equally divided between the prebreakfast and pre-dinner periods is advised for oese patients, and more intensive regimens are advised for thin patients. Insulin therapy is invariably associated with weight gain and hypoglycemia. The use of metformin or glitazones in combination with insulin has been demonstrated to have insulin-sparing properties. Also, metformin use may ameliorate weight gain. The use of continuous subcutaneous insulin infusion pumps can be particularly beneficial in treating patients with type 2 diabetes mellitus who do not respond satisfactorily to more conventional treatment strategies. Intraperitoneal insulin delivery systems hold considerable promise in type 2 diabetes because of their more physiologic delivery of insulin and their ability to inhibit hepatic glucose production selectively, with less peripheral insulinemia than with subcutaneous insulin injections. Newer insulin analogues such as the rapidly acting Lispro insulin and the peakless, long-acting glargine insulin are increasingly being used because of their unique physiologic pharmacokinetics. New developments such as inhaled and buccal insulin preparations will also make it easier for many patients to initiate and maintain a proper insulin regimen. Finally, a new generation of gut peptides such as amylin and GLP-1 will add a new dimension to glycemic control through modification of nutrient delivery and other mechanisms; however, the ultimate goal in the management of type 2 diabetes is the primary prevention of the disease. The Diabetes Prevention Program (DPP) sponsored by the National Institutes of Health has currently randomly assigned more than 3000 persons with impaired glucose tolerance and at high risk of developing diabetes into three treatment arms: metformin arm, an intensive lifestyle-modification arm, and a placebo arm. The study will conclude in 2002 after all participants have been followed for 3 to 6 years. PMID- 11727407 TI - Pathophysiology of hypertension and endothelial dysfunction in patients with diabetes mellitus. AB - There is compelling evidence for endothelial dysfunction in both type 1 and type 2 diabetics. This dysfunction is manifest as blunting of the biologic effect of a potent endothelium-derived vasodilator, nitric oxide, and increased production of vasoconstrictors such as angiotensin II, ET-1, and cyclooxygenase and lipoxygenase products of arachidonic acid metabolism. These agents and other cytokines and growth factors whose production they stimulate cause acute increases in vascular tone, resulting in increases in blood pressure, and vascular and cardiac remodeling that contributes to the microvascular, macrovascular, and renal complications in diabetes. Reactive oxygen species, overproduced in diabetics, serve as signaling molecules that mediate many of the cellular biochemical reactions that result in these deleterious effects. Adverse vascular consequences associated with endothelial dysfunction in diabetes mellitus are Decreased nitric oxide formation, release, and action Increased formation of reactive oxygen species Decreased prostacyclin formation and release Increased formation of vasoconstrictor prostanoid Increased formation and release of ET-1 Increased lipid oxidation Increased cytokine and growth factor production Increased adhesion molecule expression Hypertension Changes in heart and vessel wall structure Acceleration of the atherosclerotic process Treatment with antioxidants and with inhibitors of the renin-angiotensin system may reverse some of the pathologic vascular changes associated with endothelial dysfunction. PMID- 11727408 TI - Treatment of dyslipidemia in diabetes. AB - Atherosclerosis kills more patients with diabetes than all other causes combined. Treatment must be focused on several targets: glycemic control, bulk reductions of LDL cholesterol, and shifting LDL particle size. Aggressive treatment and reversal of dyslipidemias is a proven prevention for coronary events in patients with type 2 diabetes. Glycemic control with diet, oral hypoglycemic agents, and insulin, when necessary, is often only partially effective in normalizing lipid values in type 2 diabetes. Intensive treatment with lipid-regulating agents, particularly statins, is often necessary to normalize diabetes-associated dyslipidemias. Statins are also the only agents thus far shown in prospective, controlled trials to reduce the risk of coronary events in diabetic patients definitively. PMID- 11727409 TI - The academic alliance for AIDS care and prevention in Africa. PMID- 11727410 TI - What is the latest news on structured interrupted treatment? Should a healthy HIV positive person consider an STI? What are the risks involved and for how long should the meds be interrupted? Is there any chance of developing resistance? PMID- 11727411 TI - AIDS freak-time to give up? PMID- 11727412 TI - After a person has built resistance to a drug and then goes on other drugs for a while, is it possible that the virus loses its resistance to the first drug after not being in their system for awhile, or will the virus be resistant to that drug for the rest of his/her life? PMID- 11727413 TI - My doc wants me to have another genotype panel. The HMO will not pay for this test. Last time, it cost my lover and me more than $900 from a lab in California. Do you know of some less costly providers of genotype tests? $900 is a lot of money! And do you have any information on when (if ever) the FDA is going to approve these tests, so the insurance companies will pay for them? PMID- 11727414 TI - Is there a point at which you would recommend discontinuing treatment due to liver toxicity, short of liver failure? What, for instance, would be the numbers to look for with liver enzyme tests? How high is too high? My doctor has said they are seeing more people in their practice die from liver disorders than from the virus itself nowadays. This has me very concerned since I am co-infected with HCV. PMID- 11727415 TI - Understanding the role of personality in HIV risk behaviors: implications for prevention and treatment. PMID- 11727416 TI - HIV and paradoxical worsening of tuberculosis. PMID- 11727417 TI - Immune recovery uveitis. PMID- 11727418 TI - Implementing a patient management system in an emergency unit and traumatic clinic setting. PMID- 11727419 TI - Excellent project management gives win-win for West Glasgow NHS trust and MDIS. PMID- 11727420 TI - Health Management Strengthening Project. West Bank and the Gaza strip. PMID- 11727421 TI - Recording the final diagnosis. PMID- 11727423 TI - "Setting the record straight". A review of progress in health records services. PMID- 11727422 TI - "Heard it on the grapevine". Tough new monitoring regime for out-patients unveiled as new weapon in the war on NHS waiting. PMID- 11727424 TI - Beneath the veil. Health Records in the United Arab Emirates. PMID- 11727425 TI - Health record services rules & regulations. PMID- 11727426 TI - Standards for medical record services. PMID- 11727427 TI - Running your own IHRIM certificate course. The Trent Branch experience. PMID- 11727428 TI - Pets are good for you, say doctors. PMID- 11727429 TI - Heard it on the grapevine. The Employment Relations Act 1999. PMID- 11727430 TI - Medical negligence and access to medical records. PMID- 11727431 TI - Beneath the veil--health records in the Middle East. AB - The healthcare industry is a fast-growing, dynamic industry within the UAE today. It is a basic necessity that is constantly changing in line with advancing technology, and the need to provide the right service to the right patient at the right time. The MAT programme aims to provide its students with the education necessary to enable them to meet and exceed the needs of these demands. The three cornerstones of this programme will ensure that this is achieved, as with quality, standards and confidentiality for clear signposts to industry relevance, the students cannot fail to reach their potential. PMID- 11727432 TI - Medical records in the governmental hospitals, Gaza Strip, Palestine. PMID- 11727433 TI - Digital archiving: a first for Walsall Hospitals NHS Trust. PMID- 11727434 TI - Bradford Hospitals NHS Trust: the ultimate experience. PMID- 11727436 TI - Project data management: medical records--the way forward. PMID- 11727435 TI - Altnagelvin cracks the codes! PMID- 11727437 TI - Quality assurance. PMID- 11727438 TI - Swindon & Marlborough official launch. PMID- 11727439 TI - Report from the 5th AIDS Impact Conference. HIV prevention at the start of the new millennium. PMID- 11727440 TI - Mental health and quality of life for people with HIV. PMID- 11727441 TI - Conference excerpt: living long-term with HIV. PMID- 11727442 TI - Herbs and drugs: a risky mix. PMID- 11727443 TI - Time to throw away your reading glasses? PMID- 11727444 TI - New tools for taking control of Alzheimer disease. PMID- 11727445 TI - What to do if prostate cancer returns. PMID- 11727446 TI - My atrial fibrillation has returned after five years. Why? What should I do? PMID- 11727447 TI - Why do you recommend swimming for people with osteoporosis even though it is not weight bearing? PMID- 11727448 TI - Health system tackles severity-adjusted profiling. PMID- 11727449 TI - Sacramento hospital boosts outcomes by focusing on high-risk CHF patients. AB - Sacramento's Mercy Heart Institute turns to one-on-one counseling to reduce the risk of hospitalization among congestive heart failure patients. One innovative spin is the use of a high-tech linkage to home called Health Hero. PMID- 11727450 TI - Successful DM efforts make the case for investment in hypertension control. AB - Why? Because data consistently show that only a small percentage of people diagnosed with this chronic condition are adequately controlled. Despite the fact that this is an obvious opportunity for improvement, few organizations have launched DM efforts specifically targeting hypertension, primarily because of the difficulty inherent in predicting ROI. However, DMA reviews two approaches that are getting notable results. PMID- 11727451 TI - Asthma emerges as significant problem for elderly.. AB - Physicians are taught the old adage that when you hear hoof beats, "think horses, not zebras" when diagnosing common symptoms. According to two recent studies by Johns Hopkins Medical Institutions' researchers, physicians aren't thinking "allergy" enough when it comes to their asthmatic patients. PMID- 11727452 TI - Gavestinel produces no benefit for stroke patients, study finds. AB - There was only problem, it didn't work. The wonder drug, gavestinel, failed to produce any significant treatment benefits for patients treated within six hours after experiencing an acute ischemic stroke, according to the recent results of a major clinical trial of the neuroprotectant. The outcome has dimmed the optimism of hospitalists that rapid treatment would dramatically improve patient outcomes. PMID- 11727453 TI - A scary halloween for providers. Hospitals, doctors eye options after CMS issues regs that include rate cuts. PMID- 11727454 TI - Who's minding the store? Survey finds need for more than 100,000 nursing-home workers. PMID- 11727455 TI - Two views of Anthem's IPO. Blues giant sees economies of scale, while critics fear higher premiums, less care. PMID- 11727456 TI - Idaho merger mania? After years of no activity, four of state's 42 hospitals to consolidate. PMID- 11727457 TI - Oh, predictability, where art thou? CMS' Scully seeks stable Medicare funding in an unstable fiscal era. PMID- 11727458 TI - Disproportionate burden. Providers near U.S.-Mexico border face a flood of problems in caring for immigrants. PMID- 11727459 TI - Competition heats up in Chile. Healthcare reform proposals seek increasing role for nation's private sector. PMID- 11727460 TI - Foreign fallout from 9-11. U.S. healthcare firms' small but growing dealings abroad face uncertainty. PMID- 11727461 TI - A means in itself. Ventricular-assist devices gain notice as possible treatment for heart patients. PMID- 11727462 TI - Good, but not good enough. For-profit chains post strong earnings, but Wall Street still disappointed. PMID- 11727463 TI - 'We're all working together.' New York-area hospitals redouble efforts in face of anthrax death. AB - The death of a New York hospital worker from inhalation anthrax last week put new strains on an overburdened public health system. The fact that no source of the bacteria could be found only added to the difficulties. Hospitals along the East Coast battled clinical, logistical and financial problems from the outbreak. PMID- 11727464 TI - Whatever the number, it's big. AHA spreads confusion with differing costs for hospitals' terrorism preparedness. PMID- 11727465 TI - Organized medicine mobilizes. Using wealth of personnel, professional skills to help nation address grim reality. PMID- 11727466 TI - The evaluation of pharmacologic therapy in humans: a brief summary of the drug evaluation process and guidelines for clinical trials as they related to women. AB - Significant progress has been made in including women in clinical and drug evaluation trials. Nonetheless, for most drugs currently on the market, analysis of benefits by sex is not available. At least some of the adverse effects of newer drugs in women could be due to the lack of inclusion in studies from which therapeutic regimens were derived. The data currently available on potential sex differences in pharmacokinetics and pharmacodynamics are also limited by having been obtained from healthy subjects receiving only one medication in studies designed only to detect moderate-to-large (> 30-50%) differences between the sexes. The clinical environment is different: patients consume multiple medications, including over-the-counter medications as well as nutraceuticals and dietary supplements; patients are, on average, older than healthy volunteers or even patients enrolled in investigational studies; and patients are more likely to have multiple diseases. In addition, adequate numbers of women still have not been enrolled in clinical trials for the therapy of many common disorders. The prudent clinician will remember that every time a therapy is initiated for an individual patient, especially a female patient, it is a clinical trial and the outcome is uncertain. PMID- 11727467 TI - Overweight, stature, and socioeconomic status among women--cause or effect: Israel National Women's Health Interview Survey, 1998. AB - OBJECTIVE: To determine if short stature and overweight/obesity are predictors of low socioeconomic status (SES), measured by educational attainment. DESIGN: A cross-sectional study on a random sample carried out through telephone interviews over a period of five months (March through August 1998). SUBJECTS: A total of 1587 Jewish and Arab Israeli women--773 women age 20-44 years, and 814 women age 45-74 years. METHODS: Two cross-sectional telephone surveys were conducted; both times, telephone numbers were drawn from the population registry. Multiple logistic regression analysis was used to determine significant predictors of body mass index (BMI), while controlling for population group, parity, smoking, and physical activity. RESULTS: The level of one's education and one's stature were significantly, independently associated with BMI. Low stature and obesity were indicators of low educational attainment. LIMITATIONS: The subjects of the study reported their own weight and height. The tendency to overestimate one's height and underestimate one's weight places the accuracy of data under question. However, as there is no reason to believe that the biases in the reports would vary with educational status, the results of the study remain valid. CONCLUSIONS: Obesity/overweight is directly related to the level of education and inversely to stature. Together, one's level of education and stature may be determinants of low SES. Therefore, in subpopulations with high rates of obesity and short stature, comprehensive programs should include welfare in addition to dietary measures. PMID- 11727468 TI - Anterior cruciate ligament injuries in the female athlete. AB - With the participation of women in athletics growing rapidly over the last two decades, a disturbing gender-specific pre-disposition has emerged regarding anterior cruciate ligament (ACL) injuries of the knee. Female athletes have a two to eightfold higher incidence of ACL injury than their male counterparts. It is estimated that 38,000 women sustain ACL tears per year. The majority of ACL injuries in female athletes occur through noncontact mechanisms, most often during deceleration activities, such as landing from a jump or cutting. The risk factors for noncontact ACL injuries can be categorized as intrinsic (anatomic and hormonal) and extrinsic (environmental and biomechanical). This article will discuss these risk factors that are thought to contribute to the higher incidence of ACL injuries in women, the development of prevention strategies, and the outcomes of ACL reconstruction in women. PMID- 11727469 TI - Sexual function and quality of life: endpoints and outcomes. AB - Quality of sexual functioning and quality of life (QOL) are implacably interwoven phenomena. Therefore, realistic clinical trials of the efficacy of potential therapeutic agents for sexual dysfunction must include endpoints that reflect both sexual functioning and QOL. A review of design parameters in clinical trials demonstrates that the selection of well-developed psychological outcomes measures to define these endpoints can be an important factor in improving the sensitivity of clinical trials. The inclusion of well-designed and validated outcomes measures increases sensitivity through reductions in measurement of error and the deflection of additional variance associated with within-subjects variability. A brief list of recommended outcomes measures for both sexual functioning and relevant QOL domains is included. The roster of measures is not intended to be comprehensive; however, the instruments cited all have been well-designed, conform to the requirements of clinical trials, and have demonstrated sensitivity to therapeutic effects in previous investigations. PMID- 11727470 TI - Doctrine at the gate: religious restrictions in health care. PMID- 11727471 TI - Red crossroads. PMID- 11727473 TI - Early outbreak alert. PMID- 11727472 TI - Germs and guns. PMID- 11727474 TI - Bleach, gas, or foam? PMID- 11727475 TI - Missing in action. With no one at the wheel, the FDA has been conspicuously voiceless during the health crisis. PMID- 11727476 TI - Medicare in the 21st century. AB - The authors provide an overview of the Medicare program in terms of how the current program operates, the current issues it faces that may shape possible options for reform, and the implications of these features and issues for employment-based health plans. Current issues include adoption of a premium support model, changes in the eligibility age for Medicare benefits, Medigap insurance, benefits covered and customer service. PMID- 11727477 TI - Retiree health care benefits: new rules, new strategies. AB - Two recent court decisions in a retiree health care benefits age discrimination case are likely to bring changes to employer-sponsored retiree health care. After reviewing the cases and how the Equal Employment Opportunity Commission has responded to them, this article discusses employer implications. Although the court decisions add complexity to retiree health plan design, they also provide employers with the valuable opportunity to analyze the match between their retiree health plans and business needs. PMID- 11727479 TI - Disability retirement benefits--workers' compensation setoff--attorney fees and costs. Trujillo v. Cyprus Amax Minerals Company Retirement Plan Committee. PMID- 11727478 TI - A sign of the times. AB - Pressure placed on employers to enhance the bottom line comes from a multitude of changes in the marketplace and has resulted in employers seeking to limit their liabilities and expenses for retiree medical care. To address retiree medical issues, employers have adapted a number of strategies that are presented in this article. These include eliminating or modifying benefits, using a defined contribution approach, capping subsidies and enrolling retirees in Medicare HMOs. PMID- 11727480 TI - Health plan reimbursement--offsets. Harris v. Harvard Pilgrim Health Care, Inc. PMID- 11727481 TI - Administration--health benefits--reimbursement. Reynolds Metals v. Ellis. PMID- 11727482 TI - Canadians less satisfied with hospital care, but more supportive of private alternatives. PMID- 11727483 TI - Use of health services by Ontario children declining. PMID- 11727484 TI - Moving beyond healthcare: the role of healthcare organizations in creating healthy people in healthy communities in a healthy world. PMID- 11727485 TI - Walkerton water and complex adaptive systems. PMID- 11727486 TI - Healthy hospitals: a journey to ISO 14001 certification. PMID- 11727487 TI - Neglect of the inpatient: the hospitalist movement in Canada responds. PMID- 11727488 TI - Hospitalist model: advantages and concerns. PMID- 11727489 TI - Hospitalists: a bandage on a gaping wound. PMID- 11727490 TI - Physician supply in Canada: how much is enough? PMID- 11727491 TI - Nursing shortages: workplace environments are essential to the solution. PMID- 11727492 TI - Healthy workplaces mean more satisfied nurses. PMID- 11727493 TI - E-commerce legislative initiatives in Canada: implications for healthcare institutions. PMID- 11727494 TI - Developments in liposomal gene delivery systems. PMID- 11727495 TI - Papillomavirus-like particle based vaccines: cervical cancer and beyond. AB - Non-infectious human papillomavirus-like particles (VLP), composed of the L1 major capsid protein, are under active development as vaccines to prevent cervical cancer. They would presumably function primarily by generating virion neutralising antibodies against the genital human papillomavirus (HPV) types that are the principal cause of most cervical cancers. Early phase clinical studies indicate that the VLP vaccines are well tolerated and able to consistently induce high titres of virus type-specific neutralising antibodies. Two types of second generation VLP-based subunit vaccines with therapeutic implications, both related and unrelated to papillomavirus infection, are in preclinical development. One type seeks to induce cell-mediated immune responses, especially cytotoxic lymphocytes (CTL), against non-structural papillomavirus proteins, proteins of other viruses, or tumour associated antigens. The target antigen is incorporated into the VLPs as a fusion protein of L1 or the L2 minor capsid protein. In mouse models, this approach has generated potent CTL responses after low dose vaccination in the absence adjuvant. The second type of therapeutic VLP-based vaccine seeks to generate autoantibodies to self-antigens. The display of self polypeptides in the context of the highly ordered array of repetitive elements on the papillomavirus VLP surface abrogates the ability of the humoral immune system to functionally distinguish between foreign and self. High titre and high avidity auto-reactive IgG antibodies have been induced to both soluble (TNF-alpha) and cell surface (CCR5) central self-antigens. Vaccines based on this approach could potentially be effective alternatives to monoclonal antibody (mAb)-based therapies for a variety of disease targets. PMID- 11727496 TI - Block copolymeric biotransport carriers as versatile vehicles for drug delivery. AB - This review describes block copolymer-based systems that are used in drug formulation development. The use of amphiphilic block copolymers to modify pharmacological performance of various classes of drugs attracts more and more attention. This is largely attributable to the high tendency of block copolymer based drug formulations to self-assemble, as well as flexibility of block copolymer chemistry, which allows precise tailoring of the carrier to virtually any chemical entity. Combination of these features allows adjustment of block copolymer-based drug formulations to achieve the most beneficial balance in drug biological interactions with the systems that control its circulation in and removal from the body and its therapeutic activity. The following major aspects are considered: 1) physical properties of formulations and the methods used to adjust these properties towards the highest pharmacological performance of the product; 2) combinatorial methods for optimisation of block copolymer-based formulations; 3) biological response modifying properties of block copolymer based formulations. PMID- 11727497 TI - Genetically engineered antibodies for direct antineoplastic treatment and systematic delivery of various therapeutic agents to cancer cells. AB - Classical antineoplastic therapeutic modalities such as surgery, radiation, and chemotherapy not only fail to cure the great majority of neoplasms, but their employment often leads to severe and debilitating side effects associated with severe neoplasm-related morbidity. Immunotherapy as a fourth modality of anti cancer therapy has already been proven to be quite effective. The astonishing immunophenotypic (IP) heterogeneity of neoplastic cells, the different cytotoxic activity associated with the moiety linked to given monoclonal antibodies (mAb), and mostly the impressive genetic modulation capabilities of cancer cells still remain as yet unsolved difficulties in the present immunotherapy of human neoplasms. The advances in mAb production have revitalised the initial concept of use of cancer cell specific "magic bullets." Antibodies represent new approaches to anti-cancer therapy: they are neoplastic cell-specific and lethal to neoplastically transformed cells via immune effector mechanisms with no toxicity to normal tissues. They are being observed and developed, adhering to the old prayer: "Destroy the diseased tissues, preserve the normal." Strategies for the employment of antibodies include: 1) immune reaction directed destruction of neoplastic cells; 2) interference with the growth and differentiation of malignant cells; 3) antigen epitope directed transport of anti-cancer agents to neoplastic cells; 4) anti-idiotype tumour vaccines; and 5) development of engineered (humanized) mouse mAbs for anticancer therapy. In addition, a variety of agents (e.g. toxins, radionuclides, chemotherapeutic drugs) have been conjugated to mouse and human mAbs for selective delivery to neoplastic cells. PMID- 11727498 TI - Current developments in malaria transmission-blocking vaccines. AB - Malaria is still a leading cause of morbidity and mortality in human populations. Problems, including drug-resistant parasites and insecticide resistant mosquitoes, ensure the continued hold of malaria in the tropics and sub-tropics. Each year around 100 million cases of malaria result in at least 50,000 deaths outside of sub-Saharan Africa; within sub-Saharan Africa itself, malaria causes around one million child deaths per year. New approaches for malaria control are badly needed and much effort has gone to develop malaria vaccines. In addition to giving personal protection, most such vaccines would also tend to reduce the transmission of malaria. One class of vaccine is being developed specifically for this purpose--the malaria transmission-blocking vaccines (TBV). TBVs are based upon antigens expressed on the surface of the sexual and mosquito mid-gut stages of malaria parasites. These antigens are the targets of antibodies induced by vaccination of the host and ingested with the parasites in a mosquito blood meal. The antibodies act by inhibiting the parasite's development within the mosquito itself and they thereby prevent the onward transmission of the parasites. TBVs could contribute to the total interruption of malaria transmission in many locations with relatively low transmission rates, mostly outside sub-Saharan Africa. Under almost all transmission rates, however, TBVs would help reduce malaria incidence and malaria-related morbidity and mortality. Promising recombinant TBV candidate antigens for the two main human malaria parasite species, Plasmodium falciparum and Plasmodium vivax, have been produced and tested in the laboratory; one has undergone early clinical trials. PMID- 11727499 TI - Gene therapy strategies for the treatment of chronic viral hepatitis. AB - Chronic viral hepatitis is a major clinical problem, with over half a billion persons infected worldwide. Current therapies, principally treatment with recombinant IFN-alpha protein, have limited benefit. Recent studies suggest that gene-based expression of IFN-alpha is a possible therapeutic alternative that may improve the effectiveness of treatment. Gene delivery to the liver and consequent IFN-alpha expression therein, has the potential to concentrate the protein at the target organ and provide more continuous exposure to the therapeutic agent. Other potential gene and nucleic acid therapeutics for viral hepatitis are also being investigated. Key to the deployment of these future therapies is a suitable method of gene delivery. Although recombinant viral vector systems, such as adenovirus, are currently the most effective means of gene delivery to the liver, their use presents many concerns. These include immune and inflammatory reactions to the viral vector and possible adverse interactions between the recombinant virus and the pre-existing viral infection. Non-viral gene delivery systems would be a preferred treatment modality. The efficiency of current non-viral systems is not adequate for systemically administered liver gene therapy. However, recent use of membrane permeabilisation techniques has shown that high efficiency non viral gene transfer agents are possible. The future coupling of these improved delivery systems with gene- or nucleic acid-based therapeutics currently in development holds out great promise for new generations of antihepatitis therapies. PMID- 11727500 TI - Fighting infection using immunomodulatory agents. AB - The last decade has seen the emergence of immunomodulators as promising therapeutic agents in infectious diseases. A diverse array of recombinant, synthetic and natural immunomodulatory preparations for prophylaxis and treatment of various infections are available today. Some of these substances, such as granulocyte colony-stimulating factor (G-CSF), interferons, imiquimod and bacterial-derived preparations are already licensed for use in patients. Others including IL-12, various chemokines, synthetic cytosine phosphate-guanosine (CpG) oligodeoxynucleotides and glucans are being investigated extensively in clinical and preclinical studies. Immunomodulatory regimens offer an attractive approach as an adjunct modality for control of microbial diseases in the era of antibiotic resistance. Practical application of the advances in molecular biology, bioinformatics, genomic mining and high-throughput peptide synthesis should foster future discovery and development of novel immunomodulators contingent upon scientific evidence rather than dictates of discursive empiricism. PMID- 11727501 TI - Gene therapy for hypertension: sense and antisense strategies. AB - Gene therapy for hypertension is needed for the next generation of antihypertensive drugs. Current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). Gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). We are currently using two strategies: antisense oligodeoxynucleotides (AS-ODN), an dantisense DNA delivered in viral vectors, to inhibit genes associated with vasoconstrictive properties. It is not necessary to know all the genes involved in hypertension, since many years of experience with drugs show which genes need to be controlled. AS-ODNs are short, single-stranded DNA that can be injected in naked form or in liposomes. AS-ODNs, targeted to AT1 receptors (AT1R), angiotensinogen (AGT), angiotensin converting enzyme (ACE) and beta 1-adrenergic receptors effectively reduce hypertension in rat models (SHR, 2K-1C and cold induced) hypertension. The effects can last up to one month when delivered with liposomes. No side effects or toxic effects have been detected and repeated injections can be given. For the vector, adeno-associated virus (AAV) is used with a construct to include a CMV promoter, antisense DNA to AGT or AT1R and a reporter gene. Results in SHR demonstrate reduction and slowing of hypertension development with a single dose administration. Left ventricular hypertrophy is also reduced by AAV-AS-AGT treatment. Double transgenic mice (human renin plus human AGT) with high angiotensin II (Ang II) causing high blood pressure, treated with AAV-AT1R-AS, show a normalisation of blood pressure for over 6 months with a single injection of vector. We conclude that ODNs will probably be developed first because they can be treated like drugs for the treatment of hypertension with long-term effects. Viral vector delivery needs more engineering to be certain of its safety but one day may be used for a very prolonged control of blood pressure. PMID- 11727502 TI - Gene therapy for paediatric leukaemia. AB - Improvements in the chemotherapeutic and transplant regimens have had a significant impact in improving survival rates for paediatric leukaemia. However, there are still important problems to address including what options are available for patients with chemoresistant disease and what strategies are available to avoid the concerns regarding the toxicity associated with highly cytotoxic treatment regimens. Gene therapy and immunotherapy protocols hold great promise. Using gene transfer of a marker gene, a number of biological issues in the therapy of leukaemia have been addressed. For example, by gene marking autologous bone marrow grafts it has been possible to demonstrate that infused marrow contributes to relapse in acute and chronic myeloid leukaemias. In the allogeneic transplant setting, genetically modified T-cells have proven valuable for the prophylaxis and treatment of viral diseases and may have an important role in preventing or treating disease relapse. Gene transfer is also being used to modify tumour function, enhance immunogenicity, and confer drug-resistance to normal haematopoietic stem cells. With the continued scientific advancements in this field, gene therapy will almost certainly have a major impact on the treatment of paediatric leukaemia in the future. PMID- 11727503 TI - Immunotherapeutic and antitumour potential of thalidomide analogues. AB - The immunomodulatory drug thalidomide has been shown to be clinically useful in a number of conditions including various immunological disorders and cancers. Clinical activity in vivo is attributed to the wide ranging immunological and non immunological properties possessed by this drug; these include anti-TNF-alpha, T cell co-stimulatory, anti-angiogenic activities and also direct antitumour activity. Recently, the design of compounds based on the thalidomide structure has led to the synthesis of analogues with greatly enhanced immunological activity and with similarly decreased toxicity. These derivatives fail into at least two categories; selective cytokine inhibitory drugs (SelCID), which are phosphodiesterase Type 4 (PDE4) inhibitors and immunomodulatory drugs (IMiD), similar to thalidomide which act via unknown mechanism(s). These compounds are in the process of being characterised in laboratory studies and are also now being assessed in Phase I and Phase I/II clinical studies. In this review we will highlight the properties of these two novel classes of compound in terms of their effects on both immunological and non-immunological systems in vitro. We will also describe how these studies are enabling the characterisation and development of these compounds into clinically relevant drugs in widely varying diseases. To this end we will describe the various clinical studies of lead compounds that are in progress and speculate as to the potential and future development of these exciting compounds. PMID- 11727504 TI - Interference with the urokinase plasminogen activator system: a promising therapy concept for solid tumours. AB - There is abundant evidence that the plasminogen activator (PA) system with its key components uPA (urokinase-type plasminogen activator), its cell surface receptor uPA-R (CD87) and its inhibitor PAI-1 plays a key role in tumour invasion and metastasis. Elevated levels of these factors in tumour tissue are associated with tumour aggressiveness and poor patient outcome. Animal models suggest that the PA system is not essential for fertility or survival under physiological conditions. Thus, it seems well suited as a therapeutic target for patients with solid malignant tumours. Novel therapy concepts targeting the uPA system are currently being explored. A variety of different synthetic uPA inhibitor classes have been developed over the last decades. First generation inhibitors displayed a low uPA inhibitory potency combined with broad specificity. More recently, structure based design, x-ray crystallographic screening or NMR based screening have revealed a large number of new, potent and selective uPA-inhibitors. A few modern compounds have shown promising results in preclinical testing and are now ready for Phase I clinical studies. Other therapeutic strategies such as antagonists of uPA/uPA-R interaction or gene therapeutic approaches to suppress the uPA-system are still being evaluated in in vitro and in vivo models. For clinical application, a combination therapy targeting more than one of the interacting proteolytic pathways may be required for effective antiproteolytic therapy. In addition, antiproteolytic agents may provide additive or synergistic treatment benefits if used in combination together with conventional therapeutics, in particular in those solid tumours for which potent conventional regimens already exist. PMID- 11727505 TI - Recombinant immunotoxins for cancer therapy. AB - Recombinant immunotoxins consist of Fv regions of tumour-selective antibodies fused to toxins found in bacteria, plants or fungi. These toxins must be modified to remove normal-tissue binding sites but to retain all other functions of cytotoxicity. The recombinant antibody fragments target the modified toxin to cancer cells which are killed, either by direct inhibition of protein synthesis, or by concomitant induction of apoptosis. Cells that are not recognised by the antibody fragment because they do not carry the tumour antigen, are spared. Many factors influence the in vivo antitumour activity of recombinant immunotoxins. Among them are considerations of which types of cancer may be the best targets for immunotoxin therapy as well as tumour specificity of the antigen that is targeted by the recombinant antibody. Other relevant issues are the affinity of immunotoxins and their ability to enter and penetrate into tissues and tumours, which in turn is dependent on the size of the protein. A great deal of protein engineering is required to stabilise the recombinant antibody moiety of immunotoxins, since stability of the molecules is crucial for good clinical efficacy. Excellent activity and specificity can be observed for many recombinant immunotoxins in in vitro assays using cultured cancer cells as well as in animal tumour models. Ongoing clinical trials provide examples where the promising preclinical data correlate with successful results in experimental cancer therapy. PMID- 11727506 TI - VEGF antagonists. AB - The majority of cancer have an absolute requirement for angiogenesis, the process by which new blood vessels are formed. The most potent angiogenic cytokine is vascular endothelial growth factor (VEGF) and there has been substantial research into the development of VEGF/VEGF receptor (VEGFR) antagonists. To date these strategies have included gene therapy techniques that deliver antisense oligonucleotides, soluble VEGFRs that function in a dominant negative fashion and ribozymes. Additional strategies have included the development of receptor tyrosine kinase (RTK) inhibitors and monoclonal antibodies (mAbs) directed against VEGF or the signalling receptor. The most promising agents appear to be the monoclonal anti-VEGF antibodies and the RTK inhibitors as these have demonstrated broad spectrum antitumour activity in vivo and single agent activity in early phase clinical trials in patients with advanced pre-treated breast and colorectal carcinoma and Kaposi's sarcoma. The RTK inhibitors are of particular interest as they can be administered by mouth. Collation of the early clinical trial data suggests that VEGF antagonists are largely well-tolerated but may be associated with vascular toxicities such as haemorrhage and thromboembolic events. Combination studies of chemotherapy and VEGF antagonists are underway but the benefit of these regimens will need to be established in adequately powered Phase III studies. Potentially these agents may play a role in the treatment of both early (adjuvant) and advanced cancer. The efficacy of the drugs will be explored in a number of non-malignant conditions including rheumatoid arthritis (RA), psoriasis, diabetic retinopathy and possibly as non-steroidal contraceptives but the overall clinical development of these agents can only be optimised if appropriate biological end points are identified and incorporated into clinical trials. PMID- 11727507 TI - IMC-C225, an anti-epidermal growth factor receptor monoclonal antibody, for treatment of head and neck cancer. AB - Squamous cell carcinoma (SCC) of the head and neck (H&N) remains a clinical challenge due to its high rate of locoregional disease recurrence. The importance of the epidermal growth factor receptor (EGFR) in the development and progression of many solid tumours (including SCC of the H&N) is well understood; increased expression is associated with enhanced tumour invasion, resistance to chemotherapy and decreased patient survival. Several approaches have been developed to achieve EGFR blockade as an anticancer treatment strategy, including an anti-EGFR monoclonal antibody (mAb), IMC-C225, which competitively binds to the extracellular receptor site to prevent binding by natural EGFR ligands (EGF and TGF-alpha). Preclinical studies evaluating this chimeric mAb in human cancer cell lines in vitro and human tumour xenografts in vivo have demonstrated its potent antitumour activity. The clinical efficacy of IMC-C225 appears to involve multiple anticancer mechanisms, including inhibition of cell cycle progression, induction of apoptosis, anti-angiogenesis, inhibition of metastasis and its ability to enhance the response to chemotherapy and radiation therapy. Phase I studies of IMC-C225 combined with chemotherapy or radiation for SCC of the H&N demonstrate excellent response rates in patients with recurrent or refractory disease. Phase II and III trials examining the efficacy and safety of these combinations are currently underway. To date, IMC-C225 has been well-tolerated, with skin rashes and allergic reactions being the most clinically important adverse events reported. IMC-C225 displays dose-dependent elimination characteristics and a half-life of approximately 7 days. Current recommendations for dosing include a 400 mg/m2 loading dose, followed by weekly infusions of 250 mg/m2. PMID- 11727508 TI - The clinical potential of novel erythropoiesis stimulating protein. AB - Novel erythropoiesis stimulating protein (NESP) is a supersialylated analogue of endogenous erythropoietin or recombinant human erythropoietin (rhuEPO). It contains a total of five N-linked consensus carbohydrate binding sites in the native protein. NESP has a higher molecular weight due to an increased content of carbohydrates, which, however, has no meaningful influence on the binding to and activation of the erythropoietin receptor. The major difference in comparison to rhuEPO is the up to threefold increase of the terminal half-life of NESP, which allows for less frequent dosing of NESP. Several clinical studies have shown that NESP is as safe and efficient as rhuEPO in correcting renal anaemia and in the maintenance therapy of renal anaemia. PMID- 11727509 TI - Complementary and alternative medicine in cancer: a National Cancer Institute perspective. PMID- 11727510 TI - Virokines: novel immunomodulatory agents. AB - Since the discovery of virokines in the 1980s, much time and research has been dedicated to exploring their potential use as therapeutic agents. Simply put, virokines are virally encoded proteins that are secreted from the infected host cell. Most of these proteins possess the ability to modulate different aspects of the host immune system, to better maintain a suitable habitat for viral replication. These proteins are often highly homologous to host immune proteins but are often smaller and more powerful. Examples of virokines include viral secreted proteins that: block components of the complement system, act as serine protease inhibitors, function as chemokine and cytokine agonists or antagonists and contribute to cell proliferation. Many of these proteins are currently being investigated for use as novel therapeutic immunomodulators to manage immune disorders, inflammation after trauma, graft rejection and autoimmune diseases. PMID- 11727511 TI - Anticytokine gene therapy of autoimmune diseases. AB - Viral and nonviral gene therapy vectors have been successfully employed to deliver inflammatory cytokine inhibitors (anticytokines), or anti-inflammatory cytokines, such as transforming growth factor beta-1 (TGF-beta 1), which protect against experimental autoimmune diseases. These vectors carry the relevant genes into a variety of tissues, for either localised or systemic release of the encoded protein. Administration of cDNA encoding soluble IFN-gamma receptor (IFN gamma R)/IgG-Fc fusion proteins, soluble TNF-alpha receptors, or IL-1 receptor antagonist (IL-1ra), protects against either lupus, various forms of arthritis, autoimmune diabetes, or other autoimmune diseases. These inhibitors, unlike many cytokines, have little or no toxic potential. Similarly, TGF-beta 1 gene therapy protects against numerous forms of autoimmunity, though its administration entails more risk than anticytokine therapy. We have relied on the injection of naked plasmid DNA into skeletal muscle, with or without enhancement of gene transfer by in vivo electroporation. Expression plasmids offer interesting advantages over viral vectors, since they are simple to produce, non-immunogenic and nonpathogenic. They can be repeatedly administered and after each treatment the encoded proteins are produced for relatively long periods, ranging from weeks to months. Moreover, soluble receptors which block cytokine action, encoded by gene therapy vectors, can be constructed from non-immunogenic self elements that are unlikely to be neutralised by the host immune response (unlike monoclonal antibodies [mAbs]), allowing long-term gene therapy of chronic inflammatory disorders. PMID- 11727512 TI - Human leukocyte differentiation antigens as therapeutic targets: the CD molecules and CD antibodies. AB - The cell membrane presents an attractive target in a number of different disease situations. Most obviously, malignant cells may be killed by damaging their cell membranes. There are also more subtle, though effective, ways of rendering cells harmless by engaging proteins at the cell surface. The cells of the immune system may be targeted, for example to stop a damaging immune reaction, such as acute inflammation or rejection of a transplanted organ. If we are to make the best use of the opportunities to modulate disease by targeting the cell membrane, we need a detailed understanding of the many proteins, glycoproteins and glycolipids that are attached to or inserted in the cell membrane. The CD (cluster of differentiation) Workshops, more properly known as the HLDA (Human Leukocyte Differentiation Antigens) Workshops have, since 1982, focussed on the study of the membrane molecules of leukocytes, including the major cells of the immune system and malignant cells derived from them. The scope has extended to molecules on endothelium which are important in interaction with leukocytes. Many of the molecules characterised as leukocyte antigens are also expressed on other tissue. The approaches developed by the HLDA Workshops are useful in the study of the molecular composition and function of cells of other organ systems. Some of the antibodies produced in order to study the CD molecules have proved useful as therapeutic agents. This review describes the CD system, how it has developed and what it means and introduces the field of therapy based on antibodies against CD or similar molecules. The author is responsible for organising the next (8th) HLDA Workshop and invites readers to suggest ways in which the therapeutic relevance of the Workshop may be enhanced. PMID- 11727513 TI - Vaccines against meningococcal disease: current and future technologies. AB - Development of the meningococcal serogroup C conjugate vaccine and its national implementation in the UK has been a major breakthrough in the prevention of meningococcal disease. New technologies are increasing the likelihood that research towards a vaccine against group B meningococcus will be successful. This review covers the recent development of vaccines against meningococcal disease and examines future vaccine candidates. The development of meningococcal polysaccharide vaccines was based on the virulence of the bacterial capsule components. The immunogenicity of these vaccines has been improved by covalent linkage to proteins in the new meningococcal C conjugate vaccines. However, the most promising developments for serogroup B disease have stemmed from other virulence determinants such as outer membrane proteins (OMPs) and lipopolysaccharides (LPS). New genome sequencing technology promises a way forward to developing a broadly cross-protective vaccine for this important pathogen. PMID- 11727514 TI - Development of a cytomegalovirus vaccine: lessons from recent clinical trials. AB - Cytomegalovirus-caused diseases are preventable. We believe that both neutralising antibodies and cell-mediated immunity are necessary for prevention. Of the CMV proteins, gB and pp65 are the minimum requirements in a vaccine to induced neutralising antibodies and cytotoxic T-lymphocyte (CTL) responses. Immunisation with additional proteins, e.g., gH, gN for neutralising antibodies and IE1exon 4 and pp150 for CTL responses, would strengthen protective immune responses. Approaches to development of a safe and effective cytomegalovirus (CMV) vaccine for the prevention of CMV diseases include: a) a live attenuated vaccine (Towne strain); b) recombinant constructs of the attenuated Towne and the virulent Toledo CMV strains; c) subunit glycoprotein B (gB) adjuvanted with MF59 to induce neutralising antibodies; d) phosphoprotein 65 (pp65) peptide-based vaccines to induce (CTL) for use in therapeutic vaccination; e) canarypox-CMV recombinants, e.g., ALVAC-CMV(gB) and ALVAC-CMV (pp65) to induce neutralising antibodies and CTL responses, respectively; f) DNA plasmids containing the genes for gB and pp65; g) dense bodies containing the key antigens. The attenuated Towne strain, gB/MF59, ALVAC-CMV(gB) and ALVAC-CMV(pp65) approaches have already been tested in clinical trials. The Towne vaccine induced neutralising antibodies and cell-mediated immunity (including CTLs) mitigated CMV disease in seronegative renal transplant recipients and protected against a low-dose virulent CMV challenge in normal volunteers but did not prevent infection in mothers of children excreting CMV. Immunisation with gB/MF59 resulted in high levels of neutralising antibodies in seronegative subjects. ALVAC-CMV(gB) did not induce neutralising antibodies but primed the immune system to a Towne strain challenge, while ALVAC-CMV(pp65) induced long-lasting CTL responses in all originally seronegative volunteers, with CTL precursor frequency similar to naturally seropositive individuals. These results suggest that CMV diseases can be prevented or attenuated and that a vaccine combining ALVAC-CMV(pp65) with gB/MF59 may induce sufficient CTLs and neutralising antibodies to protect against CMV diseases. Meanwhile, other approaches such as DNA peptide and dense body vaccines, should enter Phase I trials. All candidate vaccines will have to demonstrate that immunogenicity provides protection. Combined vaccines containing canarypox (ALVAC) vectors to express CMV-pp65 to induce CTLs and of subunit gB, given together with an appropriate adjuvant to induce neutralising antibodies, should be tested in a target population for the prevention of CMV infection and disease. PMID- 11727515 TI - Ongoing trials of immune-based therapies for HIV infection in adults. AB - Highly active antiretroviral therapy (HAART) can significantly alter the clinical course of patients infected with HIV. Unfortunately, effective lifelong HAART may not be a practical or achievable goal because of toxicities, cost, development of viral resistance and patient compliance issues. Immune-based therapies (IBTs) that target the host immune system may serve as rational additions to our current antiretroviral strategies. Investigations into IL-2 have culminated in two large Phase III clinical trials. Multiple therapeutic vaccine candidates are in various phases of investigation. In addition, gene therapy has been proposed as a potential treatment for HIV and Phase I trials are ongoing. Although IBTs are being investigated on many fronts, they remain difficult to study due to a lack of validated surrogate end points. PMID- 11727516 TI - Prospects for IL-16 in the treatment of AIDS. AB - IL-16 is a multi-functional cytokine that uses CD4 as a receptor to signal diverse biological activities by target cells including T-lymphocytes, monocytes and eosinophils. IL-16 has been shown to repress HIV-1 infection in lymphocytes and monocytic cells and it is active against both laboratory and naturally acquired virus isolates. In lymphocytes, the repressive effect of IL-16 occurs at the level of virus transcription, while it appears to inhibit viral entry in monocytic cells. Clinical studies comparing serum IL-16 levels with the state of HIV-1 disease suggest that this cytokine is a functionally significant endogenous antiviral factor. The antiviral activity of IL-16 may be of therapeutic benefit in HIV/AIDS but its greatest potential is for immune reconstitution. Stimulation of CD4+ T-cells with IL-16 primes cells to respond to IL-2, by upregulating the expression of IL-2 receptor p75 (CD25). Co-treatment of peripheral blood mononuclear cells (PBMC) with IL-16 plus IL-2 (or IL-15) in vitro selectively expands the population of CD4+ T-cells. Clinical trials of recombinant IL-2 have already shown promise in HIV/AIDS. In combination with IL-16, the beneficial effects of IL-2 may be augmented and specifically targeted to CD4+ T-cells. Thus, IL-16 shows considerable promise as an agent for the biological therapy of HIV/AIDS. PMID- 11727517 TI - Interleukin-5: a novel target for asthma therapy. AB - Eosinophilic airway inflammation is the main histologic correlate of airway hyper responsiveness (AHR) and tissue injury in the pathogenesis of bronchial asthma. There is strong evidence for a central role of CD4+ T-cells secreting pro allergic Th2-cytokines, such as IL-4 and IL-5, in the induction of airway eosinophilia and AHR. IL-5 appears to be one of the main pro-inflammatory mediators among a growing number of cytokines and chemokines that induce, regulate and sustain eosinophilic airway inflammation. Animal studies provide confirmatory evidence for the important role of IL-5 in the induction and maintenance of eosinophilic airway infiltration leading to altered airway function. Interfering with the action of IL-5 represents one of the new immunomodulatory therapeutic strategies in the treatment of bronchial asthma. Compared to established immunosuppressive agents like steroids, a major advantage of this strategy is the specificity of reducing eosinophilic inflammation, thus possibly acting nearly without side effects. There are several possible ways to inhibit the effects of IL-5 including alteration of the signalling pathway in the IL-5 producing cell by inhibition or modification of transcription factors or the use of antisense oligonucleotides and blocking of the IL-5 protein itself by monoclonal antibodies, soluble IL-5 receptor or antagonists of the IL-5 receptor expressed on the surface of eosinophils. Although preliminary data from the first clinical trials gave rise to skepticism about the efficacy of anti-IL-5 treatment regarding the improvement of lung function of asthmatic patients, further studies with a better defined profile of the target population may provide encouraging results, allowing the introduction of this truly new therapeutic concept. PMID- 11727518 TI - Immunotherapeutic approaches to inflammatory bowel diseases. AB - For a long time corticosteroids, aminosalicylic acid preparations and antibiotics have represented the principal approaches in evidence-based drug therapy for chronic inflammatory bowel diseases (IBD), e.g., Crohn's disease (CD) and ulcerative colitis (UC), and are able to suppress disease activity in most cases. However, there are cases that do not respond to conventional drug therapy or remain dependent on high doses of steroids associated with severe side effects in the long run. It is generally accepted now that IBD has an immunological basis and results from a hyperresponsive state of the intestinal immune system. Although the primary etiological defect respectively immunogenic agent still remains to be identified, substantial progress has been made in our understanding to regulatory mechanisms of the intestinal immune system and their alterations in IBD at the molecular level. Due to the concurrent advent of biotechnological processes it has been possible to utilise these insights for the development of novel immunomodulatory therapeutic strategies ranging from recombinant cytokines and blocking antibodies to oligonucleotide antisense strategies and gene therapeutic approaches. This review will present the current status of the development of these novel immunomodulatory therapeutic strategies in IBD and the status of their use in clinical practice. For a better understanding, it will be necessary to address the recent advances in the elucidation of pathogenetic mechanisms of IBD from studies in human specimen and experimental colitis models that have provided the basis for these novel therapeutic approaches. PMID- 11727519 TI - Neuroprotective possibilities for Huntington's disease. AB - Huntington's disease (HD) is a devastating genetic disorder. Despite the absence of effective therapy, there has been an explosion in interest for developing treatment strategies aimed at lessening or preventing the neuronal death that occurs in this disease. In large part, the renewed interest in neuroprotective strategies has been spurred by our increasing understanding of the genetic and molecular events that drive the underlying neuropathology of HD. This escalating understanding of the biological underpinnings of HD is derived from several convergent sources represented by investigators with clinical, genetic, molecular, physiological and neurobehavioural backgrounds. The diversity of data being generated has, in turn, produced a unique time in HD research where an impressive number of potential therapeutics are coming to the forefront. This review outlines several of these possibilities including the use of intracerebrally delivered neurotrophic factors, pharmacologically altering cellular energy production, the use of antiglutamatergic drugs, the use of caspase inhibitors and inhibitors of protein aggregation. This review also touches on the interesting possibility of whether or not the neurodegeneration in HD is at least partially reversible in nature. All of these possibilities are highlighted in the context that HD is a neurodegenerative disorder in which genetic detection provides a clear and unequivocal opportunity for neuroprotection. PMID- 11727520 TI - In situ gene therapy for prostate cancer: immunomodulatory approaches. AB - The development of effective treatments for prostate cancer is thwarted by the natural history of the disease. The biological and clinical potential of most individual cancers is uncertain. In many cases the disease will not progress to clinical significance but experimental and clinical studies indicate that prostate cancer can and may metastasis early in the course of the disease from relatively small foci (i.e., not necessarily the largest or index cancer). Localised prostate cancer is potentially curable with localised therapies (radical prostatectomy or irradiation therapy). However, there are no curative therapies for metastatic prostate cancer. Gene therapy, especially those approaches with an immunomodulatory component, may provide additional therapeutic options with the potential to affect both localised and systemic disease. We have pioneered the development and application of in situ gene therapy protocols using adenoviral vectors to transduce specific genes that generate cytotoxic activity and/or a systemic antitumour immune response. In addition we have completed initial studies that demonstrate the therapeutic potential of adenoviral vector mediated gene modified cell-based vaccines. Our review discusses preclinical studies focused on the development of immunostimulatory in situ gene therapy approaches that hopefully will provide novel and effective treatments for localised and metastatic prostate cancer. PMID- 11727521 TI - Biological and clinical developments in melanoma vaccines. AB - The identification of antigens recognised on human tumours by autologous T lymphocytes has opened the way for vaccination strategies involving defined tumour antigens. These vaccinations are therapeutic, i.e. they involve patients with detectable disease. Tumour regressions have been observed in a minority of melanoma patients in Phase I/II trials. Some of these regressions have been complete and long lasting. Improving the efficacy of therapeutic vaccines will critically depend on their capacity to trigger a robust immune response, on the development of appropriate methods to monitor these antitumour immune responses to vaccination and on a better understanding of the mechanisms used by tumours to escape immune attack. Finally, the initiation of large randomised Phase III trials will determine the impact of these vaccines on melanoma treatment. PMID- 11727522 TI - T-cell-directed cancer vaccines: mechanisms of immune escape and immune tolerance. AB - Recent clinical trials using vaccines directed toward tumour-associated antigens (TA) have shown the increasing capacity of vaccines to cause immunologic responses. In fact, strongly reactive TA-specific cytolytic T-lymphocytes and tumour-infiltrating lymphocytes (TIL) can be identified and expanded ex vivo from patients with metastatic melanoma vaccinated with melanoma-associated antigens. Paradoxically, this strong immunological response does not correlate with clinical tumour regression. Proposed mechanisms responsible for this glaring inconsistency are numerous and varied; systemic immunosuppressive as well as local mechanistic factors are implicated. In this review we will critically evaluate the possible mechanisms that allow tumours to escape immune destruction and be tolerated by the immune system. In addition, strategies that may allow further insight into the biology of tumour rejection are discussed, in the hope of deepening the understanding of this phenomenon and enhancing its therapeutic potential. PMID- 11727523 TI - Oncolytic virotherapy for cancer with the adenovirus dl1520 (Onyx-015): results of phase I and II trials. AB - Replication-selective viral agents hold promise as a novel cancer treatment platform (virotherapy). dl1520 (Onyx-015, now CI-1042, Pfizer Corp., Groton, CT, USA), an E1B-55kD gene-deleted adenovirus, was the first such genetically engineered agent to be tested in humans. Over 250 cancer patients have now been treated on approximately ten clinical trials (phase I-III). The virus was generally well tolerated at doses of up to 2 x 10(12) particles by intratumoural, intraperitoneal, hepatic arterial and i.v. administration; no maximally-tolerated doses were identified following intra-vascular administration. Viral replication was tumour-selective and was documented after administration by all routes; however, viral replication was variable depending on tumour histology. Single agent efficacy has been relatively limited to date (0-14% local tumour regression rates). In combination with chemotherapy, however, encouraging antitumoural activity has been demonstrated. These clinical research results demonstrate the potential of this novel treatment platform, as well as the hurdles to be overcome. Novel replication-selective agents with improved potency are being developed. PMID- 11727524 TI - HSPPC-96: a personalised cancer vaccine. AB - HSPPC-96 is a protein peptide complex consisting of a 96 kDa heat shock protein (Hsp), gp96, and an array of gp96-associated cellular peptides. Immunisation with HSPPC-96 induces T-cell specific immunity against these peptides; gp96 is not immunogenic per se. The non-covalent binding of gp96 to peptides is neither selective nor restricted to cell types. Thus, the collection of gp96-associated peptides represents the antigenic peptide pool of a cell; this is the basis of the peptide-specific immunogenicity elicited by HSPPC-96. Since the repertoire of antigenic peptides in the cell is different between normal and tumour cells and is also distinct among tumours of the same histology due to the randomness of tumour-specific mutations, a purified sample of HSPPC-96 can only effectively immunise against the tumour from which it originates. The immunisation effect is, therefore, specific to both the individual patient and the tumour type. This true custom-based vaccine has been shown to induce protective T-cell immunity against a variety of tumours in animal models. The major mechanism of action is thought to be the ability of HSPPC-96 to prime peptide-specific MHC class I-restricted CD8+ T-cells by interacting with antigen-presenting cells in a receptor-dependent manner. HSPPC-96 is now being widely tested against human malignancies for which effective therapies do not exist. The major challenges of the human studies are immunological monitoring and the determination of optimal dosing, scheduling and route of administration. HSPPC-96 has been well tolerated at all dose levels. Clinical applications are being addressed by numerous studies including an international, multi-centre, double-blind Phase III trial for the treatment of stage III renal cell carcinoma in the adjuvant setting. PMID- 11727525 TI - Keystone Symposia: chemokines and chemokine receptors. 16-21st February 2001, Taos, NM, USA. AB - Every other year the Keystone Symposia organizes a meeting to discuss the state of the art in chemokine and chemokine receptor research. The focus of the meeting in the past has included the structural and functional identification of chemokines and their receptors. However, this year there was heavy emphasis on the role of chemokines on normal immune function and disease pathogenesis. A number of exciting results were presented and discussed, for example the role of chemokines and chemokine receptors in development and progression of tumours, induction and progression of auto-immunity, development of atherosclerosis and the resolution of infectious disease. The last session of the meeting was devoted to discussion of chemokine and chemokine receptor antagonists currently in preclinical development or Phase I clinical trials. PMID- 11727526 TI - Surrogate markers of response to cancer immunotherapy. AB - Clinically effective cancer immunotherapy has been sought for more than 100 years and has been recently applied most successfully in strategies that passively deliver immune effectors such as monoclonal antibodies (anti-CD20 for lymphoma and anti-HER2/neu for breast cancer), donor lymphocyte infusions in chronic myelongenous leukemia and non-myeloablative allogeneic peripheral blood progenitor transplants for renal cell carcinoma. There is mounting enthusiasm for strategies employing active stimulation of antitumour immune responses. These include vaccines based on tumour antigen proteins and peptides, autologous, allogeneic or gene-modified tumour cells, dendritic cells and antigen-encoding viral vector constructs. Indeed, randomised Phase III clinical trials of autologous tumour cell vaccines for colorectal cancer demonstrated an improvement in disease free survival and a trend toward improved overall survival [1]. Despite these preliminary successes, it is clear that the many strategies under development cannot all be evaluated for survival benefit in large clinical trials that require many years, patients and resources to complete. This highlights the need to develop intermediate markers to help prioritise which agents to test in prospective randomised Phase III trials. PMID- 11727527 TI - Prodrug based optimal drug delivery via membrane transporter/receptor. AB - The carrier-mediated absorption of drugs and prodrugs across epithelial and endothelial barriers is emerging as a novel trend in biotherapeutics. This review examines the important advances in this field in the past decade. The feasibility of drug absorption of the parent drug or the appropriately modified prodrug via these transporters is discussed in detail. Several successful examples of synthesis of prodrugs recognised by the targeted transporters are described. The applicability of this approach in translocating drugs across the almost impenetrable blood-brain barrier (BBB) has also been examined. PMID- 11727528 TI - Alphavirus vectors: development and potential therapeutic applications. AB - Alphaviruses are RNA enveloped viruses that are proving their value as expression vectors. They are particularly well-suited for this role as they are easily and quickly engineered and can be used to produce high levels of proteins of interest. A promising and important use is as vaccines against disease-causing agents such as HIV. The three alphaviruses now serving as vectors are Sindbis virus, Semliki Forest virus (SFV) and Venezuelan equine encephalitis (VEE) virus. Sindbis virus and SFV are well-known models for studies in molecular and cell biology; VEE virus is a human pathogen and had received some previous notoriety as a potential biological weapon. It is now becoming a potentially valuable vaccine vector. All three viruses are being tested as vaccines but, at present, only Sindbis virus and SFV have been considered for other uses. Sindbis virus vectors have been developed to screen libraries for the identification of new proteins and to devise sensitive assays to detect viruses more difficult to grow in culture. Both Sindbis virus and SFV vectors are serving as tools for fundamental studies in biology, examples include development in insects and analysis of protein functions in neuronal cells. In this article the replication strategy of alphaviruses and the different ways they can be engineered to serve as expression vectors is described. This provides an introduction to the ways these vectors have been used and illustrates the promise these vectors offer. PMID- 11727529 TI - Genetically modified tumour vaccines: an obstacle race to break host tolerance to cancer. AB - The development of genetically modified tumour vaccines (GMTV) has been prompted by a better understanding of antitumour immune responses and genetic engineering technologies, as well as the identification of numerous tumour antigens (TA) in several malignancies which occasionally induce spontaneous tumour regressions. Cellular vaccines are based on autologous or allogeneic tumour cells genetically engineered to secrete different cytokines, co-stimulatory molecules, or allogeneic HLA molecules in order to provide a strong stimulatory signal together with the presented TA. Another promising approach that is targeted towards breaking immune tolerance to TA, exploits dendritic cells (DC) loaded or genetically modified with TA (and sometimes cytokines). Effective nonviral and viral gene delivery systems have been constructed including a third generation of adenoviral, lentiviral and hybrid vectors. Studies in mice demonstrated that therapeutic, curative immune responses might be elicited by GMTV. Promising results from animal studies are rarely seen in human trials. Several reasons, such as numerous escape mechanisms of slowly evolving spontaneous tumours and immune incompetence of advanced patients, are major concerns. Improved monitoring of immune responses to GMTV is essential to distinguish between responders and non-responders in order to tailor immune therapy strategy to the individual patient. PMID- 11727530 TI - In utero stem cell transplantation: two steps forward but one step back? AB - The concept of in utero stem cell transplantation has provided hope that at least some of the genetic disorders which can be diagnosed prenatally can be treated before the pathological sequelae become manifested. Our increasing knowledge of haematopoietic stem cell biology has provided further possibilities for optimising this transplant procedure. However, the relatively poor success rate achieved using this procedure has prompted some to suggest that there may be fetal rejection of donor cells via an immune mechanism. However, the prevailing evidence from both animal models and from human cases, suggest that there may be other mechanisms which are preventing successful engraftment in some disorders and these must be addressed if the procedure is to receive further attention. The ability to achieve sustained engraftment after in utero transplantation will have relevance for those seeking to use gene therapy as an alternative therapeutic stratagem. PMID- 11727531 TI - Optimisation of DNA vaccines for the prophylaxis and modulation of herpes simplex virus infections. AB - Herpes simplex virus (HSV) lacks an effective vaccine. Despite its prevalence and importance HSV infection is not controlled with an acceptable vaccine. Perhaps the best candidate and so far untested approach is the use of plasmid DNA encoding viral proteins. Immunomodulators are also holding some hope as a potential therapeutic. In this review various DNA vaccine approaches used in animal model systems to prevent HSV infections are discussed. Judgements are made as to which of these may prove effective for prophylactic or therapeutic vaccines in humans. PMID- 11727532 TI - Potential of anticytokine therapies in central nervous system ischaemia. AB - Central nervous system (CNS) ischaemia is associated with an acute inflammatory response which appears to potentiate CNS injury, especially following reperfusion. This response includes the release of inflammatory mediators called cytokines including IL-1 and TNF-alpha, which triggers the production of additional cytokines including IL-6 and activates leukocytes which infiltrate into the CNS. Increased expression of cytokines has been demonstrated to occur in the first few hours after CNS ischaemia. Preliminary clinical studies suggest that plasma levels of IL-6 are correlated with functional recovery while brain levels of cytokines have been demonstrated to increase following experimental ischaemia. Although there are no current clinical 'anti-cytokine' treatment studies for stroke, experimental studies modulating IL-1 and TNF-alpha have shown neuroprotection. PMID- 11727533 TI - Gene therapy for high grade gliomas. AB - High grade gliomas in adults are devastating diseases, with very poor survival despite their lack of distant metastases. Local treatments, such as surgical resection and stereotactic radiosurgery, have been most successful, whereas systemic therapy (for example, chemotherapy and immunotherapy) have been rather disappointing. Several gene therapy systems have been successful in controlling or eradicating these tumours in animal models and are now being tested as a logical addition to current clinical management. This review describes the gene therapy clinical protocols that have been completed or that are ongoing for human gliomas. These include the prodrug activating system, herpes simplex thymidine kinase (HSVtk)/ganciclovir (GCV), utilising either retrovirus vector producer cells or adenovirus vectors; adenovirus mediated p53 gene transfer; adenovirus mediated IFN-beta gene transfer and oncolytic herpes virus and adenovirus vectors. To date, all of the clinical studies have used direct injection of the vector into the glioma. The Phase I clinical studies have demonstrated low to moderate toxicity and variable levels of gene transfer and in some cases anti tumour effect. Future directions will rely upon improvements in gene delivery as well as gene therapies and combinations of gene therapy with other treatment modalities. PMID- 11727534 TI - General aspects of anti-angiogenesis and cancer therapy. AB - Angiogenesis is the outgrowth of new vessels from pre-existing ones. Tumour growth and metastasis is dependent on angiogenesis and many stimulatory and inhibitory factors have been described which play an active role in this process. Inhibition of tumour neovasculature may be one strategy to inhibit tumour growth. Naturally occurring inhibitors of angiogenesis have been discovered and synthetic agents have been designed. Many of these inhibitors are currently being evaluated in clinical trials for the treatment of cancer. This review discusses the mechanism of action of these anti-angiogenics as well as a description of the clinical trials in which they are being evaluated. PMID- 11727535 TI - T-cell-directed cancer vaccines: the melanoma model. AB - Significant advances in the understanding of the molecular basis for tumour/host interactions in humans have occurred in the last decade through studying patients with metastatic melanoma. This disease is characterised by its tendency to be modulated by immunologic factors. Furthermore, immunologic manipulation of the host with various systemic agents, in particular IL-2, frequently affects this natural phenomenon and can lead to complete rejection of cancer. By studying the cellular immunology occurring in patients undergoing immunotherapy, several tumour antigens (TA) and their epitopes recognised by human leukocyte antigen (HLA) class I-restricted cytotoxic T-lymphocytes (CTL) have been identified. Most of these TA are non-mutated molecules expressed by the majority of melanoma in vivo and most melanoma cell lines. In addition, unique minimal epitopic sequences play an immunodominant role in the context of specific HLA class I alleles. Since melanoma lesions from different patients often share expression of the same TA, and a minimal peptide sequence from a TA can cause immunologic changes in multiple patients, interest has grown in the development of TA-specific vaccines suitable for broad patient populations. Repeated in vitro stimulation of peripheral blood mononuclear cells (PBMC) with TA-derived epitopes can induce a high frequency of TA-reactive T-cells in melanoma patients. The same epitopes can also enhance TA-specific T-cell reactivity in vivo when administered subcutaneously in combination with Incomplete Freund's Adjuvant (IFA). Epitope based vaccinations, however, have not shown strong clinical efficacy unless combined with IL-2 administration. Attempts to increase the efficacy of these vaccines have combined specialised antigen-presenting cells or the administration of whole TA through DNA- or RNA-based vaccines with the intention of increasing antigen presentation and processing. Save for scattered reports, however, the success of these approaches has been limited and T-cell-directed vaccination against cancer remains at a paradoxical standstill whereby anticancer immunisation can be induced but it is not sufficient, in most cases, to induce tumour regression. Using melanoma as the standard model for immunotherapy, we will review various methods of T-cell-directed vaccination, the monitoring and analysis of the resulting immune response, and several clinical trials in which cancer vaccines have successfully induced immunisation. PMID- 11727536 TI - Use of bacteria as anticancer agents. AB - Historically, bacteria were used as oncolytic agents for malignant brain tumours. Advances in bacteriology and molecular biology have widened the scope of bacterial approaches to cancer therapy and various possibilities include the use of bacteria as sensitising agents for chemotherapy, as delivery agents for anticancer drugs, and as vectors for gene therapy. Bacterial toxins can be used for tumour destruction and cancer vaccines can be based on immunotoxins of bacterial origin. The most promising approaches are the use of genetically modified bacteria for selective destruction of tumours, and bacterial gene directed enzyme prodrug therapy. Knowledge gained from study of bacterial genomes forms an important basis of use of bacteria as anticancer agents. TAPET (Tumour Amplified Protein Expression Therapy) uses a genetically altered strain of Salmonella as a bacterial vector, or vehicle, for preferentially delivering anticancer drugs to solid tumours. Verotoxin 1 (VT1) of Escherichia coli has been used for ex vivo purging of human bone marrow of cancer cells before autologous bone marrow transplant. E. coli genes and enzymes have become part of well-known prodrug approaches to cancer in which inert prodrugs can be converted in vivo to highly active species. IL-4 fused with Pseudomonas exotoxin has been administered directly into malignant brain tumours and binds with high affinity to IL-4 receptors, which do not exist on normal brain cells, thus destroying a major part of the tumour without harming the normal brain tissue. It is in Phase I/II clinical trials in patients with glioblastoma. No ideal anticancer agent of bacterial origin that is applicable to all types of cancers has been discovered yet. The most promising approach to malignant brain tumours appears to be the use of genetically engineered bacteria that destroy the tumour selectively while sparing the normal brain tissue. PMID- 11727537 TI - Interleukin-1 receptor antagonist as therapy for inflammatory disorders. AB - IL-1 is a pivotal mediator of the immune response and has been implicated in inflammatory and infectious diseases. As a consequence, the administration of IL 1 receptor antagonist (IL-1ra), a recombinantly synthesised endogenous inhibitor of IL-1, has appeal as a therapeutic strategy in these conditions. To date, the largest clinical experiences with IL-1ra have been in the setting of sepsis and rheumatoid arthritis (RA). Like other anti-inflammatory agents that target a specific mediator, IL-1ra was found to lack efficacy when given in conjunction with standard therapy in patients with sepsis and septic shock. In contrast, recent studies enrolling patients with RA suggest that IL-1ra significantly ameliorates disease activity and retards joint destruction. Whether the respective lack of efficacy and success of IL-1ra in these two diseases is a result of differences in the pathologic processes involved, or reflects the nature in which the clinical studies were conducted, is unclear. Further, the effectiveness of IL-1ra compared to other anticytokine and conventional treatments in RA remains to be clarified. Nonetheless, the recent finding that IL 1ra has the ability to favourably influence a chronic inflammatory disease supports the hypothesis that inhibition of a single mediator of the immune response may have clinical impact. PMID- 11727539 TI - Oligonucleotide technologies: synthesis, production, regulations and applications. 29-30th November 2000, Hamburg, Germany. AB - The two-day meeting 'Oligonucleotide technologies', organised by IBC Global Conferences Limited and chaired by M Gait (Medical Research Council, UK), was the first event of this type in Europe. It covered a wide range of oligonucleotide based approaches useful for the identification and validation of new therapeutic targets as well as for the creation of novel oligonucleotide-based drugs. Recent progress in the preclinical and clinical development of new oligonucleotide-based therapeutics as well as new platforms and systems for DNA microarrays, allowing massively parallel gene expression studies, were discussed. Furthermore, new approaches for overcoming challenges in the large scale synthesis and functionalisation of oligonucleotides were revealed. The following meeting highlights are focussed on contributions devoted to the development of novel oligonucleotide-based drugs, such as antisense oligonucleotides, immunostimulatory CpG oligonucleotides and spiegelmers. Drug candidates discussed here are related to various indications, including cancer, infectious diseases and asthma. Besides highlighting of specific clinical candidates, emphasis is put on the basic principles of oligonucleotide-based drugs and approaches towards the improvement of their properties. PMID- 11727538 TI - G17DT--a new weapon in the therapeutic armoury for gastrointestinal malignancy. AB - G17DT or Gastrimmune, as it was formally known, is an antigastrin 17 immunogen producing neutralising high affinity antibodies directed against gastrin-17 (G17). Preclinical studies, initiated to identify biological functionality of G17DT-induced antibodies, confirmed that the antibodies both reduced G17 stimulated gastric acid secretion and inhibited gastrin from interacting with the CCK-2 receptor. Therapeutic efficacy of both passive and active immunisation with G17DT has been established in a number of tumour systems including both primary and metastatic disease. Furthermore, additive effects with 5-fluorouracil (5 FU)/leucovorin have been confirmed in both colon and gastric tumour models. Phase I/II studies in advanced gastrointestinal (GI) malignancies have shown no systemic or autoimmune reactions to active immunisation with G17DT. Use of an optimised dose has yielded a high proportion of responders (> 80%), with minimal side effects and antibody titres measurable within 2-4 weeks. Taken together these results suggest that the G17DT immunogen is a promising agent for the treatment of GI cancer and Phase III trials, currently underway, will definitively evaluate this early promise. PMID- 11727540 TI - Islet cell transplantation for the treatment of diabetes mellitus. AB - Diabetes mellitus is estimated to affect at least 16 million individuals in the United States and 135 million persons worldwide. It is a significant cause of morbidity and early mortality. The related expenses are astronomical with at least 15% of healthcare expenditures in the United States being used for the treatment of diabetes and its complications, a figure that approaches US$100 billion annually. The Diabetes Control and Complications Trial (DCCT) convincingly showed that intensive glucose management delays the onset and slows the progression of diabetic complications. Numerous studies have shown that pancreas transplantation not only delays the onset and progression of diabetic complications, but in some cases reverses some of the effects of diabetes. Human islet cell transplantation provides an alternative, less invasive alternative to whole organ transplantation. Human islet allotransplantation would only exacerbate the organ shortage, as recipients usually require islets from more than one pancreas. Xenotransplantation of porcine islets is a more attractive option; however, the recipient's immune response to xenografted tissue would be a formidable obstacle. Microencapsulation of the islets is a method of immunoisolation that would prevent the need for immunosuppressive drugs and the risks associated with their long-term use and have the potential to make xenoislet transplantation a clinical reality. PMID- 11727541 TI - O-raffinose-polymerised haemoglobin. A biochemical and pharmacological profile of an oxygen carrier. AB - Haemoglobin-based oxygen carriers (HBOCs) represent an interesting class of blood substitutes which are undergoing advanced clinical trials. The therapeutic goal of these compounds is to avoid or reduce blood transfusion in different surgical and medical situations of acute haemoglobin deficiency. Their main advantages include availability in large volumes, storage for prolonged periods, rapid administration (without typing and cross matching) and sterilisation by pasteurisation. Their main known disadvantages are reduced circulation half-life, haemodynamic and gastrointestinal perturbations, probably related to nitric oxide (NO) scavenging, free radical induction, and alterations of biochemical and haematological parameters (increases in liver enzymes levels, platelet aggregation). Cell-free o-raffinose cross-linked and oligomerised human haemoglobin (O-r-poly-Hb) (Hemolink, Hemosol, Canada) is a modified haemoglobin with molecular weight ranging from 32- > 500 kDa. Its affinity for oxygen appears lower than normal blood and an n (Hill coefficient) value of about 1 indicates a very low degree of co-operativity. Probably related to the low O2 affinity value and to the high molecular weight, O-r-poly-Hb has been shown to induce lesser haemodynamic perturbations than other first generation modified haemoglobins. This HBOC is in Phase III clinical trials in cardiac and orthopaedic surgery for perioperative haemodilution, at doses from 25 g (250 ml)-100 g (1000 ml). PMID- 11727542 TI - 15th Annual Scientific Meeting of the Society for Biological Therapy. 26-29 October 2000, Seattle, Washington, USA. AB - The 15th Annual Scientific Meeting of the Society for Biological Therapy (SBT) was held at the Four Seasons Olympic Hotel in Seattle, USA. The meeting was organised on behalf of the society by John A Thompson from the University of Washington (Seattle, USA), Michael B Widmer of Immunex Corp. (Seattle, USA) and Bernard A Fox from the Earle A Chiles Research Institute (Portland, Oregon, USA). The purpose of the organisation, which was founded in 1984 and currently has 300 members, is to bring together those diverse individuals actively investigating biologicals and biological response modifiers in the diagnosis and treatment of cancer, including clinicians and basic scientists from industry, government and academia. PMID- 11727543 TI - The 14th European Immunology Meeting--EFIS 2000. 23-27 September 2000, Poznan, Poland. AB - The 14th European Immunology Meeting--EFIS 2000, held in Poznan, Poland on 23-27 September 2000, was the last major meeting of European immunologists in the second millennium. This conference was intended to summarise past achievements and to present future prospects in immunology. The philosophy of the scientific program was to fuse fundamental and clinical immunology and give a chance for basic scientists and clinicians to discuss mutual topics in a general view. There were eight state-of-art lectures, 12 'meet an expert' sessions, 20 plenary sessions and 46 workshops. More than 900 works were presented. Significant interest was focused on several aspects of cancer immunology and immunotherapy. EFIS 2000 was accompanied by six pre-congress satellite symposia held in various Polish cities. The topics were, 'Heat shock proteins: immune, stress response and apoptosis' (Gdansk), 'Infectious immunity and vaccines' (Kazimierz Dolny), 'Mononuclear phagocytes in basic and clinical immunology' (Cracow), 'Immunology of reproduction' (Poznan), 'Primary immunodeficiencies' (Warsaw) and 'Glycoimmunology' (Wroclaw). PMID- 11727544 TI - Lentivirus and foamy virus vectors: novel gene therapy tools. AB - The aim of gene therapy is to modify the genetic material of living cells to achieve therapeutic benefit. Gene therapy involves the insertion of a functional gene into a cell, to replace an absent or defective gene, or to fight an infectious agent or a tumour. At present, a wide variety of somatic tissues are being explored for the introduction of foreign genes with a view towards treatment. A prime requirement for successful gene therapy is the sustained expression of the therapeutic gene without any adverse effect on the recipient. A highly desirable vector would be generated at high titres, integrate into target cells (including non-dividing cells) and have little or no associated immune reactions. Lentiviruses have the ability to infect dividing and non-dividing cells and, therefore, constitute ideal candidates for development of vectors for gene therapy. This review presents a description of available lentiviral vectors, including vector design, applications to disease treatment and safety considerations. In addition, general aspects of the biology of lentiviruses with relevance to vector development will be discussed. Recent investigations have revealed that foamy viruses, another group of retroviruses, are also capable of infecting non-dividing cells. Thus, foamy virus vectors are actively being developed in parallel to lentivirus vectors. This review will also include various aspects of the biology of foamy viruses with relevance to vector development. PMID- 11727545 TI - Novel approaches to allogeneic stem cell therapy. AB - Traditionally, allogeneic haematopoietic stem cell transplantation (SCT) has involved administration of myeloablative doses of chemotherapy and/or radiation that may cure many patients with haematologic diseases. The high morbidity and mortality associated with the intensive conditioning regimen limits allogeneic SCT to younger and healthier patients. However, it is now known that successful allogeneic SCT is dependent, at least in part, on the antitumour properties of the donor graft independent of the conditioning regimen. This potent 'graft versus tumour' (GVT) effect can now be exploited for clinical benefit. The best evidence of a direct GVT reaction comes from the use of donor leukocyte infusions (DLI). For many patients with relapsed leukaemia after allogeneic SCT, DLI re establishes complete and durable remissions. This has suggested a novel approach to allogeneic cell therapy (ACT) using non-myeloablative, but immunosuppressive conditioning regimens to permit engraftment of allogeneic stem cells and lymphocytes. Engrafted donor cells would then provide GVT activity in the setting of reduced conditioning regimen toxicity. The ability to minimise toxicity and maximise the immunologic GVT effect will make allogeneic transplantation applicable to patients typically ineligible for conventional allogeneic SCT. Response rates with this strategy have been impressive, although toxicity related to graft versus host disease (GVHD) and other complications remains a concern. Current trials have involved heterogeneous groups of patients using various conditioning regimens. Many issues remain unsettled, including identification of the most appropriate tumour targets and definition of the most effective, least toxic conditioning regimen. In addition, the durability of response is unknown. Nevertheless, the use of non-myeloablative conditioning and ACT may provide a new paradigm for allogeneic cell transplantation and the immunotherapy of cancer. PMID- 11727546 TI - Therapeutic application of chimeric RNA/DNA oligonucleotide based gene therapy. AB - Chimeric RNA/DNA oligonucleotides, or chimera, have emerged as a breakthrough technology for treating genetic disorders. Chimera have been shown to induce correction of point mutations in several genetic disease models without utilising the viral vectors. Recent studies of chimera-based gene therapy in genetic disease models are reviewed. Chimera were delivered intravenously, intramuscularly, intradermally, or topically with or without vehicles. Correction of the mutation at genotypic and phenotypic levels was assessed using various methods. The gene correction frequency varied, ranging from 1-40%. The resulting phenotype changes lasted longer than one year in some studies. The most dramatic phenotypic change is the reduction of serum bilirubin level by 50% in the Gunn rat, a model for Crigler-Najjar syndrome. Chimera based gene therapy has the potential to develop into powerful therapeutic modality for genetic diseases. PMID- 11727547 TI - Prodrugs in nasal drug delivery. AB - Prodrugs have been used to overcome poor solubility, insufficient stability, incomplete absorption across biological membranes and premature metabolism to active species. This review examines the importance of various physicochemical factors affecting nasal absorption of drugs. Novel trends in nasal prodrug development in the areas of targeted delivery to the CNS and selective targeting of the nutrient transporter system of the nasal mucosa have received considerable attention. PMID- 11727548 TI - Heat shock proteins: novel therapeutic tools for HIV-infection? AB - Heat shock proteins (Hsps), cyclophilins (Cyps) and FK binding proteins (FKBPs) form a family of intracellular chaperone molecules that facilitate protein folding and assembly. These stress proteins are selectively expressed in cells in response to a range of stimuli, including heat, lymphokine and microbial/viral infections. This review discusses the role of stress proteins in the HIV-1 viral life cycle, with regard to the development of specific Hsp-based therapeutic strategies against HIV-1 infection. Cumulative findings are cited implicating CypA, Hsp27, Hsp70 and FKBPs in host cell and viral activation, viral entry, assembly or formation of infectious virions. Biological response modifiers that show specific high-affinity interactions with Cyp, FKBPs and Hsps, including cyclosporins, FK-506 and cyclopentenone prostaglandins respectively, may block HIV-1 replication and infection, providing novel HIV-1 therapeutic strategies. Moreover, Hsp binding to viral complexes can enhance antiviral immunity, including natural killer (NK), antibody-dependent (ADCC), gamma delta T-cell and cytotoxic T-lymphocyte (CTL) activities against HIV-1 infected cells. The ability of Hsps to interact with HIV-1 viral proteins, combined with their inherent adjuvant and immunogenic properties indicates that Hsps may also serve as vehicles for antigen delivery and the design of AIDS vaccines. PMID- 11727549 TI - Restenosis and gene therapy. AB - Atherosclerosis is one of the main causes of mortality and morbidity in westernised countries. Treatment of symptomatic atherosclerosis by angioplasty involves major vascular responses such as neointima formation and constrictive vascular remodelling leading to restenosis. Stent placement prevents vasoconstriction but is associated with in-stent neointima formation. Therefore, stent placement requires adjunctive therapy. In this review we discuss the potential of local gene therapy for restenosis. More particularly, we focus on strategies to inhibit smooth muscle cell (SMC) proliferation and migration, prevent thrombosis, decrease oxidative stress in the arterial wall and enhance re endothelialisation associated with adaptive remodelling. The potential of different vector systems and devices for local gene transfer in the arterial wall is discussed. PMID- 11727550 TI - Thymic hormones in cancer diagnostics and treatment. AB - The thymus is an endocrine organ. A unified, physiological concept of humoral regulation of the immune response emerged in the last three decades. The thymus is the primary major site of production of immunocompetent T-lymphocytes from their haematopoietic stem cells. The thymus provides a superior humoral microenvironment for the development of immunocompetent T-lymphocytes. Although yolk sac derived pre-T stem cells enter the thymus using a homing receptor, the immigration process requires also secretion of a peptide, called thymotaxin by the cells of the reticulo-epithelial (RE) network. This complex process requires direct cell to cell, receptor based interactions, as well as in situ paracrine information via the numerous cytokines and thymic hormones produced by the RE cells of thymic microenvironment. Thymic hormones induce in situ T-lymphocyte marker differentiation, expression and functions. These polypeptide hormones have also been shown by means of immunocytochemistry to localise in the RE cells of the thymic cellular microenvironment. Based on the complexity of the intrathymic maturation sequence of T-lymphocytes and the increasing numbers of T-lymphocyte subpopulations that are being identified, it would be surprising if a single thymic humoral factor could control all of the molecular steps and cell populations involved. Rather, it would appear that the control of intrathymic T lymphocyte maturation and functional maturation involves a complex number of thymic-specific factors and other molecules that rigidly control the intermediary steps in the differentiation process. Thymosin fraction 5 (TF5) and its component polypeptides influence a variety of lymphocyte properties including cyclic nucleotide levels, migration inhibitory factor production, T-dependent antibody production and expression of certain surface maturation/differentiation markers. Recently, thymic hormones, mostly thymosins have been employed not only in neoplasms' early detection but also in clinical trials to strengthen the effects of immunomodulators in immunodeficiencies, autoimmune diseases and neoplastic malignancies. Combined chemoimmunotherapeutical antineoplastic treatment seems to be useful. Generally, haematopoietic toxicity of every chemotherapeutical clinical trial can be reduced significantly by the immunotherapy, compared to 50% in patients treated with chemotherapy alone. PMID- 11727551 TI - The antimetastatic and immunomodulating activities of ginseng minor glycosides. PMID- 11727552 TI - Caspase-dependent cytotoxicity of anti-DNA autoantibodies. PMID- 11727553 TI - On the noncyclic mechanism of cholinesterase-mediated catalysis. PMID- 11727554 TI - Modification of the heme electronic structure induced by complex formation with nitric oxide and the atomic backbone dynamics at room temperature. PMID- 11727555 TI - The polyprotein gag of retroelement gypsy can form multimeric complexes when expressed in bacterial system. PMID- 11727556 TI - Stable mRNA-ribosome-polypeptide complexes and their use in functional screening of protein libraries. PMID- 11727557 TI - A general approach to renaturation of recombinant proteins produced as inclusion bodies. PMID- 11727558 TI - The involvement of actin in the formation of root pressure. PMID- 11727559 TI - Participation of free radicals in photoreduction of pterins and folic acid. PMID- 11727560 TI - Supramolecular structure of nanodiamond particles and obelin built up on a two dimensional plate. PMID- 11727561 TI - Circulating immune complexes in blood of patients with positive family history of schizophrenia. PMID- 11727563 TI - The region of rat chromosome 10 (the ngfr gene locus) is associated with blood pressure increase in response to emotional stress. PMID- 11727562 TI - Antiherpetic activity of dimeric derivatives of netropsin. PMID- 11727565 TI - Structural changes in DNA molecules induced by ultralow-intensity field of thermalized neutrons. PMID- 11727564 TI - The conserved characteristics of DNA-binding domains belonging to the homeodomain class that are associated with coadaptive substitutions of amino acid residues. PMID- 11727566 TI - Frequency-dependent effect of insulin on myocardial contractility in active ground squirrel Citellus undulatus in different seasons. PMID- 11727567 TI - Nonstationary dynamics of bacterial population waves. PMID- 11727568 TI - Both sides now. Many paths bring people to research. PMID- 11727569 TI - Change and rumors of change. PMID- 11727570 TI - On approval. PMID- 11727571 TI - Tenacious tenofovir struts its stuff in a virtual ICAAC. PMID- 11727572 TI - Direct to consumer advertising; the right tool in the wrong hands? PMID- 11727573 TI - Reducing abortions is a public health issue. PMID- 11727574 TI - Microbiological aspects of peritonitis in patients undergoing chronic peritoneal dialysis at the dialysis unit of Sri Jayawardenapura General Hospital. AB - BACKGROUND: Peritoneal dialysis (PD) is an established form of therapy in the management of end stage renal disease. Peritonitis is the main complication of PD. OBJECTIVES: To study the incidence and microbial aetiology of peritonitis in patients undergoing chronic PD at the dialysis unit of Sri Jayewardenapura General Hospital (SJGH); to assess the diagnostic value of the Gram's stain; and to study the relationship of the total white cell count of effluent to peritonitis. DESIGN: A prospective study over three months. SETTING: Dialysis unit of SJGH. PATIENT POPULATION: The study involved 18 patients undergoing manual intermittent peritoneal dialysis (IPD), 4 patients undergoing chronic ambulatory peritoneal dialysis (CAPD), and 1 patient undergoing nocturnal intermittent peritoneal dialysis (NIPD). MEASUREMENTS: Clinical presentation of patients with peritonitis; total and differential white blood cell counts of effluent samples; Gram stain and culture of the centrifuged deposit to determine microbial aetiology; incidence of peritonitis in different categories of dialysis. RESULTS: 32 samples were examined from patients on IPD, and 17 from patients on CAPD. In IPD most episodes were due to Gram negative organisms whereas in CAPD most episodes were due to Gram positive organisms. Sensitivity of Gram's stain in relation to culture was 32.4%. 98% of effluent samples had white blood cell counts of > 100/ml and none showed neutrophil counts of < 49%. CONCLUSIONS: The incidence of IPD associated peritonitis was 11.1 episodes per patient year, and the incidence of CAPD associated peritonitis was 14 episodes per patient year. Flavobacterium spp. were the predominant organisms in IPD associated peritonitis, whereas CAPD associated peritonitis was commonly caused by coagulase negative staphylococci. Gram's stain was not useful in the initial identification of the causative agent, but the white cell and neutrophil counts were found to be sensitive indicators of peritonitis. PMID- 11727575 TI - Neurological complications of measles. AB - OBJECTIVE: To study the clinical features and course of neurological complications of measles. DESIGN: Prospective study. SETTING: Neurology Unit, Teaching Hospital, Kandy. PATIENTS: 10 patients admitted with neurological complications of measles during an outbreak. MEASUREMENTS: Assessment of clinical features and the neurological investigations. RESULTS: 6 had CSF IgM antibodies confirming a recent measles infection. 7 patients had encephalitis and 3 had myelitis. Complications had appeared 5 to 14 days after appearance of the rash. All patients with encephalitis had depressed level of consciousness (Glasgow coma scale 2 to 11) with seizures, 3 patients had lateralising signs, and assisted ventilation was required in 4. Cranial CTs were either normal or showed mild cerebral oedema. CSF analysis showed either normal or mildly raised protein without a cellular reaction. EEG in all 7 revealed diffuse delta activity. During the follow up period of 1 to 12 weeks, all showed a gradual improvement, except one who succumbed to respiratory complications of assisted ventilation. All others were left with some residual disability when last seen. Three patients with myelitis had symmetrical paraparesis. CSF protein was 60 to 80 mg/ml without a cellular reaction. All three improved without residual disability. CONCLUSIONS: Measles encephalitis is a life threatening complication. All three myelitis patients recovered completely within 5 to 8 weeks. PMID- 11727576 TI - Children of imprisoned mothers. AB - OBJECTIVES: To describe the problems faced by children during separation from their imprisoned mothers, and evaluate the health of children who accompanied their mothers into prison. DESIGN: A prospective observational study. SETTING: Welikada Prison, Colombo, Sri Lanka. METHODS: 200 randomly selected mothers who had left their children at home were interviewed using a questionnaire. During a period of 8 months 30 children living with their mothers in prison underwent physical and developmental examination and tuberculin testing. The living conditions within the prison were evaluated. RESULTS: During 18 months from January 1999, 4089 women were imprisoned. 88% were remanded, 20% awaited trial for more than one year in prison. 2416 were mothers. 1411 had at least one child under 12 years of age. The 200 mothers interviewed had 262 children under 12 years at home. Their care arrangements were: a relative (69%), father (16%), older sibling (4%), religious organisation (2.7%), neighbour (1.3%). None had received social services support. 70 children accompanied mothers into prison. In the 30 children followed up regularly 23% had scabies, 10% pediculosis, and 7% impetigo. No severe malnutrition was found and screening for tuberculosis was negative. 70% were breastfed. The child-friendly dormitory was inadequate to accommodate all children. CONCLUSION: Care arrangements and schooling were affected and no counselling services were provided during the imprisoned mothers' absence. The children within the prison enjoyed close bonds with the mothers and their physical needs were met. The child's best interest had not always been considered by court when deciding on custody during the mothers' imprisonment. PMID- 11727577 TI - Ethical regulation and the new reproductive technologies in Sri Lanka: perspectives of ethics committee members. AB - OBJECTIVE: To report on part of a pilot study done to explore ethical responses to the new reproductive technologies in Sri Lanka. DESIGN: Semi-structured interviews carried out with the members of three committees responsible for ethical review of medical research. Members were asked for their views on the ethical, social and legal implications of the new reproductive and genetic technologies. Members' responses were subject to a simple content analysis. CONCLUSIONS: There was broad acceptance of the new technologies among respondents, but anxieties about potential abuses. Respondents felt that a national committee should regulate practice and monitor future policy. PMID- 11727578 TI - Overcrowding and "floor" patients in state hospitals: institutions can make a difference. PMID- 11727579 TI - Examination of a scene of crime; do we need to amend section 370 of the Code of Criminal Procedure Act of Sri Lanka? PMID- 11727580 TI - The need for an Institute of Hospital Management. AB - The monitoring mechanism of activities of the government preventive health sector is better than that of the curative sector, although much more money is spent on the latter. Recent initiatives for monitoring the curative sector should be strengthened by establishment of an institute of hospital management. The need for such an institute has been recognised for about 15 years. This need has been further emphasised by recent studies and discussions. PMID- 11727581 TI - Body stalk anomaly. PMID- 11727582 TI - Complete colonic duplication in an infant. PMID- 11727583 TI - Causes of headache in a Sri Lankan general practice. PMID- 11727584 TI - Periodic changes in behaviour in a child with pseudohypoparathyroidism. PMID- 11727585 TI - Fulminant heptic failure in a child following paracetamol overdosing. PMID- 11727586 TI - A prospective hospital-based study of dengue haemorrhagic fever. PMID- 11727587 TI - Thromboprophylaxis as routine for Sri Lankan surgical patients: where is the evidence? PMID- 11727588 TI - Yet another form of child abuse? PMID- 11727589 TI - Collection and transport of urine for culture. PMID- 11727590 TI - A review and analysis of the clinical- and cost-effectiveness studies of comprehensive health promotion and disease management programs at the worksite: 1998-2000 update. AB - PURPOSE: This article is the fifth in a series of critical reviews of the clinical effectiveness and cost-effectiveness studies of comprehensive, multifactorial health promotion and disease management programs conducted in worksites. As with the previous reviews, the purpose of this article is to review and assess the randomized control trials that have focused on both clinical and cost outcomes of worksite health promotion and disease management programs. For this current review, a new category of quasi-experimental studies has been added because this represents a major new trend in such interventions over the last 2 years. Comprehensive worksite programs are those that provide an ongoing, integrated program of health promotion and disease prevention that integrates specific components into a coherent, ongoing program that is consistent with corporate objectives and includes program evaluations of both clinical and cost outcomes. DATA SOURCES: A comprehensive search was conducted using a multistage process that included MEDLINE, ERIC, ADI, EDGAR, CARL, Inform, and Lexis-Nexis databases and direct inquiries to worksite researchers. INCLUSION AND EXCLUSION CRITERIA: The search identified 27 new studies to which the following inclusion criteria were applied: research conducted in the United States; results published in English; methodological quality of nonexperimental (pre- and postmeasures but no comparison group); quasi-experimental to randomized control trials; and both clinical and cost outcomes. Exclusion criteria were studies outside of the United States; non-English publications; and clinical or cost outcomes only. Fifteen studies remained for this review. FINDINGS EXTRACTION METHODS: Findings extraction and analysis of the 15 studies was done by extracting the relevant population, intervention design, clinical results, and cost outcomes from the published article. As in previous reviews, findings are summarized in a table format that extracts and describes each study by the following: (1) study author(s); (2) corporate site; (3) purpose of the evaluation; (4) employee population; (5) percentage of program participants; (6) number of employees included in the evaluation; (7) brief description of the intervention; (8) evaluation design; (9) evaluation period; (10) outcomes; (11) research rating; and (12) findings. FINDINGS SYNTHESIS: Based on these 15 studies, a methodological critique was conducted with brief reference to appropriate prior studies. Conclusions regarding study quality and new trends over the time period of 1998 to 2000 are discussed. MAJOR CONCLUSIONS: Results from randomized clinical trials and quasi-experimental designs suggest that providing individualized risk reduction for high risk employees within the context of comprehensive programming is the critical element of worksite interventions. Despite the many limitations of the current methodologies of the 15 new studies, the vast majority of the research to date indicates positive clinical and cost outcomes. PMID- 11727591 TI - Association of awareness, intrapersonal and interpersonal factors, and stage of dietary change with fruit and vegetable consumption: a national survey. AB - PURPOSE: To examine associations of awareness, intrapersonal and interpersonal factors, and stage of change with consumption of fruits and vegetables. DESIGN: Nationally representative, random digit dial survey conducted in 1997 with a response rate of 44.5%. Psychosocial correlates of fruit and vegetable consumption were assessed using regression analyses. SETTING: United States. SUBJECTS: A total of 2605 adults who were 18 years and older. MEASURES: Awareness of the "5 A Day for Better Health" program and its message, along with stage of change; taste preferences; self-efficacy; and perceived benefits, barriers, threats, social support, and norms related to fruit and vegetable consumption. RESULTS: Awareness and intrapersonal and interpersonal factors explained 24% of the variance in fruit and vegetable consumption beyond the 9% explained by demographic characteristics. Knowledge of the 5 A Day message was associated with a 22% increase in fruit and vegetable consumption. Self-efficacy for eating fruits and vegetables and taste preferences (affect) were the factors most consistently and strongly associated with both higher consumption and higher likelihood of being in action or maintenance stages of change. Affect and perceived barriers were more strongly associated with increased vegetables and salad than fruit. CONCLUSIONS: Dietary intervention programs to increase fruit and vegetable consumption should emphasize the 5 A Day message, increased self efficacy, and ways to make vegetables more palatable and easily accessible. Understanding the factors that influence dietary choices should be used when designing dietary interventions. PMID- 11727592 TI - Weight gain as a barrier to smoking cessation among military personnel. AB - PURPOSE: To assess the relationships between active-duty military status, military weight standards, concern about weight gain, and anticipated relapse after smoking cessation. DESIGN: Cross-sectional study. SETTING: Hospital-based tobacco cessation program. SUBJECTS: Two hundred fifty-two enrollees, of 253 eligible, to a tobacco cessation program in 1999 (135 men, 117 women; 43% on active duty in the military). MEASURES: Independent variables included gender, body mass index (weight/height2), and military status. Dependent variables included about weight gain with smoking cessation and anticipated relapse. RESULTS: In multivariate regression analyses that controlled for gender and body mass index, active-duty military status was associated with an elevated level of concern about weight gain (1.9-point increase on a 10-point scale; 95% confidence interval [CI], 1.0- to 2.8-point increase), as well as higher anticipated relapse (odds ratio [OR] = 3.6; 95% CI, 1.3 to 9.8). Among subjects who were close to or over the U.S. Air Force maximum allowable weight for height, the analogous OR for active-duty military status was 6.9 (p = .02). CONCLUSIONS: Occupational weight standards or expectations may pose additional barriers for individuals contemplating or attempting smoking cessation, as they do among active-duty military personnel. These barriers are likely to hinder efforts to decrease smoking prevalence in certain groups. PMID- 11727593 TI - Barriers among nurses to providing smoking cessation advice to hospitalized smokers. AB - Nine focus groups were conducted with 75 staff nurses in three hospitals to determine the low participation rate of nurses in a smoking cessation programs directed at patients. Nurses felt that hospitalization was an appropriate time to offer quit-smoking advice to patients, particularly those with smoking related diseases. However, many felt advice should be given only to those receptive to it. The most common barriers to providing smoking cessation advice were lack of concrete techniques, or referral to provide and fear of alienating patients who were not receptive to hearing advice. PMID- 11727594 TI - Measuring stage of change for assessing readiness to increase fruit and vegetable intake among 18- to 24-year-olds. AB - PURPOSE: To examine the appropriateness of the five-stage schema developed for addictive behaviors when applied to nonaddictive behaviors such as fruit and vegetable consumption. DESIGN: A cross-sectional mail survey was conducted. SETTING: Self-administered questionnaires were mailed to the home addresses of respondents recruited in 10 states and returned upon completion. SUBJECTS: In all, 116 male and 185 female young adults (response rate = 70%) completed the questionnaires. Most respondents were Caucasian (74.4%) and college students (62.2%). MEASURES: Statistical analyses included analysis of variance with Scheffe's test, Pearson correlation and Cronbach's alpha coefficients, factor analysis, and cluster analysis. RESULTS: Staging algorithms for fruits and vegetables were developed according to the five-stage schema, and their construct validity was verified by the incremental trend of food intakes. The algorithms identified more than 80% of the subjects as precontemplators, preparers, or maintainers for both food groups. This distribution pattern agreed well with the three-factor solution generated from the stages of change questionnaire, for which internal reliability and validity were confirmed. Seven cluster profiles of the respondents were derived based on the three generated factors and appeared to represent stage subgroupings of those factors. The two food groups shared marked similarities regarding the profiling of the clusters, which was validated by comparing food intake. The results, however, cannot be generalized because of limitations involved in the current sample. CONCLUSIONS: The original five stages of change and traditional staging measures may need to be adapted and/or extended for dietary change. PMID- 11727595 TI - Predicting physical activity promotion in health care settings. AB - PURPOSE: To test the ability of the theory of planned behavior (TPB) in predicting the stage of change for physical activity promotion by mental health professionals. DESIGN: Six-month prospective questionnaire study. SETTING: One mental health trust in the East Midlands, United Kingdom. SUBJECTS: Three hundred ninety-four mental health professionals (men, n = 131; women, n = 263) of an initial sample of 477 participated in the study (83% response rate). MEASURES: Attitudes, subjective norms, intentions, perceived behavioral control, and stage of change were measured at the first wave of data collection. Stage of change was also assessed 6 months later. Data were analyzed using structural equation modeling. RESULTS: Intention and stage of change were successfully predicted from TPB variables. Overall, 27% of the variance in self-reported stage of promoting physical activity was explained by the model. Sixty-one percent of the variance in intention to promote physical activity was explained. When included, past behavior was the strongest predictor of both intention and stage of change and attenuated all other path coefficients. Past behavior improved the predicted variance in intention by 11% and stage by 6%. CONCLUSIONS: The TPB variables of attitude, subjective norms, perceived behavioral control, and intention predict stage of change of physical activity promotion in a health care setting. However, promoting physical activity in the past had a sizable effect on predicting subsequent promotion. Due to unequal distribution across stages, the stage model's application to understanding the behavior of health professionals may be limited. PMID- 11727596 TI - Fiscal patience. Word still out on innovis' cost savings in N.D. PMID- 11727597 TI - Eye on info. Clinical-care IT still the final frontier. PMID- 11727598 TI - Eye on info. Vendors say they're ready to deliver. PMID- 11727599 TI - Eye on info. Trying to go beyond order entry. PMID- 11727600 TI - Eye on info. Bringing order to doctors' orders. PMID- 11727601 TI - Relief on a shoestring. Amid fiscal pressure, feds look at Medicare deregulation as way to help providers. PMID- 11727602 TI - Eye on info. Cutting clinical risk and expense. PMID- 11727603 TI - Risk-management awards. Focus on stroke care. Hospitals' programs deliver clinical, financial benefits. PMID- 11727604 TI - Risk-management awards. Prescribing quality. Docs use technology, office redesign to make gains. PMID- 11727606 TI - Medication errors related to potentially dangerous abbreviations. PMID- 11727605 TI - 50 years of progress in health care quality and safety. PMID- 11727607 TI - Changes in proficiency test monitoring and follow up for laboratories. PMID- 11727608 TI - Preventing suicide: how nurses can help. PMID- 11727609 TI - Completing competency assessments in home care. PMID- 11727610 TI - ['Benign' meningioma]. AB - Meningioma was diagnosed in four women, aged 40, 24, 41 and almost 75 years, respectively. The first of these patients was treated with surgery, the second and third patients underwent surgery followed by conventional radiotherapy because of a tumour residue or dural tail, and the last patient was treated with stereotactic radiosurgery. They recovered well and were followed by means of regular outpatient check-ups. Twenty percent of all primary brain tumours are meningiomas, over 90% of which are benign. Nevertheless, a large hospital-based population study showed a 5-year survival rate of only 70%. Microsurgery is usually the treatment of first choice. However, in about 25% of cases, excision is incomplete and tumour growth almost always continues. Further surgery influences prognosis unfavourably. New sophisticated radiation techniques help to control tumour progression in about 80-90% of cases. This success, however, may be associated with new cranial nerve deficits or panhypopituitarism. Prospective, comparative studies are not available. PMID- 11727611 TI - [Preoperative examination: anamnesis and physical examination mandatory]. AB - Preoperative assessment has been subject to discussion for many years. What should be done and by whom? Fortunately there is a general consensus that routine laboratory and function tests are not only of no benefit to the patient but are also wasteful of resources. The Preoperative Assessment Commission of the Dutch Ministry of Health, Welfare and Sport, supports the initiative for preoperative assessment clinics run by anaesthesiologists. Yet in their advice, this Commission has proposed screening by means of an abridged questionnaire. Efforts to validate this questionnaire have demonstrated that in practice it was not helpful. Therefore it seems justified to conclude that preoperative screening should not fundamentally differ from that performed during a basic clinical examination. PMID- 11727613 TI - [Meningiomas: prognostic relevance of histopathologic and genetic markers]. AB - The majority of meningiomas are histologically benign tumours. Location and invasion of tumour tissue in adjacent structures may hamper radical resections and give rise to recurrences. The rise in human life expectancy has prolonged the postoperative period and thus the risk of tumour recurrence has increased markedly. Infiltration in brain tissue and mitotic activity are important histologic features which negatively influence the disease-free duration of the postoperative period. Molecular studies of relevant genetic defects involved in meningioma are currently underway, but as yet these are of little clinical relevance. PMID- 11727612 TI - [Letters of condolence to the relatives of patients]. AB - Most people who die in the Netherlands receive medical care during the final stage of their life. Since the 19th century, when a sombre prognosis was often the only medical intervention available, sophisticated diagnostic procedures and curative victories have forced elementary bedside manners into retreat. However, modern medicine is rediscovering its boundaries; the inevitability of death and the human tragedy of dying. The renewed interest in letters of condolence from doctors to the family of their deceased patients is an example of this. These letters provide the physician with an opportunity for worthy and effective behaviour under trying circumstances. PMID- 11727614 TI - [Intraocular retinoblastoma: new therapeutic options]. AB - Retinoblastoma is the most frequently occurring primary intraocular malignant tumour in children (12-15 new patients per year in the Netherlands). It occurs in one or two eyes. Bilateral retinoblastoma, which occurs in 40% of the cases, is always hereditary; unilateral retinoblastoma, which is found in 60% of cases, is hereditary in 10% of these cases. The presenting symptoms are: leucocoria, strabismus or a red, painful eye. Early detection of retinoblastoma is important for the chance of survival, the visual prognosis and preservation of the eye. The choice of treatment is based on the risk of metastases, the diameter and the location of the tumour, the age of the patient, the heredity and the visual prognosis. Nowadays, treatment more often consists of a combination of techniques. Enucleation is carried out when a large tumour fills over half of the globe; often this is the only possible treatment. Small tumours (diameter and thickness < 2 mm) in the centre of the retina can be treated with laser therapy and those in the peripheral retina by cryotherapy. Small to medium-sized tumours (< 8 mm diameter) can be treated with thermochemotherapy: systemic chemotherapy and laser hyperthermia, if necessary with adjuvant laser therapy or brachytherapy. Medium-sized tumours (< 8 mm thick) can be treated with just brachytherapy, sometimes preceded by chemoreduction. PMID- 11727615 TI - [Roaming through methodology. XXXV. Bias in prevalent cohort studies of the natural course of disease]. AB - The natural history of infectious diseases with a long asymptomatic incubation period has mainly been studied in cohorts of individuals already infected at study entry: the so-called prevalent cohort study. Because the time of infection is usually unknown in the prevalent cohort, in standard survival analysis it is common to use the time since entry into the cohort instead of the time since infection to study risk factors for disease progression. However, the use of the time since study entry may bias results. The two most important sources of bias are onset confounding and differential length-bias sampling. Because bias may occur, results derived from a prevalent cohort are not directly comparable to results derived from an incident cohort where the moment of infection is known. PMID- 11727616 TI - [Diagnostic image (63). Paraganglioma]. AB - A 33-year-old woman was examined for a non-tender mass in the neck. Imaging findings were consistent with paraganglioma. The diagnosis was confirmed at surgery. PMID- 11727617 TI - [Short preoperative health assessment proposed by the Health Council not useful in clinical practice]. AB - OBJECTIVE: To evaluate the value of the short questionnaire drawn up by the Dutch Health Council for the classification of patients to an ASA class (a 5-point scale according to the American Society of Anesthesiologists, on which the patient's preoperative physical condition can be scored) and to propose an anaesthesia care plan for surgery patients aged between 16 to 40 years old. DESIGN: Observational. METHODS: From June 1999 through to May 2000, all 2090 preoperative patients aged 16 to 40 years at the academic hospital of Utrecht, the Netherlands, were asked to complete the Dutch Health Council's short questionnaire. In addition, the usual extensive preoperative health assessment was carried out on these patients. Of the 379 (18%) who were found by the short questionnaire to be 'healthy', 100 were selected. A panel of 10 anaesthesiologists was asked to preoperatively evaluate these 100 patients twice: once using the short questionnaire and once using the usual extensive health evaluation. The primary outcome was the percentage of patients who could be classified to an ASA class and for whom an anaesthesia care plan could be drawn up. The secondary outcome was the information judged by the anaesthesiologists to be either 'redundant' or 'missing' when drawing up an anaesthesia care plan. RESULTS: Using the short questionnaire, 63% of the patients could not be assigned to an ASA class, compared to 22% with the extensive health evaluation (p < 0.0001). On the basis of the information obtained with the short questionnaire, it was not possible to draw up an anaesthesia care plan for any of the patients, while the extensive health evaluation enabled an anaesthesia care plan to be drawn up for 65% of the patients (95% CI: 62-68%). Using the missing information deemed 'necessary', recommendations were made for the minimum scope of a preoperative health assessment for use in patients aged 16 to 40 years old. CONCLUSION: The short questionnaire as proposed by the Dutch Health Council was not found to be useful in practice. PMID- 11727618 TI - [Two neonates with vesicular skin lesions due to incontinentia pigmenti]. AB - In two neonate girls with vesicular skin lesions, incontinentia pigmenti (Bloch Sulzberger syndrome) was diagnosed. This rare X-linked dominant ectodermal disease can cause abnormalities in several organ systems. Most prominent are the dermatological abnormalities, developing in 4 stages: the vesicular stage, the verrucous stage, the hyperpigmentation phase, and the atrophic phase. In addition to the cutaneous manifestations, many patients have anomalies of nails, hair and teeth. Serious related abnormalities of the eyes (intraocular vasculopathy) and central nervous system (convulsions, mental retardation) may occur. In 1989 the locus for the incontinentia pigmenti mutation was shown to be present on Xq28. Recently it was shown that the causative mutation is located on the NEMO ('NF kappa B essential modulator') gene. A NEMO knock-out mouse model shows a dermatopathy of a transient nature, resembling the skin lesions in patients with incontinentia pigmenti. PMID- 11727619 TI - [Injury to the right A. pulmonalis due to blunt thoracic trauma]. AB - Immediately after a moped accident a 16-year old patient was haemodynamically unstable. Following adequate drainage of a haematopneumothorax in the hospital the patient collapsed into a shock. Thoractomy revealed a tear of the right A. pulmonalis. The hilus was clamped and a lobectomy was performed. After this the patient was haemodynamically stable; due to bone and nerve damage he was transferred to a rehabilitation centre. Laceration of the A. pulmonalis as a consequence of blunt thoracic trauma is rare and mostly fatal. In the case of a patient with a massive haematothorax rapid transport to the nearest hospital is preferable to thorax drainage at the site of the accident. PMID- 11727620 TI - ['Caring always'; physicians' expression of condolences to the relatives of patients]. AB - According to the WHO definition, palliative care does not end with the death of the patient, but also includes supporting the family members in dealing with the loss. A recent article refers to the importance of physicians writing letters of condolences. The letter of condolences marks the end of the treatment relationship. Writing such a letter can assist the physician in distancing him/herself from the death of his/her patient, and the letter can also help the family adjust to life without the deceased patient. The letter can also remove any distrust that may exist towards the medical profession. Reasons for not writing a letter of condolences include an own feeling of failure, administrative difficulties, indifference or incapacity. Nevertheless, writing a letter of condolences is an integral part of the care provided by the physician to the patient and his/her relatives. PMID- 11727621 TI - [Subacute combined degeneration of the spinal cord due to vitamin B12 deficiency: easy diagnosis, effective therapy]. PMID- 11727622 TI - [Subacute combined degeneration of the spinal cord due to vitamin B12 deficiency: easy diagnosis, effective treatment]. PMID- 11727623 TI - [Analysis of expert reports from a general practice expert in disciplinary cases and malpractice claims against family practitioners (1994-2000): a lesson for clinical practice]. PMID- 11727624 TI - [Activated protein C, coagulation, inflammation and treatment of severe sepsis]. PMID- 11727625 TI - [Assessment of left ventricular function and volume by myocardial perfusion scintigraphy--comparison of two algorithms]. AB - AIM: Left ventricular volume and function can be computed from gated SPECT myocardial perfusion imaging using Emory Cardiac Toolbox (ECT) or Gated SPECT Quantification (GS-Quant). The aim of this study was to compare both programs with respect to their practical application, stability and precision on heart models as well as in clinical use. METHODS: The volumes of five cardiac models were calculated by ECT and GS-Quant. 48 patients (13 female, 35 male) underwent a one day stress-rest protocol and gated SPECT. From these 96 gated SPECT images, left ventricular ejection fraction (LVEF), end-diastolic volume (EDV) and end systolic volume (ESV) were estimated by ECT and GS-Quant. For 42 patients LVEF was also determined by echocardiography. RESULTS: For the cardiac models the computed volumes showed high correlation with the model-volumes as well as high correlation between ECT and GS-Quant (r > or = 0.99). Both programs underestimated the volume by approximately 20-30% independent of the ventricle size. Calculating LVEF, EDV and ESV, GS-Quant and ECT correlated well to each other and to the LVEF estimated by echocardiography (r > or = 0.86). LVEF values determined with ECT were about 10% higher than values determined with GS-Quant or echocardiography. The incorrect surfaces calculated by the automatic algorithm of GS-Quant for three examinations could not be corrected manually. 34 of the ECT studies were optimized by the operator. CONCLUSION: GS-Quant and ECT are two reliable programs in estimating LVEF. Both seem to underestimate the cardiac volume. In practical application GS-Quant was faster and easier to use. ECT allows the user to define the contour of the ventricle and thus is less susceptible to artifacts. PMID- 11727626 TI - [Prevalence of iodine- and thyroglobulin-negative findings in differentiated thyroid cancer.A retrospective analysis of patients treated from 1951 to 1998 in university hospital]. AB - AIM: The prevalence of iodine- and thyroglobulin-negative findings was evaluated in all patients with differentiated thyroid cancer (DTC) treated from 1961 until 1998 at the Department of Nuclear Medicine, University Hospital Hamburg Eppendorf. METHODS: A total of 490 patients with papillary thyroid cancer (PCA) and 242 patients with follicular thyroid cancer (FCA) were analyzed retrospectively. Patients were divided into four groups: 1: no recurrence, 2: recurrent disease, 3: primary metastatic/progressive disease and 4: inconclusive follow-up. Results of iodine scan, serum-TG, and additional imaging modalities as well as histology were compared in all patients. RESULTS: 21/490 (4.3%) of patients with PCA and 16/242 (6.6%) with FCA suffered from recurrent disease. 62/490 (12.7%) of patients with PCA and 59/242 (24.4%) with FCA had primary metastatic/progressive disease. 12/21 patients with PCA and 12/16 with FCA showing up with recurrent disease had a negative iodinescan. 11/21 of patients with PCA and 4/16 with FCA and tumor recurrence had negative serum-TG levels. 14/62 patients with PCA and 14/59 with FCA presenting with primary metastatic/progressive disease had negative iodinescan. 14/62 patients with PCA and 6/59 with FCA had negative serum-TG. CONCLUSION: The prevalence of iodine negative recurrent/metastatic disease is in accordance to the literature, whereas the prevalence of TG-negative recurrent/metastatic was noted higher than reported previously. Thus, the commonly used follow-up scheme of DTC is confirmed. However, iodine scan should be regularly performed in patients with high risk of recurrence. PMID- 11727627 TI - Results of preoperative ultrasound guided fine needle aspiration biopsy of solitary thyroid nodules as compared with the histology. A retrospective analysis of 538 patients. AB - AIM: The goal of this study was to assess the accuracy and limitations of ultrasound guided fine-needle aspiration biopsy (ug-FNAB) of solitary thyroid nodules. METHODS: The ug-FNAB results of 538 patients with solitary thyroid nodules, who afterwards underwent thyroid surgery, were compared retrospectively with the histology. Patients with multinodular goiter were excluded from the study. Ug-FNAB was performed on growing and/or hypoechoic and/or hypofunctional nodules. The ug-FNAB results were grouped as follows: group 1: malignant (n = 44); group 2: malignancy cannot be ruled out (n = 173); group 3: non-malignant (n = 296), group 4: inadequate (n = 25). RESULTS: When the cytological results of group 1 and group 2 were interpreted as being malignant and those of group 3 as being benign, sensitivity, specificity and accuracy of ug-FNAB were 96.7%, 65.8% and 69.5% respectively. The 62 thyroid carcinomas (TC) biopsied presented in 59 cases a suspicious or malignant cytology (95.2%). The smallest TC diagnosed by ug FNAB had a diameter of 0.5 cm and 36.4% of all papillary TC < or = 1 cm displayed stage pT4. The histology verified a TC in 18 cases out of the 173 ug-FNABs in group 2. Non-malignant ug-FNABs were confirmed by histology in 294 patients (99.3%) in group 3. In 4.65% of the ug-FNABs inadequate material was aspirated. CONCLUSION: Nodules with non-suspicious ug-FNAB results can be safely followed-up by sonography, as the cytological diagnoses were verified in more than 99% by histology. Papillary TC can be diagnosed with ug-FNAB very accurately. As stage pT4 was present in more than one third of patients with papillary TC < or = 1 cm, ug-FNAB is also recommended for thyroid nodules 0.5-1 cm in diameter located adjacent to the thyroid capsule. However, microfollicular proliferations remain the limitation of ug-FNAB, as the cytology cannot distinguish between benign adenoma and follicular TC. PMID- 11727628 TI - [Recovery test or immunoradiometric measurement of anti-thyroglobulin autoantibodies for interpretation of thyroglobulin determination in the follow-up of different thyroid carcinoma]. AB - The determination of thyroglobulin (Tg) in the follow-up of differentiated thyroid carcinomas (DTC), is routinely used in nuclear medicine, although some problems, like a disturbed recovery-test (RT) or autoantibodies to thyroglobulin (TgAb), are well known. But it is a controversial issue in literature, whether the determination of TgAb should be performed beside or instead of the RT. OBJECTIVE: The study compares the clinical value of the determination of both TgAb and RT with sensitive assays. METHODS: 356 patients (pts) were investigated. The results were compared to the concentration of Tg in the sera of the pts. 288 pts stayed tumor-free, the remaining 68 pts showed a recurrence (local and/or metastatic) of their DTC. We measured Tg (with RT) using an immunoradiometric assay (Tg-IRMA; SELco Tg; Fa. Medipan Diagnostica GmbH) and TgAb using a direct assay (CentAK anti-Tg; also from Fa. Medipan). RESULTS: The prevalence of TgAb, and of disturbed RT respectively, in the whole population of DTC-pts was 7.6%, in the subgroup of tumor-free pts 6.6%, and in the remaining pts with tumor recurrence 11.8%, respectively 2.0%, 1.7% and 2.9%. In a significantly higher percentage of pts with local/metastatic recurrence, both a positive TgAb (p < 0.001) and a disturbed RT (p < 0.05) were found. 7/68 pts with tumor-recurrence but Tg < 1 ng/ml showed positive TgAb, only 2/7 had a disturbed RT. In this group, no patient with Tg > 1 ng/ml demonstrated either positive TgAb or disturbed RT (p < 0.001 and p < 0.05). CONCLUSION: The determination of TgAb in the follow-up of DTC is necessary, because it supports a suspicion to tumor recurrence in pts with negative Tg. Also the RT is of great value because of a possibly High dose hook-effect. PMID- 11727629 TI - [An approach for comparative quantification of myocardial blood flow (O-15-H2O PET), perfusion (Tc-99m-tetrofosmin-SPECT) and metabolism (F-18-FDG-PET)]. AB - AIM: In the present study a new approach has been developed for comparative quantification of absolute myocardial blood flow (MBF), myocardial perfusion, and myocardial metabolism in short-axis slices. METHODS: 42 patients with severe CAD, referred for myocardial viability diagnostics, were studied consecutively with 0 15-H2O PET (H2O-PET) (twice), Tc-99m-Tetrofosmin SPECT (TT-SPECT) and F-18-FDG PET (FDG-PET). All data sets were reconstructed using attenuation correction and reoriented into short axis slices. Each heart was divided into three representative slices (base, midventricular, apex) and 18 ROIs were defined on the FDG PET images and transferred to the corresponding H2O-PET and TT-SPECT slices. TT-SPECT and FDG-PET data were normalized to the ROI showing maximum perfusion. MBF was calculated for all left-ventricular ROIs using a single compartment-model fitting the dynamic H2O-PET studies. Microsphere equivalent MBF (MBF_micr) was calculated by multiplying MBF and tissue-fraction, a parameter which was obtained by fitting the dynamic H2O-PET studies. To reduce influence of viability only well perfused areas (> 70% TT-SPECT) were used for comparative quantification. RESULTS: First and second mean global MBF values were 0.85 ml x min-1 x g-1 and 0.84 ml x min-1 x g-1, respectively, with a repeatability coefficient of 0.30 ml x min-1 x g-1. After sectorization mean MBF_micr was between 0.58 ml x min-1 x ml-1 and 0.68 ml x min-1 x ml-1 in well perfused areas. Corresponding TT-SPECT values ranged from 83% to 91%, and FDG-PET values from 91% to 103%. All procedures yielded higher values for the lateral than the septal regions. CONCLUSION: Comparative quantification of MBF, MBF_micr, TT-SPECT perfusion and FDG-PET metabolism can be done with the introduced method in short axis slices. The obtained values agree well with experimentally validated values of MBF and MBF_micr. PMID- 11727630 TI - [[F-18]FDG imaging of head and neck tumors: comparison of hybrid PET, dedicated PET and CT]. AB - AIM: Aim of the study was to evaluate [F-18]FDG imaging of head and neck tumors using a Hybrid-PET device of the 2nd or 3rd generation. Examinations were compared to dedicated PET and Spiral-CT. METHODS: 54 patients suffering from head and neck tumors were examined using dedicated PET and Hybrid-PET after injection of 185-350 MBq [F-18]FDG. Examinations were carried out on the dedicated PET first followed by a scan on the Hybrid-PET. Dedicated PET was acquired in 3D mode, Hybrid-PET was performed in list mode using an axial filter. Reconstruction of data was performed iteratively on both, dedicated PET and Hybrid-PET. All patients received a CT scan in multislice technique. All finding have been verified by the goldstandard histology or in case of negative histology by follow up. RESULTS: Using dedicated PET the primary or recurrent lesion was correctly diagnosed in 47/48 patients, using Hybrid-PET in 46/48 patients and using CT in 25/48 patients. Metastatic disease in cervical lymph nodes was diagnosed in 17/18 patients with dedicated PET, in 16/18 patients with Hybrid-PET and in 15/18 with CT. False positive results with regard to lymph node metastasis were seen with one patient for dedicated PET and Hybrid-PET, respectively, and with 18 patients for CT. In a total of 11 patients unknown metastatic lesions were seen with dedicated PET and with Hybrid-PET elsewhere in the body. Additional malignant disease other than the head and neck tumor was found in 4 patients. CONCLUSION: Using Hybrid-PET for [F-18]FDG imaging reveals a loss of sensitivity and specificity of about 1-5% as compared to dedicated PET in head and neck tumors. [F-18]FDG PET with both, dedicated PET and Hybrid-PET is superior to CT in the diagnosis of primary or recurrent lesions as well as in the assessment of lymph node involvement. PMID- 11727631 TI - [SAPHO syndrome--results of 2-phase bone scintigraphy]. PMID- 11727633 TI - Let's put "caring" back into healthcare: teaching staff to care. PMID- 11727634 TI - Role changes in mergers. PMID- 11727635 TI - Creating a research partnership: graduate nursing students and clinical nurses. PMID- 11727636 TI - A strategic vision for nursing practice. PMID- 11727637 TI - Hong Kong healthcare system and its challenges. PMID- 11727638 TI - Using organizational artifacts to influence change. PMID- 11727639 TI - Outcomes among orthopedic patients in skilled nursing facilities: does managed care make a difference? AB - The use of rehabilitative care has increased greatly. This study evaluates whether managed care affects health outcomes among Medicare orthopedic patients receiving rehabilitative treatments. Managed care versus fee-for-service patients had better outcomes at four months following discharge from skilled nursing facilities. It is important to address predictive factors, such as age, length of hospital stay, debilitation and social living arrangements, which can also influence health outcomes when planning rehabilitative treatment for older patients. PMID- 11727640 TI - Sexual harassment in healthcare: classification of harassers and rationalizations of sex-based harassment behavior. AB - This study identified how 16 Australian registered nurses classified sex-based harassers and explained their own behavior and the behavior of the harasser. A qualitative research design, relying on in-depth interviews, was used to collect the data. The study found that harassment is linked to gender roles and that the harassed are reluctant to blame the harasser--the harassed had "sound" rationalizations for harassment. Awareness of the interactional dynamics of self blame and these rationalizations will help nurse executives ensure a harassment free workplace. PMID- 11727641 TI - Attaining superior outcomes with joint replacement patients. AB - Wanting to improve the quality outcomes of their joint care program, a team approach was used to implement a program with integrated care across the continuum. The authors describe the joint care program and its outcomes that included a decrease in length of stay, fewer complications, improved functional status, higher discharge disposition to home, and lower variable costs per case. PMID- 11727642 TI - Coverage of adolescent substance use prevention in state frameworks for health education. AB - Ten secondary health education state curriculum frameworks were reviewed for their inclusion of 12 mediators commonly used to prevent adolescent substance use. Specific aims of the investigation were: a) to identify the extent to which the 12 mediators were found in each framework; and b) to identify those frameworks that included Alcohol, Tobacco, and Other Drugs (ATOD) sections and determine to what extent the 12 mediators were found in those sections. A panel of three researchers independently reviewed each framework. Beliefs about consequences, decision-making skills, and stress management skills were identified most often while commitment, lifestyle incongruence, and normative beliefs were identified least often. Among states that included ATOD sections, beliefs about consequences and resistance skills were the most commonly identified mediators. Commitment, goal setting, and normative beliefs were not identified in any ATOD sections. Research in prevention and implications for health education are discussed. PMID- 11727643 TI - A YRBS survey of youth risk behaviors at alternative high schools and mainstream high schools in Hong Kong. AB - In Hong Kong, prevocational schools serve as an alternative to mainstream schools to provide education with more emphasis on practical and technical subjects. In this paper, health-risk behaviors of prevocational school (PVS) students were identified, and comparisons of health-risk behaviors with or without adjusting the demographic factors from prevocational schools and mainstream schools were made. The PVS students were at higher risk for most categories of health-risk behaviors such as unintentional and intentional injuries, smoking, alcohol drinking, glue sniffing, inadequate physical activity, insufficient consumption of fresh fruits and vegetables, and early sexual activity with multiple partners. Female students of PVS reported higher prevalence of emotional problems and substance abuse. Findings suggest that the school environment is an influential factor on the lifestyle behavior of students. Comprehensive health education and intervention programs are needed for youth in Hong Kong prevocational schools. PMID- 11727644 TI - Primary grade teachers' knowledge and perceptions of head lice. AB - This study examined primary grade teachers' knowledge of head lice, perceived self-efficacy in dealing with head lice, and preferred resources of additional information on head lice. Survey data from a three-wave mailing (M = 292; 60%) found that teachers needed more knowledge regarding head lice and were significantly more knowledgeable as teaching experience increased. A plurality (46%) had high efficacy expectations scores regarding their ability to control the spread of head lice. Most (71%) primary teachers reported they received most of their head lice information from school nurses. Most (63%) said they would like more information on how to prevent head lice infestation, and they wanted the information in the form of videotapes (68%) and brochures (51%). Only 23% provided yearly instruction regarding head lice. PMID- 11727645 TI - A comparison of menstrual changes expected by pre-menarcheal adolescents and changes actually experienced by post-menarcheal adolescents in Mexico. AB - This survey explored para-menstrual changes that Mexican pre-menarcheal girls expect to experience and paramenstrual changes actually experienced by post menarcheal girls. The survey included 95 pre-menarcheal girls who reported what they expected to experience in the para-menstruum, and their answers were compared with para-menstrual changes reported by 98 post-menarcheal girls. There were more post-menarcheal than pre-menarcheal girls who reported negative physical changes, while more pre-menarcheal girls expected to experience negative changes related to cognitive-emotional expressions. Regarding positive para menstrual changes, despite the fact that hardly any of the girls named them, more post-menarcheal girls reported positive changes than pre-menarcheal girls who expected the changes. The dominant menstrual discourse concerning menstruation focused on para-menstrual negative changes, which can lead pre-menarcheal girls to anticipate their periods with fear and to overlook positive changes that may occur in the para-menstruum. PMID- 11727646 TI - Effectiveness of a health course at influencing preservice teachers' attitudes toward teaching health. AB - Research has shown that effective school health education depends on the effectiveness of health training for prospective teachers. This research tested the effectiveness of a university course in preparing preservice teachers in the area of school health. Education majors enrolled in the course "Drug and Health Issues for Educators" at Montana State University were administered a pretest/posttest questionnaire at the beginning and end of spring semester 2000 to gather data for testing five hypotheses. Analyses were conducted on 119-paired samples. The course appeared effective at influencing how a preservice teacher values health education in the school, their intentions to include and confidence in including health in their teaching, and their ability to provide a student with information on personal health issues. PMID- 11727647 TI - Assessment strategies for school-based mental health counseling. PMID- 11727649 TI - Statewide consortium grapples with "quality care" issues. Minnesota sets the stage for consistent use of best medical practices. PMID- 11727648 TI - Vulvar health article heightens awareness. PMID- 11727650 TI - Solving a severe asthma episode. What is your diagnosis? Churg-Strauss syndrome. PMID- 11727651 TI - Hypertension syndrome and cardiovascular events. High blood pressure is only one risk factor. AB - It is becoming increasingly clear that high blood pressure is not the sole cause of the high cardiovascular morbidity and mortality rates associated with hypertension. Reduction of blood pressure is of utmost importance, but many other factors contribute significantly to the risk of adverse cardiovascular events and death. In this article, Dr Glasser reviews hypertension as a syndrome, emphasizing therapy to improve blood pressure control, increase arterial compliance, and inhibit or reverse vascular remodeling. PMID- 11727652 TI - Initial antihypertensive therapy. What are the current drugs of choice? AB - The goal of antihypertensive treatment, in addition to lowering blood pressure, is to reduce the risk of cardiovascular events. Until recently, however, only conventional treatment with diuretics and beta-blockers had been studied in terms of cardiovascular end points. In this article, Dr Yeun reviews the results of recent trials comparing these agents with other classes of antihypertensive drugs. She examines the confounding elements in the trials, provides an interpretation of study results, and suggests a practical approach to initial treatment of uncomplicated hypertension. PMID- 11727653 TI - Posttraumatic stress disorder. Safe, effective management in the primary care setting. AB - PTSD is a severe and chronic psychiatric condition. Because of the complex symptoms, no single treatment is effective. Pharmacologic treatment is an integral component of a multidisciplinary approach that addresses the psychological, social, and spiritual dimensions of PTSD. Psychopharmacologic agents provide symptomatic relief and are beneficial in managing other comorbid psychiatric conditions. Primary care physicians can base treatment decisions on the patient's prevailing symptoms. Specialized psychiatric consultation may be needed in difficult and complicated cases. PMID- 11727654 TI - Tympanic membrane perforation in adults. How to manage, when to refer. AB - Tympanic membrane perforation is a common and potentially serious condition. One of the most important aspects of primary care for perforations is deciding which patients need to be seen by an otolaryngologist and how urgently they need to be referred. There are several indications for surgical repair, but most cases can be managed with conservative care and require no referral. PMID- 11727655 TI - Ten pivotal questions about diabetic ketoacidosis. Answers that clarify new concepts in treatment. AB - Nearly all physicians have cared for patients with diabetic ketoacidosis sometime during their training or have encountered patients with hyperglycemia and ketonuria in their office practice. In the last decade, many studies have challenged the traditional concepts about diabetic ketoacidosis treatment, resulting in sometimes confusing recommendations. In this article, Drs Carroll and Schade answer 10 frequently asked questions about the diagnosis and treatment of diabetic ketoacidosis and discuss related hospitalization issues. PMID- 11727656 TI - Patient notes. Post-traumatic stress disorder. PMID- 11727657 TI - Finding the e in healthcare. PMID- 11727658 TI - Brave new imaging world. PMID- 11727659 TI - From behind closed doors. PMID- 11727660 TI - Healthcare innovations in technology systems. Team operations. PMID- 11727661 TI - Engineering data exchange. AB - Mount Clemens General Hospsital, Mount Clemens, Mich. PROBLEM: performance anomalies due to multiple interfaces with separate systems. SOLUTION: implementation of an interface engine. RESULTS: improved management of data exchange among disparate systems. KEYS TO SUCCESS: up-front studies of vendor offerings, potential problems, and long-term needs. PMID- 11727662 TI - Top five priorities among CEOs. PMID- 11727663 TI - [Hematopoietic growth factors in prevention and therapy of infectious complications in premature and newborn infants]. AB - Bacterial sepsis continues to be an important cause of morbidity and mortality in neonates. Neutropenia in the newborn, as a result of decreased bone marrow neutrophil storage pool reserves and myeloid committed progenitor cells, increases the risk of sepsis and is associated with a poor prognosis. The hematopoietic colony-stimulating factors G-CSF (granulocyte colony stimulating factor) and GM-CSF (granulocyte-macrophage colony stimulating factor) increase the number of circulating neutrophil number by stimulating neutrophil precursors proliferation. In a number of clinical trials in very low birthweight neonates and in neonates with preeclampsia-associated neonatal neutropenia, both hematopoietic growth factors significantly increased the circulating absolute neutrophil count. However, no larger study showed that prophylactic G-CSF or GM CSF treatment resulted in a reduction of infectious complications or in an improved overall survival. Similar results were seen in studies evaluating G-CSF and GM-CSF as intervention therapy in septic neonates. Therefore, experts do not recommend the routine use of the expensive growth factors in preterm and term neonates. However, prospective clinical trials are still needed to evaluate whether specific treatment groups will benefit from the use of G-CSF or GM-CSF. In this regard, efforts must be directed at better defining the endpoints and in particular assigning value to reduction in treatment of possible infectious complications, such as days in hospital, antibiotic usage and costs. In addition, randomized studies are required to evaluate the proper dosage and duration of therapy, which most likely will vary between groups of patients. PMID- 11727664 TI - [Fetal surgery for severe congenital abnormalities]. AB - BACKGROUND: Over the past two decades the diagnosis of life threatening congenital malformations has evolved rapidly. Sophisticated and powerful new imaging and sampling techniques have stripped the veil of mystery from the once secretive fetus. Early detection and close follow-up of the fetus with congenital malformations have allowed us to define their natural history, determine the clinical features that affect clinical outcome, and plan management approaches to improve prognosis. Fetal surgical intervention is the logical culmination of the progress in fetal diagnosis. The purpose of this article is to describe the current techniques and recent advances in prenatal diagnosis and fetal intervention of severe congenital malformation. MATERIAL AND METHODS: A complete review of the literature and our own experience concerning fetal surgery was performed. RESULTS: Although most prenatally diagnosed malformations are best managed by appropriate medical and surgical therapy after maternal transport and planned delivery at a tertiary care center, an expanding number of simple anatomical abnormalities with predictable, lethal consequences have been successfully corrected before birth. A malformation amenable to prenatal surgical intervention must fulfill a number of conditions. It must be severe enough to warrant the risks associated with in utero treatment and must be reliably detectable before birth. Additionally, the pathophysiology must be reversible by fetal surgery, significantly improving the prognosis over post-natal treatment. Many technical intricacies of open fetal surgery have been solved, but pre-term labor and premature rupture of membranes remain a omnipresent risks to both the mother and the fetus. To reduce maternal morbidity and the risk of prematurity we developed minimally invasive techniques to treat the fetus prenatally. Current indications of fetal surgery include the treatment of congenital diaphragmatic hernia, cystic adenomatoid malformation of the lung, sacrococcygeal teratoma, obstructive uropathy, twin-to-twin-transfusion-syndrome and myelomeningocele. Minimally invasive surgical techniques (FETENDO) have significantly lessened the incidence of preterm labor and promise to extend the indications for fetal surgical intervention. CONCLUSIONS: Fetal surgical therapy for severe congenital malformations may improve the outcome of selected patients. The development of FETENDO will in all probability reduce the importance of open fetal surgery in the future. PMID- 11727665 TI - [Nitroglycerin patch for tocolysis--a prospective randomized comparison with fenoterol by infusion]. AB - OBJECTIVE: To evaluate tocolytic efficacy of transdermal glyceryl trinitrate (GTN) in comparison to fenoterol per infusionem in a prospective randomized multicenter study. PATIENTS AND METHODS: 50 pregnant women between 27 and 35 weeks of gestation with preterm labour were treated with either GTN patches (0.4 0.8 mg/h) or fenoterol per infusionem (60-120 micrograms/h) up to stop of contractions or 35 weeks in maximum. The primary outcomes were the prolongation of gestation by 48 h, 7 days or up to 37 weeks of gestation as well as the neonatal outcome. The progression of cervical ripening and maternal side effects during tocolysis were assessed as secondary outcome criteria. RESULTS: There was no difference in successful tocolysis for 48 h and 7 days in both groups, whereas significantly more women passed 37 weeks after GTN therapy. So mean duration of pregnancy, birth weight and height were greater, whereas transfer into neonatal care unit was significantly rare after GTN. There were no differences in neonatal outcome and progression of cervical ripening during tocolysis. Maternal side effects during GTN were fewer and weaker compared with fenoterol. Circa 70% of GTN treated women had a headache temporary, whereas more than 90% of the patients with fenoterol suffered from tachycardia and tremor. CONCLUSIONS: Tocolytic efficacy of transdermal GTN was at least equivalent to the established beta mimetic therapy with fenoterol. Because of the lower preterm delivery rate transfer into neonatal care for control was significantly rarer after GTN with equally good neonatal outcome in both groups. Beside the headache transdermal GTN therapy had lower maternal side effects in comparison to fenoterol. PMID- 11727666 TI - [Prune belly syndrome and congenital kidney tumors]. AB - BACKGROUND: High end sonography allows the prenatal localization of the kidneys and the corresponding urine drainage system as early as 10-13 weeks of gestation. In mid second trimester, the voiding and filling of the urinary bladder can be demonstrated by ultrasound. Obstructions are the most common abnormalities of the urogenital tract. Though less frequent in incidence, more complex sequences of anomalies such as Prune Belly Syndrome or Megacystis-Microcolon-Intestinal Hypoperistalsis-Syndrome (MMIHS) can also be detected in early gestational age. MATERIALS AND METHODS: Pathogenesis, prenatal diagnosis, pre- and postnatal treatment options and prognosis are discussed. RESULTS AND DISCUSSION: The same risk-adapted procedures aimed to protect the fetal urinary excretory function known in the therapeutic regimen of obstructive uropathy are available as treatment options. These range from non-invasive ultrasound for diagnosis and surveillance to needle procedures or even endoscopic interventions. Another rare entity of renal abnormalities are congenital neoplasm's--megaloblastic nephroma, nephroblastoma and neuroblastoma. CONCLUSION: Prognosis and obstetrical management are to be determined individually for each patient. PMID- 11727667 TI - [Fetomaternal macrotransfusion after attempted external version]. AB - BACKGROUND: External cephalic version is an alternative to both vaginal breech delivery and to caesarean section. The objective of this study was to summarize potential complications of external cephalic version. Which means are available for early detection and treatment of these complications? CASE REPORT: A 28-year old 1/0 with breech presentation was considered for external cephalic version at the 38th week of gestation. As three version attempts failed, the patient was discharged after normal fetal heart tone (FHT) registration and normal ultrasonographic findings on the next day. Caesarean section was planned six days later. On the day of admission, FHT registration revealed a decreased beat-to beat variability and a sinusoidal baseline. Doppler-flow indices of the A. umbilicalis and A. cerebri media were normal, although middle cerebral artery peak systolic velocity was increased. Rapid caesarean section was performed, and an anemic baby (hemoglobin 3.4 g/dl) was born. After transfusion of 100 ml red blood cells, further development of the newborn was normal. DISCUSSION AND CONCLUSION: Fetomaternal macrotransfusion may be a rare complication of external cephalic version, occurring even several days after the mechanical manipulation. FHT registrations and ultrasonographic doppler flow measurements performed periodically unit birth are necessary to detect such complications early. The Kleihauer-Betke test is the method of choice to diagnose fetomaternal macrotransfusion. PMID- 11727668 TI - Hyperpolarised gases in magnetic resonance: a new tool for functional imaging of the lung. AB - In magnetic resonance imaging (MRI), nuclear spins are the source of the image signal. In the lung, low-proton spin density in alveolar gas and abundant gas tissue interfaces substantially impair conventional native 1H-MRI. Spin polarisation can be increased in two non-radioactive noble gas isotopes, 3He and 129Xe, by exposure to polarised laser light. When inhaled, such "magnetized" gases provide high-intensity MR images of the pulmonary airspaces. Thus, hyperpolarised gas (HPG) MRI opens up new routes to a) morphologic imaging of airways and alveolar spaces, and b) analysis of the intrapulmonary distribution of inhaled aliquots of these tracer gases; c) diffusion-sensitive MRI-techniques allow mapping of the "apparent diffusion coefficient" (ADC) of 3He within lung airspaces, where ADC is physically related to local bronchoalveolar dimensions; d) also, 3He magnetisation decays in an oxygen-containing atmosphere at a rate proportional to ambient PO2. This property allows image-based determination of regional broncho-alveolar PO2 and its decrease during a breathhold. Currently, these modalities of functional lung imaging are being assessed by several European and American research groups in animal models, human volunteers and patients. First results show good imaging quality with excellent spatial and unprecedented temporal resolution, and attest to the reproducibility, feasibility and safety of the technique. Regionally impaired ventilation of both structural and functional origin is detected with high sensitivity, e.g. in smokers, asthmatics, patients with COPD or after lung transplantation. Studies into regional ADC and PO2 measurement demonstrate good agreement with reference methods and physiological predictions. The present limitations of HPG-MRI include the HPG production rate and the US and EU health authorities' still pending final approval for clinical use. PMID- 11727669 TI - Exercise training in chronic heart failure: why, when and how. AB - The management of patients with chronic congestive heart failure has changed considerably during the last decade. Until recently, restriction of physical activity was recommended for patients with chronic heart failure. However, the knowledge that training influences largely the periphery rather than the heart itself has led to a dramatic change in the approach toward training in patients with chronic heart failure. Why to train patients with chronic heart failure: Training increases exercise tolerance by an average of 20% in chronic heart failure regardless of etiology (ischemic or non-ischemic cardiomyopathy) or severity of left ventricular dysfunction. Available data, while limited, demonstrate that increases in exercise capacity are paralleled by an improvement in quality of life. Studies have consistently demonstrated that training has no deleterious effect on central haemodynamics, left ventricular remodeling, systolic or diastolic function, or myocardial metabolism. At present, there are insufficient data to determine the effect of training on prognosis, but trials are currently underway to address this. When to train patients with chronic heart failure: Exercise training should be performed only with the patients that have been in a stable clinical condition for a period of at least 3-4 weeks. Clinical stability is defined as no change in symptoms, weight, drug regimen, or NYHA class over this period. How to train patients with chronic heart failure: Initially, the program should be supervised for a period of 2 to 4 weeks; home based programs are usually appropriate thereafter. Activities that can be maintained for a lifetime should be encouraged, and the focus should be on aerobic-type activities. The intensity level should be targeted to about 50%-70% of peak VO2 and/or Borg ratings of 12-14 ("walk and talk"). PMID- 11727670 TI - Contraception and sexuality after termination of pregnancy: a comparison between Lugano and Geneva. AB - OBJECTIVE: To evaluate the impact of a voluntary termination of pregnancy (TOP) on contraceptive practice and sexual well-being in women. STUDY DESIGN AND METHODS: Comparative evaluation of a retrospective study conducted in Lugano in 1995 with 85 women interviewed 3-6 months post-TOP, and a prospective study carried out in Geneva in 1998 with 103 women interviewed pre-TOP and 6 months post-TOP. In both studies TOP was performed during the first 12 weeks of pregnancy by vacuum aspiration and curettage. RESULTS: Oral contraceptive use in the Lugano group increased from 21% pre-TOP to 47% post-TOP, and in the Geneva group from 19% pre-TOP to 69% post-TOP. Condom use decreased in both groups. Those practising unprotected intercourse also decreased by half (around one in five). No effect on sexual well-being was reported by almost two-thirds of both groups. CONCLUSIONS: Despite large differences between these two populations, the two studies show a similar evolution. The majority of women appeared to be able to cope well with a TOP. About one third presented sexual dysfunction. PMID- 11727671 TI - Motivation for smoking cessation: what role do doctors play? AB - PRINCIPLES: Cigarette smoking causes an estimated 13% of all deaths in Switzerland. Though most smokers will eventually become ex-smokers of their own volition, physicians play an important role in accelerating the process of quitting among smoking patients. Even brief advice from physicians is effective in doing so. The purpose of this study was to investigate which smokers were asked about their smoking habits, and how often, whether they received advice to quit, and how this correlates with the patient's desire to quit. METHODS: Telephone interviews were carried out with a random sample of smokers and ex smokers from the German-speaking Swiss population (n = 993). We collected information on personal characteristics, smoking habits, and recall of physicians' advice. Data was analysed descriptively and by logistic regression. RESULTS: 88% recalled being asked by a doctor about their smoking habits. In contrast, only 34% of smoking patients recalled being advised to stop. Women, older people and those in poor subjective health were asked more frequently. Heavier smokers and those in poor subjective health were advised more frequently. Current smokers more frequently express the desire to quit if they are heavier smokers and have been advised to quit by their physician, compared with those who have not received such advice. CONCLUSIONS: Similarly to the international findings, smoking patients in Switzerland receive brief advice with insufficient frequency. Action should therefore be taken to encourage health professionals not only to question all smoking patients but to advise and motivate them to quit smoking. PMID- 11727672 TI - Violence in Bern. PMID- 11727673 TI - [Occurrence of scrapie in Switzerland: an anonymous cross-sectional study]. AB - Scrapie is a transmissible spongiform encephalopathy of sheep and goats, which leads to neurological signs and ends with the death of the infected animal. In Switzerland only a few cases were reported since 1982. In 1999, a questionnaire survey was conducted to increase information about neurological diseases in sheep and goat breeding farms. The aim was to estimate the frequency of neurological disorders including Scrapie and to increase the disease awareness of sheep and goat breeders. The main goal of the study was to increase the reporting of suspect cases of Scrapie and its differential diagnoses. Out of a database of all registered farms with at least five breeding sheep or breeding goats a random sample of 4711 was drawn to which anonymous questionnaires were sent. The return rate was 36%. In a parallel study, 150 voluntarily participants were interviewed using the same questionnaire. The results of both parts of the survey coincided. For the years 1997 and 1998, on average 1.0% of the breeding sheep and breeding goats showed neurological signs. 7.7% of the breeders indicated to have observed animals with neurological symptoms in their flock. At the population level, a total of 1954-2336 animals with neurological signs are observed in 691-929 flocks. The minority is reported to the veterinary authorities. PMID- 11727674 TI - [Clinical findings and diagnostic procedure in a dairy sheep with visna]. AB - The goal of this report was to describe the clinical signs and diagnosis of Visna in a seven-year-old East Friesian milk sheep. A striking feature was that the ewe's behaviour changed frequently. At one time, the ewe was somnolent. A few minutes later, the sheep was alert and eating hay. The ewe was thin. It had a slight head tilt and a severe generalised ataxia. Based on the neurological symptoms and chronic weight loss, a tentative diagnosis of visna was made. Serological testing for maedi-visna was positive, and the ewe was euthanised. A postmortem examination was performed, and lung and brain samples were collected aseptically. Cell cultures from these organs were positive for viral enzymatic reverse transcriptase and for maedi-visna RNA. PMID- 11727675 TI - [Myxomatosis in the rabbit]. PMID- 11727676 TI - On postdoctoral research fellowships in mental health nursing. PMID- 11727677 TI - Is coronary atherosclerosis an infectious disease? PMID- 11727678 TI - Silent myocardial ischemia: is it a clinical entity that requires therapy? PMID- 11727679 TI - Nutriceuticals as treatments for cardiovascular disease. PMID- 11727680 TI - The clinical trial under fire. PMID- 11727681 TI - The past is past. PMID- 11727682 TI - Why not the best? PMID- 11727683 TI - Electrocardiographic telemetry: determining its value in emergency medical services. PMID- 11727684 TI - High-yield selection criteria for cranial computed tomography after acute trauma. AB - OBJECTIVE: To develop and prospectively evaluate criteria for selecting head injured patients requiring cranial CT. METHODS: A two-phased prospective observational study design was implemented at a university ED. Physicians completed a form with 51 variables for 540 patients in Phase I. Ten high-yield variables were identified and prospectively tested on 273 patients in Phase II. Prediction rule performance for identification of patients with abnormal CT scans was determined. RESULTS: The combined criteria had a sensitivity of 97% for CT scan abnormalities in Phase I; sensitivity was 96% in Phase II. Negative predictive values were 97% and 94%, respectively. Prevalence of disease in Phase I was 17%; prevalence in Phase II was 16%. Had the Phase I criteria been implemented during Phase II, 43 of 273 patients (16%) would not have been scanned, including two patients with positive results who did not require operative intervention. CONCLUSIONS: Fifty-one clinical variables from head injured patients were narrowed to ten of statistical significance and consistent interpretation for prospective evaluation. Patients with none of these criteria were found to be at low risk of having sustained significant head injury. PMID- 11727685 TI - The effects of warming and buffering on pain of infiltration of lidocaine. AB - OBJECTIVES: To investigate the effects of warming vs buffering, and warming with buffering, on the pain of lidocaine infiltration. METHODS: A randomized, double blind clinical trial was conducted using volunteers aged 18 years or more and without an allergy to lidocaine. The study consisted of two parts, each comparing two solutions. The solutions for Part I were warm (38.9 degrees C; 102 degrees F) plain lidocaine and room-temperature buffered lidocaine. Warm buffered lidocaine and room-temperature buffered lidocaine were used for Part II. The subjects received two standardized 0.5-mL intradermal injections, one study solution in each forearm. Immediately after each injection, pain was assessed using a 100-mm visual analog pain scale. Pain scores were analyzed by the sign test, with significance defined as p < 0.05. RESULTS: Part I (n = 10): Nine subjects reported room-temperature buffered lidocaine to be less painful than warm plain lidocaine. Mean pain scores were 28 mm lower for room-temperature buffered lidocaine than they were for warm plain lidocaine (p < 0.01). Part II (n = 24): Eleven subjects found warm buffered lidocaine to be the least painful, 11 reported room-temperature buffered lidocaine to be the least painful, and two reported no difference. A mean pain score difference of 2.1 mm favoring warm buffered lidocaine was not statistically significant. Part II had a power of 80% to detect a 10-mm difference between the two solutions at alpha = 0.05. CONCLUSION: To reduce the pain of lidocane infiltration, buffering is more effective than warming. Warming does not enhance buffering. PMID- 11727686 TI - Efficacy of tissue glue for laceration repair in an animal model. AB - OBJECTIVE: To compare the efficacy of butyl-2-cyanoacrylate tissue glue (TG) for the repair of skin lacerations in rats with the efficacy of standard closure with sutures. METHODS: In a prospective study, eight rats were anesthetized and an 8 cm dorsal incision was made on each side of the midline. One wound was closed with a single layer of interrupted 5-0 Prolene suture and one by application of TG. The time required to close each wound was recorded. Sutures were removed at seven days; the TG was allowed to fall off spontaneously. The animals were sacrificed after 20 days and the wounds were judged for cosmetic outcome. Four I x 3-cm strips of skin were excised from each wound; one strip for histologic analysis and three for load extension testing using a tensiometer. Specimens were loaded to wound failure while displacement (D) and energy absorption (EA) were recorded. The paired t-test was used for comparisons and reported as mean +/- SE. No significant difference between TG and suture was found in D (6.5 +/- 0.4 vs 5.2 (1.4 mm), EA (0.18 +/- 0.01 vs 0.17 +/- 0.03 kg x mm/cm2), or histologic features. The closure time was significantly less using TG (66 +/- 5 vs 401 +/- 17 sec; p < 0.0001). CONCLUSIONS: 1) Sutures and TG in rat skin repair result in similar wound strengths (EA), amounts of stretch (D), and histologic feasures. 2) Wound closure is accomplished much more rapidly with TG. PMID- 11727687 TI - Association of drug therapy with survival in cardiac arrest: limited role of advanced cardiac life support drugs. AB - OBJECTIVE: To generate hypotheses regarding the association of standard Advanced Cardiac Life Support (ACLS) drugs with human cardiac arrest survival. METHODS: This observational cohort study was conducted over a two-year period in the wards, intensive care units, and EDs of two tertiary care hospitals. Included werc adult patients who suffered cardiac arrest either inside or outside the hospital and who required epinephrine according to standard ACLS guidelines. Six standard ACLS drugs (given while CPR was in progress) were assessed for association with survival from resuscitation to one hour and to hospital discharge by univariate and multivariate logistic regression analyses. RESULTS: In the 529 patients studied, initial cardiac rhythm had no impact on the association between drug administration and survival. The time of drug administration (quartile of ACLS period) was associated with resuscitation for atropine (p < 0.05) and lidocaine (p < 0.01). The odds ratios (95% CIs) for successful resuscitation, after multivariate adjustment for potential confounders, were: a respiratory initiating cause, 3.7 (2.1 -6.4); each 5-minute increase in CPR-ACLS interval, 0.5 (0.4-0.7); each 5-minute duration of ACLS. 0.9 (()1.8- 1.0; atropine, 1.2 (1.0-1.3); bretylium. (0.4 (0.1-1.1); calcium 0.8 (0.2 2.4); lidocaine, 0.9 (0.7-1.1); procainamide. 21.0 (5.2-84.0) d sodium bicarbonate 1.2 (1.0-1.6). All other potential confounding variables entered into the model were not significantly associated with resuscitation. CONCLUSION: Initiating cause of arrest, time to ACLS, and duration of ACLS were important correlates of survival. Other than procainaimide, standard ACLS drugs had relatively little association with survival, but timing of administration may be an important factor. Further research using definitive large randomized controlled trials is warranted to assess the role of drug therapy in improving cardiac arrest survival. PMID- 11727688 TI - Physician medical direction and advanced life support in the United States. AB - OBJECTIVE: To determine the specialty training and responsibilities of urban U.S. emergency medical service (EMS) medical directors how these factors relate to the type of service involved (fire, hospital, private, municipal). METHODS: A single mailed survey of training officers--field supervisors of 211 urban advanced life support (ALS) services in the United States. The survey also requested information about medications carried, approved procedures. and who set these standards. Respondents also rated the EMS medical director's involvement in various activities (quality assurance, administrative, executive, run reviews, and in-service/education). RESULTS: Eighty-five percent (n = 179) of the forms were returned. with 165 (78%) usable. The physician EMS medical directors were primarily trained in emergency medicine (77%) and were paiid (75%) for EMS responsibilities. The number of medications carried and the number of approved procedures were not related to either the number of hours the physicians commit weekly to the EMS service or their degree of involvement in ALS activities. The physician EMS medical directors were most often involved in quality assurance and education and were less likely to devote time to executive or other administrative functions of ALS units, with the exception of fire-based EMS physician medical directors, who contributed significantly to executive and administrative functions (p < 0.05). Overall practice standards were established by the medical director (46%), the state department of health (24%), and local/regional health authorities (23%). CONCLUSIONS: EMS training officers believe that urban ALS medical directors in the United States primarily provide quality assurance and educational support. With the exception of fire-based EMS systems. physicians appear to have limited involvement in other EMS administrative and executive functions. PMID- 11727689 TI - On-line telemetry: prospective assessment of accuracy in an all-volunteer emergency medical service system. AB - OBJECTIVE: To evaluate the need for on-line telemetry control in an all volunteer, predominantly advanced emergency medical technician (A-EMT) ambulance system. METHODS: Emergency medical service (EMS) advanced life support (ALS) providers were asked to transmit the ECG rhythms of monitored patients over a six month period in 1993. The ECG rhythm interpretations of volunteer EMS personnel were compared with those of the on-line medical control physician. All discordant readings were reviewed by a panel of physicians to decide whether the misdiagnosis would have resulted in treatment aberrations had transmission been unavailable. RESULTS: Patients were monitored and rhythms were transmitted in 1,825 cases. 1,642 of 1,825 rhythms were correctly interpreted by the EMS providers (90%; 95% CI 89-91%). The accuracy of the EMS providers was dependent on the patient's rhythm (chi-square, p < 0.00001), the chief complaint (chi square, p = 0.0001), and the provider's level of training (chi-square, p = 0.02). Correct ECG rhythm interpretations were more common when the out-of-hospital interpretation was sinus rhythm (95%), ventricular fibrillation (87%), paced rhythm (94%), or agonal rhythm (96%). The EMS providers were frequently incorrect when the out-of-hospital rhythm interpretation was atrial fibrillation/flutter (71%), supraventricular tachycardia (46%), ventricular tachycardia (59%), or atrioventricular block (50%). Of the 183 discordant cases, 124 (68%) involved missing a diagnosis of, or incorrectly diagnosing, atrial fibrillation/flutter. Review of the discordant readings identified 11 cases that could have resulted in treatment errors had the rhythms not been transmitted, one of which might have resulted in an adverse outcome. CONCLUSIONS: In this all-volunteer, predominantly A-EMT ALS system, patients with a field interpretation of a sinus rhythm do not require ECG rhythm transmission. Field interpretations of atrial fibrillation/flutter, supraventricular tachycardia, ventricular tachycardia, and atrioventricular blocks are frequently incorrect and should continue to be transmitted. PMID- 11727690 TI - Effect of nitrous oxide analgesia on pneumothorax. AB - OBJECTIVE: To address whether spontaneous inhalation of 50% nitrous oxide (N2O) for up to 30 minutes adversely affects swine with pneumothorax (PNTX). METHODS: Five fasted, female swine (13-17 kg) were sedated with IV ketamine, intubated, and allowed to spontaneously inhale either 50% N2O or room air (RA) in a sequential crossover design in which each animal served as its own RA control. Small (group 1, 150 mL), medium (group II, 300 mL), and large (group III, 500 mL) PNTXs were created by instilling air into the left pleural space via an 18-Fr three-way Foley catheter. Changes in PNTX volume, heart rate (HR), central venous pressure (CVP), blood pressure (BP), ECG. and arterial blood gas (ABG) parameters were recorded in separate 10- and 30-minute trials. A 15-minute washout period was given between each trial. Data were analyzed using repeated-measures analysis of variance with post-hoc Tukey's tests. RESULTS: No significant increase in PNTX size was seen during the 10-minute trials. In the 30-minute trials, a statistically significant increase in absolute PNTX size was seen with N2O compared with RA for group I (44 +/- 17 vs 16 +/- 5 mL, p = 0.02), group II (61 +/- 21 vs 26 +/- 4 mL, p = 0.01), and group III (62 +/- 32 vs 40 +/- 5 mL, p = 0.06). No difference in HR, CVP, MAP, ABGs, or ECG were observed between the N2O and RA trials for any size PNTX. No animal developed hemodynamic signs of tension pneumothorax. CONCLUSION: In this model, spontaneous inhalation of 50% N2O for up to 30 minutes is associated with little risk of hemodynamic or respiratory compromise. Although PNTX size increases with 50% N2O use, the magnitude observed in this animal model is less than previously reported. PMID- 11727691 TI - Exertional heat stroke in a young woman: gender differences in response to thermal stress. AB - Exertional heat stroke (EHS) is an acute life-threatening emergency that necessitates the immediate institution of cooling measures. Reported here is a case of EHS in a nonacclimatized young woman who was undergoing strenuous exercise. The patient developed many of the characteristic features of EHS, including central nervous system disturbances, lactic acidosis, rhabdomyolysis, coagulopathy, and abnormal myocardial conduction. While EHS is relatively common in young men, the condition is rare in women. This case presentation addresses gender differences in the response to the thermal stress of intense physical activity. PMID- 11727692 TI - Unilateral facial swelling and fever. PMID- 11727693 TI - Falsely elevated serum digoxin levels secondary to endogenous digoxin-like immunoreactive substances. AB - Endogenous digoxin-like immunoreactive substances (DLIS) are produced by the human body and can be significantly elevated in specific clinical conditions. Commercially available digoxin assays do not have the specificity to fully distinguish DLIS from exogenous digoxin, though DLIS do not possess the same therapeutic properties as do the drug. The resultant artificial elevation of a reported digoxin level or a completely factitious level in a patient not taking the drug can have clinically significant consequences. PMID- 11727694 TI - Factors influencing applicants' match rank order. PMID- 11727695 TI - Moonlighting and student debt. PMID- 11727696 TI - ST-segment elevation as a discriminator in cocaine-associated chest pain. PMID- 11727697 TI - Complementary feeding: study on prevalence of food intake in two health centers of Sao Paulo city. AB - The infant feeding practices in the first year of life are of fundamental importance for their growth and development. This study was carried out aiming at checking on the prevalence of food intake by the infants during their first year of life. One-hundred-and-seventy-five children aged up to one year, attended to in two Health Centers of Sao Paulo city, Brazil, participated in this study. Their feeding practices, obtained through the status quo approach, were analyzed through multiple logistic regression models, using curves of prevalence for complementary food consumption. Fruit was the first solid food to be part of the infant diet, followed by vegetables, cereal, meat and/or eggs and, beans. Animal protein-containing source foods (meat and eggs) entered the diet much later, being consumed by practically all children only at the end of their first year of life. The early introduction of complementary foods into the infant diet was made evident. The introduction of solid foods to complement breastfeeding is started with low-calorie density foods, in disagreement with the recommendations for Brazilian children. Results of this paper disclose a need for having programmatic actions in health education being carried out, in special those regarding exclusive breastfeeding promotion and orientation on the adequate introduction of complementary feeding. PMID- 11727698 TI - Responding effectively when one employee accuses another of harassment. PMID- 11727699 TI - Disease management without disease managers. PMID- 11727700 TI - Policy and procedure manual: guidelines for disclosure and discussion of conditions and events with patients, families and guardians. PMID- 11727701 TI - Abstracts of the 2nd International Meeting on Antimicrobial Chemotherapy in Clinical Practice. Portofino, Italy, October 21-24, 2001. PMID- 11727702 TI - German and Austrian Society of Hematology and Oncology annual meeting. Mannheim, Germany, September 30 - October 3, 2001. Abstracts. PMID- 11727703 TI - Author index volume 1 (1974) to volume 50 (1998). PMID- 11727706 TI - Annual Meeting of the American Epilepsy Society. Philadelphia, Pennsylvania, USA. November 30-December 5, 2001. Abstracts. PMID- 11727707 TI - Abstracts of the 4th Biennial Congress of the European Society for Sexual and Impotence Research. 30 September-3 October 2001. Rome, Italy. PMID- 11727705 TI - [A turning point in the knowledge of the structure-function-activity relations of elastin]. AB - In this review are presented the last new results of our research group dealing with the molecular structures (atomic level) of tropoelastin, elastin and elastin derived peptides studied by using essentially methods of bioinformatics (theoretical predictions and molecular modelling) linked to experimental circular dichroism spectroscopic studies. We already had characterized both the local secondary structure and some parts of the tertiary structure of the tropoelastin and elastin molecules (human, bovine...), by using either theoretical predictions (local secondary structure, linear epitopes...) and/or experimental data (optical spectroscopic methods: Raman scattering, infrared absorption, circular dichroism). Except the cross-linking regions which are in helical conformations, the whole tropoelastin structure displays a lot of beta-reverse turns which usually belong to irregular structures in proteins. These turns play a key role in other regularly structures orientation (alpha-helix, beta-strand), thus they are very important in the native protein 3D architecture. It is particularly true for human tropoelastin, because its sequence is rich in glycines and prolines, and these residues are frequently met in beta-turns (a beta-turn is made of four consecutive residues which are stabilized by an hydrogen bond). Several types of beta-turns can be defined with the dihedral angles values phi and psi of the two central residues. Thus, by using a very recent updated set of propensities for the amino acid residues to belong to given types of reverse beta-turns (extracted from a reference set of known 3-D structures of globular proteins), we have determined, (by using our home made software COUDES), for all possible tetrapeptides of the human tropoelastin sequence, the distribution and the characterization of the possible type of turns. Thus, it is shown that the locations and/or the types of these reverse beta-turns reveal a regularity and are not all random. This confirms our hypothesis that intra-molecular elasticity of tropoelastin could be explained by the possibility of transitions between conformations involving short beta-strands and beta-turns. This result is of great interest in the construction (by using molecular biology) of elastic biomaterials derived from the elastin sequence (particularly, the elastin derived peptides corresponding to the sequence exon 21--(exon 24--exon 24...). Our study permit also to predict the conformations of specific elastin derived peptides which could have interesting biological activity. Peptides resulting from the degradation of elastin, the insoluble polymer of tropoelastin and responsible for the elasticity of vertebrate tissues, can induce biological effects and notably the regulation of matrix metalloproteinases (MMP-s) activity. Recently, it was proposed that some elastin derived hexapeptides resulting from circular permutations of VGVAPG (a three fold repetition sequence in exon 24 of human tropoelastin) possess MMP-1 production and activation regulation properties. This effect depends on the presence of the tropoelastin specific membraneous receptor 67 KDa EBP (Elastin Binding Protein). Our results obtained by using both circular dichroism spectroscopy and linear predictions confirmed the hypothesis of a structure dependent mechanism with a possibly occurring type VIII beta-turn on the first four residues of the GXXPG sequence consensus which is only present among all active peptides. Thus, we have performed extensive molecular dynamics studies, in both implicit and explicit solvent, on these active and inactive elastin derived hexapeptides. Using our own analysis method of pattern recognition of the types of the beta-reverse-turns followed during the molecular dynamics trajectory, we found that active and inactive peptides effectively form two well distinct conformational groups in which active peptides preferentially adopt conformation close to type VIII GXXP (beta-reverse-turn. The structural role of the C terminal G residue could also be explained. Additional molecular simulations on (VGVAPG)2 and (VGVAPG)3 show the formation of two or three GXXP tetrapeptides adopting a structure close to type VIII beta-reverse-turn, suggesting a local conformational preference for this motif. This observation of a specific structural single and/or repeated motif is in agreement with the circular dichroism spectra of the involved (VGVAPG)1, (VGVAPG)2 and (VGVAPG)3 peptides and then it can be proposed that their biological activities have to be linear. The final aim of this type of work is to understand more about the sequence/structure/function/activity relationships of those structured peptides in order to propose specific sequences (corresponding to specific structures) for best biological activity results. PMID- 11727708 TI - ECCO 11. The European Cancer Conference. Lisbon, Portugal, 21-25 October 2001. Abstracts. PMID- 11727709 TI - [Chemokines, tumor growth and angiogenesis]. AB - The immune surveillance represents an important defence mechanism of immediate destruction of the transformed malignant cells that daily arise in our body. Immune mechanisms play a great role also in the development of anti-tumour immunity. However, sometimes it occurs that the mechanisms of the immune system are misused by host cells for their own profit. This is also the case of chemokines. It was found out that malignant tumours produce chemokines with angiogenic activity which results in the formation of new blood vessels within the tumour. In this way the tumour ensures its blood supply. This not only enhances the tumour growth, however facilitates also the metastatic process. In addition to the latter the tumour growth is further enhanced by the influence of chemokines on the production of factors with immunosuppressive activity. (Fig. 3, Ref. 48.) PMID- 11727710 TI - [Pulmonary tuberculosis 1994-1998 at the Tuberculosis and Pulmonary Disease Clinic of Comenius University Medical School Hospital in Bratislava]. AB - The authors present the incidence of new cases of tuberculosis of the lungs treated at the Clinic of Tuberculosis and Lung Diseases in the years 1994-1998. This study evaluates the clinical forms of tuberculosis, rate of debacillisation, age stratification and the incidence of diseases associated with severe tuberculosis. PMID- 11727711 TI - [Methods of determining and evaluating reversibility of airway obstruction]. AB - The laboratory measurements of the reversibility of airways obstruction is assessed by performing pulmonary function tests before and after administration of bronchodilators. Controversial issues relating to bronchodilator testing include: selection of bronchodilator, dosage, pattern of inhalation of the drug, timing of the postbronchodilator test, physiologic tests utilized, criteria for the significant response, and the role of glucocorticoids in the relief of airways obstruction. PMID- 11727712 TI - [Latex allergy]. AB - Within the last ten years, the IgE mediated type of allergy to natural rubber latex has become an increasing and clinically important problem. Health professionals, rubber industry workers and individuals with spina bifida have been found to be at special risk of latex allergy. Clinical symptoms manifest as contact urticaria, rhinoconjunctivitis, asthma or even as intraoperative anaphylactic shock. An association between allergies to latex proteins and to various food-"Latex fruit syndrome "has been reported. Strict avoidance of the use of latex products is the primary preventive measure. In this review, the author pays attention to epidemiology, risk factors, clinical symptoms, diagnostics and prevention strategies. PMID- 11727713 TI - [The furosemide test in diagnosis of the cause of bronchial obstruction]. AB - The authors studied the rate of relief in cases of moderate, reversible airway obstruction after Furosemide administration. They examined 25 patients with bronchial asthma and chronic obstructive bronchitis (mean age 49 years). They found out that intravenous administration of Furosemide to patients with chronic obstructive bronchitis has a significant bronchodilating effect. The reduction of interstitial lung water and the decrease in congestion of airways mucosa in latent left ventricular failure may be the possible explanations of this phenomenon. After administration of Furosemide by inhalation, a mild decrease in airway obstruction was observed in asthmatic patients. The influence of Na+/K+/Cl transport in the bronchial epithelium, or changes in PGE2 are supposed to represent the effective mechanisms. The authors conclude that Furosemide given intravenously to patients with moderate airflow limitation in connection with lung function tests may be helpful in the diagnosis of the nature of airway obstruction. PMID- 11727714 TI - [Fournier's gangrene]. AB - Fournier's gangrene, although bearing the eponymous name of J.A. Fournier, was first reported by Baurienne in 1764 and 1883. His description was characterized by an abrupt onset of a rapidly progressive gangrene of the scrotum in otherwise healthy young males with an absence of a specific causative agent. Today, the frontiers of Fournier's gangrene have expanded to include all cases of necrotizing fasciitis of the perineum and external genitalia males and females from all age groups. All the patients were treated with antibiotics, surgical debridement and frequent wound dressings with hypertonis saline, hydrogen peroxide, colostomy and epicystostomy. PMID- 11727716 TI - Irish Endocrine Society 26th annual meeting. Killarney, Co. Kerry. 9-10 November 2001. Abstracts. PMID- 11727715 TI - [Personal experience with thyroid gland surgery and surgical complications]. AB - The authors from a department with numerous thyroid gland operations present their experience and operative complications. In this department, routine dissection of the recurrent laryngeal nerve is not performed, a method of a strietly subcapsular dissection of the gland with thorough preoperative haemostasis is used. Dissection of the nerve is performed only in necessary cases. Nevertheless, the operative complications do not differ from those of other departments with a similar field of interest. Emphasis is put especially on careful dissection and the surgeon's experience. PMID- 11727717 TI - Molecular targets and cancer therapeutics: discovery, biology, and clinical applications. October 29-November 2, 2001. Miami Beach, Florida, USA. Abstracts. PMID- 11727718 TI - Cumulative author and subject indexes 2000, volumes 852-887. PMID- 11727719 TI - [76th Annual meeting of the French Society of Surgical Orthopedics and Traumatology. Abstracts]. PMID- 11727720 TI - 8th Annual Conference of the International Society for Quality of Life Research. November 7-10, 2001. Amsterdam, The Netherlands. Abstracts. PMID- 11727721 TI - Principles of pharmaceutical management of spastic hypertonia. AB - Spasticity and hypertonicity are common components of the upper motor neuron syndrome (UMNS), which is seen in various neurologic conditions. Pharmacologic agents that can be administered through different routes can play an important part in treatment aimed at minimizing complications, pain, and functional deficits caused by UMNS. They can be administered by way of an oral, transdermal, intramuscular, or intrathecal route and can play an important role in the management of UMNS. This article reviews UMNS, the functional consequences that can result from it, individual agents, and their indications, complications, and adverse effects. Algorithms for treatment of UMNS with pharmacologic agents, and these agents' efficacy in different neurologic populations, is reviewed. PMID- 11727722 TI - Intrathecal baclofen therapy for spastic hypertonia. AB - Intrathecal baclofen is perhaps the most effective treatment for significant spasticity regardless of the origin. For appropriately selected patients, it can provide qualitative and quantitative improvements in quality of life. This article discusses the practical aspects and patient selection, trial, implant, and ongoing management of patients with intrathecal baclofen pump therapy. PMID- 11727723 TI - 19th World Congress on Endourology and SWL and 17th Basic Research Symposium. November 14-17, 2001. Bangkok, Thailand. Abstracts. PMID- 11727724 TI - International Society for Pharmacoeconomics and Outcomes Research (ISPOR)4th Annual European Conference. 11-13 November 2001. Cannes, France. Abstracts. PMID- 11727725 TI - [XXVII Meeting of the Polish Society of Pneumology. Abstracts]. PMID- 11727726 TI - A review of anticoagulation with warfarin in patients with nonvalvular atrial fibrillation. AB - BACKGROUND: Warfarin therapy has proved safe and effective in a number of randomized controlled trials of stroke prophylaxis in patients with nonvalvular atrial fibrillation (NVAF), reducing the risk of stroke in these patients by two thirds. However, participants in the clinical trials were carefully selected and younger than patients in actual clinical practice. OBJECTIVE: This analysis sought to determine whether the results of clinical trials in patients with NV can be extrapolated to the general population seen in clinical practice. METHODS: A MEDLINE search from 1966 to the present was used to identify observational trials of anticoagulation in patients with NVAF that addressed warfarin use, anticoagulation control, efficacy, and complications. The search terms used were atrial fibrillation and anticoagulation. RESULTS: Although warfarin prophylaxis against stroke in patients with NVAF appeared to be as well tolerated and effective in clinical practice as in clinical trials, it was generally underused, particularly in the elderly. Anticoagulation control was not as good in clinical practice as in clinical trials, although the rates of stroke and major bleeding were comparable. CONCLUSIONS: Judicious use of warfarin, tailored to individual stroke risk, seems to be a reasonable policy. Warfarin therapy increases quality adjusted survival in patients at high risk for stroke, and it is recommended for medium-risk patients unless their risk of bleeding is high or their quality of life while taking warfarin would be poor. Patients at a low risk for stroke will have equivalent health outcomes and incur lower costs if treated with aspirin. Despite the increased risk of hemorrhage in elderly patients, the net benefit of warfarin therapy is greater in this age group because of the higher risk of stroke. Active involvement of patients and their caregivers in an anticoagulation service setting may improve outcomes of anticoagulation therapy. PMID- 11727727 TI - Abstracts of the 8th Annual Meeting of the Oxygen Society. November 15-19, 2001. Research Triangle Park, North Carolina, USA. PMID- 11727728 TI - [Peutz-Jegher syndrome: the guilty speak]. PMID- 11727729 TI - [Again a dogma that collapses: that of the strictly cytoplasmic synthesis of proteins]. PMID- 11727730 TI - ECTRIMS 2001. 17th Congress of the European Committee for Treatment and Research in Multiple Sclerosis. 12-15 September 2001. Dublin, Ireland. Abstracts. PMID- 11727731 TI - American Heart Association 2001 scientific sessions: late-breaking science-drug eluting stents. PMID- 11727732 TI - American Heart Association 2001 scientific sessions: late-breaking science-ACE inhibitor therapy in African-Americans. PMID- 11727733 TI - 7th Nottingham International Breast Cancer Conference. 18-21 September 2001. Abstracts. PMID- 11727734 TI - Clinical protocol. Purging of autologous stem cell sources with bcl-x(s) adenovirus for women undergoing high-dose chemotherapy for stage IV breast carcinoma. AB - High-dose chemotherapy (HDCT) and autologous bone marrow transplantation (BMT) is frequently used to treat patients with metastatic cancer including breast cancer and neuroblastoma. However, the bone marrow of such patients is often contaminated with tumor cells. Recently, we have found that a recombinant adenovirus vector that contains a bcl-x, minigene (a dominant negative inhibitor of the bcl-2 family), called the bcl-x(s) adenovirus, is lethal to cancer cells derived from epithelial tissues, but not to normal human hematopoietic cells. To determine the mechanism, by which this virus spares normal hematopoietic cells, we isolated normal mouse hematopoietic stem cells and infected them with an adenovirus that contains a beta-galactosidase minigene. Such cells do not express beta-galactosidase, indicating that hematopoietic stem cells do not express transgene encoded by adenovirus vectors based upon the RSV-AD5 vector system. When breast cancer cells mixed with hematopoietic cells were infected with the bcl-x(s) adenovirus, cancer cells were selectively killed by the suicide adenoviruses. Hematopoietic cells exposed to the suicide vectors were able to reconstitute the bone marrow of mice exposed to lethal doses of y-irradiation. These studies suggest that adenovirus suicide vectors may provide a simple and effective method to selectively eliminate cancer cells derived from epithelial tissue that contaminate bone marrow to be used for autologous BMT. We therefore propose to initiate a phase I clinical trial to test the safety of this virus in women with breast cancer undergoing high does chemotherapy and autologous BMT. PMID- 11727735 TI - Clinical protocol. A phase IIb, randomized, multicenter, double-blind study of the efficacy and safety of Trinam (EG004) in stenosis prevention at the graft vein anastomosis site in dialysis patients. AB - Hemodialysis access complications remain a major cause of morbidity for patients with end-stage renal disease who are undergoing chronic hemodialysis. Vascular access complications occur in approximately 40% of patients with polytetrafluorethylene (PTFE) grafts within the first 6 months, primarily due to stenosis and thrombosis. Thrombosis at the site of vascular access increases the risk of infection and the need for hospitalization, and may lead to loss of potential new sites for vascular access. To a large extent, the failure of hemodialysis access is due to the rapid development of an intimal hyperplastic lesion in the region of anastomosis between the PTFE graft and the vein. The hospital costs related to hemodialysis access procedures are estimated to be around $1.3 billion per year and the total cost of hemodialysis complications to the US healthcare system is thought to be in excess of $2 billion per year. Ark Therapeutics Ltd. are developing a vascular endothelial growth factor D (VEGF-D) gene in an adenoviral vector which is delivered locally to the adventitial surface of a graft-vein anastomosis by means of a collagen collar device. The proposed indication for this product (Trinam) is the prevention of intimal hyperplasia at the graft-vein anastomosis site in patients who require vascular access to facilitate hemodialysis for end-stage renal disease. The rationale for Trinam to prevent intimal hyperplasia at the graft-vein anastomosis follows the discovery that VEGF has a 'vasculoprotective' action, resulting in inhibition of smooth muscle cell migration and proliferation. The fundamental mechanism for this vasculoprotective effect of VEGF, as distinct from its more widely appreciated 'angiogenic' role, is that VEGF acts on surface receptors on endothelial cells resulting in increased production of nitric oxide and prostacyclin. These entities diffuse into the media of the blood vessel wall and counter the tendency for intimal hyperplasia to develop. In an in vivo rabbit model of intimal thickening in carotid arteries, adventitial delivery of VEGF using a silastic collar as a gene delivery reservoir prevented smooth muscle cell proliferation without evidence of new blood vessel formation, indicating that the mechanism by which VEGF inhibited intimal hyperplasia did not involve angiogenesis. The objective of the proposed study is to assess the efficacy and safety of local delivery of Trinam when applied to the graft-vein anastomosis site in patients with end-stage renal disease who require vascular access for hemodialysis. At the time of surgical placement of a PTFE arm graft, patients will be randomized to either a single administration of Trinam or to 'no treatment' (i.e., control group). It is hypothesised that Trinam administration will result in less stenosis at the graft-vein anastomosis site (as measured by fistulography) compared with controls and therefore will reduce the need for interventions in dialysis patients. Approximately 210 patients will be enrolled from 10-15 centers and patients will be evaluated for efficacy and safety over 6 months. The total dose of Trinam will be 1 x 10(11) viral particles (replication deficient adenoviral vector). This dose of Trinam was not associated with any significant toxicology findings in a preclinical study of pigs in which a PTFE loop-graft was anastomosed from the carotid artery to the internal jugular vein to mimic hemodialysis vascular access surgery. PMID- 11727736 TI - Clinical protocol. A phase 1 open-label clinical trial of the safety and tolerability of single escalating doses of autologous CD4 T cells transduced with VRX496 in HIV-positive subjects. AB - The proposed study is a Phase I trial to evaluate a HIV-based lentiviral vector carrying an antisense sequence targeted to HIV in the treatment of HIV infection. The primary objective of this Phase I study is to determine the safety and tolerability of treatment with autologous CD4+ T cells modified (transduced) ex vivo with VRX496 when administered to HIV-infected patients. VRX496 is a completely gutted lentiviral vector and does not code for any viral proteins. The viral vector contains an antisense sequence targeted to the HIV envelope (env) gene. VRX496 directly interferes with wild-type HIV (wt-HIV) expression via anti env antisense expression in vector transduced CD4 cells that become infected with wt-HIV. Expression of the anti-HIV antisense env from a HIV vector transcript would target wt-HIV RNA and destroy it, and hence, decrease productive HIV replication from CD4 T cells. The clinical goal for this treatment approach is to decrease viral loads and promote CD4 T cell survival in vivo. Data from in vitro studies suggest that HIV vectors such as VRX496 could potentially reduce viral loads in HIV-infected individuals and thus could delay the onset to AIDS while promoting CD4 T cell survival and providing the immune system with a better chance to control the infection. Additionally, preliminary results from experiments in SCID mice (mice with transplanted human immune cells) indicate that the human cells transduced with VRX496 and implanted into the SCID mice do not elicit any overt adverse effects. HIV-infected patients (CD4 T cell count of >200/mm3, discontinued from HAART therapy) will undergo leukoapheresis with subsequent CD4 T cell isolation. Patient CD4 T cells will be transduced ex vivo with the vector, expanded for 8-11 days, and then the modified cells will be reintroduced into the patient. Each subject will receive a single intravenous injection infused over 30 minutes; subjects will be examined 24, 48, and 72 hours post-injection and weekly for 4 weeks. Patients will receive one of four different ascending doses (1 x 10(9), 3 x 10(9), 1 x 10(10), and 3 X 10(10) cells/patient). Doses will be administered to four independent, sequential subject cohorts of 3 patients. Groups will be administered escalating doses at 6 week intervals after safety has been demonstrated in the previous group. Follow up examinations will be conducted 1, 3, and 6 months post-injection. Long term follow-up including RCR testing will be performed. PMID- 11727737 TI - Clinical protocol. An open-label, phase I, single administration, dose-escalation study of ADGVPEDF.11D (ADPEDF) in neovascular age-related macular degeneration (AMD). AB - Age-Related Macular Degeneration (AMD) is, together with Diabetic Retinopathy, the most common cause of vision loss among adults in the U.S. and other developed countries. In the U.S., at least 1.7 million people have impaired vision due to AMD. Every year, more than 165,000 people contract AMD and 16,000 go blind from it, predominantly from a rapidly progressing form of the disease called "wet" AMD. Wet AMD is characterized by serious or hemorrhagic detachment of the retinal pigment epithelium and choroidal neovascularization. The macula has the highest concentration of photoreceptors facilitating central vision and permitting high resolution visual acuity. The damage caused by the leakage and fibrovascular scarring in wet AMD leads to profound loss of central vision and severe loss of visual acuity (usually 20/200 or worse). People with wet AMD have several limitations, including inability to read, inability to recognize faces or drive, and the disease often leads to blindness. The onset of severe visual changes in wet AMD can occur suddenly. More than 400,000 Americans are currently affected by this form of the disease, and the incidence is rising rapidly with the aging of the population. Therefore, the serious consequences of this disease along with the limited treatment options and their effectiveness make this a very good candidate for a gene transfer treatment approach. The investigational agent, Ad(GV)PEDF.11D, is an E1-, partial E3-, E4- deleted replication-deficient, adenovirus serotype 5, gene transfer vector. The transgene in this vector is the cDNA for human pigment epithelium-derived factor (PEDF). PEDF is one of the most potent known antiangiogenic proteins found in humans. While Ad(GV)PEDF.11D is able to transduce many somatic cell types, the natural barrier to other tissues created by the retina limits the ability of Ad(GV)PEDF.11D to affect tissues other than in the eye. Intravitreal administration of Ad(GV)PEDF.11D provides a convenient means of delivering PEDF to the relevant cells within the eye likely to result in a more prolonged duration of effect versus administration of the PEDF protein alone. In three murine disease models (the laser-induced choroidal neovascularization model, the VEGF transgenic model, and the retinopathy of prematurity model) significant inhibition of neovascularization (up to 85%) was demonstrated with doses of Ad(GV)PEDF vectors ranging from 1 x 10(8) to 1 x 10(9) pu. In toxicology studies performed in Cynomolgus monkeys, a dose-related inflammatory response was observed. A dose of 1 x 10(8) pu caused no adverse effects, while the inflammatory response observed at 1 x 10(9) pu was minimal and fully reversible. The observed inflammatory response at doses of 1 x 10(10) and 5 x 10(10) pu were increasingly severe. The proposed clinical trial is an open label, dose-escalation, phase I study to investigate the safety, tolerability and potential activity of intravitreal injection of Ad(GV)PEDF.11D in patients with wet AMD. Ad(GV)PEDF.11D will be injected once via intravitreal injection into the eye with the most advanced AMD based on visual acuity. Subjects will be age 50 or over and have severe wet AMD in at least one eye defined by a best-corrected vision of 20/200 or worse. The primary objectives of this investigation are: (1) to assess the safety, tolerability and feasibility of intravitreal injection of Ad(GV)PEDF.11D in patients with severe, neovascular AMD, (2) to identify the maximum tolerated dose (MTD) of Ad(GV)PEDF.11D, and (3) to get some indication of potential activity of Ad(GV)PEDF.11D. PMID- 11727738 TI - American Society of Hematology 43rd Annual Meeting. December 7-11, 2001. Orlando, Florida, USA. Abstracts. PMID- 11727740 TI - 142nd Meeting of the Acoustical Society of America. Fort Lauderdale, Florida, USA. 3-7 December 2001. Abstracts. PMID- 11727739 TI - American Society of Hematology 43rd Annual Meeting. December 7-11, 2001. Orlando, Florida, USA. Abstracts. PMID- 11727741 TI - 41st American Society for Cell Biology Annual Meeting. December 8-12, 2001. Washington, DC, USA. Abstracts. PMID- 11727742 TI - Psycho Neuro Endocrino Immunology Meeting. November 15-17, 2001. Regensburg, Germany. Abstracts. PMID- 11727744 TI - Abstracts of original communications from the Nutrition Society meetings. March, June, July 2001. PMID- 11727743 TI - 34th Annual Undersea and Hyperbaric Medical Society Scientific Meeting. 14-16 June 2001. Abstracts. PMID- 11727745 TI - [6th Francophile meeting on Alzheimer disease and the international meeting on dementia of the French Society of Neurology. Bordeaux, France, 9-13 October 2001. Abstracts]. PMID- 11727746 TI - 1st Joint French-German Congress on Cell Biology. 7-9 November 2001, Strasbourg, France. Abstracts. PMID- 11727747 TI - Cumulative index, volumes 1-99. PMID- 11727748 TI - Medicare program; prospective payment system and consolidated billing for skilled nursing facilities--update. Final rule. AB - This final rule updates the payment rates used under the prospective payment system (PPS) for skilled nursing facilities (SNFs) for fiscal year (FY) 2002, as required by statute. Annual updates to the PPS rates are required by section 1888(e) of the Social Security Act (the Act), as amended by the Medicare, Medicaid, and SCHIP Balanced Budget Refinement Act of 1999 (BBRA), and the Medicare, Medicaid, and SCHIP Benefits Improvement and Protection Act of 2000 (BIPA), relating to Medicare payments and consolidated billing for SNFs. As part of this annual update, we are rebasing and revising the routine SNF market basket to reflect 1997 total cost data (the latest available complete data on the structure of SNF costs), and modifying certain variables for some of the cost categories. Finally, we are implementing the transition of swing-bed facilities to the SNF PPS, effective with cost reporting periods beginning on and after July 1, 2002. PMID- 11727751 TI - Are you in the right plan? More than 83,000 readers tell us about their experiences in managed care. PMID- 11727750 TI - Rotation and direction judgment from visual images head-slaved in two and three degrees-of-freedom. AB - The contribution to spatial awareness of adding a roll degree-of-freedom (DOF) to telepresence camera platform yaw and pitch was examined in an experiment where subjects judged direction and rotation of stationary target markers in a remote scene. Subjects viewed the scene via head-slaved camera images in a head-mounted display. Elimination of the roll DOF affected rotation judgment, but only at extreme yaw and pitch combinations, and did not affect azimuth and elevation judgement. Systematic azimuth overshoot occurred regardless of roll condition. Observed rotation misjudgments are explained by kinematic models for eye-head direction of gaze. PMID- 11727753 TI - Adult vaccines: missed shots. PMID- 11727754 TI - Targeting the brain's immune system: a psychopharmacological approach to central nervous system infections. PMID- 11727755 TI - Mitogen-induced activation of mouse T cells increases kappa opioid receptor expression. PMID- 11727756 TI - Self-enhancement of phagocytosis by murine resident peritoneal macrophages and its relationship to morphine effects on the process. PMID- 11727757 TI - Role of mu-opioid receptor in immune function. PMID- 11727758 TI - Opiates promote T cell apoptosis through JNK and caspase pathway. AB - Opiate addicts are prone to recurrent infections. In the present study we evaluated the molecular mechanism of opiate-induced T cell apoptosis. Both morphine and DAGO ([D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin) enhanced T cell apoptosis. Morphine as well as DAGO activated c-Jun NH2-terminal kinase (JNK) in T cells. Moreover, opiates increased the expression of ATF-2. a specific substrate for JNK and P38 mitogen activated kinases (MAPK). Furthermore, opiates attenuated extracellular signal related kinase (ERK) in T cells. Both morphine and DAGO cleaved pro-caspases 8, 9, and 10 and generated caspases 8, 9 and 10 (active products). Morphine as well as DAGO also cleaved poly-(ADP-ribose) polymerase (PARP) into 116 and 85 kD proteins indicating the activation of caspase-3. These results suggest that opiate-induced T cell apoptosis may be mediated through the JNK cascade and activation of caspases 8 and 3. PMID- 11727759 TI - Role of beta-endorphin in the modulation of immune responses: perspectives in autoimmune diseases. PMID- 11727760 TI - Modulation of fas/fasL in a murine retroviral infection by AZT and methionine enkephalin. PMID- 11727761 TI - Acute effects of heroin on the cellularity of the spleen and the apoptosis of splenic leukocytes. PMID- 11727762 TI - Alteration of early T cell development by opioid and superantigen stimulation. PMID- 11727763 TI - Effect of opioids on oral Salmonella infection and immune function. PMID- 11727764 TI - Altered T-cell responsiveness in morphine "tolerant" rats: evidence for a potential role of the paraventricular nucleus of the hypothalamus. PMID- 11727765 TI - Actions of endotoxin and morphine. AB - In summary, our current studies show that treatment with a bacterial endotoxin, lipopolysaccharide (LPS), induces the expression of mu opioid receptors in the rat mesentery. This induction may be mediated through IL-1's actions on mu opioid receptors. Morphine stimulates the expression of adhesion molecules in human brain microvascular endothelial cells (HBMEC) isolated from pathological tissues. Under pathological conditions, mu opioid receptor-dependent pathways may be modulated through the induction of mu opioid receptors, especially in endothelial cells. Treatment with morphine increases [14C]-inulin permeability of an in vitro microvascular endothelial cell barrier, and decreases endothelial cell viability. Morphine pre-treatment potentiates the effects of LPS on endothelial cell viability, and on LPS induction of IL-1beta secretion from 1alpha, 25-dihydroxy vitamin D3-treated HL-60 human leukemia cells. Previously, it was suggested that an opioid-dependent pathway may be involved in the recovery from endotoxin shock (D'Amato and Holaday, 1984). Induction of mu opioid receptors by treatment with high doses of endotoxin suggests that mu opioid receptor-dependent pathways may be involved in mediating the response to endotoxins. Taken together, these data provide valid evidence for an association between endotoxins and opioid actions. These studies suggest that opioid-dependent pathways in disease or in endotoxin exposure may be modified by cytokine-induced expression of opioid receptors in endothelial cells. In a pathological condition, an alteration of the opioid dependent pathway may be expected. When morphine is used for its therapeutic values, it may, indeed, potentiate LPS' effects in an adverse manner. From a clinical perspective, these data indicate that morphine and an endotoxin, such as LPS, may interact in a positive 'feedback type of reaction, and thereby modulate the body's immune responses with unexpected and detrimental results. PMID- 11727766 TI - Pharmaconeuroimmunology in the intestinal tract: opioid and cannabinoid receptors, enteric neurons and mucosal defense. PMID- 11727767 TI - Cannabinoid-mediated inhibition of inducible nitric oxide production by rat microglial cells: evidence for CB1 receptor participation. AB - Activated brain microglial cells release inflammatory mediators such as nitric oxide (NO) that may play important roles in central nervous system antibacterial, antiviral, and antitumor activities. However, excessive release of these factors has been postulated to elicit immune-mediated neurodegenerative inflammatory processes and to cause brain injury. Recent studies using the rat animal model indicate that select cannabinoids may modulate production of these inflammatory factors. Treatment of neonatal rat brain cortical microglial cells with the cannabinoid paired enantiomers CP55940 and CP56667 resulted in a stereoselective differential effect on inducible NO production. The analog CP55940 exerted a dose dependent inhibition of interferon gamma (IFNy)/bacterial lipopolysaccharide (LPS)-inducible NO production which was significantly greater than that exerted by CP56667. Pretreatment of microglial cells with the CB1 cannabinoid receptor selective antagonist SR141716A reversed this CP55940-mediated inhibition. MRT-PCR demonstrated the presence of CB1 receptor mRNA within microglial cells consistent with the presence of CB1 receptors. Collectively, these results indicate that the cannabinoid analog CP55940 selectively inhibits inducible NO production by microglial cells and that this inhibition is effected, at least in part, through the CB1 receptor. PMID- 11727768 TI - Modulation of CB1 mRNA upon activation of murine splenocytes. AB - There is significant evidence that cannabinoids have the ability to exert immunomodulatory effects. The identification of cannabinoid receptors in immune tissues has therefore led to questions about whether these immunomodulatory effects occur via these cannabinoid receptors. The cannabinoid receptor 1 (CB 1), although expressed primarily in the brain, is also expressed in lower amounts in peripheral tissues. Of interest to us is the fact that CB1 is expressed in immune tissues such as spleen, albeit at lower levels than the peripheral cannabinoid receptor, CB2. To examine the function of CBI in immune cells, activation experiments were performed using different stimuli e.g., anti-CD3, phorbol 12 myristate 13-acetate (PMA)/Ionomycin (Io), and PMA/Io + IL-2. Whole spleen cells were cultured in the presence of different stimuli for 0, 2, 4, and 24 hours, harvested at each time point, RNA isolated, and RT-PCR performed. FACS analysis was also performed using CD69 (an early activation marker) to determine whether cells were actually being activated. Results from anti-CD3 stimulation indicated a decrease in CB1 mRNA expression following activation. CB1 mRNA expression in murine splenocytes that were stimulated with PMA/Io in the presence or absence of IL-2 was also modulated. Expression of the message was enhanced upon stimulation with PMA/Io and PMA/Io + IL-2, however, stimulation with PMA/Io + IL-2 led to a stronger increase within 2 to 4 hours with CB1 returning to at or below baseline levels by 24 hours. Expression of CD69 was detected in all stimulated samples thereby indicating that the splenocytes were becoming activated. In summary, anti CD3 stimulation appeared to decrease CB1 mRNA expression while PMA/Io + IL-2 stimulation significantly increased CB1 mRNA expression. These results demonstrate that the expression of CB1 mRNA is modulated upon cellular activation and that this modulation is dependent on the stimulus that is used. PMID- 11727769 TI - Downregulation of cannabinoid receptor 2 (CB2) messenger RNA expression during in vitro stimulation of murine splenocytes with lipopolysaccharide. AB - Cannabinoid receptor 2 (CB2) has been identified as the most abundant cannabinoid receptor subtype in the immune system. Bacterial lipopolysaccharide (LPS) is a potent stimulant of B cells, inducing proliferation and differentiation into antibody secreting cells. It has been reported that CB2 receptor expression is upregulated during human, tonsillar B cell activation through CD40. It was of interest to investigate the expression of CB2 mRNA using another B cell activator, LPS. Using northern blot analysis, we measured CB2 mRNA levels in murine splenocytes and enriched B cells. Results indicated that the 4.0 kb CB2 transcript was 2 fold higher in abundance in murine B cells than in whole splenocyte preparations. This observation confirmed data from others and from our previous RT-PCR studies that the expression of CB2 mRNA is more abundant in B cells. Upon LPS stimulation, CB2 transcripts were decreased 46% and 42% at 4 hours and 24 hours, respectively, when compared to unstimulated populations. An examination by flow cytometry of the CD69, early activation marker, on splenocytes, showed that the majority of the B cells were activated at 24 hrs. Thus, these results suggested that LPS stimulation of murine B cells caused a decrease in CB2 mRNA expression in contrast to the increase observed following human B cell stimulation through CD40. PMID- 11727770 TI - CB1 and CB2 receptor mRNA expression in human peripheral blood mononuclear cells (PBMC) from various donor types. AB - Marijuana cannabinoid receptors (CBR), CB1 and CB2, are G protein-coupled receptors reported to be expressed in brain as well as cells of the periphery. Human peripheral blood mononuclear cells (PBMCs) are reported to express CBR mRNA with CB2 expression higher than CB1 and expression in B cells higher than other cells. However, it is not known if the mRNA expression is constant among individuals of differing ages, gender, or ethnic origins. In the present study, PBMCs were obtained from a limited number of normal donors of both genders, of ages ranging from 21 to 55, and from Caucasian, and Asian ethnic origin. Using semi-quantitative RT-PCR, we confirmed previous reports that CB2 mRNA expression was higher than CB1 in PBMCs and in addition demonstrated that this basic profile was observed when stratified by age, gender, or ethnic origin. The latter results suggest that CBR expression is relatively constant across the human population. PMID- 11727771 TI - Effect of cocaine on chemokine and CCR-5 gene expression by mononuclear cells from normal donors and HIV-1 infected patients. PMID- 11727772 TI - Intravenous cocaine abuse: a rodent model for potential interactions with HIV proteins. PMID- 11727773 TI - Substance P receptor mediated macrophage responses. AB - Taken together, these studies demonstrate an important role for substance P receptor expression by macrophages. The results to date suggest proinflammatory signals mediated by this receptor, and it is clear that substance P can act synergistically with other factors to stimulate macrophage activity. Antagonism of substance P/substance P receptor interactions in vivo profoundly affect immunity against Salmonella. This model provides evidence that an optimal host response against this intracellular pathogen of macrophages requires signaling through the substance P receptor. The ability of interferon gamma or IL-4 to upregulate substance P receptor mRNA expression on macrophages suggests that substance P-mediated amplification loops might involve either T helper type 1 or T helper type 2 responses. Thus, depending upon the immunologic stimulus, substance P could contribute to cell mediated as well as humoral immune responses. Several important questions remain. Since the antigen processing and presenting function is an important macrophage activity, the effect of signaling through the substance P receptor on these events has not been defined. Furthermore, since macrophages are only one type of antigen presenting cell, it will be important to determine the role of substance P receptor expression in the activity of dendritic cells. We anticipate that these ongoing investigations will further define the positive contributions that substance P/substance P receptor interactions have in the initiation of immune responses. PMID- 11727775 TI - NeuroAIDS: retroviral pathology and drugs of abuse. PMID- 11727774 TI - Lymphocyte modulation by seven transmembrane receptors: a brief review of session 1. PMID- 11727776 TI - The effects of drugs and neuropeptides on immunomodulation and opportunistic infection: summary of symposium 2. PMID- 11727777 TI - Direct and indirect mechanisms of HIV-1 neuropathogenesis in the human central nervous system. PMID- 11727778 TI - Chemokine receptors on brain endothelia--keys to HIV-1 neuroinvasion? PMID- 11727779 TI - Neurotrophic factor regulation of human immunodeficiency virus type 1 replication in human blood-derived macrophages through modulation of coreceptor expression. PMID- 11727780 TI - Direct vs. indirect modulation of complex in vitro human retroviral infections by morphine. PMID- 11727781 TI - Genetic factors involved in central nervous system/immune interactions. AB - Analysis of several inbred rat strains has led us to hypothesize that HPA axis abnormalities may contribute, in part, to susceptibility to both autoimmune disease and addiction. In this article we review the evidence for this hypothesis and describe our ongoing efforts to genetically characterize these traits. We have mapped the locations of 23 loci that regulate autoimmune disease in rats, and are currently constructing QTL congenic lines in which a genomic region from the resistant strain is transferred to the susceptible strain or vice versa. These QTL congenic lines will be valuable to test whether genes encoding autoimmune regulation also control neuroendocrine traits. Further genetic dissection and identification of the underlying genes will be necessary to infer a mechanistic link between autoimmune and neuroendocrine traits. PMID- 11727782 TI - Interactions of opioid receptors, chemokines, and chemokine receptors. PMID- 11727784 TI - Crosstalk between chemokine and opioid receptors results in downmodulation of cell migration. PMID- 11727783 TI - Model systems for assessing cognitive function: implications for HIV-1 infection and drugs of abuse. AB - Memory deficits are common among drug abusers and in those with chronic neurodegenerative disorders. Currently, the mechanisms through which diverse neurophysiologic processes alter memory are not known. This review describes the current systems and rationale for studying memory formation, consolidation, and recall. Special attention is given to physiologic (hippocampal long-term potentiation) and behavioral animal models. The principles and methods described can be applied to studies of diverse clinical disorders. PMID- 11727785 TI - Morphine upregulates kappa-opioid receptors of human lymphocytes. AB - Opioids such as morphine are potent analgesic and addictive compounds. Chronic morphine use also induces immunomodulatory and immunosuppressive effects, as especially evident in HIV-infected patients. Morphine acts on the immune cells primarily through its binding to mu-opioid receptors on the plasma membrane. However, morphine modulation of immune functions still exists in mu-opioid receptor knockout mice, suggesting that in addition to the mu opioid receptors, morphine may also act by mechanisms mediated by either delta or kappa opioid receptors. To determine whether morphine activates kappa opioid receptors (KOR), a quantitative competitive RT-PCR procedure was utilized to quantify the KOR gene expression of morphine-treated cells. A segment of KOR transcript spanning the second extracellular loop, which has the reported dynorphin specificity, and the seventh transmembrane domain of the receptor was amplified from the total RNA of morphine-treated CEM x174 lymphocytes, along with a competitor molecule. The competitor was constructed by deleting a 33-nucleotide fragment from KOR. The results of the competitive RT/PCR indicated that CEM x174 cells expressed KOR mRNA constitutively, in the order of femto-grams. Treatment of 10 microM of morphine resulted in the up-regulation of KOR gene expression 24 hr post treatment. The observed morphine effect could be reversed by treating the cells with either naloxone (a KOR-partially selective antagonist) or nor Binaltorphimine (a KOR-selective antagonist). PMID- 11727786 TI - Effects of morphine on T-cell recirculation in rhesus monkeys. AB - A 2-yr study on effects of morphine on lymphocyte circulation in rhesus monkeys (Macaca mulatta) showed that, over time, a well-maintained morphine-dependency caused biphasic depressive effects on circulating lymphocyte levels. Depression of T cell circulation by opiates actually was a relative effect. Morphine exposure basically stabilized T cell circulation in the context of concurrent increases in controls. Biphasic effects of morphine were attributable to distinctions in circulation kinetics of CD4+/CD62L (+ & -) T cells. That is, levels of CD4+/CD62L+ T cells were selectively depressed by opiates through the first 32wk after initiation of drug, and levels of CD4+/CD62L- T cells were selectively depressed thereafter. Regression analyses also showed that morphine stabilized lymphocyte recirculation. Circulating levels of resting and activated memory types of T cells were positively correlated in opiate-exposed monkeys during the first 32wk after opiate exposure--an effect not seen with control monkeys. Considerations of changes in the types of experimental stressors extant during the study suggested that temporally differential effects of opiates on T cell recirculation were connected with changes in the stress environment and the ability of morphine to modulate these changes. Thus, morphine, and by inference the endogenous opioid system, are involved in homeostasis of lymphocyte recirculation, probably through effects on central mediation of the stress axis. PMID- 11727787 TI - Chlorzoxazone: a probe drug the metabolism of which can be used to monitor one point blood sampling in the carbon tetrachloride-intoxicated rat. AB - In this study, we have carried out an investigation to determine if chlorzoxazone (CZX) is a suitable probe drug for predicting hepatic injury in carbon tetrachloride (CCl4)-intoxicated rats. The animals received oral doses of CCl4 (0.25, 0.5 and 1 ml/kg) 24 h prior to intraperitoneal administration of CZX. The total CYP and CYP2E1 content, as well as the aniline and CZX hydroxylase activity (Vmax and CLint), was reduced depending on the dose of CCl4 administered. At the highest concentration (128 mM) of diethyldithiocarbamate, a specific inhibitor of CYP2E1, the production of 6-hydroxychlorzoxazone (HCZX) in microsomes from CCl4 treated rats was reduced by about 85%. The IC50 value in microsomes from CCl4 treated rats was between 3 and 5 microM. The production of HCZX and the activity of aniline hydroxylase in CCl4-treated rats correlated with the amount of rat CYP2E1 protein (r=0.881, P<0.001 and r=0.822, P<0.001, respectively). The elimination of CZX by CCl4-treated rats was reduced and the HCXZ production in the CCl4-treated group was less than that in the olive oil-treated control group. The correlations between the intrinsic clearance [CLint: Vmax/Km) in vitro and the total body clearance (CLtot) of CZX hydroxylation and the elimination half life (t1/2) of CZX in vivo in CCl4-treated rats were high (r=0.839, P<0.001 and r= -0.828, P<0.001, respectively). In addition, the metabolic plasma HCZX/CZX ratio did not require multiple blood sampling and, 2 h after CZX administration in vivo, there was also a high correlation with the CLint (Vmax/Km) in vitro (r= 0.909, P<0.01). In conclusion, these results from this study demonstrate that CZX is a good probe for monitoring the inhibition of metabolism in rats due to CCl4 treatment. PMID- 11727788 TI - Repeated administration of a mild acute toxic dose of di-n-butyltin dichloride at intervals of 3 weeks induces severe lesions in pancreas and liver of rats. AB - Di-n-butyltin dichloride (DBTC) induced thymus atrophy, bile duct lesions, pancreatitis, and liver lesions in rats. Depending on dose [6 and 8 mg/kg intravenous (i.v.) DBTC] and time (1-24 weeks), the lesions in pancreas developed to a pancreatic fibrosis and the lesions in liver to liver cirrhosis. A single i.v. administration of 4 mg/kg DBTC induces a mild interstitial pancreatitis after 2-4 days followed by a restitutio ad integrum after 21-28 days. In the present study, the lesions of biliopancreatic duct, pancreas, and liver of rats after repeated administration of 4 mg/kg DBTC i.v. at intervals of 3 weeks have been investigated. The histopathological changes of pancreas and liver were examined by light microscopy 1,4, and 7 days and 2,3,4,6,9, and 12 weeks after administration of DBTC. Furthermore, pathobiochemical parameters of pancreatitis (amylase and lipase activity in serum), liver lesions (alkaline phosphatase activity and bilirubin in serum), and of fibrosis (hyaluronic acid in serum) were studied. Repeated administration of rats with DBTC (4 mg/kg i.v.) at intervals of 3 weeks induced an acute interstitial pancreatitis and after 9-12 weeks, a pancreatic fibrosis and liver lesions (intrahepatic bile duct hyperplasia, inflammation in periportal tract, and necrosis). In serum, elevated levels of alkaline phosphatase, bilirubin, and hyaluronic acid were found. This study demonstrates that the organotin compound induces toxic effects on pancreas and liver of rats by repeated administration of lower doses at long intervals. The risk of exposure to organotin at long intervals should be considered. PMID- 11727789 TI - Reproductive toxicity of DDT in adult male rats. AB - The reproductive toxicity of DDT was investigated in adult male rats exposed to 50 and 100 mg/kg body weight (b.wt) day(-1) for 10 successive days. Compared with control animals, administration of DDT led to a dose-dependent reduction of testicular weight and the number as well as the percentage of motile spermatozoa in the epididymis. Testicular histological observations revealed also a marked loss of gametes in the lumen of seminiferous tubules. In DDT-treated rats, the seminal vesicles weights dropped significantly, resulting from a decrease of testosterone production by testes, whereas serum LH and FSH increased after pesticide exposure. This increase of gonadotrophin levels may be related to an impairment of the negative feedback exerted by the steroid on the hypothalamic- pituitary axis. It is concluded that DDT induced adverse effects on male rat fertility by acting directly on the testes and altering the neuroendocrine function. PMID- 11727790 TI - Lipid peroxidation as a possible secondary mechanism of sterigmatocystin toxicity. AB - Sterigmatocystin (Stg), a major secondary metabolite of Aspergillus versicolor and A. nidulans, is the precursor of aflatoxin B1. In this study, male albino rats were treated with Stg-contaminated diet for 30 days, resulting in reduced levels of glutathione, ascorbic acid, and alpha-tocopherol. The activity of catalase in liver was reduced, whereas an increase in the activities of superoxide dismutase and glutathione peroxidase was observed. The levels of cytochrome P450, cytochrome b5, cytochrome b5 reductase, cytochrome c reductase, hydroxyl radical, and hydrogen peroxide formation significantly increased in the Stg- treated rat liver microsomes. Hepatic parenchymal cell injury, necrosis, and Kupffer cells proliferation were noticed in histological sections of liver from animals treated with Stg. Overall results suggest that generation of free radicals imposes depletion of antioxidants. This led to enhanced lipid peroxidation. The observed elevation of hepatic thiobarbituric acid reactive substances appears to originate mainly from the damaged Kupffer cells. As a result, elevated levels of serum marker enzymes were also observed. PMID- 11727791 TI - Factors affecting R6 fungicide toxicity on sea urchin fertilization and early development: roles of exposure routes and mixture components. AB - A technical fungicide mixture, R6 and its components, cymoxanil (CYM) and cupric oxychloride (Cu-OCl), were tested by sea urchin bioassays (Paracentrotus lividus and Sphaerechinus granularis). A set of toxicity endpoints was evaluated including both lethal and sublethal effects with the following endpoints: (a) acute embryotoxicity, (b) developmental defects, (c) changes in sperm fertilization success, (d) transmissible damage from sperm to the offspring, and (e) cytogenetic abnormalities. Acute effects on developing embryos were observed as early (prehatch) mortality at R6 levels > or =25 microg/ml. The pesticide mixture R6 was tested at realistic concentrations, ranging from 25 ng/ ml to 2.5 microg/ml, and the two components, CYM and Cu-OCl, were tested, either alone or in mixture, at concentrations equal to their levels in the corresponding R6 solutions. R6 was either dissolved in filtered seawater (water only, W-O), or spiked in "pristine" silt-clay sediment or soil samples before bioassays. Developmental toxicity of R6, following W-O dissolution, displayed a significant dose-related increase of larval malformations and differentiation arrest at concentrations of 750 ng/ml to 2.5 microg/ml both in P. lividus and in S. granularis larvae. Developmental toxicity was removed in spiked sediment up to R6 nominal levels (25 microg/ml), 10-fold above the embryotoxic R6 levels in W-O exposure. No significant developmental toxicity was exerted by CYM or Cu-OCl (W-O exposure) up to their concentrations equivalent to 2.5 microg/ml R6. The laboratory-prepared mixture of CYM and Cu-OCl, in the same concentration range, only resulted in minor effects, as larval retardation, suggesting the presence of toxic impurities (or additional components) in the R6 formulation. When sperm from either P. lividus or S. granularis were exposed to R6 before fertilization, a W-O exposure resulted in a dose-related decrease in fertilization of P. lividus sperm (up to 250 microg/ml R6), whereas S. granularis sperm underwent a significant increase of fertilization rate at the highest R6 nominal levels (up to 25 microg/ml). Equivalent CYM or Cu-OCl levels were ineffective on sperm fertilization success in both species. The offspring of S. granularis sperm exposed to 25 microg/ml R6 showed a significant increase in larval malformations, which were not detected in the offspring of R6-exposed P. lividus sperm. Again, CYM or Cu-OCl was unable to exert any transmissible damage from sperm to the offspring in either species. The present study raises the case of possible discrepancies between toxicity of a technical mixture and of its analytical-grade components, also providing evidence for a loss of pesticide toxicity following dispersion in an environmental matrix such as sediment or soil. PMID- 11727792 TI - In vitro cytotoxicity evaluation of plastic biomedical devices. AB - Cytotoxic potential of four plastic biomedical devices (intravenous transfusion sets, IV sets; dextrose normal saline bottles, DNS bottles; Ringer lactate bottles, RL bottles; and Ryle's tubes) including 17 different brands was evaluated by investigating growth inhibition, percent survival, mitotic index and colony-forming ability (cfa) in L929, an adherent type mouse fibroblast cell line. Experimental sets were exposed with leachates of biomedical products in serum-free minimum essential medium (MEM) for 1 h at 37 degrees C in a CO2 incubator. After 1 h, medium was replaced with serum rich MEM containing essential amino acids and reincubated up to 96 h. Cells in serum-free MEM only were processed under identical conditions and served as the control. The leachates from all types of biomedical devices evaluated exhibited reduction in the growth and survival of the cell line in the first 12 h postexposure followed by their gradual recovery up to 96 h. A significant reduction in cell growth was apparent in the six brands of IV sets from 24 h onwards up to 36 h (59% growth inhibition). Though the cfa was also reduced in all the brands tested, the magnitude of reduction was less compared to growth inhibition. The results indicate that leachates of IV sets were more toxic compared to other biomedical devices screened, and growth inhibition assay was found to be more sensitive and suitable for cytotoxicity evaluation of biomedical devices. PMID- 11727793 TI - Stimulation of C32 and G361 melanoma cells using oleoyl acetyl glycerol and its effect on sulphur mustard cytotoxicity. AB - Epidermal melanocytes have a higher sensitivity to sulphur mustard (HD) compared with other skin cell types. This may be due to the enzymatic production of melanin precursors exerting an additional cytotoxic effect following HD depletion of the cellular protectant, GSH. Stimulation of the protein kinase C pathway in melanocytes is known to increase melanin production in melanocytes and melanoma cell lines. In order to investigate the role of pigment synthesis in HD toxicology, cultures of an unpigmented melanoma cell line (C32) and of a pigmented melanoma line (G361) were treated with the potent diacyl glycerol analogue, oleoyl acetyl glycerol (OAG), in order to determine if protein kinase C mediated increases in pigment production could increase sensitivity to subsequent HD exposure. Stimulation of C32 cells with OAG exerted a significant protective effect against the cytotoxic effects of HD. However, this was not due to increased melanin synthesis because this cell line cannot synthesize melanin pigments. The protective action observed is postulated to be due to modulation of protein kinase C activity. In contrast, stimulation of G361 melanoma cells with OAG resulted in an increased level of cytotoxicity upon subsequent exposure to HD. Protein kinase C controls several cellular pathways including checkpoints in the cell cycle, stalling the cell in G and promoting transition through the G2/M boundary. Given the genotoxic properties of HD, these two points in the cell cycle are important in determining the overall cytotoxic effect of HD. Control of the cell cycle by protein kinase C modulation and manipulation of melanin synthetic pathways may have therapeutic benefits. PMID- 11727794 TI - Chronicity of neurological features in ciguatera fish poisoning. AB - A 43-year-old woman complained of colicky abdominal pain, followed by numbness, myalgias, and muscle weakness in the four limbs after eating a grouper (Epinepheius spp.). She presented to our hospital 36 hours later with increased myalgias, muscle weakness, and malaise. On examination, the muscle power and sensation in her four limbs appeared to be normal. She was given an intravenous infusion of mannitol 20% (200 ml over 1 hour) and an intramuscular injection of diclofenac (75 mg). Her myalgias then improved and she was discharged. She presented to our hospital again 1 week later with poor appetite, malaise, numbness of the four limbs, and increased muscle weakness. On examination, the muscle weakness was more marked in the lower limbs (4+/5) than in the upper limbs (5-/5) and proximally than distally. She also had some difficulty in getting up from a squatting position. She was given another intravenous infusion of mannitol 20% (200 ml over 1 hour), following which there was subjectively slight improvement in her muscle weakness. Herplasma creatine phosphokinase level was normal. Electromyography performed 4 weeks later revealed no abnormalities. When she was reviewed 45 days after the consumption of the grouper, her muscle weakness and malaise had improved considerably. She could then stand up from a squatting position. However, mild impairment of finger grip was still present. Chronicity of neurological features in other reported cases (e.g., chronic fatigue, relapse of symptoms after exposure to ciguateric fish or alcohol, and peripheral neuropathy) may also indicate a lengthy persistence of ciguatoxins in the body. PMID- 11727795 TI - Decreased plasma and cerebrospinal fluid glutamine concentrations in a patient with bialaphos poisoning. AB - A 47-year-old Japanese woman undergoing maintenance hemodialysis (HD) was admitted to our hospital because of poisoning with the herbicide bialaphos. Respiratory arrest and loss of consciousness ensued rapidly, accompanied by convulsions and nystagmus. Treatment with HD and direct hemoperfusion, followed by HD alone, effectively removed bialaphos and its chief toxic metabolite (L AMPB) from the circulation (bialaphos decreased from 0.33 to < 0.05 microg/ml and L-AMPB from 14 to 0.86 microg/ml). The glutamate concentration improved gradually after the removal of bialaphos and L-AMPB from plasma (plasma glutamate concentration: 250.4 nmol/l on day 5 to 120.6 nmol/l on day 26). Decreased glutamine concentration in cerebrospinal fluid was demonstrated for the first time as well as in plasma, indicating glutamine synthetase inhibition not only in plants but also in humans by bialaphos poisoning. PMID- 11727796 TI - The hormesis challenge for environmental health regulators. PMID- 11727797 TI - The globalization of orthopedics. PMID- 11727798 TI - Radiologic case study. Exertional rhabdomyolysis. PMID- 11727799 TI - A modified anterior approach to the middle third of the humeral shaft. PMID- 11727800 TI - Rotational alignment of the tibial component in total knee arthroplasty. AB - This study compares two different methods of aligning the tibial component in a total knee arthroplasty, one by aligning the component with the border between the medial one third and lateral two thirds of the tibial tubercle and the other by allowing the component to seek its own position as the knee is taken through a range of motion. The first technique emphasizes the need to maintain an appropriate quadriceps angle for optimum patellar tracking and the second method optimizes femorotibial kinematics. The two methods suggest different positions for the tibial component, with the tibial component internally rotated an average of 5.6 mm from the border between the medial one third and lateral two thirds of the tibial tubercle when allowed to seek its own rotation. Surgeons must be aware of the choices they are making when selecting a method of aligning the tibial component. The prosthesis used in this study had a relatively high level of rotational constraint. The situation is somewhat different in a less constrained design. PMID- 11727801 TI - Mid-term results in total hip arthroplasty with the basic noncemented hip prosthesis. AB - The Basic (IQL-Biomet, Valencia, Spain) hip prosthesis was used in 430 implant procedures, of which 417 had mean follow-up of 6 years. Radiographic follow-up was performed at 1, 3, 6, and 12 months postoperatively, and annually thereafter. The average Harris hip score improved from 35.7 preoperatively to 90.9 postoperatively. Clinical results after 2 years of follow-up showed 6% thigh pain, which decreased to 2% at 6 years. Radiographic evaluation demonstrated 22 cases of calcar resorption, 14% of which had distal pedestal formation. Heterotopic ossification was noted radiographically in 36%. These mid-term results obtained with the Basic prosthesis are similar to the findings of other series and warrant its use. PMID- 11727802 TI - Hand infections in patients with diabetes mellitus. AB - Fifty diabetic patients with hand infections were studied retrospectively. The cause of infection varied, and the infections were divided into two groups: superficial infections and abscesses. The amputation rate was 14%. Only patients who presented with an abscess required amputation, and of those patients with abscesses, 17.5% required amputation. Eighteen of 38 cultured infections were polymicrobial, 4 contained Enterococcus, 11 contained gram-negative bacteria, and 3 contained anaerobic organisms. Bacteriologic culture analysis initiated a two drug protocol: ampicillin with sulbactam (Unasyn; Pfizer, New York, NY) or piperacillin with tazobactam (Zosyn; Lederle, Pearl River, NY) and gentamicin (renal adjusted). PMID- 11727803 TI - Osteoid osteoma and osteoblastoma of the sacrum. AB - This retrospective study examined 10 patients with osteoid osteoma and 11 patients with osteoblastoma localized in the sacrococcyx. In the sacrum, the diagnosis was delayed compared to other sites. Curettage through a posterior approach is the treatment of choice. Radiotherapy as well as embolization of feeding arteries may be used for the most aggressive lesions. Prognosis is generally good with a low incidence of local recurrence (<10%). PMID- 11727804 TI - Three-dimensional computerized selection of hip prostheses in patients with congenital dislocated hips. AB - This study assessed the effectiveness of the combined use of computed tomography (CT) and computer-aided design (CAD) in the preoperative evaluation and implant selection in 20 patients (20 hips) with congenital dislocation of the hip who were scheduled to undergo total hip arthroplasty. Computerized selection of the femoral implant with optimum fit and fill was made after a three-dimensional reconstruction of the femoral canal using CT data and CAD. Implantation of all sizes of 5 noncemented and 2 cemented femoral implants was simulated using CATIA software (IBM, Kingstone, NY). When patients underwent surgery, 18 of 20 preselected prostheses agreed by type and size with the prostheses implanted. The remaining 2 preselected implants agreed by type only. In patients with dislocated and dysplastic hips, combined use of CT and CAD allows effective preoperative planning by providing the surgeon with vital information about the proximal femoral canal geometry and the possible femoral implant with optimum fit and fill to be used. PMID- 11727805 TI - Predicting the rotationally neutral state of the femur by comparing the shape of the contralateral lesser trochanter. AB - The rotationally neutral state of the femur was measured by comparing the shape of the contralateral lesser trochanter in 100 adults. Three observers predicted a position presumed to be neutral by comparing the shape of the neutrally positioned contralateral lesser trochanter previously recorded by C-arm image intensification. The mean (SD) angular discrepancy between the neutral point and the predicted angle was 3.4 degrees (+/- 2.4) without interobserver variation (P = .87). Seventy-seven percent indicated the value within 4 degrees, and none of the observations exceeded 15 degrees. These findings suggest the outline of the contralateral lesser trochanter may be a useful reference during surgical treatment of femoral fractures to minimize rotational deformity. PMID- 11727806 TI - Radiographic evaluation of transverse sacral fractures. AB - This study clarifies the pattern of fracture lines and facilitates diagnosis of transverse sacral fracture on plain radiographic images. Eight cadaveric sacra were used in this study. A U-shaped transverse sacral fracture at the S2-S3 level was created in all specimens. The fracture line was marked by painting with radio opaque material and solder metal wires. The following radiographic views were taken: anteroposterior, lateral, AP with 35 degrees cephalad orientation, and inlet view. A double shadow in the upper sacral area can be identified in the plain AP view. As a consequence of the fracture, there are changes in the orientation of the planes of the foramina from the coronal to, more or less, axial plane. Anteroposterior with 35 degrees cephalad orientation radiographs provide good assessment for evaluation of the transverse sacral fracture. PMID- 11727807 TI - Classification of heterotopic ossification about the elbow. PMID- 11727808 TI - Vascular compromise due to supracondylar fracture of the femur after total knee arthroplasty. PMID- 11727809 TI - Limb lengthening after limb sparing for osteogenic sarcoma of the proximal humerus. PMID- 11727810 TI - Fungal osteomyelitis of the distal femoral epiphysis. PMID- 11727811 TI - Giant-cell tumor of the capitate. PMID- 11727812 TI - Aneurysmal bone cyst associated with fibrous dysplasia. PMID- 11727813 TI - The role of arthroscopic intervention for symptomatic total knee arthroplasty. PMID- 11727814 TI - New antibiotics. PMID- 11727815 TI - Zentralblatt fur bakteriologie--100 years ago an outbreak of fowl plague in Tyrol in 1901. PMID- 11727816 TI - Dendritic cells as vectors for vaccination against infectious diseases. AB - Antigen presentation by dendritic cells (DCs) is critical for the induction of a specific immune response. The immunotherapeutic potential of antigen-pulsed DCs for the treatment of cancer has been confirmed in a number of experimental tumor models and in several preclinical trials. Recent advances in our understanding of the interaction of microbial pathogens with DCs have provided the basis to explore DCs as vaccine carriers for the induction of protective immune responses to infections. Support for this strategy comes from animal studies demonstrating that DCs, after ex vivo loading with microbial antigens, confer protection against microbial challenges in vivo. This may have important implications for the development of novel strategies for prophylactic or therapeutic immunizations against various microbial pathogens. PMID- 11727817 TI - Pathogenicity of Legionella pneumophila. AB - The bacterium Legionella pneumophila is the principal etiologic agent of Legionnaires' disease, a form of lobar pneumonia. Ubiquitous in aquatic environments, the gram-negative Legionella organism is a facultative, intracellular parasite of protozoa. The pathogenesis of legionellosis is largely due to the ability of L. pneumophila to invade and grow within alveolar macrophages, and it is widely believed that this ability results from a prior adaptation to intracellular niches in nature. Indeed, intracellular legionellae display a remarkable capacity to avoid endosomal and lysosomal bactericidal activities and to establish a unique replicative phagosome. In recent years, much progress has been made toward identifying the bacterial factors that promote intracellular infection and virulence. Surface structures that enhance infection include LPS, flagella, type IV pili, an outer membrane porin, and the Mip propyl proline isomerase. Both type II and type IV protein secretion systems are critical for L. pneumophila pathogenesis. Whereas the type II (Lsp) system secretes a collection of degradative enzymes, the type IV (Dot/Icm) system likely exports effector proteins that are especially critical for trafficking of the Legionella phagosome. In addition to facilitating pilus formation and type II secretion, the inner membrane prepilin peptidase (PilD) of L. pneumophila appears to mediate a third, potentially novel pathway that is operative in the mammalian host. Periplasmic and cytosolic infectivity determinants include a catalase peroxidase and the HtrA and Hsp60 stress-response proteins. The stationary phase response and the iron acquisition functions of L. pneumophila also play key roles in pathogenesis, as do a number of other loci, including the pts, mil and enh genes. PMID- 11727818 TI - A point mutation in the active site of Legionella pneumophila O-acetyltransferase results in modified lipopolysaccharide but does not influence virulence. AB - The majority of clinical isolates of Legionella pneumophila serogroup 1 produce lipopolysaccharide (LPS) that reacts with monoclonal antibody (MAb) 3/1. By using a negative cell sorting method, we isolated a spontaneous LPS mutant from L. pneumophila serogroup 1 strain Corby that lost reactivity with this MAb. The mutant contained a single nucleotide exchange in position 169 of the lag-1 gene that encodes an O-acetyltransferase that is responsible for O-acetylation of the L. pneumophila O-repeat unit (legionaminic acid). This mutation resulted in a single amino acid exchange in a highly conserved motif present in many O acetyltransferase-like proteins. RT-PCR analysis revealed that the mutant lag-1 gene was transcribed, but the resulting protein lacked O-acetyltransferase activity. Chemical analysis of the mutant LPS revealed that it lacked 8-O-acetyl groups in legionaminic acid. In addition, the mutant failed to produce high molecular-weight long-chain O-polysaccharide. Complementation of the mutant with the wild-type lag-1 gene restored reactivity with MAb 3/1 and the chemical structure of the wild-type LPS. Strain Corby and its MAb 3/1-negative mutant were indistinguishable in their serum resistance characteristics, and in uptake and intracellular multiplication in Acanthamoeba castellanii and macrophages. PMID- 11727819 TI - Overexpresssion of a Legionella pneumophila homologue of the E. coli regulator csrA affects cell size, flagellation, and pigmentation. AB - Legionella pneumophila is an inhabitant of the aquatic environment and the causative agent of a bacterial pneumonia. We identified the presence of an L. pneumophila homologue of csrA of E. coli and rsmA of Erwinia carotovora, genes which regulate gene expression by destabilising mRNA and which have been shown to relate to environmental fitness and pathogenicity. The Legionella csrA was able to complement a csrA-negative mutant of E. coli. Overproduction of csrA in L. pneumophila lead to a reduction of flagellation and pigmentation and an increase in bacterial cell size. csrA overproduction was associated with a reduction of fliA and flaA transcripts. This suggests that similar to E. coli and Erwinia, L. pneumophila csrA is a regulator of gene expression and may contribute to the capability of the pathogen to rapidly adapt to changing environments. PMID- 11727820 TI - Analysis of transcriptional control mechanisms of capsule expression in Neisseria meningitidis. AB - The major virulence factor which contributes to the survival of Neisseria meningitidis in the blood stream and the cerebrospinal fluid is the capsular polysaccharide. Expression of the capsule genes of N. meningitidis serogroups B, C, W-135 and Y is controlled by an intergenic region separating the capsule biosynthesis operon (siaA-D) and the capsule transport operon (ctrA-D). To further investigate capsule expression in N. meningitidis we amplified and sequenced the intergenic region of 42 meningococcal isolates of different serogroups. Sequence variations were found mainly in a repeat region preceding the siaA start codon. Correlation between sequence variation and serogroup could not be observed. To measure the transcriptional and translational activity of the respective intergenic regions we performed transcriptional and translational fusions with the lacZ gene integrated into the chromosome of N. meningitidis. Sequence variations preceding the siaA start codon had no effect on beta galactosidase activity. Different in vitro growth conditions such as temperature, glucose concentration, osmolarity, pH and iron concentration also did not influence beta-galactosidase activity. Sequential deletions of the intergenic region showed that an Up-like element adjacent to the predicted -35 box is necessary for full transcriptional activity. The deletion of the untranslated region preceding the siaA start codon led to a threefold higher beta galactosidase activity compared with the full-length construct suggesting that the respective region may be involved in capsule regulation. PMID- 11727821 TI - Virulence-associated genes in avian pathogenic Escherichia coli (APEC) isolated from internal organs of poultry having died from colibacillosis. AB - Escherichia coli infections are responsible for significant losses in the poultry industry in many parts of the world. The pathogenesis and the role of virulence factors are not yet totally elucidated. We, therefore, examined 150 E. coli strains isolated from visceral organs of poultry having died from colibacillosis for the presence of virulence-associated genes by PCR. The E. coli strains were investigated for the presence of a total of 17 virulence-associated genes described for diarrheagenic (stx1/2, eae, hlyEHEC, estl, eltI, astA, cdtb), septicemic (hlyA, papC, cnf1/2, fyuA, irp2) and avian pathogenic E. coli (APEC; iucD, tsh, fimC, and hlyE as well as stx2f). Seven genes were significantly distributed among APEC strains, while most of the other investigated genes could be demonstrated only sporadically or not at all. FimC (Type I fimbriae) was detected with the highest prevalence in 92.7% of the isolates. Most of the strains harboring iucD (88.7%) also gave positive results for tsh (85.3%). Genes fyuA (ferric yersiniabactin uptake) (66.0%) and irp2 (iron-repressible protein) (68.0%), necessary for Yersinia to acquire iron in the mouse infection model, were regularly detected in combination. Moreover, we found papC (pyelonephritis associated pili) in 30.0% and astA (enteroaggregative heat stable toxin) in 17.3% of the field strains. A significant amount of strains (57.3%) harbored a combination of iucD, tsh, fimC, fyuA and irp2 virulence-associated genes, presumably rendering these strains particularly virulent. These findings provide novel insights into the presence and distribution of virulence-associated genes in avian pathogenic E. coli field strains, which will help to more comprehensively characterize APEC in future epidemiological studies. It is assumed that the existence of two iron acquisition systems points towards their important role in virulence. Furthermore, we suggest that characterization of the respective phenotypes in infection models will provide substantial information to better understand the pathogenesis of colibacillosis in poultry. PMID- 11727822 TI - Genotypic and serotypic stability of Campylobacter jejuni strains during in vitro and in vivo passage. AB - The stability of four typing methods and the sero- and genotypic stability of three Campylobacter jejuni strains were evaluated after subculturing 50 times in triplicate and after colonising mice for up to 26 days. The employed methods were Penner heat-stable serotyping; automated ribotyping (RiboPrinting) using HaeIII restriction enzyme; pulsed-field gel electrophoresis (PFGE) using SmaI, SalI and KpnI; and random amplified polymorphic DNA analysis (RAPD) using primers 1254, 1281 and HLWL85. No changes in any of the DNA profiles or in the reactions to heat-stable antigens were identified among these strains after the in vitro and in vivo passages. However, one isolate became untypeable with RAPD after passage in one of the mice. In addition, eleven other C. jejuni strains of four different serotypes were subcultured ten times to screen for instability. Neither of these showed instability using PFGE and serotyping. Furthermore, three of four strains previously identified as unstable, showed to consist of mixed cultures, which explains the reported profile changes. The results indicate that the applied typing methods are reliable and applicable for typing of Campylobacter isolates from different sources over time, and that many C. jejuni strains are genetically stable as tested by these methods. PMID- 11727823 TI - Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. AB - Chronic lung infection with Pseudomonas aeruginosa is primarily responsible for pulmonary deterioration of cystic fibrosis patients. The purpose of this study was to type the P. aeruginosa isolates collected sequentially from cystic fibrosis patients, chronically colonized with P. aeruginosa, by random amplified polymorphic DNA fingerprinting-PCR (RAPD-PCR). Sequential P. aeruginosa isolates (n: 130) that had been collected from 20 CF patients over at least 9 years were investigated. The isolates were analyzed by RAPD-PCR using two arbitrary primers. Antimicrobial susceptibility testing of all isolates was performed by the disc diffusion method. RAPD-PCR typing demonstrated that strains dissimilar in colony morphotype and of different antibiotic susceptibility patterns could be of the same genotype. Some CF patients were colonized with a rather constant P. aeruginosa flora, with strains of different phenotypes but of one genotype. However, some patients may be colonized with more than one genotype. The results also demonstrated that there might be a risk of cross-colonization between CF patients followed-up at the same center. PMID- 11727824 TI - Protein-DNA interaction and CpG methylation at rep*/vIL-10p of latent Epstein Barr virus genomes in lymphoid cell lines. AB - The viral interleukin-10 promoter (vIL-10p), overlapping the rep* element in the Epstein-Barr virus (EBV) genome, is a promoter element active mostly in the late phase of the lytic cycle and immediately upon infection of B cells. rep* was, through transfection experiments with small plasmids, characterised as a cis element supporting oriP replicative function. In this study, in vivo protein binding and CpG methylation at rep*/vIL-10p were analysed in five cell lines that harbour strictly latent EBV genomes. Contrary to the invariably unmethylated dyad symmetry element (DS) of oriP, rep*/vIL-10p was highly methylated and showed only traces of protein binding in all examined cell lines. This result is in agreement with vIL-10p being an inactive promoter of EBV genomes, and makes it less likely that rep* functions as a replicative element of latent EBV genomes. PMID- 11727825 TI - In vitro and in vivo processing of cyanelle tmRNA by RNase P. AB - Ribonuclease P, the ubiquitous endonuclease required for generating mature tRNA 5' ends, is a ribonucleoprotein in most organisms and organelles, with the exception of mitochondria and chloroplasts of multicellular organisms. The cyanelle of the primitive alga Cyanophora paradoxa is the only photosynthetic organelle where the ribonucleoprotein nature of this enzyme has been functionally proven. tmRNA is another highly structured RNA: it can be aminoacylated with alanine, which is then incorporated into a tag peptide encoded on the same RNA molecule. This dual-function RNA has been found in bacteria, and its gene is also present in mitochondria and plastids from primitive organisms. Since nothing is known about the expression of this RNA in organelles, we have performed processing studies and determined the promoter of cyanelle pre-tmRNA. This RNA is transcribed as a precursor molecule in vivo. Synthetic transcripts of cyanelle pre-tmRNA, including or lacking the mature 3' CCA-end, are efficiently and correctly processed in vitro by bacterial RNase P ribo- and holoenzymes and by the homologous cyanelle RNase P. In addition to these experimental data, we propose a novel secondary structure model for this organellar tmRNA, which renders it more similar to its bacterial counterpart. PMID- 11727826 TI - A eubacterial origin for the human tRNA nucleotidyltransferase? AB - tRNA CCA-termini are generated and maintained by tRNA nucleotidyltransferases. Together with poly(A) polymerases and other enzymes they belong to the nucleotidyltransferase superfamily. However, sequence alignments within this family do not allow to distinguish between CCA-adding enzymes and poly(A) polymerases. Furthermore, due to the lack of sequence information about animal CCA-adding enzymes, identification of corresponding animal genes was not possible so far. Therefore, we looked for the human homolog using the baker's yeast tRNA nucleotidyltransferase as a query sequence in a BLAST search. This revealed that the human gene transcript CGI-47 (#AF151805) deposited in GenBank is likely to encode such an enzyme. To identify the nature of this protein, the cDNA of the transcript was cloned and the recombinant protein biochemically characterized, indicating that CGI-47 encodes a bona fide CCA-adding enzyme and not a poly(A) polymerase. This confirmed animal CCA-adding enzyme allowed us to identify putative homologs from other animals. Calculation of a neighbor-joining tree, using an alignment of several CCA-adding enzymes, revealed that the animal enzymes resemble more eubacterial ones than eukaryotic plant and fungal tRNA nucleotidyltransferases, suggesting that the animal nuclear cca genes might have been derived from the endosymbiotic progenitor of mitochondria and are therefore of eubacterial origin. PMID- 11727827 TI - Interaction of thymidylate synthase with the 5'-thiophosphates, 5' dithiophosphates, 5'-H-phosphonates and 5'-S-thiosulfates of 2'-deoxyuridine, thymidine and 5-fluoro-2'-deoxyuridine. AB - New analogs of dUMP, dTMP and 5-fluoro-dUMP, including the corresponding 5' thiophosphates (dUMPS, dTMPS and FdUMPS), 5'-dithiophosphates (dUMPS2, dTMPS2 and FdUMPS2), 5'-H-phosphonates (dUMP-H, dTMP-H and FdUMP-H) and 5'-S-thiosulfates (dUSSO3, dTSSO3 and FdUSSO3), have been synthesized and their interactions studied with highly purified mammalian thymidylate synthase. dUMPS and dUMPS2 proved to be good substrates, and dTMPS and dTMPS2 classic competitive inhibitors, only slightly weaker than dTMP. Their 5-fluoro congeners behaved as potent, slow-binding inhibitors. By contrast, the corresponding 5'-H-phosphonates and 5'-S-thiosulfates displayed weak activities, only FdUMP-H and FdUSSO3 exhibiting significant interactions with the enzyme, as weak competitive slow binding inhibitors versus dUMR The pH-dependence of enzyme time-independent inhibition by FdUMP and FdUMPS was found to correlate with the difference in pKa values of the phosphate and thiophosphate groups, the profile of FdUMPS being shifted (approximately 1 pH unit) toward lower pH values, so that binding of dUMP and its analogs is limited by the phosphate secondary hydroxyl ionization. Hence, together with the effects of 5'-H-phosphonate and 5'-S-thiosulfate substituents, the much weaker interactions of the nucleotide analogs (3-5 orders of magnitude lower than for the parent 5'-phosphates) with the enzyme is further evidence that the enzyme's active center prefers the dianionic phosphate group for optimum binding. PMID- 11727828 TI - The MKK6/p38 mitogen-activated protein kinase pathway is capable of inducing SOCS3 gene expression and inhibits IL-6-induced transcription. AB - In this study we show that activation of p38MAPK by IL-6 acts as an inhibitory signal on IL-6-mediated activation of STAT and the alpha2-macroglobulin promoter. We analyzed the role of MKK6/p38MAPK for IL-6 signal transduction and transcriptional activation of the suppressor of cytokine signaling (SOCS) 3 promoter. Pretreatment of cells with the p38MAPK-specific inhibitor SB202190 downregulates the induction of SOCS3-mRNA expression by IL-6. Accordingly, overexpression of a constitutively active MKK6 in HepG2 cells enhanced basal activity or IL-6-induced transcriptional activation of a SOCS3 promoter reporter construct, whereas overexpression of a dominant negative mutant of MKK6 downregulated the IL-6-mediated activation of the SOCS3 promoter. These data indicate that p38MAPK-activation is crucial for IL-6-induced SOCS3 expression and downregulation of IL-6-mediated gene induction. PMID- 11727830 TI - Major vault protein is a substrate of endogenous protein kinases in CHO and PC12 cells. AB - Major vault protein (MVP) is the predominant member of a large cytosolic ribonucleoprotein particle, termed vault. We have previously shown that MVP derived from electric ray electric organ becomes phosphorylated by protein kinase C in vitro and by tyrosine kinase in vivo. Here we show that MVP from two mammalian cell lines (CHO and PC12 cell) becomes highly phosphorylated by endogenous protein kinases in cell-free systems. The susceptibility to protein kinases differs substantially from those observed in MVP derived from electric organ. Phosphorylation of MVP depends on the presence of Mg2+ and can be inhibited by the chelating agent EDTA. Inhibitors of casein kinase II attenuate the phosphorylation of MVP. In contrast to CHO cells, addition of recombinant casein kinase II enhances the phosphorylation of MVP in PC12 cells. Endogenous kinase activity is of particulate nature and copurifies with vault particles. Immuno-affinity purified vaults containing recombinant tagged MVP expressed in CHO cells reveal no autophosphorylation, suggesting that protein kinase activity is not an intrinsic property of vaults. Our results suggest that cell-specific phosphorylation of MVP may play a critical role in vault function. PMID- 11727829 TI - Identification of a gephyrin-binding motif in the GDP/GTP exchange factor collybistin. AB - The brain-specific GDP/GTP exchange factor collybistin interacts with the receptor-anchoring protein gephyrin and activates the Rho-like GTPase Cdc42, which is known to regulate actin cytoskeleton dynamics. Alternative splicing creates two collybistin variants, I and II. In coexpression experiments, collybistin II has been shown to induce the formation of submembraneous gephyrin aggregates which cluster with hetero-oligomeric glycine receptors (GlyRs). Here we identified residues critical for interaction with gephyrin in the linker region between the SH3 and the DH domains of collybistin. Respective collybistin deletion mutants failed to bind gephyrin upon coexpression in heterologous cells, in GST pull-down assays and in the yeast two-hybrid system. Site-directed mutagenesis revealed polar amino acid residues as essential determinants of gephyrin binding. Furthermore, in vitro gephyrin bound simultaneously to both collybistin and the GlyR beta-subunit binding motif. Our data are consistent with collybistin-gephyrin interactions occuring during inhibitory postsynaptic membrane formation. PMID- 11727831 TI - Identification of receptors for prothymosin alpha on human lymphocytes. AB - Prothymosin alpha (ProTalpha) is a highly conserved and widely distributed protein whose physiological functions remain elusive. In previous work we identified high and low affinity-binding sites for ProTalpha in lymphoid cells. Here we demonstrate, by affinity cross-linking and affinity chromatography, the existence of three binding partners (31, 29, and 19 kDa) for ProTalpha in the membrane of PHA-activated lymphoblasts. These surface molecules possess the expected affinity and specificity for a ProTalpha receptor. Examination of the expression of this complex of molecules by flow cytometry reveals that they bind ProTalpha in a specific and saturable way. In addition, the distribution of the receptor on the cell surface was studied by fluorescence microscopy; a cap-like structure at one of the poles of the cells was identified. These results represent a new and promising approach in the research on ProTalpha, opening the way toward the understanding of the molecular mechanism of action of this protein. PMID- 11727832 TI - Tissue kallikrein KLK1 is expressed de novo in endothelial cells and mediates relaxation of human umbilical veins. AB - Bradykinin released by the endothelium is thought to play an important local role in cardiovascular regulation. However, the molecular identity of endothelial proteases liberating bradykinin from its precursors remained unclear. Using RT PCR and Southern blotting techniques we detected mRNA for tissue kallikrein (KLK1) in human umbilical vein endothelial cells and in bovine aortic endothelial cells. Protein expression was confirmed by precipitation of KLK1 from lysates of endothelial cells pre-labeled with [35S]-cysteine/methionine. Partial purification of tissue kallikrein from total endothelial cell extracts resulted in a protein triplet of about 50 kDa in Western blots using specific anti-KLK1 antibodies. The immunodetection of tissue kallikrein antigen in the fractions from ion exchange chromatography correlated with the presence of amidolytic tissue kallikrein activity. Stimulation of endothelial cells with angiotensin II (ANG-II), which recently has been shown to activate the vascular kinin system and to cause vasodilation, resulted in the release of bradykinin and kallidin. ANG-II dependent relaxation of pre-constricted rings from human umbilical veins was abolished in the presence of a specific tissue kallikrein inhibitor. We conclude that endothelial cells de novo express significant amounts of tissue kallikrein, which likely serves in the local generation of vasoactive kinins. PMID- 11727833 TI - Secretion of gelatinases and activation of gelatinase A (MMP-2) by human rheumatoid synovial fibroblasts. AB - In monolayer cultures human rheumatoid synovial fibroblasts (HRSF) secrete gelatinase A (MMP-2) and, unlike other human fibroblasts, to a minor extent also gelatinase B (MMP-9) as inactive proenzymes. In this regard HRSF resemble the fibrosarcoma cell line HT-1080. Unlike HT-1080, however, HRSF do not increase the secretion of MMP-9 in response to phorbol-12-myristate-13-acetate. This indicates that in HRSF the protein kinase C pathway for an enhanced MMP-9 secretion is inactive. None of the substances used in our study increased MMP-9 secretion, but some of them inhibited MMP-9 secretion. The secretion of MMP-2 could not be enhanced either, not even by dbcAMP, which has been reported to be effective in Sertoli and peritubular cells. Activation of MMP-2 in HRSF could be induced by treatment with concanavalin A (ConA) or cytochalasin D, as was shown for other cell types. This activation was not accompanied by a significant change in the amount of secreted TIMP-1 and TIMP-2. In contrast to reports on human skin fibroblasts, however, the activation of MMP-2 could not be induced in HRSF by treatment of the cells with monensin or sodium orthovanadate. Moreover, monensin was shown to act as an inhibitor of ConA- or cytochalasin D-mediated activation. Additionally, and in contrast to a report on a rat fibroblast cell line, MMP-2 activation is not mediated via the MAP kinase pathway in HRSF: PD 98059, a specific inhibitor of MAP kinase kinase, did not inhibit the activation of MMP-2. Similarly ineffective were PD 169316, an inhibitor for p38 MAP kinase, other inhibitors for protein kinases as lavendustin A, Go 6983, wortmannin, rapamycin, as well as the protein tyrosine kinase inhibitors herbimycin A and genistein. Only staurosporin, a broad spectrum inhibitor of protein kinases, and the ionophores monensin and A 23187 effectively inhibited MMP-2 activation in HRSF. Our results demonstrate that MMP-2 can be activated by quite different pathways, and that different cells, even when belonging to the fibroblast family, do not necessarily use the same activating pathways. PMID- 11727834 TI - Ferredoxins from the archaeon Acidianus ambivalens: overexpression and characterization of the non-zinc-containing ferredoxin FdB. AB - Two ferredoxin genes, fdA and fdB, from the extremely thermoacidophilic crenarchaeon Acidianus ambivalens have been sequenced; the sequences share 86% similarity. Whereas the deduced protein sequence of the ferredoxin FdA clearly contains a zinc-binding motif, the corresponding sequence of the FdB is devoid of this motif. Thus far, only the zinc-containing ferredoxin, FdA, from A. ambivalens has been chemically and functionally characterized from its native source. Using RT-PCR and Northern blot analysis, we show that both ferredoxins are expressed by A. ambivalens under either anaerobic or aerobic growth conditions. The zinc-free ferredoxin, FdB, was overexpressed in E. coli and purified to homogeneity. Using EPR spectroscopy, we could demonstrate that FdB contains one [3Fe-4S](1+/0) and one [4Fe-4S](2+/1+) cluster. The reduction potential of the [3Fe-4S](1+/0) cluster was determined as -235+/-10 mV, at pH 6.5, by EPR-monitored redox titration. The high melting temperature of 108+/-2 degrees C of FdB determined by CD spectroscopy reveals that it is not the binding of the Zn2+ that induces the extreme thermostability of these ferredoxins. PMID- 11727836 TI - The N-terminal part of recombinant human tear lipocalin/von Ebner's gland protein confers cysteine proteinase inhibition depending on the presence of the entire cystatin-like sequence motifs. AB - Human Tear Lipocalin/von Ebner's gland protein (TL) is a member of the lipocalin superfamily. The protein is secreted by a number of serous glands and tissues and is overproduced under conditions of stress, infection and inflammation. In addition to its typical affinity for lipophilic ligands it was recently found to be able to inhibit cysteine proteinases [van't Hof et al., J. Biol. Chem. 272 (1997), 1837-1841], probably due to the presence of amino acid motifs resembling the papain binding domains of family 2 cystatins. In this work we have used a recombinant protein to confirm the results obtained with native TL. The inhibitory activity of the recombinant protein against papain was dependent on the ratio of papain and TL. At higher papain concentrations, the N-terminal sequence of TL was cleaved off by the protease, indicating that it can act in an inhibitor- or a substrate-like mode. This behaviour resembles that observed with certain chicken cystatin mutants. Using a recombinant TL mutant we found that the two Leu residues (Leu4-Leu5) contained within the first cystatin-like motif are absolutely essential for the inhibitory activity. These results were supported by experiments using a recombinant form of the corresponding pig von Ebner's gland protein (VEGp). This protein, which does not possess a fully conserved first cystatin-like motif, is unable to inhibit papain. PMID- 11727835 TI - Isolation and characterization of a trypsin-like protease from Trichoderma viride. AB - A serine endopeptidase with a molecular mass of 25 kDa has been purified from the culture filtrate of Trichoderma viride to electrophoretic homogeneity. The isoelectric point was determined at 7.3. Two carboxyl sites at Arg22 and Lys29 of the oxidized insulin B-chain were cleaved, and peptidyl-p-nitroanilide substrates with Lys or Arg at the P1 position were also hydrolyzed by the enzyme. These results suggest that the specificity of T. viride protease is similar to that of trypsin. However, the hydrolytic activity toward casein of T. viride protease was less than that of porcine trypsin. The amino-terminal sequence of the enzyme protein is similar to that of bovine trypsin. It seems that the trypsin of T. viride is a protease which is promising for the substitution of animal trypsin in the food industry and in medicine at this stage. PMID- 11727837 TI - Treatment of rheumatoid arthritis in the third millennium. AB - Immense progress has been made in recent years in the management of patients with autoimmune diseases, such as rheumatoid arthritis (RA). This progress is largely owed to the advances in our understanding of cellular and molecular mechanisms involved in the pathogenesis of inflammation and autoimmunity and the substantially improved knowledge of molecular biology. In this editorial review, we delineate recent developments in the treatment of RA with a focus on biological therapeutics. we outline the concerns and open questions risen by the current studies and we portray future expectations based on the recent observations from those trials with biological treatment interventions. PMID- 11727838 TI - Clinical investigation of methotrexate in the treatment of ankylosing spondylitis. AB - OBJECTIVES: To investigate the efficacy and possible side effects of Methotrexate (MTX) treatment in patients with ankylosing spondylitis. METHODS: A total of 51 patients with a diagnosis of AS according to NewYork criteria were randomly distributed into 2 groups. In Group I (n = 25) only Naproxen (1000 mg/day) and in Group II (n = 26) oral MTX (7.5 mg/week) combined with Naproxen were given for 12 months. The patients were evaluated at 3, 6, and 12 months after the start of the treatment according to a number of clinical parameters using objective and subjective variables. Data were analysed using ki-square and Mann-Whitney U tests. RESULTS AND CONCLUSIONS: The comparison of the values using the percent changes and difference scores with respect to pretreatment values showed significant improvement in the "global evaluation of the physician" in Group II compared to Group I while no other parameter was found to be significantly different between the two groups. Those results have led us to conclude that a combination of MTX with Naproxen did not prove to be superior to Naproxen-alone treatment in terms of statistical significance. Further trials with MTX employing larger doses and different patient populations, mainly comprised of peripheral arthritis are necessary for delineation of the role of MTX in AS treatment. PMID- 11727840 TI - Intra-articular glucocorticoid treatment of the rheumatoid wrist. An ultrasonographic study. AB - Fifty consecutive patients with RA and clinical wrist synovitis were randomly allocated to either receive an injection of glucocorticoid into the radiocarpal joint or have the same amount of drug divided into the radiocarpal and midcarpal joints. Inferior radioulnar joint synovitis was treated with extra steroid injection only in the latter group. Patient's and doctor's assessments as well as ultrasonography improved significantly at month 3 in both groups. A statistical difference between the groups was found in the midcarpal joint measurement favouring the extra midcarpal injection. Ten wrists (20%) were normal when assessed with ultrasound at month 3 while 34 wrists (68%) were normal on clinical assessment. PMID- 11727839 TI - Prednisolone maintenance dose in relation to starting dose in the treatment of polymyalgia rheumatica and temporal arteritis. A prospective two-year study in 273 patients. AB - OBJECTIVE: To describe the maintenance dose and annual cessation rate of oral corticosteroids in relation to the starting dose in patients with polymyalgia rheumatica (PMR) and temporal arteritis (TA). METHODS: A prospective two-years observational study of 273 patients with PMR and TA followed by rheumatologists. RESULTS: Mean daily maintenance dose of prednisolone during the first and second year was 5.7 mg and 4.3 mg for PMR, 6.6 mg and 4.1 mg for TA, and 8.3 mg and 4.7 mg for PMR with TA. There was a strong association between the initial dose and maintenance dose. The rate of steroid cessation after two years in PMR was 24%, in TA 16%, and in PMR with TA 5%. CONCLUSION: Low initial dose of prednisolone is associated with low maintenance dose. This is important as the majority of patients with PMR and TA will be treated for more than two years. PMID- 11727841 TI - Renal abnormalities in a population of patients with psoriatic arthritis. AB - OBJECTIVE: To investigate the prevalence of and to identify predictive factors for renal abnormalities in patients with psoriatic arthritis (PsA). METHODS: 73 patients with PsA were consecutively examined by laboratory analyses and clinically for joint manifestations. Renal function was estimated by creatinine clearance and urinary albumin. RESULTS: 17 (23.3%) of the patients had renal abnormalities as defined by creatinine clearance below the lower cut off of normal distribution (mean - 2 SD) and/or urinary excretion of albumin more than 25 mg/24 h. These patients were significantly older at the time of the study, older at joint disease onset, had longer skin disease duration, increased serum levels of beta2-microglobulin, and higher incidence of increased ESR and/or CRP levels. Increased ESR/CRP levels had significantly predictive value in multivariate analysis. CONCLUSIONS: In this study subclinical renal abnormalities was a prevalent finding. Predictive factor was inflammatory activity measured by laboratory variables. There were no predisposing effects of NSAID or DMARD therapy. PMID- 11727842 TI - Transesophageal and transthoracic echocardiography and Doppler-examinations in systemic lupus erythematosus. AB - OBJECTIVE: To search for cardiac abnormalities in systemic lupus erythematosus (SLE). METHODS: 35 patients examined by 2-D transthoracal Doppler and transesophageal echocardiography. RESULTS: Mitral and aortic valve abnormalities were seen in 12 patients (34%) respectively, and occurred altogether in 16 patients (46%). They were in general significantly associated with longer disease duration, but not with anticardiolipin antibodies (aCL), disease activity, or any other variable, except for time on corticosteroids. which was significantly longer in patients with aortic valve calcifications. CONCLUSION: Valve masses and valve thickening--often in combination--are the most frequent structural findings in SLE, occurring more often on the aortic than on the mitral valves. Factors other than antiphospholipid antibodies, medication, hypertension, or coronary heart disease seem to be responsible for this phenomenon. Drugs that modulate inflammation in endo- and pericardial tissue may, at least in part, be responsible for the observed mitral valve calcifications and pericardial fibrosis. PMID- 11727843 TI - Longitudinal study of interleukin-10, tumor necrosis factor-alpha, anti-U1-snRNP antibody levels and disease activity in patients with mixed connective tissue disease. AB - OBJECTIVE: To investigate the levels and relationship between IL-10, TNF-alpha, anti-U1snRNP antibodies and disease activity in longitudinally collected serum samples from patients with mixed connective tissue disease (MCTD). METHODS: Six patients followed for 17-138 months were investigated with ELISA for estimation of cytokine levels and antibodies to the different epitopes of the U1snRNP. Disease activity was assessed by systemic lupus activity measure (SLAM). RESULTS: IL-10 and TNF-alpha levels fluctuated with time in at least half of the patients. Three patients had increased IL-10 levels and two had increased TNF-alpha in all samples. There was no correlation between cytokine levels and disease activity or clinical manifestations. All patients had increased levels of antibodies to the main components of the U1snRNP. Both antibody levels and disease activity decreased with time. A correlation between TNF-alpha and U1snRNP antibody levels were observed in five patients. CONCLUSIONS: Increased and fluctuating levels of IL-10 or TNF-alpha without correlation to disease activity were observed in MCTD patients. In some patients increased cytokine levels were observed over several years irrespective of disease activity indicating that they could be constitutively increased in these individuals. PMID- 11727844 TI - Relative roles of endothelial cell damage and platelet activation in primary Raynaud's phenomenon (RP) and RP secondary to systemic sclerosis. AB - OBJECTIVE: To evaluate the relative roles of endothelium and platelets in the pathogenesis of primary RP and RP secondary to SSc. METHODS: Endothelial derived ET-1, t-PA, PAI-1, and platelet derived beta-TG, PDGF, TGF-beta were measured in 36 patients with primary RP, 14 patients with RP secondary to SSc and 30 age and sex matched controls. RESULTS: A significative increase of ET-1, t-PA, PAI-1, TGF beta, and beta-TG were the most relevant changes in patients with RP secondary to SSc with respect to the controls. Less relevant increases of t-PA, PAI-1, PDGF, and beta-TG levels were observed in patients with primary RP vs controls. CONCLUSIONS: These data seem to confirm the involvement of endothelial cells and platelets in the pathogenesis of RP, with mild changes in primary RP and more relevant changes in RP secondary to SSc. PMID- 11727845 TI - Serum YKL-40 concentrations in patients with early rheumatoid arthritis: relation to joint destruction. AB - OBJECTIVE: YKL-40 is a secretory glycoprotein of chondrocytes, synovial cells, macrophages, and neutrophils. The aims were to determine serum YKL-40 in patients with early rheumatoid arthritis (RA) and seek associations with early joint erosions. METHODS: YKL-40 was measured by ELISA in serum samples collected every three month for 36 months from patients with early RA. The patients were treated with DMARDs and some were allocated to additional prednisolone. RESULTS: Serum YKL-40 was higher in RA patients compared with controls (98 vs. 42 microg/l, p<0.001). The mean serum YKL-40 during the study correlated with the progression in Larsen score (Pearson's test: p=0.004). Patients with a persistently high serum YKL-40 had larger progression in Larsen score compared with patients with normal serum YKL-40 (median progression: 7 vs. 0, p=0.003). CONCLUSION: These data suggest that elevated serum YKL-40 is related to progression in joint destruction in early RA patients. PMID- 11727846 TI - Changes in plasma homocysteine in arthritis patients starting treatment with low dose methotrexate subsequently supplemented with folic acid. AB - In 15 patients with rheumatoid arthritis (RA) (n = 13) or psoriatic arthritis (PsA) (n = 2) p-homocystcine and erythrocyte folate (erc-FA) were measured before start of methotrexate (MTX) treatment, after 4 weeks of MTX treatment (median 10 mg per week), and after further 4 weeks of treatment with MTX (median 12.5 mg per week) supplemented with folic acid (FA) (15 mg per week). Mean p-homocysteine were 12.3 +/- 3.4 micromol/l, 14.6+/-5.8 micromol/l (p<0.05) and 10.3+/-3.0 micromol/l (p<0.01) respectively. P-homocysteine concentrations were negative correlated to erc-FA after 4 weeks (rho -0.58; p<0.05). It is concluded that treatment with MTX induces a significant rise in p-homocysteine that is neutralised by FA supplementation. Supplementation with FA from the start of MTX treatment is recommended considering the increased risk of cardiovascular disease that is associated with elevated concentrations of p-homocysteine. PMID- 11727847 TI - Thrombotic thrombocytopenic purpura and systemic lupus erythematosus. AB - Thrombotic thrombocytopenic purpura (TTP) is a rarely seen complicating systemic lupus erythematosus (SLE). The diagnosis of TTP in a setting of SLE is challenging since both share common features including thrombotic microangiopathy. We report two cases of SLE with TTP one with a good response when cyclophosphamide was given early with plasmapheresis and steroids; the other with a poor outcome in a patient given cyclophosphamide late in the course of the disease. PMID- 11727849 TI - Correcting the unfavorable outcomes following facelift surgery. AB - Although many deformities formerly observed after rhinoplasties are seen less because of more sophisticated techniques, the deformities that may occur after conventional facelifts seem to be growing by epidemic numbers because of the widespread and almost universal use of conventional facelift and blepharoplasty techniques. The unfortunate appearance of these patients is the fault neither of the surgeon nor of the patient, but frequently both get blamed. Throughout every step of the history of medicine there have been improvements made that cause the physician to abandon a technique or medications that clearly are shown to be suboptimal. Unfortunately, this has not happened with rejuvenative surgery. More sophisticated techniques like composite rhytidectomy have a steeper learning curve, take more time to perform, and require longer periods of convalescence for the patient. Many experts in aesthetic surgery continue to teach conventional techniques because the public does not seem to know the difference. Fashion magazine articles on aesthetic surgery promote simple and easy techniques without bothering to explain the unfortunate and minimal results that are obtained. The obligation of any surgeon doing aesthetic surgery is the same as any physician must have in any medical field. If any operation would give a patient a suboptimal or negative result, then attempts must be made to improve these techniques. Although traditional techniques will continue to be used, there are certain conclusions that can be made: 1. More people who have undergone plastic surgery are showing eventual signs of surgery that are neither attractive nor youthful. It is clear that results of conventional facelift techniques are not predictable and are often unfavorable. 2. Facelift deformities are not the fault of the surgeon but are natural byproducts of the techniques that are used. These techniques must be improved. 3. Composite rhytidectomy with orbital fat preservation as a primary procedure will create a harmonious facial rejuvenation that will disallow the appearance of unwanted deformities such as hollow eyes and the lateral sweep. 4. Composite rhytidectomy as a secondary procedure produces the most effective correction of the unintentional but unfavorable results that follow traditional rejuvenative surgery. PMID- 11727848 TI - Does mechanical insult to cartilage trigger relapsing polychondritis? PMID- 11727850 TI - Evolution of lower lid support following lower lid/midface rejuvenation: the pretarsal orbicularis lateral canthopexy. AB - An 18-month experience with an endoscopically assisted "closed" technique of modified transblepharoplasty lower lid and midface rejuvenation is presented. In the modified approach, preseptal dissection in the body of the lower lid is eliminated to decrease the incidence of septal scarring and lower lid retraction. The technique also leaves intact the connection between the orbicularis and septum, resulting in tightening of the septum and better support of lid with redraping of the muscle. By providing more secure support of the lower lid and lateral canthus, the need for direct canthal manipulation has been reduced significantly. This experience confirms, however, that an occasional patient with significant exophthalmus occasionally might need additional lower lid support (spacer graft or more direct lateral tendon fixation) to obtain ideal lower lid position. Conversely, an occasional older patient with relative enophthalmus and horizontal lower lid laxity will be best served by canthoplasty with correction of excess laxity. So far, the quality of aesthetic results compared with the "open" approach has not been compromised, and morbidity seems to have been decreased. PMID- 11727851 TI - The use of bioresorbable material in craniofacial surgery. AB - It is clear that in just the short period of time that these products have been available significant technical advances continue to occur to make these materials easier to employ and use. Technical advances will continue to occur to make them more adaptable and strong for all forms of use in the craniomaxillofacial skeleton. The clinician also can envision them being employed as stable carriers to allow other factors to be brought into a wound-healing environment, such as growth factors or bone morphogenetic proteins. In addition, because the concepts behind bioresorbable fixation can continue to be extrapolated, the elimination of plates and screws with replacement with appropriate glues can be imagined. As long as the concepts of biocompatibility and immunogenicity are adhered to, the future seems without limitations for bioresorbable fixation products, and the author would urge strongly that plastic and reconstructive surgeons remain in the forefront of this movement. PMID- 11727852 TI - New directions in the management of cleft lip. AB - Advances in cleft lip and palate have included improved understanding of its etiopathogenesis, genetics, and the surgical evolution of technical refinements in the correction of this deformity. PMID- 11727853 TI - Reconstructive microsurgery: reviewing the past, anticipating the future. AB - This article gives an overview of reconstructive microsurgery It is written for residents and fellows training in plastic and microsurgery and practitioners who are intimately involved in the clinical practice of complex reconstructions. The authors describe the highlights of the history of microsurgery and relevant clinical principles. The final portion of this article is designed to elucidate some of the tremendous advances in limb transplantation and the future directions of microsurgery. PMID- 11727854 TI - Future directions in breast implant surgery. AB - Breast implant surgery will remain a major component of the practice of plastic surgery. The future may hold promise for the introduction of new implants for the augmentation of the breast and for reconstruction. The constant demand and pressure placed on the manufacturers to keep developing new fillers are proof that satisfaction with saline implants is lacking. Today, however, the saline implant may be believed to be the safest breast implant available for clinical application. The demand necessitates that implants be used for enhancement of a woman's self image and for reconstruction. The smooth-surface implants are considered more popular and safer for patients, perhaps because it has been well documented that the shell may have been the cause of the problem. However, the constancy in holding on using and reapplying the same principles of a faulty shell may not be as scientific an endeavor as we have done in the past, and to keep placing different fillers in the faulty shell. Perhaps the lamination process used in manufacturing, the air in the shell, or wear and tear produces mechanical weakening of the shell that eventually will lead to loss of integrity of the silicone-shell breast implant. The global community is looking to plastic surgeons for a solution. The applications and the demand for breast implants are global in nature. Meanwhile, as clinicians are waiting for a new implantable breast device, they will continue to use what is available and advise patients that the implantation of a breast prosthesis is not a life-long endeavor. There is a need, however, for the implants to be maintained by the process of exchange every 8 to 10 years. Breast reaugmentation is to be performed as a standard because plastic surgeons will be waiting for further clarifications from the regulators or the scientific community. The goal is to produce a good outcome and maintain safety for the patients with a high standard of care. PMID- 11727855 TI - New perspectives in facial contouring using external ultrasonography. AB - The use of XUAL ultrasonic energy to "fractionate" and redistribute facial fat is a valuable adjunct in facial plastic surgery. The 59 patients enrolled in the author's preliminary evaluation of XUAL under the auspices of the American Society of Aesthetic Plastic Surgery Innovative Procedures Committee had in addition to the body liposculpture external ultrasonography application to "superwet" anesthetized face and neck with or without physical removal of fat or skin. When no excisions or liposuctioning were performed, there was visible and photographic improvement in facial contouring. Interview comments ranged from "I can see my cheek bones now" to questions as to whether or not a facelift had indeed been performed. Individuals who had submental resculpturing ("submental tuck") or simple "safe zone" liposuction in the submental area, jowl, and nasolabial zones also showed a remarkable degree of skin tightening and contouring beyond the area of actual fat removal. Those individuals who have been observed closely for more than 12 months still have the improvement. Redistribution of fat and skin tightening initially noted between the second and eighth weeks have persisted unchanged, often in spite of fat accumulation elsewhere from weight gain. PMID- 11727856 TI - New directions in plastic surgery research. AB - Plastic surgery research affords tremendous opportunities in a variety of affluent mode systems. Only recently have researchers applied molecular biologic techniques to common plastic surgery problems. For example, investigating the fundamental biomolecular mechanisms of normal palate and cranial suture morphogenesis will improve the understanding of the etiopathogenesis of CLP and craniosynostosis and facilitate the development of biologically-based interventions. Furthermore, as interdisciplinary collaborations improve, surgeons can expect to see remarkable progress in de novo tissue synthesis, replacement, and repair. Ultimately, they may one day find that gene-modified endogenous tissue engineering will succeed today's biocompatible scaffolds and allogeneic or zenogeneic replacement strategies. In general, plastic surgeons can look forward to the development of highly effective biomolecular treatments for clinical problems such as complex wound repair, prolific scarring, bone deficits (or surpluses), and organ system replacement or repair. Researchers believe that biologically-based strategies like these will be combined with technical advances that harness minimally invasive approaches. Together, clinicians expect these new tactics will reduce morbidity and improve the results of clinical problems treated by plastic surgeons. PMID- 11727857 TI - Pediatric plastic surgery revisited. AB - Improvements in surgical technique and an ever improving understanding of the embryologic and cellular origin of many congenital anomalies marks the gains made during the last decade in pediatric plastic surgery. This contemporary review highlights many of these advancements in the diverse array of birth and surgical defects. PMID- 11727858 TI - New directions in skin care. AB - Tretinoin and AHAs are dependable components of any skin-rejuvenation program. These products are well studied, and the results are predictable. Antioxidants make biologic sense; the initial claims were far-reaching, but there is evidence to support their use in the prevention of photoaging. Microdermabrasion has many potential applications. Epidermal level treatment is enjoying popular appeal; however, the efficacy for epidermal level treatment of photoaging has yet to be established. New products and treatments will come, and others will go. Most importantly, clinicians should remain scientists, not salespeople, in the quest for the silver bullet to combat photoaging. PMID- 11727859 TI - Polycystic ovary syndrome and cancer. AB - The polycystic ovary syndrome (PCOS) is the most common endocrine disturbance affecting women, but disagreements in diagnostic criteria make it difficult to compare epidemiological studies on long-term health risks such as cancer. The association between PCOS and endometrial adenocarcinoma has been reported for many years. Although the degree of risk has not been clearly defined, it is generally accepted that for women with PCOS who experience symptoms of amenorrhoea or oligomenorrhoea, the induction of artificial withdrawal bleeds to prevent endometrial hyperplasia is prudent management. Studies examining the relationship between PCOS and breast carcinoma have not always identified a significantly increased risk, although one recent study examined the standardized mortality rate (SMR) calculated for patients with PCOS compared with the normal population and found that the SMR for all neoplasms was 0.91 (95% CI 0.60-1.32) and for breast cancer 1.48 (95% CI 0.79-2.54). Few studies have addressed the possibility of an association between polycystic ovaries and ovarian cancer, and the results are conflicting and generally reassuring. PMID- 11727860 TI - Ovarian tissue cryopreservation and transplantation: preliminary findings and implications for cancer patients. AB - Cancer treatment modalities are increasingly more effective in achieving complete remission and cure. Aggressive chemotherapy and radiotherapy, as well as bone marrow transplantation, results in >90% cure in many cancers of children and young women. As a result of this success however, a new problem has arisen. Many young women survive to live the rest of their lives in menopause, and have no chance of conceiving on their own. Oocyte cryopreservation has resulted in a handful of pregnancies, but the technique may not be applicable to young women and children. Ovarian tissue cryopreservation and transplantation has emerged against this background, first in successful rodent studies, and then in sheep and human ovarian xenograft studies. Because of the encouraging results with animals and xenografts, a human ovarian transplantation trial was launched. Pelvic auto-transplantation of frozen-banked ovarian tissue resulted in ovulation in one patient. Several other patients received fresh grafts subcutaneously, and preliminary results indicated antral follicle development at least in one patient. With the addition of promising data from humans, ovarian tissue cryopreservation from selected patients before cancer treatment, and in those requiring oophorectomy for benign causes, is now advocated. PMID- 11727861 TI - The effects of radiotherapy and chemotherapy on female reproduction. AB - High dose chemotherapy and radiotherapy have radically increased long-term survival of young cancer patients, but major side effects of these treatments are ovarian failure and infertility. Knowledge of the risks and probabilities of ovarian failure caused by treatment is crucial for patients and physicians in order to make informed choices that will best serve patients' interests. This review presents data on ovarian damage and failure following exposure to radiotherapy, chemotherapy and ablative therapy. The risk is evaluated from the published literature according to patient's age, treatment protocol and also according to the diagnosis of some common malignancies. Many of these patients will not be sterilized immediately following treatment, but will suffer from premature menopause. In order to prevent sterilization, ovarian transposition before pelvic irradiation is mandatory. Besides cryopreservation of ovarian tissue and embryos before administration of chemotherapy, the possible protective effect of pituitary-ovarian down-regulation is discussed. The mechanism of primordial follicles damage induced by radio/chemotherapy is presented as well as the role of apoptosis signalling pathways underlying destruction. Increased knowledge of these mechanisms could help to identify potential effective inhibitors that can block the path of primordial follicles destruction and reduce ovarian failure rate. PMID- 11727862 TI - Introduction--prevention of adhesion formation: the journey continues. AB - One of the complications of abdominal surgery is the occurrence of intra abdominal adhesions. The problems related to these adhesions have created a major impact on the health care system, thereby incurring a substantial cost to health care. In the field of gynaecology, adhesion-related problems include infertility, abdominal pain and bowel obstruction. PMID- 11727863 TI - Peritoneal repair and post-surgical adhesion formation. AB - It was shown in 1919 that peritoneal healing differs from that of skin. When a defect is made in the parietal peritoneum the entire surface becomes epithelialized simultaneously and not gradually from the borders as in epidermalization of skin wounds. While multiplication and migration of mesothelial cells from the margin of the wound may play a small part in the regenerative process, it cannot play a major role, since new mesothelium develops in the centre of a large wound at the same time as it develops in the centre of a smaller one. Development of intraperitoneal adhesions is a dynamic process whereby surgically traumatized tissues in apposition bind through fibrin bridges which become organized by wound repair cells, often supporting a rich vascular supply as well as neuronal elements. PMID- 11727864 TI - Peritoneal healing and adhesion formation/reformation. AB - Intra-abdominal adhesion formation and reformation after surgery is a cause of significant morbidity, resulting in infertility and pain. The understanding of the pathogenesis of adhesion formation and reformation especially at the cellular and molecular level can help to further develop more effective treatments for the prevention of adhesion formation and reformation. Following an injury to the peritoneum, fibrinolytic activity over the peritoneal surface decreases, leading to changes in the expression and synthesis of various cellular mediators and in the remodelling of the connective tissue. The cellular response to peritoneal injury and adhesion formation and reformation are reviewed. Analysis of the available literature data on the cellular mediators in the peritoneal fluid showed variation in results from different investigators. The potential sources of variability and error are examined. It is still unclear if there is significant individual variation in the peritoneal response to injury. PMID- 11727865 TI - Clinical implications of postsurgical adhesions. AB - Adhesion development can have a major impact on a patient's subsequent health. Adhesions are a significant source of impaired organ functioning, decreased fertility, bowel obstruction, difficult re-operation, and possibly pain. Consequently, their financial sequelae are also extraordinary, with more than one billion dollars spent in the USA in 1994 on the bowel obstruction component alone. Performing adhesiolysis for pain relief appears efficacious in certain subsets of women. Unfortunately even when lysed, adhesions have a great propensity to reform. Adhesions are prevalent in all surgical fields, and nearly any compartment of the body. For treatment of infertility and recurrent pregnancy loss, lysis of intrauterine adhesions results in improved fecundability and decreased pregnancy loss. PMID- 11727866 TI - Evidence-based prevention of post-operative adhesions. AB - Despite decades of research, numerous new product ideas and 'carefully considered opinions' of recognized experts, very few products for the prevention of post operative adhesions have met the requirements for Level 1 evidence of safety and efficacy. Those that have are useable only at laparotomy. Several new liquid products intended for use at laparoscopy are in various stages of development and clinical investigation. Hopefully, some will prove to be both simple to use and efficacious. Even if this occurs, it must be remembered that a reduction in post operative adhesions does not necessarily produce a better clinical outcome. Our common sense suggests that fewer adhesions should logically result in less pain, more pregnancies, fewer bowel obstructions and less long-term morbidity. We believe that 'fewer adhesions' is a good thing, but we have no controlled human trials to prove this. How much of a reduction in post-operative adhesions is necessary before it is clinically relevant? A single adhesion in the wrong anatomic location may be catastrophic. How do we measure this? Until these and other questions have been answered (if ever), we have nothing more than educated guesses that all these efforts are warranted. PMID- 11727867 TI - Ovarian reserve testing and the use of prognostic models in patients with subfertility. AB - The decline in fecundity with female age is a well-known phenomenon for clinicians dealing with subfertility patients. Diminishing ovarian reserve seems to be the reason for declining fecundity. Since age is only a rough estimate of ovarian reserve, many tests have been developed to predict ovarian reserve more precisely. This review focuses on these ovarian reserve tests and their clinical role in predicting response to ovarian stimulation and pregnancy chances. According to our analysis, the clomiphene citrate challenge test has the strongest correlation in predicting ovarian reserve, and is the only test that is validated in the general infertility population. The antral follicle count by ultrasound is promising and may offer clinical use. It is not known whether a combination of tests can provide more accurate information of ovarian reserve. It is not yet clear to which extent the results of ovarian reserve tests can be incorporated into the available prognostic models. There is a need for prospective cohort studies that focus on prognostic factors among which are the results of ovarian reserve tests. Only then can the qualitative and quantitative relevance of ovarian reserve testing in the context of the prognosis for couples with subfertility be established. PMID- 11727868 TI - PGD in female carriers of balanced Robertsonian and reciprocal translocations by first polar body analysis. AB - Preimplantation genetic diagnosis (PGD) using the first polar body (1PB) is a modality of PGD that can be used when the woman is the carrier of a genetic disease or of a balanced chromosomal reorganization. PGD using 1PB biopsy in carriers of balanced chromosome reorganizations has not become generalized. Here, we describe our experience based on the analysis of unfertilized or fresh, non inseminated control oocytes, by fixing separately the 1PB and the corresponding oocyte, and on the study of six clinical cases of PGD using 1PB biopsy (four Robertsonian translocations and two reciprocal translocations). In fresh oocytes, the chromosome morphology of the 1PB was well preserved, and the results were always concordant for each oocyte-1PB pair. This indicates that the 1PB can be reliably used for the diagnosis of chromosome reorganizations. In these studies the technical problems encountered when performing PGD using 1PB biopsies for chromosome studies are also addressed. Three different strategies of 1PB biopsy (laser beam, partial zona dissection and acid Tyrode's) and two different protocols (intracytoplasmic sperm injection before or after 1PB biopsy) and their effect on the percentage of oocytes diagnosed and the fertilization rate, are discussed. In reciprocal translocation cases, published in the literature or studied by us, in which at least nine oocytes had been diagnosed, a correlation has been found between the frequency of nondisjunction observed and the theoretical recombination rate. To date, PGD by 1PB analysis alone or combined with blastomere biopsies in female carriers of chromosomal rearrangements has been used in 18 cases, with a further six cases reported here. A total of 325 cumulus-oocyte complexes have been obtained, of which 294 were biopsied and 224 were diagnosed. A total of 52 embryos was transferred, 19 of which implanted and 17 produced full-term pregnancies. PMID- 11727870 TI - Some afterthoughts about eclipse blindness. PMID- 11727869 TI - Turner's syndrome and fertility: current status and possible putative prospects. AB - Women with Turner's syndrome should be carefully followed throughout life. Growth hormone therapy should be started at age 2-5 years. Hormone replacement therapy for the development of normal female sexual characteristics should be started at age 12-15 years and continued for the long term to prevent coronary artery disease and osteoporosis. Most women with Turner's syndrome have ovarian dysgenesis; therefore, they are usually infertile, and in very rare cases have spontaneous menses followed by early menopause. Only 2% of the women have natural pregnancies, with high rates of miscarriages, stillbirths and malformed babies. Their pregnancy rate in oocyte donation programmes is 24-47%, but even these pregnancies have a high rate of miscarriage, probably due to uterine factors. A possible future prospect is cryopreservation of ovarian tissue containing immature follicles before the onset of early menopause, but methods of replantation and in-vitro maturation still need to be developed. Should these autologous oocytes indeed be used in the future, affected women would need to undergo genetic counselling before conception, followed by prenatal assessment. PMID- 11727871 TI - Ocular signs of systemic hypertension: a review. AB - Cardiovascular disease and stroke continue to be the chief causes of death in developed countries and one of the leading causes of visual impairment. The individual with systemic hypertension may remain asymptomatic for many years. Systemic mortality and morbidity are markedly higher for hypertensives than normotensives, but can be significantly reduced by early diagnosis and then efficient management. However, the ability of Optometrists to detect and appropriately refer systemic hypertensives remains generally poor. This review examines the disease, its effects and detection by observation of the retinal signs, particularly those considered to be pre-malignant. Previous methods of classifying retinal hypertensive signs are discussed along with more recent image analysis techniques. The role of the optometrist in detecting, monitoring and appropriate referral of systemic hypertensives is discussed in relation to current research. PMID- 11727872 TI - Improving the description of the retinal vasculature and patient history taking for monitoring systemic hypertension. AB - Systemic hypertension is an important public health concern. If optometrists are to perform a more active role in the detection and monitoring of high blood pressure (BP), there is a need to improve the consistency of describing the retinal vasculature and to assess patient's ability to correctly report the diagnosis of hypertension, its control and medication. One hundred and one patients aged > 40 years were dilated and had fundus photography performed. BP was measured and a self-reported history of general health and current medication was compared with the records of their general practitioner (GP). The status of the retinal vasculature was quantified using a numeric scale by five clinicians and this was compared to the same evaluation performed with the aid of a basic pictorial grading scale. Image analysis was used to objectively measure the artery-to-vein (A/V) ratio and arterial reflex. Arteriolar tortuosity and calibre changes were found to be the most sensitive retinal signs of high BP. Using the grading scale to describe the retinal vasculature significantly improved inter- and intra-observer repeatability. Almost half the patients examined were on medication for high BP or cardiovascular disease. Patients' ability to give their complete medical history was poor, as was their ability to recall what medication they had been prescribed. GPs indicated it was useful to receive details of their patient's BP when it was > 140/90 mmHg. The use of improved description of the retinal vasculature and stronger links between optometrists and GPs may enhance future patient care. PMID- 11727873 TI - Phakometric measurement of ocular surface radius of curvature and alignment: evaluation of method with physical model eyes. AB - Ophthalmophakometric measurements of ocular surface radius of curvature and alignment were evaluated on physical model eyes encompassing a wide range of human ocular dimensions. The results indicated that defocus errors arising from imperfections in the ophthalmophakometer camera telecentricity and light source collimation were smaller than experimental errors. Reasonable estimates emerged for anterior lens surface radius of curvature (accuracy: 0.02-0.10 mm; precision 0.05-0.09 mm), posterior lens surface radius of curvature (accuracy: 0.10-0.55 mm; precision 0.06-0.20 mm), eye rotation (accuracy: 0.00-0.32 degrees; precision 0.06-0.25 degrees), lens tilt (accuracy: 0.00-0.33 degrees; precision 0.05-0.98 degrees) and lens decentration (accuracy: 0.00-0.07 mm; precision 0.00-0.07 mm). PMID- 11727874 TI - Diagnosis of eccentric fixation using a calibrated ophthalmoscope: defining clinically significant limits. AB - Visuscopy and other ophthalmoscopic methods are widely applied in the assessment of fixation behaviour in amblyopia, although the reliability and inter-examiner variability of this test has not been established. Typically eccentric fixation is diagnosed solely on the basis of retinal position, but this fails to address the accompanying sensorimotor adaptations. A double-blind (masked) trial of a paediatric population was undertaken (n = 30, age range 3 years 9 months to 11 years, mean 5.6 +/- 1.5 years), involving three examiners applying a detailed protocol. The criteria for eccentric fixation were established from the non amblyopic eyes. The most reliable criterion for the diagnosis of fixation was established from the results of the study. No single parameter proved 100% reliable, and amplitude showed greatest inconsistencies. It is recommended that a consensus of at least three parameters from position, zero retinomotor point, principal visual direction and percent foveation is required to diagnose the fixation status reliably. PMID- 11727875 TI - The effects of accommodative facility training on a group of children with impaired relative accommodation--a comparison between dioptric treatment and sham treatment. AB - The effects of accommodative facility training were evaluated by comparing training with plano lenses and +/-2.00 D lens flipper sets. Thirteen children with symptoms and signs of accommodative dysfunction were included in the study. Seven started with 2 weeks of plano lens training before proceeding to traditional dioptric flipper training; later, one patient from this group was lost to follow-up. The remaining six children used powered flipper training from the start. The positive (PRA) and negative relative accommodation (NRA) were examined every second week. During the initial training period both the PRA and the NRA decreased in the sham treatment group (P = 0.010 and P= 0.102, respectively), while the PRA and the NRA increased during the dioptric training in both groups (P = 0.102 and P = 0.033, respectively). The result of this study indicates that accommodative facility training has a real effect on the amplitude of relative accommodation in patients with impaired relative accommodation. PMID- 11727876 TI - The repeatability and accuracy of axial length and anterior chamber depth measurements from the IOLMaster . AB - BACKGROUND: Axial length and anterior chamber depth have been measured clinically using conventional ultrasound biometry. Recently, a non-contact device has become available to measure these parameters. This study evaluated the repeatability and accuracy of this device. METHODS: The axial length and anterior chamber depth were measured by two practitioners on a group of young subjects using the IOLMaster followed by a conventional ultrasound biometer operated by a third practitioner. The accommodation was controlled in ultrasound biometry through a full correction on the non-measured eye and a distant fixation target. RESULTS: There was good repeatability and accuracy of axial length assessment. The mean difference between the IOLMaster and ultrasound biometry was -0.099 mm, with 95% limits of agreement between 0.66 and -0.85 mm. The axial length was slightly shorter from the IOLMaster and the difference was not significant. The anterior chamber depth was repeatable but was shown to be deeper than the ultrasound results. The mean difference in anterior chamber depth between the IOLMaster and ultrasound biometry was 0.15 mm, with 95% limits of agreement between 0.34 and 0.03 mm. It is suggested that the former device is not measuring the axial anterior chamber depth. CONCLUSIONS: The IOLMaster is a non-contact 'optical' A scan which is simple to use and good for axial length assessment. The anterior chamber depth assessment should be further evaluated. PMID- 11727877 TI - The relation between accommodative facility and general binocular dysfunction. PMID- 11727878 TI - Winners in the 2000 Naylor Prize Competition. PMID- 11727879 TI - Preadolescent predictors of substance initiation: a test of both the direct and mediated effect of family social control factors on deviant peer associations and substance initiation. AB - A review of the literature suggests that early substance initiation is related to a variety of negative outcomes, including substance misuse or abuse in adolescence and adulthood. This study examines potentially modifiable predictors of early substance initiation, including both family and peer factors known to influence early initiation. A theoretically derived model of substance initiation was tested using structural equation modeling. Results indicate that both family and peer factors have an impact on early substance initiation when children in this sample were 11 and 12 years old. The model explained 60% of the variance in substance initiation. Prosocial family processes (rules, monitoring, and attachment) had a significant impact on child peer association, decreasing involvement with antisocial peers. These prosocial family processes had a significant negative effect on substance initiation even while modeling the influence of antisocial peers. Implications for drug use prevention practice are discussed. PMID- 11727880 TI - Counseling frequency and the effectiveness of outpatient drug treatment: revisiting the conclusion that "more is better.". AB - Replicating and extending previous research, findings from a prospective investigation of outpatient drug treatment (n = 356) indicate that higher frequency of participation in group counseling during treatment predicts higher rates of alcohol and drug abstinence even for those who complete the 6-month treatment program and maintain weekly or more frequent attendance of 12-step meetings during and after treatment. Greater length of group and individual counseling sessions, however, is not associated with higher rates of abstinence, and contrary to previous research, the modest but statistically significant association between the frequency of individual counseling and abstinence was not replicated. With respect to treatment completion, regular attendance at 12- step meetings, and frequent participation in group counseling while in treatment, the evidence suggests that "more is better." PMID- 11727881 TI - Treatment process in prison therapeutic communities: motivation, participation, and outcome. AB - Although the largest effects of prison-based therapeutic community (TC) programs are associated with entry into aftercare, only a minority of prisoners volunteer for these aftercare programs. The study addresses the gap in our knowledge concerning these low rates of voluntary entry. A theoretical formulation of the TC process involving the effect of the interaction of clients' motivation and participation in the activities of the TC on entry into aftercare was tested on a sample of 110 volunteers in a prison-based TC for whom there were client and staff ratings of 3-month participation and 12-month follow-up data on relapse and recidivism. Path analyses support a model in which the interaction of motivation and 3-month participation ratings have a direct effect on the selection of aftercare, and aftercare has a direct effect on relapse and recidivism. The use of a combination of enhanced motivation and early program participation as a means of increasing the utilization and effectiveness of aftercare is discussed. PMID- 11727882 TI - Multidimensional family therapy for adolescent drug abuse: results of a randomized clinical trial. AB - Random assignment was made of 182 clinically referred marijuana- and alcohol abusing adolescents to one of three treatments: multidimensional family therapy (MDFT), adolescent group therapy (AGT), and multifamily educational intervention (MEI). Each treatment represented a different theory base and treatment format. All treatments were based on a manual and were delivered on a once-a-week outpatient basis. The therapists were experienced community clinicians trained to model-specific competence prior to the study and then supervised throughout the clinical trial. A theory-based multimodal assessment strategy measured symptom changes and prosocial functioning at intake, termination, and 6 and 12 months following termination. Results indicate improvement among youths in all three treatments, with MDFT showing superior improvement overall. MDFT participants also demonstrated change at the 1-year follow-up period in the important prosocial factors of school/academic performance and family functioning as measured by behavioral ratings. Results support the efficacy of MDFT, a relatively short-term, multicomponent, multitarget, family-based intervention in significantly reducing adolescent drug abuse and facilitating adaptive and protective developmental processes. PMID- 11727883 TI - Deaths related to the inhalation of volatile substances in Texas: 1988-1998. AB - Analysis of death certificates in Texas from 1988 to 1998 showed that the characteristics of 144 Texans for whom inhalants were mentioned as a contributing cause of death are different from those Texans who report use of inhalants in surveys and from persons who died from inhalant abuse in Virginia. While Texas surveys show little difference in prevalence of use between white and Hispanic adolescents or between boys and girls, Texas death data indicate inhalant use is also a problem among adult white males. The mean age of decedents was 25.6 years (SD 9.8 years), and ages ranged from 8 to 62 years. There were 92% males, 81% whites, and 17% Hispanics. Of the death certificates, 35% mentioned Freon, and 25% mentioned chlorinated hydrocarbons. Of those with the mention of Freon, 42% were students (mean age 16.4 years), and 37% were mechanics, installers, and repairers (mean age 27.4 years), occupations in which Freon can be readily available. Of the chlorinated hydrocarbon deaths, 49% were students (mean age 17.5 years), and 51% were from other occupations (mean age 27.4 years). Research on drug use and the workplace is not extensive, and the effects of inhalant intoxication can be a serious problem in the workplace. Prevention campaigns need to inform the public that inhalant abuse is not just a problem among youngsters, and intervention services for adult abusers should be provided within the context of employee assistance programs. PMID- 11727884 TI - Diagnosing antisocial personality disorder among substance abusers: the scid versus the MCMI-II. AB - There is much controversy among social scientists and clinicians over the proper measurement of antisocial personality disorder (ASPD). The degree to which various diagnostic measures differ in their assessment of ASPD among substance abusers is not known. This study assessed the degree of agreement between a semistructured clinical interview and a self-report inventory on a diagnosis of ASPD among substance abusers. The Structured Clinical Interview for DSM-III-R (SCID-II), a clinically generated instrument, and the Millon Clinical Multiaxial Inventory (MCMI-II), a self-report inventory, were administered to 275 clients randomly assigned to two therapeutic communities (TCs). Based on the limited existing literature, it was hypothesized that there would be minimal agreement between the diagnosis of ASPD by the two scales. This hypothesis was supported. The kappa statistic indicated low agreement between the scales (kappa = 0.27), with the MCMI-II diagnosing ASPD more often than the SCID-II. The low agreement on a diagnosis of ASPD may be due to the different types of information collected by the two scales. The SCID-II emphasizes observable behavioral criteria, while the MCMI-II emphasizes pathological personality traits. The focus of the MCMI-II on pathological personality traits may more accurately diagnose ASPD in substance abusing populations in which the majority of the clients have extensive criminal histories. Definite conclusions regarding the proper measurement of ASPD in substance-abusing samples is difficult without additional empirical evidence. PMID- 11727885 TI - Residential status and HIV risk behaviors among Puerto Rican drug injectors in New York and Puerto Rico. AB - This article investigates the association between residential status and human immunodeficiency virus (HIV) risk behaviors among island and New York Puerto Rican injection drug users (IDUs). We assigned 561 subjects from New York City and 312 from Puerto Rico to five residential status categories: living in parent's home, living in own home, living in other's home, living in temporary housing (hotel, single-room occupancy [SRO] hotels), and homeless (living in streets/shelters). Dependent variables included injection- and sex-related risk behaviors (sharing syringes, sharing other injection paraphernalia, shooting gallery use, and having paid sex). Chi square, t tests, and multivariate logistic analysis tests were performed separately by site. About one-quarter of the sample in each site was homeless. Island Puerto Ricans were more likely to live with their parents (44% vs. 12%, p < .001), and more New York IDUs lived in their own home (30% vs. 14%, p < .001). In New York, gallery use and paid sex were associated with living in other's home, living in parent's home, and being homeless. Sharing paraphernalia was related to living in other's home, living in temporary housing, and being homeless. In Puerto Rico, having paid sex was associated with homelessness. High-risk behaviors were more likely among homeless IDUs in both sites. Programs to provide housing and target outreach and other prevention programs for homeless IDUs would be helpful in reducing HIV risk. PMID- 11727886 TI - The relationship between alcohol use and mortality rates from injuries: a comparison of measures. AB - Per capita consumption of alcohol has traditionally been considered to be the leading indicator of population levels of alcohol problems. However, some recent research suggests that this relationship may be weakening, and that drinking pattern measures may be preferable to per capita consumption as problem-level indicators. We compared the ability of per capita alcohol consumption and survey based measures of alcohol use to predict deaths from injuries in Ontario, Canada, for the period 1977-1996. Per capita consumption and percentage of daily drinkers were significantly related to injury mortality, but percentage of drinkers and percentage of episodic heavy drinkers (those who drank five or more drinks on a drinking occasion) were not. Of the measures we examined, per capita consumption was the strongest indicator of mortality rates from injuries. However, the survey derived measure of percentage of daily drinkers was similar to per capita consumption in ability to predict problem levels. PMID- 11727887 TI - A measure of the intensity of response to alcohol in a military population. AB - Heavy drinking and associated problems are relatively common in young men, including those in a military setting. This article explores characteristics of alcohol intake and associated difficulties and their relationship to a self report of the usual intensity of response to alcohol in a sample of U.S. Marines. Two questionnaires related to demography and alcohol use histories, along with a simple, 12-item self-report measure of the usual number of drinks to experience an effect (the Self-Rating of the Effects of Alcohol, SRE) were administered to 1320 U.S. Marines. The sample had an average age of 22 years, 78% were Caucasian, and 92% were enlisted personnel. The relationships and correlations among drinking characteristics and problems and the usual number of drinks for an effect were determined. These subjects drank an average of 6 days per month, consuming an average of almost six drinks per drinking day, and reported more than three times per month in which they consumed six or more drinks per occasion. Consistent with studies of other populations, the SRE measures of intensity of response to alcohol showed a positive correlation with both drinking practices and problems, with the latter remaining significant even after controlling for recent drinking practices. The prodigious level of alcohol intake and associated problems, along with the SRE scores, indicate that the Marine Corps personnel are at especially high risk for alcohol-related life problems. These data also support the potential usefulness of the SRE both in identifying individuals likely to have more severe alcohol profiles and in educating individuals regarding their levels of risk for alcohol abuse and dependence. PMID- 11727888 TI - Alcohol use, alcohol disorders, and the use of health services: results from a population survey. AB - Data from a population survey were used to explore relationships among drinking levels/patterns, alcohol dependence or abuse, and the use of emergency services, hospital admissions, and frequent visits to general practitioners in the past year. For both males and females, self-reported hospital admissions were less common among daily moderate drinkers than among lifetime abstainers. Among males, drinkers with no history of alcohol dependence or abuse were less likely to report being in hospital in the last year than lifetime abstainers. For females, some groups defined by drinking patterns/levels and current drinkers without symptoms of alcohol abuse or dependence were more likely to report using emergency services than lifetime abstainers. PMID- 11727889 TI - Alcohol use in high school: predicting students' alcohol use and alcohol problems in four-year colleges. AB - This research examined the effect of students' drinking behaviors in high school on their alcohol use and alcohol problems in college. Five colleges in New York State were randomly selected for the study. The telephone interview method was employed to survey college students during the spring semester of 1998. Within each participating college, students were randomly selected through the use of complete student telephone directories provided by the college administration. A total of 813 students were interviewed. Alcohol problems were measured with items from DSM-IV. Results indicate that both frequency and quantity of alcohol use in high school significantly affect students' alcohol consumption in college. Furthermore, the impact of alcohol use in high school on alcohol problems in college tends to be composed of a moderate direct effect and a relatively strong indirect effect via its impact on current alcohol consumption in college. Prevention efforts for drinking in college may need to start in high school to reduce or delay alcohol use by high school students. Additional assessment services and more readily available treatment services may be required to address potential alcohol problems among college students. Future research should examine the effect and longevity of students' alcohol problem symptoms after they graduate from college. PMID- 11727890 TI - Toward a new classification of risk factors for suicide behavior. PMID- 11727891 TI - Reaching out to survivors of suicide. PMID- 11727892 TI - Nonfatal suicidal behavior as a communication. PMID- 11727893 TI - Letter across the Pacific. PMID- 11727894 TI - Serotonergic genes and suicidality. AB - Several lines of evidence indicate that abnormalities in the functioning of the central serotonergic system are involved in the pathogenesis of depressive illness and suicidal behavior. Studies have shown that the number of brain and platelet serotonin transporter binding sites are reduced in patients with depression and in suicide victims, and that the density of 5-HT2A receptors is increased in brain regions of depressed in suicide victims and in platelets of depressed suicidal patients. Genes that code for proteins, such as tryptophan hydroxylase, 5-HT transporter, and 5-HT2A receptor, involved in regulating serotonergic neurotransmission, have thus been major candidate genes for association studies of suicide and suicidal behavior. Recent studies by our group and by others have shown that genetic variations in the serotonin-system-related genes might be associated with suicidal ideation and completed suicide. We have shown that the 102 C allele in 5-HT2A receptor gene was significantly associated with suicidal ideation (chi2 = 8.5. p < .005) in depressed patients. Patients with a 102 C/C genotype had a significantly higher mean HAMD item #3 score (indication of suicidal ideation) than T/C or T/7 genotype patients. Our results suggest that the 102T/C polymorphism in 5-HT2A receptor gene is primarily associated with suicidal ideation in patients with major depression and not with depression itself. We also found that the 5-HT transporter gene S/L polymorphism was significantly associated with completed suicide. The frequency of the L/L genotype in depressed suicide victims was almost double of that found in control group (48.6% vs. 26.2%). The odds ratio for the L allele was 2.1 (95% CI 1.2 3.7). The association between polymorphism in serotonergic genes and suicidality supports the hypothesis that genetic factors can modulate suicide risk by influencing serotonergic activity. PMID- 11727895 TI - What can psychiatric genetics offer suicidology? AB - There is good evidence from recent studies that depression is familial, and that a substantial proportion of the variation in liability is explained by genes. Suicidal behavior, including completed suicide, also seems to cluster in families. First-degree relatives of individuals who have committed suicide (included dizygotic twins) have more than twice the risk of the general population. For identical co-twins of suicides, the relative risk increases to about 11. Applying a simple structural equation model to the published data suggests a heritability for completed suicide of about 43% (95% confidence intervals 25-60). It is not known at present whether the genes predisposing to suicide are identical with those predisposing to affective disorder, but since only about half of those committing suicide have a diagnosis of depression, it seems probable that the overlap is incomplete. The mode of inheritance of suicidal behavior is almost certain to be complex, involving many genes. There have already been some initial studies of allelic association with polymorphisms in candidate genes such as those involved in serotonergic transmission. Further progress is likely to come from candidate gene and linkage disequilibrium studies that are capable of detecting multiple genes of small effect. PMID- 11727896 TI - Suicide, serotonin, and the brain. AB - The involvement of impaired serotonergic functioning in the development of suicidal behavior is one of the best documented findings in biological psychiatry. It is, however, less clear in which way this dysfunction contributes to the occurrence of suicidal behavior. Correlational studies have demonstrated associations between peripheral measures of serotonergic function and characteristics such as impulsivity, disinhibition, anxiety, and/or behavioral inhibition. Postmortem and neuroimaging studies have provided insight in the localization of serotonergic dysfunction in the central nervous system. Nevertheless, results in this area of research have also been contradictory. Following a short overview of recent research findings on serotonin and suicidal behavior, this paper focuses on the involvement of the prefrontal cortex of the brain in the development of suicidal behavior and on the role of serotonin in its executive functions. Based on these considerations, suggestions for future research are discussed. PMID- 11727897 TI - The Cardiff sib-pair study: suicidal ideation in depressed and healthy subjects and their siblings. AB - Depression is associated with high rates of suicidal ideation, which varies in intensity from transient thoughts of wishing to be dead to the making of plans and, Finally, to attempts to kill oneself. There is limited evidence from family, twin, and adoption studies that completed suicide is familial and has a genetic etiological component. However, it is unclear whether suicidal ideation is also familial. The familiality of suicidal ideation has been examined in the subjects who participated in the Cardiff Depression Study, namely, 108 depressed probands. their nearest-aged siblings, and 105 healthy control subjects and their siblings. The study showed that 66% of depressed subjects had experienced suicidal ideation in the week prior to the interview, and that this was significantly associated with recurrent illness. Suicidal ideation was not shown to be familial. However, somewhat surprisingly, 6% of healthy, never-depressed subjects admitted to having had transient suicidal thoughts. Suicidal ideation was significantly associated with high neuroticism and psychoticism scores and severe threatening life events. PMID- 11727898 TI - Can professionals and nonprofessionals work together following a suicide? AB - Suicide is the taking of one's own life by one's own hand. It is often sudden and creates many emotional reactions for the survivors left behind. Survivor responses can be impacted by a range of circumstances, from how the person died to the reactions of people to the survivor. This contribution examines the uniqueness of survivor grief and how best to help survivors. It is suggested that their grief may be too quickly viewed as pathological, resulting in the premature medicalization of basically normal reactions. It is time for professionals and nonprofessionals to recognize boundaries and work toward mutual goals of health for those persons left behind after a suicide. PMID- 11727899 TI - International meeting "Suicide: interplay of genes and environment". PMID- 11727900 TI - "Paediatric Cardiology Hall of Fame". PMID- 11727901 TI - How do we determine atrial arrangement? PMID- 11727902 TI - Neonatal complete correction of tetralogy of Fallot versus shunting and deferred repair: is the future of the right ventriculo-arterial junction at stake, and what of it? PMID- 11727903 TI - Hemorheology and renal function during cardiopulmonary bypass in infants. AB - BACKGROUND: Acute renal failure is an occasional complication after cardiopulmonary bypass in infants. Whereas it is well known that postoperative hemodynamics inflict acute renal failure, the influence of extracorporeal circulation on the kidney is less clear. Moreover, changes in blood viscosity occur during and after surgery, which may influence renal dysfunction. For this reason, we investigated the impact of blood viscosity on renal function during cardiopulmonary bypass. METHODS: In 34 patients weighting less than 10 kg, we performed repeated analysis of urine, blood, and plasma viscosity. RESULTS: Polyuria and proteinuria that appeared during cardiopulmonary bypass indicated an elevated transglomerular filtration gradient, which recovered within 24 hours. The appearance of N-acetyl-beta-D-glucosaminidase in the urine, and elevated excretion of sodium, were additionally indicative of mild tubular damage. Elevation of blood viscosity during hypothermic perfusion showed a statistical correlation with proteinuria and N-acetyl-beta-D-glucosaminidaseuria. With hypothermia, the relation of blood viscosity to plasma viscosity became stronger, while the relation to the hematocrit decreased compared to normothermia. CONCLUSIONS: During cardiopulmonary bypass perfusion, the kidney can be stressed by proteinuria and mild tubular damage. Our data provide evidence that the kidneys can be protected by improved blood viscosity during cardioplegia, but this needs confirmation in a prospective interventional study. PMID- 11727904 TI - The pathogenesis of heart failure in infants with congenital heart disease. AB - BACKGROUND: The clinical symptoms of heart failure in infants with left-to-right shunts are thought to be explained by well-known hemodynamic disturbances such as pulmonary hypertension and overcirculation, but previous studies have not, thus far, found the expected correlations with hemodynamic and clinical parameters. Based on the neurohormonal model of heart failure, we hypothesised that the clinical symptoms of infants with left-to-right shunts are also related to neurohormonal disorders. METHODS: We compared various neurohormonal and hemodynamic parameters measured invasively in 70 infants with left-to-right shunts to the respiratory rate and gain in weight over a corresponding period of time. Heart rate correlated significantly with respiratory rate (r = 0.62***, p < 0.001) and gain in weight (r = -0.31*, p = 0.015), but more conventional measures of severity, such as the ratio of pulmonary to systemic flows, failed to show comparable correlations with clinical symptoms. Respiratory rate was related to levels of norepinephrine (r = 0.47***, p < 0.001) and plasma renin activity (r = 0.65***, p < 0.001). The important impact of autonomic imbalance on respiratory rate was underlined by an analysis of variability of heart rate in 26 infants that showed significantly reduced values for the domains of time and frequency. We were not able to find a conclusive multiple regression model with which to explain the symptom "failure to thrive". CONCLUSIONS: A increased heart rate, reduced variability in heart rate, and elevated levels of norepinephrine and renin are significant predictors of clinical symptoms such as tachypnea in infants with congenital cardiac malformations. The neurohormonal hypothesis, in which heart failure is interpreted not only as a hemodynamic derangement but also as a neurohormonal disorder, may be valid for infants with congenital cardiac malformations. PMID- 11727905 TI - Bubble contrast echocardiography in detecting pulmonary arteriovenous malformations after modified Fontan operations. AB - The development of pulmonary arteriovenous malformations is a well-known complication after Fontan operations, and may result in significant morbidity due to increasing arterial desaturation. We compared the use of bubble contrast echocardiography and pulmonary angiography in detecting such malformations. We also examined which anatomical and haemodynamic variables were associated with their development. Our study includes 20 patients who had undergone modified Fontan procedures, 10 with atriopulmonary and 10 with total cavopulmonary connections, in Gothenburg between 1980 and 1991. All patients underwent cardiac catheterisation and pulmonary angiography. Bubble contrast echocardiography was performed at the same time, with injection of agitated polygelin colloid solution (Haemaccel, Hoechst) into the right and left pulmonary arteries, respectively. Transoesophageal echocardiography was used to detect the appearance of bubble contrast in the pulmonary venous atrium. The aim was also to evaluate the role of hepatic venous blood. Of the 20 patients, 9 (45%) had a positive contrast echocardiography study, compared with only 2 (10%) detected by pulmonary angiography. Patients with positive contrast echocardiography had a significantly lower arterial oxygen saturation than those with negative studies, both at rest (88% vs 95%, p < 0.01) and during exercise testing (78% vs 89%, p = 0.01). Bubble contrast echocardiography is much more sensitive in detecting pulmonary arteriovenous malformations than pulmonary angiography. By injecting echo contrast into the right and left pulmonary arteries, the method can be made highly selective. Pulmonary arteriovenous malformations develop much more frequently in patients with the Fontan circulation than previously reported. PMID- 11727906 TI - Quantification of morphologic and hemodynamic severity of coarctation of the aorta by magnetic resonance imaging. AB - OBJECTIVE: As the morpholgic severity of coarctation of the aorta is difficult to assess, especially after previous repair, the value of the technique of multiplanar reconstruction of magnetic resonance imaging data to achieve a 3 dimensional reconstruction of the aortic arch was evaluated and compared to hemodynamic measurements. METHODS AND RESULTS: We performed 30 examinations in 27 patients aged from 6 to 54 years, with a mean of 21 years, by magnetic resonance imaging using a 1.5Tesla scanner with a standard body coil. Measurements of flow across the coarctation were performed using phase shift velocity mapping, and peak velocity was calculated at the site of stenosis. Aortic cross-sectional area before, at, and beyond the stenosis was reconstructed 3-dimensionally to calculate a percentage degree of stenosis. Morphologic severity of stenosis was correlated to invasively assessed hemodynamic gradients and morphologic data from biplane angiography in 23 patients. Among the 30 examinations, 24 patients had been previously treated by either surgery, in 17 patients, or balloon dilation, while 6 had native coarctation. 3-dimensional reconstruction was possible in all and better delineated the anatomy concerning the hemodynamic relevance of stenoses even as compared with biplane angiography. The correlation between severity of narrowing assessed by diameter measurements in the biplane angiography and 2-dimensional magnetic resonance imaging was r = 0.94, and multiplanar reformation with 2-dimensional magnetic resonance imaging was r = 0.87 with a tendency of higher grading with the 3-dimensional technique (p = 0.0001). The correlation of 2-dimensional magnetic resonance imaging with invasively measured hemodynamic gradients was r = 0.67 versus r = 0.74 for the areas assessed by multiplanar reformation, indicating that the hemodynamic relevance of a morphological approach to evaluate the degree of a stenosis should better be assessed 3-dimensionally. CONCLUSIONS: The 3-dimensional reconstruction of the morphologic severity of coarctation offers additional information over conventional imaging especially in patients with kinking, complex geometry, or collaterals, in whom hemodynamic measurements can become unreliable. PMID- 11727907 TI - Influence of the introduction of Amplatzer device on the interventional closure of defects within the oval fossa in children. AB - Since June 1998, we have used an Amplatzer device whenever considered appropriate in patients with isolated defects within the oval fossa. The aim of this study was to define the total cohort of patients with isolated defects in the oval fossa seen at this hospital, so as to assess the impact of this policy on contemporary management. In the two-year period commencing 1st June 1998, 116 patients older than 6 months were seen with an isolated septal defect within the oval fossa. Mean age at closure or last review was 5.8 years, with a range from 0.5 to 20 years. In total, 42 (36%) patients were assigned to surgical closure, 25 (22%) to closure using an Amplatzer device, and 49 (42%) remained under clinical follow up. Direct referral for surgical closure occurred in 24 (21%) patients, in whom transcatheter closure was considered not appropriate after transthoracic echocardiography. Transoesophageal echocardiography was performed in 45 (39%) patients to assess suitability for closure using the Amplatzer device. Of these, 20 (44% of the group undergoing transoesophageal echocardiography) were considered unsuitable for closure in this fashion. Of these, 18 were referred for surgery and 2 with small defects were considered not to require closure. Patients undergoing closure with the device were older than the group referred for surgical closure, having a median age of 7.8 versus 3.6 years, and stayed for a shorter period in hospital. Those closed using the device stayed for 2 days, as opposed to a median of 5 days, with a range from 4 to 10 days for those undergoing surgical closure. Closure was complete as assessed by echocardiography after follow up of 1-3 months in both groups. There were no recognised complications related to insertion of the device, whereas transient postoperative morbidity occurred in 38% of those closed surgically. Insertion of an Amplatzer device was considered to be appropriate in 37% of patients older than 6 months requiring closure of an atrial septal defect in the oval fossa. PMID- 11727908 TI - A national network for the tele-education of Canadian residents in pediatric cardiology. AB - A trial of 11 video-conferenced teaching sessions for residents in pediatric cardiology was performed by the 7 training programs in Canada in order to share expertise in specialized areas, to expose trainees to educational telemedicine, and to acquaint residents with other programs and personnel. Topics included cardiac pathology, arrhythmias, magnetic resonance imaging, fetal physiology, pulmonary hypertension, and cardiomyopathy. The sessions were evaluated by 93 residents by questionnaire for content and technology. Session content was highly rated. Videoconference picture quality was highly rated, but sound quality and visual aids were rated as neutral or unsatisfactory by a significant minority, related to problems with several early sessions, subsequently corrected. 60% of respondents rated the videoconferences as good as live presentations. Presenters were generally satisfied although they required some adjustments to videoconferencing. The average cost per session was $700 Canadian. Videoconferencing of resident educational sessions was generally well accepted by most presenters and residents, and the trial has formed the basis for a national network. Adequate organizational time, and careful attention to audiovisual needs, are most important. Videoconference guidelines are suggested for presenters based on this experience. PMID- 11727909 TI - Young Investigator's Prizewinner 2001. Direct visualization of the influence of normothermic as opposed to hypothermic cardiopulmonary bypass on the systemic microcirculation in neonatal piglets. AB - The direct visualization of systemic microcirculation using intravitalmicroscopy permits the classification of proinflammatory and ischemic microvascular alterations during normothermic and hypothermic cardiopulmonary bypass in neonates. We used seven newborn piglets, on average aged 9 days, and weighing 3200g, as a control group. In addition, we studied nine piglets subjected to 60 minutes of constant nonpulsatile flow using hypothermic extracorporeal circulation at 28 degrees C, and five piglets using normothermic conditions at 37 degrees C. The microvascular network of the greater omentum and the subcutaneous tissue was directly visualized using intravitalmicroscopy. We analysed interactions between leukocytes and endothelial cells, microvascular morphology, and microrheological conditions, focussing on signs of ischemic and proinflammatory alterations. During normothermic cardiopulmonary bypass, the numbers of activated leukocytes were elevated compared to hypothermic cardiopulmonary bypass (p > 0.05). Arteriolar diameter decreased during hypothermia. Capillaries were markedly dilated during normothermia. Patterns of microvascular perfusion, for both types of cardiopulmonary bypass, showed signs of ischemic damage, revealed by a reduced functional capillary density. Perfusion dependent levels of lactate were higher during normothermic cardiopulmonary bypass (p > 0.05). This new experimental approach revealed that non-pulsatile cardiopulmonary bypass, independent of temperature, induces a proinflammatory and ischemic response compared to an unaltered control group. The markedly elevated numbers of activated adherent leukocytes, the reduced capillary density, and the high lactate levels in those undergoing bypass in normothermic conditions indicate a more pronounced inflammatory stimulus and tissue hypoperfusion compared to the possible protective effect of hypothermia for children undergoing cardiopulmonary bypass. PMID- 11727910 TI - Sarcoma of the mitral valve causing coronary arterial occlusion in children. AB - Primary tumors of the cardiac valves are rare. One of the most common reasons that left-sided cardiac tumors come to clinical attention is embolization to the systemic circulation. We present two children who suffered left coronary arterial occlusion due to embolization of a sarcoma of the mitral valve. A 6-year-old female who had been admitted to the hospital after cerebrovascular embolization of a fragment of sarcoma of the mitral valve experienced sudden cardiovascular collapse due to occlusion of the left coronary artery. She was placed on extracorporeal membrane oxygenation, and underwent coronary embolectomy and resection of the tumor from the mitral valve and its tendinous cords. Left ventricular function did not improve, and she underwent orthotopic heart transplantation. On follow-up 32 months after transplant, the patient is well, with no evidence of recurrence of or metastasis from the tumor. The tumor arose from the leaflets and tendinous cords of the mitral valve, and was composed grossly of multiple white nodules. Histopathologic evaluation disclosed fragments composed predominantly of peripheral spindle cells in an extensive fibromyxoid stroma. The mildly pleomorphic cells of the tumor gradually blended with adjacent pieces of the mitral valvar leaflet and tendinous cords. Immunohistochemical studies revealed strong staining for vimentin, smooth muscle actin, muscle specific actin, and myoglobin, suggesting myogenic differentiation. The other patient was a 2 1/2-year-old female who died suddenly at home. Grossly and histologically, the tumor was essentially identical to the first case, and there was a 3 cm string-like extension passing into the orifice of the left coronary artery. To put the cases in context, we compare them with other descriptions of this rare type of tumor. PMID- 11727911 TI - Discordances between the different types of atrial arrangement and the positions of the thoraco-abdominal organs. AB - Numerous investigations have addressed the most appropriate anatomical structures to be used for the identification of segmental arrangement ("situs"). Variations from normal have been termed "heterotaxy syndromes". The assumption is frequently made that, in such instances, all the organs in a given individual case follow the same rules for segmental morphology. This leads to confusion. The starting point for cardiac analysis is the structure of the atriums. On this basis, we can define viscero-atrial discordance when, in usual arrangement, mirror imagery, and heterotaxy, the atriums and their appendages are no longer concordant with the remaining thoracic and abdominal organs. With this in mind, we examined a large number of autopsied specimens to identify all possible variations of thoracic structures and abdominal organs relative to atrial arrangement. We analyzed 1100 malformed hearts. Of these, 1046 had usual arrangement with normally situated heart (95%); 10 cases had usual arrangement with right-sided heart (0.90%); 25 showed mirror imagery (2.27%), and 19 were found with heterotaxy (1.72%). There was a total of 54 hearts of usual arrangement with right-sided heart, mirror imagery and heterotaxy, (4.90%) and, amongst them, 17 showed viscero-atrial discordances (31.48%). In those having usual arrangement with right-sided heart, there were 2 cases (20%); 8 in those with mirror imagery (32%); and 7 in heterotaxy (36.8%). In the 1046 cases of usual arrangement with normally situated heart, there were 12 examples of viscero-atrial discordances (1.14%). In all, we found a total of 29 cases of viscero-atrial discordances. Taken together, the discordances are best dealt with by describing separately, in every case of normal or abnormal atrial arrangement, the thoracic structures, the atrial appendages, and the abdominal organs. In the final analysis, the diagnosis of the cardiac arrangement rests on the location and morphology of the atriums and their appendages. PMID- 11727912 TI - Lucio Parenzan. PMID- 11727913 TI - Tricuspid valvar endocarditis caused by penicillin resistant pneumococcus. AB - A 6-year-old boy presented with endocarditis of the tricuspid valve caused by penicillin resistant pneumococcus. He had no history of congenital cardiac disease. The likely portal of entry for the organism was the middle ear. PMID- 11727914 TI - Pseudoachondroplasia in a child with prolapse of the mitral valve. AB - Pseudoachondroplasia is an autosomal dominant variant of osteochondroplasia that results in mild to severe short-limb dwarfism and early-onset of osteoarthrosis. It has been linked to results from mutations in the gene for cartilage oligomeric matrix protein. We describe a 4-year-old boy with pseudoachondroplasia who also had prolapse of the mitral valve. To the best of our knowledge, this association has not previously been reported. PMID- 11727915 TI - Use of tissue harmonic imaging in pediatric echocardiography. AB - Tissue harmonic imaging improves the quality of images in adults with poor acoustic windows, but its utility in children has yet to be evaluated. The use of fundamental and harmonic imaging were compared in 70 children with poor echocardiographic windows. Scores were higher for harmonic in comparison to fundamental imaging for all views. Harmonic imaging improves the visualization of cardiac structures over fundamental imaging in children with poor echocardiographic windows. PMID- 11727917 TI - Surgical repair of naturally palliated anomalous origin of the left coronary artery from the right pulmonary artery. AB - Anomalous origin of the left coronary artery from the pulmonary arteries is a rare and life-threatening defect, usually needing prompt surgical correction during infancy. We describe the case of a young asymptomatic patient with this defect who underwent surgical reimplantation despite the absence of signs of myocardial ischemia, due to the presence of proximal stenosis of the anomalous coronary artery. PMID- 11727916 TI - Treatment of rheumatic carditis with intravenous gammaglobulin: is there a beneficial effect? AB - Rheumatic carditis is a major manifestation of acute rheumatic fever. Conventional therapy includes the use of salicylates and steroids. To date, however, such therapy has not been proven to have a clear benefit in reducing valvar heart disease. We report the use of high-dose intravenous immunoglobulin in two chidlren with acute rheumatic carditis in whom we have been able to document the beneficial effect. PMID- 11727918 TI - Transcatheter occlusion of a large coronary arterial fistula with new detachable platinum microcoils. AB - We report a 17 year old boy with a large fistula from the right coronary artery to the right ventricle. The fistula consisted of two major branches. The smaller branch could be embolised with three detachable platinum microcoils without problems. The high flow of blood in the larger branch, however, prevented conventional occlusion using coils. Interventional occlusion of this branch was achieved in a second attempt. Following creation of an arterio-venous wire-loop, we advanced a balloon catheter into the distal end of the fistula. Under temporary occlusion of the fistula, complete closure was achieved by delivery of 7 detachable platinum microcoils. Thus, even large coronary arterial fistulas can be closed safely with these new platinum detachable microcoils. The procedure, however, requires temporary control of the flow of blood by balloon occlusion. PMID- 11727919 TI - Use of a cutting balloon catheter to dilate resistant stenoses in major aortic-to pulmonary collateral arteries. AB - We report the successful use of a cutting-balloon catheter in dilating stenoses in major systemic-to-pulmonary collateral arteries which had been resistant to high pressure balloons. After dilation using the cutting balloon, we implanted stents to produce a marked improvement in both pulmonary blood flow and arterial saturation. The technique provides useful palliation for patients having such stenotic collateral arteries. PMID- 11727920 TI - Imai technique for management of the coronary arteries arising from a solitary sinus in discordant ventriculo-arterial connections. PMID- 11727921 TI - Hypertrophic cardiomyopathy and myocardial bridging. PMID- 11727922 TI - International registry for ventricular repair of congenital cardiac malformations. PMID- 11727923 TI - The tumour microenvironment: causes and consequences of hypoxia and acidity. Introduction. PMID- 11727924 TI - pH regulation of renal gene expression. AB - The increase in intracellular pH (pHi) associated with various tumour cells triggers changes in gene expression. Similar adaptations also occur as part of the physiological response to changes in acid base balance. For example, during metabolic acidosis, increased renal ammoniagenesis and bicarbonate synthesis are sustained by the increased expression of various transport proteins and key enzymes of glutamine metabolism. In rat kidney, increased expression of the mitochondrial glutaminase (GA) and glutamate dehydrogenase (GDH) results from stabilization of their respective mRNAs. The 3'-untranslated region (UTR) of the GA mRNA contains a direct repeat of an 8-base AU sequence that functions as a pH response element. This sequence exhibits a high affinity and specificity for z crystallin. The same protein binds to two separate, but homologous, 8-base AU sequences within the 3'-UTR of the GDH mRNA. The apparent binding activity of z crystallin is increased significantly during onset of metabolic acidosis. Thus, increased binding of z-crystallin may initiate the pH-responsive stabilization of the two mRNAs. In contrast, induction of the phosphoenolpyruvate carboxykinase (PEPCK) gene occurs at the transcriptional level. In LLC-PK1-FBPase+ kidney cells, a decrease in pHi leads to activation of the p38 stress-activated protein kinase and subsequent phosphorylation of ATF-2. This transcription factor binds to the CRE-1 element within the promoter of the PEPCK gene to enhance transcription. Similar mechanisms may contribute to altered gene expression in tumour cells. PMID- 11727925 TI - Genetic determinants that influence hypoxia-induced apoptosis. AB - The p53 tumour suppressor gene is capable of activating both death receptor and mitochondrial-signalled forms of apoptotic cell death in response to diverse stimuli. Studies have suggested that impairment of the mitochondrial-signalled Apaf/caspase 9 pathway and not the death receptor Fas pathway results in almost complete resistance to apoptotic cell death induced by a low oxygen environment. However, it is unclear how p53 signals the activation of this pathway and whether it is through already identified p53 effector genes such as the pro-apoptotic gene bax, or through novel effectors such as BNIP-3/BNIP-3L. Comparison of cell lines genetically matched at the bax, cytochrome c, apaf, caspase 9 and caspase 3 loci indicated that except for bax, all of these genes were essential for hypoxia induced apoptosis both in cell culture and in transplanted tumours. These data imply that cytochrome c plays a pivotal role in signalling cell death by apoptosis under hypoxic conditions, and that the release of cytochrome c is independent of both Bax and p53. In contrast to cytochrome c, p53 modulates the magnitude of apoptosis under hypoxic conditions, but in itself is not required for the activation of the caspase cascade. PMID- 11727926 TI - Genomic instability in cancer. AB - Solid tumours have abnormal, deficient vascular and lymphatic systems. As a result, perfusion within these malignancies is inadequate and chaotic, and the cancers contain regions that are transiently and chronically exposed to low pH, severe hypoxia and nutrient deprivation. These microenvironmental inadequacies are present from the earliest point in the development of solid tumours, and are fully established while the neoplasms are still microscopic. Exposures to acidic and hypoxic environments have been shown to produce a wide range of cytogenetic changes. These include increases in mutation frequencies; deficits in DNA repair; DNA overreplication and gene amplification; the induction of chromosomal fragile sites, triggering genomic rearrangements; and changes in gene expression. Moreover, exposure of cells to adverse microenvironments such as those in solid tumours selects for cells which have defects in the structure or expression of the genes that normally regulate cell proliferation. The genetic changes and selection pressures induced by hypoxia may be critical in causing the development of the genomic instability and genetic heterogeneity which is characteristic of solid tumours and in fostering the evolution of relatively benign cell populations in solid tumours to increasingly malignant, increasingly aggressive phenotypes. PMID- 11727927 TI - pH, hypoxia and metastasis. AB - The vascular supply to tumours is often poorly formed and irregular with the result that tumours may contain regions of poor nutritional supply with hypoxia and acidic pH. Clinical studies have demonstrated substantial heterogeneity in oxygenation in human tumours. In some studies tumours with poorer oxygenation were more likely to have metastasized. In our studies of carcinoma of the cervix, nodal metastases were 1.5 times more likely at diagnosis in patients with more hypoxic tumours relative to those with less hypoxic tumours. Transplanted rodent KHT fibrosarcomas and SCC-VII squamous cell tumours also have variability in levels of oxygenation; again, the more hypoxic tumours are found to be more metastatic. Furthermore, deliberate exposure of KHT tumours to cyclic hypoxia (12 cycles of 5% oxygen breathing for 10 min interspersed with 10 min air breathing) every day during their growth, doubled the level of micrometastases that were detected in the lungs of the mice. These findings are consistent with in vitro studies demonstrating that KHT and SCC-VII cells and B-16 melanoma cells exposed to hypoxia or low pH have increased propensity to form metastases following injection into-mice. This effect is transient and is lost within about 48 h of removal of exposure to hypoxia or low pH, suggesting that the effect may be due to changes in gene expression associated with that stress. Recent studies have implicated a number of genes, such as vascular endothelial growth factor and interleukin 8, in the effect of hypoxic and acid pH on metastasis. PMID- 11727928 TI - Modelling tumour acidity and invasion. AB - The intracellular pH (pHi) of mammalian cells is tightly regulated by the concerted action of a number of different pumps in the plasma membrane. Despite the acidic extracellular environment (pHe 6.8-7.0) of some tumours, the pHi of solid tumours is neutral or slightly alkaline compared to normal tissues (pHi 7.0 7.4). This gives rise to a reversed pH gradient across the cell membrane between tumours and normal tissue, which has been implicated in many aspects of tumour progression. One such area is tumour invasion: the incubation of tumour cells at low pH has been shown to induce more aggressive invasive behaviour in vitro. In this paper we use mathematical models to investigate whether altered proteolytic activity at low pH is responsible for the stimulation of a more metastatic phenotype. We examined the effect of culture pH on the secretion and activity of two different classes of proteinases: the metalloproteinases (MMPs), and the cysteine proteinases (such as cathepsin B). The modelling predicts that, in addition to metalloproteinase activity, the pH-induced peripheral redistribution of cathepsin B could be a major factor involved in the acquisition of a more metastatic phenotype in malignant cells at low pHe. PMID- 11727929 TI - pHi, aerobic glycolysis and vascular endothelial growth factor in tumour growth. AB - Signalling via growth factors, oncogenes and environmental stresses such as hypoxia, promotes the up-regulation of glycolysis, intracellular pH (pHi) and vascular endothelial growth factor (VEGF) via cooperative mechanisms. Somatic cell genetics was applied to a fibroblastic cell line (CCOL39) to disrupt either aerobic glycolysis, respiration, or a major pHi-regulating system, the Na-H exchanger (NHE1). We obtained stable mutants impaired either in phosphoglucose isomerase (pgi-), which produce virtually no lactic acid, or in respiration (res ), which over secrete lactic acid (three- to fourfold the wild-type rate). These mutations, which allowed us to analyse the incidence of lactic acid production on tumour development in nude mice, were analysed alone, or in combination, with the mutation nhe1- to evaluate in vivo the role of NHE1 on pHi control and cell proliferation. Ras-transformed pgi- cells (not forming lactic acid) form tumours like wild type transformed cells (100% incidence). The disruption of NHE1 however, strongly reduced tumour incidence to about 20%. In cells bearing both mutations, nhe1-, res-, and which therefore over-produce lactic acid, the situation is even more dramatic (0% incidence). In sharp contrast, association of nhe1- with pgi- restored 100% tumour incidence. We conclude that over-production of lactic acid is detrimental for tumour development and that NHE1, by controlling pHi, plays a key role in cell survival/proliferation and tumour growth. Finally we summarize our current knowledge on the signalling mechanisms leading to VEGF expression, another key component of tumour growth via neo vascularization. PMID- 11727930 TI - pH and chemotherapy. AB - In vivo pH measurements by magnetic resonance spectroscopy reveal the presence of large regions of acidic extracellular pH in tumours, with the intracellular pH being maintained in the neutral-to-alkaline range. This acid-outside plasmalemmal pH gradient acts to exclude weak base drugs such as the anthracyclines and vinca alkaloids, a behaviour that is predicted by the decrease in octanol-water partition coefficients of mitoxantrone and doxorubicin with decreasing solution pH. This pH gradient can be reduced or eliminated in mouse models of breast cancer by systemic treatment with sodium bicarbonate. We have demonstrated tumour alkalinization following chronic ad libitum administration of NaHCO3 and acute intraperitoneal administration of NaHCO3 to tumour-bearing mice. Chronic treatment of tumour-bearing SCID mice with NaHCO3 results in an enhancement in MCF-7 tumour xenograft response to doxorubicin. Intraperitoneal administration of NaHCO3 to tumour-bearing C3H/Hen mice prior to treatment with mitoxantrone results in a greater-than 4.5-fold increase in cell-kill in the syngeneic C3H mammary tumour model. Most combination chemotherapy regimens include at least one weak base drug. Our results suggest that agents such as sodium bicarbonate, Carbicarb and the diuretic furosemide--which are known to induce metabolic alkalosis in humans--may be useful in enhancing the efficacy of these treatment regimens in humans. PMID- 11727931 TI - The HIF pathway: implications for patterns of gene expression in cancer. AB - Regulation of the growth and metabolism of large organisms is tightly constrained by the need for precise oxygen homeostasis. Work on control of the haematopoietic growth factor erythropoietin has led to the recognition of a widespread transcriptional response to hypoxia which provides insights into how this is achieved. The central mediator of this response is a DNA binding complex termed hypoxia inducible factor 1 (HIF-1), which plays a key role in the regulation by oxygen of a large and rapidly growing panel of genes. In cancer, activity of the HIF system is up-regulated both by microenvironmental hypoxia and by genetic changes. The clearest example of genetic activation is seen in the hereditary cancer syndrome von Hippel-Lindau (VHL) disease. In normal cells the product of the VHL tumour suppressor gene targets the regulatory HIF subunits (HIF-1alpha and HIF-2alpha) for oxygen-dependent proteolysis, acting as the substrate recognition component of an E3 ubiquitin ligase. In pVHL defective cells this process is blocked leading to constitutive up-regulation of HIF-1alpha subunits, activation of the HIF complex and overexpression of HIF target genes. Using gene array screens we have defined a large number of VHL-regulated genes. The majority of these show hypoxia-inducible responses, supporting the central involvement of pVHL in gene regulation by oxygen. In addition to known HIF target genes involved in angiogenesis, glucose metabolism and vasomotor control, these new targets include examples with functions in matrix metabolism, apoptosis, carbon dioxide metabolism and secondary cascades of transcriptional control. Thus activation of HIF provides insights into the classical metabolic alterations in cancer cells, and into the mechanisms by which microenvironmental hypoxia might influence tumour behaviour. In the case of VHL disease, this activation can be linked to mutations in a defined tumour suppressor gene. Equally regulation of the HIF 1alpha/pVHL interaction in normal cells should provide insights into the physiological mechanisms operating in cellular oxygen sensing. PMID- 11727932 TI - The physiological environment in cancer vascularization, invasion and metastasis. AB - One of the most lethal aspects of cancer arises from its ability to invade and metastasize. Determining the factors that promote cancer cell invasion and metastasis is therefore critically important in treating this disease. The tumour physiological environment is uniquely different from normal tissue, and exhibits hypoxia, acidic extracellular pH and high levels of lactate. This environment, dictated largely by abnormal tumour vasculature and metabolism, in turn also promotes angiogenesis. The physiological environment, tumour metabolism, angiogenesis and vascularization are therefore inextricably linked. We have developed and applied non-invasive magnetic resonance (MR) imaging (I) and spectroscopy (S) techniques to understand the role of vascular, physiological and metabolic properties in cancer invasion and metastasis. These MR studies are performed with human breast and prostate cancer cells maintained in culture or grown as solid tumours in immune-suppressed mice. We have detected significant differences in vascular, physiological and metabolic characteristics of metastatic and non-metastatic human breast and prostate cancer models with MRI and MRS. Using a combined MRI/MRS approach we are currently acquiring metabolic, extracellular pH and vascular images from the same localized regions within a solid tumour to further understand the dynamics between these parameters and their role in cancer invasion and metastasis. PMID- 11727933 TI - pH and multidrug resistance. AB - Most chemotherapeutic drugs in use today are hydrophobic small molecules that are also typically either weakly basic, weakly acidic or charged. Thus, changes in the electrochemical parameters of tumour cell membranes have important effects on their transmembranous diffusion and cellular retention. Changes in these parameters can also modulate the function of immunological agents, and affect the signal transduction associated with induction of apoptosis. For these reasons, it is logical to propose that many (if not most) of the characteristics of multidrug resistant (MDR) tumour cells could be due to perturbations in cellular ion transport. Indeed, many reports of altered ion transport in MDR cells can be found in the literature. Moreover, many studies suggest that P glycoprotein (Pgp) overexpression confers this altered ion transport, however, detailed physical chemical analysis of this phenomenon has been confused by the complexity of the model systems devised to study Pgp. To help resolve this confusion, our laboratory has focused on a detailed characterization of 'pure' and stable Pgp transfectants unadulterated by the complications of chemotherapeutic drug exposure, various yeast strains and yeast vesicle preparations, and purified, reconstituted Pgp preparations. Recent data obtained with these model systems are summarized in this paper. PMID- 11727934 TI - 'The metabolism of tumours': 70 years later. AB - Otto Warburg's classic treatise on the reprogramming of tumour metabolism from oxidative to glycolytic metabolism was published in London in 1930. Although the Warburg effect is one of the most universal characteristics of solid tumours, the molecular basis for this phenomenon has only recently been elucidated by studies indicating that increased expression of genes encoding glucose transporters and glycolytic enzymes in tumour cells is mediated by the transcription factors c-MYC and HIF-1. Whereas c-myc is a direct target for oncogenic mutations, expression of hypoxia-inducible factor 1 (HIF-1) is indirectly up-regulated via gain-of function mutations in oncogenes and loss-of-function mutations in tumour suppressor genes that result increased HIF-1alpha protein expression and/or increased HIF-1 transcriptional activity in a cell-type-specific manner. As a result of genetic alterations and intratumoral hypoxia, HIF-1alpha is overexpressed in the majority of common human cancers relative to the surrounding normal tissue. In human breast cancer and brain tumours, HIF-1alpha overexpression is strongly correlated with tumour grade and vascularity. PMID- 11727935 TI - The multiple mechanisms of multidrug resistance and cellular pH. AB - Multidrug resistance (MDR) describes a variety of strategies that tumour cells develop to evade the cytotoxic effects of anti-cancer drugs. Tumour cells that have become MDR show decreased cellular sensitivity to the drug employed in chemotherapy as well as a broad spectrum of drugs without obvious common targets or structural homology. MDR is a major obstacle to the successful treatment of tumours. Our understanding of multidrug resistance is limited, in part, by our limited knowledge of what makes tumour cells more sensitive to chemotherapeutics. Drug resistance may subvert the same mechanisms that make tumour cells hypersensitive or use molecular modifications unrelated to the mechanisms that lead to hypersensitivity in tumour cells. Indeed, multidrug resistance is likely to be the consequence of a multitude of mechanisms. Some drug-resistant and drug sensitive mammalian cell lines have shown aberrations in regulation of cellular pH. These changes can contribute to some of the physiological and cell biological changes that are observed both in drug-sensitive and MDR tumour cell lines. As such, at least in some tumour cell lines, the changes of pH contribute both to the physiology of transformation and the physiology of drug-resistance. The observation that these variations from normal pH are not universally observed in either tumour or MDR cells suggests that these mechanisms cannot be generalized to all transformed or MDR cells. It remains to be resolved to what extent these changes are a consequence of studying cell lines grown in vitro or if they reflect the in situ physiology of human tumours. There are no detectable changes in cytosolic or organellar pH in cells transfected with P-glycoprotein, a plasma membrane-based protein implicated in drug resistance. Thus, there are likely to be multiple mechanisms contributing to multiple drug-resistance. PMID- 11727936 TI - Why are cancers acidic? A carrier-mediated diffusion model for H+ transport in the interstitial fluid. AB - 31P MRS studies have shown that the intracellular compartment ot tumours is kept near neutrality, whereas the interstitial fluid is acidic (pH 6.5-6.8). Why is this compartment acidic? Balance studies confirm that tumours produce excessive lactic acid, although less than usually supposed, but this cannot be the whole story, since Tannock and co-workers have shown interstitial acidity in glycolysis deficient tumours. Another major acid load is caused by hydration of CO2 molecules to carbonic acid, catalysed by carbonic anhydrase. The distance that H+ must diffuse from cancer cells to capillaries is further than in normal tissue and this will increase acidification near the cells. We show that previous quantitative models based on simple H+ diffusion are unsatisfactory. This is because most H+ ions cross the interstitial space bound to buffers such as inorganic phosphate. Although these protonated buffers (i.e. conjugate acids) diffuse much more slowly than H+ ions they carry most of the protons, so the pH predicted by this model is closer to neutrality for a given proton production rate than that predicted by the dissolved H+ model. We have developed a mathematical model of this carrier-mediated system that predicts pHe values as low as those observed in some tumours. PMID- 11727937 TI - pH and therapy of human cancers. AB - Studies in model systems have demonstrated that tumour pH can be a determinant of treatment response. The potential that tumour pH differs from that of normal tissues may provide a basis for selective killing of tumour cells. Although the data are limited, pH measurements in humans indicate a difference between tumour and normal tissues. In general, electrode pH (generally considered to reflect primarily extracellular pH, pHe) is lower in tumour than normal tissue. However, pH measured by magnetic resonance spectroscopy (MRS) or positron emission tomography (PET; both are generally considered to reflect primarily intracellular pH, pHi) is equal to or slightly higher in tumours than normal tissues. Hence, not only may pHe and pHi differ between normal and malignant tissues, but the pH gradient (which determines the distribution of chemotherapeutic agents that are weak acids or bases) is also reduced or reversed in tumours. To date, the majority of treatment-related studies conducted have focused on hyperthermia (combined with radiotherapy) due to the recognized importance of acidic pH as a thermal sensitizer. However, the results have been somewhat surprising: patients with a better response to hyperthermia radiotherapy have higher pH (as measured by electrode or 31P MRS) prior to treatment. PMID- 11727938 TI - Frontiers in the measurement of cell and tissue pH. AB - Investigation into the causes and consequences of aberrant pH regulation is entirely dependent on the ability to measure this parameter with specificity and sensitivity. Since their beginnings in the 1950s, techniques for measuring cell and tissue pH have undergone a number of significant advances. Following each of these advances, new investigators have been brought into the field, and new discoveries have been made. From microelectrode and dye distribution studies, measurement of pH underwent a revolution with the advent of pH-sensitive dyes that could be loaded into the cytosol. A further significant advance came from the measurement of cell and tissue pH in whole organisms by magnetic resonance spectroscopy (MRS). Frontiers in pH measurement exist in the development of pH sensitive proteins and pH-sensitive MR contrast agents. These are predicted to bring even more people into this fascinating field, and generate more important discoveries. PMID- 11727939 TI - Mathematical models of tumour invasion mediated by transformation-induced alteration of microenvironmental pH. AB - The invasive phenotype, characteristic of malignancy, is found in multiple transformed populations with widely varying genotypes. The authors hypothesize that this genetic heterogeneity and instability precludes mechanisms of tumour invasion requiring consistent, coordinated production of one or more proteins or other macromolecules. Instead, this model assumes that the genetically disordered, unstable populations found in tumours must employ a simple mechanism of invasion that arises from one common trait of all transformed cells--reversion to a phenotype more primitive (less differentiated) than the tissue of origin. Specifically, this approach focuses on primitive metabolic pathways with preferential use of glycolysis for energy production. Mathematical models of invasive cancer based on tumour-induced acidification of the microenvironment are presented. Using population ecology and diffusion-reaction models, it is shown that normal tissue adjacent to the tumour edge is subjected to an extracellular pH that is significantly lower than normal. This leads to degradation of the interstitial matrix, loss of intercellular gap junctions and cell necrosis. Tumour cells have an optimal extracellular pH less than that of normal cells and are thus able to thrive in the acidic microenvironment expanding into the space of the dying normal tissue. The model is consistent with extant data on the tumour microenvironment as well as clinical data relating increasing tumour invasiveness with elevated glucose utilization and lactate content. It predicts well-established phenomena in tumorigenesis such as the adenoma-carcinoma sequence and the critical role of angiogenesis in the invasive phenotype. An acellular gap at the tumour host interface is also predicted and can be demonstrated in pathologic specimens. Novel treatment approaches based on this model are discussed. PMID- 11727940 TI - Parental drinking problems and children's adjustment: vagal regulation and emotional reactivity as pathways and moderators of risk. AB - Children's emotional regulation (as indexed by vagal suppression) and children's emotional reactivity during an argument were examined as moderators and mediators of parental problem drinking and children's adjustment in a sample of 6- to 12 year-olds. Cardiac vagal tone was assessed during both a baseline condition and exposure to an audiotaped argument. Vagal suppression was calculated by subtracting vagal tone during the baseline from that recorded during the argument, with a higher number representing increased suppression of vagal tone during the argument. Emotional reactivity was based on both observations of overt behaviors of children and their reported feelings during the argument. A higher level of vagal suppression was a protective factor against children's externalizing, internalizing, and social problems associated with exposure to parental problem drinking. Emotional reactivity was a vulnerability factor, and children's increased anger and fear, and to a lesser degree sadness, each moderated and exacerbated the effects of parental problem drinking on child outcomes. PMID- 11727941 TI - Sources of covariation among attention-deficit/hyperactivity disorder, oppositional defiant disorder, and conduct disorder: the importance of shared environment. AB - Research has documented high levels of covariation among childhood externalizing disorders, but the etiology of this covariation is unclear. To unravel the sources of covariation among attention-deficit/hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), and conduct disorder (CD), the authors studied 11-year-old twins (N = 1,506) from the Minnesota Twin Family Study. Symptom counts for each of these disorders were obtained from interviews administered to the twins and their mothers. A model was fit that allowed the parsing of genetic, shared environmental (factors that make family members similar to each other), and nonshared environmental (factors that make family members different from each other) contributions to covariation. The results revealed that although each disorder was influenced by genetic and environmental factors, a single shared environmental factor made the largest contribution to the covariation among ADHD, ODD, and CD. PMID- 11727942 TI - Dissociations of the night: individual differences in sleep-related experiences and their relation to dissociation and schizotypy. AB - The author examined the associations among sleep-related experiences (e.g., hypnagogic hallucinations, nightmares, waking dreams, and lucid dreams), dissociation, schizotypy, and the Big Five personality traits in 2 large student samples. Confirmatory factor analyses indicated that (a) dissociation and schizotypy are strongly correlated-yet distinguishable-constructs, and (b) the differentiation between them can be enhanced by eliminating detachment/depersonalization items from the dissociation scales. A general measure of sleep experiences was substantially correlated with both schizotypy and dissociation (especially the latter) and more weakly related to the Big Five. In contrast, an index of lucid dreaming was weakly related to all of these other scales. These results suggest that measures of dissociation, schizotypy, and sleep-related experiences all define a common domain characterized by unusual cognitions and perceptions. PMID- 11727943 TI - Specificity of Stroop interference in patients with pain and PTSD. AB - The authors investigated processing of threat words in motor vehicle accident survivors using a modified Stroop procedure. Three samples were included: 28 participants with comorbid posttraumatic stress disorder (PTSD) and pain, 26 participants with pain without PTSD, and 21 participants without pain or any psychiatric conditions. Four word categories were used: (a) accident words, (b) pain words, (c) positive words, and (d) neutral words. This study examined whether processing biases would occur to accident words only in participants with PTSD or if these biases would also be noted in the No PTSD/Pain sample. Additionally, this study examined whether processing biases would be noted to pain words in the 2 pain samples, irrespective of PTSD. Overall, color naming was significantly slower in the PTSD/Pain group in comparison with the other groups. As well, the PTSD/Pain sample showed significant response delays to both accident and pain-related words, whereas patients with No PTSD/Pain showed delays to pain stimuli only. PMID- 11727944 TI - Negative affect and interpersonal rejection among prison inmates in a therapeutic community: a social relations analysis. AB - The authors used D. A. Kenny's social relations model to examine J. C. Coyne's interpersonal theory of depression among a clinical sample of well-acquainted prison inmates. Members of 12 therapy groups (N = 142) diagnosed with a substance abuse disorder completed a self-report measure of depression and anxiety and indicated their desire to interact with other group members. There was both consensus about which group members were rejected and individual differences in the participants reported desire for future interaction with other group members. Those reporting high levels of depressive negative affect were most likely to be rejected. Those lowest in positive affect indicated the least desire for future interaction with others. PMID- 11727945 TI - Postdisaster stress in the United States and Mexico: a cross-cultural test of the multicriterion conceptual model of posttraumatic stress disorder. AB - Data on symptoms of posttraumatic stress disorder (PTSD) were collected 6 months after Hurricanes Paulina (N = 200; Mexico) and Andrew (non-Hispanic n = 270; United States) using the Revised Civilian Mississippi Scale. A 4-factor measurement model that represented the accepted multicriterion conceptualization of PTSD fit the data of the U.S. and Mexican samples equally well. The 4 factors of Intrusion, Avoidance, Numbing, and Arousal correlated significantly and equivalently with severity of trauma in each sample. A single construct explained much of the covariance of the symptom factors in each sample. However, modeling PTSD as a unidimensional construct masked differences between samples in symptom severity. With severity of trauma controlled, the Mexican sample was higher in Intrusion and Avoidance, whereas the U.S. sample was higher in Arousal. The results suggest that PTSD is a meaningful construct to study in Latin American societies. PMID- 11727946 TI - Clinical risk factors for the generation of life events in major depression. AB - This study examines the relationship of anxiety disorder and dysthymia comorbidity to the generation of life events prior to major depression episode onset in a cross-sectional community sample of 76 women. Those with comorbid anxiety and dysthymia experienced higher rates of events that were at least partly dependent on their own behavior but did not differ from those without these clinical risk factors on independent life events outside of their control. This relationship remained significant even after controlling for overall severity of depression and demographic covariates. The implications of these results for understanding the increased rates of major depression onset and recurrence among those with comorbid anxiety and dysthymia are discussed as avenues of future research. PMID- 11727947 TI - Long-term caregiving: what happens when it ends? AB - Data from a longitudinal study were used to examine what happens to caregivers in the years after their cognitively impaired spouse dies. Comparisons of 42 current caregivers, 49 former caregivers, and 52 noncaregivers over a 4-year period showed that former caregivers did not improve on several measures of psychological well-being. Although former caregivers experienced decreases in stress and negative affect, their scores on depression, loneliness, and positive affect did not rebound to levels comparable to noncaregivers and, in fact, remained similar to those of current caregivers up to 3 years after caregiving had ceased. The most consistent predictors of postcaregiving outcomes were social support and intrusive-avoidant thinking about caregiving. The data suggest that some consequences of long-term caregiving may be long-term as well. The needs of former spousal caregivers warrant greater attention both in research and in practice. PMID- 11727948 TI - Current and lifetime comorbidity of the DSM-IV anxiety and mood disorders in a large clinical sample. AB - The comorbidity of current and lifetime DSM-IV anxiety and mood disorders was examined in 1,127 outpatients who were assessed with the Anxiety Disorders Interview Schedule for DSM-IV: Lifetime version (ADIS-IV-L). The current and lifetime prevalence of additional Axis I disorders in principal anxiety and mood disorders was found to be 57% and 81%, respectively. The principal diagnostic categories associated with the highest comorbidity rates were mood disorders, posttraumatic stress disorder (PTSD), and generalized anxiety disorder (GAD). A high rate of lifetime comorbidity was found between the anxiety and mood disorders; the lifetime association with mood disorders was particularly strong for PTSD, GAD, obsessive-compulsive disorder, and social phobia. The findings are discussed in regard to their implications for the classification of emotional disorders. PMID- 11727949 TI - Adolescence-limited versus persistent delinquency: extending Moffitt's hypothesis into adulthood. AB - The authors examined how neuropsychological, personality, and environmental risk factors and their interactions were related to trajectories of delinquent behavior from adolescence to adulthood. Four waves of longitudinal data from 698 male participants, ages 12-18 at Time 1 and ages 25-31 at Time 4, were included in the analyses. Using a growth mixture model approach, 4 trajectories were identified: nondelinquents, adolescence-limited delinquents, adolescence-to adulthood-persistent delinquents, and escalating delinquents. Five risk factors distinguished escalating from persistent delinquents and 5 also distinguished nondelinquents from the 3 delinquency trajectories. Persistent delinquents scored significantly higher than adolescence-limited delinquents on only one risk factor, disinhibition. Overall, few of the factors that are related to childhood to-adolescence persistence were associated with persistence in delinquency beyond adolescence. PMID- 11727950 TI - Disinhibitory psychopathology in male adolescents: discriminating conduct disorder from attention-deficit/hyperactivity disorder through concurrent assessment of multiple autonomic states. AB - T. P. Beauchaine recently proposed a model of autonomic nervous system functioning that predicts divergent patterns of psychophysiological responding across disorders of disinhibition. This model was tested by comparing groups of male adolescents with attention-deficit/hyperactivity disorder (ADHD) and attention-deficit/hyperactivity disorder plus conduct disorder (CD/ADHD) with controls while performing a repetitive motor task in which rewards were administered and removed across trials. Participants then watched a videotaped peer conflict. Electrodermal responding (EDR), cardiac pre-ejection period (PEP), and respiratory sinus arrhythmia (RSA) were monitored. Compared with controls, the ADHD and CD/ADHD participants exhibited reduced EDR. The CD/ADHD group was differentiated from the ADHD and control groups on PEP and from the control group on RSA. Findings are discussed in terms of the motivational and regulational systems indexed. Implications for understanding rates of comorbidity between CD and ADHD are considered. PMID- 11727951 TI - Exploring gene-environment interactions: socioregional moderation of alcohol use. AB - Examples of gene-environment interaction in human behavioral data are relatively rare; those that exist have used simple, dichotomous measures of the environment. The authors describe a model that allows for the specification of more continuous, more realistic variations in environments as moderators of genetic and environmental influences on behavior. Using data from a population-based Finnish twin study, the authors document strong moderating effects of socioregional environments on genetic and environmental influences on adolescent alcohol use, with nearly a five-fold difference in the magnitude of genetic effects between environmental extremes. The incorporation of specific environmental measures into genetically informative designs should prove to be a powerful method for better understanding the nature of gene-environment interaction and its contribution to the etiology of behavioral variation. PMID- 11727952 TI - Predicting recurrence of major depressive disorder in young adults: a prospective study. AB - Sixty-five young adults with remitted major depressive disorder (MDD) were followed for 18 months. Recurrence of MDD was reported by 41.5% of the initial sample and 49.1% of those who completed the study (n = 53). Survival analyses were used to identify predictors of recurrence so that individuals at greatest risk could be targeted for intervention. Potential predictors included measures of comorbid psychopathology (Axis II pathology, and current and lifetime nonmood Axis I diagnoses), depression-specific clinical features (number of episodes, past treatment, and suicidality), and self-reported cognitive and interpersonal constructs (hope, dysfunctional attitudes, and interpersonal problems). Only personality pathology (specifically, the total dimensional and Cluster B dimensional scores on the International Personality Disorder Examination) significantly predicted hazard of recurrence. PMID- 11727953 TI - Response perseveration in psychopathic women. AB - Recent attempts to validate the Psychopathy Checklist-Revised (PCL-R) as a measure of psychopathy in female offenders have been limited by a failure to examine laboratory correlates of the syndrome. We assessed 112 incarcerated women by using the PCL-R and examined their performance on a card perseveration task that has been used to demonstrate response perseveration in psychopathic men. Contrary to prediction, psychopathic women did not perseverate responding when the PCL-R was used either dimensionally or categorically. The authors discuss the implications of the results for the PCL-R and for female psychopathy more generally. PMID- 11727954 TI - MCMI-II personality traits and symptom traits in parents of children with selective mutism: a case-control study. AB - The etiology of selective mutism (SM) in children is unknown but is regarded as multifactorial. SM is frequently associated with social anxiety and language disorder or delay. The present study addresses SM and social anxiety as a familial phenomenon by comparing self-reported personality traits and symptom traits (Millon Clinical Multiaxial Inventory) in parents of 50 SM children with control parents. Personality and symptom traits reflecting social anxiety are also explored separately in the parents of SM children with and without a comorbid communication disorder. The results confirm SM and social anxiety as a familial phenomenon. Assessment and treatment planning should take this into account. The family data also suggest different family transmissions of SM and social anxiety in the SM group with and without a comorbid communication disorder. PMID- 11727955 TI - Impact of comorbid depressive symptoms on neuropsychological performance in obsessive-compulsive disorder. AB - There is indirect evidence from previous research that several executive disturbances in obsessive-compulsive disorder (OCD) are mediated by comorbid depressive symptoms. For the present study, the authors investigated whether OCD patients with elevated Hamilton Rating Scale for Depression (HRSD) scores would exhibit deficits in tasks sensitive to the medial and dorsolateral frontal cortex as well as other executive tasks. The 36 OCD patients were split along the median according to their HRSD scores and compared with matched control subjects. Patients with high HRSD scores performed significantly worse than control subjects and patients with low HRSD scores on the Wisconsin Card Sorting Test, the Trail-Making Test (TMT, Part B), and the TMT difference score. Moreover, patients with high HRSD scores exhibited deficits on a (creative) verbal fluency task. It is suggested that comorbid depressive symptoms may have artificially inflated some executive deficit scores in previous studies. PMID- 11727956 TI - Apoptosis inhibition mediated by medroxyprogesterone acetate treatment of breast cancer cell lines. AB - Several reports suggested that steroidogenic hormones could be directly involved in the regulation of apoptosis in vitro, but whether this is due to blocking or promoting mechanism of these hormones remains controversial. However, it was shown that progesterone exhibited a protective effect against the apoptotic process during mouse mammary gland involution in vivo. In this study, we analyzed the effect of medroxyprogesterone acetate (MPA) treatment, an agonist of progesterone, on serum starvation induced apoptosis on breast cancer cell lines. Positive and negative progesterone receptor (PgR+ and PgR-) breast cancer cell lines were treated with MPA (10 nM), either in standard culture conditions or in serum-free medium to induce apoptosis. Cell survival, proliferation and apoptosis were simultaneously analyzed with the expression of apoptosis-related genes measured by a real time quantitative RT-PCR. At non cytotoxic doses, MPA protected PgR+ T47-D, MCF-7 and H466-B cell lines against serum depletion-induced apoptosis, while MPA did not protect PgR-MDA-MB-231 cells against serum depletion induced apoptosis. In PgR+ cell lines and in concordance with the protective effect, the pro-apoptotic HRK and BAK1 mRNAs were up-regulated after apoptosis induction, while they were no more induced in condition of protection against apoptosis after MPA treatment. We also observed, specifically in PgR+ cells, an up-regulation of BCLX-L and BCLX-S and a down-regulation of BCL2 mRNAs, which are specific to the MPA response and unrelated to apoptotic process. Involvement of these genes with regard to the MPA-mediated protection against apoptosis is discussed. PMID- 11727957 TI - Pulmonary complications following different radiotherapy techniques for breast cancer, and the association to irradiated lung volume and dose. AB - PURPOSE: This study investigates the incidence of short-term pulmonary complications following radiotherapy (RT) for breast cancer (BC) with different treatment techniques/incidentally irradiated lung volumes and the importance of confounding factors on RT-induced pulmonary complications. PATIENTS AND METHODS: Prospectively, 475 patients with BC were followed for pulmonary complications 1, 4 and 7 months post-RT. Mean lung dose volume histograms (MDVH) were constructed and compared for the different RT-techniques. Among a subset of the mastectomized patients treated with loco-regional (LR-) RT, who had undergone complete three dimensional (3-D) dose planning (n = 43), MDVH for asymptomatic patients was compared with MDVH for patients experiencing both radiological and clinical pulmonary side-effects. RESULTS: Moderate pulmonary complications, that is requiring treatment with corticosteroids, were rare following local RT (< 1%), but were diagnosed among 11% of the patients treated with LR-RT. A correlation between increasing irradiated lung volumes at the >20 Gy-level (V20), based on MDVH for the RT-techniques, and pulmonary complications was found (P < 0.001). Furthermore, increasing age and reduced pre-RT functional level were independently associated with a higher rate of pulmonary complications (P = 0.005 and P = 0.018). Among the subgroup of mastectomized patients treated with LR-RT, who had undergone complete 3-D dose planning, a difference in mean V20 was found between patients experiencing both clinical and radiological pulmonary side effects compared to patients experiencing neither of the two side-effects (P = 0.007). CONCLUSION: Moderate pulmonary complications following local RT for BC are rare. The incidence of short-term moderate pulmonary complications in LR-RT is, however, clinically significant and to define quality assurance guidelines for these RT-techniques, 3-D RT planning can be used. PMID- 11727958 TI - Altered expression and localization of PKC eta in human breast tumors. AB - Protein kinase C (PKC) eta is a PKC isoform whose upregulation is associated with differentiation in many epithelial tissues, including the rat mammary gland. The purpose of this study was to examine whether PKC eta is altered, in expression or localization, in human breast cancer. Paraffin sections of 49 in situ breast lesions, 29 invasive breast tumors, and nine normal breast biopsies were examined for PKC eta expression by immunohistochemistry. Adjacent regions of normal epithelium, and in situ lesions that were present adjacent to invasive lesions were also analyzed. In normal epithelium, regardless of the presence of adjacent in situ or invasive lesions, PKC eta was present in the cytoplasm of the luminal epithelium, and increased in areas of normal lobular development, similar to normal rat mammary gland. PKC eta staining intensity was homogeneous in normal lobules, but heterogeneous in in situ and invasive lesions, being focally increased in cells with aberrant nuclear morphology. In situ lesions were similar to adjacent normal epithelium in average staining intensity, regardless of whether invasion was also present. However, the invasive lesions themselves were significantly decreased in staining intensity compared to adjacent in situ lesions. In addition, 75% of invasive breast cancer lesions showed decreased staining relative to adjacent normal epithelium, compared to 37% of in situ lesions. The invasive tumors which possessed high PKC eta staining were associated with positive lymph node status. These results demonstrate that quantitative and qualitative alterations in PKC eta occur in human breast cancers. PMID- 11727959 TI - Increased stromal expression of murine urokinase plasminogen activator in a human breast cancer xenograft model following treatment with the matrix metalloprotease inhibitor, batimastat. AB - The matrix metalloprotease (MMP) family of enzymes and the urokinase plasminogen activator (uPA) pathway have both been implicated in tumor invasion and metastasis and in poor prognosis of cancer. We have previously shown that treatment with batimastat, a synthetic MMP inhibitor, leads to significant retardation but not regression of tumor growth in a human breast cancer xenograft model. In addition, batimastat treatment did not inhibit local tumor invasion, nor did it encourage stromal encapsulation of the tumor, suggesting the additional involvement of non-MMP proteolytic mechanisms. To investigate the presence of an alternative extracellular matrix protease whose activity is known to be important in breast cancer, but which is not inhibited by batimastat, expression of murine and human uPA were examined by in situ hybridization and ELISA. No differences were observed between untreated and batimastat-treated tumors regarding human uPA mRNA and protein. In contrast, murine uPA mRNA expression was increased at the tumor-stromal junction in batimastat-treated tumors in comparison with the control tumors. In agreement with these results, batimastat treatment was shown to significantly induce murine uPA protein content in the tumors. Inoculating MDA435/LCC-6 cells into immunodeficient, uPA-deficient mice resulted in tumor growth retardation as compared to tumor growth in littermate wild-type controls, while addition of batimastat treatment to uPA-/- mice did not result in further growth inhibition. The increased expression of stromal uPA may represent a cellular response to MMP inhibition and may demonstrate a new level of plasticity in the malignant progression of the disease. These results may have important implications for the clinical applications of MMP inhibitors, as well as for development of other anti-invasion drugs. PMID- 11727960 TI - Predictors of distant metastasis at presentation in breast cancer: a study also evaluating associations among common biological indicators. AB - BACKGROUND: To investigate the correlation among some of the commonly used clinical, pathological factors and newer biological indicators, and to identify the independent predictors of distant metastasis at presentation in patients with breast cancer. METHODS: The pathological specimens from 73 patients with breast cancer were retrospectively evaluated by immunohistochemistry. Data on 13 biological indicators; ER, PR, P53, c-erbB-2, PCNA, CEA, Ki-67, Vimentin, Ulex, Nm23, Cathepsin D, Factor VIII, PS2 together with clinical and pathological factors were collected. RESULTS: A number of highly significant correlations were found among the biological indicators studied. By logistic regression analysis, the predictors of distant metastasis at presentation in univariate tests were tumor diameter, number of lymph nodes involved, P53, c-erbB-2 and grade. In multivariate analysis, tumor diameter (P = 0.042, HR: 1.88(1.02-3.44)), c-erbB-2 expression (P=0.035, HR: 18.20 (1.23-268.66)) and grade (P=0.010, HR: 8.05(1.66 39.00)) retained their significance. CONCLUSION: Our findings show that inactivation of suppressor genes, expression of oncogenes, loss of differentiation, augmentation of proliferative activity, metastatic potential, angiogenesis and hormone receptor status are all interrelated facets of breast cancer pathogenesis. Patients with tumors overexpressing c-erbB-2 or with bigger or higher-grade tumors probably need to be more carefully evaluated for the presence of distant metastasis, thus be better staged, at presentation. This may be a new reason to test c-erbB-2 routinely in all patients with breast cancer in addition to its well-known prognostic and predictive uses. PMID- 11727961 TI - Clinical relevance of biologic factors in male breast cancer. AB - There is ample information on the clinical role of biologic factors in female breast cancer: urokinase-type plasminogen activator (uPA), its receptor uPAR, its inhibitors PAI-1 and PAI-2, cathepsin D and pS2-protein. However such reports are missing or very rare for male breast cancer. We determined the cytosolic levels of oestrogen receptor (ER), progesterone receptor (PgR), cathepsin D, pS2 protein, uPA, uPAR, PAI-1 and PAI-2 of the primary tumour tissues from 40 male breast cancer patients. The tumour levels were compared with those of 180 matched females and 4114 historic females with breast cancer. In male breast tumours the level of PgR was higher, those of uPA, PAI-1, PAI-2 and cathepsin D lower. The tumour level of ER in men was similar to those in the matched and postmenopausal women, but much higher than those in the historic women. Male breast cancer seems to be biologically different from female breast cancer. Correlation of the eight cell biologic factors with disease outcome showed that PAI-1 (p = 0.03) was the only independent predictive factor for poor prognosis in male breast cancer. PMID- 11727962 TI - Does breast cancer grade worsen with time? Evidence from breast screening. AB - It has been suggested that alteration of the distribution of histological grades, that has been found in screening programmes, is evidence for progression in histological grade with increasing size. A predictive model, that was based upon estimated growth data from an unselected series of 98 new primary breast cancers, is used to estimate the proportions of tumors that would be diagnosed according to limiting screen diagnostic and clinical diagnostic sizes after particular screening intervals; windows of opportunity are created. The results show that the limitations imposed by time and size criteria alter the distribution of growth rates of tumours that may appear in the windows. Small screen diagnostic sizes and short screening intervals allow only the most rapidly growing tumours to reach large sizes. This produces an apparent association of grade 3 tumours with large size. Interval cancers are also likely to be more rapidly growing while the more slowly growing tumours will be diagnosed at the subsequent screen to produce a spurious association of slowly growing grade 1 tumours with small size. We conclude that the evidence from screening does not support the thesis of progression of histological grade with the ageing of the tumour, since the changes that have been observed are predictable from a simple model based upon patterns of tumour growth rates and the relationships between growth rate and histology. PMID- 11727963 TI - The validation of a quality of life scale to assess the impact of arm morbidity in breast cancer patients post-operatively. AB - This paper documents the validation of a quality of life scale (QOL) designed to assess the impact of arm morbidity on patients following breast cancer surgery. A four item arm subscale was developed to supplement a multi-dimensional, validated breast cancer QOL tool, the functional assessment of cancer therapy (FACT-B.) The new questionnaire, the FACT-B + 4, was validated on 279 women participating in a trial of sentinel node guided axillary therapy and 29 women attending a lymphoedema clinic. The subscale demonstrated good internal consistency (alpha co efficient = 0.62 to 0.88) and stability (test-retest reliability = 0.97). Lymphoedema patients reported significantly greater arm problems than a matched sample of pre-operative trial participants. The lymphoedema group also scored lower than trial patients on the FACT-B + 4 indicating a poorer quality of life (p < 0.05). A subset of 66 trial patients who had completed three consecutive assessments was used to evaluate the sensitivity of the questionnaire to change over time. Scores on the FACT-B + 4 were found to decline significantly between the pre-operative assessment and post-operative assessment at 1 month. Arm problems significantly increased during this period. FACT-B + 4 score increased again from 1 month to 12 weeks post-surgery and symptoms reduced, as the extent of arm morbidity resolved. The FACT-B + 4 appears to be psychometrically robust and sensitive to patient rehabilitation, making it suitable for use in longitudinal surgical trials. Given the dearth of existing scales available to measure arm morbidity, we hope this new tool will prove useful to researchers. PMID- 11727964 TI - Pharmacoepidemiology. PMID- 11727965 TI - Type, scope und quality of drug utilization. AB - Drug utilization research examines the quantitative and qualitative aspects of drug utilization. These include the medical, social and psychological factors and consequences of drug use in relation to specific patient groups and specific population groups, as well as the population in general (EURO-DURG-Resolution 1997). The present article puts particular emphasis on the methodological aspects and the public health perspectives involved in this search. A major goal of drug utilization research is the improvement of drug therapy as administered by physicians and pharmacists and the assessment of drug use in the population as a whole, encompassing both prescription-based and self-administer medication. In addition, drug utilization research also fulfills an important public health role in a welfare state by monitoring and controlling drug expenditure, by providing data that serve to answer health policy questions and by contributing to the management and planning of public health politics. Drug search makes use of 2 data sources. On the one hand, case-related (prescription-related) mass data (e.g. drug index) or prescriptions and medical records for patient groups held by specific institutions (e.g. IMS data, utilization observations), on the other hand, personal but anonymous prescription information and medical records taken from unselected natural populations (e.g. survey data from statutory health insurance institutions). Comparing drug utilization data for population groups in various regions, both national and international as well as between medical institutions, sociodemographic groups and diagnostic profiles will help to provide information on the medical, economical and social determinants of drug use and provide a base for health policies. PMID- 11727966 TI - Compliance with prescription recommendations by physicians in practices. AB - OBJECTIVES: Few investigations are available on the primary supply of patients in Germany. But they play a key role due to their relevance in health politics and economics. An analysis has been carried out to determine the extent to which physicians in practices adhere to the recommendations in the package inserts and to determine the consequences of this behavior regarding the safety and efficacy of prescribed drugs. METHOD: The analysis was carried out with the IMS HEALTH mediplus patient database, which is a database providing anonymous access to a representative panel of physicians and patients in Germany. More than 1,000 medical practices including more than 5 million patients and over 75 million prescriptions can be analyzed in a cross and/or longitudinal section. The evaluation time period per patient currently exceeds 10 years with monthly updates. RESULTS: Kava-Kava drug prescriptions indicate that the recommended daily dose was significantly overstepped in 78% of the cases. The recommended daily dose complied in only 41% of the cases whereas in 3 9% of the cases, the drug was underdosed. DISCUSSION: These results may give an explanation why Kava Kava prescriptions are associated with a high incidence of adverse drug reactions. On the other hand, lack of efficacy in many patients was mainly due to the fact that the drug was underdosed. PMID- 11727967 TI - Unused drugs returned to the pharmacy--new data. AB - INTRODUCTION: Although the increasing costs in the health system in Germany is a frequently discussed topic, the amount of the so-called unused drugs which patients and customers bring back to pharmacies does not seem to diminish. In 1988, the authors conducted a study to document this problem. Ten years later, in 1998, a second study with the same design was done. The new data are compared with the results of the first study. METHODS: The study was carried out in a public pharmacy in Berlin, Germany, 1998. All drugs returned unused were documented during a period of 12 months. The drugs were counted and classified according to therapeutic groups and to prescription or OTC status. The remaining amount in each package was determined in relation to the package size. The prices of the drugs were obtained from the "Rote Liste 1997". RESULTS: 10,603 unused drug-packages were collected (1988: n = 5,164). The 10 most frequent indication groups showed nearly the same ranking as the German annual report of prescribed drugs (GKV-Index 1988, same result). Only 17% of the returned drugs were for self medication (1988: 12%). The value of the original medicines amounted to approximately DM 232,920 (1988: DM 100,000), therefore, the average drug price was DM 22 (1988: DM 19). On an average, packages contained 65% (1988: 70%). 24% of the drug packages returned contained the original content, i.e. were unused (1988: 31%). 39% of the returned packages contained 51-99% of the original contents (1988: 35%) and 37% contained up to 50% of the original contents (1988: 34%). CONCLUSIONS: Although patients in Germany had to pay a higher prescription charge in 1998 than in 1988, the amount of unused drugs has increased. The reasons for this non-compliant behavior have not yet been analyzed. The results of this study suggest the need for further research for the reasons of non compliance. PMID- 11727968 TI - Self-medication in Germany. AB - We introduce the legal background of self-medication with OTC-drugs in Germany with regard to the pharmacy, drug store and health food shop distribution channels and the qualifications of the sales personnel. We give an overview of the frequency of self-medication and discuss risk/benefit in a pharmacoepidemiological context. PMID- 11727969 TI - Co- and multimedication in users of ASA and vitamin E drugs in the Federal Republic of Germany. Results of the Federal Health Surveys 1984-1999. AB - OBJECTIVE: Because ASA- and vitamin E-drugs belong to the most used OTC medications and adverse drug reactions are depending on the quantity and type of comedication we describe the pattern of usage for ASA- and vitamin E-drugs along with the total drug consumption in Germany. STUDY POPULATION, MATERIAL AND METHODS: Study participants (n = 22.560) were screened in five German health surveys (1984-1999). Total drug usage was monitored with a standardized questionnaire. Blood and serum samples were examined for clinical chemistry and hematological data as well as for selected active substances of drugs. The tocopherols and salicylic acid as the main metabolite of ASA were measured by HPLC in serum samples. RESULTS: The incidence of OTC-drug usage is high in our population. The salicylic acid concentration in the serum samples depends clearly on the reason of usage and remains fairly stable for analgesic use but decreases steadily from 1984 to 1999 for use as antiplatelet drug due to formulations with lower amount of ASA/tablet. The serum concentration of alpha-tocopherol was steadily increasing in users of vitamin E-drugs as well as in no-drug-users. CONCLUSION: Because the frequency of comedication is very high, the risk of adverse drug reactions seems to be a problem that should be considered more seriously by physicians. PMID- 11727971 TI - Pattern of outpatient drug therapy in diabetics admitted to hospital--preliminary results. AB - The prevalence of outpatient treatment with various drug groups is significantly higher in diabetics than in non-diabetic control patients. In addition to the specific diabetes-related prescriptions, diabetics were more frequently treated with drugs acting on the alimentary tract or used in metabolic disorders (ATC code A), drugs used in disorders of the blood and blood-forming organs (ATC B), cardiovascular system (ATC C), musculo-skeletal system (ATC M) and nervous system (ATC N)-- resulting in markedly higher outpatient costs in this patient group. Objective of this investigation was to analyze the pre-hospital prescription pattern of diabetics and non-diabetic controls at the time of admission to hospital. METHOD: A sample survey of a total of 189 general medical and 68 surgical admissions involving diabetics and 676 and 143 non-diabetic control patients corresponding in age, sex and main diagnosis--were analyzed with regard to selected medical and demographic characteristics and pharmacotherapy. RESULTS AND DISCUSSION: The prevalence of non-diabetic drug treatment in diabetics was highest for ATC C (87.8% and 69.1%), ATC A (40.7% and 27.9%; without A10) and ATC B (39.2% and 29.4%) in internal and surgical admissions, respectively. A substantially higher prevalence was found for ATC groups B and C in diabetics and controls admitted to medical wards than in epidemiological prescription analyses. CONCLUSION: Data indicate that the need for treatment with cardiovascular drugs and drugs used to treat disorders of the blood and blood-forming organs may be associated with a higher risk of hospitalization in general medical wards. PMID- 11727970 TI - Development of indicators for assessing the quality of prescribing of lipid lowering drugs: data from the pharmacotherapeutic quality circles in Hesse, Germany. AB - OBJECTIVE: To develop indicators based on prescription analysis in order to assess adherence using guidelines and to monitor prescribing behavior. SETTING AND MATERIAL: Eleven pharmacotherapeutic circles (PTC) of the Association of Statutory Health Insurance Physicians (KV Hesse; 10 PTCs including 155 GPs as participants, mostly high prescribers; one circle with 11 moderators trained in pharmacology). These provided a total of 183,997 drug prescriptions involving 54,970 patients (prescriptions reimbursed by the substitute fund--Ersatzkassen- II. quarter 2000); claims form from 151 GPs. On average, 5.1% of the patients with prescriptions received lipid-lowering drugs. METHOD: Development and application of indicators based on the guideline for the treatment of hypercholesterolemia developed by a GP's guideline group of the quality circles in Hesse (Hausarztliche Leitliniengruppe Hessen). The ratio of prescribing for primary and secondary prevention was chosen as a top indicator for measuring adherence to the guideline. Prescribing for secondary prevention was assessed by a set of special diagnoses. The second indicator relates to patient groups (here: older than 75 years) where the benefit of prescribing is not clear. Further indicators measure the prescribing of first choice drugs, avoidance of risk combinations and costconscious prescribing. The indicators are presented in the prescription analysis and will be discussed during the circle meeting. RESULTS: On average, the moderators prescribed 34% of the lipid-lowering drugs for primary prevention, the GP circle participants 36.7%. On average, moderators and GP participants prescribed lipid-lowering drugs for 4.9% and 7.5% of patients older than 75 years, respectively (6% and 22% in primary prevention). Approximately, 28% of all lipid-lowering drugs issued by the participants involved simvastatin and pravastatin as first-choice drugs compared to 36.5% in the case of the moderators. The prescribing of statins with fibrates or macrolides in combination is seldom. Cost-conscious prescribing can be assessed for each GP by the percentage of generic prescribing and the number of different brands per active agent. Both, moderators and participants used generics when prescribing fibrates, bile acid sequestrants or other types of lipid-lowering drugs (moderators 53.8% and GP participants 78.5%). Three months is too short a period for assessing compliance of lipid-lowering drug prescribing. CONCLUSION: It is possible to derive indicators from the guidelines and to integrate them into prescription analysis. The indicators reveal prescribing problems. The evaluation of PTCs in 2002 will involve the use of indicators as an instrument to assess the success in the implementation of guidelines. PMID- 11727972 TI - Opinion on the use of the antitumor drug trastuzumab (Herceptin) in patients with metastatic breast cancer in the county Mecklenburg-Vorpommern. AB - The recombinant humanized anti-HER-2 monoclonal antibody trastuzumab (Herceptin) is directed against the human epidermal growth factor receptor on the surface of breast cancer cells. Herceptin was approved in Germany in September 2000 after evaluation in clinical trials involving women with metastatic breast cancer who had tumors overexpressing HER-2/neu. A prerequisite for its use is the diagnosis of the HER-2/neu receptor status in individual patients because trastuzumab is only effective in patients with a high (score +3) overexpression of the HER-2/neu receptor. The only approved diagnostic method is the immunohistochemical DAKO Hercep test. Clinical experience with this novel biological agent has been obtained in 2 Phase III trials involving 469 and 222 patients, where trastuzumab was used as first- or second-line therapy. The addition of trastuzumab to chemotherapy regimens was associated with longer time to progression, a higher rate and duration of response and longer survival. When used as a single agent in metastatic breast cancer that had progressed after chemotherapy, there was an overall response rate of 15%. The median duration of response was 9.1 months and median survival was 13 months. Unwanted effects included potentially severe cardiotoxicity and in 40% of patients infusion-associated fever and/or shivering that usually occurred only during the first infusion. In patients with moderate HER-2/neu expression, unwanted drug effects outweigh a relatively weak therapeutic effect. In cases of high overexpression, the cancer may go into regression and survival may be prolonged with a relatively small impairment in the life quality. The costs of trastuzumab-therapy are high amounting to an additional 48,000 DM per patient per year. Recommendations for diagnosis and therapy in Mecklenburg-Vorpommern have been formulated in discussions between oncologists, practitioners, scientists and regulatory authorities. PMID- 11727973 TI - Drug use and adverse event monitoring in German children. AB - OBJECTIVE: To assess drug use and the incidence of mild to moderate adverse drug effects (AE) in children aged 0 - 16 years, with 2 different approaches. METHODS: All 185 pediatric practices in the State of Brandenburg, Germany, were surveyed (95% positive response). A random sample of 50 practices servicing 36,000 children participated in the study as sentinel physicians reporting any AE among their patients. Twenty practices servicing 14,000 children were included in a drug utilization component (DUC) which documented diagnoses, prescribing and AE in all children seen in a 3-month period. RESULTS: The results on the DUC are based on 12,628 children with 26,168 physician contacts, 34,465 diagnoses and 33,140 drug prescriptions given to 85% of the children seen (average age: 6.7 years). Practice size ranged from 94 to 1,473 children seen in a 3-month period. The average number of physician contacts per child was 2. 1, with 10% seeing a physician 5 times and more. The diagnoses were respiratory ailments (30%), skin- (12%), injection- (11%) and trauma- (10%) related diagnoses. Major drug uses were respiratory (34% of prescriptions), anti-infective (22%), dermatological (13%), gastrointestinal (9%) and psychotropic agents (8%). One hundred and fifty-two AE affecting 150 children were documented in the DUC, 60% of these occurring within 3 days after treatment initiation. 83% of AE were mild (up to 4 on a scale from I -6), and 85% were considered by physicians to be attributable. Only 4 reports of very severe events were received. 47% of reports were related to antibiotic uses, 31% to immunizations, 11% to respiratory, and 4% to gastrointestinal agents. While there are no differences in the occurrence of AE by gender, AE decreases significantly by age, maintaining the same patterns of AE type and causative medication. Over a 3-month period, the incidence of mild to moderate ADR in children given medications is estimated as 1,200 per 100,000, or about 1%. CONCLUSIONS: Underreporting of AE remains an issue even when a monitoring scheme suggests compliance. The results of this study show that while AE in children are generally non-serious, 1% of treatments might elicit mild AE leading to a consultation. Direct practice recording produces more reports than a sentinel system. PMID- 11727974 TI - Simultaneous measurement of intra- and intermolecular NOEs in differentially labeled protein-ligand complexes. AB - A new NOE strategy is presented that allows the simultaneous observation of intermolecular and intramolecular NOEs between an unlabeled ligand and a 13C,15N labeled protein. The method uses an adiabatic 13C inversion pulse optimized to an empirically observed relationship between 1 J(CH) and carbon chemical shift to selectively invert the protein protons (attached to 13C). Two NOESY data sets are recorded where the intermolecular and intramolecular NOESY cross peaks have either equal or opposite signs, respectively. Addition and subtraction yield two NOESY spectra which contain either NOEs within the labeled protein (or unlabeled ligand) or along the binding interface. The method is demonstrated with an application to the B12-binding subunit of Glutamate Mutase from Clostridium tetanomorphum complexed with the B12-nucleotide loop moiety of the natural cofactor adenosylcobalamin (Coenzyme B12). PMID- 11727975 TI - Measurement of 2J(H,C)- and 3J(H,C)-coupling constants by alpha/beta selective HC(C)H-TOCSY. AB - A new heteronuclear NMR pulse sequence for the measurement of nJ(C,H) coupling constants, the alpha/beta selective HC(C)H-TOCSY, is described. It is shown that the S3E element (Meissner et al., 1997a,b) can be used to obtain spin state selective coherence transfer in molecules, in which adjacent CH moieties are labeled with 13C. Application of the alpha/beta selective HC(C)H-TOCSY to a 10 nt RNA tetraloop 5'-CGCUUUUGCG-3', in which the four uridine residues are 13C labeled in the sugar moiety, allowed measurement of two bond and three bond J(C,H) coupling constants, which provide additional restraints to characterize the sugar ring conformation of RNA in cases of conformational averaging. PMID- 11727976 TI - Determining binding sites in protein-nucleic acid complexes by cross-saturation. AB - Cross-saturation experiments have been shown to give accurate information regarding the interacting surfaces in protein-protein and protein-RNA complexes. The rate of magnetization transfer depends on a number of factors including geometry, spin topology, and effective correlation times. To assess the influence of these variables on such experiments, and to determine the range of applicability of the technique, we have simulated the time-course of magnetization transfer across the interface in a variety of protein-nucleic acid complexes (434 Cro, SRY, MetJ and U1A), each having a different binding geometry. The simulations have been carried out primarily to provide information about the experimentally accessible targets for selective saturation, such as the anomeric protons and the imino protons of the nucleic acid. Saturation of either of these groups of signals leads to partial excitation throughout the nucleic acid molecule, and the resulting transfer of saturation to the labelled protein moiety can be readily detected by the reduction in intensity of particular peaks in the HSQC spectrum of the protein. The simulations show that information can be obtained about the residues in contact with the nucleic acid without any need for deuteration. Experimental cross-saturation data have been obtained from the Mbp1 DNA complex and interpreted in conjunction with the simulations to map out the binding surface in detail. PMID- 11727977 TI - Controlling residual dipolar couplings in high-resolution NMR of proteins by strain induced alignment in a gel. AB - Water-soluble biological macromolecules can be weakly aligned by dissolution in a strained, hydrated gel such as cross-linked polyacrylamide, an effect termed 'strain-induced alignment in a gel' (SAG). SAG induces nonzero nuclear magnetic dipole-dipole couplings that can be measured in high-resolution NMR spectra and used as structural constraints. The dependence of experimental 15N-1H dipolar couplings extracted from two-dimensional heteronuclear single quantum coherence (HSQC) spectra on several properties of compressed polyacrylamide, including the extent of compression, the polyacrylamide concentration, and the cross-link density, is reported for the B1 immunoglobulin binding domain of streptococcal protein G (protein G/B1, 57 residues). It is shown that the magnitude of macromolecular alignment can be widely varied by adjusting these properties, although the orientation and asymmetry of the alignment tensor are not affected significantly. The dependence of the 15N relaxation times T1 and T2 of protein G/B1 on polyacrylamide concentration are also reported. In addition, the results of 15N relaxation and HSQC experiments on the RNA binding domain of prokaryotic protein S4 from Bacillus stearothermophilus (S4 delta41, residues 43-200) in a compressed polyacrylamide gel are presented. These results demonstrate the applicability of SAG to proteins of higher molecular weight and greater complexity. A modified in-phase/anti-phase (IPAP) HSQC technique is described that suppresses natural-abundance 15N background signals from amide groups in polyacrylamide, resulting in cleaner HSQC spectra in SAG experiments. The mechanism of protein alignment in strained polyacrylamide gels is contrasted with that in liquid crystalline media. PMID- 11727978 TI - Measurement of small scalar and dipolar couplings in purine and pyrimidine bases. AB - A suite of spin-state-selective excitation (S3E) NMR experiments for the measurements of small one-bond (13C-13C, 15N-13C) and two-bond (1H-13C, 1H-15N) coupling constants in 13C,15N labeled purine and pyrimidine bases is presented. The incorporation of band-selective shaped pulses, elimination of the cross talk between alpha and beta sub-spectra, and accuracy and precision of the proposed approach are discussed. Merits of using S3E rather than alpha/beta-half-filter are demonstrated using results obtained on isotopically labeled DNA oligonucleotides. PMID- 11727979 TI - Improved TROSY-HNCA experiment with suppression of conformational exchange induced relaxation. AB - A general method for improving of the sensitivity of the TROSY-type triple resonance experiments in the presence of conformational exchange-induced (CSX) relaxation is proposed based on the use of CPMG-INEPT (Muller et al., J. Am. Chem. Soc., 1995, 117, 11043-11048) during the N-C polarization transfer periods. Significantly improved sensitivity is demonstrated for the majority of cross peaks in the new [15N,1H]-TROSY-XY-HNCA experiment, measured with partially folded RNase AS-Protein, with negligible loss of sensitivity for resonances unaffected by CSX relaxation. In addition, a comparison of cross-peak amplitudes in [15N,1N]-TROSY-XY-HNCA and conventional [15N,1H]-TROSY-HNCA spectra provides a quick and sensitive estimation of the CSX relaxation contribution. PMID- 11727980 TI - Optimized labeling of 13CHD2 methyl isotopomers in perdeuterated proteins: potential advantages for 13C relaxation studies of methyl dynamics of larger proteins. AB - 13CHD2 methyl isotopomers are particularly useful to study methyl dynamics in proteins because, as compared with other methyl isotopomers, the 13C relaxation mechanism for this isotopomer is straightforward. However, in the case of proteins, where (omega tau)2 >> 1, the refocused INEPT pulse sequence does not completely suppress unwanted 13CH3 signals. The presence of weak 13CH3 peaks is usually not a serious problem for smaller proteins because there are relatively few methyl signals and they are sharp; however, signal overlap becomes more common as the size of the protein increases. We overcome this problem by preparing a protein using a 98% D2O cell culture medium containing 3-(13)C pyruvic acid, 50-60% deuterated at the 3-position, and 4-(13)C 2-ketobutyric acid, 98% and 62% deuterated at the 3- and 4-positions, respectively. This approach significantly reduces the population of the CH3 isotopomer while optimizing the production of 13CHD2, the isotopomer desired for 13C relaxation measurements. In larger proteins where the deuterium T2 may be too short to measure accurately, we also suggest the alternative measurement of the proton T2 of the 13CH2D methyl isotopomer, because these protons are well-isolated from other protons in these highly deuterated samples. PMID- 11727982 TI - 1H, 15N, and 13C resonance assignments for a 20 kDa DNA polymerase from African swine fever virus. PMID- 11727981 TI - Pressure-stability of phospholipid bicelles: measurement of residual dipolar couplings under extreme conditions. AB - High-pressure NMR of proteins in solutions currently gains increasing interest. 3D structure determination of proteins under high pressure is, however, so far impossible due to the lack of NOE information. Residual dipolar couplings induced by the addition of magnetically orienting media are known to be capable of replacing NOE information to a very high extent. In the present contribution we study the pressure-stability of dimyristoylphosphatidylcholine (DMPC)/dihexanoylphosphatidylcholine (DHPC) bicelles and demonstrate the feasibility of measuring residual dipolar couplings in proteins under high pressure. PMID- 11727983 TI - Sequence-specific 1H, 13C and 15N resonance assignments of the N-terminal, 135 residue domain of KaiA, a clock protein from Synechococcus elongatus. PMID- 11727984 TI - 1H, 13C and 15N chemical shift assignment of the honeybee odorant-binding protein ASP2. PMID- 11727985 TI - Sequence-specific 1H, 13C and 15N resonance assignments of human GABA receptor associated protein. PMID- 11727986 TI - Backbone 1H, 13C, and 15N resonance assignments for a 14 kD protein, GABA(A) receptor associated protein (GABARAP). PMID- 11727987 TI - 1H, 13C and 15N resonance assignments and secondary structure of ADR6 DNA-binding domain. PMID- 11727988 TI - 1H, 15N and 13C resonance assignments for the gallium protoporphyrin IX-HasA(sm) hemophore complex. PMID- 11727989 TI - Paramagnetism-based versus classical constraints: an analysis of the solution structure of Ca Ln calbindin D9k. AB - The relative importance of paramagnetism-based constraints (i.e. pseudocontact shifts, residual dipolar couplings and nuclear relaxation enhancements) with respect to classical constraints in solution structure determinations of paramagnetic metalloproteins has been addressed. The protein selected for the study is a calcium binding protein, calbindin D9k, in which one of the two calcium ions is substituted with cerium(III). From 1823 NOEs, 191 dihedral angles, 15 hydrogen bonds, 769 pseudocontact shifts, 64 orientational constraints, 26 longitudinal relaxation rates, plus 969 pseudocontact shifts from other lanthanides, a final family with backbone r.m.s.d. from the average of 0.25 A was obtained. Then, several families of structures were generated either by removing subsets of paramagnetism-based constraints or by removing increasing numbers of NOEs. The results show the relative importance of the various paramagnetism-based constraints and their good complementarity with the diamagnetic ones. Although a resolved structure cannot be obtained with paramagnetism-based constraints only, it is shown that a reasonably well resolved backbone fold can be safely obtained by retaining as few as 29 randomly chosen long-range NOEs using the standard version of the program PSEUDYANA. PMID- 11727990 TI - Enhancement of the steady-state magnetization in TROSY experiments. AB - Under the condition that the longitudinal relaxation time of spin I is shorter than the longitudinal relaxation time of spin S the steady-state magnetization in [S,I]-TROSY-type experiments can be enhanced by intermediate storage of a part of the steady-state magnetization of spin I on spin S with a pulse sequence element during the relaxation delay. It is demonstrated with samples ranging in size from the 1 kDa cyclosporin to the 110 kDa 15N,2H-labeled dihydroneopterin Aldolase that intermediate storage of steady-state magnetization in a [15N,1H]-TROSY experiment yields a signal gain of 10-25%. The method proposed here for intermediate storage of steady-state magnetization can be implemented in any [15N,1H]-TROSY-type experiments. PMID- 11727991 TI - Yeast--a panacea for the structure-function analysis of membrane proteins? AB - In recent years, the scientific community has begun to realise that the structure function analysis of membrane proteins has lagged considerably behind that of their soluble counterparts. A boom in the field of membrane protein biology has resulted in the tailoring of techniques for the cloning, expression, purification and characterisation of these somewhat intractable proteins and most notably in the optimisation of several alternative host systems for this purpose. This Review Article summarises the use of yeast as a host. Compared with other hosts, it is clear that yeast combines the advantages of eukaryotes with the ease of handling of prokaryotes. Moreover, this organism provides membrane protein biologists with a panacea for structure-function analyses, not least because the tools of yeast genetics are at their disposal. PMID- 11727992 TI - Regulation of the AEFG1 gene, a mitochondrial elongation factor G from the dimorphic yeast Arxula adeninivorans LS3. AB - Oxygen influences the synthesis of mitochondrial proteins by alteration of the expression of mitochondrial genes and several nuclear genes. One of the genes localised in the nucleus is the EFG1 gene that encodes the mitochondrial elongation factor G (MEF-G). This unique gene (AEFG1) has been isolated from the non-conventional dimorphic yeast, Arxula adeninivorans LS3. The AEFG1 gene comprises a ORF of 2,274 bp, which corresponds to 757 amino acids. In the present study, the regulation of AEFG1 has been analysed for different morphological stages of A. adeninivorans and various culture conditions. It was demonstrated that the transfer of aerobically growing cultures to anaerobic conditions resulted in an accumulation of AEFG1 transcript, correlating with an increase in AMEF-G protein concentration. Since this regulation occurred in budding-cell culture growing at 30 degrees C and in both of the mycelial cultures grown at 45 degrees C and 30 degrees C, respectively, it was the oxygen level (but not the cultivation temperature or the morphological stage) which influenced the AEFG1 regulation. PMID- 11727993 TI - Hideaway, a repeated element from Ascobolus immersus, is rDNA-associated and may resemble a retrotransposon. AB - A lambda library of Ascobolus immersus stock 50 was screened for repeated DNA sequences by employing differential hybridization to identify transposable elements. Several lambda clones carrying repeated DNA sequences were obtained. One clone contains a repeated DNA sequence with unusual characteristics. This DNA element appears to be associated with rDNA sequences. Its methylation pattern is also reminiscent of rDNA. Sequence analysis indicates that all or at least many copies of the element have integrated in the external spacer sequence separating the rDNA repeats, possibly to avoid complete inactivation by methylation. The element has been named "Hideaway". It possesses structural characteristics of class I retrotransposons, most notably the presence of long direct repeats. The data presented here are discussed with respect to the ability of repeated DNA sequences to escape host defense mechanisms. PMID- 11727994 TI - Secondary DNA structure analysis of the coding strand switch regions of five Leishmania major Friedlin chromosomes. AB - As part of the EULEISH international genome project, a region of 74,674 nucleotides from chromosome 21 of Leishmania major Friedlin was subcloned and sequenced; and 31 new coding sequences were predicted. Of particular interest was a unique coding strand switching region covering 1.6 kb of DNA; and this was subjected to further investigation. Bioinformatic analysis of this region revealed an unusually high AT composition, a lack of putative hairpins and a strong curvature of the DNA in agreement with the structural characteristics of similar regions of other Leishmania chromosomes. These observations and a comparison with the secondary DNA structure of four other Leishmania chromosomes and chromosomes of different organisms could suggest a functional role of this region in transcription and mitotic division. PMID- 11727995 TI - Complete nucleotide sequence of the linear DNA plasmid pRS224 with hairpin loops from Rhizoctonia solani and its unique transcriptional form. AB - The complete nucleotide sequence of the linear DNA plasmid (pRS224-1) from the plant-pathogenic fungus Rhizoctonia solani isolate H-16 was determined; and its unique RNA transcripts were characterized. The pRS224-1 DNA consists of 4,986 nucleotides. A computer-based study of the folding of pRS224-1 at both termini predicted hairpin-loop structures. The hairpin loops consisted of the left and right termini of 236 and 264 nucleotides, respectively, and share no sequence homology. Unique poly(A) RNAs, 4.7 kb and 7.4 kb in length and hybridizing with the pRS224 DNA, were found in mycelial cells of R. solani H-16. Transcript product-mapping allowed the prediction of the locations of different expression signals. The 7.4-kb transcript is generated from the left terminal region of the complementary strand, via the full-length sense-strand, to the right terminal region of the complementary strand. The 4.7-kb transcript is generated from the center region of the sense strand to the right terminal region of the complementary strand. One open reading frame (ORF) found in pRS224-1 is 887 amino acids long and has a potential coding capacity of 102 kDa. The ORF contains the highly conserved domains characteristic of reverse transcriptase sequences, including the highly conserved YXDD sequence. PMID- 11727996 TI - Low temperature affects the processing pattern and RNA editing status of the mitochondrial cox2 transcripts in wheat. AB - We investigated the effects of low temperature on the expression of mitochondria encoded genes in wheat. During the course of our study, we found that the transcriptional pattern of cox2 was affected by low temperature. Northern blot analysis indicated that the precursor transcript before intron splicing was clearly increased by cold, although the level of mature message was not changed. We also found that cold treatment severely suppressed RNA editing at intron binding site 1 of the precursor transcript, which plays an important role in intron splicing. These results suggest that low temperature could affect the post transcriptional regulation of wheat mitochondrial cox2 transcripts, i.e. intron splicing and/or RNA editing. PMID- 11727997 TI - Heat stress results in incomplete C-to-U editing of maize chloroplast mRNAs and correlates with changes in chloroplast transcription rate. AB - The editing status rps14 and rpl20 mRNAs decreased rapidly from nearly 100% edited to about 30% edited when maize plants were shifted from 20 degrees C to 37 degrees C. A decrease in the extent of editing was easily detected within 2 h and decreased to a steady-state level of about 40% C-to-U conversion at 37 degrees C. In contrast, the editing status of these chloroplast mRNAs increased relatively slowly after plants were shifted from 37 degrees C to 20 degrees C. Chloroplasts isolated from maize plants which were grown at 20 degrees C and then shifted to 37 degrees C for 24 h were 5-10 times more transcriptionally active than chloroplasts isolated from maize plants grown continuously at 20 degrees C. Thus, the high transcription rate at 37 degrees C may establish a kinetic condition where the rate of transcription exceeds the capacity of the editing apparatus and results in incomplete editing. PMID- 11727998 TI - Lack of conservation of editing sites in mRNAs that encode subunits of the NAD(P)H dehydrogenase complex in plastids and mitochondria of Arabidopsis thaliana. AB - RNA editing in the plastids and mitochondria of higher plants involves C to U conversion of specific nucleotides in the mRNA. This leads to the synthesis of proteins that are different from those predicted by the DNA sequence. Editing appears to have arisen at about the same time in both plastids and mitochondria, suggesting a common evolutionary origin. The problem we address here is whether or not there has been co-evolution of the editing systems in the two organelles. Our test system was editing of the Arabidopsis thaliana mRNAs for ndhB and nad2, and for ndhD and nad4, which encode homologous subunits of the plastid and mitochondrial NAD(P)H dehydrogenases, respectively. The editing sites in the Arabidopsis nad2 and nad4 mRNAs have previously been determined and we report here 19 editing sites in eight mRNAs in Arabidopsis plastids. Out of these, eight sites are localized in the ndhB mRNA. In its mitochondrial counterpart, nad2, 31 editing sites are present, none of which are shared with the ndhB gene. The Arabidopsis ndhD mRNA is edited at four positions, only one of which is shared by its mitochondrial homologue, nad4, which contains 32 editing sites. These findings suggest that, although editing in the two organelles may have derived from a single system, there is no significant conservation of editing sites in cognate mRNAs in plastids and mitochondria. PMID- 11727999 TI - Alternative medicine: herbal drugs and their critical appraisal--part II. AB - The availability of herbal products as "over-the-counter" drugs and their increasing usage in the US and Canada have caused concern at the US FDA, since these products are not currently monitored for their safety, efficacy and quality. Reliable information on these aspects of the products is not available. Most of the clinical trials carried out to date have been lacking in scientific design, data collection and interpretation, and systematic toxicological evaluation. A critical review of the existing data on three of the widely used herbs and their products is presented. Of the three herbs, garlic and ginger, through both experimental and clinical data, as well as their liberal consumption by man over millennia, appear to be very safe for therapeutic use. However, further and large-scale, well-designed clinical investigations are needed to establish their efficacy before they can enter the mainstream drug market of North America. It is hoped that this review will equip the physicians and interested biomedical scientists with a comprehensive summary of the total information available to date on the herbs described. PMID- 11728000 TI - Applications of yeast in drug discovery. AB - The yeast Saccharomyces cerevisiae is perhaps the best-studied eukaryotic organism. Its experimental tractability, combined with the remarkable conservation of gene function throughout evolution, makes yeast the ideal model genetic organism. Yeast is a non-pathogenic model of fungal pathogens used to identify antifungal targets suitable for drug development and to elucidate mechanisms of action of antifungal agents. As a model of fundamental cellular processes and metabolic pathways of the human, yeast has improved our understanding and facilitated the molecular analysis of many disease genes. The completion of the Saccharomyces genome sequence helped launch the post-genomic era, focusing on functional analyses of whole genomes. Yeast paved the way for the systematic analysis of large and complex genomes by serving as a test bed for novel experimental approaches and technologies, tools that are fast becoming the standard in drug discovery research PMID- 11728001 TI - Sympathetic nervous system and experimental diabetes: role of adrenal medullary hormones. AB - The sympathetic nervous system is of major importance in the regulation of various physiological functions. The present review discusses the mechanisms which control glucose homeostasis and the role of the sympathetic nervous system in experimental diabetes with special emphasis on the role of adrenal medullary hormones, over-activity of the sympathetic nervous system and its relationship to hypertension in the diabetic state and the effect of stress. The chapter also reviews the ability of various drugs and pharmacological agents to produce hyperglycemia in experimental animals and how this information can be used in screening new chemical entities and in differentiating the mode of action of these agents. PMID- 11728002 TI - From outer to inner space: traveling along a scientific career from astrochemistry to drug research. AB - This professional history describes my journey as a research scientist after my early training and experiences in the pre- and early post-World War II United States. My graduate training concentrated on a problem in astrochemistry: phenomena on comets. As my career developed, I felt confident enough in myself as an experimentalist to enter, and make contributions to, several different fields: structural biochemistry (via nuclear magnetic resonance spectroscopy), molecular immunology, pharmacology, neurochemistry, and cell biology. One emphasis is on the nature and quality of my scientific training that permitted me to do cross disciplinary work. A second emphasis is on the technical and intellectual developments in late twentieth-century science and how, along with the changes in American society as it passed through three major wars, they influenced my life and thought. PMID- 11728003 TI - Virus population dynamics, fitness variations and the control of viral disease: an update. AB - Viral quasispecies dynamics and variations of viral fitness are reviewed in connection with viral disease control. Emphasis is put on resistance of human immunodeficiency virus and some human DNA viruses to antiviral inhibitors. Future trends in multiple target antiviral therapy and new approaches based on virus entry into error catastrophe (extinction mutagenesis) are discussed. PMID- 11728004 TI - Dopamine fails to inhibit Na,H-exchanger in proximal tubules of obese Zucker rats. AB - Dopamine via the activation of D1-like receptors inhibits Na,K-ATPase and Na,H exchanger and subsequently increases sodium excretion. We have previously reported that dopamine failed to inhibit Na,K-ATPase in the proximal tubules (PTs) of obese Zucker rats. The present study was designed to determine the effect of dopamine on Na,H-exchanger in PTs of lean and obese Zucker rats, and examine D1-like receptor-coupled signal transduction pathway mediating the inhibition of Na,H-exchanger. We found that dopamine inhibited Na,H-exchanger in the PTs of lean rats but this response was absent in obese rats. In brush border membranes, [3H]SCH 23390 binding revealed a approximately 45% reduction in D1 like receptor binding sites in obese compared to lean rats. Dopamine stimulated cAMP accumulation in PTs of lean but not in obese rats. Forskolin-mediated stimulation of cAMP was similar in lean and obese rats. Dopamine as well as forskolin and dibutyryl cAMP-mediated stimulation of protein kinase A (PKA) was reduced in PTs of obese compared to lean rats. The data suggest that reduction in D1-like receptor binding sites, defective coupling with signaling pathway and inability of PKA activation may be responsible for the failure of dopamine to inhibit Na,H-exchanger in PTs of obese rats. This phenomenon may contribute to an increase in sodium reabsorption and development of hypertension in obese Zucker rats. PMID- 11728005 TI - Haplotypes of aldosterone synthase (CYP11B2) gene in the general population of Japan: the Ohasama study. AB - Since the identification of a chimeric aldosterone synthase which induces mendelian hypertension, polymorphisms in aldosterone synthase (CYP11B2) has been one of major targets for molecular analyses in association with hypertension. To date, four polymorphic variants of CYP11B2, -344T/C at promoter region, a gene conversion in intron 2, 2713A/G (in exon 3) which converts from Lys to Arg at codon 173 (K173R), and 4986T/C (in exon7) which converts from Val to Ala at codon 386 (V386A), have been identified in Caucasian population. Then, linkage disequilibrium between -344T/C polymorphism and a gene conversion in intron 2 or K173R mutation has been described, suggesting the presence of genetic haplotypes in Caucasians. Since the presence of a gene conversion in intron 2 or V386A mutation was still unknown in the Japanese population, all these polymorphisms were examined together to determine the CYP11B2 haplotypes of Japanese, using DNA samples from 1290 participants of the Ohasama study, who represent the general population of a rural community of northern Japan. Molecular analyses demon- strated the presence of a gene conversion of intron 2, but the absence of V386A mutation in Japanese population. The complete linkage disequilibrium between 344T/C polymorphism and K173R mutation was noted. Although -344T allele was linked either with a gene conversion in intron 2 or with normal intron 2, -344C allele was completely linked with normal intron 2. These results indicate the presence of 3 allelic haplotypes of CYP11B2, -344C with normal intron 2 and 173R, -344T with normal intron 2 and 173K, and -344T with converted intron 2 and 173K, in the general Japanese population. The frequency (total 1.0) was 0.35, 0.53, and 0.12, respectively. The presence of allelic haplotypes is considered to be an additional genetic information to individual polymorphism of CYP11B2 to determine the linkage between CYP11B2 polymorphisms and hypertension. PMID- 11728006 TI - L-NAME, a nitric oxide synthase inhibitor, as a potential countermeasure to post suspension hypotension in rats. AB - A large number of astronauts returning from spaceflight experience orthostatic hypotension. This hypotension may be due to overproduction of vasodilatory mediators, such as nitric oxide (NO) and prostaglandins. To evaluate the role of the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) as a countermeasure against the post-suspension reduction in mean arterial pressure (MAP), we assessed the cardiovascular responses and vascular reactivity to 7-day 30 degrees tail-suspension and a subsequent 6 hr post-suspension period in conscious rats. After a pre-suspension reading, direct MAP and heart rate (HR) were measured daily and every 2 hrs post-suspension. The NO synthase inhibitor L NAME (20 mg/kg, i.v.), or saline, were administered after the 7th day reading prior to release from suspension and at 2 and 4 hrs post-suspension. At 6 hrs post-suspension, vascular reactivity was assessed. While MAP did not change during the suspension period, it was reduced post-suspension. Heart rate was not significantly altered. L-NAME administration reversed the post-suspension reduction in MAP. In addition, the baroreflex sensitivity for heart rate was modified by L-NAME. Thus, the post-suspension reduction in MAP may be due to overproduction of NO and altered baroreflex activity. PMID- 11728008 TI - Increased expression of biglycan mRNA in pressure-overloaded rat heart. AB - Biglycan mRNA expression in rat myocardium after abdominal aortic banding with renal ischemia was examined. The Northern blot analysis demonstrated that expression of biglycan mRNA in the pressure-overloaded hearts on days 2, 7, 14 and 28 was 2.88 +/- 0.89, 2.32 +/- 0.49, 2.17 +/- 0.57 and 1.81 +/- 0.46-fold higher, respectively, than that in the sham-operated hearts. In situ hybridization showed an increased density of biglycan mRNA signal-positive cells in the pressure-overloaded hearts. The cells with positive signals were spindle shaped mesenchymal cells in the myocardial interstitium. A marked increase in biglycan mRNA signal expression was also observed in endothelial cells and smooth muscle cells of the thickened myocardial capillary wall. These results demonstrated an increase in biglycan mRNA in the pressure-overloaded heart in mesenchymal cells in the myocardial interstitium, and in endothelial and smooth muscle cells of the capillaries, indicating that biglycan contributes to the ventricular and vascular remodeling in response to pressure overload. PMID- 11728007 TI - Exaggerated natriuresis after selective AT1 receptor blockade in Dahl salt sensitive rats. AB - Salt-sensitive individuals are susceptible to develop hypertension when exposed to high salt-diet. Such a phenomenon is considered to be due to a genetic impairment in the renal excretion of sodium. In the present studies extent of endogenous angiotensin-II (Ang-II) mediated antinatriuresis was comparatively evaluated in Dahl salt-sensitive (SS) and salt-resistant (SR) rats, using a selective AT1 receptor antagonist, candesartan. In addition, differences in plasma renin activity and characteristics of Ang-II receptors in the renal cortical tubular membranes were also examined. Under INACTIN anesthesia AT1 receptor blockade resulted in significant increases in renal sodium excretion, which was several-fold greater in SS rats than that observed in SR rats. These observations suggest that antinatriuretic function of endogenous angiotensin-II is exaggerated in SS rats. This functional overexpression appears to be related to an increase in the affinity of Ang-II receptors in renal cortical tubular membranes but not to receptor density or plasma renin activity. It is proposed that salt-dependent hypertension in Dahl salt-sensitive rats may be due to enhanced Ang-II mediated sodium retention. PMID- 11728009 TI - Effects of gender and age on the cardiac baroreceptor reflex in hypertension. AB - The present study examined whether alterations in the cardiac baroreceptor reflex in hypertension may be a function of constitutional differences associated with gender and age. These hypotheses were tested using a cross-sectional design that compared 20 normotensive and 21 hypertensive men and women of varying age for differences in baroreceptor reflex sensitivity and response latency for heart rate, obtained using a modified bolus phenylephrine (Oxford) method. Relative to their respective normotensive controls, baroreceptor reflex sensitivity was reduced in hypertensive men, but not in hypertensive women. Among normotensive subjects, men had greater baroreceptor reflex sensitivity than women. Independent from the effects associated with differences in blood pressure, age was not a significant predictor of reduction in baroreceptor reflex sensitivity. However, a combination of high blood pressure and older age was associated with a significant increase in baroreceptor reflex response time. In summary, gender and aging interacted with hypertension to alter two different aspects of the baroreceptor reflex. These results provide a preliminary indication that a decline in arterial baroreflex sensitivity may be more specific to hypertension in men than in women. Prolongation in baroreflex response latency in older hypertensive subjects also suggested that aging and hypertension may have a synergistic effect on cardiac parasympathetic function. PMID- 11728010 TI - Sino-aortic denervation alters the hemodynamic response to exercise in hypertensive rats. AB - The effect of sino-aortic denervation (SAD) on the heart rate (HR), arterial pressure (AP) and regional blood flow responses during dynamic exercise was examined in spontaneously hypertensive rats (SHR). Intact (n= 14) and SAD (n= 17) rats were instrumented with arterial catheters and mesenteric and iliac Doppler ultrasonic flow probes. After recovery, all rats underwent a graded exercise test. Heart rate increased significantly during exercise in intact and SAD rats. There was no significant difference in the steady state heart rate response to exercise in the intact and SAD rats. Arterial pressure increased during exercise in the intact rats. In sharp contrast, arterial pressure decreased during exercise in the SAD rats. Iliac vascular conductance increased during exercise in the intact and SAD rats. The increase in iliac vascular conductance during exercise was significantly greater in the SAD rats. Mesenteric vascular conductance decreased during exercise in the intact and SAD rats. The decrease in mesenteric vascular conductance during exercise was significantly attenuated in the SAD rats. Results suggest that functioning arterial baroreceptors are required for the typical hemodynamic responses during dynamic treadmill running in hypertensive rats. PMID- 11728011 TI - Impairment of amino-acid absorption in suckling rats infected with Cryptosporidium parvum. AB - In the present study. we explored the nutritional consequences of cryptosporidiosis. In order to ascertain the direct responsibility of C. parvum for impairment of staturoponderal development observed during the infection in neonatal animals, we investigated the absorption of two major components of the total amino acids in dam's milk (leucine and glutamate) across the ileal mucosa. The infection resulted in significant (47% and 34%, respectively) reductions in leucine and glutamate fluxes (P<0.01). Moreover, the leucine aminopeptidase and alkaline phosphatase activities were reduced in the infected ileal mucosa. Interestingly, the reduction in weight gain, which began at day 6 post-infection (PI), persisted until day 20 PI, although no cryptosporidia were detected in the ileal mucosa after day 12 PI. We thus provide evidence that the malabsorption of amino acids during cryptosporidiosis contributes to impairing the development of neonatal animals, with consequences that persist beyond eradication of the parasite. PMID- 11728012 TI - Early response gene expression during differentiation of cultured Leishmania donovani. AB - The promastigote form of the unicellular parasite, Leishmania donovani, must differentiate into the amastigote form to establish an infection in a mammalian host. Identification of genes whose expression changes during differentiation could help reveal mechanisms of Leishmania gene regulation and identify targets for controlling the diseases caused by this human pathogen. Two genomic clones were isolated, P9 that is more highly expressed in promastigotes than in axenic amastigotes and A14 that is preferentially expressed in axenic amastigotes. Analysis of the DNA sequences revealed open reading frames that would encode 55.5 kDa and 100 kDa proteins, respectively, with no homology to known proteins. The mRNA level for these genes during 24 h time courses of parasite differentiation in culture was compared to two genes known to be differentially expressed, c-lpk2 and mkk. Changes in RNA level occurred within 2 h for each gene and continued in advance of morphological changes. The expression levels of these four genes in axenic amastigotes correlated with results from animal-derived parasites. PMID- 11728013 TI - Characterisation of a monoclonal antibody recognising specifically the HSP70 from Leishmania. AB - Heat-shock protein 70 (HSP70) is ubiquitously distributed along the evolutionary scale and has such an amino acid sequence conservation that it is considered the most evolutionarily conserved protein. In order to obtain immunological tools specific against Leishmania infantum HSP70, hybridomas were established that secreted monoclonal antibodies (mAbs) against recombinant L. infantum HSP70. One of them, named mAb 2B8D2, specifically reacted with the Leishmania protein and did not recognise HSP70 from the related kinetoplastid Trypanosoma cruzi. The use of synthetic peptides allowed us to determine the B-cell epitope recognised by this mAb, an epitope located in the divergent C-terminal domain of the protein. Remarkably, the mAb possesses the capacity to immunoprecipate HSP70 from promastigote extracts of L. infantum. The fact that human HSP70 is not recognised by this mAb assures the usefulness of this antibody for diagnostic purposes and studies involving Leishmania infection of macrophages. PMID- 11728014 TI - Triazinyl derivatives that are potent inhibitors of glucose transport in Trypanosoma brucei. AB - Glucose transport in Trypanosoma brucei is facilitated by a transporter that is kinetically markedly different from its mammalian homologue. In this regard, the trypanosomal transporter may be selectively targeted. We investigated the potential of a series of triazinyl derivatives as inhibitors of glucose transport in T. brucei. A graded response of glucose transporter inhibition by these compounds was observed, with Cibacron blue 3GA, CiB, being the most potent. This inhibited transport by up to 90% in a concentration-dependent manner, with an IC50 of about 19.4 microM. A Dixon plot of different concentrations of this triazine and the rate of transport suggested that inhibition may be simple and competitive. The inhibition constant Ki was 14.8 microM. Although cytochalasin B has been widely reported to inhibit glucose uptake by mammalian and other eukaryotic glucose transporters, it had no effect at all on the trypanosome transporter at concentrations equivalent to those of the triazines. This may suggest structural differences between the trypanosome and mammalian glucose transporters and also suggests that the triazine moiety may serve as a template for the design potent trypanocides targeted at the glucose transporter. PMID- 11728015 TI - Residual infection of 15 toxoplasma strains in the brain of rats fed cysts. AB - Thirty-seven groups of 4-32 Wistar rats were 2-10(3) cysts of 15 Toxoplasma strains. After 2 months, the rats were euthanized and their brains screened for Toxoplasma cysts and bioassayed in mice if negative. The brains of 323 of 411 rats (78.6%) were found to be infected 2 months after inoculation with Toxoplasma cysts. Two hundred cysts were necessary to infect nearly 90% of the rats. With lower doses, only 60% of the rats had residual brain infection. Brain cysts were formed only in 146 of 411 rats (35.5%). The numbers of cysts formed were in the order of tens to hundreds, only occasionally one or two thousands. The mean percentage of rats with brain cysts, and the number of cysts formed in rat brains by different inocula, increased with higher doses of cysts and then declined. This pattern is difficult to explain and similar results regarding the number of cysts formed have been published. In relation to the mean percentage of rats infected, there appears to be a plateau in infection with the higher inocula. Neither the number of rats with cysts in their brains nor the numbers of cysts formed were dependent on the Toxoplasma strain used, with the exception of one strain. Instead, individual variations were marked, and are presumably related to variations in the individual genetic resistance to Toxoplasma infection in the rat. The information gathered is considered a preliminary step for a rat model of immunity against acquired toxoplasmosis. PMID- 11728016 TI - The measurement of parasite antigen-specific IgE levels using anti-IgE monoclonal antibodies and biotinylated antigens. AB - We studied and evaluated an ELISA system, using a sandwich method with a monoclonal antibody against the Fc domain of IgE molecules and biotinylated antigens, to detect parasite antigen-specific IgE quantitatively. The specific IgE ELISA titre increases linearly in a dose-dependent manner when the concentration of total IgE in samples is less than 2,000 ng/ml. Sera from IgE deficient SJA/9 mice infected with Trichinella spiralis failed to give any measurable IgE, suggesting that other classes of immunoglobulins have no effect on this assay. The titre showed a good correlation with PCA titre. A high concentration of the serum from Toxocara canis-infected mice reduced the T. spiralis-specific IgE ELISA titre, suggesting that the ELISA system is influenced by a huge amount of IgE against epitopes different from those of target antigens. This ELISA system can also be applied for detecting other classes or subclasses of antigen-specific immunoglobulins. PMID- 11728017 TI - Activity of the cyclic depsipeptide emodepside (BAY 44-4400) against larval and adult stages of nematodes in rodents and the influence on worm survival. AB - The present investigations deal with the activity of the cyclic depsipeptide emodepside (BAY 44-4400) against larval and adult stages of three rodent nematodes. While emodepside acts strongly against the adult stages of the rat nematodes Nippostrongylus brasiliensis and Strongyloides ratti, as well as against the mouse nematode Heligmosomoides polygyrus, its actions against the larval stages of these nematodes vary according to the species. Thus, emodepside is highly effective against the lung and intestine larval stages of N. brasiliensis and S. ratti. By contrast. the larval stages of H. polygyrus in the intestine are only partly affected by higher emodepside dosages. PMID- 11728018 TI - Ex vivo diffusion of albendazole and its sulfoxide metabolite into ascaris suum and Fasciola hepatica. AB - The current experiments compare the pattern of ex vivo uptake (diffusion) of albendazole (ABZ) and albendazole sulfoxide (ABZSO) by Ascaris suum and Fasciola hepatica. Specimens of A. suum and F. hepatica were collected from untreated animals (pigs and sheep, respectively) and incubated with either ABZ or ABZSO for different time periods (5-180 min). After incubation. the parasite material was analysed by HPLC to quantify the amount of ABZ and/or ABZSO. The parent drug and its active ABZSO metabolite were recovered from the parasites after ex vivo incubation for different time periods throughout the assay. Total drug availability in A. suum, expressed as area under the concentration versus time curve (AUC) over 180 min of incubation, was significantly greater (P<0.05) for ABZ parent drug (AUC = 4.19 +/- 0.59 microg x h xg(-1)) compared with the more polar ABZSO metabolite (AUC = 0.25 +/- 0.01 micro x h x g(-1)). Similar results were observed after the incubation of both molecules with F. hepatica, where the AUC values obtained were 10.6 +/- 0.28 microg x h x g(-1) and 2.04 +/- 0.33 microg x h x g(-1) for ABZ and ABZSO, respectively. The greater diffusion and availability of ABZ in both helminths correlate with the higher lipophilicity of the parent drug, compared with its sulfoxide metabolite. The amount of both molecules measured in A. suum was significantly lower (P<0.05) than that recovered in F. hepatica. The complexity of the histological structure of the nematode cuticle compared with the external tegument of the trematode may account for such a difference in drug diffusion between the species. These findings complement previous observations on the patterns of in vivo uptake of ABZ by different helminth parasites, contributing to the understanding of the pharmacological anthelmintic action of these moieties. PMID- 11728019 TI - Cell surface carbohydrates and in vivo infectivity of glucantime-sensitive and resistant Leishmania (Viannia) guyjanensis cell lines. AB - The cell surface plays an important role in the interaction of parasites with their hosts. Drug resistance in the protozoan Leishmania may involve changes in cell-surface composition, although it is not known whether infectivity is also affected. One sensitive and two glucantime-resistant lines of Leishmania (Viannia) guyanensis previously isolated were inoculated into hamsters. The sensitive line caused the disease to manifest earlier than the resistant lines. Imprinting analyses of infected macrophages showed that the sensitive line was more infective than the resistant cell lines. In vitro drug resistance was evaluated and the comparative analyses of dose-response curves showed that the susceptibility pattern of the sensitive line did not change after passage in animals, but a decrease in drug resistance was observed in resistant cell lines recovered from the mammalian host. Cell surface carbohydrates of sensitive and resistant cell lines were analysed before and after passage in animals by agglutination tests with several plant lectins. Passage in animals changed the agglutination pattern for many lectins from all three cell lines. Loss of reactivity to lectins seemed to be correlated with a decrease in infectivity of the parasite-resistant cell lines. This study opens possibilities for exploring the relationship between drug susceptibility, infectivity and surface carbohydrate composition of protozoan parasites. PMID- 11728020 TI - Some factors influencing transmission of toxoplasma in pregnant rats fed cysts. AB - An overall 44% transplacental transmission rate was observed in 221 rats fed cysts of 12 Toxoplasma strains at 15 days of pregnancy, with a range of 0-90% transmission. Considerable variability in the transmission rate was seen among different groups of rats that received similar Toxoplasma inocula; this is attributed to genetically based susceptibility to Toxoplasma among individuals of the outbred Wistar strain of rats. Transplacental transmission was more frequent in Long Evans than in Wistar rats. Significant differences in the rate of transmission were not found between rats that were fed similar Toxoplasma inocula 6-8 days or 15 days after conception. The frequency of transmission was not affected by the strain or dose of Toxoplasma used. PMID- 11728021 TI - Fasciola hepatica: the characteristics of experimental infections in Lymnaea truncatula subjected to miracidia differing in their mammalian origin. AB - Experimental infections of Lymaea truncatula, using two susceptible snail populations (Berneuil, or Migne, central France) and four isolates of Fasciola hepatica miracidia differing in their mammalian host of origin (cattle, nutrias, rabbits, or sheep), were performed under laboratory conditions to determine whether the host of origin had an effect on the daily production of cercariae. Snails were each subjected to bimiracidial exposures and were then reared under semi-natural conditions (a constant temperature of 20 degrees C and natural photoperiod). Significantly lower values were noted in the rabbit groups for survival rates at day 30 post-exposure, as well as for prevalences of infection, snail growth. duration of shedding period, and the total numbers of cercariae these snails shed. The total number of cercariae shed by both nutria groups was significantly higher than those recorded in the six other infected groups. In the cattle, rabbit, and sheep (Berneuil only) groups, the peaks in the daily distribution of cercariae occurred between day 2 and day 4 after the first shedding, and the number of cercaria-shedding snails decreased with increasing number of shedding waves. In contrast, in the three other groups, the peaks were only observed between days 20 and 45. Snails shedding their cercariae during nine or more waves were numerous in these last groups. No infradian-type rhythm in the daily distribution of cercarial numbers over the shedding period was noted for any snail group. The highest production of F. hepatica cercariae in both nutria groups would be a consequence of a higher success rate of miracidia when they infected an allopatric population of snails. The absence of an infradian-type rhythm in the distribution of daily cercarial numbers in the eight groups suggests that this rhythm, if it occurs, would only be influenced by temperature and thus be limited to periods with optimal conditions for cercarial shedding. PMID- 11728022 TI - Molecular characterization of Cryptosporidium isolates obtained from human and bovine infections in Japan. AB - Cryptosporidium oocysts, morphologically identified as Cryptosporidium parvum, were isolated from 22 human and 14 bovine cases in Japan, and were genotyped by means of a PCR/RFLP analysis of the polythreonine gene. DNA profiles of human isolates gave three distinct genotypes, namely an anthroponotic genotype 1, zoonotic genotype 2 and a new genotype. Isolates from bovine samples gave zoonotic genotype 2. The unusual genotype of Cryptosporidium was isolated from the feces of three immunologically healthy adults, and was further characterized by the sequence analysis of the 18S rRNA gene. The third genotype was identified as Crypto sporidium meleagridis, demonstrating that C. meleagridis, which occurs worldwide, has the potential to infect humans regardless of their immunological condition. PMID- 11728023 TI - A factor isolated from Taenia solium metacestodes stimulates T lymphocytes to proliferate and produce gamma interferon. AB - A metacestode factor (MF) isolated from live metacestodes of Taenia solium suppresses humoral and cellular responses, and inhibits the inflammatory reaction around metacestodes implanted subcutaneously in mice. When this MF is digested with RNase (dMF), it loses the suppressive capacity, but acquires T-cell stimulant ability. By filtering MF through a Bio-gel P6 column, two components were separated. The first (F1) was suppressive. while the second (F2) stimulated T cells to proliferate. In these experiments, F2 or dMF was used with mouse spleen cells in stimulation assays in vitro. Spleen cells from mice treated with F2 or dMF were also stimulated with concanavalin A (Con-A) ex vivo. Flow cytometry analyses were performed to estimate cell proliferation, intracellular cytokine production. and restoration of CD4 cells. Spleen lymphocytes from mice previously treated with F2 or dMF and then stimulated with Con-A ex vivo exhibited a significant increase in cell proliferation and gamma interferon production by CD4+ (P<0.05) and CD8+ cells. These effects were concentration dependent and inversely correlated with the amount of dMF or F2. Similar results were observed in normal mouse spleen T cells incubated with F2 or dMF and Con-A in vitro. Finally, dMF induced a significant restoration of CD4-cells in mice depleted of these cells. PMID- 11728024 TI - Effect of drugs inhibiting spermidine biosynthesis and metabolism on the in vitro development of Plasmodium falciparum. AB - Treatment of Plasmodium falciparum with the potent inhibitor dicyclohexylamine completely arrests in vitro cell proliferation of the chloroquine-susceptible P. falciparum strain NF54 and the R strain, which shows less sensivity to chloroquine. The average inhibitory concentration (IC50) values determined for both strains revealed different inhibition profiles. The IC50 value for the chloroquine-sensitive NF54 strain was 97 microM and 501 microM for the R strain. Monitoring polyamine pools after treatment with dicyclohexylamine leads to a significant decrease in the intracellular spermidine content, which was nearly reversed by supplementation with spermidine. Since spermidine is an important precursor for the biosynthesis of hypusine and homospermidine in eukaryotes, we studied the developmental effect on both P. falciparum strains of 1,7 diaminoheptane as an inhibitor of deoxyhypusine synthase (EC 1.1.1.249) in mammalian cells, and agmatine as a moderate inhibitor of homospermidine synthase (EC 2.5.1.44). Inhibition profiles with 1,7-diaminoheptane resulted in an IC50 value of 466 microM for the NF54 strain and 319 microM for the R strain. Spermidine pools changed significantly. Inhibition with agmatine caused a strong decrease in parasitemia for the chloroquine-susceptible NF54 strain, with a determined IC50 value of 431 microM and an IC50 value of 340 microM for the less chloroquine-susceptible R strain. Spermidine was not detectable after inhibition. The uncommon triamine homospermidine occurred in both P. falciparum strains. To our knowledge this is the first evidence of homospermidine in P. falciparum. The use of specific inhibitors of spermidine metabolism might be a novel strategy for the design of new antimalarials, and suggests the occurrence of both enzymes in the parasite. PMID- 11728025 TI - Chemotherapeutic approaches to protozoa: Coccidiae--current level of knowledge and outlook. AB - Progress in the treatment and prophylaxis of cyst-forming coccidial infections (Neospora, Sarcocystis Toxoplasma) and Cryptosporidium infections has been limited (Table 1: Haberkorn 1996: Croft 1997: Wang 1997). However, new possibilities have been opened up in the treatment of Eimeria-induced coccidiosis in poultry and mammals. due to improvements in treatment and, or metaphylaxis. A new polyether antibiotic. semduramycin, has recently been added to the range of effective prophylactic preparations. The development of resistance to anticoccidial agents is now posing similar problems to those encountered with malaria, coccidiosis in poultry being particularly affected. Because no new active ingredient from a new family of chemical substances has been developed for more than 10 years, the following approaches are being adopted to get round this problem: the use of older preparations which have not been used for a long time, the introduction of combinations such as narasin nicarbazin or methyl benzoquate clopidol and the alternating use of anticoccidial agents in rotation and shuttle programmes. The goal of a real alternative, i.e. vaccination, has been achieved to a certain extent in the form of live vaccines for laying hens and broiler breeders and is being practiced in some countries. PMID- 11728026 TI - The anthropology of nurse-anthropologists. Introduction. PMID- 11728027 TI - Betwixt and between. PMID- 11728028 TI - Anthropologist of domestic care. PMID- 11728029 TI - Current issues in using anthropology in nursing education and services. PMID- 11728030 TI - Care and culture: an introspective commentary. PMID- 11728031 TI - Notes of a nurse-anthropologist. PMID- 11728032 TI - The way of one nurse-anthropologist. PMID- 11728033 TI - Development of live recombinant vaccine candidates against tuberculosis. AB - We report on 2 recombinant (r) live vaccine candidates that use different strategies to improve cell-mediated immunity for TB control: (i) the current TB vaccine strain Mycobacterium bovis BCG, endowed with a pore-forming cytolysin; and (ii) rSalmonella typhimurium secreting a single immunodominant antigen shared by BCG and M. tuberculosis. PMID- 11728034 TI - Multi-sectoral approach to DOTS--the only hope. AB - This article reviews both the global and Kenyan tuberculosis (TB) situations and gives a manager's perspective of the management of a national TB control program with limited resources. It demonstrates how complex it can be to manage a control program with a 500% increase in case load and a 60% decline in real per capita health expenditure during the last 20 y. The picture is complicated even further by a high prevalence of TB among the country's nomadic population inhabiting the arid and semi-arid parts of the country. Acceptable treatment results are achieved through a combination of appropriate strategies. However, the future picture is grim unless more personnel are used to implement the DOTS strategy. PMID- 11728035 TI - Immunity to tetanus among adults in Turkey. AB - In 100 individuals > 18 y of age, living in urban or rural regions of Ankara, antitetanus immunoglobulin levels were determined using enzyme-linked immunosorbent assay and a brief questionnaire was administered. In total, 68% of subjects showed protective levels (> 0.01 IU/ml). Differences between rural (64%) and urban (72%) areas and between males (71.4%) and females (61.1%) were not significant (p > 0.05). Protective levels were found in 93.1% of subjects aged 18 30 y, 77.3% of those aged 30-40 y, 58.3% of those aged 40-50 y, 41.7% of those aged 50-60 y, 36.4% of those aged 60-70 y and 20% of those aged > 70 y. Only 35.7% of individuals aged > 50 y of age were found to be protected against tetanus (p = 0.004). The levels of antitetanus antibody in individuals vaccinated in the last 10 y were significantly higher than those in individuals vaccinated > 10 y ago and in those who did not remember the vaccination date (p = 0.009). Poor immunity against tetanus is a problem in elderly people in Turkey. PMID- 11728036 TI - The role of Clostridium difficile in childhood nosocomial diarrhea. AB - The role of Clostridium difficile was investigated in 100 children with nosocomial diarrhea. An etiologic agent was identified in 69 cases, 8 of whom had dual infection. C. difficile-associated diarrhea (Cdad) was defined in 16 children (16%). The mean age of the patients with Cdad was 5.4 y (range 2 months to 13 y) and the male:female ratio was 1.2. All cases with Cdad were on antibiotic therapy. Cdad occurred more frequently in the cases given combined antibiotic treatment than in those given single antibiotic treatment (p < 0.05). One case with neutropenic sepsis died. C. difficile was also investigated in the stool samples of 50 hospitalized children treated with antibiotics who did not develop diarrhea. C. difficile toxins A and B were found in 5 children aged < 2 y in the control group. This study shows that C. difficile is an important cause of nosocomial diarrhea in our hospital population. PMID- 11728037 TI - A 7-month outbreak of relapsing postpartum group A streptococcal infections linked to a nurse with atopic dermatitis. AB - A 7-month outbreak of 15 cases of postpartum sepsis with group A haemolytic Streptococci (GAS) was stopped when a carrier was identified. Comparing delivery dates with duty rotas revealed that the carrier had been present during delivery in 13 of the 15 cases. The epidemic GAS type, T3-13-B3264, was found in a carbuncle in her groin and in atopic dermatitis lesions behind her ears and on her eyelids. Thus, it was not the microbiological screening of staff that helped detect the carrier. The outbreak went unnoticed for 6 months, as no 2 cases were diagnosed by the same physician and 5 cases were diagnosed by different general practitioners. The main risk factors for infection were presence of the carrier relative risk (relative risk RR 47.8, 95% confidence interval (CI) 10.9-209.5) and suturing of episiotomy (RR 11.0; 95% CI 2.6-47.9). We recommend that a thorough epidemiological investigation should be carried out in every single case of GAS postpartum infection. Despite initial intravenous treatment with penicillin, 8 patients experienced > 15 recurring postpartum GAS infections, such as endometritis, wound infection, tonsillitis, erysipelas and Brodie's abscess. Eradication of GAS should be confirmed after completion of treatment. PMID- 11728038 TI - Molecular epidemiology of a multiresistant Pseudomonas aeruginosa outbreak in a paediatric intensive care unit. AB - After isolation of multiresistant (MR) Pseudomonas aeruginosa from 3 hospitalized patients in a paediatric intensive care unit (PICU), a prospective surveillance programme was established to detect infected and/or colonized patients in the hospital. Isolates were examined by means of outer membrane protein (OMP) profiles, serotyping and DNA genomic analysis using pulsed-field gel electrophoresis (PFGE). Fifty-five P. aeruginosa strains were isolated from 23 hospitalized patients during September and October 1997. The median hospital stay before isolation of P. aeruginosa was 8 d. PFGE demonstrated that the same clone infected 14 patients, 4 of whom were not hospitalized in the PICU. Susceptibility patterns and OMP profiles correlated with PFGE results in 37.8% and 36.4% of cases, respectively. Serotype O11 correlated with pattern A in 77% of cases and serotype O4 correlated with unrelated strains in 75% of cases but did not discriminate between outbreak and unrelated isolates. Extensive investigation of cultures failed to identify a reservoir of P. aeruginosa. PFGE was superior to OMP analysis and serotyping for discriminating between strains. The possible mode of acquisition for most of the patients infected with the same clone was cross contamination. PMID- 11728039 TI - Presence of Helicobacter pylori in drinking water is associated with clinical infection. AB - Helicobacter pylori was detected using molecular methods in untreated well water. The presence of H. pylori in the wells correlated with infection in consumers and with the presence of Escherichia coli, indicating fecal contamination. Consumption of untreated well water should be considered a risk factor for H. pylori infection. PMID- 11728040 TI - Evidence that Chlamydia pneumoniae affects platelet activity in patients with acute myocardial infarction and ST-segment elevations. AB - This study concerns platelet activity at myocardial infarctions and possible relationships with Chlamydia pneumoniae seroreactivity. Fourteen patients with acute myocardial infarction and ST-segment elevations were enrolled. They all received thrombolytic therapy. The subjects were examined within 24 h after hospital admission (Day 1) and after 6 months of recovery. On Day 1, C. pneumoniae IgM antibody titres were analysed and on Day 1 and during recovery C. pneumoniae IgG and soluble P-selectin were determined. P-selectin was used to estimate platelet activation. C. pneumoniae IgM titres at the infarction were closely related to both Day 1 IgG titres (r = 0.6; p < 0.05) and to IgG levels after 6 months (r = 0.8; p < 0.01). These results indicate a possible reactivation of a chronic infection. C. pneumoniae IgM was related to platelet activation. The correlation coefficient was r = 0.7 (p < 0.01) when comparing IgM titres with Day 1 plasma P-selectin. A similar relationship was found when comparing IgM and recovery P-selectin (r = 0.8; p < 0.01). The pathogen appears to contribute to platelet responses occurring during myocardial infarctions with ST-segment elevations. It is concluded that an ongoing reactivation of a chronic infection is related to increased platelet activity. PMID- 11728041 TI - Breakthrough candidaemias during empirical therapy with fluconazole in non-cancer and non-HIV adults caused by in vitro-susceptible Candida spp.: report of 33 cases. AB - The objective of this study was to assess risk factors and the outcome of breakthrough fungaemias (BFs) occurring during fluconazole (FLU) therapy in non cancer and non-HIV individuals. Thirty-three fungaemias occurring during therapy with FLU among a total of 310 fungaemias observed within a 10-y national survey were analysed. The agar disk diffusion method was used for antifungal susceptibility testing and the Vitek system for species identification. Univariate and multivariate analysis was performed to determine risk factors for BF. All BFs were due to species known to be susceptible to FLU: Candida albicans (25/33), C. parapsilosis (6/33) and C. guillermondii (2/33). The mean number of positive blood cultures per episode was 2.4. The MIC of Candida spp. to FLU was 0.5-8 mg/ml (all strains were susceptible in vitro). Neonatal age (< 4 weeks), very low birth weight, prior surgery, central venous catheter placement, artificial ventilation, total parenteral nutrition and C. parapsilosis were significantly related to BF in univariate analysis, but only central venous catheter placement was significantly related in multivariate analysis. However, the outcome of BFs and non-BFs was similar. All BFs occurred in non-HIV patients who were not previously treated with azoles, and were caused by in vitro FLU susceptible species (C. albicans and C. tropicalis). Thus factors other than in vitro susceptibility play a role in BFs. PMID- 11728042 TI - Evaluation of a dipstick test for the rapid diagnosis of imported malaria among patients presenting within the network TropNetEurop. AB - Lack of experience on the part of involved laboratory personnel frequently complicates swift diagnosis of imported falciparum malaria in non-endemic areas. Diagnostic tools based on the dipstick principle for the detection of plasmodial histidine-rich protein 2 have been marketed for several years and have been extensively evaluated. Recently, a test kit capable of detecting antigen of Plasmodium falciparum and P. vivax has been introduced. In order to evaluate this newly available tool, specimens from 664 patients were screened during the course of a prospective multicentre study within the European Network on Imported Infectious Disease Surveillance (TropNetEurop). Among the screened specimens, samples from 82 patients (12.3%) were positive for falciparum malaria using expert microscopy. A further 17 samples were positive for vivax malaria. The evaluated test kit performed with a sensitivity of 87.8% and a specificity of 99% for detection of falciparum malaria. Respective values for vivax malaria were 76.5% and 100%. Dipstick tests have the potential of improving the speed and accuracy of the diagnosis of falciparum malaria, especially if non-specialized laboratories are involved. However, decreased values of sensitivity and specificity, in comparison with expert microscopy, still impose a clear limit on the usefulness of the currently available kits. PMID- 11728043 TI - Dementia in AIDS patients in Oslo; the role of HIV encephalitis and CMV encephalitis. AB - In a well-defined population of adult AIDS patients from Oslo, we studied the correlation between clinical dementia and autopsy results. The study included 91% of all adult AIDS patients from Oslo who died between 1983 and 1996. The autopsy rate was 73% (167/229). Twenty-three percent of patients had definite dementia and 24% possible dementia. In more than half of the patients with definite dementia multinucleated giant cells were present in the brain tissue, suggesting that the dementia was due to HIV encephalitis. Diffuse damage of white matter also showed a significant association with clinical dementia. When found alone it tended to occur in symptomatic patients with a short survival time from onset of dementia until death. This indicates that diffuse damage of white matter may be an early stage of HIV encephalitis. CMV encephalitis was found in 28 cases (17%). Of these, 20 were classified as definitely or possibly demented. In 14 of these 20 cases we detected no multinucleated giant cells, suggesting that CMV caused or contributed to the dementia. Multiple logistic regression supported an association between CMV and conditions clinically classified as HIV dementia. We conclude that HIV encephalitis is the major cause of dementia in AIDS patients, but that CMV encephalitis as a cause of dementia has been underestimated. PMID- 11728044 TI - Adverse reactions to cotrimoxazole in HIV-infected patients: predictive factors and subsequent HIV disease progression. AB - The relationship between the onset of adverse events to cotrimoxazole in HIV infected patients and the subsequent development of toxoplasmosis, other AIDS defining events and survival was studied in 592 French patients who first received cotrimoxazole during the Delta trial. Low CD4+ cell count at cotrimoxazole introduction was the only factor associated with the onset of adverse reactions. The occurrence of toxoplasmosis and first AIDS-defining events were significantly and independently linked to a low CD4+ cell count at cotrimoxazole introduction (p < 0.0001) and to previous cotrimoxazole withdrawal for adverse events (p = 0.004 and p < 0.0001, respectively), but not to previous cotrimoxazole withdrawal for reasons other than adverse events, as compared to patients who did not discontinue taking cotrimoxazole during this survey. The survival rate was significantly shorter among both patients who stopped taking cotrimoxazole for adverse events and for other reasons (p = 0.03 and p = 0.0001, respectively), as compared to patients who continued to take cotrimoxazole. PMID- 11728045 TI - Changing demographics in an HIV-infected population: results from an observational cohort study in Western Denmark. AB - We present demographic data from an observational database of HIV and AIDS in the Western part of Denmark, a region with a population of 2,935,156 individuals (55.1% of the population of Denmark). Five centers in the region treat HIV positive adults; all patients attached to these centers since 1995 are included in this study. In total, 749 adult HIV-infected individuals were enrolled as of 31 December, 1999. Estimates of prevalence and incidence of HIV infection in the area were 25.9/100,000 and 2.6/100,000, respectively, which are lower than average for the country. The number of newly diagnosed HIV-infected patients remained constant during the period 1995-99, with an average of 62 diagnoses per year. The number of HIV-related deaths declined from 43 in 1995 to 15 in 1999. Of the enrolled patients, 70.9% were of Danish origin, 75% were Caucasians, 69.7% were male and 47.2% had heterosexual contact as their primary risk behavior. There seems to have been a shift in the HIV epidemic in recent years, with a higher proportion of newly diagnosed HIV patients having contracted the infection through heterosexual contact, a higher proportion being immigrants from less developed countries and newly diagnosed individuals getting older. PMID- 11728046 TI - Descending necrotizing mediastinitis caused by group A streptococcus (serotype M1T1). AB - We present a fatal case of descending necrotizing mediastinitis secondary to group A Streptococcus (serotype M1T1). Group A Streptococcus is a well-described cause of necrotizing fasciitis, but there have only been 4 previous cases documenting its involvement in descending necrotizing mediastinitis. This is the first case report to describe involvement of the M1 serotype in this condition. PMID- 11728047 TI - Propensity of Streptococcus pneumoniae for the aorta. Report of 3 cases. AB - Streptococcus pneumoniae was the unsuspected cause of a ruptured aortic aneurysm in 3 patients, as confirmed by culture of specimens obtained during surgery. A 60 y-old woman had a recently diagnosed saccular aortic aneurysm and presented with symptoms indicating a vascular catastrophe. A 66-y-old man and a 69-y-old woman were both admitted with pyrexia and abdominal pain and proper diagnosis was delayed for 4 and 15 d, respectively. All 3 patients were treated with graft insertion and antibiotic therapy for 3 months and recovered fully. PMID- 11728048 TI - Rupture of a primary aortic aneurysm infected with Shewanella alga. AB - Bacteria of the genus Shewanella are rarely implicated in bacteremia. We report a case of rupture of a primary aneurysm infected with Shewanella alga. PMID- 11728049 TI - Atrial septal defect presenting with Brucella endocarditis. AB - Brucella endocarditis, although a rare complication of brucellosis, is the main case of death attributable to this disease. An atrial septal defect (ASD) presenting with endocarditis in an adult is also extremely rare. We report the case of a male patient who presented with an infective endocarditis and was found to have an ASD. The diagnosis was proven by means of positive serology and isolation of Brucella melitensis from blood cultures and excised vegetation arising from the border of the ASD. The patient was successfully treated by means of excision of the vegetation and specific antibiotic therapy for 3 months. PMID- 11728050 TI - Sputum isolation of Wangiella dermatitidis in patients with cystic fibrosis. AB - We report a case of invasive fungal pulmonary infection in a cystic fibrosis patient. Clinical deterioration coincided with isolation of Wangiella dermatitidis from her sputum, and treatment with amphotericin B followed by voriconazole resulted in clinical improvement and sterilization of the sputum. This case suggests that W. dermatitidis may be an etiologic agent of invasive pulmonary disease in the cystic fibrosis population. PMID- 11728051 TI - Catheter-related bacteremia due to Lactobacillus rhamnosus in a single-lung transplant recipient. AB - There is still a major debate about the pathogenicity of Lactobacillus spp. and some reports emphasize that these microorganisms are never isolated from endovascular devices. In this report we present a case of catheter-related bacteremia due to L. rhamnosus in a patient who underwent a single-lung transplant. PMID- 11728052 TI - Mycoplasma pneumoniae non-pulmonary infection presenting with pharyngitis, polyarthritis and localized exanthem. AB - We report a case of pharyngitis, polyarthritis and localized exanthem in acute Mycoplasma pneumoniae infection not involving the lower respiratory tract. Diagnosis was made by means of a particle agglutination test and IgM/IgG indirect immunofluorescence assay. This case describes a clinical complex never reported before and suggests the need for a high index of suspicion in cases of atypical presentation of M. pneumoniae infection. PMID- 11728053 TI - Successful treatment of chronic disseminated candidiasis with fluconazole and a granulocyte-macrophage colony-stimulating factor combination. AB - We report a case of chronic disseminated candidiasis in a patient with acute non lymphoid leukaemia. After 12 months of combined therapy with fluconazole and granulocyte-macrophage colony-stimulating factor, lesions in the liver and spleen resolved completely. The patient has given birth to a healthy baby and has been in complete hematological remission for 3 years. PMID- 11728054 TI - Varicella-zoster virus limbic encephalitis in an immunocompromised patient. AB - A case of limbic encephalitis in a patient who had undergone prolonged immunosuppressive treatment with i.v. cyclophosphamide and oral prednisolone for a microscopic polyangeitis is reported. A brain MRI scan revealed symmetric mesial temporal lobe lesions. Studies of cerebrospinal fluid (CSF) revealed a positive PCR for varicella-zoster virus (VZV) DNA in 2 separate samples. Owing to a delay in diagnosis, intravenous acyclovir was initiated only after 11 d of symptoms. PCR of CSF for VZV DNA became negative on day 14 of treatment while brain lesions had resolved on subsequent MRI scans. Limbic encephalitis is a novel form of VZV infection. When brain imaging is suggestive of limbic encephalitis in an immunocompromised patient, PCR of CSF for VZV DNA should be performed, as early antiviral treatment may improve the outcome. PMID- 11728055 TI - Clinical ergotism induced by ritonavir. AB - Ritonavir strongly inhibits cytochrome P450, thus altering the metabolism of other drugs. We report on an HIV-positive man who, on his 13th day of ritonavir therapy, developed severe ergotism after self-administration of low doses of ergotamine. Drug interactions, the degree of responsibility of the patient and the availability of over-the-counter medications must be considered when prescribing antiretroviral therapy. PMID- 11728056 TI - Cytomegalovirus hepatitis confirmed by in situ hybridization in 3 immunocompetent infants. AB - We present 3 cases of immunocompetent infants with CMV infection who showed prolonged liver dysfunction. In all cases the CMV genome was detectable in hepatocytes using the in situ hybridization method. Combination therapy with ganciclovir (GCV) and hyperimmune gammaglobulin (HGG) was instituted in 2 cases and successfully suppressed the replication of CMV, with sustained improvement in liver function. In 1 of these cases, signals for CMV DNA were undetectable in the liver 12 months after termination of combination therapy. These results help to confirm the etiology of CMV for persistent hepatitis in immunocompetent infants using the in situ hybridization method and also show the efficacy of combination therapy with a virostatic agent, GCV, and an immune-modulating agent, HGG. PMID- 11728057 TI - Evaluation of a near-patient test and 2 enzyme-linked immunosorbent assay-based assays for detecting anti-herpes simplex virus type-2 antibodies. AB - Several type-specific serologic assays for herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), based on glycoprotein G1 (gG1) and gG2, have recently been developed. These include immunodot (POCkit HSV-2) and enzyme-linked immunosorbent assay (ELISA). The diagnostic value of POCkit HSV-2, a near-patient test, and of 2 immunoenzymatic, type-specific assays was evaluated on 122 patients attending an STD clinic. Western blot was used as the reference test. The sensitivity of POCkit HSV-2 was good but the specificity was poor, so that in a population with low seroprevalence, a positive result is likely to be a false positive. Analysis of 2 currently available HSV type-specific ELISAs yielded results suggesting that the sensitivity of these tests may also be suboptimal. PMID- 11728058 TI - Atherosclerosis and tuberculosis: are they both chronic infectious diseases? PMID- 11728059 TI - Managing time: the effects of personal goal setting on resource allocation strategy and task performance. AB - One key to understanding motivated behavior is examining the behavior of individuals as they work on multiple tasks under a time constraint. This article is an exploration of the influence of self-set goals on subsequent resource allocation to different tasks. Participants were given a variety of tasks from which they were to choose how to allocate their time and effort. Results indicated that the use of self-set goals structured the work pattern, with less switching between tasks relative to the work pattern of a group of participants who did not set goals. In addition, those who set goals reported less task related cognitive interference, indicating that they were not as distracted while they worked. Participants who did not set goals, however, performed at a higher level on some of the tasks. It is suggested that self-set goals may often be chosen at an easily attainable level, creating a structured and focused work environment but not necessarily eliciting the motivational properties typically associated with goal setting. PMID- 11728060 TI - Hedonic intensity of disappointment and elation. AB - This research is an investigation of the hedonic intensities of elation and disappointment following the outcomes of risky gambles using two principles: disappointment aversion and the principle of limited emotion processing. Disappointment aversion implies a stronger impact of disappointment compared with elation; the principle of limited emotion processing predicts a smaller impact of elation if it occurs with a real gain, and a smaller impact of disappointment if it occurs with a real loss. Both principles support each other in the gain domain but operate against each other in the loss domain. It was predicted that disappointment would outweigh elation in the gain domain. For the loss domain, this question was left open to empirical scrutiny. Participants were provided with hypothetical gambles and were required to imagine having won, not won, lost, or not lost, money. Results supported the prediction for the gain domain; mixed results emerged for the loss domain. The model adds to the understanding of the cognitive and emotional processes following the outcomes of risky events. PMID- 11728061 TI - Dimensions and aspects of longing: age and gender differences in Swedish 9-, 12-, and 15-year-old children. AB - Longing can be defined as a secondary emotion, as a blend of the primary emotions of love and sadness. There are several possible dimensions and aspects of longing (O. Holm, 1999). Both age and gender differences are well documented in earlier research on other emotions. In the present investigation, 122 girls and 120 boys, ages 9, 12, and 15 years, in compulsory school in Sweden, answered a questionnaire about dimensions and aspects of their own longing. The results showed both age and gender differences. Girls, especially in the 15-year-old group, experienced longing significantly more than boys. The results were interpreted as generally in accordance with what is known from earlier research on other emotions. PMID- 11728062 TI - Situational leadership style as a predictor of success and productivity among Taiwanese business organizations. AB - The present study was an evaluation of the impact of Taiwanese leadership styles on the productivity of Taiwanese business organizations. Specifically, it looked at the impact that both adaptive and nonadaptive leaders have on 6 measures of productivity: absenteeism, turnover rate, quality of work, reject rates, profitability, and units produced. The results indicated that the greater the level of adaptability, the more productive the organization is likely to be. Although not all of the computed correlations were statistically significant, they were all in the predicted directions. In particular, the findings for units produced and reject rates were consistently statistically significant. The study was also an examination of the usefulness of the Leadership Effectiveness and Adaptability Description (LEAD) questionnaire (P. Hersey & K. Blanchard, 1988), which appeared to be an accurate predictor of adaptability and valid for use in Taiwan. The final part of this study was an investigation of whether successful companies were more likely to have a greater percentage of adaptive leaders than unsuccessful companies. The data supported this expectation, although it is suggested that caution be used in the interpretation of this particular finding because it could have several different explanations. Overall, the evidence supported the value of adaptive leadership styles in high-technology industries in Taiwan. PMID- 11728063 TI - Body attitude, gender, and self-concept: a 30-year perspective. AB - Two comparable samples of college men and women from 1966 and 1996 were compared using the Kurtz Body Attitude Scale (BAS; R. M. Kurtz, 1966). As predicted, women in 1996 reported a significantly more negative body attitude than women in 1966 did. No significant differences in the 2 samples of men were found. With the 1996 sample, the relationship between body attitude and self-esteem was also examined using Marsh's Self-Description Questionnaire-III (SDQ-III; H. W. Marsh & R. O'Neill, 1984). There was a significant relationship between body attitude and general self-esteem, and there were also complex gender differences in the relationships of various SDQ-III facets to body attitudes. PMID- 11728064 TI - Factors predicting future time orientation for romantic relationships with the opposite sex. AB - The author examines the possible connection of predictors of future time orientation to romantic relationships with the opposite sex using the Future Time Orientation in Romantic Relationships Scale (FTORR; B. Oner, 2000b). The variables suggested as correlates with FTORR scores in B. Oner's (2000a) study were subjected to regression analysis. The results indicated that relationship satisfaction and an eagerness to break up negative relationships were negatively related to FTORR scores, whereas being a woman, experiencing jealousy, and the degree of selectivity in choosing a dating partner were positively related to FTORR scores. Caution and relationship commitment both predicted FTORR scores through the mediating effect of relationship satisfaction. The author discusses the implications of the findings. PMID- 11728065 TI - The relationship between life satisfaction, self-consciousness, and the Myers Briggs type inventory dimensions. AB - The study was an investigation of the relationship between psychological well being, life satisfaction, self-consciousness, and the four Myers-Briggs Type Indicator dimensions (MBTI; I. B. Myers & M. H. McCaulley, 1985). The participants were 97 college students (79 women and 18 men whose mean age was 31.4 years). All the students were administered four instruments, the Psychological Well-Being Inventory (C. D. Ryff, 1989), the Satisfaction With Life Scale (E. Diener, R. A. Emmons, R. J. Larsen, & S. Griffin, 1985), the Self Consciousness Scale-Revised (M. F. Scheier & C. S. Carver, 1985), and the MBTI (Form G Self-Scoring). MANOVAs revealed significant differences on three of the four dimensions of the MBTI with extraverts showing higher psychological well being and life satisfaction and lower self-consciousness than introverts. Intuition types scored higher in psychological well-being and lower in self consciousness than Sensing types. Judging types scored higher in psychological well-being than Perceiving types. Correlational analyses showed that most dimensions of psychological well-being were negatively related to self consciousness. The relationship between life satisfaction and personality variables is discussed. PMID- 11728066 TI - Is self-reflectiveness an unhealthy aspect of private self-consciousness? AB - A number of researchers (M. Conway & C. Giannopoulos, 1993; P. J. Watson & M. D. Biderman, 1993) have suggested that self-reflectiveness is an unhealthy aspect of private self-consciousness related to psychopathology. However, these studies did not control for the significant correlation that exists between self reflectiveness and subscale factors of public self-consciousness: style consciousness and appearance consciousness. The purpose of this research was to address these interrelationships by comparing correlational results with those obtained from multiple regression analyses. Participants (N = 111) completed the Self-Consciousness Scale (A. Fenigstein, M. F. Scheier, & A. H. Buss, 1975) and the NEO-Five Factor Inventory (P. T. Costa & R. R. McCrae, 1992). The results suggested that when the effects of style consciousness and appearance consciousness are controlled, the relationship between self-reflectiveness and neuroticism is nonsignificant. Furthermore, multiple regression analyses suggested a significant relationship between self-reflectiveness and low levels of agreeableness. Aspects of self-consciousness may be better understood in the context of overlapping domains that consider both common and unique variance. PMID- 11728067 TI - The Irrational Beliefs Survey, one factor or four? A replication of Mahoney's (1997) findings. PMID- 11728068 TI - The Finnish Arthroplasty Register: report of the hip register. AB - The Finnish Arthroplasty Register was established in 1980. Between 1980 and 1999, 62,841 primary and 12,224 revision total hip arthroplasties (THA) were recorded. The annual number of both primary and revision THA has increased: in 1999, the incidence of primary THAs was 93/100,000. 174 implant designs have been used, but the 6 commonest implants comprised 82% in 1999. Since the late 1980s, more than 40% of the hips were inserted without cement. Over 47% of the cementless primary hip prostheses were used in patients younger than 60 years and over 93% of the cemented primary hips were used in patients 60 years or older. The 10-year survival rate was 72 (95% CI 67-76)% in patients younger than 55 years and 90 (89 91)% in patients older than 70 years. The commonest reasons for revision were aseptic loosening (65%), dislocation (9%) and infection (7%). In revisions, the 5 year survival of the cementless hip prosthesis improved over time: it was 85 (82 87)% in 1985-1989, 89 (88-91)% in 1990-1994 and 92 (88-95)% in 1995-1999. There are striking differences between the Arthroplasty Registers of Scandinavia as regards the end-point definition of survival. The Finnish Arthroplasty Register considers all reasons for revisions as the end-point of survival, but the Swedish register takes into account only aseptic loosening, so direct comparisons between registers are not possible. Recent data from the Finnish Arthroplasty Register indicate that the results of total hip replacements are improving in Finland. With the civic registration number, one can link and match data files. For example, with use of the Finnish Cancer Register, we found no increase in the risk of cancer after a THA. PMID- 11728069 TI - Blood conservation with tranexamic acid in total hip arthroplasty: a randomized, double-blind study in 40 primary operations. AB - We performed a randomized, double-blind study on the effect of tranexamic acid on blood loss and blood transfusions in 40 primary total hip arthroplasties. Tranexamic acid, 10 mg/kg body weight, or placebo, was given intravenously just before the operation. Blood loss during the operation and postoperatively into the drains was recorded, as also were blood hemoglobin concentrations. Ultrasound examination 1 week postoperatively was done to estimate the blood loss due to remaining hematomas. Total (operation + drain) blood loss was 0.76 (95 CI 0.63 0.89) L in the tranexamic acid group as compared to 1.0 (CI 0.81-1.2) L in the placebo group (p = 0.03). The number of blood transfusions during the day of operation was 2 vs. 10 (p = 0.07) and the total number during the hospital stay was 5 vs. 13 (p = 0.2). 1 patient in each group had a pulmonary embolism. PMID- 11728070 TI - Poor outcome of 44 cemented total hip arthroplasties with alumina ceramic heads: clinical evaluation and retrieval analysis after 10-16 years. AB - We evaluated the long-term results of cemented total hip arthroplasty with alumina ceramic heads in 39 patients (44 hips) from 1981 to 1985. The study comprised 8 men (8 hips) and 31 women (36 hips). Their mean age was 54 (37-76) years and mean weight 52 (34-93) kg. The preoperative diagnoses were osteoarthrosis in 34 patients (38 hips), rheumatoid arthritis in 3 (4 hips) and idiopathic osteonecrosis of the femoral head in 2 (2 hips). The mean duration of follow-up was 13 (10-16) years. At this time, 6 cases had been revised because of aseptic loosening (5 hips showed loosening of both components and 1 acetabular loosening alone). Kaplan-Meier survivorship analysis with radiographic loosening as end-point showed 10-year survival rates of 75% for the acetabular component and 86% for the femoral. The mean linear wear rate of polyethylene was 0.10 mm/year radiographically--i.e., about the same as reported for many metal to polyethylene bearings. 5 alumina ceramic heads were retrieved and showed excellent surface roughness and roundness without scratches. However, debris stuck to the gaps was detected. The debris may act as third-bodies and affect polyethylene wear. PMID- 11728071 TI - Charnley cups retained during revision of the femoral component: repeated clinical and radiographic observations of 49 cups for 7-15 years. AB - This article concerns 49 consecutive polyethylene cups retained in cemented first revisions of the femoral component and evaluated on several occasions 7-15 years after the index operation. At the last follow-up 5 cups were revised, 3 were radiographically loose, 1 was revised because of recurrent dislocation and 1 stable cup was revised because the surgeon wished to use a 28 mm head. Moreover, 1 unrevised cup was radiographically loose. This corresponded to a mechanical failure rate of 0.1 (5/49) and revision rate of 0.1 (4/49). 3 cups which were radiographically loose (100% radiolucency) had not been revised when the stem was exchanged. If these cups were excluded, the revision rate would have dropped to 0.02 (1/46) and the mechanical failure rate to 0.04 (2/46). This indicates the importance of careful preoperative radiographic evaluation on both AP and lateral views. Progression of radiolucent lines always occurred as an increase in extension from the periphery to the central region of the interface. In conclusion, Charnley cups with non-circumferential radiolucent lines and little, if any, wear can be retained without compromising the longevity of the implant. However, it should be stressed that this conclusion is based on a patient population in which four-fifths of the patients at the index operation were 65 years of age or older. PMID- 11728072 TI - Prevention of heterotopic ossification after total hip replacement: a prospective comparison of indomethacin and salmon calcitonin in 60 patients. AB - We did a prospective consecutive study of prophylaxis for heterotopic ossification (HO) comparing indomethacin (100 mg/day) and salmon calcitonin (3 MRC-U/kg/day) for 14 days. Each group consisted of 30 patients. 19 patients in the indomethacin group and 2 in the calcitonin group developed HO. We conclude that use of calcitonin in the prophylaxis of HO after total hip replacement is more effective than indomethacin. PMID- 11728073 TI - Stiffness and compactness of morselized grafts during impaction: an in vitro study with human femoral heads. AB - The mechanical properties of the impacted material is important when using impaction bone grafting in revision arthroplasty. We did an in vitro study to monitor the stiffness and compactness of morsellized bone grafts during impaction. Using human osteoarthrotic femoral heads as the bone source we prepared 3 types of morselized grafts with the same bone mill: (1) purely cancellous grafts, (2) cortico-cancellous grafts and (3) cortico-cancellous bone with cartilage remnants. 5 g of bone samples were progressively impacted up to 150 times in a contained cavity. All types of grafts became stiffer and stiffer during the first 30 impactions. After 30 impactions, the grafts' compactness continued to increase slowly without concomitant changes in their stiffness. Over impaction was not useful, but did not jeopardize the implant stability. The presence of cartilage remnants considerably compromised the increase in stiffness. In contrast, the addition of cortical bone from the femoral neck did not affect the impaction and resultant stiffness. PMID- 11728074 TI - Knee pain caused by a fiducial marker in the medial femoral condyle: a clinical and anatomic study of 20 cases. AB - After 2-pin-based ROBODOC hip arthroplasty procedures, 10 of 18 patients reported persistent severe pain at the site of pin implantation in the medial femoral condyle. In a cadaver study, we found that the infrapatellar branch of the saphenous nerve, the saphenous nerve and the anterior cutaneous branches of the femoral nerve had been injured by the pins. At least one of these nerves was injured in 11 of the 20 specimens examined. Our findings indicate that the knee pain may be partly caused by injuries to these nerves. PMID- 11728075 TI - No bone ingrowth into the tibia tunnel in anterior cruciate ligament reconstructed patients: a 1-year prospective quantified CT study of 10 patients reconstructed with an autologous bone-patellar tendon-bone graft. AB - 10 patients with major instability symptoms due to an acute anterior cruciate ligament injury were operated on with a bone-patellar tendon-bone reconstruction. Tibial condyle bone mineral density (BMD), bone ingrowth and changes in diameter in the tibia bone tunnel were studied with quantified computed tomography (QCT) postoperatively and after 1, 3, 6 and 12 months. We found no sign of bone ingrowth in the form of increased bone mineral density (BMD) in the bone tunnels in any of the patients. The tunnel diameter increased in all patients during the first postoperative months. After 1 year, 5 patients had a smaller diameter than at the first postoperative examination, 2 had the same diameter as immediately after surgery and 2 patients had a larger diameter. A sclerotic zone developed in all patients along the perimeter of the tunnel during the 3-6 months of follow up. The BMD in the tibial condyle decreased at 3 months; it then increased, but between 6 and 12 months, it levelled out and was slightly lower than postoperatively. In conclusion, we found no growth of bone into the tunnel and tendinous part of the graft during the first postoperative year. PMID- 11728076 TI - The Richards type II patellofemoral arthroplasty: 26 cases followed for 1-20 years. AB - We followed 26 Richards type II patellofemoral arthroplasties in 24 patients (19 women) for a mean of 11 (1-20) years. Their mean age was 59 (22-90) years. The preoperative diagnoses were primary patellofemoral arthrosis in 17 cases and secondary arthrosis in 9 (8 malalignment, 1 patellar fracture). Patellectomy was later performed for persistent pain or patellar malalignment in 3 cases and a conversion to a total knee arthroplasty for progressive tibio-femoral degeneration or patella malalignment in 2. The mean Knee Society knee score for 21 knees at follow-up was 90 (65-100) points. The patients rated the results of surgery in 9 knees as excellent, 7 good, 4 improved, and 1 unimproved at follow up. None of the implants showed signs of loosening or infection. The Richards type II patellofemoral arthroplasty yields acceptable long-term results in patients with isolated end-stage patellofemoral osteoarthrosis. Patient selection and patella alignment are important. PMID- 11728077 TI - Correlation of synovial fluid cytokine levels with histological and clinical parameters of primary and revision total hip and total knee replacements. AB - We retrieved synovial tissue and fluid samples from patients undergoing primary total hip replacement (THR) (n 15), revision of aseptically loose THR (n 12), primary total knee replacement (TKR) (n 13) and revision of aseptically loose TKR (n 6). Several histological parameters were assessed on a relative scale of 14. Primary TJRs were clinically evaluated for degree of osteoarthrosis. Revision TJRs were assessed for migration of the implant, gross loosening and the degree of radiolucency. Cytokine levels in synovial fluid were determined with ELISA. All cytokines were significantly higher in revision TJRs than in primary replacements, as were the degree of macrophage and giant cell infiltration. We found no relationship between any clinical variable and the levels of any cytokine, but migration of the implant was related to the presence of PE debris. A significant correlation was seen between the presence of macrophages and the levels of IL-1beta, IL-8 and IL-10, but not IL-6. No differences were noted between hips and knees for any of the variables, except in the levels of IL-6, where higher levels were found in THRs. These results suggest a unique role for IL-6 that requires further investigation. PMID- 11728078 TI - The effect of flurbiprofen and tiaprofenic acid on serum cytokine levels of patients with osteoarthrosis. AB - 39 patients with active knee osteoarthrosis, chosen according to ACR criteria, were assigned to receive flurbiprofen (n 12, 2 x 100 mg), tiaprofenic acid (n 14, 2 x 300 mg) and placebo (n 13) in a 3-week, placebo-controlled study. All patients completed the study, and both medications were found to be effective: improvement occurred in the clinical signs. These drugs reduced the TNF-alpha levels. Flurbiprofen especially affected the IL-6 levels. Our findings indicate that NSAIDs may be effective in the etiopathogenesis of osteoarthrosis. PMID- 11728079 TI - The Swedish Knee Arthroplasty Register 1975-1997: an update with special emphasis on 41,223 knees operated on in 1988-1997. AB - From 1975, when the Swedish Knee Arthroplasty Register (SKAR) started, until the end of 1997, 57,533 primary arthroplasties and revisions have been registered. Recently, the register underwent a comprehensive validation and update regarding revisions. We now report on general demographic and epidemiological data for the whole period and on the survivorship of arthroplasties performed in Sweden during 1988-1997. During this 10-year period, 41,223 primary knee arthroplasties were performed on 34,877 patients. We found, as in our earlier reports, that survivorship was affected by patient-, time-, implant- and method-related factors but, apart from an overall higher cumulative revision rate, general conclusions reported from the register in recent years appeared to be unaffected. PMID- 11728080 TI - Tuberculosis of the sternoclavicular joints. AB - From May 1991 to December 1997, we treated 9 patients with tubercular arthritis in 10 sternoclavicular joints. The patients presented with a painful swelling (7 joints), painless swelling (2 joints) and a painless (?) discharging sinus (1 joint) having a mean duration of symptoms of 13 (6-32) months. The diagnosis was made with fine-needle aspiration or open biopsy. In 1 patient debridement of the joint was combined with open biopsy. All patients were initially put on a 4-drug regimen of antitubercular therapy (ATT). 2 joints not responding to closed treatment were surgically debrided after 2-3 months of ATT. Total duration of ATT was 14-18 months. At final follow-up after average 4.5 (1.5-7.5) years, all lesions had healed. 3 patients had mild limitation of shoulder motion, with no pain, and 2 patients had a cosmetically ugly scar at the site of the sinus or biopsy. PMID- 11728081 TI - Early active rehabilitation after surgery for lumbar disc herniation: a prospective, randomized study of psychometric assessment in 50 patients. AB - In a randomized study, using psychometric assessment, we evaluated two training programs before and after surgical treatment of lumbar disc herniation. 26 patients were treated according to an early active training program (treatment group). 24 patients followed a traditional less active training program (control group). Before surgery, the patients filled in the following questionnaires 3 and 12 months after surgery: Multidimensional Pain Inventory (MPI), State and Trait Anxiety Inventory, and Beck Depression Inventory. Pain was assessed by the patient's pain drawing and a visual analog scale. Both groups improved as regards pain severity and state of anxiety. The MPI parameter, pain interference, improved more in the early active treatment group than in the control group. This suggests that the early active training program has a positive effect on the way patients cope with pain in their daily lives. PMID- 11728082 TI - Comparison of osteopenia after gastrectomy, ovariectomy and prednisolone treatment in the young female rat. AB - Rat models of osteopenia include ovariectomy and long-term glucocorticoid treatment. Although ovariectomy produces significant trabecular bone loss after 2 weeks, long-term glucocorticoid treatment has been reported to cause osteopenia in some studies but not in others. In the present 8-week-study, we compared the osteopenia associated with gastrectomy (GX) to that induced by ovariectomy (OVX) or prednisolone (PRE) treatment. Female Sprague-Dawley rats (10 weeks old) were subjected to GX, OVX, PRE treatment or SHAM operation. At the end of the study, calvariae, femurs and fifth lumbar vertebrae (L5) were collected and subjected to bone density measurement (femur and L5), transillumination (calvaria) and histomorphometry (calvaria and femur). Bone density was reduced in L5 and the distal femur in the OVX and GX groups, but not in the PRE group. Transillumination of the calvaria showed marked bone loss in the GX rats, but not in the other groups. Morphometric analysis of the femur revealed reduced trabecular bone volume, trabecular thickness, trabecular number and osteoclast number, but increased osteoclast surface (expressed as per cent of the trabecular bone surface covered by osteoclasts) in the GX and OVX rats. The PRE rats seemed unaffected. Cortical thickness was reduced in the GX rats, but not in the other groups. The findings indicate that GX induces osteopenia in, e.g., femur and vertebra of a magnitude similar to or greater than that induced by OVX, while at the same time inducing osteopenia in the calvaria. Although osteoclast activation seems to contribute, the precise mechanism underlying the GX-evoked osteopenia remains obscure. PMID- 11728083 TI - Stump lengthening after hip disarticulation using a modular endoprosthesis in 5 patients. AB - To reduce the disability after hip disarticulation, we developed a special surgical procedure in patients having a proximal femoral tumor with a large tumor involving the sciatic nerve or neoplasms involving the tibia and femur. The hip was disarticulated, but we preserved a musculocutaneous flap. A modular endoprosthesis was then placed in the acetabulum or, in case of an extraarticular resection of the hip joint, it was placed in the iliac bone. A trevira tube was used for reconstruction of the joint capsule and fixation of soft tissues. We performed this procedure in 5 patients who had a good functional outcome. PMID- 11728084 TI - Obturator neuropathy due to intrapelvic extrusion of cement during total hip replacement--report of 2 patients. PMID- 11728085 TI - Bo Nilsson--1933-2001. PMID- 11728086 TI - An introduction to "Water in the cell": tamed Hydra? PMID- 11728087 TI - Life's matrix: water in the cell. AB - Water is a profoundly unusual liquid, and its peculiarities may make it uniquely suited to act as life's matrix. Even if this were not so, however, we should expect the effects of nanometre-scale confinement and inhomogeneities owing to surface effects to alter the liquid's properties in the cell relative to those in the bulk. Whether water's unusually high degree of local structure makes such influences even more marked than for 'normal' liquids remains an open question, with potentially important consequences for biomolecular interactions. PMID- 11728088 TI - Water, treated as the continuous liquid in and around cells. AB - In the quantitative treatment of non-covalent inter- and intra-cellular interactions taking place in water, in vitro as well as in vivo, it is essential to treat the surrounding and pervading liquid medium as the continuous medium. In the close vicinity of inter- and intra-cellular surfaces and of biopolymers the various different non-covalent forces may locally alter the structure of water in a number of ways, but these local structural changes can be quantitatively taken into account. The operative forces are: Lifshitz-van der Waals (LW) forces. Lewis acid-base (AB) forces and electrostatic (EL) forces. Of these, the AB forces are generally the preponderant ones, in aqueous media. This is due, inter alia, to the strong cohesive and adhesive hydrogen-bonding interactions typically occurring in and by water. Among the strong AB interactions occurring in water are hydrophobic attraction (the hydrophobic effect) and hydrophilic repulsion (hydration pressure). Also treated is the function of LW, AB and EL forces in: hydration; in the stability of particle and cell suspensions, the solubility of biopolymers, small organic solutes, and electrolytes; and in specific ligand receptor (such as antigen-antibody) interactions. PMID- 11728089 TI - High and low density intracellular water. AB - The proposal that liquid water consists of microdomains of rapidly-exchanging polymorphs of high and low density is examined for its impact upon roles of water in biology. It is assumed that the two polymorphs persist in solution and adjacent to surfaces and that solutes partition asymmetrically between them. It transpires that chaotropes are solutes which partition preferentially into low density water and displace the water equilibrium toward the high density polymorph. Kosmotropes. both ionic and non-polar, partition into high density water and induce low density water. Displacement of the water equilibrium at constant temperature and pressure has a thermodynamic cost which can be high. This appears to be a dominant factor in folding of proteins and DNA, aggregation of biopolymers and insolubility of non-polar kosmotropes. Cells control both the concentration of proteins and the selection of small solutes to produce an intracellular environment most conducive to co-ordinated enzyme function. Intracellular water has similar microdomains to bulk water, but surfaces and solutes redistribute them. Average properties, as measured by NMR are similar, but local properties on a nm scale may differ widely. Enzymes apparently use these local differences to activate cations for transport, induce movement and for synthesis. PMID- 11728090 TI - Physical aspects of the weakly hydrated protein surface. AB - We review the physical properties of water on the surface of weakly hydrated proteins and present some theoretical models used to understand them. The first part concerns mainly structural properties and introduces a model for two dimensional clusters of water molecules. The second part is devoted to dynamical properties of the hydrated protein surface. Dielectric measurements which provide an evidence for proton conductivity due to the percolation of the network of surface water molecules and for the glass dynamics of migrating protons when temperature is lowered are reviewed. These results can be associated with the concept of frustration and analyzed with two models, an Ising model to describe the proton jumps and the model of two-dimensional surface water which exhibits a glassy dynamiques of the water molecules. Biological implications of these properties of hydration water are briefly discussed. PMID- 11728091 TI - Protein denaturation described by a two-state structural model of liquid water. AB - Our laboratory has been involved in expounding an outer neighbor molecular-level structural theory for liquid water in the supercooled and ordinary thermodynamic regimes. This theory, which depends on transformations with increasing temperature or pressure, is consistent with all the properties of this substance, including the ten or so "anomalies", and has been recently used to explain the effect that surrounding water has on proteins. For example, the sharp decrease in the hydration entropies of polar groups can be explained through a consideration of the promotion of ice-Ih-type bonding structure at the expense of the less stable ice-II-type bonding structure. These structural transformations occur in the local neighborhood of the polar group. In this paper we discuss this outer neighbor two-state structural theory for liquid water, the role it plays in explaining water's anomalous properties and its description of protein denaturation both as a function of temperature and pressure. PMID- 11728092 TI - Large-scale networks of hydration water molecules around proteins investigated by cryogenic X-ray crystallography. AB - The structures at protein-water interface, i.e. the hydration structure of proteins, have been investigated by cryogenic X-ray crystal structure analyses. Hydration structures appeared far clearer at cryogenic temperature than at ambient temperature, presumably because the motions of hydration water molecules were quenched by cooling. Based on the structural models obtained, the hydration structures were systematically analyzed with respect to the amount of water molecules, the interaction modes between water molecules and proteins, the local and the global distribution of them on the surface of proteins. The standard tetrahedral interaction geometry of water in bulk retained at the interface and enabled the three-dimensional chain connection of hydrogen bonds between hydration water molecules and polar protein atoms. Large-scale networks of hydrogen bonds covering the entire surface of proteins were quite flexible to accommodate to the large-scale conformational changes of proteins and seemed to have great influences on the dynamics and function of proteins. The present observation may provide a new concept for discussing the dynamics of proteins in aqueous solution. PMID- 11728093 TI - Water and cellular folding processes. AB - Proteins require a unique, native structure to perform their functions. Water molecules play an important role to develop and maintain this three-dimensional structure. Water is also necessary for several forms of enzyme catalysis, and is a constituent of many protein-protein, protein-DNA, or protein-RNA interfaces. Larger proteins acquire their native structure in a complicated folding pathway having several folding traps. Recent data indicated a key role of water molecules in this process. Protein flexibility, structural rearrangements, conformational transitions all require the fluctuating changes in hydrogen bond structure provided by interacting water molecules. Besides proteins, RNA and DNA structure is also heavily influenced by the presence of water. This review summarizes the important aspects of these fields, and draws attention to several open questions and hypotheses. PMID- 11728094 TI - Hydration effects on DNA double helix stability modulates ligand binding to natural DNA in response to changes in water activity. AB - In this work we present evidence that water molecules are actively involved on the control of binding affinity and binding site discrimination of a drug to natural DNA. In a previous study, the effect of water activity (a(w)) on the energetic parameters of actinomycin-D intercalation to natural DNA was determined using the osmotic stress method (39). This earlier study has shown evidence that water molecules act as an allosteric regulator of ligand binding to DNA via the effect of water activity on the long-range stability of the DNA secondary structure. In this work we have carried out DNA circularization experiments using the plasmid pUC18 in the absence of drugs and in the presence of different neutral solutes to evaluate the contribution of water activity to the energetics of DNA helix unwinding. The contribution of water to these independent reactions were made explicit by the description of how the changes in the free energy of ligand binding to DNA and in the free energy associated with DNA helix torsional deformation are linked to a(w) via changes in structural hydration. Taken together, the results of these studies reveal an extensive linkage between ligand binding affinity and site binding discrimination, and long range helix conformational changes and DNA hydration. This is strong evidence that water molecules work as a classical allosteric regulator of ligand binding to the DNA via its contribution to the stability of the double helix secondary structure, suggesting a possible mechanism by which the biochemical machinery of DNA processing takes advantage of the low activity of water into the cellular milieu. PMID- 11728095 TI - Hydration and structural alterations of A-DNA. Implications for the accuracy of DNA replication. AB - The accuracy of replication, transcription and translation depends on the correct pairing of complementary bases of nucleic acids. However, sequence dependent structural variations of double helices may interfere with the fidelity of these processes in altering base pair conformations. Irregular conformations of the template-primer DNA duplex can induce replication errors in altering the structure of the polymerase active site. However, in most DNA polymerases, water exclusion induces the duplex to adopt the A-conformation. In attenuating the structural variability, the A-form enhances replication fidelity. How hydration and DNA sequence modulate the structure of A-DNA is therefore a relevant question for better understanding the mechanisms of sequence directed mutagenesis. This study is focused on the relationships between hydration and the distortion of A DNA double helices. The crystallographic analysis of the hydration pattern of A DNA decamers reveals that the subtle interplay of DNA sequence and hydration helps to stabilize unusual structures. These findings are discussed in the context of the accuracy of replication. PMID- 11728096 TI - Role of hydration of polyhydroxy compounds in biological systems. AB - The spin-lattice relaxation times, T1, of H2(17)O have been measured for aqueous solutions of 9 polyols as a function of the concentration at 25 degrees C. The dynamic hydration number, nDHN, for polyols were obtained. The values of nDHN increased with an increase in the number of OH groups and depended on the conformation of isomers. The value of nDHN for inositol was the largest. This means that the thermal motion of water molecules around inositol is most inhibited among the polyols studied. The increment of thermal stability of proteins by polyols and sugars, the equilibrium distribution coefficients of polyols and sugars in the denatured hemoglobin solution, and the effect of sugars on the osmotic flow of water were expressed by linear relation of nDHN. On the bases of these relations, a systematic treatment for the effect of hydration of polyols and sugars on these phenomena is discussed. PMID- 11728097 TI - Osmotic mass transfer in the yeast Saccharomyces cerevisiae. AB - This paper reviews the passive mechanisms involved in the response of a yeast to changes in medium concentration and osmotic pressure. The results presented here were collected in our laboratory during the last decade and are experimentally based on the measurement of cell volume variations in response to changes in the medium composition. In the presence of isoosmotic concentration gradients of solutes between intracellular and extracellular media, mass transfers were found to be governed by the diffusion rate of the solutes through the cell membrane and were achieved within a few seconds. In the presence of osmotic gradients, mass transfers mainly consisting in a water flow were found to be rate limited by the mixing systems used to generate a change in the medium osmotic pressure. The use of ultra-rapid mixing systems allowed us to show that yeast cells respond to osmotic upshifts within a few milliseconds and to determine a very high hydraulic permeability for yeast membrane (Lp>6.10(-11) m x sec)-1) x Pa(-1)). This value suggested that yeast membrane may contain facilitators for water transfers between intra and extracellular media, i.e. aquaporins. Cell volume variation in response to osmotic gradients was only observed for osmotic gradients that exceeded the cell turgor pressure and the maximum cell volume decrease, observed during an hyperosmotic stress, corresponded to 60% of the initial yeast volume. These results showed that yeast membrane is highly permeable to water and that an important fraction of the intracellular content was rapidly transferred between intracellular and extracellular media in order to restore water balance after hyperosmotic stresses. Mechanisms implied in cell death resulting from these stresses are then discussed. PMID- 11728098 TI - Osmolytes responsible for volume reduction under isosmotic or hypoosmotic conditions in Barnacle muscle cells. AB - In numerous animal cells, experimental manipulations that increase the intracellular free Ca2+ concentration induce cell volume reduction. This may occur under isosmotic conditions, e.g. when external Ca2+ (Ca(o)) is replaced by Mg2+ (42) or during exposure to hypoosmotic conditions (i.e. regulatory volume decrease, RVD) in the presence of Ca(o). We determined the osmolytes responsible for volume reduction under isosmotic and hypoosmotic conditions in barnacle muscle cells. Organic osmolytes (i.e. free amino acids and methylamines) and inorganic ions accounted for approximately 78% and 22% of the intracellular isosmotic activity, respectively. Isosmotic Ca(o) removal induced a net loss of KCI (with a ratio of 1K:1Cl) and free amino acids (FAA, mainly glycine and taurine). During RVD. the same ions (but in a proportion of 2K:1Cl) and FAA were lost. Since RVD was accompanied by extracellular alkalinization, the 2K:1Cl loss may be explained by the presence of a K+/H+ exchanger (or K+-OH- co-transporter) or Cl-/OH- exchanger. The lack of RVD in the absence of Ca(o) cannot be attributed to the loss of intracellular osmolytes during isosmotic Ca(o) removal because addition of Ca(o) during cell swelling promoted RVD. PMID- 11728099 TI - Osmotic properties of internally perfused barnacle muscle cells. I. Isosmotic conditions. AB - Barnacle muscle cells regulate their volume when exposed to anisotonic conditions. Due to their large size, these cells can be internally perfused. Interestingly, perfused cells maintain their volume regulatory properties (17,21). Thus, the osmotic properties of barnacle muscle cells can be studied under conditions in which the intracellular and extracellular osmolalities, the membrane potential (V(M)), the cell volume and the intracellular pressure can all be measured simultaneously. In this manuscript we report the effect that various rates of isosmotic (1000 mOsm x kg H2O(-1)) intracellular perfusion have on cell volume, intracellular pressure, intracellular osmolality, V(M), and the apparent sarcolemmal hydraulic water permeability (L'p). Replacement of the cytosol with the perfusate at a perfusion rate of 0.83 microl x min(-1) took 120 min. During this transition period, the cell volume increased from 45.1+/-6.9 microl to 73.7+/-5.8 microl, the intracellular osmolality decreased from 1406+/-133 to 1188+/-64 mOsm x kg H2O(-1), and the intracellular pressure underwent a transient drop of 2.8 cm H2O. After 2.5 hr of continuous perfusion at 0.83 microl min(-1), the above mentioned parameters reached steady values: the L'p was 1.35 x 10(-5) cm x sec(-1) x Osm(-1) x kg H2O(-1); cell volume was 67.2+/-6 microl; the intracellular osmolality was 1052+/-10 mOsm x kg H2O(-1); the intracellular pressure was 5.6+/-0.4 cm H2O; V(M) depolarized slowly at a rate of 0.03 mV x min(-1). Stepwise increases in the rate of perfusion (from 0.83 to 3.18 microl min(-1)) produced reversible increases in the intracellular pressure, L'p and cell volume and decreases in intracellular osmolality. We conclude that intracellular perfusion: i/ produces a transient removal of intracellular osmotically active components; ii/ promotes sarcolemmal water filtration; iii/ induces a laminar flow of perfusate at the center of the cell, and iv/ enables calculations of sarcolemmal L'p values under isosmotic conditions. PMID- 11728100 TI - Temperature effects on cell-functioning--a critical role for vicinal water. AB - Interfacially modified water ('vicinal water') appears to extend over distances of several molecular diameters from an adjacent interface and possesses highly unusual thermal properties. Specifically, the vicinal water undergoes relatively abrupt changes over narrow temperature intervals centered around 15 degrees, 30 degrees, 45 degrees and 60 degrees C and these changes most likely reflect higher order (structural) phase transitions. Vicinal water occurs at all interfaces (independent of specific surface characteristics) and will therefore also be present in all biological cells. As a result, the effects of vicinal water ramify through all of cell biology and this explains many unexpected thermal anomalies seen in the temperature responses of a great variety of living systems. Thus, the concept of the thermal anomalies may explain such observations as abrupt changes in enzyme functioning; anomalous temperature effects in membrane processes; unusual cell volume responses to temperature; dramatic changes in erythrocyte sedimentation rates; anomalies in chromosome aberration rates; pasteurization temperature; upper and lower thermal limits for microbial growth; multiple growth optima and minima and body temperatures of mammals and birds. PMID- 11728101 TI - Phase transitions and molecular motion in the cell. AB - The cytoplasm exhibits all of the signature characteristics of a gel. The thesis put forth here is that the cytoplasm's gel-like character is central to the generation of biological movement. In artificial gels, a common vehicle for generating movement is the polymer-gel phase-transition. By undergoing phase transition, gels produce motion of both solvent and solutes. It is argued that cells do the same. Three examples are given: the secretory system, the muscle contraction system and the biological streaming system. In each case it is shown that the characteristic motions may be created as proteins and water undergo transition from an expanded, hydrated state to a contracted, dehydrated state--or the reverse. These changes shift solutes and solvent in a characteristic way that depends on the respective organelle's structure. Phase-transitions are simple, powerful mechanisms that may be responsible for many, if not all, biological motions. PMID- 11728102 TI - Water and the cytoskeleton. AB - The diffusion of intracellular fluid and solutes is mainly limited by the density and the geometry of crossbridges between cytoskeletal polymers mediating the formation of an integrated cytoplasmic scaffold. Evidence for specific relationships between water and cytoskeletal polymers arises from the effect of heavy water on their polymerization process in vitro and on the cytoskeleton of living cells. The hydration of cytoskeletal subunits is modified through polymerization, a mechanism which may be involved in the direct contribution of the cytoskeleton to the osmotic properties of cells together with changes of hydration of polymers within networks. The dynamic properties of the hydration layer of cytoskeletal polymers may reflect the repetitive distribution of the surface charges of subunits within the polymer lattice, thus inducing a local and long range ordering of the diffusion flows of water and solutes inside polymer networks. The interactions between subunits in protofilaments and between protofilaments determine the specific viscoelastic properties of each type of polymer, regulated by associated proteins, and the mechanical properties of the cell through the formation of bundles and gels. Individual polymers are interconnected into dynamic networks through crossbridging by structural associated proteins and molecular motors, the activity of which involves cooperative interactions with the polymer lattice and likely the occurence of coordinated modifications of the hydration layer of the polymer surface. The cytoskeletal polymers are polyelectrolytes which constitute a large intracellular surface of condensed anionic charges and form a buffering structure for the sequestration of cations involved in the regulation of intracellular events. This property allows also the association of cytoplasmic enzymes and multimolecular complexes with the cytoskeleton, facilitating metabolic channelling and the localization of these complexes in specific subdomains of the cytoplasm. The consequences of interactions between membranes and the cytoskeleton in all cellular compartments range from the local immobilization and clustering of lipids and membrane proteins to the regulation of water and ion flows by the association of cytoskeletal subunits or polymers with transmembrane channels. The possibility that the polyelectrolyte properties of the cytoskeletal polymers contribute to the modulation of membrane potentials supports the hypothesis of a direct involvement of the cytoskeleton in intercellular communications. PMID- 11728103 TI - 1H-NMR studies of the intracellular water of skeletal muscle fibers under various physiological conditions. AB - Intracellular water of frog skeletal muscle fibers has been studied under various physiological conditions by use of the 1H-nuclear magnetic resonance (NMR) technique. 1H-NMR spectra of muscle fibers had single peaks derived from the intracellular water. The spectra changed in a characteristic fashion when the fiber axis was aligned at various angles relative to the magnetic field of the NMR magnet. Further, the relaxation rates of the 1H-NMR spectra changed depending on the water content of muscle fibers, and in association with contraction and rigor formation of muscle fibers. The obtained results indicate that the intracellular water of muscle fibers is structured and aligned along the myofilaments, and further that the state of the intracellular water changes with physiological conditions. PMID- 11728104 TI - Ontogenetic changes of water states and structural organization in growing kidney beans: parameter-imaging based on the diffusion measurements. AB - Physical states of cell-associated water in growing kidney beans was studied by NMR microscopy. 1H-NMR images indicated the changes of water amount in the individual tissues. The seed gradually reduced water and water in the tissues of the pod varied according to growth stages. Images that reflect diffusive motion provide different contrast from that of the proton images; signal intensity was maintained in the tissues containing small amounts of water and showing weak signal on the proton images. These images are related to the activity of metabolism and physiological functions. The parameter-images provided the structural information about cellular matrix, the barrier spacing of water compartments and the reduced permeability of cell-membranes surrounding the compartment, were created based on the restricted diffusion of cell-associated water. The properties of the water compartment changed with the stages of growth in individual tissues of the fruits. Two types of water compartments were detected; one had approximately 65 microm spacing and constant permeability (20 25%) throughout the stages of growth, and another had the smaller and variable spacing (approximately 35 microm), and a lower permeability. The former belongs to the seed and the mesocarp, the exocarp and the epidermis of the pod. While the latter characterizes the properties of the primary tissue in the pod, the endocarp, which increased with development of the pod then showed reduction with the decrease of water in the pod with the fruit growth. PMID- 11728105 TI - Total and structured water in cancer: an NMR experimental study of serum and tissues in DMBA-induced OF1 mice. AB - Total water and structured water (fraction of total water which remains unfrozen below the transition point from the semisolid to solid state) were characterized by 1H NMR relaxometry in the sera and tissues of 3 groups of 30 female mice (C, H and L) receiving a single administration of DMBA and different diets. Mice given the diet H, containing the highest proportion of saturated fatty acids and processed starch, and the lowest phytochemicals content, presented the highest tumor incidence (lymphoma). This allowed 3 subgroups to be defined: subnormal (SN), small (T+) and large tumor (T++). Spin-lattice relaxation times of total water (Tlobs) in the sera and tissues did not significantly differ between C, H and L groups, and SN, T+ and T++ subgroups. In T+ mice, a decrease in the relative amount of structured water was noticed in the serum, liver and heart, while changes in the temperature dependence of the Tl of structured water (Tlsw) were observed between -21 degrees C and -42 degrees C. These results suggest a moderate increase in the rotational mobility of structured water molecules in the serum and the heart, and a pronounced decrease in the liver. Likewise, the modification of the Tlsvv temperature dependence curve's shape tends to confirm the existence of important conformational changes in the macromolecular assemblies, which markedly affect the properties of structured water, especially in the earliest stage of cancer development. PMID- 11728107 TI - Reconstructive surgery using an artificial dermis (Integra): results with 39 grafts. AB - Integra was initially developed for the primary coverage of acute burns. It acts as a network for dermal reconstruction. An epidermal graft overlay is necessary after 3 weeks to achieve the in vivo reconstruction of a full-thickness skin equivalent. The quality of the functional and aesthetic results achieved led us to evaluate the potential of Integra in the treatment of burn scars and for general reconstructive surgery. We present a series of 31 patients who underwent Integra grafting for reconstructive surgery at a total of 39 operational sites. The average area grafted per procedure was 267 cm(2). Complications (silicone detachment, failure of the graft, haematoma) were observed in nine cases. The length of follow-up ranged from 0.5 year to 4 years. Two patients (two sites) were lost to follow-up; the final results in the remaining patients were considered to be good in 28 cases, average in six cases and poor in three cases. The disadvantages of using Integra in reconstructive surgery are the necessity of two operations, the risks of infection under the silicone layer, of the silicone becoming detached and of recurrence of contraction. On the other hand, Integra has many advantages including its immediate availability, the availability of large quantities, the simplicity and reliability of the technique, and the pliability and the cosmetic appearance of the resulting cover. In the light of these preliminary results, Integra appears as a new alternative to full-thickness skin grafting, skin expansion and even skin flaps for reconstructive surgery. PMID- 11728106 TI - Information forgotten or overlooked: fundamental flaws in the conventional view of the living cell. AB - Old ideas often persist long after sound evidence dictates otherwise. I attempt to report one such case in the life sciences, by pointing out what are perceived to be fundamental flaws or questions in conventional wisdom. It is my experience that much evidence not in support of the well accepted membrane pump view of the living cell has been overlooked, forgotten or even ignored. In presenting this idea, the evolution of our knowledge from the establishment of cellular and protoplasmic theory to the emergence of solution theory is presented. The universal hypothesis based on physical chemical principles is presented, followed by the advent of the membrane-situated energy-requiring pump. The experimental demonstration of an inadequate energy supply for the first pump is discussed, followed by a review of new evidence that calls to question the use of dilute solution theory in describing adequately cellular function. Finally, roles for cellular water are suggested to explain the cellular exclusion of sodium and to serve as a barometer for the healthy state. Within the context of a metaphor, I attempt to qualitatively embrace the physical findings. It is concluded that the mobility of water molecules may be considered to change with the progression of normal tissue to a state of disease. These changes in the mobility of water molecules are "fingerprinted" by changes in the molecular motion of the solids. PMID- 11728108 TI - Treatment of facial haemangiomas: the present status of surgery. AB - Although conservative management is usually proposed for haemangiomas occurring in infancy, the presence of these tumours on the face may result in severe complications and provide an indication for treatment. In this paper, we report 35 patients who underwent surgical treatment for facial haemangiomas. The series consists of 23 females and 12 males, ranging in age from 2.5 months to 35 years. In six patients early surgery, before the age of 2 years, was performed because of severe complications, including visual occlusion, repeated bleeding and distortion of adjacent structures. In 16 children surgical resection of haemangiomas was carried out between 2 and 5 years of age, before complete involution. In 13 patients persisting haemangiomas were surgically treated at an older age. The operative technique depended on the location and size of the lesion, and focused on resection of the tumour and reconstruction of the adjacent structures when necessary. The postoperative outcomes were very satisfactory. Early surgery is mandatory in the management of large periocular haemangiomas, to prevent secondary amblyopia, and proliferative labial tumours, which are prone to bleeding and cause difficulty while eating. Early surgical treatment is also recommended for nasal-tip haemangiomas, which regress very slowly and may result in severe distortion of the cartilaginous framework. In conclusion, facial haemangiomas causing functional disturbance or serious psychological distress deserve surgical excision before the age of expected spontaneous regression; surgery can provide active treatment with excellent results and minimal morbidity. PMID- 11728109 TI - Treatment of cicatricial lagophthalmos: very small orbicularis oculi muscle pedicled skin flap. AB - There are many methods for the correction of cicatricial lagophthalmos. Skin and soft tissue should be used to reconstruct the defect after release of tension. We used a very small orbicularis oculi muscle pedicled skin flap for the correction of mild to moderate degrees of cicatricial lagophthalmos in six cases. A small skin island flap from near the skin defect, pedicled on the orbicularis oculi muscle, was transposed to fill the defect. The length of the skin flap ranged from 5 mm to 11 mm. With a follow-up period ranging from 6 months to 18 months (mean: 11 months),we obtained satisfactory functional and aesthetic results. The eyelid closure was much improved and the discomfort was relieved. All the flaps survived without any healing problems. This small orbicularis oculi myocutaneous flap is very reliable and has a wide range of motion. This technique can be applied to eyelid-skin defects of various causes. PMID- 11728110 TI - The subscapular approach in head and neck reconstruction with the pedicled latissimus dorsi myocutaneous flap. AB - We have devised a new approach for transferring the pedicled latissimus dorsi myocutaneous flap to the head and neck region, and have applied it successfully in several patients. Our approach is rapid and increases the arc of rotation in order to cover defects of the lower lateral and posterior aspects of the head and neck. The vascular pedicle is protected between the scapula and the rib cage. We have used this approach in 15 patients over the last 9 years with good results. PMID- 11728111 TI - Histological examination of the capsules surrounding Trilucent breast implants. AB - Following a statement by the UK Medical Devices Agency (MDA), soya-oil-filled Trilucent implants for breast augmentation were withdrawn from sale in March 1999. The most recent report on the toxicity of the Trilucent implant suggests that one of the breakdown products of the filler is an aldehyde with an explicit risk of genotoxic and teratogenic effects. Explantation of all these prostheses was advised by the MDA in June 2000. No guidance regarding capsulectomy was put forward by any publication. The operation of capsulectomy confers an increased morbidity compared with simple prosthetic replacement. This study looks at the capsules of 18 consecutive patients who had received Trilucent implants for cosmetic breast augmentation, and who had undergone explantation and capsulectomy. The mean duration of implantation was 3 years (range: 2 years to 4 years 9 months). The capsules were examined histologically. The significant features of all the capsules included a florid foreign-body type reaction, synovial metaplasia, a villous hyperplasia and the presence of refractile material within the substance of the capsule. It is postulated that the refractile material is filler material that has bled through the wall of the intact Trilucent implant, and as such would represent a potential genotoxic hazard. Until such time as this potential hazard has been investigated fully and a conclusion reached, we recommend acceptance of the increased morbidity, and the performance of capsulectomies when explanting Trilucent implants. PMID- 11728112 TI - A study to evaluate primary dressings for the application of cultured keratinocytes. AB - Despite the recent improvements in cell culture and dermal regeneration methods, tissue engineering of skin has yet to receive widespread acceptance in the management of burn injuries. The reasons for this are complex and include not only the inherent costs of (particularly) setting up and running such a system but also the continuing difficulties in achieving successful engraftment of the neoepidermis. The latter has previously been addressed in a number of ways, including improving the recipient bed and using pre-confluent delivery systems to allow earlier application of cells to that wound bed. One area that has received little attention is that of the optimal wound dressing to use with this technology; the cells are very poorly attached at early time points, and, in this context, the traditional dressing of paraffin gauze has never been formally assessed in comparison with newer materials. Using a porcine acute wound chamber model, we performed a prospective randomised trial to assess four different wound dressings with reference to the amount of epidermal cover gained and the histological quality of the regenerated skin after 3 weeks. Out of the four materials tested, polyurethane foam (Allevyn) was superior histologically (although equal in take rate with paraffin gauze), whilst polythene sheet (Opsite) and silicone sheet were substantially inferior. We conclude that the traditional dressing used with this technology should be compared with polyurethane foam in a clinical trial. In the future, novel dressings should be formally tested against traditional methods before being adopted. PMID- 11728113 TI - Obstetric brachial plexus palsy: a comparison of the degree of recovery after repair of a C6 ventral root avulsion in newborn and adult sheep. AB - The C6 motor rootlets were avulsed from the spinal cord in six newborn lambs to simulate a birth lesion of the upper root of the brachial plexus. Six 1-year-old sheep were used for comparison, and treated in a similar manner. The injury was repaired immediately in each group using an autologous coaxial freeze-thawed skeletal muscle graft. The animals were allowed to recover for 1 year after the surgery. The C6 root was then examined electrophysiologically and morphologically. The results were compared with those obtained from a group of untreated intact 1-year-old sheep. The fibre and axon diameters and myelin sheath thickness were significantly different in the group repaired as lambs when compared with the group repaired at the age of 1 year. There was also a significantly increased maximum conduction velocity and a greater range of conduction velocities within the nerve in the lambs. Central motor latency was significantly slower in the sheep than in the lambs. These findings would suggest a greater potential for recovery in the lambs after brachial plexus root avulsion injuries. PMID- 11728114 TI - The H-flap: a useful flap for forehead reconstruction. AB - We have found that double opposing rectangular advancement flaps in the forehead may be easily performed under either local or general anaesthesia, and are associated with high patient satisfaction and low morbidity. We claim no originality for the mode of reconstruction, but highlight its broad spectrum of application and its popularity with junior plastic surgeons passing through our unit. We have found that the term 'H-flap' provides a readily communicated alternative to 'double opposing rectangular advancement flaps'. PMID- 11728115 TI - Cherubism and its charlatans. AB - Cherubism is a rare hereditary condition characterised by progressive cystic proliferation of the mandible and maxilla in childhood, followed by post-pubertal involution of the process and jaw remodelling in adulthood. Its name is derived from the cherubic appearance that results from the jaw hypertrophy. Here, we present the case of a young boy with cherubism, in the context of his pedigree, to illustrate the clinical characteristics and their variable expression. We also seek to distinguish cherubism from central giant cell granuloma and giant cell tumour of the jaws, with which it holds a false synonymity. PMID- 11728116 TI - Mandibular reconstruction using a vascularised fibula osteocutaneous flap in a patient with pyknodysostosis. AB - We present the first successful report of mandibular reconstruction using a free fibula osteocutaneous flap in a patient with pyknodysostosis. The patient had recurrent refractory osteomyelitis of the mandible, which was initially treated with resection and reconstruction using an AO plate. Recurrent fistula formation with loosening of the plate was treated by radical re-excision and reconstruction with a free fibula flap. The postoperative course was uneventful; at 14 months the bony union was almost complete, without resorption or atrophy of the grafted fibula on radiography. PMID- 11728117 TI - 'Clicking-brain syndrome': a curious cause of insomnia. AB - This case report describes a curious cause of insomnia. A 93-year-old woman presented to our follow-up clinic with the complaint of insomnia secondary to an audible click emanating from her skull. The site of loud biphasic-sound production corresponded to an area of the scalp where a squamous cell carcinoma had been removed 11 years previously. PMID- 11728118 TI - The use of a lymphatic bridge in the management of head and neck lymphoedema. AB - Severe progressive lymphoedema of the whole of the head and neck is almost always due to the combined effects of surgery and radiotherapy, compounded by repeated infections or recurrent tumour. The condition is difficult to control, and manual lymphatic drainage is the mainstay of management. We present a case of progressive lymphoedema following treatment for an occult squamous carcinoma of the head and neck, and describe the use of a lymphatic bridge to drain the facial tissue. PMID- 11728119 TI - Failure of bromocriptine therapy to control juvenile mammary hypertrophy. AB - Rapid massive breast hypertrophy occasionally occurs at the time of puberty or during pregnancy, with breast size eventually becoming burdensome or incapacitating to the patient. Pregnancy-related breast hypertrophy is often arrested or reversed by reducing serum prolactin levels with bromocriptine therapy. Unfortunately, breast enlargement in our 12-year-old patient with massive juvenile mammary hypertrophy was unaffected by bromocriptine therapy despite a reduction of her prolactin to normal levels. Two reduction mammaplasties followed by subcutaneous mastectomy were required to control breast hypertrophy. Breast-tissue hypersensitivity to prolactin appears to be a characteristic of pregnancy-related gigantomastia. Our pubertal patient with juvenile mammary hypertrophy failed to respond to bromocriptine therapy, so the aetiology of this syndrome may involve breast-tissue hypersensitivity to hormones other than prolactin. PMID- 11728120 TI - Two innovations of the star-flap technique for nipple reconstruction. AB - Two innovations of the star-flap technique for nipple reconstruction are described. A combination of the star-flap technique and a contralateral nipple graft is indicated in patients with a large nipple and small areola on the contralateral side. It provides sufficient volume for the new nipple and improves the shape of the donor nipple. A combination of the star-flap technique and a banked costal-cartilage graft offers better nipple contour and projection than the conventional star-flap technique. Preparation of the cartilage graft is easy and does not result in additional scarring; the nipple projection can be expected to be maintained over a long period. PMID- 11728121 TI - Anatomically aberrant forearm arteries: an absent radial artery with co-dominant median and ulnar arteries. AB - A radial-artery free flap was dissected from a traumatically amputated forearm. During dissection the arterial tree was found to be abnormal, with no radial artery. The arterial supply was based on co-dominant median and ulnar arteries. This description raises issues regarding anatomical vascular aberrations when planning forearm flaps. PMID- 11728122 TI - Recreation of the natal cleft by reverse Z-plasty. AB - The use of Z-plasties in the closure of elliptical or circular defects is not uncommon. We present a patient in whom a double Z-plasty was performed in order to close a sacral pressure sore. The patient returned 21 years later seeking aesthetic improvements of the resultant obliteration of the natal cleft and the soft tissue and skin redundancy in the central inferior aspect of the buttocks. We met this challenge with a practical solution. The triangular flaps of the Z plasties were returned to their original positions. This resulted in a natural shortening of the central portion of the buttock and the recreation of the natal cleft. The postoperative result was aesthetically pleasing. We report this unusual case and discuss the geometry behind the reversal of the Z-plasties as the solution to this problem. In addition, a novel interpretation of Z-plasties is presented. PMID- 11728123 TI - Breast implants and gel-filled sizers. PMID- 11728124 TI - Retraction suture for proximal interphalangeal joint exposure. PMID- 11728125 TI - Median forehead flaps for eyelid reconstruction. PMID- 11728126 TI - Multistage type III venous flap or 'pre-arterialisation of an arterialised venous flap'. PMID- 11728127 TI - 'Four-swab technique' for detecting bleeding following complicated breast reduction surgery. PMID- 11728131 TI - Intracellular stores maintain stable cytosolic Ca(2+) gradients in epithelial cells by active Ca(2+) redistribution. AB - A stable localized region of high calcium concentration near the plasma membrane has been postulated to exist as an outcome of prolonged calcium influx and to play a crucial role in regulation of cellular life. However, the mechanism supporting this phenomenon is a perplexing problem. We show here that a sustained localized region of high cytosolic Ca(2+) concentration is formed near the plasma membrane. Calcium influx, calcium uptake by intracellular stores and calcium release from the stores are essential for this phenomenon. Our results strongly suggest that the mechanism of formation of stable calcium gradient near the plasma membrane involves a process of active redistribution-uptake of entering calcium into intracellular stores and its release from the stores toward the plasma membrane. PMID- 11728132 TI - Calcium signaling mechanisms in dedifferentiated cardiac myocytes: comparison with neonatal and adult cardiomyocytes. AB - Our studies focused on calcium sparking and calcium transients in cultured adult rat cardiomyocytes and compared these findings to those in cultured neonatal and freshly isolated adult cardiomyocytes. Using deconvolution fluorescence microscopy and spec trophotometric image capture, sequence acquisitions were examined for calcium spark intensities, calcium concentrations and whether sparks gave rise to cell contraction events. Observations showed that the preparation of dedifferentiated cardiomyocytes resulted in stellate, neonatal-like cells that exhibited some aspects of calcium transient origination and proliferation similar to events seen in both neonatal and adult myocytes. Ryanodine treatment in freshly isolated adult myocytes blocked the calcium waves, indicating that calcium release at the level of the sarcoplasmic reticulum and t-tubule complex was the initiating factor, and this effect of ryanodine treatment was also seen in cultured-dedifferentiated adult myocytes. However, experiments revealed that in both neonatal and cultured adult myocytes, the inositol triphosphate pathway (IP3) was a major mechanism in the control of intracellular calcium concentrations. In neonatal myocytes, the nucleus and regions adjacent to the plasma membrane we re major sites of calcium release and flux. We conclude: (1) culturing of adult cardiomyocytes leads them to develop mechanisms of calcium homeostasis similar in some aspects to those seen in neonatal cardiomyocytes; (2) neonatal myocytes rely on both extracellular and nuclear calcium for contractile function; and (3) freshly isolated adult myocytes use sarcoplasmic reticulum calcium stores for the initiation of contractile function. PMID- 11728133 TI - Construction of a two-photon microscope for video-rate Ca(2+) imaging. AB - We describe the construction of a video-rate two-photon laser scanning microscope, compare its performance to a similar confocal microscope, and illustrate its use for imaging local Ca(2+) transients from cortical neurons in brain slices. Key features include the use of a Ti-sapphire femtosecond laser allowing continuous tuning over a wide (700-1000 nm) wavelength range, a resonant scanning mirror to permit frame acquisition at 30 Hz, and efficient wide-field fluorescence detection. Two-photon imaging provides compelling advantages over confocal microscopy in terms of improved imaging depth and reduced phototoxicity and photobleaching, but the high cost of commercial instruments has limited their widespread adoption. By constructing one's own system the expense is greatly reduced without sacrifice of performance, and the microscope can be more readily tailored to specific applications. PMID- 11728134 TI - Modes of secretagogue-induced [Ca(2+)](i) responses in individual chromaffin cells of the perfused rat adrenal medulla. AB - Chromaffin cells in the perfused rat adrenal medulla were loaded with indo-1 for confocal image analyses. Resting levels of [Ca(2+)](i) in chromaffin cells were similar and were stable with time. This is in contrast to the situation in isolated rat chromaffin cells, in which spontaneous oscillations of [Ca(2+)](i) are known to occur. When chromaffin cells were stimulated for 3-4 min by high K(+) or nicotine, [Ca(2+)](i) increased to a peak in 20-30 s and then declined rather smoothly. In contrast, chromaffin cells stimulated by muscarine or low pH (6.5) commonly exhibited irregular oscillations in [Ca(2+)](i). This provides additional evidence supporting the previous claim that muscarine and low pH evoke catecholamine secretion using partly shared mechanisms. Although muscarine and low pH were speculated to produce weaker responses in noradrenaline-secreting cells due to their selective stimulation of adrenaline secretion, no clear indications for segregation of cell types from [Ca(2+)](i) responses to these stimulants were found. The perfused adrenal medulla loaded with Indo-1 was also employed for simultaneously monitoring integrated changes in [Ca(2+)](i)(Ca responses) by conventional microfluorometry and in catecholamine secretion from a whole medulla (secretory responses). When the profiles of secretory responses were approximated by the kth power of the profiles of Ca responses, the k-values were estimated to be 2.2 and 2.3 for high-K(+)- and nicotine-elicited responses, respectively, whereas a k-value of 1.4 was obtained for both muscarine- and low pH-elicited responses. An analysis showed that the significant difference in the k-value with these two classes of stimulants is accounted for by the stimulant dependent patterns of [Ca(2+)](i) responses found in confocal image analysis. PMID- 11728135 TI - Intracellular calcium in the isolated rat liver: correlation to glucose release, K(+) balance and bile flow. AB - This study correlates whole organ measurements of intracellular calcium concentration ([Ca(2+)](i)) with hormone-induced (epinephrine, vasopressin) changes of liver functions (glucose release, K(+) balance and bile flow). [Ca(2+)](i) was measured in the isolated perfused rat liver using the sensor Fura 2 and applying liver surface fluorescence spectroscopy. The technique was improved by (i) minimizing biliary elimination of the sensor by employing a rat strain deficient in canalicular organic anion transport (TR(-) mutation) and (ii) by correcting for changes of interfering intrinsic organ fluorescence that was shown to depend on the oxidation-reduction state (NAD(P)H content) of the organ. Epinephrine (50 nM) elicits an instantaneous peak rise of [Ca(2+)](i) to approx. 400 nM, followed by a sustained elevation that depends on the presence of extracellular Ca(2+). The rise of [Ca(2+)](i) coincides with initiation of glucose release, transient K(+) uptake, and transient stimulation of bile flow. Vasopressin (2 nM) exerts qualitatively similar effects. The transient rise of bile flow is attributed to Ca(2+)-mediated contraction of the pericanalicular actin-myosin web of hepatocytes. PMID- 11728136 TI - The nucleus together with the cytosol generates patterns of specific cellular calcium signatures in tobacco suspension culture cells. AB - Plant cell suspension cultures respond to osmotic changes by alterations in levels of free cellular calcium. Using the aequorin recombinant method, we have measured the spatial and temporal characteristics of calcium signatures in the nucleus and the cytosol of BY-2 tobacco suspension cells challenged with hypo- or hyper-osmotic shock. We show here that the nuclear compartment contributes together with the cytosol to produce calcium signal patterns that discriminate hypo- from hyper-osmotic treatments, i.e. turgor from tension. We also demonstrate that calcium responses in the nucleus and the cytosol are differentially modulated by the strength and the nature of hyper-osmotic treatments. We conclude that qualitative and quantitative changes in the parameters of an external stimulus such as osmotic changes are converted into calcium signatures, distinctive in their temporal and subcellular characteristics, involving both the nucleus and the cytosol. Our results illustrate the versatility of calcium signaling in plant cells. In addition to the physiological 'address' of the cell, the compartmentation of the calcium signal is probably an important parameter in encoding response specificity. PMID- 11728137 TI - Mitochondrial modulation of calcium signaling at the initiation of development. AB - Fertilization triggers cytosolic Ca(2+) oscillations that activate mammalian eggs and initiate development. Extensive evidence demonstrates that Ca(2+) is released from endoplasmic reticulum stores; however, less is known about how the increased Ca(2+) is restored to its resting level, forming the Ca(2+) oscillations. We investigated whether mitochondria also play a role in activation-associated Ca(2+) signaling. Mitochondrial dysfunction induced by the mitochondrial uncoupler FCCP or antimycin A disrupted cytosolic Ca(2+) oscillations, resulting in sustained increase in cytosolic Ca(2+), followed by apoptotic cell death. This suggests that functional mitochondria may participate in sequestering the released Ca(2+), contributing to cytosolic Ca(2+) oscillations and preventing cell death. By centrifugation, mouse eggs were stratified and separated into fractions containing both endoplasmic reticulum and mitochondria and fractions containing endoplasmic reticulum with no mitochondria. The former showed Ca(2+) oscillations by activation, whereas the latter exhibited sustained elevation in cytosolic Ca(2+) but no Ca(2+) oscillations, suggesting that mitochondria take up released cytosolic Ca(2+). Further, using Rhod-2 for detection of mitochondrial Ca(2+), we found that mitochondria exhibited Ca(2+) oscillations, the frequency of which was not different from that of cytosolic Ca(2+) oscillations, indicating that mitochondria are involved in Ca(2+) signaling during egg activation. Therefore, we propose that mitochondria play a crucial role in Ca(2+) signaling that mediates egg activation and development, and apoptotic cell death. PMID- 11728138 TI - Gene polymorphisms of pro- (or anti-) inflammatory cytokines and vascular disease. PMID- 11728139 TI - Caeteris paribus - all things being equal. PMID- 11728140 TI - Fibrinogen polymorphisms and disease. PMID- 11728141 TI - Refractory heart failure - drugs and devices. PMID- 11728142 TI - Dealing with biological variation in the Brugada syndrome. PMID- 11728143 TI - Platelets and atrial fibrillation. PMID- 11728144 TI - The interleukin-6 -174 G/C promoter polymorphism is associated with risk of coronary heart disease and systolic blood pressure in healthy men. AB - AIMS: Inflammation is a key component of coronary heart disease, and genes coding for cytokines are candidates for predisposing to coronary heart disease risk. We have examined the effect of two polymorphisms (-174G>C and -572G>C) in the promoter of the interleukin-6 (IL-6) gene on risk of coronary heart disease, and on intermediate risk traits including fibrinogen and systolic blood pressure, in 2751 middle-aged healthy U.K. men. RESULTS: The -174C allele (frequency 0.43, 95% CI 0.42-0.44) was not associated with significant effects on fibrinogen levels, but was associated with a significantly (P=0.007) higher systolic blood pressure (mean mmHg (95% CI): GG=135.5 (134.3-136.7); GC=137.9 (136.9-138.9); CC= 138.0 (136.3-139.8)). This effect was of similar magnitude in smokers and non-smokers, and was greater in men in the top two tertiles of body mass index (>24.86 kg x m( 2)) than in those in the bottom tertile. Compared to those with the genotype GG, men carrying the -174C allele had a relative risk of coronary heart disease of 1.54 (95% CI 1.0-2.23, P=0.048) and this effect was greatest in smokers (compared to GG non-smokers, RR 2.66, CI 1.64-4.32). These effects remained statistically significant after adjusting for classical risk factors including blood pressure (P=0.04). The -572C allele (frequency 0.05, 0.04-0.06) was not associated with a significant effect on blood pressure, fibrinogen or relative risk of coronary heart disease. In a subset of the genotyped men (n=494), carriers of the -174C allele had higher levels of C-reactive protein than non-carriers. CONCLUSIONS: These data confirm the importance of the inflammatory system in the development of coronary heart disease. They suggest that, at least in part, the effect of the IL-6 -174G>C polymorphism on blood pressure is likely to be operating through inflammatory mechanisms, but the genotype effect on coronary heart disease risk is largely unexplained by its effect on blood pressure. The molecular mechanisms whereby genetically determined differences in plasma levels of IL-6 are having these effects remain to be determined. PMID- 11728145 TI - Incidence and predictors of bleeding events after fibrinolytic therapy with fibrin-specific agents: a comparison of TNK-tPA and rt-PA. AB - BACKGROUND: Fibrinolytic therapy increases the risk of bleeding events. TNK-tPA (tenecteplase) is a variant of rt-PA with greater fibrin specificity and reduced plasma clearance that can be given as a single bolus. We compared the incidence and predictors of bleeding events after treatment with TNK-tPA and rt-PA. METHODS AND RESULTS: In the Assessment of the Safety and Efficacy of a New Thrombolytic (ASSENT)-2 trial, 16 949 patients with acute myocardial infarction were randomly assigned a single weight-adjusted bolus of TNK-tPA or a 90-min infusion of rt-PA. A total of 4.66% of patients in the TNK-tPA group experienced major non-cerebral bleeding, in comparison with 5.94% in the rt-PA group (P=0.0002). This lower rate was associated with a significant reduction in the need for blood transfusion (4.25% vs 5.49%, P=0.0003) and was consistent across subgroups. Independent risk factors for major bleeding were older age, female gender, lower body weight, enrolment in the U.S.A. and a diastolic blood pressure <70 mmHg. Females at high risk (age >75 years and body weight <67 kg) were less likely to have major bleeding when treated with TNK-tPA even after other risk factors were taken into account. A total of 0.93% of patients in the TNK-tPA and 0.94% of patients in the rt-PA group experienced an intracranial haemorrhage. Female patients >75 years of age who weighed <67 kg tended to have lower rates of intracranial haemorrhage when treated with TNK-tPA (3/264, 1.14% vs 8/265, 3.02%). CONCLUSIONS: The increased fibrin specificity and single bolus administration of TNK-tPA do not increase the risk of intracranial haemorrhage but are associated with less non cerebral bleeding, especially amongst high-risk patients. PMID- 11728146 TI - Mutation in the promoter region of the beta-fibrinogen gene and the risk of future myocardial infarction, stroke and venous thrombosis. AB - AIM: Polymorphisms in the promoter region of the beta-fibrinogen gene are associated with increased plasma fibrinogen levels. We investigated whether the distribution of the C148T polymorphism is associated with an increase in cardiovascular risk. METHODS AND RESULTS: In a nested case-control design, the distribution of the C148T polymorphism was investigated among 751 participants in the Physicians' Health Study who subsequently developed myocardial infarction, stroke or venous thromboembolism (cases) and among 751 age- and smoking-matched controls over follow-up of 8.6 years. Frequency of the T allele was similar among men who had myocardial infarction (22.7%, P=0.5), stroke (18.4%, P=0.2) or venous thromboembolism (17.0%, P=0.1) compared with those with no cardiovascular events (21.5%). The relative risk for any vascular event among men homozygous or heterozygous for the T allele compared with men homozygous for the C allele was 0.94 (95% CI 0.76-1.16). We found no evidence of an association between the T allele and myocardial infarction (relative risk 1.06; 95% CI 0.82-1.36), stroke (0.87, 0.63-1.21) or venous thromboembolism (0.75; 0.51-1.08). Analysis adjusted for aspirin use and traditional cardiovascular risk factors had no significant effect on these findings. CONCLUSION: In a large prospective cohort, carriage of the T allele for the C148T mutation in the beta-fibrinogen promoter gene was not associated with an increased subsequent risk of cardiovascular events. PMID- 11728147 TI - Combination treatment in stable effort angina using trimetazidine and metoprolol: results of a randomized, double-blind, multicentre study (TRIMPOL II). TRIMetazidine in POLand. AB - AIMS: To assess the antiischaemic efficacy and tolerability of the metabolic agent trimetazidine in combination with metoprolol in patients with stable effort angina. METHODS: This was a randomized, multicentre, double-blind, placebo controlled parallel group study. A total of 426 male and female patients with stable, effort-induced angina and documented coronary artery disease received either placebo or trimetazidine 20 mg three times daily in addition to metoprolol 50 mg twice daily. Treadmill exercise tests were performed at weeks (-1), 0, 4 and 12. RESULTS: After 12 weeks, there were significantly greater improvements in the metoprolol + trimetazidine group than in the metoprolol + placebo group in: time to 1 mm ST segment depression, total workload, time to onset of angina, maximum ST segment depression, mean weekly number of angina attacks, mean weekly nitrate consumption, and grade of anginal pain. There was no evidence of any development of tolerance to trimetazidine. The tolerability of trimetazidine was excellent. CONCLUSIONS: Therapy with trimetazidine plus metoprolol produced significant improvements in exercise stress tests and the symptoms of angina relative to metoprolol alone. With its metabolic effect, devoid of any haemodynamic action, trimetazidine is useful for combination therapy in patients with stable angina insufficiently controlled by monotherapy with a beta-blocker. PMID- 11728148 TI - Ventricular assist device in severe heart failure: effects on cytokines, complement and body weight. AB - AIMS: Inflammatory and immune activation and body wasting are important features of end-stage chronic heart failure. It is not known whether restoration of cardiac output by assist device implantation can improve these abnormalities. METHODS: We studied 48 patients (39 males; age 45+/-2 years) with NYHA class IV heart failure. All patients underwent ventricular assist device implantation for end-stage heart failure as a bridge to cardiac transplantation. Plasma levels of tumour necrosis factor alpha, and its receptors, interleukin-6, elastase, activated complement, and soluble CD14 receptors were measured at the time of operation and in survivors at 1 week (n=46), 40 days (n=35) and 90 days (n=26). Follow-up was for a minimum of 1 year. RESULTS: One-year survival was 35% (95% CI: 22-49%). Body mass index was the only predictor of survival (body mass index >25 (n=16); survival 63 (39-86) %; body mass index <25 (n=32); survival 22 (7.5 36) %: P=0.003). Tumour necrosis factor alpha fell from 9.66+/-1.33 pg x ml(-1) to 4.2+/-1.0 at 1 week (P=0.008), but returned to pre-operative levels at 90 days. Interleukin-6, activated complement and elastase fell progressively to 40 days, but were rising at 90 days. There was no change in tumour necrosis factor receptor. There was a gradual rise in CD14 (3.99+/-0.15 microg x ml(-1) at baseline, 5.02+/-0.39 at 90 days, P=0.006). After surgery, body weight fell from 80+/-2 to 73+/-2 kg by 1 month (P<0.001) and to 72+/-2 kg at 90 days. CONCLUSIONS: Ventricular assist device implantation results in a short-term fall in tumour necrosis factor alpha and interleukin-6, but no change in CD14 or tumour necrosis factor receptor, suggesting that the pathophysiological process resulting in inflammation was not altered by left ventricular assist device implantation. Low body mass index is related to poor outcome after assist device implantation, and no weight gain. PMID- 11728149 TI - Frequency and clinical characteristics of dilated cardiomyopathy caused by desmin gene mutation in a Japanese population. AB - AIMS: Dilated cardiomyopathy is partly caused by a mutation of some cytoskeletal or nuclear envelope proteins. It has been confirmed recently that a missense mutation of the gene encoding desmin, a cytoskeletal protein, can cause dilated cardiomyopathy. This study was aimed at elucidating the frequency and clinical characteristics of dilated cardiomyopathy caused by desmin mutation. METHODS AND RESULTS: We examined 265 Japanese patients with dilated cardiomyopathy (217 sporadic cases and 48 probands of familial dilated cardiomyopathy). The exon 8 of the desmin gene, the critical region for the pathogenesis of dilated cardiomyopathy, was analysed by polymerase chain reaction, single-strand conformation polymorphism and sequencing. The same missense mutation (Ile451Met) as reported previously was detected in three patients (1.1%). All these patients were male and sporadic, and more likely to be accompanied by characteristics such as younger age at diagnosis, lower fractional shortening and ejection fraction than each mean value of sporadic cases. The chronological changes in cardiac function were inconsistent in the three patients. CONCLUSION: The missense mutation (Ile451Met) of the desmin gene can be the genetic cause of dilated cardiomyopathy, although with very low frequency. The ages at diagnosis were younger and the cardiac function had deteriorated further than general cases of sporadic dilated cardiomyopathy. PMID- 11728150 TI - New electrocardiographic leads and the procainamide test for the detection of the Brugada sign in sudden unexplained death syndrome survivors and their relatives. AB - AIMS: Sudden unexplained death syndrome occurs in previously healthy South-east Asian young adults without any structural cause of death. The common electrocardiographic (ECG) change in sudden unexplained death syndrome survivors is right bundle branch block and ST elevations in leads V(1) to V(3), which are similar to the ECG pattern in the Brugada syndrome (Brugada sign). It is difficult to diagnose the Brugada sign with the 12-lead ECG in sudden unexplained death syndrome survivors and their family members because the ECG could be transiently normalized. We proposed using the higher intercostal space V(1) to V(3) lead ECG, together with procainamide to detect the Brugada sign. METHODS AND RESULTS: Among 20 ventricular fibrillation cardiac arrest patients, 13 sudden unexplained death syndrome survivors and their relatives (n=88) were studied using the single standard 12-lead ECG and the new six higher intercostal space V(1) to V(3) lead ECG (-V(1) to -V(3) and -2V(1) to -2V(3)). Ten sudden unexplained death syndrome survivors and relatives (n=48) who had a normalized ECG were also infused with procainamide (10 mg x kg(-1)i.v.) to unmask the Brugada sign and both ECG methods were recorded. Forty healthy individuals and 13 spouses served as the control group. Prior to the procainamide infusion, the Brugada sign could be detected in nine sudden unexplained death syndrome survivors (69.2%) and three (3.4%) relatives with the standard ECG and in 12 (92.3%) and nine (10.2%) with the new six-lead ECG. After the procainamide infusion, the Brugada sign could be demonstrated in seven sudden unexplained death syndrome survivors (70%) and seven (14.6%) relatives with the standard ECG and in nine (90%) (P=0.26) and 23 (47.9%) (P=0.0004) with the new six-lead ECG, respectively. All the controls were negative for the Brugada sign. CONCLUSIONS: Our data suggest that the new higher intercostal space lead ECG, with or without the procainamide test is helpful in detecting the Brugada sign in sudden unexplained death syndrome survivors and their relatives. PMID- 11728153 TI - Pneumocystis carinii: genetic diversity and cell biology. AB - As an important opportunistic pulmonary pathogen, Pneumocystis carinii has been the focus of extensive research over the decades. The use of laboratory animal models has permitted a detailed understanding of the host-parasite interaction but an understanding of the basic biology of P. carinii has lagged due in large part to the inability of the organism to grow well in culture and to the lack of a tractable genetic system. Molecular techniques have demonstrated extensive heterogeneity among P. carinii organisms isolated from different host species. Characterization of the genes and genomes of the Pneumocystis family has supported the notion that the family comprises different species rather than strains within the genus Pneumocystis and contributed to the understanding of the pathophysiology of infection. Many of the technical obstacles in the study of the organisms have been overcome in the past decade and the pace of research into the basic biology of the organism has accelerated. Biochemical pathways have been inferred from the presence of key enzyme activities or gene sequences, and attempts to dissect cellular pathways have been initiated. The Pneumocystis genome project promises to be a rich source of information with regard to the functional activity of the organism and the presence of specific biochemical pathways. These advances in our understanding of the biology of this organism should provide for future studies leading to the control of this opportunistic pathogen. PMID- 11728154 TI - Characterization of four clustered and coregulated genes associated with fumonisin biosynthesis in Fusarium verticillioides. AB - Fumonisins are mycotoxins that cause several fatal animal diseases, including cancer in rats and mice. These toxins are produced by several Fusarium species, including the maize pathogen Fusarium verticillioides, and can accumulate in maize infected with the fungus. We have identified four F. verticillioides genes (FUM6, FUM7, FUM8, and FUM9) adjacent to FUM5, a previously identified polyketide synthase gene that is required for fumonisin biosynthesis. Gene disruption analysis revealed that FUM6 and FUM8 are required for fumonisin production and Northern blot analysis revealed that expression of all four recently identified genes is correlated with fumonisin production. Nucleotide sequence analysis indicated that the predicted FUM6 translation product is most similar to cytochrome P450 monooxygenase-P450 reductase fusion proteins and the predicted products of FUM7, FUM8, and FUM9 are most similar to type III alcohol dehydrogenases, class-II alpha-aminotransferases, and dioxygenases, respectively. Together, these data are consistent with FUM5 through FUM9 being part of a fumonisin biosynthetic gene cluster in F. verticillioides. PMID- 11728155 TI - Mechanics of solid tissue invasion by the mammalian pathogen Pythium insidiosum. AB - The relative significance of mechanical penetration versus the action of substrate-degrading enzymes during solid tissue invasion has not been established for any fungal disease. Pythium insidiosum is an oomycete fungus (or stramenopile) that causes a rare, but potentially lethal infection in humans and other mammalian hosts. Experiments with miniature strain gauges showed that single hyphal apices of this pathogen exert forces of up to 6.9 microN, corresponding to maximum pressures of 0.3 microN microm(-2) or MPa. Samples of cutaneous and subcutaneous tissue from fresh human cadavers displayed a mean strength (resistance to needle puncture) of 24 microN microm(-2), and a mean pressure of 30 microN microm(-2) was necessary to penetrate skin strips from slaughtered horses. These experiments demonstrate that P. insidiosum does not exert sufficient pressure to penetrate undamaged skin by mechanics alone, but must effect a decisive reduction in tissue strength by proteinase secretion. PMID- 11728156 TI - Potential role of transposable elements in the rapid reorganization of the Fusarium oxysporum genome. AB - The activity of several families of transposable elements (TEs) in the genome of Fusarium oxysporum represents a potential source of karyotypic instability. We investigated transposon-mediated chromosome rearrangements by analyzing the karyotypes of a set of strains in which transposition events had occurred. We uncovered exceptional electrophoretic karyotype (EK) variability, in both number and size of chromosomal bands. We showed that EK differences result from chromosomal translocations, large deletions, and even more complex rearrangements. We also revealed many duplicated chromosomal regions. By following transposition of two elements and analyzing the distribution of different families of TEs on whole chromosomes, we find (i) no evidence of chromosomal breakages induced by transposition, (ii) a clustering of TEs in some regions, and (iii) a correlation between the high level of chromosomal polymorphism and the concentration of TEs. These results suggest that chromosome length polymorphisms likely result from ectopic recombination between TEs that can serve as substrates for these changes. PMID- 11728157 TI - Characterization of the gdhA gene from the phytopathogen Botrytis cinerea. AB - A 3.48-kb DNA region containing the gdhA gene, which codifies the NADP-dependent glutamate dehydrogenase enzyme from Botrytis cinerea, has been cloned and characterized. A fragment of 2351 nucleotides was sequenced and found to contain an ORF of 1350 bp that encodes a protein of 450 amino acids. The gene, containing two introns that showed polymorphic size between them, was located by pulsed field gel electrophoresis in chromosome X in seven strains, which were isolated from several hosts and had different levels of pathogenesis. The protein was similar to the gdhA of various other organisms, with nine highly conserved motifs that included the known active site sequence. The cloned gene was proven to be functional since it complemented two different Aspergillus nidulans gdhA mutants, restoring high levels of NADP-dependent glutamate dehydrogenase activity to the transformants. gdhA was transcribed as a monocistronic transcript of 1.7 kb starting at an A or a T, located 40 or 47 bp, respectively, upstream from the initial ATG codon of the ORF. Transcription levels of the gdhA gene were high during the rapid growth phase. Very high expression levels of the gdhA gene were observed in media with asparagine as the nitrogen source, whereas glutamic acid repressed transcription of the gdhA gene. Similarly high levels of gdhA gene transcription were observed in media with acetate as the carbon source, while glycerol strongly repressed gdhA gene transcription. These results indicate that expression of the gdhA gene is subject to strong nitrogen and carbon regulation at the transcriptional level. PMID- 11728158 TI - Expression of recombinant aequorin as an intracellular calcium reporter in the phytopathogenic fungus Phyllosticta ampelicida. AB - Conidia of Phyllosticta ampelicida germinate only after they have made contact with a substratum. Previous work has shown that external free calcium must be available to the spore for germination to be initiated. Transgenic strains of P. ampelicida expressing apo-aequorin, a calcium-sensitive luminescent protein, were developed to monitor cytoplasmic free Ca(2+) ([Ca(2+)]c). Transformants were verified by PCR and Southern hybridization. Apo-aequorin production was quantified for each of 21 transformants. The transformant that emitted the most light per unit of protein was found to contain 0.59 mg apo-aequorin/g total protein. To ascertain the feasibility of aequorin-based [Ca(2+)]c quantification, [Ca(2+)]c changes were measured in mycelia during various physiologically perturbing treatments: exposure to high concentrations of external Ca(2+), hypoosmotic shock, and mechanical perturbation. This is the first report of a plant pathogenic fungus for which aequorin-based Ca(2+) measurement protocols have been developed. PMID- 11728159 TI - Aspergillus nidulans alpha-1,3 glucanase (mutanase), mutA, is expressed during sexual development and mobilizes mutan. AB - We established a subtractive cDNA library of Aspergillus nidulans to identify differentially expressed genes during sexual development. One of the clones displayed homology to fungal alpha-1,3 glucanases (mutanase). Since alpha-1,3 glucan is considered the main reserve material accumulated during vegetative growth as a cell wall component and consumed during sexual development, we analyzed this gene in detail. The gene, mutA, is disrupted by three introns and encodes a putative protein of 48 kDa molecular mass with a signal peptide for secretion at the N terminus. The deduced protein displays amino acids 24-42% identical to mutanases of other fungi. A proposed mutan binding domain characterized in, e.g., Penicillium is not present in A. nidulans. Mutanase transcript and GFP reporter analysis in A. nidulans revealed specific induction of the gene during sexual development in Hulle cells. To study the role of mutA during sexual differentiation, we constructed a mutA deletion strain. Although degradation of mutan was affected in this strain, it was still able to form cleistothecia at a number similar to that of wildtype. These results suggest that additional carbon sources are available during sexual development. PMID- 11728161 TI - Opioid effects on glucose and eicosanoid metabolism in isolated uterus of ovariectomized and non-ovariectomized restricted diet rats. AB - The effect of a 25-day restricted diet (50% of the normal food intake) on uterine glucose metabolism of ovariectomized (25 days) and non-ovariectomized rats, was studied. Underfeeding reduces (14)CO(2) production from U(14)C-glucose in intact animal. However, in spayed rats, results are the opposite. In intact rats receiving a low food intake, the effect of the addition to the KRB medium of various agonist opioids, was studied. Dinorphin A did not bring about any change. On the other hand, beta endorphin increased glucose metabolism. Also, the addition of Dago and Dadle increased (14)CO(2) production, while their corresponding specific blockers, beta-FNA and Naltrindole, reversed it. Ovariectomized rats subjected to food restriction are not affected by opioid agonists. In vitro morphine, like endogenous opioids, increased (14)CO(2) in intact restricted diet rats. Arachidonic acid metabolism in these rats show that underfeeding brings about a decrease in PGF(2 alpha) and PGE(2), but the addition of morphine does not alter this situation, for which eicosanoids metabolites are not related to the effect of morphine. The morphine effect was not altered by naloxone. The subcutaneous injection of morphine increased glucose metabolism in intact underfed animals, while naloxone reduced (14)CO(2) in spayed rats subjected to underfeeding. It can be concluded that uteri from ovariectomized rats receiving a restricted diet are influenced by a mechanism of upregulation related to endogenous opioids. These likely originate in other tissues, and so prevent us from seeing the morphine effect. PMID- 11728162 TI - Prostaglandin E(2) modulation of vascular endothelial growth factor production in murine macrophages. AB - We have previously shown that dietary (n-3) fatty acids decrease mammary tumor vascularization and PGE(2) production. One possible mechanism may be the modulation of vascular endothelial growth factor (VEGF) production by PGE(2). Macrophages are major producers of VEGF, and thus we assessed the role of PGE(2) in vitro and in vivo on their VEGF production. When added to macrophages, pharmacological (10(-7) M) but not physiological (10(-9) to 10(-11) M) concentrations of PGE(2) increased VEGF mRNA and protein levels. That increased expression was relatively rapid and sustained up to 8 hrs, but declined by 24 hrs. Similarly, dibutryl cAMP increased production of VEGF protein which was completely inhibited by H89. Addition of cAMP-elevating agents further potentiated the production of VEGF by PGE(2). Next, (n-3) and (n-6) fatty acids were added to macrophages in vitro or provided in the diet. Macrophages of mice fed safflower oil (n-6) had 2- to 4-fold greater copy number of VEGF transcripts after lipopolysaccarhide (LPS) stimulation compared to fish oil (n-3). A decreasing trend was seen in LPS-induced VEGF secretion from macrophages in vitro after docosahexaenoic acid or eicosapentaenoic acid incubation compared to arachidonic acid. While pharmacological concentrations of PGE(2) modulate VEGF expression, physiological alterations did not alter VEGF protein production by macrophages. PMID- 11728163 TI - Increased synthesis of thromboxane A(2) and expression of procoagulant activity by monocytes in response to arachidonic acid in diabetes mellitus. AB - Thromboxane A(2) (TXA(2)) synthesis and expression of procoagulant activity (PCA) were investigated in mononuclear cells and monocytes prepared from a control and a Type 2 diabetic group. Monocytes from the diabetic group produced 2.10+/-0.81 ng of TXB(2)/5 x 10(5) monocytes compared to 1.26+/-0.43 ng/5 x 10(5) monocytes by the control group (P<0.01, n=11) when incubated in autologous plasma containing arachidonic acid (200 microg/ml). When monocytes were incubated in buffer containing arachidonic acid (20 microg/ml), cells from the diabetic group produced 1.65+/-0.68 ng of TXB(2)/5 x 10(5) monocytes compared to 1.07+/-0.31 ng/5 x 10(5) monocytes by the control group (P<0.02, n=12). Expression of PCA was examined in mononuclear cell preparations. Basal and maximally stimulated PCA with lipopolysaccharide (4.2 microg/ml) were not different between control and diabetic groups. However, arachidonic acid induced a four-fold (P<0.001) increase in PCA in the diabetic group. This activity was characterized as tissue factor. Increased synthesis of TXA(2) and expression of PCA may potentiate thrombosis and increase fibrin deposition, events that play primary roles in the development of vascular disease. PMID- 11728164 TI - Sphingosine-1-phosphate effects on PKC isoform expression in human osteoblastic cells. AB - Sphingosine-1-phosphate (S1P) has been shown to participate in the proliferative process in human osteoblasts.(1) The mitogenic effect of S1P has been postulated to involve two signaling pathways, the Gi linked protein receptor pathway and the PKC pathway. To define the possible role of PKC isoforms in osteoblastic cell proliferation, the effects of S1P on PKC isoform expression was determined. While PKC lambda was minimally detected, the isoforms alpha, delta and iota were all found to be highly expressed by the human osteoblast. In human osteoblastic cells, S1P induced a 25% increase in the expression of PKC alpha and approximately a 30% increase in the expression of PKC iota. S1P did not have an effect on PKC delta expression. Pretreatment with pertussis toxin (PT) led to an inhibition of the observed S1P effects on the expression of the alpha and iota isoforms. PMID- 11728165 TI - Fatty acid composition of the milk lipids of Nepalese women: correlation between fatty acid composition of serum phospholipids and melting point. AB - Milk was collected from 36 Nepalese women, 15 to 32 years of age, in order to investigate relationships between the proportions of intermediate chain-length (C10-C14) fatty acids and critical n-3 and n-6 polyunsaturated fatty acids in the milk lipids they were producing. Serum was also obtained from these lactating women and the fatty acid composition of their serum phospholipid fraction was determined and compared with that of the corresponding milk lipid fraction. Compared to women in technologically advanced parts of the world, the serum phospholipids of the Nepalese women contained nutritionally adequate proportions of linoleic acid (LA) (16.8%), alpha-linolenic acid (ALA) (0.53%), arachidonic acid (AA) (5.69%), and docosahexaenoic acid (DHA) (1.42%). However, although the milk lipids contained adequate proportions of ALA (1.81%), AA (0.43%), and DHA (0.23%), the lipids contained low to moderate percentages of LA (mean, 9.05%). Positive correlations were observed between the proportions of AA (P=0.001, r=0.50) and ALA (P=0.03, r=0.36) in the serum phospholipids and milk lipids of the women. As the proportion of C10-Cl4 fatty acids in the milk lipids increased from 10% to 40%, there was preferential retention of three critical n-3 and n-6 fatty acids (ALA, AA, and DHA) at the expense of two relatively abundant nonessential fatty acids, namely stearic acid and oleic acid. In addition, using fatty acid melting point data and the mol fraction of the 9 most abundant fatty acids in the milk, we estimated the mean melting point (MMP) of the milk lipids of the Nepalese women. The MMPs ranged from 29.3 to 40.5 degrees C (median, 35.5 degrees C). These results indicate that: 1) the levels of AA and ALA in the blood of lactating mothers influence the levels of these fatty acids in the milk they produce; 2) when the mammary gland produces a milk that is rich in C10-Cl4 fatty acids, it somehow regulates triglyceride synthesis in such a way as to ensure that the milk will provide the exclusively breast-fed infant with the amounts of the critical n-3 and n-6 fatty acids it requires for normal growth and development; and 3) the melting point of the milk lipid fraction is determined mainly by the mol % of the intermediate chain-length (C10-C14) fatty acids, oleic acid, linoleic acid, and alpha-linolenic acid. PMID- 11728166 TI - Endothelin involvement in respiratory centre activity. AB - To evaluate the role of endothelin (ET) in respiratory homeostasis we studied the effects of the ET(A) and ET(B) receptor blocking agent bosentan on respiratory mechanics and control in seven anaesthetised spontaneously breathing pigs, for 180 min after single bolus administration (20 mg/kg i.v.). The results show that the block of ET receptors induced a significant increase in compliance and decrease in resistance of the respiratory system, entailing a significant reduction of diaphragmatic electromyographic activity, without affecting the centroid frequency of the power spectrum. Bosentan administration induced a significant increase in tidal volume (V(T)), accompanied by a significant decrease in respiratory frequency, without any significant change in pulmonary ventilation, CO(2) arterial blood gas pressure or pH. Since the relationship between V(T) and inspiratory time remained substantially constant after bosentan administration, the changes in respiratory pattern appear to be the result of an upward shift in inspiratory off-switch threshold. Both inspiratory and expiratory times during occluded breathing were increased by block of ET receptors, suggesting also a central respiratory neuromodulator effect of ET. In conclusion the present results suggest that the block of ET receptors in spontaneously breathing pigs exerts a role on mechanical properties of the respiratory system as well as on peripheral and central mechanisms of breathing control. PMID- 11728167 TI - Influence of antioxidative vitamins A, C and E on lipid peroxidation in BOP induced pancreatic cancer in Syrian hamsters. AB - Persistent oxidative stress is thought to play an important role in carcinogenesis. Vitamins may influence oxygen radical metabolism and thus inhibit tumor growth. In the present trial the effects of Vitamins (Vit.) A, C and E on neoplastic growth and lipid peroxidation in pancreatic tissue were evaluated on chemically-induced pancreatic adenocarcinoma in the Syrian hamster. The incidence of pancreatic cancer was decreased by Vit. A (64.3%) and Vit. C (71.4%) as compared to the control group (100%, P<0.05). All vitamins increased the activity of superoxidedismutase (SOD) in pancreatic carcinomas. Accumulation of vitamins in tumor cells seems to be responsible for high levels of SOD and consecutive intracellular increase of hydrogen peroxide levels. Since this effect is selectively toxic for tumor cells it might be one of the mechanisms decreasing the incidence of pancreatic cancer in our trial. PMID- 11728168 TI - Identification of a tyrosine kinase-phosphorylated protein in arachidonic acid- and prostaglandin A(2)-treated cells in vitro. AB - The effects of 20 microg/ml exogenous arachidonic acid (AA) and prostaglandin A(2) (PGA(2)) were evaluated on total tyrosine kinase (TK) activity and tyrosine phosphorylation status in HeLa and MCF-7 cells. AA and PGA(2) increased TK activity in both HeLa and MCF-7 cells. Western blotting employing an anti phosphotyrosine antibody showed only one protein of approximately 55 kDa (approximately 55 kDa) to be phosphorylated in the MCF-7 cells, while a variety of proteins were phosphorylated in the HeLa cells, including the approximately 55 kDa protein. Amino acid analyses as well as Matrix Assisted Laser Desorption Ionization were conducted on this protein from different cell lines and it was shown to be similar. Comparison to p53 did not show similarities. The identity of this protein needs to be further characterized to help elucidate the signal transduction pathways of AA and PGA(2). PMID- 11728169 TI - Effects of nitric oxide donor and inhibitor on prostaglandin E2-like activity, malondialdehyde and reduced glutathione levels after skeletal muscle ischemia reperfusion. AB - Oxygen free radicals are implicated in the pathophysiology of ischemia reperfusion (I/R) injury in skeletal muscle. Nitric oxide (NO) and prostaglandin E2 (PGE2) are important regulators of the microcirculation in skeletal muscle. The effects of L-arginine, substrate for NO, and N(G)-nitro L-arginine methyl ester (L-NAME) on PGE2 synthesis, lipid peroxidation and reduced glutathione (GSH) levels was investigated in the rat gastrocnemius muscle after 3 h of reperfusion following 2 h of ischemia. Lipid peroxidation and GSH levels showed a non-significant changes in the I/R groups compared to the control group. According to these results, it can be assumed that skeletal muscle can resist 2 h of ischemia followed by 3 h of reperfusion-induced oxidative stress. PGE2-like activity in the gastrocnemius muscle increased in the L-NAME treated and I/R groups. L-arginine administration reversed the increase in PGE2-like activity of reperfused skeletal muscle. These findings support the conclusion that endothelium-derived PGE2 synthesis increases during reperfusion and suggest that PGE2 may have a protective role in the maintenance of endothelial function. PMID- 11728170 TI - Surfactant protein A inhibits the non-enzymatic lipid peroxidation of porcine lung surfactant. AB - The ability of surfactant protein A (SP-A) to inhibit the ascorbate-Fe(2+) induced lipid peroxidation of polyunsaturated fatty acids found in porcine lung surfactant (surfacen) was assessed by measuring the light emission - chemiluminescence during a 180-min incubation period at 37 degrees C. The light emission (chemiluminescence) was concentration dependent. Changes in the fatty acid composition of surfacen were observed when the lung surfactant was incubated in an ascorbate-Fe(2+) system. The main polyunsaturated fatty acids C18:2 n6 and C20:4 n6 found in the lung surfactant decreased considerably after a 180-min lipid peroxidation process. Native SP-A isolated from pig lungs inhibited oxidation of surfactant long chain polyunsaturated fatty acids, mainly arachidonic acid, in a dose-dependent fashion that was half-maximal (60% inhibition) at a concentration of 2.0 microg/ml and almost complete (73.6% inhibition) at 4.0 microg/ml, as indicated by inhibition of light emission and fatty acid composition analysis. At the highest concentration of lung SP-A used a very good correlation between the protection of the most polyunsaturated fatty acids and inhibition of light emission was observed. PMID- 11728171 TI - Relationship between thromboxane/prostacyclin ratio and diabetic vascular complications. AB - To elucidate the relationship between the thromboxane A2/prostacyclin (TXA2/PGI2) ratio and diabetic complications, the levels of 11-dehydro-thromboxane B2 and 2,3 dinor-6-keto-prostaglandin F1alpha, the urinary metabolites of thromboxane A2 and prostacyclin, were measured in diabetics by gas chromatography/selected ion monitoring. We compared the TXA2/PGI2 ratio in healthy volunteers and diabetics. The TXA2/PGI2 ratio of diabetics was significantly higher than that of healthy volunteers and we could reconfirm the hypercoagulable condition in diabetics. We also investigated the difference of TXA2/PGI2 levels in diabetics with retinopathy and neuropathy. The TXA2/PGI2 ratio of diabetics with retinopathy showed significantly higher level than without retinopathy. However, the TXA2/PGI2 ratio of diabetics with neuropathy was the same as without neuropathy. These results suggest that the TXA2/PGI2 ratio reflects the pathological conditions of diabetes, especially the change of vasculature. The monitoring and improvement of TXA2/PGI2 ratio could be useful for the prevention of diabetic vascular complications. PMID- 11728172 TI - Effect of corticosteroids and eicosapentaenoic acid/docosahexaenoic acid on pro oxidant and anti-oxidant status and metabolism of essential fatty acids in patients with glomerular disorders. AB - It is known that the concentrations of essential fatty acids and their metabolites including eicosanoids, free radicals and anti-oxidants are altered in glomerular disorders. Both corticosteroids and n-3 fatty acids--eicosapentaenoic acid and docosahexaenoic acid (EPA and DHA respectively)--are useful in the management of glomerular disorders. In the present study, the altered plasma concentrations of lipid peroxides, nitric oxide and the metabolites of essential fatty acids and anti-oxidants--superoxide dismutase, glutathione peroxidase and vitamin E--in the RBC membranes of patients with glomerular disorders (nephrotic syndrome) reverted to normalcy following corticosteroids or EPA/DHA administration. This suggests that the beneficial actions of corticosteroids and EPA/DHA in glomerular disorders can be attributed to their action on the pro oxidant and anti-oxidant concentrations and metabolism of essential fatty acids. PMID- 11728173 TI - Dietary gamma-linolenic acid dose-dependently modifies fatty acid composition and immune parameters in rats. AB - gamma-linolenic acid (GLA) has been reported to improve several inflammatory disorders through regulation of eicosanoid production. However, since GLA is a precursor of arachidonic acid, it may bring about increasing tissue arachidonic acid levels with subsequent pro-inflammatory events. To explore this possibility, we examined the effect of high-dose GLA acid on the fatty acid profile of immune cells, leukotriene B4 production by peritoneal exudate cells and immunoglobulin productivity of mesenteric lymph node lymphocytes of Sprague-Dawley rats. Male rats were fed 10% fat diets containing graded levels, 0, 20, 40 and 60% of GLA for 3 weeks. The results showed the distinction in activity of metabolizing GLA between immune cells and liver. Thus, in immune cells such as mesenteric lymph node and spleen lymphocytes and peritoneal exudate cells, more dihomo-gamma linolenic acid was found than in the liver. Leukotriene B4 production by peritoneal exudate cells was significantly suppressed when fed the highest level of GLA suggesting a lower risk of allergic reaction. Moreover, immunoglobulin productivity in mesenteric lymph node lymphocytes was promoted by dietary GLA. The present study indicates that a high dose of GLA may exert anti-inflammatory effects through suppression of leukotriene B4 release and strengthening of gut immune system, thus ameliorating allergic reaction. PMID- 11728174 TI - Effects of intravenous oxygen on prostacyclin and thromboxane formation in patients with peripheral occlusive arterial disease. AB - Oxygen infusion is used in complementary medicine for treatment of peripheral occlusive arterial disease. The mechanism of action is unknown. Thus, we determined the effects of oxygen infusion on prostacyclin, thromboxane and nitric oxide synthesis. Twelve patients with peripheral occlusive arterial disease received oxygen 40 ml/d intravenously for 3 weeks. Study parameters, analyzed by gas chromatography-mass spectrometry on day 1, 3, 10, 16, 21: 2,3-dinor-6-oxo PGF(1alpha), colour invisible 2,3-dinor-TXB2 and nitrate in one-hour-urine before and after oxygen infusion, reflecting prostacyclin, thromboxane and nitric oxide synthesis. Urinary 8-iso-PGF2alpha, indicating oxidative stress, was assessed in one patient. Urinary 2,3-dinor-6-oxo-PGF1alpha rose from baseline more than 4 fold after oxygen infusion. In contrast, urinary 2,3-dinor-TXB2 excretion remained unchanged. Oxygen infusion had no effect on urinary nitrate excretion. Urinary 8-iso-PGF(2alpha) was not influenced by oxygen infusion with and without diclofenac pretreatment. Our data demonstrate a shift of the prostacyclin/thromboxane ratio toward prostacyclin by oxygen infusion. Thus, a mechanism of action is provided and clinical trials with intravenous oxygen find a rational basis. PMID- 11728175 TI - Expression of adipogenesis markers in a murine stromal cell line treated with 15 deoxy Delta(12,14)-prostaglandin J2, interleukin-11, 9-cis retinoic acid and vitamin K2. AB - Recent studies have demonstrated that bone marrow stromal cells can undergo adipogenesis or osteoblastogenesis in vivo, and in vitro, and that peroxisome proliferator-activated receptor gamma (PPAR gamma) plays a central role in the control of adipocyte differentiation. In the present study, we treated a murine stromal cell line (TMS-14) with a cocktail of dexamethasone, insulin and glucose (DIG cocktail), which caused the cells to convert to fat-laden cells with adipocyte-like morphology. We also exposed TMS-14 cells to DIG cocktail followed by 15-deoxy Delta(12,14)-prostaglandin J2 (15d-PGJ2), a ligand of PPAR gamma, interleukin- 11 (IL-11), 9-cis retinoic acid (9-cis RA) and vitamin K2. 15d-PGJ2 enhanced DIG cocktail-induced adipogenesis, whereas IL-11, 9-cis RA and vitamin K2 each inhibited adipogenesis induced by DIG cocktail. The gene expressions of four adipogenesis markers, PPAR gamma 2, adipocyte P2 (aP2), adipocyte determination and differentiation factor 1 (ADD1), and fatty acid synthase (FAS) were enhanced by DIG cocktail and these expressions were more enhanced by 15d PGJ2, in contrast they were attenuated by 9-cis RA. IL-11 also attenuated the adipogenesis markers except ADD1. Western blotting showed that 15d-PGJ2 enhanced the levels of PPAR gamma, C/EBP alpha and RXR alpha proteins, while IL-11 and 9 cis RA decreased the level of PPAR gamma protein, but not C/EBP alpha protein and vitamin K2 decreased the level of C/EBP alpha protein. We also tested the effect of 15d-PGJ2 on osteoblastogenesis, using TMS-12 cells, another stromal cell clone from the same mouse, which differentiate into osteoblasts spontaneously. 15d-PGJ2 did not affect osteoblastogenesis, as detected by von Kossa staining and Cbfa-1 gene expression. These data indicate that 15d-PGJ2 enhances the expression of both PPAR gamma and C/EBP alpha and as a result it stimulates adipogenesis in murine bone marrow cells. PMID- 11728176 TI - PGl2 analogue mitigates the progression rate of renal dysfunction improving renal blood flow without glomerular hyperfiltration in patients with chronic renal insufficiency. AB - Renal blood flow decreases with the progression of chronic glomerulonephritis (CGN). This disease induces medullary ischemia and further renal dysfunction in patients with chronic renal insufficiency (CRI). Prostacyclin (PGI2), with its vasodilative action, increases renal blood flow (RBF) without increasing glomerular filtration rate (GFR). We therefore examined the possibility that PGI2 would mitigate the progression of renal dysfunction by increasing RBF in patients with CRI. Sixteen patients with progressive renal insufficiency (serum creatinine: 2.14+/-0.89 mg/dl) due to CGN were prospectively chosen for this study. The blood pressure was already under control using calcium channel blockers before and during this study in nine hypertensive patients. In the first 6 months the patients received a low-protein (0.6 g/kg/day) and low-salt (5.0 g/day) diet. In the next 6 months they received 60 microg/day of PGI2 analogue (Beraprost sodium) orally. GFR was determined by 24-hour creatinine clearance, and effective renal plasma flow (ERPF) was determined by 99mTc-MAG3 scintigraphy. Glomerular capillary pressure, the resistance ratio of afferent and efferent arterioles (R(A)/R(E)), and the other hemodynamic parameters from Gomez's estimation equation were determined at the start of this study, just before the administration of Beraprost and at the end of the study. The levels of GFR and ERPF were 34.6+/-12.4 and 140.6+/-52.1 ml/min at the start of this study respectively, and decreased to 28.0+/- 12.0 and 115.6+/-45.3 ml/min after the first 6 months without Beraprost. The levels of GFR and ERPF stayed at 28.1+/ 15.7 and 119.2+/-57.6 ml/min after the next 6 months with Beraprost in the same patients. R(A)/R(E) increased in the first 6 months from 7.9+/-3.6 to 10.8+/-8.6, but remained constant during 6 months of Beraprost administration, at 10.5+/-8.0. These data indicate that PGI2 analogue diminishes the vascular resistance of glomerular afferent and efferent arterioles regulating the decrease of renal blood flow without glomerular hyperfiltration, thus mitigating the progression rate of renal dysfunction. PMID- 11728177 TI - Stimulated release of arachidonic acid by agonists of the peroxisome proliferator activated receptor and retinoic acid receptors. AB - Release of arachidonic acid from rat liver cells is stimulated after a 6-hour incubation with 9-cis retinoic acid, all trans retinoic acid, the selective peroxisome proliferator-activated receptor-gamma synthetic thiazolidinedione, ciglitazone, the cyclopentenones, 15-deoxy-Delta(12,14) PGJ2 and PGA1 and the non steroidal anti-inflammatory drugs, celecoxib and indomethacin. The rates of the release stimulated by 15-deoxy-Delta(12,14) PGJ2 differ from those observed with celecoxib. Arachidonic acid release by9-cis retinoic acid in the presence of either ciglitazone or trans retinoic acid is synergistic. It is additive in the presence of celecoxib. Cycloheximide and actinomycin inhibit the release of arachidonic acid stimulated by 15-deoxy-Delta(12,14) PGJ2 but not by celecoxib. The findings indicate that agonists of the peroxisome proliferator-activated receptor-gamma and retinoic acid receptors stimulate the release of arachidonic acid. The mechanisms involved may differ in the cases of 15-deoxy-Delta(12,14) PGJ2 and celecoxib. PMID- 11728178 TI - Conformationally constrained analogues of diacylglycerol. 18. The incorporation of a hydroxamate moiety into diacylglycerol-lactones reduces lipophilicity and helps discriminate between sn-1 and sn-2 binding modes to protein kinase C (PK C). Implications for isozyme specificity. AB - An approach to reduce the log P in a series of diacylglycerol (DAG)-lactones known for their high binding affinity for protein kinase C (PK-C) is presented. Branched alkyl groups with reduced lipophilicity were selected and combined with the replacement of the ester or lactone oxygens by NH or NOH groups. Compound 6a with an isosteric N-hydroxyl amide arm represents the most potent and least lipophilic DAG analogue known to date. PMID- 11728179 TI - Discovery of small-molecule inhibitors of Bcl-2 through structure-based computer screening. AB - Bcl-2 belongs to a growing family of proteins which regulates programmed cell death (apoptosis). Overexpression of Bcl-2 has been observed in 70% of breast cancer, 30-60% of prostate cancer, 80% of B-cell lymphomas, 90% of colorectal adenocarcinomas, and many other forms of cancer. Thereby, Bcl-2 is an attractive new anti-cancer target. Herein, we describe the discovery of novel classes of small-molecule inhibitors targeted at the BH3 binding pocket in Bcl-2. The three dimensional (3D) structure of Bcl-2 has been modeled on the basis of a high resolution NMR solution structure of Bcl-X(L), which shares a high sequence homology with Bcl-2. A structure-based computer screening approach has been employed to search the National Cancer Institute 3D database of 206 876 organic compounds to identify potential Bcl-2 small-molecule inhibitors that bind to the BH3 binding site of Bcl-2. These potential Bcl-2 small-molecule inhibitors were first tested in an in vitro binding assay for their potency in inhibition of the binding of a Bak BH3 peptide to Bcl-2. Thirty-five potential inhibitors were tested in this binding assay, and seven of them were found to have a binding affinity (IC(50) value) from 1.6 to 14.0 microM. The anti-proliferative activity of these seven active compounds has been tested using a human myeloid leukemia cell line, HL-60, which expresses the highest level of Bcl-2 protein among all the cancer cell lines examined. Compound 6 was the most potent compound and had an IC(50) value of 4 microM in inhibition of cell growth using the 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Five other compounds had moderate activity in inhibition of cell growth. Compound 6 was further evaluated for its ability to induce apoptosis in cancer cells. It was found that 6 induces apoptosis in cancer cells with high Bcl-2 expression and its potency correlates with the Bcl-2 expression level in cancer cells. Furthermore, using NMR methods, we conclusively demonstrated that 6 binds to the BH3 binding site in Bcl-X(L). Our results showed that small-molecule inhibitors of Bcl-2 such as 6 modulate the biological function of Bcl-2, and induce apoptosis in cancer cells with high Bcl-2 expression, while they have little effect on cancer cells with low or undetectable levels of Bcl-2 expression. Therefore, compound 6 can be used as a valuable pharmacological tool to elucidate the function of Bcl-2 and also serves as a novel lead compound for further design and optimization. Our results suggest that the structure-based computer screening strategy employed in the study is effective for identifying novel, structurally diverse, nonpeptide small molecule inhibitors that target the BH3 binding site of Bcl-2. PMID- 11728180 TI - Free energy component analysis for drug design: a case study of HIV-1 protease inhibitor binding. AB - A theoretically rigorous and computationally tractable methodology for the prediction of the free energies of binding of protein-ligand complexes is presented. The method formulated involves developing molecular dynamics trajectories of the enzyme, the inhibitor, and the complex, followed by a free energy component analysis that conveys information on the physicochemical forces driving the protein-ligand complex formation and enables an elucidation of drug design principles for a given receptor from a thermodynamic perspective. The complexes of HIV-1 protease with two peptidomimetic inhibitors were taken as illustrative cases. Four-nanosecond-level all-atom molecular dynamics simulations using explicit solvent without any restraints were carried out on the protease inhibitor complexes and the free proteases, and the trajectories were analyzed via a thermodynamic cycle to calculate the binding free energies. The computed free energies were seen to be in good accord with the reported data. It was noted that the net van der Waals and hydrophobic contributions were favorable to binding while the net electrostatics, entropies, and adaptation expense were unfavorable in these protease-inhibitor complexes. The hydrogen bond between the CH2OH group of the inhibitor at the scissile position and the catalytic aspartate was found to be favorable to binding. Various implicit solvent models were also considered and their shortcomings discussed. In addition, some plausible modifications to the inhibitor residues were attempted, which led to better binding affinities. The generality of the method and the transferability of the protocol with essentially no changes to any other protein-ligand system are emphasized. PMID- 11728181 TI - Oxindole-based inhibitors of cyclin-dependent kinase 2 (CDK2): design, synthesis, enzymatic activities, and X-ray crystallographic analysis. AB - Two closely related classes of oxindole-based compounds, 1H-indole-2,3-dione 3 phenylhydrazones and 3-(anilinomethylene)-1,3-dihydro-2H-indol-2-ones, were shown to potently inhibit cyclin-dependent kinase 2 (CDK2). The initial lead compound was prepared as a homologue of the 3-benzylidene-1,3-dihydro-2H-indol-2-one class of kinase inhibitor. Crystallographic analysis of the lead compound bound to CDK2 provided the basis for analogue design. A semiautomated method of ligand docking was used to select compounds for synthesis, and a number of compounds with low nanomolar inhibitory activity versus CDK2 were identified. Enzyme binding determinants for several analogues were evaluated by X-ray crystallography. Compounds in this series inhibited CDK2 with a potency approximately 10-fold greater than that for CDK1. Members of this class of inhibitor cause an arrest of the cell cycle and have shown potential utility in the prevention of chemotherapy induced alopecia. PMID- 11728182 TI - [1,2,4]Triazino[4,3-a]benzimidazole acetic acid derivatives: a new class of selective aldose reductase inhibitors. AB - Acetic acid derivatives of [1,2,4]triazino[4,3-a]benzimidazole (TBI) were synthesized and tested in vitro and in vivo as a novel class of aldose reductase (ALR2) inhibitors. Compound 3, (10-benzyl[1,2,4]triazino[4,3-a]benzimidazol 3,4(10H)-dion-2-yl)acetic acid, displayed the highest inhibitory activity (IC(50) = 0.36 microM) and was found to be effective in preventing cataract development in severely galactosemic rats when administered as an eyedrop solution. All the compounds investigated were selective for ALR2, since none of them inhibited appreciably aldehyde reductase, sorbitol dehydrogenase, or glutathione reductase. The activity of 3 was lowered by inserting various substituents on the pendant phenyl ring, by shifting the acetic acid moiety from the 2 to the 3 position of the TBI nucleus, or by cleaving the TBI system to yield benzimidazolylidenehydrazines as open-chain analogues. A three-dimensional model of human ALR2 was built, taking into account the conformational changes induced by the binding of inhibitors such as zopolrestat, to simulate the docking of 3 into the enzyme active site. The theoretical binding mode of 3 was fully consistent with the structure-activity relationships in the TBI series and will guide the design of novel ALR2 inhibitors. PMID- 11728183 TI - Cheminformatic models to predict binding affinities to human serum albumin. AB - Models to predict binding affinities to human serum albumin (HSA) should be very useful in the pharmaceutical industry to speed up the design of new compounds, especially as far as pharmacokinetics is concerned. We have experimentally determined through high-performance affinity chromatography the binding affinities to HSA of 95 diverse drugs and druglike compounds. These data have allowed us the derivation of quantitative structure-activity relationship models to predict binding affinities to HSA of new compounds on the basis of their structure. Simple linear, one-variable models have been derived for specific families of compounds (r(2) > or = 0.80; q(2) > or = 0.62): beta-adrenergic antagonists, steroids, COX inhibitors, and tricyclic antidepressants. Also, global models have been derived to be applicable to the whole medicinal chemical space by using the full database of HSA binding constants described above. For this aim, a genetic algorithm has been used to exhaustively search and select for multivariate and nonlinear equations, starting from a large pool of molecular descriptors. The resulting models display good fits to the experimental data (r(2) > or = 0.78; LOF < or = 0.12). In addition, both internal (cross validation and randomization) and external validation tests have demonstrated that these models have good predictive power (q(2) > or = 0.73; PRESS/SSY < or = 0.23; r(2) > or = 0.82 for the external set). Statistical analysis of the equation populations indicates that hydrophobicity (as measured by the ClogP) is the most important variable determining the binding extent to HSA. In addition, structural factors (especially the topological (6)chi(ring) index and some Jurs descriptors) also frequently appear as descriptors in the best equations. Therefore, binding to HSA turns out to be determined by a combination of hydrophobic forces together with some modulating shape factors. This agrees with X-ray structures of HSA alone or bound to ligands, where the binding pockets of both sites I and II are composed mainly of hydrophobic residues. PMID- 11728184 TI - Systematic structure-based design and stereoselective synthesis of novel multisubstituted cyclopentane derivatives with potent antiinfluenza activity. AB - The design and synthesis of novel, orally active, potent, and selective inhibitors of influenza neuraminidase differing structurally from existing neuraminidase inhibitors are described. X-ray crystal structures of complexes of neuraminidase with known five- and six-membered ring inhibitors revealed that potent inhibition of the enzyme is determined by the relative positions of the interacting inhibitor substituents (carboxylate, glycerol, acetamido, hydroxyl) rather than by the absolute position of the central ring. This led us to design potential neuraminidase inhibitors in which the cyclopentane ring served as a scaffold for substituents (carboxylate, guanidino, acetamido, alkyl) that would interact with the four binding pockets of the neuraminidase active site at least as effectively as those of the established six-membered ring inhibitors such as DANA (2), zanamivir (3), and oseltamivir (4). A mixture of the isomers was prepared initially. Protein crystallography of inhibitor-enzyme complexes was used to screen mixtures of isomers in order to identify the most active stereoisomer. A synthetic route to the identified candidate 50 was developed, which featured (3 + 2) cycloaddition of 2-ethylbutyronitrile oxide to methyl (1S,4R)-4[(tert-butoxycarbonyl)amino]cyclopent-2-ene-1-carboxylate (43). Structures of the synthetic compounds were verified by NMR spectroscopy using nuclear Overhauser effect methodology. Two new neuraminidase inhibitors discovered in this work, 50 and 54, have IC(50) values vs neuraminidase from influenza A and B of <1 and <10 nM, respectively. These IC(50) values are comparable or superior to those for zanamivir and oseltamivir, agents recently approved by the FDA for treatment of influenza. The synthetic route used to prepare 50 and 54 was refined so that synthesis of pure active isomer 54, which has five chiral centers, required only seven steps from readily available intermediates. Further manipulation was required to prepare deoxy derivative 50. Because the activities of the two compounds are comparable and 54 [RWJ-270201 (BCX-1812)] is the easier to synthesize, it was selected for further clinical evaluation. PMID- 11728185 TI - Discovery of novel p-arylthio cinnamides as antagonists of leukocyte function associated antigen-1/intercellular adhesion molecule-1 interaction. 4. Structure activity relationship of substituents on the benzene ring of the cinnamide. AB - We have shown that p-arylthio cinnamides can inhibit the interaction of LFA-1 and ICAM-1, which is involved in cell adhesion and the inflammatory process. We now show that 2,3-disubstitution on the aryl portion of the cinnamide results in enhanced activity over mono substitution on the ring. The best 2,3-substituents were chlorine and trifluoromethyl groups. Compounds 39 and 40 which contain two CF3 groups have IC(50) values of 0.5 and 0.1 nM, respectively, in inhibiting JY8 cells expressing LFA-1 on their surface, from adhering to ICAM-1. The structure activity relationship (SAR) was examined using an NMR based model of the LFA-1 I domain/compound 31 complex. One of our compounds (38) was able to reduce cell migration in two different in vivo experiments. PMID- 11728186 TI - Synthesis and structure-activity relationships of N-(1-benzylpiperidin-4 yl)arylacetamide analogues as potent sigma1 receptor ligands. AB - A series of N-(1-benzylpiperidin-4-yl)arylacetamides were synthesized and evaluated for their binding properties for sigma1 and sigma2 receptors. In agreement with previously reported sigma1/sigma2 receptor binding data for N-(1 benzylpiperidin-4-yl)phenylacetamide, all of the N-(1-benzylpiperidin-4 yl)arylacetamide compounds reported below displayed higher affinity for sigma1 vs sigma2 receptors. Replacement of the phenyl ring of the phenylacetamide moiety with a thiophene, naphthyl, or indole aromatic ring had no significant effect on the sigma1 receptor affinity. Replacement of the phenyl ring with an imidazole or pyridyl aromatic ring resulted in a >60-fold loss in affinity for sigma1 receptors and no significant binding affinity for sigma2 receptors. Substitution on the aromatic ring of the benzyl group showed a similar or slightly decreased affinity for sigma1 receptors. Substitution on the aromatic rings of both the phenylacetamide moiety and the benzyl group with a halogen resulted in a similar affinity for sigma(1) receptors and a significantly increased affinity for sigma2 receptors. Comparative molecular field analysis revealed that electrostatic properties of the substituents in the phenylacetamide aromatic ring strongly influenced binding to sigma1 receptors. Compounds 1, 10, 18, 22, 37, and 40 showed the highest selectivity for sigma1 receptors with K(i) (sigma2) to K(i) (sigma(1)) ratios of 100, >92, >122, 77, 74, and 80, respectively. In agreement with previously reported results, the phenylacetamide analogues had no binding affinity for dopamine receptors (D2/D3). PMID- 11728187 TI - New antimitotic agents with activity in multi-drug-resistant cell lines and in vivo efficacy in murine tumor models. AB - During a screen for compounds that could inhibit cell proliferation, a series of new tubulin-binding compounds was identified with the discovery of oxadiazoline 1 (A-105972). This compound showed good cytotoxic activity against non-multi-drug resistant and multi-drug-resistant cancer cell lines, but its utility in vivo was limited by a short half-life. Medicinal chemistry efforts led to the discovery of indolyloxazoline 22g (A-259745), which maintained all of the in vitro activity seen with oxadiazoline 1, but also demonstrated a better pharmacokinetic profile, and dose-dependent in vivo activity. Over a 28 day study, indolyloxazoline 22g increased the life span of tumor-implanted mice by up to a factor of 3 upon oral dosing. This compound, and others of its structural class, may prove to be useful in the development of new chemotherapeutic agents to treat human cancers. PMID- 11728188 TI - trans-4-[4-(Methoxyphenyl)cyclohexyl]-1-arylpiperazines: a new class of potent and selective 5-HT(1A) receptor ligands as conformationally constrained analogues of 4-[3-(5-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)propyl]-1-arylpiperazines. AB - The present paper concerns the influence of conformational parameters on the recognition by rat 5-HT1A receptors of derivatives 4-[3-(5-methoxy-1,2,3,4 tetrahydronaphthalen-1-yl)propyl]-1-(2-pyridinyl)piperazine (1a) and 3-(5-methoxy 1,2,3,4-tetrahydronaphthalen-1-yl)-N-[2-(2-pyridyloxy)ethyl]propanamine (3b), two highly potent and selective 5-HT1A receptor ligands. Fifteen corresponding flexible and rigid analogues were prepared following several synthetic routes and were tested in binding assays with radioligands at 5-HT1A, D2, and alpha1 receptors from rat brain membranes. Among the new derivatives emerged trans-4-[4 (3-methoxyphenyl)cyclohexyl]-1-(2-pyridinyl)piperazine (trans-8a) and trans-N-[4 (3-methoxyphenyl)cyclohexyl]-2-(2-pyridyloxy)ethylamine (trans-8b). These compounds can be considered as conformationally constrained analogues of compounds 1a and 3a, respectively. In fact, compounds trans-8a and trans-8b showed a marked enhancement in 5-HT1A receptor affinity when compared to the corresponding cis isomers. Because compound trans-8a was a potent and selective 5 HT1A ligand (K(i), nM: 5-HT1A = 0.028, D2 = 2194, alpha1 = 767), it was chosen as a lead to prepare other analogues that were tested at 5-HT1A, D2, and alpha1 receptors from rat brain membranes, showing high affinity at the 5-HT1A and selectivity vs D2 and alpha1 receptors. Selected compounds were tested for their affinity at the human cloned 5-HT1A, alpha1a, alpha1b, alpha1d receptor subtypes. They were also submitted to the [35S]GTPgammaS binding assay stimulating the 5 HT1A receptor-mediated G-protein activation, therefore behaving as full or as partial agonists. Finally, the ability of iv administration of trans-8a to induce fore-paw treading in rats was evaluated in comparison with 8-OH-DPAT. Although the affinity (K(i)) and in vitro activity (pD'2) of trans-8a at the 5-HT1A receptor were higher than those of 8-OH-DPAT, the compound was less potent than the reference standard in inducing the symptom. PMID- 11728189 TI - Antimalarial alkoxylated and hydroxylated chalcones [corrected]: structure activity relationship analysis. AB - Chalcones with 2',3',4'-trimethoxy, 2',4'-dimethoxy, 4'-methoxy, 4'-ethoxy, 2',4' dihydroxy, and 4'-hydroxy groups on ring B were synthesized and evaluated in vitro against Plasmodium falciparum (K1) in a [3H] hypoxanthine uptake assay. The other ring A was quinoline, pyridine, naphthalene, or phenyl rings with electron donating or electron-withdrawing substituents of varying lipophilicities. Trimethoxy 6 and 27, dimethoxy 7, 8, 29, and methoxy 31 analogues had good in vitro activities (IC(50) < 5 microM). 3-Quinolinyl ring A derivatives were well represented among the active compounds. Hydroxylated chalcones were less active than the corresponding alkoxylated analogues. When evaluated in vivo, 8 and 208 were comparable to chloroquine in extending the lifespan of infected mice. Multivariate data analysis showed that in vitro activity was mainly determined by the properties of ring B. Quantitative structure-activity relationship models with satisfactory predictive ability were obtained for various B ring chalcones using projections to latent structures. A model with good predictability was proposed for 19 active chalcones. Size and hydrophobicity were identified as critical parameters. PMID- 11728190 TI - Novel tropane-based irreversible ligands for the dopamine transporter. AB - 3alpha-(diphenylmethoxy)tropane (benztropine) and its analogues are tropane ring containing dopamine uptake inhibitors that display binding and behavioral profiles that are distinct from cocaine. We previously prepared a benztropine based photoaffinity label [125I]-(N-[4-(4'-azido-3'-iodophenyl)butyl]-3alpha [bis(4'-fluorophenyl)methoxy]tropane, [125I]1, that covalently attached to the 1 2 transmembrane spanning region of the dopamine transporter (DAT). This was in contrast to the 4-7 transmembrane spanning region labeled by a cocaine-based photoaffinity label, [125I] 2 (RTI 82). To characterize further these different binding domains, photoaffinity ligands that had the 4'-azido-3'-iodophenyl substituent extended from the same position on the tropane ring were desirable. Thus, identification of the optimal alkyl linker between this substituent and the tropane nitrogen in the benztropine series was investigated to ultimately prepare the identical N-substituted analogue of 2. In this pursuit, the N-[4-(4'-azido-3' iodophenyl)propyl] analogue of 3alpha-[bis(4'-fluorophenyl)methoxy]tropane (9a) was synthesized as well as two isothiocyanate analogues that do not require photoactivation (10a,b) for irreversible binding. The synthesis of these target compounds was achieved using a modification of the strategy developed for 1. Evaluation of these compounds for displacing [3H]WIN 35 428 binding at DAT in rat caudate putamen revealed that the 4'-azido-3'-iodophenylbutyl substituent, found in 1, provided optimal binding affinity and was chosen to replace the N-CH3 group on 2. Both the 4'-azido-3'-iodophenyl- and the 4'-isothiocyanatophenylbutyl analogues of 2 (25 and 26, respectively) were synthesized. Both products bound to DAT with comparable potency (IC(50) = 30 nM) to RTI 82 (2). In addition, compound 26 demonstrated wash-resistant displacement of [3H]WIN 35 428 in HEK 293 cells stably transfected with hDAT. These ligands will provide important tools for further characterizing the binding domains for tropane-based dopamine uptake inhibitors at the DAT. PMID- 11728191 TI - O-Phosphonatomethylcholine, its analogues, alkyl esters, and their biological activity. AB - O-Phosphonatomethylcholine, an isopolar phosphocholine analogue with a phosphonomethyl ether group replacing a phosphomonoester residue, was prepared by reaction of diisopropyl 2-chloroethoxymethylphosphonate with dimethylamine followed by quaternization of the thus-obtained diisopropyl 2 dimethylaminoethoxymethylphosphonate with iodomethane; the ester groups in the quaternary intermediate were cleaved with bromotrimethylsilane. Replacement of dimethylamine in the reaction sequence by morpholine and/or pyrrolidine gave the N-methylmorpholinium or N-methylpyrrolidinium analogues of O phosphonatomethylcholine. Reaction of O-phosphonomethylcholine monotetrabutylammonium salt with 1-bromoalkanes in acetonitrile afforded a series of the corresponding monoalkyl (C10-C16) esters. None of these compounds except for the hexadecyl ester exhibited any appreciable cytostatic activity against DU 145, H460, HT-29, or MES-SA cell lines in vitro (evaluated by 3H-Thd incorporation assay). The hexadecyl ester exhibited modest in vitro cytotoxic activity comparable to that of the anticancer drug miltefosine (hexadecyl O phosphocholine). In vivo evaluation of hexadecyl O-phosphonomethylcholine [transplanted SD lymphoma in inbred SD/cub rats, 10 mg kg(-1) day(-1) intratumoral injection for 10 days] resulted in a 40% decrease in lymphoma mass. PMID- 11728192 TI - 3-(Arylacetylamino)-N-methylbenzamides: a novel class of selective anti Helicobacter pylori agents. AB - After chemical modification preceded by the random screening of our chemical library, a novel class of selective anti-Helicobacter pylori agents was generated. Consequently, the 3-(arylacetylamino)-N-methylbenzamides, which were quite easy to prepare, showed potent inhibitory activity against Helicobacter pylori but exhibited no inhibitory activity against other sorts of bacteria and fungi, e.g., Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Bacteroides fragilis, and Candida albicans. These compounds showed potent anti-H. pylori activity under acidic conditions, whereas amoxicillin and clarithromycin decreased activity. The 3-(3-arylpropionylamino)-N methylbenzamides, 3-(aryloxyacetylamino)-N-methylbenzamides, and (3 methylcarbamoylphenyl)carbamic acid 1-arylmethyl esters also exhibited potent anti-H. pylori activity. Finally, we selected 7n (BAS-118) as a candidate compound for further evaluation. PMID- 11728193 TI - Synthesis and biological studies of novel nucleoside phosphoramidate prodrugs. AB - A novel approach to the intracellular delivery of nucleotides using phosphoramidate-based prodrugs is described. Specifically, we have developed phosphoramidate prodrugs of the anticancer nucleotide 5-fluoro-2'-deoxyuridine 5'monophosphate (FdUMP). These phosphoramidate prodrugs contain an ester group that undergoes intracellular activation liberating phosphoramidate anion, which undergoes spontaneous cyclization and P-N bond cleavage to yield the nucleoside monophosphate quantitatively. In vitro evaluation of 5-fluoro-2'-deoxyuridine phosphoramidate prodrugs 2a and 3b against L1210 mouse leukemia cells show potent inhibition of cell growth (IC(50) 0.5-3 nM). Cell-based thymidylate synthase inhibition studies show that, in contrast to FUdR, the nitrofuran compound 2a is of comparable potency in wild type vs thymidine kinase deficient LM cells. This result indicates that the activation of this novel prodrug occurs via the proposed mechanism of intracellular delivery. However, naphthoquinone 3b has an IC(50) value for thymidylate synthase inhibition that is comparable to FUdR in thymidine kinase deficient cells. Further studies revealed that 3b rapidly decomposes to the nucleotide in cell culture medium, suggesting that the naphthoquinone analogue is not sufficiently stable to function as a nucleotide prodrug. PMID- 11728194 TI - Nonsteroidal selective glucocorticoid modulators: the effect of C-5 alkyl substitution on the transcriptional activation/repression profile of 2,5-dihydro 10-methoxy-2,2,4-trimethyl-1H-[1]benzopyrano[3,4-f]quinolines. AB - The preparation and characterization of a series of selective glucocorticoid receptor modulators are described. The preliminary structure-activity relationship of nonaromatic C-5 substitution on the tetracyclic quinoline core showed a preference for small lipophilic side chains. Proper substitution at this position maintained the transcriptional repression of proinflammatory transcription factors while diminishing the transcriptional activation activity of the ligand/glucocorticoid receptor complex. The optimal compounds described in this study were the allyl analogue 18 and cyclopentyl analogue 32. These candidates showed slightly less potent, highly efficacious E-selectin repression with significantly reduced levels of glucocorticoid response element activation in reporter gene assays vs prednisolone. Allyl analogue 18 was evaluated in vivo. An oral dose of 18 showed an ED(50) = 1.7 mg/kg as compared to 1.2 mg/kg for prednisolone in the Sephadex-induced pulmonary eosinophilia model and an ED(50) = 15 mg/kg vs 4 mg/kg for prednisolone in the carrageenan-induced paw edema model. PMID- 11728195 TI - Design, synthesis, and biological activity of a novel non-cisplatin-type platinum acridine pharmacophore. AB - Platinum-acridine conjugates were prepared from [PtCl2(ethane-1,2-diamine)] and the novel acridinylthioureas MeHNC(S)NMeAcr (6) and MeHNC(S)NMe(CH2CH2)NHAcr (15) by replacing one chloro leaving group in the cisplatin analogue with thiourea sulfur. In HL-60 leukemia cells, IC(50) values for 7 (Pt-tethered 6) and 16 (Pt tethered 15) were 75 and 0.13 microM, respectively. In the ovarian cell lines 2008 and C13, 16 was active at micromolar concentrations and showed only partial cross-resistance with clinical cisplatin. Possible structure-activity relationships are discussed. PMID- 11728196 TI - Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications. AB - OBJECTIVE: At 380 nm excitation, cervical tissue fluorescence spectra demonstrate characteristic changes with both patient age and the presence of dysplasia. A Monte Carlo model was developed in order to quantitatively examine how intrinsic NADH and collagen fluorescence, in combination with tissue scattering and absorption properties, yield measured tissue spectra. METHODS: Excitation emission matrices were measured for live cervical cells and collagen gel phantoms. Fluorescence microscopy of fresh tissue sections was performed to obtain the location and density of fluorophores as a function of patient age and the presence of dysplasia. A Monte Carlo model was developed which incorporated measurements of fluorophore line shapes and spatial distributions. RESULTS: Modeled spectra were consistent with clinical measurements and indicate that an increase in NADH fluorescence and decrease in collagen fluorescence create clinically observed differences between normal and dysplastic tissue spectra. Model predictions were most sensitive to patient age and epithelial thickness. CONCLUSIONS: Monte Carlo techniques provide an important means to investigate the combined contributions of multiple fluorophores to measured emission spectra. The approach will prove increasingly valuable as a more sophisticated understanding of in vivo optical properties is developed. PMID- 11728197 TI - Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions. AB - Immature and dysplastic cervical squamous epithelium whitens after the application of acetic acid during a colposcopic examination. The whitening process occurs visually over several minutes and subjectively discriminates between dysplastic and normal tissue. In this work, examples of the acetowhitening process are detailed in three ways: the color-imaged colposcopic appearance of the acetowhitening of high-grade cervical intraepithelial neoplasia (CIN 2/3), the kinetics of these reflectance patterns transformed to reduce noise in the signal, and a self-normalized green to red ratio measurement of the kinetics of these reflectance patterns. A total of six patients with biopsy confirmed CIN 2/3 were examined to obtain a set of timed images tracking the acetowhitening and the whitening-decay process over the course of 5-10 min. Regions of normal mature squamous epithelium within the same patients were also followed as an internal control. We determined that the temporal change over a 10 min time period in the ratio of green to red light intensities, taken from the respective color channels of the CCD, provides a reliable measure to clearly distinguish CIN 2/3 from normal cervical epithelium. This imaging and data normalization procedure may be applied to cervical lesions of different grades, to determine if a quantitative estimate provides predictive value during the colposcopic diagnosis. PMID- 11728198 TI - Optical characterization of melanin. AB - The optical properties of melanin have been characterized for a number of laser wavelengths in the visible region. The index of refraction of melanin is measured by the conventional method of minimum deviation using a hollow quartz prism at these wavelengths. The inverse adding doubling method based on the diffusion approximation and radiative transport theory have been employed to determine the absorption, scattering, and scattering anisotropy coefficients of melanin from the measurements of diffuse transmission, diffuse reflection and collimated transmission using double integrating spheres. The results obtained by the use of inverse adding doubling method have been compared to the Monte Carlo simulation technique. PMID- 11728199 TI - Real-time pachymetry during photorefractive keratectomy using optical low coherence reflectometry. AB - Optical low-coherence reflectometry (OLCR) was used as a noncontact method to measure the central corneal thickness of three patients intraoperatively during photorefractive keratectomy. Continuous on-line measurements were performed on the intact cornea immediately before the beginning of surgery, after the removal of the corneal epithelium, during laser tissue photoablation, and for 3 min after the ablation process. Corneal thinning due to evaporation was studied on a separate patient with the OLCR instrument, and it was found to be -0.14 microm/s during the first 5 min after epithelium removal. This baseline corneal thinning rate was used as a fit parameter to calculate actual from measured ablation depths. The measurements showed a maximum difference of +/-10 microm between planned ablations (34-92 microm) and measured ablation depths. PMID- 11728200 TI - Processing algorithms for tracking speckle shifts in optical elastography of biological tissues. AB - Parametric and nonparametric data processing schemes for analyzing translating laser speckle data used to investigate the mechanical behavior of biological tissues are examined. Cross-correlation, minimum mean square estimator, maximum likelihood, and maximum entropy approaches are discussed and compared on speckle data derived from cortical bone samples undergoing dynamic loading. While it was not the purpose of this paper to demonstrate that one processing technique is superior to another, maximum likelihood and maximum entropy approaches are shown to be particularly useful when the observed speckle motion is small. PMID- 11728201 TI - Surgical adhesives for laser-assisted wound closure. AB - Solid protein solder-doped polymer membranes were developed for laser-assisted tissue repair. Biodegradable polymer membranes of controlled porosity were fabricated with poly(L-lactic-co-glycolic acid) (PLGA), poly(ethylene glycol) (PEG), and salt particles, using a solvent-casting and particulate-leaching technique. The membranes provided a porous scaffold that readily absorbed the traditional protein solder composed of serum albumin, indocyanine green dye, and de-ionized water. In vitro investigations were conducted to assess the influence of various processing parameters on the strength of tissue repairs formed using the new membranes. These parameters included PLGA copolymer and PLGA/PEG blend ratios, membrane pore size, initial albumin weight fraction, and laser irradiance used to denature the solder. Altering the PLGA copolymer ratio had little effect on repair strength, however such variations are known to influence the degradation rate of the membranes. The repair strength increased with increased membrane pore size and bovine serum albumin concentration. The addition of PEG during the membrane casting stage increased the flexibility of the membranes but not necessarily the repair strength. Typically, the repair strength increased with increasing irradiance from 12 to 18 W/cm(2). The new solder-doped polymer membranes provided all of the benefits associated with solid protein solders including high repair strength and improved edge coaptation. In addition, the flexible, moldable nature of the new membranes offers the capability of tailoring the membranes to a wide range of clinically relevant geometries. PMID- 11728203 TI - Depth dependence of the analytical expression for the width of the point spread function (spatial resolution) in time-resolved transillumination. AB - Simple analytical expressions for the point spread function (PSF) at different depths can save computation time and improve the performance of inverse algorithms for optical imaging. In particular, application of such formulas simplifies quantification of the optical characteristics of tissue abnormalities inside highly scattering media. Earlier it was shown within the random walk theory framework that the PSF for time-resolved transillumination imaging through a highly scattering slab is well represented by a Gaussian. We have experimentally validated a simple formula of the random walk model for the effective width of this Gaussian, as a function of time delay, at different depths of the target. Presented analysis of published experimental data, concerning effective width of the PSF, for a slab of considerably smaller thickness also demonstrates good agreement between the data and predictions of our model. This PSF width determines spatial resolution of the time-resolved imaging and is widely discussed in the literature. PMID- 11728202 TI - In vivo imaging of light-emitting probes. AB - In vivo imaging of cells tagged with light-emitting probes, such as firefly luciferase or fluorescent proteins, is a powerful technology that enables a wide range of biological studies in small research animals. Reporters with emission in the red to infrared (>600 nm) are preferred due to the low absorption in tissue at these wavelengths. Modeling of photon diffusion through tissue indicates that bioluminescent cell counts as low as a few hundred can be detected subcutaneously, while approximately 10(6) cells are required to detect signals at approximately 2 cm depth in tissue. Signal-to-noise estimates show that cooled back-thinned integrating charge coupled devices (CCDs) are preferred to image intensified CCDs for this application, mainly due to their high quantum efficiency (approximately 85%) at wavelengths >600 nm where tissue absorption is low. Instrumentation for in vivo imaging developed at Xenogen is described and several examples of images of mice with bioluminescent cells are presented. PMID- 11728204 TI - In vitro optical characterization and discrimination of female breast tissue during near infrared femtosecond laser pulses propagation. AB - Ultrashort infrared laser pulses were transmitted through excised female breast tissue. The resulted signal was recorded by a streak camera with a time resolution of the order of a few ps. Experimental data of the temporal spread of the ultrashort pulse during the transmission through the tissue have been analyzed using the Patterson analytical expression derived from the diffusion theory. This resulted in the calculation of the absorption and reduced scattering coefficients, which are related to the optical characteristics of each type of tissue. The goal of the study was to use the theoretical values of the coefficients to discriminate different kinds of tissue. PMID- 11728205 TI - Novel surface and multicolor charge coupled device-based fluorescent imaging system for DNA microarrays. AB - We report a novel support, concomitant attachment chemistry, and a fluorescent imaging system for DNA microarrays. The support consists of soda lime glass coated with a layer of chromium, which eliminates any autofluorescence from the underlying glass substrate and reduces nonspecific probe binding. Attachment of DNA fragments exceeding 300 nucleotides in length is achieved without chemical modifications of either the chrome surface or the DNA itself. The charge coupled device (CCD)-based imaging system employs a 175 W xenon arc lamp as the light source, allowing the use of many different fluorophors. A 14 mm x 9 mm sample area is imaged with a single exposure, which takes between 5 and 20 s for each color plane in typical genomic comparative genomic hybridization type assays. The spatial resolution is limited only by the pixel size of the CCD chip (9 microm). The oblique illumination geometry combined with effective background reduction afforded by the chromium surface enables the system to achieve a detection limit of <5 x 10(7) fluorophors/cm(2) with 10 s integration. In a model system with arrayed lambda DNA targets a dose response was observed over four orders of magnitude in response to hybridizations with increasing amounts of the fluorescent labeled lambda probe. PMID- 11728206 TI - Evaluation of blood oxygen saturation in vivo from diffuse reflectance spectra. AB - A simple method to evaluate the hemoglobin oxygen saturation and relative hemoglobin concentration in a tissue from diffuse reflectance spectra in the visible wavelength range is put forward in this paper. It was assumed that while oxygenated and deoxygenated hemoglobin contributions to light attenuation are strongly variable functions of wavelength, all other contributions to the attenuation including scattering are smooth wavelength functions and can be approximated by Taylor series expansion. Based on this assumption, a simple, robust algorithm suitable for real time monitoring of the hemoglobin oxygen saturation in the tissue has been derived. This algorithm can be used with different fiber probe configurations for delivering and collecting light passed through the tissue. An experimental technique using this algorithm has been developed for in vivo monitoring during artery occlusion and in vitro monitoring of blood samples. The experimental results obtained are presented in the paper. PMID- 11728207 TI - Experimental verification of the effect of refractive index mismatch on the light fluence in a turbid medium. AB - Diffusion theory is often used to model the transport of light within tissue. It can be used to calculate the light fluence rate in tissue, for example, during photodynamic therapy, or to measure the absorption and scattering properties of tissue. For both of these applications, the influence of the interface between the tissue and the exterior medium on the fluence rate inside the tissue must be known in order to make accurate calculations. We present an experimental investigation of the effect of the refractive index mismatch at the tissue interface on the internal light fluence rate and on the spatially resolved diffuse reflectance as the boundary conditions of the tissue/external medium are changed. The effects of changing the relative refractive index at the boundary are compared to predictions of diffusion theory. The effect of the refractive index mismatch is predicted correctly by diffusion theory. PMID- 11728208 TI - In vivo burn depth determination by high-speed fiber-based polarization sensitive optical coherence tomography. AB - We report the first application of high-speed fiber-based polarization sensitive optical coherence tomography (PS-OCT) to image burned tissue in vivo. Thermal injury denatures collagen in skin and PS-OCT can measure the reduction in collagen birefringence using depth resolved changes in the polarization state of light propagated in, and reflected from, the tissue. Stokes vectors were calculated for each point in a scan and birefringence relative to incident polarization determined using four incident polarization states. Using a high speed fiber-based PS-OCT system on rat skin burned for varying periods of time, a correlation between birefringence and actual burn depth determined by histological analysis was established. In conclusion, PS-OCT has potential use for noninvasive assessment of burn depth. PMID- 11728210 TI - The impact of genomics on the biotechnology industry. PMID- 11728211 TI - Development of lipophilic cations as therapies for disorders due to mitochondrial dysfunction. AB - Mitochondrial dysfunction causes or exacerbates a number of diseases. These include genetic disorders such as Friedreich's ataxia where the primary lesion is a defect in a nuclear gene and those diseases caused by mutations to mitochondrial DNA. Mitochondrial damage also contributes to neurodegenerative diseases, diabetes and ischaemia-reperfusion injury. Drug therapies to prevent or alleviate mitochondrial dysfunction use redox active compounds, anti-oxidants or mitochondrial co-factors, however, their effectiveness is limited. A promising approach to increase the selectivity and potency of these compounds is to modify them so that they concentrate within mitochondria. This can be done by incorporating a lipophilic cation which causes the molecules to concentrate several hundred-fold in mitochondria, driven by the membrane potential across the inner membrane. As lipophilic cations cross biological membranes easily, they can be delivered to mitochondria of the heart, brain and skeletal muscle, the organs most affected by mitochondrial damage. Mitochondria-targeted lipophilic cations may lead to improved therapies for diseases involving mitochondrial dysfunction. PMID- 11728212 TI - Bacterial ghosts as carrier and targeting systems. AB - Bacterial ghosts are empty cell envelopes originating from Gram-negative bacteria. They have a natural outer surface make-up which provides them with the original targeting functions of the bacteria they are derived from and are thus able to bind to and/or are taken up by specific cells or tissues of animal, human or plant origin. The extended bacterial ghost system represents a platform technology for creating new qualities in non-living carriers which can be used for the specific targeting of drugs, DNA or other compounds to overcome toxic or non-desired obstacles. Freeze dried bacterial ghosts are stable without the requirement of a cold chain and can be effectively administered orally and aerogenically as drug carriers. The new system is an alternative to liposomes and may have an advantage due to its higher specificity for targeting specific tissues, its easy method of production and its versatility in entrapping and packaging various compounds in different compartments of the carriers. PMID- 11728213 TI - Vector-mediated drug delivery to the brain. AB - As a consequence of the growing ageing population, many neurodegenerative diseases, cancer and infections of the brain will become more prevalent. Despite major advances in neuroscience, many potential therapeutic agents are denied access to the central nervous system (CNS) because of the existence of the blood brain barrier (BBB). This barrier is formed by the endothelial cells of the brain capillaries and its primary characteristic is the impermeability of the capillary wall due to the presence of complex tight junctions and a low endocytic activity. The BBB behaves as a continuous lipid bilayer and prevents the passage of polar and lipid-insoluble substances. The BBB is, therefore, the major obstacle to drugs that are potentially useful for combating diseases affecting the CNS. Extensive efforts have been made to develop CNS drug delivery strategies in order to enhance delivery of therapeutic molecules across the BBB. The current challenge is to develop drug-delivery strategies that will allow the passage of therapeutic drugs through the BBB in a safe and effective manner. This review focuses specifically on the strategies developed to enhance drug delivery across the BBB with an emphasis on the vector-mediated strategy. PMID- 11728214 TI - Successful vaccination for Lyme disease: a novel mechanism? AB - Borrelia burgdorferi sensu lato is the aetiologic agent of Lyme disease, which is a multi-system disorder resulting from the transmission of organisms from an infected tick. According to the US Centers for Disease Control, the incidence of Lyme disease in the US has increased 25-fold since national surveillance began and the geographical spread of Lyme disease causing spirochetes would indicate that the annual number of cases will continue to rise. Humoral immunity has been shown to play a role in protection and this has spurred efforts towards developing a Lyme disease vaccine. A number of protective immunogens have been characterised to date, but due to the heterogeneity of Lyme disease Borreliae, no single molecule has proven to be completely effective as a vaccinogen. This review will describe the immunogens that have been used to protect against B. burgdorferi infection, with a focus on the inherent challenges involved with providing successful immunity to B. burgdorferi. In addition, the promising aspects and the limitations of each protective immunogen will be discussed. PMID- 11728215 TI - Vaccination against Helicobacter pylori--an old companion of man. AB - Helicobacter pylori infection induces an important systemic and mucosal antibody response and a predominant Th1 cellular response. These immune responses, although powerful, fail to eliminate the infection. Studies in animals have shown that prophylactic and therapeutic immunisations are efficacious, although complete protective immunity has usually not been achieved. Initial human trials with recombinant urease showed that a mucosal immune response can be obtained following immunisations, with a decrease in bacterial density, but successful immunisation is still awaited. Progress is being made in several areas of vaccine design. A human vaccine against H. pylori would be favourable in terms of health benefits and costs in developed and developing countries. PMID- 11728216 TI - Development of immunotherapeutic strategies for HIV-1. AB - In the majority of untreated patients, HIV-1 infection presents as a progressive disease of the immune system. Recent studies indicate that immune responses can be induced in HIV-1 infected individuals, leading to some immune control of virus replication. Such immune responses are also observed in small numbers of untreated HIV-1 infected long-term non-progressor (LTNP) patients, as well as in other viral infections (including those with human herpes viruses). Emerging novel technologies, animal studies and detailed immunological studies have proven invaluable in defining the immune responses that are associated with a favourable clinical outcome. Central effector and regulatory cells are HIV-1-specific CD8+ cytotoxic T-lymphocytes (CTL) and CD4+ helper T-lymphocytes respectively. Fully functional antigen-presenting cells (APC) are also essential in all stages of HIV 1 infection and possibly some (but not all) antibody responses contribute to beneficial immunity. The availability of combination anti-retroviral drug therapy, which successfully controls viraemia, has enabled a beneficial outcome in many HIV-1 infected individuals. Since no chronically HIV-1 infected patient has been shown to eradicate virus, novel approaches utilising therapeutic immunisation and various cytokines to manipulate immune responses and to induce and steer immunity towards a desired phenotype are required. There is a clear rationale for immunotherapeutic intervention in chronic progressive HIV-1 infection, which forms the foundation for novel approaches aimed at inducing and maintaining immune control. Here we review the immunopathogenesis of HIV-1 infection and discuss the promises of therapeutic immunisation and immunotherapy in general and their potential in the treatment of chronic HIV-1 disease. PMID- 11728217 TI - Anticytokine therapy for osteoarthritis. AB - Osteoarthritis (OA) is a joint disease that involves degeneration of articular cartilage, weakening of the subchondral bone and limited intra-articular inflammation manifested by synovitis. Since the pathogenesis of OA involves multiple aetiologies, including mechanical, biochemical and genetic factors, it has been difficult to identify unique targets for therapy. Current pharmacological interventions focus primarily on improving symptoms. The rationale for the use of anticytokine therapy in OA is based on evidence from studies in vitro and in vivo that interleukin-1 (IL-1) and tumour necrosis factor (TNF)-alpha are the predominant pro-inflammatory and catabolic cytokines involved in the initiation and progression of articular cartilage destruction. Since the increased levels of catabolic enzymes, prostaglandins, nitric oxide (NO) and other markers in OA fluids and tissues appear to be related to elevated levels of IL-1 and TNF-alpha, therapies that interfere with the expression or actions of these cytokines are most promising. Other cytokines that are anti-inflammatory and are often detected, paradoxically, in OA tissues are also potential therapeutic agents for counteracting the cartilage destruction in OA. Identification of methods for early diagnosis is of key importance, since therapeutic interventions aimed at blocking or reversing structural damage will be more effective when there is the possibility of preserving normal homeostasis. At later stages, cartilage tissue engineering with or without gene therapy will also require anticytokine therapy to block damage to newly repaired cartilage. This review will focus on experimental approaches currently under study that may lead to elucidation of effective strategies for therapy in OA, with special emphasis on anticytokine therapy. PMID- 11728218 TI - Haemoglobin-based erythrocyte transfusion substitutes. AB - Concerns about the infectious and immunosuppressive risks of allogeneic blood products persist, and the increased disproportion of blood donation and consumption has reinforced the search for alternative erythrocyte transfusion strategies in recent years. With the absence of problems such as nephro-toxicity, increased colloid osmotic pressure and sudden renal clearance, modern haemoglobin based oxygen carriers (HBOC) have shown their effectiveness and tolerability in numerous animal and several clinical studies. HBOC can be infused without prior cross-matching and are now available as stable formulations with long shelf-life. Most clinical studies have been performed with human cross-linked haemoglobin (DCLHb) but all trials were stopped two years ago because of an increased mortality in two clinical trials in patients who received DCLHb after stroke and multiple injury shock. However, experimental trials in animals are in progress with DCLHb and recombinant human haemoglobin. In contrast, Phase III studies with polymerised bovine haemoglobin (HBOC-201) are finished or currently under evaluation showing that infusion of HBOC-201 can avoid or reduce allogeneic blood transfusion needs in specific peri-operative settings. As a consequence, HBOC 2001 was actually approved for treatment of peri-operative anaemia in elective adult surgical patients in South Africa. Other human or bovine haemoglobin solutions are currently being investigated in different clinical studies in cardiac surgery patients, sepsis and tumour patients. More recent investigations have shown that HBOC are not only simple erythrocyte transfusion substitutes but highly effective oxygen donators in terms of tissue oxygenation. HBOC open the door for a new therapeutic strategy: plasmatic oxygen delivery with physiological concentrations of inspired oxygen. In specific situations (e.g., ischaemia or arterial stenosis) HBOC have advantages over red blood cells because they can reach post-stenotic or poorly perfused tissues with the plasma stream, where erythrocytes are not able to pass. In addition to significant plasmatic oxygen transport, HBOC also enhance tissue oxygenation because of the facilitated oxygen release by HBOC and from remaining erythrocytes. Further studies will show, if the outcome of patients with impaired perfusion (e.g., stroke or myocardial infarction) can be improved by prophylactic or therapeutic application of HBOC. Whenever these formulations are globally launched, they will find differential indications as potent oxygen-delivering drugs in addition to the globally recognised goal of red cell substitutes in cases of bleeding. PMID- 11728219 TI - Recombinant antibodies for cancer diagnosis and therapy. AB - Recombinant antibodies now represent over 30% of biopharmaceuticals in clinical trials, highlighted by the recent approvals for cancer immunotherapy from the FDA which has awoken the biotechnology industry. Sales of these antibodies are increasing very rapidly to a predicted US$ 3 billion per annum worldwide by 2002. Since the development of new therapeutic reagent into commercial product takes 10 years, the recent FDA-approved antibodies are based on early antibody designs which are now considered primitive. Emerging technologies have created a vast range of novel, recombinant, antibody-based reagents which specifically target clinical biomarkers of disease. In the past year, radiolabelling of antibodies has increased their potential for cancer imaging and targeting. Recombinant antibodies have also been reduced in size and rebuilt into multivalent molecules for higher affinity. In addition, antibodies have been fused with many molecules including toxins, enzymes and viruses for prodrug therapy, cancer treatment and gene delivery. Recombinant antibody technology has enabled clever manipulations in the construction of complex antibody library repertoires for the selection of high-affinity reagents against refractory targets. Although phage display remains the most extensively used method, this year high affinity reagents have been isolated using alternative display and selection systems such as ribosome display and yeast display confirming the emergence of new display methods. Furthermore, innovative affinity maturation strategies have been developed to obtain high affinity reagents. This review focuses on developments in the last 12 months and describes the latest developments in the design, production and clinical use of recombinant antibodies for cancer diagnosis and therapy. PMID- 11728220 TI - Gene therapy for lysosomal storage disorders. AB - The lysosomal storage disorders (LSD) are monogenic inborn errors of metabolism with heterogeneous pathophysiology and clinical manifestations. In the last decades, these disorders have been models for the development of molecular and cellular therapies for inherited metabolic diseases. Studies in preclinical in vitro systems and animal models have allowed the successful development of bone marrow transplantation (BMT) and enzyme replacement therapy (ERT) as therapeutic options for several LSDs. However, BMT is limited by poor donor availability and high morbidity and mortality, and ERT is not a life-long cure. Moreover, the neuropathology present in many LSDs responded poorly, if at all, to these treatments. Therefore, gene therapy is an attractive therapeutic alternative. Gene therapy strategies for LSDs have employed ex vivo gene transduction of cellular targets with subsequent transplantation of the enzymatically corrected cells, or direct in vivo delivery of the viral vectors. Oncoretroviral vectors and more recently adeno associated vectors (AAV) and lentiviral vectors have been extensively tested, with some success. This review summarises the main gene therapy strategies which have been employed or are under development for both non neurological and neuronopathic LSDs. Some of the in vitro and in vivo preclinical studies presented herein have provided the rationale for a gene therapy clinical trial for Gaucher disease Type I. PMID- 11728221 TI - Histamine dihydrochloride: inhibiting oxidants and synergising IL-2-mediated immune activation in the tumour microenvironment. AB - The potential role of histamine in cancer immunotherapy has been a subject of interest for more than a decade. A significant body of research has elucidated the action of histamine in a model system that mimics the tumour microenvironment. In vitro evidence indicates that histamine inhibits the generation and release of reactive oxygen species (ROS) by monocytes/macrophages (MO) during respiratory burst. Since ROS have been shown to abrogate peritumoural and intratumoural cytokine activation of natural killer (NK) and T-cells and induce apoptosis of these cells in vitro, inhibition of ROS may enable cytokines to activate NK and T-cells and restore their antineoplastic, cytotoxic capabilities. Experimental data indicate that histamine and interleukin-2 (IL-2) act synergistically to activate NK cell cytotoxicity (NKCC). Although IL-2, a regulator of immune responses, has been shown to promote NKCC in monotherapy for metastatic melanoma (MM), renal cell carcinoma (RCC) and acute myeloid leukaemia (AML), objective responses occur in a minority of patients and survival is not significantly extended, except for a minority of patients with MM using high-dose regimens which have not been widely adopted. In vitro findings suggest that the addition of histamine to IL-2 therapy might improve response rates and disease free survival by protecting the cells of the immune system from oxidative stress and inducing natural endogenous immune cytotoxicity. An IL-2/histamine Phase III trial is in progress in a population of AML patients. A recently completed Phase III trial of IL-2 vs. IL-2/histamine in patients with MM demonstrated a trend towards a superior survival benefit from IL-2/histamine for all patients entered, and a statistically significant survival benefit for patients with hepatic metastases. PMID- 11728222 TI - Theratope vaccine (STn-KLH). AB - Active specific immunotherapy (ASI) is a promising approach to treating cancer. Numerous studies in the laboratory have demonstrated that various cancer vaccines can stimulate antibody and cell mediated immune responses against tumour associated antigens [1-9]. Yet few studies have demonstrated convincing clinical responses. Sialyl-Tn (STn) is a carbohydrate associated with the MUC1 mucin on a number of human cancer cells and is associated with more aggressive disease. Consequently, STn is an ideal candidate for ASI therapy. Theratope vaccine is a cancer vaccine that was designed by Biomira, Inc. (Edmonton, Alberta, Canada) by incorporating a synthetic STn antigen that emulates the carbohydrate seen on human tumours. The clinical trials conducted to date with Theratope vaccine are outlined in this report. Overall, Theratope vaccine has been well-tolerated with minimal toxicity. The most common side effects have been in duration and erythema at the site of injections. Both in a non-transplant setting following low dose iv. cyclophosphamide and high dose autologous transplant setting, there has been a trend toward Theratope vaccine decreasing the risk for relapse, prolonging the time to relapse and thus impacting on overall survival. The definitive Phase III trial comparing the outcome of patients with metastatic breast cancer receiving vaccinations with Theratope vaccine versus vaccination with the nonspecific immune stimulants Keyhole Limpet Hemocyanin (KLH) and Detox -B stable emulsion (Detox-B) (now called Enhanzyn Immunostimulant) was closed to enrolment on March 30, 2001. Over 1000 women with distant metastatic breast cancer were enrolled into the program. PMID- 11728223 TI - Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukaemia in first relapse. AB - Gemtuzumab ozogamicin (CMA-676, Mylotarg, an antibody-targeted chemotherapy agent, was recently approved by the FDA for the treatment of patients with CD33+ acute myeloid leukaemia (AML) in first relapse who are 60 years of age or older and who are not considered candidates for other types of cytotoxic chemotherapy. In combined Phase II studies of 142 patients with CD33+ AML in first relapse, gemtuzumab ozogamicin monotherapy was associated with a 30% overall response rate. While treated patients had relatively high incidences of myelosuppression, grade 3 or grade 4 hyperbilirubinaemia (23%) and elevated hepatic transaminases (17%), the incidences of grade 3 or grade 4 mucositis (4%) and infections (28%) were low compared with what might be expected in association with conventional chemotherapeutic treatment. In contrast with the usual in-patient administration of cytarabine and anthracycline-containing induction regimens, a large number of patients were treated with gemtuzumab ozogamicin as outpatients (38% and 41% for the first and second doses, respectively). Two prognostic factors for patients with AML in first relapse, age and duration of complete remission, had relatively little effect on response rates to gemtuzumab ozogamicin. Preliminary data in pediatric patients also suggest the immunoconjugate to be reasonably well tolerated. Studies of gemtuzumab ozogamicin in combination with anthracycline and cytarabine are underway. Gemtuzumab ozogamicin, administered to patients with CD33+ AML in first relapse, has shown overall response rates comparable to conventional agents and a safety profile that appears to be favourable. PMID- 11728224 TI - Recent developments in retroviral gene delivery systems. AB - Since the first gene therapy clinical trial carried out by Rosenberg et al. in 1990 [1], recombinant retroviral vectors are still the most popular gene delivery tools for gene therapy purposes. According to the databases of the Journal of Gene Medicine [101] 35% of gene therapy protocols employ retroviruses, other vectors such as adenoviral vectors are used in 27%, pox viral vectors in 6%, adeno-associated viral vectors in 2% and herpes simplex based vectors in 0.5% of clinical trials. It has become clear that the early retroviral vectors based on simple retroviruses (e.g., murine leukaemia viruses (MLV) are characterised by relatively low viral titre, low transduction efficiency and difficulties in in vivo administration. On the other hand, lentiviral vectors (complex retroviruses) can be grown to a high titre, can stably transduce non-dividing cells and can effectively deliver genes to human progenitor cells (CD34+). This article aims to summarise the first year of the 21st century's developments in the retroviral gene delivery system. PMID- 11728225 TI - Mesenchymal cell transfer for articular cartilage repair. AB - Mature articular cartilage has a poor reparative response to injury and its irreparable breakdown is the common feature of degenerative joint diseases. If articular cartilage lesions become symptomatic, the orthopaedic surgeon must decide on a treatment option. The treatment options include conversion of chondral lesions to osteochondral lesions, which facilitates migration of cells from the marrow space to effect repair. In recent years, a greater emphasis has been placed on tissue engineering strategies and thus several new treatment options have been introduced, including the use of cell transplantation. Several tissue sources and cell types can potentially be used for this type of therapy. These include autologous or allograft chondrocytes and mesenchymal progenitor cells from various tissues. These cells may be delivered to articular cartilage lesions by a variety of methods including direct cell injection to the lesion or seeding in a biodegradable scaffold prior to implantation. In this review, the potential of cell transplantation for articular cartilage repair and regeneration will be discussed. The authors will focus on the available technologies and the present limitations of cell-based therapies. PMID- 11728226 TI - Developments in liposomal drug delivery systems. AB - Liposomes are the leading drug delivery systems for the systemic (iv.) administration of drugs. There are now liposomal formulations of conventional drugs that have received clinical approval and many others in clinical trials that bring benefits of reduced toxicity and enhanced efficacy for the treatment of cancer and other life-threatening diseases. The mechanisms giving rise to the therapeutic advantages of liposomes, such as the ability of long-circulating liposomes to preferentially accumulate at disease sites including tumours, sites of infection and sites of inflammation are increasingly well understood. Further, liposome-based formulations of genetic drugs such as antisense oligonucleotides and plasmids for gene therapy that have clear potential for systemic utility are increasingly available. This paper reviews the liposomal drug delivery field, summarises the success of liposomes for the delivery of small molecules and indicates how this success is being built on to design effective carriers for genetic drugs. PMID- 11728227 TI - Delivery of novel macromolecular drugs against HIV-1. AB - The development of new low molecular weight drugs against human immunodeficiency virus Type 1 (HIV-1) targets other than reverse transcriptase (RT) and protease, such as the integrase and the envelope glycoprotein, is likely to take many years. Macromolecular drugs, including antisense oligonucleotides, ribozymes, RNA decoys and transdominant mutant proteins, may be able to interfere with a relatively large number of viral targets, thereby decreasing the likelihood of the emergence of drug-resistant strains. It may also be relatively easy to alter the sequence of some of the macromolecular drugs to counter emerging drug resistant viruses. The delivery of antisense oligonucleotides and ribozymes to HIV-1 infected or potentially infectable cells by antibody-targeted liposomes, certain cationic lipid formulations and pH-sensitive liposomes results in significant anti-HIV-1 activity. These carriers not only facilitate cytoplasmic delivery but also protect the drugs from nuclease digestion. Delivery of therapeutic genes (another form of macromolecular drug) to target cells is an important challenge of gene therapy. Following delivery by a viral vector, sufficient levels of gene expression must be maintained over an extended period of time to have therapeutic activity. Robust expression of therapeutically useful ribozymes, antisense, decoys and aptamers can be achieved by the use of Pol III expression systems. Moloney murine leukaemia virus- (MoMuLV), adeno-associated virus (AAV)-, or HIV-derived vectors expressing a variety of therapeutic genes have been used successfully to inhibit HIV-1 replication in cultured cells. PMID- 11728228 TI - Gene therapy for rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a disabling, painful disorder affecting 1% of the world's population. Although the aetiology of RA remains unknown, recent advances in understanding its pathophysiology have led to the characterisation of several proteins whose activities may be anti-arthritic. Clinical application of such proteins has greatly improved the treatment of RA, but the disease remains incurable and difficult to manage in a substantial number of patients. Thus, there are continued efforts to develop new therapeutic strategies. Because RA is a chronic condition, effective treatment will probably require the presence of therapeutic agents for extended periods of time. In the case of proteins, this is problematic. Gene therapy may offer a solution to this problem. Experimental studies have confirmed the feasibility, efficacy and safety of gene therapy for the treatment of animal models of arthritis. Several different approaches have shown promise in this regard, including gene transfer to the synovial lining cells of individual joints and the systemic delivery of genes to extra-articular locations. One unexpected finding has been the 'contralateral effect' in which gene delivery to one joint of an animal with polyarticular disease leads to improvement of multiple joints. Investigation of this phenomenon has led to interest in cell trafficking and the genetic modification of antigen-presenting cells (APC). The first Phase I clinical trial tested the feasibility and safety of ex vivo gene transfer to the synovial lining of human joints. This clinical trial has been successfully completed and two other Phase I trials are in progress. A Phase II study is now being planned to investigate the efficacy of gene transfer to the joints of patients with early stage RA. PMID- 11728229 TI - RASONs: a novel antisense oligonucleotide therapeutic approach for asthma. AB - Inhalation based approaches enable the local delivery of antisense oligonucleotides (ASONs) to the respiratory tract and thus facilitate the ability of ASONs to target and modulate the activity of discordantly expressed respiratory disease genes. Studies involving EPI-2010, a respirable antisense oligonucleotide (RASON), targeting the adenosine A(1) receptor, a G-protein coupled-receptor (GPCR) that plays an important role in the aetiology of asthma, demonstrate that ASON therapeutics can be delivered directly to the lung as an aerosol. EPI-2010 has been shown to inhibit adenosine A(1) receptor expression and significantly improve allergen-induced airway obstruction and bronchial hyper responsiveness in animal models of human asthma. Absorption, tissue distribution, metabolism and excretion (ADME) and safety studies of aerosolised EPI-2010 suggest that phosphorothioate RASONs can be delivered to target respiratory tissues in low, safe, efficacious and long-acting doses. This supports the concept that RASONs offer the potential to address a variety of respiratory targets including those for which approaches employing systemic distribution and systemic bioavailability of the therapeutic agent may be undesirable. In addition, our studies with EPI-2010 indicate that the RASON approach may represent a technology that is uniquely positioned to address the challenges of the post-genome era in respiratory drug discovery, since it enables simultaneous in vivo target validation and antisense therapeutic discovery in an accelerated timeframe. PMID- 11728230 TI - Gene transfer to induce angiogenesis in myocardial and limb ischaemia. AB - Stimulation of angiogenesis/arteriogenesis by gene transfer methods offers hope for treating patients with myocardial and peripheral limb ischaemia who are not candidates for standard revascularisation procedures. Preclinical studies showed that adenoviral and plasmid vectors encoding various angiogenic cytokines were capable of inducing functionally significant angiogenesis in vitro and in animal models of chronic myocardial ischaemia. Early clinical studies using VEGF121-, FGF-4- and VEGF165-encoding vectors showed a reasonable safety profile with promising results. However, significant advances in vector technology including regulatable and longer-term expression, delivery strategies (local and organ/tissue specific), clinical trial design, and outcome measure development are needed before this investigational treatment becomes reality. PMID- 11728232 TI - Biological treatment in acute myelogenous leukaemia: how should T-cell targeting immunotherapy be combined with intensive chemotherapy? AB - T-cell targeting immunotherapy is now considered as a possible strategy in the treatment of acute myelogenous leukaemia (AML). Clinical importance of antileukaemic T-cell reactivity after allogeneic stem cell transplantation (SCT) is well established and the early experience from IL-2 therapy suggests that even autologous T-cells can mediate antileukaemic reactivity. The clinical experience also indicates that immunotherapy should begin when the leukaemia cell burden is minimal, and the detection of an operative cellular immune system, even in patients with chemotherapy-induced cytopenia, further suggests that it is possible to begin T-cell targeting therapy early after chemotherapy while patients are still cytopenic. However, adult patients in particular have a T-cell defect after chemotherapy that may last for several months. For this reason immunotherapy should probably be continued or repeated until a maximal effect is achieved when the patients no longer have a T-cell defect. This treatment approach may also be considered in combination with autologous SCT. T-cell targeting regimens should include, if possible, several therapeutic components. Firstly, native AML blasts can function as accessory cells during T-cell activation and in vivo therapy with T-cell growth factors (e.g., IL-2, IL-15) may then enhance antileukaemic reactivity or non-specific cytotoxicity against the AML cells; and secondly, a further enhancement of AML-specific reactivity may be achieved by vaccination with AML-specific peptides, immunisation with AML-blasts expressing a dendritic cell phenotype, or exposure to normal antigen-presenting cells (APC) pulsed with or expressing AML-specific peptide sequences. PMID- 11728231 TI - Antibody targeted therapeutics for lymphoma: new focus on the CD22 antigen and RNA. AB - The approval of antibodies for cancer treatment has provoked increased interest in the development of new and improved antibody-mediated therapies. This emerging approach centres on targeting CD22 on human B-cells with a monoclonal antibody (mAb). Anti-CD22 antibodies conjugated to a cytotoxic RNAse elicits potent and specific killing of the lymphoma cells in vitro and in human lymphoma models in severe combined immune deficiency (SCID) mice. RNA damage caused by RNAses could be an important alternative to standard DNA damaging chemotherapeutics. Moreover, targeted RNAses may overcome problems of toxicity and immunogenicity associated with plant- or bacterial toxin-containing immunotoxins. PMID- 11728233 TI - The treatment, management and prevention of oesophageal cancer. AB - The combination of a rising incidence and a poor survival rate makes oesophageal cancer a major health issue. Adenocarcinoma of the oesophagus is associated with one of the commonest pre-malignant lesions recognised, Barrett's metaplasia. This provides a focus for early detection and intervention. The subjects of acid suppression, bile reflux, COX-2 inhibition and ablation therapy will be discussed herewith. Established carcinoma is now rarely treated by surgery alone and this review discusses the benefits of multimodality therapy combined with more accurate staging techniques. Finally an emerging understanding of the molecular events that characterise the transition to carcinoma may provide novel targets in cancer therapy such as epidermal growth factor receptor (EGFR) and TNF-alpha. This review will focus on some of the future developments in the treatment of oesophageal cancer. PMID- 11728234 TI - Blood stem cell transplantation for breast cancer: new approaches using pre- peri post-transplant immunotherapy. AB - Autologous peripheral blood stem cell transplantation (auto-PBSCT) after high dose chemotherapy is usually offered to breast cancer patients carrying a high risk of relapse or having chemosensitive metastatic disease. Whether progression free and overall survival of such patients is improved after auto-PBSCT compared to conventional chemotherapy is a matter of debate. Currently available results of randomised trials could not uniformly prove or disprove auto-PBSCT being advantageous. Yet such studies have not employed any manipulation of the stem cell graft or any post-transplant immunomodulation exploiting the unique immunological environment for tumour eradication which exists only after auto PBSCT. Preliminary data have discussed the ex vivo and in vivo generation of cytotoxic effector cells employing IL-2 and/or IFN-alpha/gamma in the auto-PBSCT setting. Other cytokines such as IL-12, IL-15 and prolactin have likewise been considered. Several anticancer vaccine protocols after auto-PBSCT are ongoing using monovalent vaccines or anti-idiotypic antibodies. Polyvalent anticancer vaccines, cytokine secreting tumour cells, tumour pulsed or hybridised dendritic cells (DC) enhanced with cytokines are studied. Monoclonal antibodies (mAb) could assist: unlabelled for pretransplant exvivo purging, post-transplant for enhancing antibody-dependent cell mediated cytotoxicity (ADCC) or radioimmunoconjugated as an additive cytotoxic part of the conditioning regimen. Autologous graft versus host induction and allogeneic stem cell transplantation (probably with non-myeloablative conditioning followed by donor lymphocyte infusions) are other approaches. Evaluation of successful combinations, optimal dosages and appropriate timing schedules is the subject of future investigations. Since breast cancer patients belong to countless subgroups, a large number of protocols need to be addressed in order to avoid over treatment and prevent relapse. PMID- 11728235 TI - Omalizumab, a novel anti-IgE therapy in allergic disorders. AB - The incidence of allergic diseases is increasing to epidemic proportions both in the developed and developing world with increasing medical costs and lost productivity. The discovery of immunoglobulin (Ig) E heralded a new era in pathophysiological understanding of allergic disorders. Twenty-five years later, a humanised, non-anaphylactogenic antibody was developed against IgE that could provide a therapeutic alternative to the existing medications. RhuMAb-E25 (omalizumab, Xolair, Genetech, Inc.) is a novel anti-IgE antibody that is directed against the receptor-binding domain of IgE. This binding is specific towards free IgE thereby preventing it from attaching to the mast cell and its subsequent activation. Initial studies demonstrated attenuation of the early and late asthmatic responses when anti-IgE was administered to asthmatic subjects. Later this novel molecule was found to improve symptom scores, rescue medication use, quality of life scores and peak expiratory flows in patients with allergic asthma. Most importantly, omalizumab treatment reduced the corticosteroid use in asthmatic individuals. In patients with seasonal allergic rhinitis, there was a significant reduction in the nasal and ocular symptoms as well as the use of rescue medications. Omalizumab also demonstrated a high level of safety in adults, adolescents and children with a side effect profile no different from the placebo. Its development is an exciting milestone in the treatment of allergic diseases. PMID- 11728236 TI - Alemtuzumab: a novel monoclonal antibody. AB - Antibody-based treatment is a novel, effective management strategy for a variety of diseases. Alemtuzumab (CAMPATH) is an example of a monoclonal antibody (mAb) that was initially developed in the laboratory and has completed its journey by being approved for therapeutic use in humans. The main clinical applications of alemtuzumab include treatment of lymphoid malignancies, particularly chronic lymphocytic leukaemia (CLL) and T-cell prolymphocytic leukaemia (T-PLL) and prevention of graft versus host disease (GVHD) and graft rejection in bone marrow and solid organ transplant recipients. Alemtuzumab administration is accompanied by a characteristic first-dose reaction due to cytokine release that consists of fever, rigors, rash and, at times, dyspnea and hypotension. The most significant toxic effect is profound, prolonged lymphopenia and the subsequent increased risk of opportunistic infections; antibiotic prophylaxis is recommended for patients undergoing alemtuzumab treatment. Alemtuzumab has demonstrated clinical activity as a single agent and has an acceptable toxicity profile. It has significant therapeutic potential, especially against lymphoid malignancies. PMID- 11728237 TI - Effects of wild versus cultivated garlic on blood pressure and other parameters in hypertensive rats. AB - Two separate studies were performed on hypertensive rats to assess the effects of wild, uncultivated garlic on elevated systolic blood pressure (SBP) and other cardiovascular parameters. Also, effects of wild garlic and cultivated garlic preparations were compared and the mechanisms behind pressure-lowering abilities of different garlic preparations were examined. The initial study determined that wild garlic lowers blood pressure. In the second study, cardiovascular effects of three different concentrations of wild garlic and two different cultivated garlics, i.e., a preparation low in allicin and one high in allicin, were compared. All three garlic preparations decreased SBP significantly. Wild garlic produced the greatest pressure-lowering effects, and the least pressure-lowering effects were seen with low-allicin garlic. Compared with control rats, circulating angiotensin II levels were significantly lower in all garlic-eating rats. Losartan decreased blood pressure significantly less and Nw-nitro-L arginine-methyl ester hydrochloride (LNAME) increased blood pressure significantly more in garlic-eating rats than in control rats, suggesting that the renin-angiotensin system (RAS) was less active and the nitric oxide system more active in garlic-consuming hypertensive rats. Accordingly, different garlic preparations, especially wild garlic, favorably influenced high SBP in hypertensive rats. These results suggest that both the RAS and the nitric oxide system are involved in the antihypertensive effects of garlic in hypertensive rats. PMID- 11728238 TI - Effect of risperidone on QT interval and QT dispersion in the elderly. AB - Risperidone, a relatively new antipsychotic medication, is widely used in elderly adults and is reported to have fewer side effects than existing antipsychotic drugs. Recent studies, however, have reported prolongation of QT interval and corrected QT interval, both with the prescribed dose of risperidone and with overdose. Because QT dispersion measured on the surface electrocardiogram (ECG) recently has been shown to be a marker of future arrhythmic events and to predict mortality in elderly patients in a variety of clinical situations, this study was conducted to investigate the effect of risperidone on QT dispersion and corrected QT dispersion in a group of elderly patients. These patients also were evaluated for occurrence of sudden death and/or symptoms suggestive of ventricular arrhythmia. Although risperidone prolonged QT interval, it had no significant effect on QT dispersion. Further, during the follow-up period there were no incidences of sudden death or symptoms suggestive of ventricular arrhythmia. These results indicate that risperidone can be used safely in elderly patients, who are often taking several medications, without risk of increased QT dispersion. PMID- 11728239 TI - Subcapsular hepatic and intraperitoneal bleed after administration of tissue plasminogen activator in a patient with acute myocardial infarction. AB - Thrombolysis can sometimes cause life-threatening bleeding. In the case reported here, subcapsular hepatic and intraperitoneal bleeding developed after administration of tissue plasminogen activator (tPA) in a patient with acute myocardial infarction. Surgery was required in this patient to stop the bleed. PMID- 11728240 TI - Alternative medicine for the conventional cardiologist. AB - Today's cardiologist faces a daily dilemma concerning how best to refer patients for diagnostic procedures and how to select surgical and/or pharmaceutical interventions based on those test results. But the evaluations they order and the treatments they select may actually create unnecessary risks for the patients that are out of proportion to the benefits they experience. Continuing technological advances, although necessary, add to the complexity of the decision making process. As increasing numbers of our patients are turning to alternative therapies, a paradigm shift is required in medicine to evaluate the efficacy of these therapies and effectively incorporate them into the conventional medical model. In this review, alternative treatment options are highlighted, with emphasis on understanding the psychoemotional component of heart disease and the use of modern "nutriceutical" agents. Accepting complementary therapies as equally judicious treatment interventions requires a shift in perspectives that may allow for more integrated care strategies for those who have been seeking a more expanded approach to heart disease. PMID- 11728241 TI - The elderly patient with cardiovascular disease. AB - Cardiovascular disease is a major cause of morbidity and mortality at elderly age. Coronary heart disease (CHD) is the most prevalent problem, followed by hypertensive cardiovascular disease. In this review, the differences in presentation, assessment, management, and prognosis of the common cardiovascular problems in elderly patients compared with younger counterparts are discussed. Principles of cardiovascular drug therapy and issues of preventive care at elderly age also are addressed. Cardiovascular disease in elderly patients is complicated by its superimposition on the physiologic and structural cardiovascular changes of aging and by its frequent association with multiple other comorbid illnesses. Most diagnostic and therapeutic procedures are characterized by an excess of adverse responses compared with those in patients of younger age. The challenge in caring for the escalating elderly population in the twenty-first century will be to delineate those interventions that favorably affect morbidity and mortality, functional status, and quality of life attributes. PMID- 11728242 TI - Drug treatment of peripheral vascular disease. AB - There are a limited number of clinically effective pharmacotherapeutic agents for treatment of peripheral vascular disorders. Pentoxifylline and cilostazol are available for the symptomatic treatment of intermittent claudication. Analogs of carnitine and L-arginine are being evaluated for treatment of the symptoms of intermittent claudication, and prostaglandins and growth factors are being evaluated for critical limb ischemia. Calcium channel blockers remain the treatment of choice for Raynaud's phenomenon. Alternative vasodilators may be used selectively to treat individuals with Raynaud's phenomenon who are intolerant of calcium channel blockers or in whom such therapy has been unsuccessful. Prostaglandins have been evaluated in patients with refractory Raynaud's phenomenon who also have digital ulceration. The mainstay of short-term treatment (and prophylaxis) of deep vein thrombosis (DVT) has been unfractionated heparin, but now the use of low molecular weight heparin (LMWH) has emerged. Newer agents, such as heparinoids and direct thrombin inhibitors, hold promise for the prevention and treatment of DVT. Prolonged treatment with warfarin is still required to prevent recurrent thrombosis, although the duration of treatment has come under debate. PMID- 11728243 TI - Major depression as a risk factor for cardiovascular disease: therapeutic implications. AB - This review was conducted to analyze the literature regarding the association between psychological depression and cardiovascular disease (CVD). The literature was reviewed through Medline and Psychlit. Articles were located using search terms such as depression, CVD, coronary artery disease, and cardiac risk factor. The search included articles from 1980 through 1999. Both the cross-sectional and prospective data suggest that psychological depression is a risk factor for and an aggravating factor of preexisting CVD. Proposed pathophysiologic mechanisms and therapeutic implications are discussed. PMID- 11728244 TI - Imidazoline receptor agonist drugs for treatment of systemic hypertension and congestive heart failure. AB - The imidazoline receptors recently have been discovered to be involved in the central nervous system control of sympathetic outflow. A new class of centrally acting antihypertensive agents, the imidazoline receptor agonists (rilmenidine and moxonidine), have been developed to control blood pressure effectively without the adverse effects of sedation and mental depression that usually are associated with centrally acting antihypertensive agents. This new generation of centrally acting antihypertensive agents is highly selective for the imidazoline receptor but has a low affinity for alpha(2)-adrenergic receptors. The usefulness of these agents in the treatment of congestive heart failure has not been demonstrated. PMID- 11728245 TI - Cerivastatin. AB - Cerivastatin is a synthetic and enantiomerically pure 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor. It has been recognized for its high pharmacologic potency, uncomplicated pharmacokinetic profile, and low drug interaction potential. The efficacy of cerivastatin has been demonstrated in a number of clinical trials involving patients with primary hypercholesterolemia and mixed dyslipidemia. Cerivastatin was found to have dose-dependent reduction in total cholesterol and low-density lipoprotein (LDL) cholesterol. At the 0.4 mg daily dose, cerivastatin was found to lower LDL cholesterol by approximately 35% (mean reductions of 33-39%). The triglyceride-lowering effect of cerivastatin is also dose dependent; however, there is a much stronger association between baseline triglyceride levels and the triglyceride-lowering effect of cerivastatin. A dosage of 0.4 mg daily has been shown to reduce triglycerides by a mean of 28% in patients with baseline triglyceride levels of > 300 mg/dL. At the time of its initial approval by the US Food and Drug Administration (FDA) in 1997, cerivastatin was available only in 0.2 mg and 0.3 mg strengths. The recent FDA approval of the 0.4-mg strength dose has made cerivastatin a more competitive drug in lipid-lowering efficacy among the HMG-CoA reductase inhibitors. PMID- 11728246 TI - Matched comparison of slotted tubular and coil stents: differences in acute gain, loss index, and clinical outcome. AB - Different stent designs may produce differences in immediate and long-term outcome. The objective of this study was to compare the immediate results and long-term outcomes of patients with lesions undergoing implantation of a single slotted tubular stent (Palmaz-Schatz) or a single coil stent (Gianturco-Roubin I). For this purpose, the authors studied patients matched for lesion length and vessel size. The consisted of 135 patients (141 lesions), and the Gianturco Roubin group was composed of 50 patients (56 lesions). The coil stent was more often used for bailout indications. Larger immediate lumen gain was achieved with the slotted tubular stent. At follow-up examination, there was no difference in stent thrombosis; however, there was a trend toward lower angiographic restenosis and target-lesion revascularization in the Palmaz-Schatz group. Randomized trials using larger numbers of patients are needed to determine differences in outcome among different stent designs. PMID- 11728247 TI - Cardiomyocyte aging is gender-dependent: the local IGF-1-IGF-1R system. AB - To determine whether insulin-like growth factor 1 (IGF-1) and its receptor (IGF 1R) are implicated in the aging process of the heart, and if their impact differs in the two genders, the expression of IGF-1, and extracellular alpha-subunit and transmembrane beta-subunit of IGF-1R was measured in left ventricular myocytes isolated from male and female Fischer 344 rats at 3, 8, 12, 16, and 26 months after birth. Additionally, the extent of myocardial damage in both sexes was evaluated in rats at 3 and 26 months by confocal microscopy. Finally, ventricular hemodynamics was assessed in the closed-chest preparation. From 3 to 26 months, aging was characterized by an 83%, 84% decrease and disappearance in the quantity of IGF-1, IGF-1Ralpha and IGF-1Rbeta in male myocytes. Corresponding changes in female myocytes were 40%, 28% and 43%. These molecular modifications at the myocyte level were coupled with tissue injury, consisting of multiple foci of replacement fibrosis across the left ventricular wall. However, myocardial fibrosis in females was 76% and 77% significantly less than in the young and old male heart, respectively. These multiple age-associated events were accompanied by cardiac decompensation in the senescent male rat, while modest indices of ventricular dysfunction were detected in old female rats. In conclusion, the enhanced IGF-1-IGF-1R system in female myocytes may condition the favorable outcome of age in this gender. PMID- 11728248 TI - Hypoxemia sans hypoxemia. AB - Acquired methemoglobinemia usually occurs during endoscopy after exposure to a variety of drugs commonly used in critical care units and in endoscopy suites. A rapid infusion of methylene blue can result in a complete recovery of patients with a favorable prognosis. The authors describe a case of 31-year-old woman in whom methemoglobinemia developed with the use of benzocaine, and who showed complete recovery after methylene blue infusion. Hence, the availability of methylene blue in these settings is advocated. PMID- 11728249 TI - Current thoughts on the management of renovascular disease. AB - Renal artery stenosis has long been recognized as a cause of systemic hypertension, and has more recently been identified as a cause of progressive renal insufficiency. The diagnosis of renal artery stenosis often has been overlooked because of the difficulty in establishing the diagnosis. Until recently, surgical intervention has been the only therapeutic modality available. This article will review some recent advances in imaging modalities, particularly duplex ultrasound and magnetic resonance angiography, which have allowed easier visibility of the renal vessels. Second, the introduction of renal percutaneous transluminal angioplasty, with and without stent placement, has made operative intervention more attractive. The early and short-term results of this procedure are discussed and compared with surgical experience. Finally, the implications for using these newer interventional techniques for minimally symptomatic renal artery stenosis are reviewed. PMID- 11728250 TI - Pharmacologic approaches for the management of systemic hypertension in pregnancy. AB - Hypertension in pregnancy includes a group of distinct disorders that require special consideration in both prevention and pharmacologic treatment. In recent years, there have been few advances regarding the pathophysiology and prevention of preeclampsia, or in recommendations for first-line drug therapy of the hypertensive complications of preeclampsia. Similarly, the recommendations for pharmacologic treatment of women with chronic hypertension antedating pregnancy have changed little, primarily because first-line medications have the advantage of having been the subjects of extensive research experience. Recent clinical trials have demonstrated the efficacy and safety of various second-line drugs for treating hypertensive disorders of pregnancy; whether these therapies can eventually replace the standard recommended medications will require more extensive long-term investigation. PMID- 11728251 TI - Cardiac complications of human immunodeficiency virus infection: diagnostic and therapeutic considerations. AB - Cardiac disease is being recognized as a complication of human immunodeficiency virus (HIV) infection, as more effective therapy is producing longer survival in patients with HIV infection and acquired immune deficiency syndrome (AIDS). Cardiac disease may occur coincidentally in a patient with AIDS, as a complication of AIDS, as a result of therapy for AIDS, or even as a direct result of HIV infection of the heart. Congestive heart failure (CHF) carries a poor prognosis and is best treated with traditional therapy. All patients with HIV/AIDS should get a baseline echocardiogram and electrocardiogram (ECG), because many patients are asymptomatic. Patients with low CD4 counts, those receiving zidovudine, and intravenous drug users must be further evaluated. Most patients with symptomatic effusions have a potentially treatable cause (neoplasm or infection), and a full workup must be initiated; however, small asymptomatic effusions often can be observed and followed by serial echocardiography. Physicians should be more alert to cardiac involvement in patients with HIV. Earlier surveillance is warranted and may lead to earlier treatment and supportive care. PMID- 11728252 TI - Esprolol hydrochloride: a new beta adrenergic antagonist with a rapid onset of effect. AB - Esprolol is a new, potent, beta adrenergic receptor antagonist that is differentiated from available drugs of this class by its rapid onset, short time to peak effect, and relatively short duration of action after sublingual and oral administration. The drug has potential for short-term use in patients with exertional angina and in patients with other diseases in which rapid-onset beta blocking action would be beneficial. The unique properties of esprolol are determined by the presence in its chemical structure of an ester group, which undergoes rapid metabolism by blood and tissue esterases to form an active metabolite, amoxolol. Preclinical testing and clinical trials in healthy volunteers have demonstrated the tolerability, pharmacokinetics, and beta adrenergic blocking properties of both esprolol and amoxolol. An ongoing trial to evaluate its usefulness in improving exercise performance in patients with exertional angina pectoris is nearing completion. PMID- 11728253 TI - Cardiovascular drug therapy in the elderly. AB - Cardiovascular diseases of various etiologies are most prevalent in the elderly, and there are many pharmacologic issues that need to be considered when prescribing drug therapies to this population. Aging itself alters the absorption, distribution, metabolism, and excretion of many drugs. Certain drugs, such as digoxin, lidocaine, and warfarin, need to be used with great caution in older patients because of a greater potential for drug toxicity. Elderly patients often are prescribed multiple drugs for different conditions, making them susceptible to major drug-drug interactions. Recommendations are provided to help ensure safe prescribing practices of cardiac drugs in older patients. PMID- 11728254 TI - Clopidogrel: a new antiplatelet drug. AB - Clopidogrel is a new drug in the recently developed class of thienopyridine derivatives that inhibits platelet function by an inhibitory action exerted through the membrane adenosine diphosphate receptor. Clopidogrel is a prodrug that must be metabolized to an active metabolite in the liver. The basic chemistry, pharmacodynamics and pharmacokinetics of the drug are reviewed. Clinical trials with clopidogrel, including its use in patients with stents postangioplasty, adverse reactions, and potential advantages over other agents are summarized. PMID- 11728255 TI - Orlistat. AB - Obesity is a risk factor for cardiovascular disease. Orlistat is a gastric and pancreatic lipase inhibitor indicated for the management of obesity. It is the first antiobesity agent that is not a centrally acting appetite suppressant; instead, it decreases absorption of dietary fat in the gastrointestinal tract. The effects of orlistat on weight loss, weight regain, and on a number of obesity related risk factors have been assessed in large clinical trials of 1 to 2 years' duration. Compared with subjects who received placebo and a hypocaloric diet alone, weight loss of at least 5 to 10% of initial body weight was observed in a significantly larger number of subjects who were treated with orlistat plus a hypocaloric diet during the first year of treatment. Subjects who received orlistat 120 mg three times daily regained significantly less weight than subjects who received placebo during the second year of treatment. In addition, orlistat was found to have favorable effects on blood pressure and concentrations of serum lipid, glucose, and insulin. Gastrointestinal events are the most common adverse effects experienced by patients who received orlistat; however, most of these events were mild to moderate in intensity, transient in duration, and decreased considerably during the second year of treatment. PMID- 11728256 TI - ACE inhibition and vascular remodeling of resistance vessels: vascular compliance and cardiovascular implications. AB - The arterial circulation is composed of conduit arteries that store blood in systole (capacitive function), small branch points that contribute to pressure oscillations and reflected waves (oscillatory function), and arterioles that control blood flow (resistance function). These vessels are lined by endothelial cells that secrete nitric oxide and other substances that influence smooth-muscle tone, growth, and structural remodeling. The role of endothelial function is greater in thin-walled vessels than in conduit arteries, including proximal elastic vessels with thick walls. Thus, endothelial dysfunction is most likely to raise resistance and alter pressure oscillations than to produce early changes in the conduit artery wall. Later structural changes, facilitated by endothelial dysfunction, lead to reduced large artery compliance and widened pulse pressure as markers of risk for a cardiovascular event. Angiotensin-converting enzyme inhibitors, particularly those with vascular effects, tend to restore endothelial function and may result in regression of the structural alterations. Pulse contour analysis is the most sensitive means of assessing both capacitive oscillatory and resistance vessel function of the vasculature. Noninvasive radial artery tonometry with computer analysis of diastolic pressure decay provides independent measures of capacitive function, oscillatory function, and resistance, which allows screening and monitoring of arterial function. Such monitoring should lead to more individualized management that could supplement pressure monitoring as a guide to the efficacy of therapeutic intervention. PMID- 11728257 TI - Evaluation of once-daily antihypertensive therapy. AB - The sixth report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of Hypertension report has recommended long-acting, once daily drugs as preferred therapy for the management of hypertension. The duration of action of an antihypertensive agent can be assessed in several ways. The decrease in clinical blood pressure at the end of the dosing interval has traditionally been the method for determining antihypertensive efficacy. However, this approach does not provide information about a drug's effects over the remainder of the 24-hour dosing interval. The trough-to-peak ratio has been proposed as an arithmetic index of a compound's maximum and minimum antihypertensive effects. This ratio can be obtained from multiple readings in an office setting or using data from 24-hour ambulatory blood-pressure monitoring. A simplified approach has been to compare a single office reading at the time of a drug's known maximum antihypertensive effect (peak) with the reduction in blood pressure at 24 hours (trough). Office readings have also been used to show that the same dose of a drug given once daily versus twice daily results in a similar trough blood pressure, confirming that once-daily therapy is appropriate. The 24 hour ambulatory blood pressure can be used to determine mean changes in daytime, evening, and night-time blood pressure to demonstrate the antihypertensive profile over the entire dosing interval. Finally, the finding of a persistent reduction in ambulatory blood pressure following substitution of placebo for an antihypertensive drug during chronic therapy (missed-dose technique) suggests that a compound decreases blood pressure for 48 hours or longer. Thus, it should be possible to demonstrate the effectiveness of once-daily antihypertensive therapy using one or more of the aforementioned methods. PMID- 11728258 TI - Global impact of stroke. AB - As the developing world becomes increasingly industrialized and urbanized, and its vast populations adopt more westernized diets and lifestyles, the burden of cardiovascular disease and disability is growing at an alarming rate. The results of prospective observational studies conducted in both western and eastern populations have established the strong association between the primary incidence of stroke and the level of blood pressure. Further, randomized clinical trials of antihypertensive therapy have demonstrated a reduction in primary stroke risk. Among persons with a history of cerebrovascular disease, data from the United Kingdom Transient Ischemic Attack Aspirin Trial (UKTIA) has demonstrated a similar relationship between blood pressure and secondary stroke risk. Trials of blood-pressure lowering in patients with a history of cerebrovascular disease also suggested a reduction in stroke risk. However, patients numbers in these trials were small, and the results were not conclusive. PROGRESS (Perindopril pROtection aGainst Recurrent Stroke Study) is a large, randomized, double-blind, controlled trial designed to provide a definitive answer to these questions. Its primary aim is to determine the effect of blood-pressure lowering on stroke incidence in patients with a history of a previous stroke or transient ischemic attack. A total of 6,105 patients were randomized to receive perindopril (plus indapamide) or placebo in seven regions of the world. The current mean follow-up period exceeds 30 months, and a total follow-up period of at least 4 years is planned. Secondary outcomes include the incidence of other major cardiovascular events, such as myocardial infarction, and of cognitive decline and dementia. Both the major event rate and the degree of adherence to randomized therapy exceed initial projections, which suggests that the study should have the power to achieve its main objectives by the planned completion date of 2001. PMID- 11728259 TI - Cardiorenal protection in diabetes. AB - Diabetes is twice as common in hypertensive patients than in the general population, and is a major cause of cardiovascular morbidity. Diabetes is the most common cause of end-stage renal disease in the United States, and is primarily responsible for the 9% increase in prevalence of end-stage renal disease during the past 10 years. However, there is evidence that tight blood pressure control can reduce the vascular complications of diabetes. This reduction was demonstrated in the United Kingdom Prospective Diabetes Study, in which patients who were randomized to a tight blood-pressure control group had 24% fewer vascular complications (including strokes and diabetic retinopathy) than patients in usual-care groups. Thus, the target goal for blood pressure in patients with diabetes is 130/85 mmHg. Microalbuminuria is an early marker of diabetic nephropathy, and is most pronounced in patients whose 24-hour blood pressure shows a nondipping pattern (failure of the blood pressure to fall at night). The prevalence of nondipping blood pressure is increased in patients with diabetes and in patients with renal disease due to other causes. Further, there is evidence that patients with nondipping blood pressure show more rapid deterioration of renal function than patients with dipping blood pressure. High nocturnal pressure in patients with nondipping blood pressure may accelerate glomerular damage. For example, studies of animals with diabetes have shown that the afferent glomerular arteriole is dilated when compared with the efferent arteriole, thereby exposing the glomerulus to the systemic arterial pressure. Numerous studies have shown that angiotensin-converting enzyme (ACE) inhibitors are superior to some antihypertensive agents (particularly beta blockers and diuretics) in the prevention of renal damage. This observation is based on results of a metaanalysis that demonstrated a 9%/year glomerular filtration rate decrease in patients taking beta blockers or diuretics, but only a 1%/year decrease in patients taking ACE inhibitors. The role of calcium channel blockers is more controversial, but there is evidence to suggest that nondihydropyridine calcium channel blockers may confer the same benefit as ACE inhibitors. Alpha blockers and angiotensin-receptor blockers show promise, but have been less widely studied. PMID- 11728260 TI - Aspirin does not interact with ACE inhibitors when both are given early after acute myocardial infarction: results of the GISSI-3 Trial. AB - Aspirin (ASA) and angiotensin-converting enzyme inhibitor (ACEi) therapy reduce mortality when administered early after the onset of myocardial infarction. ASA can antagonize some effects of ACEi therapy by inhibiting the synthesis of vasodilating prostaglandins; however, the evidence for this effect from large controlled trials is contradictory. The authors analyzed a database of 18,895 patients of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardio-3 (GISSI-3) Trial in which patients were allocated either to receive lisinopril or not to receive lisinopril within 24 hours of the onset of symptoms of myocardial infarction. The aim of the study was to verify the possible negative interaction between ASA and the ACEi lisinopril in the postacute phase of acute myocardial infarction. Of 18,895 analyzable patients, 15,841 received ASA at entry. Overall lisinopril reduced 42-day mortality from 7.1% to 6.3%. In patients receiving ASA, mortality was reduced by lisinopril from 6.0% to 5.4%, and from 13.0% to 10.8% in patients not receiving ASA. The difference in proportional reductions of mortality corresponds to the fact that a more marked lisinopril effect is seen in patients at higher baseline risk across all study subgroups, one of which coincides with the no-ASA group. The analysis of the inhospital incidence of major clinical events did not reveal a potentially negative interaction between ASA and lisinopril. The same findings were obtained from the analysis of reinfarction at 42 days. The interaction between ASA and lisinopril was also tested by multivariate analysis adjusted for confounding variables at entry, and the interaction tests were not statistically significant. Serum creatinine levels at 42 days were significantly higher in lisinopril group than in the control group. Systolic and diastolic blood pressures in lisinopril group were significantly lower than controls at 42 days. The effect of lisinopril on creatinine and blood pressure did not differ between the ASA and no-ASA groups. ASA does not decrease the mortality benefit of early lisinopril after myocardial infarction, nor does it increase the risk of major adverse events. Lisinopril is safe and effective when given early after the onset of myocardial infarction, regardless of a concomitant administration of ASA started early and continued over a 6-week period. PMID- 11728261 TI - Preconditioning myocardium with demand ischemia in the presence of a critical coronary artery stenosis. AB - The relative importance of adrenergic stimulation and demand ischemia as important preconditioning stimuli remains unclarified. The purpose of this investigation was to use a partial coronary stenosis to define the preconditioning role of demand ischemia. Dobutamine was infused intravenously before coronary occlusion in closed-chest swine with and without an artificial coronary stenosis in the mid-left anterior descending coronary artery. Control animals had no stenosis and did not receive dobutamine before occlusion. All three groups underwent 45 minutes of occlusion followed by 120 minutes of reperfusion. At baseline, regional myocardial blood flow in the area at risk was reduced in animals with a stenosis, but global left ventricular systolic function, measured by gated blood pool scan, was equivalent in all three groups. Animals with and without a stenosis received equivalent catecholamine stress with dobutamine, but only animals with a stenosis manifested ischemia during the infusion. At 2 hours after reperfusion, infarct size as a percentage of the area at risk was smaller in animals with a stenosis given dobutamine. Demand ischemia preconditions myocardium in closed-chest swine. Increased demand alone without ischemia had marginal preconditioning effects. This may have clinical relevance to patients with severe stenoses exposed to stressful stimuli before the development of myocardial infarction. PMID- 11728262 TI - Health-related quality of life in the treatment of angina. AB - The objectives of this study were to review the literature on health-related quality of life (HRQOL) in patients with angina pectoris (AP), provide suggestions for future research, and propose practical considerations for the selection and use of HRQOL instruments in AP clinical trials. A MEDLINE search was conducted of literature published between January 1980 and April 1999 using MeSH terms "quality of life" and "heart disease." Sixty articles were eligible for inclusion. General and disease-specific instruments were used in 55% and 57%, respectively. Findings were inconsistent with respect to which disease-specific or generic aspects of HRQOL were affected by treatment or which measures were sensitive to treatment changes. Disease-specific measures were more sensitive to treatment than general measures. Evidence suggests that HRQOL effects of surgical and medical treatments are similar over the long term. HRQOL is an important outcome measure to consider. Both disease-specific and generic measures should be used, and a variety of dimensions should be addressed when evaluating HRQOL of patients with AP. Further research is needed to assess the long-term effect of various treatments on HRQOL and the predictive power of HRQOL in clinical outcomes. PMID- 11728263 TI - Recurrent right atrial thrombus after repair of ruptured sinus of valsalva aneurysm. AB - Ruptured sinus of Valsalva aneurysm is a rare cardiac abnormality. Early surgical repair is indicated to prevent complications such as heart failure, infective endocarditis, arrhythmias, and thromboembolic events. The authors report an unusual case of recurrent right atrial thrombus after the surgical repair and closure of ruptured sinus of Valsalva aneurysm. PMID- 11728264 TI - Adverse dermatologic effects of cardiovascular drug therapy. AB - Cardiovascular disease is common, affecting an increasing number of persons as the population ages. To combat this growing health problem, physicians use a multitude of medications in the treatment of their patients. Although pharmacologic therapy greatly enhances quality of life for a majority of patients, there is always the potential for an unfavorable reaction. For example, cardiovascular drugs can induce a vast array of adverse dermatologic responses. This article reviews the various cutaneous reaction patterns that can occur as a result of treatment with specific cardiovascular agents. PMID- 11728265 TI - Left ventricular reduction surgery. AB - When medical therapy is unsuccessful in improving the functional class in patients with heart failure, surgical options must be considered. Although transplantation may be considered the gold standard, numerous factors limit the number of patients who can benefit from it. Therefore, other surgical methods have been devised as alternatives to transplantation. Left ventricular reduction by surgical resection of a piece of myocardium was introduced in Brazil a few years ago, and enthusiasm for the procedure is increasing because encouraging results have been published by various groups. It is hoped that this procedure will form part of the treatment for congestive heart failure in the future. PMID- 11728266 TI - Cytochrome p450-mediated cardiovascular drug interactions. AB - Numerous cardiovascular agents are substrates, inhibitors, or inducers of cytochrome P450 (CYP450) enzymes. The growing understanding of the CYP450 enzyme family allows us to predict potential drug interactions. This review discusses CYP450-mediated cardiovascular drug interactions with selected examples reported in the literature. The principles of predicting drug interactions are also discussed. Studies evaluating in vivo drug interaction and case reporting by clinicians are necessary to establish the clinical significance of these interactions, which will in turn help to develop strategies to prevent and manage the potentially devastating effects they might have on patients. PMID- 11728267 TI - Adrenomedullin: a vasoactive and natriuretic peptide with therapeutic potential. AB - Adrenomedullin is a potent endogenous vasodilating and natriuretic peptide that is similar in structure to calcitonin gene-related peptide (CGRP). The gene involved in the synthesis of adrenomedullin has been localized to a single locus on chromosome 11, with specific sites on the genome to regulate transcription. Adrenomedullin is normally found in human plasma and in other organs. It is thought that one of the clearance sites for this peptide is in the pulmonary circulation. Endothelial cells are assumed to be one of the major sources of plasma adrenomedullin. Adrenomedullin is an important factor in regulating local and systemic vascular tone, by its activity as an autocrine/paracrine and circulating hormone. Depending on the site of action, adrenomedullin seems to bind to a CGRP receptor and send signals by either cyclic adenosine monophosphate or nitric oxide. From the results of experiments in animals, it has become clear that adrenomedullin's effects are species-specific. However, what is commonly seen with adrenomedullin is peripheral vasodilatation, a positive inotropic action, increased cardiac output, and increased stroke volume. In addition, adrenomedullin has actions in the brain, lungs, and kidneys to regulate regional hemodynamics. With these activities defined, recent studies have suggested a potential therapeutic role for adrenomedullin. PMID- 11728268 TI - Omapatrilat: a unique new agent for the treatment of cardiovascular disease. AB - Omapatrilat is the most clinically advanced of a new class of drugs, vasopeptidase inhibitors, which are being studied for the treatment of patients with cardiovascular disease. Omapatrilat inhibits the enzymatic activities of angiotensin-converting enzyme and neutral endopeptidase. The end result is blockade of angiotensin-II formation and inhibition of the catabolism of vasodilatory hormones, such as the natriuretic peptides, bradykinin, and adrenomedullin. Some of the ultimate pharmacologic effects include vasodilation, natriuresis, and diuresis, which may be beneficial in the management of various cardiovascular diseases, such as hypertension and heart failure. The pharmacokinetics of omapatrilat are compatible with once-daily dosing and a duration of antihypertensive efficacy of more than 24 hours. Omapatrilat decreases blood pressure in both high-renin and low-renin states, which suggests antihypertensive efficacy that is independent of the status of the renin angiotensin system. Furthermore, the antihypertensive effect of omapatrilat is indiscriminate of age or race. Omapatrilat has consistently shown efficacy in decreasing both systolic and diastolic blood pressure to a similar or greater extent than either lisinopril or amlodipine; however, systolic pressure is more responsive to omapatrilat treatment than diastolic pressure. Although the role of omapatrilat in heart failure is still evolving, preliminary results are promising: hemodynamic improvements and clinical benefits of omapatrilat are similar or greater to those achieved with an angiotensin-converting enzyme inhibitor. Future studies (specifically the OVERTURE Study) will be of pivotal importance in establishing the role of omapatrilat in the treatment of patients with heart failure. The side-effect and drug-interaction profiles of omapatrilat are largely incomplete, but suggest excellent tolerability and a side-effect profile that is similar to placebo. Omapatrilat could be a revolutionary addition to the management of cardiovascular disease, and its clinical development will be followed closely by many who are curious if larger clinical trials will echo the impressive preliminary data that have been seen thus far. PMID- 11728269 TI - Comparison of subcutaneous calcium heparin and acetylsalicylic acid in the prevention of ischemic events and death after myocardial infarction: a randomized trial in a consecutive series of 90 patients. AB - This study compares the prophylactic effectiveness of calcium heparin (CH) with that of acetylsalicylic acid (ASA) in the long-term anticoagulant management of patients with myocardial infarct. Forty-four patients discharged from a coronary care unit after acute myocardial infarction were randomly assigned to receive 12,500 IU subcutaneous CH daily for 3 months, followed by 325 mg ASA daily for 3 months. This protocol was compared with the administration of 325 mg/day ASA to an additional 46 patients for 6 months. There were 16 dropouts in the CH group and two in the ASA group. Ischemic events, surgery, and adverse effects totaled 36 in the CH group versus 73 in the ASA group. Three patients from each group died (6.8% vs. 6.5% in the CH and ASA groups, respectively). Patients in the CH group were significantly older (76 +/- 3.51 years vs. 63 +/- 5.51 years in the CH and ASA groups, respectively; P = 0.026). Although ASA is generally regarded as the most effective way of preventing the recurrence of ischemic events, the results of this study suggest that CH is a potentially useful and well-tolerated alternative that could be employed when ASA is contraindicated. PMID- 11728270 TI - Percutaneous transluminal coronary angioplasty (PTCA) combined with stenting improves clinical outcomes compared with PTCA alone in acute myocardial infarction. AB - Balloon angioplasty in acute myocardial infarction is an established procedure. The procedure is limited by the potential for early abrupt reocclusion (18-20%) and other complications. Coronary stenting improves the initial success rate, decreases the incidence of abrupt closure, and is associated with a reduced rate of restenosis. For these reasons, coronary stenting is increasingly utilized to treat acute myocardial infarction. The purpose of this study was to compare the effect of coronary stenting with percutaneous transluminal coronary angioplasty (PTCA) in the management of acute myocardial infarction. Clinical data from a retrospective review of 228 consecutive patients admitted with acute myocardial infarction who underwent primary or rescue coronary intervention were used. There was a significantly greater incidence of in-hospital recurrent myocardial infarction in the PTCA group (10%) versus the stented group (1%). In the PTCA cohort, 10 patients required in-hospital repeat revascularization by PTCA compared with one patient in the stented group. The in-hospital death rate was significantly higher in the PTCA group (8%) compared with the stented group (1%). There was no significant difference in the incidence of postdischarge death or repeated revascularization. The results suggest that patients who undergo PTCA with stent deployment have fewer episodes of in-hospital recurrent angina, myocardial infarctions, repeat angioplasties, and fewer in-hospital deaths. PTCA with stenting was associated with a low in-hospital mortality (1%). For patients who had PTCA alone and survived to be discharged, there was no significant difference in overall mortality or the need for revascularization over the 2-year follow-up period. PMID- 11728271 TI - Clinical research opportunities: what you need to know for your practice. AB - Cardiologists and cardiovascular practitioners are facing significant reimbursement constraints. In this setting, increasing numbers of practitioners are looking to participate in clinical research as a new source of revenue. An aging population and consolidation among pharmaceutical companies, medical device companies, and contract research organizations are placing great emphasis on speed of new products to market. Opportunities exist for well-positioned and knowledgeable clinical investigators to enhance their practices, increase their professional knowledge, and expand their network of scientific colleagues. This article provides an overview of the drug and pharmaceutical industry referable to clinical investigation, the tools necessary to become a valued, principal investigator and an outline of challenges and opportunities for clinicians wishing to engage and enhance their clinical research activities. PMID- 11728272 TI - A case of a large coronary artery fistula in an elderly patient surgically corrected 12 years after initial diagnosis. AB - This case report discusses the natural history of a large coronary artery fistula in an elderly patient treated conservatively for 12 years. There has been no previous report of long term follow-up in a patient with a large coronary artery fistula with symptoms of congestive heart failure. The surgical management and clinical response to surgical correction will be discussed, and an extensive review of the literature will be performed. PMID- 11728273 TI - Use of endovascular stents for the treatment of abdominal aortic aneurysm. AB - The conventional open surgical repair of abdominal aortic aneurysms is a well established and durable procedure. However, as with all other major abdominal surgical operations, there are associated morbidities. Both mortality and morbidity increase significantly with advanced patient age and associated comorbid conditions. Endovascular aortic stent-grafts offer a significantly less invasive alternative to conventional open-surgical repair. Patients previously considered unsuitable for open repair can often receive treatment for aneurysms with endovascular techniques. The US Food and Drug Administration has recently approved two endovascular stent-graft devices for use within the United States. PMID- 11728274 TI - Prevention of sudden cardiac death. AB - Tremendous strides have been made in recent years in the treatment and prevention of sudden cardiac death. Large-scale trials have identified several strategies that may be implemented to improve survival in patients susceptible to sudden cardiac death. In patients who have had a sustained ventricular tachyarrhythmia, the current therapy of choice is an implantable cardioverter defibrillator. For prophylaxis of sudden cardiac death in patients without a previous event, several approaches should be considered. Beta-blockers are an effective pharmacologic therapy in patients after myocardial infarction, and their efficacy has also been demonstrated in patients with congestive heart failure. Antiarrhythmic drugs are not efficacious as prophylactic agents to reduce mortality in these populations. The best therapeutic approach for prophylactic therapy to prevent sudden cardiac death appears to be the implantable cardioverter defibrillator; however, its use can be justified only in patients at high risk for developing sudden cardiac death. Further work is needed to identify the high-risk populations in which this therapy is warranted. PMID- 11728275 TI - Therapeutic angiogenesis: a new treatment modality for ischemic heart disease. AB - Angiogenesis is the process of new blood vessel formation, and has potential clinical use in the management of ischemic heart disease. A considerable amount of ongoing research has recently focused on the process of angiogenesis, including the identification of various factors that can inhibit or stimulate this process. The picture that is emerging suggests that a complex set of interactions involving various cells and cellular products is the key to angiogenesis. In particular, endothelial cells and growth factors, such as vascular endothelial growth factor and fibroblast growth factor, appear to play integral roles in angiogenesis. Various preclinical studies involving animal models of ischemia explored the potential use of angiogenesis in ischemic disease. Based on encouraging results, a number of clinical trials involving angiogenesis have been initiated to determine whether the process of angiogenesis also offers therapeutic benefit in humans. PMID- 11728276 TI - Rosiglitazone: an agent from the thiazolidinedione class for the treatment of type 2 diabetes. AB - Great advances have been made in the management of diabetes during the past decade. Whereas only one class of oral medications (the sulfonylureas) was available for the treatment of type 2 diabetes in the early 1990s, we now have five classes of oral antidiabetic agents from which to choose. The thiazolidinedione class of medications was first introduced to the United States when troglitazone was marketed during early 1997. Rosiglitazone, approved by the FDA during the spring of 1999, was the second thiazolidinedione to be marketed in the United States. Similar to troglitazone, rosiglitazone improves insulin sensitivity in patients with type 2 diabetes by activating peroxisome proliferator-activated receptor-gamma (PPARgamma) receptors in adipose tissues, skeletal muscles, and the liver. The efficacy and safety of rosiglitazone therapy in patients with type 2 diabetes have been demonstrated in a number of clinical studies, which are summarized in this article. Selected characteristics of rosiglitazone are compared with those of pioglitazone--the other thiazolidinedione currently available in the United States. Edema of mild to moderate severity has been reported in approximately 5% of patients treated with rosiglitazone during clinical trials. Therefore, caution must be taken when this agent is administered to patients with heart failure. Rosiglitazone has also been associated with elevations of total, LDL, and HDL cholesterol during clinical trials. However, the LDL:HDL cholesterol ratio or the total:HDL cholesterol ratio has mostly been observed to be unchanged. Although liver toxicity has not been observed with rosiglitazone during clinical trials, the safety of this drug for long-term usage and in larger patient populations remains to be established in further clinical studies and in postmarketing experience. PMID- 11728278 TI - Pediatric radiofrequency ablation for tachyarrhythmias: the Westchester Medical Center experience. AB - Radiofrequency ablation, first performed in the late 1980s, has become a standard therapeutic procedure for tachyarrhythmias in children and adults. This procedure can achieve a high degree of success with minimal complications. This report describes the pediatric radiofrequency ablation experience at Westchester Medical Center, including 89 consecutive children who have undergone 104 ablations of 96 arrhythmia substrates with an overall success rate of 95%. The series is analyzed in the context of previously published series of pediatric radiofrequency ablations. Unresolved issues in pediatric ablation related to anesthesia, learning curve, necessity for hospital admission, anticoagulation protocol, and technical aspects of the procedure are discussed. PMID- 11728279 TI - Left atrial compression from severe aortic root dilation and scoliosis. PMID- 11728280 TI - The use of inhaled nitric oxide in newborn medicine. AB - Persistent pulmonary hypertension is common in neonates with respiratory failure. It is characterized by pulmonary hypertension and extrapulmonary right-to-left shunting across the foramen ovale and ductus arteriosus. Nitric oxide has been found to be an important inter- and intracellular messenger in virtually every organ in the body. Inhaled nitric oxide (iNO) is a selective pulmonary vasodilator. The use of iNO improves oxygenation in severely hypoxemic neonates, showing both acute and sustained improvement. It decreases pulmonary arterial pressure, and improves ventilation-perfusion mismatch. This, in turn, may lessen the need for extracorporeal membrane oxygenation. This article reviews the discovery of nitric oxide, its metabolism, its use in persistent pulmonary hypertension of the newborn (PPHN), long-term follow-up, and safety issues related to NO. It has recently been approved by the FDA for the treatment of hypoxic respiratory failure associated with PPHN. PMID- 11728281 TI - Management of hypercholesterolemia in childhood and adolescence. AB - There is strong evidence that the onset of atherosclerosis occurs in childhood. Identifying and treating children and adolescents at risk for hypercholesterolemia should lead to a decrease in adult atherosclerotic disease. Based on current information, and the National Cholesterol Education Program (NCEP) guidelines, screening in children and adolescents should be limited to those individuals with specific cardiac risk factors or those from families with a strong history of atherosclerotic disease. Treatment of identified patients should be initiated with dietary control. Subsequent use of cholesterol-lowering medication is best limited to those patients who fail at least 6 months of dietary control measures. Drug therapy includes the use of bile acid sequestrants, nicotinic acid and, more recently, 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors. There has been limited experience with HMG CoA reductase inhibitors in children and adolescents. However, preliminary data suggests that they are both more effective and have less side effects than either bile acid sequestrants or niacin. Long-term cohort studies will be needed to determine whether screening and treating children and adolescents with hypercholesterolemia is truly of long-term benefit and, if so, which treatment strategies will be preferred. PMID- 11728283 TI - Pulmonary hypertension in infancy and childhood. AB - Abnormalities of pulmonary artery pressure and resistance continue to complicate many varieties of cardiovascular problems in childhood. Much recent effort has been devoted to understanding the cellular mechanisms underlying the pathophysiology of pulmonary hypertension, centering on endothelial cell dysfunction as a principal factor. Defects in the vasodilation machinery of the endothelial cell, such as overexpression of vasoconstrictor elements, have been implicated in various forms of pulmonary hypertension. This includes pulmonary hypertension that is secondary to congenital heart disease, and the primary forms that occur in older children and in neonates. In addition, experimental methods assessing cyclic adenosine monophosphate-mediated and cyclic guanosine monophosphate-mediated vasoreactivity suggest a possible genetic basis in the responses of the pulmonary microvasculature. This article reviews some of the current information that has been developed along these lines, and explores the implications of these data for therapeutic strategies to treat this complex problem. PMID- 11728282 TI - Treatment of heart failure in infants and children. AB - In its simplest and most succinct definition, heart failure can be defined as an inability of the heart to meet the metabolic demands of the body. Despite the diverse etiologies of heart failure in the pediatric population, the presentation of heart failure represents a common constellation of symptoms, signs, and physical findings. In infants, an inability to maintain growth either secondary to decreased nutritional intake or an increased catabolic state is a hallmark of heart failure. Infants exhibit increased sympathetic tone with excessive diaphoresis and increased heart rate. Physical findings in the infants with congestive heart failure (CHF) include increased work of breathing, tachypnea and hepatomegaly. In older children, in contrast, new onset heart failure may be less overtly symptomatic. Malaise, decrease in the level of daily activity, and weight loss may be present. Symptoms of abdominal pain and nausea and anorexia can be present and sometimes divert attention from the real etiology. Physical findings include rales and peripheral edema. If there is hepatomegaly, there will likely be hepatic tenderness as well. A gallop rhythm and tachycardia are commonly present. The long-term treatment of CHF in children includes digoxin, diuretics and afterload reduction with angiotensin-converting enzyme (ACE) inhibitors. Digoxin decreases sympathetic tone and improves growth in infants. Diuretics should be used to relieve symptoms but may not be necessary in all children. ACE inhibitors are increasingly valuable in maintaining cardiac function long term. New uses of medications include the addition of spironalactone (Aldactone, G. D. Searle & Co., Chicago, IL) which, in adults, has been shown to significantly decrease both the death rate from CHF and the need for hospitalization. Beta Blockers have been used in children in limited studies and may have a role in the treatment of patients with idiopathic dilated cardiomyopathy. Surgical treatment, such as partial vectriculectomy, has shown short-term benefit and has been used sparingly in infants. PMID- 11728284 TI - Why lowering blood pressure is not enough: the hypertension syndrome and the clinical context of cardiovascular risk reduction. AB - Although drug treatment for hypertension has been widely used for many years, overall control of this disease continues to be less than optimal. Hypertension treatment has had a positive impact on stroke rates, but rates of coronary heart disease remain high. One reason for this lack of success in treatment is that physicians have focused too narrowly on lowering blood pressure. Blood pressure is a critical element of hypertension, but many other interrelated risk factors contribute to form a complex syndrome of hypertension. Studies show that young adult subjects who do not yet have high blood pressure but do have a family history of hypertension exhibit increased incidence of other risk factors, such as high cholesterol, left ventricular dysfunction, reduced arterial compliance, and insulin resistance. These patients with "normotensive hypertension" share many characteristics with patients with fully developed hypertension, both uncontrolled and controlled. The link between these many risk factors and high blood pressure seems to lie in endothelial cell balance. If this balance is upset, elements of the hypertension syndrome start to become manifest, often before blood pressure becomes elevated. Angiotensin-converting enzyme inhibitors can restore this critical balance, improving overall arterial health and arterial compliance and thus preventing the progression of the hypertension syndrome to high blood pressure and, ultimately, target organ damage. PMID- 11728285 TI - On arterial physiology, pathophysiology of vascular compliance, and cardiovascular disease. AB - Traditionally, the main emphasis in hypertension treatment has been on lowering diastolic blood pressure. Recently, this emphasis has been shifting toward systolic blood pressure and pulse pressure, the latter of which might be a better indicator of future clinical events than either blood pressure reading alone or in combination. Increased pulse pressure indicates increased arterial stiffness and hence is commonly seen in older subjects. As patients age and vessels stiffen, there is a resulting loss of arterial compliance, the ability of the vessel to store blood volume temporarily as it is ejected with each systole. The arterial system acts like a Windkessel, or pump, as it converts intermittent flow from the heart into continuous flow to the organs. The process of stiffening occurs via vascular remodeling, a redistribution of the heterogeneous elements of the vascular wall. Endothelial dysfunction can trigger this remodeling process, increasing stiffness, raising blood pressure and pulse pressure, and ultimately leading to atherosclerosis, plaque formation, and attendant clinical events. Because angiotensin-converting enzyme inhibitors and calcium antagonists can restore arterial compliance, they are suitable choices for hypertension treatment when it is complicated by vascular stiffness. PMID- 11728286 TI - The role of the renin-angiotensin system in vascular health: use of ACE inhibition to improve vascular function. AB - Hypertension is a syndrome in which dysfunction of the blood vessels and of neuroendocrine systems play a major role. Arteriosclerosis and atherosclerosis are distinct processes in arterial disease. Arteriosclerosis begins with thickening and hardening of the arterial wall, with loss of elasticity. Atherosclerosis is an inflammatory process that begins with endothelial dysfunction, and leads to vascular occlusion. The endothelial dysfunction that causes atherosclerosis and coronary artery disease results from an imbalance between nitric oxide and angiotensin-II. This balance becomes tipped toward angiotensin-II in the presence of hypertension. When arteries stiffen as a result of hypertension and aging, they are not able to store blood volume properly, systolic blood pressure rises, and pulse pressure widens. Because the tissue renin-angiotensin system is responsible for promoting the adverse structural arterial changes, inhibition of angiotensin-converting enzyme (ACE) is effective in restoring the health of both large and small arteries by blocking angiotensin and by increasing bradykinin. PMID- 11728287 TI - Increased vascular compliance/decreased cardiovascular risk: what the studies tell us. AB - Increasing arterial stiffness and decreasing arterial compliance are now thought to occur at the beginning of the hypertension disease process. Decreased arterial compliance is associated with isolated systolic blood pressure elevations. Many clinical trials of isolated systolic hypertension provide indirect evidence that improving compliance lowers the risk of cardiovascular disease. The Systolic Hypertension in the Elderly Program showed that antihypertensive therapy in older patients with isolated systolic hypertension lowers systolic blood pressure and narrows pulse pressure without unduly lowering diastolic blood pressure. Treatment over 5 years significantly reduced the incidence of stroke, nonfatal myocardial infarction and coronary death, all cardiovascular events, and all cause mortality. The Systolic Hypertension in Europe and Systolic Hypertension in China trials showed similar effects on pulse pressure and on clinical end points. The Heart Outcomes Prevention Evaluation was a primary prevention study of the effect of an angiotensin-converting enzyme (ACE) inhibitor in subjects who were not necessarily hypertensive but were at risk for cardiovascular events. With minimal lowering of blood pressure, ramipril therapy provided significant risk reduction in all major end points--overall mortality, stroke, myocardial infarction, and cardiovascular death. ACE inhibitors restore endothelial cell balance to improve arterial compliance, thus they can provide benefits beyond lowering blood pressure. PMID- 11728288 TI - The ABC Trial. PMID- 11728289 TI - Evaluation of candesartan cilexetil in black patients with systemic hypertension: the ABC Trial. AB - To assess the efficacy and tolerability of the angiotensin-II receptor blocker candesartan cilexetil, the Association of Black Cardiologists (ABC) coordinated a 12-week, multicenter, double-blind, placebo-controlled clinical trial in 304 black adults with a sitting diastolic blood pressure (DBP) of 91 to 105 mmHg, inclusive. The goal of the ABC Trial was to address previous reports suggesting that drugs that block the renin-angiotensin-aldosterone system may not be effective in controlling BP in hypertensive black individuals. Patients were randomized to receive candesartan cilexetil 16 mg (n = 156) or placebo (n = 148), once daily after a 4- to 5-week single-blind, placebo run-in period. Candesartan cilexetil and placebo doses were doubled if the trough (24 +/- 3 hours after dose) sitting DBP was 90 mmHg or higher after 4 weeks. If the DBP was 90 mmHg or higher at week 8, hydrochlorothiazide 12.5 mg was added to both placebo or candesartan cilexetil 32 mg. Candesartan cilexetil was significantly more effective than placebo in reducing trough sitting systolic BP (SBP) and DBP at week 8 (6.4/5.1 mmHg versus 1.3/2.7 mmHg) and at week 12 (9.3/7.5 mmHg versus 5.7/5.2 mmHg). Hydrochlorothiazide was added in 27 and 50% of patients in the candesartan cilexetil and placebo groups, respectively. Control and responder rates were consistently higher in the candesartan cilexetil treatment group at weeks 8 and 12. Discontinuation and adverse event rates were similar in the two groups. Candesartan cilexetil once daily was effective in reducing BP in a substantial proportion of an exclusively black hypertensive population. The combination of candesartan cilexetil plus hydrochlorothiazide demonstrated additional efficacy while maintaining an excellent tolerability profile. PMID- 11728291 TI - Is it descending thoracic aortic dissection? AB - Complex thoracoabdominal aortic aneurysm repairs are preferably done by a staged approach, using the elephant trunk procedure. Postoperatively, such patients can present as a diagnostic dilemma if subjected to radiologic diagnostic studies such as computerized tomographic (CT) scanning or magnetic resonance imaging (MRI) of the thoracic cavity. The authors report a patient who presented with symptoms of chest pain and underwent CT scanning and subsequently MRI of the chest. Both scans were interpreted to be consistent with aortic dissection; however, the elephant trunk prosthesis was responsible for simulating a dissection. PMID- 11728292 TI - Gene transfer in the cardiovascular system: update 2000. AB - It has been slightly more than 10 years since the first proof-of-concept studies were performed, which demonstrated the feasibility of gene transfer into the heart and vasculature of experimental animals. Since that time there has been a dramatic increase in the nature and sophistication of gene transfer techniques and also in the number of cardiovascular diseases that are potential targets for gene-based therapies. In this article, the authors review the current strategies for gene delivery, including viral and nonviral approaches. The authors also highlight several biologic processes within the cardiovascular system, including restenosis, experimental angiogenesis, heart failure, and atherosclerosis conditions for which gene therapy shows promise. It is hoped that this will provide an update of this therapeutic strategy for the year 2000. PMID- 11728290 TI - Effects of candesartan cilexetil on health-related quality of life in black patients with systemic hypertension in the ABC Trial. AB - Tolerability is an important consideration in evaluating a new antihypertensive agent. This can be assessed informally by conventional patient interviews or more formally with the use of validated health-related quality of life (HRQL) measures assessing the patient's perception of the agent's tolerability. HRQL was a secondary end point of a 12-week, multicenter, double-blind, randomized, placebo controlled study of the efficacy and tolerability of candesartan cilexetil in black patients with systemic hypertension. HRQL was evaluated using the generic Medical Outcomes Study 36-Item Short Form (SF-36) and population- and condition specific Vital Signs Quality of Life Questionnaire (VSQLQ). Data were gathered via face-to-face interviews at screening, baseline, and weeks 8 and 12. Of the 304 patients randomized, 268 were evaluable for the HRQL analysis. Clinical results, reported in the companion article, found that candesartan cilexetil initiated at 16 mg once daily and titrated to 32 mg once daily as needed, with the subsequent addition of hydrochlorothiazide 12.5 mg as needed, was effective for lowering diastolic and systolic blood pressure and was well tolerated based on office interviews. Analyses of patients' perceptions of tolerability found that HRQL was maintained during the 12-week study period, with no significant differences between treatment and placebo groups at the end of double-blind treatment. These results indicate that the HRQL of black patients with systemic hypertension is maintained during treatment with candesartan cilexetil. PMID- 11728293 TI - Primary pulmonary hypertension. AB - Primary pulmonary hypertension (PPH) is a condition characterized by sustained elevation of pulmonary artery pressure (PAP) without demonstrable cause. The most common symptom at presentation is dyspnea. Other complaints include fatigue, chest pain, syncope, leg edema, and palpitations. Right heart catheterization is diagnostic, showing a mean PAP >25 mmHg at rest and >30 mmHg during exercise, with a normal pulmonary capillary wedge pressure. In the National Institutes of Health-PPH registry, the median survival period was 2.8 years. Treatment is aimed at lowering PAP, increasing cardiac output, and decreasing in situ thrombosis. Vasodilators have been used with some success in the treatment of PPH. They include prostacyclin, calcium-channel blockers, nitric oxide and adenosine. Anticoagulation has also been advised for the prevention of deep vein thrombosis, pulmonary embolism, and in situ thromboses of the lungs. New drug treatments under investigation include L-arginine, plasma endothelin-I, and bosentan. Use of oxygen, digoxin, and diuretics for symptomatic relief have also been recommended. Patients with severe PPH refractory to medical management should be considered for surgery. PMID- 11728294 TI - The management of cor pulmonale. AB - Pulmonary artery hypertension occurs when there is a sustained elevation of the mean pulmonary pressure above normal physiologic values. This may then lead to cor pulmonale or enlargement of the right ventricle due to any lung disease in the absence of left heart failure. When the ability of the right ventricle to compensate is overwhelmed, right-sided heart failure can occur. Pulmonary artery hypertension may be idiopathic (primary) or secondary to a variety of lung parenchymal diseases, airways disease, pulmonary circulatory disorders, systemic illnesses, or thoracic mechanical abnormalities. Treatment strategies for cor pulmonale include supplemental oxygen, assisted mechanical ventilation, digoxin, and diuretics. Pulmonary vasodilator compounds should be used with caution because they can compromise gas exchange in cor pulmonale from secondary pulmonary hypertension. Trials with digoxin and inotropic agents have been evaluated. Angiotensin-converting enzyme inhibitors have not shown significant utility, at least acutely. Anticoagulation may decrease mortality in some patients with pulmonary artery hypertension and cor pulmonale. PMID- 11728295 TI - Treatment of venous thromboembolism. AB - Venous thromboembolism (VTE) is a disease entity composed of pulmonary embolism (PE) and deep vein thrombosis (DVT). Anticoagulation, initiated as soon as the diagnosis is suspected, is the treatment of choice. Traditionally, anticoagulation is started with intravenous heparin, and changed to warfarin for long-term treatment. The introduction of unmonitored, subcutaneously administered, low molecular weight heparin has resulted in shorter hospitalizations, reduced the incidence of major bleeding as a complication, and shifted the treatment of VTE for selected patients to the outpatient setting. Thrombolytic therapy has been recommended for patients with life-threatening PE. Technologic advances in catheter embolectomy and fragmentation permit clot resolution in patients in whom thrombolytic therapy is contraindicated. Inferior cava filters can be placed percutaneously in patients at high risk for VTE or those in whom anticoagulation is contraindicated. Because VTE is often clinically silent, prevention of VTE is therefore the most effective means to reduce associated morbidity and mortality. Strategies to prevent VTE have been studied and validated for specific clinical circumstances. PMID- 11728296 TI - Perindopril. AB - Perindopril is the newest angiotensin-converting enzyme inhibitor to be approved in the United States. It is indicated for the treatment of essential hypertension either alone or in combination with other antihypertensives, especially thiazide diuretics. Perindopril must be converted to perindoprilat, its active metabolite. The drug's prolonged duration of action allows for once daily administration. This article reviews clinical trials performed using perindopril alone and in combination to treat essential hypertension. The use of perindopril for congestive heart failure is reviewed. The influence of perindopril on arterial wall properties is discussed. Tolerability, drug interactions, and dosage and administration of the drug are reviewed. PMID- 11728297 TI - An evidence-based approach to acute respiratory distress syndrome. AB - We provide an evidence-based approach to managing patients with acute lung injury and acute respiratory distress syndrome (ARDS). We searched MEDLINE and the Cumulative Index to Nursing and Allied Health for randomized trials evaluating lung-protective ventilation strategies, inhaled nitric oxide, prone positioning, and late-phase corticosteroids for managing these patients, and for additional literature related to long-term follow-up of ARDS survivors. The results of our review suggest that pressure- and volume-limited ventilation, according to the ARDS Network protocol, can reduce mortality for patients with acute lung injury, and so may an "open lung" approach to mechanical ventilation. Those 2 strategies are currently being compared in 2 multicenter randomized trials. Although both inhaled nitric oxide therapy and prone positioning can produce dramatic acute improvements in oxygenation for some patients, there is no evidence that these interventions can benefit patients with respect to patient-important outcomes. Therefore it is unreasonable to be dogmatic about the role of inhaled nitric oxide and prone positioning in ARDS. The role of corticosteroids in the late phase of ARDS is unclear and remains a very important unanswered question. With respect to long-term follow-up, we found that pulmonary dysfunction is probably not a major source of morbidity for ARDS survivors, whereas neuropsychological dysfunction is prominent. Ongoing research may suggest interventions to improve the outcome of ARDS and of critical illness in general. PMID- 11728298 TI - Evidence-based treatments for acute asthma. AB - Asthma is an important health care problem; over 12 million people in the United States suffer from asthma, the majority of whom are young patients. Presentations of acute asthma to emergency departments are common. In the United States, acute asthma presentations account for close to 2 million emergency department visits annually, and these patients often exhibit acute and chronic markers of severe asthma; so controlling asthma is important from many perspectives. We review the evidence for various acute asthma therapies, using the highest levels of evidence, employing systematic reviews (especially those from the Cochrane Collaboration) and evidence from randomized controlled trials to guide therapy decisions. beta agonists and systemic corticosteroids are the cornerstones of initial treatment. Delivery of beta agonists via nebulizer or metered-dose inhaler with spacer device appear to be similarly efficacious. However, recent evidence from studies involving children and adults indicate that addition of ipratropium bromide to early beta agonist treatments may reduce airway obstruction and reduce hospital admissions, especially for more severe asthma. Evidence from systematic reviews indicates that intravenous magnesium sulfate may provide similar benefits in severe asthma. Antibiotics, intravenous beta agonists, and intravenous aminophylline have been shown to add little and may increase adverse effects. Treatment for discharged patients should include systemic corticosteroids for 5-7 days, for all but the mildest asthma. Addition of inhaled corticosteroids should be considered for most patients, since evidence suggests that inhaled corticosteroids may reduce relapses and improve quality of life. Alternative treatments such as long-acting beta agonists and leukotriene antagonists remain unproven in this setting. Linking a discharge plan to close follow-up and asthma education (especially an action plan) needs to be encouraged. Acute asthma is a common problem and treatment has improved dramatically over the past 10 years. Employing the evidence-based practice outlined above should reduce the burden of acute asthma on patients and the health care system. PMID- 11728299 TI - Evidenced-based airway management. AB - The increased polarity of evidenced-based medicine has changed the approach to evaluation of medical interventions and treatments, either placing them on firm scientific foundations or suggesting that evidence is inadequate to strongly support current or innovative practices. Airway management devices and techniques are essential and common components of clinical care. It is the purpose of this review to identify the levels of evidence that support common and novel techniques in airway management. Very few airway management techniques are supported by large, prospective randomized trials. In resuscitation, defibrillation should take priority over airway management. This is supported by animal studies and several reported series of patient experiences and will never be subjected to a prospective randomized trial. Substantial evidence supports the use of noninvasive ventilation, subglottic endotracheal tube secretion removal, changing ventilator circuits no more frequently than every 7 days, and the use of selective digestive decontamination with systemic antibiotics to reduce the incidence of ventilator-associated pneumonia. Little evidence supports using other measures such as elevating the head of the bed to 30% (but this costs nothing and is intrinsically attractive), use of heat and moisture exchangers, kinetic bed therapy, early tracheotomy, or lung secretion removal techniques to reduce ventilator-associated pneumonia. Percutaneous tracheotomy currently can only be recommended over open surgical tracheotomy based on cost, convenience and late stomal complications; it may be associated with a slightly higher morbidity and mortality. PMID- 11728300 TI - Weaning from mechanical ventilation: the evidence from clinical research. AB - Mechanical ventilation incurs substantial morbidity, mortality, and costs. Both premature extubation and delayed extubation can cause harm. Therefore, weaning that is both expeditious and safe is highly desirable. The purpose of this review is to summarize the literature related to weaning modes, spontaneous breathing trials, weaning predictors, weaning with noninvasive positive pressure ventilation, and weaning protocols. We used 5 computerized databases and a duplicate independent review process to select articles for this review. We included randomized clinical trials evaluating any weaning interventions and nonrandomized trials of weaning predictors, with a focus on studies reporting clinically important outcomes. We abstracted quantitative data using several metrics and pooled results across studies only when our assessment of the patients, interventions, and outcomes indicated that pooling was legitimate. The available clinical research evidence suggests that, for progressive weaning of the level of mechanical support, it may be best to choose modes other than synchronized intermittent mandatory ventilation and it is unreasonable to be dogmatic about the use of other modes. There may also be substantial benefits to early extubation with back-up institution of noninvasive positive pressure ventilation, as needed, though this remains an experimental approach. For trials of spontaneous breathing, low levels of pressure support may hasten extubation. We did not uncover any consistently powerful weaning predictors, suggesting that formal use of predictors in patients being considered for reduction or discontinuation of mechanical support is unlikely to improve patient care. The likely explanation is that clinicians already fully consider information from weaning predictors in choosing patients for trials of reduction or discontinuation of mechanical ventilation. Finally, implementation of respiratory therapist- or nurse-driven protocols may be useful for all phases of weaning, and clinicians should adopt daily assessment for a trial of unassisted breathing as a safe method to reduce the duration of mechanical ventilation. PMID- 11728301 TI - Evidence-based medicine and sleep apnea. AB - There is an increasing body of evidence that obstructive sleep apnea (OSA) affects systemic blood pressure and quality of life. The evidence linking OSA with other types of cardiovascular morbidity and mortality is not as strong. We review the current literature on the health impact, diagnosis, and treatment of OSA and provide a framework for evaluating levels of evidence in each of these areas, with the focus on high quality studies. Clinical prediction rules and unattended portable monitoring devices, such as digital oximeters, are inexpensive and have clinically useful likelihood ratios, sensitivity, and specificity, so they can be considered clinically useful in the evaluation of individuals suspected of having OSA. This approach has the potential to change the probability of disease in some patients to a level at which polysomnography is unnecessary. Recent randomized, placebo-controlled trials have indicated that treating OSA with continuous positive airway pressure improves quality of life, driving simulator performance, blood pressure, and sleepiness. The evidence in support of mandibular advancement is less strong, and surgical measures should be viewed as an option only after continuous positive airway pressure and mandibular advancement have been considered, given the paucity of evidence to support surgery. The quality of research done on the prognosis, diagnosis, and treatment of OSA has improved substantially in recent years. However, there remains a need for additional studies that evaluate ambulatory monitors and for larger randomized controlled trials of therapy with longer patient follow-up to verify the range of benefits that OSA patients can expect from the various treatment modalities. PMID- 11728302 TI - The limits of evidence-based medicine. AB - The importance of clinical research for the practice of clinical medicine is immense and undeniable. Yet the type of knowledge gained from clinical research, referred to here as "empirical evidence," is itself insufficient to provide for optimal clinical care. A gap exists between empirical evidence and clinical practice. Proponents of evidence-based medicine have clearly acknowledged one aspect of this gap: the part that requires the consideration of values, both patient and professional, prior to arriving at medical decisions. Not as clearly recognized, however, is the gap that exists due to the fact that empirical evidence is not directly applicable to individual patients, as the knowledge gained from clinical research does not directly answer the primary clinical question of what is best for the patient at hand. Proponents of evidence-based medicine have made a conceptual error by grouping knowledge derived from clinical experience and physiologic rationale under the heading of "evidence" and then have compounded the error by developing hierarchies of "evidence" that relegate these forms of medical knowledge to the lowest rungs. Empirical evidence, when it exists, is viewed as the "best" evidence on which to make a clinical decision, superseding clinical experience and physiologic rationale. But these latter forms of medical knowledge differ in kind, not degree, from empirical evidence and do not belong on a graded hierarchy. As they differ in kind, these other forms of medical knowledge can be viewed as complementary to empirical evidence and their incorporation necessary to overcome the intrinsic gap noted above. Clinicians, then, need to incorporate knowledge from 5 distinct areas into each medical decision: (1) empirical evidence, (2) experiential evidence, (3) physiologic principles, (4) patient and professional values, and (5) system features. The relative weight given to each of these areas is not predetermined, but varies from case to case. PMID- 11728303 TI - Understanding why we agree on the evidence but disagree on the medicine. AB - Despite the fact that physicians, nurses, and respiratory therapists share a common training in scientific evidence and frequently share a common understanding of the evidence, debates occur about the proper management of individual patients. Clinicians may interpret this as a failure of evidence-based medicine, when, in fact, evidence is only one of many factors that play a role in clinical decision-making. By clearly understanding the other factors that affect clinical decision making-prior beliefs about treatment effectiveness, patient and clinician values, and clinical expertise-clinicians can better understand which clinical disagreements are about scientific evidence and which are about other factors. PMID- 11728304 TI - Role of scaffolds in MAP kinase pathway specificity revealed by custom design of pathway-dedicated signaling proteins. AB - BACKGROUND: Signal transduction pathways with shared components must be insulated from each other to avoid the inappropriate activation of multiple pathways by a single stimulus. Scaffold proteins are thought to contribute to this specificity by binding select substrates. RESULTS: We have studied the ability of scaffold proteins to influence signaling by the yeast kinase Ste11, a MAPKKK molecule that participates in three distinct MAP kinase pathways: mating, filamentation, and HOG. We used protein fusions to force Ste11 to associate preferentially with a subset of its possible binding partners in vivo, including Ste5, Ste7, and Pbs2. Signaling became confined to a particular pathway when Ste11 was covalently attached to these scaffolds or substrates. This pathway bias was conferred upon both stimulus-activated and constitutively active forms of Ste11. We also used membrane-targeted derivatives of the mating pathway scaffold, Ste5, to show that stimulus-independent signaling initiated by this scaffold remained pathway specific. Finally, we demonstrate that loss of pathway insulation has a negative physiological consequence, as nonspecific activation of both the HOG and mating pathways interfered with proper execution of the mating pathway. CONCLUSIONS: The signaling properties of these kinase fusions support a model in which scaffold proteins dictate substrate choice and promote pathway specificity by presenting preferred substrates in high local concentration. Furthermore, insulation is inherent to scaffold-mediated signaling and does not require that signaling be initiated by pathway-specific stimuli or activator proteins. Our results give insight into the mechanisms and physiological importance of pathway insulation and provide a foundation for the design of customized signaling proteins. PMID- 11728305 TI - Separase is required for chromosome segregation during meiosis I in Caenorhabditis elegans. AB - BACKGROUND: Chromosome segregation during mitosis and meiosis is triggered by dissolution of sister chromatid cohesion, which is mediated by the cohesin complex. Mitotic sister chromatid disjunction requires that cohesion be lost along the entire length of chromosomes, whereas homolog segregation at meiosis I only requires loss of cohesion along chromosome arms. During animal cell mitosis, cohesin is lost in two steps. A nonproteolytic mechanism removes cohesin along chromosome arms during prophase, while the proteolytic cleavage of cohesin's Scc1 subunit by separase removes centromeric cohesin at anaphase. In Saccharomyces cerevisiae and Caenorhabditis elegans, meiotic sister chromatid cohesion is mediated by Rec8, a meiosis-specific variant of cohesin's Scc1 subunit. Homolog segregation in S. cerevisiae is triggered by separase-mediated cleavage of Rec8 along chromosome arms. In principle, chiasmata could be resolved proteolytically by separase or nonproteolytically using a mechanism similar to the mitotic "prophase pathway." RESULTS: Inactivation of separase in C. elegans has little or no effect on homolog alignment on the meiosis I spindle but prevents their timely disjunction. It also interferes with chromatid separation during subsequent embryonic mitotic divisions but does not directly affect cytokinesis. Surprisingly, separase inactivation also causes osmosensitive embryos, possibly due to a defect in the extraembryonic structures, referred to as the "eggshell." CONCLUSIONS: Separase is essential for homologous chromosome disjunction during meiosis I. Proteolytic cleavage, presumably of Rec8, might be a common trigger for the first meiotic division in eukaryotic cells. Cleavage of proteins other than REC-8 might be necessary to render the eggshell impermeable to solutes. PMID- 11728306 TI - Coordination of cell polarization and migration by the Rho family GTPases requires Src tyrosine kinase activity. AB - BACKGROUND: The ability of a cell to polarize and move is governed by remodeling of the cellular adhesion/cytoskeletal network that is in turn controlled by the Rho family of small GTPases. However, it is not known what signals lie downstream of Rac1 and Cdc42 during peripheral actin and adhesion remodeling that is required for directional migration. RESULTS: We show here that individual members of the Rho family, RhoA, Rac1, and Cdc42, direct the specific intracellular targeting of c-Src tyrosine kinase to focal adhesions, lamellipodia, or filopodia, respectively, and that the adaptor function of c-Src (the combined SH3/SH2 domains coupled to green fluorescent protein) is sufficient for targeting. Furthermore, Src's catalytic activity is absolutely required at these peripheral cell-matrix attachment sites for remodeling that converts RhoA dependent focal adhesions into smaller focal complexes along Rac1-induced lamellipodia (or Cdc42-induced filopodia). Consequently, cells in which kinase deficient c-Src occupies peripheral adhesion sites exhibit impaired polarization toward migratory stimuli and reduced motility. Furthermore, phosphorylation of FAK, an Src adhesion substrate, is suppressed under these conditions. CONCLUSIONS: Our findings demonstrate that individual Rho GTPases specify Src's exact peripheral localization and that Rac1- and Cdc42-induced adhesion remodeling and directed cell migration require Src activity at peripheral adhesion sites. PMID- 11728307 TI - An epithelial cell destined for apoptosis signals its neighbors to extrude it by an actin- and myosin-dependent mechanism. AB - BACKGROUND: Simple epithelia encase developing embryos and organs. Although these epithelia consist of only one or two layers of cells, they must provide tight barriers for the tissues that they envelop. Apoptosis occurring within these simple epithelia could compromise this barrier. How, then, does an epithelium remove apoptotic cells without disrupting its function as a barrier? RESULTS: We show that apoptotic cells are extruded from a simple epithelium by the concerted contraction of their neighbors. A ring of actin and myosin forms both within the apoptotic cell and in the cells surrounding it, and contraction of the ring formed in the live neighbors is required for apoptotic cell extrusion, as injection of a Rho GTPase inhibitor into these cells completely blocks extrusion. Addition of apoptotic MDCK cells to an intact monolayer induces the formation of actin cables in the cells contacted, suggesting that the signal to form the cable comes from the dying cell. The signal is produced very early in the apoptotic process, before procaspase activation, cell shrinkage, or phosphatidylserine exposure. Remarkably, electrical resistance studies show that epithelial barrier function is maintained, even when large numbers of dying cells are being extruded. CONCLUSIONS: We propose that apoptotic cell extrusion is important for the preservation of epithelial barrier function during cell death. Our results suggest that an early signal from the dying cell activates Rho in live neighbors to extrude the apoptotic cell out of the epithelium. PMID- 11728308 TI - Direct imaging of in vivo neuronal migration in the developing cerebellum. AB - The upper rhombic lip (URL), a germinal zone in the dorsoanterior hindbrain, has long been known to be a source for neurons of the vertebrate cerebellum. It was thought to give rise to dorsally migrating granule cell precursors (Figure 1e); however, recent fate mapping studies have questioned the exclusive contributions of the URL to granule cells. By taking advantage of the clarity of the zebrafish embryo during the stages of brain morphogenesis, we have followed the fate of neuronal precursor cells generated within the upper rhombic lip directly. Combining a novel GFP labeling strategy with in vivo time-lapse imaging, we find, contrary to the former view, that most URL-descendants migrate anterior toward the midhindbrain boundary (MHB) and then course ventrally along the MHB (Figure 1f). As the migrating neuronal precursors reach the MHB, they form ventrally extending projections, likely axons, and continue ventral migration to settle outside of the cerebellum, in the region of the ventral brainstem. Thus, we define a new pathway for URL-derived neuronal precursor cells consistent with the recent fate maps. In addition, our results strongly suggest that the MHB plays a crucial role, not only in induction and patterning of the cerebellar anlage, but also in organizing its later morphogenesis by influencing cell migration. PMID- 11728309 TI - Nr-CAM and neurofascin interactions regulate ankyrin G and sodium channel clustering at the node of Ranvier. AB - Voltage-dependent sodium (Na(+)) channels are highly concentrated at nodes of Ranvier in myelinated axons and play a key role in promoting rapid and efficient conduction of action potentials by saltatory conduction. The molecular mechanisms that direct their localization to the node are not well understood but are believed to involve contact-dependent signals from myelinating Schwann cells and interactions of Na(+) channels with the cytoskeletal protein, ankyrin G. Two cell adhesion molecules (CAMs) expressed at the axon surface, Nr-CAM and neurofascin, are also linked to ankyrin G and accumulate at early stages of node formation, suggesting that they mediate contact-dependent Schwann cell signals to initiate node development. To examine the potential role of Nr-CAM in this process, we treated myelinating cocultures of DRG (dorsal root ganglion) neurons and Schwann cells with an Nr-CAM-Fc (Nr-Fc) fusion protein. Nr-Fc had no effect on initial axon-Schwann cell interactions, including Schwann cell proliferation, or on the extent of myelination, but it strikingly and specifically inhibited Na(+) channel and ankyrin G accumulation at the node. Nr-Fc bound directly to neurons and clustered and coprecipitated neurofascin expressed on axons. These results provide the first evidence that neurofascin plays a major role in the formation of nodes, possibly via interactions with Nr-CAM. PMID- 11728310 TI - Apoptotic mimicry by an obligate intracellular parasite downregulates macrophage microbicidal activity. AB - Programmed cell death by apoptosis of unnecessary or potentially harmful cells is clearly beneficial to multicellular organisms. Proper functioning of such a program demands that the removal of dying cells proceed without an inflammatory reaction. Phosphatidylserine (PS) is one of the ligands displayed by apoptotic cells that participates in their noninflammatory removal when recognized by neighboring phagocytes. PS ligation induces the release of transforming growth factor-beta (TGF-beta), an antiinflammatory cytokine that mediates the suppression of macrophage-mediated inflammation. In Hydra vulgaris, an organism that stands at the base of metazoan evolution, the selective advantage provided by apoptosis lies in the fact that Hydra can survive recycling apoptotic cells by phagocytosis. In unicellular organisms, it has been proposed that altruistic death benefits clonal populations of yeasts and trypanosomatids. Now we show that advantageous features of the apoptotic process can operate without death as the necessary outcome. Leishmania spp are able to evade the killing activity of phagocytes and establish themselves as obligate intracellular parasites. Amastigotes, responsible for disease propagation, similar to apoptotic cells, inhibit macrophage activity by exposing PS. Exposed PS participates in amastigote internalization. Recognition of this moiety by macrophages induces TGF-beta secretion and IL-10 synthesis, inhibits NO production, and increases susceptibility to intracellular leishmanial growth. PMID- 11728311 TI - C. elegans Rb, NuRD, and Ras regulate lin-39-mediated cell fusion during vulval fate specification. AB - The tumor suppressor Rb and the NuRD (nucleosome remodeling and histone deacetylation) complex have been implicated in transcriptional repression during cell cycle progression and cell fate specification. The Rb/E2F complex physically interacts with and thus recruits the NuRD complex to actively repress transcription. Caenorhabditis elegans counterparts of Rb, E2F/DP, and some NuRD complex components appear to function in a common class B synthetic Multivulva (synMuv) pathway to antagonize RTK/Ras signaling during vulval fate specification. Therefore, it has been suggested that they function together in a single complex to repress vulva-specific gene transcription. However, little is known about the in vivo interactions between these class B synMuv genes and their relationships with other pathways in specific cellular processes during vulval development. We show that C. elegans Rb/E2F and NuRD complexes antagonize Ras activity by controlling a lin-39 Hox-mediated cell fusion event that regulates the competence of vulval cells. Interestingly, Rb/E2F and NuRD complexes exhibit very different genetic properties. While the NuRD complex negatively regulates lin-39 Hox activity, likely by downregulating its expression, RB/E2F appears to play dual roles in regulating lin-39: a negative role in controlling its activity and a previously uncharacterized positive role in regulating its expression. PMID- 11728312 TI - Regulating c-Ras function. cholesterol depletion affects caveolin association, GTP loading, and signaling. AB - Cholesterol-rich and caveolin-containing microdomains of the plasma membrane, termed "caveolae," have been implicated in signal transduction. However, the role of caveolae in regulating the Ras-MAP kinase cascade is incompletely understood. The mammalian Ras isoforms (H, N, and K) use different membrane anchors to attach to the plasma membrane and thereby may localize to functionally distinct microdomains, which might explain isoform-specific signaling. Here, we show that, in Cos epithelial cells, endogenous K-Ras colocalizes largely with caveolin, whereas N-Ras localizes to both caveolar and noncaveolar subdomains; H-Ras localization was below detection limits. We find that epidermal growth factor (EGF) activates N-Ras but fails to activate K-Ras in these cells. Extraction of cholesterol with methyl-beta-cyclodextrin disrupts complex formation between caveolin and K- and N-Ras and, strikingly, enables EGF to activate both K-Ras and N-Ras. While cholesterol depletion enhances GTP-loading on total c-Ras, activation of the downstream MEK-MAP kinase cascade by EGF and lysophosphatidic acid but not that by phorbol ester is inhibited. Thus, plasma membrane cholesterol is essential for negative regulation of c-Ras isoforms (complexed to caveolin), as well as for mitogenic signaling downstream of receptor-activated c Ras. PMID- 11728313 TI - A yeast homolog of chromatin assembly factor 1 is involved in early ribosome assembly. AB - Cells have a recurrent need for the correct assembly of protein-nucleic acid complexes. We have studied a yeast homolog of the smallest subunit of chromatin assembly factor 1 (CAF1), encoded by YMR131c and termed "RRB1". Unlike other yeast homologs, Msi1p, and Hat2p, Rrb1p is essential for cell viability. Impairment of Rrb1p function results in decreased levels of free 60S ribosomal subunits and the appearance of half-mer polysomes, suggesting its involvement in ribosome biogenesis. Using tandem affinity purification (TAP ) combined with mass spectrometry, we show that Rrb1p is associated with ribosomal protein L3. A fraction of Rrb1p is also found in a protein-precursor rRNA complex containing at least ten other early-assembling ribosomal proteins. We propose that Rrb1p is required for proper assembly of preribosomal particles during early ribosome biogenesis, presumably by targeting L3 onto the 35S precursor rRNA. This action may resemble the mechanism by which CAF1 assembles histones H3/H4 onto newly replicated DNA. PMID- 11728314 TI - CPR5 is involved in cell proliferation and cell death control and encodes a novel transmembrane protein. AB - Plants often respond to pathogens by sacrificing cells at the infection site. This type of programmed cell death is mimicked by the constitutive pathogene response5 (cpr5) mutant in Arabidopsis in the absence of pathogens, suggesting a role for CPR5 in programmed cell death control. The analysis of the cellular phenotypes of two T-DNA-tagged cpr5 alleles revealed an additional role for CPR5 in the regulation of endoreduplication and cell division. In cpr5 mutant trichomes, endoreduplication cycles stop after two rounds instead of four, and trichome cells have fewer branches than normal. Eventually, cpr5 trichomes die, the nucleus disintegrates, and the cell collapses. Similarly, leaf growth stops earlier than in wild-type, and, frequently, regions displaying spontaneous cell death are observed. The cloning of the CPR5 gene revealed a novel putative transmembrane protein with a cytosolic domain containing a nuclear-targeting sequence. The dual role of CPR5 in cell proliferation and cell death control suggests a regulatory link between these two processes. PMID- 11728315 TI - Grasping the past. delay can improve visuomotor performance. AB - "Optic ataxia" is caused by damage to the human posterior parietal cortex (PPC). It disrupts all components of a visually guided prehension movement, not only the transport of the hand toward an object's location, but also the in-flight finger movements pretailored to the metric properties of the object. Like previous cases, our patient (I.G.) was quite unable to open her handgrip appropriately when directly reaching out to pick up objects of different sizes. When first tested, she failed to do this even when she had previewed the target object 5 s earlier. Yet despite this deficit in "real" grasping, we found, counterintuitively, that I.G. showed good grip scaling when "pantomiming" a grasp for an object seen earlier but no longer present. We then found that, after practice, I.G. became able to scale her handgrip when grasping a real target object that she had previewed earlier. By interposing catch trials in which a different object was covertly substituted for the original object during the delay between preview and grasp, we found that I.G. was now using memorized visual information to calibrate her real grasping movements. These results provide new evidence that "off-line" visuomotor guidance can be provided by networks independent of the PPC. PMID- 11728316 TI - Fresh look at potential cannabis therapies. PMID- 11728317 TI - Yersinia pestis: sequence sheds light on the plague's past. PMID- 11728319 TI - Ethylene. PMID- 11728320 TI - DNA replication checkpoint. PMID- 11728321 TI - Genome instability: McClintock revisited. AB - Recent studies in yeast have shed light on the molecular mechanisms by which telomere dysfunction leads to chromosome fusions. Furthermore, examination of the consequences of telomerase loss in mice suggests that only a few critically short telomeres may be sufficient to promote genomic instability. PMID- 11728322 TI - mRNA export: travelling with DEAD box proteins. AB - Recent studies have shown that the putative RNA helicase protein UAP56 and its yeast homologue Sub2p are not only involved in pre-mRNA splicing but also required for the export of mRNA out of the nucleus, even if the mRNA is encoded by an intron-less gene. PMID- 11728323 TI - Cognitive neuroscience: forgetting of things past. AB - Recent functional imaging and electrophysiological results indicate that failure to remember experiences can result from a decreased recruitment of encoding processes that build effective memories and an increased recruitment of alternative mechanisms that may impair effective learning. PMID- 11728324 TI - ABA signalling: a messenger's FIERY fate. AB - There is considerable circumstantial evidence that the Ca(2+)-mobilizing second messenger IP(3) is involved in plant responses to the drought hormone abscisic acid. More direct evidence for this has now come from studies in which endogenous IP(3) levels have been manipulated in plants. PMID- 11728325 TI - Membrane repair: Ca(2+)-elicited lysosomal exocytosis. AB - Cells in exposed positions are subject to injury and therefore need membrane repair mechanisms. Ca(2+) entry inevitably follows membrane rupture and recent studies indicate that this elicits repair via Ca(2+)-activated exocytosis of lysosomes, regulated by lysosomal synaptotagmin VII. PMID- 11728326 TI - Cell polarity: no need to reinvent the wheel. AB - Epithelial cells are polarized along their apical-basal axis and in some cases also within the plane of the epithelium, a phenomenon called planar polarity. Recent studies have now shown that these two types of polarity are controlled by a common set of genes. PMID- 11728327 TI - DNA replication: stable driving prevents fatal smashes. AB - Cells respond to DNA damage during S phase by slowing chromosome replication. Recent results have shed light on the mechanism by which this 'intra-S phase' checkpoint is implemented. PMID- 11728328 TI - Root patterning: SHORT ROOT on the move. AB - The radial pattern of the plant root is determined by the action of two transcription factors, SHORT ROOT and SCARECROW, which are produced in different cell types. The SHORT ROOT protein has now been shown to move from cell to cell, regulating transcription of the downstream SCARECROW gene in the target cells. PMID- 11728329 TI - Central pattern generators and the control of rhythmic movements. AB - Central pattern generators are neuronal circuits that when activated can produce rhythmic motor patterns such as walking, breathing, flying, and swimming in the absence of sensory or descending inputs that carry specific timing information. General principles of the organization of these circuits and their control by higher brain centers have come from the study of smaller circuits found in invertebrates. Recent work on vertebrates highlights the importance of neuro modulatory control pathways in enabling spinal cord and brain stem circuits to generate meaningful motor patterns. Because rhythmic motor patterns are easily quantified and studied, central pattern generators will provide important testing grounds for understanding the effects of numerous genetic mutations on behavior. Moreover, further understanding of the modulation of spinal cord circuitry used in rhythmic behaviors should facilitate the development of new treatments to enhance recovery after spinal cord damage. PMID- 11728330 TI - FERMing up the synapse. AB - The immunological synapse is characterized by the reorganization of membrane proteins at the immunological synapse. Active cytoskeletal mechanisms are involved in recruiting the TCR, accessory molecules, and the integrin LFA-1 to the contact area. Other molecules like CD43 are excluded from the contact area, but the mechanism of exclusion is unknown. Three papers in this issue of Immunity demonstrate that CD43 exclusion involves the ERM family of cytoskeletal proteins. PMID- 11728331 TI - Resistance is futile: assimilation of cellular machinery by HIV-1. AB - HIV-1 budding appears to require Vps4 and Tsg101-two proteins that have links to endosomal sorting machinery. A picture emerges wherein divergent viruses recruit endosomal proteins like Tsg101 to gain access to ubiquitin processes that play a crucial role during viral budding. PMID- 11728332 TI - Exclusion of CD43 from the immunological synapse is mediated by phosphorylation regulated relocation of the cytoskeletal adaptor moesin. AB - Formation of the immunological synapse requires TCR signal-dependent protein redistribution. However, the specific molecular mechanisms controlling protein relocation are not well defined. Moesin is a widely expressed phospho-protein that links many transmembrane molecules to the cortical actin cytoskeleton. Here, we demonstrate that TCR-induced exclusion of the large sialoprotein CD43 from the synapse is an active event mediated by its reversible binding to moesin. Our results also reveal that relocalization of moesin is associated with changes in the phosphorylation status of this cytoskeletal adaptor protein. Finally, these findings raise the possibility that the change in moesin localization resulting from TCR engagement modifies the overall topology of the lymphocyte membrane and facilitates molecular interactions at the site of presenting cell contact. PMID- 11728333 TI - The GTPase Rac-1 controls cell fate in the thymus by diverting thymocytes from positive to negative selection. AB - The positive selection of CD4 or CD8 single-positive mature peripheral T lymphocytes and the deletion of self-reactive cells are crucial for central tolerance in the peripheral immune system. Previously, the guanine nucleotide binding protein Rac-1 has been shown to control pre-T cell development. The present report now describes the actions of Rac-1 in thymocyte selection. The study reveals that this molecule has the striking and unique ability to efficiently divert cells from positive selection into a pathway of negative selection and deletion. The ability of Rac-1 to switch thymocytes from a destiny of positive to negative selection identifies this molecule as a critical regulator of the developmental processes in T cells that are essential for immune homeostasis. PMID- 11728334 TI - The membrane-microfilament linker ezrin is involved in the formation of the immunological synapse and in T cell activation. AB - Dynamic interactions between membrane and cytoskeleton components are crucial for T cell antigen recognition and subsequent cellular activation. We report here that the membrane-microfilament linker ezrin plays an important role in these processes. First, ezrin relocalizes to the contact area between T cells and stimulatory antigen-presenting cells (APCs), accumulating in F-actin-rich membrane protrusions at the periphery of the immunological synapse. Second, T cell receptor (TCR)-mediated intracellular signals are sufficient to induce ezrin relocalization, indicating that this protein is an effector of TCR signaling. Third, overexpression of the membrane binding domain of ezrin perturbs T cell receptor clustering in the T cell-APC contact area and inhibits the activation of nuclear factor for activated T cells (NF-AT). PMID- 11728335 TI - Distinct patterns of membrane microdomain partitioning in Th1 and th2 cells. AB - Here we show that activated Th1 and Th2 cells have distinct patterns of membrane compartmentalization into lipid rafts. TCR complex members are recruited efficiently to rafts and aggregate with rafts at the site of MHC/peptide contact in Th1 cells but not Th2 cells. TCR/raft association in Th1 cells is deficient in the absence of CD4, suggesting that CD4 aids recruitment of the TCR to rafts. We show differential utilization of rafts in Th1 and Th2 cells by cholesterol depletion studies, which alters calcium signaling in Th1 but not Th2 cells. Furthermore, Th2 cells have a decreased ability to respond to low-affinity peptide stimulation. These studies indicate that components of membrane microdomains are differentially regulated in functionally distinct CD4 T cells. PMID- 11728336 TI - ERM-dependent movement of CD43 defines a novel protein complex distal to the immunological synapse. AB - The large mucin CD43 is actively excluded from T cell/APC interaction sites, concentrating in a membrane domain distal to the site of TCR engagement. The cytoplasmic region of CD43 was necessary and sufficient for this antipodal movement. ERM cytoskeletal adaptor proteins colocalized with CD43 in this domain. An ERM dominant-negative mutant blocked the distal accumulation of CD43 and another known ERM binding protein, Rho-GDI. Inhibition of ERM function decreased the production of IL-2 and IFNgamma, without affecting PKC(theta) focusing or CD69 upregulation. These results indicate that ERM proteins organize a complex distal to the T cell/APC interaction site and provide evidence that full T cell activation may involve removal of inhibitory proteins from the immunological synapse. PMID- 11728337 TI - The immunological synapse of CTL contains a secretory domain and membrane bridges. AB - Cytotoxic T lymphocytes (CTL) rapidly destroy their targets. Here we show that although target cell death occurs within 5 min of CTL-target cell contact, an immunological synapse similar to that seen in CD4 cells rapidly forms in CTL, with a ring of adhesion proteins surrounding an inner signaling molecule domain. Lytic granule secretion occurs in a separate domain within the adhesion ring, maintaining signaling protein organization during exocytosis. Live and fixed cell studies show target cell plasma membrane markers are transferred to the CTL as the cells separate. Electron microscopy reveals continuities forming membrane bridges between the CTL and target cell membranes, suggesting a possible mechanism for this transfer. PMID- 11728338 TI - A critical role for Dnmt1 and DNA methylation in T cell development, function, and survival. AB - The role of DNA methylation and of the maintenance DNA methyltransferase Dnmt1 in the epigenetic regulation of developmental stage- and cell lineage-specific gene expression in vivo is uncertain. This is addressed here through the generation of mice in which Dnmt1 was inactivated by Cre/loxP-mediated deletion at sequential stages of T cell development. Deletion of Dnmt1 in early double-negative thymocytes led to impaired survival of TCRalphabeta(+) cells and the generation of atypical CD8(+)TCRgammadelta(+) cells. Deletion of Dnmt1 in double-positive thymocytes impaired activation-induced proliferation but differentially enhanced cytokine mRNA expression by naive peripheral T cells. We conclude that Dnmt1 and DNA methylation are required for the proper expression of certain genes that define fate and determine function in T cells. PMID- 11728339 TI - Cr2, a candidate gene in the murine Sle1c lupus susceptibility locus, encodes a dysfunctional protein. AB - The major murine systemic lupus erythematosus (SLE) susceptibility locus, Sle1, corresponds to three loci independently affecting loss of tolerance to chromatin in the NZM2410 mouse. The congenic interval corresponding to Sle1c contains Cr2, which encodes complement receptors 1 and 2 (CR1/CR2, CD35/CD21). NZM2410/NZW Cr2 exhibits a single nucleotide polymorphism that introduces a novel glycosylation site, resulting in higher molecular weight proteins. This polymorphism, located in the C3d binding domain, reduces ligand binding and receptor-mediated cell signaling. Molecular modeling based on the recently solved CR2 structure in complex with C3d reveals that this glycosylation interferes with receptor dimerization. These data demonstrate a functionally significant phenotype for the NZM2410 Cr2 allele and strongly support its role as a lupus susceptibility gene. PMID- 11728340 TI - Inhibition of IFN-gamma signaling by an Epstein-Barr virus immediate-early protein. AB - Viruses have evolved elaborate mechanisms to target many aspects of the host's immune response. The cytokine IFN-gamma plays a central role in resistance of the host to infection via direct antiviral effects as well as modulation of the immune response. In this study, we demonstrate that the Epstein-Barr virus (EBV) immediate-early protein, BZLF1, inhibits the IFN-gamma signaling pathway. BZLF1 decreases the ability of IFN-gamma to activate a variety of important downstream target genes, such as IRF-1, p48, and CIITA, and prevents IFN-gamma-induced class II MHC surface expression. Additionally, BZLF1 inhibits IFN-gamma-induced STAT1 tyrosine phosphorylation and nuclear translocation. Finally, we demonstrate that BZLF1 decreases expression of the IFN-gamma receptor, suggesting a mechanism by which EBV may escape antiviral immune responses during primary infection. PMID- 11728341 TI - Constitutive CD27/CD70 interaction induces expansion of effector-type T cells and results in IFNgamma-mediated B cell depletion. AB - The interaction between the TNF receptor family member CD27 and its ligand CD70 provides a costimulatory signal for T cell expansion. Normally, tightly regulated expression of CD70 ensures the transient availability of this costimulatory signal. Mice expressing constitutive CD70 on B cells had higher peripheral T cell numbers that showed increased differentiation toward effector-type T cells. B cell numbers in CD70 transgenic (TG) mice progressively decreased in primary and secondary lymphoid organs. This B cell depletion was caused by CD27-induced production of IFNgamma in T cells. We conclude that apart from its role in controlling the size of the activated T cell pool, CD27 ligation contributes to immunity by facilitating effector T cell differentiation. PMID- 11728342 TI - The IL-7 receptor controls the accessibility of the TCRgamma locus by Stat5 and histone acetylation. AB - The IL-7 receptor (IL-7R) plays critical roles in expansion and V(D)J recombination during lymphocyte development. Here we demonstrate that cytokine stimulation rapidly recruits Stat5 and transcriptional coactivators to the Jgamma germline promoter and induces histone acetylation, germline transcription, and accessibility in Ba/F3 cells. We also show that histone acetylation of the TCRgamma locus is significantly reduced in IL-7R-deficient thymocytes and that the introduction of active Stat5 restores the histone acetylation and accessibility of the locus. Furthermore, treatment with histone deacetylase inhibitor recovers the histone acetylation and accessibility in IL-7R-deficient thymocytes. Therefore, these results suggest that Stat5 may recruit the transcriptional coactivators to the Jgamma germline promoter and control the accessibility of the TCRgamma locus by histone acetylation. PMID- 11728343 TI - Rapid secretion of interleukin-1beta by microvesicle shedding. AB - The proinflammatory cytokine interleukin-1beta (IL-1beta) is a secreted protein that lacks a signal peptide and does not follow currently known pathways of secretion. Its efficient release from activated immune cells requires a secondary stimulus such as extracellular ATP acting on P2X(7) receptors. We show that human THP-1 monocytes shed microvesicles from their plasma membrane within 2-5 s of activation of P2X(7) receptors. Two minutes after such stimulation, the released microvesicles contained bioactive IL-1beta, which only later appeared in the vesicle-free supernatant. We conclude that microvesicle shedding is a major secretory pathway for rapid IL-1beta release from activated monocytes and may represent a more general mechanism for secretion of similar leaderless secretory proteins. PMID- 11728344 TI - A novel TNF receptor family member binds TWEAK and is implicated in angiogenesis. AB - TWEAK is a member of the TNF ligand family that induces angiogenesis in vivo. We report cloning of a receptor for TWEAK (TweakR) from a human umbilical vein endothelial cell (HUVEC) library. The mature form of TweakR has only one hundred and two amino acids and six cysteine residues in its extracellular region. Five different assays demonstrate TWEAK-TweakR binding, and the interaction affinity constant (Kd) is within a physiologically relevant range of 2.3 +/- 0.1 nM. The TweakR cytoplasmic domain binds TRAFs 1, 2, and 3. Cross-linking of TweakR induces HUVEC growth, and mRNA levels are upregulated in vitro by a variety of agents and in vivo following arterial injury. Soluble TweakR inhibits endothelial cell migration in vitro and corneal angiogenesis in vivo. PMID- 11728345 TI - Role of calcium influx in cytotoxic T lymphocyte lytic granule exocytosis during target cell killing. AB - One mechanism cytotoxic T lymphocytes use to kill targets is exocytosis of cytotoxic agents from lytic granules, a process that requires Ca(2+) influx. We investigated the role of Ca(2+) influx in granule exocytosis using TALL-104 human leukemic cytotoxic T cells triggered via a bispecific antibody containing an anti CD3 F(ab') to kill Raji B lymphoma cells. Using a novel fluorescence method, we detected target-directed release of approximately 15% of lytic granules during killing. Consistent with previous work, we observed sustained CTL Ca(2+) gradients during killing, but gradients reflect the behavior of Fura-2 in granules. Rapid imaging experiments suggest that Ca(2+) channels are not polarized during killing, indicating that Ca(2+) influx does not direct granule reorientation. Furthermore, we find that Ca(2+) acts via a high-affinity interaction to promote granule exocytosis. PMID- 11728346 TI - Relation of the leukocyte count to recurrent cardiac events in stable patients after acute myocardial infarction. AB - Increasing evidence implicates inflammation as a risk factor for coronary artery disease. We determined whether an elevated leukocyte count is associated with an increased risk of death or reinfarction in stable patients with a past acute myocardial infarction (AMI). The current analysis is a substudy of the Multicenter Diltiazem Postinfarction Trial, which investigated the effect of diltiazem on mortality and reinfarction in 2,466 patients hospitalized for AMI. We included 1,294 patients in whom a leukocyte count was obtained 6 months after the index AMI. The composite end point of reinfarction or death was used as the primary end point of the study and reinfarction or cardiac death was used as a secondary end point. The study population was divided into 4 quartiles (Q1, Q2, Q3, and Q4) based on the leukocyte count. During a mean follow-up period of 25 months, 163 patients reached the primary end point: 8.7%, 10.9%, 14.0%, and 16.7%, in Q1, Q2, Q3, and, Q4 respectively (p = 0.01). After adjusting for potential covariates, Cox proportional-hazards analysis revealed that an increased leukocyte count was associated with an increased risk of both the primary end point (hazard ratio/1 quartile increase in leukocyte count, 1.26; p = 0.003; 95% confidence interval 1.08 to 1.47) and secondary end point (hazard ratio, 1.18/1-quartile increase; p = 0.05; 95% confidence interval 1.00 to 1.40). In conclusion, an increased leukocyte count measured in the stable post-AMI period is associated with an increased risk of cardiac events. These findings indicate that the leukocyte count may be another marker of an atherosclerotic inflammatory process that contributes to cardiac events in postinfarction patients. PMID- 11728347 TI - Clinical significance of abnormal T waves in patients with non-ST-segment elevation acute coronary syndromes. AB - T-wave abnormalities are common electrocardiographic occurrences in patients with non-ST-segment elevation acute coronary syndromes. Although these abnormalities are considered relatively benign, physicians use them to guide therapies. The study objective was to examine the prognostic predictive information of T-wave abnormalities in the setting of unstable coronary artery disease. The T-wave abnormality criterion was based on a new set of normal T-wave amplitude limits differentiated by gender, age, electrocardiographic lead, and QRS axis. Four hundred sixty-eight patients suspected of an acute ischemic incident and considered ineligible for reperfusion therapy were included. Thirteen categories of T-wave abnormalities were tested prospectively. The primary 30-day end point was the combination of refractory angina, myocardial infarction, or death. Quantitative T-wave analysis in an electrocardiographic core laboratory revealed 6 of 13 prespecified categories of T-wave abnormalities that were significantly associated with an adverse outcome. T-wave abnormalities had no prognostic value when ST-segment depression was also present, but this occurred in only 7.9% of patients. T-wave abnormalities as the sole manifestation of ischemia were common (74.4%). Patients with abnormal T waves in > or =1 of 6 selected abnormality categories (70.3%) had a significantly higher risk of death, acute myocardial infarction, and refractory angina (11% vs 3%; p = 0.018). Thus, T-wave abnormalities in patients presenting with non-ST-segment elevation acute coronary syndromes are common and should not automatically be regarded as benign phenomena. Quantitative T- wave analysis provides optimal risk stratification. PMID- 11728348 TI - Prospective comparison of hemorrhagic complications after treatment with enoxaparin versus unfractionated heparin for unstable angina pectoris or non-ST segment elevation acute myocardial infarction. AB - Patients with unstable angina pectoris (UAP) or non-ST-segment elevation acute myocardial infarction (AMI) are at risk of death or recurrent ischemic events, despite receiving aspirin and unfractionated heparin (UFH). This study investigates the effect of the low molecular weight heparin, enoxaparin, on the incidence of hemorrhage and thrombocytopenia in relation to baseline characteristics and subsequent invasive procedures. Rates of hemorrhage and thrombocytopenia were analyzed for UAP or non-ST-segment elevation AMI in patients included in the prospective, randomized, double-blind Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-wave Coronary Events (ESSENCE) study. Patients received either enoxaparin or UFH, plus aspirin, for 2 to 8 days. The overall rate of major hemorrhage (at 30 days) was comparable between the 2 groups (6.5% for enoxaparin vs. 7.0% for UFH, p = 0.6). The rate of major hemorrhage while on treatment was slightly higher in the enoxaparin group, but this was not significant (1.1% vs 0.7% for UFH, p = 0.204), as was the rate of major hemorrhage within 48 hours of coronary artery bypass grafting performed within 12 hours of treatment. However, the rate of minor hemorrhage was significantly higher in the enoxaparin group, with the majority being injection-site ecchymoses or hematomas (11.9% vs. 7.2% with UFH, p <0.001). Thrombocytopenia (platelet count <100,000 per mm(3)) occurred mainly in association with coronary bypass surgery, with a similar rate in both groups. Thus, enoxaparin is a well-tolerated alternative to UFH in the management of UAP or non-ST-segment elevation AMI. Despite the more effective antithrombotic effect, which results in fewer ischemic events, enoxaparin is not associated with an increase in the rate of major hemorrhagic complications, and is not significantly associated with thrombocytopenia, but is associated with an increase in minor injection site ecchymosis. PMID- 11728349 TI - Usefulness of beat-to-beat QT dispersion fluctuation for identifying patients with coronary heart disease at risk for ventricular arrhythmias. AB - QT dispersion (QTd) has been proposed as a marker of ventricular repolarization inhomogeneity and arrhythmic risk, but despite the dynamic properties of ventricular repolarization and the fact that QTd represents only a static parameter of the repolarization event, beat-to-beat QTd behavior has not been assessed so far. Beat-to-beat QTd was automatically measured in 12-lead digitized electrocardiographic recordings obtained over 10 minutes from 77 subjects. Forty seven patients had coronary heart disease without a history of ventricular arrhythmias, 12 had coronary heart disease and documented ventricular tachycardia, and 18 were normal subjects. Mean QTd, the difference between the maximum minus the minimum QTd observed over the recording time (QTd variation), the maximum difference of QTd between consecutive beats (QTd maximum), and QTd SD (QTd variability) were analyzed between groups. The temporal QTd parameters were significantly increased in the group of patients with susceptibility to ventricular arrhythmias compared with those in both healthy subjects (QTd variation p <0.0001, QTd maximum p <0.001, QTd variability p <0.0001) and patients without a history of ventricular tachycardia (QTd variation p <0.01, QTd maximum p <0.01, QTd variability p <0.01). Mean QTd was also significantly increased in the groups of patients with and without ventricular tachycardia compared with controls (p <0.001 and <0.01), but did not differ significantly between the 2 patient groups. Thus, QTd has a dynamic behavior with significant beat-to-beat fluctuations even in normal subjects and is more marked in patients with coronary heart disease at risk for ventricular arrhythmias. PMID- 11728350 TI - Safety of the weight-adjusted dosing regimen of tenecteplase in the ASSENT-Trial. AB - The results of the ASsessment of Safety and Efficacy of a New Thrombolytic agent (ASSENT-2) trial revealed that tenecteplase (TNK) is equivalent to tissue plasminogen activator (t-PA) for treating myocardial infarction. Because careful consideration of safety is important with all agents, including the newer bolus therapies, and across a range of doses, this study evaluated the safety of TNK compared with t-PA across a range of weight and dose categories. The 5 doses of TNK ranged from 30 to 50 mg and were adjusted for estimated weight. Rates of death and intracranial hemorrhage were determined among patients receiving TNK and t-PA in ASSENT-2, stratified by categories of estimated weight corresponding to each TNK dose. Respective rates of death with TNK versus t-PA were not significantly different in any estimated weight category: <60 kg (12.54% vs 11.46%), 60 to 69 kg (8.22% vs. 8.97%), 70 to 79 kg (5.57% vs 5.48%), 80 to 89 kg (4.66% vs 5.36%), and > or =90 kg (4.91% vs. 3.96%, all p > or =0.26). Respective rates of intracranial hemorrhage were also not significantly different: <60 kg (2.20% vs. 2.29%), 60 to 69 kg (0.97% vs. 1.33%), 70 to 79 kg (1.15% vs. 1.10%), 80 to 89 kg (0.73% vs 0.49%), and > or =90 kg (0.47% vs 0.47%, all p > or =0.33). Adjustment for small baseline differences in this randomized sample did not change the results. Thus, across the range of estimated weight categories corresponding to each TNK dose, TNK is as safe and effective as t-PA. PMID- 11728351 TI - Immediate and long-term clinical and angiographic results from Wiktor stent treatment for true bifurcation narrowings. AB - From January 1996 to December 1998, 90 consecutive patients with true bifurcation lesions underwent percutaneous coronary angioplasty with Wiktor stent implantation in our centers. In 1 group (group I, n = 45), a simple approach (main vessel stenting and balloon angioplasty of the side branch) was pursued. In the other group (group II, n = 45), both the main vessel and the side branch were stented ("T" technique). There was no significant difference in clinical and angiographic characteristics between the 2 groups. Angiographic and procedural successes were 100% and 95.6%, respectively, in both groups. Angiographic results for the side branch were better in group II than in group I. In-hospital and long term (12 month) major cardiac events were similar in the 2 groups. Target lesion revascularization was 15.5% in group I and 35.5% in group II (p = 0.12). In the main vessel, restenosis rate was 12.5% in group I and 25% in group II (p = 0.15). In the side branch, restenosis rate was 37.5% in group II and 12.5% in group I (p = <0.05; odds ratio 2.42; 95% confidence interval 1.05 to 6.26). Event-free probability at 12 months was 61% in group II and 80% in group I (p = 0.10). When dealing with true bifurcation lesions, a simple strategy is associated with a lower risk of restenosis in the side branch. In contrast, a complex approach does not appear to give any benefit in terms of early or long-term outcome or restenosis rate. PMID- 11728352 TI - Development and validation of a clinical prediction rule for major adverse outcomes in coronary bypass grafting. AB - In this study, we develop and internally validate a clinical prediction rule for in-hospital major adverse outcomes, defined as death, renal failure, reinfarction, cardiac arrest, cerebrovascular accident, or coma, in patients who underwent coronary artery bypass grafting (CABG). All adult patients (n = 9,498) who underwent a CABG and no other concomitant surgery at 12 academic medical centers from August 1993 to October 1995 were included in the study. We assessed in-hospital major adverse outcomes and their predictors using information on admission, coronary angiography, and postoperative hospital course. Predictor variables were limited to information available before the procedure, and outcome variables were represented only by events that occurred postoperatively. We developed and internally validated a clinical prediction rule for any major adverse outcome after CABG. The rule's ability to discriminate outcomes and its calibration were assessed using receiver-operating characteristic analysis and the Hosmer-Lemeshow goodness-of-fit statistic, respectively. A major adverse outcome occurred in 6.5% of patients in the derivation set and 7.2% in the validation set. Death occurred in 2.5% of patients in the derivation set and 2.2% in the validation set. Sixteen variables were independently correlated with major adverse outcomes, with the risk score value attributed to each risk factor ranging from 2 to 12 points. The rule stratified patients into 6 levels of risk based on the total risk score. The spread in probability between the lowest and highest risk groups of having a major adverse outcome was 1.7% to 32.3% in the derivation set and 2.2% to 22.3% in the validation set. The prediction model performed well in both outcome discrimination and calibration. Thus, this clinical prediction rule allows accurate stratification of potential CABG candidates before surgery according to the risk of experiencing a major adverse outcome postoperatively. PMID- 11728353 TI - Effect of center volume on outcome of ventricular septal defect closure and arterial switch operation. AB - This study evaluates the effects of surgical volume and preoperative patient characteristics on length of stay following ventricular septal defect (VSD) repair and arterial switch operation (ASO). Twenty-four centers contributed data on 16,795 consecutive surgical procedures from January 1, 1992 to December 31, 1996 to the Pediatric Cardiac Care Consortium Registry. The following variables were used in the analysis: center, center volume, age, weight, date of operation, preoperative hospital days, presence of trisomy 21, and significant coexisting heart defects (risk). For VSD, all variables were associated with length of stay in the univariate analyses; however, in the multivariate model, center volume did not retain significance. Length of stay for ASO was associated with center, center volume, and preoperative days in the univariate analysis. In the multivariate analysis for ASO, center and center volume were significant variables for predicting length of stay. These findings suggest that length of stay for uncomplicated surgical procedures is not related to surgical volume; however, for technically demanding procedures, length of stay tends to be shorter in centers with more experience. PMID- 11728355 TI - James Stuart Forrester III, MD: a conversation with the editor [interview by William Clifford Roberts]. PMID- 11728354 TI - Usefulness of visceral obesity (waist/hip ratio) in predicting vascular endothelial function in healthy overweight adults. AB - Vascular endothelial dysfunction (VED) is associated with obesity; however, its etiology remains controversial. By determining the predictors of fasting and postprandial endothelial function in overweight adults without other cardiovascular risk factors, we were able to investigate novel mechanisms directly linking obesity to VED. Thirty-two healthy adults (body mass index [BMI] > or =27 kg/m(2)) underwent determination of fasting low-density lipoprotein (LDL) particle size, high sensitivity C-reactive protein levels, anthropometric measurements, and endothelial function by flow-mediated dilation (FMD) of the brachial artery. Postprandial lipemia and FMD were measured 4 hours after ingestion of a high-fat meal. Blood pressures and fasting levels of lipoproteins, glucose, insulin, and fatty acids were within normal limits in all subjects. An abdominal fat pattern, as determined by an increased waist/hip ratio (WHR), was the sole significant predictor of FMD (r = -0.58, p = 0.001), despite no significant correlation between whole body obesity (BMI) and FMD. At comparable levels of BMI, obese subjects with a WHR > or =0.85 had a significantly blunted FMD compared with those with a WHR <0.85 (3.93 +/- 2.85% vs 8.34 +/- 5.47%, p = 0.016). Traditional coronary risk factors, C-reactive protein, postprandial lipemia, and LDL particle size did not predict FMD. We found no appreciable alteration in the postprandial state from fasting FMD (6.31 +/- 4.62% vs 6.25 +/- 5.47%, p = 0.95). The same results were found when women were analyzed alone. Increased abdominal adiposity determined by a simple WHR is a strong independent predictor of VED even in healthy overweight adults; this is a finding unexplained by alterations in conventional risk factors, systemic inflammation, or the atherogenic lipoprotein pattern. PMID- 11728356 TI - A risk stratification scheme for selection of a glycoprotein IIb/IIIa inhibitor during percutaneous coronary intervention based on clinical and angiographic criteria. PMID- 11728357 TI - Effect of pravastatin on endothelial function in patients with coronary artery disease (cholesterol-independent effect of pravastatin). PMID- 11728358 TI - Usefulness of worsening clinical status or exercise performance in predicting future events in patients with coronary artery disease. PMID- 11728359 TI - Effect of beta blockers alone, of angiotensin-converting enzyme inhibitors alone, and of beta blockers plus angiotensin-converting enzyme inhibitors on new coronary events and on congestive heart failure in older persons with healed myocardial infarcts and asymptomatic left ventricular systolic dysfunction. PMID- 11728360 TI - Association between obesity and coronary atherosclerosis and vascular remodeling. PMID- 11728361 TI - An improved dobutamine protocol for myocardial perfusion imaging. PMID- 11728363 TI - Comparison of the effects of active left and right pectoral pulse generators on defibrillation efficacy. PMID- 11728362 TI - Rapid effects on vascular function after initiation and withdrawal of atorvastatin in healthy, normocholesterolemic men. PMID- 11728364 TI - Altered atrial, atrioventricular, and ventricular conduction in patients with the long QT syndrome caused by the DeltaKPQ SCN5A sodium channel gene mutation. PMID- 11728365 TI - Effects of losartan in patients with a systemically functioning morphologic right ventricle after atrial repair of transposition of the great arteries. PMID- 11728366 TI - Predicting the identity of decompressing veins after cavopulmonary anastomoses. PMID- 11728367 TI - Effectiveness of shielding for patients during cardiac catheterization or electrophysiologic testing. PMID- 11728368 TI - Pericardial access using the PerDUCER and flexible percutaneous pericardioscopy. PMID- 11728369 TI - Vasovagal susceptibility to nitrate or isoproterenol head-up tilt. PMID- 11728370 TI - Effect of normative aging on midwall left ventricular systolic performance. PMID- 11728371 TI - Effect of spaceflight on cardiovascular responses to upright posture in a 77-year old astronaut. PMID- 11728372 TI - High salt intake, its origins, its economic impact, and its effect on blood pressure. PMID- 11728373 TI - Tissue Doppler-derived myocardial diastolic function distinguishes pathologic and physiologic left ventricular hypertrophy. PMID- 11728374 TI - Detecting aspirin resistance with the platelet function analyzer (PFA-100). PMID- 11728375 TI - Quinapril ischemic event trial. PMID- 11728378 TI - FESTIVAL study is not a festival. PMID- 11728379 TI - Dual inhibitors of cyclooxygenase and 5-lipoxygenase. A new avenue in anti inflammatory therapy? AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are a mainstay in the treatment of inflammatory disease and are among the most widely used drugs worldwide. They are anti-inflammatory, antipyretic, and analgesic and are prescribed as first choice for the treatment of rheumatic disorders and, in general, inflammation. The main limitation in using NSAIDs consists in their side-effects, including gastrointestinal ulcerogenic activity and bronchospasm. The mechanism of action of these drugs is attributed to the inhibition of cyclooxygenase (COX), and, consequently, the conversion of arachidonic acid into prostaglandins. It is hypothesized that the undesirable side-effects of NSAIDs are due to the inhibition of COX-1 (constitutive isoform), whereas the beneficial effects are related to the inhibition of COX-2 (inducible isoform). Arachidonic acid can also be converted to leukotrienes (LTs) by the action of 5-lipoxygenase (5-LOX). LTC(4,) LTD(4,) and LTE(4) are potent bronchoconstrictors, whereas LTB(4) is chemotactic for leukocytes and plays an important role in the development of gastrointestinal ulcers by contributing to the inflammatory process. Thus, developing dual inhibitor compounds that will simultaneously inhibit COX and 5 LOX could enhance their individual anti-inflammatory effects and reduce the undesirable side-effects associated with NSAIDs, especially of the gastrointestinal tract. The most promising COX/5-LOX inhibitor is ML3000 ([2,2 dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3-dihydro-1H-pyrrolizine-5-yl]-acetic acid), now in Phase III clinical trials. This new approach will certainly help to unravel the mechanisms at the root of the undesirable effects of NSAIDs and to develop safer NSAIDs. PMID- 11728380 TI - Induction of G(2)/M phase arrest and apoptosis by a new synthetic anti-cancer agent, DW2282, in promyelocytic leukemia (HL-60) cells. AB - We studied the effect of DW2282-,[(S)-(+)-4-phenyl-1-[N-(4-aminobenzoyl)-indoline 5-sulfonyl-4,5-dihydro-2-imidazolone].hydrochloride], a newly developed anti cancer agent, on cell proliferation, cell cycle progression, and induction of apoptosis in human promyelocytic leukemia (HL-60) cells. DW2282, a diarylsulfonylurea compound, was cytotoxic to HL-60 cells, with an IC(50) of 1.0 microg/mL. Treatment with DW2282 fragmented DNA in a concentration- and time dependent manner, suggesting that these cells underwent apoptosis. Flow cytometric analysis further confirmed that DW2282-treated HL-60 cells were hypodiploid, in terms of DNA content, and were arrested at the G(2)/M phase. The cell cycle arrest was reversible upon the removal of DW2282. HL-60 cells also underwent distinct morphological changes in response to DW2282 treatment, including the appearance of elongated cells with conical tails and other apoptotic characteristics. G(2)/M phase cell cycle arrest was accompanied by a decrease in the levels of cdc2, a protein that plays a critical role for progression through the G(2)/M phase. Treatment of HL-60 cells with DW2282 was also associated with decreased levels of the anti-apoptotic protein Bcl-2, activation of caspase-3, and proteolytic cleavage of poly(ADP-ribose) polymerase. Taken together, these results demonstrate that DW2282 dramatically suppressed HL 60 cell growth by inducing apoptosis after G(2)/M phase arrest. These findings are consistent with the possibility that G(2)/M phase arrest was mediated by the down-regulation of cdc2 levels in HL-60 cells. The data also suggest that DW2282 triggered apoptosis by decreasing Bcl-2 levels and activating caspase-3 protease. These results provide important new information towards understanding the mechanisms by which DW2282 and other diarylsulfonylureas mediate their therapeutic effects. PMID- 11728381 TI - Suppression of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated CYP1A1 and CYP1B1 induction by 12-O-tetradecanoylphorbol-13-acetate: role of transforming growth factor beta and mitogen-activated protein kinases. AB - The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) enhances or suppresses the transcriptional activation of CYP1A1 by 2,3,7,8-tetrachlorodibenzo p-dioxin (TCDD) in a cell/tissue-specific manner. The basis for these effects is not known. Exposure of the immortalized human breast epithelial cell line MCF10A Neo to TPA at the time of, or up to 12 hr prior to, the addition of TCDD strongly suppressed the transcriptional activation of CYP1A1 and CYP1B1 (IC(50) approximately 0.5 nM). A recent study (Carcinogenesis 2000;21:1303-12) demonstrated that TPA-treated MCF10A-Neo cells rapidly activate the latent transforming growth factor beta (TGFbeta) in the serum used to supplement the culture medium. The suppressive effects of TPA on CYP1A1 induction by TCDD in MCF10A-Neo cultures could be partially suppressed by: (a) co-incubation of TCDD + TPA-treated cultures with a neutralizing TGFbeta pan antibody; (b) prior removal of latent TGFbeta from the culture medium; or (c) switching cultures to serum- and growth factor-free medium immediately before the addition of TPA and TCDD. Exposure of cultures to TPA 24-48 hr prior to subsequent TPA + TCDD treatment not only inhibited the suppressive effects of TPA, but markedly enhanced CYP1A1 mRNA accumulation. TPA caused a rapid and protracted activation of extracellular signal-regulated kinases (ERKs). Pretreatment of cultures with the mitogen activated protein kinase kinase (MEK) inhibitor PD184352 [2-(2-chloro-4-iodo phenylamino)-N-cyclopropyl-methoxy-3,4-difluoro-benzamide] completely inhibited ERK activation by TPA. However, PD184352 did not prevent the suppressive effects of TPA on CYP1A1 activation by TCDD. These studies demonstrate that TPA initiates protein kinase C-dependent, ERK-independent processes that suppress CYP1A1 activation by TCDD in MCF10A-Neo cells. Furthermore, TGFbeta mediates a small portion of this suppressive activity. PMID- 11728382 TI - Positive modulation by Ras of interleukin-1beta-mediated nitric oxide generation in insulin-secreting clonal beta (HIT-T15) cells. AB - In the present study, we have shown that exposure of insulin-secreting clonal beta (HIT-T15) cells to interleukin-1beta (IL-1beta) results in a time- and concentration-dependent increase in nitric oxide (NO) release. These effects by IL-1beta on NO release were mediated by induction of inducible nitric oxide synthase (iNOS) from the cells. Preincubation of HIT cells with Clostridium sordellii lethal toxin-82, which irreversibly glucosylates and inactivates small G-proteins, such as Ras, Rap, Ral, and Rac, but not Cdc42, completely abolished IL-1beta-induced NO release. Pre-exposure of HIT cells to C. sordellii lethal toxin-9048, which monoglucosylates and inhibits Ras, Cdc42, Rac, and Rap, but not Ral, also attenuated IL-1beta-mediated NO release. These data indicate that activation of Ras and/or Rac may be necessary for IL-1beta-mediated NO release. Preincubation of HIT cells with C. difficile toxin-B, which monoglucosylates Rac, Cdc42, and Rho, had no demonstrable effects on IL-mediated NO release, ruling out the possibility that Rac may be involved in this signaling step. Further, two structurally dissimilar inhibitors of Ras function, namely manumycin A and damnacanthal, inhibited, in a concentration-dependent manner, the IL-1beta mediated NO release from these cells. Together, our data provide evidence, for the first time, that Ras activation is an obligatory step in IL-1beta-mediated NO release and, presumably, the subsequent dysfunction of the pancreatic beta cell. Our data also provide a basis for future investigations to understand the mechanism of cytokine-induced beta cell death leading to the onset of insulin dependent diabetes mellitus. PMID- 11728383 TI - Accompanying protein alterations in malignant cells with a microtubule polymerizing drug-resistance phenotype and a primary resistance mechanism. AB - Microtubules (MTs) are cytoskeletal components whose structural integrity is mandatory for the execution of many basic cell functions. Utilizing parental and drug-resistant ovarian carcinoma cell lines that have acquired point mutations in beta-tubulin and p53, we studied the level of expression and modification of proteins involved in apoptosis and MT integrity. Extending previous results, we demonstrated phosphorylation of pro-survival Bcl-x(L) in an epothilone-A resistant cell line, correlating it with drug sensitivity to tubulin-active compounds. Furthermore, Mcl-1 protein turned over more rapidly following exposure to tubulin-modifying agents, the stability of Mcl-1 protein paralleling the drug sensitivity profile of the paclitaxel or epothilone-A resistant cell lines. The observed decreases in Mcl-1 were not a consequence of G(2)M arrest, as determined by flow cytometry analysis, which showed prominent levels of Mcl-1 in the absence of any drug treatment in populations enriched in mitotic cells. We also observed that a paclitaxel-resistant cell line expressed Bax at a much lower level than the sensitive parental line [A2780(1A9)], consistent with its mutant p53 status. MT-associated protein-4 (MAP4), whose phosphorylation during specific phases of the cell cycle reduces its MT-polymerizing and -stabilizing capabilities, was phosphorylated in response to drug challenge without a change in expression. Phosphorylation of MAP4 correlated with sensitivity to tubulin-binding drugs and with a dissociation from MTs. We propose that the tubulin mutations, which result in a compromised paclitaxel:tubulin or epothilone:tubulin interaction and paclitaxel or epothilone resistance, indirectly inhibit downstream events that lead to cell death, and this, in turn, may contribute to the drug-resistance phenotype PMID- 11728384 TI - Reduction in serotonin synthesis following acute and chronic treatments with paroxetine, a selective serotonin reuptake inhibitor, in rat brain: an autoradiographic study with alpha-[14C]methyl-L-tryptophan(2). AB - Serotonin (5-HT) synthesis rates were calculated on the basis of the assumption that trapping of alpha-[14C]methyl-L-tryptophan (alpha-[14C]MTrp) is directly related to brain 5-HT synthesis. In the first series of experiments, an acute intraperitoneal injection of paroxetine (10 mg/kg) produced a significant reduction in 5-HT synthesis in brain structures containing serotonergic cell bodies (the dorsal, median, and pallidum raphe nuclei), as well as in most projection areas: the ventral tegmental area, median forebrain bundle, hippocampus CA3 region, and nigrostriatal structures (substantia nigra, lateral and medial caudate nuclei). The reductions in the projection areas were greater (between 25 and 53%) than in those areas containing serotonergic cell bodies (between 18 and 23%). In the cerebral cortex, 5-HT synthesis rates were not modified by acute paroxetine treatment. In a second series of experiments, rats were treated with paroxetine (10 mg/kg/day, s.c., delivered by osmotic minipumps) for 14 days. There was a marked decrease (39-69%) in 5-HT synthesis in every structure examined. In conclusion, the present data suggest that the effects of paroxetine on 5-HT synthesis in the cerebral cortex are different from its effects in the cell body area of the brainstem. PMID- 11728385 TI - Enhancing effect of zinc on astroglial and cerebral endothelial histamine uptake. AB - We have studied the effect of zinc ion on the uptake of histamine (HA) into cultured astroglial and cerebral endothelial cells and established that Zn(2+) enhances the uptake of the amine dose-dependently and in remarkable extents by increasing the V(max) to about 3-fold (from 3.25 +/- 0.42 to 8.50 +/- 0.97 pmol/mg protein/min in astroglial cells) without altering the K(M) (0.20 +/- 0.03 microM) significantly. The stimulatory effect of zinc ion showed strong sensitivity for VUF 8407, an inhibitory compound of astroglial and cerebral endothelial uptake of HA. In the presence of 20 microM VUF 8407 the zinc-enhanced uptake was reduced by about 50% in both cell types. Binding measurements revealed increased capacities of the zinc-exposed HA binding (B(max)= 0.41 +/- 0.05 increased to 1.21 +/- 0.16 pmol/mg protein in astroglial membranes and B(max) = 0.25 +/- 0.03 enhanced to 1.05 +/- 0.12 pmol/mg protein in cerebral endothelial membranes) but statistically unchanged affinity of the ligand for HA carrier (K(D) values calculated as 35.2 +/- 3.4 nM and 45.1 +/- 3.8 nM for astroglial bindings; whereas 25 +/- 2.1 nM and 30 +/- 2.6 nM for cerebral endothelial bindings of the amine). The compound VUF 8407 reduced the B(max) of zinc-exposed HA binding of astroglial membranes but did not modify the K(D) of the zinc exposed membrane significantly. The ex vivo experiments confirmed our in vitro findings; an i.c.v. dose of 0.4 micromol/kg ZnSO(4,) 24 hr after the injection, enhanced the uptake of [(3)H]HA into dissociated hypothalamic and cerebellar cells to about 2- and 3-fold, respectively. Present data clearly showed that zinc exposures enhance the astroglial and the cerebral endothelial uptake of HA in vitro and it might be considered that zinc produces similar effects in vivo. Free zinc may participate in the regulation of the extraneuronal HA concentration and this metal ion (endogenous or exogenous) might be favored in the removal of the amine from the interstitial space especially in conditions with relatively high HA. PMID- 11728386 TI - Retinoid X receptor agonist elevation of serum triglycerides in rats by potentiation of retinoic acid receptor agonist induction or by action as single agents. AB - Hypertriglyceridemia is a major side-effect of retinoid therapy in humans. We previously reported that agonists for the retinoic acid receptors (RARs), but not the retinoid X receptors (RXRs), elevate serum triglycerides in male Fischer rats, and that, surprisingly, the RAR/RXR pan-agonists 9-cis-retinoic acid and AGN 191659 [(E)-5-[2-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen 1-yl]-2-thiophenecarboxylic acid] induce 2- to 3-fold higher levels of serum triglycerides than the RAR-selective agonists alone. We have now demonstrated that hypertriglyceridemia induced by an RAR agonist, AGN 190121 [4-[4-(2',6',6' trimethylcyclohex-1-enyl)-but-1-yne-3-enyl]benzoic acid], is substantially potentiated by the RXR-selective agonists AGN 191701 [(E) 2-[2-(5,6,7,8 tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen-1-yl]-4-thiophene-carboxylic acid] and AGN 192849 [(3,5,5,8,8,-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl) (5 carboxypyrid-2-yl)sulfide] in a dose-dependent manner. RXR-specific retinoids, as previously reported, had no independent effect on serum triglycerides when tested at 24 hr after final dosing, but did elicit a reversible hypertriglyceridemia at 2.5 and 5 hr. This induction of serum triglycerides could not be blocked by the potent RAR-specific antagonist AGN 193109 [4-[(5,6-dihydro 5,5-dimethyl-8-(4-methylphenyl)-2-naphthalenyl)-ethynyl] benzoic acid]. The RXR ligand-induced hypertriglyceridemia was independent of the effect of feeding or fasting. The relative potencies of RXR-specific retinoids for acute triglyceride elevation (AGN 194204 [3,7-dimethyl-6S,7S-methano-7-[1,1,4,4-tetramethyl-1,2,3,4 tetrahydronaphth-7-yl] 2(E),4(E) heptadienoic acid] > AGN 192849 approximately AGN 191701) approximately correlated with potencies in the activation of the RXR receptors. The RAR/RXR pan-agonist effect included >50% inhibition of total heparin-releasable lipase activity in serum, consistent with inhibition of lipase mediated triglyceride disposal. These data also indicate that RAR and RXR ligands can act synergistically to induce hypertriglyceridemia through distinct mechanisms of action. PMID- 11728387 TI - Expression of rat aldehyde reductase AKR7A1: influence of age and sex and tissue specific inducibility. AB - The regulation of the aldo-keto reductase AKR7A1 was examined in the livers of male and female rats during development by using Western blots, and its contribution to carbonyl metabolism was assessed by using enzyme assays. Hepatic levels of AKR7A1 are low in fetal rats and rise to a peak at around 6 weeks of age in animals of both sexes. Higher levels of the enzyme are found in adult male rat liver than in adult female rat liver. The reductase, therefore, appears to be subject to sex-specific regulation. The effect of growth hormone in mediating this difference in expression was examined by using hypophysectomized animals whose serum growth hormone levels had been feminized by continuous administration. Results demonstrate that such treatment leads to a reduction in AKR7A1 expression. AKR7A1 was found to be constitutively expressed in rat tissues such as liver, kidney, small intestine, and testis, but it was not detected in nasal mucosa, skeletal muscle, heart, adrenal gland, brain, or spleen. However, AKR7A1 was inducible by the synthetic antioxidant ethoxyquin in liver, kidney, and small intestine, but not in the other tissues examined. These results show that levels of this important detoxication enzyme vary considerably according to age and sex and that dietary antioxidants can also influence its level in several tissues. PMID- 11728388 TI - Inhibitory properties of the anti-bothropic complex from Didelphis albiventris serum on toxic and pharmacological actions of metalloproteases and myotoxins from Bothrops asper venom. AB - Anti-bothropic complex (ABC) was isolated from the serum of the South American opossum (Didelphis albiventris) by single-step affinity chromatography using a Sepharose-immobilized metalloprotease (BaP1) from Bothrops asper as the binding protein. Biochemical characterization of ABC showed the presence of two glycosylated subunits of 43 and 45 kDa, respectively, with an isoelectric point < 4. The two subunits were separated by ion-exchange HPLC. The N-terminal sequences of both subunits (LKAMDPTPXLWIETESP, where X is Arg-9 and Pro-9, respectively) showed a high degree of identity with other serum inhibitors isolated from different marsupials. Functional studies pointed out that ABC inhibits the hemorrhagic and proteolytic activities on fibrin, fibrinogen, and casein induced by the metalloproteases BaP1 and BaH4 isolated from B. asper venom. In addition to the anti-hemorrhagic and anti-proteolytic activities, ABC also showed anti myotoxic, anti-lethal, and anti-edematogenic effects against myotoxic phospholipases A(2) isolated from the same venom. Moreover, it had inhibitory effects on the phospholipase A(2) activity of the crude venom as well as the isolated venom phospholipases A(2). PMID- 11728389 TI - Synthesis and pharmacological activity of a novel water-soluble hepatocyte specific polymeric prodrug of prostaglandin E(1) using lactosylated poly(L glutamic hydrazide) as a carrier. AB - A novel polymeric prodrug of prostaglandin E(1) (PGE(1)) was synthesized using lactosylated poly(L-glutamic hydrazide) (Lac-NH-PLGA) as a targetable carrier to hepatocytes. Poly(L-glutamic hydrazide) (PLGA-HZ) was prepared by reacting poly(gamma-benzyl-L-glutamate) with hydrazine monohydrate, followed by coupling with lactose via a hydrazone linkage. Then the lactosylated PLGA-HZ was reduced by sodium cyanoborohydride (NaBH(3)CN) in order to make the linkage irreversible (Lac-NH-PLGA). Finally, PGE(1) was bound to hydrazide moieties remaining in Lac NH-PLGA without any condensing agent under weakly acidic conditions (pH 5) where PGE(1) would be chemically most stable at room temperature (PGE(1) conjugate). The PGE(1) conjugate prepared was sufficiently water-soluble in spite of the hydrophobic nature of its backbone (-NH-CH-CO-) and PGE(1) itself. After intravenous injection in mice, the [111In]PGE(1) conjugate rapidly accumulated in the liver, whereas [111In]PLGA-HZ did not, suggesting the involvement of a galactose-specific mechanism in the uptake of the [111In]PGE(1) conjugate. Fractionation of liver cells revealed that the [111In]PGE(1) conjugate was preferentially taken up by liver parenchymal cells. The pharmacological activity was examined in mice with fulminant hepatitis induced by intraperitoneal injection of carbon tetrachloride. Intravenous injection of the PGE(1) conjugate at a dose of 1 mg (0.065 mg PGE(1))/kg effectively inhibited the increase in plasma glutamic pyruvic transaminase (GPT) activity compared with that of free PGE(1) at a dose of 0.065 or 0.65 mg/kg. These results suggest that the PGE(1) conjugate is an excellent prodrug for the treatment of fulminant hepatitis. PMID- 11728390 TI - Effects of nitric oxide and hydrogen peroxide on histamine release from RBL-2H3 cells. AB - We have studied the effect of nitric oxide (NO) and hydrogen peroxide (H(2)O(2)), two reactive oxygen species (ROS) on histamine release (HR) from RBL-2H3 cells, a rat mucosal-type mast cell line. Marked HR was elicited by antigen (DNP-HSA), calcium ionophore A23187, sodium fluoride or phospholipase C, but not with compound 48/80 or 1,2-dioctanoyl-sn-glycerol. The NO-synthase substrate L arginine and its inactive enantiomer (D-arginine), each on its own, induced a small but significant increase in HR above the basal level. However, the NO donors (sodium nitroprusside or NaNO(3)) or the NO-synthase inducer lipopolysaccharide did not induce HR. Moreover, methylene blue (MB), which inhibits guanylate cyclase and N(omega)-nitro-L-arginine (L-NA), an inhibitor of NO synthase, were also without effect on either the basal HR or the L-arginine induced HR. HR induced by A23187, DNP-HSA, sodium fluoride or phospholipase C was markedly reduced by MB, but mildly by L-NA (both at 1-100 microM). H(2)O(2) (0.01 1.0 mM) on its own did not induce HR, but it had a potent inhibitory effect on DNP-HSA- or A23187-induced HR, which was not reversed by L-NA (1-100 microM). Taken together, it seems that neither the stimulatory nor the inhibitory effects of the NO-related compounds on HR can be attributed to NO, but rather to other mechanisms. The inhibition of HR by H(2)O(2) also does not involve NO and suggests a negative feedback regulatory role for the peroxide in the allergic inflammation. PMID- 11728391 TI - Effect of atorvastatin, simvastatin, and lovastatin on the metabolism of cholesterol and triacylglycerides in HepG2 cells. AB - We evaluated the effects of the hydroxymethylglutaryl coenzyme A reductase inhibitors (HMGRI) atorvastatin, lovastatin, and simvastatin on lipid homeostasis in HepG2 cells. The drugs were almost equally effective in inhibiting cholesterol synthesis and in decreasing cellular cholesterol. Atorvastatin and lovastatin increased low-density lipoprotein receptor mRNA (2.5-fold at 3 x 10(-7) M) and the transcription rate at the promoter of the low-density lipoprotein receptor gene (>5-fold at 10(-6) M). The three compounds enhanced the activity of the low density lipoprotein receptor at a similar magnitude (1.6-2.1- fold at 10(-6) M). Atorvastatin and lovastatin increased the nuclear form of sterol regulatory element binding protein (SREBP)-2, but not of SREBP-1. Each of the drugs increased triacylglyceride synthesis (50% at 10(-7)-10(-6) M), cellular triacylglyceride content (16% at 10(-6) M), and expression of fatty acid synthase by reporter gene and Northern blot analysis (2-fold and 2.7-fold at 10(-6) M and 3 x 10(-7) M, respectively). All compounds reduced the secretion of apo B (30% at 3 x 10(-7) M). HMGRI decreased the ratio of cholesterol to apo B in newly synthesised apo B containing particles by approximately 50% and increased the ratio of triacylglycerides to apo B by approximately 35%. We conclude that regulatory responses to HMGRI are mediated by SREBP-2 rather than by SREBP-1, that HMGRI oppositely affect the cellular cholesterol and triacylglyceride production, that HMGRI moderately decrease the release of apo B containing particles, but profoundly alter their composition, and that atorvastatin does not significantly differ from other HMGRI in these regards. PMID- 11728392 TI - An efficient and stereoselective synthesis of beta-D-Arap-(1-->2)-beta-D-Galp-(1- >3)-beta-D-Galp-(1-->4)-alpha-D-Manp, a tetrasaccharide fragment of Leishmania major lipophosphoglycan. AB - A tetrasaccharide fragment of Leishmania major lipophosphoglycan (which seems to be involved in a biological mechanism for the parasite transmission) has been synthesised using the thioglycoside, trichloroacetimidate and halide-exchange glycosylation procedures and step-wise chain elongation strategy. PMID- 11728393 TI - Synthesis, characterization and the first crystal structure of the Zn(II) complex of 4,6-O-ethylidine-N-(2-hydroxybenzylidene)-beta-D-glucopyranosylamine. AB - 4,6-O-Ethylidine-N-(2-hydroxybenzylidene)-beta-D-glucopyranosylamine (H(3)L(1)) and N-(5-bromo-2-hydroxybenzylidene-4,6-O-ethylidine-beta-D-glucopyranosylamine (H(3)L(2)) molecules possessing a single bond C-1 single bond N double bond C(H) single bond moiety for metal-ion binding were synthesized by condensing the 4,6-O ethylidene-beta-D-glucopyranosylamine with salicylaldehyde or 5 bromosalicylaldehyde. Complexes of these ligands with Zn(II) were isolated and characterized using elemental analysis, FTIR, UV-Vis absorption, NMR spectroscopic and FAB mass spectrometric techniques. The structure of the Zn(II) complex derived from H(3)L(1) was established for the first time by a single crystal X-ray diffraction study. The anomeric nature of the saccharide moiety was established based on (1)H NMR studies and was confirmed by the crystal structure. Further, the structure and binding aspects of the ligand, and the coordination features of this in its Zn(II) complex were derived from the corresponding crystal structure. PMID- 11728394 TI - Synthesis of novel deoxy lambda(5) phospha sugar nucleosides: 1,3-dipolar cycloaddition of an azidophospholane with alkynes. AB - Several novel phospha sugar nucleosides, analogs of normal sugar nucleosides, were synthesized from a phospholene 1-oxide derivative. Bromination of a phospholene precursor in aqueous organic medium gave regio diastereomers, the threo and erythro bromohydrins 3 (1-bromo-1,3,4-trideoxy-1,4-C-[(R,S) phenylphosphinylidene]-glycero-tetrofuranose). Further substitution of the threo isomer 3a with sodium azide led to its corresponding azidophospholane 4 (1-azido 1,3,4-trideoxy-2-methyl-1,4-C-[(R)-phenylphosphinylidene]-beta-D-glycero tetrofuranose). 1,3-Dipolar cycloaddition of 4 with various electron-deficient and electron-rich alkynes afforded triazole derivatives that are nucleoside analogues. The strong electron-withdrawing phosphoryl group in the hemiacetal ring exerted no effect over reaction regioselectivity of the 1,3-dipolar cycloaddition, but steric effects of the alkynes played a vital role on the selectivity, since the regioisomer ratios and the rates and yields of cycloadducts changed as the bulkiness of the substituents on the acetylene changes. Structures of all compounds were unequivocally confirmed by 1H, 13C, and 31P NMR and mass spectral studies. Single crystal X-ray crystallographic analysis of some derivatives allowed determination of configuration of the phospha sugar nucleosides. PMID- 11728395 TI - Syntheses of imido-substituted glycosans and their photocyclisation towards highly functionalised heterotricycles. AB - Reaction of O-protected amino-1,6-anhydro-beta-D-hexopyranoses with succinic or glutaric anhydride and subsequent intramolecular acylation afforded the succinimido- and glutarimido-substituted glycosans. Irradiation with UV light of 254 nm wavelength led to gamma-hydrogen abstraction at the pyranose ring by the excited carbonyl function. The stereoselective recombination of the resulting 1,4 diradicals gave annelated azetidinols, which fragmented by a retrotransannular ring opening reaction to give the glycosan-annelated azepanedione and azocanedione systems, respectively. PMID- 11728396 TI - Generation of anti-factor Xa active, 3-O-sulfated glucosamine-rich sequences by controlled desulfation of oversulfated heparins. AB - In the framework of a project aimed at generating heparin-like sulfation patterns and biological activities in biotechnological glycosaminoglycans, different approaches have been considered for simulating the alpha(1-->4)-linked 2-O sulfated L-iduronic acid (IdoA2SO(3))-->N,6-O-sulfated D-glucosamine (GlcNSO(3)6SO(3)) disaccharide sequences prevalent in mammalian heparins. Since the direct approach of sulfating totally O-desulfated heparins, taken as model compounds for C-5-epimerized sulfaminoheparosan (N-deacetylated, N-sulfated Escherichia coli K5 polysaccharide), preferentially afforded heparins O-sulfated at C-3 instead than at C-2 of the iduronate residues, leading to products with low anticoagulant activities, the problem of re-generating a substantial proportion of the original IdoA2SO(3) residues was circumvented by performing controlled solvolytic desulfation (with 9:1 v/v DMSO-MeOH) of extensively sulfated heparins. The order of desulfation of major residues of heparin GlcN and IdoA and of the minor one D-glucuronic acid was: GlcNSO(3)>GlcN6SO(3)>IdoA3SO(3) congruent with GlcA2SO(3) congruent with GlcN3SO(3)>IdoA2SO(3) congruent with GlcA3SO(3). Starting from a 'supersulfated' low-molecular weight heparin, we obtained products with up to 40% of iduronate residues O-sulfated exclusively at C-2 and up to 40% of their glucosamine residues O-sulfated at both C-6 and C-3. Upon re-N-sulfation, these products displayed an in vitro antithrombotic activity (expressed as anti-factor Xa units) comparable with those of current low molecular weight heparins. PMID- 11728397 TI - Preparation and characterisation of fluorescent chitosans using 9-anthraldehyde as fluorophore. AB - Chitosans with chemical composition ranging from a fraction of N-acetylated units (F(A)) of 0.01 to 0.61 were used to prepare fluorescence labelled chitosans by reductive amination with 9-anthraldehyde. Fluorescent chitosans with a low theoretical degree of substitution (DS, 0.001-1%) were prepared, and the actual DS of the products were determined by UV and (1)H NMR spectroscopy. The fluorescence excitation and emission spectra of the chitosan with F(A) of 0.09 and DS 1% showed an excitation maximum at 254 nm and an emission maximum at 413 nm. The intrinsic viscosities ([eta]) of the fluorescent chitosans were compared to those of the original chitosans, showing that the derivatisation procedure lead only to a negligible decrease in [eta]. The conformation of these fluorescent chitosans with very low DS-values is not altered and they can conveniently be directly quantified by UV or fluorescence spectroscopy. PMID- 11728398 TI - Two stereoisomeric 3(I),2(II)-anhydro-alpha-cyclodextrins: a molecular dynamics and crystallographic study. AB - Regioselective epoxide ring opening of 2(I),3(I)-(2(I)S)-anhydro-alpha cyclodextrin (1) through intramolecular attack of hydroxyl groups of neighboring glucose rings occurs in diequatorial fashion to yield 3(I),2(II)-anhydro-alpha cyclodextrin (3) with a rigid glucopyranose-dioxane-glucopyranose tricyclic ring system, the usual diaxial opening and the gluco/altro-configured stereoisomer 2 cannot be detected. Molecular dynamic simulations in water were used to analyze the conformations of 1-3 and the stereochemical implications of this reaction. Due to the contracted 2,3-OH side of the torus, 3 features an inverted conicity compared to the parent alpha-cyclodextrin. A crystallographic study on the bis 3.3 n-PrOH nonahydrate not only displays little variations between the solid state and solution geometries of 3, but also provides a molecular picture of a unique inclusion complex in which three n-propanol molecules are distributed in the cavity of a dimeric unit of 3 (monoclinic, space group P2(1), a=14.257(1), b=22.623(2), c=16.644(1) A, beta=104.82(1) degrees, all 19,278 reflections with I>2 sigma(I) yield R(F)=0.1017). PMID- 11728399 TI - Preparative route to glucuronyl donors bearing temporary protecting group at O-3 via 6,3-lactonisation by Bz(2)O or Piv(2)O. AB - Heating of non-substituted beta-D-glucopyranuronic acids in the presence of pivalic or benzoic anhydride in DMF gives 4-O-monoacylated or 2,4-di-O-acylated 6,3-lactones that can be easily transformed into corresponding methyl uronates bearing free OH-groups at C-2 and C-3 or C-3 only, respectively. This method was used for preparation of selectively 3-O-protected glucuronyl donors of thioglycoside and trichloracetimidate types. PMID- 11728400 TI - Synthesis of polyhydroxyindolizidines from 5,6-dihydro-2H-pyran-2-one. AB - (1aR,5aR,5bS,6S,7S)-6,7-Di-tert-butoxy-5-oxo-pyrrolidino[1,2-b]isoxazolidino[4,5 c]tetrahydropyran (8) prepared by (1,3)-dipolar cycloaddition of the cyclic nitrone 6 derived from tartaric acid to 5,6-dihydro-2H-pyran-2-one (7) was transformed into indolizidine 11 via a sequence of reactions involving methanolysis of the lactone ring, intramolecular alkylation of the nitrogen atom promoted by a carbontetrabromide-triphenylphosphine mixture and hydrogenolysis of the N single bond O bond. Decarboxylation of 11 provided known 7 hydroxylentiginosine derivative 14, whereas oxidative decarboxylation gave indolizidine 15 structurally related to castanospermine. PMID- 11728401 TI - (1)H NMR studies of hydroxy protons of the V[beta-Gal(1-->3)-alpha-GalNAc(1- >O)]THPGY glycopeptide. AB - The hydroxy protons of the disaccharide moiety in the glycopeptide Val-[beta Gal(1-->3)-alpha-GalNAc(1-->O)]-Thr-His-Pro-Gly-Tyr (1) have been investigated in aqueous solution using (1)H NMR spectroscopy. The chemical shifts (delta), coupling constants ((3)J(CH,OH)), temperature coefficients (d delta/dT), exchange rates (k(ex)), and NOEs have been measured. The data show that the O(2')H of Gal has a reduced contact with water due to steric interference caused by the 2 acetamido group of GalNAc. No interaction, in terms of hydrogen bonding exists between the disaccharide and the peptide moieties, but the rotation around the sugar-peptide linkage is restricted. PMID- 11728402 TI - Chemical structure of a polysaccharide isolated from the cell wall of Arachniotus verruculosus and A. ruber. AB - The structure of a cell wall alkali-extractable and water-soluble polysaccharide isolated from two species of Arachniotus has been established by reductive cleavage and NMR spectroscopy. The linear polysaccharide consists of a regular disaccharide-repeating unit with the structure: [-->6)-beta-D-Galf-(1-->5)-beta-D Galf-(1-->](n)-->mannan core. PMID- 11728403 TI - Structural heterogeneity in the lipopolysaccharides of Pseudomonas syringae with O-polysaccharide chains having different repeating units. AB - Studies by sugar and methylation analyses, Smith degradation, and 1H and 13C NMR spectroscopy revealed a structural heterogeneity in the O-polysaccharides of Pseudomonas syringae pvs. coronafaciens IMV 9030 and atrofaciens IMV 8281 owing to the presence of different types of repeating units. In strain IMV 9030, the major repeating units are a linear alpha-L-rhamnose trisaccharide and a tetrasaccharide (A, n=0 or 1). A minor repeating unit is a branched pentasaccharide with an alpha-L-rhamnose main chain and a lateral 3-acetamido-3,6 dideoxy-D-galactose (D-Fuc3NAc) residue (B, X=2, n=1). In strain IMV 8281, all repeating units are branched and differ in size and position of substitution of one of the alpha-L-rhamnose residues (tetrasaccharide, B, X=3, n=0; pentasaccharides, B, X=2 or 3, n=1). [structure--see text] Reinvestigation of the structure of the branched O-polysaccharide of P. syringae pv. tomato IPGR 140 showed that, together with the major tetrasaccharide repeating unit (B, X=3, n=0) [Knirel, Y. A., et al. Carbohydr. Res. 1993, 243, 199-204], it has a minor pentasaccharide repeating unit (B, X=3, n=1). PMID- 11728405 TI - Introduction to "Screening for Disease". PMID- 11728406 TI - Screening for the early detection of disease, the need for evidence. AB - BACKGROUND: Screening for the early detection of disease has had a spotted history. Structured approaches to the process of gathering and evaluating evidence, with the emphasis on well-controlled randomized studies, have greatly improved the beneficial potential of appropriate and effective screening. Good quality evidence will contribute to quality health care. ISSUES: The volunteer participants in screening programs must give fully informed consent. This means that they must be presented with clear and accurate statements of the advantages and disadvantages of the screening program. Among the screening programs that have been conducted include hyperhomocysteinemia and coronary artery disease, Down's syndrome, Neonatal Group B streptococcal disease, Type 2 diabetes mellitus and endometrial cancer. The evidence in these studies has strengths and weaknesses as to how they support or oppose a particular intervention. The laboratory has a major role to play in establishing and validating standards of accuracy for diagnostic tests. Agreement on standards and their application does not mean the end of different interpretation and controversy. CONCLUSIONS: Laboratory physicians and scientists will be very effective consultants if they have the best available, high quality evidence for the appropriate use of laboratory tests. PMID- 11728407 TI - Objective evaluation of data in screening for disease. AB - BACKGROUND: In order to decide on the feasibility of screening for disease by laboratory testing, a number of assessments need to be made. The necessary procedures and statistical calculations are required for an objective evaluation of the diagnostic accuracy of a laboratory test and of the potential for screening and of the data obtained from the screening. METHODS: The determination of reference intervals, is an essential first step in the evaluation of the performance of a laboratory test. The procedures and calculations are necessary for the determination of the sensitivity, specificity, diagnostic predictive value and receiver-operating characteristic curve of a test and their respective applications in laboratory screening for disease. The additional statistical tools include odds ratios and likelihood ratios for assessing the performance of a laboratory test. CONCLUSION: These tools are used in the development of an appropriate clinical decision limit in a defined population of subjects and under defined conditions for acceptable screening. PMID- 11728408 TI - Economic evaluations of screening programs: a review of methods and results. AB - BACKGROUND: The economics of screening is part of the broader area of the economics of prevention, and is concerned with the allocation of resources to screening activities when confronted with the problem of scarcity. ISSUES: One important aspect of the economics of screening is the economic evaluation of screening programs. Economic evaluation involves a systematic assessment of the costs and benefits of screening. RESULTS: A review of methods and results of economic evaluations of screening produces no generalizable conclusion about the economic desirability or otherwise, of screening programs. Each particular screening program must be evaluated on its merits. There is no evidence of any systematic tendency for screening programs to be more or less cost-effective than other prevention or treatment programs. Across all programs, substantial disparities have been found in cost-effectiveness. CONCLUSIONS: The challenge in the future will be to secure the re-allocations of resources necessary to maximize health gains from limited health care resources, rather than to undertake the analyses on which such re-allocations would be based. PMID- 11728409 TI - Some ethical and design challenges of screening programs and screening tests. AB - BACKGROUND: There are many examples of inadequate screening studies because of design faults and consent problems. ISSUES: Informed consent is the right of each person engaged in a clinical interaction, including service and research involvement in presymptomatic screening. Any screening approach must answer questions of efficacy, effectiveness, availability and efficiency. Accurate information, presented in clear language, must state a realistic assessment of the limitations, harms and benefits of the screening, including the consequences of false positive and false negative tests. There must be no coercive element in obtaining consent. Written and oral information should be presented. It is also important to recognize cultural differences between societies that emphasize individual autonomy and those in which autonomy derives from membership of a family, group or community, thus requiring different strategies to protect the vulnerable. Testing for genetic disorders presents additional challenges, partly due to the presence of relatively small numbers of affected individuals in the population, but also because of the complexity of counseling, confidentiality issues and the potential for social, insurance or employment discrimination. CONCLUSIONS: It is becoming increasingly difficult to present the data for many screening procedures in ways that can be understood by possible participants, and at the same time deal clearly with the costs to society and the efficacy and efficiency of the tests. PMID- 11728410 TI - Screening for cardiovascular disease with cholesterol. AB - BACKGROUND: Total cholesterol (TC) and lipoprotein measurements are among the major risk factors for cardiovascular disease (CVD). ISSUES: The massive (and increasing) disease burden associated with cardiovascular disorders justifies public health measures, such as screening, which may assist with the identification and treatment of individuals who have a high risk of developing these disorders. Whole populations are placed at risk by environmental factors such as dietary habits and exercise patterns, and this may be reflected by simple tests such as TC that largely depend on dietary consumption of saturated fat and cholesterol. On the other hand, identification of high-risk individuals within populations requires more specific tests such as measurement of cholesterol in lipoprotein fractions. This recognizes that the contribution of different risk factors may vary between individuals. Dietary and pharmacological interventions, particularly those that reduce low-density lipoprotein cholesterol (LDL-C), have been shown to prevent the onset or recurrence of CVD. CONCLUSIONS: It is desirable and justifiable to try to identify high-risk patients before the onset of clinical CVD because morbidity and mortality may occur as a consequence of the initial event. The relationship between the costs and the benefits of prophylactic interventions varies according to the extent to which clinical procedures, including laboratory testing, can estimate the level of cardiovascular risk. It can be argued that inappropriate cholesterol screening may have a negative effect on health economics and patient attitudes, but these problems appear easily surmountable. Techniques that enable the preclinical detection of CVD may help the process to strike a balance between public health initiatives that reduce the environmental factors responsible for the epidemic of CVD, and the strategies that are known to reduce event rates in high-risk individuals. PMID- 11728411 TI - Screening for diabetes: is it warranted? AB - BACKGROUND: Diabetes mellitus is estimated to have a worldwide prevalence of 4.6% and afflict 200 million people. The prevalence is accelerating rapidly and the disease has reached epidemic proportions. While type 1 diabetes usually has a dramatic clinical onset, almost half of all those individuals with type 2 diabetes have not been diagnosed. This observation, coupled with the presence of complications at diagnosis and the significant reduction of microvascular complications obtained with tight glycemic control, has led to the recommendation that screening for diabetes be instituted. The American Diabetes Association recommends that adults aged 45 years or more should be evaluated for diabetes by measuring fasting plasma glucose concentrations. CONCLUSIONS: The rationale for screening, evidence that tight glycemic control reduces complications, and the possible roles of Hb A(1c) and autoantibodies in screening strategies are addressed in this review. PMID- 11728412 TI - Screening for prostate cancer with prostate-specific antigen: beware the biases. AB - BACKGROUND: The evidence relating to the use of prostate-specific antigen (PSA) as a screening test is a highly controversial, as demonstrated by the lack of agreement among experts. There may be biases associated with various studies. ISSUES: The main controversy is the relatively high prevalence of prostate cancer (PC) found at autopsy compared with the relatively low death rate from the disease. The lack of modifiable risk factors has led to early detection as a strategy to reduce mortality, as there is evidence for a significant burden of disease. Important issues are the accuracy of current screening tests, some attempts to improve on them, and whether there are good prognostic markers. The consequences of PSA testing (usually further testing including biopsy) and outcomes of treatment are presented in terms of mortality and morbidity; quality of life (QOL) must also be considered. Also important are the benefits from, and the difficulties associated with the "informed choice" approach to PSA screening. CONCLUSION: There is evidence to suggest that biases can have a significant impact on the utility of PSA as a screening test for PC. PMID- 11728413 TI - Neonatal biochemical screening for disease. AB - BACKGROUND: The practice of screening the neonatal population for certain diseases by biochemical testing of a dried blood spot is an established public health initiative in many countries. The diseases for which screening is done vary from region to region, based on ethnic, financial and political considerations. Criteria have been established to identify diseases suitable for neonatal screening. DISEASES SCREENED: In Western countries, screening for phenylketonuria (PKU) and congenital hypothyroidism (CH) has been introduced throughout. Subsequently, cost-benefit analysis has confirmed the existence of a financial benefit. Other diseases screened for in some regions include galactosemia, aminoacidemias and organic acidemias, cystic fibrosis, congenital adrenal hyperplasia, biotinidase deficiency, hemoglobinopathies, glucose-6 phosphate dehydrogenase deficiency, and Duchenne muscular dystrophy, although in no case has a clear financial benefit been established. CONCLUSIONS: Since the introduction of neonatal screening over 40 years ago, new methods have resulted in an increase in the number of diseases amenable to screening, better automation and greater specificity. Methods currently used include growth of an inhibited bacterial auxotroph (the original phenylalanine (phe) method of Dr. Robert Guthrie), spectrophotometry, fluorometry, immunoassay, and tandem mass spectrometry with electrospray ionization. PMID- 11728414 TI - Case finding and screening strategies for thyroid dysfunction. AB - BACKGROUND: A case can be made for routine testing for thyroid dysfunction (TD) in women aged over 50 years once every 5 years when they present for medical care (case finding). This recommendation is based on: (i) the prevalence of TD, predominantly hypothyroidism, (ii) the insensitivity of standard clinical assessment in detecting even overt TD, (iii) the sensitivity of a single test, serum thyroid stimulating hormone (TSH), in identifying both over- and under function, (iv) the probable adverse consequences of failure to recognize even mild TD, (v) the safety and effectiveness of treatment, and (vi) presumed lack of adverse effect from the testing program. METHODS: A normal serum TSH value has a high negative predictive value in ruling out primary TD; if TSH is abnormal, measurement of serum free thyroxine (T(4)) and further clinical assessment are both required to establish the degree of TD. RESULTS: Case finding identifies more "subclinical" or mild TD (abnormal TSH, normal T(4) and triiodothyronine (T(3))), than overt disease, but a major benefit of widespread testing is the earlier detection and treatment of unsuspected overt disease. There is now evidence that mild TD has adverse consequences and should not merely be regarded as a prognostic indicator, but there is still no consensus whether there is a causal relationship between mild thyroid failure and dyslipidemia. CONCLUSIONS: A case can be made for treatment of both mild thyrotoxicosis and hypothyroidism, but the therapeutic decision is generally simpler for hypothyroidism. PMID- 11728415 TI - Urine drug screening in the medical setting. AB - BACKGROUND: The term drug screen is a misnomer since it implies screening for all drugs, which is not possible. Current practice is to limit the testing to the examination of serum for several drugs such as ethanol, acetaminophen, salicylate, and of urine for several specific drugs or classes of drugs. In the emergency setting the screen should be performed in less than one hour. Controversies continue to exist regarding the value of urine drug testing in the medical setting. The reasons for these include the drugs involved, the sample, the methods utilized to perform the tests, and the level of understanding of the physician using the data, all of which are closely related to the other. METHODS: Current automated methods provide rapid results demanded in emergency situations, but are often designed for, or adapted from, workplace testing and are not necessarily optimized for clinical applications. Furthermore, the use of these methods without consideration of the frequency in which the drugs are found in a given area is not cost-effective. The laboratory must understand the limitations of the assays used and provide this information to the physician. Additionally, the laboratory and the physicians using the data must cooperate to determine which drugs are appropriate and necessary to measure for their institution and clinical setting. In doing so it should be remembered that for many drugs, the sample, urine, contains the end product(s) of drug metabolism, not the parent drug. Furthermore, it is necessary to understand the pharmacokinetic parameters of the drug of interest when interpreting data. Finally, while testing for some drugs may not appear cost-effective, the prevention or reduction of morbidity and mortality may offset any laboratory costs. CONCLUSIONS: While the literature is replete with studies concerning new methods and a few regarding physician understanding, there are none that we could find that thoroughly, objectively, and fully addressed the issues of utility and cost-effectiveness. PMID- 11728416 TI - Pharmacogenetic screening and therapeutic drugs. AB - BACKGROUND: Pharmacogenetics is the science of the influence of heredity on pharmacological response. ISSUES: The cost of severe adverse drug reactions in individuals has been estimated in the US alone to be in excess of US$4 billion. It has been argued that in a significant proportion of cases, the efficacy and toxicity profiles of drug therapy would be substantially improved in individuals if characteristics due to genetic variation were taken into account. Methods are now available, which make screening for susceptibility feasible. CONCLUSIONS: There are several therapeutic areas in which screening may give rise to significant improvements in outcome with cost-benefits to both the individual and the community. However, there is currently a lack of data on which cost-benefit analysis can be based. The challenge is to provide this information for new drugs, and for drugs with established therapeutic roles. PMID- 11728417 TI - Stool screening for colorectal cancer: evolution from occult blood to molecular markers. AB - BACKGROUND: Colorectal cancer is the second leading cause of malignant death, and better preventive strategies are needed. Participation rates for colorectal cancer screening remain low due, in part, to perceived discomfort, potential harm, and high costs with available tools. METHODS: Stool testing, unlike other conventional screening approaches, is noninvasive and requires no cathartic preparation. However, widely used fecal blood tests yield frequent false-negative and false-positive results that lower screening effectiveness and raise program costs. There is a compelling biological rationale to target DNA alterations exfoliated from neoplasms into stool, and multiple DNA markers would need to be assayed because of the genetic heterogeneity of colorectal neoplasia. Early clinical studies with this multi-target DNA-based stool assay approach suggest high sensitivity for both colorectal cancer and premalignant adenomatous polyps while maintaining high specificity. CONCLUSIONS: This apparently accurate and user-friendly new approach holds promise to improve the effectiveness, efficiency, and appeal of colorectal cancer screening. Large-scale clinical studies are clearly warranted to corroborate the early results. PMID- 11728418 TI - Screening for hemochromatosis. AB - BACKGROUND: Hereditary hemochromatosis is the most common autosomal recessive disorder in populations of northern European descent. ISSUES: Many experts consider hemochromatosis to be an almost ideal disease for population screening because it essentially fulfills almost all the criteria for screening proposed by the WHO. However, others disagree and suggest that more data are required particularly with regard to the natural history and penetrance of the disease. There is also disagreement about the best diagnostic/screening test for the disease and the performance of these tests in the context of screening. Other concerns are the variability and lack of standardization in screening test measurements, the selection of screening threshold values and the identification of false positive cases. The advent of a genetic test for the condition has brought other worries with regard to informed consent and the ethical, legal and social implications of screening particularly in relation to medical and general discrimination. Other important issues include compliance, cost effectiveness and the evidence that screening has lessened the burden of disease in the community. CONCLUSIONS: At the present time, we believe that further data regarding both the exact disease burden and the outcomes of screening studies particularly in the general community are required before widespread population screening is introduced. PMID- 11728419 TI - Desensitization of endogenously expressed delta-opioid receptors: no evidence for involvement of G protein-coupled receptor kinase 2. AB - The involvement of G protein-coupled receptor kinase 2 (GRK2) in the agonist induced desensitization of delta-opioid receptor-mediated inhibition of cAMP formation in NG108-15 mouse neuroblastomaxrat glioma hybrid cells was investigated. Pretreatment of wild-type cells with the delta-opioid receptor agonist [D-Pen(2,5)]-enkephalin (DPDPE; 100 nM) for as little as 5 min produced marked desensitization of subsequent DPDPE-mediated inhibition of iloprost (300 nM)-stimulated cAMP formation. In NG108-15 cells stably overexpressing wild-type GRK2 or dominant negative mutant GRK2 (DNM GRK2), the DPDPE-induced desensitization of cAMP inhibition was the same as in plasmid-transfected control cells. Pretreatment of wild-type cells with the inhibitors of receptor internalization, concanavalin A (0.25 mg ml(-1)) or hypertonic sucrose (0.4 M), also failed to inhibit DPDPE-mediated desensitization. Finally, in NG108-15 cells stably overexpressing G protein-coupled receptor kinase 6 (GRK6), DPDPE-induced desensitization was significantly increased as compared to plasmid-transfected control cells. These results indicate that GRK2 is unlikely to mediate the desensitization of endogenous delta-opioid receptors in NG108-15 cells, but that other GRKs, such as GRK6, may be more important. PMID- 11728420 TI - Role of protein kinase C in opioid modulation of glycine-gated Cl(-) current in rat periaqueductal gray neuron. AB - The Role of protein kinase C in the modulatory effect of a mu-opioid receptor agonist, [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]enkephalin (DAMGO), on the glycine-gated Cl(-) current was examined in acutely dissociated rat periaqueductal gray neurons. Using the nystatin-perforated patch-clamp technique, the neurons were voltage-clamped at -60 mV. The glycine-gated Cl(-) current (I(Gly)) was sensitive to strychnine. On pretreatment with 1 microM DAMGO, the 30-microM glycine response increased with time and showed a maximum amplitude of 209+/-37% of control. After a protein kinase C activator, phorbol-12-myristate-13-acetate (PMA, 0.1 microM) as pretreatment, I(Gly) increased to 138+/-6% of control. The DAMGO potentiation of I(Gly) was not altered by coapplication with PMA. Although protein kinase C inhibitors, chelerythrine (3 microM) and 2-[1-(3 dimethylaminopropyl)indol-3-yl]-3-(indol-3-yl) maleimide (GF109203X, 1 microM), did not alter I(Gly), the DAMGO-induced potentiation of I(Gly) was reduced to 161+/-21% or 164+/-31% of the control after coapplication with chelerythrine or GF109203X, respectively. These results indicate that the potentiation of I(Gly) by a mu-opioid receptor agonist is partly mediated by activation of protein kinase C. PMID- 11728421 TI - Glycyrrhizin and some analogues induce growth of primary cultured adult rat hepatocytes via epidermal growth factor receptors. AB - We investigated the effects of glycyrrhizin (GL-1) and some analogues on DNA synthesis and proliferation in serum-free primary cultures of adult rat hepatocytes. The hepatocytes underwent DNA synthesis and proliferation in response to GL-1 and some analogues. The effects of these agents occurred in a time- and dose-dependent manner. The proliferative potency as judged by half maximal effective concentrations was in the following order: 18-beta-H glycyrrhetinic acid (GL-3; 4.5 x 10(-9) M)<18-beta-H-glycyrrhizin (GL-1; 4.4 x 10(-8) M)<18-alpha-H-glycyrrhetinic acid (GL-6; 6.0 x 10(-8) M). The analogue 18 alpha-H-glycyrrhetinic acid 3-O-beta-D-monoglucuronide (GL-5; 1.0 x 10(-7) M) weakly stimulated hepatocyte DNA synthesis and proliferation, whereas 18-alpha-H glycyrrhizin (GL-4) and 18-beta-H-glycyrrhetinic acid 3-O-beta-D-monoglucuronide (GL-2) did not. The growth-promoting effects of GL-1, GL-3 and GL-6 were significantly inhibited at higher initial plating densities (7.0 x 10(4) and 10 x 10(4) cells/cm(2)). A monoclonal antibody against epidermal growth factor (EGF) receptor (1-100 ng/ml), but not that against EGF (1-100 ng/ml), dose-dependently inhibited glycyrrhizin- and analogue-induced hepatocyte DNA synthesis and proliferation. Specific inhibitors of growth-related signal transducers, such as genistein, PD98059 (2'-amino-3'-methoxyflavone) and rapamycin, completely blocked glycyrrhizin- and analogue-induced hepatocyte DNA synthesis and proliferation. Treatment of hepatocytes with GL-1, GL-3 and GL-6 rapidly stimulated tyrosine phosphorylation of the EGF receptor and p42 MAP kinase, which were inhibited by genistein and PD98059, respectively. These results suggest that glycyrrhizin and some analogues are primary hepatocyte mitogens that bind to EGF receptors and subsequently stimulate the receptor tyrosine kinase/mitogen-activated protein kinase pathway to induce hepatocyte DNA synthesis and proliferation. PMID- 11728422 TI - Endothelin-1 stimulates cardiomyocyte injury during mitochondrial dysfunction in culture. AB - To understand the pathophysiological role of endothelin-1 in the failing heart, we constructed a cellular mitochondrial impairment model and demonstrated the effect of endothelin-1. Primary cultured cardiomyocytes from neonatal rats were pretreated with rotenone, a mitochondrial complex I inhibitor, and the cytotoxic effect of endothelin-1 on the cardiomyocytes was demonstrated. Rotenone gradually decreased the pH of the culture medium with incubation time and caused slight cell injury. Endothelin-1 markedly enhanced the effect of rotenone that decreased the pH of the medium and enhanced cellular injury. The enhancement of the decrease in pH and cell injury induced by endothelin-1 was counteracted by the endothelin ET(A) receptor antagonist BQ123 or by maintaining the pH of the medium by the addition of 50 mM HEPES. Endothelin-1 markedly increased the uptake of 2 deoxyglucose and lactic acid production when the cardiomyocytes were pretreated with rotenone. These findings suggest that the stimulation of glucose uptake and anaerobic glycolysis followed by the increase in lactic acid accumulation in cardiomyocytes under the condition of mitochondrial impairment may be involved, at least in part, in the cellular injury by endothelin-1. Moreover, these findings suggest the possibility that the effect of endothelin-1 on myocardium is reversed by the condition of the mitochondria, and endogenous endothelin-1 may deteriorate cardiac failure with mitochondrial dysfunction. This may contribute to clarify the beneficial effect of endothelin receptor blockade in improving heart failures. PMID- 11728423 TI - Solubilisation and immunoprecipitation of rat striatal adenosine A(2A) receptors. AB - In the present study, we have sought to solubilise adenosine A(2A) receptors from rat striatal membranes using a variety of different detergents. Of the detergents tested, 1% CHAPS (3-[(3-deoxycholic acid (cholamidopropyl) dimethylammonio]-1 propanesulfonate) yielded optimal conditions for solubilisation (in the presence of 3 mg/ml protein, 44% of receptor was solubilised, 50% of total protein was solubilised). An antipeptide antibody was raised against a 15 amino-acid sequence within the predicted third intracellular loop region of the human and rat adenosine A(2A) receptor. The antibody was coupled to protein A immobilised on sepharose CL-4B and used to immunoprecipitate adenosine A(2A) receptors from solubilised rat striatal preparations. Radioligand-binding studies were performed using the selective adenosine A(2) antagonist [(3)H]ZM 241385 (4-(2-[7-amino-2-(2 fury1)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol). Using [(3)H]ZM 241385, the pharmacology of immunoprecipitated adenosine A(2A) receptors was composed to striatal membrane bound adenosine A(2A) receptors and detergent solubilised adenosine A(2A) receptors. [(3)H]ZM 241385 labelled a single saturable binding site with high affinity in all three preparations (membrane bound K(d) 2.7 nM+/-1.0; solubilised K(d) 1.9 nM+/-0.3; immunoprecipitated K(d) 2.2 nM+/-0.7). Additionally, all three assays confirmed a rank order of potency for displacers consistent with adenosine A(2A) receptor pharmacology: ZM 241385>KW 6002 ((E)-8-[2-(3,4-dimethoxyphenyl)ethynyl]-1-3-diethyl-3,7-dihydro-7 methyl-1-purine 2,6 dione)>CGS 21680, (2-(4-(2 carboxyethyl)phenylethylamino)-5' N-ethylcarboxamidoadenosine)>DPCPX (8-cyclopentyl-1,3-dipropylxanthine). We conclude that we have solubilised and immunoprecipitated adenosine A(2A) receptors from rat striatum and that their pharmacology is consistent with native striatal adenosine A(2A) receptors. PMID- 11728424 TI - Influence of neurosteroids on the development of rapid tolerance to ethanol in mice. AB - Our recent study demonstrated that neurosteroids might either facilitate or block chronic tolerance to the incoordinating effects of ethanol. The present study investigated the effects of neurosteroids on the development of rapid tolerance to ethanol-induced motor impairment using the N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine [(+)-MK-801] or the gamma-aminobutyric acid (GABA) type A (GABA(A)) receptor agonist muscimol. Male Swiss mice were pretreated with pregnenolone sulfate (0.03 to 0.15 mg/kg) or dehydroepiandrosterone sulfate (0.05 to 0.20 mg/kg) before administration of ethanol (1.9 or 2.25 g/kg) and tested with the rota-rod apparatus. Twenty-four hours later, all animals were re-tested with the rota-rod after receiving the same dose of ethanol. Pretreatment with pregnenolone sulfate or with dehydroepiandrosterone sulfate significantly facilitated the acquisition of tolerance. However, the administration of (+)-MK-801 reversed the stimulatory action of pregnenolone sulfate but did not affect the actions of dehydroepiandrosterone sulfate on ethanol tolerance. Pretreatment with pregnenolone sulfate or dehydroepiandrosterone sulfate prevented the inhibitory action of muscimol on tolerance development. Taken together, our results suggest that neurosteroids may stimulate the development of rapid tolerance to ethanol and that GABA(A) and NMDA receptor systems may be involved in these actions. PMID- 11728425 TI - SCH 57790, a selective muscarinic M(2) receptor antagonist, releases acetylcholine and produces cognitive enhancement in laboratory animals. AB - The present studies were designed to assess whether the novel muscarinic M(2) receptor antagonist 4-cyclohexyl-alpha-[4[[4-methoxyphenyl]sulphinyl]-phenyl]-1 piperazineacetonitrile (SCH 57790) could increase acetylcholine release in the central nervous system (CNS) and enhance cognitive performance in rodents and nonhuman primates. In vivo microdialysis studies show that SCH 57790 (0.1-10 mg/kg, p.o.) produced dose-related increases in acetylcholine release from rat hippocampus, cortex, and striatum. SCH 57790 (0.003-1.0 mg/kg) increased retention times in young rat passive avoidance responding when given either before or after training. Also, SCH 57790 reversed scopolamine-induced deficits in mice in a passive avoidance task. In a working memory operant task in squirrel monkeys, administration of SCH 57790 (0.01-0.03 mg/kg) improved performance under a schedule of fixed-ratio discrimination with titrating delay. The effects observed with SCH 57790 in behavioral studies were qualitatively similar to the effects produced by the clinically used cholinesterase inhibitor donepezil, suggesting that blockade of muscarinic M(2) receptors is a viable approach to enhancing cognitive performance. PMID- 11728426 TI - Rapid tolerance to Delta(9)-tetrahydrocannabinol and cross-tolerance between ethanol and Delta(9)-tetrahydrocannabinol in mice. AB - Motor incoordination in the rota-rod test was used to assess the development of rapid tolerance to Delta(9)-tetrahydrocannabinol and rapid cross-tolerance between ethanol and Delta(9)-tetrahydrocannabinol in mice. Further, the influence of the cannabinoid receptor antagonist SR 141716A (N-(piperidin-1-yl)-5-(4 chlorophenyl)-4-methyl-1H-pyrazole-3-carboxyamide) on the motor impairment induced by both drugs was examined. Mice were injected on day 1 with equipotent doses of Delta(9)-tetrahydrocannabinol (28 mg/kg, i.p.) and ethanol (2.25 g/kg, i.p.) and tested at 30, 60 and 90 min after the injections. On day 2, control groups received ethanol or Delta(9)-tetrahydrocannabinol, some groups received the same treatment as the day before, while the remaining groups switched the treatment. All groups were tested to evaluate tolerance. The development of rapid tolerance to Delta(9)-tetrahydrocannabinol was observed and pretreatment with ethanol resulted in rapid cross-tolerance to Delta(9)-tetrahydrocannabinol. SR 141716A (2 mg/kg, i.p.) failed to block the development of rapid tolerance to both drugs, ethanol and Delta(9)-tetrahydrocannabinol. These results suggest that Delta(9)-tetrahydrocannabinol, similarly to ethanol, can induce rapid tolerance to motor incoordination in mice. They also support the use of the 2-day protocol as an effective procedure to reduce the length of drug exposure necessary to induce tolerance. PMID- 11728427 TI - beta-Adrenoceptor blockade enhances the anticonvulsant effect of glutamate receptor antagonists against maximal electroshock. AB - In this study, we evaluated whether beta-adrenoceptor antagonists may modify the protective efficacy of dizocilpine (MK-801), a NMDA receptor antagonist, and 1-(4 aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine (GYKI 52466), a non-NMDA (AMPA/kainate) receptor antagonist, against maximal electroshock-induced seizures in mice. Propranolol, acebutolol, metoprolol and atenolol were used in doses that did not alter the electroconvulsive threshold. Propranolol potentiated the anticonvulsant activity of MK-801 and GYKI 52466, significantly lowering their ED(50) values from 0.38 and 15.0 to 0.15 (P<0.001) and 8.4 mg/kg (P<0.001), respectively. Similarly, metoprolol lowered the ED(50) of MK-801 and GYKI 52466 from 0.38 and 15.0 to 0.17 (P<0.05) and 11.2 mg/kg (P<0.05). Acebutolol enhanced the protective action of GYKI 52466, lowering its ED(50) value from 15.0 to 12.2 mg/kg (P<0.05), but not that of MK-801. Atenolol, not penetrating the blood-brain barrier, did not affect the anticonvulsive efficacy of MK-801 and GYKI 52466. In conclusion, beta-adrenoceptor antagonists may act synergistically with excitatory amino acid receptor antagonists to inhibit generalised tonic-clonic seizures. PMID- 11728428 TI - Dynamics of histamine H(3) receptor antagonists on brain histamine metabolism: do all histamine H(3) receptor antagonists act at a single site? AB - Thioperamide, the prototypical histamine H(3) receptor antagonist, acts at the brain histamine H(3) autoreceptor to promote the release and metabolism of neuronal histamine, resulting in higher brain levels of the metabolite tele methylhistamine. However, unlike thioperamide, several new histamine H(3) receptor antagonists enter the central nervous system (CNS), block brain histamine H(3) receptors and increase histamine release without increasing brain tele-methylhistamine levels. Experiments were performed presently in an attempt to understand these results. Consistent with previous findings, thioperamide significantly increased the content and synthesis rate of tele-methylhistamine in mouse and rat brain. In contrast, the histamine H(3) receptor antagonists GT-2227 (4-(6-cyclohexylhex-cis-3-enyl)imidazole) and clobenpropit did not affect tele methylhistamine synthesis rate in mouse whole brain. The histamine H(3) receptor ligand GT-2016 (5-cyclohexyl-1-(4-imidazol-4-ylpiperidyl)pentan-1-one) had no effect on tele-methylhistamine levels in any rat brain region and decreased tele methylhistamine synthesis rates in the mouse whole brain. To examine the possibility that these histamine H(3) receptor antagonists might prevent the methylation of newly released histamine, they were co-administered with thioperamide to determine their effects on the thioperamide-induced stimulation of tele-methylhistamine synthesis. GT-2016 significantly reduced the thioperamide induced activation of tele-methylhistamine synthesis in mouse whole brain and in several regions of rat brain. Although further clarification is needed, these results suggest that some histamine H(3) receptor antagonists may promote the release of neuronal histamine, but also act to reduce histamine methylation in vivo by an unknown mechanism. PMID- 11728429 TI - Beneficial effect of the sigma(1) receptor agonist PRE-084 against the spatial learning deficits in aged rats. AB - Sigma(1) (sigma(1)) receptor agonists showed anti-amnesic properties in pharmacological amnesia models. We now investigated whether the selective sigma(1) receptor agonist, 2-(4-morpholinoethyl)-1-phenylcyclohexane-1 carboxylate hydrochloride (PRE-084), could ameliorate spatial learning in aged animals, using a water-maze procedure. Wistar rats, 3 or 24 months old, were trained to locate a visible platform and then an invisible platform. Finally, a transfer test was performed during saline or PRE-084 treatment. Aged, but not adult, animals showed learning deficits unrelated to visual impairments. The PRE 084 treatment allowed aged animals to learn the new platform location, in terms of decreased latencies to the platform during training and increased presence in the quadrant during retention. The results of these experiments suggest a potential of selective sigma(1) receptor agonists as cognitive enhancers during ageing. PMID- 11728430 TI - Vasorelaxant response to isoprenaline, nitric oxide donor, calcitonin gene related peptide and vasoactive intestinal peptide in aortic rings of adult C57BL/6J mice. AB - The mouse and tissues from this species are increasingly used as experimental models because of the wide variety of gene deletions and overexpressions available in this species. Yet, very little is known about normal vascular responses in the mouse. We investigated the vasorelaxant responses on thoracic aortic rings from the adult male C57BL/6J mouse. Isoprenaline, acetylcholine, calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP) and sodium nitroprusside all caused concentration-dependent relaxations in aortic rings possessing healthy endothelium and precontracted with phenylephrine. Maximum relaxations were 64.9+/-2.6%, 66.8+/-2.9%, 114.3+/-4.6%, 65.1+/-4.2% and 116.2+/-5.1% with -logEC(50) values of 6.76+/-0.14, 7.04+/-0.11, 8.53+/-0.14, 8.29+/-0.26 and 8.10+/-0.20 for isoprenaline, acetylcholine, CGRP, VIP and sodium nitroprusside, respectively. There were significantly smaller responses to isoprenaline, acetylcholine, CGRP and VIP when the endothelium was denuded. The maximum relaxations for isoprenaline, CGRP and acetylcholine were 48.3+/-5.1%, 99.6+/-4.4% and 5.7+/-1.6% with -logEC(50) values of 6.44+/-0.40 and 8.23+/ 0.192, respectively, following endothelium removal. The response to VIP was completely dependent to endothelium. Without precontraction, isoprenaline, at the higher doses, caused small contractions. These experiments provide new information about vascular responses of five vasodilators in aortic rings of adult male C57BL/6J mice. PMID- 11728431 TI - Evidence for presynaptic cannabinoid CB(1) receptor-mediated inhibition of noradrenaline release in the guinea pig lung. AB - Using neurochemical method, evidence was obtained that cannabinoid CB(1) receptors are localized on noradrenergic terminals and their stimulation by WIN 55,212-2 reduces the release of [3H]noradrenaline evoked by axonal activity in a frequency-dependent manner. At stimulation rates of 1 and 3 Hz, there was significant inhibition of noradrenaline release, with IC(50) of WIN-55,212-2 41.5+/-2.6 and 320.5+/-28.2 nM, for 1 and 3 Hz, respectively. Cannabinoid CB(1) receptor antagonist SR 141716A completely prevented WIN-55,212-2 from reducing the release. The release of noradrenaline is negatively modulated by presynaptic alpha(2)-adrenoceptors. Because BRL-44408, an alpha(2B)-adrenoceptor, and prazosin, an alpha(1)- and alpha(2B)-adrenoceptor antagonist, both increased the release of [(3)H]noradrenaline, it seems likely that the alpha(2B) subtype is responsible for the negative feedback modulation of noradrenaline release. In the presence of alpha(2)-adrenoceptor antagonism, cannabinoid CB(1) receptor activation by WIN-55,212-2 was much more effective in inhibiting the release of [(3)H]noradrenaline. Using a specific antibody against the C-terminus of the rat cannabinoid CB(1) receptor and also against neuropeptide Y, ultrastructural evidence was obtained that cannabinoid CB(1) receptors are exclusively localized on neuropeptide Y-positive noradrenergic varicosities. Since the sympathetic innervation of the human airway smooth muscle is sparse, and mainly the circulating adrenaline relaxes the airways via activation of beta(2)-adrenoceptor localized on the smooth muscle, it is suggested that inhibition of noradrenaline release by cannabinoids, and the subsequent bronchospasm, may be limited to those cases when noradrenaline released from sympathetic varicosities is involved in airway relaxation. PMID- 11728432 TI - Role of prostaglandin E(2) and Ca(2+) in bradykinin induced contractions of guinea-pig gallbladder in vitro. AB - In this study, we investigated the contribution of prostaglandin E(2) to bradykinin induced contractions of guinea-pig gallbladder in vitro and characterized the sources of activator Ca(2+) for the bradykinin mediated contractions. Contractions induced by bradykinin in guinea-pig gallbladder smooth muscle strips were significantly attenuated by the cyclooxygenase inhibitor piroxicam (10 microM). In the presence of piroxicam, a threshold concentration of prostaglandin E(2) (1 nM) significantly enhanced the contractile response to subsequent challenge with bradykinin. Contractile responses to bradykinin were abolished in a Ca(2+)-free medium plus EDTA. The inhibitor of receptor mediated Ca(2+) entry, SK&F 96365 (1-[beta-[3-(4-methoxyphenyl)-propoxy]-4 methoxyphenethyl]-1H-imidazole hydrochloride, 10-50 microM) dose dependently abolished the response to bradykinin, while this response was only partially attenuated by nifedipine (10-50 microM; a voltage-operated Ca(2+) channel antagonist). Thapsigargin (an inhibitor of the sarcoplasmic reticulum calcium ATP ase pump, 1 microM) produced sustained contractions of guinea-pig gallbladder strips that were dependent on extracellular Ca(2+). After incubation of strips in a Ca(2+)-free medium with thapsigargin, replacement of Ca(2+) caused a large sustained contraction. We conclude that the contractile response of guinea-pig gallbladder to bradykinin is modulated by prostaglandin E(2). Bradykinin induced contractions of guinea-pig gallbladder are highly dependent on extracellular Ca(2+) which enters through store-operated Ca(2+) channels and partially through voltage-operated Ca(2+) channels. PMID- 11728433 TI - Effects of antagonism of NMDA receptors on transient lower esophageal sphincter relaxations in the dog. AB - Transient lower esophageal sphincter relaxation is the major cause of gastroesophageal reflux. Mechanisms underlying transient lower esophageal sphincter relaxation are poorly understood although gastric mechanosensitive vagal afferent pathways play a central role. Glutamate is a key transmitter of vagal afferents acting partly on NMDA receptors. The aim of this work was to study the effects on transient lower esophageal sphincter relaxation in awake dogs (n=5) of the competitive NMDA receptor antagonist cis-4-phosphonomethyl-2 piperidine carboxylic acid (CGS 19755; 0.3 and 3 mg/kg i.v., the high dose was given at two separate occasions to each dog). Transient lower esophageal sphincter relaxations were evoked by intragastric infusion of a liquid meal followed by air insufflation and were scored during a 45-min period. Neither dose produced any significant effect on the group average number of transient lower esophageal sphincter relaxations. Synchronous contractions of the esophagus were commonly seen during transient lower esophageal sphincter relaxation and CGS 19755 at both doses greatly reduced their occurrence. The findings indicate that NMDA receptor antagonism selectively inhibits the esophageal component of transient lower esophageal sphincter relaxation although the rate of transient lower esophageal sphincter relaxations is not consistently affected. PMID- 11728434 TI - PACAP-(6-38) inhibits the effects of vasoactive intestinal polypeptide, but not PACAP, on the small intestinal circular muscle. AB - Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating peptide-(1-38) (PACAP) have been found to stimulate distension-induced peristaltic motility in the guinea-pig isolated small intestine. In this study, we tested whether the putative VIP/PACAP receptor antagonist PACAP-(6-38) counteracts the properistaltic effect of VIP and PACAP in isolated segments of the guinea-pig small intestine. VIP (100 nM) and PACAP (30 nM) had a stimulatory effect, i.e., lowered the peristaltic pressure threshold at which peristaltic waves were triggered and enhanced the frequency of peristaltic waves. PACAP-(6 38) (3 microM) was per se without effect on peristalsis but prevented or reversed the peristaltic motor stimulation caused by VIP, when it was given before or after the agonist, respectively. PACAP-(6-38), however, failed to antagonize the properistaltic effect of PACAP. In ileal circular strips treated with tetrodotoxin (1 microM) and indomethacin (3 microM), spontaneous myogenic activity was inhibited by VIP (5-30 nM). This effect was significantly reduced by a pretreatment with PACAP-(6-38) (3 microM). A similar inhibition by PACAP-(1-38) (10-500 nM) was not influenced by the antagonist. It is concluded that PACAP-(6 38) is a VIP receptor antagonist, both in the peristaltic motor pathways and at the level of the circular muscle of the guinea-pig small intestine. The lack of a motor effect of PACAP-(6-38) on its own indicates that VIP acting on PACAP-(6-38) sensitive receptors (located on neurons and/or the smooth muscle) is unlikely to participate in peristaltic motor regulation. PMID- 11728435 TI - Hyperbaric oxygen treatment reduces carrageenan-induced acute inflammation in rats. AB - The present study was designed to assess the anti-inflammatory activity of hyperbaric oxygen treatment by comparing it with that of diclofenac, a nonsteroidal anti-inflammatory drug, and also to investigate whether hyperbaric oxygen treatment enhances the anti-inflammatory effect of diclofenac in carrageenan-induced paw edema which is commonly employed as an acute inflammation model in rats. Hyperbaric oxygen treatment and diclofenac (20 mg/kg) markedly reduced the carrageenan-induced paw edema in rats. In other words, they displayed anti-inflammatory activity. On the other hand, hyperbaric oxygen treatment did not consistently modify the anti-inflammatory effect of diclofenac in this model. PMID- 11728436 TI - STAM/EAST/Hbp adapter proteins--integrators of signalling pathways. AB - STAM/EAST/Hbp family of proteins consists of eight members well conserved from yeast to mammals. The basic domain architecture is comprised of an N-terminal Vps27, Hrs and STAM homology domain, a ubiquitin-interacting motif and a central Src homology-3 domain. Vertebrate members also carry an immunoreceptor tyrosine based activation motif. STAM/EAST/Hbp proteins become tyrosine-phosphorylated by a variety of cytokines and growth factors. STAM 1 and STAM 2A are involved in cytokine-mediated signalling for DNA synthesis and c-myc induction. EAST and STAM 2A/Hbp play a role in receptor-mediated endo- and exocytosis and probably also in the regulation of actin cytoskeleton. Knockout experiments implicate a role for STAM 1 in neural cell survival. A picture is emerging of STAM/EAST/Hbp proteins acting as integrators of thus far mechanistically disparate cellular signalling events. PMID- 11728437 TI - A look into kinesin's powerhouse. AB - Kinesins are microtubule-dependent motors that serve a multitude of cellular purposes. The conserved motor domain provides the energy required for these processes. Shortly after the solution of the first kinesin motor domain crystal structures the similarity to myosin and G-proteins was noted. By analogy, it was suspected that regions flanking the gamma-phosphate group of the nucleotide (in particular the so-called switch I and II regions) play important roles in the catalytic mechanism and the communication between the nucleotide cleft and the microtubule binding site. Since then, mutational analyses have supported this notion. Moreover, additional high-resolution structures have demonstrated that the switch regions can assume variable conformations. In one case, a comparison of an ADP state and an ATP-like state indicates a crucial involvement of the helix flanking switch II in modulating microtubule affinity. High-resolution structures of a kinesin-related protein mutated in the switch regions confirm the correlation between structural features in the switch vicinity and coupling of microtubule binding and nucleotide state. PMID- 11728439 TI - Identification of a structural motif of 23S rRNA interacting with 5S rRNA. AB - To identify RNA motifs interacting with 5S rRNA, a systematic evolution of ligands by exponential enrichment experiment was applied. Some of the resulting RNA aptamers contained a consensus sequence similar to the sequence in the loop region of helix 89 of 23S rRNA. We show that the synthetic helix 89 RNA motif indeed interacted with 5S rRNA and that the region around loop B of 5S rRNA was involved in this interaction. These results suggest the presence of a novel RNA RNA interaction between 23S rRNA and 5S rRNA which may play an important role in the ribosome function. PMID- 11728438 TI - Myosin VI, a new force in clathrin mediated endocytosis. AB - The integrity of the actin cytoskeleton and associated motor proteins are essential for the efficient functioning of clathrin mediated endocytosis at least in polarised cells. Myosin VI, the only motor protein so far identified that moves towards the minus end of actin filaments, is the first motor protein to be shown to associate with clathrin coated pits/vesicles at the plasma membrane and to modulate clathrin mediated endocytosis. Recent kinetic studies suggest that myosin VI may move processively along actin filaments providing clues about its functions in the cell. The possible role(s) of myosin VI in the sequential steps involved in receptor mediated endocytosis are discussed. PMID- 11728440 TI - The MED-7 transcriptional mediator encoded by let-49 is required for gonad and germ cell development in Caenorhabditis elegans. AB - Transcription mediators are evolutionarily conserved from yeast to human. We previously reported the specific in vivo roles of mediators during development. Transcriptional mediators including med-6, med-7, and med-10 were shown to be involved in the regulated transcription of specific genes, but not in the transcription of ubiquitous genes. In this report we have identified and characterized the Caenorhabditis elegans med-7 gene. A genetic mutation in the med-7 gene was identified by comparing genetic and physical maps and determining the molecular lesion. let-49 was found to have a nonsense mutation in the coding region of the med-7 gene. The identification of let-49 as the med-7 gene was confirmed by rescue experiments. The phenotype of the let-49 mutation indicated that the med-7 gene is required for normal postembryonic development. RNAi experiments showed that med-7 is also involved in embryogenesis and the gonad and germ cell development. PMID- 11728441 TI - An in vitro DNA virus for in vitro protein evolution. AB - In vitro virus is a molecular construct for in vitro protein evolution, which requires some mechanism to link phenotype to genotype. The first in vitro virus was realized by bonding a nascent protein with its coding mRNA via puromycin in in vitro translation. We report a new construct of in vitro DNA virus. The virion was a covalent cDNA-protein fusion, and virion formation did not require any modification of mRNA. Due to intactness of mRNA, this type of in vitro DNA virus will take the next step toward in vitro autonomous evolution, just like in vivo viral evolution in a cellstat. PMID- 11728442 TI - The serine protease inhibitor antithrombin III inhibits LPS-mediated NF-kappaB activation by TLR-4. AB - In Drosophila, the Toll family of proteins mediates the innate immune response. Toll is activated by Spaetzle, which is generated in response to pathogens via a serine protease cascade. We wished to investigate if lipopolysaccharides (LPS) might activate Toll-like receptor (TLR) 4 via a serine protease in humans. The serpin antithrombin III (ATIII) and the thrombin inhibitor hirudin both inhibited nuclear factor (NF)-kappaB activation by LPS and Lipid A. ATIII and hirudin were also able to inhibit LPS-induced NF-kappaB activation in cells stably transfected with TLR4. These results suggest that LPS may activate a mammalian serine protease, which generates a product required for TLR4 signalling. PMID- 11728443 TI - Conformational and functional significance of residue proline 17 in chicken muscle adenylate kinase. AB - The effect of mutation proline 17 on the multiple conformations and catalytic function in chicken muscle adenylate kinase (AK) has been studied. The substitution of proline 17 with glycine or valine altered the distribution of multiple conformations. Compared with the wild-type enzyme, the P17G and P17V mutants contained decreased fraction of minor conformer from 18% to 9% and 11%, respectively. Due to the mutation, the enzyme showed lower secondary structural content, poorer affinity to substrates or substrate analogues, and reduced catalytic efficiency. The results revealed the significance of proline 17 in the conformation and function of AK. PMID- 11728444 TI - Spermidine is essential for normal proliferation of trypanosomatid protozoa. AB - Trypanosomatid parasites containing a metabolically unstable ornithine decarboxylase (ODC) are naturally resistant to high levels of alpha difluoromethylornithine (DFMO) because this ODC inhibitor, though causing a drastic reduction of intracellular putrescine, elicits only a moderate decrease of the spermidine endogenous pool. In this study we have used a combination of DFMO with cyclohexylamine (CHA; bis-cyclohexylammonium sulfate), an inhibitor of spermidine synthase, to reach a more complete depletion of spermidine. Under these conditions we have observed the arrest of proliferation not only in trypanosomatids with stable ODC but also in parasites with an enzyme of high turnover rate. In all cases the reinitiation of proliferation occurred only after the addition of exogenous spermidine, and neither putrescine nor spermine were able to induce the same effect. PMID- 11728445 TI - Anandamide amidohydrolase activity, released in the medium by Tetrahymena pyriformis. Identification and partial characterization. AB - Anandamide, an endogenous cannabinoid receptor ligand, was rapidly metabolized by Tetrahymena pyriformis in vivo. Metabolic products were mainly phospholipids as well as neutral lipids, including small amounts of free arachidonic acid. Anandamide amidohydrolase activity was detected in the culture medium by the release of [3H]arachidonic acid from [3H]anandamide, in a time- and concentration dependent manner. Kinetic experiments demonstrated that the released enzyme had an apparent K(m) of 3.7 microM and V(max) 278 pmol/min/mg protein. Amidohydrolase activity was maximal at pH 9-10, was abolished by phenylmethylsulfonyl fluoride and was Ca(2+)- and Mg(2+)-independent. Thus, T. pyriformis is capable of hydrolyzing anandamide in vivo and releasing amidohydrolase activity. PMID- 11728446 TI - Mechanism of Kex2p inhibition by its proregion. AB - Many proteases are produced as zymogens bearing an N-terminal proregion acting both as intramolecular chaperone and as enzyme inhibitor. We studied here the inhibition mechanism of the yeast proprotein convertase Kex2p by its proregion. A recombinant secreted and soluble form of Kex2p was produced in Pichia pastoris and its enzymatic properties toward a fluorogenic synthetic peptide were characterized. Recombinant Escherichia coli-produced Kex2p proregion specifically and potently inhibited the enzyme, with an IC(50) of 160 nM. Exploration of the inhibition mechanism revealed that the proregion behaved as a mixed inhibitor. PMID- 11728447 TI - Gene silencing by S-adenosylmethionine in muscle differentiation. AB - A well-characterised experimental system, the myogenin gene in C2C12 muscle cell culture, was chosen to better understand the methylation mechanism underlying the regulation of gene expression. We already demonstrated that demethylation dynamics of a specific CpG site in the 5'-flanking region of myogenin well correlates with gene expression and terminal differentiation. Here we demonstrate that S-adenosylmethionine-sulphate-p-toluenesulphonate (SAM) inhibits myogenin expression and myoblast differentiation by delaying the demethylation of specific CpG in differentiating myoblasts. These results suggest new perspectives in methylation mechanisms and the use of SAM in the partial silencing of gene expression, as it could be required in disease treatment. PMID- 11728448 TI - Dioxin induces a novel nuclear factor, DIF-3, that is implicated in spermatogenesis. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD; dioxin), a member of a class of environmental pollutants represented by polychlorinated dibenzo-p-dioxins and dibenzofurans, is one of the most toxic artificial compounds ever developed. In this study, we identified a novel TCDD target gene, DIF-3 (dioxin inducible factor-3), by cDNA representational difference analysis. DIF-3 protein is a nuclear factor and possesses a zinc-finger motif at its N-terminus. High DIF-3 mRNA expression in the testes was demonstrated by Northern blot analysis and abundant DIF-3 protein was detected during spermatogenesis. Thus, these results suggest that DIF-3 may be a target gene mediating the reproductive toxicity induced by TCDD. PMID- 11728449 TI - Histamine increases the expression of LOX-1 via H2 receptor in human monocytic THP-1 cells. AB - Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a member of the scavenger receptor family, and is known to be expressed in monocytes/macrophages. We investigated the effect of histamine on the expression of LOX-1 in cells of the human monocytic leukemia cell line THP-1. Histamine as well as forskolin and dibutyryl cyclic AMP (Bt2-cAMP) stimulated the THP-1 monocytes to express the LOX-1 gene at the transcription level. This histamine effect on LOX-1 gene expression, via the histamine H2 receptor-mediated cAMP signal transduction pathway, was reduced after differentiation of the cells into macrophages, even though forskolin and Bt2-cAMP still enhanced the gene expression. The alteration of the responsiveness of LOX-1 expression to histamine was related to suppressed expression of the H2 receptor in THP-1 macrophages. The switch of the predominant class of histamine receptors between H1 and H2 would modulate the effects of histamine on LOX-1 gene expression in monocytes and macrophages, and therefore, would play a certain role in the inflammatory aspects of atherogenesis. PMID- 11728450 TI - Functional and molecular characterization of a peptide transporter in the rat PC12 neuroendocrine cell line. AB - We have studied functional properties of peptide transport in the pheochromocytoma neuroendocrine cell line from rat. The neutral peptide D-Phe-L Ala (resistant to hydrolysis) is a good substrate for uptake into these cells. Transport is substantially inhibited by diethylpyrocarbonate pretreatment and is stimulated by external acidification. It is sodium-independent and, unexpectedly, insensitive to membrane potential. Peptide uptake is inhibited by a wide variety of other di- and tripeptides but not by amino acids. The neuropeptide kyotorphin (opioid dipeptide (L-Tyr-L-Arg)) inhibits uptake of labelled peptide and trans stimulates efflux showing that it is a transported substrate. These findings are discussed in relation to the molecular basis and physiological role of this transport system. PMID- 11728451 TI - Calcium-dependent cysteine reactivities in the neuronal calcium sensor guanylate cyclase-activating protein 1. AB - Guanylate cyclase-activating protein 1 (GCAP-1) is a Ca(2+)-sensing protein in vertebrate photoreceptor cells. It activates a membrane-bound guanylate cyclase. Three of four cysteines present in wild-type GCAP-1 were accessible to the thiol modifying reagent 5,5'-dithio-bis-(2-nitrobenzoic acid) in the presence of Ca(2+). Only Cys106 became exposed to the solvent after Ca(2+)-chelation. Since Cys106 is located in EF-hand 3, we could determine an apparent K(D) of 2.9 microM for Ca(2+) binding to this site with a fast off-rate (t approximately 2 ms). We conclude that the rapid dissociation of Ca(2+) from EF-hand 3 in GCAP-1 triggers activation of guanylate cyclase in rod cells. PMID- 11728452 TI - NF-kappaB is required for TNF-alpha-directed smooth muscle cell migration. AB - Migration of vascular smooth muscle cells (VSMC) is a crucial event in the formation of vascular stenotic lesions. Tumor necrosis factor-alpha (TNF-alpha) is elaborated by VSMC in atherosclerosis and following angioplasty. We investigated the role of nuclear factor-kappaB (NF-kappaB) in human VSMC migration induced by TNF-alpha. Adenoviral expression of a mutant form of the inhibitor of NF-kappaB, IkappaB-alphaM, suppressed TNF-alpha-triggered degradation of cellular IkappaB-alpha, inhibited activation of NF-kappaB, and attenuated TNF-alpha-induced migration. Further, IkappaB-alphaM suppressed TNF alpha-stimulated release of interleukin-6 and -8 (IL-6 and IL-8). Neutralization of IL-6 and IL-8 with appropriate antibodies reduced TNF-alpha-induced VSMC migration. Addition of recombinant IL-6 and IL-8 stimulated migration. Collectively, our data provide initial evidence that TNF-alpha-mediated VSMC migration requires NF-kappaB activation and is associated with induction of IL-6 and IL-8 which act in an autocrine manner. PMID- 11728453 TI - Non-steroidal anti-inflammatory drugs inhibit matrix metalloproteinase-2 expression via repression of transcription in lung cancer cells. AB - Recent studies show that up-regulation of cyclooxygenase-2 (COX-2) in human cancer cells induces activation of matrix metalloproteinases (MMPs) and increase of metastatic potential. In this study, we investigate the effect of a COX-2 selective inhibitor, NS398, on the expression and enzymatic activity of MMPs in human lung cancer cells. We found that NS398 inhibited MMP-2, not MMP-9, mRNA expression. NS398 also reduced the amount of MMP-2, not MMP-9, released into the medium. Additionally, this COX-2 inhibitor attenuated the degrading activity of MMP-2 as demonstrated by gelatin zymography. Investigation of cellular MMP-2 by Western blotting indicated that synthesis and processing of MMP-2 was significantly suppressed by NS398. We performed promoter activity assay to address whether NS398 might affect MMP-2 gene transcription. Our results indicated that NS398 directly inhibited MMP-2 promoter activity. However, the inhibitory effect of NS398 is not fully dependent on inhibition of COX-2 because a high concentration of NS398 was needed to suppress MMP-2 expression and addition of prostaglandin E2 only partially reversed the action of NS398. Moreover, a non-selective COX inhibitor indomethacin also suppressed the expression of MMP-2. Taken together, these results indicate that non-steroidal anti-inflammatory drugs suppress MMP-2 expression via repression of transcription and support the notion that COX inhibitors may be useful in inhibition and/or prevention of metastasis. PMID- 11728454 TI - Anti-apoptotic protein survivin plays a significant role in tubular morphogenesis of human coronary arteriolar endothelial cells by hypoxic preconditioning. AB - Brief exposure of endothelial cells to oxidative stress induced by hypoxia followed by reoxygenation enhances tube formation. Our study provides evidence that hypoxic preconditioning accelerates tubular morphogenesis along with the activation of reactive oxygen species-inducible nuclear transcription factor kappaB (NF-kappaB), phosphatidylinositol 3-kinase (PI3-kinase) and broad-spectrum anti-apoptotic protein survivin in human coronary arteriolar endothelial cells (HCAEC). The formation of tubular morphogenesis was inhibited by using the PI3 kinase and NF-kappaB antagonists LY294002 and SN50 respectively. The activation of survivin by hypoxic preconditioning was also inhibited by LY294002 and SN50 along with increased apoptosis in HCAEC. These data demonstrate a crucial role of PI3-kinase/Akt/NF-kappaB/survivin signaling in tubular morphogenesis of HCAEC triggered by hypoxic preconditioning. PMID- 11728455 TI - The Arabidopsis thaliana ethylene-responsive element binding protein (AtEBP) can function as a dominant suppressor of Bax-induced cell death of yeast. AB - We identified genes based on screening of an Arabidopsis cDNA library for functional suppressors of mouse Bax-induced cell death of yeast cells. Interestingly, the cDNA encoding AtEBP, known as Arabidopsis thaliana ethylene responsive element binding protein, was isolated numerous times in the functional screen (82% of all suppressors). Full-length AtEBP and its localization to the nucleus were essential for the suppression of Bax-induced cell death. Morphological abnormality of intracellular network that is a hallmark of Bax induced cell death was attenuated by expression of AtEBP. PMID- 11728456 TI - Urokinase-type plasminogen activator up-regulates the expression of its cellular receptor through a post-transcriptional mechanism. AB - We have recently reported that the urokinase-type plasminogen activator (uPA) up regulates the cell surface expression of its own receptor (uPAR) in several cell types, independently of its enzymatic activity. uPA has no effect on kidney 293 cells which do not express uPAR and then cannot bind uPA. Kidney cells, transfected with the coding region of uPAR cDNA, express very large amounts of uPAR and respond to uPA stimulation by regulating uPAR both at mRNA and protein levels. uPA effect occurs also in the presence of the transcriptional inhibitor dichloro-ribobenzimidazole, whereas it is abolished by the protein synthesis inhibitor cycloheximide. Moreover, uPA-dependent uPAR up-regulation correlates with the increase of a complex between the coding region of uPAR mRNA and an unknown cellular factor. We then propose that uPA regulates uPAR expression at a post-transcriptional level, by promoting the binding of uPAR mRNA to a stabilizing factor. PMID- 11728457 TI - Direct interaction between a membrane domain subunit and a connector subunit in the H(+)-translocating NADH-quinone oxidoreductase. AB - When Paracoccus denitrificans membranes were treated with a crosslinker, m maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), a cross-linked product of M(r) approximately 31 kDa was found which reacted with antibodies against the hydrophobic subunit Nqo7 and the connector subunit Nqo6. NaI treatment of the Paracoccus membranes before, but not after, the crosslinking step prevented the formation of the 31 kDa band. When Nqo7 and Nqo6 were coexpressed in Escherichia coli, both subunits were located in the membrane fraction. MBS treatment of the E. coli membranes generated the 31 kDa band as in the Paracoccus membranes. These results indicate that Nqo7 interacts with probable N2-binding Nqo6. PMID- 11728458 TI - The antimicrobial peptides lactoferricin B and magainin 2 cross over the bacterial cytoplasmic membrane and reside in the cytoplasm. AB - The localization of immunolabelled antimicrobial peptides was studied using transmission electron microscopy. Staphylococcus aureus and Escherichia coli were exposed to lactoferricin B (17-41), lactoferricin B (17-31) and D-lactoferricin B (17-31). E. coli was also exposed to cecropin P1 and magainin 2. The lactoferricins were found in the cytoplasm of both bacteria. In S. aureus the amount of cytoplasmic lactoferricin B (17-41) was time- and concentration dependent, reaching a maximum within 30 min. Cecropin P1 was confined to the cell wall, while magainin 2 was found in the cytoplasm of E. coli. The finding of intracellularly localized magainin is not reported previously. PMID- 11728459 TI - Drosophila Chk2 is required for DNA damage-mediated cell cycle arrest and apoptosis. AB - Chk2 is a major target of ataxia telangiectasia-mutated (ATM) and ATM- and Rad3 related (ATR). Germline mutations in Chk2 have been identified in a subset of patients with Li-Fraumeni syndrome, suggesting that Chk2 is a tumor suppressor gene. To investigate the role of Chk2 in multicellular organisms, a Drosophila chk2 (Dmchk2) mutant was generated. Dmchk2 mutants are viable but show defects in maintaining genome stability and are highly sensitive to ionizing radiation. Interestingly, mutating Dmchk2 completely blocks DNA damage-induced apoptosis and partially blocks DNA damage-induced cell cycle arrest. These results indicate that Chk2 protein plays a crucial role in the DNA damage response pathway mediating cell cycle arrest and apoptosis, and that the ATM-Chk2 pathway is likely conserved in Drosophila. PMID- 11728460 TI - Thr90 is a key residue of the bacteriorhodopsin proton pumping mechanism. AB - Mutation of Thr90 to Ala has a profound effect on bacteriorhodopsin properties. T90A shows about 20% of the proton pumping efficiency of wild type, once reconstituted into liposomes. Mutation of Thr90 influences greatly the Schiff base/Asp85 environment, as demonstrated by altered lambda(max) of 555 nm and pK(a) of Asp85 (about 1.3 pH units higher than wild type). Hydroxylamine accessibility is increased in both dark and light and differential scanning calorimetry and visible spectrophotometry show decreased thermal stability. These results suggest that Thr90 has an important structural role in both the unphotolysed bacteriorhodopsin and in the proton pumping mechanism. PMID- 11728461 TI - Homocysteine attenuates endothelial haem oxygenase-1 induction by nitric oxide (NO) and hypoxia. AB - The disrupted metabolism of homocysteine (Hcy) causes hyperhomocysteinemia, a condition associated with the impairment of nitric oxide (NO) bio-availability, tissue hypoxia and increased risk of vascular disease. Here, we examined how Hcy modulates the induction of the stress protein haem oxygenase-1 (HO-1) evoked by NO releasing agents and hypoxia in vascular endothelial cells. We found that Hcy (0.5 mM) markedly reduced the increase in haem oxygenase activity and HO-1 protein expression induced by sodium nitroprusside (SNP, 0.5 mM) but did not affect HO-1 activation mediated by S-nitroso-N-acetyl-penicillamine. Cells pre treated with Hcy followed by addition of fresh medium containing SNP still exhibited an augmented haem oxygenase activity. Interestingly, high levels of Hcy were also able to abolish hypoxia-mediated HO-1 expression in a concentration dependent manner. These novel findings indicate that hyperhomocysteinemia interferes with crucial signaling pathways required by cells to respond and adapt to stressful conditions. PMID- 11728463 TI - Transformation of acridone synthase to chalcone synthase. AB - Acridone synthase (ACS) and chalcone synthase (CHS) catalyse the pivotal reactions in the formation of acridone alkaloids or flavonoids. While acridone alkaloids are confined almost exclusively to the Rutaceae, flavonoids occur abundantly in all seed-bearing plants. ACSs and CHSs had been cloned from Ruta graveolens and shown to be closely related polyketide synthases which use N methylanthraniloyl-CoA and 4-coumaroyl-CoA, respectively, as the starter substrate to produce the acridone or naringenin chalcone. As proposed for the related 2-pyrone synthase from Gerbera, the differential substrate specificities of ACS and CHS might be attributed to the relative volume of the active site cavities. The primary sequences as well as the immunological cross reactivities and molecular modeling studies suggested an almost identical spatial structure for ACS and CHS. Based on the Ruta ACS2 model the residues Ser132, Ala133 and Val265 were assumed to play a critical role in substrate specificity. Exchange of a single amino acid (Val265Phe) reduced the catalytic activity by about 75% but grossly shifted the specificity towards CHS activity, and site-directed mutagenesis replacing all three residues by the corresponding amino acids present in CHS (Ser132Thr, Ala133Ser and Val265Phe) fully transformed the enzyme to a functional CHS with comparatively marginal ACS activity. The results suggested that ACS divergently has evolved from CHS by very few amino acid exchanges, and it remains to be established why this route of functional diversity has developed in the Rutaceae only. PMID- 11728462 TI - Antigen-independent selection of stable intracellular single-chain antibodies. AB - The intracellular expression of single-chain Fv antibody fragments (scFv) in eukaryotic cells has an enormous potential in functional genomics and therapeutics [Marasco (1997) Gene Ther. 4, 11-15; Richardson and Marasco (1995) Trends Biotechnol. 13, 306-310]. However, the application of these so-called intrabodies is currently limited by their unpredictable behavior under the reducing conditions encountered inside eukaryotic cells, which can affect their stability and solubility properties [Worn et al. (2000) J. Biol. Chem. 275, 2795 2803; Biocca et al. (1995) Bio/Technology 13, 1110-1115]. We present a novel system that enables selection of stable and soluble intrabody frameworks in vivo without the requirement or knowledge of antigens. This system is based on the expression of single-chain antibodies fused to a selectable marker that can control gene expression and cell growth. Our results show that the activity of a selectable marker fused to well characterized scFvs [Worn et al. (2000) J. Biol. Chem. 275, 2795-2803] correlates with the solubility and stability of the scFv moieties. This method provides a unique tool to identify stable and soluble scFv frameworks, which subsequently serve as acceptor backbones to construct intrabody complementarity determining region libraries by randomization of hypervariable loops. PMID- 11728464 TI - CD30L up-regulates CD30 and IL-4 expression by T cells. AB - CD30L is frequently expressed on acute myeloid leukemia (AML) blasts. Its presence is associated with the co-expression of interleukin-4 (IL-4) receptor and with the expansion of specific T-helper 2 (Th2) cell subsets producing IL-4 and expressing CD30. Recombinant CD30L-bearing cells up-regulated the expression of surface CD30 and increased the production of IL-4 and soluble (s) CD30 by co cultured T cells. These findings were confirmed with AML blasts expressing surface CD30L, where blocking anti-CD30 antibodies completely abolished the release of sCD30 and reduced the production of IL-4. Our data indicates a direct role of CD30L(+) neoplastic cells in driving the immune response toward a Th2 polarized non-protective state. PMID- 11728465 TI - Blepharismins, produced by the protozoan, Blepharisma japonicum, form ion permeable channels in planar lipid bilayer membranes. AB - Blepharismins are polycyclic quinones found in the pigment granules of the ciliated protozoan, Blepharisma. Exposure to purified blepharismins results in lethal damage to several other ciliates. We here report that, at cytotoxic concentrations, blepharismins formed cation-selective channels in planar phospholipid bilayer membranes. The channels formed in a diphytanoylphosphatidylcholine bilayer had a K(+)/Cl(-) permeability ratio of 6.6:1. Single channel recordings revealed the conductance to be quite heterogeneous, ranging from 0.2 to 2.8 nS in solutions containing 0.1 M KCl, possibly reflecting different states of aggregation of blepharismin. Our observations suggest that channel formation is a cytotoxic mechanism of blepharismin's action against predatory protozoa. PMID- 11728466 TI - Two parameters improve efficiency of mitochondrial uptake of adenylate kinase: decreased folding velocity and increased propensity of N-terminal alpha-helix formation. AB - The long isoform of eukaryotic adenylate kinase has a dual subcellular location in the cytoplasm and in the mitochondrial intermembrane space. Protein sequences and modifications are identical in both locations. In yeast, the bulk of the major form of adenylate kinase (Aky2p) is in the cytoplasm and, in the steady state, only 5-8% is sorted to the mitochondrial intermembrane space. Since the reasons for exclusion from mitochondrial import are unclear, we have constructed aky2 mutants with elevated mitochondrial uptake efficiency of Aky2p in vivo and in vitro. We have analyzed the effect of the mutations on secondary structure prediction in silico and have tested folding velocity and folding stability. One type of mutants displayed decreased proteolytic stability and retarded renaturation kinetics after chaotropic denaturation implying that deterioration of folding leads to prolonged presentation of target information to mitochondrial import receptors, thereby effecting improved uptake. In a second type of mutants, increased import efficiency was correlated with an increased probability of formation of an alpha-helix with increased amphipathic moment at the N-terminus suggesting that targeting interactions with mitochondrial import receptors had been improved at the level of binding affinity. PMID- 11728467 TI - S-Adenosyl-L-homocysteine hydrolase is sequestered into actin rods in Dictyostelium discoideum spores. AB - Here we show evidence that S-adenosyl-L-homocysteine hydrolase (SAHH) is linked to the actin cytoskeleton. Actin rods formed in Dictyostelium discoideum spores during the final stage of development are structurally composed of novel bundles of actin filaments. SAHH only accumulates with actin at this stage of development in the life cycle of D. discoideum. Recently SAHH is believed to be a target for antiviral chemotherapy and the suppression of T cells. Our finding may contribute to designing novel antiviral and immunosuppressive drugs. PMID- 11728468 TI - Citrulline, a novel compatible solute in drought-tolerant wild watermelon leaves, is an efficient hydroxyl radical scavenger. AB - Drought-tolerant wild watermelon accumulates high levels of citrulline in the leaves in response to drought conditions. In this work, the hydroxyl radical scavenging activity of citrulline was investigated in vitro. The second-order rate constant for the reaction between citrulline and hydroxyl radicals was found to be 3.9x10(9) M(-1) s(-1), demonstrating that citrulline is one of the most efficient scavengers among compatible solutes examined so far. Moreover, citrulline effectively protected DNA and an enzyme from oxidative injuries. Liquid chromatography-mass spectrometry analysis revealed that at least four major products were formed by the reaction between citrulline and hydroxyl radicals. Activities of metabolic enzymes were not inhibited by up to 600 mM citrulline, indicating that citrulline does not interfere with cellular metabolism. We reasoned, from these results, that citrulline contributes to oxidative stress tolerance under drought conditions as a novel hydroxyl radical scavenger. PMID- 11728469 TI - Hydrogen peroxide is a regulator of ABI1, a protein phosphatase 2C from Arabidopsis. AB - Protein phosphatases 2C (PP2Cs) exhibit diverse regulatory functions in signalling pathways of animals, yeast and plants. ABI1 is a PP2C of Arabidopsis that exerts negative control on signalling of the phytohormone abscissic acid (ABA). Characterisation of the redox sensitivity of ABI1 revealed a strong enzymatic inactivation by hydrogen peroxide (H2O2) which has recently been implicated as a secondary messenger of ABA signalling. H2O2 reversibly inhibited ABI1 activity in vitro with an IC(50) of approximately 140 microM in the presence of physiological concentrations of glutathione. In addition, ABI1 was highly susceptible to inactivation by phenylarsine oxide (IC(50)=3-4 microM) indicative for the facile oxidation of vicinal cysteine residues. Thus, H2O2 generated during ABA signalling seems to inactivate the negative regulator of the ABA response. PMID- 11728470 TI - A novel dimer of a C-type lectin-like heterodimer from the venom of Calloselasma rhodostoma (Malayan pit viper). AB - We have isolated a potent platelet aggregation inducer from the crude venom of Calloselasma rhodostoma (Malayan pit viper), termed rhodoaggretin, with a novel oligomeric structure consisting of a dimer of C-type lectin-like heterodimers. On the basis of its native molecular mass of 66 kDa, and a M(r) of 30 kDa for its disulfide-linked alphabeta-heterodimer, we propose that rhodoaggretin exists as a (alphabeta)2 complex in the native state. We postulate that the di-dimer is stabilized by non-covalent interactions as well as by an intersubunit disulfide bridge between the two alphabeta-heterodimers. This conclusion is based on the following observations: (a) sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the non-reduced rhodoaggretin gave a major 28 and a minor 52 kDa band. (b) Prior treatment of rhodoaggretin with a limited amount of 2-mercaptoethanol (2-ME; 0.1%) resulted in the complete abolishment of the 52 kDa band in SDS-PAGE. (c) Two-dimensional SDS-PAGE in the presence of 3% 2-ME showed that both the 28 and 52 kDa bands gave two bands each with M(r)s of 18 (alpha subunit) and 15 (beta-subunit) kDa. (d) Mass spectrometric analyses showed that purified rhodoaggretin had a M(r) of 30155.39+/-3.25 Da while its s pyridylethylated alpha- and beta-subunits had M(r)s of 16535.62+/-2.98 and 15209.89+/-1.61 Da respectively. These molecular weight data suggested the presence of 15 cysteinyl residues in rhodoaggretin as compared to the 14 that are reported for the heterodimeric C-type lectin-like proteins. This extra cysteinyl residue is a candidate for the formation of the intersubunit disulfide bond in the (alphabeta)2 complex. (e) Homology structural modeling studies showed that the extra cysteinyl residue can indeed form a disulfide bond that covalently links the two alphabeta-heterodimers as proposed above. PMID- 11728471 TI - Determination of amino acid sequence responsible for suppression of bone resorption by serum calcium-decreasing factor (caldecrin). AB - We previously reported on the serum calcium-decreasing activity of recombinant protein factor referred to as caldecrin [Tomomura et al. (1995) J. Biol. Chem. 270, 30315-30321]. To address the mechanism of this serum calcium-decreasing activity, we investigated the effect of rat caldecrin on osteoclastic bone resorbing activity. Wild-type caldecrin suppressed resorption pit formation by osteoclast on a dentine slice in a dose-dependent manner. The suppressive effect on the bone resorption was not affected by treatment of caldecrin with phenylmethyl sulfonyl fluoride or by use of protease-deficient mutant caldecrins. Recombinant procaldecrin (-13-239), and its fragments (-13-125), (1-111), (1-46), (47-111), and (126-239) were expressed as His-tagged thioredoxin fusion proteins and investigated for their ability to suppress bone resorption. The proform (-13 239) and fragment (-13-125) did not affect the suppressive activity, whereas fragments (1-111) and (126-239) did suppress the bone resorption. The bone resorbing activity was also suppressed by fragment (47-111), not by fragment (1 46). Overlapping fragments (47-62), (47-79), (47-98), (56-111), (71-111), and (85 111) were compared for their suppressive activity. The fragments (47-62) and (85 111) did not affect the activity, but the other fragments suppressed the bone resorption. A synthetic peptide having the (71-79) sequence suppressed the bone resorption. These results suggest that amino acid sequence corresponding to rat caldecrin (aa 71-79) is responsible for the suppression of bone resorption by caldecrin. PMID- 11728473 TI - K(+) channel profile and electrical properties of Arabidopsis root hairs. AB - Ion channels and solute transporters in the plasma membrane of root hairs are proposed to control nutrient uptake, osmoregulation and polar growth. Here we analyzed the molecular components of potassium transport in Arabidopsis root hairs by combining K(+)-selective electrodes, reverse transcription-PCR, and patch-clamp measurements. The two inward rectifiers AKT1 and ATKC1 as well as the outward rectifier GORK dominated the root hair K(+) channel pool. Root hairs of AKT1 and ATKC1 loss-of-function plants completely lack the K(+) uptake channel or exhibited altered properties, respectively. Upon oligochitin-elicitor treatment of root hairs, transient changes in K(+) fluxes and membrane polarization were recorded in wild-type plants, while akt1-1 root hairs showed a reduced amplitude and pronounced delay in the potassium re-uptake process. This indicates that AKT1 and ATKC1 represent essential alpha-subunits of the inward rectifier. Green fluorescent protein (GFP) fluorescence following ballistic bombardment with GORK promoter-GFP constructs as well as analysis of promoter-GUS lines identified this K(+) outward rectifier as a novel ion channel expressed in root hairs. Based on the expression profile and the electrical properties of the root hair plasma membrane we conclude that AKT1-, ATKC- and GORK-mediated potassium transport is essential for osmoregulation and repolarization of the membrane potential in response to elicitors. PMID- 11728472 TI - Recombinant holophytochrome in Escherichia coli. AB - We have successfully co-expressed two genes from the bilin biosynthetic pathway of Synechocystis together with cyanobacterial phytochrome 1 (Cph1) from the same organism to produce holophytochrome in Escherichia coli. Heme oxygenase was used to convert host heme to biliverdin IXalpha which was then reduced to phycocyanobilin via phycocyanobilin:ferredoxin oxidoreductase, presumably with the aid of host ferredoxin. In this host environment Cph1 apophytochrome was able to autoassemble with the phycocyanobilin in vivo to form fully photoreversible holophytochrome. The system can be used as a tool for further genetic studies of phytochrome function and signal transduction as well as providing an excellent source of holophytochrome for physicochemical studies. PMID- 11728474 TI - Horizontal transfer of a bacterial gene involved in polyglutamate biosynthesis to the plant-parasitic nematode Meloidogyne artiellia. AB - Analysis of a genomic fragment from the plant parasitic nematode Meloidogyne artiellia revealed the presence of a gene which, in bacteria, is involved in the formation of polyglutamate capsule. Searching of various databases, including the Caenorhabditis elegans genome sequence and the large EST datasets from a variety of parasitic nematodes, showed that no similar genes have been identified in other nematodes or in any other eukaryotic organisms. The M. artiellia gene has a typical eukaryotic structure and its mRNA is present in the intestine. The gene is expressed in all life cycle stages tested. These findings demonstrate horizontal gene transfer may be important in catalyzing the diversification of nematode lineages. PMID- 11728475 TI - Mutational analysis of the iron binding site of Saccharomyces cerevisiae ferroxidase Fet3. An in vivo study. AB - The role of residues predicted to be involved in the binding of iron by the yeast ferroxidase Fet3 has been studied by site-directed mutagenesis. The effect of Fet3 mutations E185A, E185Q, Y354F, D409V and H489D has been investigated in vivo by kinetic analyses of high affinity iron uptake. Our results indicate that Glu 185 is critical for the binding of iron, since substitution of this residue with Ala or Gln strongly affects both growth and the kinetic parameters of high affinity iron uptake, greatly increasing K(m). Mutations Y354F and D409V result in less severe alteration of high affinity iron uptake, while mutant H489D is unable to grow under conditions of iron limitation. PMID- 11728476 TI - Vacuolation induced by VacA toxin of Helicobacter pylori requires the intracellular accumulation of membrane permeant bases, Cl(-) and water. AB - The protein vacuolating toxin A (VacA) of Helicobacter pylori converts late endosomes into large vacuoles in the presence of permeant bases. Here it is shown that this phenomenon corresponds to an accumulation of permeant bases and Cl(-) in HeLa cells and requires the presence of extracellular Cl(-). The net influx of Cl(-) is due to electroneutral, Na(+), K(+), 2Cl(-) cotransporter-mediated transport. Cell vacuolation leads to cell volume increase, consistent with water flux into the cell, while hyper-osmotic media decreased vacuole formation. These data represent the first evidence that VacA-treated cells undergo an osmotic unbalance, reinforcing the hypothesis that the VacA chloride channel is responsible for cell vacuolation. PMID- 11728477 TI - Production of beta-defensin-2 by human colonic epithelial cells induced by Salmonella enteritidis flagella filament structural protein. AB - We recently showed that FliC of Salmonella enteritidis increased human beta defensin-2 (hBD-2) expression, and now describe the signaling responsible pathway. FliC increased the intracellular Ca(2+) concentration ([Ca(2+)](in)) in Caco-2 cells. The [Ca(2+)](in) increase induced by FliC was prevented by U73122 and heparin, but not by chelating extracellular Ca(2+) or pertussis toxin. The FliC-induced increase in hBD-2 promoter activity via nuclear factor kappaB (NF kappaB) was also inhibited by chelation of intracellular Ca(2+) or by U73122. We conclude that FliC increased [Ca(2+)](in) via inositol 1,4,5-trisphosphate, which was followed by up-regulating hBD-2 mRNA expression via an NF-kappaB-dependent pathway. PMID- 11728481 TI - Gestational diabetes mellitus: prevalence, risk factors, maternal and infant outcomes. AB - OBJECTIVES: To study prevalence, risk factors, and maternal and infant outcomes of women with gestational diabetes mellitus (GDM). METHODS: A retrospective cohort study was performed based on 111563 pregnancies delivered between 1991 through 1997 in 39 hospitals in northern and central Alberta, Canada. Multivariate logistic regression was used to estimate the odds ratios with 95% confidence intervals, and to control for confounding variables. RESULTS: The prevalence of GDM was 2.5%. Risk factors for GDM included age >35 years, obesity, history of prior neonatal death, and prior cesarean section. Teenage mothers and women who drank alcohol were less likely to have GDM. Mothers with GDM were at increased risk of presenting with pre-eclampsia, premature rupture of membranes, cesarean section, and preterm delivery. Infants born to mothers with GDM were at higher risk of being macrosomic or large-for-gestational-age. CONCLUSIONS: Specific conditions predispose to GDM which itself is associated with a significantly increased risk of maternal and fetal morbidity. PMID- 11728478 TI - Selenium spares ascorbate and alpha-tocopherol in cultured liver cell lines under oxidant stress. AB - The selenoenzyme thioredoxin reductase (TR) can recycle ascorbic acid, which in turn can recycle alpha-tocopherol. Therefore, we evaluated the role of selenium in ascorbic acid recycling and in protection against oxidant-induced loss of alpha-tocopherol in cultured liver cells. Treatment of HepG2 or H4IIE cultured liver cells for 48 h with sodium selenite (0-116 nmol/l) tripled the activity of the selenoenzyme TR, measured as aurothioglucose-sensitive dehydroascorbic acid (DHA) reduction. However, selenium did not increase the ability of H4IIE cells to take up and reduce 2 mM DHA, despite a 25% increase in ascorbate-dependent ferricyanide reduction (which reflects cellular ascorbate recycling). Nonetheless, selenium supplements both spared ascorbate in overnight cultures of H4IIE cells, and prevented loss of cellular alpha-tocopherol in response to an oxidant stress induced by either ferricyanide or diazobenzene sulfonate. Whereas TR contributes little to ascorbate recycling in H4IIE cells, selenium spares ascorbate in culture and alpha-tocopherol in response to an oxidant stress. PMID- 11728482 TI - Beta-human chorionic gonadotropin and prolactin assays in cervicovaginal secretions as a predictor of preterm delivery. AB - OBJECTIVE: To evaluate and to compare the predictive value of cervicovaginal beta hCG and prolactin levels in spontaneous preterm delivery. METHODS: The preterm labor group and normal pregnancy group consisted of 17 and 43 patients between 24 and 36 weeks' gestation, respectively. A single cervicovaginal beta-hCG and prolactin measurement were made in both groups. RESULTS: Cervicovaginal beta-hCG and prolactin levels were significantly higher in the preterm group when compared with those of the term delivery group (P=0.031, P=0.026, respectively). The optimal cut-off value for beta-hCG (27.1 mIU/ml) gave a sensitivity level of 87.5% (47.4-97.9; 95% C.I.) at a specificity of 65.4% (50.9-78.0; 95% C.I.) with positive and negative predictive values of 28% and 97%, respectively. The optimal cut-off value for prolactin (1.8 ng/ml) gave a sensitivity level of 50% (16.0 84.0; 95% C.I.) at a specificity of 96% (86.8-99.4; 95% C.I.) with positive and negative predictive values of 67% and 93%, respectively. CONCLUSIONS: Cervicovaginal beta-hCG measurement in patients with preterm labor may be used as a predictive test. Cervicovaginal prolactin is not a sensitive test compared with the beta-hCG test. PMID- 11728483 TI - Misoprostol versus oxytocin in the third stage of labor. AB - OBJECTIVES: A double blind randomized controlled trial was performed at the tertiary hospital in Harare, Zimbabwe to compare oral misoprostol with intramuscular oxytocin in the management of third stage of labor. METHODS: A total of 499 women were randomized to receive either 400 microg misoprostol orally or 10 IU oxytocin intramuscularly. The incidences of postpartum hemorrhage and side effects were examined. RESULTS: The demographic and labor characteristics were comparable. Postpartum hemorrhage occurred in 15.2% of women given misoprostol and in 13.3% of those given oxytocin (P=0.534). Measured blood loss of more than 1000 ml occurred in 3.7% of the misoprostol group compared with 2% in the oxytocin group (P=0.237). There was no significant difference in the need for additional oxytocic drugs or blood transfusion in women given misoprostol (P values 0.137 and 0.600, respectively). Significant side effects of misoprostol were shivering [RR=1.32 (95% CI 1.11-1.58); P=0.002) and a rise in temperature [RR=2.02 (95% CI 1.75-2.33); P<0.001]. CONCLUSIONS: Oral misoprostol is as effective as intramuscular oxytocin in the prevention of PPH. Shivering and transient pyrexia were specific side effects of misoprostol. Misoprostol has potential in reducing the high incidence of PPH in developing countries. PMID- 11728484 TI - Elevated C-reactive protein and pro-inflammatory cytokines in Andean women with pre-eclampsia. AB - OBJECTIVE: To investigate the concentration of markers of inflammation in non pregnant women, women with normal pregnancy and women with pre-eclampsia. METHODS: Pregnant women (n=26), women with pre-eclampsia (n=25) and non-pregnant normotensive women (n=21) were included in the study. C-reactive protein was measured by latex-enhanced immunoturbidimetric assay, serum tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) by high sensitivity ELISA. Kruskal Wallis non-parametric analysis of variance followed by the Mann-Whitney U-test were used for statistical analyses. RESULTS: Higher values (mean+/-S.E.M.) of C reactive protein were found in pre-eclampsia (4.11+/-0.37 mg/dl) compared with normal pregnant women (2.49+/-0.26 mg/dl) and non-pregnant controls (1.33+/-0.15 mg/dl). TNF-alpha was significantly higher in women with pre-eclampsia (15.74+/ 5.09 pg/ml), in relation to the control group (2.76+/-0.41 pg/ml) and women with normal pregnancy (8.31+/-1.55 pg/ml). IL-6 levels were significantly higher in pre-eclamptic women (12.91+/-1.29 pg/ml) compared with normal pregnant (5.07+/ 0.423 pg/ml) and control women (1.25+/-0.13 pg/ml). CONCLUSIONS: The results of this cross-sectional study in a high-risk Andean population show that both C reactive protein and pro-inflammatory cytokines are present in higher concentrations in women with pre-eclampsia. The study was undertaken in women with established pre-eclampsia and it is not possible to determine whether the increased concentrations of C-reactive protein and pro-inflammatory cytokines were a cause or consequence of the disease. PMID- 11728485 TI - Colposcopic evaluation after a repeat atypical squamous cells of undetermined significance (ASCUS) smear. AB - OBJECTIVES: Management of patients with atypical squamous cells of undetermined significance (ASCUS) remains controversial. We chose to repeat the Pap smear after four months. If ASCUS persisted in this second test, the patient was advised to undergo colposcopy. Our objective is to determine the clinical significance and the prediction of neoplasia among these patients through a colposcopic examination. METHODS: Of 29,827 patients who had a Pap smear, ASCUS were detected in 1387 (5%) and persisted in the repeat smear of 225 (16%). Colposcopy and an additional Pap smear were performed on 186 patients. RESULTS: Out of 186 colposcopic evaluations, 91 (49%) were normal and the patients had a negative Pap smear. Colposcopy was abnormal in 95/186 patients (51%) (Table 1). Histology of the directed biopsies revealed 38 (21%) low-grade squamous intraepithelial lesions (LSIL) and 17 (9%) high-grade squamous intraepithelial lesions (HSIL). Forty patients (21%) with normal biopsies had ASCUS for the third time in the Pap smear. CONCLUSIONS: Colposcopic evaluation after a repeated Pap smear with ASCUS is an appropriate cost-effective management. Finding 30% of LSIL or HSIL justifies this additional investigation. PMID- 11728487 TI - Bacterial adherence and immune response in recurrent urinary tract infection. AB - OBJECTIVE: To study the significance of bacterial adherence and humoral immune response in the pathogenesis of recurrent urinary tract infection in adult women. METHODS: The study population consisted of 30 women with recurrent urinary tract infection (UTI) and 30 healthy women as controls. Bacterial adherence to vaginal epithelial cells was studied. The concentration of serum and urine antibodies against mixed coliform antigen and clinical isolate was determined by enzyme linked immunosorbant assay (ELISA). The surface hydrophobicity of the organisms was determined. Student's unpaired t-test and Pearson's correlation coefficient test were used in the statistical analysis. RESULTS: There was an increased adherence of bacteria to vaginal epithelial cells of women with recurrent UTI. The SIgA level in urine was low compared with other antibodies. There was good correlation between the level of free urinary IgG antibody and bacterial adherence. CONCLUSIONS: This study showed that increased receptivity of vaginal epithelial cells to pathogens and lower local immunity may play an important role in the pathogenesis of recurrent UTI in females. PMID- 11728486 TI - Endometrial histologic changes in post-menopausal breast cancer patients using tamoxifen. AB - OBJECTIVE: The aim of this study was to evaluate the effect of tamoxifen on the endometrium of post-menopausal women with breast cancer and to examine the relationship between ultrasonography, hysteroscopy and histopathologic changes. METHOD: Included in this longitudinal study were 303 post-menopausal women taking 20 mg daily of tamoxifen. Hysteroscopy was performed in 83 patients with an endometrial thickness of only >or=5 mm and 34 with vaginal bleeding also. Forty five asymptomatic patients (control group) underwent hysteroscopies. RESULT: The most frequent outcome in patients with endometrial thickness of only >or=5 mm was an atrophic endometrium in an empty cavity (79.5%) whereas simple hyperplasia (35.3%) was found in women with vaginal bleeding. Carcinoma was diagnosed in seven cases (5.9%). In the control group, no endometrial cancer was found. CONCLUSION: This study suggests that patients with a thickness >5 mm should be offered a whole hysteroscopic evaluation, whenever bleeding is reported. PMID- 11728488 TI - Female genital tuberculosis and infertility. AB - OBJECTIVES: This study was performed to evaluate the rate of diagnosed female genital tuberculosis and its presentational symptoms and methods of diagnosis. METHODS: A total of 3088 cases of tuberculosis (TB) who had been registered and treated in the Health Center of Fars Province from 1989 to 1999 were retrospectively studied. From this group, 46 women were diagnosed as having genital TB. The diagnosis in 41 cases was based on the standard pathological criteria of tissue specimens. The other five cases were excluded from this study due to the lack of classical diagnostic criteria. Statistical analysis was performed using the Z-test. RESULTS: The mean age of the patients at the time of diagnosis was 30.4 years. Seven patients presented with abdominal or pelvic pain (17.07%). In this group three cases underwent laparatomy due to abdominal mass and four patients for tubo-ovarian abscess, which led to the diagnosis. Abnormal uterine bleeding was the cause of diagnostic dilatation and curettage in three other patients (7.31%). However, in 31 cases (75.6%) TB was diagnosed during studies performed to evaluate the cause of their infertility, and the most common diagnostic procedure was endometrial curettage (25 cases). Female genital TB accounted for 1.32% of all tuberculous patients in this study. Of these, 75.6% were infertile by definition (Z=12.13 P<0.0001). TB endometritis was detected in 72.03%, tubal involvement in 34.03%, ovarian TB in 12.9% and cervical TB in 2.4% of the patients. CONCLUSIONS: This study confirms the presence of a strong relationship between genital TB and infertility; therefore genital TB would be more frequently diagnosed if this possibility was considered in the evaluation of every infertile patient in areas where tuberculosis is endemic. PMID- 11728489 TI - Hormone replacement therapy to improve left ventricular diastolic functions in healthy postmenopausal women. AB - OBJECTIVES: To investigate the effects of estrogen and estrogen plus progesterone replacement therapy on left ventricular systolic and diastolic function parameters in healthy postmenopausal women. METHODS: Forty-six healthy consecutive postmenopausal women were prospectively enrolled. Hormone replacement therapy (HRT), which was either 0.625 mg/day conjugated equine estrogen (CEE) alone, or with 2.5 mg/day medroxyprogesterone acetate (MPA) was administered depending on the hysterectomy status. Left ventricular systolic and diastolic function parameters were evaluated with echocardiography before and after 6 months of HRT. The paired t-test was used for statistical analysis. RESULTS: Estrogen or estrogen plus progesterone did not significantly alter the left ventricular dimensions and systolic function parameters. However, significant improvements were noted in several diastolic function parameters including late diastolic mitral flow velocity, ratio of early to late mitral flow velocity and isovolumic relaxation time (P=0.003, P=0.001 and P=0.005, respectively, for the CEE group; all P<0.001 for the CEE+MPA group). CONCLUSIONS: Estrogen or estrogen plus progesterone replacement therapy may significantly improve left ventricular diastolic functions in healthy postmenopausal women. PMID- 11728490 TI - Laminaria, induced fetal demise and misoprostol in late abortion. AB - OBJECTIVES: To analyze and determine the safety and effectiveness of induced fetal demise as an adjunctive method in outpatient abortion for patients with advanced pregnancies and to evaluate the independent effect of intrauterine misoprostol administered after amniotomy in late abortion. METHODS: During a 9 year period, 1677 abortions were performed for patients whose pregnancies ranged from 18 through 34 menstrual weeks in an outpatient facility. Of these, 832 were performed by one physician. Techniques for performing all the abortions included induction of fetal demise by intrauterine fetal injection of digoxin and/or hyperosmolar urea, serial multiple laminaria treatment of the cervix, amniotomy, oxytocin induction of labor, and assisted delivery or surgical evacuation of the fetus and placenta. In the last 411 of the 832 patients whose abortions were performed by one physician, misoprostol was placed in the lower uterine segment following amniotomy in order to enhance labor induction, cervical ripening, and fetal expulsion. RESULTS: Of the entire group of 1677 cases, the median gestational age was 22 menstrual weeks. The median procedure time for all cases was 10 min. Measured median blood loss was 125 ml. Blood loss and procedure time increased with length of gestation, but these were not affected by misoprostol. There were three major complications (0.2%) in the overall series. Among patients seen by one physician (N=832), amniotomy-to-procedure time was shorter by 26 min for patients receiving misoprostol, and there was 27% more variability in amniotomy-to-procedure time among patients not receiving misoprostol. Complication rates for patients receiving misoprostol were the same as for those not receiving misoprostol. There were no major complications in the 832 patients seen by one physician, no uterine rupture or perforations, and no cervical lacerations. CONCLUSIONS: Outpatient abortion may be performed safely from 18 through 34 menstrual weeks using combined surgical and medical procedures. Use of intrauterine post-amniotomy misoprostol was associated with reduced amniotomy-to procedure time and reduced variability in the amniotomy-to-procedure time. PMID- 11728491 TI - Recommendations for renovating an operating theater at an emergency obstetric care facility. AB - The importance of emergency obstetric care (EmOC) in reducing maternal mortality has focused attention on both the skills of the clinicians to provide high quality care and on the health facilities in which the care is provided. Essential elements of EmOC include the capacity to perform cesarean sections for which an operating theater is needed. This article focuses on renovation of existing operating theaters to meet the necessary standards. While building, adding to, or renovating operating theaters can be expensive, this article emphasizes appropriate materials that are likely to be locally available and relatively inexpensive. The importance of proper maintenance is discussed. PMID- 11728493 TI - Efficient and effective emergency obstetric care in a rural Indian community where most deliveries are at home. AB - OBJECTIVES: Most life threatening obstetric complications require hospital treatment to avert maternal mortality. Some assume that in developing countries hospital service for the poor must be in government hospitals and that a large proportion of deliveries needs to be in these hospitals to provide timely access to emergency care. This presents a major problem in countries like India, where almost all rural deliveries are at home and accessible government hospitals generally do not provide surgical treatment for obstetric emergencies. The study's objective was to determine obstetric outcomes, patterns and costs of obstetric care in a part of rural Maharashtra, India, where obstetric outcomes appear relatively good even though most deliveries are at home and government hospitals do not provide emergency obstetric care (EmOC). METHODS: 2905 pregnancies were identified and followed to term to learn the number and types of complications, where these complications were treated, how many women received EmOC and how these services affected outcome. RESULTS: Eighty-five percent of 2861 deliveries after 24 weeks were at home. A total of 14.4% of deliveries after 24 weeks had identified complications. Of these complicated deliveries, 78.9% were in a hospital. Forty-eight percent of hospital deliveries were in a private hospital, 35% in our project hospital and 18% in a government hospital. Hospitalized patients with obstetric complications constituted 11.4% of all deliveries. The cesarean section rate for all deliveries was 2.0%. Twenty-two of the cesareans were in private hospitals, 32 in our hospital and four in a government hospital. Hospital case fatality (deaths of mothers with identified complications) was 0.3%. Overall case fatality was 0.5%. However, there were only two maternal deaths from obstetric causes (70 per 100,000 live births), making these rates less than robust. The perinatal mortality rate was 36 per thousand live and still births. These outcome and process indicators are better than those reported in most of India, but both maternal deaths could have been prevented by early referral to hospital and 64% of perinatal deaths were to infants delivered at home. CONCLUSIONS: A network of private clinics with a voluntary, low cost hospital is providing effective EmOC in a remote rural area at very low per capita cost in the absence of easily accessible government service and with only 15% of deliveries in hospitals. Charges are low but low per capita cost is primarily due to intelligent self-selection of patients who need hospital care. Even though overall cost is low, cost is still an important barrier for many poor families. Improving the purchasing power of poor families through insurance or subsidy could be a more effective way to improve EmOC than trying to improve inadequate government facilities. PMID- 11728495 TI - First trimester Down Syndrome screening by nuchal translucency in a Thai population. PMID- 11728496 TI - Labor outcomes of short multiparous women with a previous successful vaginal delivery. PMID- 11728497 TI - The effect of air pollution on the outcome of pregnancies. PMID- 11728498 TI - Non-closure of visceral peritoneum at abdominal hysterectomy. AB - Our purpose was to determine whether non-closure of the visceral peritoneum alters post-operative outcome at abdominal hysterectomy. A prospective, randomized trial was performed. Of the 100 evaluable subjects, 50 had the visceral peritoneum left open whereas 50 were closed. Patients were observed for evidence of morbidity after the operation. Analysis of data was performed with use of the unpaired t-test for continuous variables. The visceral peritonization process at abdominal hysterectomy does not provide immediate post-operative benefits while unnecessarily lengthening surgical time and anesthesia exposure. We suggest that visceral peritonization process can be abolished at abdominal hysterectomy. PMID- 11728499 TI - Decrease in female genital mutilation among Nigerian Ibo girls. PMID- 11728500 TI - Domestic violence among women seeking post-abortion care. PMID- 11728501 TI - FIGO society survey: acceptance and use of new ethical guidelines regarding induced abortion for non-medical reasons. AB - OBJECTIVE: FIGO's Ethical guidelines regarding induced abortion for non-medical reasons offer guidance concerning women's right to safe abortion services and the medical community's attendant responsibilities. Ipas surveyed FIGO constituent societies to determine their agreement with the Guidelines' recommendations and their readiness to use them to improve and expand services. METHOD: Ten months after the Guidelines publication in IJGO, a ten-item questionnaire was mailed to 283 Officers of the 101 FIGO societies, with follow-up prompts to non respondents. RESULTS: Officers of 59 societies responded, divided evenly between those in countries whose laws permit induced abortion on non-medical grounds and those in countries prohibiting it. In 'permitting' countries all responding societies supported the recommendations, and 85% said they should adopt them or had already done so. Two-thirds in 'prohibiting' countries supported the recommendations, but less than half believed their FIGO society, or their government, should adopt them. However, 20% in the 'prohibiting' countries had adopted or formally considered the recommendations and 23% had already brought them to the attention of their governments. CONCLUSION: The FIGO constituent societies showed overall strong support for the recommendations, but efforts need to be made to encourage those in 'prohibiting' countries to promote implementation of the recommendations. PMID- 11728502 TI - The role of age at asthma diagnosis in the development of triggers of asthma episodes. AB - This study examined the relationship of age at asthma diagnosis to the subsequent impacts of 12 common asthma triggers, which we classified as either mainly physically based or strongly psychosocially mediated. The physically based triggers were air pollution, cigarette smoke, high humidity, high/low environmental temperature, allergy problems, respiratory infection, physical activity, and nighttime hours; the psychosocially mediated triggers were stress or worry, anger, excitement, and laughter. Data were collected with questionnaires from families with asthmatic children (n=115), 2 to 20 years of age, as part of a larger study of biological and psychosocial factors in asthma and other illnesses. Using parents' reports, we classified the children as early diagnosed (before age 2) or later-diagnosed (at or after 2) for asthma and compared these groups, separated by gender, in 2 x 2 multivariate analyses. The impacts of all four psychosocially mediated triggers on asthma attacks were significantly greater for the later-diagnosed children than the early-diagnosed children. No age of diagnosis differences were found for any of the physically based triggers, and no gender or interaction effects were found for either type of trigger. PMID- 11728503 TI - Gender differences in psychological distress in adults with asthma. AB - OBJECTIVES: We sought to examine whether there were gender differences in the relationship of depressive, anxiety-related, and somatic symptomatology to the presence, severity, and duration of asthma. METHODS: A total of 54 adult asthma patients (24 women, 30 men) and 31 healthy subjects (19 women, 12 men) were studied. Within each gender group, patients' psychological distress levels were compared as a function of severity (mild vs. moderate) and duration of disease (<5 vs. 5+ years), to each other and with healthy subjects. Data were collected in Ukraine utilizing the Minnesota Multiphasic Personality Inventory (MMPI), the best-validated psychological assessment instrument in Eastern European populations. RESULTS: Relative to healthy women, women with both mild and moderate asthma showed elevated distress in multiple domains reflecting somatic and psychological complaints. In contrast, only men with relatively more severe disease of longer duration showed elevated symptomatology relative to healthy men, with depressive symptoms predominating. CONCLUSION: To the extent that distress in response to asthma takes a more diffuse form and may be expressed at milder and earlier stages of the disease in women than men, the findings suggest the need to tailor asthma education and behavioral interventions to the unique psychological needs of women and men in order to be maximally effective. PMID- 11728504 TI - The structure of symptom report in asthma: a reevaluation. AB - OBJECTIVES: We studied the structure of symptom report in a sample of British asthma patients using the Asthma Symptom Checklist (ASC). METHODS: The ASC was administered to 193 patients, together with a questionnaire on demographics and asthma-related information and the Perceived Control of Asthma Questionnaire. RESULTS: Principal Component Analysis yielded evidence for a six-dimensional structure of the ASC, with positively correlated subscales for panic-fear, irritation, obstruction-dyspnea, obstruction-congestion, fatigue, and hyperventilation symptoms. Individual subscales showed good to excellent item characteristics and internal consistencies in individual subscales. Panic-fear and obstruction-congestion were related to self-reported duration of oral corticosteroid intake. Other ASC subscales were positively related to aspects of health care utilization, limitations in daily activity, and lower perceived control of asthma mainly in women. The dyspnea subscale of the ASC showed only little relationship with either of these variables. CONCLUSION: The ASC can reliably assess the multidimensional structure of asthma symptom report. Its subscales are associated with important aspects of asthma management behavior. PMID- 11728505 TI - Effects of oral contraceptives on daily self-ratings of positive and negative affect. AB - OBJECTIVES: The relationship between affect and duration of oral contraceptive (OC) use was investigated. METHOD: Ninety-six women (17 first-time OC users, 34 long-time users, and 45 never-users) completed the Positive and Negative Affect Schedule (PANAS) and the Menstrual Distress Questionnaire (MDQ) daily for 35 days. This study was the first to examine positive affect variability; and personal family psychiatric history; and to compare early-, late-, and never users of OCs. RESULTS: Triphasic users experienced greater variability in positive affect across the cycle, likely due to the variable hormone levels. Withdrawal of a constant level of hormones (monophasics) during early use was associated with greater variability in positive affect than withdrawal of changing hormonal levels (triphasics). Furthermore, personal and family psychiatric history may mediate an effect of OCs on negative affect variability. CONCLUSIONS: OCs and, therefore, hormones can alter day-to-day affect variability. Four variables are associated with this effect: duration of use, OC type, personal psychiatric history, and family psychiatric history. PMID- 11728506 TI - Does the Edinburgh Postnatal Depression Scale measure anxiety? AB - OBJECTIVE: The existence of a separate anxiety and depression dimension within the Edinburgh Postnatal Depression Scale (EPDS) has been reported previously. However, the concurrent validity of this anxiety subscale was never evaluated. We investigated whether (1) this existence of an anxiety subscale could be confirmed and (2) it more highly correlated with other measures of anxiety than the total EPDS. METHODS: The SCL-90-R, the EPDS, and the State-Trait Anxiety Inventory (STAI) were filled out by 197 pregnant women. A principal component analysis (PCA) was used for confirmation of the subscales and correlations were computed between the (subscales of the) EPDS and the other measures of anxiety. RESULTS: The existence of an anxiety scale within the EPDS was confirmed. However, this subscale did not yield higher correlations with other measures of anxiety than did the total EPDS. CONCLUSION: Investigators using the EPDS to screen for depression should realise that the instrument does not exclusively measure depression. It seems that both anxiety symptoms and depressive symptoms are more accurately measured when using the total 10-item EPDS than when using the subscales. PMID- 11728507 TI - Insulin-treated diabetes patients with fear of self-injecting or fear of self testing: psychological comorbidity and general well-being. AB - OBJECTIVE: To examine psychological functioning and self-management behaviours of Dutch adult patients with insulin-requiring diabetes mellitus suffering from extreme fear of self-injecting (FSI) and/or fear of self-testing (FST). METHODS: A cross-sectional survey was performed in a sample of insulin-treated diabetes patients (n=1275; 51.1% male; age 49.7+/-15.8 years; 58.0% Type 1 diabetes), assessing FSI and FST. Patients completed the questionnaires concerning trait/state anxiety, depression, fear of hypoglycemia, diabetes-related distress, diabetes self-care activities, and general well-being. Comparisons were made on these measures between patients with extremely high scores on FSI and/or FST (> or = 95th percentile) and the other patients. Patients with extreme scores on FSI and/or FST were invited to take part in a second survey to assess the prevalence of major depression, common fears/phobias, and psychoneuroticism. RESULTS: People with extreme FSI/ FST scores, as compared to the other patients, reported higher levels of trait/state anxiety and depression. This group also reported more fear of hypoglycaemia and diabetes-related distress, had lower levels of general well being, and reported less frequent self-monitoring of blood glucose. The second survey showed 11.1% of patients with extreme FSI/FST reporting scores indicating major depression. Prevalence of scores greater than or equal to the high scores on phobias (38.0-63.3%) and psychoneuroticism (27.8%) were consistently higher than norm group prevalences. DISCUSSION: Extreme levels of FSI and/or FST are associated with high diabetes-related distress, poor general well-being, and psychological comorbidity, as well as poorer adherence to the diabetes treatment regimen. It is concluded that patients with extreme FSI/FST are often burdened with more than this specific phobia. PMID- 11728508 TI - Comorbidity of diabetes mellitus and eating disorders: a follow-up study. AB - OBJECTIVE: There is increasing evidence that the coexistence of diabetes and eating disorders (ED) leads to poor glycemic control and an increased risk of long-term complications. METHODS: In a questionnaire- and interview-based study, a sample of 36 out of originally 38 (94.7%) diabetic patients with an ED (type-1: n=13, type-2: n=23) was assessed after a period of about 2 years in order to determine the course of EDs, body mass index (BMI), glycemic control, and psychiatric symptomatology. RESULTS: Five patients (13.9%) of the total sample showed full remission for at least 12 consecutive weeks. Twenty-two patients (61.6%) showed no change in the diagnosis of the ED. Four patients (11.1%) shifted from subclinical to clinical EDs and five patients (13.9%) vice versa. Of the eight patients who went on to psychotherapy, only one patient (12.5%) showed full remission. Emotional distress of type-2 diabetics was considerably higher compared to type-1 diabetics, which was rather low at baseline. Except interpersonal distrust as one ED-related variable, no significant change of any psychological variable could be observed in the type-1 diabetic sample during follow-up. Of the 13 type-1 diabetic patients with an ED, five patients deliberately omitted insulin in order to lose weight. These patients showed a more serious psychopathology with regard to each measured psychological variable, a higher BMI, and worse metabolic control compared to those without insulin omission. Type-2 diabetics showed a significant increase in drive for thinness and body dissatisfaction. No considerable change could be observed with regard to BMI, glycemic control, and depressive and global psychiatric symptomatology in either diabetic subsample during follow-up. CONCLUSION: EDs tended to persist over time with a considerable shift within the different types of EDs. Insulin purging in type-1 diabetics was associated with enhanced psychopathology, higher BMI, and worse metabolic control. Both mean body mass and ED-related symptoms such as "drive for thinness" and "body dissatisfaction" increased in the average obese type-2 diabetic sample, illustrating the vicious circle of low self-esteem, enhanced restraint eating, and binge eating in weight control measures. PMID- 11728509 TI - Information processing in chronic fatigue syndrome: a preliminary investigation of suggestibility. AB - This study examines the effects of certain types of information processing on the subjective experience of cognitive deficits in persons with chronic fatigue syndrome (CFS). Two groups of participants, persons with CFS and a group of healthy controls, were administered a symptom inventory and measures of intellectual functioning, memory, automatic processing, and suggestibility. The groups differed significantly on number and severity of reported symptoms and on measures of global suggestibility and automatic processing, but not on measures of intellectual functioning and memory. Suggestibility was related to number and severity of reported symptoms, as well as the inability to inhibit the automatic processing of information. Implications of these findings are discussed, as well as directions for future research and treatment of symptoms associated with CFS. PMID- 11728510 TI - Personality and peritraumatic dissociation in the prediction of PTSD in victims of road traffic accidents. AB - OBJECTIVES: To investigate the contribution of personality and peritraumatic dissociation in the development of posttraumatic stress disorder (PTSD). METHOD: Victims of road traffic accidents (RTA) were assessed within 2-4 weeks (Time 1) of the accident and again between 4 and 6 months (Time 2). The Eysenck Personality Questionnaire (EPQ) and Peritraumatic Dissociation Experience Questionnaire (PDEQ) were administered at Time 1 and posttraumatic stress symptoms were assessed at Time 2. RESULTS: 265 subjects were assessed at both time points, although neuroticism, psychoticism, and peritraumatic dissociation were significantly correlated with posttraumatic symptoms, only the personality dimensions were independent and significant predictors of subsequent PTSD in a logistic regression. CONCLUSIONS: Consistent with the literature personality measures, especially neuroticism, is associated with the development of PTSD. However, peritraumatic dissociation was not found to be an independent predictor of PTSD. PMID- 11728511 TI - Chronic disease in elderly couples: are women more responsive to their spouses' health condition than men? AB - OBJECTIVE: The aim of the present study is to increase knowledge regarding associations between couples' health condition and psychological distress in both spouses considering gender as well as patient/spouse status. METHOD: We examined a community-based sample of 995 elderly couples in which either both spouses were healthy, one of them suffered from chronic disease or both spouses were ill. Both spouses filled out the Hospital Anxiety and Depression Scale. RESULTS: In line with our hypotheses, the results showed an association between women's psychological distress and their own as well as their spouse's health condition, whereas men's psychological distress was associated only with their own health condition. CONCLUSION: The findings demonstrate the need for awareness of gender and patient/spouse differences in psychological distress among elderly couples confronted with chronic disease. PMID- 11728512 TI - Work-related stress and depressive disorders. AB - The 1980s and 1990s has seen a considerable change in the workforce structure in industrialised economies. Employees are commonly faced with greater demands and less job security, both of which are likely to be stressful, thus psychological disorders especially depression may increasingly be caused by work-related stressors. An issue of this journal in 1997 (Vol. 43, No. 1) was indeed devoted to stress in the workplace and since then, these workplace changes have progressed and a review seems timely. Because interpreting results of cross sectional studies is limited by a potential reciprocal relation between work stressors and depression (since "effort after meaning" can influence how "distressed" individuals report stressors at work), this review largely focuses on prospective or predictive studies to minimise this bias. Not surprisingly, the findings from occupational stress research is consistent with the more general life event stress literature showing that specific acute work-related stressful experiences contribute to "depression" and, more importantly perhaps, that enduring "structural" occupational factors, which may differ according to occupation, can also contribute to psychological disorders. There are significant implications for employees, their families, employers and indeed the wider community. PMID- 11728516 TI - Progesterone-action in the decidual mesometrium of pregnancy. AB - The mesometrial decidua is absolutely dependent on progesterone action for its maintenance and growth. Hormone action is mediated by intranuclear progesterone receptors (PR) that regulate target cell gene transcription. In early pregnancy of the rat gene expression is particularly enhanced for regulators of cell cycle progression, growth factors and their cognate receptors; cell cycle arrest proteins are suppressed. Cell survival proteins such as Bcl2 are also up regulated. These events are associated with abundant expression of PR-A and PR-B isoforms and STAT (signal transducers and activators of transcription) family members. Proliferation of decidual cells no longer occurs after mid-pregnancy despite high levels of circulating progesterone and the decidua begins a slow process of regression, which continues to term. Regression is characterized by an increase in abundance of proteins that promote apoptosis such as p27, Bax and Caspase-3. These late pregnancy changes are associated with a relative increase in PR-C, a third form of the PR molecule, that binds progesterone but probably has limited transcriptional activity. Protein kinase C, which is suppressed by progesterone in early pregnancy, may be a key mediator of these processes. PMID- 11728517 TI - Molecular perspectives on selective estrogen receptor modulators (SERMs): progress in understanding their tissue-specific agonist and antagonist actions. AB - Synthetic estrogen receptor ligands such as tamoxifen and raloxifene produce biologic responses which can be either estrogenic or anti-estrogenic, depending upon the tissue in which their action is examined. To reflect the fact that they are not 'pure' antagonists, such ligands have been more accurately termed selective estrogen receptor modulators (SERMs). Recent progress in our understanding of the molecular biology of estrogen receptor (ER) action has provided a great deal of evidence which promises to increase our understanding of the mechanism through which SERMs elicit their tissue-specific effects. The identification of numerous coactivators and corepressors which modulate receptor function and the realization of two subtypes of ER attest to the potential complexity through which SERMs produce diverse tissue-specific responses. Evidence from co-crystal structures of ER ligand-binding domains complexed with SERMs provides additional information as to how this class of ligands can elicit diverse biologic responses. SERMs also influence the stability of the ER protein, and recent information on the determinants of receptor stability and the role of proteasome-mediated protein degradation in ER-driven transcription also promises to give a fuller understanding of SERM biology. These aspects of the molecular biology of estrogen receptor action may help clarify the mechanism(s) of SERM biologic action and will be addressed in further detail in this review. PMID- 11728518 TI - The effect of isoflavone extract ingestion, as Trinovin, on plasma steroids in normal men. AB - Plasma testosterone, dihydrotestosterone, androstenedione, dehydroepiandrosterone sulfate, androsterone and epiandrosterone sulfates, cortisol and sex hormone binding globulin were measured in six adult men before and during daily isoflavone extract ingestion (40 mg) in the form of Trinovin tablets. Although modest plasma genistein levels were achieved following three weeks of Trinovin ingestion (106-356 nmol/l) there were no significant changes in most of the analytes tested. However plasma levels of dihydrotestosterone showed an increase that reached significance when combined basal levels were compared to levels following Trinovin treatment. The results suggest that the daily ingestion of isoflavones in the form of Trinovin (1 tablet/day), over a short term, does not alter most plasma steroid levels. We therefore question the value of Trinovin, at the recommended dosage, as offering protective effects against prostate disease by mechanisms involving either significant modulation of plasma steroid or SHBG levels. In contrast the increase in dihydrotestosterone plasma levels could be seen as possibly detrimental. PMID- 11728519 TI - 1,5-Hydride shift in Wolff-Kishner reduction of (20R)-3beta,20, 26-trihydroxy-27 norcholest-5-en-22-one: synthetic, quantum chemical, and NMR studies. AB - Heating (20R)-3beta,20,26-trihydroxy-27-norcholest-5-en-22-one (1) with hydrazine and KOH at 160 degrees C completely converted the steroid to a diastereoisomeric mixture of the new (20R,22RS)-27-norcholest-5-ene-3beta,20,22-triols (2). Exclusive formation of 2 suggests that the expected Wolff-Kishner reduction to a methylene group at the C-22 ketone in 1 was diverted to the C-26 position by a 1,5-hydride shift. All attempts under acid conditions failed to produce a C-22 phenyl hydrazone from 1. However, reaction of 1 was reacted with phenylhydrazine in hot KOH, gave the C-26 phenylhydrazone 4 as the sole product. Evidently, under alkaline conditions, first a hydride ion undergoes an intramolecular transfer from the C-26 CH(2)OH group to the C-22 ketone in 1, and then the phenylhydrazine traps the newly formed aldehyde. To examine this hypothesis, we constructed computer-simulated transition state models from quantum chemical calculations and then compared data from these models with NMR measurements of the reaction mixtures containing 2. The NMR data showed that the C-22 diastereoisomers of 2 are formed in a nearly 1:1 ratio exactly as predicted from the energy-optimized transition states, which were calculated for intramolecular 1,5-hydride shifts that produced each of the two C-22 diastereoisomers. Accordingly, these results support the hypothesis that an intramolecular 1,5-hydride shift mechanism promotes complete conversion of 1 to 2 under classical Wolff-Kishner reduction conditions. PMID- 11728520 TI - Plasma and urinary markers of oral testosterone undecanoate misuse. AB - Orally administered testosterone undecanoate (TU), an anabolic, androgenic steroid, can potentially be abused by athletes. Indirect evidence for detecting oral TU intake could be deduced from the changes in steroid profile post administration. Direct evidence could be obtained by detection of unchanged TU in plasma. To this end, both urinary and plasma steroid profiles of six healthy male subjects given a single oral dose of 120 mg of TU were studied by gas chromatography/mass spectrometry (GC/MS) and gas chromatography/tandem mass spectrometry (GC/MS/MS). The increased concentration of glucuronidated testosterone in plasma appears to be the most characteristic sign of oral TU intake. The testosterone glucuronide (TG)/nonconjugated testosterone (T) ratio, TG/17-hydroxyprogesterone (17OHP) ratio, and TG/luteinizing hormone (LH) ratio were observed to be significantly elevated above their basal levels for 10 h, 10 h, and 6 h, respectively. Urinary ratios of TG/epitestosterone glucuronide (EG) were found to be higher than the cut-off value of 6 for the period 4 approximately 8 h post-administration, but only in three subjects. One subject failed to respond with respect to all of the above-mentioned indirect markers, as TG was not significantly increased in either plasma or urine. Unchanged TU was directly detected in plasma of all six subjects from 1 approximately 1.5 h to 4 approximately 6 h after oral TU intake by GC/MS/MS, providing unequivocal proof of exogenous testosterone intake. Distinct and complementary markers for detecting oral TU intake could be obtained from plasma and urine, respectively. PMID- 11728521 TI - 7alpha-OH epimerisation of bile acids via oxido-reduction with Xanthomonas maltophilia. AB - The microbial 7alpha-OH epimerisation of cholic, chenodeoxycholic, and 12 ketochenodeoxycholic acids (7alpha-OH bile acids) with Xanthomonas maltophilia CBS 827.97 to corresponding 7beta-OH derivatives with scarcity of oxygen is described. With normal pressure of oxygen the 7-OH oxidation products are obtained. No biotransformations are achieved in anaerobic conditions. The microbial 7alpha-OH epimerisation is achieved by oxidation of 7-OH function and subsequent reduction. Partial purification, in fact, of the enzymatic fraction revealed the presence of two hydroxysteroid dehydrogenases (HSDH) alpha- and beta stereospecific together with a glycocholate hydrolase. On the basis of these results a further application is the microbial reduction of 6alpha-fluoro and 6beta-fluoro-3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid methyl esters to the corresponding 7alpha-OH and 7beta-OH derivatives. PMID- 11728522 TI - Analogues of androgen hormones with inverted configuration at carbons 5, 9, and 10. AB - On catalytic hydrogenation of Delta(9)-steroids (e.g. 3beta-hydroxy-5-methyl-19 nor-5beta-androst-9-en-17-one), four isomers were formed: 9alpha,10alpha-, 9alpha,10beta-, 9beta,10alpha- and 9beta,10beta-adducts. The product distribution was affected by the nature of the C-3 substituent. A chair conformation of A, B, and C rings was found in all of the products with the exception of the 9alpha,10alpha-adduct whose B ring adopts a twist boat conformation. The products were utilized for the synthesis of dihydrotestosterone analogues. PMID- 11728523 TI - Coupling of biologically active steroids to conjugating arms through ether linkages for use in immunochemistry. AB - Conjugation of haptens through ether linkages avoids leakage problems in immunoassays, but this procedure is not easily applied to most steroids that bear low reacting hydroxyls. A new technique allowing the ether coupling of biologically active steroids with conjugating arms in mild conditions compatible with thermosensitive protecting groups is presented. In the first step, the solvent (an aromatic hydrocarbon) was dehydrated by azeotropic distillation in a soxhlet apparatus using a cartridge filled with 0.3 nm and 0.4 nm molecular sieves. In this protected medium, a thallium steroid alkoxide was completely formed by reaction of the steroid with thallium ethoxide and by the continuous elimination of ethanol. The halogenated chain was then introduced into the same medium and reacted in the absence of moisture to give the ether. 17beta-Hydroxy and 11alpha-hydroxy derivatives were involved in this reaction. The coupling was effective for all of the compounds tested after 2-36 h of reaction time and at temperatures between 80 and 140 degrees C. The conjugates were at least 95% pure, and yields ranged from 15 to 95%. PMID- 11728524 TI - Patients on peritoneal dialysis but not on hemodialysis have elevated concentration and activity of thrombin-activatable fibrinolysis inhibitor. AB - Cardiovascular disease (CVD) is a leading cause of death in patients on dialysis. Increased concentration of fibrinogen, dyslipidemia and impaired fibrinolysis are regarded as important risk factors for CVD. Thrombin-activatable fibrinolysis inhibitor (TAFI) is a recently discovered inhibitor of the fibrinolytic system. The aim of this study was to investigate whether peritoneal dialysis (PD) and hemodialysis (HD) patients differ with regard to TAFI concentration and/or its activity. We also measured albumin, cholesterol, triglycerides and fibrinogen. The study was performed on 35 chronically dialyzed patients (14 on PD and 21 on HD) and 18 healthy volunteers. TAFI antigen and its activity were measured with commercially available kits. Albumin, cholesterol, triglycerides and fibrinogen were measured using standard laboratory methods. Only PD patients had significantly elevated level of TAFI antigen and its activity compared to control subjects. Differences in TAFI concentration and its activity between PD and HD were at the level of statistical significance (P=.09 and P=.07, respectively). PD patients had significantly higher concentration of cholesterol and triglycerides than HD group. Fibrinogen was elevated significantly in PD patients compared to HD and controls. There was no difference in albumin concentration between PD and HD. Significant positive correlations were found between fibrinogen or triglycerides and TAFI activity only in PD patients. We conclude that the above phenomenon may predispose PD patients to suppression of fibrinolysis. PMID- 11728525 TI - Synergistic outside-in regulation of platelet activation by GPIIb/IIIa ligand induced conformation and oligomerization. AB - Full platelet activation with serotonin secretion and thromboxane A(2) (TxA(2)) formation induced by a low dose of thrombin receptor agonist peptide (TRAP) or high dose ADP requires platelet aggregation. This requirement can be replaced by pretreatment of platelets with a combination of reagents including: GPIIb/IIIa inhibitors yielding ligand-induced binding sites (LIBS), either arginine-glycine aspartate-serine (RGDS) peptide or Ro 43-5054, cytochalasin to disrupt actin filaments and crosslinking by a GPIIb/IIIa mAb (pl-62). Crosslinking is required since Fab fragments of pl-62 do not support activation. Engagement of the Fc receptor by the mAb Fc domain is not required for pl-62 augmentation, since it is not blocked by the anti-Fc receptor mAb, IV-3. Another GPIIb/IIIa inhibitor, Ro 44-9883, not yielding LIBS epitopes, serves as a negative control and shows a requirement for LIBS in addition to crosslinking. Focal adhesion kinase tyrosine phosphorylation induced by TRAP is blocked by these GPIIb/IIIa antagonists, but restored by pl-62 crosslinking independent of LIBS induction. Tyrosine phosphorylation of a peptide comigrating with p38 MAP kinase is also inhibited by these antagonists and restored by pl-62 crosslinking. However, p38 MAP kinase activation by low dose TRAP is not affected by these aggregation inhibitors. Tyrosine phosphorylation of a 34-kDa phosphoprotein in the absence of aggregation or TxA(2) formation was uniquely augmented by Ro 43-5054 but not Ro 44-9883 under the above activation conditions. PMID- 11728526 TI - Antithrombotic effects of YS-49 and YS-51--1-naphthylmethyl analogs of higenamine. AB - The antiplatelet and antithrombotic effects of YS-49 and YS-51--l-naphthylmethyl analogs of higenamine, which is a benzyl-tetrahydroisoquinoline alkaloid isolated from Aconitum japonicum (Ranunculaceae)--were investigated. YS-49 and YS-51 showed inhibitory activities to both human and rat platelet aggregation induced by ADP, collagen and epinephrine. They were more inhibitory to epinephrine induced aggregation (IC(50); 3.4 and 1.7 microM of YS-49, and 6.0 and 6.3 microM of YS-51 to human and rat platelets, respectively) than ADP- or collagen-induced aggregation. The antithrombotic effects of YS-49 and YS-51 were also observed in both mouse acute thrombosis model and rat arterio-venous shunt (AV shunt) model. The oral administration of YS-49 and YS-51 (50 or 100 mg/kg) increased the recovery rates from the acute thrombotic challenge in mice and lowered the weight of thrombus formed inside the AV shunt tube in rats. PMID- 11728527 TI - A new factor X defect (factor X Padua 3): a compound heterozygous between true deficiency (Gly(380)-->Arg) and an abnormality (Ser(334)-->Pro). AB - We report a novel mutation in Factor X (FX) gene which results in a phenotype without any bleeding tendency. The proband has been found to be a compound heterozygote between a novel FX true deficiency (Gly(380)-->Arg) and a previously reported dysfunctional mutation Ser(334)-->Pro (FX Marsiglia). Prothrombin time (PT) and partial thromboplastin time (PTT) were moderately prolonged and were fully corrected by the addition of normal serum. Her FX activity level varied between 8% and 19% of normal according to the method used whereas the FX antigen level was 40% of the normal control value. All the exons and intron/exon junctions of the FX gene were studied using a combined approach of polymerase chain reaction and conformation sensitive gel electrophoresis. A transversion G to A in exon 8 resulting in the replacement of Gly380 by Arg was found in the proband, in the father and in a proband's brother, whereas heterozygous FX Marsiglia was present in the proband's mother and her sister. Gly380 is strictly linked to Ser379, a component of the catalytic triad. The substitution of Gly for Arg causes the introduction of a large charged amino acid which could affect the catalytic function of FX leading to secretion problem, accounting for the cross reactive material (CRM) negative phenotype. PMID- 11728529 TI - The evaluation of clotting times in the laboratory detection of lupus anticoagulants. AB - Although there is international consensus regarding the general principles of testing for lupus anticoagulants (LAs), no agreement exists as far as the analysis of the clotting time results is concerned. Twenty-nine laboratories participating in the Fifth International Survey of Lupus Anticoagulants (ISLA-5) reported the activated partial thromboplastin time (APTT)-based clotting times obtained on seven defined test samples and a normal plasma (NP) using the same two reagents with low and high phospholipid (PL) concentrations, respectively. These clotting times were used to analyse how various methods of calculating the results may influence the apparent sensitivity of LA tests. We found that the use of a separate screening test may lead to the exclusion of samples where the presence of LA would have been detected by a combined screening and confirmatory method. For instance, the dilute APTT (dAPTT) gave a sensitivity of 53.5% (screening test), while the calculation of a ratio between the clotting times obtained with two different PL concentrations gave a sensitivity of 68.1% (confirmatory test). The normalisation of results by dividing with the corresponding results of NP increased the apparent sensitivity. The screening test ratio between dAPTT results of test samples and NP gave a sensitivity of 84.7%. The normalised ratio between the clotting times obtained with the two reagents (lupus ratio, LR) gave a sensitivity of 95.1%. We conclude that when testing for LA, all samples should be tested with both low (screening procedure) and high (confirmatory procedure) PL concentrations. These two clotting times should be evaluated in relation to each other and to the corresponding results obtained with a reference plasma (normalisation). PMID- 11728528 TI - Arginine inhibits hemostasis activation. AB - BACKGROUND: The diagnosis and the therapy of in vivo hemostasis activation is of great clinical importance. Artefactual changes of the hemostasis (i.e., coagulation or fibrinolysis) in vitro have to be prevented. Usual in vitro anticoagulation by sodium citrate does not fully inhibit coagulation--or fibrinolysis--activation. Therefore, there is need for a simple physiologic inhibitor of hemostasis activation both in diagnosis and therapy of hemostasis activation. METHODS: Whole blood clotting time (WBCT), prothrombin time (PT), activated partial thromboplastin time (APTT), in vitro bleeding test closure time (IVBT-CT), and whole blood aggregometry (WBA) were determined in normal human blood or plasma, supplemented with increasing concentrations of L-arginine or guanidine. RESULTS: Arginine in concentrations of 5-100 mM inhibited the WBCT, PT, APTT, IVBT-CT, and WBA. Arginine (50 mM) resulted in a two-fold prolongation of WBCT, PT, or IVBT-CT (the anti-epinephrine action is superior to the anti-ADP action), a four-fold prolongation of APTT or a 60% inhibition of WBA. CONCLUSION: L-Arginine (or guanidine) inhibited the activation of hemostasis. Arginine might be used as hemostasis stabilizer both in the diagnosis and therapy of hemostasis activation. The usage of arginine as an in vitro hemostasis inhibitor might be indicated in the diagnosis of hemostasis activation, as occurring in pharmacological thrombolysis or disseminated intravascular coagulation (DIC). The storage of blood or blood products might be improved by arginine stabilization. The amino acid (and nitric oxide precursor) L-arginine could be an interesting new pharmacologic agent to inhibit a pathologic hemostasis activation. PMID- 11728530 TI - Induction of hepatic tissue-type plasminogen activator and type 1 plasminogen activator-inhibitor gene expressions and appearance of their translation products in the bile following acute liver injury in rats. AB - BACKGROUND/AIMS: The plasminogen activator (PA)-plasmin system is primarily involved in fibrinolysis, but is also in the patho/physiological events in which breakdown of extracellular matrices is evoked topically. In this present study, we examined the expression of fibrinolytic factors, tissue-type PA (t-PA) and Type 1 PA inhibitor (PAI-1), in acute liver injury. METHODS: Acute liver injury was produced in rats by the intraperitoneal administration of carbon tetrachloride (CCl(4)). Plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were measured to verify the hepatocellular damage. t-PA and PAI-1 gene expressions were measured by Northern blotting, and the cell type(s) expressing these genes was identified by in situ hybridization. t-PA and PAI-1 levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: A single intraperitoneal administration of CCl(4) caused severe acute parenchymatous liver injury. Both t-PA and PAI-1 gene expressions were induced by the acute liver injury, and plasma t-PA and PAI-1 concentrations were also increased. In situ hybridization studies demonstrated that the hepatocytes were the cells expressing t-PA and PAI-1 genes during the acute liver injury. t-PA was also augmented in the bile, whereas PAI-1 was decreased there. CONCLUSIONS: t-PA and PAI-1 gene expressions are induced in the hepatocytes of rats with acute liver injury. These fibrinolytic factors induced by liver injury may play important roles in liver regeneration. PMID- 11728531 TI - Homozygous 20210G/A prothrombin gene mutation associated with bilateral iliac vein thrombosis: a case report. PMID- 11728532 TI - Paradoxical activation of major platelet receptors in the methadone-maintained patients after single pill of aspirin. AB - BACKGROUND: Chronic drug abuse is an established cause of acute coronary events and sudden death. The association between use of narcotics and platelet abnormalities is well described. However, it is still not clear, how aspirin affects expression of major platelet receptors in chronic drug users with coronary artery disease, especially those recovering in the methadone clinic maintenance program. We sought to compare how a single pill of non-enteric coated aspirin (325-mg) affects human platelets in patients with coronary artery disease dependent on methadone use. METHODS: Data from 30 subjects were analyzed, eight of them were the chronic drug addicts, and participated in a methadone recovery program. Platelets were assessed twice at baseline (pre-aspirin), and after 3-24 hours (post-aspirin). The expression of platelet receptors was determined by using the following monoclonal antibodies: CD31 (PECAM-1), CD41 (GPIIb), CD42b (GPIb), CD51/CD61 (vitronectin receptor), CD62p (P-selectin), CD63 (LIMP or LAMP 3), CD107a (LAMP-1), CD151 (PETA-3), and PAC-1 for fibrinogen-platelet binding determination (PharMingen, San Diego, CA). Platelet-leukocyte interactions were assessed by using dual antibodies for a pan-platelet marker (CD151), together with CD14, a monocyte/macrophage marker. RESULTS: In a drug free group, digestion of a single tablet of aspirin resulted in a significantly (p<0.05) diminished expression of PECAM-1, GP IIb, fibrinogen binding with PAC-1 antibody, GP Ib, P selectin, and CD151. In contrast, patients receiving methadone exhibited opposite trends, resultant in a paradoxical activation of major platelet receptors after digestion of aspirin. These differences reached statistical significance (p<0.05) for PECAM-1, GPIIb, and P-selectin expression. CONCLUSION: There are some fundamental differences in the responsiveness to aspirin in chronic methadone users when compared with drug-free patients. Suspecting narcotics abuse may be critical in patients with limited aspirin efficacy, or those who developed an aspirin resistance. Unexpected platelet activation may indeed represent a missing link between use of narcotics and enhanced incidence of vascular death in this high-risk population. PMID- 11728533 TI - Climate change--the new bioterrorism. PMID- 11728534 TI - Time for total shift to first-trimester screening for Down's syndrome. PMID- 11728535 TI - Development of hypertension: implications for primary prevention. PMID- 11728536 TI - Interaction between sperm and zona pellucida in male fertility. PMID- 11728537 TI - A "bureausceptic" view of cancer drug rationing. PMID- 11728538 TI - Mechanism of interferon resistance in hepatitis C. PMID- 11728539 TI - Improving the image of biotechnology the European way. PMID- 11728540 TI - Absence of nasal bone in fetuses with trisomy 21 at 11-14 weeks of gestation: an observational study. AB - BACKGROUND: Prenatal diagnosis of trisomy 21 requires an invasive test in women regarded as being at high risk after screening. At present there are four screening tests, and for a 5% false-positive rate, the sensitivities are about 30% for maternal age alone, 60-70% for maternal age and second-trimester maternal serum biochemical testing, 75% for maternal age and first-trimester fetal nuchal translucency scanning, and 85% for maternal age with fetal nuchal translucency and maternal serum biochemistry at 11-14 weeks. In this study, we examined the possible improvement in screening for trisomy 21 by examining the fetal nasal bone with ultrasound at 11-14 weeks of gestation. METHODS: We did an ultrasound examination of the fetal profile in 701 fetuses at 11-14 weeks' gestation immediately before karyotyping for a possible chromosomal abnormality detected by maternal age and fetal nuchal translucency screening. The presence or absence of a nasal bone was noted. FINDINGS: The fetal profile was successfully examined in all cases. The nasal bone was absent in 43 of 59 (73%) trisomy 21 fetuses and in three of 603 (0.5%) chromosomally normal fetuses. The likelihood ratio for trisomy 21 was 146 (95% CI 50-434) for absent nasal bone and 0.27 (0.18-0.40) for present nasal bone. In screening for trisomy 21, by a combination of maternal age and fetal nuchal translucency, we estimated that inclusion of examination of the fetal profile for the presence or absence of nasal bone could increase the sensitivity to 85% and decrease the false-positive rate to about 1%. INTERPRETATION: In screening for trisomy 21, examination of the fetal nasal bone could result in major reduction in the need for invasive testing and a substantial increase in sensitivity. PMID- 11728541 TI - Oral triptans (serotonin 5-HT(1B/1D) agonists) in acute migraine treatment: a meta-analysis of 53 trials. AB - BACKGROUND: The triptans, selective serotonin 5-HT(1B/1D) agonists, are very effective acute migraine drugs with a well- developed scientific rationale. Seven different triptans will soon be clinically available, making evidence-based selection guidelines necessary. Triptan trials have similar designs, facilitating meta-analysis; this will provide a foundation for using triptans in clinical practice. METHOD: We asked pharmaceutical companies and the principal investigators of company-independent trials for raw patient data of all double blind, randomised, controlled, clinical trials of oral triptans in migraine. We calculated summary estimates across studies for important efficacy and tolerability parameters, and separately summarised direct comparator trials. RESULTS: 53 clinical trials (12 unpublished) involving 24089 patients, met the criteria for inclusion. Mean results for 100 mg sumatriptan were 59% (95% CI 57 60) for 2 h headache response (improvement from moderate or severe to mild or no pain); 29% (27-30) for 2 h pain free (improvement to no pain); 20% (18-21) for sustained pain free (pain free by 2 h and no headache recurrence or use of rescue medication 2-24 h post dose); and 67% (63-70) for consistency (response in at least two of three treated attacks); placebo-subtracted proportions for patients with at least one adverse event (AE) were 13% (8-18), for at least one central nervous system AE 6% (3-9), and for at least one chest AE 1.9% (1.0-2.7). Compared with these data, 10 mg rizatriptan showed better efficacy and consistency, and similar tolerability; 80 mg eletriptan showed better efficacy, similar consistency, but lower tolerability; 12.5 mg almotriptan showed similar efficacy at 2 h but better other results; 2.5 mg naratriptan and 20 mg eletriptan showed lower efficacy and (the first two) better tolerability; 2.5 mg and 5 mg zolmitriptan, 40 mg eletriptan, and 5 mg rizatriptan showed very similar results. The results of the 22 trials that directly compared triptans show the same overall pattern. We received no data on frovatriptan, but publicly available data suggest lower efficacy. INTERPRETATION: At marketed doses, all oral triptans were effective and well tolerated. 10 mg rizatriptan, 80 mg eletriptan, and 12.5 mg almotriptan provide the highest likelihood of consistent success. PMID- 11728542 TI - Priority-setting decisions for new cancer drugs: a qualitative case study. AB - BACKGROUND: Frameworks for legitimate and fair priority setting emphasise the importance of the rationales for priority setting decisions. However, priority setting rationales, in particular for new cancer drugs, are not well described. We describe the rationales used by a committee making funding decisions for new cancer drugs. METHODS: We did a qualitative case study of a priority setting committee (Cancer Care Ontario Policy Advisory Committee for the New Drug Funding Program) by analysing documents, interviewing committee members, and observing committee meetings. FINDINGS: We identified and described decisions and rationales related to 14 drugs in eight disease conditions over 3 years. Our main findings were that: priority setting existed in relation to resource mobilisation; clinical benefit was the primary factor in decisions; in the context of an expanding budget, rationales changed; rationales could change as costs for individual treatments increased; when all options were reasonable, groups funded a range of options and let patients decide; and priority setting rationales involve clusters of factors, not simple trade-offs. INTERPRETATION: Observing priority-setting decisions and their rationales in actual practice reveals lessons not contained in theoretical accounts. PMID- 11728543 TI - Clinical picture: An unwanted tattoo. PMID- 11728544 TI - Assessment of frequency of progression to hypertension in non-hypertensive participants in the Framingham Heart Study: a cohort study. AB - BACKGROUND: Patients with optimum (<120/80 mm Hg), normal (120-129/80-84 mm Hg), and high normal (130-139/85-89 mm Hg) blood pressure (BP) may progress to hypertension (>140/90 mm Hg) over time. We aimed to establish the best frequency of BP screening by assessing the rates and determinants of progression to hypertension. METHODS: We assessed repeated BP measurements in individuals without hypertension (BP<140/90 mm Hg) from the Framingham Study (4200 men, 5645 women; mean age 52 years) who attended clinic examinations during 1978-94. The incidence of hypertension (or use of antihypertensive treatment) and its determinants were studied. FINDINGS: A stepwise increase in hypertension incidence occurred across the three non-hypertensive BP categories; 5.3% (95% CI 4.4-6.3%) of participants with optimum BP, 17.6% (15.2-20.3%) with normal, and 37.3% (33.3-41.5%) with high normal BP aged below age 65 years progressed to hypertension over 4 years. Corresponding 4-year rates of progression for patients 65 years and older were 16.0% (12.0-20.9), 25.5% (20.4-31.4), and 49.5% (42.6 56.4), respectively. Obesity and weight gain also contributed to progression; a 5% weight gain on follow-up was associated with 20-30% increased odds of hypertension. INTERPRETATION: High normal BP and normal BP frequently progress to hypertension over a period of 4 years, especially in older adults. These findings support recommendations for monitoring individuals with high normal BP once a year, and monitoring those with normal BP every 2 years, and they emphasise the importance of weight control as a measure for primary prevention of hypertension. PMID- 11728545 TI - HIV-1 and recurrence, relapse, and reinfection of tuberculosis after cure: a cohort study in South African mineworkers. AB - BACKGROUND: The proportion of recurrent tuberculosis cases attributable to relapse or reinfection and the risk factors associated with these different mechanisms are poorly understood. We followed up a cohort of 326 South African mineworkers, who had successfully completed treatment for pulmonary tuberculosis in 1995, to determine the rate and mechanisms of recurrence. METHODS: Patients were examined 3 and 6 months after cure, and then were monitored by the routine tuberculosis surveillance system until December, 1998. IS6110 DNA fingerprints from initial and subsequent episodes of tuberculosis were compared to determine whether recurrence was due to relapse or reinfection All patients gave consent for HIV-1 testing. FINDINGS: During follow-up (median 25.1 months, IQR 13.2 33.4), 65 patients (20%) had a recurrent episode of tuberculosis, a recurrence rate of 10.3 episodes per 100 person-years at risk (PYAR)-16.0 per 100 pyar in HIV-1-positive patients and 6.4 per 100 pyar in HIV-1-negative patients. Paired DNA fingerprints were available in 39 of 65 recurrences: 25 pairs were identical (relapse) and 14 were different (reinfection). 93% (13/14) of recurrences within the first 6 months were attributable to relapse compared with 48% (12/25) of later recurrences. HIV-1 infection was a risk factor for recurrence (hazard ratio 2.4, 95% CI 1.5-4.0), due to its strong association with disease caused by reinfection (18.7 2.4-143), but not relapse (0.58; 0.24-1.4). Residual cavitation and increasing years of employment at the mine were risk factors for relapse. INTERPRETATION: In a setting with a high risk of tuberculous infection, HIV-1 increases the risk of recurrent tuberculosis because of an increased risk of reinfection. Interventions to prevent recurrent disease, such as lifelong chemoprophylaxis in HIV-1-positive tuberculosis patients, should be further assessed. PMID- 11728546 TI - Saint-Anthony's fire. PMID- 11728547 TI - Eradication of Helicobacter pylori and improvement of hereditary angioneurotic oedema. AB - Helicobacter pylori infection is thought to be a causal factor in various dermatological disorders. We assessed the frequency of H pylori infection in 65 patients with hereditary angioneurotic oedema. We measured the serum concentration of antibodies against H pylori and did the carbon-14-urease breath test in patients with positive H pylori serology. 19 of 65 patients had H pylori infection. All patients with infection, and 11 of 46 without infection, had a history of recurrent episodes of acute abdominal pain. We successfully eradicated H. pylori infection in 18 patients. The frequency of abdominal symptoms was significantly higher in the infected group (p=0.002 after adjustment for age). In nine of 19 patients with dyspepsia, the frequency of oedematous episodes decreased from 100 over 10 months before eradication to 19 during the 10-month follow-up period. Screening for, and eradication of, H pylori infection seems to be justified in patients with hereditary angioneurotic oedema. PMID- 11728548 TI - Analysis of hereditary component of cancer by use of a familial index by site. AB - To quantify hereditary factors in the risk of cancer, we matched 1283047 cancer patients listed in the Swedish Cancer Registry with healthy controls from a national database, and identified the number of individuals who were first-degree relatives of other individuals in the same cohort. Division of the number of relatives in the patient cohort by the number in the control cohort yielded a "familial index". The following cancers had high familial indices: eye 16.5 (95% CI 2.5-666), testis 9.0 (3.2-35), Hodgkin's disease 6.5 (2.3-26.0), and thyroid 6.2 (3.7-12). Overall, however, familial factors made only a minor contribution to susceptibility to cancer. PMID- 11728549 TI - Abnormal microcirculation in brain tumours during surgery. AB - Orthogonal Polarisation Spectral imaging is a new clinical technique that uses polarised light to visualise the microcirculation of organ surfaces. We have used this technique to compare the functional microcirculatory properties of three types of brain tumour with those of the normal cortex. Meningiomas were characterised by chaotic and dilated vessels with almost no erythrocyte movement and a dark background, glioblastoma by few vessels with low flow and a background close to that of the healthy cortex, and metastases by a very dark background, almost no flow, and a chaotic vascular pattern. This technique could provide useful information about microcirculatory morphology and kinetics for diagnostic use during neurosurgery. PMID- 11728550 TI - Intrauterine T-cell activation and increased proinflammatory cytokine concentrations in preterm infants with cerebral lesions. AB - Brain injury is common in very preterm infants, and intrauterine infection is a frequent antecedent of preterm birth. We examined the relation of cerebral damage to intrauterine antigen exposure and inflammation in 50 infants who were born at 23-29 weeks' gestation. Higher concentrations of cytokines (tumour necrosis factor alpha [TNF-alpha], and interleukins [IL], 1beta, 6, and 10) and CD45RO(+) T lymphocytes in umbilical blood predicted cerebral lesions detected by magnetic resonance imaging very soon after delivery. Our results suggest that infants who mount an immune response in utero are at higher risk of cerebral lesions. PMID- 11728551 TI - Human-rights concerns raised about Northern Alliance. PMID- 11728552 TI - Protein chemistry could allow better diabetes control. PMID- 11728553 TI - Haemoglobin C protection against malaria. PMID- 11728554 TI - Molecular decoy may keep bypass grafts open. PMID- 11728555 TI - Early and aggressive treatment saves US anthrax victims. PMID- 11728556 TI - Breathing hard to keep up with HIF-1. PMID- 11728557 TI - Rebuilding the mental-health system. PMID- 11728559 TI - NIH creates registry of stem cells. PMID- 11728560 TI - Patent laws high on the agenda at WTO. PMID- 11728561 TI - New group to look at helping the sick doctors of Europe. PMID- 11728562 TI - Alleged rape of 9-month-old baby shocks South Africa. PMID- 11728563 TI - New technology to combat terrorism set for deployment. PMID- 11728564 TI - Recruitment begins for US smallpox vaccine trial. PMID- 11728565 TI - Insulinotropic meglitinide analogues. AB - The loss of early-phase insulin secretion is an important and early event in the natural history of type 2 diabetes. Because a normal pattern of insulin secretion is essential for the effective control of postprandial metabolism, a rational basis for the development of agents that target early-phase insulin release exists. Conventional oral hypoglycaemic agents do not target, or adequately control, postprandial glycaemia. The emergence of new classes of oral agent with a more specific mode of action provides, for the first time, an opportunity to restore early-phase insulin release. One such drug class is the meglitinide analogues (repaglinide, nateglinide, and mitiglinide). These drugs are ideally suited for combination use with metformin. They could also prove effective in combination with a thiazolidinedione, a drug class that targets insulin resistance. Exogenous insulin is frequently required in the late management of type 2 diabetes. However, one hope for newer combinations of diabetic drugs is that the functional life of the beta cell can be extended, thereby delaying the need for insulin injections. PMID- 11728566 TI - Criminal deterrence as a public health strategy. AB - Deterrence is an established theme in criminal justice, but its role in prevention of assault has been treated with ambivalence and even hostility in medicine. The extent to which offenders can be persuaded, through knowledge of criminal and health risks, not to injure others is emerging from studies of the health effects of firearm and other crime legislation, and from macro-level studies and controlled experiments of police interventions. There is convincing evidence that motorists can be deterred from alcohol-impaired driving, and recognition that specific, targeted, and visible police work and increasing certainty of punishment are effective interventions. By contrast, duration of imprisonment and generic police initiatives such as blanket increases in police numbers seem to have little effect on deterrence, at least in the context of the decline in US homicide rates since 1991, to which demographic and economic factors seem to have contributed little. Together with established and cost effective preschool education and early family support, targeted policing and increasing rates of conviction should be integrated into strategies for injury prevention. PMID- 11728567 TI - Uses of error: Diagnosing tachycardia. PMID- 11728568 TI - Biotechnology and research: promise and problems. AB - Biotechnology has expanded greatly during the last two decades. This expansion is due to worldwide investment in basic biomedical research. The new knowledge generated by this investment has led to a bounty of potential applications for improvement of health, which are being explored for commercial purposes by the biotechnology industry. Private investment is thus ever present in development of these biotechnological products. All these factors raise important questions, including increasing concerns about intellectual property and conflict of interest. As this expansion continues, the established processes of peer review should be strengthened. PMID- 11728569 TI - Instrumental delivery and epidural anaesthesia. PMID- 11728570 TI - Instrumental delivery and epidural anaesthesia. PMID- 11728572 TI - Interstitial nephritis associated with celecoxib. PMID- 11728573 TI - Instrumental delivery and epidural anaesthesia. PMID- 11728574 TI - Treatment of neonatal hyperammonaemia. PMID- 11728576 TI - Antibiotics for adverse outcomes of pregnancy. PMID- 11728578 TI - Intestinal permeability in coeliac disease. PMID- 11728579 TI - Helicobacter pylori and reflux disease. PMID- 11728581 TI - Helicobacter pylori and reflux disease. PMID- 11728582 TI - Tick-borne encephalitis in Europe. PMID- 11728584 TI - Dichlorodiphenylethylene serum concentrations and effect on birthweight. PMID- 11728585 TI - Challenge to law on right to die. PMID- 11728586 TI - Adjustment of Europe's drug regulation to public- health needs. PMID- 11728587 TI - Mobile phones, precautionary principle, and future research. PMID- 11728589 TI - Venous thromboembolic disease in pilots. PMID- 11728590 TI - Adjustment of Europe's drug regulation to public-health needs. PMID- 11728600 TI - Wrong? And so what! PMID- 11728602 TI - Synthesis and application of caged peptides and proteins. AB - Caged compounds have covalently attached groups that are rapidly cleaved upon exposure to UV light. Attachment of photolabile groups makes the molecule inert until photolysis releases it in its bioactive form. When caged compounds are applied to the experimental system in advance, the concentration jump of biologically active substances can be brought about immediately in a limited area upon irradiation with pulsed and focused UV light. Therefore, caged compounds of low molecular weight, which are commercially available, have been used effectively to study the mechanisms of temporal biological phenomena, such as muscle contraction, intracellular signaling, and neurotransmission. Because many proteins and peptides play important roles in these phenomena, their caged derivatives should serve as powerful tools to clarify complex biological systems. To prepare caged proteins and peptides, several groups have improved upon a chemical modification method, as well as developed two new methods: (1) nonsense codon suppression and (2) solid-phase peptide synthesis. In this review, we summarize recent advances made in the design, preparation, and application of caged peptides and proteins. PMID- 11728603 TI - Understanding airway wall remodeling in asthma: a basis for improvements in therapy? AB - Current strategies for the management of asthma focus on suppressing airway inflammation. Other characteristic features of human asthma, such as airway hyperreactivity and the structural changes collectively referred to as airway remodeling, are largely ignored in existing guidelines for monitoring the effectiveness of treatment. Evidence is accumulating that pharmacologic therapy targeting airway wall remodeling may be valuable in treating asthma. However, development of appropriate therapeutic agents will require a better understanding of the pathogenesis of remodeling, which appears to be regulated by a variety of cytokines and growth factors produced by inflammatory, epithelial, and stromal cells. Furthermore, testing the effectiveness of novel agents that specifically target the process of remodeling will require appropriate experimental models, but most currently available animal models of asthma have major limitations. A recently described murine model of chronic human asthma offers considerable potential for dissection of the mechanisms of airway wall remodeling, as well as for investigation of the therapeutic potential of drugs that can modulate chronic inflammation and remodeling. PMID- 11728604 TI - Gene therapy in vascular medicine: recent advances and future perspectives. AB - Gene therapy is emerging as a potential strategy for the treatment of cardiovascular diseases, such as restenosis after angioplasty, vascular bypass graft occlusion, and transplant coronary vasculopathy, for which no known effective therapy exists. The first human trial in cardiovascular disease was started in 1994 to treat peripheral vascular disease using vascular endothelial growth factor. In addition, therapeutic angiogenesis using the vascular endothelial growth factor gene was applied in the treatment of ischemic heart disease. The results from these clinical trials seem to exceed expectation. Improvement of clinical symptoms in peripheral arterial disease and ischemic heart disease has been reported. At least five different potent angiogenic growth factors have been tested in clinical trials to treat peripheral arterial disease or ischemic heart disease. In addition, another strategy for combating disease processes, to target the transcriptional process, has been tested in a human trial. Transfection of cis-element double-stranded oligodeoxynucleotides is an especially powerful tool in a new class of antigen strategies for gene therapy. Transfection of double-stranded oligodeoxynucleotides corresponding to the cis sequence will result in the attenuation of the authentic cis-trans interaction, leading to the removal of trans-factors from the endogenous cis-elements, with subsequent modulation of gene expression. Genetically modified vein grafts transfected with a decoy against E2F, an essential transcription factor in cell cycle progression, revealed apparent long-term potency in human patients. This review focuses on the future potential of gene therapy for the treatment of cardiovascular disease. PMID- 11728605 TI - Genome analysis with gene-indexing databases. AB - The recent release of the draft sequence and the eventual completion of the human genome present the scientific community with a rich source of data to mine. Yet, these data are content poor in the absence of additional correlative information. Expressed sequence tag (EST) datasets and their associated gene indices have existed for many years, and represent the first attempt at understanding the complexity of the genome. These datasets remain extremely important as information sources and, in particular, as tools for analyzing the completed genomes. Here, we discuss the nature of ESTs and their associated tools and gene indexing databases. In particular, we will compare three EST gene indices (UNIGENE, Merck Gene Index Version 2.0 and Doubletwist CAT), discuss how these gene indices are applied for both genome analysis and drug discovery, and demonstrate their importance as a complementary dataset to the annotated human genome. PMID- 11728606 TI - Pharmacology of adenosine receptors in the vasculature. AB - Adenosine is widely distributed in mammals. One of the primary roles of adenosine within the cardiovascular system is to directly control the functions of both cardiac and vascular tissues. Recently, there has been considerable interest in the subclassification of adenosine receptors. Characterization of a heterogeneous population of receptors for adenosine could provide an opportunity for the development of novel compounds of therapeutic value. Adenosine is released from cells as a result of metabolism, and its release can be increased dramatically from cells that are metabolically stressed. This implies that adenosine can be released from a variety of cells throughout the body, as a result of increased metabolic rates, in concentrations that can have a profound impact on blood vessel function and, consequently, blood flow. It is recognized that the actions of this nucleoside on the vasculature are most prominent when oxygen demand is high and there is a reduction in oxygen tension at the site in question. Therefore, it is not surprising that adenosine has been shown to be an important regulator of blood vessel tone under hypoxic conditions. Furthermore, the activation of adenosine receptors on blood vessels can result in relaxation and/or contractions. The nature of the response subsequent to the activation of adenosine receptors is primarily dependent on the type of blood vessel involved and basal tone. This review will focus on the characterization of subtypes of adenosine receptors in blood vessels, as well as the effect of the stimulation of adenosine receptors on the peripheral circulation. PMID- 11728607 TI - Mechanisms of angioplasty and stent restenosis: implications for design of rational therapy. AB - Restenosis after angioplasty or stenting remains the major limitation of both procedures. A vast array of drug therapies has been used to prevent restenosis, but they have proven to be predominantly unsuccessful. Recent trends in drug therapy have attempted to refine the molecular and biological targets of therapy, based on the assumption that a single biological process or molecule is critical to restenosis. In contrast, both stenting and brachytherapy, which are highly nonspecific, can successfully reduce restenosis after angioplasty or stenting, respectively. This review examines the biology of both angioplasty and stent stenosis, focussing on human studies. We also review the landmark human trials that have definitively proven successful therapies, such as stenting and brachytherapy. We suggest that the successful trials of stenting and brachytherapy and the failure of other treatments have highlighted the shortcomings of conventional animal models of arterial intervention, and gaps in our knowledge of human disease. In contrast to arguments advocating gene therapy, these studies suggest that the most likely successful drug therapy will have a wide therapeutic range, targeting as many of the components or biological processes contributing to restenosis as possible. PMID- 11728608 TI - No association between dopamine D(2) and D(4) receptor gene variants and antidepressant activity of two selective serotonin reuptake inhibitors. AB - The possible association of the dopamine receptor D(2) (Ser 311Cys) and D(4) exon 3 (48 base pair repeat) gene variants with the antidepressant activity of selective serotonin reuptake inhibitors (SSRIs) was investigated in a sample of 364 inpatients affected by a major depressive episode treated with fluvoxamine, 300 mg/day (n=266), or paroxetine, 20-40 mg/day (n=98). The severity of depressive symptoms was assessed weekly with the Hamilton Rating Scale for Depression. Dopamine receptor D(2) (DRD2) and dopamine receptor D(4) (DRD4) allelic variants were determined in each subject by polymerase chain reaction. We observed that DRD2 and DRD4 variants were not associated with response to SSRI treatment. Possible stratification factors, such as sex, diagnosis, presence of psychotic features, depressive symptoms at baseline, paroxetine and fluvoxamine plasma levels, and pindolol augmentation did not significantly influence the observed results. The investigated DRD2 and DRD4 gene variants therefore do not seem to play a major role in the antidepressant activity of SSRIs, at least in the present sample. PMID- 11728609 TI - Personality dimensions in pathological gambling disorder and obsessive-compulsive disorder. AB - This study was conducted to investigate the similarities and differences in the personality dimensions of patients with pathological gambling disorder (PGD) and obsessive-compulsive disorder (OCD). Thirty-three subjects with PGD, 41 with OCD and 40 normal controls were assessed with the Tridimensional Personality Questionnaire (TPQ), which assesses three personality dimensions: novelty seeking, reward dependence, and harm avoidance. Compared with OCD subjects, PGD subjects expressed significantly greater novelty seeking, impulsiveness, and extravagance. The PGD subjects also reported significantly less anticipatory worry, fear of uncertainty, and harm avoidance than the OCD subjects. Compared with controls, the PGD subjects expressed significantly greater novelty seeking, impulsiveness, and extravagance. These results suggest that the personality dimensions of pathological gamblers may differ significantly from both those of OCD patients and normal controls. PMID- 11728610 TI - Impairment of incidental memory for frequency in patients with obsessive compulsive disorder. AB - The aim of our study was to investigate whether patients with obsessive compulsive disorder (OCD) have impaired incidental memory for frequency. Fifty four subjects (27 OCD patients and 27 matched control subjects) performed a task assessing estimation of occurrence of previously heard words. Performance on this task was compared with other intentional verbal memory tasks (recognition, recall and learning of common words). We also correlated memory for frequency with frontal lobe tests (Wisconsin Card Sorting Test, Trail Making Tests A and B, and verbal fluency). Performance on incidental learning of frequency was significantly worse in the OCD group than in control subjects. Other verbal memory measures did not show significant differences. Performance in the frequency task correlated with verbal fluency. Although intentional verbal memories are normal in OCD patients, incidental memory for frequency is impaired, suggesting that prefrontal systems are involved in OCD. PMID- 11728611 TI - The use of antibody D8/17 to identify B cells in adults with obsessive-compulsive disorder. AB - Compared with healthy control subjects, individuals with childhood-onset obsessive-compulsive disorder (OCD) have been reported to have a higher percentage of B cells that react with the monoclonal antibody D8/17, a marker for rheumatic fever. This study sought to replicate these findings in adults with OCD. Double-blind analyses of blood samples from 29 consecutive adults with primary OCD and 26 healthy control subjects were conducted to determine the percentage of B cells identified by D8/17. Using a standard criterion of > or =12% labeled B cells to denote positivity, rates of D8/17 positive individuals did not significantly differ between the OCD (58.6%) and control (42.3%) groups. Early age of onset was not a predictor of D8/17 positivity in the OCD group. The percentage of B cells identified by the monoclonal antibody marker D8/17 did not distinguish adults with OCD from control subjects, nor did it distinguish a sub group of adults with OCD who described pre-pubertal onset of their OCD symptoms. PMID- 11728612 TI - Do panic patients process unconditioned fear vs. conditioned anxiety differently than normal subjects? AB - Panic patients were evaluated with two models of experimental anxiety that are believed to generate distinct emotional states: (1) a stimulated public speaking test (SPS), a presumed indicator of unconditioned fear, and (2) conditioning of skin conductance responses (CSCR) to a tone associated with an aversive white noise, an index of conditioned anxiety. Subjective states were evaluated through the visual analogue mood scale (VAMS) and a bodily symptoms scale (BSS). In the SPS test, panic patients showed higher baseline levels of VAMS-measured anxiety than controls. Unlike controls, panic patients failed to show increased anxiety before and during speech. Although baseline levels of arousal were similar in both groups, VAMS mental sedation decreased in controls, but not in panic patients during the SPS. Panic patients showed more discontent than controls throughout the whole experimental session. They also scored higher than controls on several items of the BSS. In the CSCR test, panic patients showed more spontaneous fluctuations of skin conductance than controls. Nevertheless, conditioning of skin conductance responses to the tone was similar in both groups. Therefore, panic patients seemed to process unconditioned fear abnormally. PMID- 11728613 TI - Dopaminergic augmentation of sleep deprivation effects in bipolar depression. AB - Total sleep deprivation (TSD) has been used in association with lithium salts and with serotonergic and noradrenergic antidepressants, leading to sustained improvements in patients affected by major depression. Current theories on the neurobiological mechanism of action of TSD propose a major role for enhanced dopamine activity. To test the clinical relevance of dopaminergic enhancement in TSD, we treated a homogeneous sample of 28 bipolar depressed patients with three cycles of TSD combined with placebo or with the dopaminergic antidepressant amineptine. Changes in mood over time were rated with self-administered visual analogue scales and with the Montgomery-Asberg Depression Rating Scale. Patients showed improved mean daily-mood scores after TSD, an effect that was highest at the first cycle and decreased with treatment repetition. Amineptine enhanced the effects of TSD on perceived mood during the first two TSD cycles, but patients in the placebo and amineptine groups showed comparable results at the end of the treatment. Despite its theoretical importance, the clinical usefulness of combining TSD with a dopaminergic agent must be questioned. PMID- 11728614 TI - Three-dimensional structure of the Hospital Anxiety and Depression Scale in a large French primary care population suffering from major depression. AB - Few studies have been specifically carried out to characterize the dimensional structure of the Hospital Anxiety and Depression Scale (HADS) and those that have, have yielded contradictory results. We have examined the factor structure and sensitivity to change of the HADS in a large French outpatient primary care population treated with sertraline for major depression (DSM-IV criteria). Factor analysis of the HADS was performed in 2669 outpatients and in subsamples using a principal component procedure with Varimax rotation. Concurrent change sensitivity of the HADS was compared with that for the Hamilton Depression Rating Scale (HDRS) after at least 45 days of sertraline treatment. Three distinct factors emerged from the HADS factor analysis: a "depression" factor and two separate anxiety subscales: "psychic anxiety" and "psychomotor agitation" whose mean reductions in scores from baseline were significantly correlated (0.36-0.45) with the reduction of the HDRS baseline score. These new data provide support for the use of the HADS's three-dimensional structure to measure improvement of selected symptoms of anxiety during antidepressant therapy. PMID- 11728615 TI - Characteristics of fluvoxamine-induced nausea. AB - We investigated the association between fluvoxamine and nausea from various viewpoints. The incidence of nausea induced by fluvoxamine was 29% (12/41). Plasma 5-hydroxyindoleacetic acid (p5-HIAA) levels after fluvoxamine administration were significantly higher in patients with nausea (6.6+/-3.4 ng/ml) than in those without nausea (3.5+/-2.7 ng/ml). On the other hand, no significant differences were found between patients with and patients without nausea in terms of sex, age, initial and maximum dosages of fluvoxamine and its plasma concentrations, and clinical response to fluvoxamine. However, the incidence of nausea in patients who were initially administered fluvoxamine at under 50 mg/day was significantly lower than in those who were started at above 50 mg/day. In addition, mosapride, a member of the benzamide family, was effective in alleviating fluvoxamine-induced nausea. These results suggest that fluvoxamine-induced nausea is associated with hyperactivity in serotonergic neurons. PMID- 11728616 TI - Effect of electroconvulsive therapy on hematological parameters. AB - Although a complete blood count is part of the evaluation before the use of electroconvulsive therapy (ECT), there are no known hematological contraindications for the procedure. A preliminary study was done on 31 randomly selected psychiatric patients (chronic schizophrenia, n=10; acute depression, n=8; acute mania, n=6; acute psychosis, n=6; delusional disorder, n=1) receiving ECT to study its hematological effects. Blood samples were drawn just before and 0, 1 and 2 h after ECT. Hemoglobin (Hb%), total and differential leukocyte count (TLC and DLC), red blood cell (RBC) count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and platelet count were measured on a fully automated hematology analyzer (Sysmex K-1000). Significant changes were found in TLC, percentage of polymorphs and lymphocytes, and Hb%. Changes in other parameters were not statistically significant. More such studies are needed to substantiate these observations and to understand the mechanism and implication of these effects. PMID- 11728617 TI - The Asthma Clinical Research Network. PMID- 11728618 TI - The organization of the Asthma Clinical Research Network: a multicenter, multiprotocol clinical trials team. AB - Asthma is an increasingly serious cause of morbidity and mortality in the United States, affecting approximately 12 million people, including men and women, children and adults, and all racial and ethnic groups. It is now recognized that asthma is a complex disease of varied etiology triggered by a number of factors such as allergens, drugs, chemicals, exercise, cold dry air, infections, and emotions. Asthma is a chronic disease requiring multiple medications to treat and control symptoms as well as medications thought to control the underlying inflammation. Despite major advances in understanding the etiology and pathophysiology of asthma and the development of new therapeutic modalities, the prevalence, severity, and mortality from asthma have all increased over the past decades in all age groups. Hospitalizations for asthma have doubled in adults and increased fivefold for children over the past 20 years. Mortality appears to be particularly high in urban and rural minority populations. Asthma continues to place a heavy burden on patients and their families as well as the health-care system. In an attempt to respond to the need for well-designed clinical trials to allow rapid evaluation of new and existing therapeutic approaches for asthma and for dissemination of laboratory and clinical findings to the health-care community, the Division of Lung Diseases, National Heart, Lung, and Blood Institute, established the Asthma Clinical Research Network. PMID- 11728619 TI - The clinical trials in the initial five-year award period of the Asthma Clinical Research Network. AB - During its first years of existence, the Asthma Clinical Research Network initiated four major clinical trials and one pilot clinical trial. The objective of this article is to describe briefly the specific aims, design, and conduct of those five trials. PMID- 11728620 TI - Development and deployment of an internet-based data management system for use by the Asthma Clinical Research Network. AB - Data management system development for the first Asthma Clinical Research Network (ACRN) study began at the data coordinating center (DCC) in May 1995 with the requirement for delivery of a production system by November 1995. Special methods had to be used to establish an internet local area network (LAN), place clinical client systems, and achieve an accelerated software development cycle. The development of a fully integrated data management system prior to the start of the study was not possible. Therefore an early analysis focused on identifying discrete groupings of data management functions that would allow development of distinct database modules to provide specific functionality such as subject randomization, subject registration, and data entry. The modules were categorized as either being associated with clinical centers or the DCC so that the clinical center modules could be developed and delivered to meet the start date of the study. In the second phase of development during the relatively slow patient enrollment period, the DCC functional modules were delivered discretely over time. While at the time this development model was a necessity due to limited DCC resources, it continues to be used today as it permits the DCC to implement studies more rapidly and efficiently for the ACRN. This paper describes the methodologies used to develop an internet-based LAN, establish clinical center client systems, establish DCC client and server operations, and develop a data management system. It describes the circumstances that contributed to the development of these systems and the special methodologies developed. The technical aspects of the data management system and LAN are presented as well as a description of the requirements and constraints analysis used to develop the hardware and software systems. PMID- 11728621 TI - Modem remote support of pulmonary-function testing and quality control systems. AB - The data coordinating center (DCC) of the Asthma Clinical Research Network (ACRN) is responsible for the support of 11 pulmonary-function testing systems and two quality control systems. Pulmonary-function data from these systems are used as outcome indicators in studies conducted by the ACRN. Each of these systems is composed of a spirometer, a personal computer for data acquisition from the spirometer, a modem, and a printer. These systems are located at six clinical centers nationwide. An analysis conducted at the beginning of the first ACRN protocol identified the following requirements: (1) standard pulmonary-function testing, (2) standard methacholine-challenge testing, (3) the ability to handle simultaneous multiple protocols as well as have data from non-ACRN subjects, (4) the ability to separate data from different protocols as well as separate ACRN and non-ACRN data, (5) the ability to transmit data from the remote clinical centers to the DCC, (6) the ability to ensure quality data and to report on those results, and (7) the ability to provide remote support. PMID- 11728622 TI - Data management procedures in the Asthma Clinical Research Network. AB - Well-designed data management processes are essential in ensuring the quality of data collected in multicenter clinical trials. This paper describes the data management processes and systems that were developed by the data coordinating center of the Asthma Clinical Research Network. A combination of manual and electronic processes has been designed to process clinical trial data from the point of collection to statistical analysis. A distributed database management system consisting of modular applications for separate data processing activities was developed to enter, track, verify, validate, and edit collected data. In addition, processes for monitoring and reporting data quality are discussed. PMID- 11728623 TI - Implementing randomization procedures in the asthma clinical research network. AB - Randomization is a required component for the success of most controlled clinical trials. To ensure that the benefits of randomization are realized, well defined and carefully planned procedures must be put in place prior to the start of a trial. This paper presents a detailed account of the registration and randomization procedures implemented for the first four clinical trials of the Asthma Clinical Research Network. PMID- 11728624 TI - Genetic analysis in the Asthma Clinical Research Network. AB - Because there is reason to believe that genetic variants could account for different treatment responses in subjects with asthma, it is important to collect blood for genetic-analysis purposes when conducting clinical trials in asthma. This article describes issues related to maintaining subject confidentiality, tracking and shipping blood samples, quality control procedures at the laboratory performing the genotyping, and necessary data verification checks when implementing the genetic-analysis database for the Asthma Clinical Research Network. PMID- 11728625 TI - Quality control within the Asthma Clinical Research Network. AB - Quality control and assurance are very important issues for multicenter clinical investigation. Although such investigation usually states that those factors are part of the investigation, at best this occurs to a minimal extent. Indeed, to ensure the best possible investigatory results with the least amount of variance between centers, the Asthma Clinical Research Network (ACRN) instituted strict written criteria that were continually overviewed. This article describes the ACRN's quality control of: clinic procedures and equipment, training and certification of study personnel, site visit evaluations, and the procedures of bronchoscopy and sputum induction. The examples given in this report demonstrate how various aspects of our investigation are performed. PMID- 11728626 TI - Recruitment strategies in the Asthma Clinical Research Network. AB - In any clinical trial, recruitment of trial participants usually requires an intense effort to reach the target sample size. The Asthma Clinical Research Network (ACRN) was keenly aware of this issue at its inception and therefore initiated and emphasized recruitment strategies for each of its clinical trials. This article describes the recruitment strategies for the ACRN's first four major clinical trials. Particular attention is given to the strategies for the recruitment of women and minorities. Finally, the specific strategies of each of the six ACRN clinical centers are presented. PMID- 11728627 TI - Attrition and retention of ethnically diverse subjects in a multicenter randomized controlled research trial. AB - Recruitment and retention of subjects is critically important for the conduct of randomized clinical trials. When significant effort is given toward recruiting members of ethnic minorities in a sample, a similar rate of retention for all subgroups of subjects is essential. Little is known about why research subjects withdraw consent to participate in research and whether attrition is influenced by ethnicity or other factors. The purpose of this study was to determine the reasons for withdrawal from a large, multicenter randomized trial and whether these reasons differed among ethnic and gender subgroups. Additionally, we were interested in comparing withdrawn subjects to matched subjects who stayed in the trial to determine what factors encouraged full study completion. Using a cohort survey design, adult subjects (n = 35) who withdrew from a large, multicenter randomized trial and matched subjects who completed the study (n = 35) were interviewed by telephone after study completion. Subjects who withdrew consent tended to be female and members of ethnic minorities. The most frequent problems reported significantly more often by subjects who withdrew consent compared to matched retained subjects-were interference with work, lack of time, complicated and cumbersome record-keeping requirements, difficult study medicine regimens, and difficulty rescheduling appointments due to lack of flexibility on the part of study personnel. Only 17% of withdrawn subjects were satisfied with the overall research experience compared to 52% of matched retained subjects. Matched subjects who completed all required visits in the randomized controlled trial reported three reasons that promoted completion: remuneration, commitment to finish, and belief that the study was important. PMID- 11728628 TI - Summarizing methacholine challenges in clinical research. AB - In clinical trials in asthma, airway reactivity is commonly assessed by performing a methacholine challenge. Airway reactivity is thought to vary in proportion to asthma severity, and methacholine causes the airways of asthma subjects to constrict, thus lowering forced expiratory volume in 1 second (FEV(1)). A dose-response curve is obtained for each subject who meets standardized eligibility requirements to proceed with a methacholine challenge. When data from a methacholine challenge are used as an outcome variable in analysis, a univariate measure called the PC(20), the concentration of methacholine needed to produce a 20% fall in FEV(1) from baseline, is typically used to summarize the dose-response curve. Questions that arise regarding data generated from the methacholine challenge include: how to express data that do not yield a PC(20) value; whether PC(20) actually represents the best way to capture airway activity as expressed in a methacholine challenge; and whether the baseline FEV(1) is defined appropriately in calculation of PC(20). The impact of these issues on the statistical analysis of methacholine challenge data is described in this article. Some adjustments to the usual estimates of PC(20) and parametric modeling of the entire dose-response curve are proposed as alternatives that address some of the shortcomings of PC(20). PMID- 11728629 TI - Protection of translation initiation factor eIF2 phosphorylation correlates with eIF2-associated glycoprotein p67 levels and requires the lysine-rich domain I of p67. AB - The rate of protein synthesis in mammals is largely regulated by phosphorylation of the alpha-subunit of eukaryotic initiation factor 2 (eIF2) that is modulated by the cellular glycoprotein, p67, due to its protection of eIF2alpha phosphorylation (POEP) activity. At the N-terminus of p67, there are three unique domains, and at the C-terminus there is a conserved amino acid sequence. To analyze the importance of these domains, C-terminal deletion mutants of rat p67 were expressed constitutively in KRC-7 cells. In these cells, the phosphorylation level of the alpha-subunit of eIF2 was determined, and it was found that expression of the 1-97 amino acid segment of rat p67 increases POEP activity in vivo, and induces the endogenous levels of p67. These cells also show increased growth rate, and efficient translation of chloramphenicol acetyltransferase and beta-galactosidase reporter genes. At the N-terminus of p67, there are two unique domains: a lysine-rich domain I with the sequence (36)KKKRRKKKK(44), and an acidic residue-rich domain with the sequence (77)EEKEKDDDDEDGDGD(91). Substitution of lysine-rich domain I with (36)NMKSGNKTQ(44) in rat recombinant p67 resulted in the inhibition of its POEP activity, and substitution of the acidic residue-rich domain with (77)QNIQKALEPEAGDGA(91), resulted in no inhibition of POEP activity in KRC-7 cells. Taken together, our data suggest that protection of translation initiation factor eIF2 phosphorylation correlates with eIF2-associated glycoprotein p67 levels and requires the lysine-rich domain I of p67. PMID- 11728630 TI - A single element in the phosphoenolpyruvate carboxykinase gene mediates thiazolidinedione action specifically in adipocytes. AB - Phosphoenolpyruvate carboxykinase (PEPCK) is the key enzyme in glyceroneogenesis, an important metabolic pathway that functions to restrain the release of non esterified fatty acids (NEFAs) from adipocytes. The antidiabetic drugs known as thiazolidinediones (TZDs) are thought to achieve some of their benefits by lowering elevated plasma NEFAs. Moreover, peroxisome proliferator activated receptor gamma (PPARgamma) mediates the antidiabetic effects of TZDs, though many TZD responses appear to occur via PPARgamma-independent pathways. PPARgamma is required for adipocyte PEPCK expression, hence PEPCK could be a major target gene for the antidiabetic actions of TZDs. Here we used tissue culture and transfection assays to confirm that the TZD, rosiglitazone, stimulates PEPCK gene transcription specifically in adipocytes. We made the novel observation that this effect was by far the most rapid and robust among several other genes expressed in adipocytes. Adipocytes were transfected with a PEPCK/chloramphenicol acetyltransferase chimeric gene, in which either of the two previously discovered PPARgamma/retinoid X receptor alpha response elements, PCK2 and gAF1/PCK1, had been inactivated by mutagenesis. We demonstrate that PCK2 alone is a bona fide thiazolidinedione response element. We show also that the regulation of PEPCK by PPARs is cell-specific and isotype-specific since rosiglitazone induces PEPCK gene expression selectively in adipocytes, and PPARalpha- and PPARbeta-specific activators are inefficient. Hence, TZDs could lower plasma NEFAs via PPARgamma and PEPCK by enhancing adipocyte glyceroneogenesis. PMID- 11728631 TI - A high density lipoprotein from Piaractus mesopotamicus, pacu, (Osteichthyes, Characidae), is associated with paraoxonase activity. AB - We have characterized the serum lipoprotein profile and localized the serum paraoxonase activity of pacu, Piaractus mesopotamicus, a tropical fish species. The total lipoprotein profile of pacu serum obtained after KBr density ultracentrifugation shows the predominance of HDL (1.1267 g/mL). SDS-PAGE electrophoresis revealed a negligible amount of LDL. Pacu HDL was purified by gel filtration column on HPLC, and its molecular mass was estimated to be 246 kDa. Protein composition was 35%, and comprised four protein components with molecular masses of 45, 38, 25 and 12.5 kDa. Lipids represent 58% of total HDL, comprising 40% neutral lipids and 18% phospholipids by weight. The HDL contains 7% of carbohydrates, and mannose was the only sugar detected by paper chromatography in HDL hydrolysates. HDL-containing fractions showed the major paraoxonase activity. Purification of HDL resulted in a 23-fold enrichment of this activity. This is the first experimental evidence demonstrating the association of paraoxonase activity with a HDL in fish. PMID- 11728632 TI - Dissociation and unfolding of GCN4 leucine zipper in the presence of sodium dodecyl sulfate. AB - The dissociation and unfolding behavior of the GCN4 leucine zipper has been studied using SDS titration. Circular dichroism (CD) spectra showed that the alpha-helix content of the leucine zipper (20 microM) decreased during the sodium dodecyl sulfate (SDS) titration. However, the alpha-helix content of the leucine zipper still remained significant in the presence of 1 mM SDS, with little change detected when the SDS concentration further increased to 2 mM. The dimer dissociation of the leucine zipper is also a co-operative process during SDS titration; with no dimer remaining when SDS concentration reached 1 mM, as shown by electrophoresis and the the theta(222)/theta(208) ratio. Our results indicate that SDS efficiently induces leucine zipper dimer dissociation with the monomers still partially folded. The experimental results provide important evidence for the previous model that partial helix formation precedes dimerization in coiled coil folding. PMID- 11728633 TI - Phosphatidylserine increase in rat liver endomembranes during the acute phase response. AB - The acute phase response (APR) is responsible for great changes in protein and lipid metabolism. For example, marked changes are observed in the metabolism of fatty acids, triglycerides, cholesterol and sphingolipids. Those lipids are partly recovered in the lipoproteins and subsequently in the plasma. Beside these lipid families, nothing is known about phospholipids and their synthesis in endomembranes during the APR. Our studies show that phosphatidylserine synthesis is stimulated during the APR and that this lipid is increased in the endoplasmic reticulum (ER) and the ER-derived vesicles. PMID- 11728634 TI - Chemical mechanism of beta-xylosidase from Trichoderma reesei QM 9414: pH dependence of kinetic parameters. AB - The variation of kinetic parameters of beta-xylosidase from Trichoderma reesei QM 9414 with pH was used to elucidate the chemical mechanism of the p-nitrophenyl beta-D-xylopyranoside hydrolysis. The pH-dependence of V and V/K(m) showed that a group on the enzyme with a pK value of 3.20 must be unprotonated and a group with a pK value of 5.20 must be protonated for activity and both are involved in catalysis. Solvent-perturbation studies indicated that these groups are neutral acid type. Temperature dependence of kinetic parameters suggested the stickiness of the substrate at lower temperatures than the optimum and the calculated ionization enthalpies pointed to carboxyl groups as responsible for both pKs. Chemical modification with triethyloxonium tetrafluoroborate and protection with the substrate studies demonstrated essential carboxyl groups on the enzyme. Profiles of pK(i) for D-gluconic acid lactone indicated that a group with a pK value of 3.45 must be protonated for binding and it has been assigned to the carboxyl group of D-gluconic acid formed by lactone ring breakdown in solution. PMID- 11728635 TI - JAB1/CSN5 interacts with the GAL4 DNA binding domain: a note of caution about two hybrid interactions. AB - The Jun activation domain binding protein 1 (JAB1) was first identified as an interaction partner and coactivator of c-Jun. Subsequently, it was found to be a subunit of the COP9 signalosome (CSN) and termed CSN subunit 5 (CSN5). This complex regulates light-mediated development in plants and plays an essential role in a variety of organisms. A striking feature of JAB1/CSN5 is its reported interaction with a wide range of proteins and its modulation of their activity or stability. We applied the yeast two-hybrid system to screen for proteins interacting with the DNA-binding domain of the transcription factor c-Myb and found JAB1/CSN5 among the double-positive clones. To our surprise JAB1/CSN5 was shown to interact with the DNA-binding domain of GAL4 alone and had to be rejected as a false positive in the GAL4-based two-hybrid system. This finding emphasizes the necessity of particular caution when JAB1/CSN5 is found in two hybrid screenings. PMID- 11728636 TI - Keratan sulphate in cerebrum, cerebellum and brainstem of sheep brain. AB - Keratan sulphate was identified in sheep brain. We describe here the isolation and partial characterization of keratan sulphate from cerebrum, cerebellum and brainstem of young sheep brains. The galactosaminoglycan was isolated by using ion-exchange chromatography and gel filtration after exhaustive digestion with papain of the delipidated tissues, followed by alkaline borohydride degradation and chondroitinase ABC and heparinases I, II and III treatment. The material isolated by ion-exchange chromatography from each tissue was eluted as single but polydispersed peak from Sephadex G-75, with average molecular masses 8.4, 7.9 and 8.8 kDa for cerebrum, cerebellum and brainstem, respectively. Keratanase I and II totally degraded keratan sulphate from cerebrum and brainstem, but only partially that from cerebellum. The content of keratan sulphate was found to be about 215, 173 and 144 microg/g dry delipidated tissue for cerebrum, brainstem and cerebellum, respectively. PMID- 11728637 TI - Purification and characterization of phosphoglucose isomerase allozymes from Daphnia magna. AB - Phosphoglucose isomerase (PGI, EC 5.3.1.9) is polymorphic in many populations. Frequently, it has been shown that naturally occurring allozymes exhibit strong deviations form Hardy-Weinberg expectations, suggesting fitness relevant mutations. To investigate the nature of this allozymic variation, PGI was purified from Daphnia magna to high purity yielding a specific activity of 135.2 U/mg. The kinetic parameters of the allozymes were characterized depending upon ionic strength, pH and viscosity. The half-saturation constants of the allozymes were all equal, while the specific activity of the PGI from heterozygotes was consistently higher than the PGI of the homozygotes, independent of pH, ionic strength and viscosity of the solution. PMID- 11728643 TI - Perception and memory in neuroscience: a conceptual analysis. AB - Neuroscientists, in the last half of the 20th century, provided major insights into the cellular and molecular mechanisms associated with seeing and remembering. We first identify some of the most important of these discoveries. This is done along lines familiar to neuroscientists who have read many of the recent books and reviews that provide an overview of neuroscientific discoveries. In general, these emphasize the scientific contributions this discipline has made to our understanding of the mechanisms that give rise to the psychological attributes of humans and other animals. In the next sections, we examine the claims made in these overviews; in particular, those by the standard-bearers of neuroscience, in an attempt to clarify what can and what cannot be justified in these claims. This requires a conceptual analysis of a kind that is unfamiliar to most neuroscientists. Our analysis begins with consideration of the conceptual confusions that ensue when neuroscientists attribute seeing, remembering and other psychological attributes to the brain rather than to the creature whose brain it is. Subsequently, we outline what we take to be the appropriate conceptual scheme for neuroscientists to adopt. PMID- 11728644 TI - Spike timing, synchronization and information processing on the sensory side of the central nervous system. AB - To what extent is the variability of the neuronal responses compatible with the use of spike timing for sensory information processing by the central nervous system? In reviewing the state of the art of this question, I first analyze the characteristics of this variability with its three elements: synaptic noise, impact of ongoing activity and possible fluctuations in evoked responses. I then review the recent literature on the various sensory modalities: somato-sensory, olfactory, gustatory and visual and auditory processing. I emphasize that the conditions in which precise timing, at the millisecond level, is usually obtained, are conditions that usually require dynamic stimulation or sharp changes in the stimuli. By contrast, situations in which stimulation not belonging to the temporal domain is temporally encoded lead to much coarser temporal coding; although in both cases, neural networks transmit the signals with similarly high precision. Synchronization among neurons is an important tool in information processing in both cases but again seems to act either at millisecond or tens of millisecond levels. Information theory applied to both situations confirms that the average rate of information transmission is much higher in dynamic than in static situations. These facts suggest that channels of precise temporal encoding may exist in the brain but imply populations of neurons working in a yet to be discovered way. PMID- 11728645 TI - Cell signalling cascades regulating neuronal growth-promoting and inhibitory cues. AB - During development of the nervous system, neurons extend axons over considerable distances in a highly stereospecific fashion in order to innervate their targets in an appropriate manner. This involves the recognition, by the axonal growth cone, of guidance cues that determine the pathway taken by the axons. These guidance cues can act to promote and/or repel growth cone advance, and they can act either locally or at a distance from their place of synthesis. The directed growth of axons is partly governed by cell adhesion molecules (CAMs) on the neuronal growth cone that bind to CAMs on the surface of other axons or non neuronal cells. In vitro assays have established the importance of the CAMs (N CAM, N-cadherin and the L1 glycoprotein) in promoting axonal growth over cells, such as Schwann cells, astrocytes and muscle cells. Strong evidence now exists implicating the fibroblast growth factor receptor tyrosine kinase as the primary signal transduction molecule in the CAM pathway. Cell adhesion molecules are important constituents of synapses, and CAMs appear to play important and diverse roles in regulating synaptic plasticity associated with learning and memory. Negative extracellular signals which physically direct neurite growth have also been described. The latter include the neuronal growth inhibitory proteins Nogo and myelin-associated glycoprotein, as well as the growth cone collapsing Semaphorins/neuropilins. Although less well characterised, evidence is now beginning to emerge describing a role for Rho kinase-mediated signalling in inhibition of neurite outgrowth. This review focuses on some of the major themes and ideas associated with this fast-moving field of neuroscience. PMID- 11728647 TI - Functional ovarian cysts and the need to remove them. AB - OBJECTIVE: To review the need for the removal of functional ovarian cysts. MATERIALS AND METHODS: Over a 5-year period, between July 1994 and June 1999, all functional ovarian cysts files were reviewed in order to determine the main surgical indication. RESULTS: Thirty-four functional ovarian cysts (11.45%) were removed during that period. In 14 cases, the surgical operation was considered justified. In 32 cases, the cysts were not organic as supposed to be and had no reason to be removed. DISCUSSION: Reported functional ovarian cyst removal rates range from 15 to 30%. Present exploration means (particularly ultrasound-guided needle aspiration) does not permit to discriminate functional from organic cysts in more than 70% of the cases. CONCLUSION: It is impossible to reduce functional cyst removal rate to 0% but we should all evaluate our activity yearly and be able to limit this rate to less than 30%. PMID- 11728648 TI - Long-term outcome in term breech infants with low Apgar score--a population-based follow-up. AB - OBJECTIVE: To investigate the relation between low Apgar score in breech infants and handicap in childhood. STUDY DESIGN: A case-control study. A questionnaire to mothers of 323 non-malformed, singleton infants delivered in breech presentation at term, 105 cases with Apgar score below 7 at 5 min and 218 controls. RESULTS: Four cases (4.6%) and one control (0.5%) had cerebral palsy. In infants without cerebral palsy, speech/language problems were more frequent than controls (10.6 versus 3.2%) (P=0.02). There were no differences in rates of deficits in attention, motor control and perception (DAMP), epilepsy, cognitive developmental delay or learning disabilities. Absence of any handicap or disability was reported in 65 cases (75%) compared to 172 controls (92%) (OR: 3.9; 95% CI: 1.9 7.9). CONCLUSION: Even though low Apgar score indicates an increased risk of neurological sequelae, most (75%) breech infants with low Apgar score are without a handicap/disability at follow-up. PMID- 11728649 TI - Sero-prevalence and mother-to-infant transmission of hepatitis E virus among pregnant women in the United Arab Emirates. AB - OBJECTIVES: (1) To study the prevalence of HEV infection in a cohort of pregnant women in the United Arab Emirates (UAE). (2) The rate of mother-to-infant transmission. METHOD: Sera of 469 pregnant women residing in the UAE was tested for anti-HEV with ELISA and for HEV-RNA by polymerase chain reaction (PCR). Neonatal cord-blood and infant blood was similarly tested for anti-HEV and HEV RNA. Out of these 469 women, 33% were UAE nationals (Group A) and Egyptians (Group B), respectively, and 34% were from the Indian sub-continent (Group C). RESULTS: Out of 469 mothers 93 (20%) were anti-HEV positive and 28 (30%) of these 93 were HEV-RNA positive and symptomatic with on-going infection. The prevalence of anti-HEV was significantly increased amongst Groups B and C (Group A:B, P<0.001; Group A:C, P<0.001). Twelve of these 28 symptomatic women developed acute hepatic disease and two died undelivered, thus, were excluded from further study. Six of the remaining 10 women had non-fulminant acute viral hepatitis and four developed fulminant hepatic failure, of which one died in the early post partum period. The remaining 16 women had mild symptomatic disease with complete recovery as did the remaining nine women with acute illness. Of the 10 babies born to mothers with acute disease, two were pre-term, three had anicteric hepatitis and two babies were born with hypothermia and hypoglycaemia and died within 48h. Noteworthy is the fact that, the 26 infants born to the 26 HEV-RNA positive mothers developed acute/on-going clinical infection and were HEV-RNA positive. Apart from the two early neonatal deaths, the remaining 24 infants had full recovery. No infant of the other mothers were symptomatic or had HEV-RNA. Thus, the mother-to-infant transmission was 100%. The remaining 65 sero-positive women remained asymptomatic with no evidence of clinical infection, continued to be HEV-RNA negative and delivered at term. Their infants had no evidence of infection and were sero-negative by 9 months of age, suggesting transplacental transmission of IgG antibodies. CONCLUSIONS: There is a high prevalence of anti HEV among pregnant women residing in the UAE. Twelve of infected women developed fulminant/acute hepatitis resulting in three fatalities. Evidence suggests significant vertical transmission of HEV among HEV-RNA positive mothers with appreciable perinatal morbidity and mortality. Excluding mothers with acute hepatic disease, Breast-feeding appears to be safe in HEV sero-positive mothers. PMID- 11728650 TI - Transvaginal cervicoisthmic cerclage as an alternative to the transabdominal technique. AB - OBJECTIVES: To analyse the foetal outcome after transvaginal cervicoisthmic cerclage, to determine whether it is a valid alternative to the reference transabdominal technique. STUDY DESIGN: Description of the vaginal operative technique, retrospective review of 20 cervicoisthmic cerclage patients from 1990 to 2000. RESULTS: In the curative group of women with a history of late pregnancy losses or premature deliveries and usually previous failed classical cervical cerclage, the foetal survival rate improved from 18% before cerclage to 79% after cerclage. No operative complications were reported. In the prophylactic group, typically characterised by the absence of the cervix as a consequence of surgery for invasive cervical cancers, the foetal survival rate was 83% after cerclage. CONCLUSIONS: The transvaginal cervicoisthmic cerclage allows a high foetal survival rate without complications in highly selected patients with poor obstetrical history. Because of its simplicity, this technique should replace the transabdominal route for surgeons experienced in vaginal surgery. PMID- 11728651 TI - Prepregnancy body mass index in non-diabetic women with and without shoulder dystocia. AB - OBJECTIVE: To investigate the distribution of prepregnancy body mass index (BMI) in non-diabetic women with and without shoulder dystocia. STUDY DESIGN: Cases were 142 non-diabetic women experiencing shoulder dystocia during the period from 1 January 1993 to 31 December 1999. Shoulder dystocia was defined as the impossibility of delivering the fetal shoulders by standard procedures. Controls were 142 women vaginally delivering during the same period without experiencing shoulder dystocia. Cases and controls were matched for parity (primi-/multipara) and birthweight (+/-250 g). Women with diabetes mellitus, gestational diabetes or a history of shoulder dystocia in a previous birth were excluded. The BMI and selected obstetric data were extracted from an internal database in the department. RESULTS: Delivery was performed using McRoberts maneuvre (42%), Woods screw (50%) or by primary delivery of the posterior arm (8%). Women experiencing shoulder dystocia had significantly more labor augmentation and more instrumental deliveries. No differences were shown in the prevalence of low Apgarscores. The proportion of children with Erbs palsy and clavicular fracture was very close to be significantly different in cases or controls. However, these data does not allow any conclusion. The distribution of BMI was equal in cases and controls. CONCLUSION: Non-diabetic women experiencing shoulder dystocia do not have a higher BMI than non-diabetic women delivering without this experience, given a fixed fetal weight. PMID- 11728652 TI - Influence of magnesium sulphate and isradipine on human placental cotyledon fetal vessels in vitro. AB - Increased systemic vascular resistance and blood pressure during pregnancy may be due to impaired prostacycline PGI(2)-thromboxan (TX) A(2) balance. The aim of the study was to compare the influence of magnesium sulphate and isradipine on the placental vascular resistance induced by stable thromboxan A(2) analogue U 46619 in experimental bilateral perfusion of the human placental cotyledon. The research used the experimental model described by Schneider et al. [Am. J. Obstet. Gynecol. 114 (1972) 822]. The control group consisted of six placental lobule perfusions lasting 120 min each. Constant increase in perfusion pressure of mean value 185% of the initial pressure was obtained from about the 60th min of the experiment and maintained till the end. Having obtained constant increase in perfusion pressure from the 60th min of the experiment, magnesium sulphate and isradipine were administered along with thromboxan A(2) analogue U 46619 into the fetal circulation. Both drugs lowered perfusion pressure, experimentally increased by thromboxan A(2) analogue U 46619. The onset of magnesium sulphate activity was quicker than isradipine. According to the results of our in vitro study, magnesium sulphate and isradipine may have a beneficial effect on vascular resistance of cotyledon vessels in PIH in vivo, too. PMID- 11728653 TI - The vasoconstrictor response of uterine and mesenteric resistance arteries is differentially altered in the course of pregnancy. AB - OBJECTIVES: The purpose of the present study was to test the hypothesis that pregnancy is associated with an attenuation of the vasoconstrictor response not only in the uterine, but also in the systemic circulation. Decreased vascular reactivity should be characterized by an early onset to account for the rapid fall in peripheral resistance which was observed during the first third of gestation. STUDY DESIGN: Dose-response curves for phenylephrine hydrochloride (PE), angiotensin II (ANG II), and endothelin 1 (ET 1) were recorded from isolated pressurized uterine and mesenteric arterioles. Vessels were obtained from virgin, early (day 7-9) pregnant (EP) and late (day 19-21) pregnant (LP) rats. RESULTS: (1) In uterine resistance arteries, the response to PE and ANG II decreased early, but for ANG II the reduction did not persist. ET 1 sensitivity was unchanged in early, and diminished in late gestation; (2) in mesenteric vessels, sensitivity to ET 1 was enhanced in early pregnancy and did not differ from the non-pregnant level in late gestation. Sensitivity to PE and ANG II was unchanged in early, and reduced in late pregnancy. CONCLUSIONS: The vasoconstrictor response is not uniformly blunted during pregnancy. Changes in vascular reactivity are differentially regulated with respect to the agonist, to their time course, and. to the origin of the vessel. The relative increase of vasoconstrictor sensitivity in the splanchnic circulation during early pregnancy may prevent hypotensive dysregulation, while concentrations of endogenous vasodilators rise and the vascular filling state normalizes only gradually. PMID- 11728654 TI - A randomised trial of an intervention package designed to promote external cephalic version at term. AB - OBJECTIVE: To evaluate the effectiveness of an intervention package to promote external cephalic version (ECV). STUDY DESIGN: (1) Design and Setting--A randomised-controlled trial of 20 consultant-based maternity units in the West Midlands NHS Region. (2) Intervention Units--One consultant obstetrician from each unit was encouraged to promote changes in clinical practice within their unit. A multifaceted package including a workshop, written material, guidelines and videos was delivered to these consultants. (3) Control Units--No intervention. (4) Main Outcome Measure--The percentage of women with breech presentation at term who were offered ECV in the antenatal clinic, before and after the intervention. RESULTS: Pre-intervention, there was no difference between the intervention and control units in the proportion of women offered ECV (20 and 19%, respectively). Post-intervention, the proportions were 15% in the control group and 36% in the intervention group (P=0.016). CONCLUSION: This type of intervention package can alter clinical practice and increase the proportion of women with breech presentation at term being offered ECV after the intervention. PMID- 11728655 TI - Occupational physical activity and pregnancy outcome. AB - OBJECTIVE: To determine the effect of occupational physical activities on the outcome of pregnancy. METHODS: A case control study included two groups; the first one included 562 patients with spontaneous abortion as a case group, while the second one included 1200 ladies with full term deliveries as a matched control group. Occupational physical activity has been evaluated through the energy expenditure (fatigue score and intensity score) and the biomechanic load (abdominal pressure intensity score, relative chronic pressure and peak pressure scores) in both groups. RESULTS: The group of spontaneous abortion had a significant higher intensity score, fatigue score, unfavourable working hours and high working speed than the group of favourable pregnancy outcome. The study showed a significantly higher biomechanic load among the group of spontaneous abortion represented by higher peak pressure score, chronic pressure score, lifting and bending activities. The group of spontaneous abortion had a significant (P<0.05) higher housekeeping working hours per day than that of the control group. Also they had a significant larger family size and had a significant lower helping in housekeeping activities than the group of favourable outcome. CONCLUSION: Excessive occupational physical activity has a definite detrimental effect on the outcome of pregnancy. PMID- 11728656 TI - Gestational diabetes complicated by hydramnios was not associated with increased risk of perinatal morbidity and mortality. AB - OBJECTIVE: To determine whether gestational diabetes (GDM) complicated with hydramnios is associated with higher rates of perinatal morbidity and mortality than those with normal amniotic fluid (AFI). STUDY DESIGN: A case control study of 368 pregnant women with GDM was conducted between January 1995 and December 1996. The study group consisted of 184 women with hydramnios (AFI>25 cm) that were matched for maternal age, parity, and gestational age to 184 women with gestational diabetes and normal AFI. A Student's t-test, McNemar test and logistic regression test were used to compare the two groups. RESULTS: Both groups were under strict metabolic control; mean glucose levels of 5.2+/-2.4 mmol/l in the study group and 5.5+/-2.4 mmol/l in the controls (P>0.05). There were no significant differences between groups in the rates of severe preeclampsia, premature rupture of membranes (PROM), cesarean section rate, intrauterine growth restriction (IUGR), antepartum fetal death, fetal distress, prolapse of umbilical cord and placenta abruption. Even though the prevalence of large for gestational age (LGA) and birth weight were significantly higher in the study group (31 versus 19% and 3441+/-477 versus 3232+/-554 g, respectively), the prevalence of neonatal trauma was the same in both groups (2.6%). There was no significant difference in the Apgar scores (1 and 5 min), newborn hypoglycemia, metabolic acidosis and hyperbilirubinemia. CONCLUSIONS: Hydramnios in women with GDM was not associated with increased risk of perinatal morbidity and mortality. PMID- 11728657 TI - Short-term outcome of newborn infants: spinal versus general anesthesia for elective cesarean section. A prospective randomized study. AB - OBJECTIVE: To compare general and spinal anesthesia with respect to the short term outcome of newborns born by elective cesarean deliveries. METHODS: Pregnant women admitted to our hospital from January 1999 to July 2000, for whom elective repeat cesareans were planned after 37 weeks gestation, were allocated randomly after their informed consent to spinal anesthesia or general anesthesia. Maternal age, gestational age, birth weight, Apgar's score, hospital stay duration, and duration of cesarean section time were all noted. The rate of the neonatal respiratory depression, perinatal asphyxia, and admittance to the neonatal intensive care unit of the infants were documented. We also studied arterial samples withdrawn from the cord for the pH, bicarbonate, PaO(2) (oxygen pressure, arterial), and PaCO(2) (carbon dioxide pressure, arterial). The serum levels of creatine kinase with myocardial-specific isoform, aspartate aminotransferase, alanine aminotransferase, and total cortisol levels of the newborns were measured and served in ruling out perinatal stress and in confirming the diagnosis of perinatal asphyxia (and of myocardial damage). Statistical analyses was performed with the use of an unpaired Student's t-test, Chi-square test, and a power calculation was done. RESULTS: From the randomly selected patients, we had 38 (45.2%) infants for general anesthesia and 46 (54.8%) for spinal anesthesia. None of our primary endpoints favored any of the study groups, and the clinical short term outcome of the infants was similar in the neonates born both by spinal and general anesthesia (P>0.05). The biochemical assays did not rule out or confirm any differences in the occurrence of perinatal stress (P>0.05). CONCLUSION: Anesthesia type does not seem to influence the short-term outcome of the newborn infants for the elective cesarean deliveries. We believe that both spinal and general anesthesia could be performed in elective term cesarean deliveries without any risk to the newborn infants. PMID- 11728658 TI - IGFBP-1 and glycodelin serum levels before and after hysterectomy with and without bilateral oophorectomy in premenopausal women. AB - OBJECTIVES: The aim of our study was to investigate the role of the intact endometrium and ovaries for serum levels of insulin-like growth factor binding protein-1 (IGFBP-1) and glycodelin. STUDY DESIGN: In 35 premenopausal patients with a planned hysterectomy, serum measurements of IGFBP-1 and glycodelin were done before surgery and 1, 3, 5, and 10 days after surgery. Patients were divided into three groups according to the kind and time of operation: (1) hysterectomy with bilateral adnexectomy in the luteal phase and (2) hysterectomy without adnexectomy in the follicular phase or (3) the luteal phase. RESULTS: IGFBP-1-we could not show any differences in IGFBP-1 serum levels before and after hysterectomy with or without bilateral oophorectomy. Glycodelin-hysterectomized and oophorectomized patients showed decreasing serum levels up to day 3. After day 5, circulating concentrations of glycodelin increased continuously but remained below pre-operative levels. In both non-adnexectomized groups we saw a reduction up to day 5 but a rise at day 10. None of the results reached statistical significance. CONCLUSION: The results indicate that endometrium and ovary are not the only sources of IGFBP-1 and glycodelin. PMID- 11728659 TI - Insulin resistance in polycystic ovary syndrome: hyperandrogenemia versus normoandrogenemia. AB - OBJECTIVE: The goal of this study was to evaluate the insulin resistance and glucose tolerance in hyperandrogenemic and normoandrogenemic groups of patients with polycystic ovary syndrome (PCOS). STUDY DESIGN: In this cross-sectional study, 17 hyperandrogenemic and 14 normoandrogenemic, age and weight-matched non obese women with PCOS were studied. All patients had clinical hyperandrogenism and chronic anovulation with polycystic ovaries on ultrasound. Insulin resistance and glucose tolerance were determined by measuring insulin and glucose concentrations following a 75 g oral glucose tolerance test (OGTT). Fasting glucose to insulin ratio (FG:I ratio), insulin area under the curve (AUC(insulin)) during OGTT, and homeostasis model assessment for insulin resistance (HOMA-IR) were calculated. RESULTS: Hyperandrogenemic group of patients had fasting hyperinsulinemia, lower FG:I ratio, higher AUC(insulin), and HOMA-IR compared with normoandrogenemic group. The differences between two groups were statistically significant. CONCLUSION: PCOS has variable biochemical features. Hyperandrogenemia associated with insulin resistance differs from normoandrogenemia in this syndrome. Fasting insulin concentrations, FG:I ratio, AUC(insulin), and HOMA-IR are convenient markers for determining insulin resistance in PCOS. PMID- 11728660 TI - Uterine cavity assessment in infertile women: comparison of transvaginal sonography and hysteroscopy. AB - OBJECTIVES: To evaluate the diagnostic accuracy of transvaginal sonography (TVS) in detecting uterine cavity abnormalities in infertile patients, with reference to hysteroscopy as the gold standard method. STUDY DESIGN: At the Institute of Obstetrics and Gynaecology, University of Bari, 134 infertile women were investigated with both TVS and hysteroscopy. In all cases endometrial biopsy was performed. Sensitivity, specificity, and positive and negative predictive values were calculated for TVS, considering hysteroscopic diagnosis as the gold standard. Statistical analysis was performed by using McNemar test. RESULTS: There was one failed insertion of hysteroscope. Hysteroscopy diagnosed pathological findings in 58 out of 133 cases (44%). TVS was in agreement with 50 of the 58 (86%) of the pathological findings diagnosed at hysteroscopy. As a test for the detection of uterine cavity abnormalities, TVS in comparison with hysteroscopy had 84.5% sensitivity and 98.7% specificity, 98.0% positive predictive value and 89.2% negative predictive value. CONCLUSIONS: TVS is able to diagnose polyps, septum and submucous fibroids with quite significant accuracy, while hysteroscopy is indicated for their confirmation and removal. Thus, hysteroscopy provides both diagnostic and therapeutic capabilities, but TVS permits to proceed directly to operative hysteroscopy avoiding the need for a separate diagnostic procedure. PMID- 11728661 TI - Is high-risk human papillomavirus infection associated with cervical intraepithelial neoplasia eliminated after conization by large-loop excision of the transformation zone? AB - OBJECTIVE: To investigate whether high-risk HPV infection associated with cervical intraepithelial neoplasia (CIN) was successfully eliminated after electrosurgical conization by large-loop excision of the transformation zone (LLETZ). STUDY DESIGN: 142 women, who were admitted for conization of CIN 1-3 were recruited into a prospective follow-up study of HPV infection, including cervical sampling for HPV DNA before, and then 3, 6 and 12 months after surgery. We examined whether there were any differences in the rate of HPV DNA positivity after LLETZ between specific risk groups, such as patients with primary (i.e. before surgical treatment) high-risk HPV infection, CIN of different grades, and positive margins. RESULTS: We did not detect statistically significant differences between specific risk groups. According to the assay used (hybrid capture II) at the last follow-up visit 94% of primarily infected patients were completely free from infection with high-risk HPV types, while 6% had persisting HPV infection. CONCLUSIONS: With a detection limit of 5000 genomes/ml HPV DNA the hybrid capture II results revealed, that after electrosurgical removal of CIN in 94% of patients testing positive for high-risk HPV DNA prior to surgery were negative 12 months post-surgery. PMID- 11728662 TI - Tamoxifen and ovarian cysts: a prospective study. AB - PURPOSE: To prospectively follow a group of women with breast cancer on tamoxifen for the development of ovarian cysts. METHODS: 72 women were followed every 6 months with pelvic examination and vaginal ultrasound. Chi square and Student's t test were used for statistical analysis. RESULTS: The duration of treatment was 31.5+/-20 months. The mean age was 51.2+/-9.8 years. 55.6% were post-menopausal. Out of 72 women, 18 (25%) developed ovarian cysts. The mean age of women who developed ovarian cysts was significantly lower than in those who did not (47.0+/ 7.0 and 52.5+/-10.2 years, respectively, P=0.03), however, the mean duration of treatment was not significantly different (33.3+/-17.4 and 29.3+/-20 months, respectively, P=0.45). Out of 32, 14 (43.8%) pre-menopausal and out of 40, 4 (10%) post-menopausal women developed ovarian cysts (P=0.003). They developed the cysts after an average duration of 33.3+/-18 and 50.7+/-6.2 months, respectively (P=0.7). The average diameter of the cysts was 2.8+/-1.2 cm. All cysts were simple except for one pre-menopausal women. All the cysts in post-menopausal women resolved spontaneously. One pre-menopausal patient had a multi-loculated cyst, was operated and had a serious cystadenoma. In nine patients, the cysts resolved spontaneously and in three after discontinuation of tamoxifen, and one patient was lost to follow-up. All cysts were asymptomatic. CONCLUSION: Ovarian cysts frequently develop in women with breast cancer on tamoxifen. The majority of the cysts resolve spontaneously, therefore an expectant management with follow up ultrasonography is recommended. PMID- 11728664 TI - A syndromal and an isolated form of uterine arteriovenous malformations: two case reports. AB - Uterine arteriovenous malformations are rare lesions with a considerable risk potential. Clinical presentation varies from no signs over various degrees of menorrhagia to massive life threatening vaginal bleeding. This is the first report of congenital uterine arteriovenous malformations in two patients with primary infertility. In one case, the uterine lesions were found in conjunction with other congenital malformations suggesting the diagnosis of hemihyperplasia/lipomatosis syndrome. Etiology, symptoms, diagnostic and therapeutic work-up are discussed; pathological findings are illustrated. PMID- 11728663 TI - Laparoscopic staging of endometrial cancer using laparosonic instruments: comparison with electrosurgery. AB - OBJECTIVE: To compare perioperative parameters in two groups of women with different laparoscopic operative techniques in surgical staging of endometrial cancer (EC). STUDY DESIGN: Thirty randomly allocated and laparoscopically treated women with EC. Fifteen patients were operated by electrosurgery, 15 patients by laparosonic operative technique. Differences between the two groups were determined by the Wilcoxon rank-sum test. Probability (P) of less than 0.05 was considered significant. SETTING: Department of Gynecology and Obstetrics, Endoscopic Training Center, Baby Friendly Hospital, Kladno, Czech Republic. RESULTS: Laparoscopy was successfully completed in 29 patients. Laparoscopy assisted surgical staging of EC was performed based on the tumor grade and the depth of myoinvasion. In both groups, in total 18 and 5 women underwent pelvic lymphadenectomy (PLN) and infra-aortic lymph node sampling (IALS), respectively. Three patients had metastases in pelvic lymph nodes. In the electrosurgical hemostasis and laparosonic group the mean total time required to finish the whole operative procedure were 132.1 and 138.3 min, respectively, with no statistically significant difference (P=0.96). There were no significant differences between the groups in any intraoperative or postoperative follow-up variables, except for the number of excised lymph nodes where the difference between electrosurgery and laparosonic group (12.7 versus 18) was statistically significant (P=0.05). In one patient with intraoperative venous bleeding the laparosonic hemostasis was ineffective (successful procedure rate 93.3%). One patient from the electrosurgery group was converted to laparotomy due to injury to the epigastric vessels. This complication had no connection with the surgical techniques studied. CONCLUSION: It is concluded that both operative technique variants in laparoscopy-assisted surgical staging appear to be feasible and effective for patients with EC. PMID- 11728665 TI - Synchronous vulvar and breast cancer. AB - Synchronous vulvar and breast cancer is rare. A 44-year-old women presented with a lesion in the right labia majora and right upper quadrant breast lump. After work-up, she underwent radical wide local vulvar excision and modified radical mastectomy with axillary lymph node dissection. The pathology of the vulva revealed moderately-differentiated squamous cell carcinoma and that of the breast infiltrating ductal carcinoma. Only two such cases have been previously reported: one was an elderly patient and the second a young patient with HIV infection. Our patient is a young and healthy woman making her presentation a unique and rare case. PMID- 11728666 TI - A urachal leiomyoma misdiagnosed as an ovarian tumor. AB - A case of leiomyoma of the urachus occurred in a 39-year-old woman with an abdominal mass. It was initially diagnosed as an ovarian tumor. PMID- 11728667 TI - A huge abdominal mass mimicking ovarian cancer: p53-negative but aneuploid myxoid leiomyosarcoma of the uterus. AB - OBJECTIVE: Less than 20 myxoid leiomyosarcoma cases were reported in literature. Since, these tumors are very rare and may exhibit highly malignant behavior despite their low mitotic index, clinical course and optimum type of therapy of myxoid variant of leiomyosarcoma were not well understood. The goal of this report is to contribute the better understanding of this rare type of tumor. METHODS: A 39-year-old woman presented with a huge abdominal cystic mass. Laparotomy was performed and frozen section diagnosis was low-grade uterine leiomyosarcoma. TAH-BSO, omentectomy, pelvic lymph node and peritoneal samplings were carried out. No chemotherapy was performed after surgical therapy. RESULTS: Final histopathological diagnosis was uterine myxoid leiomyosarcoma. The tumor was p53-negative and had aneuploid DNA content. The patient tolerated well the operation and she is alive and free of disease after 24 months of primary surgical treatment. CONCLUSION: Uterine myxoid leiomyosarcoma may present a huge abdominal cystic mass and can be treated successfully with surgery alone. PMID- 11728668 TI - Pregnancy complicated by Evan's syndrome. AB - Evan's syndrome was initially diagnosed in a 26-year-old pregnant patient. Following the introduction of high dose steroid therapy, the patient developed possible disseminated gonococcal infection which was followed by preterm labor and abruptio placentae. A cesarean delivery was performed at the 34th week of pregnancy following platelet infusion. While the infant survived, the mother had delayed postpartum hemorrhage. PMID- 11728669 TI - Inevitable hysterectomy despite conservative surgical management in advanced cervical pregnancy: a case report. AB - A 35-year-old woman was diagnosed to have cervical pregnancy of 10.2 weeks duration. Methotrexate therapy was not chosen due to the presence of active tuberculosis. Since the patient had gradually increasing vaginal hemorrhage, she was taken to the operating room. A dilatation and curettage could not control the hemorrhage. A laparotomy was performed and the uterine arteries were ligated. A cervical hysterotomy was also performed to evacuate the products of conception. There was active bleeding in the cervix although no products of conception were observed. Two purse string sutures were placed, but the persistence of profuse bleeding despite these measures necessitated total hysterectomy. PMID- 11728670 TI - Giant cell myocarditis: a fatal cause of dyspnea in pregnancy. AB - The clinical course of a pregnant patient, who presented with progressive dyspnea and heart failure is described. Despite intensive care and resuscitative efforts to mother and child, both expired. The autopsy revealed giant cell myocarditis in the mother. Giant cell myocarditis can affect pregnant patients and should therefore be considered in the differential diagnosis of progressive dyspnea. PMID- 11728671 TI - Amniocentesis and amnioinfusion during pregnancy. Report of four complicated cases. AB - We report two foetal complications after amnioinfusion with dye injection for evaluation of severe oligohydramnios in pregnancy. In the first case, an underskin colouring was to disappear within a few days. In the second case, an irreversible skin after-effect with muscular atrophy and lesion of the underskin tissues developed progressively, although the trained practitioner did not notice any particular resistance during the injection of the dye and the ultrasonographer did not point out that the foetus had been stuck by the needle. Both later reported cases concern two life threatening maternal complications due to amniocentesis for foetal karyotyping indicated by maternal age, a septic shock occurred 24h after entering the amniotic cavity, leading to a long stay in intensive care units. In one of those cases, the patient recovered from a cardiac arrest. We come to the conclusion that the essential continuous ultrasonographic monitoring cannot avoid all complications, some of which may lead to severe adverse foetal effects and others may need an intensive medical care for the mother, mainly after iatrogenic chorioamnionitis. Experience of the perinatologist remains an important factor to limit the complications without avoiding them completely. The indications must be carefully evaluated, the information given to the patients must be clear and in the future, intraamniotic injections of dyes will have to be avoided and replaced by non-invasive tests for diagnosis of preterm premature rupture of membranes, such as diamine-oxidase, foetal fibronectin or PROM-tests. The rules of a good practice will have to be defined and widely spread. PMID- 11728672 TI - Placenta percreta--two cases and review of the literature. AB - Two cases of placenta pecreta confirmed histologically were treated conservatively with retention of the uterus. Both later went on to have successful pregnancies. PMID- 11728673 TI - Re: Ledee et al. Management in intractable obstetric haemorrhage: an audit study on 61 cases. Eur J Obstet Gynecol Reprod Biol 2001;94:189-96. PMID- 11728675 TI - Personal thoughts on normal tissue tolerance, or, what the textbooks don't tell you. PMID- 11728676 TI - Sildenafil citrate (Viagra) and erectile dysfunction following external beam radiotherapy for prostate cancer: a randomized, double-blind, placebo-controlled, cross-over study. AB - PURPOSE: To determine the efficacy of sildenafil citrate (Viagra) in patients with erectile dysfunction after three-dimensional conformal external beam radiotherapy (3D-CRT) for prostate cancer. METHODS AND MATERIALS: 406 patients with complaints of erectile dysfunction and who completed radiation at least 6 months before the study were approached by mail. 3D-CRT had been delivered (mean dose 68 Gy). Sixty patients were included and entered a double-blind, placebo controlled, cross-over study lasting 12 weeks. They received during 2 weeks 50 mg of sildenafil or placebo; at Week 2 the dose was increased to 100 mg in case of unsatisfactory erectile response. At Week 6, patients crossed over to the alternative treatment. Data were collected using the International Index of Erectile Function (IIEF) questionnaire, and side effects were recorded. RESULTS: Mean age was 68 years. All patients completed the study. For most questions of the IIEF questionnaire there was a significant increase in mean scores from baseline with sildenafil, but not with placebo. Ninety percent of the patients needed a dose adjustment to 100 mg sildenafil. Side effects were mild or moderate. CONCLUSION: Sildenafil is well tolerated and effective in improving erectile function of patients with ED after 3D-CRT for prostate cancer. PMID- 11728677 TI - Can extraprostatic extension be treated by prostate brachytherapy? An analysis based on postimplant dosimetry. AB - PURPOSE: To determine whether extraprostatic extension (EPE) can be treated by Pd 103 prostate implants. METHODS AND MATERIALS: The postimplant dosimetry of 22 consecutive Pd-103 prostate implants was analyzed to determine whether potential EPE was adequately treated. The implants were peripherally loaded and planned with a 3-5-mm dose margin at midgland. Seeds were not implanted outside of the capsule except at the base and apex. The postimplant dosimetry was based on a CT scan obtained 32 +/- 8 days postimplant. The radial distance between the prostate edge and the prescription isodose line was measured at the left lateral, left posterolateral, posterior, right posterolateral, and right lateral positions on each prostate contour. Similar measurements were made of the preplan dose margins. RESULTS: The mean postimplant dose margin was > or =4.5 mm at the midgland and apex of the prostate in agreement with the preplan. However, at the base, the mean margins at the five measurement locations were less than planned, typically ranging from 2.5 to 3.5 mm. The postimplant margin at the base was smaller than expected due to source placement errors, a correctable problem. CONCLUSIONS: Peripherally loaded Pd-103 prostate implants can deliver the prescription dose 3-5 mm outside the capsule, which is believed to be sufficient to treat 95-100% of EPE in favorable risk patients. However, dose coverage of EPE, like dose coverage of the prostate, is operator-dependent. PMID- 11728678 TI - Defining a dose-response relationship with radiotherapy for prostate cancer: is more really better? AB - PURPOSE: Data were reviewed addressing the association between radiation therapy (RT) dose and treatment outcome for localized prostate cancer to help clarify the existence of a potential dose-response relationship. METHODS AND MATERIALS: Articles were identified through the MEDLINE database, CancerLit database, and reference lists of relevant articles. Studies were categorized into four groups based upon the endpoint analyzed, including biochemical control (BC), local control (LC), pathologic control (PC), and cause-specific survival (CSS). The impact of increasing RT dose with each endpoint was recorded. RESULTS: Twenty-two trials involving a total of 11,297 patients were identified. Of the 11 trials addressing the association of RT dose with LC, 9 showed statistically significant improvements. Of the 12 trials that reported BC with RT dose, all showed statistically significant improvements. Two out of 4 studies analyzing PC with increasing dose showed a positive correlation. Finally, 3 out of 9 studies addressing RT dose with CSS showed statistically significant improvements. Despite inconclusive results, patients with poor risk features (e.g., prostate specific antigen [PSA] > or = 10, Gleason score [GS] > or = 7, or tumor stage > or = T2b) were most likely to benefit from increasing dose with respect to each endpoint. However, the optimal RT dose and the magnitude of benefit of dose escalation could not be identified. CONCLUSIONS: Although RT dose appears to correlate with various measures of treatment outcome, objective, high-quality data addressing this critical issue are still lacking. At the present time, the absolute improvement in outcome due to dose escalation, the subset of patients benefitting most, and the optimal dose remain to be defined. PMID- 11728679 TI - Response-adapted radiotherapy in the treatment of pediatric Hodgkin's disease: an interim report at 5 years of the German GPOH-HD 95 trial. AB - PURPOSE: A multinational trial on pediatric Hodgkin's disease (HD) with the aim to reduce the risk of long-term toxicity of combined modality treatment by restricting dose and volume of radiation therapy (RT) while maintaining the excellent treatment results of previous German multicenter trials (DAL-HD82-90). METHODS AND MATERIALS: Patients were treated according to stage of disease (CS) and defined risk factors in three treatment groups (TG) with 2, 4, or 6 cycles of combination chemotherapy. When a complete remission (CR) had been achieved, treatment was terminated without RT independent of initial stage or tumor bulk. Patients with a partial remission (PR) of >75% tumor regression were irradiated with 20 Gy using modified involved fields; in the case of PR <75% RT dose was 30 Gy, residual masses >50 mL received 35 Gy. RESULTS: From August 1995 to July 2000 a total of 956 patients have been registered, 830 as trial patients, 39% in TG1, 27% in TG2, 34% in TG3. 827 patients were evaluable by June 2001 with a median follow-up of 38 months. Chemotherapy (CTx) resulted in CR in 22%, PR >75% in 62%, PR <75% in 12%. Event-free survival (EFS) for the entire group is 90% (SD 0.01), for TG1 94%, TG2 91%, and TG3 84%; the overall survival is 97% in Kaplan-Meier analysis. Relapse-free survival (RFS) is superior for patients with RT after PR (93%) than for those without RT after CR (89%); the difference is significant (p = 0.01) for advanced stages, however not in TG1. Seventy-two events were observed by June 2001: 28 progressions during the initial therapy or within the first 3 months, 38 relapses, 3 second malignancies, three fatal accidents or infections; 18 patients have died. CONCLUSION: Treatment results of the GPOH-HD 95 trial are excellent thus far. The reduction of RT dose and volume in PR has not caused a significant impairment of overall and event-free survival in comparison to the previous German trials; however, failure rates are higher in advanced stages when RT is omitted after achieving a CR. It is too early to tell whether the HD 95 protocol will be successful in reducing late toxicity. PMID- 11728680 TI - Long-term results with radiotherapy for Stage I-II follicular lymphomas. AB - PURPOSE: To analyze the long-term results with radiotherapy (RT) for early-stage, low-grade follicular lymphomas. METHODS AND MATERIALS: From 1960 to 1988, 80 patients with Stage I (n = 33) or II (n = 47), World Health Organization Grade 1 (n = 50) or 2 (n = 30) follicular lymphoma were treated with RT. The lymph nodes or spleen were involved in 97% of cases. The maximal tumor sizes ranged from 0.5 to 11.0 cm (median 2.0). The RT fields encompassed only the involved Ann Arbor nodal region (involved-field RT) in 9% of the patients. The fields also included 1-3 adjacent, grossly uninvolved nodal regions (regional RT) in 54% of patients but were smaller than mantle or whole abdominopelvic fields. Mantle or whole abdominopelvic fields encompassing up to 6 grossly uninvolved regions (extended field RT) were used in the remaining 37% of patients. The total RT doses ranged from 26.2 to 50.0 Gy given in daily 1.0-3.0-Gy fractions. RESULTS: The follow-up of the surviving patients ranged from 3.5 to 28.7 years (median 19.0). No recurrences were found >17.0 years after RT, with 13 patients free of disease at their last follow-up visit 17.6-25.0 years after treatment. In 58% of cases, death was not from follicular lymphoma. The 15-year local control rate was 100% for 44 lymphomas <3.0 cm treated with only 27.8-30.8 Gy (median 30.0 in 20 fractions). Progression-free survival was affected by the maximal tumor size at the start of RT (15-year rate 49% vs. 29% for lymphomas <3.0 cm vs. > or =3.0 cm, respectively, p = 0.04) and Ann Arbor stage (15-year rate 66% vs. 26% for Stages I and II, respectively, p = 0.006). Ann Arbor stage also affected the cause specific survival (15-year rate 87% vs. 54% for Stages I and II, respectively, p = 0.01). No significant difference was found in overall survival between those treated with extended-field RT and those treated with involved-field RT or regional RT (15-year rate 49% and 40%, respectively, p = 0.51). The 15-year incidence rate of Grade 3 or greater late complications according to the Subjective, Objective, Management, and Analytical scale in patients treated with 26.2-30.8 Gy vs. 30.9-50.0 Gy was 0% and 6%, respectively. CONCLUSIONS: RT can cure approximately one half of Stage I and one quarter of Stage II, World Health Organization Grade 1 or 2 follicular lymphomas. Follicular lymphomas <3.0 cm can be controlled locally with doses of 27.8-30.8 Gy, and there is a trend toward a higher incidence of late complications with doses of >30.8 Gy. Doses of 25-30 Gy delivered in 15-20 fractions should be examined prospectively in patients with follicular lymphomas of <3.0 cm. PMID- 11728681 TI - Localized aggressive non-Hodgkin's lymphoma of the nasal cavity: a survey by the Japan Lymphoma Radiation Therapy Group. AB - PURPOSE: To clarify the role of radiotherapy and chemotherapy in the treatment of patients with localized aggressive non-Hodgkin's lymphomas (NHL) originating in the nasal cavity. METHODS AND MATERIALS: The survey, administered at 25 Japanese institutes in 1998, enabled us to collect the clinical data for 787 patients with localized aggressive NHL who were treated between 1988 and 1992. RESULTS: There were 42 patients (5%) with nasal lymphomas. Twelve of these patients received radiotherapy alone, and 30 were treated with a combination of radiotherapy and chemotherapy. The median radiation dose was 47 Gy (22-66). Twelve patients were reviewed histopathologically according to REAL (Revised European-American Classification of Lymphoid neoplasms) classification. T-cell or natural killer (NK) cell lymphomas were detected in 9 patients (75%), and diffuse large B-cell lymphomas in 3 (25%). The 5-year overall and disease-free survival (DFS) rates of all patients were 57% and 59%, respectively. The 5-year DFS rate for the 30 patients treated with the combined therapy was 64% and that for the 12 patients treated with radiotherapy alone was 46% (p = 0.021). For the 34 patients with stage-modified International Prognostic Index (m-IPI) 0-1, the 5-year DFS rates of those treated with the combined therapy and radiotherapy alone were 68% and 45%, respectively (p = 0.020), but there was no difference in DFS rate among the two groups of patients with m-IPI 2-3. The 5-year local control rates of the patients who received >46 Gy (n = 22) and < or =46 Gy (n = 20) were 95% and 76% (p = 0.087), respectively. There was no significant difference among the 5-year DFS rates (62% vs. 67%) and local control rates (87% vs. 100%) of the patients with T-cell or NK-cell lymphoma and diffuse large B-cell lymphoma. CONCLUSIONS: Patients with nasal lymphomas (m-IPI 0-1) should be treated with combined therapy. For the patients with high risk (m-IPI 2-3), the effectiveness of combined therapy was not clarified because of the small number of the patients. A high radiation dose >46 Gy may need to be used to achieve good local control. PMID- 11728682 TI - Combined p21WAF1/CIP1 and p53 overexpression predict improved survival in muscle invasive bladder cancer treated by radical radiotherapy. AB - PURPOSE: The prognostic value of p21 and p53 expression was evaluated for patients with muscle-invasive bladder cancer treated by radical radiotherapy. METHODS AND MATERIALS: Sixty-eight paraffin-embedded sections from surgically resected tumors taken prior to irradiation were immunostained for p21 and p53. RESULTS: Nuclear staining for p21 and p53 was demonstrated in 32/68 (47%) and 46/68 (68%) tumors, respectively. There was no correlation between p21 and p53 immunopositivity in this group (r = 0.067, p = 0.56). Patients were stratified into four distinct groups depending on staining for p21 and p53: p21+p53+, p21+p53-, p21-p53+, and p21-p53-. Patients with p21+p53+ tumors had the best prognosis with a 3-year survival of 82% compared to 12% for p21-p53+ tumors (p = 0.0031), 29% for p21+p53- tumors (p = 0.0108); and 45% for p21-p53- tumors (p = 0.0375). The p21+p53+ group also demonstrated significantly improved survival when a combined analysis was performed of p21-p53+, p21-p53-, and p21+p53- tumors (3-year survival = 30%, p = 0.0062). In a multivariate model, p21+p53+ tumors (p = 0.0108, relative risk [RR] = 5.18) and complete/partial response (p = 0.0019, RR = 3.76) were the only independent predictors of improved survival. CONCLUSIONS: With muscle-invasive bladder tumors treated by radical radiotherapy, stratification for p21 and p53 identifies distinct prognostic groups, with p21+p53+ tumors being associated with the best survival and p21-p53+ the worst. PMID- 11728683 TI - Long-term urinary function after transperineal brachytherapy for patients with large prostate glands. AB - PURPOSE: To summarize longer-term postbrachytherapy morbidity in patients with prostate glands >50 cm3. METHODS AND MATERIALS: From 1997 to 1998, 33 patients with a transrectal ultrasound-based prostate volume >50 cm3 were treated at the University of Washington by 125I (144 Gy) or 103Pd (115 Gy) implantation for prostate carcinoma. These 33 patients comprised 7% of the total implant patient population. Twelve patients were treated with neoadjuvant androgen ablation before implantation. The (125)I source strength ranged from 0.34 to 0.5 mCi and the 103Pd source strength ranged from 1.1 to 1.4 mCi (pre-NIST-99). The total number of sources implanted was 94-223 (median 155). Despite the typical implant related volume increase, the postimplant CT-defined prostate volumes were generally well-covered by the prescription isodose (median coverage 92%, range 80 100%). The preimplant urinary obstructive symptoms were quantified by the criteria of the American Urological Association. RESULTS: Of the 33 patients, 12 developed acute postimplant urinary retention, all presenting within 24 h of implantation. Patients who developed postimplant retention lasting >1 week were generally treated with intermittent self-catheterization. By 1 month, 85% of patients were catheter free. By 1 year, only 1 patient (4%) remained in urinary retention; the remainder of cases had resolved spontaneously. With follow-up of 1.7-2.6 years, the last American Urological Association scores were higher than the pretreatment scores in 15 patients and lower in 7 patients. No patient developed permanent urinary incontinence. Long-term changes in the American Urological Association scores were unrelated to whether the patient had been in urinary retention after implantation. Two patients developed rectal fistulas; they had preimplant transrectal ultrasound prostate volumes of 53 and 59 cm3, in the low range for this group of patients. No other patient had persistent rectal bleeding suggestive of clinically significant proctitis. The pretreatment serum prostate-specific antigen level was 3.3-15 ng/mL (median 7.2) and the last serum prostate-specific antigen level 0.1-1.6 ng/mL (median 0.2). CONCLUSION: Patients with larger prostate volumes appear to have moderate morbidity and a satisfactory technical outcome with brachytherapy. We do not believe the occurrence of two severe rectal complications was related to the prostate volume per se. Our experience and that of others calls into question the validity of using prostate volume as a criterion for patient suitability for prostate brachytherapy. PMID- 11728684 TI - The morbidity of treatment for patients with Stage I endometrial cancer: results from a randomized trial. AB - PURPOSE: To compare the treatment complications for patients with Stage I endometrial cancer treated with surgery and pelvic radiotherapy (RT) or surgery alone in a multicenter randomized trial. METHODS AND MATERIALS: The Postoperative Radiation Therapy in Endometrial Carcinoma (PORTEC) trial included patients with endometrial cancer confined to the uterine corpus, either Grade 1 or 2 with more than 50% myometrial invasion, or Grade 2 or 3 with less than 50% myometrial invasion. Surgery consisted of an abdominal hysterectomy and oophorectomy, without lymphadenectomy. After surgery, patients were randomized to receive pelvic RT (46 Gy), or no further treatment. A total of 715 patients were randomized. Treatment complications were graded using the French-Italian glossary. RESULTS: The analysis was done at a median follow-up duration of 60 months. 691 patients were evaluable. Five-year actuarial rates of late complications (Grades 1-4) were 26% in the RT group and 4% in the control group (p < 0.0001). Most were Grade 1 complications, with 5-year rates of 17% in the RT group and 4% in the control group. All severe (Grade 3-4) complications were observed in the RT group (3%). Most complications were of the gastrointestinal tract. The symptoms resolved after some years in 50% of the patients. Grade 1-2 genitourinary complications occurred in 8% of the RT patients, and 4% of the controls. Bone complications occurred in 4 RT patients (1%). Seven patients (2%) discontinued their RT due to acute RT-related symptoms. Patients with acute morbidity had an increased risk of late RT complications (p = 0.001). The 4-field box technique was associated with a lower risk of late complications (p = 0.06). CONCLUSION: Pelvic RT increases the morbidity of treatment in Stage I endometrial cancer. In the PORTEC trial, severe complications occurred in 3% of treated patients, and over 20% experienced mild (mostly Grade 1) symptoms. Patients with acute RT-related morbidity had an increased risk of late complications. As pelvic RT in Stage I endometrial carcinoma was shown to significantly reduce the rate of locoregional recurrence, but without a survival benefit, its use in the adjuvant setting requires careful patient selection (treating those at increased risk of relapse), and the use of treatment schemes with the lowest risk of morbidity. PMID- 11728685 TI - Effective palliative radiation therapy in advanced and recurrent ovarian carcinoma. AB - PURPOSE: To retrospectively review our experience using radiation therapy as a palliative treatment in ovarian carcinoma. METHODS AND MATERIALS: Eighty patients who received radiation therapy for ovarian carcinoma between 1983 and 1998 were reviewed. The indications for radiation therapy, radiation therapy techniques, details, tolerance, and response were recorded. A complete response required complete resolution of the patient's symptoms, radiographic findings, palpable mass, or CA-125 level. A partial response required at least 50% resolution of these parameters. The actuarial survival rates from initial diagnosis and from the completion of radiation therapy were calculated. RESULTS: The median age of the patients was 67 years (range 26 to 90 years). A median of one laparotomy was performed before irradiation. Zero to 20 cycles of a platinum-based chemotherapy regimen were delivered before irradiation (median = 6 cycles). The reasons for palliative treatment were: pain (n = 22), mass (n = 23), obstruction of ureter, rectum, esophagus, or stomach (n = 12), a positive second-look laparotomy (n = 9), ascites (n = 8), vaginal bleeding (n = 6), rectal bleeding (n = 1), lymphedema (n = 3), skin involvement (n = 1), or brain metastases with symptoms (n = 11). Some patients received treatment for more than one indication. Treatment was directed to the abdomen or pelvis in 64 patients, to the brain in 11, and to other sites in 5. The overall response rate was 73%. Twenty-eight percent of the patients experienced a complete response of their symptoms, palpable mass, and/or CA-125 level. Forty-five percent had a partial response. Only 11% suffered progressive disease during therapy that required discontinuation of the treatment. Sixteen percent had stable disease. The duration of the responses and stable disease lasted until death except in 10 patients who experienced recurrence of their symptoms between 1 and 21 months (median = 9 months). The 1-, 2-, 3-, and 5-year actuarial survival rates from diagnosis were 89%, 73%, 42%, and 33%, respectively. The survival rates calculated from the completion of radiotherapy were 39%, 27%, 13%, and 10%, respectively. Five percent of patients experienced Grade 3 diarrhea, vomiting, myelosuppression, or fatigue. Fourteen percent of patients experienced Grade 1 or 2 diarrhea, 19% experienced Grade 1 or 2 nausea and vomiting, and 11% had Grade 1 or 2 myelosuppression. CONCLUSIONS: In this series of radiation therapy for advanced ovarian carcinoma, the response, survival, and tolerance rates compare favorably to those reported for current second- and third-line chemotherapy regimens. Cooperative groups should consider evaluating prospectively the use of radiation therapy before nonplatinum and/or nonpaclitaxel chemotherapy in these patients. PMID- 11728686 TI - Pathologic downstaging of T3-4Nx rectal cancer after chemoradiation: 5 fluorouracil vs. Tegafur. AB - PURPOSE: To describe downstaging effects in locally advanced rectal cancer induced by 2 fluopirimidine radiosensitizing agents given through different routes in conjunction with preoperative radiotherapy. METHODS AND MATERIALS: From March 1995 to December 1999, two consecutive groups of patients with cT3-4Nx rectal cancer (94% CT scan, 71% endorectal ultrasound) were treated with either (1) 45-50 Gy (1.8 Gy/day, 25 fractions) and 5-fluorouracil (5-FU) (500-1,000 mg/m2 by 24-h continuous i.v. infusion on Days 1-4 and 21-25) or (2) oral Tegafur (1,200 mg/day on Days 1-35, including weekends). Surgery was performed 4 to 6 weeks after the completion of chemoradiation. RESULTS: The total T downstaging rate was 46% in the 5-FU group and 53% in the Tegafur group. Subcategories were downstaged by the sensitizing agents (5-FU vs. Tegafur) as follows: pT0-1, 14% vs. 23%; pT2, 32% vs. 32%; pT3, 49% vs. 37%; pT4, 5% vs. 7%; and N(0), 74% vs. 86%. Analysis of residual malignant disease in the specimen discriminated mic/mac subgroups (mic: <20% of microscopic cancer residue), with evident superior downstaging effects in the Tegafur-treated group: pTmic 23% vs. 58% (p = 0.002). CONCLUSIONS: When administered concurrent with pelvic irradiation, oral Tegafur induced downstaging rates in both T and N categories superior to those induced by intermediate doses of 5-FU by continuous i.v. infusion. In this pilot experience, oral Tegafur reproduced the characteristics of downstaging described previously when full doses of 5-FU have been combined with radiotherapy. PMID- 11728687 TI - The position and volume of the small bowel during adjuvant radiation therapy for rectal cancer. AB - PURPOSE: The rate of small bowel toxicity from adjuvant pelvic radiation therapy (RT) for rectal cancer has been reported to be lower for patients treated preoperatively (Preop). This was probably due to a lesser volume of irradiated small bowel; however, studies of postoperative treatment reported that patients with an abdominoperineal resection (APR), who likely have the largest volume of small bowel in the pelvis, had less acute and chronic toxicity than those with a low anterior resection (LAR). In this study, three-dimensional treatment planning techniques were used to characterize the position and volume of small bowel in the pelvis and compare these to repeat studies obtained during the typical 5-week course of treatment to attempt to explain the above observations. METHODS AND MATERIALS: Treatment planning CT scans were obtained in 30 patients with rectal cancer (10 Preop, 10 LAR, 10 APR), including 12 patients with weekly CT scans during RT (65 scans). The position of the small bowel was measured by the distance to the nearest small bowel from the bones of the posterior pelvis and by the volume of small bowel within four anatomically defined regions of the pelvis. The motion of the small bowel was expressed as the standard deviation of the small bowel position measured with both the distance and the volume in the 12 patients with repeat studies. RESULTS: Contrast-containing small bowel was found an average 2.9 cm more anterior than small bowel without contrast below the sacral promontory. The position of the small bowel in Preop patients was significantly more anterior (p < or = 0.01) with less volume (p < or = 0.04) in the pelvis than postoperatively treated patients. The small bowel was also more anterior for patients with an LAR vs. APR (p < or = 0.03) but with similar volume in all pelvic regions. Small bowel motion, expressed as the standard deviation of the distance from the bones of the posterior pelvis to the closest small bowel, was 2.9 cm, 1.4 cm, and 0.2 cm for the Preop, LAR, and APR group, respectively. The LAR group had a considerable degree of motion in the posterior pelvis. Increased bladder volume was associated with reduced small bowel volumes, although this benefit decreased during treatment. CONCLUSION: Because treatment planning CT scans can detect small bowel that does not contain contrast, they may be more accurate than the traditional small bowel series. The Preop patients had significantly less pelvic small bowel supporting the clinical observation of better tolerance to therapy. The higher small bowel toxicity reported for LAR vs. APR patients may be explained by the greater variability of both the position and volume of the small bowel in the posterior pelvis for LAR patients. This finding suggests that a single planning study may not be accurate for the block design used for boost treatment of LAR patients. Bladder-filling techniques were useful for Preop and LAR but not APR patients, and decreased in benefit over time. This study suggested that treatment planning CT scans were more useful than a small bowel series and that more than one treatment planning CT may be obtained in any patient receiving > 45 Gy for rectal cancer. However, further research will be necessary to determine the optimal timing and total number of repeat studies. PMID- 11728688 TI - Phase I study of twice-weekly gemcitabine and concurrent thoracic radiation for patients with locally advanced non-small-cell lung cancer. AB - PURPOSE: To determine the maximum tolerated dose (MTD) and dose-limiting toxicity of twice-weekly gemcitabine and concurrent thoracic radiation in patients with Stage IIIa/IIIb non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: Seventeen patients with histologically confirmed Stage IIIa and IIIb NSCLC were studied. Gemcitabine was administered via a 30-min i.v. infusion twice weekly for 6 weeks concurrent with 60 Gy of thoracic radiation. Gemcitabine, starting at a twice-weekly dose of 10 mg/m2 (20 mg/m2/week), was escalated in 10-15 mg/m2 increments in successive cohorts of 3 to 6 patients until dose-limiting toxicity was observed. RESULTS: Of the 17 patients entered, 16 were evaluable for toxicity. The dose-limiting toxicity at 50 mg/m2 given twice weekly (100 mg/m2/week) was Grade 3 pneumonitis observed in 1 patient, Grade 3 pulmonary fibrosis in a second patient, and Grade 4 esophagitis observed in two additional patients. Twice-weekly gemcitabine at a dose of 35 mg/m2 was determined to be the MTD. The overall response rate for the 16 evaluable patients was 88%. The median survival for the entire group is 16.0 months. CONCLUSIONS: The MTD of twice weekly gemcitabine is 35 mg/m2 (70 mg/m2/week) given with thoracic radiation. A Phase II study within the Cancer and Leukemia Group B to ascertain the potential efficacy of this treatment regimen is in development. PMID- 11728689 TI - Field size reduction enables iso-NTCP escalation of tumor control probability for irradiation of lung tumors. AB - PURPOSE: With the mean lung dose (MLD) as an estimator for the normal tissue complication probability (NTCP) of the lung, we assessed whether the probability of tumor control of lung tumors might be increased by dose escalation in combination with a reduction of field sizes, thus increasing target dose inhomogeneity while maintaining a constant MLD. METHODS AND MATERIALS: An 8-MV AP PA irradiation of a lung tumor, located in a cylindrically symmetric lung equivalent phantom, was modeled using numerical simulation. Movement of the clinical target volume (CTV) due to patient breathing and setup errors was simulated. The probability of tumor control, expressed as the equivalent uniform dose (EUD) of the CTV, was assessed as a function of field size, under the constraint of a constant MLD. The approach was tested for a treatment of a non small cell lung cancer (NSCLC) patient using the beam directions of the clinically applied treatment plan. RESULTS: In the phantom simulation it was shown that by choosing field sizes that ensured a minimum dose of 95% in the CTV ("conventional" plan) taking into account setup errors and tumor motion, an EUD of the CTV of 43.8 Gy can be obtained for a prescribed dose of 44.2 Gy. By reducing the field size and thus shifting the 95% isodose surface inwards, the EUD increases to a maximum of 68.3 Gy with a minimum dose in the CTV of 55.2 Gy. This increase in EUD is caused by the fact that field size reduction enables escalation of the prescribed dose while maintaining a constant MLD. Further reduction of the field size results in decrease of the EUD because the minimum dose in the CTV becomes so low that it has a predominant effect on the EUD, despite further escalation of the prescribed dose. For the NSCLC patient, the EUD could be increased from an initial 62.2 Gy for the conventional plan, to 83.2 Gy at maximum. In this maximum, the prescribed dose is 88.1 Gy, and the minimum dose in the CTV is 67.4 Gy. In this case, the 95% isodose surface is conformed closely to the "static" CTV during treatment planning. CONCLUSIONS: Iso-NTCP escalation of the probability of tumor control is possible for lung tumors by reducing field sizes and allowing a larger dose inhomogeneity in the CTV. Optimum field sizes can be derived, having the highest EUD and highest minimum dose in the CTV under condition of a constant NTCP of the lungs. We conclude that the concept of homogeneous dose in the target volume is not the best approach to reach the highest probability of tumor control for lung tumors. PMID- 11728690 TI - RTOG 96-10: reirradiation with concurrent hydroxyurea and 5-fluorouracil in patients with squamous cell cancer of the head and neck. AB - PURPOSE: Patients with recurrent squamous cell cancer of the head and neck (SCH&N) are generally treated with systemic chemotherapy. Improvement in survival has not occurred, despite an increased objective response rate. This study was undertaken to explore the feasibility and toxicity, and estimate the therapeutic impact of, reirradiation (RRT) with concurrent hydroxyurea and 5-fluorouracil. METHODS AND MATERIALS: The eligibility requirements included SCH&N presenting as a second primary or recurrence > or =6 months after definitive RT to > or =45 Gy, with > or =75% of the tumor volume within the previous field. The cumulative spinal cord dose was limited to 50 Gy, and measurable disease was required. Four weekly cycles were given, each separated by 1 week of rest. A cycle consisted of 5 days, Monday through Friday, of 1.5-Gy twice-daily repeated RT, with the fractions separated by > or =6 h, with 1.5 g of hydroxyurea given 2 h and 300 mg/m2 of a 5-fluorouracil IV bolus given 30 min before each second daily fraction. RESULTS: Eighty-six patients were entered; 81 patients were assessable. The median prior radiation dose was 61.2 Gy. The 4 planned cycles were delivered in 79% of patients. Grade 3 mucositis occurred in 14% of patients, and Grade 4 in 5%. Grade 3 acute pharyngeal toxicity was seen in 17%. Grade 3 neutropenia occurred in 9%, Grade 4 in 10%, and Grade 5 in 7%. Six patients died of treatment related toxicity. Two died of hemorrhage from the tumor site without thrombocytopenia. With a median follow-up of 16.3 months for living patients, the estimated median overall survival was 8.2 months and the estimated 1-year survival rate 41.7%. Patients treated >3 years after the previous RT had a 1-year survival rate of 48% compared with 35% for patients treated within 3 years (p = 0.017). The 1-year survival rate for patients with a second primary was 54% compared with 38% for patients with recurrence (p = 0.083). CONCLUSION: Repeated RT with concurrent chemotherapy as given in this study is a feasible approach for selected, previously irradiated patients with SCH&N and may produce increased median and 1-year survival rates compared with systemic chemotherapy trials reported in the literature. A randomized study should be conducted to compare these two different approaches. PMID- 11728691 TI - Cervical lymph node relapses of head-and-neck squamous cell carcinoma: is brachytherapy a therapeutic option? AB - PURPOSE: To report the results of interstitial brachytherapy (IBT) without salvage surgery for isolated cervical lymph node relapses. PATIENT AND METHODS: From 1970 to 1989, 84 patients were treated; 76 patients had relapsed in sites of previous external beam radiation. In 72 patients, IBT was sole salvage treatment (mean, 56.5 Gy). In 12 patients IBT (mean, 38 Gy) was combined with further external beam radiotherapy (mean, 41 Gy). RESULTS: Local control in the neck was 49% at 1 year, 31% at 2 years, and 0% at 5 years. Overall survival was 33% at 1 year, 13% at 2 years, and 1% at 5 years. Significant toxicity occurred in 35% (7% fatal). Multivariate analysis shows survival after salvage was better for patients who had achieved initial control for > or =18 months before relapse (0% vs. 13% at 3 years, p < 0.0002). Lymph node control was better for patients who received total salvage dose > or =60 Gy (0% vs. 56% at 3 years, p = 0.0004). CONCLUSION: Given its poor efficiency and its toxicity, IBT must be considered only when surgery is contraindicated and if lymph node relapse occurs after a minimal interval of 18 months. PMID- 11728692 TI - Dose conformity of gamma knife radiosurgery and risk factors for complications. AB - PURPOSE: To quantitatively evaluate dose conformity achieved using Gamma Knife radiosurgery, compare results with those reported in the literature, and evaluate risk factors for complications. METHODS AND MATERIALS: All lesions treated at our institution with Gamma Knife radiosurgery from May 1993 (when volume criteria were routinely recorded) through December 1998 were reviewed. Lesions were excluded from analysis for reasons listed below. Conformity index (the ratio of prescription volume to target volume) was calculated for all evaluable lesions and for lesions comparable to those reported in the literature on conformity of linac radiosurgery. Univariate Cox regression models were used to test for associations between treatment parameters and toxicity. RESULTS: Of 1612 targets treated in 874 patients, 274 were excluded, most commonly for unavailability of individual prescription volume data because two or more lesions were included within the same dose matrix (176 lesions), intentional partial coverage for staged treatment of large arteriovenous malformations (AVMs) (33 lesions), and missing target volume data (26 lesions). The median conformity indices were 1.67 for all 1338 evaluable lesions and 1.40-1.43 for lesions comparable to two linac radiosurgery series that reported conformity indices of 1.8 and 2.7, respectively. Among all 651 patients evaluable for complications, there were one Grade 5, eight Grade 4, and 27 Grade 3 complications. Increased risk of toxicity was associated with larger target volume, maximum lesion diameter, prescription volume, or volume of nontarget tissue within the prescription volume. CONCLUSIONS: Gamma Knife radiosurgery achieves much more conformal dose distributions than those reported for conventional linac radiosurgery and somewhat more conformal dose distributions than sophisticated linac radiosurgery techniques. Larger target, nontarget, or prescription volumes are associated with increased risk of toxicity. PMID- 11728693 TI - Radiosurgery for brain metastases: who may not benefit? AB - PURPOSE: To select a group of patients with brain metastases for whom stereotactic radiosurgery (SRS) may not be beneficial. PATIENTS, MATERIALS, AND METHODS: Actuarial survival of 87 patients with brain metastases treated with SRS between July 1993 and May 1999 was retrospectively analyzed under stratification by the Score Index for Stereotactic Radiosurgery for Brain Metastases (SIR). To identify the group of patients most likely to survive less than 6 months after SRS, Cox model survival curves were calculated for all SIR values, and Kaplan Meier survival curves were calculated for two SIR subsets (0-5 and 6-10) and were compared by log-rank test. RESULTS: Overall median survival after SRS was 6.88 months. The stratification of patients into two SIR subsets (0-5 and 6-10) sustained statistical significance regarding survival with p = 0.0001. The median survival time for the group of patients with SIR between 0 and 5 was 4.52 months (95% confidence interval of 2.82 to 5.84 months). Survival probability at 6 months for this group of patients with poor prognosis was 35.6%. CONCLUSION: Patients with brain metastases and SIR of 5 or lower have an expected median survival of less than 6 months after treatment with radiosurgery. Thus, radiosurgery may not be beneficial for this group of patients. PMID- 11728694 TI - Influence of irradiation on therapy-associated psychological distress in breast carcinoma patients. AB - PURPOSE: To confirm our assumptions regarding factors that apparently cause psychological distress related to adjuvant radiotherapy in breast cancer patients and to evaluate variables that can predict therapy-associated distress. METHODS AND MATERIALS: Between January 1997 and April 1998, 111 women (33-84 years) with early-stage breast cancer were irradiated (56 Gy) after breast-conserving surgery. Patients were given self-assessment questionnaires on the first and last day of radiotherapy. Statistical analysis was performed using the structural equation model LISREL, variance analysis, and regression analysis. RESULTS: The internal subject-related factors (coping, radiation-related anxiety, physical distress, psychological distress) reciprocally influenced each other, whereas external radiotherapy-specific factors (environmental influence, confidence in the medical staff) were causally related to coping, anxiety, and distress. Fifty three percent of the women felt distressed because cancer affected the breast; 48% were initially afraid of radiotherapy. For 36%, anxiety was not reduced during treatment. Highly distressed women were identified by the following parameters: < or =58 years; initial anxiety; they were affected by having breast cancer, were negatively affected by environmental factors, and did not find distraction helpful. CONCLUSION: Despite considerable individual variability in breast cancer patients, it seems possible to identify women who run a high risk of therapy-associated distress. In these patients, psychosocial support is necessary to reduce treatment-related anxiety and to stabilize confidence in the medical staff. PMID- 11728695 TI - Additional gene therapy with Ad5CMV-p53 enhanced the efficacy of radiotherapy in human prostate cancer cells. AB - PURPOSE: The aim of this study was to investigate the efficacy of combination therapy of ionizing radiation (IR) and adenoviral p53 gene therapy and to evaluate its molecular mechanisms. METHODS AND MATERIALS: Two human prostate cancer cell lines, DU145 and PC-3 cells, containing different types of p53 gene mutations, were investigated. The recombinant adenovirus vector containing the wild-type p53 gene (Ad5CMV-p53) was used for this study. Cells were irradiated (in 0, 2, 4, and 6 Gy, 300 cGy/min) and after 12 h of irradiation, the cells were infected with various doses of Ad5CMV-p53 (0-40 multiplicity of infection [MOI]). Cytotoxicity was determined by clonogenic assay. The molecular mechanisms were evaluated by quantitative reverse transcriptase-polymerase chain reaction (RT PCR), apoptotic cell detection, and cell cycle analysis. RESULTS: The cell growth inhibition in DU145 (p53-mutated) cells by IR was strongly enhanced by additional Ad5CMV-p53 infection in a viral dose-dependent manner. In DU145 cells, IR alone induced minimal p53 mRNA expression. However, IR combined with Ad5CMV-p53 infection stimulated significant increase in p53 mRNA expression supplemented with Bax and p21 mRNA expressions. In PC-3 (p53-null), IR induced Bax and p21 mRNA expression, while the combination effects were observed in p53, Bax, and p21 mRNA expression. Apoptotic cell deaths were rarely observed after IR alone (DU145: 3%, PC-3: 5%). However, after combination therapy, the proportion of apoptotic cells greatly increased (sevenfold in DU145 cells, and twice in PC-3 cells). G1 cell cycle arrest was observed after Ad5CMV-p53 infection and the combination in both cell lines. CONCLUSION: In this study, we demonstrated that the combination of IR and Ad5CMV-p53 gene therapy resulted in remarkable synergistic effects in human prostate cancer cells. This combination therapy could be one of the optimal treatment strategies for radioresistant prostate cancer. PMID- 11728696 TI - Patterns of proliferation related to vasculature in human head-and-neck carcinomas before and after transplantation in nude mice. AB - PURPOSE: The predictive potential of tumor cell kinetic parameters may be improved when they are studied in relation to other microenvironmental parameters. The purpose of this investigation was to quantitatively categorize human tumor samples according to proliferation patterns. Second, it was examined whether these characteristics are retained after xenotransplantation. METHODS AND MATERIALS: Fifty tumor samples from head-and-neck cancer patients were immunohistochemically stained for Ki-67 and vessels. Also, parts of the samples were transplanted into nude mice. Tumors were categorized according to previously described patterns of proliferation. Vascular and proliferation patterns were analyzed using an image processing system. RESULTS: The 50 tumors were categorized into four patterns of proliferation by visual assessment: marginal (6), intermediate (10), random (21), and mixed (12). One tumor could not be classified. These patterns were quantified by calculating the Ki-67 labeling index in distinct zones at increasing distance from vessels yielding good discrimination and significant differences between patterns. The probability of growth after xenotransplantation was significantly higher for tumors with a labeling index and vascular density above the median value compared to tumors with both parameters below the median (82% vs. 35%). Fifty percent of the tumors retained their proliferation patterns after xenotransplantation. CONCLUSION: The categorization by proliferation pattern previously described by others was reproduced quantitatively and spatially related to the vascular network using a computerized image processing system. The combination of quantitative and architectural information of multiple microenvironmental parameters adds a new dimension to the study of treatment resistance mechanisms. Tumor models representative of the various patterns can be used to further investigate the relevance of these architectural patterns. PMID- 11728697 TI - Constitutive NF-kappaB activity influences basal apoptosis and radiosensitivity of head-and-neck carcinoma cell lines. AB - PURPOSE: Nuclear factor-kappaB (NF-kappaB) has been implicated in anti-apoptotic gene transactivation, according to its transcriptional activity. The present study was designed to investigate whether constitutive NF-kappaB activity could modulate basal apoptosis and intrinsic radiosensitivity of KB head-and-neck carcinoma cell line and KB3 subline. The KB3 subline was more radiosensitive (SF2 = 0.48, alpha = 0.064) than the radioresistant KB parental cell line (SF2 = 0.80, alpha = 0.114). METHODS AND MATERIALS: Constitutive NF-kappaB DNA-binding activity was determined using electrophoretic mobility shift assay. Modulation of NF-kappaB activity was performed by exposing both cell lines to tumor necrosis factor alpha or dexamethasone. Apoptotic cell population was analyzed using flow cytometry (annexin V/propidium iodide). Radiosensitivity was assessed from determination of the surviving fraction at 2 Gy (SF2), and alpha and beta parameters were determined using the linear-quadratic model. RESULTS: Constitutive NF-kappaB activity was found to be significantly lower in KB3 than in KB. KB cell line exposure to dexamethasone significantly decreased NF-kappaB DNA-binding activity and, consequently, enhanced baseline apoptosis and radiosensitivity (alpha values: 0.114 vs. 0.052). Conversely, exposure of KB3 cells to tumor necrosis factor alpha increased NF-kappaB DNA-binding activity and resulted in a significant decrease (50%) in rate of apoptosis and in radiosensitivity (SF2 values: 0.48 vs. 0.63). CONCLUSIONS: Modulation of NF kappaB DNA-binding activity influences baseline apoptosis and intrinsic radiosensitivity. PMID- 11728698 TI - Pseudo beam's-eye-view as applied to beam orientation selection in intensity modulated radiation therapy. AB - PURPOSE: To introduce the concept of pseudo beam's-eye-view (pBEV), to establish a framework for computer-assisted beam orientation selection in intensity modulated radiation therapy (IMRT), and to evaluate the utility of the proposed technique. METHODS AND MATERIALS: To facilitate the selection of beam orientations for IMRT treatment planning, a scoring of beam direction was introduced. The score function was based on the maximum target dose deliverable by the beam without exceeding the tolerance doses of the critical structures. For the score function calculation, the beam portal at given gantry and couch angles was divided into a grid of beamlets. Each beamlet crossing the target was assigned the maximum intensity that could be used without exceeding the dose tolerances of the organs at risk (OARs) and normal tissue. Thereafter, a score was assigned to the beam according to the target dose delivered. The beams for the treatment were selected among those with the highest scores. In a sense, this technique is similar to the beam's-eye-view approach used in conventional radiation therapy, except that the evaluation by a human is replaced by a score function, and beam modulation is taken into account. RESULTS: The pBEV technique was tested on two clinical cases: a paraspinal treatment and a nasopharyngeal cancer with both coplanar and noncoplanar beam configurations. The plans generated under the guidance of pBEV for the paraspinal treatment offered superior target dose uniformity and reduced OAR doses. For the nasopharyngeal cancer case, it was also found that the pBEV-selected coplanar and noncoplanar beams significantly improved the target coverage without compromising the sparing of the OARs. CONCLUSIONS: The pBEV technique developed in this work provides a comprehensive tool for beam orientation selection in IMRT. It is especially valuable for complicated cases, where the target is surrounded by several sensitive structures and where it is difficult to select a set of good beam orientations. The pBEV technique has considerable potential for simplifying the IMRT treatment planning process and for maximizing the technical capacity of IMRT. PMID- 11728699 TI - Leaf position optimization for step-and-shoot IMRT. AB - PURPOSE: To describe the theoretical basis, the algorithm, and implementation of a tool that optimizes segment shapes and weights for step-and-shoot intensity modulated radiation therapy delivered by multileaf collimators. METHODS AND MATERIALS: The tool, called SOWAT (Segment Outline and Weight Adapting Tool) is applied to a set of segments, segment weights, and corresponding dose distribution, computed by an external dose computation engine. SOWAT evaluates the effects of changing the position of each collimating leaf of each segment on an objective function, as follows. Changing a leaf position causes a change in the segment-specific dose matrix, which is calculated by a fast dose computation algorithm. A weighted sum of all segment-specific dose matrices provides the dose distribution and allows computation of the value of the objective function. Only leaf position changes that comply with the multileaf collimator constraints are evaluated. Leaf position changes that tend to decrease the value of the objective function are retained. After several possible positions have been evaluated for all collimating leaves of all segments, an external dose engine recomputes the dose distribution, based on the adapted leaf positions and weights. The plan is evaluated. If the plan is accepted, a segment sequencer is used to make the prescription files for the treatment machine. Otherwise, the user can restart SOWAT using the new set of segments, segment weights, and corresponding dose distribution. The implementation was illustrated using two example cases. The first example is a T1N0M0 supraglottic cancer case that was distributed as a multicenter planning exercise by investigators from Rotterdam, The Netherlands. The exercise involved a two-phase plan. Phase 1 involved the delivery of 46 Gy to a concave-shaped planning target volume (PTV) consisting of the primary tumor volume and the elective lymph nodal regions II-IV on both sides of the neck. Phase 2 involved a boost of 24 Gy to the primary tumor region only. SOWAT was applied to the Phase 1 plan. Parotid sparing was a planning goal. The second implementation example is an ethmoid sinus cancer case, planned with the intent of bilateral visus sparing. The median PTV prescription dose was 70 Gy with a maximum dose constraint to the optic pathway structures of 60 Gy. RESULTS: The initial set of segments, segment weights, and corresponding dose distribution were obtained, respectively, by an anatomy-based segmentation tool, a segment weight optimization tool, and a differential scatter-air ratio dose computation algorithm as external dose engine. For the supraglottic case, this resulted in a plan that proved to be comparable to the plans obtained at the other institutes by forward or inverse planning techniques. After using SOWAT, the minimum PTV dose and PTV dose homogeneity increased; the maximum dose to the spinal cord decreased from 38 Gy to 32 Gy. The left parotid mean dose decreased from 22 Gy to 19 Gy and the right parotid mean dose from 20 to 18 Gy. For the ethmoid sinus case, the target homogeneity increased by leaf position optimization, together with a better sparing of the optical tracts. CONCLUSIONS: By using SOWAT, the plans improved with respect to all plan evaluation end points. Compliance with the multileaf collimator constraints is guaranteed. The treatment delivery time remains almost unchanged, because no additional segments are created. PMID- 11728700 TI - Uncertainties in model-based outcome predictions for treatment planning. AB - PURPOSE: Model-based treatment-plan-specific outcome predictions (such as normal tissue complication probability [NTCP] or the relative reduction in salivary function) are typically presented without reference to underlying uncertainties. We provide a method to assess the reliability of treatment-plan-specific dose volume outcome model predictions. METHODS AND MATERIALS: A practical method is proposed for evaluating model prediction based on the original input data together with bootstrap-based estimates of parameter uncertainties. The general framework is applicable to continuous variable predictions (e.g., prediction of long-term salivary function) and dichotomous variable predictions (e.g., tumor control probability [TCP] or NTCP). Using bootstrap resampling, a histogram of the likelihood of alternative parameter values is generated. For a given patient and treatment plan we generate a histogram of alternative model results by computing the model predicted outcome for each parameter set in the bootstrap list. Residual uncertainty ("noise") is accounted for by adding a random component to the computed outcome values. The residual noise distribution is estimated from the original fit between model predictions and patient data. RESULTS: The method is demonstrated using a continuous-endpoint model to predict long-term salivary function for head-and-neck cancer patients. Histograms represent the probabilities for the level of posttreatment salivary function based on the input clinical data, the salivary function model, and the three dimensional dose distribution. For some patients there is significant uncertainty in the prediction of xerostomia, whereas for other patients the predictions are expected to be more reliable. In contrast, TCP and NTCP endpoints are dichotomous, and parameter uncertainties should be folded directly into the estimated probabilities, thereby improving the accuracy of the estimates. Using bootstrap parameter estimates, competing treatment plans can be ranked based on the probability that one plan is superior to another. Thus, reliability of plan ranking could also be assessed. CONCLUSIONS: A comprehensive framework for incorporating uncertainties into treatment-plan-specific outcome predictions is described. Uncertainty histograms for continuous variable endpoint models provide a straightforward method for visual review of the reliability of outcome predictions for each treatment plan. PMID- 11728701 TI - The effect of setup uncertainty on normal tissue sparing with IMRT for head-and neck cancer. AB - PURPOSE: Intensity-modulated radiotherapy (IMRT) is being evaluated in the management of head-and-neck cancers at several institutions, and a Radiation Therapy Oncology Group study of its utility in parotid sparing is under development. There is an inherent risk that the sharper dose gradients generated by IMRT amplify the potentially detrimental impact of setup uncertainty. The International Commission on Radiation Units and Measurements Report 62 (ICRU-62) defined planning organ-at-risk volume (PRV) to account for positional uncertainties for normal tissues. The purpose of this study is to quantify the dosimetric effect of employing PRV for the parotid gland and to evaluate the use of PRV on normal-tissue sparing in the setting of small clinical setup errors. METHODS AND MATERIALS: The optimized nine-beam IMRT plans for three head-and-neck cancer patients participating in an institutional review board approved parotid sparing protocol were used as reference plans. A second optimized plan was generated for each patient by adding a PRV of 5 mm for the contralateral parotid gland. The effect of these additions on the quality of the plans was quantified, in terms of both target coverage and normal-tissue sparing. To test the value of PRV in a worst-case scenario, systematic translational setup uncertainties were simulated by shifting the treatment isocenter 5 mm superiorly, inferiorly, left, right, anteriorly, and posteriorly, without altering optimized beam profiles. At each shifted isocenter, dose distributions were recalculated, producing a total of six shifted plans without PRV and six shifted plans with PRV for each patient. The effect of setup uncertainty on parotid sparing and the value of PRV in compensating for the uncertainty were evaluated. RESULTS: The addition of the PRV and reoptimization did not significantly affect the dose to gross tumor volume, spinal cord, or brainstem. In contrast, without any shift, the PRV did increase parotid sparing and reduce coverage of the nodal region adjacent to the parotid gland. As expected, when the plans were shifted, the greatest increase in contralateral parotid irradiation was noted with shifts toward the contralateral parotid gland. With these shifts, the average volume of contralateral parotid receiving greater than 30 Gy was reduced from 22% to 4% when a PRV was used. This correlated with a reduction in the average normal-tissue complication probability (NTCP) from 22% to 7%. CONCLUSIONS: The use of PRV may limit the volume of normal tissue structures, such as the parotid gland, exceeding tolerance dose as a result of setup errors. Consequently, it will be important to incorporate the nomenclature of ICRU-62 into the design of future IMRT studies, if the clinical gains of increased normal-tissue sparing are to be realized. PMID- 11728702 TI - The reproducibility of organ position using active breathing control (ABC) during liver radiotherapy. AB - PURPOSE: To evaluate the intrafraction and interfraction reproducibility of liver immobilization using active breathing control (ABC). METHODS AND MATERIALS: Patients with unresectable intrahepatic tumors who could comfortably hold their breath for at least 20 s were treated with focal liver radiation using ABC for liver immobilization. Fluoroscopy was used to measure any potential motion during ABC breath holds. Preceding each radiotherapy fraction, with the patient setup in the nominal treatment position using ABC, orthogonal radiographs were taken using room-mounted diagnostic X-ray tubes and a digital imager. The radiographs were compared to reference images using a 2D alignment tool. The treatment table was moved to produce acceptable setup, and repeat orthogonal verification images were obtained. The positions of the diaphragm and the liver (assessed by localization of implanted radiopaque intra-arterial microcoils) relative to the skeleton were subsequently analyzed. The intrafraction reproducibility (from repeat radiographs obtained within the time period of one fraction before treatment) and interfraction reproducibility (from comparisons of the first radiograph for each treatment with a reference radiograph) of the diaphragm and the hepatic microcoil positions relative to the skeleton with repeat breath holds using ABC were then measured. Caudal-cranial (CC), anterior-posterior (AP), and medial-lateral (ML) reproducibility of the hepatic microcoils relative to the skeleton were also determined from three-dimensional alignment of repeat CT scans obtained in the treatment position. RESULTS: A total of 262 fractions of radiation were delivered using ABC breath holds in 8 patients. No motion of the diaphragm or hepatic microcoils was observed on fluoroscopy during ABC breath holds. From analyses of 158 sets of positioning radiographs, the average intrafraction CC reproducibility (sigma) of the diaphragm and hepatic microcoil position relative to the skeleton using ABC repeat breath holds was 2.5 mm (range 1.8-3.7 mm) and 2.3 mm (range 1.2 3.7 mm) respectively. However, based on 262 sets of positioning radiographs, the average interfraction CC reproducibility (sigma) of the diaphragm and hepatic microcoils was 4.4 mm (range 3.0-6.1 mm) and 4.3 mm (range 3.1-5.7 mm), indicating a change of diaphragm and microcoil position relative to the skeleton over the course of treatment with repeat breath holds at the same phase of the respiratory cycle. The average population absolute intrafraction CC offset in diaphragm and microcoil position relative to skeleton was 2.4 mm and 2.1 mm respectively; the average absolute interfraction CC offset was 5.2 mm. Analyses of repeat CT scans demonstrated that the average intrafraction excursion of the hepatic microcoils relative to the skeleton in the CC, AP, and ML directions was 1.9 mm, 0.6 mm, and 0.6 mm respectively and the average interfraction CC, AP, and ML excursion of the hepatic microcoils was 6.6 mm, 3.2 mm, and 3.3 mm respectively. CONCLUSION: Radiotherapy using ABC for patients with intrahepatic cancer is feasible, with good intrafraction reproducibility of liver position using ABC. However, the interfraction reproducibility of organ position with ABC suggests the need for daily on-line imaging and repositioning if treatment margins smaller than those required for free breathing are a goal. PMID- 11728703 TI - Intraoperative planning and evaluation of permanent prostate brachytherapy: report of the American Brachytherapy Society. AB - PURPOSE: The preplanned technique used for permanent prostate brachytherapy has limitations that may be overcome by intraoperative planning. The goal of the American Brachytherapy Society (ABS) project was to assess the current intraoperative planning process and explore the potential for improvement in intraoperative treatment planning (ITP). METHODS AND MATERIALS: Members of the ABS with expertise in ITP performed a literature review, reviewed their clinical experience with ITP, and explored the potential for improving the technique. RESULTS: The ABS proposes the following terminology in regard to prostate planning process: *Preplanning--Creation of a plan a few days or weeks before the implant procedure. *Intraoperative planning--Treatment planning in the operating room (OR): the patient and transrectal ultrasound probe are not moved between the volume study and the seed insertion procedure. * Intraoperative preplanning- Creation of a plan in the OR just before the implant procedure, with immediate execution of the plan. *Interactive planning--Stepwise refinement of the treatment plan using computerized dose calculations derived from image-based needle position feedback. *Dynamic dose calculation--Constant updating of dose distribution calculations using continuous deposited seed position feedback. Both intraoperative preplanning and interactive planning are currently feasible and commercially available and may help to overcome many of the limitations of the preplanning technique. Dosimetric feedback based on imaged needle positions can be used to modify the ITP. However, the dynamic changes in prostate size and shape and in seed position that occur during the implant are not yet quantifiable with current technology, and ITP does not obviate the need for postimplant dosimetric analysis. The major current limitation of ITP is the inability to localize the seeds in relation to the prostate. Dynamic dose calculation can become a reality once these issues are solved. Future advances can be expected in methods of enhancing seed identification, in imaging techniques, and in the development of better source delivery systems. Additionally, ITP should be correlated with outcome studies, using dosimetric, toxicity, and efficacy endpoints. CONCLUSION: ITP addresses many of the limitations of current permanent prostate brachytherapy and has some advantages over the preplanned technique. Further technologic advancement will be needed to achieve dynamic real-time calculation of dose distribution from implanted sources, with constant updating to allow modification of subsequent seed placement and consistent, ideal dose distribution within the target volume. PMID- 11728705 TI - In regard to Sanchez-Nieto et al. IJROBP 2001;49:487-499. PMID- 11728704 TI - A practical method to achieve prostate gland immobilization and target verification for daily treatment. AB - PURPOSE: A practical method to achieve prostate immobilization and daily target localization for external beam radiation treatment is described. METHODS AND MATERIALS: Ten patients who underwent prostate brachytherapy using permanent radioactive source placement were selected for study. To quantify prostate motion both with and without the presence of a specially designed inflatable intrarectal balloon, the computerized tomography-based coordinates of all intraprostatic radioactive sources were compared over 3 consecutive measurements at 1-min intervals. RESULTS: The placement and inflation of the intrarectal balloon were well tolerated by all patients. The mean (range) displacement of the prostate gland when the intrarectal balloon was present vs. absent was 1.3 (0-2.2) mm vs. 1.8 (0-9.1) mm (p = 0.03) at 2 min respectively. The maximum displacement in any direction (anterior-posterior, superior-inferior, or right-left) when the intrarectal balloon was inflated vs. absent was reduced to < or =1 mm from 4 mm. CONCLUSIONS: Both prostate gland immobilization and target verification are possible using a specially designed inflatable intrarectal balloon. Using this device, the posterior margin necessary on the lateral fields to ensure dosimetric coverage of the entire prostate gland could be safely reduced to 5 mm and treatment could be set up and verified using a lateral portal image. PMID- 11728708 TI - Macrolide antibiotics and pulmonary inflammation. AB - Many clinically effective therapeutic agents can exhibit localized and systemic effects that are manifestly different from their intended primary pharmacological mode of action. Macrolide antibiotics such as erythromycin and its derivatives are no exception. In addition to their antibacterial action, this class of antibiotics exhibits anti-inflammatory activity in a variety of airway diseases such as asthma and diffuse panbronchiolitis that is separate and distinct from a direct antibacterial action. A variety of erythromycin derivatives have been shown to be clinically beneficial in these airway diseases. The anti-inflammatory activities of these macrolide antibiotics are becoming a research topic of intense interest. Recent work in this field has led to the understanding of the various physiological, cellular and molecular processes of the inflammatory response that are inhibited or suppressed by these compounds. This review presents a brief summary of the fascinating recent work in this active research area. PMID- 11728709 TI - The coenzyme A-dependent, non-beta-oxidation pathway and not direct deacetylation is the major route for ferulic acid degradation in Delftia acidovorans. AB - The gene loci fcs and ech, encoding feruloyl-CoA synthetase and enoyl-CoA hydratase/aldolase, respectively, are involved in the ferulic acid catabolism in Delftia acidovorans. The amino acid sequence deduced from ech exhibited 51% identity to the enoyl-CoA hydratase/aldolase from Pseudomonas sp. strain HR199, indicating that the enzyme from D. acidovorans represents a new lineage of this protein. The genes fcs and ech were expressed in Escherichia coli enabling the recombinant strain to transform ferulic acid to vanillin as revealed by photometric and HPLC analysis. An fcs deficient mutant of D. acidovorans was unable to grow on ferulic acid. The obtained data suggest that in contrast to a previous publication the biotechnologically interesting direct non-oxidative deacetylation mechanism of ferulic acid cleavage is not realized in D. acidovorans. Instead, ferulic acid degradation in D. acidovorans proceeds via a coenzyme A-dependent non-beta-oxidative pathway. PMID- 11728710 TI - MHYT, a new integral membrane sensor domain. AB - MHYT, a new conserved protein domain with a likely signaling function, is described. This domain consists of six transmembrane segments, three of which contain conserved methionine, histidine, and tyrosine residues that are projected to lie near the outer face of the cytoplasmic membrane. In Synechocystis sp. PCC6803, this domain forms the N-terminus of the sensor histidine kinase Slr2098. In Pseudomonas aeruginosa and several other organisms, the MHYT domain forms the N-terminal part of a three-domain protein together with previously described GGDEF and EAL domains, both of which have been associated with signal transduction due to their presence in likely signaling proteins. In Bacillus subtilis YkoW protein, an additional PAS domain is found between the MHYT and GGDEF domains. A ykoW null mutant of B. subtilis did not exhibit any growth alterations, consistent with a non-essential, signaling role of this protein. A model of the membrane topology of the MHYT domain indicates that its conserved residues could coordinate one or two copper ions, suggesting a role in sensing oxygen, CO, or NO. PMID- 11728711 TI - Three molecular methods to identify Salmonella enterica serotype Typhimurium DT104: PCR fingerprinting, multiplex PCR and rapid PFGE. AB - Here we report the accuracy with which three molecular techniques (PCR fingerprinting, multiplex PCR and macrorestriction profiling) distinguished Salmonella enterica Typhimurium DT104 (hereafter referred to as DT104) from other related strains of Salmonella. Each technique was tested by screening a set of 20 isolates (10 DT104, eight non-DT104 Typhimurium, one S. enterica Agona, one S. enterica Newport) and each consistently differentiated DT104 from non-DT104 isolates based on visual inspection of band patterns. The accuracy of each technique was confirmed by computer analysis. As such, these data indicate that each technique could be used to presumptively identify DT104 isolates as a precursor to phage typing. PMID- 11728712 TI - Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. AB - PCR amplification of repetitive bacterial DNA elements fingerprinting using the (GTG)(5) primer ((GTG)(5)-PCR) was proven to be useful for differentiation of a wide range of lactobacilli (i.e. 26 different (sub)species) at the species, subspecies and potentially up to the strain level. Using this rapid and reproducible genotypic technique, new Lactobacillus isolates recovered from different types of fermented dry sausage could be reliable identified at the (sub)species level. In conclusion, (GTG)(5)-PCR was found to be a promising genotypic tool for rapid and reliable speciation and typing of lactobacilli and other lactic acid bacteria important in food-fermentation industries. PMID- 11728713 TI - Potassium uptake and retention by Oceanomonas baumannii at low water activity in the presence of phenol. AB - Oceanomonas baumannii(T) (ATCC 700832) is a halotolerant bacterium capable of degrading phenol, which requires potassium in order for turgor growth to occur in minimal medium containing 5% NaCl (w/v). However, at this salinity growth can be inhibited by reduced potassium concentrations. The affinity for potassium (K(S)) was determined to be 219 microM and 408 microM for cultures utilising phenol and succinate respectively as the sole carbon source for growth. Rubidium but not caesium could substitute for potassium in alleviating growth inhibition due to potassium limitation. The effect of elevated phenol on potassium retention was studied, and it was shown that contrary to expectations, as external phenol concentration was increased the levels of intracellular potassium were significantly elevated. This observation correlated with changes in the cytoplasmic membrane, particularly the increase in the saturated:unsaturated fatty acid ratio from 0.47 to 1.44, and the decrease in the zwitterionic:anionic phospholipid ratio from 2.23 to 1.22. Both these changes promote membrane bilayer configurations and increase lipid ordering of the membrane reducing its permeability and inhibiting cation efflux. PMID- 11728714 TI - Effect of environmental and chemotactic stimuli on the activity of the Campylobacter jejuni flaA sigma(28) promoter. AB - The effect of environmental and chemotactic stimuli was assessed using a fusion between the Campylobacter jejuni flaA sigma(28) promoter and a promoterless luxCDABE cassette on pRYluxCDABE. Environmental stimuli relevant to the enteric environment were found to modulate flaA promoter activity. pH, bovine bile, deoxycholate, L-fucose and osmolarity up-regulated the flaA promoter while viscosity resulted in down-regulation. C. jejuni chemotactic effectors, including the amino acids aspartate and glutamate and the organic acids citrate, fumarate, alpha-ketoglutarate and succinate up-regulated the flaA promoter. Proline, which does not elicit a chemotactic response, up-regulated the flaA promoter. PMID- 11728715 TI - Lactococcin MMFII, a novel class IIa bacteriocin produced by Lactococcus lactis MMFII, isolated from a Tunisian dairy product. AB - A novel bacteriocin, lactococcin MMFII, produced by Lactococcus lactis MMFII isolated from a Tunisian dairy product had been identified. The bacteriocin was purified to homogeneity from fresh overnight M17 broth culture by sulfate ammonium precipitation, cation-exchange chromatography, sep-pack chromatography and two steps of reverse-phase chromatography. The purified bacteriocin was heat stable, pH resistant and protease sensitive. Its amino acid sequence, obtained by Edman degradation, revealed a 37-amino acid peptide with two cysteine residues in positions 9 and 14 and a calculated mass of 4144.6 Da. Laser desorption mass spectrometry analysis gave a molecular mass of 4142.6, suggesting the presence of a disulfide bond within the purified bacteriocin. Lactococcin MMFII contains the N-terminal YGNGV consensus motif and is active against Listeria. Thus, it belongs to the class IIa bacteriocins figuring the first example of such a bacteriocin produced by a lactococcal strain. PMID- 11728716 TI - Impact of mutations in hemA and hemH genes on pyoverdine production by Pseudomonas fluorescens ATCC17400. AB - A Pseudomonas fluorescens Tn5 mutant, with decreased production of the siderophore pyoverdine, was obtained, with the transposon inserted in the hemA gene coding for glutamyl tRNA reductase, the enzyme that catalyzes the first step of heme biosynthesis. Since this mutant was leaky, a second round of transposition was needed to obtain a second mutant completely auxotrophic for the heme precursor delta-aminolevulinate (ALA). Pyoverdine production by this mutant is ALA-dependent at concentrations above those needed to sustain growth. A transposon mutant in the hemH gene that encodes the enzyme ferrochelatase showing a characteristic red fluorescence upon UV exposure as a result of porphyrins accumulation, was obtained by selecting transconjugants on LB medium containing hemin. The DeltahemH mutant was characterized and the corresponding hemH gene sequenced. Antibodies against P. fluorescens HemH detected the protein both in soluble and membrane fractions of the wild-type and confirmed the absence of the enzyme in the mutant. The DeltahemH mutant failed to produce pyoverdine, but the production of the siderophore was restored by introduction of the Pseudomonas aeruginosa hemH gene in trans. These results indicate that de novo heme biosynthesis is needed for a normal level of siderophore pyoverdine production. PMID- 11728717 TI - Kinetoplast DNA minicircles of phloem-restricted Phytomonas associated with wilt diseases of coconut and oil palms have a two-domain structure. AB - We report the cloning and sequencing of the first minicircle from a phloem restricted, pathogenic Phytomonas sp. (Hart 1) isolated from a coconut palm with hartrot disease. The minicircle possessed a two-domain structure of two conserved regions, each containing three conserved sequence blocks (CSB). Based on the sequence around CSB 3 from Hart 1, PCR primers were designed to allow specific amplification of Phytomonas minicircles. This primer pair demonstrated specificity for at least six groups of plant trypanosomatids and did not amplify from insect trypanosomatids. The PCR results were consistent with a two-domain structure for other plant trypanosomatids. PMID- 11728718 TI - Structural and sequence diversity of the pathogenicity island of uropathogenic Escherichia coli which encodes the USP protein. AB - A total of 321 uropathogenic Escherichia coli (UPEC) strains and 12 strains of E. coli isolated from stool samples of healthy individuals, which were previously shown to be positive in colony hybridization test using the usp (encoding for the uropathogenic-specific protein) DNA probe, were examined by PCR amplification to determine the size of the usp gene and the pathogenicity island (PI). Three types of size variation were observed for the usp gene and four types for the PI. Sequencing analysis of the PIs from seven representative strains (six UPEC and one from a normal healthy individual) revealed that the usp genes can be classified into two groups, each having different sequences in the 3'-terminal region. The peptides encoded by the three open reading frames (ORFs) downstream of usp had identical 23 amino acid residues in the C-terminal region. The subregion encoding these small ORFs has a mosaic structure constituted of six segments. The positions of these segments vary from strain to strain, and in some strains, two to four segments are deleted. This indicates that rearrangements occur frequently in this region and the mosaic arrangement apparently contributes to the size variation observed in the PCR examination of the usp genes and PIs. PMID- 11728719 TI - Construction of a reporter plasmid for screening in vivo promoter activity in Francisella tularensis. AB - Francisella tularensis is a facultative intracellular bacterium that survives and multiplies inside macrophages. Here we constructed a new promoter probe plasmid denoted pKK214 by introduction of a promoter-less chloramphenicol acetyltransferase (cat) gene into the shuttle vector pKK202. A promoter library was created in F. tularensis strain LVS by cloning random chromosomal DNA fragments into pKK214. Approximately 15% of the recombinant bacteria showed chloramphenicol resistance in vitro. The promoter library was also used to infect macrophages in the presence of chloramphenicol and after two cycles of infection the library contained essentially only chloramphenicol resistance clones which shows that pKK214 can be used to monitor F. tularensis genes that are expressed during infection. PMID- 11728720 TI - Characterization of Xanthomonas oryzae pv. oryzae recX, a gene that is required for high-level expression of recA. AB - Analysis of the nucleotide sequence downstream from the Xanthomonas oryzae pv. oryzae recA gene reveals two orfs designated orfX and recX. The former has the potential to code for a 5.6 kDa protein of unknown function while the latter encodes for a putative 14.6 kDa protein with homology to RecX from various bacteria. Northern blot analysis and RT-PCR results show that recA-orfX-recX are co-regulated and arranged in an operon. A recX mutant was constructed. The mutant has no obvious growth defects or stress response defects, except that it cannot support high-level expression of recA from an expression vector. Introduction of the plasmid containing recA into the recX mutant resulted in reduced transformation efficiency and all transformants tested had mutations with reduced RecA levels. Moreover, the recX mutant has reduced basal levels of RecA. This has not been observed in other bacteria. When inactivated recX was complemented in trans, both changes were reversed. recX mutation has no effect on the regulation of the recA promoter, suggesting that its effect on the RecA level could be post transcriptional. PMID- 11728721 TI - Rapid orientated cloning in a shuttle vector allowing modulated gene expression in Bacillus subtilis. AB - An expression vector for systematic protein overproduction in Bacillus subtilis has been constructed. It derives from pDG148 and combines the main property of this vector, i.e. conditional expression of the gene in response to isopropylbeta D-thiogalactopyranoside, with (i) rapid orientated cloning by a ligation independent procedure and (ii) a ribosome binding site of high translational efficiency. When used for overproduction of several proteins in B. subtilis, this vector gave good levels of protein synthesis. PMID- 11728722 TI - The cell wall-associated serine protease PrtA: a highly conserved virulence factor of Streptococcus pneumoniae. AB - The surface-associated subtilisin-like serine protease PrtA was identified by screening a genomic expression library from Streptococcus pneumoniae using a convalescent-phase serum. In Western blot analysis two forms of PrtA were detected in whole cell lysate and a truncated form only in culture supernatant suggesting that PrtA is produced as a precursor protein, translocated to the cell surface, truncated, and released into the surroundings. A 5' fragment of the gene was found highly conserved among 78 pneumococcal isolates of clinical relevance. Immunogenicity of PrtA, limited genetic variation, and the involvement in pneumococcal virulence demonstrated in in vivo experiments might identify PrtA as a promising candidate for a protein based vaccine. PMID- 11728723 TI - Genetic analysis of a Rhodobacter capsulatus gene region involved in utilization of taurine as a sulfur source. AB - Rhodobacter capsulatus was shown to grow efficiently with taurine as sole source of sulfur. We identified a gene region exhibiting similarity to the Escherichia coli tauABC genes coding for a taurine-specific ABC transporter. The R. capsulatus tauABC genes were flanked by two putative operons (orf459-484-590 and cysE-srpI-nifS2) both reading in opposite direction relative to tauABC. Orf459 shows strong similarity to taurine:pyruvate aminotransferase (Tpa) from Bilophila wadsworthia catalyzing the initial transamination during anaerobic taurine degradation, and Orf590 exhibits clear similarity to sulfoacetaldehyde sulfo lyase from Desulfonispora thiosulfatigenes probably catalyzing the step following the taurine:pyruvate aminotransferase (Tpa) reaction, whereas nifS2 might code for a putative cysteine desulfurase. Expression of R. capsulatus tauABC and nifS2 was inhibited by sulfate, suggesting that tauABC and nifS2 might belong to the same regulon. In contrast, transcription of orf459 was not inhibited by sulfate but was induced by taurine. A tauAB deletion mutant showed significantly reduced growth compared to the wild-type with taurine as sole sulfur source in the presence of serine as a nitrogen source, whereas normal growth was observed in the presence of taurine and ammonium. Deletion of orf459-484-590 completely abolished growth with taurine/serine. Single mutations in any of the three genes resulted in the same phenotype, indicating that all three genes of this putative operon are essential for taurine sulfur utilization in the presence of serine. A model for anaerobic taurine sulfur assimilation in R. capsulatus is discussed. PMID- 11728724 TI - Novel use of guanidinium isothiocyanate in the isolation of Mycobacterium tuberculosis DNA from clinical material. AB - Nucleic acid amplification technologies offer great promise for the rapid, sensitive and specific diagnosis of tuberculosis. However, the isolation of inhibitor-free DNA from biological specimens is a bottleneck of the PCR assay. Here we describe a simple method for the isolation of PCR-amplifiable DNA of Mycobacterium tuberculosis from all types of samples of pulmonary and extrapulmonary origin tested. Briefly, it involves concentration of the bacilli by high-speed centrifugation, removal of PCR inhibitors by a wash solution containing guanidinium isothiocyanate and the release of bacterial DNA by heating in the presence of detergents and Chelex-100 resin. The entire process is accomplished within approximately 3 h. The method has been validated on 780 samples of human, bovine and guinea pig origin including sputum, cerebrospinal fluid, pulmonary fluids, pus, fine needle aspirate, tissue, blood and milk. PMID- 11728725 TI - Construction of mini-Tn5cyaA' and its utilization for the identification of genes encoding surface-exposed and secreted proteins in Bordetella bronchiseptica. AB - A mini-Tn5 transposon derivative, mini-Tn5cyaA', has been constructed. It contains a promoter-less and ribosome binding site-deficient reporter gene, encoding the catalytic domain of Bordetella pertussis adenylate cyclase toxin (CyaA'). We used this system to mutagenize B. bronchiseptica and we developed a screen for identification of mutants containing cyaA' translational fusions. This system was used to identify B. bronchiseptica genes that encode surface-exposed and secreted proteins. PMID- 11728726 TI - Involvement of the Mycobacterium tuberculosis secreted antigen SA-5K in intracellular survival of recombinant Mycobacterium smegmatis. AB - A new protein (SA-5K) secreted in culture filtrates by Mycobacterium bovis, Mycobacterium tuberculosis, and few other mycobacterial species was previously identified and purified in our laboratory. In order to evaluate the putative role of SA-5K during intracellular mycobacterial growth, in the present study the SA 5K gene was cloned and expressed in Mycobacterium smegmatis, a rapid growing non pathogenic mycobacterium which does not contain the gene for the protein. SA-5K expression in the THP-1 human macrophage cell line infected with the recombinant strain (M. smegmatis-pROL5K) was demonstrated by RT-PCR on RNA extracted from bacterial cells following 24 and 48 h of infection. Intracellular SA5K expression was associated with a higher cfu increase of M. smegmatis-pROL5K in comparison to the negative control strain (M. smegmatis recombinant for the cloning vector) (P=0.01). No significant change in SA-5K synthesis by M. smegmatis-pROL5K was observed when the recombinant strain was grown in vitro in different stress conditions such as iron deprivation, pH 4.5, presence of nitric oxide or hydrogen peroxide. The results presented in this study suggest a possible role for SA-5K in intracellular survival of recombinant M. smegmatis, though the function of the protein remains unknown. PMID- 11728727 TI - Halogenated furanones from the red alga, Delisea pulchra, inhibit carbapenem antibiotic synthesis and exoenzyme virulence factor production in the phytopathogen Erwinia carotovora. AB - The plant pathogen Erwinia carotovora regulates expression of virulence factors and antibiotic production via an N-3-oxohexanoyl-L-homoserine lactone (3-oxo-C6 HSL) dependent quorum sensing mechanism. The marine alga Delisea pulchra produces halogenated furanones known to antagonise 3-oxo-C6-HSL activity. We have tested the effects of a halogenated furanone on the production of carbapenem, cellulase and protease in E. carotovora. Despite differences in the regulatory mechanisms controlling carbapenem and exoenzyme production each was inhibited by the algal metabolite. We present evidence to suggest that the furanone dependent inhibition of carbapenem production is a result of the disruption of the 3-oxo-C6-HSL dependent expression of the carABCDEFGH operon. PMID- 11728728 TI - Protein expression by planktonic and biofilm cells of Streptococcus mutans. AB - Streptococcus mutans, a major causal agent of dental caries, functions in nature as a component of a biofilm on teeth (dental plaque) and yet very little information is available on the physiology of the organism in such surface associated communities. As a consequence, we undertook to examine the synthesis of proteins by planktonic and biofilm cells growing in a biofilm chemostat at pH 7.5 at a dilution rate of 0.1 h(-1) (mean generation time=7 h). Cells were incubated with (14)C-labelled amino acids, the proteins extracted and separated by two-dimensional electrophoresis followed by autoradiography and computer assisted image analysis. Of 694 proteins analysed, 57 proteins were enhanced 1.3 fold or greater in biofilm cells compared to planktonic cells with 13 only expressed in sessile cells. Diminished protein expression was observed with 78 proteins, nine of which were not expressed in biofilm cells. The identification of enhanced and diminished proteins by mass spectrometry and computer-assisted protein sequence analysis revealed that, in general, glycolytic enzymes involved in acid formation were repressed in biofilm cells, while biosynthetic processes were enhanced. The results show that biofilm cells possess novel proteins, of as yet unknown function, that are not present in planktonic cells. PMID- 11728729 TI - Purification of the autotransporter protein Hbp of Escherichia coli. AB - The enzyme Hbp (hemoglobin protease) of the pathogenic Escherichia coli strain EB1 has been purified to homogeneity by gel filtration chromatography. The purified protein is capable of binding heme and shows hemoglobin protease activity. Our method of purification is applicable not only to Hbp but also to other autotransporter proteins and will contribute to a better understanding of the function-structure relationship of this family of proteins. PMID- 11728730 TI - DNA from Aspergillus flavus contains 5-methylcytosine. AB - DNA from Aspergillus sp. has been reported not to contain 5-methylcytosine. However, it has been found that Aspergillus nidulans responds to 5-azacytidine, a drug that is a strong inhibitor of DNA methyltransferases. Therefore, we have re examined the occurrence of 5-methylcytosine in DNA from Aspergillus flavus by using a highly sensitive and specific method for detection of modified bases in genomic DNA comprising high-performance liquid chromatography separation of nucleosides, labeling of the nucleoside with deoxynucleoside kinase and two dimensional thin-layer chromatography. Our results show that 5-methylcytosine is present in DNA from A. flavus. We estimate the relative amounts of 5 methylcytosine to cytosine to be approximately 1/400. PMID- 11728732 TI - Heroin impurity profiling: trends throughout a decade of experimenting. AB - Heroin is still one of the most frequently abused drugs of today. All over the world, law enforcement agencies try to eradicate the illicit production and trafficking of this potent and highly addictive narcotic. To this aim, important information is provided by physical and chemical toxicological analysis of confiscated samples, with special attention for the identification and the quantification of minor components, such as the impurities related to the origin and manufacturing. By combining these data complex characterisations, i.e. impurity profiles, chemical signatures or fingerprints, can be obtained and used for comparative analysis. This review focuses on heroin impurity profiling during the 1990s, proclaimed by the United Nations as the 'Decade for Eradicating Drug Abuse'. Special attention will be given to the new trends in analytical techniques as well as in data handling strategies, so called chemometrics, to produce these profiles. The latter can be used in comparative analysis of seized heroin samples for tactical (batch-to-batch comparison) and strategic (origin determination) intelligence purposes. PMID- 11728733 TI - The potential and limitations of utilising head impact injury models to assess the likelihood of significant head injury in infants after a fall. AB - The use of engineering principles in assessing head injury scenarios is of increasing significance in investigations into suspected child abuse. A fall scenario is often given as the history for a head injury to an infant. This paper addresses the basic engineering principles and factors to be considered when calculating the severity of a head impact after free-fall. The application of head injury models (HIMs) to ascertain the forces involved in childhood head injuries from impact is also discussed. Previous studies including Duhaime et al. [J. Neurosurg. 66 (1987) 409] and Nokes et al. [Forensic Sci. Int. 79 (1995) 85] have utilised HIMs for this purpose: this paper reviews those models most widely documented.The HIM currently considered the 'state-of-the-art' is the head injury criterion (HIC) and it is suggested that this model should be utilised for assessing head impact injury in child abuse cases where appropriate. PMID- 11728735 TI - Evaluation of methyl tert-butyl ether (MTBE) as an interference on commercial breath-alcohol analyzers. AB - Anecdotal reports suggest that high environmental or occupational exposures to the fuel oxygenate methyl tert-butyl ether (MTBE) may result in breath concentrations that are sufficiently elevated to cause a false positive on commercial breath-alcohol analyzers. We evaluated this possibility in vitro by establishing a response curve for simulated breath containing MTBE in ethanol. Two types of breath-alcohol analyzers were evaluated. One analyzer's principle of operation involves in situ wet chemistry (oxidation of ethanol in a potassium dichromate solution) and absorption of visible light. The second instrument uses a combination of infrared absorption and an electrochemical sensor. Both types of instruments are currently used, although the former method represents older technology while the latter method represents newer technology.The percent blood alcohol response curve was evaluated over a breath concentration range thought to be relevant to high-level environmental or occupational exposure (0-361 microg/l). Results indicate that MTBE positively biases the response of the older technology Breathalyzer when evaluated as a single constituent or in combination with ethanol. We conclude that a false positive is possible on this instrument if the MTBE exposure is very high, recent with respect to testing, and occurs in combination with ethanol consumption. The interference can be identified on the older technology instrument by a time dependent post-reading increase in the instrument response that does not occur for ethanol alone. In contrast, the newer technology instrument using infrared and electrochemical detectors did not respond to MTBE at lower levels (0-36 microg/l), and at higher levels (>72 microg/l) the instrument indicated an "interference" or "error". For this instrument, a false positive does not occur even at high MTBE levels in the presence of ethanol. PMID- 11728734 TI - Further characterization and population data for the pentanucleotide STR polymorphism D10S2325. AB - Pentanucleotide tandem repeat markers are interesting for forensic sciences, because they may present less stutter on the electrophoretic pattern. We focused on the analysis of the DNA sequence for each allele at the pentanucleotide STR locus D10S2325 in order to understand their structures in the human genome and to construct human allelic ladder, which is necessary for forensic DNA typing. In order to evaluate the forensic applicability of D10S2325 and to construct a preliminary database, the genotype distributions and allele frequencies in three major ethnic groups were investigated. The population samples included Caucasians (Germans), Africans (African Americans), and Asians (Chinese). A total of 520 samples from unrelated individuals was analyzed by Amp-FLP. An example of each allele and new alleles were sequenced. Allele determination was carried out by comparison with a sequenced human allelic ladder made in-house. This pentanucleotide STR provided easily interpretable results. A total of 15 alleles was found in our population samples. Three new alleles were observed and named as alleles 19 and 21 based on the number of repeat motifs, while allele 19 can be divided further into two alleles, 19a and 19 according to analysis of the sequence. No evidence of deviation from Hardy-Weinberg equilibrium was observed. In 64 confirmed father/mother/child triplets no mutation event was observed. Using a maximum likelihood method, the mutation rate was indirectly estimated as 2.5 x 10(-5). These results suggest that D10S2325 is a useful marker for forensic casework and paternity analysis. PMID- 11728736 TI - Hair testing for drugs of abuse: evaluation of external cocaine contamination and risk of false positives. AB - In some laboratories hair testing may be the main method for the evaluation of individual's drug history, however, compelling evidence supports the possibility that the presence of a small amount of drug in hair can derive from external contamination. The aim of the present study is to verify if a single external contamination with a small amount of cocaine will last sufficiently long to make a contaminated subject indistinguishable from active users, and if normal washing practices together with the decontamination procedures are sufficient to completely remove the external contamination. The results obtained using the decontamination methods suggested in literature demonstrate that significant concentrations of cocaine (>1 ng/mg) and moderate quantities of benzoylecgonine (generally <0.5 ng/mg) are still detectable up to 10 weeks after contamination. These results question the reliability of hair testing. In fact, even using the most sophisticated decontamination procedures it is not possible to distinguish a drug-contaminated subject from an active user. Thus, while a negative result excludes both chronic use and "contact" with drugs, a positive result cannot and must not be interpreted as a sure sign of drug addiction, but should be further confirmed by urine analysis. PMID- 11728737 TI - The post-mortem concentration of glutamate in the structures of rat brain as an exponent of short aversive sensory stimulation preceding death. AB - The problem of post-mortem assessment of short central fear reaction preceding death has been obscure till now. The preliminary results obtained by authors have encouraged them to further research on this topic. Time of aversive sensory stimulation in the presented study was considerably decreased in comparison to the preliminary investigations. The concentration of glutamate was assayed in the selected structures of limbic system and in the cerebellum. The rats were subjected to different simultaneous modalities of sensory stimulation. The involvement of the hippocampi and the cerebellum was revealed as measured by glutamate concentration increase in the whole structures homogenates. The dominant role of mechanical stimulus has been suggested based on the measured increase of glutamate concentration in the whole cerebellum homogenate. By using the presented experimental paradigm a possible application of the biochemical assessment of human brain tissue might be developed in the future for implementing in the field of forensic pathology. The biochemical evaluation of "frozen frames" of neurotransmission can possibly help reconstruct the events which had happened just before sudden and violent death. PMID- 11728738 TI - Accidental sharp force fatalities--beware of architectural glass, not knives. AB - In a retrospective evaluation of 799 consecutive autopsies of victims of sharp force performed between 1967 and 1996 in Munster and Berlin, only 18 cases (2.3%) were classified as accidents. A typical pattern was present in 15 cases: inebriated adults (1.4-3.6g/l BAC) fell into an architectural glass surface in the form of a door or window (12 cases), an aquarium, a mirrored wardrobe or a telephone cell. Another man fell into a large drinking glass. Many victims in this group showed multiple scratches, abrasions and superficial incisions as well as one or more deep tear/cut/puncture injury. The wound margins can be clean-cut or irregular and abraded. Death was mostly caused by exsanguination except for one case of air embolism and one case of cerebral injury. The fatal injuries were produced by large and dagger-like slivers of glass, by sharp-edged fragments of glass remaining inside the frame or by a portion of glass which fell down and acted in a way similar to a guillotine. Ordinary types of flat glass were involved in all cases and it is not until the impact that sharp fragments or cutting edges are produced. So the motion of the person commonly provides the force necessary for a fatal injury. This was also true for the remaining two cases not involving architectural glass. A farmer suffered cerebral injury from a fall into the long prong of a pitch fork, and the wounding agent was a knife in only one case. A man who stated that he had fallen into the knife in his hand died from pneumonia after inadequate therapy following a single stab injury to the periphery of the left lung and liver. Accidents where the victim is killed by his own knife therefore appear to be extremely rare. PMID- 11728739 TI - Determination of opiates in postmortem bone and bone marrow. AB - Bone and bone marrow of a fatally poisoned heroin addict were analyzed by FPIA and GC-FID, immediately after death. A piece of the bone from the above case was buried for 1 year and analyzed by the same procedure. Morphine was detected in all specimens at concentrations of 195, 340 and 155 ng/g for bone marrow, bone and buried bone, respectively. A loss of 54.4% of morphine concentration was observed during 1-year burial. Such findings have potential forensic value in cases of skeletonized remains. PMID- 11728741 TI - Unidentified bodies/skulls found in Danish waters in the period 1992-1996. AB - In the period 1992-1996 a total of 89 unidentified bodies/skulls found in Danish waters were investigated at the three University Institutes of Forensic Medicine in Denmark. Males constituted 79% of the total material. Open water was the finding place in half the cases, about one-fourth was found on shore and another fourth in a harbor basin. Totally 73 bodies and one skull were identified. Danes constituted 57% of the material. Almost all non-Danes originated from countries with borders to the waters surrounding Denmark. The cause of death was in 71% stated as presumably/possibly drowning, in 5% as severe lesions (precipitation) and in 25% it could not be solved. The manner of death was in non-Danes most often stated as an accident (56%), and according to this the finding place in non Danes most frequently was the open water (66%). In Danes suicide predominated (71%) and the finding place in Danes was almost equally distributed among open water, shores and harbors, respectively. Among the totally 74 identified cases, 83% of the total material, an initial suggestion of the identity based on effects found on the body was possible in 43 cases (58%). The final identification was performed by means of forensic dental examination in total 58 cases (78%), in Danes in 93% and in non-Danes in 59%. In the remaining 16 cases one or more of the following contributed to the final identification: information about dental condition, clothing, personal papers, X-ray photos, jewelry, tattoos/scars and fingerprints. The importance of a close collaboration between the police, the forensic pathologist and the forensic dental expert in order to reach a fair number of positive identifications is emphasized. PMID- 11728740 TI - Induction of nitric oxide synthase by traumatic brain injury. AB - We investigated the dynamic induction/expression of inducible nitric oxide synthase (iNOS) using human brains made available through death by traumatic brain injury (TBI). Astrocytes, microglia, and neutrophils were identified in tissue using immunohistochemical staining with antibodies against glial fibrillary acidic protein (GFAP), MHC class II antigen, and neutrophil elastase, respectively. The localization of iNOS protein in each of these cell types was evaluated using immunohistochemistry. Within 2 days of injury, iNOS immunoreactivity was not detected. However, after 2 days, immunoreactivity was detected in the traumatized brain. The iNOS immunoreactivity was localized on neutrophils and microglia/macrophages in the areas around the tissue necrosis in the traumatized cortical hemisphere, in the deep part of the cortex and the dentate gyri of the hippocampi adjacent to the hemorrhage, and within the cytoplasm of vascular smooth muscle cell of a small artery or arteriole surrounding the injured region. This reactivity was absent after 8 days post injury.These observations confirmed the prolonged induction of iNOS within various cells in the injured brain. These responses suggest that iNOS plays a crucial role in cerebrovascular damage and/or secondary brain damage subsequent to traumatic brain injury. Furthermore, the dense nitric oxide (NO) generated by iNOS may play a role in neuronal cell death after injury. PMID- 11728742 TI - Delta(9)-THC, 11-OH-Delta(9)-THC and Delta(9)-THCCOOH plasma or serum to whole blood concentrations distribution ratios in blood samples taken from living and dead people. AB - The recreational use and abuse of Cannabis is continuously increasing in Switzerland. Cannabinoids are very often detected alone or in combination with other drugs in biological samples taken from drivers suspected of driving under the influence of drugs. Moreover, they are also frequently found in blood specimens from people involved in various medico-legal events, e.g. muggings, murders, rapes and working accidents as well. In order to assess the influence of Cannabis exposure on man behavior and performances, it is often needed to estimate the time of Cannabis use. For that purpose two mathematical models have been set up by Huestis and coworkers. These models are based on cannabinoids concentrations in plasma. Because plasma samples are rarely available for forensic determinations in our laboratory, it could be useful to assess the time laps since Cannabis use through these models from whole blood values. One prerequisite to the use of these models from whole blood values is the knowledge of the plasma to whole blood concentrations distribution ratios of cannabinoids. In this respect, the Delta(9)-THC, 11-OH-Delta(9)-THC and Delta(9)-THCCOOH concentrations were measured in plasma and whole blood taken from eight volunteers who smoke Cannabis on a regular basis. Cannabinoids levels were also determined in "serum" and whole blood samples taken from six corpses. The values of the plasma to whole blood distribution ratios were found to be very similar and their individual coefficient of variation relatively low suggesting that plasma levels could be calculated from whole blood concentrations taken into account a multiplying factor of 1.6. The data obtained postmortem suggest that the distribution of cannabinoids between whole blood and "serum" is scattered over a larger range of values than those determined from living people and that more cannabinoids (mean value of the serum/whole blood concentrations ratios=2.4) can be recovered from the "serum" fraction. The successful use of the mathematical models of Huestis and coworkers may, therefore, rely in part upon the selection of the appropriate blood sample, i.e. plasma. When plasma is not available, whole blood values could be considered with some caution taken into account a multiplying factor of 1.6 to calculate plasma concentrations from blood values. In the case of blood samples taken after death, the use of these models to assess the time of Cannabis use is not recommended. PMID- 11728743 TI - Kicking to death - forensic and criminological aspects. AB - A total of 36,274 forensic autopsies was performed in Berlin, between 1980 and 1987, including 152 cases (0.42%) in which death had been caused by blunt violence due to kicking. Data were collected on both victims and offenders, postmortem findings, causes of death and the way violence had been perpetrated. The greater part of victims and offenders had been males originating from lower social strata. Most of the victims and offenders had been in relationship with each other prior to the offence. Typical course of events: Victims and offenders, under influence of alcohol, became involved in a brawl, usually for trivial reasons, which soon led to physical fighting. When the victim had been knocked to the ground, the offender started forceful kicking. Bleeding to death and head injury were frequent causes of death. More than 50% of all offences were committed by single offenders. The diagnosis of kicking to death can at best be derived from presence of boot traces leaving shaped injuries. The trace generating boot can be identified as offending tool by means of comparative police investigation. In addition to evaluation of shoe sole profiles, there is other trace-relevant material that may be sampled from a suspected offender's footwear (skin cells, hair, blood, body tissue) and used to identify findings by DNA analysis. There may be injuries with visible patterns supporting suspicion of kicking and trampling, although conclusive confirmation can be obtained only by testimony by a witness or confession by the offender. PMID- 11728744 TI - Calculation of tension exerted on a ligature in incomplete hanging. AB - The study is devoted to one of the numerous factors (namely, body posture), which determine the strength of the neck compression in hanging. Mathematical modeling method and achievements of biomechanics were used. Human body was supposed to consist of 16 freely joined hard parts. The possibility of ligature tension calculation in any position in incomplete hanging is shown. Weight ranges for hanging of male persons in different body positions are presented: in standing posture the ligature is stretched by more than 65% of the body weight, in kneeling position by 64-74%, in sitting by 17-32%, in recumbent posture by less than 18% of the body weight. The importance of the accurate recording of a corpse posture at the place of accident is emphasized, as well as the necessity of the corpse mass evaluation. Significance of the work for medico-legal practice is illustrated by the study of the neck organocomplex trauma in 108 cases of suicidal hanging. Injuries were revealed stereomicroscopically after the complete preparation of the hyoid bone and laryngeal cartilages, preliminary fixed in the formalin solution. Trauma to the neck complex (fractures, infractions, fissures) was found in more than 64% of cases, differences in many of the groups compared are significant, P<0.05. Vulnerability index hyoid bone/thyroid cartilage was 1.75 in complete hanging of the body, 0.88 in standing position, 0.63 in kneeling, 0.33 in sitting, 0.25 in recumbent posture. Results of the study may be used for investigation of dying processes and morphology of hanging. PMID- 11728745 TI - Pressure on the neck calculated for any point along the ligature. AB - It had been experimentally shown that disfunction of the airways, major arteries and veins occurs in neck compression due to the violence of different strengths. These differences in the vulnerability of the neck anatomical structures determine the sequence and dynamics of the vital function disorders in strangulation, as well as its morphological features. The purpose of this study was to calculate the value of specific pressure P, which affects the neck along the ligature. Mathematical modeling method was used. Cylinder served the model of the human neck. It was found that P-value is directly proportional to the loop tension and inversely proportional to the radius of the curvature and neck ligature contact area. Thus, in horizontal neck encirclement the strangulation groove forms a circumference, and P-value is invariable along the loop, as in typical cases of homicidal strangulation. In oblique position of the loop it makes an ellipse with changeable curve curvature, as in most hangings. As the formulas and graphs presented show, the increasing deflection of the loop towards the transverse plane results in increase in P-amplitude, mostly at the expense of its minimal indices (up to hundred per cent) rather than maximal ones (within 20% range). At the same time there are narrow zones along the loop, where the P-value remains almost invariable irrespective of loop deflection degree. The results of the study may be useful for investigation of thanatogenesis and pathomorphology of strangulation, including its experimental modeling. PMID- 11728746 TI - Alpha-phenylethylamine in illegally produced amphetamine. AB - The possibility of simultaneous synthesis of alpha-phenylethylamine and amphetamine from mixture of acetophenone and benzylmethylketone was studied. The structures of specific impurities were predicted and these compounds were synthesized and finally found in reaction mixtures, as well as in the final product. The data collected by gas chromatography, proton and carbon magnetic resonance, Fourier transform infrared spectrometry and mass spectrometry are presented. PMID- 11728747 TI - The use of vapour phase ultra-violet spectroscopy for the analysis of arson accelerants in fire scene debris. AB - A method has been developed for the analysis of arson accelerants in fire scene debris by vapour phase ultra-violet (UV) spectroscopy. The method is rapid, inexpensive, simple to use and is sufficiently sensitive and discriminating to be of use for the analysis of crime scene samples. Application to casework samples is described. On occasion, the method offers additional information to that which can be obtained by gas chromatography-flame ionisation detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS) and represents a useful adjunct to these techniques. In addition, the method offers advantages where the use of GC-MS analysis of arson accelerants in fire scene debris is not a practical proposition. PMID- 11728748 TI - Paternity determination when the alleged father's genotypes are unavailable. AB - In paternity testing using the DNA evidence, analysis of the deficiency case when the DNA profiles of the alleged father are not available is different from that of the case with complete evidence. In this paper, we describe how to evaluate and determine the paternity in the deficiency case, by comparing the paternity indexes of the true father and the falsely non-excluded man. PMID- 11728749 TI - A fatality caused by accidental production of hydrogen sulfide. AB - A 55-year-old male Caucasian truck driver was dead at the scene after breathing hydrogen sulfide (H(2)S) produced by an accidental transfer of sodium hydrogen sulfide (NaHS) from a tanker truck to a tank containing 4% sulfuric acid (H(2)SO(4)) and iron(II) sulfate (FeSO(4)). Autopsy of the decedent's body revealed pulmonary edema and passive congestion in lungs, spleen, kidneys, and adrenal glands. Postmortem biological samples were analyzed for carbon monoxide, cyanide, ethanol, and drugs. Since a potential exposure to H(2)S was involved, blood was also analyzed for sulfide (S(2-)). The analysis entailed isolating S(2 ) from blood as H(2)S using 0.5M H(3)PO(4), trapping the gas in 0.1M NaOH, and determining the electromotive force using a sulfide ion specific electrode. Acetaminophen at a concentration of 14.3 microg/ml was found in blood, and metoprolol was detected in the blood, liver, and kidney samples. The blood S(2-) level was determined to be 1.68 microg/ml. It is concluded that the cause of death was H(2)S poisoning associated with a hazardous material accident in an industrial situation. PMID- 11728750 TI - A comparison of the characteristics of profiles produced with the AMPFlSTR SGM Plus multiplex system for both standard and low copy number (LCN) STR DNA analysis. AB - DNA STR profiles have been generated from 1 ng and low copy number (LCN) templates using 28 and 34 cycles of amplification, respectively. Characteristics which facilitate the interpretation of profiles, such as heterozygous balance, allelic dropout and stutter proportions have been quantified. We demonstrate that a reduction in DNA template coupled with an increase in amplification cycle number produces an increased rate of allelic dropout out which can be correlated to the peak areas of those alleles observed. In addition, the LCN conditions increase the degree of peak area asymmetry observed from heterozygotes and the size range of stutters. Analysis of the data allows us to develop sets of guidelines appropriate for interpreting both single and mixed DNA profiles. PMID- 11728751 TI - Data on nine STR loci used for forensic and paternity testing in the Greek Cypriot population of Cyprus. AB - Allele frequencies for the nine STRs included in the AMPFlSTR kit were obtained from a sample of 152 unrelated Greek Cypriot from the Mediterranean island of Cyprus PMID- 11728752 TI - Turkish population data with the CODIS multiplex short tandem repeat loci. AB - Allele frequencies for 13 tetrameric short tandem repeat (STR) loci, CSF1PO, D18S51, D3S1358, D21S11, D5S818, FGA, D7S820, HUMTH01, D8S1179, TPOX, D13S317, VWA, and D16S539 were determined on 198 Turkish blood samples. PMID- 11728753 TI - Korean population genetic data for eleven STR loci. AB - Allele frequency and forensic parameters for eleven STR loci were surveyed in a random sample from the Korean population. PMID- 11728754 TI - STR data for the SGM Plus loci from three Indonesian populations. AB - Allele frequencies for the 10 STRs included in the AmpFLSTR SGM Plus (Applied Biosytems) amplification kit were obtained from three populations in the Indonesian archipelago. Here, 173 unrelated Indonesian individuals were sampled, of which 44 were from the Island of Sulawesi, 44 from Sumatra and 85 from Java. PMID- 11728755 TI - Allele and haplotype frequencies for seven Y-specific STRs in an Australian Caucasian population. AB - Allele and haplotype frequencies for seven Y-Specific STR loci (DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS393 and DXYS156) were determined for 214 individuals from an Australian Caucasian population, for purposes of individuality and paternity analysis. PMID- 11728756 TI - A STR mutation in a heteropaternal twin case. AB - A heteropaternal male twin case with two men being alleged fathers was investigated as requested by the Court. Up to 37 PCR-based polymorphic DNA systems were studied in this case which was complicated by a paternal ACTBP2 mutation detected in one twin. This is the first report on a STR mutation in a double paternity case where both biological fathers were indisputably identified. The STR systems enable the resolution of these complex genetic relationships even in a case where a mutation in one STR locus was encountered. PMID- 11728757 TI - HPLC-ESI-MS/MS determination of zuclopenthixol in a fatal intoxication during psychiatric therapy. AB - The first non-suicidal fatality due to intramuscular administration of Cisordinol (zuclopenthixol, ZPT) is described. A new, rapid, and sensitive method for the determination of ZPT in postmortem specimens has been developed. High performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI MS/MS) was employed for drug confirmation and quantitation. Sample clean up was performed using a simple liquid-liquid extraction procedure. The postmortem concentration of ZPT in heart blood was 0.68 microg/ml. Furthermore, zotepine, carbamazepine, and chlorprotixene were detected in body fluids. The proposed method enables the unambiguous identification and quantitation of ZPT and other neuroleptic drugs in clinical and forensic specimens. PMID- 11728758 TI - Postmortem diffusion of drugs from the bladder into femoral venous blood. AB - We describe significantly elevated drug concentrations in the femoral venous blood due probably to postmortem diffusion from the bladder. A 16-year-old deceased male was found in a shallow ditch in winter. The estimated postmortem interval was 9 days and putrefaction was not advanced. The cardiac chambers contained fluid and coagulated blood and a small amount of buffy coat clots. Diffused hemorrhages were found in the gastric mucosa. The bladder contained approximately 600 ml of clear urine. Gas chromatographic-mass spectrometric analysis of the urine disclosed allylisopropylacetylurea (a fatty acid ureide sedative), diphenhydramine, chlorpheniramine and dihydrocodeine. The cause of death was considered to be drowning due to a drug overdose and cold exposure. The concentrations of diphenhydramine, free dihydrocodeine and total dihydrocodeine in the femoral venous blood (1.89, 3.27 and 3.30 microg/ml, respectively) were much higher than those in blood from the right cardiac chambers (0.294, 0.237 and 0.240 microg/ml, respectively). Urine concentrations of diphenhydramine, free dihydrocodeine and total dihydrocodeine were 22.6, 37.3 and 43.1 microg/ml, respectively. The stomach contained negligible amounts of diphenhydramine, free dihydrocodeine and total dihydrocodeine (0.029, 0.018 and 0.024 mg, respectively); concentrations of these drugs in the femoral muscle were 0.270, 0.246 and 0.314 microg/g, respectively. These results indicate that postmortem diffusion of diphenhydramine and dihydrocodeine from the bladder resulted in the elevated concentrations of these drugs in the femoral venous blood. Not only high urinary drug concentrations but also a large volume of urine in the bladder might accelerate the postmortem diffusion. PMID- 11728759 TI - Anomalous origin of the right coronary artery from the left sinus of Valsalva: report of two cases. AB - Anomalous origin of the right coronary artery (RCA) from the left sinus of Valsalva (LSV) is rare and has only recently been recognized as a cause of sudden cardiac death in otherwise healthy individuals. In this report, we describe two cases of anomalous origin of the RCA from the LSV. In the first case, a previously healthy 11-year-old girl died suddenly and unexpectedly. An autopsy revealed no abnormalities, except for anomalous origin of the RCA from the LSV. In the second case, a 21-year-old woman died by drowning in a bath, probably after an episode of syncope brought about by anomalous origin of the RCA from the LSV. PMID- 11728760 TI - Home administration of charcoal: can mothers administer a therapeutic dose? AB - This study evaluated whether children under 3 years of age would drink a therapeutic dose of activated charcoal (AC) in a simulated home environment. Children 13 to 35 months participated with their mothers. Children were randomly assigned to receive AC mixed with regular cola or with diet cola. Maximum time allowed to drink the AC was 30 min. A therapeutic dose was defined as 1 gm/kg or 15 g (the entire bottle) if the child weighed >15 kg. Fifteen children participated; eight received AC with regular cola; seven received AC with diet cola. Ages ranged from 13 to 30 months (average 19 months; SD 4.5 months). Eleven of 15 (73%) drank <1/2 (60 mL) of the AC. Nine of 15 (60%) drank <1/4 of the AC (30 mL). None of these children ingested a therapeutic dose. Three of the 15 (20%) drank > or =100 mL equaling a therapeutic dose. All three were in the group receiving regular cola; 62.5% (five of the eight) who had AC mixed with regular cola did not drink a therapeutic dose. The potential for failure of home AC administration needs to be considered when making the decision to recommend home stocking of AC. Mixing AC with cola does not ensure successful administration. Diet cola does not appear to be an alternative. PMID- 11728761 TI - A prospective, randomized trial of BiPAP in severe acute congestive heart failure. AB - Noninvasive positive pressure ventilation has been found to be efficacious in the setting of acute respiratory failure, specifically in chronic obstructive pulmonary disease exacerbations. Its use in congestive heart failure (CHF) is less well established. Additionally, it has been reported that there is an increase in acute myocardial infarction (AMI) rate with the use of bilevel positive pressure ventilation (BiPAP) in CHF patients. This study examined whether BiPAP decreases the intubation rate or improves cardiopulmonary parameters in severe CHF patients compared to high flow O(2) by mask (MASK), and whether there is an increase in AMI rate with the use of BiPAP. A prospective, randomized clinical trial at a county hospital teaching Emergency Department was conducted by enrolling 38 patients who were in severe CHF. Patients were randomized to receive either BiPAP or MASK in addition to adjunct therapy. Age and gender were not different between the groups. Heart rate, systolic blood pressure, diastolic blood pressure, respiratory rate, and pulse oximetry all showed no significant difference in change over time between groups, but there was a significant change over time within groups. Arterial pH, pCO(2), and pO(2) also showed no significant difference in change over time between groups, but there was a significant change over time within groups. The intubation rate for BiPAP was 23.8% (5) vs. MASK at 41.2% (7). The AMI rate was 19% (4) in the BiPAP group and 29.4% (5) in the MASK group. No true differences were detected between groups for increased oxygenation or a reduction in intubation rate. An increase in AMI rate with BiPAP was not found in this study as previously reported. This study provides support for a larger clinical trial assessing the safety and efficacy of BiPAP in acute CHF. PMID- 11728762 TI - Emergency department time for evaluation of patients discharged with a diagnosis of renal colic: unenhanced helical computed tomography versus intravenous urography. AB - The objective of this study was to determine whether patients discharged from the Emergency Department (ED) with a proven diagnosis of renal colic require less total evaluation and treatment time if unenhanced helical computed tomography (CT) rather than intravenous urography (IVU) was the diagnostic imaging study used. A retrospective review was undertaken of the medical records of 98 consecutive patients with a final diagnosis of urolithiasis or renal colic evaluated with an unenhanced helical CT scan or an IVU between January 1, 1999, and December 31, 1999. All patients were managed by Emergency Physicians and discharged from the ED. The time the patient was brought to the treatment area, the time the imaging study was ordered, and the time the patient was discharged were recorded. There were 75 patients evaluated with CT scan and 23 patients with an IVU. Patients who underwent unenhanced helical CT scan were in the ED for a mean time of 291 min [95% confidence interval (CI) 266-316] and those who had an IVU were in the ED for an average of 410 min (95% CI 340-481). Use of unenhanced helical CT scan was associated with less total time in the ED compared to IVU for patients with renal colic by a significant mean of 119 min. It is concluded that ED evaluation and treatment time of patients ultimately discharged with a proven diagnosis of renal colic is significantly less when evaluated with unenhanced helical CT scan compared to IVU. PMID- 11728763 TI - Benzyl alcohol with epinephrine as an alternative to lidocaine with epinephrine. AB - A randomized, prospective, double-blind study comparing benzyl alcohol with epinephrine, 1:100,000 (BA), and lidocaine with epinephrine, 1:100,000 (LID), as local anesthetics was carried out on adult patients with simple lacerations. The two study groups were compared for pain of infiltration (100 mm visual analog scale) and need for additional anesthesia. Pain scores were compared by a Mann Whitney Independent Rank Sum test and need for additional anesthesia by a Fishers Exact test. A total of 52 subjects (26 per group) were analyzed. The groups were similar in demographics and wound characteristics. The median pain score for BA, 7.5 mm, was less than for LID, 19.5 mm (p = 0.049). Although more patients receiving BA required additional anesthesia as compared to LID (8/26 versus 2/26), this difference did not reach statistical significance. BA is a reasonable alternative local anesthetic to LID for patients who are allergic to LID. PMID- 11728764 TI - Should transient loss of consciousness in blunt head trauma be a pre-hospital trauma triage criterion? AB - The objective of this study was to evaluate pre-hospital triage of patients with an isolated brief loss of consciousness (LOC) to a regional trauma center (RTC). Data from a 6-month period were retrospectively reviewed from an existing pre hospital data collection set. Patients were included if either they or a witness claimed a LOC, but they had regained consciousness to at least a Glasgow Coma Score (GCS) > 13 by the time the paramedics arrived. Endpoints for need for trauma center services included positive head computed tomography (CT) scan, the occurrence of emergency non-orthopedic surgery in < 6 h, admission to a surgical intensive care unit (ICU), or a length of stay (LOS) greater than 3 days for surgical evaluation. There were 655 complete records available for 275 cases of vehicular trauma and 380 cases of non-vehicular trauma. There were 170 (62%) patients in the vehicular group, and 287 (76%) in the non-vehicular group evaluated in the emergency department and discharged. In the vehicular group, only one (0.4%) patient required operative intervention in less than 6 h, three (1.1%) had a positive head CT scan, 10 (3.6%) were admitted to a surgical ICU, and four (1.5%) had a LOS > 3 days. In the non-vehicular trauma group, only one (0.3%) had surgery in < 6 h, eight (2.1%) had a positive CT scan, six (1.6%) were admitted to a surgical ICU or had a LOS > 3 days. Overall, 19 (2.9%, CI 0.018 0.045) patients met any one of the end-point criteria for trauma center utilization; however, only one patient (0.2% CI < 0.0001-0.008) required immediate neurosurgical intervention. Transient LOC, in the absence of any other American College of Surgeons (ACS) trauma triage criteria, triaged 97% of patients to a trauma center, who did not require trauma center services based on our criteria. PMID- 11728765 TI - Presentation, time to antibiotics, and mortality of patients with bacterial meningitis at an urban county medical center. AB - Our objective was to analyze the presentation, time to antibiotics, treatment, and mortality of patients with bacterial meningitis at a large urban county hospital over a 10-year period. A retrospective chart review of all patients with the diagnosis of bacterial meningitis was done. Information concerning presentation, etiologic organisms, treatment (including time to antibiotics), and outcomes were collected and analyzed. There were 165 charts reviewed with 171 total cases of bacterial meningitis. For adults with community-acquired meningitis, the mortality rate was 14%, for children it was 1.6%. Seventy-six percent of patients received antibiotics in the Emergency Department (ED) with a mean time to antibiotics of 1:08 h +/- 13 min. The rest received them as inpatients with a mean time to antibiotics of 6 +/- 9 h. The mortality rate for patients with community-acquired disease who received an Emergency Department antibiotic was 7.9%; for patients who received their antibiotics as inpatients the mortality rate was 29%. Our results indicate that the mortality rates from bacterial meningitis at our institution are lower than previously published results. Furthermore, our study supports the concept that the early administration of antibiotics in the ED may reduce mortality and may be an explanation of the lower mortality rates seen here. PMID- 11728766 TI - Delayed diagnosis of meningitis caused by beta-haemolytic group G. Streptococcus in an older woman. AB - A case of meningitis caused by group G beta-hemolytic Streptococcus (dysgalactiae, subspecies equisimilis) is reported in an 83-year-old woman. Streptococci species other than Streptococcus pneumoniae are seldom found in patients with acute bacterial meningitis, therefore, our discussion is focused on this rare organism. The question of the diagnosis of meningitis in the elderly is also addressed. PMID- 11728767 TI - Appendicitis masquerading as tumor: a case of two diagnoses. AB - Acute appendicitis remains one of the most difficult diagnoses to make in the Emergency Department. We present a puzzling and unusual case. A 47-year-old man had several hours of acute abdominal pain in the right upper quadrant, point tenderness in the right midquadrant on examination, and normal chemistries. Early appendicitis was suspected and a computed tomography (CT) scan of the abdomen was obtained. Appendicitis was not seen. What was evident was a retroperitoneal lipoma estimated to weigh 10 pounds. The general surgeon was consulted who believed that operation was necessary in light of the patient's continuing abdominal pain and the presence of the mass. Masses this large could cause pain from local compression of structures, or ischemia of the mass from outgrowing its blood supply. In surgery, a lipoma was observed that filled most of the retroperitoneum and displaced all the contents of the abdomen, including the cecum and its appendix. Also present was an acute appendicitis. On retrospective analysis of the CT scan, the appendicitis was evident but atypically located in the epigastrium. This case illustrates once again that the CT scan is a useful diagnostic adjunct for the diagnosis of new onset abdominal pain and specifically for appendicitis. PMID- 11728768 TI - Thrombolytics in infectious endocarditis associated myocardial infarction. AB - The use of thrombolytics in the management of acute myocardial infarction in eligible patients is the accepted standard of practice. We present the case of an embolic myocardial infarction in the setting of acute infectious endocarditis, treated with thrombolytics, resulting in a massive intracerebral hemorrhage and the patient's death. Historical and current literature has shown a consistent and significant incidence of concurrent intracerebral mycotic aneurysms in the setting of infectious endocarditis. Despite this, a literature review of contraindications to the use of thrombolytics rarely recognizes endocarditis as a contraindication. It is imperative that the etiology for myocardial infarction be identified; if contraindications to thrombolytic treatment exist, alternative therapeutic interventions must be pursued. This case highlights the importance of the correct etiologic diagnosis of myocardial ischemia, and increases the awareness of the significant risks of intracerebral hemorrhage associated with the use of thrombolytics in the setting of endocarditis. PMID- 11728769 TI - False-positive urine beta-HCG in a woman with a tubo-ovarian abscess. AB - Modern urine beta-human chorionic gonadotropin (HCG) assays that use enzyme linked immunosorbent assay (ELISA) technology are sensitive and specific for diagnosing pregnancy, both intrauterine and ectopic, and have become indispensable to the practice of Emergency Medicine. A urine HCG test is often relied on by the Emergency Physician as a critical component in the diagnostic regimen of a patient with a possible ectopic pregnancy. We report a case of a false-positive urine beta-HCG test in a patient with a ruptured tubo-ovarian abscess. Though false-positive pregnancy tests with tubo-ovarian abscesses have previously been reported with older methods of HCG detection, we believe that this is the first case where the pregnancy test was the modern ELISA type. The mechanism for the false-positive reaction in this case is unknown, but time may show that the ELISA test kit, like its predecessors, may occasionally give a false-positive reaction in this class of patients. PMID- 11728770 TI - An unusual case of clarithromycin associated ergotism. AB - A 41-year-old woman presented to the Emergency Department complaining of a 4-day history of worsening lower leg pain, pallor, and a sensation of coolness aggravated by exertion. Evaluation revealed severe lower extremity vasospasm. She recently had been prescribed clarithromycin for "flu-like" symptoms, and for many years had been taking a caffeine-ergotamine preparation for migraine headaches. Clarithromycin is known to interfere with ergotamine metabolism. This drug interaction is often not recognized. Ergot alkaloids are commonly used for migraine headaches and have vasoconstrictive properties. In a patient with ergotamine toxicity, these vasoconstrictive properties can lead to frank ischemia. We reviewed the literature for reports of ergotamine-associated ischemia and for reports of ergotamine toxicity caused by drug-drug interaction. PMID- 11728771 TI - Dystonia following administration of intravenous radiographic contrast material. AB - We describe a patient who experienced dystonia coincident with the administration of an i.v. contrast agent. Dystonic reactions are not well known outside of the fields of Emergency Medicine, Neurology, and Psychiatry. They have not been previously reported as a reaction to i.v. contrast material. Prompt consideration and treatment of this condition may prevent unnecessary patient discomfort and interventions. PMID- 11728772 TI - Scuba diving-induced pulmonary edema in a swimming pool. AB - SCUBA diving-induced pulmonary edema is a rare syndrome that has been previously reported to occur in cold water. We present a case of SCUBA diving-induced pulmonary edema in a 52-year-old man diving in a warm swimming pool. The pathophysiology of this syndrome is unclear, but it is unrelated to either barotrauma or decompression illness. This patient developed frank pulmonary edema while submerged, which resolved after surfacing. As with other patients who have had this syndrome, he did not have any cardiorespiratory disease. The presentation and pathophysiology of SCUBA diving-induced pulmonary edema are discussed. PMID- 11728773 TI - Emergency department D-dimer testing. AB - Since the early 1980s, much attention has been paid to the development of a biochemical marker for venous thromboembolism. D-dimers are derivatives of fibrinolysis and recently have been touted as a means of screening for thromboemoblism. We review the physiologic, pathologic, and chemical bases for this new test, and outline the specific D-dimer assays currently available. We conclude with a discussion of the clinical utility of D-dimer in the evaluation of patients with venous thromboembolism. PMID- 11728774 TI - Right lower quadrant pain. PMID- 11728775 TI - An unexpected posterior shoulder dislocation. PMID- 11728776 TI - Yo-yo impalement resulting in epidural hematoma. PMID- 11728777 TI - EMTALA: is the cure worse than the disease? PMID- 11728779 TI - Rapid sequence intubation in the field. PMID- 11728781 TI - Emergency department compression ultrasound to diagnose proximal deep vein thrombosis. PMID- 11728784 TI - Cocaine dependence: a disease of the brain's reward centers. AB - Cocaine addiction affects brain reward centers that have evolved to ensure survival. Cocaine euphoria is intensely pleasurable and results from mesolimbic dopamine (DA) neurotransmission. DA signal-receiving neurons in the nucleus accumbens synthesize endogenous opioids and project to numerous reward regions. Cocaine-induced neuroadaptations, including DA depletion, may underlie craving and hedonic dysregulation. Cue-induced craving is vigorously triggered by conditioned elements of the drug environment and associated with measurable limbic activation. Reduced frontal lobe metabolism in cocaine-addicted individuals could explain important clinical phenomena such as denial and the loss of control over limbic impulses. Cocaine addiction is rapidly progressive and associated with severe medical, psychiatric, and psychosocial consequences. Denial shields addicted individuals from their predicament and must be addressed in treatment. Lacking pharmacological options, clinicians must rely entirely on psychosocial approaches. Treatment principles, including engagement, motivational enhancement, abstinence strategies, and craving reduction are discussed in terms of biological rationales. PMID- 11728785 TI - A client-treatment matching protocol for therapeutic communities: first report. AB - The present study is the first report on a client-treatment matching protocol (CMP) to guide admissions to residential and outpatient substance abuse treatment settings. Two cohorts, a field test sample (n = 318) and cross-validation (n = 407) sample were drawn from consecutive admissions to nine geographically distributed multisetting therapeutic communities (TCs). A passive matching design was employed. Clients received the CMP on admission, but agencies were "blind" to the CMP treatment recommendation (i.e., match) and assigned clients to treatment by the usual intake procedures. Bivariate and logistical regression analyses show that positive treatment dispositions (treatment completion or longer retention in treatment)) were significantly higher among the CMP-matched clients. The present findings provide the empirical basis for studies assessing the validity and utility of the CMP with controlled designs. Though limited to TC-oriented agencies, the present research supports the use of objective matching criteria to improve treatment. PMID- 11728786 TI - Association of alcohol consumption with HIV sex- and drug-risk behaviors among drug users. AB - The relationship between alcohol use and HIV transmission is well recognized but not fully understood. In particular, the role of alcohol abuse as a mediator of HIV risk behavior among drug users is not well documented. We hypothesized that alcohol use in drug users will result in greater HIV risk-taking behavior. Participants were 354 drug users, of whom 105 were recent injection drug users. Multiple regression models were used to characterize whether measures of sexual and injection drug use HIV risk behavior were related to alcohol consumption, controlling for other potentially associated factors. We found that sexual HIV risk-taking behavior is associated with increased alcohol consumption among women (p = 0.02), with women having more risky sexual behavior than males. However, contrary to our hypothesis, there was no significant association of alcohol consumption with risky injection drug behavior. Addressing alcohol problems among drug users, particularly women, may be an important opportunity to reduce HIV sexual risk behavior among this high-risk population. PMID- 11728787 TI - Practice-based buprenorphine maintenance treatment (BMT): how do French healthcare providers manage the opiate-addicted patients? AB - France was the first country to promote the extensive use of buprenorphine for the treatment of drug-addicted subjects through the primary care system. To assess both professional commitment and patients' characteristics, all the physicians and pharmacists of a French area having prescribed/dispensed buprenorphine from 2/12/96 (the official release date) to 1/31/98 were identified from data files of the Health Insurance and then interviewed. During the first 61 weeks of buprenorphine maintenance treatment (BMT), 27.5% of physicians and 51.2% of pharmacists of that area were involved; 142 patient records were documented. Features of the clinical routines spontaneously implemented for practice-based BMT were: a high level of on-site supervised dispensation by the pharmacist (71% at treatment induction and 23% thereafter); the absence of objective measurement of illicit drug use; and a low buprenorphine dosage. These features are consistent with the lack of physicians' experience and training, and also the relatively good status of the population treated (no HIV-positives, heroin use duration averaging 4.2 +/- 3.1 years, and 81.7% with stable accommodations). Despite liberal regulations guiding BMT, a negligible proportion of cases had a "nomadic" attitude (multiple buprenorphine prescribers/deliverers). The treatment outcomes (no deaths, three drug overdoses, improvement in occupational status) are encouraging. CONCLUSION: Practice-based BMT appears to be a safe and acceptable response to moderate heroin addiction, but further training of the professionals involved and longitudinal investigations of individual outcomes are needed. PMID- 11728788 TI - Racial and ethnic differences in drug treatment entry of injection drug users in Massachusetts. AB - This article examines patterns of drug treatment entry among 28,000 adult injection drug users (IDUs), with special attention to racial and ethnic differences. The data analyzed originates from a statewide drug-treatment database covering four years. Through the use of logistic regression analysis, the study identified significant population differences between those who used detoxification only, those who used residential treatment and those who used methadone maintenance. Latino IDUs were approximately a third less likely than Caucasian IDUs to enter residential treatment and African American IDUs were half as likely as Caucasian IDUs to enter methadone maintenance treatment. This article discusses specific programmatic and research implications including the need to determine whether differential entry rates are due to access, client awareness, cultural, referral or other factors. PMID- 11728790 TI - Dual diagnosis and successful participation of adolescents in substance abuse treatment. AB - A retrospective record review of one year of admissions to a residential adolescent substance abuse treatment program (N = 91) examined the prevalence of comorbid psychiatric disorders and factors associated with successful treatment participation. Psychiatric and substance use disorders (SUD) were diagnosed by DSM-IV criteria. Successful participation was based on multiple factors assessed by the treatment team. Consistent with prior studies, there was considerable comorbidity (63.7%) with both disruptive (Attention Deficit Hyperactivity Disorder [ADHD], 11%; Conduct Disorder [CD], 24%) and other disorders (depression, 24%; adjustment disorder, 7.7%; bipolar disorder, 3.3%). Male gender was negatively associated (OR = 0.23, P = 0.019) with successful participation in univariate analyses, as was ADHD (OR = 0.18, P = 0.007). CD (OR = 0.37, P = 0.053) approached significance. Multivariate analysis reveals ADHD was significant while having CD and being male approached significance. Psychotropic medication use and other diagnoses were not associated with successful participation. It is concluded that further research on the relationship between ADHD, CD, and substance abuse treatment is needed. PMID- 11728789 TI - Excessive alcohol consumption and drinking expectations among clients in methadone maintenance. AB - Excessive alcohol consumption and related problems are common among clients in methadone maintenance treatment (MMT), yet relatively little is known about the psychological and social determinants of alcohol-related attitudes and behaviors during treatment. This study reports on the prevalence of alcohol dependence, patterns of alcohol consumption and preliminary findings about clients' beliefs that they will change their drinking behavior in the future. Data were gathered from personal interviews with 66 clients attending a MMT program in South London (some 80.5% of the eligible caseload). Forty-one percent of the overall sample met DSM-IV criteria for alcohol dependence in the past 12 months. Among clients who reported drinking in the past month (n = 50), 54% were classified as dependent, and these clients reported consuming an average of 23.5 UK standard units of absolute alcohol (188g/6.58 ounces) on a typical drinking day in the past month. Exploratory analyses suggested that expectations to change drinking behavior were predicted by subjective norms (social pressures), perceived functions of alcohol use, past drinking levels and current dose of methadone. Clinicians engaged in alcohol problems assessment and counseling during MMT could usefully examine these influences to strengthen treatment provision. PMID- 11728791 TI - Social services referral of adolescent trauma patients admitted following alcohol related injury. AB - Alcohol screening of severely injured patients should be universal. Hospitalization following alcohol-related injury provides an opportunity for intervention to reduce recidivism. This study examines the frequency of social services referral of an alcohol positive cohort of adolescent trauma patients. This was a retrospective analysis of data collected from 1994 through 1998 by the National Pediatric Trauma Registry. All patients between the ages of 12 and 17 who had a blood alcohol level (BAL) measured were analyzed. Patients receiving referral to the department of social services, family counseling, or addiction services, and those receiving any substance abuse education intervention were considered a positive referral. There were 6006 children age 12 to 17 included in the database during this five-year period, 751 of whom had a BAL measured. Of those screened, 15.5% were positive. Sixty-eight (59%) of the BAL positive patients were referred for intervention through social services. The only statistically significant predictor of referral was whether or not the patient was the operator of a vehicle involved in the motor vehicle collision. Nearly half of the adolescents in this study, who screened positive for alcohol, received no social services support. PMID- 11728792 TI - Independent prospective validation of the PaP score in terminally ill patients referred to a hospital-based palliative medicine consultation service. AB - The aim of this prospective study was to validate the Palliative Prognostic (PaP) Score in a population of hospitalized patients in Australia in order to determine its applicability in a different setting to that in which it was originally developed. Individual PaP scores were calculated for 100 terminally-ill patients consecutively referred to a palliative medicine consultation service based in a university teaching hospital. The PaP score was able to subdivide this heterogeneous patient population into three groups, the differences being highly statistically significant. Median survivals for the three groups were, respectively, 60 days (95% confidence interval 41-89 days), 34 days (25-40), and 8 days (2-11). The percentage survival at 30 days for the three groups was 66%, 54%, and 5% respectively. These data suggest that the PaP scoring system is a reasonably robust method for prognostication in advanced cancer that appears to be independent of the setting. The short survival of the third group in this study, which is consistent with the presence of a subset of gravely ill patients within the hospital setting who are referred to specialist palliative care services very late in the course of their illness, raises important issues for the care and treatment of these individuals. PMID- 11728793 TI - Nociceptive and neuropathic pain in patients with lung cancer: a comparison of pain quality descriptors. AB - Predictive validity of each word from the McGill Pain Questionnaire (MPQ) has not been investigated in relation to pain etiology. The purpose of this study was to explore differences in the words used to describe nociceptive and neuropathic pain. Patients with lung cancer (N = 123) selected words from the 78 MPQ pain quality descriptors and indicated the corresponding pain site for each word. Using only the MPQ pain location, and the cancer and treatment data abstracted from medical records, each pain site was classified as nociceptive or neuropathic (etiology). Pain etiology and quality descriptors were tested for proportional differences with sensitivity, specificity, and predictive value calculated for statistically significant descriptors. Of the 457 pain sites, 343 were classified as nociceptive (75%), 114 as neuropathic (25%). Lacerating, stinging, heavy, and suffocating were selected for a significantly larger proportion of nociceptive sites whereas throbbing, aching, numb, tender, punishing, pulling, tugging, pricking, penetrating, punishing, miserable, and nagging were selected for a larger proportion of neuropathic sites. Ten words correctly predicted 78% of the sites with 81% sensitivity to nociceptive pain and 59% sensitivity to neuropathic pain. Interestingly, several pain quality descriptors (burning, shooting, flashing, tingling, itching, and cold) previously associated with neuropathic pain did not distinguish between neuropathic and nociceptive pain. Infrequent selection of many MPQ words and lack of neurological exam data in the medical records are possible explanations for inconsistency with previous literature. Prospective studies are needed to validate pain quality descriptors for nociceptive and neuropathic types of lung cancer pain. PMID- 11728794 TI - Assessment of pain cognitions in cancer patients with chronic pain. AB - Although reports suggest that beliefs about pain play an important role in the adjustment and pain experience of non-cancer patients, the impact of cognitions on the pain experience is unknown in cancer patients. This study examined the factor structure, reliability, and validity of the Pain Cognition List Experimental version, a measure developed to assess patients' self-statements about pain and the extent to which patients are effective in dealing with the pain. The Pain Cognition List-Experimental version was administered to 313 cancer patients during hospitalization. Confirmatory factor analysis was conducted to evaluate the adequacy of fit for the original five factors. Because of a failure to replicate the factors, exploratory factor analysis was conducted, finding the factors Pain Impact, Social Comparison, Acquiescence, and Outcome Efficacy. The internal consistency for the factor Pain Impact was high (r = 0.89), while the other three factors showed low reliability. The factor Catastrophizing, usually an important factor in non-cancer patients, did not emerge. No differences were found across sex. The factors Pain Impact and Acquiescence provided evidence for concurrent validity. Patients with district nursing showed higher scores on the factors Pain Impact and Acquiescence, and higher scores on Pain Impact and Acquiescence were associated with higher pain intensity scores. This study was an attempt to evaluate pain cognitions and beliefs in cancer patients. It is concluded that the Pain Cognition List for non-cancer patients cannot be easily used in cancer pain patients and that evaluation of pain cognitions in cancer patients is useful. Health care providers evaluating pain in cancer patients with chronic pain need to be aware of the impact pain cognitions have on patients' pain experience. Although more research is needed, measuring pain cognitions should be considered. PMID- 11728795 TI - Knowledge and attitudes of Italian medical staff towards the approach and treatment of patients in pain. AB - This study is part of the project, "Towards a Pain-Free Hospital," which aims at improving the approach and treatment of the patient in pain. The objective of the study was to evaluate the knowledge and attitudes of doctors and nurses on the wards of a 1000-bed general hospital located in a rather densely-populated, industrialized area of Italy regarding the following topics: attention paid to pain, the use of analgesic drugs, and pain in children. In total, 669 nurses and 225 doctors completed a 16-item questionnaire evaluating knowledge and beliefs about pain. The overall percentage of correct answers was 61% (9.7/16). The score varied among the different wards and ranged from 48% (7.8/16) to 76% (12.3/16). Physicians scored 65% and nurses 59% (p < 0.001). The percentage of correct answers varied widely among the different items (from 30% to 96%). From these results, we conclude that there are still significant knowledge deficits and erroneous beliefs that may hamper treatment of the patient in pain. These results will help in conducting educational programs aimed at improving pain treatment in the different departments of the hospital. PMID- 11728796 TI - I get tired for no reason: a single item screening for cancer-related fatigue. AB - This study examined the criterion validity and sensitivity and specificity of a single item to rapidly screen patients in ambulatory oncology clinics for cancer related fatigue. In an effort to expand the utility of the Zung Self-Rating Depression Scale (ZSDS) as a screen for other symptoms, the utility of the single fatigue item was examined. The fatigue item reads "I get tired for no reason" and is rated on a four-point scale ranging from "none or a little of the time" to "most or all of the time." Fifty-two subjects were administered the Zung, the Functional Assessment of Cancer Therapy-Anemia (FACT-An) scale, and the Fatigue Symptom Inventory (FSI). The Zung item was highly correlated with the ZSDS (r= 0.63, p < 0.0001) and the FACT-An (r = -0.70, p < 0.0001), as well as to the individual items of the FSI, ranging from 0.41 (p < 0.003) to 0.71 (p < 0.0001). All 10 subjects considered to be depressed based on the ZSDS were also considered to fatigued on the FACT-An. Setting the ZSDS item cutoff point at level 3--"A good part of the time"--yielded a sensitivity of 78.95% and a specificity of 87.88%. It is concluded that a single item can be a fast and accurate way of screening cancer patients for fatigue to trigger additional follow-up, thus expanding the utility of a depression screening tool for problems other than the purely psychiatric. PMID- 11728797 TI - Psychological distress in amputees with phantom limb pain. AB - Research suggesting that psychological factors play a role in phantom limb pain abounds in the literature. Despite recent research suggesting that these factors exacerbate rather than cause phantom limb pain, clinicians still frequently use personality as a rationale to explain amputees' phantom limb pain. The present study aimed to examine psychological distress in a working-age population of amputees not specifically seeking help for their pain. The study was conducted in two phases. Phase 1 included 315 amputees who completed the General Health Questionnaire (GHQ). Phase 2 included a subset of the original sample who completed the Beck Depression Inventory (BDI). In Phase 1, although over 50% of the sample reported GHQ scores over the threshold used to detect "caseness," this was not related to phantom limb pain. In Phase 2 of the study, only 15% of the sample reported moderate to severe symptoms of depression. Only 4% of the variance in phantom limb pain was accounted for using the overall BDI score. When BDI items were examined individually within regression models, a number significantly predicted phantom limb pain. However, the items most related to phantom limb pain were those involved in "performance difficulties" rather than "negative affect." The present study suggests that negative affect in amputees may be related to disability rather than pain. PMID- 11728798 TI - Variation in phantom limb pain: results of a diary study. AB - Amputees experience multiple, complex problems in addition to phantom limb pain. Although studies have yielded useful data on the relationship between phantom limb pain and other variables, this research generally has evaluated only one aspect of phantom limb pain and measured it at only one time point. The present study examined ongoing phantom limb pain and associated factors prospectively through the use of hourly pain diaries that are completed over a 7-day period. The sample comprised a subset of 89 lower limb amputees taking part in a longitudinal research study. Subjects had a mean age of 46.1 years. Forty-seven per cent were female, 53% male. Subjects completed a Pain/Coping Diary that measured phantom limb pain intensity, activity levels, medication use, and alcohol use on an hourly basis over a 7-day period. It also asked subjects to list the coping strategies used on the same hourly basis over a 7-day period. The diaries highlighted the following: Phantom limb pain appears to be episodic in nature and there is great variation in its intensity. Amputees use a limited repertoire of coping strategies to deal with episodes of phantom limb pain, and of those strategies that are used, few reduce the level of pain. This variability in phantom limb pain has important implications for those involved in the care of amputees as a report of phantom limb pain at a given point in time may not reflect the amputee's overall pain experience. PMID- 11728799 TI - Epoetin alfa in cancer patients: evidence-based guidelines. AB - Anemia is a common cause of cancer-related fatigue. A systematic review of the literature was performed to establish guidelines on the use of epoetin alfa for the treatment of anemia. The evidence in support of these guidelines was selected, reviewed, and summarized by the members of the Canadian Cancer and Anemia Guidelines Development Group. The effects of epoetin alfa on quality of life (QOL) in patients with cancer were examined in 5 randomized, placebo controlled trials and 2 large, open-label, nonrandomized, community-based studies. The effects of epoetin alfa on red blood cell transfusion requirements were examined in 19 randomized controlled trials (RCTs) with 21 comparisons. All trials compared epoetin alfa to a suitable control group, examined specified outcome measures that could be analyzed, and studied patients with cancer who were receiving chemotherapy. Trials involving patients with hematologic malignancies originating in the bone marrow were excluded. Outcome measures included 1) quality of life (QOL) (as measured by scales including the Linear Analogue Self-Assessment [LASA] and the Functional Assessment of Cancer Therapy [FACT] subscales), and 2) transfusion requirements (as measured by the proportion of patients requiring transfusion and amount of transfusion). The analysis confirmed that epoetin alfa produced statistically significant and clinically relevant improvements in QOL in patients with cancer. The overall relative risk ratio for transfusion among patients receiving epoetin alfa was calculated to be 0.60 (95% Cl, 0.53-0.69; P < 0.00001), representing a 40% reduction in the proportion of patients requiring transfusion. These results support recommendations for the use of epoetin alfa in patients with cancer-related anemia. PMID- 11728800 TI - Increasing the efficacy of a celiac plexus block in patients with severe pancreatic cancer pain. AB - The purpose of this study was to evaluate the technical possibilities of placing a catheter near the celiac plexus for performance of a celiac plexus block, and to study the efficacy of repeated neurolytic celiac plexus blocks with alcohol in patients with advanced pancreatic cancer pain resistant to opioid treatment. In 12 patients, a neurolytic celiac plexus block with alcohol, administered via an indwelling celiac catheter, was performed. To evaluate the efficacy, visual analog scale scores were recorded every day. Quality of life scores were registered before and 4 weeks following the procedure. Alterations in opioid consumption, and the time between the diagnosis of pancreatic cancer and the performance of the block, were registered. All patients were followed until they died. Two patients remained without pain after the first neurolytic celiac plexus block. In all other patients a second block was administered which provided only temporary relief. Additional intermittent administration of bupivacaine through the catheter was necessary to provide adequate pain relief in these patients. Quality of life increased significantly during the treatment. Opioid consumption decreased significantly in all patients. Our study indicates that a neurolytic celiac plexus blockade with alcohol results in a significant but short-lasting analgesic effect. The use of a celiac catheter improves the long-term management of pancreatic cancer pain. PMID- 11728801 TI - Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. AB - Reactive oxygen species (ROS) are known mediators of intracellular signaling cascades. Excessive production of ROS may, however, lead to oxidative stress, loss of cell function, and ultimately apoptosis or necrosis. A balance between oxidant and antioxidant intracellular systems is hence vital for cell function, regulation, and adaptation to diverse growth conditions. Thioredoxin reductase (TrxR) in conjunction with thioredoxin (Trx) is a ubiquitous oxidoreductase system with antioxidant and redox regulatory roles. In mammals, extracellular forms of Trx also have cytokine-like effects. Mammalian TrxR has a highly reactive active site selenocysteine residue resulting in a profound reductive capacity, reducing several substrates in addition to Trx. Due to the reactivity of TrxR, the enzyme is inhibited by many clinically used electrophilic compounds including nitrosoureas, aurothioglucose, platinum compounds, and retinoic acid derivatives. The properties of TrxR in combination with the functions of Trx position this system at the core of cellular thiol redox control and antioxidant defense. In this review, we focus on the reactions of the Trx system with ROS molecules and different cellular antioxidant enzymes. We summarize the TrxR catalyzed regeneration of several antioxidant compounds, including ascorbic acid (vitamin C), selenium-containing substances, lipoic acid, and ubiquinone (Q10). We also discuss the general cellular effects of TrxR inhibition. Dinitrohalobenzenes constitute a unique class of immunostimulatory TrxR inhibitors and we consider the immunomodulatory effects of dinitrohalobenzene compounds in view of their reactions with the Trx system. PMID- 11728802 TI - Allantoin formation and urate and glutathione exchange in human muscle during submaximal exercise. AB - Seven males performed two exhaustive cycling bouts (EX1 and EX2) at a work-rate of 90% of maximal oxygen uptake, separated by 60 min. During EX1 there was a significant accumulation of urate (from 0.16 +/- 0.02 to 0.27 +/- 0.03 micromol/kg d.w.) and allantoin (from 0.39 +/- 0.05 to 0.69 +/- 0.14 micromol/kg d.w.) in the muscle. An uptake of urate was observed in early recovery from EX1 (0-9 min: 486 +/- 136 micromol; p <.05). There was no exchange of total glutathione or cysteine over the muscle either during or after exercise, and muscle and plasma total glutathione remained unaltered (p <.05). The glycogen levels were lowered by 40% at the onset of EX2, yet the level of oxidative stress in EX1 and EX2 was similar as evidenced by a similar increase in muscle allantoin in both exercise bouts. The data suggest that urate is utilized as antioxidant in human skeletal muscle and that reactive oxygen species are formed in muscle during intense submaximal exercise. No net exchange of glutathione appears to occur over the muscle either at rest, during exercise or in recovery. Moreover, when an exhaustive exercise bout is repeated with lowered glycogen levels, the level of oxidative stress is not different than that of the first bout. PMID- 11728803 TI - Molecular and kinetic study of catalase-1, a durable large catalase of Neurospora crassa. AB - Catalase-1 (Cat-1), one of the two monofunctional catalases of Neurospora crassa, increases during asexual spore formation to constitute 0.6% of total protein in conidia. Cat-1 was purified 170-fold with a yield of 48% from conidiating cultures. Like most monofunctional catalases, Cat-1 is a homotetramer, resistant to inactivation by solvents, fully active over a pH range of 4-12, and inactivated by 3-amino-1,2,4-triazole. Unlike most monofunctional catalases, Cat 1 consists of 88 kDa monomers that are glycosylated with alpha-glucose and/or alpha-mannose, is unusually stable, and is not inactivated or inhibited by hydrogen peroxide. Cat-1 was more resistant than other catalases to heat inactivation and to high concentrations of salt and denaturants. Cat-1 exhibited unusual kinetics: at molar concentrations of hydrogen peroxide the apparent V was 10 times higher than at millimolar concentrations. Inactivation of Cat-1 activity with azide and hydroxylamine was according to first order kinetics, while cyanide at micromolar concentrations was a reversible competitive inhibitor. PMID- 11728804 TI - Activation of JAK2 by the oxidative stress generated with oxidized low-density lipoprotein. AB - Atherosclerosis includes a series of cellular and molecular responses characteristic of an inflammatory disease. We provide evidence that cupric-ion oxidized LDL (CuLDL) or endothelial cell-oxidized LDL (ELDL) induced the activation by Tyr-phosphorylation of JAK2, one of the Janus kinase involved upstream of STATs in the JAK/STAT pathway of cytokine transduction. Oxidized LDL (OxLDL) also initiated STAT1 and STAT3 Tyr-phosphorylation and translocation to the nucleus, with a more marked effect for the extensively modified CuLDL. Genistein, a nonspecific Tyr-kinase inhibitor, and AG490, a specific inhibitor of JAKs, markedly prevented the CuLDL-induced enhancement of STAT1 and STAT3 Tyr phosphorylation and DNA-binding activity, suggesting that JAKs are the main kinases involved in STATs' activation by oxidized LDL. In addition, the lipid extract of CuLDL increased the intracellular levels of lipid peroxidation products and the Tyr-phosphorylation of JAK2, STAT1, and STAT3, whereas the antioxidant vitamin E prevented all these effects. These results demonstrate that OxLDL induces the activation by Tyr-phosphorylation of JAK2, STAT1, and STAT3 by generation of an intracellular oxidative stress by means of its lipid peroxidation products, and thus include JAK2 within the range of oxidative stress activated kinases. PMID- 11728805 TI - Importance of complete DNA digestion in minimizing variability of 8-oxo-dG analyses. AB - Estimates of 8-oxo-2'-deoxyguanosine (8-oxo-dG) in DNA vary at least one order of magnitude using different quantitative methods or even the same method. Our hypothesis is that an incomplete DNA hydrolysis to nucleosides by the conventional nuclease P1 (NP1) and alkaline phosphatase (AP) digestion system plays an important role in contributing to the variability of measurements using HPLC coupled with UV and electrochemical (EC) detection. We show here that factors, such as the amount of DNA, choice of enzymes, their activities, and incubation time, can affect DNA digestion and, thus, cause variability in 8-oxo dG levels. The addition of DNase I and phosphodiesterases I and II to the NP1 + AP system improves the DNA digestion by completely releasing normal nucleosides and 8-oxo-dG, thereby reducing the interday variations of 8-oxo-dG levels. Diethylenetriamine pentaacetic acid (DTPA), an iron chelator, prevented background increases of 8-oxo-dG during DNA digestion, as well as during the waiting period in the autosampler when a batch of DNA samples is analyzed by HPLC. After optimization of the DNA digestion conditions, the interday variability of 8-oxo-dG measurements using commercially available salmon testes DNA (ST DNA) were 26% over a period of 2 years. Under these optimal conditions, our laboratory variability may contribute as little as 13% to the overall variability as shown by assessment of oxidative DNA damage in a population of smokers. Based on our results, we believe that the modified DNA digestion conditions will provide much more accurate 8-oxo-dG determinations and, thus, more reliable estimates of cancer risk. PMID- 11728806 TI - Polyphosphate accumulation and oxidative DNA damage in superoxide dismutase deficient Escherichia coli. AB - Inorganic polyphosphate is a ubiquitous, linear polymer of phosphate residues linked by high-energy phosphoanhydride bonds. In response to starvation, polyP levels are increased up to 100-fold. It has been proposed that chelation of transition metals by polyP might reduce their toxicity, and that polyP accumulation is vital for survival in stationary phase. SOD-deficient E. coli is unable to survive in stationary phase. We found that deletion of the cytoplasmic SODs does not impair the cell's capability of synthesizing polyP. However, transient accumulation of polyphosphate correlated with increased resistance to H(2)O(2) and protection of DNA against oxidative damage. The reason for this protective effect of polyP is the induction of HPII catalase and DNA repair enzymes as members of the rpoS regulon. PolyP did not directly protect DNA against oxidative damage in vitro and acted as a pro-oxidant by stimulating the production of hydroxyl radical in the Fenton reaction. It is thus suggested that accumulation of poly P and rpoS induction cannot compensate for the lack of cytosolic SODs for survival in stationary phase. PMID- 11728807 TI - The effect of peroxynitrite on the catalytic activity of soluble guanylyl cyclase. AB - Soluble guanylyl cyclase (sGC) is a key enzyme of the *NO/cGMP pathway. Many cardiovascular disorders are associated with reduced *NO-mediated effects, while vascular superoxide (O(2)*(-)) production is increased. Both radicals rapidly react to peroxynitrite. We investigated whether peroxynitrite affects the activity and protein expression of sGC in intact vascular preparations. Catalytic sGC activity and expression of the sGC-beta(1) subunit was measured by conversion of radiolabeled GTP and western blot, respectively, using cytosolic extracts from rat aorta that had been incubated for 4 h with *NO/O(2)*(-) systems (devoid of free *NO) generating either 0.13 microM or 7.5 microM peroxynitrite/min. Incubation of rat aorta with 0.13 microM peroxynitrite/min had no effect. In striking contrast, incubation with 7.5 microM peroxynitrite/min resulted in a shift of the concentration-response curve obtained with a *NO donor (p =.0004) and a reduction of maximal specific activity from 3579 +/- 495 to 2422 +/- 265 pmol cGMP/mg/min (p =.036). The expression of the sGC-beta(1) subunit was unchanged. Exposure of aorta to the O(2)*(-) component had no effect, while exposure to the *NO-component reduced sGC expression to 58.8 +/- 7% (p <.001) and maximal sGC activity from 4041 +/- 992 to 1429 +/- 491 pmol cGMP/mg/min (p =.031). These data suggest that continuous generation of extracellular peroxynitrite might interfere with the *NO/cGMP signaling in vascular cells. PMID- 11728808 TI - Vitamin E kinetics in smokers and nonsmokers. AB - Does cigarette smoking increase vitamin E utilization in vivo? A trial was carried out in 6 smokers and 5 nonsmokers of comparable ages and serum lipids. Subjects consumed 75 mg each d(3)-RRR and d(6)-all rac-alpha-tocopheryl acetates (natural and synthetic vitamin E, respectively) daily for 7 d with a standardized breakfast. Fasting blood samples were drawn on days -7, -6, -5, -4, -3, -2, -1, 0, 1, 2, 3, 4, 5, 6, 7, 9, 14, 21 (negative days indicate supplementation). In both groups, plasma d(3)-alpha-tocopherol concentrations were approximately double of d(6)-alpha-tocopherol. At day 0, the %d(3) alpha-tocopherols (d(3) alpha-tocopherol/total-alpha-tocopherol x 100) were similar in both smokers and nonsmokers. Subsequently, there was a trend toward a faster exponential disappearance of the plasma %d(3) alpha-tocopherol in smokers compared with nonsmokers (0.30 +/- 0.04 compared with 0.24 +/- 0.05, p =.0565). The calculated %d(3) half-lives were 55.6 +/- 7.4 h in smokers and 72.1 +/- 17.3 h in nonsmokers (p =.0630). By day 21, the %d(3) in smokers had decreased to 1.4% +/- 0.3% while it was 2.2% +/- 0.7% (p =.0418) in the nonsmokers. These data suggest that smoking increases plasma vitamin E disappearance, but further studies are needed to confirm this finding and to assess its cause. PMID- 11728809 TI - Immunodetection of 3-nitrotyrosine in the liver of zymosan-treated rats with a new monoclonal antibody: comparison to analysis by HPLC. AB - Zymosan-induced peritonitis is associated with an increased production of reactive nitrogen oxides that may contribute to the often-observed failure of multiple organ systems in this model of acute inflammation. Quantitative biochemical evidence is provided for a marked 13-fold increase in protein-bound 3 nitrotyrosine (NTyr), a biomarker of reactive nitrogen oxides, in liver tissue of zymosan-treated rats. In order to investigate the localization of NTyr in this affected tissue, a monoclonal antibody, designated 39B6, was raised against 3-(4 hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin conjugate and its performance characterized. 39B6 was judged by competition ELISA to be approximately 2 orders of magnitude more sensitive than a commercial anti-NTyr monoclonal antibody. Binding characteristics of 39B6 were similar, but not identical, to that of a commercial affinity-purified polyclonal antibody in ELISA and immunohistochemical analyses. Western blot experiments revealed high specificity of 39B6 against NTyr and increased immunoreactivity of specific proteins from liver tissue homogenates of zymosan-treated rats. Immunohistochemical analysis of liver sections indicated a marked zymosan-induced increase in immunofluorescent staining, which was particularly intense in or adjacent to nonparenchymal cells, but not in the parenchymal cells of this tissue. Quantitative analysis of fractions enriched in these cell populations corroborated the immunofluorescent data, although the relative amounts detected in response to zymosan treatment was greatly reduced compared to whole liver tissue. These results demonstrate the high specificity of the newly developed antibody and its usefulness in Western blot and immunohistochemical analysis for NTyr, confirm the presence of NTyr by complementary methods, and suggest the possible involvement of reactive nitrogen oxides in hepatic vascular dysfunction. PMID- 11728810 TI - The stomach as a bioreactor: dietary lipid peroxidation in the gastric fluid and the effects of plant-derived antioxidants. AB - Atherosclerosis may result partly from processes that occur following food consumption and that involve oxidized lipids in chylomicrons. We investigated reactions that could occur in the acidic pH of the stomach and accelerate the generation of lipid hydroperoxides and co-oxidation of dietary constituents. The ability of dietary polyphenols to invert catalysis from pro-oxidation to antioxidation was examined. The acidic pH of gastric fluid amplified lipid peroxidation catalyzed by metmyoglobin or iron ions. Metmyoglobin catalyzed peroxidation of edible oil, resulting in 8-fold increase of hydroperoxide concentration. The incubation of heated muscle tissue in simulated gastric fluid for 2 h enhanced hydroperoxides accumulation by 6-fold to 1200 microM. In the presence of catechin or red wine polyphenols, metmyoglobin catalyzed the breakdown of hydroperoxides to zero, totally preventing lipid peroxidation and beta-carotene cooxidation. We suggest that human gastric fluid may be an excellent medium for enhancing the oxidation of lipids and other dietary constituents. The results indicate the potentially harmful effects of oxidized fats intake in the presence of endogenous catalysts found in foods, and the major benefit of including in the meal plant dietary antioxidants. PMID- 11728811 TI - Granulocyte-macrophage colony-stimulating factor ensures macrophage survival and generation of the superoxide anion: a study using a monocytic-differentiated HL60 subline. AB - A large number of constituents, such as growth factors, cytokines, and vasoregulatory molecules, contribute a network of cellular interactions to atherosclerotic lesions, and current evidence suggests that granulocyte macrophage colony-stimulating factor (GM-CSF) is one of these constituents. We conducted this study to determine whether GM-CSF has an effect on the fate and function of macrophages. We examined the effect of GM-CSF on macrophages in vitro with a highly inducible HL60 subclone (HL60/DU-1) that we recently established. HL60 cells have been reported to preserve functional GM-CSF receptors, but a GM CSF allele was rearranged and partially deleted. HL60/DU-1 cells were devoid of GM-CSF immunoreactivity and of autocrine stimulation of GM-CSF. HL60/DU-1 cells fated to die soon after terminal differentiation of macrophages by 1, 25 dihydroxy vitamin D(3) treatment. We found cell death to be mediated mainly by necrosis, not apoptosis, as confirmed by DNA fragmentation in agarose gel electrophoresis, morphological observation under a fluorescence microscope, and assay of lactate dehydrogenase release. Exogeneously administered GM-CSF rescued cells from necrotic death and caused them to survive and generate superoxide anions. We also conducted immunohistochemical analysis on an atherosclerotic human artery. Macrophages, endothelial cells, and smooth muscle cells were found to be GM-CSF positive in an atherosclerotic lesion. In summary, GM-CSF, which is produced by macrophages, endothelial cells, and smooth muscle cells, is thought to act in an autocrine and a paracrine fashion as a necrosis-inhibiting factor against arterial macrophages. This unique function may play an important role in ensuring survival and promoting function in atherosclerotic lesions. PMID- 11728812 TI - NF kappa B and AP-1 mediate transcriptional responses to oxidative stress in skeletal muscle cells. AB - The ability to induce cellular defense mechanisms in response to environmental challenges is a fundamental property of eukaryotic and prokaryotic cells. We have previously shown that oxidative challenges lead to an increase in antioxidant enzymes, particularly glutathione peroxidase (GPx) and catalase (CAT), in mouse skeletal muscle. The focus of the current studies is the transcriptional regulatory mechanisms responsible for these increases. Sequence analysis of the mouse GPx and CAT genes revealed putative binding motifs for NF kappa B and AP-1, transcriptional regulators that are activated in response to oxidative stress in various tissues. To test whether NF kappa B or AP-1 might be mediating the induction of GPx and CAT in muscle cells subjected to oxidative stress, we first characterized their activation by pro-oxidants. Electrophoretic mobility shift assays showed that oxidative stress led to increases in the DNA binding of NF kappa B in differentiated muscle cells. The NF kappa B complexes included a p50/p65 heterodimer, a p50 homodimer, and a p50/RelB heterodimer. AP-1 was also activated, but with slower kinetics than that of NF kappa B. The major component of the AP-1 complexes was a heterodimer composed of c-jun/fos. To test for redox regulation of NF kappa B- or AP-1-dependent transcriptional activation, muscle cells expressing either kappa B/luciferase or TRE/luciferase reporter constructs were subjected to oxidative stress. Pro-oxidant treatment resulted in increased luciferase activity in cells expressing either construct. To test whether NF kappa B mediates oxidant-induced increases of GPx and CAT expression, we transfected cells with either a transdominant inhibitor (I kappa B alpha) or a dominant-negative inhibitor (Delta SP) of NF kappa B. Both inhibitors blocked the induction of antioxidant gene expression by more than 50%. In summary, our results suggest that NF kappa B and AP-1 are important mediators of redox responsive gene expression in skeletal muscle, and that at least NF kappa B is actively involved in the upregulation of the GPx and CAT in response to oxidative stress. PMID- 11728813 TI - Lipid peroxidation in the rat brain after CO inhalation is temperature dependent. AB - We reported previously that 7-hydroperoxycholesterols, 7 alpha- and 7 beta hydroperoxycholest-5-en-3 beta-ol (7 alpha-OOH and 7 beta-OOH), indicated lipid peroxidation. In the present study, we measured not only 7 hydroperoxycholesterols but also oxysterols (7 alpha- and 7 beta hydroxycholesterol, 7 alpha-OH, and 7 beta-OH) and 3 beta-hydroxycholest-5-en-7 one (7-keto) in the brains of rats that underwent either a sham operation (control), hypoxia, or CO inhalation (1005 ppm) at 37 degrees C for 90 min followed by 48 h of recovery. The levels of 7-hydroperoxycholesterols, 7 beta-OH, and 7-keto were low in the hypoxia group, while the levels were unaltered in the CO group compared with the controls. Among the three groups of CO inhalation, these levels were high in the hyperthermia group (39 degrees C), and the 7 hydroperoxycholesterols were low in the hypothermia group (32 degrees C), compared with the control group. The blood O(2) saturation was almost normal in the hypothermia group, while it was similarly low in the hyperthermia and normothermia groups. The temperature-dependent lipid peroxidation in the brain after CO inhalation and recovery can not be explained by hypoxia due to CO hemoglobin formation, but may contribute to the delayed neuronal death following CO inhalation. Hypothermia may be applicable to treat patients after CO inhalation. PMID- 11728814 TI - Redox regulation of cell proliferation by pyrrolidine dithiocarbamate in murine thymoma cells transplanted in vivo. AB - Pyrrolidine dithiocarbamate (PDTC) is a synthetic compound largely used in cell biological studies and known to exert either antioxidant or pro-oxidant effects. Recently, its antitumoral activity has been proposed on the basis of its antioxidant and proapoptotic effects. In the present study, we evaluated the effect of increasing i.p. doses of PDTC on the growth of a strain of highly malignant thymoma cells inoculated in the peritoneum of inbred Balb/c mice. PDTC treatment increased the number of thymoma cells in a dose-dependent manner, enhancing the percentage of proliferating tumor cells. PDTC exerted regulatory effects on cell cycle distribution, decreasing the expression of cell cycle inhibitors. Alterations in the production of intracellular reactive oxygen species, levels of oxidized glutathione, and intracellular levels of the redox active metals iron and copper were also observed. The above results represent the first evidence that PDTC may induce in vivo cell proliferation in a murine thymoma cell model. In addition, we suggest that the ability of PDTC to bind and transport metals inside the cell and its pro-oxidant property may be factors underlying its effects on thymoma cell proliferation and cell cycle distribution. PMID- 11728815 TI - The metal-catalyzed oxidation of methionine in peptides by Fenton systems involves two consecutive one-electron oxidation processes. AB - The one-electron oxidation of methionine (Met) plays an important role in the redox reactions of Met in peptides and proteins under conditions of oxidative stress, e.g., during the metal-catalyzed oxidation of beta-amyloid peptide (beta A). However, little information is available with regard to mechanisms and product formation during the metal-catalyzed oxidation of Met. Here, we demonstrate that two-electron oxidation of Met in Fenton reactions, carried out aerobically by [Fe(II)(EDTA)](2-) and H(2)O(2) (EDTA = ethylenediaminetetra acetate) is the consequence of two consecutive one-electron transfer reactions carried out by either free or complexed hydroxyl radicals, followed by the reaction of an intermediary sulfur-nitrogen bonded radical cation (sulfuranyl radical) with O(2). The model peptide Met-Met represents an ideal substrate for these investigations as its one-electron oxidation, followed by reaction with molecular oxygen, produces unique intermediates, azasulfonium diastereomers, which can be chemically isolated before hydrolysis to sulfoxide occurs. PMID- 11728816 TI - Effect of apocynin on ozone-induced airway hyperresponsiveness to methacholine in asthmatics. AB - Apocynin is an inhibitor of NADPH oxidase present in inflammatory cells such as eosinophils and neutrophils. We investigated the effect of inhaled apocynin on ozone-induced bronchial hyperresponsiveness in vivo. Seven mild atopic asthmatics participated in a placebo-controlled, cross-over study with two exposures to O(3) at 2-week intervals. Apocynin (3 ml of 0.5 mg/ml) was inhaled 2 times before and 6 times after O(3) exposure at hourly intervals. At 36 h before and 16 h after O(3) exposure, methacholine inhalation challenge tests (Mch) were performed, and PC(20) and maximal % fall from baseline (MFEV(1)) were calculated from dose response curves. O(3)-induced change in PC(20) (Delta PC(20)) after placebo treatment was -1.94 +/- 0.39 DD (mean +/- SEM doubling dose Mch) (p =.001) and apocynin was -0.6 +/- 0.33 DD (p =.17). The difference between apocynin and placebo treatment was 1.3 DD +/- 0.42 (p =.02). O(3)-induced Delta MFEV(1) was 11.9 +/- 1.5% (p =.008) during placebo inhalation and 3.85 +/- 1.8% during apocynin (p =.47). Apocynin reduced the Delta MFEV(1) by 8.05% compared to placebo (p =.025). We conclude that apocynin markedly reduced O(3)-induced hyperreactivity for Mch as well as maximal airway narrowing. The results suggest that apocynin may have a role in preventing ozone-induced exacerbations of asthma. PMID- 11728817 TI - Evidence of oxidative stress following the viral infection of two lepidopteran insect cell lines. AB - The infection of Spodoptera frugiperda Sf-9 (Sf-9) and Trichoplusia ni BTI-Tn-5B1 4 (Tn-5B1-4) insect cell lines with Autographa californica multiple nucleopolyhedrovirus (AcMNPV) resulted in increased levels of lipid hydroperoxides and protein carbonyls. In addition, the viral infection resulted in a significant decrease in the reduced glutathione to oxidized glutathione (2GSH/GSSG) ratio. These results are all consistent with an increased level of oxidative stress as a result of the viral infection. It was also observed that the oxidative damage corresponded to reduced cell viability, i.e., the results are consistent with the premise that oxidative damage contributes to cell death. Finally, the measured intracellular activities of most of the antioxidant enzymes, specifically manganese superoxide dismutase (MnSOD), ascorbate peroxidase (APOX), and catalase (CAT, not present in Sf-9 cells), did not significantly decrease following viral infection. In contrast, the measured activity of copper-zinc superoxide dismutase (CuZnSOD) decreased in the Sf-9 and Tn-5B1-4 cells following AcMNPV infection. PMID- 11728818 TI - Upregulation of the vascular NAD(P)H-oxidase isoforms Nox1 and Nox4 by the renin angiotensin system in vitro and in vivo. AB - In different cardiovascular disease states, oxidative stress decreases the bioavailability of endothelial NO, resulting in endothelial dysfunction. An important molecular source of reactive oxygen species is the enzyme family of NAD(P)H oxidases (Nox). Here we provide evidence that the vascular Nox isoforms Nox1 and Nox4 appear to be involved in vascular oxidative stress in response to risk factors like angiotensin II (Ang II) in vitro as well as in vivo. Nox mRNA and protein levels were quantified by real-time RT-PCR and Western blotting, respectively. Nox1 and Nox4 were expressed in the vascular smooth muscle cell (VSMC) line A7r5 and aortas and kidneys of rats. Upon exposure of A7r5 cells to Ang II (1 microM, 4 h), Nox1 and Nox4 mRNA levels were increased 6-fold and 4 fold, respectively. Neither the vasoconstrictor endothelin 1 (up to 500 nM, 1-24 h) nor lipopolysaccharide (up to 100 ng/ml, 1-24 h) had any effect on Nox1 and Nox4 expression in these cells. Consistent with these observations made in vitro, aortas and kidneys of transgenic hypertensive rats overexpressing the Ren2 gene [TGR(mRen2)27] had significantly higher amounts of Nox1 and Nox4 mRNA and of Nox4 protein compared to tissues from normotensive wild-type animals. In conclusion, Nox4 and Nox1 are upregulated by the renin-angiotensin system. Increased superoxide production by upregulated vascular Nox isoforms may diminish the effectiveness of NO and thus contribute to the development of vascular diseases. Nox1 and Nox4 could be targeted therapeutically to reduce vascular reactive oxygen species production and thereby increase the bioavailability of NO. PMID- 11728819 TI - Oxidative DNA damage in human peripheral leukocytes induced by massive aerobic exercise. AB - Reactive oxygen species produced during vigorous exercise may permeate into cell nuclei and induce oxidative DNA damage, but the supporting evidence is still lacking. By using a 42 km marathon race as a model of massive aerobic exercise, we demonstrated a significant degree of unrepaired DNA base oxidation in peripheral immunocompetent cells, despite a concurrent increase in the urinary excretion of 8-hydroxy-2'-deoxyguanosine. Single cell gel electrophoresis with the incorporation of lesion-specific endonucleases further revealed that oxidized pyrimidines (endonuclease III-sensitive sites) contributed to most of the postexercise nucleotide oxidation. The oxidative DNA damage correlated significantly with plasma levels of creatinine kinase and lipid peroxidation metabolites, and lasted for more than 1 week following the race. This phenomenon may be one of the mechanisms behind the immune dysfunctions after exhaustive exercise. PMID- 11728820 TI - Dieldrin-induced oxidative stress and neurochemical changes contribute to apoptopic cell death in dopaminergic cells. AB - We examined the acute toxicity of dieldrin, a possible environmental risk factor of Parkinson's disease, in a dopaminergic cell model, PC12 cells, to determine early cellular events underlying the pesticide-induced degenerative processes. EC(50) for 1 h dieldrin exposure was 143 microM for PC12 cells, whereas EC(50) for non-dopaminergic cells was 292-351 microM, indicating that dieldrin is more toxic to dopaminergic cells. Dieldrin also induced rapid, dose-dependent releases of dopamine and its metabolite, DOPAC, resulting in depletion of intracellular dopamine. Additionally, dieldrin exposure caused depolarization of mitochondrial membrane potential in a dose-dependent manner. Flow cytometric analysis showed generation of reactive oxygen species (ROS) within 5 min of dieldrin treatment, and significant increases in lipid peroxidation were also detected following 1 h exposure. ROS generation was remarkably inhibited in the presence of SOD. Dieldrin-induced apoptosis was significantly attenuated by both SOD and MnTBAP (SOD mimetic), suggesting that dieldrin-induced superoxide radicals serve as important signals in initiation of apoptosis. Furthermore, pretreatment with deprenyl (MAO-inhibitor) or alpha-methyl-L-p-tyrosine (TH-inhibitor) also suppressed dieldrin-induced ROS generation and DNA fragmentation. Taken together, these results suggest that rapid release of dopamine and generation of ROS are early cellular events that may account for dieldrin-induced apoptotic cell death in dopaminergic cells. PMID- 11728821 TI - Kinetics of lipid peroxidation in mixtures of HDL and LDL, mutual effects. AB - In view of the proposed central role of LDL oxidation in atherogenesis and the established role of HDL in reducing the risk of atherosclerosis, several studies were undertaken to investigate the possible effect of HDL on LDL peroxidation. Since these investigations yielded contradictory results, we have conducted systematic kinetic studies on the oxidation in mixtures of HDL and LDL induced by different concentrations of copper, 2, 2'-azo bis (2-amidinopropane) hydrochloride (AAPH) and myeloperoxidase (MPO). These studies revealed that oxidation of LDL induced either by AAPH or MPO is inhibited by HDL under all the studied conditions, whereas copper-induced oxidation of LDL is inhibited by HDL at low copper/lipoprotein ratio but accelerated by HDL at high copper/lipoprotein ratio. The antioxidative effects of HDL are only partially due to HDL-associated enzymes, as indicated by the finding that reconstituted HDL, containing no such enzymes, inhibits peroxidation induced by low copper concentration. Reduction of the binding of copper to LDL by competitive binding to the HDL also contributes to the antioxidative effect of HDL. The acceleration of copper-induced oxidation of LDL by HDL may be attributed to the hydroperoxides formed in the "more oxidizable" HDL, which migrate to the "less oxidizable" LDL and enhance the oxidation of the LDL lipids induced by bound copper. This hypothesis is supported by the results of experiments in which native LDL was added to oxidizing lipoprotein at different time points. When the native LDL was added prior to decomposition of the hydroperoxides in the oxidizing lipoprotein, the lag preceding oxidation of the LDL was much shorter than the lag observed when the native LDL was added at latter stages, after the level of hydroperoxides became reduced due to their copper-catalyzed decomposition. The observed dependence of the interrelationship between the oxidation of HDL and LDL on the oxidative stress should be considered in future investigations regarding the oxidation of lipoprotein mixtures. PMID- 11728822 TI - Escape from tolerance of organic nitrate by induction of cytochrome P450. AB - The mechanism of organic nitrate tolerance is poorly defined. We studied the rat P450-catalyzed conversion of organic nitrate to nitric oxide (NO) by purified P450 isoforms relationship between P450 expression and nitrate tolerance following continuous infusion of organic nitrates in rats. The hypotensive effect of an nitroglycerin (NTG) bolus injection was abolished in rats that had been previously provided a continuous 48 h infusion of NTG. This effect was accompanied by a gradual but marked decrease in plasma and urinary nitrate levels following a peak at 18-24 h. Nitrate tolerance was reversible; the decline in the hypotensive effect and P450 levels observed after 2 d of continuous infusion was followed by restoration to control levels 2 d after cessation of the infusion. Similarly, the hypotensive action disappeared in P450-depleted, and -inhibited rats. At 48 h after infusion, NTG-induced NO generation of the vessels increased in acetone (a P450 inducer) -pretreated rats. The appearance and disappearance of P450 paralleled the conversion of organic nitrates to NO. Our observations indicate that nitrate tolerance is in large part the result of decreased P450 expression and activity. Interventions that maintain or increase P450 activity may be a strategy to provide relief from ischemic conditions in humans. PMID- 11728823 TI - Transduction of human catalase mediated by an HIV-1 TAT protein basic domain and arginine-rich peptides into mammalian cells. AB - Antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT) have been considered to have a beneficial effect against various diseases mediated by reactive oxygen species (ROS). Although a variety of modified recombinant antioxidant enzymes have been generated to protect against the oxidative stresses, the lack of their transduction ability into cells resulted in limited ability to detoxify intracellular ROS. To render the catalase enzyme capable of detoxifying intracellular ROS when added extracellularly, cell-permeable recombinant catalase proteins were generated. A human liver catalase gene was cloned and fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) and arginine-rich peptides (RRRRRRRRR) in a bacterial expression vector to produce genetic in-frame Tat-CAT and 9Arg-CAT fusion proteins, respectively. The expressed and purified fusion proteins can be transduced into mammalian cells (HeLa and PC12 cells) in a time- and dose dependent manner when added exogenously in culture medium, and transduced fusion proteins were enzymatically active and stable for 60 h. When exposed to H(2)O(2), the viability of HeLa cells transduced with Tat-CAT or 9Arg-CAT fusion proteins was significantly increased. In combination with transduced SOD, transduced catalase also resulted in a cooperative increase in cell viability when the cells were treated with paraquat, an intracellular antioxide anion generator. We then evaluated the ability of the catalase fusion proteins to transduce into animal skin. This analysis showed that Tat-CAT and 9Arg-CAT fusion proteins efficiently penetrated the epidermis as well as the dermis of the subcutaneous layer when sprayed on animal skin, as judged by immunohistochemistry and specific enzyme activities. These results suggest that Tat-CAT and 9Arg-CAT fusion proteins can be used in protein therapy for various disorders related to this antioxidant enzyme. PMID- 11728825 TI - Syndecan-4-mediated signalling. AB - The paradigm of cell surface proteoglycan function has been centered on the role of the ectoplasmic heparan sulfate (HS) chains as acceptors of a wide array of ligands, including extracellular matrix (ECM) proteins and soluble growth factors. Within this picture, the core proteins were assigned only a passive role of carrying the glycosaminoglycan (GAG) chains without direct participation in mediating outside-in signals generated by the binding of the above ligands. It appears now, however, that, side by side with the integrins and the tyrosine kinase receptors, the core proteins of the syndecan family of transmembrane proteoglycans are involved in signaling. The highly conserved tails of all the four members of the syndecan family contain a carboxy-terminal PDZ (Postsynaptic density 95, Disk large, Zona occludens-1)-binding motif, capable of forming multimolecular complexes through the binding of PDZ adaptor proteins. The cytoplasmic tail of the ubiquitously expressed syndecan-4 is distinct from the other syndecans in its capacity to bind phosphatidylinositol 4,5-bisphosphate (PIP2) and to activate protein kinase C (PKC) alpha. These properties may confer on syndecan-4 specific and unique signaling functions. PMID- 11728826 TI - The ups and downs of MEK kinase interactions. AB - MEK kinases (MEKKs) comprise a family of related serine-threonine protein kinases that regulate mitogen-activated protein kinase (MAPK) signalling pathways leading to c-Jun NH2-terminal kinase (JNK) and p38 activation, induced by cellular stress (e.g., UV and gamma irradiation, osmotic stress, heat shock, protein synthesis inhibitors), inflammatory cytokines (e.g., tumour necrosis factor alpha, TNFalpha, and interleukin-1, IL1) and G protein-coupled receptor agonists (e.g., thrombin). These stress-activated kinases have been implicated in apoptosis, oncogenic transformation, and inflammatory responses in various cell types. At present, the signalling events involving MEKKs are not well understood. This review summarises our current knowledge concerning the regulation and function of MEKK family members, with particular emphasis on those factors capable of directly interacting with distinct MEKK isoforms. PMID- 11728827 TI - Tissue distribution of GAP1(IP4BP) and GAP1(m): two inositol 1,3,4,5 tetrakisphosphate-binding proteins. AB - Two members of the GAP1 family, GAP1(IP4BP) and GAP1(m), have been shown to bind the putative second messenger Ins(1,3,4,5)P4 with high affinity and specificity, though other aspects of their behaviour suggest that in vivo, whereas GAP1(IP4BP) may function as an Ins(1,3,4,5)P4 receptor, GAP1(m) may be a receptor for the lipid second messenger PtdIns(3,4,5)P3. As a step towards clarifying their cellular roles, we describe here how we have raised and characterised antisera that are specific for the two proteins, and used these to undertake a comprehensive study of their tissue distribution. Both proteins are widely expressed, but there are several clear differences between them in the tissues that show the highest levels of expression. PMID- 11728828 TI - Neuroendocrine differentiation of the LNCaP prostate cancer cell line maintains the expression and function of VIP and PACAP receptors. AB - The molecular mechanisms involved in differentiation of prostate cancer cells to a neuroendocrine (NE) cell phenotype are not well understood. Here we used the androgen-dependent human prostate cancer cell line LNCaP to perform a systematic and broad analysis of the expression, pharmacology, and functionality of vasoactive intestinal peptide (VIP)/pituitary adenylate cyclase-activating peptide (PACAP) receptors. Reverse transcription polymerase chain reaction experiments, together with pharmacological approaches with a set of specific agonists and antagonists, demonstrated the presence of the three VIP/PACAP receptor subtypes (PAC1, VPAC1, and VPAC2 with a major role for VPAC1, acting through adenylate cyclase (AC) stimulation. An essentially similar pattern was observed by NE differentiated cells (4 days after serum deprivation) in spite of the important morphological changes observed. However, the expression of the prostate-specific antigen (PSA) decreased in NE cells (and increased again by dihydrotestosterone, DHT, treatment). The present demonstration of the induction of NE transdifferentiation in LNCaP cells by increasing concentrations of VIP adds value to previous observations on the role of cAMP in this process, an interesting topic in the comprehension of the molecular changes that are involved in the progression of prostate cancer to androgen independence. PMID- 11728829 TI - Mechanisms involved in alpha6beta1-integrin-mediated Ca(2+) signalling. AB - Contact of Jurkat T-lymphocytes with the extracellular matrix (ECM) protein laminin resulted in long-lasting alpha6beta1-integrin-mediated Ca(2+) signalling. Both Ca(2+) release from thapsigargin-sensitive Ca(2+) stores and capacitative Ca(2+) entry via Ca(2+) channels sensitive to SKF 96365 constitute important parts of this process. Inhibition of alpha6beta1-integrin-mediated Ca(2+) signalling by (1) the src kinase inhibitor PP2, (2) the PLC inhibitor U73122, and (3) the cyclic adenosine diphosphoribose (cADPR) antagonist 7-deaza-8-Br-cADPR indicate the involvement of src tyrosine kinases and the Ca(2+)-releasing second messengers D-myo-inositol 1,4,5-trisphosphate (InsP3) and cADPR. PMID- 11728830 TI - The focal adhesion targeting sequence is the major inhibitory moiety of Fak related non-kinase. AB - Focal adhesion kinase (FAK) plays an important role in integrin-mediated signal transduction pathways and its C-terminal noncatalytic domain Fak-related non kinase (FRNK), which is autonomously expressed, acts as an inhibitor of FAK. A model has been proposed where FAK and FRNK compete for an essential common binding protein. A FRNK variant in which the direct interaction with v-Crk associated tyrosine kinase substrate (CAS) was disturbed by point mutations still functioned as an inhibitor of FAK, suggesting that FRNK is unlikely to inhibit FAK by sequestering CAS. Deletion variants of FRNK within the region N-terminal to the focal adhesion targeting (FAT) sequence were still able to inhibit FAK function, indicating that this region is dispensable for the inhibitory effect of FRNK. Overexpression of a green fluorescent protein (GFP) fusion protein containing the FAT sequence delayed cell spreading and reduced FAK tyrosine phosphorylation. This indicates that the FAT sequence is the major inhibitory moiety within FRNK. PMID- 11728831 TI - The novel long PDE4A10 cyclic AMP phosphodiesterase shows a pattern of expression within brain that is distinct from the long PDE4A5 and short PDE4A1 isoforms. AB - In situ hybridisation methods were used to map the distribution of the novel long PDE4A10 isoform in the brain. PDE4A10 distribution was compared to that of the long PDE4A5 isoform and the short PDE4A1 isoform using probes specific for unique sequences within each of these isoforms. Coronal sections of the brain, taken at the level of the olfactory bulb, prefrontal cortex, striatum, thalamus, hippocampus and cerebellum, were analysed. Strongest expression of PDE4A isoforms was found in the olfactory bulb granular layer with high signals also in the piriform cortex, the dentate gyrus and the CA1 and CA2 pyramidal cells. For the two long forms, level general staining was noted throughout the striatum, thalamus and hippocampus but no signal was evident in the cerebellum. The long PDE4A10 and PDE4A5 isoforms localised to essentially the same regions throughout the brain, although PDE4A10 was uniquely expressed in the major island of Calleja. A signal for the short PDE4A1 isoform was found in regions in which the two long isoforms were both expressed, with the exception of the medial nucleus of the amygdala where weak signals for PDE4A5 and PDE4A10 were detected but PDE4A1 was absent. Uniquely, strong signals for PDE4A1 were detected in the glomerular layer of the olfactory bulb, the CA3 pyramidal cell region and the cerebellum; areas where signals for the two long forms were not evident. PDE4A transcripts for both PDE4A5 and PDE4A10 were not apparent in the brain stem and those for PDE4A1 were low. PDE4A isoforms are present in several key areas of the brain and therefore present valid targets for therapeutic interventions. Whilst the two long PDE4A isoforms show a remarkably similar distribution, in at least three regions there is clear segregation between their pattern of expression and that of the PDE4A1 short form. This identifies differential regulation of the expression of PDE4A long and short isoforms. We suggest that specific PDE4A isoforms may have distinct functional roles in the brain, indicating that PDE4A isoform-selective inhibitors may have specific therapeutic and pharmacologic properties. PMID- 11728832 TI - Clozapine: its impact on aggressive behavior among patients in a state psychiatric hospital. AB - Clozapine has shown consistent efficacy against positive symptoms of psychoses, and emerging reports indicate improvements in aggression and suicidality. This study evaluated the impact of clozapine aggression in a psychiatric hospital. Over a three year period, 137 subjects with schizophrenia or schizoaffective disorder received clozapine, of whom nearly 50% (n=69) experienced seclusion or restraint. Using a mirror-image study design, seclusion and restraint rates were computed per patient-month pre-clozapine and compared during clozapine treatment to a maximum of 12 months in either direction. The rest of the hospital not receiving clozapine served as a comparator group. Statistically significant reductions occurred in both seclusion (0.44+/-0.46 vs. 0.16+/-0.32, z=-3.91, p=0.0003) and restraint (0.34+/-0.47 vs. 0.08+/-0.23, z=-2.27, p=0.032) during clozapine treatment as compared with the pre-clozapine period. The comparator group experienced a low rate of seclusion and restraint throughout. While there are limitations to a mirror-image design, this study supports the emerging data on the benefits of clozapine for aggressive and violent patients with psychoses. Preliminary data suggests other second generation antipsychotic agents may have similar effects. PMID- 11728833 TI - Effects of risperidone on information processing and attention in first-episode schizophrenia. AB - Risperidone appears to have a beneficial effect in several areas of cognitive function in schizophrenic patients. In previous studies, however, the clinical characteristics of the subjects differed between studies, and were heterogeneous even in single study. Most of the subjects were previously exposed to different kinds of neuroleptics and some of them were treatment-resistant. The aim of this study is to evaluate the effect of risperidone on attention and information processing in a homogeneous subgroup of schizophrenic patients, i.e. drug-naive first-episode inpatients. In the patient group (n=17), cognitive tests and clinical assessments were performed before and after 8 weeks of risperidone treatment. The same cognitive tests were administered to the control group (n=24). The delay between test and retest was 8 weeks. Before treatment, the patient group performed significantly worse than control group on the tests measuring continuous attention, vigilance, and the speed of information processing. After treatment (the average dose of risperidone was 7.3mg/day), in spite of significant improvement of the clinical symptoms, the patients did not show any significant improvement or worsening in most of the items of these tests. The control group did not show any practice effect. These results suggest that first-episode schizophrenic patients have deficits in sustained attention and vigilance to visual stimuli, as well as the speed of information processing to visual and auditory stimuli, and these deficits are unrelated to clinical symptoms and remain stable during the early phase of treatment. This study did not receive pharmaceutical company financial support. PMID- 11728834 TI - Effects of the 5HT antagonist cyproheptadine on neuropsychological function in chronic schizophrenia. AB - This study tests the hypothesis that the ability of atypical neuroleptics to improve negative symptoms is due to 5HT-receptor antagonism and enhanced frontal lobe function. We investigated the effects of cyproheptadine (a 5HT2 antagonist) on neuropsychological tests of frontal lobe functions in chronic schizophrenic patients. Eighteen stable schizophrenic patients on depot neuroleptic medication participated in a 4-week double blind crossover study. Outcome measures were clinical symptoms rating scales, neuropsychological tests (verbal fluency, Stroop colour word task, trail making) and antisaccade eye movements. During the cyproheptadine phase statistically significant improvement was seen on Stroop colour word task, verbal fluency and Trail B tests. The ability to suppress reflexive eye movement to a target light in an anti saccade task was also significantly enhanced. The patients had low clinical ratings of negative symptoms and they were unaffected by cyproheptadine. The results indicate that 5HT2C receptors selectively modulate speed and motor control mechanisms related to frontal lobe functions but this was not associated with changes in symptoms. PMID- 11728835 TI - Fluphenazine plasma level monitoring for patients receiving fluphenazine decanoate. AB - BACKGROUND: Finding a dose of an antipsychotic for maintenance therapy that is both safe and effective can be difficult because clinicians are unable to titrate dose against clinical response in patients who are already stable. Therapeutic monitoring of antipsychotic plasma levels has the potential for helping clinicians in dosage selection. With this in mind, we evaluated the usefulness of monitoring fluphenazine plasma levels for patients with schizophrenia who were receiving maintenance treatment with fluphenazine decanoate. METHOD: Thirty-one patients with schizophrenia were randomly assigned to low, medium, or high (0.1 0.3, 0.3-0.6, 0.6-1.0 ng/ml) plasma levels of fluphenazine. The dose of fluphenazine decanoate was adjusted in order to maintain patients in their assigned range. Side effects, psychopathology, and psychotic exacerbations were measured during the year following randomization. RESULTS: All of the psychotic exacerbations occurred during the first eight weeks following randomization, before patients had adequate time to reach their plasma level assignments. We did not find a relationship between plasma levels of fluphenazine and clinical outcomes or side effects. CONCLUSION: Our results do not provide support for the usefulness of monitoring fluphenazine plasma levels for patients receiving fluphenazine decanoate. PMID- 11728836 TI - A comparative profile analysis of neuropsychological functioning in patients with schizophrenia and bipolar psychoses. AB - Evidence for neuropsychological deficits in schizophrenia is substantial whereas evidence for the specificity of dysfunction is relatively sparse. To assess specificity, we compared neuropsychological function in patients with chronic schizophrenia, patients with chronic psychotic bipolar disorder and normal controls. Groups were comparable on age, ethnicity and expected intellectual ability (based on single word reading). Patients with schizophrenia and bipolar psychoses were also relatively similar on age at onset and number of hospitalizations. Using multivariate analyses of variance with sex and parental SES as covariates (our primary analyses), patients with schizophrenia were significantly more impaired than controls on seven of eight neuropsychological functions (all but verbal ability), and were significantly more impaired than bipolar patients on abstraction, perceptual-motor speed and vigilance. Bipolar patients were significantly impaired compared to controls on declarative verbal memory, and showed moderate-to-large effect size decrements on abstraction, perceptual-motor speed and vigilance. Results were not attenuated when IQ was controlled, which was significantly lower in patients with schizophrenia. Analyses indicated that the two psychiatric groups had similar profile patterns, but that patients with schizophrenia had a more severe impairment than patients with bipolar psychoses. Further research is required to determine whether similar mechanisms underly the neurocognitive deficits in these disorders. PMID- 11728837 TI - De-coupling of cognitive performance and cerebral functional response during working memory in schizophrenia. AB - Working memory dysfunction is considered to be fundamental to the cognitive and clinical features evident in schizophrenia. Functional neuroimaging studies have begun to elucidate the neurobiological basis of such deficits, however, interpretation of these studies may be confounded by performance impairment, when the cognitive load exceeds the limited response capacity of patients with schizophrenia. In this study, patients were pre-selected on the basis of intact performance on a relatively low-load verbal working memory task, in order to mitigate against performance confounds. Subjects included 20 right-handed male subjects with chronic schizophrenia, and 20 right-handed, age-matched, male healthy controls, without personal or familial psychiatric history. All subjects underwent fMRI scanning whilst performing a verbal n-back task. There were no significant between-group differences in target identification; the patient group showed a significantly increased mean response latency. Both groups demonstrated robust fronto-parietal activation. In the control subjects, the power of functional response was positively correlated with reaction time in bilateral posterior parietal cortex, however, this coupling of behavioural performance and cerebral response was not evident in the patients. This deficit, apparent within the performance capacity of the patients, may represent a fundamental abnormality in schizophrenia, and may compromise performance at higher cognitive loads. PMID- 11728838 TI - A new statistical method for testing hypotheses of neuropsychological/MRI relationships in schizophrenia: partial least squares analysis. AB - We applied partial least squares (PLS) as a novel multivariate statistical technique to examine neuropsychological correlates of magnetic resonance imaging (MRI) measures of brain volumes in a well studied sample of 15 male patients with chronic schizophrenia. In the current study, because the total number of measures far surpassed the total number of subjects, extant multivariate techniques such as canonical correlation could not be used to examine relationships among simultaneous measures of MRI and neuropsychology. Moreover, because MRI measures were expected to be highly inter-correlated, as would be neuropsychological test scores, extant multivariate statistical techniques would be substantially limited because they typically assume statistical independence among sets of measures. PLS, on the other hand, proved to be especially well suited to examining the relationships among function and anatomy measures in this sample, where statistically significant relationships were demonstrated that were entirely consistent with prior studies using univariate correlation techniques. In particular, statistically significant relationships emerged among sets of MRI temporal lobe measures and neuropsychological tests of verbal memory and categorization as well as among MRI frontal measures and neuropsychological tests of working memory. PMID- 11728839 TI - Spatio-temporal disintegration of visual perception in schizophrenia as revealed by a novel cognitive task, the Searchlight Test. AB - We developed a novel computer-assisted psychological test (the Searchlight Test) in order to explore 'cognitive fragmentation' and its relation to thought disorder in schizophrenic patients. Participants were instructed to search geometric figures through a small hole (3 cm in diameter) on a display monitor by moving a mouse device and to reconstruct an image of the whole from the temporally and spatially fragmented visual stimuli.The cognitive function measures in 24 schizophrenic patients and 14 normal controls showed that the error rates of the recall task (drawing the figure from memory) and the recognition task (selecting the correct figure from several similar figures) were significantly higher in schizophrenic patients than in normal controls. The error rates of both tasks significantly correlated with disorganization syndrome in schizophrenia, but not with reality distortion or with psychomotor poverty syndrome.The present data obtained with the Searchlight Test suggest the spatio temporal disintegration of visual perception in schizophrenia and its correlation with disorganization. This test appears to provide a useful tool for studying the pathophysiology of disorganization syndrome, thought disorder and the related cognitive dysfunction of schizophrenia. PMID- 11728840 TI - Deficits of spatial working memory in chronic schizophrenia. AB - This study tests the hypothesis that deficits of spatial working memory are an inherent trait in schizophrenic patients, regardless of their clinical status. The participants were 22 chronic schizophrenic patients, 11 schizophrenic patients in remission, and 22 normal adults. All participants were asked to track the outlines of a square presented on a computer screen in a clockwise direction while moving their eyes at their preferred rate and then to reproduce the square on a blank computer screen (Task A). Thereafter, the computer only displayed the four corners of the same square. The corners were displayed sequentially in a clockwise direction, one at a time at equal intervals. They were asked first to track the four corners of the square and then to reproduce the square on a blank computer screen (Task B). Chronic schizophrenic patients reproduced tasks A and B with significantly greater distortions than normal controls. The extent of their distortions was significantly greater during the reproduction of Task A than Task B. By contrast, schizophrenic patients in remission reproduced both Tasks A and B with the same accuracy as normal controls. On the basis of these findings it is concluded that the chronic schizophrenic patients have a significant deficit of spatial working memory, but that this deficit can be ameliorated by cognitive interventions and is therefore not an immutable trait in schizophrenia. PMID- 11728841 TI - Nonverbal social perception and symptomatology in schizophrenia. AB - This study examined the relationship between nonverbal social perception and symptomatology in schizophrenia. Schizophrenic patients (n=28) and normal controls (n=28) were given the profile of nonverbal sensitivity test (PONS). Patients' symptoms were rated with the brief psychiatric rating scale (BPRS). Schizophrenic subjects performed significantly more poorly than normal subjects in their ability to decode nonverbal cues. In addition, normal subjects improved their accuracy significantly more than patients when provided with additional cues per scene to decode. The patients' total PONS score was not significantly correlated with the BPRS summary scores for positive or negative symptoms, but was significantly correlated with the individual positive symptom of conceptual disorganization. Subjects classified as having paranoid schizophrenia (n=11) on the basis of chart diagnoses performed significantly better on the PONS than did undifferentiated schizophrenic subjects (n=13). The discussion reviews how these results contribute to our understanding of social deficits in schizophrenia. PMID- 11728842 TI - Executive/attentional cognitive functions in schizophrenic patients and their parents: a preliminary study. AB - The aim of this study was to determine whether executive/attentional cognitive performances could be considered as markers of vulnerability to schizophrenia. The Stroop Color Word and fluency tests were significantly impaired in schizophrenic patients and their parents compared to controls matched on age and sex while performances on Nelson's Modified Card Sorting Test and the Trail Making Test did not differ. The impairments on the Stroop and fluency could be considered as endophenotypic markers of schizophrenia. PMID- 11728843 TI - Reduced binocular depth inversion in schizophrenic patients. AB - Binocular depth inversion represents an illusion of visual perception, serving to invert the perception of implausible hollow objects, e.g. a hollow face into a normal face. Such inversion occurs frequently, especially when objects with a high degree of familiarity (e.g. photographs of faces) are displayed. Under normal conditions, cognitive factors apparently override the binocular disparity cues of stereopsis. This internal mechanism--a kind of "censorship" of perception balancing "top-down" and "bottom-up" processes of perception--appears to be disturbed in psychotic states. The clinical and neuropsychological performance of schizophrenic patients was assessed using the Brief Psychiatric Rating Scale (BPRS), the Positive And Negative Symptoms Scale (PANSS), the Clinical Global Impression Scale (CGI), the Mehrfach-Wahlwortschatz Intelligence Test (MWT-B) and the binocular depth inversion test (BDIT) using pictures with a high degree of familiarity. In schizophrenic patients, the performance in the BDIT differed significantly from healthy controls and from patients with major depression. The schizophrenic patients were more veridical in their judgements in the BDIT. During antipsychotic treatment, BPRS and PANSS scores improved and the inversed faces were seen as more illusionary, driven by an increase in top-down processing. At the end of treatment, there was no significant difference between the patient group and the healthy controls in the score of binocular depth inversion. These findings suggest that testing of binocular depth inversion can detect specific dysfunctions in visual perception and might be useful as a state marker for psychotic states. PMID- 11728844 TI - Latent inhibition deficits in high-schizotypal normals: symptom-specific or anxiety-related? AB - Latent inhibition (LI) is the phenomenon in which subjects who have repeatedly experienced an irrelevant stimulus perform more poorly on a new learning task with that stimulus than with a novel stimulus, presumably because of a decline in stimulus-specific attention. The present article reviews the literature on LI deficits in high-schizotypal normal subjects and schizophrenic patients. Although LI-deficits have been thought to be specific to these groups, evidence is presented that the effects may be related to the anxiety components of high schizotypality and related pathologies. PMID- 11728845 TI - Trauma, PTSD, and the course of severe mental illness: an interactive model. AB - Traumatic life events, as defined by DSM-IV, are common among persons with severe mental illnesses (SMI) such as schizophrenia. Limited evidence suggests concomitantly high rates of posttraumatic stress disorder (PTSD) in this population. However, conceptual models do not exist for understanding the interactions between trauma, PTSD, and SMI. We propose a model, which is an extension of the stress-vulnerability model, in which PTSD is hypothesized to mediate the negative effects of trauma on the course of SMI. Our model posits that PTSD influences psychiatric disorders both directly, through the effects of specific PTSD symptoms including avoidance, overarousal, and re-experiencing the trauma, and indirectly, through the effects of common correlates of PTSD such as retraumatization, substance abuse, and difficulties with interpersonal relationships. We discuss the evidence supporting this model, and consider several intervening variables that are hypothesized to moderate the proposed relationships between PTSD and SMI, including social support, coping and competence, and antisocial personality disorder. Theoretical and clinical implications of the model are considered, as well as several methodological and nosological issues. We conclude with a brief discussion of directions for future research aimed at evaluating components of the model. PMID- 11728846 TI - First vs multiple episode schizophrenia: two-year outcome of intermittent and maintenance medication strategies. AB - Results of studies on intermittent neuroleptic treatment strategies in first episode (FE) schizophrenia have not been published. Aims of the present study were to elucidate the comparative efficacy of prodrome-based neuroleptic intervention in first vs multiple episode (ME) schizophrenia. As to the methods, three randomly assigned open neuroleptic treatment strategies were compared over 2 years in 363 schizophrenic outpatients (115 FE, 248 ME; ICD-9, RDC): maintenance medication vs two intermittent medication strategies (prodrome-based intervention and crisis intervention). Concerning relapse prevention, the results demonstrate that ME patients seemed to profit most from maintenance medication compared to both intermittent treatments, whereas FE patients did equally well under maintenance medication and prodrome-based intervention treatment. Psychopathology, social adjustment, subjective well-being, and side-effects after two years did not differ significantly between the FE and ME patients irrespective of treatment strategy. Concerning treatment adherence, FE patients complied better with prodrome-based intervention than with maintenance medication. Cumulative neuroleptic dosage was lowest in FE patients under intermittent treatment. In conclusion, maintenance medication is the best strategy for relapse prevention in ME patients. In FE patients, prodrome-based intermittent intervention seems to be equivalent or even better with respect to compliance and dosage applied. PMID- 11728847 TI - Procedural learning improvements after six weeks of clozapine treatment. PMID- 11728848 TI - A randomized controlled pilot study of cognitive behavioral social skills training for older patients with schizophrenia. PMID- 11728849 TI - Psychological autopsy studies--a review. AB - Psychological autopsy is one of the most valuable tools of research on completed suicide. The method involves collecting all available information on the deceased via structured interviews of family members, relatives or friends as well as attending health care personnel. In addition, information is collected from available health care and psychiatric records, other documents, and forensic examination. Thus a psychological autopsy synthesizes the information from multiple informants and records. The early generation of psychological autopsies established that more than 90% of completed suicides have suffered from usually co-morbid mental disorders, most of them mood disorders and/or substance use disorders. Furthermore, they revealed the remarkable undertreatment of these mental disorders, often despite contact with psychiatric or other health care services. More recent psychological autopsy studies have mostly used case-control designs, thus having been better able to estimate the role of various risk factors for suicide. The future psychological autopsy studies may be more focused on interactions between risk factors or risk factor domains, focused on some specific suicide populations of major interest for suicide prevention, or combined psychological autopsy methodology with biological measurements. PMID- 11728850 TI - Relationships between attempted suicide, temperamental vulnerability, and violent criminality in a Swedish forensic psychiatric population. AB - The relationship between suicide attempts and different vulnerability factors was investigated in 61 male subjects during forensic psychiatric examination. Personality traits and psychopathy were determined by means of the Karolinska Scales of Personality (KSP) and the Psychopathy Checklist-Revised (PCL-R). Clinical diagnoses were determined by the use of Structured Clinical Interview for DSM-III-R (SCID). Platelet monoamine oxidase (MAO) activity was investigated in 58 subjects. Suicide attempters had extremely low socialization and high impulsive aggression. This was independent of a history of repeatedly criminal violence or not. They had higher PCL-R scores and lower levels of platelet MAO activity than non-attempters. No relationship was found between suicide attempts and repeated violent criminality. However, among the subjects with repeated criminal violence there was a highly significant difference between suicide attempters and non-attempters, indicating different personality profiles in violent offenders with and without suicidal behavior. Suicidal behavior was significantly associated with borderline personality disorder, but not with any other single DSM-III-R disorder. The results show a far more severe personality disturbance in suicide attempters than in other violent patients in this type of population. Suicidal behavior should therefore be evaluated and treated per se. PMID- 11728851 TI - The influence of comorbidity on the prevalence of suicidal behaviour. AB - The existence of mental disorders is almost constant in subjects who try to kill themselves. In addition, a majority of attempters have more than one diagnosis. This is especially true if Axis II or Axis I subthreshold conditions are taken into account. The existence of a disorder largely explains the association between most socioeconomic variables (sex, marriage, education level) and suicidality. Depressive disorders are the major risk factor, a risk probably linked to a current episode just before the attempt. The association to depressive episodes of an anxiety disorder or the existence of impulsive traits (and/or cluster B personality disorder or drug abuse) increases the risk of acting out. Ideation and attempts show parallel onset curves peaking between the ages of 14 and 20 years, with the existence of a previous DSM III R diagnosis as a strong predictor. The number of associated disorders linearly increases the probability of attempting suicide and is the only significant predictor for lethality. A proper treatment of mental disorders could substantially lower suicidality. PMID- 11728852 TI - Modelling suicide risk in affective disorders. AB - BACKGROUND: The lifetime risk of suicide in affective disorders is commonly quoted as 15%. This stems from hospital populations of affective disorders. AIMS: To model the lifetime prevalence of suicide using data on completed suicides from one English Health District and community-based rates of prevalence of affective disorders. METHODS: A secondary analysis of a primary data set based on 212 suicides in North Staffordshire was undertaken. The population rates of psychiatric morbidity were obtained from the National Comorbidity Survey. RESULTS: The model suggests a lifetime prevalence rate of suicide for any affective disorder at 2.4%, with a rate for those uncomplicated by substance abuse, personality disorder or non-affective psychosis at 2.4%, and a rate for uncomplicated cases who had no mental health service contact at 1.1%. CONCLUSIONS: Lifetime prevalence rates of suicide in subgroups of affective disorders may be lower than the traditional rates cited for hospital depression. This has implications for primary care projects designed to investigate the occurrence of and the prevention of suicide. PMID- 11728853 TI - Can better recognition and treatment of depression reduce suicide rates? A brief review. AB - Depression, the major cause of suicide, is prevalent but an under-detected, underdiagnosed and, under-treated illness and it is particularly true for depressed suicide victims. However, several studies consistently show that successful treatment of depression not only relieves depressive symptoms, but also decreases and makes suicidality vanish. If the rate of treated depressions in the population increases gradually, at a given point it will appear in the decline of the suicide rates. Although absolute evidence is lacking at present, recent reports from some European countries strongly suggest that increasing utilisation of antidepressants is one of the most important contributing factors in the decrease in suicide rates. PMID- 11728854 TI - Biological markers for suicidal behavior in alcohol dependence. AB - Alcohol-dependent populations have a high lifetime suicide rate (between 7 and 15%, relative risk = 7), and alcoholism is one of the two psychiatric disorders most frequently found in suicidal cases (between 15 and 25%). Biological factors that would detect patients at risk could thus be of value. Carbohydrate-deficient transferrin, monoamine oxidase B, soluble interleukin-2 receptor and cholesterol have been proposed as markers of suicidal risk in alcohol-dependent patients, although nonspecific and with low predictive value. On the other hand, there is large and convergent data stressing the importance of serotonin dysregulation as increasing the risk for aggressive behaviour toward the self, although it is not clear whether serotonin is involved through the altered behavior inhibition system, enhancement of anxiety and depression, or association with specific subtypes of alcohol-dependence, such as early-onset type II alcoholism. Considering the complex but significant impact of alcohol on serotonin metabolism and turnover, it is likely that serotonin mediates a large part of the proneness of ethanol to commit impulsive-aggressive behavior. PMID- 11728855 TI - The scientific and ethical basis for placebo-controlled trials in depression and schizophrenia: an FDA perspective. AB - There is a tension between the need for scientifically valid trials of new psychotropic drugs and concern about conducting placebo-controlled trials, the trials psychopharmacologists consider the gold standard trial, when this requires that some patients be denied existing effective therapy. This paper will review the scientific principles supporting the need for placebo-controlled trials in depression and schizophrenia, and will provide preliminary data on failure rates of placebo-controlled trials for these disorders, as illustrations of the application of these principles. Next, the ethical issues pertinent to the conduct of placebo-controlled trials for these two serious psychiatric disorders will be reviewed. Preliminary data on suicides in placebo-controlled depression and schizophrenia trials will be presented to argue for the ethical acceptability of the conduct of placebo-controlled trials in these two conditions. PMID- 11728856 TI - Role of dopamine in non-depressed patients with a history of suicide attempts. AB - Several data are available about the implication of the dopaminergic system in the control of inward-directed aggression. Previously, we suggested an involvement of D2-dopaminergic function in the expression of suicidal behavior by demonstrating a smaller growth hormone (GH) response to apomorphine, a dopaminergic agonist, in depressed patients with a history of suicide attempts in comparison to non-attempters. In the present study, in order to test this hypothesis, GH responses to intravenous apomorphine were measured in non depressed patients with a history of suicide attempts. The study was performed in 17 non-depressed male patients with a score less than 12 on the 17-item HAMD. The patients were subgrouped into suicide attempters (N = 7) and non-attempters (N = 10). Mean GH peak responses to apomorphine differed significantly between suicide attempters and non-attempters: (mean +/- SD) for GH peak, 10.4 +/- 8.2 ng/mL vs 27.3 +/- 13.1 ng/ml, F = 9.0, P = 0.009. In conclusion, dopaminergic disturbances seem to play a role in the biology of inward-directed aggression in non-depressed patients. PMID- 11728857 TI - Dexamethasone suppression test and suicide attempts in schizophrenic patients. AB - The suicide attempts were assessed in 32 schizophrenic patients on whom the dexamethasone suppression test (DST) was done twice in the course of illness: in the years 1985-91 and 1996-97. In the 1985-91 period, both baseline and post dexamethasone cortisol levels were significantly higher in the patients with previous suicide attempts and baseline cortisol was higher in the patients who were to make a future attempt. In 1996-97, DST non-suppression was shown in more than half of the patients with a history of suicide attempt and in none of those without such history: all cortisol levels were significantly higher in the patients with a history of suicide attempt. Although the mean intensity of depression was higher in the patients with a history of suicide attempt, no association between the intensity of depression and present or previous DST non suppression status was found. It is suggested that the hyperactivity of the hypothalamic-pituitary-adrenal axis may constitute an element of diathesis for suicidal behavior in schizophrenic patients. PMID- 11728858 TI - Suicide in military circles in Italy (1986-1998). PMID- 11728859 TI - Collective efforts to modulate the host actin cytoskeleton by Salmonella type III secreted effector proteins. PMID- 11728861 TI - Are the structures of SarA and SarR similar? PMID- 11728862 TI - Polymorphic proteins of Chlamydia spp.--autotransporters beyond the Proteobacteria. AB - Gram-negative bacteria secrete a variety of proteins to the cell surface and beyond, a process with many inherent difficulties. An exceptionally widespread answer to these problems is the type V (or autotransporter) secretion pathway. By exploiting the data made available by bacterial genome sequencing, we have discovered that the previously described polymorphic proteins of Chlamydia spp. resemble members of the autotransporter family, and we suggest that they follow the same secretion pathway. PMID- 11728863 TI - Microbial genomics. Dynamic duos. PMID- 11728864 TI - Lifestyles of the intracellular. PMID- 11728866 TI - Putting C. elegans to Gram-positive use. PMID- 11728869 TI - Shiga toxins--not just cytotoxins anymore. PMID- 11728867 TI - Supermolecular structure of the EPEC TTSS. PMID- 11728870 TI - Symbiosis and pathogenesis: common themes, different outcomes. PMID- 11728871 TI - The role of bacterial pili in protein and DNA translocation. AB - Gram-negative bacteria have surface appendages that assemble via different secretion machineries. Recently, new experimental approaches have contributed to a better understanding of the molecular mechanisms of flagellar and pilus assembly, and protein secretion. These findings can be applied to plant pathogenic bacteria, which probably transfer effector proteins directly into their eukaryotic host cells. Here, it is suggested that assembly of Hrp pili occurs in the periplasm and that unfolded effector proteins attach to pilins within the pili, thus effecting protein translocation. A two-domain structure for the HrpA pilin from Pseudomonas syringae is also predicted. PMID- 11728872 TI - Virulence in Candida species. AB - Candida species other than Candida albicans now account for up to 50% of deep candidiasis cases, yet little attention has been paid to the virulence attributes of these fungi. Adherence to host tissues, response to environmental changes and the secretion of hydrolases are all thought to be important in Candida virulence. The identification of virulence attributes unique to a particular Candida species could provide powerful insights into the pathogenic process but will require the use of genome-wide approaches such as transcript profiling, signature-tagged mutagenesis and in vivo expression technology. PMID- 11728873 TI - Mycobacterial persistence: adaptation to a changing environment. AB - Mycobacterium tuberculosis is a bacterial pathogen that can persist within an infected individual for extended periods of time without causing overt, clinical disease, in a state normally referred to as latent or chronic tuberculosis. Although the replicative state of the bacterium during this period is a matter of some conjecture, recent developments have indicated that the bacterium requires the regulated expression of a set of genes and metabolic pathways to maintain a persistent infection in an immunocompetent host. The characterization of these gene products and their role in bacterial metabolism and physiology is starting to provide insights into the mechanisms that M. tuberculosis has evolved to adopt its highly successful mode of pathogenicity. PMID- 11728874 TI - What determines nasal carriage of Staphylococcus aureus? AB - Nasal carriage of Staphylococcus aureus is an important risk factor for infection by this organism in both community and hospital settings; this article reviews the role of host and bacterial factors in carriage. A host genetic influence appears likely but the phenotypic determinants are unknown. Possibilities include variability in host adhesins, immune response or secretion of antimicrobial molecules. Colonization resistance by S. aureus, together with the observation that persistent carriers often carry a single strain whereas intermittent carriers can be colonized with unrelated strains over time, suggests that bacterial factors could also be involved. PMID- 11728875 TI - Exploiting genomics to discover new antibiotics. AB - There is an urgent need to develop new classes of antibiotics to tackle the increase in resistance in many common bacterial pathogens. One strategy to develop new antibiotics is to identify and exploit new molecular targets and this strategy is being driven by the wealth of new genome sequence information now available. Additionally, new technologies have been developed to validate new antibacterial targets, for example, new technologies have been developed to enable rapid determination of whether a gene is essential and to assess the transcription status of a putative target during infection. As a result, many novel validated targets have now been identified and for some, appropriate high throughput screens against diverse compound collections have been carried out. Novel antibiotic leads are emerging from these genomics-derived targeted screens and the challenge now is to optimize and develop these leads to become part of the next generation of antibiotics. PMID- 11728877 TI - Therapeutic vasculogenesis using human cord blood-derived endothelial progenitors. AB - Peripheral blood of adult species contains endothelial progenitor cells (EPCs) that participate in neovascularization, consistent with postnatal vasculogenesis. Abundant EPCs can be isolated from a relatively small volume of human umbilical cord blood, and that culture-expanded EPCs participate in endothelial network formation in vitro. Transplanted EPCs incorporated into sites of active neovascularization and formed capillaries among preserved skeletal myocytes in the ischemic hindlimb of athymic nude rats in vivo. Furthermore, transplantation of EPC quantitatively and effectively augmented neovascularization in response to hindlimb ischemia. Thus, human umbilical cord blood seems to be a novel source for EPCs, and the transplantation of cord blood-derived EPCs may become a useful strategy to modulate postnatal neovascularization. PMID- 11728878 TI - Tissue factor in the heart. Multiple roles in hemostasis, thrombosis, and inflammation. AB - Tissue factor (TF) is the primary cellular initiator of the coagulation protease cascade. It plays an essential role in hemostasis to limit hemorrhage in the event of vascular injury. Indeed, loss of TF is incompatible with life. Recent studies have suggested that TF also plays non-hemostatic roles in blood vessel development, tumor angiogenesis, tumor metastasis, cell migration, and inflammation. This review discusses the roles of TF in hemostasis, thrombosis, and inflammation in the heart as well as other potential roles of TF in the maintenance of cardiac muscle. PMID- 11728879 TI - Cloning of aged animals: a medical model for tissue and organ regeneration. AB - Cloning by nuclear transfer has great potential application in pharmaceutical protein production, xeno-transplantation, and perhaps most excitingly, therapeutic cloning. In therapeutic cloning a patient's own skin cells can be used to generate cloned embryos from which embryonic stem cells are isolated. Through targeted differentiation, embryonic stem cells can be directed to develop into the desired tissues/organs for replacement. The combination of homologous recombination of genes and nuclear transfer also offers the promise of correcting defective genes in humans. Demonstration of the successful cloning of aged animals is important for these future medical applications because degenerative diseases often afflict older adults. Our studies have demonstrated that skin fibroblast cells from aged adults, even after prolonged culture, provide nuclear donors equally as competent for cloning as cells from young adults or fetuses. These findings have paved the way for medically treating degenerative diseases of aged humans by tissue regeneration technologies made possible through cloning and homologous recombination. PMID- 11728880 TI - SOX18 and the transcriptional regulation of blood vessel development. AB - SOX18 is a transcription factor that is transiently expressed in nascent endothelial cells during embryonic development and adult neovascularization. This protein belongs to the SOX family of transcription factors, which are proving to be some of the key regulators of cell-type specification in the vertebrate embryo. Natural mutations in the Sox18 gene have been shown to result in cardiovascular dysfunction, in some cases leading to death. Available evidence thus implicates Sox18 as an important regulator of vascular development, most likely playing a key role in endothelial cell specification. However, the genetic knockout of Sox18 in mice has produced a confounding result that complicates our understanding of the molecular mode of action of the SOX18 protein. We speculate that Sox18 may act in a redundant fashion with closely related genes such as Sox7 and/or Sox17. PMID- 11728881 TI - Dual effects of angiotensin II type 2 receptor on cardiovascular hypertrophy. AB - Roles of angiotensin II (Ang II) in the regulation of cardiovascular system under normal and pathological condition have been well documented. Although two major subtypes of Ang II receptors, AT(1) and AT(2), are found in various proportions, the role and signaling mechanisms of AT(2) in the control of hypertrophic responses of cardiac ventricle and vasculature are not clear. Although earlier reports indicated that AT(2)'s functions are essentially growth suppression, an increasing number of recent reports indicate that AT(2) in cardiovascular tissues are often growth promoting. In some tissues AT(1) and AT(2) seem to share a common signaling pathways, at least in part. This review focuses on the accumulating evidence for the AT(2) function in the cardiovascular system. PMID- 11728882 TI - A novel role for serotonin in heart. AB - Congenital heart disease is a major cause of disability and morbidity, often initiated by both environmental components and genetic susceptibility. Identification of factors controlling myocardial differentiation and proliferation is of great importance for understanding the pathogenesis of congenital heart diseases. Several lines of evidence suggest that serotonin [5 hydroxytryptamine (5-HT)] regulates cardiovascular functions during embryogenesis and adulthood. However, the molecular mechanism by which 5-HT regulates embryonic development of heart and cardiovascular functions remained unknown until recently. Inactivation of the 5-HT(2B) receptor (5-HT(2B)R) gene leads to embryonic and neonatal death due to the following defects in the heart: (a) 5 HT(2B)R mutant embryos exhibit a lack of trabeculae in the heart and a reduction in the expression levels of a tyrosine kinase receptor, called ErbB-2, leading to mid-gestation lethality. These in vivo data suggest that the Gq-coupled 5-HT(2B)R uses the signaling pathway of the tyrosine kinase receptor ErbB-2 for cardiac differentiation. (b) Newborn 5-HT(2B)R mutant mice exhibit cardiac dilation resulting from contractility deficits and structural deficits at the intercellular junctions between cardiomyocytes. (c) In adult 5-HT(2B)R mutant mice, echocardiography and electrocardiography confirm the presence of left ventricular dilation and decreased systolic function. These results constitute the first genetic evidence that 5-HT via the 5-HT(2B)R, regulates differentiation and proliferation during development as well as cardiac structure and function in adults. PMID- 11728883 TI - Cell biology of tissue factor, an unusual member of the cytokine receptor family. AB - Tissue factor (TF) is a 47-kDa transmembrane protein, involved in the onset of coagulation. However, it also influences pathophysiological processes such as inflammation and tumor angiogenesis. Although the molecular mechanisms responsible for these phenomena remain unclear, it is thought that they are brought about by the action of intracellular signaling, resulting in gene transcription and subsequent protein synthesis. In this review we focus on FVIIa/TF-induced intracellular signaling and its possible role in physiology. PMID- 11728884 TI - Salicylates and vascular smooth muscle cell proliferation: molecular mechanisms for cell cycle arrest. AB - Salicylates are effective prophylactic treatment strategies for myocardial infarction and ischemic strokes. Recent evidence suggests that high doses of salicylates may exert direct, platelet-independent effects on the vascular wall. Salicylate and aspirin, in concentrations between 1 and 5 mM, effectively inhibit vascular smooth muscle cell proliferation and DNA synthesis without inducing cellular toxicity or apoptosis. This inhibition is associated with effects on specific cell-cycle regulatory molecules, and may proceed via downregulation of the transcription factor, nuclear factor (NF)-kappaB. High-dose salicylates and selective NF-kappaB inhibitors may, therefore, play an important role in the management of vascular proliferative disorders. PMID- 11728886 TI - The health profile of Spanish incarcerated delinquent youths. PMID- 11728887 TI - Risk factors for eating disorders in Italian adolescents. PMID- 11728888 TI - The relative influence of smoking on drinking and drinking on smoking among high school students in a rural tobacco-growing county. AB - PURPOSE: To construct and estimate a model that contains a reciprocal relationship between smoking and drinking and to test whether the gateway drug thesis or the cumulative risk behaviors thesis best fits the data. METHODS: Data (n = 630) are from a survey of all students (50% female; aged 16.2 years on average; 2.4 grade point average; and 57% residing in homes with both mother and father present) in a rural, tobacco-growing county's two high schools, one public (85%) and one private, in 1993. The survey was conducted by the schools as part of their alcohol and other drugs (AOD) prevention programs and was coordinated by the county AOD Council. Students completed the questionnaires in their homerooms. Endogenous predictors of drinking and smoking include student's perception of adult drug behavior, peer pressure to drink, degree to which their friends' drink, and attitudes toward drinking and smoking. Path coefficients were estimated by using LISREL. RESULTS: The strong correlation between smoking and drinking resulted from shared causes, rather than from the effects of one type of drug use on the other. Approval of drinking had the strongest association with being a drinker (beta =.57) and with being a smoker (beta =.37). Those who found smoking offensive were less likely to be a smoker (beta = -.25). However, attitude toward smoking was not associated with being a drinker. Having drinking friends increased both the likelihood of being a drinker (beta =.29) and of being a smoker (beta =.23). Peer pressure to drink increased the likelihood of being a smoker (beta =.14) and of being a drinker (beta =.12). Students with lower grade point averages, males, older students, students in public school, and students with family structures other than both parents living in the same household were more likely to be a drinker and were more likely to be a smoker. CONCLUSIONS: Our results support the cumulative risk behaviors thesis. The link between both high risk behaviors, smoking and drinking, results from common causes rather than from drinking leading to smoking. PMID- 11728889 TI - Factors associated with use of preventive dental and health services among U.S. adolescents. AB - PURPOSE: To examine adolescents' use of preventive medical and dental services and its relationship to demographic characteristics and other variables reflecting access to and need for care. METHODS: Self- and parent-reported data from a sample of 5644 adolescents aged 11 to 21 years from the National Longitudinal Study of Adolescent Health (Add Health). Variables studied include the influence of both the adolescents' demographic and socioeconomic characteristics (age, race/ethnicity, place of birth, acculturation, insurance status, and perception of health), as well as those of their parents (race/ethnicity, income, level of education, place of birth) on their lifetime use and use within the past year of medical and dental services. Bivariate and logistic regression analyses were conducted using SAS and SUDAAN. RESULTS: Approximately 32% of respondents had not had a physical examination in the year before the survey, and the same percentage had not had a dental examination. Approximately 2% reported never having had either a physical or a dental examination. Logistic regression reveals that lack of insurance, low family income, and low parental education level are significantly associated with the lack of preventive medical care. Lack of an annual dental visit was associated with male gender; black, Hispanic, or mixed race/ethnicity; and lack of insurance. Never having had a dental visit was the only dependent variable found to be associated with place of birth. CONCLUSIONS: Health insurance and family income are most consistently related to adolescents' use of preventive medical and dental care. However, the relationship between lack of dental care and place of birth emphasizes the need to improve access to dental services for immigrant teens. These findings are particularly relevant as states design systems of care for adolescents under the State Children's Health Insurance Program. PMID- 11728890 TI - Characteristics of vegetarian adolescents in a multiethnic urban population. AB - PURPOSE: To examine the prevalence of adolescents' vegetarianism in a multiethnic, urban population, and its correlates with demographic, personal, weight-related, and behavioral factors. METHODS: Self-report and anthropometric data were collected from a representative sample of 4746 adolescents from 31 public middle schools and high schools in the Twin Cities area of Minnesota. Students answered questions concerning vegetarianism, food and weight, and health behaviors. Height and weight were directly measured. Comparisons were made between self-reported vegetarians and nonvegetarians; these analyses also assessed gender and race/ethnicity interactions. In the second set of analyses, demographic and behavioral characteristics of more restricted and semi vegetarians were examined. Analyses were done by logistic regression. RESULTS: Teenage vegetarians comprise about 6% of the sample. The vegetarians were more likely than nonvegetarians to be female, not black, weight- and body-conscious, dissatisfied with their bodies, and involved in a variety of healthy and unhealthy weight control behaviors. Vegetarians more often reported having been told by a physician that they had an eating disorder and were more likely to have contemplated and attempted suicide. Vegetarian males were found to be an especially high risk group for unhealthy weight control practices. Few ethnic group differences among vegetarians were noted. Adolescents who did not eat chicken and fish were at lower risk than those who also ate chicken and fish. CONCLUSIONS: Adolescent vegetarians are at greater risk than others for involvement in unhealthy and extreme weight control behaviors. Vegetarian males are at particularly high risk. Vegetarianism among adolescents may therefore be a signal for preventive intervention. Adolescents who choose to become vegetarians may also need to learn how to healthfully do so. PMID- 11728891 TI - Effects of a brief, theory-based STD-prevention program for female college students. AB - PURPOSE: To evaluate a brief, theoretically guided sexually transmitted disease (STD) risk-reduction intervention tailored to college-aged women. METHODS: The participants were 78 undergraduate females (M = 20 years; 76% European-American) who reported inconsistent condom use or multiple sexual partners. Participants were randomly assigned to one of three groups: (a) a one-session intervention based on the information-motivation-behavioral skills (IMB) model, (b) a one session information-only intervention (INFO), or (c) a wait-list control (WLC) group. Consistent with Fisher and Fisher's (1992) IMB model, we predicted that risk reduction would be greater when information about HIV was supplemented with motivational enhancement strategies and skills training. To evaluate this hypothesis, groups were compared at the post-intervention assessment and at a 2 month follow-up using analyses of covariance and log odds ratios. RESULTS: At the post-intervention assessment, the IMB and INFO groups demonstrated increased STD related knowledge. At 2-month follow-up, the IMB and INFO groups showed sustained STD-related knowledge, and the IMB group showed reductions in number of sexual partners compared to the WLC group. CONCLUSIONS: These results provide partial support for the hypothesis that an IMB model-based intervention leads to reductions in sexual risk behavior and suggest directions for future research. PMID- 11728892 TI - Quality of life and health-risk behaviors among adolescents. AB - PURPOSE: To assess the association between health-risk behaviors and self perceived quality of life among adolescents METHODS: A sample of 2801 students (957 seventh and eighth graders and 1844 ninth through twelfth graders) completed the Teen Assessment Survey (TAP) and the surveillance module of the Youth Quality of Life Instrument (YQOL-S). TAP responses were used to determine health-risks related to tobacco use, alcohol use, illicit drug use, and high risk sexual behavior. Separate multivariate analyses of variance showed mean differences in contextual and perceptual items of the YQOL-S for each health-risk behavior. Differences among engagers (adolescents who often engage), experimenters (occasionally engage), and abstainers (never engage) in the health-risk behavior were evaluated by gender and junior/senior high school groups. RESULTS: In general, adolescent abstainers reported higher quality of life (QoL) than engagers and experimenters on YQOL-S items. Adolescents who engaged in multiple risk behaviors scored even lower than those who engaged in only one health-risk behavior. Experimenters tended to rate their QoL more similar to that of abstainers than to that of engagers. CONCLUSIONS: The framework of QoL proved useful in the evaluation of adolescents' engagement in health-risk behaviors. Additionally, assessing the areas of QoL that differ between the groups may provide information for planning interventions aimed at risk reduction among engagers and experimenters. PMID- 11728894 TI - School-based health center: position paper of the Society for Adolescent Medicine. PMID- 11728893 TI - West African Youth Initiative: outcome of a reproductive health education program. AB - PURPOSE: To describe the implementation and evaluation of an adolescent reproductive health peer education program in West Africa. The program, known as the West African Youth Initiative (WAYI), was developed to improve knowledge of sexuality and reproductive health, and promote safer sex behaviors and contraceptive use among sexually active adolescents in Nigeria and Ghana. METHODS: Between November 1994 and April 1997, two organizations, the Association for Reproductive and Family Health (ARFH), based in Nigeria, and Advocates for Youth, based in Washington D.C., supported community-based youth-serving organizations in the two countries to implement peer education projects. Consultants from the African Regional Health Education Centre (ARHEC) in Nigeria provided technical assistance in designing and conducting a quasi-experimental process and outcome evaluation of the projects. RESULTS: There were significant differences over time and between intervention and control groups concerning reproductive health knowledge, use of contraceptives in the previous 3 months, willingness to buy contraceptives, and self-efficacy in contraceptive use. CONCLUSIONS: Overall, the project provides evidence that peer education is most effective at improving knowledge and promoting attitudinal and behavior change among young people in school settings. PMID- 11728895 TI - Implicit learning revealed by the method of opposition. AB - Complex information, such as that required for motor skills, can be learned implicitly, without awareness. Much debate has centered on the appropriate methods for proving that implicit learning is not influenced by explicit awareness. A recent study by Destrebecqz and Cleeremans has provided compelling evidence for implicit sequence learning without awareness by using the 'method of opposition'. PMID- 11728896 TI - 'Action binding': dynamic interactions between vision and touch. PMID- 11728897 TI - Thinking Beyond the Fringe. PMID- 11728898 TI - Vision scientists go (far) east. PMID- 11728899 TI - I don't know the name, but the facade rings a bellellipsis. PMID- 11728901 TI - Predicting the unpredictable. PMID- 11728900 TI - Acquiring languages - two for the price of one? PMID- 11728902 TI - Smiling eyes. PMID- 11728903 TI - Priming: a tool for imaging. PMID- 11728904 TI - Hand-waving insight into infant language. PMID- 11728906 TI - The colour of smell. PMID- 11728905 TI - Robots help humans defeat robots. PMID- 11728907 TI - Evolution in the sky. PMID- 11728908 TI - Word acquisition reflects lateralization of hand skill. AB - Right hand preference distinguishes Homo sapiens from our closest primate relative, the chimpanzee. Are differences in degrees of handedness associated with differences in the rate of language development? To answer this question, verbal performance is examined in relation to hand skill in a UK national birth cohort dataset. Using 3-D data plots, we show that increasing dominant-hand skill is associated with increasing verbal ability, and stronger lateralization is associated with earlier acquisition of words. Thus, right-handed bias is relevant to the lateralization of language; variation along this dimension may represent species-specific genetic or 'epigenetic' diversity. PMID- 11728909 TI - Brain reflections of words and their meaning. AB - The neurobiological organization of meaningful language units, morphemes and words, has been illuminated by recent metabolic and neurophysiological imaging studies. When humans process words from different categories, sets of cortical areas become active differentially. The meaning of a word, more precisely aspects of its reference, may be crucial for determining which set of cortical areas is involved in its processing. Word-related neuron webs with specific cortical distributions might underlie the observed category-specific differences in brain activity. Neuroscientific principles can explain these differential topographies. PMID- 11728910 TI - Impaired processing of rapid stimulus sequences in dyslexia. AB - Apart from their reading difficulties, dyslexic subjects often suffer from a variety of subtle sensory and motor deficits. Whether these deficits have a causal relationship to the reading disorder, form additional risk factors, or are totally independent of the reading problem, is under vivid debate. In this article, we review the evidence and suggest that 'sluggish attentional shifting' (SAS) can account for the impaired processing of rapid stimulus sequences in dyslexia. Within this novel framework attention-related prolongation of input chunks is decisive for many small deficits found in dyslexic subjects. PMID- 11728911 TI - Episodic future thinking. AB - Thinking about the future is an integral component of human cognition - one that has been claimed to distinguish us from other species. Building on the construct of episodic memory, we introduce the concept of 'episodic future thinking': a projection of the self into the future to pre-experience an event. We argue that episodic future thinking has explanatory value when considering recent work in many areas of psychology: cognitive, social and personality, developmental, clinical and neuropsychology. Episodic future thinking can serve as a unifying concept, connecting aspects of diverse research findings and identifying key questions requiring further reflection and study. PMID- 11728912 TI - Sequential learning in non-human primates. AB - Sequential learning plays a role in a variety of common tasks, such as human language processing, animal communication, and the learning of action sequences. In this article, we investigate sequential learning in non-human primates from a comparative perspective, focusing on three areas: the learning of arbitrary, fixed sequences; statistical learning; and the learning of hierarchical structure. Although primates exhibit many similarities to humans in their performance on sequence learning tasks, there are also important differences. Crucially, non-human primates appear to be limited in their ability to learn and represent the hierarchical structure of sequences. We consider the evolutionary implications of these differences and suggest that limitations in sequential learning may help explain why non-human primates lack human-like language. PMID- 11728913 TI - Where intention is born. PMID- 11728914 TI - Building bridges. PMID- 11728915 TI - Triumph or travesty? PMID- 11728916 TI - Triumph of Sociobiology. PMID- 11728917 TI - Statins and their potential for osteoporosis. PMID- 11728918 TI - Molecular mechanisms of glucocorticoid-induced osteoporosis. AB - Bone loss resulting from long-term glucocorticoid therapy is common and clinically relevant. A number of different glucocorticoid-mediated effects are responsible for the reduction in bone density: (i) glucocorticoid-induced direct impairment of osteoblast, osteocyte, and osteoclast function leads to reduced bone remodeling and diminished repair of microdamage in bone; (ii) the effects of parathyroid hormone (PTH) might be more pronounced in the presence of glucocorticoids, whereas vitamin D plays a lesser role in the pathogenesis of steroid-induced osteoporosis; (iii) glucocorticoids antagonize gonadal function and inhibit the osteoanabolic action of sex steroids; and (iv) increased renal elimination and reduced intestinal absorption of calcium lead to a negative calcium balance that has been suggested to promote secondary hyperparathyroidism. From a mechanistic point of view, all of the aforementioned effects have long been considered to be mediated at the molecular level exclusively by genomic actions. However, there is now increasing evidence for the existence of rapid glucocorticoid effects that are incompatible with this classical mode of action. These rapid effects, termed nongenomic effects, are mediated by glucocorticoid interactions with biological membranes, either through binding to membrane receptors or by physicochemical interactions. It is possible, but has yet to be shown, that these effects play a role in the pathogenesis of glucocorticoid induced osteoporosis. PMID- 11728919 TI - Alendronate for the prevention of bone loss in patients on inhaled steroid therapy. AB - One hundred women on inhaled steroid therapy (dose range from 800 to <1600 microg per day) were randomized to receive 10 mg of oral alendronate or placebo (with 500 mg of calcium in the form of daily calcium carbonate). Bone mineral density (BMD) was measured at baseline, 6 months, and 12 months. The percentage changes in BMD were -0.80% in the placebo group and 2.99% in the alendronate group at the spine (p < 0.001 by analysis of covariance [ANCOVA]), and were -0.51% in the placebo group and 0.97% in the alendronate group at the femoral neck (p < 0.05 by ANCOVA). Five patients in the alendronate-treated group, and a similar number of patients in the placebo group, complained of mild gastric discomfort. We conclude that women on inhaled steroid therapy were at risk of accelerated bone loss, which could be prevented by a daily dose of 10 mg of alendronate. PMID- 11728920 TI - A theoretical analysis of the contributions of remodeling space, mineralization, and bone balance to changes in bone mineral density during alendronate treatment. AB - In patients with osteoporosis, alendronate treatment causes an increase in bone mineral density (BMD) and a decrease in fracture incidence. Alendronate acts by changing the bone remodeling process. Changes in bone remodeling resulting in decreased remodeling space, increased bone balance per remodeling cycle, and increased mineralization (ash mass/bone mass) have all been associated with alendronate treatment. Understanding the relative contributions of these parameters to BMD increases could help predict the utility of long-term (>10 years) or intermittent treatment strategies, as well as treatment strategies in which another pharmaceutical is administered concurrently. We have developed a computer simulation of bone remodeling to compare the contributions of focal bone balance and mineralization on BMD by simulating alendronate treatment using a bone balance method (decreased remodeling space, increased focal bone balance, uniform bone mineralization) and a mineralization method (decreased remodeling space, neutral focal bone balance, varying bone mineralization). Although both methods are able to predict BMD increases caused by alendronate over short periods, our findings suggest that the mineralization method may be more descriptive of long-term alendronate treatment. This implies that mineralization may be a larger contributor to BMD changes caused by alendronate than the focal bone balance. Based on this finding we offer a hypothesis to describe how remodeling space, focal bone balance, and mineralization each contribute to alendronate-induced BMD changes. Future analyses with this method could be used to identify improved dosing regimens and to predict which osteoporosis treatments would best complement each other. PMID- 11728921 TI - Epidemiology of fractures in England and Wales. AB - Records from the General Practice Research Database were used to derive age- and gender-specific fracture incidence rates for England and Wales during the period 1988-1998. In total, 103,052 men and 119,317 women in the sample of 5 million adults sustained a fracture over 10.4 million and 11.2 million person-years (py) of follow-up. Among women, the most frequent fracture sites were the radius/ulna (30.2 cases per 10,000 py) and femur/hip (17.0 per 10,000 py). In men, the most common fracture was that of the carpal bones (26.2 per 10,000 py); the incidence of femur/hip fracture was 5.3 per 10,000 py. Varying patterns of fracture incidence were observed with increasing age; whereas some fractures became more common in later life (vertebral, distal forearm, hip, proximal humerus, rib, clavicle, pelvis), others were more frequent in childhood and young adulthood (tibia, fibula, carpus, foot, ankle). The lifetime risk of any fracture was 53.2% at age 50 years among women, and 20.7% at the same age among men. Whereas fractures of the proximal femur and vertebral body were associated with excess mortality over a 5 year period following fracture diagnosis among both men and women, fractures of the distal forearm were associated with only slight excess mortality in men. This study provides robust estimates of fracture incidence that will assist health-care planning and delivery. PMID- 11728922 TI - Human osteoprogenitor growth and differentiation on synthetic biodegradable structures after surface modification. AB - The ability to generate new bone for skeletal use is a major clinical need. Biomimetic scaffolds that interact and promote osteoblast differentiation and osteogenesis offer a promising approach to the generation of skeletal tissue to resolve this major health-care issue. In this study we examine the ability of surface-modified poly(lactic acid) (PLA) films and poly(lactic-co-/glycolic acid) (PLGA) (75:25) porous structures to promote human osteoprogenitor adhesion, spreading, growth, and differentiation. Cell spreading and adhesion were examined using Cell Tracker green fluorescence and confocal microscopy. Osteogenic differentiation was confirmed with alkaline phosphatase activity as well as immunocytochemistry for type I collagen, core binding factor-1 (Cbfa-1), and osteocalcin. Poor cell growth was observed on nonmodified PLA films and PLGA scaffolds. The polymers were then coupled with RGD peptides [using poly(L lysine), or PLL] and physical adsorption as well as PLA films presenting adsorbed fibronectin (FN). Both modifications enhanced cell attachment and spreading. On PLA-FN and PLA-PLL-GRGDS films, the osteoblast response was dose dependent (20 pmol/L to 0.2 micromol/L FN and 30 nmol/L to 30 micromol/L PLL-GRGDS) and significant at concentrations as low as 2 nmol/L FN and 30 nmol/L PLL-GRGDS. With optimal concentrations of FN or RGD, adhesion and cell spreading were comparable to tissue culture plastic serum controls. In PLGA (75:25) biodegradable porous scaffolds, coated with FN, PLL-GRGDS, or fetal calf serum for 24 h in alpha MEM alone, prior to growth in dexamethasone and ascorbate-2-phosphate for 4-6 weeks, extensive osteoblast impregnation was observed by confocal and fluorescence microscopy. Cell viability in extended culture was maintained as analyzed by expression of Cell Tracker green and negligible ethidium homodimer-1 (a marker of cell necrosis) staining. Alkaline phosphatase activity, type I collagen, Cbfa-1, and osteocalcin expression were observed by immunocytochemistry. Mineralization of collagenous matrix took place after 4 weeks, which confirmed the expression of the mature osteogenic phenotype. These observations demonstrate successful adhesion and growth of human osteoprogenitors on protein- and peptide-coupled polymer films as well as migration, expansion, and differentiation on three dimensional biodegradable PLGA scaffolds. The use of peptides/proteins and three dimensional structures that provide positional and environmental information indicate the potential for biomimetic structures coupled with appropriate factors in the development of protocols for de novo bone formation. PMID- 11728923 TI - Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. AB - We compared the gene expression profiles of human dental pulp stem cells (DPSCs) and bone marrow stromal stem cells (BMSSCs) as representative populations of odontoprogenitor and osteoprogenitor cells, respectively. Total RNA from primary cultures was reverse-transcribed to generate cDNA probes and then hybridized with the Research Genetics human gene microarray filter GF211. The microarrays were analyzed using the PATHWAYS software package. Human DPSCs and BMSSCs were found to have a similar level of gene expression for more than 4000 known human genes. A few differentially expressed genes, including collagen type XVIII alpha1, insulin-like growth factor-2 (IGF-2), discordin domain tyrosine kinase 2, NAD(P)H menadione oxidoreductase, homolog 2 of Drosophila large disk, and cyclin dependent kinase 6 were highly expressed in DPSCs, whereas insulin-like growth factor binding protein-7 (IGFBP-7), and collagen type I alpha2 were more highly expressed in BMSSCs. Furthermore, we confirmed the differential expression of these genes by semiquantitative polymerase chain reaction (PCR) and northern blot hybridization. The protein expression patterns for both IGF-2 and IGFBP-7 correlated with the differential mRNA levels seen between DPSCs and BMSSCs. This report describes the gene expression patterns of two distinct precursor populations associated with mineralized tissue, and provides a basis for further characterization of the functional roles for many of these genes in the development of dentin and bone. PMID- 11728924 TI - Effects of triiodothyronine on the insulin-like growth factor system in primary human osteoblastic cells in vitro. AB - Thyroid hormone plays a major role in the regulation of bone metabolism but the mechanism by which this is accomplished is not clear. Interactions of thyroid hormone with the growth hormone/insulin-like growth factors (IGFs) axis suggest an alternate pathway of action for triiodothyronine (T(3)) on bone formation, besides direct effects. The present study investigates the influence of T(3) on IGF-1, IGF-2, IGF-1 receptor (IGF-1R), and IGF binding protein (IGFBP) transcripts, and on IGF-1 action in human osteoblastic cells (hOB) under serum free culture conditions. No influence of T(3) on IGF-1, IGF-2, IGFBP-3, or IGFBP 4 mRNA levels in hOB was observed. However, T(3) at concentrations of 10(-8) mol/L and 10(-7) mol/L increased IGF-1R mRNA levels in a dose-dependent manner (p < 0.01) and enhanced IGFBP-5 mRNA levels at a concentration of 10(-7) mol/L (p < 0.05), as assessed by reverse transcriptase-polymerase chain reaction. Correspondingly, Scatchard analysis of [(125)I]-IGF-1 binding revealed that T(3) at 10(-7) mol/L increased the number of IGF-1 binding sites in hOB, with small changes in receptor affinity. In addition, a synergistic effect of T(3) and IGF-1 on hOB proliferation was found (p < 0.05). We conclude that IGF-1R and IGFBP-5 are thyroid hormone target genes in human osteoblasts, whereas IGF-1 mRNA expression itself appears not to be regulated by T(3) in hOB. However, T(3) stimulates IGF-1R mRNA expression as well as IGF-1 binding and IGF-1 induced cell proliferation in osteoblasts, thus suggesting thyroid hormone may potentiate the effect of IGF-1 at the receptor level. This may contribute to the positive effects of thyroid hormone on bone formation, which, in addition, may be modulated by increased IGFBP-5 expression. PMID- 11728925 TI - Fourier transform infrared microspectroscopic investigation of the maturation of nonstoichiometric apatites in mineralized tissues: a horse dentin study. AB - Fourier transform infrared microspectroscopy (FTIRM) was used to study carbonated apatite/collagen interactions and maturation in horse secondary dentin. Unlike human dentin, this model contains no peritubular material around the odontoblastic processes and is thus quite similar to bone in composition, but not subject to tissue turnover. Crystals close to the mineralization front were very immature, showing high HPO(4) and very low CO(3) levels. Carbonate ions were located essentially in very labile, reactive environments, probably on the crystal surface. Removal of some of the HPO(4) ions from crystals during maturation was linked to an increase in total carbonate content. The CO(3) ions in labile environments decreased, probably after incorporation into more organized regions of the lattice. However, this increase of total carbonate content was associated with greater mineral crystallinity, confirming findings in other studies of synthetic apatite maturation in vitro. The good correlation between these results and those of in vitro experiments suggests that crystal maturation is essentially due to physicochemical processes and that the organic matrix controls only crystal size, multiplication, and/or organization. PMID- 11728926 TI - Inhibition of bone resorption by alendronate and risedronate does not require osteoclast apoptosis. AB - Bisphosphonate inhibition of bone resorption was proposed to be due to osteoclast apoptosis. We tested this hypothesis for both the N-containing bisphosphonates alendronate and risedronate, which inhibit farnesyldiphosphate synthase and thus protein isoprenylation, and for clodronate and etidronate, which are metabolized to adenosine triphosphate (ATP) analogs. We found, in dose-response studies, that alendronate and risedronate inhibit bone resorption (in pit assays) at doses tenfold lower than those reducing osteoclast number. At an N-bisphosphonate dose that inhibited resorption and induced apoptosis, the antiapoptotic caspase inhibitor, Z-VAD-FMK, maintained osteoclast (Oc) number but did not prevent inhibition of resorption. Furthermore, when cells were treated with either alendronate alone or in combination with Z-VAD-FMK for 24 or 48 h, subsequent addition of geranylgeraniol, which restores geranylgeranylation, returned bone resorption to control levels. On the other hand, Z-VAD-FMK did block etidronate and clodronate inhibition of resorption. Moreover, in cells treated with etidronate, but not alendronate or risedronate, Z-VAD-FMK also prevented actin disruption, an early sign of osteoclast inhibition by bisphosphonates. These observations indicate that, whereas induction of apoptosis plays a major role in etidronate and clodronate inhibition of resorption, alendronate and risedronate suppression of bone resorption is independent of their effects on apoptosis. PMID- 11728927 TI - Prevention of fracture healing in rats by an inhibitor of angiogenesis. AB - Angiogenesis is considered essential to fracture healing, but its role in the healing process remains poorly understood. Angiogenesis inhibitors, which block new blood vessel formation by specifically targeting vascular cells, are currently under development for use in cancer chemotherapy, and are potentially powerful tools for defining the consequences of angiogenic impairment on fracture healing. In this study, we directly tested the effects of the angiogenesis inhibitor TNP-470 on the healing of closed femoral fractures in an established rat model system. Beginning 1 day after fracture, animals received either angiogenesis inhibitor at a therapeutically effective antitumor dose, or a weight adjusted amount of carrier vehicle. The progress of fracture healing was assessed at weekly intervals for 21 days by radiography and histology; functional assessment was carried out at day 24 by biomechanical testing. By all three criteria, treatment with the angiogenesis inhibitor completely prevented fracture healing. Formation of both callus and periosteal woven bone were suppressed, indicating that both the intramembranous and endochondral pathways of osteogenesis were affected. The resulting tissue resembled "atrophic nonunions" often seen clinically in cases of failed fracture healing, but rarely achieved in animal models. These results show that angiogenesis is essential to very early stages of fracture healing, and suggest this model system may be useful for understanding the mechanisms underlying fracture nonunions due to vascular impairment. Finally, the data raise the possibility that impairment of fracture healing may be an adverse effect of clinical treatments with antiangiogenic drugs. PMID- 11728928 TI - The positive effects of zinc on skeletal strength in growing rats. AB - The aim of the present study was to assess the skeletal effects of alimentary zinc depletion and supplementation in an animal model of intact, growing rats. The study was planned as a dose-response study. Thirty-six male Wistar rats, 4 weeks old, were divided into three groups of 12 rats each. The rats had free access to a semisynthetic diet with different amounts of zinc added. Group 1 was given a zinc-free diet containing 2 mg zinc/kg, group 2 was given a normal-zinc diet containing 47 mg zinc/kg; and group 3 was given a zinc-supplemented diet containing 60 mg zinc/kg. All animals were killed 4 weeks after initiation of the experiment and the right femora were removed. The biomechanical effects were measured at the following skeletal sites: femoral diaphysis; femoral neck; and distal femoral metaphysis. In addition, static histomorphometry was performed at the middiaphyseal region. Biomechanical testing revealed a significant zinc induced increase in bone strength at all sites investigated. It also showed that zinc influenced bone strength in a dose-dependent manner except at the distal metaphysis, where there was no significant difference between the group fed normal-zinc diet and the group fed a hyper-zinc diet. Zinc also improved the rates of growth in the rats. The body weights and length of femora increased dose dependently. Static histomorphometry showed that zinc exerted its main effect on the periosteal envelope, thereby increasing bone area, tissue area, and axial moment of inertia. We conclude that alimentary zinc supplementation in growing rats induces an increase of bone strength in both the femoral neck and the femoral diaphysis. These results further support the view that zinc has a positive effect on bone metabolism which mimics that of growth hormone (GH) or insulin-like growth factor 1 (IGF-1). PMID- 11728930 TI - Pathology of skull base tumors. AB - The multidisciplinary requirements of skull base surgery have evolved over the last 25 years. The heterogeneity of tissue types in the cranium base gives rise to a diverse group of benign and malignant neoplasms with vastly different prognoses. This article reviews the distinct clinicopathologic features of some of the unique and problematic neoplasms of this region. PMID- 11728931 TI - Radiology of skull base neoplasms. AB - Computed tomography (CT) and magnetic resonance (MR) imaging are used routinely to stage skull base neoplasms preoperatively, define the extent of the tumor, identify perineural spread, plan surgery and radiation therapy, and evaluate the postoperative patient. Arteriography and interventional radiology also play a role in the identification and treatment of certain skull base neoplasms, particularly juvenile nasopharyngeal angiofibromas and paragangliomas. The skull base can be divided into three parts: anterior, central, and posterior. Pathology differs in each region, and CT and MR imaging often demonstrate characteristic imaging features suggestive of diagnosis. PMID- 11728932 TI - Radiotherapy and radiosurgery for skull base tumors. AB - Tumors arising in the vicinity of the skull base are relatively uncommon; however, lesions that may be successfully treated by radiotherapy and radiosurgery include temporal bone chemodectomas, schwannomas, juvenile angiofibromas, pituitary adenomas, and meningiomas. This article reviews treatment techniques and results and discusses the pertinent literature. PMID- 11728933 TI - Chemotherapy for skull base cancers. AB - This article focuses on treatment options for select skull base problems that have decreased post-treatment morbidity and, in many cases, improved survival. The select skull base cancers covered include nasopharyngeal carcinoma, squamous cell carcinoma of the paranasal sinuses, sinonasal undifferentiated carcinoma, neuroendocrine carcinoma, esthesioneuroblastoma, and salivary gland carcinoma. PMID- 11728934 TI - Endonasal approaches for sinonasal and nasopharyngeal tumors. AB - This article reviews the published literature on endonasal approaches for sinonasal and nasopharyngeal tumors and synthesizes this information with the author's personal experience into a rational approach to patients with the following disorders: inverted papilloma, adenocarcinoma, hemangioendothelioma, olfactory neuroblastoma, carcinosarcoma, squamous cell carcinoma, melanoma, juvenile angiofibroma, chordoma, and chondrosarcoma. PMID- 11728935 TI - The midfacial degloving procedure for nasal, sinus, and nasopharyngeal tumors. AB - The midfacial degloving approach is more technically involved than a lateral rhinotomy and requires a basic level of proficiency and understanding of closed rhinoplasty incisions and anatomy of the nose, paranasal sinuses, and skull base structures. Current applications of the midfacial degloving procedure have allowed expansion of indications for this technique through the use of complementary endoscopic and subcranial approaches, permitting the exposure and removal of extensive skull base lesions without disfiguring facial incisions. Fundamental in these approaches is the basic midfacial degloving exposure, which is discussed in this article, along with the applications for treatment of skull base lesions. PMID- 11728936 TI - Transoral approaches to the clivus and nasopharynx. AB - This article presents the surgical anatomy of the clivus, nasopharynx, and craniocervical junction as it relates to the transoral approach to the midline basicranium. Particular attention is given to the indications, limitations, and complications of this procedure. The transpalatal approach is discussed in detail. Finally, the transmandibular approach is also briefly discussed. PMID- 11728937 TI - Craniofacial approaches to tumors of the anterior skull base. AB - An approach that combines extracranial and intracranial components-termed a craniofacial approach-allows en bloc extirpation of paranasal malignancies that abut or penetrate the skull base. When combined with radiotherapy, cure rates for such tumors rose from near zero in the 1950s to 39% to 86% by 2000, with the higher rates reflective of esthesioneuroblastomas, well-differentiated adenocarcinomas, vasoformative tumors, and meningiomas. Transfacial tumor access can involve a transnasal (endoscopic or with magnification loupes) exposure, a midfacial degloving, or a lateral rhinotomy, depending on tumor location and size. Adjunctive exposures for tumors penetrating the nasopharynx, pterygomaxillary fossa, or sphenoid include the lateral facial split and the mandibular swing. The standard transcranial accesses are a sub-basal variation of frontal craniotomy, which encompasses en bloc the superior orbital rims, the nasion, and the lower frontal bones; or, a frontotemporal craniotomy with mobilization of the lateral orbital rim and the zygoma. PMID- 11728938 TI - Pituitary surgery. AB - This article focuses on surgical indications, perioperative management, and endoscopic approaches for pituitary surgery. Also discussed are sinonasal, neurologic, and endocrine complications, which may include bleeding, sinusitis, and synechiae; cerebrospinal fluid leaks and bleeding; and diabetes insipidus and panhypopituitarism, respectively. PMID- 11728939 TI - Orbital tumors. AB - Neoplasms involving the orbit are divided into primary and secondary types. Primary orbital tumors are rare, include a wide variety of histopathologic types, are usually benign, and most often are managed surgically. Secondary orbital tumors are most often malignant, arise from the paranasal sinuses, and are managed with multiple modalities in a planned fashion. This article discusses primary and secondary orbital tumors, including their evaluation and management. PMID- 11728940 TI - Lateral craniofacial approaches to the skull base and infratemporal fossa. AB - The field of skull base surgery has expanded dramatically the surgical horizons in the treatment of skull base neoplasms, which, previously, were considered inoperable because of their locations and relation to vital structures. This article aims to offer improved understanding of surgical indications and appreciation for some of the details of surgical techniques involved. PMID- 11728941 TI - Temporal bone carcinoma. AB - Squamous cell carcinoma is the most common malignancy of the external auditory canal, middle ear, and mastoid. Surgical resection is the cornerstone of treatment, with T1 lesions of the external auditory canal treated by lateral temporal bone resection and more advanced lesions treated by subtotal or total temporal bone resection. This article summarizes clinical manifestations, differential diagnosis, management, and differences in treatment philosophy. PMID- 11728942 TI - Petroclival tumors: a synthesis. AB - This article focuses on petroclival tumors, which are rare lesions of the posterior fossa-an area that is difficult to access. Because of their location, rarity, insidious growth, and relentless natural progression toward a fatal outcome, petroclival tumors pose major management problems. With improved microsurgical techniques, however, these tumors can be approached and removed with preservation of vital neural and vascular structures. PMID- 11728943 TI - Jugulotympanic paragangliomas. AB - Jugulotympanic paragangliomas are the most common tumors of the middle ear and temporal bone. Although these larger tumors can prove to be formidable, the advent of microscopic and skull base surgical techniques has greatly enhanced the ability to treat and manage these tumors. PMID- 11728944 TI - Skull base reconstruction and rehabilitation. AB - This article separates skull base reconstruction into the surgical procedures available for immediate reconstruction of a small base defect and the secondary rehabilitative procedures that may be performed at a later date, usually for functional or cosmetic needs. PMID- 11728945 TI - Renal and cardiovascular effects of selective cyclooxygenase-2 inhibitors. AB - Selective inhibition of cyclooxygenase-2 (COX-2) was proposed as a novel anti inflammatory and analgesic treatment with a reduced profile of gastrointestinal side effects compared with conventional nonsteroidal anti-inflammatory drugs (NSAIDs). Although perceived as an inducible enzyme by inflammatory and other stimuli, COX-2 is constitutively expressed in the kidney. In this review, we focus on renal and cardiovascular (CV) physiological and pathophysiological characteristics of COX-2 and renal and CV aspects of treatment with selective COX 2 inhibitors. Both clinical and experimental studies have shown that renal and CV effects of COX-2 inhibitors are similar to those of NSAIDs. These effects include sodium, potassium, and water retention and decreases in renal function, as well as mild to modest increases in blood pressure (BP) and edema. These deleterious effects are amplified in patients with volume and/or sodium depletion. The concomitant administration of COX-2 inhibitors may destabilize BP control in hypertensive patients treated with antihypertensive agents. In contrast to the normal kidney, which could constitute a target for adverse actions of COX-2 inhibitors, recent experimental studies showed increased renal COX-2 expression in several models of renal injury, such as the remnant kidney, renovascular hypertension, and diabetes, and implicated COX-2 in the progression of renal failure. This suggests that COX-2 inhibitors may confer a renoprotective effect in diverse renal disorders. These intriguing formulations must be delineated further in appropriately designed prospective clinical trials. PMID- 11728946 TI - Renal function in relation to three candidate genes. AB - We recently found that femoral intima media thickness, as well as the incidence of hypertension, is influenced by genes encoding the angiotensin-converting enzyme (ACE; insertion/deletion [I/D]) polymorphism, alpha-adducin (Gly460Trp), and aldosterone synthase (-344C/T). By interfering with blood pressure or sodium homeostasis, these genetic polymorphisms also may change renal function. We therefore investigated serum creatinine level, calculated and measured creatinine clearances, and 24-hour urinary protein excretion in subjects previously genotyped for these three polymorphisms. The 1,454 participants drawn at random from the population (64.3% of those invited) were aged 43.4 years and included 744 women (51.2%). Blood pressure, measured by study nurses at subjects' homes, averaged 123/76 mm Hg. Mean values were 90 micromol/L for serum creatinine; 84 and 88 mL/min/1.73 m(2) for calculated and measured (n = 855) creatinine clearances, respectively; and 90 mg/d of protein for proteinuria (n = 556). The prevalence of mild renal dysfunction (creatinine clearance /=135 mm Hg) and 1 patient had diastolic hypertension (>/=85 mm Hg) at the final visit. Four patients were administered 10 mg of lisinopril; 5 patients, 20 mg; and 2 patients, 40 mg; only 1 of these patients required ultrafiltration therapy. There was a persistent antihypertensive effect over 44 hours. BP reduction was achieved without an increase in intradialytic symptomatic or asymptomatic hypotensive episodes. PRA increased in response to dialysis, as well as lisinopril. In conclusion, supervised lisinopril therapy is effective in controlling hypertension in chronic HD patients. This may be related to blockade of angiotensin II generation by kidneys despite the loss of excretory function. PMID- 11728958 TI - A malnutrition-inflammation score is correlated with morbidity and mortality in maintenance hemodialysis patients. AB - Malnutrition inflammation complex syndrome (MICS) occurs commonly in maintenance hemodialysis (MHD) patients and may correlate with increased morbidity and mortality. An optimal, comprehensive, quantitative system that assesses MICS could be a useful measure of clinical status and may be a predictor of outcome in MHD patients. We therefore attempted to develop and validate such an instrument, comparing it with conventional measures of nutrition and inflammation, as well as prospective hospitalization and mortality. Using components of the conventional Subjective Global Assessment (SGA), a semiquantitative scale with three severity levels, the Dialysis Malnutrition Score (DMS), a fully quantitative scoring system consisting of 7 SGA components, with total score ranging between 7 (normal) and 35 (severely malnourished), was recently developed. To improve the DMS, we added three new elements to the 7 DMS components: body mass index, serum albumin level, and total iron-binding capacity to represent serum transferrin level. This new comprehensive Malnutrition-Inflammation Score (MIS) has 10 components, each with four levels of severity, from 0 (normal) to 3 (very severe). The sum of all 10 MIS components ranges from 0 to 30, denoting increasing degree of severity. These scores were compared with anthropometric measurements, near-infrared-measured body fat percentage, laboratory measures that included serum C-reactive protein (CRP), and 12-month prospective hospitalization and mortality rates. Eighty-three outpatients (44 men, 39 women; age, 59 +/- 15 years) on MHD therapy for at least 3 months (43 +/- 33 months) were evaluated at the beginning of this study and followed up for 1 year. The SGA, DMS, and MIS were assessed simultaneously on all patients by a trained physician. Case-mix-adjusted correlation coefficients for the MIS were significant for hospitalization days (r = 0.45; P < 0.001) and frequency of hospitalization (r = 0.46; P < 0.001). Compared with the SGA and DMS, most pertinent correlation coefficients were stronger with the MIS. The MIS, but not the SGA or DMS, correlated significantly with creatinine level, hematocrit, and CRP level. During the 12-month follow-up, 9 patients died and 6 patients left the cohort. The Cox proportional hazard-calculated relative risk for death for each 10-unit increase in the MIS was 10.43 (95% confidence interval, 2.28 to 47.64; P = 0.002). The MIS was superior to its components or different subversions for predicting mortality. The MIS appears to be a comprehensive scoring system with significant associations with prospective hospitalization and mortality, as well as measures of nutrition, inflammation, and anemia in MHD patients. The MIS may be superior to the conventional SGA and the DMS, as well as to individual laboratory values, as a predictor of dialysis outcome and an indicator of MICS. PMID- 11728959 TI - A 5-year follow-up on antibody response after diphtheria and tetanus vaccination in hemodialysis patients. AB - Chronic renal failure is associated with a T-cell-dependent immune defect. In the past, various studies have focused on the insufficient immune response to vaccination of hemodialysis patients. An impaired vaccination response rate has been reported for vaccines against hepatitis B, influenza, tetanus, diphtheria, and others. However, no data exist on the long-term success of vaccination against tetanus and diphtheria in these patients. The aim of the present study is to investigate seroresponse to tetanus and diphtheria vaccination over a 5-year period. Antibody levels were determined by enzyme immunoassay. Antidiphtheria antibody levels of 31 hemodialysis patients were determined 5 years after vaccination. After 5 years, 10 of 31 patients (32%) had a protective antibody level against diphtheria (>/=0.1 IU/mL), whereas 12 months after vaccination, 26 of 71 patients (37%) were protected. Also, mean antibody levels significantly decreased. Furthermore, antitetanus antibody levels of 21 patients simultaneously vaccinated against tetanus and diphtheria were investigated. After 5 years, 15 of 21 patients (71%) were protected compared with 46 of 71 patients (65%) in the hemodialysis collective studied after 12 months. In the interval between 1 and 5 years after vaccination, significantly more patients in the initial nonresponder group had died than in the responder group; therefore, the overall protection rate observed in vaccinated patients increased. Our results provide evidence that during a 5-year period, antibody persistence against tetanus toxoid is better than that against diphtheria toxoid. Therefore, detection of individual antibody concentrations may be indicated to decide on revaccination. PMID- 11728960 TI - Soluble Fas is a marker of coronary artery disease in patients with end-stage renal disease. AB - Coronary artery disease (CAD) is the leading cause of death in patients with end stage renal disease (ESRD). Recent evidence suggests that the expression of Fas, a molecule implicated in the initiation of apoptosis in various cell types, is increased at sites of atherosclerotic plaques. However, the significance of plasma levels of the soluble form of Fas (sFas) and its ligand (sFas-L) as markers of atherosclerosis has yet to be defined. The present report is a cross sectional analysis of baseline data from an ongoing prospective study designed to evaluate the role of sFas and sFas-L as markers of CAD in ESRD. We evaluated the association between plasma levels of sFas and sFas-L and evidence of CAD in a cohort of 107 chronic hemodialysis patients. Plasma levels of sFas were significantly greater (P = 0.04) among subjects with (n = 64) than without evidence of CAD (n = 43). Plasma levels of sFas-L were similar in both groups. Using multivariate analysis, sFas level was found to be independently associated with CAD (P = 0.01) after adjustment for classic risk factors for CAD (hyperlipidemia, diabetes, hypertension, and smoking), markers of inflammation (C reactive protein [CRP], intercellular adhesion molecule 1), and other confounders. An increase of one quintile in plasma concentration of sFas was associated with an odds ratio for CAD of 1.64 (95% confidence interval, 1.11 to 2.41). Models that incorporated sFas were significantly better at identifying patients with CAD than models limited to classic risk factors for atherosclerosis, alone (P = 0.008) or in combination with CRP levels (P = 0.006). In summary, increased plasma levels of sFas are associated with CAD in stable patients with ESRD. These results suggest that sFas may represent a novel and independent marker of CAD. PMID- 11728961 TI - Artificial intelligence: a new approach for prescription and monitoring of hemodialysis therapy. AB - The effect of dialysis on patients is conventionally predicted using a formal mathematical model. This approach requires many assumptions of the processes involved, and validation of these may be difficult. The validity of dialysis urea modeling using a formal mathematical model has been challenged. Artificial intelligence using neural networks (NNs) has been used to solve complex problems without needing a mathematical model or an understanding of the mechanisms involved. In this study, we applied an NN model to study and predict concentrations of urea during a hemodialysis session. We measured blood concentrations of urea, patient weight, and total urea removal by direct dialysate quantification (DDQ) at 30-minute intervals during the session (in 15 chronic hemodialysis patients). The NN model was trained to recognize the evolution of measured urea concentrations and was subsequently able to predict hemodialysis session time needed to reach a target solute removal index (SRI) in patients not previously studied by the NN model (in another 15 chronic hemodialysis patients). Comparing results of the NN model with the DDQ model, the prediction error was 10.9%, with a not significant difference between predicted total urea nitrogen (UN) removal and measured UN removal by DDQ. NN model predictions of time showed a not significant difference with actual intervals needed to reach the same SRI level at the same patient conditions, except for the prediction of SRI at the first 30-minute interval, which showed a significant difference (P = 0.001). This indicates the sensitivity of the NN model to what is called patient clearance time; the prediction error was 8.3%. From our results, we conclude that artificial intelligence applications in urea kinetics can give an idea of intradialysis profiling according to individual clinical needs. In theory, this approach can be extended easily to other solutes, making the NN model a step forward to achieving artificial-intelligent dialysis control. PMID- 11728962 TI - Retrospective analysis of the effects of cisapride on the QT interval and QT dispersion in chronic hemodialysis patients. AB - Cisapride is contraindicated in patients with end-stage renal disease (ESRD) and gastrointestinal motility disorders. Ventricular arrhythmias have been associated with both cisapride and hemodialysis (HD). However, reports conflict regarding the safety of cisapride in HD patients. We undertook this study to characterize the effects of cisapride on QT intervals and QT dispersion (QTD) in HD patients. Baseline and steady-state electrocardiograms (ECGs) were retrospectively selected for calendar year 1999 for each patient administered cisapride if ECGs showed sinus rhythm, potassium level was 3.5 mEq/dL or greater, and there was no pharmacokinetic drug interaction. QT intervals were measured by two investigators, and QTDs were calculated (maximum [QT(max)] - minimum QT interval [QT(min)]). Averages between investigator measures (+/- SD), presented for each value, were evaluated using Wilcoxon's signed-rank test. Thirty-one HD patients were administered cisapride. Seventeen patients failed to meet entry criteria, and no patient had a pharmacokinetic drug interaction. In included patients (6 men, 8 women), heart rates were 86.71 +/- 20.87 beats/min at baseline and 86.57 +/- 14.23 beats/min during treatment (P = not significant). Serum potassium levels were 4.97 +/- 1.2 mEq/dL at baseline and 4.94 +/- 0.76 mEq/dL during treatment (P = not significant). Average baseline QT(max) and QT(min) were 391.07 +/- 42.43 and 330.71 +/- 40.94 milliseconds, respectively. Treatment QT(max) and QT(min) were 391.43 +/- 38.2 and 343.93 +/- 35.69 milliseconds, respectively (P = not significant for both). QTD was 60.36 +/- 17.59 milliseconds at baseline and 47.5 +/- 19.59 milliseconds during treatment (P = 0.074). Mean corrected QT (QTc) intervals increased from 426.57 +/- 26.62 to 431.71 +/- 29.98 milliseconds (P = 0.55) from baseline to treatment. No ventricular arrhythmia was observed during at least 160 days (range, 2 to 830 days) of cisapride exposure. Two patients died during this study, both of other causes 4 days after discontinuing cisapride therapy. Cisapride did not significantly increase mean QTc interval, QT(max), or QTD in patients with ESRD managed by HD when potassium levels were stable and pharmacokinetic drug interactions were avoided. PMID- 11728963 TI - Comparison of L-type and mixed L- and T-type calcium channel blockers on kidney injury caused by deoxycorticosterone-salt hypertension in rats. AB - The efficiency of calcium channel blockers (CCBs) in the treatment of chronic renal disease (CRD) is controversial. In this study, we investigated whether combined T- and L-type CCBs, using mibefradil (30 mg/kg/d), provided superior protection versus traditional L-type voltage-gated CCBs, using amlodipine (10 mg/kg/d), in the deoxycorticosterone acetate (DOCA)-salt model of high glomerular blood pressure (P(GC)) and rapidly developing kidney damage. After 4 to 5 weeks of DOCA-salt, amlodipine did not reduce proteinuria (protein, 341 +/- 90 versus 482 +/- 54 mg/24 h; P = not significant) or degree of glomerular damage (20% +/- 4% versus 28% +/- 6% damaged glomeruli; P = not significant) compared with untreated rats. Conversely, mibefradil reduced proteinuria and glomerular damage versus untreated DOCA-salt rats (protein, 244 +/- 75 mg/24 h; P < 0.02; damaged glomeruli, 11% +/- 3%; P < 0.05). Both CCBs had similar antihypertensive actions, returning blood pressure to the untreated sham value. Of note, P(GC) also was reduced by a similar extent (and to the sham value) with both mibefradil (58 +/- 2 mm Hg; P < 0.001) and amlodipine (61 +/- 2 mm Hg; P < 0.005) versus untreated DOCA-salt rats (70 +/- 1 mm Hg). This study shows that combined T- and L-type CCBs provide superior protection against CRD in the DOCA-salt model compared with L-type CCBs alone. However, this protection was not hemodynamic because similar systemic and glomerular antihypertensive responses occurred with both mibefradil and amlodipine. Although mibefradil was withdrawn from the market because of adverse drug interactions not associated with CCBs, other mixed channel blockers may provide an alternative or adjunctive therapy to angiotensin-converting enzyme inhibition in CRD. PMID- 11728964 TI - Alterations of vitronectin and its receptor alpha(v) integrin in the rat renal glomerular wall during diabetes. AB - Vitronectin, a multifunctional glycoprotein present in blood and extracellular matrix, is not only a member of the cell adhesion molecules, but also a regulator of proteolytic enzyme cascades, thereby providing a unique regulatory factor for proteolytic degradation of extracellular matrix and tissue remodeling. Vitronectin interacts with the cell surface through integrins of the alpha(v) related system. Because vitronectin and its receptor may have a role in various renal physiological and pathological processes, we evaluated their expression in renal tissues of streptozotocin-induced short- and long-term hyperglycemic rats by applying quantitative immunoelectron microscopy and Western blot analysis. Vitronectin was shown over the glomerular basement membrane (GBM) and mesangial matrix (MM), whereas alpha(v) was located along the plasma membrane of endothelial, epithelial, and mesangial cells. Although distribution patterns of vitronectin and alpha(v) integrin labeling in renal tissues from short- and long term hyperglycemic rats, as well age-matched normoglycemic rats, were similar, increases in their immunoreactive sites were detected in hyperglycemic conditions. Changes also were present in old compared with young normoglycemic animals. The diabetes-related increase in vitronectin was more significant in the GBM than MM, whereas the increase in alpha(v) integrin was as significant in podocytes as mesangial cells. Western blot analysis, performed on isolated glomerular material from normoglycemic and hyperglycemic animals, confirmed those changes. Our results suggest that vitronectin and its receptor, alpha(v) integrin, must have defined roles in molecular mechanisms involved in the pathogenesis of both diabetic and aging nephropathy. PMID- 11728965 TI - Novel mutation in the GRHPR gene in a Chinese patient with primary hyperoxaluria type 2 requiring renal transplantation from a living related donor. AB - We identified a patient with primary hyperoxaluria type 2 (PH2) showing recurrent stone formation, nephrocalcinosis, end-stage renal failure, and rapid oxalate deposition after renal transplantation from a living related donor. Urinary organic acid analysis performed after renal transplantation confirmed the diagnosis of PH2. We analyzed the glyoxylate reductase/hydroxypyruvate reductase (GRHPR) gene of the patient. DNA sequencing of all nine exons and exon-intron boundaries showed a novel homozygous mutation deleting the last two nucleotides of exon 8, ie, 862delTG. This deletion results in a frameshift and introduction of a premature stop codon at codon 310, ie, Ala310Stop. One of the patient's sisters is heterozygous for this mutation, and the other sister, who is the donor, does not have this mutation. The rapid deposition of oxalate in the transplanted kidney indicates that the kidney is not a major site of oxalate production. The more favorable long-term prognosis of PH2 needs to be reevaluated now that the molecular basis of PH2 has been established. DNA-based diagnosis will facilitate carrier detection, prenatal diagnosis, genetic counseling, and selection of living related donors. PMID- 11728966 TI - Hemodialysis central venous catheter tip fracture with embolization into the pulmonary artery. AB - We describe the first reported case of spontaneous hemodialysis catheter fracture and embolization. We also include a summary of similar cases not previously reported in the literature that appear in the Food and Drug Administration records as medical equipment malfunctions. Our patient presented with a cough and was initially treated for presumptive pneumonia. A chest radiograph indicated that the tip of the catheter had fractured and migrated into a distal branch of the pulmonary artery. The catheter was replaced, but the tip could not be retrieved because of its peripheral location. Possible causes and consequences of this underreported complication are discussed. PMID- 11728968 TI - Can the future of hemodialysis patients be predicted? PMID- 11728969 TI - T-cell-specific therapy in autoimmune glomerulonephritis. PMID- 11728970 TI - A young man with 20 years of nephrotic range proteinuria without loss of renal function. PMID- 11728971 TI - Improving prognosis for kidney disorders in the 21st century: hypertension, anemia, nutrition, and lipids: introduction. PMID- 11728972 TI - Mechanisms activated by kidney disease and the loss of muscle mass. AB - The daily turnover of cellular proteins is large, with amounts equivalent to the protein contained in 1.0 to 1.5 kg of muscle. Consequently, even a small, persistent increase in the rate of protein degradation or decrease in protein synthesis will result in substantial loss of muscle mass. Activation of protein degradation in the ubiquitin-proteasome system is the mechanism contributing to loss of muscle mass in kidney disease. Because other catabolic conditions also stimulate this system to cause loss of muscle mass, the identification of activating signals is of interest. A complication of kidney disease, metabolic acidosis, activates this system in muscle by a process that requires glucocorticoids. The influence of inflammatory cytokines on this system in muscle is more complicated, as evidence indicates that cytokines suppress the system, but glucocorticoids block the effect of cytokines to slow protein breakdown in the system. New information identifying mechanisms that activate protein breakdown and the rebuilding of muscle fibers would lead to therapies that successfully prevent the loss of muscle mass in kidney disease and other catabolic illnesses. PMID- 11728973 TI - Relative contributions of nutrition and inflammation to clinical outcome in dialysis patients. AB - Protein-energy malnutrition (PEM) is a common phenomenon in maintenance dialysis (MD) patients and a risk factor for poor quality of life and increased morbidity and mortality, including cardiovascular death, in these individuals. The association between undernutrition and adverse outcome in MD patients, which stands in contrast to that seen in the general population, has been referred to as reverse epidemiology. Measures of food intake, body composition tools, nutritional scoring systems, and laboratory values are used to assess the degree of severity of PEM, but no uniform approach is available for rating the overall severity of PEM. Epidemiologic studies suggest that inflammation is a missing link between PEM and poor clinical outcome in MD patients, and the existence of a malnutrition inflammation complex syndrome is suggested in these patients. Inflammation may be due to subclinical and clinically apparent illnesses. Some investigators suggest that PEM may predispose to illness and inflammation. There is a paucity of information concerning the effect of nutritional therapy on morbidity and mortality in MD patients. Interventional studies of the effect of nutritional support on outcome often are difficult to interpret because of small sample sizes, short duration of study, and other limitations. Large-scale, randomized, clinical trials of the effects of nutritional intake, nutritional status, and inflammation on clinical outcome are needed to define better the relationships between these factors in MD patients. PMID- 11728974 TI - Importance of low serum intact parathyroid hormone as a predictor of mortality in hemodialysis and peritoneal dialysis patients: 14 years of prospective observation. AB - Excess parathyroid hormone (PTH) has long been considered detrimental to the health of patients with end-stage renal disease. PTH has been implicated as a multisystem uremic toxin, and hyperparathyroidism can be a debilitating complication in dialyzed patients. We have studied prospectively the relationship of enrollment serum intact PTH and various demographic characteristics and other biochemical parameters to all-cause mortality in 345 hemodialysis (HD) and 277 peritoneal dialysis (PD) patients. We monitored the patients for 14 years. Observed survival and survival after adjustment for age, race, gender, months on dialysis at enrollment, diabetic status, and nutritional markers were significantly better for patients with enrollment PTH greater than 200 pg/mL than for patients with PTH 65 to 199 pg/mL and patients with PTH less than 65 pg/mL. Enrollment serum PTH was an independent predictor of survival in HD and PD patients. For HD patients, age and months on HD at enrollment were associated inversely with PTH level, whereas black race, creatinine, and phosphorus were associated directly with PTH. For PD patients, age, diabetes, and months on PD at enrollment were inverse predictors, whereas black race, albumin, creatinine, and phosphorus were associated positively with PTH. Lower than expected levels of PTH in uremic patients is associated with increased mortality. We hypothesize that inadequate protein intake or phosphorus intake or both result in impaired development of the expected secondary hyperparathyroidism and in the excess mortality risk inherent with malnutrition. PMID- 11728975 TI - Serum prealbumin predicts survival in hemodialysis and peritoneal dialysis: 10 years of prospective observation. AB - Malnutrition is a major factor contributing to the high mortality rate in hemodialysis (HD) and peritoneal dialysis (PD) patients. We and others have reported previously that single enrollment levels of serum biochemical markers, such as albumin, cholesterol, creatinine, and prealbumin, are correlated directly with mortality in HD and PD patients. We have studied prospectively the relationship of enrollment prealbumin levels, demographic characteristics, and other biochemical markers to all causes of mortality in 130 HD and 128 PD patients who were monitored for 10 years. The Kaplan-Meier method was used to compute observed survival, and the Cox proportional hazards model was used to identify independent predictors of mortality risk. For HD patients, enrollment serum prealbumin remained a strong independent predictor of long-term survival after adjusting for age, race, gender, months on dialysis, diabetic status, and other nutritional markers. In PD and HD patients, observed and adjusted survivals (after adjusting for aforementioned confounding variables) of patients with prealbumin greater than 30 mg/dL were significantly higher than survivals of patients with prealbumin less than 30 mg/dL. For HD and PD patients, age and diabetes were associated inversely with prealbumin concentration, whereas levels of albumin, creatinine, and total cholesterol were associated directly with prealbumin concentration. In this study, prealbumin was the best biochemical predictor of mortality for HD patients and a useful tool to assess nutritional risk in HD and PD patients. PMID- 11728976 TI - Achieving maximal renal protection in nondiabetic chronic renal disease. AB - A rapid global increase in the number of patients requiring renal replacement therapy necessitates that effective strategies for renal protection are developed and applied widely. We review the experimental and clinical evidence in support of individual renoprotective interventions, including angiotensin-converting enzyme therapy, control of systemic hypertension, dietary protein restriction, reduction of proteinuria, treatment of hyperlipidemia, and smoking cessation. We also consider potential future renoprotective therapies. To achieve maximal renal protection, a comprehensive strategy employing all of these elements is required. This strategy should be directed at normalizing clinical markers of renal disease to induce a state of remission. PMID- 11728977 TI - Metabolic pathogenesis of cardiorenal disease. AB - Clinical data have established microalbuminuria/proteinuria as an independent risk factor for the development and progression of renal disease in patients with either diabetes or essential hypertension. Decreased kidney function is associated with increased cardiovascular risk, even at modest reductions in estimated creatinine clearance (to approximately 60 mL/min/1.73 m(2)) or modest elevations in serum creatinine (>1.4 mg/dL). Treatment with angiotensin converting enzyme inhibitors has been shown in clinical trials to delay or stabilize the rate of progression of renal disease. Reduction in cardiovascular events, such as stroke and myocardial infarction, also has been shown in these high-risk individuals. These effects are dependent and independent of blood pressure control, suggesting a nonhemodynamic effect in blockade of the renin angiotensin system. In conjunction with other therapeutic interventions, such as dietary modification and control of serum lipids, it appears that for at least a subgroup of patients it is possible to delay or prevent progression of kidney failure. There frequently is a clustering of risk factors in these individuals, including insulin resistance, salt sensitivity, hypertension, and dyslipidemia. The mechanism of the relationship between albuminuria and cardiovascular disease is unclear but may be related to endothelial cell dysfunction. Nonetheless, the presence of microalbuminuria/proteinuria as a marker for cardiovascular disease has important implications for the identification and treatment of individuals at risk. PMID- 11728978 TI - Progression of diabetic nephropathy: role of plasma homocysteine and plasminogen activator inhibitor-1. AB - Among patients with diabetic nephropathy, the decline in glomerular filtration rate (GFR) varies substantially, ranging from 2 to 20 mL/min per year. Identification of predictors of progression in diabetic nephropathy is important. Plasma total homocysteine (tHcy) rises with urinary albumin excretion rate in diabetes, and plasminogen activator inhibitor-1 (PAI-1) has been correlated with increased matrix accumulation in various glomerulopathies. In this prospective observational cohort study, we evaluated the importance of baseline tHcy and PAI 1 as predictors of the rate of decline in GFR. Baseline tHcy and PAI-1 were measured in 157 type 1 diabetic patients with diabetic nephropathy (92 men; mean age, 41 +/- 10 years; mean diabetes duration, 27 +/- 8 years; median GFR, 80 mL/min/1.73 m(2) [range, 23 to 143 mL/min/1.73 m(2)]). Hereafter, GFR was measured yearly with a plasma clearance technique for at least 3 years (median, 7 years [range, 3.0 to 8.3 years]). The mean rate of decline in GFR was 3.7 +/- 0.3 mL/min per year. A linear regression analysis revealed a borderline significant relationship between rate of decline in GFR and tHcy (P = 0.069) and PAI-1 (P = 0.087). Analysis of the rate of decline in GFR and tertiles of tHcy and PAI-1 revealed that increasing levels of tHcy were correlated with a significantly faster decline in GFR (P = 0.025), whereas increasing levels of PAI-1 were not. After adjustment for other well-established risk factors for progression of nephropathy in a multiple linear regression analysis, however, neither tHcy levels nor PAI-1 levels were independent predictors of rate of decline in GFR. PMID- 11728979 TI - Optimal antiproteinuric dose of losartan in nondiabetic patients with nephrotic range proteinuria. AB - Although the antiproteinuric response to antihypertensive treatment is the main predictor of renoprotective efficacy in long-term renal disease, to date, dose finding studies of antihypertensives have been based only on blood pressure. We aimed to find the optimal antiproteinuric dose of the angiotensin II antagonist losartan. An open-label, dose-response study using subsequent 6-week treatment periods was performed in 10 nondiabetic patients with proteinuria of 5.8 +/- 0.8 g/d and a mean arterial pressure of 103 +/- 3.7 mm Hg without antihypertensive medication. All patients had normal to moderately impaired renal function. After the baseline period, five periods followed with a daily losartan dose of 50 mg, 100 mg, 150 mg, and 50 mg and a recovery without losartan. At the end of each period, proteinuria and mean arterial pressure were measured. The consecutive doses of losartan had a similar antihypertensive response (-11.3 +/- 2.8% by the 100-mg dose). The optimal antiproteinuric response was reached at 100 mg of losartan (-30 +/- 8%). The 50-mg dose (-13 +/- 7%) was less effective, and the 150-mg dose (-28 +/- 8%) was not more effective. A 100-mg dose of losartan is optimal for reduction of proteinuria in nondiabetic patients with nephrotic range proteinuria. PMID- 11728980 TI - Endothelial chylomicron binding is altered by interaction with high-density lipoprotein in Heymann's nephritis. AB - Very-low-density lipoprotein (VLDL) catabolism is impaired in the nephrotic syndrome, partly as a result of structural changes that impair endothelial binding in the presence of lipoprotein lipase. Previous results suggested that postsynthetic modification of VLDL by high-density lipoprotein (HDL) in nephrotic syndrome rats causes their failure to bind endothelia normally. It is unknown (1) whether the structure of secreted lipoproteins is normal before exposure to nephrotic syndrome serum and (2) whether the same structural or functional defects are imparted to chylomicrons (CMs) through their interaction with HDL from nephrotic syndrome rats. CMs were isolated from thoracic duct lymph from rats with passive Heymann's nephritis (HN) and normal controls. CMs from control rats were incubated with HDL from either HN or control rats and reisolated, and apolipoprotein E (apo E) content and endothelial binding were determined. We found that CMs secreted by HN and control rats had similar apo E/B-48 ratios. HDL from HN rats had significantly lower apo E/A-I ratios than controls. Incubation of nascent control CMs with control HDL resulted in a 4-fold increase in CM apo E content, but binding was unaffected. Incubation with HDL from HN resulted in only a 50% increase in CM apo E content but reduced binding of these treated CMs by 50% compared either with nascent control CMs or with CMs incubated with control HDL. HDL from rats with HN alters CM binding to lipoprotein lipase by a mechanism that does not involve reducing the content of apo E already present on CMs at the time of secretion. PMID- 11728981 TI - Novel erythropoiesis stimulating protein for managing the anemia of chronic kidney disease. AB - Since its introduction, recombinant human erythropoietin (rHuEPO) has become the standard of care for renal anemia. Because of its relatively short half-life, however, it generally is administered two to three times per week. Darbepoetin alfa (novel erythropoiesis stimulating protein [NESP]) is a longer acting erythropoietic agent that allows less frequent dosing to treat anemia. Decreased dosing frequency should result in enhanced patient compliance and convenience and minimize the burden of frequent administration on staff. NESP is biochemically distinct from rHuEPO, having five N-linked carbohydrate chains (two more than rHuEPO). In animal studies, NESP had a half-life approximately three times longer than rHuEPO and raised hemoglobin effectively when administered less frequently than rHuEPO. NESP has been evaluated in clinical trials of dialysis and chronic kidney disease patients. Pharmacokinetic data confirmed that patients with anemia required less frequent dosing with NESP than rHuEPO. After intravenous administration, the mean elimination half-life of NESP was 25.3 hours versus 8.5 hours for rHuEPO. In patients who are rHuEPO-naive and in patients previously managed with rHuEPO, NESP is as effective as rHuEPO for maintaining hemoglobin concentration when administered intravenously or subcutaneously at a reduced frequency of once weekly or once every other week. NESP is well tolerated and has a safety profile comparable to that of rHuEPO. There have been no reports of antibody formation associated with NESP. NESP is an important new tool for physicians to use in the treatment of anemia of chronic kidney disease. PMID- 11728982 TI - Cardiovascular disease in patients with chronic kidney disease: getting to the heart of the matter. AB - The high prevalence of cardiovascular disease (CVD) in patients with kidney disease is well described. This Canadian, multicenter, observational cohort study reports the prevalence and risk factors of CVD associated with kidney disease, in a cohort of patients with established chronic kidney disease (CKD), who are followed-up by nephrologists. This analysis sought to answer 2 questions: (1) in patients with established CKD, are the prevalence and progression of CVD accounted for by conventional or uremia-related risk factors, and (2) to what extent can progression to renal replacement therapy (RRT) be explained by CVD versus traditional risk factors for kidney disease? This study population consists of 313 patients (predominantly men) who had a mean age of 56 years and a mean creatinine clearance of 36 mL/min. Thirty percent were diabetic. The overall prevalence of CVD was 46%, and was independent of severity of kidney dysfunction (P = 0.700). The median follow-up time was 23 months, for a total of 462 patient years. We note the overall incidence of CVD events (new CVD or worsening of CVD) was 47/244 (20%). The best predictors of new CVD events among those without preexisting CVD were diabetes (odds ratio [OR] = 5.35, P = 0.018) and age (OR = 1.26, P = 0.08). In those with preexisting CVD, low diastolic pressure (DP) (OR =.72, P = 0.004) and high triglycerides (OR = 1.48, P = 0.019) at baseline were independent predictors of progression of CVD. We could not determine an independent impact of kidney function on CVD in the overall cohort. Furthermore, we determined that the presence of CVD itself confers an increased risk for progression to RRT (relative risk [RR] = 1.58, P = 0.047), adjusted for kidney function. This is the first in-depth analysis of CVD in a cohort of patients with established chronic kidney disease who are not on dialysis. The question regarding the impact of the altered biology of uremia in contributing to CVD progression remains unanswered, and clearly needs further study. However, the findings do raise the issue of whether aggressive treatment of CVD and risk factors might, in fact, reduce progression to RRT. Further large-scale, observational studies as well as interventional studies are needed to more clearly understand the complex biology of cardiovascular and kidney disease progression. PMID- 11728983 TI - Oxidative stress and inflammation in hemodialysis patients. AB - Dialysis is associated with an increased generation of oxidants, which play an important part in the development of endothelial dysfunction and atherogenesis. Markers of oxidative stress include F2-isoprostanes and ethane. Measurements in dialysis patients before dialysis showed higher levels of esterified plasma F2 isoprostanes (1.62 +/- 0.73 ng/mL) than in control subjects (0.27 +/- 0.10 ng/mL) (P < 0.001). Furthermore, levels also correlated with high plasma C-reactive protein (CRP) levels (r =.48, P = 0.015). Breath ethane levels for dialysis patients (N = 19) were 6.32 +/- 3.16 pmol/kg-min, in contrast to 3.08 +/- 1.50 pmol/kg-min in control subjects (N = 11, P < 0.005). Analysis to investigate the relationship between CRP levels and outcome indicated that there was a significant difference in mortality rate over a 3-year period between patients with low and high CRP values (P < 0.001). Patients with high CRP (> 16.8 mg/L) levels were more than twice as likely to die as patients with low CRP levels (relative risk [RR] = 2.16; 95% confidence interval [CI], 1.50-3.09). CRP values were a significant predictor of mortality even after controlling for diabetes, albumin, ferritin, and age at commencement of dialysis. The RR for CRP after adjustment was 1.58 (95% CI, 1.06-2.34, P = 0.024). There were no significant interactions between CRP and other predictors of mortality, indicating that high CRP levels have an additive effect on the mortality risk. These findings show that hemodialysis patients are exposed to both oxidative stress and inflammation. PMID- 11728984 TI - Effects of erythropoietin and aminoguanidine on red blood cell deformability in diabetic azotemic and uremic patients. AB - Impaired red blood cell-deformability (RBC-df) is noted in patients with diabetes and may play a role in the pathogenesis of microvasculopathy and nephropathy. We report the effects of erythropoietin (EPO) alone and combined with aminoguanidine (AG) for 1 year on RBC-df in predialysis patients (P-DPs) with renal insufficiency and in end-stage renal disease (ESRD) patients on maintenance hemodialysis (DPs). Nine P-DPs who received EPO 50 U/kg by subcutaneous injection 3 times per week are compared with 5 P-DPs treated without EPO (mean serum creatinine 4.1 +/- 0.1 versus 4.2 +/- 0.6 mg/dL, respectively). Twelve DPs (Kt/V = 1.5 +/- 0.1) were studied. Six DPs received AG 200 mg/every other day by mouth and EPO 50 U/kg by intravenous (IV) injection, and 6 DPs received EPO (50 U/kg) and placebo and served as control. RBC-df improved significantly in 9 P-DPs treated with EPO at 6 months (from 2.7 +/- 0.1 to 1.6 +/- 0.2, P = 0.005). This positive effect was sustained at 12 months (P = 0.005); there was no change in RBC-df in P-DPs receiving usual care without EPO. RBC-df improved significantly and progressively at 6 and 12 months in DPs treated with EPO and AG (from 2.2 +/- 0.2 to 1.8 +/- 0.2; P = 0.01; 1.2 +/- 0.1; P = 0.001, respectively); there was limited improvement in RBC-df in DPs treated with EPO and placebo. We conclude that EPO treatment significantly improved RBC-df in diabetic P-DPs, but EPO alone has no significant effect on RBC-df after 12 months in diabetic DPs. The combination of EPO and AG restores RBC-df to near-normal levels in diabetic DPs. We speculate that the effect of EPO on RBC-df seen in P-DPs and DPs is related to increased synthesis and influx of new and younger RBCs. AG may confer protection of RBCs in DPs by blocking advanced glycosylated end-product (AGE) formation. PMID- 11728985 TI - Biochemical characterization of bona fide polycystin-1 in vitro and in vivo. AB - The most common form of autosomal dominant polycystic kidney disease (PKD) results from mutation of the PKD1 gene on chromosome 16p13.3. The gene encodes a 14-kb messenger RNA that is predicted to express a 462-kd membrane protein. The gene product, polycystin-1, has a large extracellular portion composed of a novel combination of protein-protein interacting domains and is postulated to be a plasma membrane receptor involved in cell-cell/matrix interactions. However, slow progress has been made in the characterization of polycystin-1 or the determination of its function. In fact, the protein is expressed at very low levels in tissues and cell lines and previous efforts directed at expression of recombinant protein had been largely unsuccessful. We have recently developed constructs of full-length human PKD1 complementary (cDNA) that can be expressed in both a stable and transient fashion in mammalian cells. We used these systems to characterize our antibodies and to track the protein in vivo. We report here the first biochemical characterization of recombinant polycystin-1 and show that the protein is a 520-kd glycosylated polypeptide with an unglycosylated core of 460 kd. Subcellular fractionation as well as biotinylation studies confirmed that the protein is plasma-membrane associated. Furthermore, we show that the recombinant protein localizes to cell-cell junctions in polarized madin darby canine kidney cells as revealed by indirect immunofluorescence. Our data represent the first characterization of polycystin-1 performed under highly controlled conditions. PMID- 11728986 TI - Strategies to minimize bone disease in renal failure. AB - The skeletal disorders associated with renal insufficiency result from alterations in calcium, phosphorus, and vitamin D metabolism. Each requires intervention to prevent and control the problem. Hyperparathyroidism and its treatment can also result in extraskeletal complications. To prevent the development of parathyroid hyperplasia and the skeletal complications of chronic kidney disease, it is desirable to initiate interventions early in the course of kidney disease; however, many patients present with established hyperparathyroidism and additional strategies are necessary to suppress hyperparathyroidism. Mainstays of this approach are the control of phosphorus and the use of vitamin D analogs. Phosphorus control requires the use of phosphate binders, preferably non-calcium-containing binders, to prevent intestinal phosphorus absorption. Vitamin D analogs are used to suppress hyperparathyroidism and have the potential to have lesser toxicity than calcitriol. Paricalcitol is the most widely used vitamin D analog in this country and it effectively suppresses hyperparathyroidism with only minimal effects on calcium and phosphorus. A substantial body of data in experimental animals supports the use of paricalcitol as a preferential therapeutic agent. Recently, an additional vitamin D sterol, doxercalciferol, has been introduced, which is metabolized to 1,25-dihydroxyvitamin D(2). Although initially thought to have lesser toxicity than its vitamin D(3) counterpart, recent studies have not provided support for a major difference in this regard. Doxercalciferol is also effective in lowering parathyroid hormone (PTH), though hypercalcemia in hyperphosphatemic episodes occurred relatively frequently during the clinical studies. As these therapeutic strategies are undertaken, it is important not to oversuppress PTH and decrease bone turnover to abnormally low levels because of the risk for adynamic renal bone disease. It is possible that when bone turnover is abnormally low, the extraskeletal deposition of calcium in blood vessels and other tissues is enhanced. Accordingly, constant monitoring is required during treatment, with emphasis on minimizing the calcium load, and, if monitored correctly, a satisfactory control of hyperparathyroidism may be achieved with the agents currently available. PMID- 11728988 TI - Iron sucrose or ferric gluconate? PMID- 11728992 TI - Recurrence of Wegener's granulomatosis 13 years after renal transplantation. AB - Wegener's granulomatosis (WG) can cause renal failure, requiring long-term renal replacement therapy. Renal transplantation in patients with WG is successful, but the risk for recurrence of the disease necessitates continued vigilance. We report a patient that originally presented with acute renal failure secondary to a pauci-immune focal necrotizing crescentic glomerulonephritis. Subsequent nasal involvement and serologic tests for antineutrophil cytoplasmic antibodies suggested a diagnosis of WG. PMID- 11728993 TI - Acute tubulointerstitial nephritis after wasp stings. AB - A 61-year-old Caucasian man presented with acute renal failure after multiple wasp stings. The patient required dialysis support temporarily. Work-up failed to show rhabdomyolysis or hemolysis and a kidney biopsy revealed acute allergic interstitial nephritis. The patient's renal function recovered completely after a short course of steroid therapy. Acute renal failure after wasp stings is typically caused by acute tubular necrosis in the setting of hemolysis or rhabdomyolysis. Compared with previously reported cases of acute renal failure associated with bee stings, our patient is unique in that his renal failure was caused by a hypersensitivity reaction apparently to the wasp venom. PMID- 11728994 TI - A case of familial Mediterranean fever with amyloidosis as the first manifestation. AB - We describe a 22-year-old Turkish woman with nephrotic syndrome who had a history of acute myelocytic leukemia. After careful clinical evaluation, the patient underwent a renal biopsy. Light microscopic examination showed deposition of Congo-positive material both in the mesangium and around the small vessels. By histochemical analyses, the deposited material was proved to be amyloid A (AA). Because the patient's history did not reveal any event that might explain the development of secondary amyloidosis, she was screened for mutations causing familial Mediterranean fever (FMF) and was found to be homozygous for the M694V mutation by denaturing gradient gel electrophoresis. We recommend that FMF Phenotype II and the development of amyloid nephropathy, before or without other symptoms of FMF, should be kept in mind in the face of unexplained proteinuria/amyloidosis, especially in high-risk ethnic groups. PMID- 11728995 TI - Interferon-alpha in combination with ribavirin as initial treatment for hepatitis C virus-associated cryoglobulinemic membranoproliferative glomerulonephritis. AB - Mixed cryoglobulinemia (MC) and glomerulonephritis are the most important extrahepatic manifestations of chronic hepatitis C virus (HCV) infection. In HCV infected patients with MC, renal involvement worsens the overall prognosis because of a high incidence of infection or cardiovascular disease. The relationship between MC and HCV infection has prompted the use of antiviral therapy. Two patients with chronic HCV infection, type-II MC and membranoproliferative glomerulonephritis (MPGN), presenting as nephrotic syndrome were treated with interferon (IFN)-alpha (3 MU 3 times per week) and ribavirin (15 mg/kg daily) for 6 months. Laboratory tests included measurement of anti-HCV antibodies, HCV RNA, and HCV genotyping, and characterization of circulating cryoglobulins. A pretreatment renal biopsy was performed, and the histopathologic lesions were scored according to the index of disease activity. Viremia and cryoglobulinemia were suppressed in both patients. However, a complete remission of proteinuria was observed in 1 patient only. The evaluation of the renal biopsy specimens revealed a mild MPGN (activity score: 5/24) in the patient with remission of proteinuria and a severe MPGN (activity score: 15/24) in the patient who maintained a nephrotic-range proteinuria. Although a fully satisfactory treatment is not yet available, we feel that a reasonable therapeutic strategy for HCV-infected patients with MC nephritis could be as follows: (1) antiviral treatment alone for patients with a low-grade kidney involvement, and (2) a short term course of steroids and cytotoxic drugs followed by antiviral therapy for acute exacerbations and/or rapidly progressive GN. PMID- 11728996 TI - Interstitial nephritis, hepatic failure, and systemic eosinophilia after minocycline treatment. AB - This report describes a 15-year-old white boy who presented with fever, back pain, a disseminated exanthematous rash, renal failure, and hepatopathy 3 weeks after the initiation of oral minocycline therapy for facial acne. Marked peripheral and urine eosinophilia were noted. A bone marrow aspiration showed more than 50% eosinophils without any evidence of malignancy, and a simultaneous kidney biopsy showed acute interstitial nephritis (AIN). The patient's symptoms and laboratory findings improved after high-dose steroid therapy was initiated, worsened when it was withheld, and improved again after it was reinitiated in view of the biopsy findings. The patient recovered completely, and steroids were tapered to discontinuation over 3 months. Over a year later, the patient's peripheral blood mononuclear cells (PBMCs) were cultured for 2 weeks in the presence or absence of minocycline ex vivo, and minocycline was found to induce the emergence of CD4(+) cells after 1 week in culture. In conclusion, this article shows for the first time several new aspects of minocycline-induced morbidity: renal and hepatic failure can occur together, and AIN and elevated blood eosinophil counts can be accompanied by marked bone marrow eosinophilia, suggesting a systemic allergic response as the underlying pathomechanism. Furthermore, the initial phase of such a response appears to involve CD4(+) T cells detectable ex vivo. Lastly, high-dose treatment with corticosteroids appears to be beneficial in this setting. PMID- 11728997 TI - Combined medical surgical therapy for pulmonary mucormycosis in a diabetic renal allograft recipient. AB - Mucormycosis is a rare opportunistic infection that complicates chronic debilitating diseases and immunosuppressed solid-organ transplant recipients. We present a case of life-threatening pulmonary mucormycosis in a diabetic renal allograft recipient who survived with reasonable renal function. Early recognition of this entity and prompt use of bronchoalveolar lavage (BAL) are critical to the outcome. Antifungal therapy combined with early surgical excision of infected, necrotic tissue appears to be the preferred course of action. Judicious withholding of immunosuppressants until fungemia cleared did not jeopardize allograft function. PMID- 11728998 TI - Majocchi's granuloma and posttransplant lymphoproliferative disease in a renal transplant recipient. AB - Renal transplant recipients are predisposed to infection and malignancy because of underlying long-term immunosuppressive therapy. In this case report, a renal transplant patient with coexisting Trichophyton rubrum granuloma (Majocchi's granuloma) and posttransplant lymphoproliferative disease (PTLD) is presented, showing the undesirable effects of heavy immunosuppression. Majocchi's granuloma was probably associated with PTLD as a reflection of overimmunosuppression. PMID- 11728999 TI - Polyglandular endocrine failure in a patient with amyloidosis secondary to familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is 1 of the major causes of secondary amyloidosis. Renal involvement is the main clinical complication and it mostly presents with nephrotic syndrome and chronic renal failure. Although deposition of amyloid has been reported in several endocrine glands such as the adrenal, thyroid, and testes, clinically significant functional impairment is uncommon. Herein, we describe a patient in whom the diagnosis of FMF was based on molecular screening and who presented with recurrent hypoglycemic attacks and extensive amyloid deposition affecting various organ function including adrenal, thyroid, parathyroid, testes, intestinal system, and the heart. PMID- 11729000 TI - Long-term renal transplant management. Introduction. PMID- 11729001 TI - Evolution of immunosuppression and continued importance of acute rejection in renal transplantation. AB - As steady improvement in short-term kidney graft survival and long-term outcomes prolongs the lives of transplant patients, responsibility for their care is shifting away from transplant specialists and into the hands of community nephrologists. Therefore, community nephrologists need to have a deeper understanding of immunosuppressive therapies than ever before. Pharmacologic immunosuppression has been continuously evolving over the past two decades. Azathioprine was introduced in the early 1960s. Introduction of cyclosporine (CsA) in 1983 revolutionized short-term outcomes after renal transplantation. The first monoclonal antibody immunosuppressant, OKT3, was introduced in 1986. The 1990s saw the introduction of a number of important new agents, including mycophenolate mofetil (MMF), tacrolimus, and a microemulsion CsA, as well as two new monoclonal antibodies. Combinations of these new agents, along with improving clinical care, have produced 1-year patient survival approaching 100% and graft survival exceeding 90%. The newest class of agents, the first of which is sirolimus, is called target of rapamycin (TOR) inhibitors and is used with CsA for maintenance therapy. Immunosuppressive drug therapy after kidney transplantation continues to evolve. There is a variety of pharmacologic combinations from which to choose, based on immunologic risk and side effect profiles. As new regimens are developed, ongoing communications between the transplant center and community nephrologists will be required to implement therapeutic changes and optimize patient care successfully. PMID- 11729002 TI - General health management and long-term care of the renal transplant recipient. AB - The steady improvement in short-term success rates in renal transplant patients has translated into better long-term success rates and a large number of patients with long-functioning renal transplants. The necessity for the lifelong administration of immunosuppressive medications to prevent rejection, coupled with the presence in many patients of a variety of other medical problems dating from the period of renal insufficiency prior to the time of renal transplantation, has created a large group of patients with a unique and complex set of long-term medical care needs. Due to the constraints of managed care, considerations of geography, or patient preference, the long-term care of an increasing number of renal transplant recipients has shifted away from the transplant center to the community-based nephrologist or internist. For optimal care to be delivered, it is important that the physicians managing these patients be cognizant of the complex and interacting medical issues involved in their care. Appropriate management can significantly prolong the life of the allograft as well as that of the patient. Guidelines for understanding and managing some of the more important and common general medical problems facing the long-term renal transplant recipient (eg, infectious complications, cardiovascular disease, hypertension, diabetes, hyperlipidemia, malignancy, pregnancy, bone disease, dental care, preventive care) are addressed in this section. PMID- 11729003 TI - Maintenance immunosuppression in the renal transplant recipient: an overview. AB - Managing maintenance immunosuppressive regimens after kidney transplantation is often challenging and confusing, requiring careful attention to efficacy, dosing, adverse effects, and costs of multiple medications. Most protocols combine a primary immunosuppressant (cyclosporine or tacrolimus) with one or two adjunctive agents (azathioprine, mycophenolate mofetil, sirolimus, corticosteroids). Avoiding drug-drug interactions is a major part of effective immunosuppressant management, and special situations (eg, pregnancy, intravenous dosing, caring for minority patients) can prove especially daunting. This review summarizes available data regarding current practices in maintenance immunosuppression, emphasizing issues that arise in day-to-day management of renal transplant recipients. PMID- 11729004 TI - Cardiovascular disease and the renal transplant recipient. AB - Cardiovascular complications contribute to a significant proportion of the morbidity and mortality in renal transplant patients. Underlying disease states such as diabetes and hypertension as well as risk factors associated with chronic dialysis may cause many patients to have established coronary artery and peripheral vascular disease at the time of transplantation. Progression or new onset of disease can occur after transplantation due to the continued presence of risk factors for cardiovascular disease. The benefit of modification of these risk factors such as hypertension and hyperlipidemia has been well established in the general population and has more recently been explored in the renal transplant population, although long-term studies documenting an improvement in morbidity and mortality are not available. This article focuses on the potential benefit of modification of risk factors in this setting. PMID- 11729005 TI - Long-term kidney transplant survival. AB - With improvements in short-term kidney graft survival, focus has shifted towards long-term survival. There has also been a substantial improvement in long-term survival as measured by kidney half-life. Long-term graft failure is secondary to chronic allograft nephropathy (CAN), recurrent disease, and death with a functioning graft. CAN is secondary to a combination of chronic rejection, chronic cyclosporine toxicity, and/or donor kidney disease. Risk factors for chronic rejection have been attributed to both immunological and nonimmunological causes. With a marked reduction in acute rejection rates-an important risk factor for CAN-there is a substantial improvement in kidney half-life. There are still nonimmunological factors, such as donor age, that adversely affect long-term graft survival. In addition, African-American recipients continue to have a shorter graft half-life. Recurrent disease is becoming an important cause of late graft failure. Despite the introduction of various potent immunosuppressive agents, there has been little or no impact on the prevalence as well as progression of recurrent diasease. With the reduction of acute rejection rates and improved short- and long-term graft survival, further improvements of long term graft survival will be an important focus in the 21st century. PMID- 11729006 TI - Long-term management of the renal transplant recipient: optimizing the relationship between the transplant center and the community nephrologist. AB - Rapid growth in the number of kidney transplant recipients along with improved viability of transplanted organs and increased patient survival have all led to the need for effective long-term management of these patients. The increasing numbers of transplants and the duration of posttransplant medical care can overwhelm the resources of a transplant center. These factors highlight the need for optimizing the relationship between the transplant center and the community nephrologist. There are several factors that affect the timing for transitioning patients from the transplant center to the community nephrologist and no standard timing has yet been established. Continued management of pretransplant comorbid conditions is important as are monitoring for adverse effects of failure of immunosuppressive therapy and the development of posttransplant complications. An array of testing that can be useful in monitoring these patients as well as the suggested frequency for their use are reviewed. Guidelines for the optimal interaction between the transplant center and the community nephrologist are provided to include circumstances concerning adjustment or conversion of immunosuppressive medications, evidence of allograft dysfunction, and the development of malignancy. PMID- 11729007 TI - A call to action: overcoming anxiety through active coping. PMID- 11729008 TI - The fate of integrated treatment: whatever happened to the biopsychosocial psychiatrist? AB - OBJECTIVE: The authors suggest that pharmacotherapy and psychotherapy, the major treatment modalities in psychiatry, have become fragmented from one another, creating an artificial separation of the psychosocial and biological domains in psychiatry. METHOD: After a brief discussion of the economic factors influencing this trend, the authors provide a selective overview of recent research. In the absence of systematic empirical data regarding which patients and which conditions might benefit from integrated treatment by one psychiatrist, the authors propose specific clinical situations that call for such integration and also discuss concerns about cost-effectiveness. RESULTS: Recent research suggests that combining psychotherapy and pharmacotherapy may have advantages over either treatment alone in certain clinical situations involving specific disorders. While few of the studies on combined treatment have tested whether a one-person or two-person model of treatment provision is more effective, there are a number of advantages to the one-person treatment model in which a psychiatrist conducts the psychotherapy and prescribes medication for the same patient. CONCLUSIONS: The authors suggest that further research is needed to clarify the optimal situations for the one-person model of integrated treatment and also propose systematic teaching of integrated treatment in all residency training programs. PMID- 11729009 TI - Husbands and wives. PMID- 11729010 TI - Successful multimodality treatment of severe behavioral disturbance in a patient with advanced Huntington's disease. PMID- 11729011 TI - Carl Gustav Jung, M.D., 1875-1961. PMID- 11729012 TI - Efficacy of sertraline in preventing relapse of posttraumatic stress disorder: results of a 28-week double-blind, placebo-controlled study. AB - OBJECTIVE: The study examined the efficacy of sertraline, compared with placebo, in sustaining improvement and preventing relapse over 28 weeks in patients with posttraumatic stress disorder (PTSD) who had completed a 12-week double-blind, placebo-controlled acute treatment study and a subsequent 24-week open-label study of continuation treatment with sertraline. METHOD: Ninety-six patients were randomly assigned, in a double-blind design, to 28 weeks of maintenance treatment with sertraline (50-200 mg, N=46; 78% were women) or placebo (N=50; 62% were women). Measures used in biweekly assessments included the Clinician-Administered PTSD Scale, the Impact of Event Scale, and the Clinical Global Impression severity and improvement ratings. Kaplan-Meier analyses were used to estimate time to discontinuation from the study due to relapse, relapse or study discontinuation due to clinical deterioration, and acute exacerbation. RESULTS: Continued treatment with sertraline yielded lower PTSD relapse rates than placebo (5% versus 26%). Patients who received placebo were 6.4 times as likely to experience relapse as were patients who received sertraline. Kaplan-Meier analyses confirmed the protective effect of sertraline in significantly extending time in remission. The ability of sertraline to sustain improvement was comparable across the three core PTSD symptom clusters (reexperiencing/intrusion, avoidance/numbing, and hyperarousal). A regression analysis found early response during acute treatment to be associated with a more than 16-fold reduced risk of relapse after placebo substitution. Sertraline, at a mean endpoint dose of 137 mg, was well tolerated, with no sertraline-related adverse events observed at a rate of 10% or higher. CONCLUSIONS: The results provide evidence for the ability of sertraline both to sustain improvement in PTSD symptoms and to provide prophylactic protection against relapse. PMID- 11729013 TI - Efficacy and safety of paroxetine treatment for chronic PTSD: a fixed-dose, placebo-controlled study. AB - OBJECTIVE: This study evaluated the efficacy and safety of paroxetine for the treatment of patients with chronic posttraumatic stress disorder (PTSD). METHOD: Outpatients with chronic PTSD according to DSM-IV criteria and a score of 50 or more on the Clinician-Administered PTSD Scale, part 2, were randomly assigned to take placebo (N=186), 20 mg/day of paroxetine (N=183), or 40 mg/day of paroxetine (N=182) for 12 weeks. Efficacy was assessed by examining the change in total score from baseline to endpoint on the Clinician-Administered PTSD Scale, part 2, and rates of response ("very much improved" or "much improved") for global improvement on the Clinical Global Impression scale. RESULTS: Paroxetine-treated patients in both dose groups demonstrated significantly greater improvement on primary outcome measures compared to placebo-treated patients in the intent-to treat analysis. Moreover, paroxetine treatment resulted in statistically significant improvement compared to placebo on all three PTSD symptom clusters (reexperiencing, avoidance/numbing, and hyperarousal), social and occupational impairment, and comorbid depression. Paroxetine was effective for both men and women. Treatment response did not vary by trauma type, time since trauma, or severity of baseline PTSD or depressive symptoms. Both doses were well tolerated. CONCLUSIONS: Doses of 20 and 40 mg/day of paroxetine are effective and well tolerated in the treatment of adults with chronic PTSD. PMID- 11729014 TI - An effect-size analysis of the relative efficacy and tolerability of serotonin selective reuptake inhibitors for panic disorder. AB - OBJECTIVE: Serotonin selective reuptake inhibitors (SSRIs) are now considered the first-line pharmacotherapy for panic disorder. The preferential use and the presumption of greater tolerability of SSRIs relative to older agents, such as tricyclic antidepressants, occurred without direct comparisons between the two classes of medication. In this study the authors used an effect-size analysis to provide an initial comparison. METHOD: The authors conducted an effect-size analysis of 12 placebo-controlled, efficacy trials of SSRIs for panic disorder and compared these results to findings obtained in a recent meta-analysis of non SSRI treatments for panic disorder. RESULTS: The mean effect size for acute treatment outcome for SSRIs relative to placebo was 0.55, not significantly different from that for antidepressants in general (0.55) and for imipramine in particular (0.48). More recent studies of SSRIs, and studies using larger samples, were associated with lower effect sizes. No significant differences were found in dropout rates between those taking SSRIs and those taking older agents during acute treatment. CONCLUSIONS: An effect-size analysis of controlled studies of treatments for panic disorder revealed no significant differences between SSRIs and older antidepressants in terms of efficacy or tolerability in short-term trials. An inverse relationship was evident between sample size and effect size for SSRIs. Early studies of small samples may have led to initial overestimations of the efficacy of SSRIs for panic disorder. PMID- 11729015 TI - Emotion-focused psychotherapy for patients with panic disorder. AB - OBJECTIVE: Recent studies have suggested that most patients treated for panic disorder receive forms of psychotherapy other than cognitive behavior therapy, even though there is little information about the efficacy of such treatments or how they compare to proven active treatments. The authors compared one of these other forms, emotion-focused psychotherapy (given to 30 patients with panic disorder), to results obtained with recommended standard treatment (either cognitive behavior therapy [N=36] or imipramine [N=22]). The authors also compared emotion-focused psychotherapy to results obtained in subjects given pill placebo (N=24). METHOD: Subjects met DSM-IV criteria for panic disorder with no more than mild agoraphobia. Treatment consisted of approximately 3 months of weekly visits followed by 6 monthly maintenance visits. Assessments were conducted after each treatment phase and at a follow-up visit after 6 months of no treatment. RESULTS: Emotion-focused psychotherapy was less effective for symptoms of panic disorder than treatment with either cognitive behavior therapy or imipramine; results obtained with emotion-focused psychotherapy after the acute and maintenance phases were similar to those seen with placebo. Treatment expectations were not different among the different groups. Patients receiving emotion-focused psychotherapy had the highest completion rate. CONCLUSIONS: The results suggest that emotion-focused psychotherapy (a supportive form of psychotherapy) has low efficacy for the treatment of panic disorder. However, emotion-focused psychotherapy may be superior to medical management in helping patients stay in treatment. PMID- 11729016 TI - Impact of generalized social anxiety disorder in managed care. AB - OBJECTIVE: The authors determined the costs associated with generalized social anxiety disorder in a managed care setting. METHOD: A three-phase mail and telephone survey was conducted from July to October 1998 in two outpatient clinics of a large health maintenance organization (HMO). The survey assessed direct costs, indirect costs, health-related quality of life, and clinical severity associated with generalized social anxiety disorder, both alone and with comorbid psychopathology. RESULTS: The weighted prevalence rate of current generalized social anxiety disorder was 8.2%. In the past year, only 0.5% of subjects with generalized social anxiety disorder had been accurately diagnosed. Yet 44.1% had a mental health specialty visit or had been prescribed an antidepressant, and psychiatric comorbidity was found in 43.6%. Noncomorbid generalized social anxiety disorder was associated with significantly lower health-related quality of life, work productivity, and earnings and greater utilization of health services; generalized social anxiety disorder with comorbid psychopathology was even more disabling. Suicide was attempted by 21.9% of subjects with noncomorbid generalized social anxiety disorder. Persons with average-severity generalized social anxiety disorder had probabilities of graduating from college that were 10 percentage points lower, earned wages that were 10% lower, and had probabilities of holding a technical, professional, or managerial job that were 14 percentage points lower than the comparison group. CONCLUSIONS: In a community cohort of HMO members, generalized social anxiety disorder was rarely diagnosed or treated despite being highly prevalent and associated with significant direct and indirect costs, comorbid depression, and impairment. PMID- 11729017 TI - Placebo-controlled trial of sertraline in the treatment of children with generalized anxiety disorder. AB - OBJECTIVE: The study compared the safety and efficacy of sertraline, a selective serotonin reuptake inhibitor, and placebo in the treatment of generalized anxiety disorder in children and adolescents. METHOD: The study subjects were 22 children and adolescents age 5-17 years who met the DSM-IV criteria for generalized anxiety disorder according to the Anxiety Disorders Interview Schedule for Children-Revised and who had a Hamilton Anxiety Rating Scale score > or = 16. The patients underwent a 2-3-week prestudy evaluation period, followed by a 9-week double-blind treatment phase in which they were randomly assigned in blocks of four to receive either sertraline or pill placebo. The maximum dose of sertraline was 50 mg/day. Primary outcome measures were the Hamilton anxiety scale and the Clinical Global Impression scale. RESULTS: The Hamilton anxiety scale total score, psychic factor, and somatic factor and the Clinical Global Impression severity and improvement scales showed significant differences with treatment in favor of sertraline over placebo beginning at week 4. Self-report measures reflected these results at the end of treatment. CONCLUSIONS: The results of this double-blind, placebo-controlled trial suggest that sertraline at the daily dose of 50 mg is safe and efficacious for the treatment of generalized anxiety disorder in children and adolescents. PMID- 11729018 TI - Low level of brain dopamine D2 receptors in methamphetamine abusers: association with metabolism in the orbitofrontal cortex. AB - OBJECTIVE: The role of dopamine in the addictive process (loss of control and compulsive drug intake) is poorly understood. A consistent finding in drug addicted subjects is a lower level of dopamine D2 receptors. In cocaine abusers, low levels of D2 receptors are associated with a lower level of metabolism in the orbitofrontal cortex. Because the orbitofrontal cortex is associated with compulsive behaviors, its disruption may contribute to compulsive drug intake in addicted subjects. This study explored whether a similar association occurs in methamphetamine abusers. METHOD: Fifteen methamphetamine abusers and 20 non-drug abusing comparison subjects were studied with positron emission tomography (PET) and [11C]raclopride to assess the availability of dopamine D2 receptors and with [18F]fluorodeoxyglucose to assess regional brain glucose metabolism, a marker of brain function. RESULTS: Methamphetamine abusers had a significantly lower level of D2 receptor availability than comparison subjects (a difference of 16% in the caudate and 10% in the putamen). D2 receptor availability was associated with metabolic rate in the orbitofrontal cortex in abusers and in comparison subjects. CONCLUSIONS: Lower levels of dopamine D2 receptor availability have been previously reported in cocaine abusers, alcoholics, and heroine abusers. This study extends this finding to methamphetamine abusers. The association between level of dopamine D2 receptors and metabolism in the orbitofrontal cortex in methamphetamine abusers, which replicates previous findings in cocaine abusers, suggests that D2 receptor-mediated dysregulation of the orbitofrontal cortex could underlie a common mechanism for loss of control and compulsive drug intake in drug-addicted subjects. PMID- 11729019 TI - Developmental sequence from disruptive behavior diagnosis to adolescent alcohol dependence. AB - OBJECTIVE: The authors sought to clinically describe the relationship of disruptive behavior disorders with both alcohol dependence and the use of a variety of substances. METHOD: The Child Semi-Structured Assessment for the Genetics of Alcoholism was used to collect data on 54 adolescents with a diagnosis of alcohol dependence. The frequency and age at onset of the disruptive behavior disorder diagnoses were examined as well as age at first use of alcohol, tobacco, marijuana, and other street drugs. RESULTS: Nearly three-quarters of the alcohol-dependent adolescents had at least one disruptive behavior disorder diagnosis. Attention deficit hyperactivity disorder (ADHD) typically occurred first, followed by conduct disorder. Substance use began with alcohol or tobacco, followed by marijuana and then other street drugs. Alcohol dependence began significantly later than the onset of either ADHD or conduct disorder and significantly later than the first use of tobacco. CONCLUSIONS: Disruptive behavior diagnoses, particularly conduct disorder, typically precede the initiation of use of a variety of substances that, in turn, precede the diagnosis of alcohol dependence in adolescents. PMID- 11729020 TI - Ethnic disparities in unmet need for alcoholism, drug abuse, and mental health care. AB - OBJECTIVE: Recent policy has focused on documenting and reducing ethnic disparities in availability and quality of health care. The authors examined differences by ethnic status in unmet need for alcoholism, drug abuse, and mental health treatment. METHOD: Data were from a follow-up survey of adult respondents to a 1996-1997 national survey. Non-Hispanic whites, African Americans, and Hispanics were compared in access to alcoholism and drug abuse treatment and mental health care (primary or specialty), unmet need for care, satisfaction with care, and use of active treatment for alcoholism, drug abuse, and mental health problems in the prior 12 months. RESULTS: A total of 31.9% of whites, 28.1% of African Americans, and 30.1% of Hispanics had some alcoholism, drug abuse, and mental health care, mostly in primary care. Among those with perceived need, compared to whites, African Americans were more likely to have no access to alcoholism, drug abuse, or mental health care (25.4% versus 12.5%), and Hispanics were more likely to have less care than needed or delayed care (22.7% versus 10.7%). Among those with need, whites were more likely than Hispanics or African Americans to be receiving active alcoholism, drug abuse, or mental health treatment (37.6% versus 22.4%-25.0%). CONCLUSIONS: The authors document greater unmet need for alcoholism and drug abuse treatment and mental health care among African American and Hispanics relative to whites. New policies are needed to improve access to and quality of alcoholism, drug abuse, and mental health treatment across diverse populations. PMID- 11729021 TI - Cannabis abuse as a risk factor for depressive symptoms. AB - OBJECTIVE: This study sought to estimate the degree to which cannabis abuse is a risk factor for depressive symptoms rather than an effort to self-medicate depression. METHOD: Participants (N=1,920) in the 1980 Baltimore Epidemiologic Catchment Area (ECA) study who were reassessed between 1994 and 1996 as part of a follow-up study provided the data. The analysis focused on two cohorts: those who reported no depressive symptoms at baseline (N=849) and those with no diagnosis of cannabis abuse at baseline (N=1,837). Symptoms of depression, cannabis abuse, and other psychiatric disorders were assessed with the Diagnostic Interview Schedule. RESULTS: In participants with no baseline depressive symptoms, those with a diagnosis of cannabis abuse at baseline were four times more likely than those with no cannabis abuse diagnosis to have depressive symptoms at the follow up assessment, after adjusting for age, gender, antisocial symptoms, and other baseline covariates. In particular, these participants were more likely to have experienced suicidal ideation and anhedonia during the follow-up period. Among the participants who had no diagnosis of cannabis abuse at baseline, depressive symptoms at baseline failed to significantly predict cannabis abuse at the follow up assessment. CONCLUSIONS: Further research is needed to identify characteristics of individuals who abuse cannabis that account for their higher risk of depression to estimate the degree of impairment resulting from their depression. PMID- 11729022 TI - Long-term outcome of cognitive behavior therapy versus relaxation therapy for chronic fatigue syndrome: a 5-year follow-up study. AB - OBJECTIVE: This study evaluated the long-term outcome of cognitive behavior therapy versus relaxation therapy for patients with chronic fatigue syndrome. METHOD: Sixty patients who participated in a randomized controlled trial of cognitive behavior therapy versus relaxation therapy for chronic fatigue syndrome were invited to complete self-rated measures and participate in a 5-year follow up interview with an assessor who was blind to treatment type. RESULTS: Fifty three patients (88%) participated in the follow-up study: 25 received cognitive behavior therapy and 28 received relaxation therapy. A total of 68% of the patients who received cognitive behavior therapy and 36% who received relaxation therapy rated themselves as "much improved" or "very much improved" at the 5-year follow-up. Significantly more patients receiving cognitive behavior therapy, in relation to those in relaxation therapy, met criteria for complete recovery, were free of relapse, and experienced symptoms that had steadily improved or were consistently mild or absent since treatment ended. Similar proportions were employed, but patients in the cognitive behavior therapy group worked significantly more mean hours per week. Few patients crossed the threshold for "normal" fatigue, despite achieving a good outcome on other measures. Cognitive behavior therapy was positively evaluated and was still used by over 80% of the patients. CONCLUSIONS: Cognitive behavior therapy for chronic fatigue syndrome can produce some lasting benefits but is not a cure. Once therapy ends, some patients have difficulty making further improvements. In the future, attention should be directed toward ensuring that gains are maintained and extended after regular treatment ends. PMID- 11729023 TI - An open-label trial of evidence-based cognitive behavior therapy for nightmares and insomnia in crime victims with PTSD. AB - OBJECTIVE: Insomnia and nightmares are perceived as secondary phenomena in posttraumatic stress disorder (PTSD). Scant treatment research has targeted these two sleep disturbances. This study reports on an open-label trial of cognitive behavior therapy for insomnia and disturbing dreams in crime victims with PTSD. The relationship among nightmares, sleep disturbances, and PTSD symptoms is discussed. METHOD: Sixty-two participants completed a 10-hour group treatment consisting of imagery rehearsal for nightmares and sleep hygiene, stimulus control, and sleep restriction for insomnia. Nightmare frequency, sleep quality, sleep impairment, and ratings for PTSD, anxiety, and depression symptoms were assessed at baseline and at the 3-month follow-up. RESULTS: All measures demonstrated improvement that was roughly equivalent to changes in clinical severity from severe to moderate for sleep quality, sleep impairment, and nightmare frequency, from borderline severe to borderline moderate for PTSD symptoms, and from extremely severe to borderline severe for anxiety and depression symptoms. CONCLUSIONS: In this uncontrolled study, successful treatment for insomnia and nightmares in crime victims was associated with improvement in symptoms of PTSD, anxiety, and depression. Participants with clinical improvements in PTSD symptoms demonstrated significantly greater improvement in sleep quality and nightmare frequency than those whose PTSD symptoms did not improve. PMID- 11729024 TI - Multinuclear magnetic resonance spectroscopy studies of brain purines in major depression. AB - OBJECTIVE: Studies of depressed adults have shown abnormalities in cerebral energy metabolism, as noted by low brain levels of nucleoside triphosphate (NTP), which primarily represents adenosine triphosphate (ATP). This study was undertaken to determine whether proton magnetic resonance spectroscopy (1H MRS) measures of the low-field purine resonance, which arises primarily from adenosine phosphates, can be used to assess abnormalities in cerebral purine metabolism in depressed adults. METHOD: Data from 1H MRS and phosphorus-31 (31P) MRS were acquired for depressed and nondepressed comparison subjects. Intensities of the purine resonance, by 1H MRS (7.5-8.5 ppm), and of NTP, by 31P MRS, were determined. RESULTS: Purine resonance intensities did not differ on average between depressed patients and comparison subjects. However, purine levels were approximately 30% lower in female depressed subjects who subsequently responded to fluoxetine treatment than in those who did not respond. Beta-NTP was lower by 21% in responders than in nonresponders and was correlated with purine levels for the depressed subjects. CONCLUSIONS: Brain purine levels are low in female depressed patients who respond to treatment with fluoxetine, suggesting that response to treatment might be predicted by using 1H MRS. These observations also suggest that agents that increase brain adenosine levels may have antidepressant efficacy. PMID- 11729025 TI - A study of women who stalk. AB - OBJECTIVE: The authors examined whether female stalkers differ from their male counterparts in psychopathology, motivation, behavior, and propensity for violence. METHOD: Female (N=40) and male (N=150) stalkers referred to a forensic mental health clinic were compared. RESULTS: In this cohort, female stalkers were outnumbered by male stalkers by approximately four to one. The demographic characteristics of the groups did not differ, although more male stalkers reported a history of criminal offenses. Higher rates of substance abuse were also noted among the male stalkers, but the psychiatric status of the groups did not otherwise differ. The duration of stalking and the frequency of associated violence were equivalent between groups. The nature of the prior relationship with the victim differed, with female stalkers more likely to target professional contacts and less likely to harass strangers. Female stalkers were also more likely than male stalkers to pursue victims of the same gender. The majority of female stalkers were motivated by the desire to establish intimacy with their victim, whereas men showed a broader range of motivations. CONCLUSIONS: Female and male stalkers vary according to the motivation for their pursuit and their choice of victim. A female stalker typically seeks to attain a close intimacy with her victim, who usually is someone previously known and frequently is a person cast in the professional role of helper. While the contexts for stalking may differ by gender, the intrusiveness of the behaviors and potential for harm does not. PMID- 11729027 TI - Defective relationship between subjective experience and behavior in schizophrenia. AB - OBJECTIVE: The relationship between subjective experience and behavior abnormalities in schizophrenia was investigated. METHOD: Eighteen patients with schizophrenia and 18 normal comparison subjects completed a general knowledge task with two incentive conditions to measure monitoring effectiveness, control sensitivity, and response criterion setting. RESULTS: The patients' levels of monitoring effectiveness and control sensitivity were lower than those of the comparison subjects. The effect of incentives on response criterion values was similar in the two groups. CONCLUSIONS: Patients were impaired in subjectively assessing the correctness of their knowledge, and their behavior was less determined by subjective experience than that of normal subjects. The patients' intact sensitivity to incentives has implications for cognitive remediation. PMID- 11729026 TI - Prevalence of body dysmorphic disorder in a community sample of women. AB - OBJECTIVE: In a large population-based study, the authors examined the prevalence and correlates of body dysmorphic disorder, a debilitating and chronic condition characterized by an imagined defect in appearance. METHOD: Rates and diagnostic correlates of body dysmorphic disorder were examined by using data from the Harvard Study of Moods and Cycles. This study used in-person structured clinical interviews to characterize the diagnostic status of a population-based, cross sectional sample of 318 depressed and 658 nondepressed women between the ages of 36 and 44 who were selected from seven Boston metropolitan area communities. RESULTS: The presence of body dysmorphic disorder was significantly associated with the presence of major depression and anxiety disorders. The authors estimated the overall point prevalence of body dysmorphic disorder as 0.7% in women in this age range in the community. CONCLUSIONS: The authors found that the presence of body dysmorphic disorder was linked to the presence of major depression and anxiety disorders, which is similar to findings in clinical studies. Their estimate of the point prevalence of body dysmorphic disorder is consistent with data from a community-based sample of Italian women and suggests a prevalence similar to that of other serious psychiatric disorders in women (e.g., schizophrenia and drug abuse and dependence). These prevalence data encourage the further development of treatment options for this debilitating condition. PMID- 11729028 TI - Measuring P50 suppression and prepulse inhibition in a single recording session. AB - OBJECTIVE: Two distinct measures have been used to assess inhibitory gating deficits in schizophrenia patients: P50 suppression and prepulse inhibition of the startle response. It remains unclear whether both measures can be assessed in a single testing session. METHOD: Twelve normal subjects underwent testing in a carefully designed combined P50/prepulse inhibition session using stimulus characteristics similar to those described in the existing literature. RESULTS: The levels of both P50 suppression and prepulse inhibition obtained in the combined session were highly similar to those obtained in independent testing of previous cohorts of normal subjects. As in previous experiments, P50 suppression and prepulse inhibition were not significantly correlated. CONCLUSIONS: Measuring P50 suppression and prepulse inhibition in a single session is feasible and offers a unique opportunity to assess these two distinct gating measures contemporaneously in cohorts of normal comparison subjects and schizophrenia patients, so that temporal shifts in one or both measures are minimized. PMID- 11729029 TI - Neurophysiological evidence of corollary discharge dysfunction in schizophrenia. AB - OBJECTIVE: Speaking is hypothesized to generate a corollary discharge of motor speech commands transmitted to the auditory cortex, dampening its response to self-generated speech sounds. Event-related potentials were used to test whether failures of corollary discharge during speech contribute to the pathophysiology of schizophrenia. METHOD: The N1 component of the event-related potential elicited by vowels was recorded while the vowels were spoken by seven patients with schizophrenia and seven healthy comparison subjects and while the same vowels were played back. RESULTS: In the healthy subjects, the N1 elicited by spoken vowels was smaller than the N1 elicited by played-back vowels. This reduction in N1 elicited by spoken vowels was not observed in the patients with schizophrenia. CONCLUSIONS: These findings provide direct neurophysiological evidence for a corollary discharge that dampens sensory responses to self generated, relative to externally presented, percepts in healthy comparison subjects and its failure in patients with schizophrenia. PMID- 11729030 TI - A placebo-controlled trial of omega-3 fatty acid (ethyl eicosapentaenoic acid) supplementation for residual symptoms and cognitive impairment in schizophrenia. AB - OBJECTIVE: This study determined if augmentation of neuroleptics with 3 g/day of ethyl eicosapentaenoic acid (EPA) improves symptoms and cognition in patients with schizophrenia or schizoaffective disorder. METHOD: Eighty-seven patients meeting criteria for schizophrenia or schizoaffective disorder who had residual symptoms despite neuroleptic treatment were randomly assigned to receive either 3 g/day of ethyl EPA (N=43) or placebo (N=44) in a 16-week, double-blind supplementation trial. Assessments were performed at baseline and at weeks 1, 2, 4, 8, 12, and 16; a cognitive battery was administered at baseline and at week 16. RESULTS: No differences were found between groups in positive or negative symptoms, mood, cognition, or global impression ratings. Results were similar for the intention-to-treat (N=87) and completer (N=75) groups. CONCLUSIONS: For schizophrenia patients treated with 3 g/day of ethyl EPA, improvement in residual symptoms and cognitive impairment was no greater than for schizophrenia patients treated with placebo. PMID- 11729031 TI - Influence of serotonin-transporter-linked promoter region polymorphism on platelet activation in geriatric depression. AB - OBJECTIVE: Depression has been associated with increased platelet activation. Variations in the serotonin-transporter-linked promoter region (5-HTTLPR) polymorphism may influence the degree of activation. The authors examined the association among depression, platelet activation, and 5-HTTLPR genotype. METHOD: Elderly subjects with (N=61) and without (N=12) major depression were assessed for cognitive impairment, cardiovascular disease, and two indices of platelet activation. The depressed subjects were genotyped for the 5-HTTLPR polymorphism. RESULTS: The depressed subjects were older, were more cognitively impaired, and had higher platelet factor 4 and beta-thromboglobulin levels; cardiovascular disease was minimal in both groups. In the depressed group, subjects with the 5 HTTLPR l/l genotype had significantly higher platelet factor 4 and beta thromboglobulin levels. CONCLUSIONS: Platelet activation is increased in elderly depressed patients, especially those with the 5-HTTLPR l/l genotype. This finding suggests how genetic differences may influence cardiovascular mortality in depressed patients with ischemic heart disease. PMID- 11729032 TI - Antidepressant treatment of depression in the Finnish general population. AB - OBJECTIVE: Antidepressant use has increased in the last decade, but whether depression continues to be undertreated is unknown. The authors investigated the prevalence of antidepressant treatment and its predictors in a recent general population sample of depressed subjects. METHOD: As part of the Finnish Health Care Survey, in 1996 a representative sample of Finns (N=5,993) aged 15-75 years underwent a standardized face-to-face interview that used the DSM-III-R criteria for major depressive episode. RESULTS: Only 13% of subjects with a major depressive episode during the preceding 12 months (70 of 557) reported current use of an antidepressant. In logistic regression models, use of psychiatric services for depression, regular use of any other medication, more than 1 month of sick leave, and smoking were associated with antidepressant treatment. CONCLUSIONS: Most depressed subjects in 1996 in Finland were not receiving antidepressant treatment despite the several-fold increase in antidepressant use in the 1990s. PMID- 11729033 TI - Pindolol augmentation of selective serotonin reuptake inhibitors: PET evidence that the dose used in clinical trials is too low. AB - OBJECTIVE: Positron emission tomography (PET) was used to examine whether the dose of pindolol used to augment antidepressant medication achieves a significant occupancy of the serotonin type 1A (5-HT(1A)) autoreceptor in depressed patients receiving medication. METHOD: The authors examined eight depressed patients on one of two regimes of pindolol (2.5 mg t.i.d. and 5.0 mg t.i.d.) with PET and [11C]WAY-100635. RESULTS: The 5-mg t.i.d. regime achieved a modest (19%) but significant occupancy of the 5-HT(1A) autoreceptor, while the regime used in the vast majority of clinical trials (2.5 mg t.i.d.) did not achieve a significant occupancy. CONCLUSIONS: The dose of pindolol used in clinical trials is suboptimal and may explain the inconsistent results. Therefore, a thorough test of pindolol's efficacy will necessitate doses higher than those used in present clinical trials. PMID- 11729035 TI - Bright light therapy for schizoaffective disorder. PMID- 11729034 TI - Cortisol feedback during the HPA quiescent period in patients with major depression. AB - OBJECTIVE: The authors tested the hypothesis that patients with major depression have a defect in the mechanism by which cortisol exerts negative feedback on the hypothalamic-pituitary-adrenal (HPA) axis during the HPA axis quiescent period. METHOD: Twenty-nine patients with major depression and 25 healthy comparison subjects were randomly assigned to administration of 15 mg cortisol or placebo infused over 2 hours beginning at 7:00 p.m. Cortisol and ACTH levels were measured at baseline and every 30 minutes from 7:30 p.m. to 11:00 p.m. RESULTS: Differences between the patients and the comparison subjects in the ACTH response to the cortisol infusion, relative to the ACTH response to placebo, were not found. CONCLUSIONS: The results provide some evidence that patients with major depression do not have an abnormality of cortisol feedback during the HPA axis quiescent period. PMID- 11729036 TI - Imipramine toxicity and terbinafine. PMID- 11729037 TI - Olanzapine-induced priapism. PMID- 11729038 TI - Continuation of clozapine after priapism. PMID- 11729039 TI - Venlafaxine- and trazodone-induced serotonin syndrome. PMID- 11729040 TI - Venlafaxine and breast-feeding. PMID- 11729041 TI - ECT for psychotic depression associated with a brain tumor. PMID- 11729042 TI - End-of-life care content in textbooks. PMID- 11729043 TI - Psychiatric pharmacists. PMID- 11729044 TI - Psychotherapy by nonphysicians. PMID- 11729045 TI - Tribute to Lasegue. PMID- 11729047 TI - Psychosocial treatment for schizophrenia. PMID- 11729049 TI - Research on women. PMID- 11729051 TI - Drugs on the internet. PMID- 11729053 TI - Adolescents with schizophrenia. PMID- 11729067 TI - Ross River virus: ecology and distribution. AB - Ross River virus is the most common mosquito-borne pathogen in Australia, and approximately 5000 human cases are reported annually. The infection is not fatal, but there is considerable morbidity associated with a debilitating polyarthritis that is the major symptom. The virus is annually active in most regions of Australia, but exists as strains that vary in virulence. Native macropods are thought to be the natural vertebrate hosts, although horses and humans may be involved during epidemic activity, and vertical transmission of the virus occurs in mosquitoes. Different mosquito species are involved as vectors in various regions and in different seasonal and environmental conditions. In coastal areas the saltmarsh mosquitoes Aedes camptorhynchus and Ae. vigilax are the most important vectors in southern and northern regions, respectively, whereas in inland areas Culex annulirostris is the most important vector, although various Aedes species can be involved depending on region and conditions, and the epidemiology of the disease and vector control imperatives vary with circumstance concomitantly. PMID- 11729068 TI - Biology and management of the smokybrown cockroach. AB - The smokybrown cockroach, Periplaneta fuliginosa (Serville), has become an increasingly important peridomestic pest throughout much of the southeastern United States, Japan, and southeast Asia. We provide a brief description of the species and clarify its worldwide distribution. Aspects of smokybrown cockroach behavior, biology, ecology, and physiology are discussed in the context of this species' successful exploitation of the urban environment. We describe smokybrown cockroach management based upon a model that relates house and landscape characteristics to cockroach population size. Finally, we illustrate the effectiveness of a model-based IPM system that increases control while reducing use of insecticides. PMID- 11729069 TI - Sequestration of defensive substances from plants by Lepidoptera. AB - A number of aposematic butterfly and diurnal moth species sequester unpalatable or toxic substances from their host plants rather than manufacturing their own defensive substances. Despite a great diversity in their life histories, there are some general features in the selective utilization of plant secondary metabolites to achieve effective protection from predators. This review illustrates the biochemical, physiological, and ecological characteristics of phytochemical-based defense systems that can shed light on the evolution of the widely developed sequestering lifestyles among the Lepidoptera. PMID- 11729070 TI - Regulation of diapause. AB - Environmental and hormonal regulators of diapause have been reasonably well defined, but our understanding of the molecular regulation of diapause remains in its infancy. Though many genes are shut down during diapause, others are specifically expressed at this time. Classes of diapause-upregulated genes can be distinguished based on their expression patterns: Some are upregulated throughout diapause, and others are expressed only in early diapause, late diapause, or intermittently throughout diapause. The termination of diapause is accompanied by a rapid decline in expression of the diapause-upregulated genes and, conversely, an elevation in expression of many genes that were downregulated during diapause. A comparison of insect diapause with other forms of dormancy in plants and animals suggests that upregulation of a subset of heat shock protein genes may be one feature common to different types of dormancies. PMID- 11729071 TI - Bacterial symbionts of the triatominae and their potential use in control of Chagas disease transmission. AB - Chagas disease is caused by the parasitic protozoan Trypanosoma cruzi and transmitted by insects in the family Reduviidae, subfamily Triatominae, commonly known as kissing bugs. Because these insects feed throughout their entire developmental cycle on vertebrate blood, they harbor populations of symbiotic bacteria in their intestinal track that produce nutrients that are lacking in the insects' limited diet. It is possible to cultivate these bacteria, genetically modify them, and place them back into their insect host, thus generating a paratransgenic insect. This procedure has allowed the expression of antitrypanosomal gene products in the insect gut, thereby resulting in insects that are incapable of transmitting Chagas disease. A method has been developed that would allow introduction and spread of genetically modified symbionts into natural populations of kissing bugs, thus leading potentially to a transgenic intervention tool for use as a part of an integrated vector control approach. PMID- 11729072 TI - Strategies and statistics of sampling for rare individuals. AB - Diverse subdisciplines within entomology recognize the detection of rare individuals as the precursor to effective management of these individuals. Unfortunately, detection methods have often developed on a case-by-case basis, and advances in one subdiscipline have not carried over to similarly related fields. The biology of a particular organism will certainly affect sampling methods, but the underlying principles governing the power of a sampling strategy to detect rare individuals will apply across taxa. Our review of the sampling literature demonstrates the common problem of detecting rare individuals, reviews the fundamentals of probability theory as a foundation for any monitoring program, and discusses the inferences that can be drawn from samples, especially when resources limit sampling efforts. Particular emphasis is placed on binomial , beta-binomial-, and hypergeometric-based sampling strategies as they pertain to quarantine inspections for exotic pests, veterinary/medical entomology, and insecticide resistance monitoring. PMID- 11729073 TI - Biology and management of the Japanese beetle. AB - The Japanese beetle, Popillia japonica Newman, an introduced scarab, has become the most widespread and destructive insect pest of turf, landscapes, and nursery crops in the eastern United States. It also damages many fruit, garden, and field crops. This review emphasizes recent research on the beetle's biology and management. Adults feed on leaves, flowers, or fruits of more than 300 plant species. Adaptations mediating their host finding, dietary range, mating, and oviposition are discussed. We also address abiotic and biotic factors affecting population dynamics of the root-feeding larvae. Japanese beetle grubs are widely controlled with preventive soil insecticides, but options for remedial control of adults and larvae presently are limited. Advances in understanding host plant resistance, entomopathogens, and other biorational approaches may provide more options for integrated management. Despite ongoing regulatory efforts, the Japanese beetle remains a threat as an invasive species. PMID- 11729074 TI - Biology and ecology of higher Diptera from freshwater wetlands. AB - Although studies of freshwater entomofauna frequently do not include the biodiversity and ecological roles of higher Diptera, cyclorraphous flies are often numerous and species rich in wetlands. Seventeen families are commonly found in freshwater wetlands, with Ephydridae, Chloropidae, Sciomyzidae, Sphaeroceridae, and Scathophagidae being among the most important in terms of population size and species richness. Difficulty with sampling cryptic larval habitats and species identification challenges may account for the exclusion of acalyptrate and other dipterans from wetlands ecology studies. Large populations are facilitated by the high productivity of freshwater wetlands and the high intrinsic rate of increase characteristic of many species. Higher dipterans exist in all freshwater wetland types, are microhabitat selective, and play significant roles in food webs. The varied strategies for food acquisition and patterns of spatial and temporal distribution limit ecological overlap among the higher Diptera. PMID- 11729075 TI - Invasions by insect vectors of human disease. AB - Nonindigenous vectors that arrive, establish, and spread in new areas have fomented throughout recorded history epidemics of human diseases such as malaria, yellow fever, typhus, and plague. Although some vagile vectors, such as adults of black flies, biting midges, and tsetse flies, have dispersed into new habitats by flight or wind, human-aided transport is responsible for the arrival and spread of most invasive vectors, such as anthropophilic fleas, lice, kissing bugs, and mosquitoes. From the fifteenth century to the present, successive waves of invasion of the vector mosquitoes Aedes aegypti, the Culex pipiens Complex, and, most recently, Aedes albopictus have been facilitated by worldwide ship transport. Aircraft have been comparatively unimportant for the transport of mosquito invaders. Mosquito species that occupy transportable container habitats, such as water-holding automobile tires, have been especially successful as recent invaders. Propagule pressure, previous success, and adaptations to human habits appear to favor successful invasions by vectors. PMID- 11729076 TI - Omnivory in terrestrial arthropods: mixing plant and prey diets. AB - Many terrestrial communities include omnivorous arthropods that feed on both prey and plant resources. In this review we first discuss some unique morphological, physiological, and behavioral traits that enable omnivores to exploit such dissimilar foods, and we explore possible evolutionary pathways to omnivory. We then examine possible benefits and costs of omnivory, describe the relationships between omnivory and other high-order complex trophic interactions, and consider the stability level of communities with closed-loop omnivory. Finally, we explore some of the implications of omnivory for crop damage and for biological, chemical, and cultural control practices. We conclude that the growing realization of the ubiquity of omnivory in nature may require a change in our view of the structure and function of ecological systems. PMID- 11729077 TI - How to be a fig wasp. AB - In the two decades since Janzen described how to be a fig, more than 200 papers have appeared on fig wasps (Agaonidae) and their host plants (Ficus spp., Moraceae). Fig pollination is now widely regarded as a model system for the study of coevolved mutualism, and earlier reviews have focused on the evolution of resource conflicts between pollinating fig wasps, their hosts, and their parasites. Fig wasps have also been a focus of research on sex ratio evolution, the evolution of virulence, coevolution, population genetics, host-parasitoid interactions, community ecology, historical biogeography, and conservation biology. This new synthesis of fig wasp research attempts to integrate recent contributions with the older literature and to promote research on diverse topics ranging from behavioral ecology to molecular evolution. PMID- 11729078 TI - Alternatives to methyl bromide treatments for stored-product and quarantine insects. AB - Methyl bromide is used to control insects as a space fumigant in flour and feed mills and ship holds, as a product fumigant for some fruit and cereals, and for general quarantine purposes. Methyl bromide acts rapidly, controlling insects in less than 48 h in space fumigations, and it has a wide spectrum of activity, controlling not only insects but also nematodes and plant-pathogenic microbes. This chemical will be banned in 2005 in developed countries, except for exceptional quarantine purposes, because it depletes ozone in the atmosphere. Many alternatives have been tested as replacements for methyl bromide, from physical control methods such as heat, cold, and sanitation to fumigant replacements such as phosphine, sulfuryl fluoride, and carbonyl sulfide, among others. Individual situations will require their own type of pest control techniques, but the most promising include integrated pest management tactics and combinations of treatments such as phosphine, carbon dioxide, and heat. PMID- 11729079 TI - Ecology and behavior of first instar larval Lepidoptera. AB - Neonate Lepidoptera are confronted with the daunting task of establishing themselves on a food plant. The factors relevant to this process need to be considered at spatial and temporal scales relevant to the larva and not the investigator. Neonates have to cope with an array of plant surface characters as well as internal characters once the integument is ruptured. These characters, as well as microclimatic conditions, vary within and between plant modules and interact with larval feeding requirements, strongly affecting movement behavior, which may be extensive even for such small organisms. In addition to these factors, there is an array of predators, pathogens, and parasitoids with which first instars must contend. Not surprisingly, mortality in neonates is high but can vary widely. Experimental and manipulative studies, as well as detailed observations of the animal, are vital if the subtle interaction of factors responsible for this high and variable mortality are to be understood. These studies are essential for an understanding of theories linking female oviposition behavior with larval survival, plant defense theory, and population dynamics, as well as modern crop resistance breeding programs. PMID- 11729080 TI - Arthropod allergens and human health. AB - Many species of arthropods are the sources of potent allergens that sensitize and induce IgE-mediated allergic reactions in humans. Most of these arthropod allergens are proteins, and the allergic response mechanism to these allergens is the same as it is for allergens from other sources such as plant pollens, molds, and foods. Aside from ingestion of crustaceans (shrimp, lobster), among arthropods, humans have the greatest contact with insects and mites, and as a result allergies to these two groups of arthropods have been the most frequently reported. Because of the large number of people affected by allergic reactions to stinging insects, cockroaches, and dust mites, many allergens of these organisms have been extensively studied, purified, and immunobiochemically characterized and for some recombinant allergens, produced. Cocktails of these recombinant allergens have the potential for use in diagnosis and immunotherapy. In this chapter, we review the insects and mites that induce allergic reactions. Where the information exists, the immunobiochemical characterization of the allergens and the frequency of sensitivity or clinical reactivity in the human population are also reviewed. As background, the beginning of this review includes sections that define allergens, explain the allergic reaction mechanism, and describe the methods for naming allergens. PMID- 11729081 TI - Competitive displacement among insects and arachnids. AB - Competitive displacement is the most severe outcome of interspecific competition. For the purposes of this review, we define this type of displacement as the removal of a formerly established species from a habitat as a result of direct or indirect competitive interactions with another species. We reviewed the literature for recent putative cases of competitive displacement among insects and arachnids and assessed the evidence for the role of interspecific competition in these displacements. We found evidence for mechanisms of both exploitation and interference competition operating in these cases of competitive displacement. Many of the cases that we identified involve the operation of more than one competitive mechanism, and many cases were mediated by other noncompetitive factors. Most, but not all, of these displacements occurred between closely related species. In the majority of cases, exotic species displaced native species or previously established exotic species, often in anthropogenically altered habitats. The cases that we identified have occurred across a broad range of taxa and environments. Therefore we suggest that competitive displacement has the potential to be a widespread phenomenon, and the frequency of these displacement events may increase, given the ever-increasing degree of anthropogenic changes to the environment. A greater awareness of competitive displacement events should lead to more studies documenting the relative importance of key factors and developing hypotheses that explain observed patterns. PMID- 11729082 TI - Endocrine insights into the evolution of metamorphosis in insects. AB - This review explores the roles of ecdysone and juvenile hormone (JH) in the evolution of complete metamorphosis and how metamorphosis, in turn, has impacted endocrine signaling. JH is a key player in the evolution of metamorphosis because it can act on embryos from more basal insect groups to suppress morphogenesis and cause premature differentiation, functions needed for transforming the transitional pronymphal stage of hemimetabolous insects into a functional larval stage. In the ancestral condition, imaginal-related growth is then delayed until JH finally disappears during the last larval instar. In the more derived groups of the Holometabola, selective tissues have escaped this JH suppression to form early-growing imaginal discs. We discuss how complete metamorphosis may have influenced the molecular aspects of both ecdysone and JH signaling. PMID- 11729083 TI - Biochemistry and genetics of insect resistance to Bacillus thuringiensis. AB - Bacillus thuringiensis (Bt) is a valuable source of insecticidal proteins for use in conventional sprayable formulations and in transgenic crops, and it is the most promising alternative to synthetic insecticides. However, evolution of resistance in insect populations is a serious threat to this technology. So far, only one insect species has evolved significant levels of resistance in the field, but laboratory selection experiments have shown the high potential of other species to evolve resistance against Bt. We have reviewed the current knowledge on the biochemical mechanisms and genetics of resistance to Bt products and insecticidal crystal proteins. The understanding of the biochemical and genetic basis of resistance to Bt can help design appropriate management tactics to delay or reduce the evolution of resistance in insect populations. PMID- 11729084 TI - Iron metabolism in insects. AB - Like other organisms, insects must balance two properties of ionic iron, that of an essential nutrient and a potent toxin. Iron must be acquired to provide catalysis for oxidative metabolism, but it must be controlled to avoid destructive oxidative reactions. Insects have evolved distinctive forms of the serum iron transport protein, transferrin, and the storage protein, ferritin. These proteins may serve different functions in insects than they do in other organisms. A form of translational control of protein synthesis by iron in insects is similar to that of vertebrates. The Drosophila melanogaster genome contains many genes that may encode other proteins involved in iron metabolism. PMID- 11729085 TI - Can generalist predators be effective biocontrol agents? AB - Theoretical developments are helping us to comprehend the basic parameters governing the dynamics of the interactions between generalist predators and their many pest and nonpest prey. In practice, however, inter- and intraspecific interactions between generalist predators, and between the predators and their prey, within multispecies systems under the influence of rapidly changing biotic and abiotic variables are difficult to predict. We discuss trade-offs between the relative merits of specialists and generalists that allow both to be effective, and often complementary, under different circumstances. A review of manipulative field studies showed that in approximately 75% of cases, generalist predators, whether single species or species assemblages, reduced pest numbers significantly. Techniques for manipulating predator numbers to enhance pest control at different scales are discussed. We now need to find ways of disentangling the factors influencing positive and negative interactions within natural enemy communities in order to optimize beneficial synergies leading to pest control. PMID- 11729086 TI - Arthropods on islands: colonization, speciation, and conservation. AB - Islands have traditionally been considered to be any relatively small body of land completely surrounded by water. However, their primary biological characteristic, an extended period of isolation from a source of colonists, is common also to many situations on continents. Accordingly, theories and predictions developed for true islands have been applied to a huge array of systems, from rock pools, to single tree species in forests, to oceanic islands. Here, we examine the literature on islands in the broadest sense (i.e., whether surrounded by water or any other uninhabitable matrix) as it pertains to terrestrial arthropods. We categorize islands according to the features they share. The primary distinction between different island systems is "darwinian" islands (formed de novo) and "fragment" islands. In the former, the islands have never been in contact with the source of colonists and have abundant "empty" ecological niche space. On these islands, species numbers will initially increase through immigration, the rate depending on the degree of isolation. If isolation persists, over time species formation will result in "neo-endemics." When isolation is extreme, the ecological space will gradually be filled through speciation (rather than immigration) and adaptive radiation of neo-endemics. Fragment islands are fundamentally different. In these islands, the ecological space will initially be filled as a consequence of connection to the source of colonists prior to insularization. Species numbers will decrease following fragmentation through the process of relaxation. If these islands become more isolated, species will eventually arise through relictualization with the formation of "paleo-endemics." Given sufficient time, this process can result in generic level endemism on ancient fragment islands, a phenomenon well illustrated in Madagascar and New Zealand. Recognizing the distinction between the different kinds of islands is fundamental for understanding emerging patterns on each, in particular speciation, biodiversity (e.g., neo-endemics versus paleo-endemics), and conservation (e.g., naivete in interactions with alien species). PMID- 11729087 TI - The population biology of oak gall wasps (Hymenoptera: Cynipidae). AB - Oak gall wasps (Hymenoptera: Cynipidae, Cynipini) are characterized by possession of complex cyclically parthenogenetic life cycles and the ability to induce a wide diversity of highly complex species- and generation-specific galls on oaks and other Fagaceae. The galls support species-rich, closed communities of inquilines and parasitoids that have become a model system in community ecology. We review recent advances in the ecology of oak cynipids, with particular emphasis on life cycle characteristics and the dynamics of the interactions between host plants, gall wasps, and natural enemies. We assess the importance of gall traits in structuring oak cynipid communities and summarize the evidence for bottom-up and top-down effects across trophic levels. We identify major unanswered questions and suggest approaches for the future. PMID- 11729088 TI - Short, long, and beyond: molecular and embryological approaches to insect segmentation. AB - Over the past dozen years, studies comparing the expression of orthologues of the Drosophila segmentation genes among various insects have served to broaden our view of the ways in which insects make segments. The molecular data suggest that, although the overall genetic mechanisms of segmentation during embryogenesis have been conserved, the details of this process vary both within and between various insect orders. Here we summarize comparative gene expression data relevant to segmentation with an emphasis on understanding the extent of molecular patterning prior to gastrulation. These results are discussed in embryological context with an eye toward understanding the evolution of segmentation within insects. PMID- 11729089 TI - Biology and management of economically important lepidopteran cereal stem borers in Africa. AB - Cereals (maize, sorghum, millet, rice) are extremely important crops grown in Africa for human consumption. Of the various insect pests attacking cereal crops in Africa, lepidopteran stem borers are by far the most injurious. All 21 economically important stem borers of cultivated grasses in Africa are indigenous except Chilo partellus, which invaded the continent from India, and C. sacchariphagus, which has recently been found in sugarcane in Mozambique. C. partellus is competitively displacing indigenous stem borers in East and southern Africa. A parasitoid, Cotesia flavipes, was introduced from Pakistan for biological control of C. partellus and caused a 32-55% decrease in stem borer densities. This article is an attempt to summarize the status of knowledge about economically important cereal stem borers in Africa with emphasis on their distribution, pest status and yield losses, diapause, natural enemies, cultural control, host plant resistance, and biological control. Special attention is given to Busseola fusca and C. partellus, the most important pests of maize and grain sorghum. PMID- 11729090 TI - The ecology and evolution of ant association in the Lycaenidae (Lepidoptera). AB - The estimated 6000 species of Lycaenidae account for about one third of all Papilionoidea. The majority of lycaenids have associations with ants that can be facultative or obligate and range from mutualism to parasitism. Lycaenid larvae and pupae employ complex chemical and acoustical signals to manipulate ants. Cost/benefit analyses have demonstrated multiple trade-offs involved in myrmecophily. Both demographic and phylogenetic evidence indicate that ant association has shaped the evolution of obligately associated groups. Parasitism typically arises from mutualism with ants, and entomophagous species are disproportionately common in the Lycaenidae compared with other Lepidoptera. Obligate associations are more common in the Southern Hemisphere, in part because highly ant-associated lineages make up a larger proportion of the fauna in these regions. Further research on phylogeny and natural history, particularly of the Neotropical fauna, will be necessary to understand the role ant association has played in the evolution of the Lycaenidae. PMID- 11729091 TI - Sympatric speciation in phytophagous insects: moving beyond controversy? AB - Sympatric speciation is the splitting of one evolutionary lineage into two without the occurrence of geographic isolation. The concept has been intimately tied to entomology since the 1860s, when Benjamin Walsh proposed that many host specific phytophagous insects originate by shifting and adapting to new host plant species. If true, sympatric speciation would have tremendous implications for our understanding of species and their origins, biodiversity (25-40% of all animals are thought to be phytophagous specialists), insect-plant coevolution, community ecology, phylogenetics, and systematics, as well as practical significance for the management of insect pests. During much of the twentieth century sympatric speciation was viewed as much less plausible than geographic (allopatric) speciation. However, empirical field studies, laboratory experiments, developments in population genetics theory, and phylogenetic and biogeographic data have all recently combined to shed a more favorable light on the process. We review the evidence for sympatric speciation via host shifting for phytophagous insects and propose a set of testable predictions for distinguishing geographic mode (allopatric versus sympatric) of divergence. Our conclusion is that sympatric speciation is a viable hypothesis. We highlight areas where more thorough testing is needed to move sympatric speciation into the realm of accepted scientific theory. PMID- 11729092 TI - Host plant quality and fecundity in herbivorous insects. AB - Host plant quality is a key determinant of the fecundity of herbivorous insects. Components of host plant quality (such as carbon, nitrogen, and defensive metabolites) directly affect potential and achieved herbivore fecundity. The responses of insect herbivores to changes in host plant quality vary within and between feeding guilds. Host plant quality also affects insect reproductive strategies: Egg size and quality, the allocation of resources to eggs, and the choice of oviposition sites may all be influenced by plant quality, as may egg or embryo resorption on poor-quality hosts. Many insect herbivores change the quality of their host plants, affecting both inter- and intraspecific interactions. Higher-trophic level interactions, such as the performance of predators and parasitoids, may also be affected by host plant quality. We conclude that host plant quality affects the fecundity of herbivorous insects at both the individual and the population scale. PMID- 11729093 TI - Economic, ecological, food safety, and social consequences of the deployment of bt transgenic plants. AB - Transgenic plants expressing insecticidal proteins from the bacterium, Bacillus thuringiensis (Bt), are revolutionizing agriculture. Bt, which had limited use as a foliar insecticide, has become a major insecticide because genes that produce Bt toxins have been engineered into major crops grown on 11.4 million ha worldwide in 2000. Based on the data collected to date, generally these crops have shown positive economic benefits to growers and reduced the use of other insecticides. The potential ecological and human health consequences of Bt plants, including effects on nontarget organisms, food safety, and the development of resistant insect populations, are being compared for Bt plants and alternative insect management strategies. Scientists do not have full knowledge of the risks and benefits of any insect management strategies. Bt plants were deployed with the expectation that the risks would be lower than current or alternative technologies and that the benefits would be greater. Based on the data to date, these expectations seem valid. PMID- 11729094 TI - Control and biochemical nature of the ecdysteroidogenic pathway. AB - Molting is elicited by a critical titer of ecdysteroids that includes the principal molting hormone, 20-hydroxyecdysone (20E), and ecdysone (E), which is the precursor of 20E but also has morphogenetic roles of its own. The prothoracic glands are the predominate source of ecdysteroids, and the rate of synthesis of these polyhydroxylated sterols is critical for molting and metamorphosis. This review concerns three aspects of ecdysteroidogenesis: (a) how the brain neuropeptide prothoracicotropic hormone (PTTH) initiates a transductory cascade in cells of the prothoracic gland, which results in an increased rate of ecdysteroid biosynthesis (upregulation); (b) how the concentrations of 20E in the hemolymph feed back on the prothoracic gland to decrease rates of ecdysteroidogenesis (downregulation); and (c) how the prothoracic gland cells convert cholesterol to the precursor of E and then 20E, a series of reactions only now being understood because of the use of a combination of classical biochemistry and molecular genetics. PMID- 11729095 TI - The biology of the dance language. AB - Honey bee foragers dance to communicate the spatial location of food and other resources to their nestmates. This remarkable communication system has long served as an important model system for studying mechanisms and evolution of complex behavior. I provide a broad synthesis of recent research on dance communication, concentrating on the areas that are currently the focus of active research. Specific issues considered are as follows: (a) the sensory and integrative mechanisms underlying the processing of spatial information in dance communication, (b) the role of dance communication in regulating the recruitment of workers to resources in the environment, (c) the evolution of the dance language, and (d) the adaptive fine-tuning of the dance for efficient spatial communication. PMID- 11729097 TI - In vivo targeting of SF/HGF and c-met expression via U1snRNA/ribozymes inhibits glioma growth and angiogenesis and promotes apoptosis. AB - The multifunctional growth factor scatter factor/hepatocyte growth factor (SF/HGF) and its receptor c-met have been implicated in the genesis, malignant progression, and chemo/radioresistance of multiple human malignancies, including gliomas. We examined the antitumor effects of targeting SF/HGF and c-met expression in pre-established glioma xenografts by using novel chimeric U1snRNA/ribozymes. Transient expression of anti-SF/HGF and anti-c-met U1snRNA/ribozymes inhibited SF/HGF and c-met expression, c-met receptor activation, tumor cell migration, and anchorage-independent colony formation in vitro. Delivery of U1snRNA/ribozymes to established subcutaneous glioma xenografts via liposome-DNA complexes significantly inhibited tumor growth as well as tumor SF/HGF and c-met expression levels. Histologic analysis of tumors treated with U1snRNA/ribozymes showed a significant decrease in blood vessel density, an increase in activation of the pro-apoptotic enzyme caspase-3, and an increase in tumor cell apoptosis. Treatment of animals bearing intracranial glioma xenografts with anti-SF/HGF and anti-c-met U1snRNA/ribozymes by either intratumoral injections of adenoviruses expressing the transgenes or intravenous injections of U1snRNA/ribozyme-liposome complexes substantially inhibited tumor growth and promoted animal survival. We demonstrate that SF/HGF and/or c-met expression can be targeted in vivo to inhibit tumor growth. In addition, our findings represent the first in vivo application of chimeric U1snRNA/ribozymes, which have numerous potential therapeutic gene-targeting applications. PMID- 11729098 TI - Creatine kinase is physically associated with the cardiac ATP-sensitive K+ channel in vivo. AB - Cardiac sarcolemmal ATP-sensitive K+ (KATP) channels, composed of Kir6.2 and SUR2A subunits, couple the metabolic status of cells with the membrane excitability. Based on previous functional studies, we have hypothesized that creatine kinase (CK) may be a part of the sarcolemmal KATP channel protein complex. The inside-out and whole cell patch clamp electrophysiology applied on guinea pig cardiomyocytes showed that substrates of CK regulate KATP channels activity. Following immunoprecipitation of guinea-pig cardiac membrane fraction with the anti-SUR2 antibody, Coomassie blue staining revealed, besides Kir6.2 and SUR2A, a polypeptide at approximately 48 kDa. Western blotting analysis confirmed the nature of putative Kir6.2 and SUR2A, whereas matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis identified p48 kDa as a muscle form of CK. In addition, the CK activity was found in the anti SUR2A immunoprecipitate and the cross reactivity between an anti-CK antibody and the anti-SUR2A immunoprecipitate was observed as well as vice verse. Further results obtained at the level of recombinant channel subunits demonstrated that CK is directly physically associated with the SUR2A, but not the Kir6.2, subunit. All together, these results suggest that the CK is associated with SUR2A subunit in vivo, which is an integral part of the sarcolemmal KATP channel protein complex. PMID- 11729099 TI - A novel approach to induce cell cycle reentry in terminally differentiated muscle cells. AB - During terminal differentiation, skeletal muscle cells permanently retract from the cell cycle. We and others have shown previously that this cell cycle withdrawal is an actively maintained state that can be reversed by transient expression of the SV40 large T antigen. In an attempt to avoid the hazards of gene transfer and the difficulties of regulating transgene expression, we have now used this cellular system as a model to test whether direct protein delivery could constitute a feasible alternative or complementing strategy to gene therapy based approaches. Taking advantage of the recently described intercellular trafficking properties of the herpes simplex virus I VP22 protein, we have constructed a chimeric VP22-SV40 large T antigen fusion protein and shown that it can spread into terminally differentiated myotubes where it accumulates in the nucleus. This fusion protein retains the ability to override the cell cycle arrest as shown for SV40 large T antigen alone. Our results clearly show that the transduced fusion protein remains capable of inducing S-phase and mitosis in these otherwise terminally differentiated cells and opens now the way to exploit this novel strategy for tissue regeneration. PMID- 11729100 TI - Selective microvascular dysfunction in mice lacking the gene encoding for desmin. AB - The intermediate filament desmin has a key role in the integrity and contractility of skeletal and cardiac myocytes. Its absence or aggregation leads to cardiomyopathies. In arteries desmin is distributed heterogeneously; vascular disorders might also occur in its absence. We studied endothelial and muscular functions in arteries from mice lacking desmin (des-/-), compared with control (des+/+). Carotid and mesenteric resistance arteries were mounted in vitro in arteriographs. Desmin was located exclusively in smooth muscle cells. In arteries from des-/- mice, pressure-induced (myogenic) tone was unchanged, but agonist induced tone decreased in resistance arteries (no change in large arteries). Flow (shear stress)- and acetylcholine-induced, endothelium-dependent dilation, as well as endothelium-independent dilation, were also decreased in resistance arteries. To our knowledge, this is the first study of vascular contractile and dilatory functions in arteries lacking desmin. Although vascular reactivity was normal in large arteries, it decreased strongly in small resistance arteries. Thus, desmin is required in vascular smooth muscle cells and in resistance arteries, for efficient control of vascular tone and consequently for an optimal blood flow supply. This microvascular defect found in the absence of desmin might play a major role in myopathies seen in desmin-related diseases. PMID- 11729101 TI - Influence of alpha-melanocyte-stimulating hormone and ultraviolet radiation on the transfer of melanosomes to keratinocytes. AB - The epidermal melanin unit in human skin is composed of melanocytes and keratinocytes. Melanocytes, located in the basal layer of the epidermis, manufacture melanin-loaded organelles called melanosomes. Through their dendritic processes, melanocytes distribute melanosomes to neighboring keratinocytes, where their presence confers to the skin its characteristic color and photoprotective properties. In this study, we used murine melanocytes and keratinocytes alone and in coculture to characterize the processes involved in melanosome transfer. Ultraviolet (UV) radiation induced an accumulation of melanosomes in melanocytes, whereas treatment with a-melanocyte-stimulating hormone (MSH) induced exocytosis of melanosomes accompanied by ruffling of the melanocyte membrane. We found that keratinocytes phagocytose melanosomes and latex beads equally well and that this phagocytic process was increased by exposure of keratinocytes to UV radiation or to MSH. Coculture of melanocytes and keratinocytes resulted in an increase in MSH released to the medium. Gene array analysis of MSH-treated melanocytes showed up regulation of many genes associated with exocytosis. In our studies, we never observed cytophagocytosis of melanosome-filled processes. This result, together with the other findings, suggests that a combination of signals that increase melanosome production and release by melanocytes and that stimulate phagocytosis by keratinocytes are the most relevant mechanisms involved in skin tanning. PMID- 11729102 TI - Angiopoietin-1 negatively regulates expression and activity of tissue factor in endothelial cells. AB - Normally, tissue factor (TF) is not expressed on the surface of endothelial cells, but its expression can be induced by vascular endothelial growth factor (VEGF) and tumor necrosis factor (TNF)-a. However, the signaling pathway(s) affecting this induction is unknown. Using human umbilical vein endothelial cells, we found that inhibitors of guanine-cytosine-rich DNA binding protein and nuclear factor (NF)-kB suppressed VEGF- and TNF-a-induced expression and activity of TF. However, unexpectedly, phosphatidylinositol (PI) 3'-kinase inhibitor enhanced the VEGF- and TNF-a-induced expression and activity of TF. Angiopoietin 1 (Ang1), a strong activator of intracellular PI 3'-kinase/Akt, inhibited the induction of TF by VEGF and TNF-a, whereas Ang1 itself did not produce any significant effect on TF. Selective activation (or inactivation) of PI 3' kinase/Akt by using adenoviral transfer reduced (or enhanced) TNF-a-induced expression of TF mRNA and protein, regardless of Ang1 treatment. From these results, we conclude that Ang1 inhibits the up-regulation of TF expression, possibly through activation of PI 3'-kinase/Akt in endothelial cells. Ang1 may be useful as an inhibitor of VEGF- and TNF-a-induced coagulation, inflammation, and cancer progression. PMID- 11729103 TI - p38 Mitogen-activated protein kinase and phosphatidylinositol 3-kinase activities have opposite effects on human neutrophil apoptosis. AB - Neutrophil apoptosis is essential for resolution of inflammatory reactions. Here, we studied the role of two apoptosis/survival-associated protein kinases in this process. We discovered a previously undetected early and transient inhibition of the activity of p38 mitogen-activated protein kinase (p38 MAPK) during both spontaneous and Fas-induced apoptosis. Pharmacological inhibition of this enzyme augmented the activation of caspases and the apoptotic response, which suggests that the p38 MAPK signals survival in neutrophils. Our finding that caspase-3 activity was initiated during the transient inhibition of p38 MAPK suggests that apoptosis is initiated during this inhibition. Furthermore, such transient inhibition was counteracted by granulocyte-macrophage colony-stimulating factor, which elicits survival. We also found that neither this inhibition of p38 MAPK nor the spontaneous apoptotic response depended on Fas. Instead, the early inhibition of p38 MAPK concurred with a Fas-induced activation of phosphatidylinositol 3-kinase, inhibition of which reduced apoptosis. Thus, the Fas-induced augmentation of spontaneous apoptosis can be explained by its activation of phosphatidylinositol 3-kinase. We conclude that p38 MAPK activity represents a survival signal that is inactivated transiently during both spontaneous and Fas-induced apoptosis, whereas Fas-induced phosphatidylinositol 3 kinase activity is a proapoptotic signal in isolated human neutrophils. PMID- 11729105 TI - WNT1 inducible signaling pathway protein 3, WISP-3, a novel target gene in colorectal carcinomas with microsatellite instability. AB - BACKGROUND & AIMS: Microsatellite instability (MSI) is the phenotype of colorectal carcinomas with defect mismatch repair. Genes with repetitive sequences within their coding regions are targets for mutations in these tumors. We have evaluated 2 novel candidate genes for potential involvement in development of MSI colorectal carcinomas and compared them with alterations in known target genes. METHODS: The MSI status was determined by multiplex polymerase chain reactions (PCRs) of 5-17 markers in a Norwegian series of 275 colorectal carcinomas. All MSI tumors were analyzed for gene mutations using fluorescence PCR followed by capillary electrophoresis. Two novel candidate genes, WNT1-inducible signaling pathway protein 3 (WISP-3) and caspase-1, and 9 known target genes were analyzed. RESULTS: Thirteen percent of the tumors were MSI-high (H) and 12% were MSI-low (L). Thirty-three of 37 MSI-H vs. 1 of 34 MSI-L tumors showed mutations in the target genes (P < 0.001). WISP-3 was mutated in 31% of the MSI-H tumors. The frequencies of frameshift mutations in the known target genes were comparable with other studies. CONCLUSIONS: The relative high frequency of mutation, higher than those seen for other known target genes, the predicted truncation of the protein product, and the homology with WISP-1 and WISP-2, 2 proteins induced downstream of WNT1 signaling, strongly suggest WISP-3 as a novel target in development of MSI-H colorectal carcinomas. PMID- 11729106 TI - Gastric surgery is not a risk for Barrett's esophagus or esophageal adenocarcinoma. AB - BACKGROUND & AIMS: The contribution of duodeno-gastroesophageal reflux to the development of Barrett's esophagus has remained an interesting but controversial topic. The present study assessed the risk for Barrett's esophagus after partial gastrectomy. METHODS: The data of outpatients from a medicine and gastroenterology clinic who underwent upper gastrointestinal endoscopy for any reason were analyzed in a case-control study. A case population of 650 patients with short- segment and 366 patients with long-segment Barrett's esophagus was compared in a multivariate logistic regression to a control population of 3047 subjects without Barrett's esophagus or other types of gastroesophageal reflux disease. RESULTS: In the case population, 25 (4%) patients with short-segment and 15 (4%) patients with long-segment Barrett's esophagus presented with a history of gastric surgery compared with 162 (5%) patients in the control population, yielding an adjusted odds ratio of 0.89 with a 95% confidence interval of 0.54 1.46 for short-segment and an adjusted odds ratio of 0.71 (0.30-1.72) for long segment Barrett's esophagus. Similar results were obtained in separate analyses of 64 patients with Billroth-1 gastrectomy, 105 patients with Billroth-2 gastrectomy, and 33 patients with vagotomy and pyloroplasty for both short- and long-segment Barrett's esophagus. Caucasian ethnicity, the presence of hiatus hernia, and alcohol consumption were all associated with elevated risks for Barrett's esophagus. CONCLUSIONS: Gastric surgery for benign peptic ulcer disease is not a risk factor for either short- or long-segment Barrett's esophagus. This lack of association between gastric surgery and Barrett's esophagus suggests that reflux of bile without acid is not sufficient to damage the esophageal mucosa. PMID- 11729107 TI - Risk of adenocarcinomas of the esophagus and gastric cardia in patients with gastroesophageal reflux diseases and after antireflux surgery. AB - BACKGROUND & AIMS: Gastroesophageal reflux has been proposed as an important risk factor for esophageal and gastric cardia adenocarcinoma, but prospective data are lacking. Furthermore, the effect of antireflux surgery has not yet been studied. We conducted a population-based retrospective cohort study to fill these gaps. METHODS: A cohort of 35,274 male and 31,691 female patients with a discharge diagnosis of gastroesophageal reflux diseases, and another cohort of 6406 male and 4671 female patients who underwent antireflux surgery, were identified in the Swedish Inpatient Register. Follow-up was attained through record linkage with several nationwide registers. Standardized incidence ratio (SIR) was used to estimate relative risk of upper gastrointestinal cancers, using the general Swedish population as reference. RESULTS: After exclusion of the first year follow-up, 37 esophageal and 36 gastric cardia adenocarcinomas were observed among male patients who did not have surgery (SIR, 6.3, 95% confidence interval [CI], 4.5-8.7; SIR, 2.4, 95% CI, 1.7-3.3, respectively). SIR for esophageal adenocarcinoma increased with follow-up time (P = 0.03 for trend). Among male patients who had undergone antireflux surgeries, risks were also elevated (16 esophageal adenocarcinoma, SIR, 14.1, 95% CI, 8.0-22.8; 15 gastric cardia adenocarcinomas, SIR, 5.3, 95% CI, 3.0-8.7) and remained elevated with time after surgery. The cancer risk pattern in women was similar to that for men, but the number of cases were much smaller. CONCLUSIONS: Gastroesophageal reflux is strongly associated with the risk of esophageal adenocarcinoma, and to a lesser extent, with gastric cardia adenocarcinoma. The risk of developing adenocarcinomas of the esophagus and gastric cardia remains increased after antireflux surgery. PMID- 11729108 TI - Extraesophageal associations of gastroesophageal reflux disease in children without neurologic defects. AB - BACKGROUND & AIMS: The potential association between gastroesophageal reflux disease (GERD) and extraesophageal manifestations remains unknown in children without neurological defects. We conducted a large case-control study to examine the association between GERD and several upper and lower respiratory disorders in these children. METHODS: We identified all patients between 2 and 18 years of age with GERD who were seen at Texas Children's Hospital between 1996 and 2000. Patients seen during the same time period without GERD were randomly selected as controls (4:1 ratio). Patients with mental retardation, cerebral palsy, or congenital esophageal anomalies were excluded. We compared the presence of several predefined upper and lower respiratory disorders in cases and controls. RESULTS: We identified 1980 patients with GERD and 7920 controls without GERD. Cases and controls were without neurological deficits or congenital esophageal anomalies. Cases were older than controls (9.2 years +/- 4.6 vs. 8.6 +/- 4.9, P < 0.0001), and were more likely to be female (51.2% vs. 47.2%, P = 0.0028) and white (60.2% vs. 41.2%, P < 0.0001). Compared with controls in univariate analyses, cases with GERD had more sinusitis (4.2% vs. 1.4%, P < 0.0001), laryngitis (0.7% vs. 0.2%), asthma (13.2% vs. 6.8%, P < 0.0001), pneumonia (6.3% vs. 2.3%, P < 0.0001), and bronchiectasis (1.0% vs. 0.1%, P < 0.0001). However, otitis media was less common in cases than controls (2.1% vs. 4.6%, P < 0.0001). After adjusting for differences in age, gender, and ethnicity in the regression analyses, GERD remained a significant risk factor for sinusitis (adjusted odds ratio [OR], 2.3; 95% confidence intervals [CI], 1.7-3.2; P < 0.0001), laryngitis (OR, 2.6; CI, 1.2-5.6; P = 0.0228), asthma (OR, 1.9; CI, 1.6-2.3; P < 0.0001), pneumonia (OR, 2.3; CI, 1.8-2.9; P < 0.0001), and bronchiectasis (OR, 2.3; CI, 1.1-4.6; P = 0.0193). CONCLUSIONS: GERD in children without neurological defects is associated with a several-fold increase in the risk of sinusitis, laryngitis, asthma, pneumonia, and bronchiectasis. Further studies are needed to examine whether a cause-effect relationship exists between GERD and these disorders in children. PMID- 11729109 TI - Extensive methylation of hMLH1 promoter region predominates in proximal colon cancer with microsatellite instability. AB - BACKGROUND & AIMS: Methylation of the hMLH1 promoter region has been suggested to cause microsatellite instability (MSI) in sporadic colorectal carcinoma (CRC). We studied the methylation profile in a wide region of the hMLH1 promoter and compared with the hMLH1 protein expression and MSI status in 88 cases of sporadic CRC. METHODS: Na-bisulfite treatment and polymerase chain reaction single-strand conformation polymorphism analysis was performed using 5 sets of polymerase chain reaction primers spanning the promoter region of the hMLH1 to examine methylation status. Results were compared with immunostaining using anti-hMLH1 monoclonal antibody and MSI status of the tumor samples. RESULTS: Methylation status was classified as full or partial methylation. Full methylation indicates the methylation of all CpG sites in the examined regions. Methylation of the hMLH1 promoter was observed in 88.9% (16 of 18) of CRCs showing high frequency MSI (MSI H), among which 89% (14 of 16) had full methylation with reduced hMLH1 protein expression. All cases showing full methylation were proximal colon tumors with MSI-H. In cases with partial methylation, only the upstream region of the hMLH1 promoter was methylated. Partial methylation was also shown in 33.3% (6 of 18) of the normal mucosa of MSI-H cases. Frequencies of methylation were significantly correlated with female gender (P = 0.0009) and aging (P = 0.007). CONCLUSIONS: Full methylation of the hMLH1 promoter region and subsequent gene inactivation may play a crucial role in the carcinogenesis of MSI-H CRCs in the proximal colon. Methylation upstream of the hMLH1 promoter appears to be an early event in the carcinogenesis of MSI-H tumors. PMID- 11729110 TI - Cystic fibrosis gene mutations and pancreatitis risk: relation to epithelial ion transport and trypsin inhibitor gene mutations. AB - BACKGROUND & AIMS: Nonalcoholic chronic pancreatitis is usually idiopathic and often associated with cystic fibrosis gene (CFTR) mutations. It is unknown whether pancreatitis risk correlates with having 1 or 2 CFTR mutations, abnormal epithelial ion transport, or mutations of other genes. METHODS: We tested 39 patients with idiopathic chronic pancreatitis (mean age at diagnosis, 33 years) for common mutations of CFTR and of genes encoding a trypsin inhibitor (PSTI) and trypsinogen (PRSS1). To exclude hereditary pancreatitis, we initially relied on family history and subsequently tested for PRSS1 mutations. Twenty subjects were tested for rare CFTR mutations (DNA sequencing) and 11 were tested for extrapancreatic CFTR function (clinical and physiologic evaluation). RESULTS: Mutations were identified in 24 of 39 subjects. Nine patients had cystic fibrosis causing mutations, 8 of whom also had mild-variable mutations. Eight others had only mild-variable mutations. Nine subjects had the N34S PSTI mutation and 1 had hereditary pancreatitis (R122H, PRSS1). Pancreatitis risk was increased approximately 40-fold by having 2 CFTR mutations (P < 0.0001), 20-fold by having N34S (P < 0.0001), and 900-fold by having both (P < 0.0001). Subjects with 2 CFTR mutations had abnormal nasal epithelial ion transport and clinical findings suggesting residual CFTR function between that in cystic fibrosis and in carriers. By contrast, subjects with only PSTI mutations had normal CFTR function. CONCLUSIONS: CFTR-related pancreatitis risk correlates with having 2 CFTR mutations and reduced extrapancreatic CFTR function. The N34S PSTI mutation increased risk separately. Testing for pancreatitis-associated CFTR and PSTI genotypes may be useful in nonalcoholic pancreatitis. PMID- 11729111 TI - Epithelial barrier defects in ulcerative colitis: characterization and quantification by electrophysiological imaging. AB - BACKGROUND & AIMS: In ulcerative colitis (UC), the epithelial barrier is impaired by erosion/ulcer-type lesions and epithelial apoptosis causing local leaks, and generalized tight junction alterations increasing the basal permeability. We quantified the contribution of these mechanisms to the increased colonic ion permeability. METHODS: Sigmoid colon was stripped, and the spatial distribution of current clamped across the viable epithelium was recorded by a microelectrode probe, using the conductance scanning method. Local leaks (circumscribed conductive peaks) were marked, and structural changes were studied in H&E-stained series sections. RESULTS: Overall conductivity increased from 8.4 +/- 0.7 mS/cm(2) (mean +/- SEM) in controls to 11.7 +/- 0.6 in specimens with mild inflammation (i.e., with intact epithelium) and 34.4 +/- 6.2 mS/cm(2) in moderate to-severe inflammation (i.e., with visible epithelial lesions). Only in part this was caused by a generalized increase in basal conductivity (12.2 +/- 1.5 mS/cm(2) in moderate-to-severe UC vs. 8.3 +/- 0.7 in controls). More importantly, the spatial distribution of conductivity, which was even in controls, showed dramatic leaks in UC. Leaks found in mild inflammation without epithelial lesion turned out to be foci of epithelial apoptosis. In moderate-to-severe inflammation, leaks correlated with epithelial erosion/ulcer-type lesions or crypt abscesses. CONCLUSIONS: In early UC, but not in controls, seemingly intact epithelium comprises leaks at apoptotic foci. With more intensive inflammation, erosion/ulcer-type lesions are highly conductive, even if covered with fibrin. Local leaks contribute 19% to the overall epithelial conductivity in mild and 65% in moderate-to-severe inflammation. PMID- 11729112 TI - Celiac disease-like abnormalities in a subgroup of patients with irritable bowel syndrome. AB - BACKGROUND & AIMS: Abdominal symptoms in the absence of mucosal abnormalities are features of both the irritable bowel syndrome (IBS) and latent/potential celiac disease (cd). To identify a possible subgroup of IBS patients with latent/potential cd, surrogate markers of cd were investigated in IBS patients. METHODS: IBS patients suffering from diarrhea (n = 102), and patients with active cd (n = 10), treated cd (n = 26), and latent cd (n = 5) were included in the study. We measured serum immunoglobulin (Ig) A against gliadin and tissue transglutaminase, and IgA and IgM against gliadin, tissue-transglutaminase (intestinal cd-associated antibodies), and the dietary proteins beta lactoglobulin and ovalbumin in duodenal aspirate by enzyme-linked immunosorbent assay. Intraepithelial lymphocytes (IELs) were counted in histology sections, and the expression of HLA-DQ2 (A1*0501/B1*0201) was investigated by polymerase chain reaction. In 26 IBS patients, the effect of 6 months of gluten withdrawal was examined. RESULTS: Most cd patients expressed HLA-DQ2 and had increased intestinal cd-associated antibodies, whereas cd-associated serum IgA and IEL counts were increased in active cd in contrast to treated or latent cd. In IBS patients, 35% were HLA-DQ2-positive, 23% had increased IEL counts, and 0% and 30% had increased cd-associated antibodies in serum and duodenal aspirate, respectively. Furthermore, stool frequency and intestinal IgA decreased significantly under a gluten-free diet in the subgroups of HLA-DQ2-positive and intestinal antibody-positive IBS patients when compared with IBS patients without these markers. CONCLUSIONS: HLA-DQ2 expression and increased intestinal cd associated antibodies are markers that can identify latent/potential cd in a subgroup of IBS patients who consequently appear to profit from a gluten-free diet. PMID- 11729113 TI - Up-regulation of cyclooxygenase 2 gene expression correlates with tumor angiogenesis in human colorectal cancer. AB - BACKGROUND & AIMS: Recent studies have shown that cyclooxygenase (COX)-2 and its products, prostaglandins (PGs), may be involved in colorectal carcinogenesis. The aim of this study was to determine whether COX-2 expression and PGE(2) production correlate with microvessel density, vascular endothelial growth factor (VEGF) expression, and tumor metastasis in human colorectal cancer. METHODS: Tumor samples and adjacent normal mucosa were obtained from 31 surgical specimens. Immunohistochemical expression of COX-2, VEGF, and CD31 was analyzed on paraffin embedded tissue sections. COX-2 and COX-1 proteins were determined by Western blot analysis. COX-2 and VEGF messenger RNA expressions were evaluated using Northern blot analysis. PGE(2) production was determined by specific radioimmunoassay. RESULTS: The immunohistochemical expressions of both COX-2 and VEGF were significantly correlated with microvessel density (P = 0.02 and P = 0.002, respectively). A significant correlation was found between COX-2 and VEGF expression (P = 0.004). Western analysis confirmed the up-regulation of COX-2 protein expression. COX-2 and VEGF genes were overexpressed in tumor specimens as compared with normal mucosa. PGE(2) levels were significantly higher in metastatic tumors than in nonmetastatic ones (P = 0.03). CONCLUSIONS: COX-2 is related to tumor angiogenesis in colorectal cancer. It is likely that VEGF is one of the most important mediators of the COX-2 angiogenic pathway. PMID- 11729114 TI - Incidence of gastric cancer and breast cancer in CDH1 (E-cadherin) mutation carriers from hereditary diffuse gastric cancer families. AB - BACKGROUND & AIMS: Germline mutations in CDH1 are known to cause hereditary diffuse gastric cancer (HDGC). Breast and colorectal cancer have also been reported in CDH1-associated HDGC. The purpose of this study was to estimate the cumulative risk of gastric and breast cancer in CDH1 mutation carriers. METHODS: Family data were collected by member groups of the International Gastric Cancer Linkage Consortium. Eligible families had at least 3 cases of diffuse gastric cancer, and at least 1 affected member had tested positive for a mutation in CDH1. Eleven families met these criteria. We used the pedigree information to estimate penetrance using the MENDEL program. The conditional likelihood of the pedigree was maximized given the phenotype of the pedigree and genotype of the index case at ascertainment. We parameterized the model in terms of log relative risks for mutation carriers compared with risks in the general population of the United Kingdom. Noncarriers of the gene were assumed to develop the disease at population incidence rates. RESULTS: The estimated cumulative risk of gastric cancer by age 80 years was 67% for men (95% confidence interval [95% CI], 39-99) and 83% for women (95% CI, 58-99). For women, the cumulative risk of breast cancer was 39% (95% CI, 12-84). The combined risk of gastric cancer and breast cancer in women was 90% by age 80 years. CONCLUSIONS: These penetrance estimates should be useful for genetic counseling in multiple-case families. However, they may not apply to individuals with a minimal family history, in whom the risks may be lower. PMID- 11729115 TI - Prostanoid production via COX-2 as a causative mechanism of rodent postoperative ileus. AB - BACKGROUND & AIMS: This study demonstrates a significant role for cyclooxygenase (COX)-2 and prostanoid production as mechanisms for surgically induced postoperative ileus. METHODS: Rats, COX-2+/+, and COX-2-/- mice underwent simple intestinal manipulation. Reverse-transcription polymerase chain reaction and immunohistochemistry were used to detect and localize COX-2 expression. Prostaglandin levels were measured from serum, peritoneal lavage fluid, and muscularis culture media. Jejunal circular muscle contractions were measured in an organ bath, and gastrointestinal transit was measured in vivo. RESULTS: The data show that intestinal manipulation induces COX-2 messenger RNA and protein within resident muscularis macrophages, a discrete subpopulation of myenteric neurons and recruited monocytes. The manipulation-induced increase in COX-2 expression resulted in significantly elevated prostaglandin levels within the circulation and peritoneal cavity. The source of these prostanoids could be directly attributed to their release from the inflamed muscularis externa. As a consequence of the molecular up-regulation of COX-2, we observed a decrease in in vitro jejunal circular muscle contractility and gastrointestinal transit, both of which could be alleviated pharmacologically with selective COX-2 inhibition. These studies were corroborated with the use of COX-2-/- mice. CONCLUSIONS: Prostaglandins, through the induction of COX-2, are major participants in rodent postoperative ileus induced by intestinal manipulation. PMID- 11729116 TI - Blockade of endogenous IL-18 ameliorates TNBS-induced colitis by decreasing local TNF-alpha production in mice. AB - BACKGROUND & AIMS: Interleukin (IL) 18 has proinflammatory effects. IL-18 plays a pivotal role in Th1 responses, but its proinflammatory activities extend beyond Th1 cells, including macrophages and production of tumor necrosis factor (TNF) alpha and IL-1beta. IL-18 is up-regulated in colonic specimens of patients with Crohn's disease. The goal of this study was to evaluate the role of IL-18. METHODS: Activity of IL-18 was neutralized using recombinant human IL-18 binding protein isoform a (rhIL-18BPa) in trinitrobenzene sulfonic acid (TNBS)-induced colitis. RESULTS: Mice treated daily with rhIL-18BPa (8 mg/kg) had significant reductions in clinical score, body weight loss, and colon weight increase compared with saline-treated mice. Histologic analysis showed that rhIL-18BPa treated mice developed only mild colitis without signs of ulceration, with a mean total score of 9.8 +/- 1.3 points compared with 15.9 +/- 1.1 points observed in saline-treated mice with colitis. Analysis of cytokine levels in colon homogenates showed a significant decrease in TNF-alpha, IL-6, and IL-1beta after rhIL-18BPa treatment but no effect on interferon gamma. The therapeutic potential of rhIL-18BPa treatment was confirmed in TNBS mice that were treated only on days 8 and 9 after the start of the experiment. In these mice, significant reductions in total colitis score and colon weight were also observed. CONCLUSIONS: These findings show that inhibition of rhIL-18BPa bioactivity, via rhIL-18BPa, may be beneficial for the treatment of IBD. PMID- 11729117 TI - Human pancreatic acinar cells lack functional responses to cholecystokinin and gastrin. AB - BACKGROUND & AIMS: Pancreatic acinar cells from various species express cholecystokinin (CCK) A, CCK-B, or a combination of these CCK receptor subtypes. The presence and functional roles of CCK receptors on human acinar cells remain unclear. METHODS: Acini isolated from human pancreas were treated with CCK receptor agonists, CCK-8 and gastrin, and an agonist for m3 muscarinic acetylcholine receptors (m3 AchR), carbachol. Functional parameters measured included intracellular [Ca(2+)], amylase secretion, and ERK phosphorylation. Binding studies were performed using (125)I-CCK-8. Expression of messenger RNAs (mRNAs) was determined using real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) and localized by in situ hybridization. RESULTS: Human acini did not respond to CCK agonists. In contrast, they responded to carbachol with robust increases in each of the functional parameters. Moreover, the cells responded to CCK agonists after adenoviral-mediated gene transfer of CCK-A or CCK-B receptors. A low level of specific and a high level of nonspecific binding of (125)I-CCK-8 were observed. Quantitative RT-PCR indicated that the message levels for CCK-A receptors were approximately 30-fold lower than those of CCK-B receptors, which were approximately 10-fold lower than those of m3 Ach receptors. In situ hybridization indicated the presence of m3 Ach receptor and insulin mRNA but not CCK-A or CCK-B receptor mRNAs in adult human pancreas. CONCLUSIONS: These data indicate that human pancreatic acinar cells do not respond to CCK receptor agonists in terms of expected functional parameters and show that this is due to an insufficient level of receptor expression. PMID- 11729118 TI - Duodenal reflux induces cyclooxygenase-2 in the esophageal mucosa of rats: evidence for involvement of bile acids. AB - BACKGROUND & AIMS: Reflux of duodenal contents including bile acids is believed to contribute to esophageal injury and Barrett's esophagus. Cyclooxygenase (COX) 2, an inducible form of COX, has been implicated in both inflammation and carcinogenesis. In this study, we investigated the effects of bile acids and duodenal reflux on COX-2 expression in cultured esophageal cells and tissue, respectively. METHODS: Immunoblotting and Northern blotting were used to assess the effects of bile acids on COX-2 expression in esophageal cell lines. Immunoblotting and immunohistochemistry were performed to evaluate the effects of duodenal reflux on COX-2 expression and cell proliferation in esophageal tissue. RESULTS: Unconjugated bile acids were about fivefold more potent inducers of COX 2 messenger RNA, COX-2 protein, and prostaglandin synthesis than conjugated bile acids. Acidifying the culture medium sensitized esophageal cells to bile acid mediated induction of COX-2. The induction of COX-2 by bile acids was mediated by phosphatidylinositol-3 kinase and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinases. In experimental animals, duodenoesophageal reflux led to esophagitis, marked thickening of the esophageal mucosa, and enhanced expression of COX-2. Increased immunoreactivity for Ki-67 and cyclin D1 indicated that enhanced cell proliferation contributed to mucosal thickening. CONCLUSIONS: Reflux of duodenal contents into the esophagus led to increased COX 2 expression and mucosal thickening. Bile acids are likely to contribute to these effects. PMID- 11729119 TI - D-glucose releases 5-hydroxytryptamine from human BON cells as a model of enterochromaffin cells. AB - BACKGROUND & AIMS: 5-Hydroxytryptamine (5-HT) is released from enterochromaffin cells and activates neural reflex programs regulating motility and secretion. Although sugars are reported to release 5-HT in vivo, it is unclear whether they act directly on enterochromaffin cells or indirectly through an intermediary messenger. The aim was to determine if D-glucose is a stimulus for 5-HT release. METHODS: Human BON cells, derived from enterochromaffin cells, were treated with D-glucose, galactose, and the nonmetabolizable methyl alpha-D-glucopyranoside, or with fructose. RESULTS: Reverse-transcription polymerase chain reaction together with Western blot analysis revealed an SGLT-like protein. D-glucose caused a concentration-dependent increase in 5-HT release, which was mimicked by methyl alpha-D-glucopyranoside and galactose but not fructose. D-glucose-stimulated 5-HT release was significantly reduced by phloridzin. Concentrations of mannitol below 75 mmol/L were ineffective in releasing 5-HT. Brefeldin A abolished forskolin stimulated 5-HT release without affecting basal or constitutive release. CONCLUSIONS: The results show that high concentrations of metabolizable and nonmetabolizable hexoses activate signal transduction pathways, leading to release of 5-HT. These findings imply a role for enterochromaffin cells as "glucose sensors" during ingestion of a meal. PMID- 11729120 TI - The role of mutant Apc in the development of dysplasia and cancer in the mouse model of dextran sulfate sodium-induced colitis. AB - BACKGROUND & AIMS: Differences in genetic background may play a role in the development of ulcerative colitis (UC)-related neoplasia. Loss of heterozygosity (LOH) of APC has been reported in human UC-associated neoplasia. To investigate the role of genetic differences in UC-associated neoplasia, we compared differences in dextran sodium sulfate (DSS) colitis-associated neoplasia between wild-type C57BL/6J mice (WT-DSS) and C57BL/6J mice with a germline mutation in Apc (Min-DSS). METHODS: DSS colitis was induced in female wild-type and Min mice. Age- and sex-matched non-DSS-treated Mins were also studied. Animals were sacrificed after 1 and 2 cycles of DSS. The cecums and large intestines were studied for numbers of dysplasias/cancers. Dysplasias were studied for LOH of Apc. RESULTS: No WT-DSS, 100% of Min-DSS, and 50% of non-DSS-treated Mins had dysplasia. The mean numbers of lesions per mouse were 0 (WT-DSS), 15.6 and 29.3 (1 and 2 cycles Min-DSS, respectively), 1.2 and 1.9 (age-matched control Min, 1 and 2 cycle equivalents, respectively; P < 0.0002, Min-DSS vs. WT-DSS and non-DSS treated Min; P = 0.03, Min-DSS 2 cycle vs. Min-DSS 1 cycle). Cancers were seen in 0%, 22%, and 40% of non-DSS Min, Min-DSS-1 cycle, and Min-DSS-2 cycle animals, respectively. LOH of Apc was observed in 90.6% of dysplasias and 6% of nondysplastic mucosa. CONCLUSIONS: A germline mutation in Apc contributes significantly to the development of colitis-associated neoplasia. Colitis markedly accelerates the development of dysplasia and cancer in the Min mouse. Dysplasia in Min-DSS occurs through LOH of Apc. PMID- 11729121 TI - Antral mucosa expresses functional leptin receptors coupled to STAT-3 signaling, which is involved in the control of gastric secretions in the rat. AB - BACKGROUND & AIMS: Leptin is a circulating hormone that communicates the peripheral nutritional status to the hypothalamus, which controls food intake, energy expenditure, and body weight. This study characterizes leptin receptors and leptin-sensitive STAT proteins in the antrum and investigates the effects of leptin on gastric secretions. METHODS: The effects of leptin on gastrin messenger RNA (mRNA), plasma gastrin, gastric acid in vivo in the rat, and on somatostatin and gastrin secretions by isolated antral cells were determined in vitro. Leptin receptors were investigated in isolated rat antral cells by reverse transcription polymerase chain reaction and binding of [(125)I]-leptin studies. The effects of in vivo and in vitro leptin on transduction signal STAT proteins were investigated by immunoblotting antral extracts. RESULTS: Peripheral injection of leptin inhibited in a dose-dependent manner, basal gastric secretion, gastrinemia, and mucosal gastrin mRNA in vivo. mRNAs encoding the long (Ob-Rb) and short (Ob-Ra) receptor forms were detected in rat antral mucosa, as were STAT 1, -3, and -5b immunoreactive proteins. Isolated antral cells specifically bound [(125)I]-leptin, and addition of leptin to these cells inhibited the release of somatostatin and increased the release of gastrin. These effects were associated with an increase in nuclear STAT-3 proteins in vitro and in vivo. CONCLUSIONS: This study provides the first molecular evidence for the coexpression of leptin receptors and STAT-3 in antral mucosa. It provides further evidence for the involvement of leptin in the control of gastric secretions. PMID- 11729122 TI - Antibody blockade of ICAM-1 and VCAM-1 ameliorates inflammation in the SAMP-1/Yit adoptive transfer model of Crohn's disease in mice. AB - BACKGROUND & AIMS: Integrins (alpha(4) and beta(2)) and their endothelial ligands vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) play key roles in leukocyte recruitment to areas of inflammation. ICAM-1 and VCAM-1 are expressed in inflamed intestinal tissues. This study investigates a possible causative role of adhesion molecules ICAM-1, VCAM-1, and alpha(4) integrins in mediating the inflammatory response in a murine model of Crohn's disease (CD). METHODS: CD4+ mesenteric lymph node cells from SAMP-1/Yit donor mice were adoptively transferred into major histocompatibility complex-matched severe combined immunodeficiency disease mice. Six weeks later, these mice were left untreated or treated for 3 days with monoclonal antibodies (mAbs) to ICAM-1, VCAM-1, or both, and alpha(4), or both ICAM-1 and alpha(4), dexamethasone, or nonblocking isotype control antibodies. On day 4 after treatment, tissues were investigated for expression of ICAM-1, VCAM-1, and for severity of inflammation using a semiquantitative inflammatory score. Dexamethasone treatment resolved all measures of intestinal inflammation. RESULTS: Blocking either ICAM-1, VCAM-1, or alpha(4) integrins had no significant beneficial effect. However, blocking ICAM-1 and alpha(4), or blocking ICAM-1 and VCAM-1, showed a 70% resolution of the active inflammation, but not chronic inflammation. CONCLUSIONS: These findings suggest that blocking ICAM-1 and VCAM-1 may have therapeutic benefit for the acute inflammatory component of Crohn's disease. PMID- 11729123 TI - Phosphorylation of the serine 60 residue within the Cdx2 activation domain mediates its transactivation capacity. AB - BACKGROUND & AIMS: Cdx2 is critical in intestinal proliferation and differentiation. Modulation of Cdx2 function in response to cellular signaling is to be elucidated. We hypothesize that phosphorylation of the Cdx2 activation domain can modulate its function. METHODS: The Cdx2 activation domain was delineated in transient transfections using different portions of Cdx2 fused to the Gal4-DNA binding domain. In vivo phosphorylation was studied by metabolic labeling with (32)P-orthophosphate. To study a potential phosphorylation site, polyclonal antibodies were generated: CNL was raised against amino acids 54-66 of Cdx2 and P-Cdx2-S60 against the same epitope in which serine 60 was phosphorylated. RESULTS: A critical region for transactivation resides within amino acids 60-70. Substitution of serine 60 with alanine reduces incorporation of (32)P-orthophosphate substantially. S60-phosphorylation decreases Cdx2 transactivation. Phosphorylation of serine 60 can be inhibited with the mitogen activated protein kinase inhibitors PD98059 or UO126. P-Cdx2-S60 recognizes phosphorylated serine 60 mainly in proliferative compartment of the intestinal epithelial layer. In contrast, CNL recognizes Cdx2 predominantly in the differentiated compartment. CONCLUSIONS: The Cdx2 activation domain is phosphorylated at serine 60 via the mitogen-activated protein kinase pathway. S60 phosphorylated and S60-nonphosphorylated Cdx2 have different transcriptional activity, as well as different spatial expression patterns in the intestinal epithelium. PMID- 11729124 TI - The superoxide dismutase mimetic MnTBAP prevents Fas-induced acute liver failure in the mouse. AB - BACKGROUND & AIMS: Acute liver failure (ALF) of viral origin results from massive hepatocyte apoptosis induced by the interaction between Fas expressed on hepatocytes and Fas ligand on activated T lymphocytes. Because Fas-induced apoptosis of hepatocytes involves mitochondrial damages and potential reactive oxygen species (ROS) overproduction, we investigated whether manganese III tetrakis (5,10,15,20 benzoic acid) (MnTBAP), a nonpeptidyl mimic of superoxide dismutase (SOD), can inhibit Fas-induced ALF. METHODS: An agonist anti-Fas monoclonal antibody was used to induce hepatocyte apoptosis in vitro and ALF in vivo. RESULTS: Preventive and curative treatments by MnTBAP significantly increased survival rates and significantly reduced aspartate aminotransferase levels and parenchymal lesions. ROS generation was suggested by those beneficial effects and significant increases in SOD and Gpx activities after anti-Fas injection. Flow cytometry of isolated hepatocytes incubated with anti-Fas monoclonal antibody showed that ROS production was associated with the collapse of transmembrane potential and loss of cardiolipin content. After injection of anti-Fas monoclonal antibody, mitochondrial Bcl-2 was decreased, cytochrome c released, and caspase-3 activated. Mitochondrial alterations and their consequences were abrogated by MnTBAP. CONCLUSIONS: ROS are key executioners in Fas-induced hepatocyte apoptosis. This finding explains why a nonpeptidyl mimic of SOD can cure ALF in a model of viral hepatitis, pointing out the potential interest of this molecule in humans. PMID- 11729125 TI - Kupffer cell-mediated recruitment of rat dendritic cells to the liver: roles of N acetylgalactosamine-specific sugar receptors. AB - BACKGROUND & AIMS: Tissue recruitment of dendritic cells (DCs) is essential for antigen presentation. This study aimed to examine cellular and molecular mechanisms for DC recruitment to the liver. METHODS: Purified rat DCs were injected into circulation and their traffics were analyzed in normal and Kupffer cell-depleted rats by intravital confocal microscopy and immunohistology. Affinities of DCs to sinusoidal cells were examined by a cell-binding assay. DC precursor recruitment was induced by particulate injection. RESULTS: Both DC precursors and DCs at the antigen-transporting stage could be recruited to the liver, and their majority initially showed a selective binding to Kupffer cells. In the Kupffer cell-depleted rats, DCs could neither be recruited to the liver nor adhere to sinusoidal walls. Pretreatment with varied monosaccharides showed that sugar residues consisting of N-acetylgalactosamine were necessary for this binding. The binding was calcium-dependent, implying the C-type lectin involvement. Furthermore, DCs could endocytose N-acetylgalactosamine polymers in a receptor-specific manner. CONCLUSIONS: The DC-Kupffer cell binding through N acetylgalactosamine-specific C-type lectin-like receptors is crucial for DC recruitment to the liver. Rat DCs at least partly possess receptors for endocytosis of galactosylated antigens. These DC receptors as well as Kupffer cell lectins are presumably responsible for this binding. PMID- 11729126 TI - Adaptive regulation of bile salt transporters in kidney and liver in obstructive cholestasis in the rat. AB - BACKGROUND & AIMS: Cholestasis results in adaptive regulation of bile salt transport proteins in hepatocytes that may limit liver injury. However, it is not known if changes also occur in the expression of bile salt transporters that reside in extrahepatic tissues, particularly the kidney, which might facilitate bile salt excretion during obstructive cholestasis. METHODS: RNA and protein were isolated from liver and kidney 14 days after common bile duct ligation in rats and assessed by RNA protection assays, Western analysis, and tissue immunofluorescence. Sodium-dependent bile salt transport was also measured in brush border membrane vesicles from the kidney. RESULTS: After common bile duct ligation, serum bile salts initially rose and then declined to lower levels after 3 days. In contrast, urinary bile salt excretion rose progressively over the 2 week period. By that time, the ileal sodium-dependent bile salt transporter messenger RNA and protein expression in total liver had increased to 300% and 200% of controls, respectively, while falling to 46% and 37% of controls, respectively, in the kidney. Sodium-dependent uptake of (3)H-taurocholate in renal brush border membrane vesicles was decreased. In contrast, the multidrug resistance-associated protein 2 expression in the kidney was increased 2-fold, even 1 day after ligation. Immunofluorescent studies confirmed the changes in the expression of these transporters in liver and kidney. CONCLUSIONS: These studies show that the molecular expression of bile salt transporters in the kidney and cholangiocytes undergo adaptive regulation after common bile duct obstruction in the rat. These responses may facilitate extrahepatic pathways for bile salt excretion during cholestasis. PMID- 11729127 TI - Treatment of vitamin D deficiency due to Crohn's disease with tanning bed ultraviolet B radiation. AB - In Crohn's disease, severe skeletal demineralization, secondary hyperparathyroidism, and muscle weakness can occur. This may be caused by impaired vitamin D absorption, resulting from extensive intestinal disease and resection of duodenum and jejunum, where vitamin D is absorbed. We report a 57 year-old woman with a long history of Crohn's disease and short-bowel syndrome who had only 2 feet of small intestine remaining after 3 bowel resections. She was taking a daily multivitamin containing 400 IU of vitamin D(3) and was dependent on total parenteral nutrition that contained 200 IU of vitamin D and calcium (18 mEq in a 1-L bag infused over 8 hours daily) for a period of 36 months. Despite the above replacement, she complained of bone pain and muscle weakness, and she continued to be vitamin D-deficient with a 25(OH)D level <20 ng/mL. She was then exposed to ultraviolet B (UVB) radiation in a tanning bed wearing a 1-piece bathing suit for 10 minutes, 3 times a week for 6 months at the General Clinical Research Center, Boston University Medical Center. She tolerated the irradiation well without evidence of erythema. After 4 weeks, her serum 25(OH)D level increased by 357% from 7 to 32 ng/mL, parathyroid hormone level decreased by 52% from 92 to 44 pg/mL, and the serum calcium level increased from 7.8 to 8.5 mg/dL. After 6 months of UVB treatment, her serum 25(OH)D level was maintained in the normal range and was free of muscle weakness, and bone and muscle pain. PMID- 11729128 TI - Molecular aspects of iron absorption and HFE expression. AB - Hereditary hemochromatosis, a disease of iron overload, occurs in about 1 in 200 400 Caucasians. The gene mutated in this disorder is termed HFE. The product of this gene, HFE protein, is homologous to major histocompatibility complex class I proteins, but HFE does not present peptides to T cells. Based on recent structural, biochemical, and cell biological studies, transferrin receptor (TfR) is a ligand for HFE. This association directly links HFE protein to the TfR mediated regulation of iron homeostasis. Although evidence is accumulating that binding of HFE to TfR is critical for the effects of HFE, the final pieces in the HFE puzzle have not been established. This review focuses on recent advances in HFE research and presents a hypothetical model of HFE function. PMID- 11729129 TI - Multistep progression of colorectal cancer in the setting of microsatellite instability: new details and novel insights. PMID- 11729130 TI - Acid and bile reflux in Barrett's esophagus: a tale of two evils. PMID- 11729131 TI - Antireflux surgery and adenocarcinoma of the esophagus: let the truth be told. PMID- 11729132 TI - Gene mutations and idiopathic chronic pancreatitis: clinical implications and testing. PMID- 11729133 TI - Irritable bowel syndrome: how far do you go in the workup? PMID- 11729134 TI - Does surveillance endoscopy improve life expectancy in those with Barrett's esophagus? PMID- 11729135 TI - The importance of autoimmune pancreatitis. PMID- 11729136 TI - Human M cells delivered in a box. PMID- 11729137 TI - Effectiveness of colonoscopy in preventing colorectal cancer. PMID- 11729140 TI - Shannon's brief foray into genetics. PMID- 11729141 TI - A filamentous growth response mediated by the yeast mating pathway. AB - Haploid cells of the budding yeast Saccharomyces cerevisiae respond to mating pheromones by arresting their cell-division cycle in G1 and differentiating into a cell type capable of locating and fusing with mating partners. Yeast cells undergo chemotactic cell surface growth when pheromones are present above a threshold level for morphogenesis; however, the morphogenetic responses of cells to levels of pheromone below this threshold have not been systematically explored. Here we show that MATa haploid cells exposed to low levels of the alpha factor mating pheromone undergo a novel cellular response: cells modulate their division patterns and cell shape, forming colonies composed of filamentous chains of cells. Time-lapse analysis of filament formation shows that its dynamics are distinct from that of pseudohyphal growth; during pheromone-induced filament formation, daughter cells are delayed relative to mother cells with respect to the timing of bud emergence. Filament formation requires the RSR1(BUD1), BUD8, SLK1/BCK1, and SPA2 genes and many elements of the STE11/STE7 MAP kinase pathway; this response is also independent of FAR1, a gene involved in orienting cell polarization during the mating response. We suggest that mating yeast cells undergo a complex response to low levels of pheromone that may enhance the ability of cells to search for mating partners through the modification of cell shape and alteration of cell-division patterns. PMID- 11729142 TI - Dual mutations reveal interactions between components of oxidative phosphorylation in Kluyveromyces lactis. AB - Loss of mtDNA or mitochondrial protein synthesis cannot be tolerated by wild-type Kluyveromyces lactis. The mitochondrial function responsible for rho(0)-lethality has been identified by disruption of nuclear genes encoding electron transport and F(0)-ATP synthase components of oxidative phosphorylation. Sporulation of diploid strains heterozygous for disruptions in genes for the two components of oxidative phosphorylation results in the formation of nonviable spores inferred to contain both disruptions. Lethality of spores is thought to result from absence of a transmembrane potential, Delta Psi, across the mitochondrial inner membrane due to lack of proton pumping by the electron transport chain or reversal of F(1)F(0)-ATP synthase. Synergistic lethality, caused by disruption of nuclear genes, or rho(0)-lethality can be suppressed by the atp2.1 mutation in the beta-subunit of F(1)-ATPase. Suppression is viewed as occurring by an increased hydrolysis of ATP by mutant F(1), allowing sufficient electrogenic exchange by the translocase of ADP in the matrix for ATP in the cytosol to maintain Delta Psi. In addition, lethality of haploid strains with a disruption of AAC encoding the ADP/ATP translocase can be suppressed by atp2.1. In this case suppression is considered to occur by mutant F(1) acting in the forward direction to partially uncouple ATP production, thereby stimulating respiration and relieving detrimental hyperpolarization of the inner membrane. Participation of the ADP/ATP translocase in suppression of rho(0)-lethality is supported by the observation that disruption of AAC abolishes suppressor activity of atp2.1. PMID- 11729143 TI - The Kar3-interacting protein Cik1p plays a critical role in passage through meiosis I in Saccharomyces cerevisiae. AB - Meiosis I in Saccharomyces cerevisiae is dependent upon the motor protein Kar3. Absence of Kar3p in meiosis results in an arrest in prophase I. Cik1p and Vik1p are kinesin-associated proteins known to modulate the function of Kar3p in the microtubule-dependent processes of karyogamy and mitosis. Experiments were performed to determine whether Cik1p and Vik1p are also important for the function of Kar3p during meiosis. The meiotic phenotypes of a cik1 mutant were found to be similar to those of kar3 mutants. Cells without Cik1p exhibit a meiotic defect in homologous recombination and synaptonemal complex formation. Most cik1 mutant cells, like kar3 mutants, arrest in meiotic prophase; however, in cik1 mutants this arrest is less severe. These data are consistent with the model that Cik1p is necessary for some, but not all, of the roles of Kar3p in meiosis I. vik1 mutants sporulate at wild-type levels, but have reduced spore viability. This loss in viability is partially attributable to vegetative chromosome loss in vik1 diploids. Cellular localization experiments reveal that Kar3p, Cik1p, and Vik1p are present throughout meiosis and are consistent with Cik1p and Vik1p having different meiotic roles. PMID- 11729144 TI - Dissection of the functions of the Saccharomyces cerevisiae RAD6 postreplicative repair group in mutagenesis and UV sensitivity. AB - The RAD6 postreplicative repair group participates in various processes of DNA metabolism. To elucidate the contribution of RAD6 to starvation-associated mutagenesis, which occurs in nongrowing cells cultivated under selective conditions, we analyzed the phenotype of strains expressing various alleles of the RAD6 gene and single and multiple mutants of the RAD6, RAD5, RAD18, REV3, and MMS2 genes from the RAD6 repair group. Our results show that the RAD6 repair pathway is also active in starving cells and its contribution to starvation associated mutagenesis is similar to that of spontaneous mutagenesis. Epistatic analysis based on both spontaneous and starvation-associated mutagenesis and UV sensitivity showed that the RAD6 repair group consists of distinct repair pathways of different relative importance requiring, besides the presence of Rad6, also either Rad18 or Rad5 or both. We postulate the existence of four pathways: (1) nonmutagenic Rad5/Rad6/Rad18, (2) mutagenic Rad5/Rad6 /Rev3, (3) mutagenic Rad6/Rad18/Rev3, and (4) Rad6/Rad18/Rad30. Furthermore, we show that the high mutation rate observed in rad6 mutants is caused by a mutator different from Rev3. From our data and data previously published, we suggest a role for Rad6 in DNA repair and mutagenesis and propose a model for the RAD6 postreplicative repair group. PMID- 11729145 TI - The Spo12 protein of Saccharomyces cerevisiae: a regulator of mitotic exit whose cell cycle-dependent degradation is mediated by the anaphase-promoting complex. AB - The Spo12 protein plays a regulatory role in two of the most fundamental processes of biology, mitosis and meiosis, and yet its biochemical function remains elusive. In this study we concentrate on the genetic and biochemical analysis of its mitotic function. Since high-copy SPO12 is able to suppress a wide variety of mitotic exit mutants, all of which arrest with high Clb-Cdc28 activity, we speculated whether SPO12 is able to facilitate exit from mitosis when overexpressed by antagonizing mitotic kinase activity. We show, however, that Spo12 is not a potent regulator of Clb-Cdc28 activity and can function independently of either the cyclin-dependent kinase inhibitor (CDKi), Sic1, or the anaphase-promoting complex (APC) regulator, Hct1. Spo12 protein level is regulated by the APC and the protein is degraded in G1 by an Hct1-dependent mechanism. We also demonstrate that in addition to localizing to the nucleus Spo12 is a nucleolar protein. We propose a model where overexpression of Spo12 may lead to the delocalization of a small amount of Cdc14 from the nucleolus, resulting in a sufficient lowering of mitotic kinase levels to facilitate mitotic exit. Finally, site-directed mutagenesis of highly conserved residues in the Spo12 protein sequence abolishes both its mitotic suppressor activity as well as its meiotic function. This result is the first indication that Spo12 may carry out the same biochemical function in mitosis as it does in meiosis. PMID- 11729146 TI - Isolation of X and Y chromosome-specific DNA markers from a liverwort, Marchantia polymorpha, by representational difference analysis. AB - The liverwort Marchantia polymorpha has X and Y chromosomes in the respective female and male haploids. Here we report the successful exploitation of representational difference analyses to isolate DNA markers for the sex chromosomes. Two female-specific and six male-specific DNA fragments were genetically confirmed to originate from the X and Y chromosomes, respectively. PMID- 11729147 TI - K(+)-channel transgenes reduce K(+) currents in Paramecium, probably by a post translational mechanism. AB - PAK11 is 1 of more than 15 members in a gene family that encodes K(+)-channel pore-forming subunits in Paramecium tetraurelia. Microinjection of PAK11 DNA into macronuclei of wild-type cells results in clonal transformants that exhibit hyperexcitable swimming behaviors reminiscent of certain loss-of-K(+)-current mutants. PAK2, a distant homolog of PAK11, does not have the same effect. But PAK1, a close homolog of PAK11, induces the same hyperexcitability. Cutting the PAK11 open reading frame (ORF) with restriction enzymes before injection removes this effect entirely. Microinjection of PAK11 ORF flanked by the calmodulin 5' and 3' UTRs also induces the same hyperexcitable phenotype. Direct examination of transformed cells under voltage clamp reveals that two different Ca(2+)-activated K(+)-specific currents are reduced in amplitude. This reduction does not correlate with a deficit of PAK11 message, since RNA is clearly produced from the injected transgenes. Insertion of a single nucleotide at the start of the PAK11 ORF does not affect the RNA level but completely abolishes the phenotypic transformation. Thus, the reduction of K(+) currents by the expression of the K(+)-channel transgenes reported here is likely to be the consequence of a post translational event. The complexity of behavioral changes, possible mechanisms, and implications in Paramecium biology are discussed. PMID- 11729148 TI - Phenotypic and suppressor analysis of defecation in clk-1 mutants reveals that reaction to changes in temperature is an active process in Caenorhabditis elegans. AB - Mutations in the Caenorhabditis elegans maternal-effect gene clk-1 affect cellular, developmental, and behavioral timing. They result in a slowing of the cell cycle, embryonic and postembryonic development, reproduction, and aging, as well as of the defecation, swimming, and pharyngeal pumping cycles. Here, we analyze the defecation behavior in clk-1 mutants, phenotypically and genetically. When wild-type worms are grown at 20 degrees and shifted to a new temperature, the defecation cycle length is significantly affected by that new temperature. In contrast, we find that when clk-1 mutants are shifted, the defecation cycle length is unaffected by that new temperature. We carried out a screen for mutations that suppress the slow defecation phenotype at 20 degrees and identified two distinct classes of genes, which we call dsc for defecation suppressor of clk-1. Mutations in one class also restore the ability to react normally to changes in temperature, while mutations in the other class do not. Together, these results suggest that clk-1 is necessary for readjusting the defecation cycle length in response to changes in temperature. On the other hand, in the absence of clk-1 activity, we observe temperature compensation, a mechanism that maintains a constant defecation period in the face of changes in temperature. PMID- 11729149 TI - Caenorhabditis elegans MES-3 is a target of GLD-1 and functions epigenetically in germline development. AB - The maternal-effect sterile (MES) proteins are maternally supplied regulators of germline development in Caenorhabditis elegans. In the hermaphrodite progeny from mes mutant mothers, the germline dies during larval development. On the basis of the similarities of MES-2 and MES-6 to known transcriptional regulators and on the basis of the effects of mes mutations on transgene expression in the germline, the MES proteins are predicted to be transcriptional repressors. One of the MES proteins, MES-3, is a novel protein with no recognizable motifs. In this article we show that MES-3 is localized in the nuclei of embryos and germ cells, consistent with its predicted role in transcriptional regulation. Its distribution in the germline and in early embryos does not depend on the wild type functions of the other MES proteins. However, its nuclear localization in midstage embryos and its persistence in the primordial germ cells depend on wild type MES-2 and MES-6. These results are consistent with biochemical data showing that MES-2, MES-3, and MES-6 associate in a complex in embryos. The distribution of MES-3 in the adult germline is regulated by the translational repressor GLD-1: MES-3 is absent from the region of the germline where GLD-1 is known to be present, MES-3 is overexpressed in the germline of gld-1 mutants, and GLD-1 specifically binds the mes-3 3' untranslated region (3' UTR). Analysis of temperature-shifted mes-3(bn21ts) worms and embryos indicates that MES-3 function is required in the mother's germline and during embryogenesis to ensure subsequent normal germline development. We propose that MES-3 acts epigenetically to induce a germline state that is inherited through both meiosis and mitosis and that is essential for survival of the germline. PMID- 11729150 TI - Depletion of a novel SET-domain protein enhances the sterility of mes-3 and mes-4 mutants of Caenorhabditis elegans. AB - Four maternal-effect sterile genes, mes-2, mes-3, mes-4, and mes-6, are essential for germline development in Caenorhabditis elegans. Homozygous mes progeny from heterozygous mothers are themselves fertile but produce sterile progeny with underproliferated and degenerated germlines. All four mes genes encode chromatin associated proteins, two of which resemble known regulators of gene expression. To identify additional components in the MES pathway, we used RNA-mediated interference (RNAi) to test candidate genes for enhancement of the Mes mutant phenotype. Enhancement in this assay was induction of sterility a generation earlier, in the otherwise fertile homozygous progeny of heterozygous mothers, which previous results had suggested represent a sensitized genetic background. We tested seven genes predicted to encode regulators of chromatin organization for RNAi-induced enhancement of mes-3 sterility and identified one enhancer, called set-2 after the SET domain encoded by the gene. Depletion of SET-2 also enhances the sterile phenotype of mes-4 but not of mes-2 or mes-6. set-2 encodes two alternatively spliced transcripts, set-2(l) and set-2(s), both of which are enriched in the germline of adults. In the adult germline, SET-2(L) protein is localized in mitotic and mid-late-stage meiotic nuclei but is undetectable in early pachytene nuclei. SET-2(L) protein is localized in all nuclei of embryos. The localization of SET-2(L) does not depend on any of the four MES proteins, and none of the MES proteins depend on SET-2 for their normal localization. Our results suggest that SET-2 participates along with the MES proteins in promoting normal germline development. PMID- 11729152 TI - An analysis of polygenes affecting wing shape on chromosome 2 in Drosophila melanogaster. AB - Genetic effects on an index of wing shape on chromosome 2 of Drosophila melanogaster were mapped using isogenic recombinants with transposable element markers. At least 10 genes with small additive effects are dispersed evenly along the chromosome. Many interactions exist, with only small net effects in homozygous recombinants and little effect on phenotypic variance. Heterozygous chromosome segments show almost no dominance. Pleiotropic effects on leg shape are only minor. At first view, wing shape genes form a rather homogeneous class, but certain complexities remain unresolved. PMID- 11729151 TI - Drosophila-raf acts to elaborate dorsoventral pattern in the ectoderm of developing embryos. AB - In the early Drosophila embryo the activity of the EGF-receptor (Egfr) is required to instruct cells to adopt a ventral neuroectodermal fate. Using a gain of-function mutation we showed that D-raf acts to transmit this and other late acting embryonic Egfr signals. A novel role for D-raf was also identified in lateral cell development using partial loss-of-function D-raf mutations. Thus, we provide evidence that zygotic D-raf acts to specify cell fates in two distinct pathways that generate dorsoventral pattern within the ectoderm. These functional requirements for D-raf activity occur subsequent to its maternal role in organizing the anterioposterior axis. The consequences of eliminating key D-raf regulatory domains and specific serine residues in the transmission of Egfr and lateral epidermal signals were also addressed here. PMID- 11729153 TI - A large family of divergent Drosophila odorant-binding proteins expressed in gustatory and olfactory sensilla. AB - We identified a large family of putative odorant-binding protein (OBP) genes in the genome of Drosophila melanogaster. Some of these genes are present in large clusters in the genome. Most members are expressed in various taste organs, including gustatory sensilla in the labellum, the pharyngeal labral sense organ, dorsal and ventral cibarial organs, as well as taste bristles located on the wings and tarsi. Some of the gustatory OBPs are expressed exclusively in taste organs, but most are expressed in both olfactory and gustatory sensilla. Multiple binding proteins can be coexpressed in the same gustatory sensillum. Cells in the tarsi that express OBPs are required for normal chemosensation mediated through the leg, as ablation of these cells dramatically reduces the sensitivity of the proboscis extension reflex to sucrose. Finally, we show that OBP genes expressed in the pharyngeal taste sensilla are still expressed in the poxneuro genetic background while OBPs expressed in the labellum are not. These findings support a broad role for members of the OBP family in gustation and olfaction and suggest that poxneuro is required for cell fate determination of labellar but not pharyngeal taste organs. PMID- 11729154 TI - Analysis of corkscrew signaling in the Drosophila epidermal growth factor receptor pathway during myogenesis. AB - The Drosophila nonreceptor protein tyrosine phosphatase, Corkscrew (Csw), functions positively in multiple receptor tyrosine kinase (RTK) pathways, including signaling by the epidermal growth factor receptor (EGFR). Detailed phenotypic analyses of csw mutations have revealed that Csw activity is required in many of the same developmental processes that require EGFR function. However, it is still unclear where in the signaling hierarchy Csw functions relative to other proteins whose activities are also required downstream of the receptor. To address this issue, genetic interaction experiments were performed to place csw gene activity relative to the EGFR, spitz (spi), rhomboid (rho), daughter of sevenless (DOS), kinase-suppressor of ras (ksr), ras1, D-raf, pointed (pnt), and moleskin. We followed the EGFR-dependent formation of VA2 muscle precursor cells as a sensitive assay for these genetic interaction studies. First, we established that Csw has a positive function during mesoderm development. Second, we found that tissue-specific expression of a gain-of-function csw construct rescues loss of-function mutations in other positive signaling genes upstream of rolled (rl)/MAPK in the EGFR pathway. Third, we were able to infer levels of EGFR signaling in various mutant backgrounds during myogenesis. This work extends previous studies of Csw during Torso and Sevenless RTK signaling to include an in depth analysis of the role of Csw in the EGFR signaling pathway. PMID- 11729155 TI - The functions of the multiproduct and rapidly evolving dec-1 eggshell gene are conserved between evolutionarily distant species of Drosophila. AB - The Drosophila dec-1 gene encodes multiple proteins that are required for female fertility and proper eggshell morphogenesis. Genetic and immunolocalization data suggest that the different DEC-1 proteins are functionally distinct. To identify regions within the proteins with potential biological significance, we cloned and sequenced the D. yakuba and D. virilis dec-1 homologs. Interspecies comparisons of the predicted translation products revealed rapidly evolving sequences punctuated by blocks of conserved amino acids. Despite extensive amino acid variability, the proteins produced by the different dec-1 homologs were functionally interchangeable. The introduction of transgenes containing either the D. yakuba or the D. virilis dec-1 open reading frames into a D. melanogaster DEC-1 protein null mutant was sufficient to restore female fertility and wild type eggshell morphology. Normal expression and extracellular processing of the DEC-1 proteins was correlated with the phenotypic rescue. The nature of the conserved features highlighted by the evolutionary comparison and the molecular resemblance of some of these features to those found in other extracellular proteins suggests functional correlates for some of the multiple DEC-1 derivatives. PMID- 11729156 TI - Expanding the diversity of the IS630-Tc1-mariner superfamily: discovery of a unique DD37E transposon and reclassification of the DD37D and DD39D transposons. AB - A novel transposon named ITmD37E was discovered in a wide range of mosquito species. Sequence analysis of multiple copies in three Aedes species showed similar terminal inverted repeats and common putative TA target site duplications. The ITmD37E transposases contain a conserved DD37E catalytic motif, which is unique among reported transposons of the IS630-Tc1-mariner superfamily. Sequence comparisons and phylogenetic analyses suggest that ITmD37E forms a novel family distinct from the widely distributed Tc1 (DD34E), mariner (DD34D), and pogo (DDxD) families in the IS630-Tc1-mariner superfamily. The inclusion in the phylogenetic analysis of recently reported transposons and transposons uncovered in our database survey provided revisions to previous classifications and identified two additional families, ITmD37D and ITmD39D, which contain DD37D and DD39D motifs, respectively. The above expansion and reorganization may open the doors to the discovery of related transposons in a broad range of organisms and help illustrate the evolution and structure-function relationships among these distinct transposases in the IS630-Tc1-mariner superfamily. The presence of intact open reading frames and highly similar copies in some of the newly characterized transposons suggests recent transposition. Studies of these novel families may add to the limited repertoire of transgenesis and mutagenesis tools for a wide range of organisms, including the medically important mosquitoes. PMID- 11729157 TI - Drosophila rhino encodes a female-specific chromo-domain protein that affects chromosome structure and egg polarity. AB - Here we describe our analyses of Rhino, a novel member of the Heterochromatin Protein 1(HP1) subfamily of chromo box proteins. rhino (rhi) is expressed only in females and chiefly in the germline, thus providing a new tool to dissect the role of chromo-domain proteins in development. Mutations in rhi disrupt eggshell and embryonic patterning and arrest nurse cell nuclei during a stage-specific reorganization of their polyploid chromosomes, a mitotic-like state called the "five-blob" stage. These visible alterations in chromosome structure do not affect polarity by altering transcription of key patterning genes. Expression levels of gurken (grk), oskar (osk), bicoid (bcd), and decapentaplegic (dpp) transcripts are normal, with a slight delay in the appearance of bcd and dpp mRNAs. Mislocalization of grk and osk transcripts, however, suggests a defect in the microtubule reorganization that occurs during the middle stages of oogenesis and determines axial polarity. This defect likely results from aberrant Grk/Egfr signaling at earlier stages, since rhi mutations delay synthesis of Grk protein in germaria and early egg chambers. In addition, Grk protein accumulates in large, actin-caged vesicles near the endoplasmic reticulum of stages 6-10 egg chambers. We propose two hypotheses to explain these results. First, Rhi may play dual roles in oogenesis, independently regulating chromosome compaction in nurse cells at the end of the unique endoreplication cycle 5 and repressing transcription of genes that inhibit Grk synthesis. Thus, loss-of-function mutations arrest nurse cell chromosome reorganization at the five-blob stage and delay production or processing of Grk protein, leading to axial patterning defects. Second, Rhi may regulate chromosome compaction in both nurse cells and oocyte. Loss-of-function mutations block nurse cell nuclear transitions at the five-blob stage and activate checkpoint controls in the oocyte that arrest Grk synthesis and/or inhibit cytoskeletal functions. These functions may involve direct binding of Rhi to chromosomes or may involve indirect effects on pathways controlling these processes. PMID- 11729159 TI - Evidence for new alleles in the protein kinase adenosine monophosphate-activated gamma(3)-subunit gene associated with low glycogen content in pig skeletal muscle and improved meat quality. AB - Several quantitative trait loci (QTL) affecting muscle glycogen content and related traits were mapped to pig chromosome 15 using a three-generation intercross between Berkshire x Yorkshire pigs. On the basis of the QTL location the PRKAG3 (protein kinase, AMP-activated, gamma(3)-subunit) gene was considered to be a good candidate for the observed effects. Differences in the PRKAG3 gene sequences of the founder animals of the intercross were analyzed. The RN(-) mutation previously reported was not present in the cross but three missense substitutions and a polymorphic short interspersed element (SINE) were identified. To confirm the hypothesis that at least one of these mutations was associated with differences in meat quality, >1800 animals from several unrelated commercial lines were genotyped for the candidate substitutions and an association study was performed. The results demonstrate the presence of new economically important alleles of the PRKAG3 gene affecting the glycogen content in the muscle and the resulting meat quality. Haplotype analysis was shown to resolve the effects of PRKAG3 more clearly than analysis of individual polymorphisms. Because of their prevalence in the more common commercial breeds, the potential implications for the pig industry and consumers are considerably greater than the original discovery of the RN(-) mutation. Furthermore, these results illustrate that additional alleles of genes involved in major mutations may play a significant role in quantitative trait variation. PMID- 11729158 TI - The Trithorax-mimic allele of Enhancer of zeste renders active domains of target genes accessible to polycomb-group-dependent silencing in Drosophila melanogaster. AB - Two antagonistic groups of genes, the trithorax- and the Polycomb-group, are proposed to maintain the appropriate active or inactive state of homeotic genes set up earlier by transiently expressed segmentation genes. Although some details about the mechanism of maintenance are available, it is still unclear how the initially active or inactive chromatin domains are recognized by either the trithorax-group or the Polycomb-group proteins. We describe an unusual dominant allele of a Polycomb-group gene, Enhancer of zeste, which mimics the phenotype of loss-of-function mutations in trithorax-group genes. This mutation, named E(z)(Trithorax mimic) [E(z)(Trm)], contains a single-amino-acid substitution in the conserved SET domain. The strong dominant trithorax-like phenotypes elicited by this E(z) allele suggest that the mutated arginine-741 plays a critical role in distinguishing between active and inactive chromatin domains of the homeotic gene complexes. We have examined the modification of E(z)(Trm) phenotypes by mutant alleles of PcG and trxG genes and other mutations that alter the phosphorylation of nuclear proteins, covalent modifications of histones, or histone dosage. These data implicate some trxG genes in transcriptional repression as well as activation and provide genetic evidence for involvement of histone modifications in PcG/trxG-dependent transcriptional regulation. PMID- 11729160 TI - Pooling analysis of genetic data: the association of leptin receptor (LEPR) polymorphisms with variables related to human adiposity. AB - Analysis of raw pooled data from distinct studies of a single question generates a single statistical conclusion with greater power and precision than conventional metaanalysis based on within-study estimates. However, conducting analyses with pooled genetic data, in particular, is a daunting task that raises important statistical issues. In the process of analyzing data pooled from nine studies on the human leptin receptor (LEPR) gene for the association of three alleles (K109R, Q223R, and K656N) of LEPR with body mass index (BMI; kilograms divided by the square of the height in meters) and waist circumference (WC), we encountered the following methodological challenges: data on relatives, missing data, multivariate analysis, multiallele analysis at multiple loci, heterogeneity, and epistasis. We propose herein statistical methods and procedures to deal with such issues. With a total of 3263 related and unrelated subjects from diverse ethnic backgrounds such as African-American, Caucasian, Danish, Finnish, French-Canadian, and Nigerian, we tested effects of individual alleles; joint effects of alleles at multiple loci; epistatic effects among alleles at different loci; effect modification by age, sex, diabetes, and ethnicity; and pleiotropic genotype effects on BMI and WC. The statistical methodologies were applied, before and after multiple imputation of missing observations, to pooled data as well as to individual data sets for estimates from each study, the latter leading to a metaanalysis. The results from the metaanalysis and the pooling analysis showed that none of the effects were significant at the 0.05 level of significance. Heterogeneity tests showed that the variations of the nonsignificant effects are within the range of sampling variation. Although certain genotypic effects could be population specific, there was no statistically compelling evidence that any of the three LEPR alleles is associated with BMI or waist circumference in the general population. PMID- 11729162 TI - Codon usage bias covaries with expression breadth and the rate of synonymous evolution in humans, but this is not evidence for selection. AB - In numerous species, from bacteria to Drosophila, evidence suggests that selection acts even on synonymous codon usage: codon bias is greater in more abundantly expressed genes, the rate of synonymous evolution is lower in genes with greater codon bias, and there is consistency between genes in the same species in which codons are preferred. In contrast, in mammals, while nonequal use of alternative codons is observed, the bias is attributed to the background variance in nucleotide concentrations, reflected in the similar nucleotide composition of flanking noncoding and exonic third sites. However, a systematic examination of the covariants of codon usage controlling for background nucleotide content has yet to be performed. Here we present a new method to measure codon bias that corrects for background nucleotide content and apply this to 2396 human genes. Nearly all (99%) exhibit a higher amount of codon bias than expected by chance. The patterns associated with selectively driven codon bias are weakly recovered: Broadly expressed genes have a higher level of bias than do tissue-specific genes, the bias is higher for genes with lower rates of synonymous substitutions, and certain codons are repeatedly preferred. However, while these patterns are suggestive, the first two patterns appear to be methodological artifacts. The last pattern reflects in part biases in usage of nucleotide pairs. We conclude that we find no evidence for selection on codon usage in humans. PMID- 11729163 TI - Structural features and methylation patterns associated with paramutation at the r1 locus of Zea mays. AB - In paramutation, two alleles of a gene interact and, during the interaction, one of them becomes epigenetically silenced. The various paramutation systems that have been studied to date exhibit intriguing differences in the physical complexity of the loci involved. B and Pl alleles that participate in paramutation are simple, single genes, while the R haplotypes that participate in paramutation contain multiple gene copies and often include rearrangements. The number and arrangement of the sequences in particular complex R haplotypes have been correlated with paramutation behavior. Here, the physical structures of 28 additional haplotypes of R were examined. A specific set of physical features is associated with paramutability (the ability to be silenced). However, no physical features were strongly correlated with paramutagenicity (the ability to cause silencing) or neutrality (the inability to participate in paramutation). Instead, paramutagenic haplotypes were distinguished by high levels of cytosine methylation over certain regions of the genes while neutral haplotypes were distinguished by lack of C-methylation over these regions. These findings suggest that paramutability of r1 is determined by the genetic structure of particular haplotypes, while paramutagenicity is determined by the epigenetic state. PMID- 11729161 TI - Female meiosis drives karyotypic evolution in mammals. AB - Speciation is often accompanied by changes in chromosomal number or form even though such changes significantly reduce the fertility of hybrid intermediates. We have addressed this evolutionary paradox by expanding the principle that nonrandom segregation of chromosomes takes place whenever human or mouse females are heterozygous carriers of Robertsonian translocations, a common form of chromosome rearrangement in mammals. Our analysis of 1170 mammalian karyotypes provides strong evidence that karyotypic evolution is driven by nonrandom segregation during female meiosis. The pertinent variable in this form of meiotic drive is the presence of differing numbers of centromeres on paired homologous chromosomes. This situation is encountered in all heterozygous carriers of Robertsonian translocations. Whenever paired chromosomes have different numbers of centromeres, the inherent asymmetry of female meiosis and the polarity of the meiotic spindle dictate that the partner with the greater number of centromeres will attach preferentially to the pole that is most efficient at capturing centromeres. This mechanism explains how chromosomal variants become fixed in populations, as well as why closely related species often appear to have evolved by directional adjustment of the karyotype toward or away from a particular chromosome form. If differences in the ability of particular DNA sequences or chromosomal regions to function as centromeres are also considered, nonrandom segregation is likely to affect karyotype evolution across a very broad phylogenetic range. PMID- 11729164 TI - Inbreeding depression in small populations of self-incompatible plants. AB - Self-incompatibility (SI) is a widespread mechanism that prevents inbreeding in flowering plants. In many species, SI is controlled by a single locus (the S locus) where numerous alleles are maintained by negative frequency-dependent selection. Inbreeding depression, the decline in fitness of selfed individuals compared to outcrossed ones, is an essential factor in the evolution of SI systems. Conversely, breeding systems influence levels of inbreeding depression. Little is known about the joint effect of SI and drift on inbreeding depression. Here we studied, using a two-locus model, the effect of SI (frequency-dependent selection) on a locus subject to recurrent deleterious mutations causing inbreeding depression. Simulations were performed to assess the effect of population size and linkage between the two loci on the level of inbreeding depression and genetic load. We show that the sheltering of deleterious alleles linked to the S locus strengthens inbreeding depression in small populations. We discuss the implications of our results for the evolution of SI systems. PMID- 11729165 TI - An integrated map of Arabidopsis thaliana for functional analysis of its genome sequence. AB - The genome of the model plant species Arabidopsis thaliana has recently been sequenced. To accelerate its current genome research, we developed a whole genome, BAC/BIBAC-based, integrated physical, genetic, and sequence map of the A. thaliana ecotype Columbia. This new map was constructed from the clones of a new plant-transformation-competent BIBAC library and is integrated with the existing sequence map. The clones were restriction fingerprinted by DNA sequencing gel based electrophoresis, assembled into contigs, and anchored to an existing genetic map. The map consists of 194 BAC/BIBAC contigs, spanning 126 Mb of the 130-Mb Arabidopsis genome. A total of 120 contigs, spanning 114 Mb, were anchored to the chromosomes of Arabidopsis. Accuracy of the integrated map was verified using the existing physical and sequence maps and numerous DNA markers. Integration of the new map with the sequence map has enabled gap closure of the sequence map and will facilitate functional analysis of the genome sequence. The method used here has been demonstrated to be sufficient for whole-genome physical mapping from large-insert random bacterial clones and thus is applicable to rapid development of whole-genome physical maps for other species. PMID- 11729166 TI - Sex determination in the androdioecious plant Datisca glomerata and its dioecious sister species D. cannabina. AB - Datisca glomerata is an androdioecious plant species containing male and hermaphroditic individuals. Molecular markers and crossing data suggest that, in both D. glomerata and its dioecious sister species D. cannabina, sex is determined by a single nuclear locus, at which maleness is dominant. Supporting this conclusion, an amplified fragment length polymorphism (AFLP) is heterozygous in males and homozygous recessive in hermaphrodites in three populations of the androdioecious species. Additionally, hermaphrodite x male crosses produced 1:1 sex ratios, while hermaphrodite x hermaphrodite crosses produced almost entirely hermaphroditic offspring. No perfectly sex-linked marker was found in the dioecious species, but all markers associated with sex mapped to a single linkage group and were heterozygous in the male parent. There was no sex-ratio heterogeneity among crosses within D. cannabina collections, but males from one collection produced highly biased sex ratios (94% females), suggesting that there may be sex-linked meiotic drive or a cytoplasmic sex-ratio factor. Interspecific crosses produced only male and female offspring, but no hermaphrodites, suggesting that hermaphroditism is recessive to femaleness. This comparative approach suggests that the hermaphrodite form arose in a dioecious population from a recessive mutation that allowed females to produce pollen. PMID- 11729168 TI - Characterization of mutations that feminize gametophytes of the fern Ceratopteris. AB - Gametophytes of the fern Ceratopteris are either male or hermaphroditic. Their sex is epigenetically determined by the pheromone antheridiogen, which is secreted by the hermaphrodite and induces male and represses female development in other young, sexually undetermined gametophytes. To understand how antheridiogen represses the development of female traits at the genetic level, 16 new mutations that feminize the gametophyte in the presence of antheridiogen were identified and characterized. Seven are very tightly linked to the FEM1 locus previously described. Nine others define another locus named NOTCHLESS1 (NOT1), as several of the not1 mutants lack a meristem notch. Some not1 mutations also affect sporophyte development only when homozygous, indicating that the not1 mutations are recessive and that NOT1 is also required for normal sporophyte development. The epistatic interactions among FEM1, NOT1, and other sex determining genes are described. This information was used to expand the genetic model of the sex-determining pathway in Ceratopteris. On the basis of this model, we can say that the presence of antheridiogen leads to the activation of the FEM1 gene, which not only promotes the differentiation of male traits, but also represses female development by activating the NOT1 gene. NOT1 represses the TRA genes necessary for the development of female traits in the gametophyte. PMID- 11729167 TI - The distribution of early recombination nodules on zygotene bivalents from plants. AB - Early recombination nodules (ENs) are protein complexes approximately 100 nm in diameter that are associated with forming synaptonemal complexes (SCs) during leptotene and zygotene of meiosis. Although their functions are not yet clear, ENs may have roles in synapsis and recombination. Here we report on the frequency and distribution of ENs in zygotene SC spreads from six plant species that include one lower vascular plant, two dicots, and three monocots. For each species, the number of ENs per unit length is higher for SC segments than for (asynapsed) axial elements (AEs). In addition, EN number is strongly correlated with SC segment length. There are statistically significant differences in EN frequencies on SCs between species, but these differences are not related to genome size, number of chromosomes, or phylogenetic class. There is no difference in the frequency of ENs per unit length of SC from early to late zygotene. The distribution of distances between adjacent ENs on SC segments is random for all six species, but ENs are found at synaptic forks more often than expected for a random distribution of ENs on SCs. From these observations, we conclude that in plants: (1) some ENs bind to AEs prior to synapsis, (2) most ENs bind to forming SCs at synaptic forks, and (3) ENs do not bind to already formed SCs. PMID- 11729169 TI - A chromosomal region promoting outcrossing in a conifer. AB - Prefertilization mechanisms influencing selfing rates are thought to be absent in conifers. Outcrossing in conifers is promoted via an embryo-lethal system, but the genetic mechanism is poorly understood. This study is the first experimental profile of the genetic mechanism promoting outcrossing in conifers. Molecular dissection of a Pinus taeda L. selfed pedigree detected a chromosomal region identified as PtTX3020-RPtest9. Within this region, a semilethal factor was tightly linked (r = 0.0076) to a polymorphic expressed sequence tag (EST). The linkage group flanking the lethal factor showed strong heterozygote advantage. Using genotypic frequencies for the linkage group, three hypotheses about the semilethal factor could be tested: (1) the presence of a balanced lethal system, i.e., a lethal factor present in each of the two marker intervals; (2) gametic selection operative prior to fertilization; and (3) a stage-specific lethal factor. Selection acted via the embryo-lethal system. No support for a genetic mechanism operating prior to fertilization was found. The semilethal factor exerted no effect after embryo maturity. The genetic mechanism promoting outcrossing in P. taeda L. appears to have a balancing selection system due to either pseudo-overdominance or true overdominance. PMID- 11729170 TI - Probabilistic prediction of unknown metabolic and signal-transduction networks. AB - Regulatory networks provide control over complex cell behavior in all kingdoms of life. Here we describe a statistical model, based on representing proteins as collections of domains or motifs, which predicts unknown molecular interactions within these biological networks. Using known protein-protein interactions of Saccharomyces cerevisiae as training data, we were able to predict the links within this network with only 7% false-negative and 10% false-positive error rates. We also use Markov chain Monte Carlo simulation for the prediction of networks with maximum probability under our model. This model can be applied across species, where interaction data from one (or several) species can be used to infer interactions in another. In addition, the model is extensible and can be analogously applied to other molecular data (e.g., DNA sequences). PMID- 11729171 TI - Estimating recombination rates from population genetic data. AB - We introduce a new method for estimating recombination rates from population genetic data. The method uses a computationally intensive statistical procedure (importance sampling) to calculate the likelihood under a coalescent-based model. Detailed comparisons of the new algorithm with two existing methods (the importance sampling method of Griffiths and Marjoram and the MCMC method of Kuhner and colleagues) show it to be substantially more efficient. (The improvement over the existing importance sampling scheme is typically by four orders of magnitude.) The existing approaches not infrequently led to misleading results on the problems we investigated. We also performed a simulation study to look at the properties of the maximum-likelihood estimator of the recombination rate and its robustness to misspecification of the demographic model. PMID- 11729172 TI - Population admixture may appear to mask, change or reverse genetic effects of genes underlying complex traits. AB - Association studies using random population samples are increasingly being applied in the identification and inference of genetic effects of genes underlying complex traits. It is well recognized that population admixture may yield false-positive identification of genetic effects for complex traits. However, it is less well appreciated that population admixture can appear to mask, change, or reverse true genetic effects for genes underlying complex traits. By employing a simple population genetics model, we explore the effects and the conditions of population admixture in masking, changing, or even reversing true genetic effects of genes underlying complex traits. PMID- 11729173 TI - Mapping quantitative trait Loci using generalized estimating equations. AB - A number of statistical methods are now available to map quantitative trait loci (QTL) relative to markers. However, no existing methodology can simultaneously map QTL for multiple nonnormal traits. In this article we rectify this deficiency by developing a QTL-mapping approach based on generalized estimating equations (GEE). Simulation experiments are used to illustrate the application of the GEE based approach. PMID- 11729174 TI - A multivalent pairing model of linkage analysis in autotetraploids. AB - Polyploidy has been recognized as an important step in the evolutionary diversification of flowering plants and may have a significant impact on plant breeding. Statistical analyses for linkage mapping in polyploid species can be difficult due to considerable complexities in polysomic inheritance. In this article, we develop a novel statistical method for linkage analysis of polymorphic markers in a full-sib family of autotetraploids. This method is established on multivalent pairings of homologous chromosomes at meiosis and can provide a simultaneous maximum-likelihood estimation of the double reduction frequencies of and recombination fraction between two markers. The EM algorithm is implemented to provide a tractable way for estimating relative proportions of different modes of gamete formation that generate identical gamete genotypes due to multivalent pairings. Extensive simulation studies were performed to demonstrate the statistical properties of this method. The implications of the new method for understanding the genome structure and organization of polyploid species are discussed. PMID- 11729175 TI - Bayesian methods for quantitative trait loci mapping based on model selection: approximate analysis using the Bayesian information criterion. AB - We describe an approximate method for the analysis of quantitative trait loci (QTL) based on model selection from multiple regression models with trait values regressed on marker genotypes, using a modification of the easily calculated Bayesian information criterion to estimate the posterior probability of models with various subsets of markers as variables. The BIC-delta criterion, with the parameter delta increasing the penalty for additional variables in a model, is further modified to incorporate prior information, and missing values are handled by multiple imputation. Marginal probabilities for model sizes are calculated, and the posterior probability of nonzero model size is interpreted as the posterior probability of existence of a QTL linked to one or more markers. The method is demonstrated on analysis of associations between wood density and markers on two linkage groups in Pinus radiata. Selection bias, which is the bias that results from using the same data to both select the variables in a model and estimate the coefficients, is shown to be a problem for commonly used non Bayesian methods for QTL mapping, which do not average over alternative possible models that are consistent with the data. PMID- 11729176 TI - Estimating the total number of alleles using a sample coverage method. AB - Previously reported methods for estimating the number of different alleles at a single locus in a population have not described a useful general result. Using the number of alleles observed in a sample gives an underestimate for the true number of alleles. The similar problem of estimating the number of species in a population was first investigated in 1943. In this article we use the sample coverage method proposed by Chao and Lee in 1992 to estimate the number of alleles in a population when there are unequal allele frequencies. Simulation studies under the recurrent mutation model show that, for reasonable sample sizes, a significantly better estimate of the true number can be obtained than that using only the observed alleles. Results under the stepwise mutation model and infinite-allele model are presented. Possible applications include improving the characterization of the prior distribution for the allele frequencies, adjusting the estimates of genetic diversity, and estimating the range of microsatellite alleles. PMID- 11729177 TI - On prediction of genetic values in marker-assisted selection. AB - We suggest a new approximation for the prediction of genetic values in marker assisted selection. The new approximation is compared to the standard approach. It is shown that the new approach will often provide substantially better prediction of genetic values; furthermore the new approximation avoids some of the known statistical problems of the standard approach. The advantages of the new approach are illustrated by a simulation study in which the new approximation outperforms both the standard approach and phenotypic selection. PMID- 11729178 TI - Study of amide-proton exchange of Escherichia coli melibiose permease by attenuated total reflection-Fourier transform infrared spectroscopy: evidence of structure modulation by substrate binding. AB - The accessibility of Escherichia coli melibiose permease to aqueous solvent was studied following hydrogen-deuterium exchange kinetics monitored by attenuated total reflection-Fourier transform infrared spectroscopy under four distinct conditions where MelB forms different complexes with its substrates (H(+), Na(+), melibiose). Analysis of the amide II band upon (2)H(2)O exposure discloses a significant sugar protection of the protein against aqueous solvent, resulting in an 8% less exchange of the corresponding H(+)*melibiose*MelB complex compared with the protein in the absence of sugar. Investigation of the amide I exchange reveals clear substrate effects on beta-sheet accessibility, with the complex H(+)*melibiose*MelB being the most protected state against exchange, followed by Na(+)*melibiose*MelB. Although of smaller magnitude, similar changes in alpha helices plus non-ordered structures are detected. Finally, no differences are observed when analyzing reverse turn structures. The results suggest that sugar binding induces a remarkable compactness of the carrier's structure, affecting mainly beta-sheet domains of the transporter, which, according to secondary structure predictions, may include cytoplasmic loops 4-5 and 10-11. A possible catalytic role of these two loops in the functioning of MelB is hypothesized. PMID- 11729179 TI - Localization of endothelial nitric-oxide synthase phosphorylated on serine 1179 and nitric oxide in Golgi and plasma membrane defines the existence of two pools of active enzyme. AB - The subcellular localization of endothelial nitric-oxide synthase (eNOS) is critical for optimal coupling of extracellular stimulation to nitric oxide production. Because eNOS is activated by Akt-dependent phosphorylation to produce nitric oxide (NO), we determined the subcellular distribution of eNOS phosphorylated on serine 1179 using a variety of methodologies. Based on sucrose gradient fractionation, phosphorylated-eNOS (P-eNOS) was found in both caveolin-1 enriched membranes and intracellular domains. Co-transfection of eNOS with Akt and stimulation of endothelial cells with vascular endothelial growth factor (VEGF) increased the ratio of P-eNOS to total eNOS but did not change the relative intracellular distribution between these domains. The proper localization of eNOS to intracellular membranes was required for agonist dependent phosphorylation on serine 1179, since VEGF did not increase eNOS phosphorylation in cells transfected with a non-acylated, mistargeted form of eNOS. Confocal imaging of P-eNOS and total eNOS pools demonstrated co localization in the Golgi region and plasmalemma of transfected cells and native endothelial cells. Finally, VEGF stimulated a large increase in NO localized in both the perinuclear region and the plasma membrane of endothelial cells. Thus, activated, phosphorylated eNOS resides in two cellular compartments and both pools are VEGF-regulated to produce NO. PMID- 11729180 TI - A major Echinococcus multilocularis antigen is a mucin-type glycoprotein. AB - The metacestode of Echinococcus multilocularis is surrounded by a carbohydrate rich laminated layer, which plays a key role in the establishment of the infection in the mammalian host. A major component of the laminated layer is an antigen referred to as Em2(G11). This highly species-specific antigen has been used for serodiagnoses of alveolar echinococcosis and is suggested to contain carbohydrates as major constituents. The results of this work have shown that immunoaffinity-purified Em2(G11) subjected to size-exclusion chromatography eluted mainly in the void volume, indicating a high molecular weight structure of this antigen. Amino acid analysis revealed a large proportion of threonine and proline residues in Em2(G11). The carbohydrate moiety of the antigen was found to be composed of galactose, N-acetylgalactosamine, and N-acetylglucosamine with a ratio of 2.4:1.0:0.5 as determined by gas-chromatography/mass spectrometry. An isotope tag was introduced to the beta-eliminated glycans, and an integrated mass spectrometric O-glycan profiling and sequencing approach was employed to obtain detailed sequence and linkage information of the unseparated glycoform pool. Novel glycoforms containing mucin-type core Gal1-3GalNAc and branched core structures attached to both serine and threonine residues are described. The data presented reveal that the Em2(G11) antigen is a mucin-type glycosylated protein. PMID- 11729181 TI - Phorbol 12-myristate 13-acetate inhibits death receptor-mediated apoptosis in Jurkat cells by disrupting recruitment of Fas-associated polypeptide with death domain. AB - Regulation of death receptor-mediated apoptosis is incompletely understood. Previous studies have demonstrated that phorbol 12-myristate 13-acetate (PMA), a protein kinase C activator, inhibits Fas (CD95)-mediated apoptosis in Jurkat (type II) cells but not SKW6.4 (type I) cells. In this study, we demonstrated that PMA also protects Jurkat cells from apoptosis induced by tumor necrosis factor-alpha and the tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL). Interestingly, PMA failed to protect Jurkat cells from apoptosis induced by other agents, including etoposide, camptothecin, and gamma irradiation. Analysis of the initial events induced by agonistic anti-Fas antibodies revealed that PMA inhibited Fas binding to Fas-associated polypeptide with death domain (FADD) in Jurkat cells but not in SKW6.4 cells. Although the protein kinase inhibitor bisindoylmaleimide VIII increased apoptosis induced by agonistic anti-Fas antibody, tumor necrosis factor-alpha, and TRAIL, these effects were not observed with the protein kinase C inhibitor H7 and were not associated with increased FADD recruitment to Fas. These results indicate that PMA inhibits death signaling induced by a number of discrete receptors and suggest that the effects are mediated at the level of receptor-mediated adaptor molecule recruitment. PMID- 11729182 TI - Chronic leptin administration decreases fatty acid uptake and fatty acid transporters in rat skeletal muscle. AB - Chronic leptin administration reduces triacylglycerol content in skeletal muscle. We hypothesized that chronic leptin treatment, within physiologic limits, would reduce the fatty acid uptake capacity of red and white skeletal muscle due to a reduction in transport protein expression (fatty acid translocase (FAT/CD36) and plasma membrane-associated fatty acid-binding protein (FABPpm)) at the plasma membrane. Female Sprague-Dawley rats were infused for 2 weeks with leptin (0.5 mg/kg/day) using subcutaneously implanted miniosmotic pumps. Control and pair-fed animals received saline-filled implants. Leptin levels were significantly elevated (approximately 4-fold; p < 0.001) in treated animals, whereas pair-fed treated animals had reduced serum leptin levels (approximately -2-fold; p < 0.01) relative to controls. Palmitate transport rates into giant sarcolemmal vesicles were reduced following leptin treatment in both red (-45%) and white (-84%) skeletal muscle compared with control and pair-fed animals (p < 0.05). Leptin treatment reduced FAT mRNA (red, -70%, p < 0.001; white, -48%, p < 0.01) and FAT/CD36 protein expression (red, -32%; p < 0.05) in whole muscle homogenates, whereas FABPpm mRNA and protein expression were unaltered. However, in leptin treated animals plasma membrane fractions of both FAT/CD36 and FABPpm protein expression were significantly reduced in red (-28 and -34%, respectively) and white (-44 and -56%, respectively) muscles (p < 0.05). Across all experimental treatments and muscles, palmitate uptake by giant sarcolemmal vesicles was highly correlated with the plasma membrane FAT/CD36 protein (r = 0.88, p < 0.01) and plasma membrane FABPpm protein (r = 0.94, p < 0.01). These studies provide the first evidence that protein-mediated long chain fatty acid transport is subject to long term regulation by leptin. PMID- 11729183 TI - Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts. AB - Ribosomal L10-L7/L12 protein complex and L11 bind to a highly conserved RNA region around position 1070 in domain II of 23 S rRNA and constitute a part of the GTPase-associated center in Escherichia coli ribosomes. We replaced these ribosomal proteins in vitro with the rat counterparts P0-P1/P2 complex and RL12, and tested them for ribosomal activities. The core 50 S subunit lacking the proteins on the 1070 RNA domain was prepared under gentle conditions from a mutant deficient in ribosomal protein L11. The rat proteins bound to the core 50 S subunit through their interactions with the 1070 RNA domain. The resultant hybrid ribosome was insensitive to thiostrepton and showed poly(U)-programmed polyphenylalanine synthesis dependent on the actions of both eukaryotic elongation factors 1alpha (eEF-1alpha) and 2 (eEF-2) but not of the prokaryotic equivalent factors EF-Tu and EF-G. The results from replacement of either the L10 L7/L12 complex or L11 with rat protein showed that the P0-P1/P2 complex, and not RL12, was responsible for the specificity of the eukaryotic ribosomes to eukaryotic elongation factors and for the accompanying GTPase activity. The presence of either E. coli L11 or rat RL12 considerably stimulated the polyphenylalanine synthesis by the hybrid ribosome, suggesting that L11/RL12 proteins play an important role in post-GTPase events of translation elongation. PMID- 11729184 TI - Association of the signaling adaptor FRS2 with fibroblast growth factor receptor 1 (Fgfr1) is mediated by alternative splicing of the juxtamembrane domain. AB - Fibroblast growth factor receptors (FGFRs) are a family of transmembrane tyrosine kinases involved in signaling via interactions with the family of fibroblast growth factors. Alternative splicing of the juxtamembrane region of FGFR1-3 leads to the inclusion or exclusion of two amino acids, valine and threonine, the VT site. The presence or absence of VT (VT+ or VT-, respectively) affects the signaling potential of the receptor. The VT+ receptor isoform is required for Erk2 phosphorylation, a component of the mitogen-activated protein kinase signaling pathway. FRS2 is an adaptor protein that links FGFRs to the mitogen activated protein kinase signaling pathway. FRS2 interacts with a region of the juxtamembrane domain of FGFR1 that includes the alternatively spliced VT site. We investigated the interaction of FRS2 with murine Fgfr1 juxtamembrane domain. We showed the alternatively spliced VT motif, at the juxtamembrane domain of Fgfr1 is required for FRS2 interaction with Fgfr1. Activation of signaling pathways from FRS2 is likely to be regulated by controlling the Fgfr1/FRS2 interaction through alternative splicing of the VT motif of Fgfr1. PMID- 11729185 TI - PTEN protects p53 from Mdm2 and sensitizes cancer cells to chemotherapy. AB - The PTEN tumor suppressor protein inhibits phosphatidylinositol 3-kinase (PI3K)/Akt signaling that promotes translocation of Mdm2 into the nucleus. When restricted to the cytoplasm, Mdm2 is degraded. The ability of PTEN to inhibit the nuclear entry of Mdm2 increases the cellular content and transactivation of the p53 tumor suppressor protein. Retroviral transduction of PTEN into U87MG (PTEN null) glioblastoma cells increases p53 activity and expression of p53 target genes and induces cell cycle arrest. U87MG/PTEN glioblastoma cells are more sensitive than U87MG/PTEN null cells to death induced by etoposide, a chemotherapeutic agent that induces DNA damage. Previously, tumor suppressor proteins have been supposed to act individually to suppress cancers. Our results establish a direct connection between the activities of two major tumor suppressors and show that they act together to respond to stresses and malignancies. PTEN protects p53 from survival signals, permitting p53 to function as a guardian of the genome. By virtue of its capacity to protect p53, PTEN can sensitize tumor cells to chemotherapy that relies on p53 activity. p53 induces PTEN gene expression, and here it is shown that PTEN protects p53, indicating that a positive feedback loop may amplify the cellular response to stress, damage, and cancer. PMID- 11729186 TI - Constitutive internalization of constitutively active agiotensin II AT(1A) receptor mutants is blocked by inverse agonists. AB - As constitutively active mutants (CAMs) mimic an active conformation, they can be used to characterize the process of G protein-coupled receptor activation. Here, we used CAMs to study the link between activation and internalization of the angiotensin II AT(1A) receptor. The cellular localization of fluorescently tagged N111A, I245T, and L305Q mutants was determined by confocal microscopy. In the absence of ligand, CAMs were mostly located in intracellular vesicles, whereas the wild-type AT(1A) was found at the cell surface. After 2 h incubation with inverse agonist, losartan, CAMs were translocated to the plasma membrane. Similar observations were made in H295, a human adrenocortical cell line which expresses physiologically the AT(1) receptor. This phenomenon, which was not dependent on protein synthesis and the pharmacology and kinetics of which were similar to the recycling of the wild-type receptor, was called "externalization". After externalization and losartan removal, the L305Q CAM underwent rapid ligand independent endocytosis, with the same kinetics and temperature sensitivity as the angiotensin II-induced internalization of the wild-type AT(1A). Moreover, the addition of a second mutation known to block internalization (Delta 329 truncation) prevented intracellular localization of the CAM. These data show that AT(1A) CAMs are constitutively and permanently internalized and recycled. This mechanism is different from the down-regulation observed for CAMs of other G protein-coupled receptors and thus defines a new paradigm for the cellular regulation of CAMs. PMID- 11729187 TI - Many type IIs restriction endonucleases interact with two recognition sites before cleaving DNA. AB - Type IIs restriction endonucleases recognize asymmetric DNA sequences and cleave both DNA strands at fixed positions, typically several base pairs away from the recognition site. These enzymes are generally monomers that transiently associate to form dimers to cleave both strands. Their reactions could involve bridging interactions between two copies of their recognition sequence. To examine this possibility, several type IIs enzymes were tested against substrates with either one or two target sites. Some of the enzymes cleaved the DNA with two target sites at the same rate as that with one site, but most cut their two-site substrate more rapidly than the one-site DNA. In some cases, the two sites were cut sequentially, at rates that were equal to each other but that exceeded the rate on the one-site DNA. In another case, the DNA with two sites was cleaved rapidly at one site, but the residual site was cleaved at a much slower rate. In a further example, the two sites were cleaved concertedly to give directly the final products cut at both sites. Many type IIs enzymes thus interact with two copies of their recognition sequence before cleaving DNA, although via several different mechanisms. PMID- 11729188 TI - The type IIs restriction endonuclease BspMI is a tetramer that acts concertedly at two copies of an asymmetric DNA sequence. AB - Type IIs endonucleases recognize asymmetric DNA sequences and cleave both strands at fixed positions downstream of the sequence. Many type IIs enzymes, including BspMI, cleave substrates with two sites more rapidly than those with one site. They usually act sequentially on DNA with two sites, but BspMI converted such a substrate directly to the final products cut at both sites. The BspMI endonuclease was found to be a tetramer, in contrast to the monomeric structures for many type IIs enzymes. No change in subunit association occurred during the BspMI reaction. Plasmids with two BspMI sites were cleaved in cis, in reactions spanning sites in the same DNA, even when the sites were separated by just 38 bp. Plasmids with one BspMI site were cleaved in trans, with the enzyme bridging sites in separate DNA molecules: these slow reactions could be accelerated by adding a second DNA with the recognition sequence. Thus, whereas many type IIs enzymes dimerize before cleaving DNA, a process facilitated by two recognition sites in cis, the BspMI tetramer binds two copies of its recognition sequence before cleaving the DNA in both strands at both sites. PMID- 11729189 TI - Inactivation of the amidotransferase activity of carbamoyl phosphate synthetase by the antibiotic acivicin. AB - Carbamoyl phosphate synthetase (CPS) from Escherichia coli catalyzes the formation of carbamoyl phosphate from 2 mol of ATP, bicarbonate, and glutamine. CPS was inactivated by the glutamine analog, acivicin. In the presence of ATP and bicarbonate the second-order rate constant for the inactivation of the glutamine dependent activities was 4.0 x 10(4) m(-1) s(-1). In the absence of ATP and bicarbonate the second-order rate constant for inactivation of CPS was reduced by a factor of 200. The enzyme was protected against inactivation by the inclusion of glutamine in the reaction mixture. The ammonia-dependent activities were unaffected by the incubation of CPS with acivicin. These results are consistent with the covalent labeling of the glutamine-binding site located within the small amidotransferase subunit. The binding of ATP and bicarbonate to the large subunit of CPS must also induce a conformational change within the amidotransferase domain of the small subunit that enhances the nucleophilic character of the thiol group required for glutamine hydrolysis. The acivicin-inhibited enzyme was crystallized, and the three-dimensional structure was determined by x-ray diffraction techniques. The thiol group of Cys-269 was covalently attached to the dihydroisoxazole ring of acivicin with the displacement of a chloride ion. PMID- 11729190 TI - Shear stress stimulates phosphorylation of endothelial nitric-oxide synthase at Ser1179 by Akt-independent mechanisms: role of protein kinase A. AB - Recently, we have shown that shear stress stimulates NO(*) production by the protein kinase B/Akt (Akt)-dependent mechanisms in bovine aortic endothelial cells (BAEC) (Go, Y. M., Boo, Y. C., Park, H., Maland, M. C., Patel, R., Pritchard, K. A., Jr., Fujio, Y., Walsh, K., Darley-Usmar, V., and Jo, H. (2001) J. Appl. Physiol. 91, 1574-1581). Akt has been believed to regulate shear dependent production of NO(*) by directly phosphorylating endothelial nitric oxide synthase (eNOS) at the Ser(1179) residue (eNOS-S(1179)), but a critical evaluation using specific inhibitors or dominant negative mutants (Akt(AA) or Akt(AAA)) has not been reported. In addition, other kinases, including protein kinase A (PKA) and AMP kinase have also shown to phosphorylate eNOS-S(1179). Here, we show that shear-dependent phosphorylation of eNOS-S(1179) is mediated by an Akt-independent, but a PKA-dependent, mechanism. Expression of Akt(AA) or Akt(AAA) in BAEC by using recombinant adenoviral constructs inhibited phosphorylation of eNOS-S(1179) if cells were stimulated by vascular endothelial growth factor (VEGF), but not by shear stress. As shown before, expression of Akt(AA) inhibited shear-dependent NO(*) production, suggesting that Akt is still an important regulator in NO production. Further studies showed that a selective inhibitor of PKA, H89, inhibited shear-dependent phosphorylation of eNOS-S(1179) and NO(*) production. In contrast, H89 did not inhibit phosphorylation of eNOS S(1179) induced by expressing a constitutively active Akt mutant (Akt(Myr)) in BAEC, showing that the inhibitor did not affect the Akt pathway. 8-Bromo-cAMP alone phosphorylated eNOS-S(1179) within 5 min without activating Akt, in an H89 sensitive manner. Collectively, these results demonstrate that shear stimulates phosphorylation of eNOS-S(1179) in a PKA-dependent, but Aktindependent manner, whereas the NO(*) production is regulated by the mechanisms dependent on both PKA and Akt. A coordinated interaction between Akt and PKA may be an important mechanism by which eNOS activity is regulated in response to physiological stimuli such as shear stress. PMID- 11729191 TI - Water-assisted dual mode cofactor recognition by HhaI DNA methyltransferase. AB - Energetically competent binary recognition of the cofactor S-adenosyl-L methionine (AdoMet) and the product S-adenosyl-L-homocysteine (AdoHcy) by the DNA (cytosine C-5) methyltransferase (M.HhaI) is demonstrated herein. Titration calorimetry reveals a dual mode, involving a primary dominant exothermic reaction followed by a weaker endothermic one, for the recognition of AdoMet and AdoHcy by M.HhaI. Conservation of the bimodal recognition in W41I and W41Y mutants of M.HhaI excludes the cation-pi interaction between the methylsulfonium group of AdoMet and the pi face of the Trp(41) indole ring from a role in its origin. Small magnitude of temperature-independent heat capacity changes upon AdoMet or AdoHcy binding by M.HhaI preclude appreciable conformational alterations in the reacting species. Coupled osmotic-calorimetric analyses of AdoMet and AdoHcy binding by M.HhaI indicate that a net uptake of nearly eight and 10 water molecules, respectively, assists their primary recognition. A change in water activity at constant temperature and pH is sufficient to engender and conserve enthalpy-entropy compensation, consistent with a true osmotic effect. The results implicate solvent reorganization in providing the major contribution to the origin of this isoequilibrium phenomenon in AdoMet and AdoHcy recognition by M.HhaI. The observations provide unequivocal evidence for the binding of AdoMet as well as AdoHcy to M.HhaI in solution state. Isotope partitioning analysis and preincubation studies favor a random mechanism for M.HhaI-catalyzed reaction. Taken together, the results clearly resolve the issue of cofactor recognition by free M.HhaI, specifically in the absence of DNA, leading to the formation of an energetically and catalytically competent binary complex. PMID- 11729192 TI - Protein kinase C signaling regulates ZO-1 translocation and increased paracellular flux of T84 colonocytes exposed to Clostridium difficile toxin A. AB - Clostridium difficile toxin A increases paracellular permeability in colonic epithelial T84 cells by mechanisms involving RhoA glucosylation and actin depolymerization. However, we previously observed that toxin A-mediated decline in transepithelial electrical resistance preceded changes in cell morphology and tight junction ultrastructure (Hecht, G., Pothoulakis, C., LaMont, J. T., and Madara, J. L. (1988) J. Clin. Invest. 82, 1516-1524). Recent studies also showed that C. difficile toxins induce early cellular responses, including activation of mitogen-activated protein kinases, generation of reactive oxygen metabolites, and calcium influx. The aim of this study was to investigate whether toxin A-induced early cellular responses contribute to the permeability changes. We found that toxin A stimulated the activities of membrane and cytosolic protein kinase Calpha (PKCalpha) and cytosolic PKCbeta. A specific PKCalpha/beta antagonist (myristoylated PKCalpha/beta peptide) blocked toxin A-mediated RhoA glucosylation. Furthermore, decreased transepithelial electrical resistance and increased translocation of ZO-1 from tight junction occurred within 2-3 h of toxin A exposure and were also inhibited by PKCalpha/beta antagonist. During this time period, toxin exposure did not induce translocation of ZO-2, dephosphorylation or translocation of occludin, or cell rounding. Our data indicate that PKC signaling regulates toxin A-mediated paracellular permeability changes and ZO-1 translocation. PMID- 11729193 TI - Interaction of CED-6/GULP, an adapter protein involved in engulfment of apoptotic cells with CED-1 and CD91/low density lipoprotein receptor-related protein (LRP). AB - The prompt clearance of cells undergoing apoptosis is critical during embryonic development, normal tissue turnover, as well as inflammation and autoimmunity. The molecular details of the engulfment of apoptotic cells are not fully understood. ced-6 and its human homologue gulp, encode an adapter protein, whose function in engulfment is highly evolutionarily conserved; however, the upstream and downstream components of CED-6 mediated signaling are not known. Recently, ced-1 has been shown to encode a transmembrane protein on phagocytic cells, with two functional sequence motifs in its cytoplasmic tail that are important for engulfment. In this study, using a combination of biochemical approaches and yeast two-hybrid analysis, we present evidence for a physical interaction between GULP/CED-6 and one of the two motifs (NPXY motif) in the cytoplasmic tail of CED 1. The phosphotyrosine binding domain of GULP was necessary and sufficient for this interaction. Since the precise mammalian homologue of CED-1 is not known, we undertook a database search for human proteins that contain the motifs shown to be important for CED-1 function and identified CD91/LRP (low density lipoprotein receptor-related protein) as one candidate. Interestingly, recent studies have also identified CD91/LRP as a receptor involved in the phagocytosis of apoptotic cells in mammals. The GULP phosphotyrosine binding domain was able to specifically interact with one specific NPXY motif in the CD91 cytoplasmic tail. During these studies we have also identified the mouse GULP sequence. These studies suggest a physical link between CED-1 or CD91/LRP and the adapter protein CED-6/GULP during engulfment of apoptotic cells and further elucidate the pathway suggested by the genetic studies. PMID- 11729194 TI - Pnk1, a DNA kinase/phosphatase required for normal response to DNA damage by gamma-radiation or camptothecin in Schizosaccharomyces pombe. AB - We report the characterization of Pnk1, a 45-kDa homolog of the human polynucleotide kinase PNKP in Schizosaccharomyces pombe. Recombinant Pnk1 like human PNKP exhibits both 5'-DNA kinase and 3'-DNA phosphatase activities in vitro. Furthermore, we detected 3'-DNA phosphatase activity with a single stranded substrate in extracts from wild-type yeast, but no activity was detected in pnk1delta strains. We have shown that GFP-tagged Pnk1 like mammalian PNKP localizes to the nucleus. Deletion of pnk1 does not affect cell growth under normal conditions but results in significant hypersensitivity to gamma-radiation or camptothecin, an inhibitor of topoisomerase I, suggesting that Pnk1 plays an important role in the repair of DNA strand breaks produced by these agents. The pnk1 deletion mutants were not hypersensitive to ethyl methanesulfonate, methyl methanesulfonate, or 4-nitroquinoline N-oxide. Expression of human PNKP in pnk1delta cells restores resistance to gamma-radiation or camptothecin, suggesting that the functions of yeast Pnk1 and human PNKP have been conserved. PMID- 11729195 TI - The p40phox and p47phox PX domains of NADPH oxidase target cell membranes via direct and indirect recruitment by phosphoinositides. AB - The Phox homology (PX) domain has recently been reported to bind to phosphoinositides, and some PX domains can localize to endosomes in vivo. Here we show data to support the conclusion that the p40(phox) PX domain binds to phosphatidylinositol 3-phosphate specifically in vitro and localizes to endosomes in intact cells. In addition, its Y59A/L65Q mutant, which has decreased affinity for phosphatidylinositol 3-phosphate in vitro, fails to target EGFP-p40-PX to endosomes. However, unlike published results, we find that the p47(phox) PX domain weakly binds to many phosphoinositides in vitro showing slightly higher affinity for phosphatidylinositol 3,4,5-trisphosphate. Moreover, we show for the first time that upon insulin-like growth factor-1 stimulation of COS cells, the p47(phox) PX domain is localized to the plasma membrane, and this subcellular localization is dependent on PI 3-kinase activity. Unexpectedly, its R42Q mutant that loses in vitro phosphoinositide-binding ability can still target EGFP-p47-PX to the plasma membrane. Our data suggest that the translocation of p47(phox) PX domain to the plasma membrane does involve 3'-phosphoinositide(s) in the process, but the phosphoinositide-binding of p47(phox) PX domain is not sufficient to recruit it to the plasma membrane. Therefore, the p40(phox) and p47(phox) PX domains can target subcellular membranes via direct or indirect recruitment by phosphoinositides, while both are under the control of phosphatidylinositol 3 kinase activity. PMID- 11729196 TI - Mutations in the carboxyl-terminal domain of phospholipase C-beta 1 delineate the dimer interface and a potential Galphaq interaction site. AB - The carboxyl-terminal domain of phospholipase C-beta is required for its stimulation by Galpha(q) and for its Galpha(q)-specific GTPase-activating protein (GAP) activity. We subjected this domain to a combination of deletion and alanine/glycine scanning mutagenesis to detect mutations that would inhibit either responsiveness to G(q) or G(q) GAP activity. Most mutations that altered either response or GAP activity diminished both in parallel. Many of these mutations map at the interface at which the carboxyl-terminal domain was recently shown to form a dimer (Singer, A. U., et al. (2001) Nat. Struct. Biol., 9, 32 36). Most others clustered in an area that is a plausible Galpha(q) binding site. In addition, one mutation that differentially inhibited GAP activity relative to responsiveness to Galpha(q) mapped in this region at a location modeled to be in close contact with the switch II region of Galpha(q). This is the site at which RGS proteins are thought to exert their GAP activity. Last, a deletion mutation differentially inhibited the response of phospholipase C-beta1 to Galpha(q) without blocking GAP activity. Its location in the molecule suggests that moving the attachment point of the catalytic domain can disrupt its ability to be activated by Galpha(q). PMID- 11729197 TI - The X-ray crystal structure and putative ligand-derived peptide binding properties of gamma-aminobutyric acid receptor type A receptor-associated protein. AB - The gamma-aminobutyric acid receptor type A (GABA(A)) receptor-associated protein (GABARAP) has been reported to mediate the interaction between the GABA(A) receptor and microtubules. We present the three-dimensional structure of GABARAP obtained by x-ray diffraction at 1.75 A resolution. The structure was determined by molecular replacement using the structure of the homologous protein GATE-16. NMR spectroscopy of isotope-labeled GABARAP showed the structure in solution to be compatible with the overall fold but showed evidence of conformation heterogeneity that is not apparent in the crystal structure. We assessed the binding of GABARAP to peptides derived from reported binding partner proteins, including the M3-M4 loop of the gamma2 subunit of the GABA(A) receptor and the acidic carboxyl-terminal tails of human alpha- and beta-tubulin. There is a small area of concentrated positive charge on one surface of GABARAP, which we found interacts weakly with all peptides tested, but we found no evidence for specific binding to the proposed physiological target peptides. These results are compatible with a more general role in membrane targeting and transportation for the GABARAP family of proteins. PMID- 11729198 TI - A post-Amadori inhibitor pyridoxamine also inhibits chemical modification of proteins by scavenging carbonyl intermediates of carbohydrate and lipid degradation. AB - Reactive carbonyl compounds are formed during autoxidation of carbohydrates and peroxidation of lipids. These compounds are intermediates in the formation of advanced glycation end products (AGE) and advanced lipoxidation end products (ALE) in tissue proteins during aging and in chronic disease. We studied the reaction of carbonyl compounds glyoxal (GO) and glycolaldehyde (GLA) with pyridoxamine (PM), a potent post-Amadori inhibitor of AGE formation in vitro and of development of renal and retinal pathology in diabetic animals. PM reacted rapidly with GO and GLA in neutral, aqueous buffer, forming a Schiff base intermediate that cyclized to a hemiaminal adduct by intramolecular reaction with the phenolic hydroxyl group of PM. This bicyclic intermediate dimerized to form a five-ring compound with a central piperazine ring, which was characterized by electrospray ionization-liquid chromatography/mass spectrometry, NMR, and x-ray crystallography. PM also inhibited the modification of lysine residues and loss of enzymatic activity of RNase in the presence of GO and GLA and inhibited formation of the AGE/ALE N(epsilon)-(carboxymethyl)lysine during reaction of GO and GLA with bovine serum albumin. Our data suggest that the AGE/ALE inhibitory activity and the therapeutic effects of PM observed in diabetic animal models depend, at least in part, on its ability to trap reactive carbonyl intermediates in AGE/ALE formation, thereby inhibiting the chemical modification of tissue proteins. PMID- 11729199 TI - Densin-180 interacts with delta-catenin/neural plakophilin-related armadillo repeat protein at synapses. AB - Densin-180, a protein purified from the postsynaptic density fraction of the rat forebrain, is the founding member of a newly described family of proteins termed the LAP (leucine-rich repeats and PSD-95/Dlg-A/ZO-1 (PDZ) domains) family that plays essential roles in establishment of cell polarity. To identify Densin-180 binding proteins, we screened a yeast two-hybrid library using the carboxyl terminal fragment of Densin-180 containing PDZ domain as bait, and we isolated delta-catenin/neural plakophilin-related armadillo repeat protein (NPRAP) as a Densin-180-interacting protein. delta-catenin/NPRAP, a member of the armadillo repeat family, is a nervous system-specific adherens junction protein originally discovered as an interactor with presenilin-1, a protein involved in Alzheimer's disease. Densin-180 PDZ domain binds the COOH terminus of delta-catenin/NPRAP containing the PDZ domain-binding sequence. Endogenous Densin-180 was co immunoprecipitated with delta-catenin/NPRAP and N-cadherin. Although Densin-180 was reported to be a transmembrane protein, Densin-180 was not accessible to surface biotinylation in dissociated hippocampal neurons; hence Densin-180 may be a cytosolic protein. Densin-180 co-localized with delta-catenin/NPRAP at synapses in delta-catenin/NPRAP and may be involved in organization of the synaptic cell cell junction through interaction with the delta-catenin/NPRAP-N-cadherin complex. PMID- 11729200 TI - Role of neutrophil NADPH oxidase in the mechanism of tumor necrosis factor-alpha induced NF-kappa B activation and intercellular adhesion molecule-1 expression in endothelial cells. AB - In this study, we explored a novel function of polymorphonuclear neutrophils (PMN) NAD(P)H oxidase in the mechanism of tumor necrosis factor-alpha (TNFalpha) induced NF-kappaB activation and intercellular adhesion molecule-1 (ICAM-1) expression in endothelial cells. Studies were made in mice lacking the p47(phox) subunit of NAD(P)H oxidase as well as in cultured mouse lung vascular endothelial cells (MLVEC) from these mice. In response to TNFalpha challenge, NF-kappaB activation and ICAM-1 expression were significantly attenuated in lungs of p47(phox)(-/-) mice as compared with wild-type (WT) mice. The attenuated NF kappaB activation in p47(phox)(-/-) mice was secondary to inhibition of NIK activity and subsequent IkappaBalpha degradation. Induction of neutropenia using anti-PMN serum prevented the initial TNFalpha-induced NF-kappaB activation and ICAM-1 expression in WT mice, indicating the involvement of PMN NAD(P)H oxidase in signaling these responses. Moreover, the responses were restored upon repletion with PMN obtained from WT mice but not with PMN from p47(phox)(-/-) mice. These findings were recapitulated in MLVEC co-cultured with PMN, suggesting that NF-kappaB activation and resultant ICAM-1 expression in endothelial cells occurred secondary to oxidants generated by the PMN NAD(P)H oxidase complex. The functional relevance of the PMN NAD(P)H oxidase in mediating TNFalpha-induced ICAM-1-dependent endothelial adhesivity was evident by markedly reduced adhesion of p47(phox)(-/-) PMN in co-culture experiments. Thus, oxidant signaling by the PMN NAD(P)H oxidase complex is an important determinant of TNFalpha-induced NF kappaB activation and ICAM-1 expression in endothelial cells. PMID- 11729201 TI - Agonist-induced internalization of the platelet-activating factor receptor is dependent on arrestins but independent of G-protein activation. Role of the C terminus and the (D/N)PXXY motif. AB - As with most G-protein-coupled receptors, repeated agonist stimulation of the platelet-activating factor receptor (PAFR) results in its desensitization, sequestration, and internalization. In this report, we show that agonist-induced PAFR internalization is independent of G-protein activation but is dependent on arrestins and involves the interaction of arrestins with a limited region of the PAFR C terminus. In cotransfected COS-7 cells, both arrestin-2 and arrestin-3 could be coimmunoprecipitated with PAFR, and agonist stimulation of PAFR induced the translocation of both arrestin-2 and arrestin-3. Furthermore, coexpression of arrestin-2 with PAFR potentiated receptor internalization, whereas agonist induced PAFR internalization was inhibited by a dominant negative mutant of arrestin-2. The coexpression of a minigene encoding the C-terminal segment of the receptor abolished PAF-induced arrestin translocation and inhibited PAFR internalization. Using C terminus deletion mutants, we determined that the association of arrestin-2 with the receptor was dependent on the region between threonine 305 and valine 330 because arrestin-2 could be immunoprecipitated with the mutant PAFRstop330 but not PAFRstop305. Consistently, stop330 could mediate agonist-induced arrestin-2 translocation, whereas stop305 could not. Two other deletion mutants with slightly longer regions of the C terminus, PAFRstop311 and PAFRstop317, also failed to induce arrestin-2 translocation. Finally, the PAFR mutant Y293A, containing a single substitution in the putative internalization motif DPXXY in the seventh transmembrane domain (which we had shown to be able to internalize but not to couple to G-proteins) could efficiently induce arrestin translocation. Taken together, our results indicate that ligand-induced PAFR internalization is dependent on arrestins, that PAFR can associate with both arrestin-2 and -3, and that their translocation involves interaction with the region of residues 318-330 in the PAFR C terminus but is independent of G-protein activation. PMID- 11729202 TI - Prevalent loss of mitotic spindle checkpoint in adult T-cell leukemia confers resistance to microtubule inhibitors. AB - Human T-cell leukemia virus type I (HTLV-I) is the causative agent for adult T cell leukemia (ATL). Molecularly, ATL cells have extensive aneugenic abnormalities that occur, at least in part, from cell cycle dysregulation by the HTLV-I-encoded Tax oncoprotein. Here, we compared six HTLV-I-transformed cells to Jurkat and primary peripheral blood mononuclear cells (PBMC) in their responses to treatment with microtubule inhibitors. We found that both Jurkat and PBMCs arrested efficiently in mitosis when treated with nocodazole. By contrast, all six HTLV-I cells failed to arrest comparably in mitosis, suggesting that ATL cells have a defect in the mitotic spindle assembly checkpoint. Mechanistically, we observed that in HTLV-I Tax-expressing cells human spindle assembly checkpoint factors hsMAD1 and hsMAD2 were mislocated from the nucleus to the cytoplasm. This altered localization of hsMAD1 and hsMAD2 correlated with loss of mitotic checkpoint function and chemoresistance to microtubule inhibitors. PMID- 11729203 TI - Metabolic engineering in yeast demonstrates that S-adenosylmethionine controls flux through the methylenetetrahydrofolate reductase reaction in vivo. AB - One-carbon flux into methionine and S-adenosylmethionine (AdoMet) is thought to be controlled at the methylenetetrahydrofolate reductase (MTHFR) step. Mammalian MTHFRs are inhibited by AdoMet in vitro, and it has been proposed that methyl group biogenesis is regulated in vivo by this feedback loop. In this work, we used metabolic engineering in the yeast Saccharomyces cerevisiae to test this hypothesis. Like mammalian MTHFRs, the yeast MTHFR encoded by the MET13 gene is NADPH-dependent and is inhibited by AdoMet in vitro. This contrasts with plant MTHFRs, which are NADH-dependent and AdoMet-insensitive. To manipulate flux through the MTHFR reaction in yeast, the chromosomal copy of MET13 was replaced by an Arabidopsis MTHFR cDNA (AtMTHFR-1) or by a chimeric sequence (Chimera-1) comprising the yeast N-terminal domain and the AtMTHFR-1 C-terminal domain. Chimera-1 used both NADH and NADPH and was insensitive to AdoMet, supporting the view that the C-terminal domain is responsible for AdoMet inhibition. Engineered yeast expressing Chimera-1 accumulated 140-fold more AdoMet and 7-fold more methionine than did the wild-type and grew normally. Yeast expressing AtMTHFR-1 accumulated 8-fold more AdoMet. This is the first in vivo evidence that the AdoMet sensitivity and pyridine nucleotide preference of MTHFR control methylneogenesis. (13)C labeling data indicated that glycine cleavage becomes a more prominent source of one-carbon units when Chimera-1 is expressed. Possibly related to this shift in one-carbon fluxes, total folate levels are doubled in yeast cells expressing Chimera-1. PMID- 11729204 TI - A single, bi-functional aquaglyceroporin in blood-stage Plasmodium falciparum malaria parasites. AB - The malaria parasite Plasmodium falciparum faces drastic osmotic changes during kidney passages and is engaged in the massive biosynthesis of glycerolipids during its development in the blood-stage. We identified a single aquaglyceroporin (PfAQP) in the nearly finished genome of P. falciparum with highest similarity to the Escherichia coli glycerol facilitator (50.4%), but both canonical Asn-Pro-Ala (NPA) motifs in the pore region are changed to Asn-Leu-Ala (NLA) and Asn-Pro-Ser (NPS), respectively. Expression in Xenopus oocytes renders them highly permeable for both water and glycerol. Sugar alcohols up to five carbons and urea pass the pore. Mutation analyses of the NLA/NPS motifs showed their structural importance, but the symmetrical pore properties were maintained. PfAQP is expressed in blood-stage parasites throughout the development from rings via trophozoites to schizonts and is localized to the parasite but not to the erythrocyte cytoplasm or membrane. Its unique bi-functionality indicates functions in the protection from osmotic stress and efficiently provides access to the serum glycerol pool for the use in ATP generation and primarily in the phospholipid synthesis. PMID- 11729205 TI - Activation of zeaxanthin is an obligatory event in the regulation of photosynthetic light harvesting. AB - By dynamic changes in protein structure and function, the photosynthetic membranes of plants are able to regulate the partitioning of absorbed light energy between utilization in photosynthesis and photoprotective non-radiative dissipation of the excess energy. This process is controlled by features of the intact membrane, the transmembrane pH gradient, the organization of the photosystem II antenna proteins and the reversible binding of a specific carotenoid, zeaxanthin. Resonance Raman spectroscopy has been applied for the first time to wild type and mutant Arabidopsis leaves and to intact thylakoid membranes to investigate the nature of the absorption changes obligatorily associated with the energy dissipation process. The observed changes in the carotenoid Resonance Raman spectrum proved that zeaxanthin was involved and indicated a dramatic change in zeaxanthin environment that specifically alters the pigment configuration and red-shifts the absorption spectrum. This activation of zeaxanthin is a key event in the regulation of light harvesting. PMID- 11729206 TI - Structural basis for nucleotide-dependent regulation of membrane-associated guanylate kinase-like domains. AB - CASK is a member of the membrane-associated guanylate kinases (MAGUK) homologs, a family of proteins that scaffold protein complexes at particular regions of the plasma membrane by utilizing multiple protein-binding domains. The GK domain of MAGUKs, which shares high similarity in amino acid sequence with yeast guanylate kinase (yGMPK), is the least characterized MAGUK domain both in structure and function. In addition to its scaffolding function, the GK domain of hCASK has been shown to be involved in transcription regulation. Here we report the crystal structure of the GK domain of human CASK (hCASK-GK) at 1.3-A resolution. The structure rationalizes the inability of the GK domain to catalyze phosphoryl transfer and strongly supports its new function as a protein-binding module. Comparison of the hCASK-GK structure with the available crystal structures of yGMPK provides insight into possible conformational changes that occur in hCASK upon GMP binding. These conformational changes may act to regulate hCASK-GK function in a nucleotide-dependent manner. PMID- 11729207 TI - Identification and functional characterization of distinct critically important bone morphogenetic protein-specific response elements in the Id1 promoter. AB - Transforming growth factor-beta (TGF-beta) family members, which include bone morphogenetic proteins (BMPs) and TGF-betas, elicit their cellular effects by activating specific Smad proteins, which control the transcription of target genes. BMPs and TGF-betas have overlapping as well as specific effects on mesenchymal cell differentiation for which the mechanisms are incompletely understood. Here we report that Id1, a dominant negative inhibitor of basic helix loop-helix proteins, is a direct target gene for BMP. BMP, but not TGF-beta, strongly activates the Id1 promoter in an Smad-dependent manner. We identified two BMP-responsive regions in the mouse Id1 promoter, which contain three distinct sequence elements; one region contains two Smad binding elements (SBEs), and the other region contains a GGCGCC palindromic sequence flanked by two CAGC and two CGCC motifs. Whereas SBEs and GGCGCC sequence are critically important, the CAGC and CGCC motifs are needed for efficient BMP-induced Id1 promoter activation. Smads are part of nuclear transcription factor complexes that specifically bind to SBEs and GGCGCC sequence in response to BMP but not TGF beta. Multimerization of the all three distinct sequence motifs is needed to generate a highly sensitive and BMP/Smad-dependent specific enhancer. Our results provide important new insights into how the BMP/Smad pathway can specifically activate target genes. PMID- 11729209 TI - ACP Best Practice No 167: the laboratory diagnosis of urinary tract infection. AB - Urinary tract infection is common, and it is not surprising that urine specimens make up a large proportion of those samples submitted to the routine diagnostic laboratory. Many of these specimens will show no evidence of infection and several methods can be used to screen out negative samples. Those that grow bacteria need to be carefully assessed to quantify the degree of bacteriuria and hence clinical relevance. To influence treatment, a final report should be produced within 24 hours of specimen receipt, with turnaround times continuously monitored. Much work needs to be done to determine the cost effectiveness involved in processing urine specimens and the evidence base for the final report provided. PMID- 11729210 TI - Light chain amyloidosis of the urinary bladder. A site restricted deposition of an externally produced immunoglobulin. AB - AIMS: To identify the amyloid protein in a patient with amyloidosis localised to the urinary bladder, and to see whether subtyping of the protein by sequence analysis increases the understanding of the selection of the urinary bladder as the site of amyloid deposition. METHODS: A patient with gross haematuria and a congophilic mass in his urinary bladder was evaluated further. Characterisation of the amyloid protein was performed using conventional histological and immunohistochemical methods. Determination of the N-terminal amino acid sequence of the amyloid protein was performed using protein sequencers. RESULTS: The patient's history, physical examination, and laboratory evaluation excluded the involvement of other organs, justifying a diagnosis of amyloidosis localised to the urinary bladder. Histological and immunological studies showed that the amyloid protein deposited in the urinary bladder of the patient was probably of the amyloid light chain type. No plasma cells or lymphocytes were seen in sections of the urinary bladder and lower ureter adjacent to the amyloid deposits. Molecular analysis showed the sequence NFMLTQPHSISGSPG, which assigned the amyloid protein to either the Vlambda(I) or the Vlambda(VI) immunoglobulin (Ig) light chain families. CONCLUSIONS: The findings suggest that the amyloid protein in this patient originated outside the urinary bladder. The heterogeneity of the Ig proteins in known cases of amyloidosis of the lower urinary tract suggests that the amino acid residues, which determine the Vlambda subtyping, have no major role in restricting the deposited protein to the urinary bladder. PMID- 11729211 TI - Evaluation of different continuous cell lines in the isolation of mumps virus by the shell vial method from clinical samples. AB - AIMS: To compare prospectively the efficacy of the Vero, LLC-MK2, MDCK, Hep-2, and MRC-5 cell lines in the isolation of the mumps virus from clinical samples by means of the shell vial method. METHODS: During an epidemic outbreak of parotiditis 48 clinical samples (saliva swabs and CSF) were studied. Two vials of the Vero, LLC-MK2, MDCK, MRC-5, and Hep-2 cell lines were inoculated with 0.2 ml of the samples by the shell vial assay. The vials were incubated at 36 degrees C for two and five days. The vials were then fixed with acetone at -20 degrees C for 10 minutes and stained by a monoclonal antibody against mumps virus by means of an indirect immunofluorescence assay. RESULTS: The mumps virus was isolated from 36 samples. The Vero and LLC-MK2 cell lines showed a 100% isolation capacity, MDCK showed 77.7%, MRC-5 showed 44.4%, and Hep-2 showed 22.2%. The Vero and LLC-MK2 lines were significantly different to the other cell lines (p < 0.001). The sensitivity for the Vero and LLC-MK2 lines at two and five days of incubation was identical (100%). The values obtained in the study of the quantitative isolation capacity (positive isolation with > 5 infectious foci) were 94.4% for Vero, 97.2% for LLC-MK2, 5.5% for MDCK, 5.5% for Hep-2, and 0% for MRC-5. CONCLUSIONS: The Vero and LLC-MK2 cell lines are equally efficient at two and five days incubation for the isolation of the mumps virus from clinical samples, and the use of the shell vial method considerably shortens the time of aetiological diagnosis with higher specificity. PMID- 11729208 TI - Neuroblastoma tumour genetics: clinical and biological aspects. AB - Neuroblastoma tumour cells show complex combinations of acquired genetic aberrations, including ploidy changes, deletions of chromosome arms 1p and 11q, amplification of the MYCN oncogene, and-most frequently-gains of chromosome arm 17q. Despite intensive investigation, the fundamental role of these features in neuroblastoma initiation and progression remains to be understood. Nonetheless, great progress has been made in relating tumour genetic abnormalities to tumour behaviour and to clinical outcome; indeed, neuroblastoma provides a paradigm for the clinical importance of tumour genetic abnormalities. Knowledge of MYCN status is increasingly being used in treatment decisions for individual children, and the clinical value of 1p and 17q data as adjuncts or refinements in risk stratification is under active investigation. Reliable detection of these molecular cytogenetic features should be regarded as mandatory for all new cases at presentation. PMID- 11729212 TI - The expression of antiapoptotic proteins Bcl-2, Bcl-X(L), and Mcl-1 in benign, dysplastic, and malignant biliary epithelium. AB - AIM: Cholangiocarcinoma can be cured by surgery, but only in a minority of cases. The activation of apoptosis is a major mode of action of chemotherapy and radiotherapy, which have limited benefit in the treatment of cholangiocarcinoma. The antiapoptotic members of the Bcl-2 protein family (Bcl-2, Bcl-X(L), and Mcl 1) are important inhibitors of apoptosis, but have not been investigated extensively in cholangiocarcinoma. METHODS: The expression of Bcl-2, Bcl-X(L), and Mcl-1 was investigated in normal biliary epithelium (17), biliary dysplasia (three), and invasive cholangiocarcinoma (51), in addition to three human cholangiocarcinoma cell lines, by immunohistochemistry and immunofluorescence. RESULTS: The expression of Bcl-2 was not detected in normal or malignant biliary tissue. In contrast, granular cytoplasmic Bcl-X(L) and Mcl-1 staining was found in 60-100% of cells in all normal, dysplastic, and malignant specimens, including the human cell lines examined in this study. CONCLUSION: These findings indicate that Mcl-1 and Bcl-X(L), but not Bcl-2, are involved in the survival of normal and neoplastic cells in the biliary tree. By prolonging survival through blocking apoptosis, these proteins might be reducing the efficacy of cytotoxic anticancer treatments in cholangiocarcinoma. PMID- 11729213 TI - MUC1 (EMA) is preferentially expressed by ALK positive anaplastic large cell lymphoma, in the normally glycosylated or only partly hypoglycosylated form. AB - AIMS: To investigate whether MUC1 mucin, a high molecular weight transmembrane glycoprotein, also known as epithelial membrane antigen (EMA), differs in its expression and degree of glycosylation between anaplastic large cell lymphoma (ALCL) and classic Hodgkin's disease (HD), and whether MUC1 immunostaining can be used to differentiate between CD30 positive large cell lymphomas. METHODS/RESULTS: Using five different monoclonal antibodies (E29/anti-EMA, DF3, 139H2, VU-4H5, and SM3) that distinguish between various MUC1 glycoforms, high MUC1 expression (50-95% of tumour cells positive) was found in 13 of 17 anaplastic lymphoma kinase (ALK) positive systemic nodal ALCLs, and in one of 20 cases of classic HD. Scattered or focal staining (< 25% of tumour cells) was seen in two additional ALK positive systemic ALCLs, two additional classic HD cases, and in three of 20 cases of ALK negative systemic nodal ALCL. Primary cutaneous ALCL showed no staining with the anti-MUC1 antibodies. Antibodies detecting hypoglycosylated MUC1 were found to be absent in all lymphomas (SM3) or present in only six of 15 ALK positive ALCLs (VU-4H5). CONCLUSIONS: MUC1 is preferentially expressed by a subtype of systemic nodal ALCL, characterised by ALK expression, but is found in only a few cases of classic HD and ALK negative ALCL. Therefore, although MUC1 could be used in a panel of markers for CD30 positive lymphomas, it is probably not a valuable tool to differentiate between ALK negative CD30 positive large cell lymphomas. Finally, the degree of MUC1 glycosylation in lymphomas is relatively high, compared with the aberrant hypoglycosylation found in adenocarcinomas. PMID- 11729214 TI - The prognostic relevance of angiogenesis and mast cells in squamous cell carcinoma of the oesophagus. AB - AIMS: Angiogenesis, an important prognostic factor in several tumours, is a complex event mediated by angiogenic factors released from cancer cells and host immune cells. Among the host immune cells, a role has been implicated for mast cells in tumour progression via promoting angiogenesis. Data have been recorded that indicate a correlation between intratumoral neovascularisation, as assessed by microvessel density (MVD), and prognosis in squamous cell carcinoma (SCC) of the oesophagus. However, a correlation between mast cell density (MCD) and either prognosis or angiogenesis has not been delineated yet in this disease. The aim of this study was to investigate the prognostic value of MVD and MCD in SCC of the oesophagus. The correlation between MVD and MCD was also evaluated. METHODS: MVD and MCD were investigated in tumour specimens from 53 patients diagnosed with SCC of the oesophagus. Intratumoral microvessels were stained with anti-CD34 antibody and mast cells with toluidine blue before being measured by light microscopy. RESULTS: Both MVD and MCD were associated with the depth of wall invasion, lymph node metastasis, and tumour progression (stage). A significant correlation was noted between MVD and MCD values (r = 0.72). The prognosis was significantly worse in patients with high MVD (> or = 92) and high MCD (> or = 18) values. Multivariate analysis indicated that MVD and stage were independent predictors of survival. CONCLUSIONS: These findings support the suggestion that MVD is a reliable prognostic marker in SCC of the oesophagus. Moreover, MCD may have a role in the angiogenesis of these tumours and might be responsible for their aggressive behaviour. PMID- 11729215 TI - Immunohistochemical analysis of E-cadherin, alpha-catenin, beta-catenin, gamma catenin, and neural cell adhesion molecule (NCAM) in chordoma. AB - AIMS: The epithelioid features seen in chordoma are unique among mesenchymal tumours. However, no detailed analysis regarding cell-cell communication has been conducted in this epithelioid tumour. The aims of this study were to investigate cell-cell communication in chordoma. METHODS: By means of immunohistochemical techniques that incorporated a panel of monoclonal antibodies against cell adhesion molecules (CAMs), including E-cadherin, alpha-catenin, beta-catenin, gamma-catenin, and neural cell adhesion molecule (NCAM), the expression of CAMs was studied in 15 specimens of chordoma and eight specimens of chondrosarcoma. RESULTS: Most chordoma specimens showed some positive immunoreactivity for all the CAMs examined. For the various CAMs investigated, between two and five cases showed diffuse immunoreactions, indicating well preserved expression. Well preserved expression of all the CAMs examined was limited to only one case, thus indicating that the expression of CAMs was decreased in most of the chordoma specimens; however, no significant correlation was found between the decreased expression of CAMs and the histological grade of malignancy, cellular growth pattern, or clinical parameters in chordoma. In chondrosarcoma, only a few specimens showed positive immunoreactivity for CAMs and the expression of E cadherin, beta-catenin, gamma-catenin, and NCAM was seen more frequently in the chordoma specimens than in the chondrosarcoma specimens. CONCLUSIONS: These results suggest that the expression of CAMs is associated with the formation and maintenance of chordoma tissue architecture, just as it is in other epithelial tumours or normal tissue. Immunohistochemistry for CAMs was found to be of diagnostic value for discriminating chordoma from chondrosarcoma, and these markers could be used along with the cytokeratins, which are already used for this purpose. PMID- 11729216 TI - Comparison of core oestrogen receptor (ER) assay with excised tumour: intratumoral distribution of ER in breast carcinoma. AB - AIMS: The use of the H score (involving the assessment of intensity and distribution of positivity) on sections stained for the oestrogen receptor (ER) by immunocytochemistry (ICC) allows different samples to be compared and detailed correlations to be made between hormone receptor expression and morphology. This study assessed the reliability of core biopsy in predicting ER expression in the same tumour excised later at treatment. The distribution of ER within excised tumours was investigated. METHODS: The distribution of ER positivity was investigated in 51 diagnostic core biopsies and across the diameter of 51 subsequently excised tumours in a field by field (magnification, x40; field diameter, 0.4 mm) assessment using the semiquantitive H scoring system. RESULTS: The ER H score in diagnostic core biopsy was significantly higher (p = 0.05, paired rank test; overall mean, 130; n = 51) than the mean in the corresponding excised tumour (mean, 110; n = 51). There was a significant downward trend in ER positivity from the periphery of tumours towards the centre (p = 0.001). The reduction of ER positivity was 6 H score units (2%)/mm. If core biopsies were orientated with the tumour edge at one end no change in ER positivity with field number along the length of the core could be demonstrated. CONCLUSIONS: ER estimation in core biopsies correlated well with expression in tumours but ER expression was higher in the core biopsies than in the excised tumours. ER expression was higher at the periphery of tumours than at the centre. The higher ER expression in cores may reflect the higher chance of sampling the peripheral part of a tumour using a needle core. PMID- 11729217 TI - Expression of prolactin receptors in normal, benign, and malignant breast tissue: an immunohistological study. AB - AIMS: Prolactin plays an important role in the proliferation and differentiation of normal breast epithelium, and possibly in the development of breast carcinoma. The effects of prolactin are mediated by its receptor; thus, alteration in the expression of this receptor could be important in studying the biology of breast cancer. This investigation was aimed at comparing the expression of prolactin receptors in normal, benign, and malignant breast tissue. MATERIAL/METHODS: The expression of prolactin receptors was studied in paraffin wax embedded sections of 102 breast biopsies (93 female and nine male), using the monoclonal antibody B6.2, and the avidin-biotin immunoperoxidase technique. Six biopsies were normal, 34 had benign lesions, and 62 were malignant. RESULTS: In normal cases, prolactin receptor positivity was seen only on the luminal borders of the epithelial cells lining ducts and acini. In most benign lesions, variable degrees of luminal and cytoplasmic staining were seen. Cells showing apocrine metaplasia and florid regular ductal epithelial hyperplasia were mostly negative. In malignant cases, the staining pattern was mostly cytoplasmic and heterogeneous. Forty one of the 59 carcinomas in women showed a degree of positivity involving 10-100% of the tumour cells. A significant direct correlation was found between prolactin receptor and oestrogen receptor staining when only cases that scored more than 100/300 for the latter receptor, using the H scoring system, were considered (p = 0.0207). No correlation was found between prolactin receptors and progesterone receptors, patient's age, tumour size, tumour grade, or axillary lymph node status. CONCLUSIONS: Prolactin receptors seem to be expressed at different cellular sites in normal, benign, and malignant breast epithelial cells. The receptor is expressed in more than two thirds of female breast carcinomas, suggesting that it may play a role in the pathogenesis of the disease. The positivity is correlated with moderate and strong staining for oestrogen receptors in tissue sections, but not with other prognostic factors. PMID- 11729218 TI - Prominent dyserythropoiesis in four cases of haemophagocytic lymphohistiocytosis. AB - Haemophagocytic lymphohistiocytosis (HLH) is a disease characterised by peripheral blood pancytopenia secondary to haemophagocytosis of formed blood cells by activated histiocytes. The demonstration of haemophagocytosis may be difficult and the diagnosis may require repeated tissue sampling (including bone marrow, cerebrospinal fluid, lymph nodes, spleen, and liver) and the demonstration of associated clinical or laboratory features. This report describes pronounced dyserythropoiesis in the bone marrow aspirates in four patients with HLH, including familial and secondary cases. In three patients, bone marrow haemophagocytosis was inconspicuous or absent, and the prominent dyserythropoiesis may have suggested an alternative diagnosis. The dyserythropoiesis observed should be added to the constellation of clinical and laboratory features associated with HLH. PMID- 11729219 TI - Lymphoepithelioma-like carcinoma of the vagina. AB - This report describes a lymphoepithelioma-like carcinoma of the vagina. Although such tumours are well described in the cervix this is only the second report of such a neoplasm at this site. Histology showed a well circumscribed lesion composed of syncytial sheets of epithelioid tumour cells with an intense inflammatory infiltrate, largely consisting of T lymphoid cells. In situ hybridisation and immunohistochemistry for Epstein-Barr virus were negative. A review of the literature reveals that such neoplasms appear to be extremely rare within the female genital tract outside of the cervix. PMID- 11729220 TI - Primitive small round cell tumour of the adrenal gland presenting with fever of unknown origin and t(12;22)(q13;q12) cytogenetic finding. AB - This report describes a left adrenal tumour in a 16 year old Chinese girl who presented with fever of unknown origin. The histological and ultrastructural features of the adrenal tumour were those of a primitive small round cell tumour with neuroendocrine differentiation. Cytogenetic analysis of cultured tumour cells showed a reciprocal translocation t(12;22)(q13;q12). This is the first example of such a tumour being reported in the adrenal gland. The adrenal tumour was also the cause of the fever, which subsided after the removal of the tumour. PMID- 11729221 TI - Increasing recognition of haemoglobin Le Lamentin. PMID- 11729223 TI - Structure, promoter analysis, and chromosomal localization of the murine H(+)/K(+)-ATPase alpha 2 subunit gene. AB - The H(+)/K(+)-ATPase alpha2 subunit (HK alpha 2) of distal colon and renal collecting ducts plays a critical role in potassium and acid-base homeostasis. The isolation and complete sequence of the murine HK alpha 2 gene are reported. The HK alpha 2 gene contains 23 exons and spans 23.5 kb of genomic DNA. The exon/intron organization is comparable to that of the human ATP1AL1 gene. Primer extension and 5'-rapid amplification of cDNA ends of distal colon RNA were used to map the transcription initiation site. Fluorescence in situ hybridization analysis localized the HK alpha 2 gene to murine chromosome 14C3. Sequence analysis of 7.2 kb of the 5'-flanking region revealed numerous consensus sites for transcription factors, including two potential glucocorticoid response elements. Transient transfection of promoter-luciferase constructs demonstrated strong basal HK alpha 2 promoter activity in renal collecting duct cells but not in fibroblasts or in a medullary thick ascending limb of Henle's loop cell line. Deletion analysis revealed that the proximal 0.2 kb of the promoter was sufficient to confer activity in collecting duct cells. These data should prove important in elucidation of the mechanisms controlling the differential, tissue specific expression of the HK alpha 2 gene. PMID- 11729222 TI - Hepatocyte growth factor alters renal epithelial cell susceptibility to uropathogenic Escherichia coli. AB - The urinary tract is frequently the source of Escherichia coli bacteremia. Bacteria from the urinary tract must cross an epithelial layer to enter the bloodstream. Hepatocyte growth factor (HGF) alters the polarity of Madin-Darby canine kidney (MDCK) epithelial cells. The role of cell polarity in determining renal epithelial resistance to Escherichia coli invasion is not well known. A model of polarized and HGF-treated MDCK epithelial cells grown on filters was used to study the role of epithelial cell polarity during the interaction of nonvirulent (XL1-Blue) and uropathogenic (J96) strains of Escherichia coli with renal epithelium. Basolateral exposure of MDCK cells to J96, but not XL1-Blue, resulted in loss of transepithelial resistance (TER), which was due to epithelial cytotoxicity and not degradation of epithelial junctional proteins by bacterial proteases. Apical exposure to both J96 and XL1-Blue did not alter TER. Pretreatment of polarized MDCK cell monolayers with HGF renders the cells sensitive to loss of TER and cytotoxicity by apical exposure to J96. Analysis by confocal microscopy demonstrated that HGF treatment of MDCK cell monolayers also greatly enhances adherence of J96 to the apical surface of the cell monolayer. These data demonstrate that the basolateral surface of polarized epithelia is more susceptible to J96 cytotoxicity. The data also support the hypothesis that processes that alter epithelial cell polarity increase sensitivity of epithelia to bacterial injury and adherence from the apical compartment. PMID- 11729224 TI - Regulated overexpression of heat shock protein 72 protects Madin-Darby canine kidney cells from the detrimental effects of high urea concentrations. AB - Exposure of renal medullary cells to elevated extracellular NaCl concentrations is associated with increased heat shock protein 72 (HSP72) expression and improved resistance to subsequent exposure to a high urea concentration (600 mM). To establish a causal relationship between HSP72 expression and protection against high urea concentrations, HSP72 was inducibly overexpressed in Madin Darby canine kidney (MDCK) cells, in the absence of hypertonic stress before urea exposure. For this purpose, the human stress-inducible HSP72 gene was cloned downstream from a dexamethasone (DEX)-inducible promoter in the eukaryotic expression vector pLKneo. This construct allowed robust induction of HSP72 by exposure of stably transfected MDCK cells (MDCK-LK72) to 0.1 microM DEX. Increased HSP72 abundance significantly improved survival rates after 24-h exposure of the cells to medium containing 600 mM urea (14 versus 43%). In mock transfected or wild-type cells, DEX had no significant effect on HSP72 abundance or urea resistance. In accordance with those findings, lactate dehydrogenase activity in the supernatant was significantly reduced, compared with appropriate control samples, only in MDCK-LK72 cells overexpressing HSP72. Labeling with annexin V-FITC and propidium iodide, followed by flow cytometry, revealed that overexpression of HSP72 was associated with a reduction in the number of apoptotic-lysed cells, a concomitant retardation of apoptosis, and an increase in the number of viable cells. These data support the view that HSP72, which is very abundant in the renal inner medulla, is an important component of the defense mechanism of medullary cells against extreme concentrations of urea. PMID- 11729225 TI - Blood pressure-independent additive effects of pharmacologic blockade of the renin-angiotensin and endothelin systems on progression in a low-renin model of renal damage. AB - Pharmacologic blockade of the renin and endothelin (ET) systems is an established strategy to interfere with progression of renal failure. In the Heyman nephritis model, additive benefits of decreases in BP with the combination of angiotensin converting enzyme inhibitors (ACE-i) and ET(A) receptor antagonists (ET-RA) were demonstrated. To further investigate these findings and to exclude confounding effects of BP decreases, this issue was reassessed in a low-renin model of subtotal kidney resection. Subtotally nephrectomized (SNX) and sham-operated rats were left untreated or received an ACE-i, an angiotensin II subtype 1 receptor antagonist (AT1-RA), an ET-RA, or combinations thereof (ACE-i plus ET-RA or AT1 RA plus ET-RA). The parameters studied were the glomerulosclerosis index (GSI), tubulointerstitial index, vascular damage index, glomerular geometry, and albumin excretion. After 12 wk, BP values were comparable. Urinary albumin excretion rates were significantly higher for untreated SNX rats (24.3 +/- 31.3 mg/24 h), compared with untreated sham-operated rats (0.71 +/- 0.40 mg/24 h). Rates were significantly lower for all treated, compared with untreated, SNX groups. GSI values were significantly higher for untreated SNX rats than for untreated sham operated rats. ACE-i caused significantly lower GSI in SNX rats (0.46 +/- 0.06), compared with AT1-RA (0.60 +/- 0.10) or ET-RA (0.65 +/- 0.10). GSI values were significantly decreased further with ACE-i plus ET-RA (0.29 +/- 0.09) or AT1-RA plus ET-RA (23 +/- 0.05) treatment. Changes in the tubulointerstitial index and vascular damage index proceeded in parallel. The results document BP-independent effects of the ACE-i and AT1-RA on the GSI and urinary albumin excretion and an effect of the ET-RA on the GSI. The contrasting results suggest different pathogenetic pathways for glomerulosclerosis and albuminuria. The combination of treatments provided superior effects on the GSI and tubulointerstitial index but not on urinary albumin excretion. PMID- 11729226 TI - Vitamin E alleviates renal injury, but not hypertension, during chronic nitric oxide synthase inhibition in rats. AB - Chronic nitric oxide (NO) synthase inhibition in rats causes hypertension, renal vascular injury, and proteinuria. NO deficiency increases superoxide (O(2)(-)) activity, but the effects of antioxidant treatment on renal injury have not been studied in this model. Exposure of rats to N omega-nitro-L-arginine (L-NNA) for 4 d markedly decreased NO-dependent relaxation in aortic rings and increased glomerular and renal interstitial monocyte influx, but renal O(2)(-) activity was not increased. After 7 d, BP and proteinuria were significantly increased. After 21 d of L-NNA treatment, rats displayed severe hypertension, decreased GFR, marked proteinuria, glomerular ischemia, renal vascular and tubulointerstitial injury, and complete loss of NO-dependent relaxation. Renal O(2)(-) activity was markedly increased [lucigenin-enhanced chemiluminescence (LEC), 279 +/- 71 versus 50 +/- 7 counts/10 mg, P < 0.01; electron paramagnetic resonance spectroscopy, 0.57 +/- 0.05 versus 0.34 +/- 0.04 U/10 mg, P < 0.05]. Apocynin, a specific inhibitor of NADPH oxidase, and diphenyleneiodonium, an inhibitor of flavin containing enzymes, completely inhibited LEC signals in vitro, whereas allopurinol had no effect, indicating that NAD(P)H oxidase plays a major role in superoxide production in the kidney. Endothelial function remained impaired during cotreatment with alpha-tocopherol and there was no effect on hypertension or tubulointerstitial injury, but glomerular ischemia, decreases in GFR, and renal vascular injury were prevented and proteinuria was ameliorated. Renal LEC signals were intermediate between control and L-NNA-alone values (181 +/- 84 counts/10 mg). Chronic NO synthase inhibition in rats results in marked increases in renal cortical O(2)(-) activity, mediated by flavin-dependent oxidases. The absence of early increases in renal O(2)(-) activity, in the presence of endothelial dysfunction and macrophage influx, indicates that increased renal O(2)(-) activity is neither attributable to NO deficiency per se nor solely related to macrophage influx. The improvement of glomerular function and amelioration of renal vasculitis and proteinuria with vitamin E cotreatment indicate that oxidants are involved in the pathogenesis of renal injury in this model. However, markedly impaired endothelial function and unabated hypertension persist with vitamin E treatment and seem to be directly attributable to NO deficiency. PMID- 11729227 TI - Alteration of the soluble guanylate cyclase system in the vascular wall of lead induced hypertension in rats. AB - Low-level lead exposure is a known cause of hypertension that has been associated with increased reactive oxygen species activity and endothelial-dependent vasorelaxation impairment. The effect of lead exposure on the vascular nitric oxide (NO)/cyclic guanocine monophosphate (cGMP) system was analyzed. Wistar rats were exposed to 5 ppm lead acetate in the drinking water during 30 d. Mean arterial BP increased significantly in the lead-treated rats. Relaxation to both acetylcholine and sodium nitroprusside (SNP) was reduced in lead-treated rats; however, the vascular wall of lead-administered rats showed an increased expression of endothelial NO synthase. The expression of both subunits (alpha(1) and beta(1)) of soluble guanylate cyclase (sGC) and the cGMP accumulated in the vascular wall were decreased in lead-treated rats. Cotreatment of lead with vitamin C (3 mmol/L) prevented the increase on mean arterial BP, improved the relaxation to both acetylcholine and sodium nitroprusside, and restored the normal expression of endothelial NO synthase and sGC proteins in the vascular wall. In conclusion, lead exposure altered both the endothelium-dependent and independent relaxing response and induced a reduced expression of sGC in the vascular wall. These effects were abrogated with the antioxidant vitamin C, which suggests the involvement of reactive oxygen species in the regulation of the NO/cGMP relaxing system in the vascular wall of lead-treated rats. PMID- 11729228 TI - Cloning and characterization of a novel subunit of protein serine/threonine phosphatase 4 from mesangial cells. AB - Mesangial cells play an important role in maintaining glomeruli structure and function and in the pathogenesis of glomerular diseases. With a novel approach using a rapid large-scale DNA sequencing strategy and computerized data processing, a new human gene, PP4(Rmeg) was cloned. The full-length cDNA clone of human PP4(Rmeg) coded for a novel 950-amino acid protein, which was similar to a subunit of protein serine/threonine phosphatase 4 (PP4). Recombinant PP4(Rmeg) produced in COS-7 cells bound to the catalytic subunit of PP4. PP4(Rmeg) is therefore structurally and functionally related to the recently reported regulatory subunit of PP4, PP4(R1). Amino acid sequence analysis of rat PP4(Rmeg) homologue revealed that the sequences were well conserved between human and rat (86.3% identity). Northern blot analyses of human tissues and cultured cells demonstrated that the regulatory subunits were expressed abundantly in human cultured mesangial cells, although their expression was relatively ubiquitous. In situ hybridization studies in normal human renal tissues confirmed their expression in glomeruli in vivo. The expression was upregulated in glomeruli of anti-Thy1 glomerulonephritis rats before mesangial proliferation. These data demonstrate that PP4(Rmeg) is a novel regulatory subunit of PP4, which is expressed ubiquitously but abundantly in mesangial cells. Its pathophysiologic role in mesangial cells and glomerulus remains unknown. As PP4 is an essential protein for nucleation, growth, and stabilization of microtubules at centrosomes/spindle pole bodies during cell division, PP4(Rmeg) may play a role in regulation of mitosis in mesangial cells. PMID- 11729229 TI - Regulation of vascular proteoglycan synthesis by angiotensin II type 1 and type 2 receptors. AB - Recent studies have shown that proteoglycans play an important role in the development of vascular disease and renal failure. In this study, the effects of angiotensin II (AngII) type 1 (AT1) and type 2 (AT2) receptor stimulation on glycosaminoglycan and proteoglycan core protein synthesis in vascular smooth muscle cells (VSMC) were examined. Treatment of AT1 receptor-expressing VSMC with AngII resulted in a dose-dependent and time-dependent increase (2- to 4-fold) in (3)H-glucosamine/(35)S-sulfate incorporation, which was abolished by pretreatment with the AT1 receptor antagonist, losartan. The effects of AngII were inhibited by the epidermal growth factor receptor inhibitor, AG1478, and the mitagen activated protein kinase kinase inhibitor, PD98059, but not the protein kinase C inhibitors, chelerythrine and staurosporine. AngII treatment also resulted in significant increases in the mRNA of the core proteins, versican, biglycan, and perlecan. The effects of AT2 receptor stimulation were examined by retroviral transfection of VSMC with the AT2 receptor. Stimulation of the AT2 receptor in these VSMC-AT2 cells resulted in a significant (1.3-fold) increase in proteoglycan synthesis, which was abolished by the AT2 receptor antagonist, PD123319, and attenuated by pretreatment with pertussis toxin. These results implicate both AT1 and AT2 receptors in the regulation of proteoglycan synthesis and suggest the involvement of epidermal growth factor receptor-dependent tyrosine kinase pathways and G alpha i/o-mediated mechanisms in the effects of the two receptors. PMID- 11729230 TI - Nitric oxide increases albumin permeability of isolated rat glomeruli via a phosphorylation-dependent mechanism. AB - Nitric oxide (NO) has been implicated in the induction of proteinuria in acute inflammatory glomerulonephritis and in the increased vascular permeability seen in various other disease conditions. The complicated interactions of NO with other factors in vivo hinder analysis of the mechanisms involved. By use of a recently introduced method for measuring albumin permeability (P(a)) in isolated glomeruli, the question of whether NO has a direct effect on the permeability barrier of glomerular tufts was examined and the potential mechanisms were explored. Exposure of isolated glomeruli to three NO donors, s-nitroso-N-acetyl penicillamine (SNAP), (Z)-1-[-2-(aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium 1,2-diolate (DETA-NONOate), and sodium nitroprusside, all increased the P(a). This action of NO was time- and concentration-dependent and could be mimicked by 8-bromoguanosine 3', 5'-cyclic monophosphate. Western blot analysis of the proteins from NO donor-treated glomeruli revealed an increase of phosphotyrosine levels of proteins of molecular mass about 120 and 70 kD. The demonstration that pretreatment of glomeruli with the tyrosine kinase inhibitor, genistein, could largely prevent the effect of SNAP and DETA-NONOate confirmed the crucial role of tyrosine phosphorylation in the NO-induced increase of P(a). Furthermore, the tyrosine phosphatase inhibitor, phenylarsine oxide (PAO), could mimic the action of NO on P(a). NO-enhanced tyrosine phosphorylation was further confirmed by immunofluorescence staining, where positive cells in SNAP- and PAO-treated glomeruli were much more frequent than that in controls. By use of dual-label staining in combination with podocyte specific marker, nephrin, it was observed that most of the phosphorylated positive cells corresponded to podocytes. These results suggest that NO impairs the glomerular permeability barrier through a tyrosine phosphorylation-dependent mechanism. PMID- 11729231 TI - Bone marrow is a reservoir of repopulating mesangial cells during glomerular remodeling. AB - The renal glomerulus, whose cellular components are developmentally derived from the mesenchyme, plays a pivotal role in filtratating plasma. Irretrievable changes of glomerular components are responsible for the initiation and progression of impaired renal function. Recently, it has been shown that functional stem cells exist in the bone marrow of adult bodies and that they can reconstitute damaged tissues of the mesenchymal origin. To examine whether the bone marrow provides stem cells to damaged glomeruli, transgenic rats carrying enhanced green fluorescence protein (EGFP rat) were established in a systemic and constitutive manner. After transplanting the bone marrow of EGFP rats into wild type rats, the progeny of the transplanted marrow cells were tracked with a tag of EGFP. Recruitment of bone marrow-derived cells into glomeruli was dramatically facilitated in response to mesangiolysis evoked in anti-Thy1 antibody-mediated glomerulonephritis. In the restored glomeruli, 11% to 12% of glomerular cells were derived from the transplanted bone marrow. The number of bone marrow-derived CD45(+) cells transiently increased during the disease process, and CD45-negative cells constantly accounted for more than half of the bone marrow-derived population in glomeruli. Bone marrow-derived Thy1(+) cells kept increasing in number until the remodeling ceased and finally made up 7% to 8% of glomerular cells. Laser scanning microscopy displayed that the bone marrow-derived Thy1(+) cells provide structural support for glomerular capillaries, which indicates that they are mesangial cells. Although CD45(-)Thy1(-) bone marrow-derived cells exist during the remodeling of glomeruli, none of them expressed endothelial markers such as Factor VIII and RECA1 as long as they were tested. The results indicate that the bone marrow can give rise to mesangial cells in vivo. PMID- 11729232 TI - Human glomerulonephritis accompanied by active cellular infiltrates shows effector T cells in urine. AB - Leukocyturia is associated with postinfectious glomerulonephritis (GN), interstitial nephritis, and renal allograft rejection. In addition, prominent infiltration of T cells and macrophages is commonly observed in the renal tissues of patients with GN, accompanied by cellular crescent formation and/or interstitial cell infiltration. Because these infiltrating T cells were thought to participate in the development of the diseases and to appear in the urinary space while functioning as effector cells in the renal inflammatory lesion, the study focused on the characterization of T cells that appeared in urine. Freshly voided urine cells were analyzed by flow cytometry to determine their phenotype and by reverse transcriptase-PCR to detect cytokine mRNA. In urine from patients with different forms of GN, including IgA nephropathy, Henoch-Schonlein purpura nephritis, and anti-neutrophil cytoplasmic antibody-associated GN, T cells appeared together with macrophages. The urine T cells were mainly CD45RA(-), CD45RO(+), and CD62L (L-selectin)(-), which are the phenotypic features of effector T cells. In agreement with this finding, T cells infiltrating glomeruli, crescents, and tubulointerstitial lesions were also effector type. Moreover, these urine cells expressed mRNA of the T helper lymphocyte 1 cytokines, interleukin-2, and/or interferon-gamma. These findings suggest that the appearance of effector T cells in urine may reflect the cellular immune reaction that occurs in the kidneys of patients with GN accompanied by active cell infiltration. PMID- 11729233 TI - Microcyst formation and HIV-1 gene expression occur in multiple nephron segments in HIV-associated nephropathy. AB - Tubular microcyst formation is a prominent histopathologic feature of HIV associated nephropathy (HIVAN), but its pathogenesis is unknown. HIV-1 has recently been shown to infect renal tubular epithelial cells in patients with HIVAN. In addition, HIV-1 gene expression in renal epithelial cells has been shown to cause a renal disease that is identical to HIVAN in HIV-1 transgenic mice. In these studies, immunohistochemistry for tubular segment-specific markers and mRNA in situ hybridization for HIV-1 was used to determine which tubular segments develop microcysts and which segments express HIV-1 in the kidneys of transgenic mice and patients with HIVAN. It was found that microcysts involve multiple nephron segments in both patients with HIVAN and HIV-1 transgenic mice. Furthermore, HIV-1 infection in HIVAN and HIV-1 transgene expression also occurs in multiple segments of the nephron. These data support a direct role for HIV-1 infection of renal epithelial cells in the pathogenesis of microcyst formation in patients with HIVAN. PMID- 11729234 TI - Overexpression of brain natriuretic peptide in mice ameliorates immune-mediated renal injury. AB - One of major causes of end-stage renal disease is glomerulonephritis, the treatment of which remains difficult clinically. It has already been shown that transgenic mice that overexpress brain natriuretic peptide (BNP), with a potent vasorelaxing and natriuretic property, have ameliorated glomerular injury after subtotal nephrectomy. However, the role of natriuretic peptides in immune mediated renal injury still remains unknown. Therefore, the effects of chronic excess of BNP on anti-glomerular basement membrane nephritis induced in BNP transgenic mice (BNP-Tg) were investigated and the mechanisms how natriuretic peptides act on mesangial cells in vitro were explored. After induction of nephritis, severe albuminuria (approximately 21-fold above baseline), tissue damage, including mesangial expansion and cell proliferation, and functional deterioration developed in nontransgenic littermates. In contrast, BNP-Tg exhibited much milder albuminuria (approximately fourfold above baseline), observed only at the initial phase, and with markedly ameliorated histologic and functional changes. Up-regulation of transforming growth factor-beta (TGF-beta) and monocyte chemoattractant protein-1 (MCP-1), as well as increased phosphorylation of extracellular signal-regulated kinase (ERK), were also significantly inhibited in the kidney of BNP-Tg. In cultured mesangial cells, natriuretic peptides counteracted the effects of angiotensin II with regard to ERK phosphorylation and fibrotic action. Because angiotensin II has been shown to play a pivotal role in the progression of nephritis through induction of TGF-beta and MCP-1 that may be ERK-dependent, the protective effects of BNP are likely to be exerted, at least partly, by antagonizing the renin-angiotensin system locally. The present study opens a possibility of a novel therapeutic potential of natriuretic peptides for treating immune-mediated renal injury. PMID- 11729235 TI - Primary gene structure and expression studies of rodent paracellin-1. AB - The novel member of the claudin multigene family, paracellin-1/claudin-16, encoded by the gene PCLN1, is a renal tight junction protein that is involved in the paracellular transport of magnesium and calcium in the thick ascending limb of Henle's loop. Mutations in human PCLN1 are associated with familial hypomagnesemia with hypercalciuria and nephrocalcinosis, an autosomal recessive disease that is characterized by severe renal magnesium and calcium loss. The complete coding sequences of mouse and rat Pcln1 and the murine genomic structure are here presented. Full-length cDNAs are 939 and 1514 bp in length in mouse and rat, respectively, encoding a putative open-reading frame of 235 amino acids in both species with 99% identity. Exon-intron analysis of the human and mouse genes revealed a 100% homology of coding exon lengths and splice-site loci. By radiation hybrid mapping, the murine Pcln1 gene was assigned directly to marker D16Mit133 on mouse chromosome 16 (syntenic to a locus on human chromosome 3q27, which harbors the human PCLN1 gene). Mouse multiple-tissue Northern blot showed Pcln1 expression exclusively in the kidney. The expression profile along the nephron was analyzed by reverse transcriptase-PCR on microdissected nephron segments and immunohistochemistry of rat kidney. Paracellin-1 expression was restricted to distal tubular segments including the thick ascending limb of Henle's loop, the distal tubule, and the collecting duct. The identification and characterization of the rodent Pcln1 genes provide the basis for further studies of paracellin-1 function in suitable animal models. PMID- 11729236 TI - Temporally compartmentalized expression of ephrin-B2 during renal glomerular development. AB - Glomerular development proceeds through the spatially ordered and sequential recruitment, proliferation, assembly, and differentiation of endothelial, mesangial, and epithelial progenitors. The molecular determinants of cell-cell recognition and targeting in this process have yet to be defined. The Eph/ephrin family of membrane receptors and counter-receptors are critical participants of developmental vascular assembly in extrarenal sites. Renal expression patterns of ephrin-B2 and EphB4 were investigated using mice expressing beta-galactosidase under control of ephrin-B2 or EphB4 promoters. The earliest glomerular expression of ephrin-B2 was identified in a subset of differentiating comma-stage glomerular epithelial cells (podocyte progenitors) adjacent to the vascular cleft where endothelial progenitors are subsequently recruited. Epithelial ephrin-B2 expression was accompanied by expression in endothelial and mesangial cells as capillary assembly progressed. At or near completion of glomerular maturation, epithelial ephrin-B2 expression was extinguished, with persistence in glomerular endothelial cells. Throughout development, one of several ephrin-B2 receptors, EphB4, was persistently and exclusively expressed in endothelial cells of venous structures. The findings show sequential ephrin-B2 expression across glomerular lineages, first in a distinct subset of podocyte progenitors and subsequently in endothelial cells of the developing glomerulus. Given targeting functions for Eph/ephrin family proteins, the findings suggest that ephrin-B2 expression marks podocyte progenitors at the site of vascular cleft formation, where expression may establish an "address" to which endothelial and mesangial progenitors are recruited. Thus, the present results suggest that ephrin-B2 and EphB interactions play an important role in glomerular microvascular assembly. PMID- 11729237 TI - Manganese superoxide dismutase attenuates Cisplatin-induced renal injury: importance of superoxide. AB - Cisplatin is a potent chemotherapeutic agent that is used to treat many human malignancies. Unfortunately, in addition to side effects such as ototoxicity, anaphylaxis, and bone marrow suppression, a significant percentage of patients receiving cisplatin develop severe nephrotoxicity. Mitochondrial dysfunction that is mediated via the generation of reactive oxygen species has been implicated in the pathogenesis of cisplatin-induced renal injury. To address the mechanism, it was hypothesized that overexpression of antioxidant enzymes, such as mitochondria localized manganese superoxide dismutase (MnSOD) or mitochondria-targeted catalase (mito-Cat), would be cytoprotective in cisplatin-induced cell injury. To this end, human MnSOD or a mito-Cat vector were stably transfected into human embryonic kidney 293 cells. Cells that overexpressed MnSOD exhibited significantly less cell rounding and detachment compared with both mito-Cat and vector controls after exposure to 20 microM cisplatin. Cell injury as assessed by DNA fragmentation and annexin V binding assays was significantly decreased in the cells that overexpressed MnSOD compared with vector alone and mito-Cat. In addition, elevated levels of MnSOD were strongly associated with increased clonogenic potential after cisplatin challenge. Thus, overexpression of MnSOD, and not catalase, protects against cisplatin-induced renal epithelial cell injury. These results demonstrate the importance of reactive oxygen species in the mechanism that underlies cisplatin-induced renal injury and specifically implicate the superoxide radical, and not hydrogen peroxide, as the mediator. PMID- 11729238 TI - Cu/Zn-superoxide dismutase gene attenuates ischemia-reperfusion injury in the rat kidney. AB - Evidence has accumulated for a role of toxic oxygen radicals in the pathogenesis of ischemia-reperfusion injury in the kidney. The aim of this study was to evaluate the hypothesis that reducing postischemic renal injury is possible by delivery of the gene for the antioxidant enzyme superoxide dismutase (SOD). Female Sprague-Dawley rats received intravenous injections of recombinant adenovirus (1 x 10(9) pfu) containing the transgenes for Escherichia coli beta galactosidase (Ad-LacZ, as control) or human Cu/Zn-SOD (Ad-SOD). Three days later, renal ischemia was produced by cross-clamping the left renal vessels for 60 min. The right kidney was removed before reperfusion and processed for the transgene. Renal SOD protein and activity in rats given Ad-SOD was 2.5-fold higher than from the animals receiving Ad-LACZ: Urinary lactate dehydrogenase concentrations were elevated by ischemia-reperfusion in the Ad-LacZ group (1403 +/- 112 U/L), yet values were 50% lower in Ad-SOD-treated rats. Free radical production was elevated by ischemia-reperfusion but was significantly lower in SOD-treated animals. Importantly, on postischemic day 1, glomerular filtration rates were reduced to 0.21 ml/min per 100 g in the Ad-LacZ group, whereas values remained significantly higher (0.39) in the Ad-SOD group. Two weeks after ischemia-reperfusion, inflammation, interstitial fibrosis, tubular atrophy and tissue levels of tumor necrosis factor alpha and interleukin-1 were significantly higher in the Ad-LacZ-treated than in Ad-SOD-treated rats. In conclusion, these results indicate that SOD expression can be increased by delivery of the sod gene to the kidney by intravenous injection and that sod gene transduction minimized ischemia-reperfusion-induced acute renal failure. PMID- 11729239 TI - Angiotensin II activates collagen type I gene in the renal cortex and aorta of transgenic mice through interaction with endothelin and TGF-beta. AB - Hypertension is frequently associated with the development of renal vascular fibrosis. This pathophysiologic process is due to the abnormal formation of extracellular matrix proteins, mainly collagen type I. In previous studies, it has been observed that the pharmacologic blockade of angiotensin II (Ang II) or endothelin (ET) blunted the development of glomerulo- and nephroangiosclerosis in nitric oxide-deficient hypertensive animals by inhibiting collagen I gene activation. The purpose of this study was to investigate whether and how AngII interacts with ET to activate the collagen I gene and whether transforming growth factor-beta (TGF-beta) could be a player in this interaction. Experiments were performed in vivo on transgenic mice harboring the luciferase gene under the control of the collagen I-alpha 2 chain promoter (procol alpha 2[I]). Bolus intravenous administration of AngII or ET produced a rapid, dose-dependent activation of collagen I gene in aorta and renal cortical slices (threefold increase over control at 2 h, P < 0.01). The AngII-induced effect on procol alpha 2(I) was completely inhibited by candesartan (AngII type 1 receptor antagonist) and substantially blunted by bosentan (dual ET receptor antagonist) (P < 0.01), whereas the ET-induced activation of collagen I gene was blocked only by bosentan. In subsequent experiments, TGF-beta (also administered intravenously) produced a rapid increase of procol alpha 2(I) in aorta and renal cortical slices (twofold increase over control at 1 h, P < 0.01) that was completely blocked by decorin (scavenger of the active form of TGF-beta). In addition, decorin attenuated the activation of collagen I gene produced by AngII (P < 0.01). These data indicate that AngII can activate collagen I gene in aorta and renal cortex in vivo by a mechanism(s) requiring participation and/or cooperation of ET and TGF-beta. PMID- 11729240 TI - Albuminuria in mice after injection of antibodies against aminopeptidase A: role of angiotensin II. AB - It has been shown that injection of combinations of anti-aminopeptidase A (APA) monoclonal antibodies (mAb) that inhibit the enzyme activity induces an acute albuminuria in mice. This albuminuria is not dependent on inflammatory cells, complement, or the coagulation system. APA is an important regulator of the renin angiotensin system because it is involved in the degradation of angiotensin II (Ang II). This study examined the potential role of glomerular Ang II in the induction of albuminuria. The relation among renal Ang II, glomerular APAX enzyme activity, and albuminuria was examined first. Injection of the nephritogenic combinations ASD-3/37 and ASD-37/41 in BALB/c mice induced albuminuria, whereas the non-nephritogenic combination ASD-3/41 had no effect. There was no clear relation between the inhibition of glomerular APA activity and albuminuria, yet it was evident that intrarenal Ang II levels were significantly increased in albuminuric mice and not in nonalbuminuric mice. As a next step, anti-APA mAb were administered to angiotensinogen-deficient mice that do not produce Ang II, and kidney morphology and albuminuria were determined. Angiotensinogen-deficient mice also developed albuminuria upon ASD-37/41 administration. Altogether, these findings clearly demonstrate that Ang II is not required for the induction of albuminuria upon injection of enzyme-inhibiting anti-APA mAb. PMID- 11729241 TI - Increased renal angiopoietin-1 expression in folic acid-induced nephrotoxicity in mice. AB - Growth factors affect epithelial regeneration after acute renal injury, but less is known regarding the expression of vascular growth factors in this setting. A mouse model of folic acid (FA)-induced nephrotoxicity was used to study the expression of angiopoietins (Ang), factors that bind the Tie-2 receptor and modulate endothelial growth. Tubular damage was detected 1 d after FA administration; in the next 14 d, most tubules regenerated but patchy atrophy, with interstitial fibrosis, was also observed. Ang-1 immunostaining was detected between cortical tubules and in the vasa rectae of vehicle-treated animals. FA induced nephropathy was associated with the acquisition of Ang-1 immunostaining in renal arterial walls and in a subset of injured cortical tubules that failed to stain with periodic acid-Schiff stain, which indicated that they were distal tubules. Renal Ang-1 protein levels were significantly increased after FA administration, compared with time-matched control values, as assessed by Western blotting. Capillaries between regenerating tubules expressed both Tie-2 and platelet-endothelial cell adhesion molecule. A subset of these endothelia expressed proliferating cell nuclear antigen, whereas capillary proliferation was absent in control samples. Therefore, FA-induced nephropathy is associated with increased Ang-1 protein expression in renal epithelia and arteries. In addition, Tie-2-expressing capillaries near damaged cortical tubules undergo proliferation. Further experiments are required to establish whether these events are functionally related. PMID- 11729242 TI - Cyclosporine A inhibits the adaptive responses to hypertonicity: a potential mechanism of nephrotoxicity. AB - Cell survival in the hypertonic environment of the renal medulla is dependent on the intracellular accumulation of protective organic solutes through the induction of genes whose transcriptional regulation is mediated in part by interaction between osmotic response elements and the transcription nuclear factor of activated T lymphocyte 5. It is shown that cyclosporine A (CsA) prevents the nuclear translocation of the transcription nuclear factor of activated T lymphocyte 5 and inhibits osmotic response element-mediated reporter gene expression. The expression of mRNA for hypertonicity-induced genes (aldose reductase, betaine/gamma-amino-n-butyric acid transporter 1, and heat shock protein 70) is also decreased in the medulla of CsA-treated rats. CsA inhibits the increase of betaine/gamma-amino-n-butyric acid transporter 1 and heat shock protein 70 mRNA in osmotically stressed MDCK cells, blocks cell proliferation under isotonic conditions, and augments hypertonicity-induced apoptosis. Histologic examination of the kidneys of CsA-treated rats shows a marked increase in apoptosis in the renal medulla where hypertonicity normally prevails. The data are consistent with calcineurin-mediated induction of hypertonic stress-response genes, and they suggest that CsA nephrotoxicity may in part result from inhibition of the adaptive responses to hypertonicity occurring during the urinary concentrating mechanism. PMID- 11729243 TI - Prevalence, genetics, and clinical features of patients carrying podocin mutations in steroid-resistant nonfamilial focal segmental glomerulosclerosis. AB - Podocin mutations (NPHS2 gene) are responsible for the autosomal recessive form of steroid-resistant nephrotic syndrome. As a result of a screening for these gene alterations in a cohort of Italian patients with nonfamilial nephrotic syndrome and histologic focal segmental glomerulosclerosis (FSGS), nine patients with NPHS2 gene homozygous or composite heterozygous mutations were found. In addition to the previously described defects, two novel mutations at exon 4 were identified (frameshift, L169P); four single nucleotide polymorphisms (SNPs) and one dinucleotide repeat were also identified. On the basis of haplotype analysis, a founder effect was suggested for the 419delG mutation, the most frequently observed in the patients studied. Patients carrying NPHS2 mutations and without a family history of nephrotic syndrome were indistinguishable from those with idiopathic FSGS on the basis of the clinical phenotype. Two of the nine patients had normal renal function at 3 and 10 yr of age, despite the presence of the nephrotic syndrome. The other seven had reached end-stage renal failure at a mean age of 9.6 yr (range, 4 to 17 yr) and had received renal allografts. In those presenting with end-stage renal failure, the clinical and laboratory features both before and after transplantation were similar, including the age at onset, the amount of proteinuria, and the absence of any response to steroids and other immunosuppressants. Finally, two children presented recurrence of mild proteinuria after transplantation, which promptly remitted after plasmapheresis combined with cyclophosphamide. These data demonstrate that podocin mutations in nonfamilial cases of steroid-resistant nephrotic syndrome are frequent and may be due in one case to a founder effect. The pretransplantation and posttransplantation outcomes in the group of patients with mutations of the podocin gene are similar to classical idiopathic FSGS, including the possibility of recurrence of proteinuria that is mild and responsive to plasmapheresis. These observations support a role of molecular screening of the podocin gene in patients with nephrotic syndrome before immunosuppressive treatment is started. PMID- 11729244 TI - Oxidative stress and endothelial function in chronic renal failure. AB - This study aimed to investigate the relationship between oxidative stress and endothelium-dependent vasodilation in patients with chronic renal failure (CRF). Thirty-seven patients with CRF underwent evaluation of endothelium-dependent vasodilation and endothelium-independent vasodilation by means of forearm blood flow measurements with venous occlusion plethysmography during local intra arterial infusions of methacholine (evaluating endothelium-dependent vasodilation) and sodium nitroprusside (evaluating endothelium-independent vasodilation). Lag phase of lipoprotein fraction to oxidation, total antioxidative activity, diene conjugates, thiobarbituric acid reactive substances, lipid hydroperoxide, reduced glutathione (GSH), oxidized GSH (GSSG), and the GSH redox ratio (GSSG/GSH) were all measured as markers of oxidative stress. Two groups of healthy subjects (61 and 37 subjects, respectively) were used as controls. In one group, oxidative stress markers were measured, whereas endothelium-dependent vasodilation and endothelium-independent vasodilation were assessed in the other group. Compared with controls, the patients with renal insufficiency had an impaired endothelium-dependent vasodilation, a shorter lag phase of lipoprotein fraction, and higher levels of diene conjugates, lipid hydroperoxide, and GSSG levels. The GSSG/GSH ratio was lower in patients with CRF. Endothelium-dependent vasodilation was positively correlated with total antioxidative activity (r = 0.41, P = 0.016), GSH (r = 0.44, P < 0.0098), and lag phase of LDL (r = 0.35, P = 0.036) and negatively correlated with GSSG (r = 0.40, P < 0.018), GSSG/GSH (r = -0.47, P = 0.0057), and diene conjugates (r = 0.53 P < 0.0015) in patients with CRF. These results show that an impaired endothelium vasodilation function and oxidative stress are related to each other in patients with CRF. PMID- 11729245 TI - Forecast of the number of patients with end-stage renal disease in the United States to the year 2010. AB - As the United States end-stage renal disease (ESRD) program enters the new millennium, the continued growth of the ESRD population poses a challenge for policy makers, health care providers, and financial planners. To assist in future planning for the ESRD program, the growth of patient numbers and Medicare costs was forecasted to the year 2010 by modeling of historical data from 1982 through 1997. A stepwise autoregressive method and exponential smoothing models were used. The forecasting models for ESRD patient numbers demonstrated mean errors of -0.03 to 1.03%, relative to the observed values. The model for Medicare payments demonstrated -0.12% mean error. The R(2) values for the forecasting models ranged from 99.09 to 99.98%. On the basis of trends in patient numbers, this forecast projects average annual growth of the ESRD populations of approximately 4.1% for new patients, 6.4% for long-term ESRD patients, 7.1% for dialysis patients, 6.1% for patients with functioning transplants, and 8.2% for patients on waiting lists for transplants, as well as 7.7% for Medicare expenditures. The numbers of patients with ESRD in 2010 are forecasted to be 129,200 +/- 7742 (95% confidence limits) new patients, 651,330 +/- 15,874 long-term ESRD patients, 520,240 +/- 25,609 dialysis patients, 178,806 +/- 4349 patients with functioning transplants, and 95,550 +/- 5478 patients on waiting lists. The forecasted Medicare expenditures are projected to increase to $28.3 +/- 1.7 billion by 2010. These projections are subject to many factors that may alter the actual growth, compared with the historical patterns. They do, however, provide a basis for discussing the future growth of the ESRD program and how the ESRD community can meet the challenges ahead. PMID- 11729246 TI - Alterations of left ventricular hypertrophy in and survival of patients receiving hemodialysis: follow-up of an interventional study. AB - Left ventricular (LV) hypertrophy (LVH) is a risk factor for mortality in patients with end-stage renal disease (ESRD). Whether the attenuation of LVH has a positive effect on survival of patients with ESRD has not been documented. The aim of this study was to determine the effect of parallel treatment of hypertension and anemia on LV mass (LVM) and to determine the effect of LVM changes on survival. A cohort of 153 patients receiving hemodialysis was studied. The duration of follow-up was 54 +/- 37 mo. All patients had echocardiographic determination of LV dimensions and LVM at baseline and regular intervals until the end of the follow-up period. During the study, BP decreased from (mean +/- SD) 169.4 +/- 29.7/90.2 +/- 15.6 to 146.7 +/- 29/78 +/- 14.1 mmHg (P < 0.001), and hemoglobin increased from 8.65 +/- 1.65 to 10.5 +/- 1.45 g/dl (P < 0.001). The LV end-diastolic diameter and mean wall thickness decreased from 56.6 +/- 6.5 to 54.8 +/- 6.5 mm (P < 0.001), and from 10.4 +/- 1.6 to 10.2 +/- 1.6 mm (P < 0.05), respectively. The LVM decreased from 290 +/- 80 to 264 +/- 86 g (P < 0.01). Fifty-eight deaths occurred, 38 attributed to cardiovascular (CV) disease and 20 attributed to non-CV causes. According to Cox analyses after adjustment for age, gender, diabetes, history of CV disease, and all nonspecific CV risk factors, LVM regression positively affected the survival. The hazard risk ratio associated with a 10% LVM decrease was 0.78 (95% confidence interval, 0.63 to 0.92) for all-causes mortality and 0.72 (95% confidence interval, 0.51 to 0.90) for mortality due to CV disease. These results show that a partial LVH regression in patients with ESRD had a favorable and independent effect on patients' all cause and CV survival. PMID- 11729247 TI - Prognostic impact of the indexation of left ventricular mass in patients undergoing dialysis. AB - Left ventricular hypertrophy (LVH) is exceedingly frequent in patients undergoing dialysis. Cardiac mass is proportional to body size, but the influence of various indexing methods has not been studied in patients with end-stage renal disease. The issue is important because malnutrition and volume expansion would both tend to distort the estimate of LV mass (LVM) in these patients. In a cohort of 254 patients, the prognostic impact on all-cause mortality and cardiovascular outcomes of LVH values, calculated according to two established methods of indexing, either body surface area (BSA) or height(2.7), was assessed prospectively. When LVM was analyzed as a categorical variable, the height(2.7) based method identified a larger number of patients with LVH than the corresponding BSA-based method. One hundred and thirty-seven fatal and nonfatal cardiovascular events occurred during the follow-up period. Overall, 90 patients died, 51 of cardiovascular causes. In separate Cox models, both the LVM/height(2.7) and the LVM/BSA index independently predicted total and cardiovascular mortality (P < 0.001). However, the height(2.7)-based method coherently produced a closer-fitting model (P < or = 0.02) than did the BSA-based method. The height(2.7) index was also important for the subcategorization of patients according to the presence of concentric or eccentric LVH because the prognostic value of such subcategorization was apparent only when the height(2.7) based criterion was applied. In conclusion, LVM is a strong and independent predictor of survival and cardiovascular events in patients undergoing dialysis. The indexing of LVM by height(2.7) provides more powerful prediction of mortality and cardiovascular outcomes than the BSA-based method, and the use of this index appears to be appropriate in patients undergoing dialysis. PMID- 11729248 TI - Better preservation of peritoneal morphologic features and defense in rats after long-term exposure to a bicarbonate/lactate-buffered solution. AB - The long-term effects of a standard lactate-buffered dialysis fluid and a new, two-chamber, bicarbonate/lactate-buffered dialysis fluid (with fewer glucose degradation products and a neutral pH) were compared in an in vivo peritoneal exposure model. Rats were given daily injections, via an access port, of 10 ml of standard solution or bicarbonate/lactate-buffered solution for 9 to 10 wk. The omentum, peritoneum, and mesothelial cell layer were screened for morphologic changes. In addition, the bacterial clearing capacity of the peritoneal cells was studied. Significantly more milky spots and blood vessels were observed in the omenta of animals treated with standard solution (P < 0.03 for both parameters). Electron-microscopic analysis demonstrated dramatic changes in the appearance of the vascular endothelial cells of the milky spots and a severely damaged or even absent mesothelium on the peritoneal membrane of the standard solution-treated animals. In contrast, the mesothelium was still present in the bicarbonate/lactate-buffered solution group, although the cells lost microvilli. Both peritoneal dialysis fluids significantly increased the density of mesothelial cells (per square millimeter) on the surface of the liver and the thickness of the submesothelial extracellular matrix of the peritoneum (both P < 0.04 for both fluids versus control). A significantly better ex vivo bacterial clearing capacity was observed with peritoneal cells from the bicarbonate/lactate buffered solution group, compared with the standard solution group (P < 0.05 in both experiments). These results demonstrate that instillation of bicarbonate/lactate-buffered solution into rats for 9 to 10 wk preserves both morphologic and immune parameters much more effectively, compared with standard solution. These findings may be of considerable clinical importance. PMID- 11729249 TI - Glucose and prednisolone alter basic fibroblast growth factor expression in peritoneal mesothelial cells and fibroblasts. AB - The mechanism of peritoneal fibrosis in patients on continuous ambulatory peritoneal dialysis is poorly understood. The production of basic fibroblast growth factor (bFGF) by human peritoneal mesothelial cells cultured in high glucose medium was investigated, and the behavior of peritoneal fibroblasts, as well as the inhibitory effect of prednisolone, was assessed. Reverse transcriptase-PCR and immunocytochemistry showed the expression of glucocorticoid receptors in mesothelial cells. The semiquantitative reverse transcriptase-PCR showed that high glucose medium (4.0%) increased bFGF mRNA by 2.5-fold relative to control medium (0.1% glucose), with 83% suppression of the increase by 1 microM prednisolone. The bFGF protein level in culture supernatant was also increased by 1.5-fold in high glucose medium, with this change showing 45% suppression by 1 microM prednisolone. These effects of prednisolone were prevented by a glucocorticoid receptor antagonist (RU486) in a concentration dependent manner. The proliferation of peritoneal fibroblasts was increased 1.9 fold by the supernatant of mesothelial cells cultured in high glucose medium, with 85% suppression by 1 microM prednisolone and suppression to 16% below basal proliferation by an anti-bFGF neutralizing antibody (10 microg/ml), whereas proliferation showed a concentration-dependent increase on addition of an anti transforming growth factor beta-neutralizing antibody. Recombinant bFGF (50 to 1000 pg/ml) likewise caused a concentration-dependent increase of peritoneal fibroblast proliferation and fibronectin release by these cells was also increased (at 50 to 5000 pg/ml). These results suggest the potential importance of bFGF for initiation of peritoneal fibrosis and the possible efficacy of glucocorticoids for preventing such fibrosis in patients receiving peritoneal dialysis. PMID- 11729250 TI - Association among SF36 quality of life measures and nutrition, hospitalization, and mortality in hemodialysis. AB - Patients on maintenance hemodialysis (MHD) often show substantial reductions in quality of life (QoL). The SF36 (Short Form with 36 questions), a well documented, self-administered QoL scoring system that includes eight independent scales and two main dimensions, has been widely used and validated. In 65 adult outpatients on MHD, the SF36 and its scales and dimensions, scored as a number between 0 and 100, and the nutritional and inflammatory state measured by subjective global assessment, near-infrared (NIR) body fat, body mass index (BMI), and pertinent laboratory values, including hemoglobin, albumin, and C reactive protein were assessed. Twelve-month prospective hospitalization rates and mortality were used as the clinical outcomes. Multivariate (case-mix) adjusted correlation coefficients were statistically significant between SF36 scores and serum albumin and hemoglobin concentrations. There were significant inverse correlations between SF36 scores and the BMI and NIR body fat percentage. Hypoalbuminemic, anemic, and obese patients on MHD had a worse QoL. Prospective hospitalizations correlated significantly with the SF36 total score and its two main dimensions (r between -0.28 and -0.40). The Cox proportional regression relative risk of death for each 10 unit decrease in SF36 was 2.07 (95% CI, 1.08 to 3.98; P = 0.02). Of the eight components and two dimensions of the SF36, the Mental Health dimension and the SF36 total score had the strongest predictive value for mortality. Thus, in patients on MHD the SF36 appears to have significant associations with measures of nutritional status, anemia, and clinical outcomes, including prospective hospitalization and mortality. Even though obesity, unlike undernutrition, is not generally an indicator of poor outcome in MHD, the SF36 may detect obese patients on MHD at higher risk for morbidity and mortality. PMID- 11729251 TI - Flow cytometric crossmatching in primary renal transplant recipients with a negative anti-human globulin enhanced cytotoxicity crossmatch. AB - Flow cytometric crossmatching (FCXM) and panel reactive antibody (PRA) screening techniques are more sensitive than anti-human globulin enhanced cytotoxicity (AHG CDC) techniques at detecting anti-HLA antibodies. The clinical significance of a positive FCXM in primary renal transplant recipients with a negative AHG-CDC crossmatch is unclear. We performed retrospective FCXM and flow cytometric panel reactive antibody (FlowPRA) determinations in primary renal transplant recipients with a negative T cell AHG-CDC crossmatch and a negative B cell CDC crossmatch pretransplant. Eighteen (13%) of 143 patients exhibited a positive retrospective T cell FCXM. Of these patients, six (33%) experienced early graft loss with explant histology, demonstrating antibody-mediated rejection in five of six cases. The 12 patients with positive T cell FCXM who maintained their grafts experienced more adverse events posttransplant, including more early, steroid resistant, and recurrent rejection. Furthermore, in a subgroup of patients undergoing protocol biopsies, those with a positive T cell FCXM exhibited more subclinical rejection. Anti-HLA antibodies were detected by FlowPRA in all 18 patients with a positive T cell FCXM, whereas AHG-CDC PRA detected antibodies in only 8 of 18 patients. Therefore, flow cytometric techniques identify sensitized primary renal transplant recipients undetected by AHG-CDC techniques. In those patients, a positive T cell FCXM is associated with an increased risk of early graft loss due to antibody-mediated rejection and may represent a relative contraindication to transplantation. Moreover, those patients are also at increased risk of severe and recurrent rejection, which may carry implications for long-term graft outcomes. PMID- 11729252 TI - Thymic microchimerism correlates with the outcome of tolerance-inducing protocols for solid organ transplantation. AB - This study found that pretransplant infusion of donor peripheral blood leukocytes, either total leukocytes (peripheral blood leukocytes) or peripheral blood mononuclear cells (PBMC), under appropriate immunomodulating conditions was more effective than donor bone marrow (BM) in prolonging the survival of rats that received kidney grafts. A higher percentage of MHCII(+) cells was found in donor PBMC than in BM cells, and depletion of MHCII(+) cells from donor PBMC abolished their tolerogenic potential. By the analysis of microchimerism in rats infused with donor cells and killed at different time points thereafter, the better tolerogenic potential of leukocyte infusion related to a higher capability of these cells to engraft the recipient thymus. PCR analysis on OX6 immunopurified cells revealed the presence of donor MHCII(+) cells in the thymus of these animals. The role of intrathymic microchimerism was reinforced by findings that thymectomy at the time of transplant prevented tolerance induction by donor leukocytes. Donor DNA was found in the thymus of most long-term graft animals that survived, but in none of those that rejected their grafts. The presence of intrathymic microchimerism correlated with graft survival, and microchimerism in other tissues was irrelevant. PCR analysis of DNA from thymic cell subpopulations revealed the presence of donor MHCII(+) cells in the thymus of long-term surviving animals. Thus, in rats, donor leukocyte infusion is better than donor BM for inducing graft tolerance, defined by long-term graft survival, donor-specific T cell hyporesponsiveness, and reduced interferon gamma production. This effect appears to occur through migration of donor MHCII(+) cells in the host thymus. PMID- 11729253 TI - Suppression of HIV-1 expression by inhibitors of cyclin-dependent kinases promotes differentiation of infected podocytes. AB - The glomerular lesions of HIV-associated nephropathy (HIVAN) are associated with the expression of HIV-1 in podocytes. Infected podocytes proliferate and lose several differentiation markers in vivo and in vitro, which suggests that HIV-1 gene expression induces these changes. Flavopiridol and roscovitine, newly identified inhibitors of cyclin-dependent kinase-9, markedly decrease HIV-1 promoter activity in cell lines of various lineages. In this study, the inhibitors were used to determine whether suppression of HIV-1 transcription in infected podocytes correlated with an inhibition of proliferation and a return to the differentiated phenotype. Dose-response analysis showed that both flavopiridol and roscovitine reversibly suppressed HIV-1 transcription in podocytes in vitro at an IC(50) of 25 nM and 3 microM, respectively. Despite equivalent suppression of HIV-1 transcription, roscovitine was a more effective inhibitor of podocyte proliferation than flavopiridol. Suppression of HIV-1 transcription by flavopiridol or roscovitine was marked by re-expression of the podocyte differentiation markers, synaptopodin and podocalyxin. These results suggest that inhibition of HIV-1 transcription decreases podocyte proliferation and permits the reexpression of differentiation markers. Thus, suppression of HIV 1 transcription by selective cyclin-dependent kinase-9 inhibitors may be a useful therapeutic strategy for the treatment of HIVAN. PMID- 11729254 TI - ACE inhibitors to prevent end-stage renal disease: when to start and why possibly never to stop: a post hoc analysis of the REIN trial results. Ramipril Efficacy in Nephropathy. AB - In this post hoc, secondary analysis of the Ramipril Efficacy In Nephropathy (REIN) trial, an angiotensin-converting enzyme (ACE) inhibition risk/benefit profile was assessed in 322 patients with nondiabetic, proteinuric chronic nephropathies and different degrees of renal insufficiency. The rate of GFR decline (Delta GFR) and the incidence of end-stage renal disease (ESRD) during ramipril or non-ACE inhibitor treatment were compared within three tertiles of basal GFR. Delta GFR was comparable in the three tertiles, whereas the incidence of ESRD was higher in the lowest tertile than in the middle and highest tertiles. Ramipril decreased Delta GFR by 22%, 22%, and 35% and the incidence of ESRD by 33% (P < 0.05), 37%, and 100% (P < 0.01) in the lowest, middle, and highest tertiles, respectively. Delta GFR reduction was predicted by basal systolic (P < 0.0001), diastolic (P = 0.02), and mean (P < 0.001) BP and proteinuria (P < 0.0001) but not by basal GFR (P = 0.12). ESRD risk reduction was predicted by basal proteinuria (P < 0.01) and GFR (P < 0.0001) and was strongly dependent on treatment duration (P < 0.0001). Adverse events were comparable among the three tertiles and within each tertile in the two treatment groups. Thus, disease progression and response to ACE inhibition do not depend on severity of renal insufficiency. The risk of ESRD and the absolute number of events saved by ACE inhibition is highest in patients with the lowest GFR. However, renoprotection is maximized when ACE inhibition is started earlier and when long-lasting treatment may result in GFR stabilization and definitive prevention of ESRD. PMID- 11729255 TI - Lower-extremity peripheral arterial disease among patients with end-stage renal disease. AB - Peripheral arterial occlusive disease (PAOD) accounts for significant morbidity and mortality among end-stage renal disease (ESRD) patients but has not been as extensively studied as other kinds of atherosclerotic disease in this population. The current epidemiology and management of PAOD in ESRD patients is here reviewed and target areas for future research are identified. The prevalence of PAOD appears to be much higher among ESRD patients than in the general population. Risk factors for disease among ESRD patients are not well understood but probably include both conventional and dialysis or uremia-associated risk factors. Standard diagnostic techniques used to identify PAOD in the general population may not be as helpful in ESRD patients because many of these tests are inaccurate in the settings of vascular calcification and small-vessel disease. Despite the fact that this is a common disease in ESRD patients, most of these patients are not screened for PAOD. Interventions that have proven effective in the prevention and treatment of PAOD in the general population, such as smoking cessation, preventive foot care, and exercise, have not been systematically applied to ESRD patients. Furthermore, the optimal management of ischemic ulceration and gangrene in ESRD patients is quite controversial, and better algorithms for the prevention and management of PAOD in ESRD patients are needed. In conclusion, PAOD is common in ESRD patients. Future research should identify risk factors for disease in this population, and efforts should be made to develop strategies for the effective prevention and management of limb ischemia in this population. PMID- 11729257 TI - "Wishing don't make it so"--why we need a randomized clinical trial of high intensity hemodialysis. PMID- 11729258 TI - Sleep apnea in renal patients. PMID- 11729259 TI - A syndrome of renal sodium loss and hyponatremia probably resulting from inappropriate secretion of antidiuretic hormone. 1957. PMID- 11729261 TI - Comparative biology of IncQ and IncQ-like plasmids. AB - Plasmids belonging to Escherichia coli incompatibility group Q are relatively small (approximately 5 to 15 kb) and able to replicate in a remarkably broad range of bacterial hosts. These include gram-positive bacteria such as Brevibacterium and Mycobacterium and gram-negative bacteria such as Agrobacterium, Desulfovibrio, and cyanobacteria. These plasmids are mobilized by several self-transmissible plasmids into an even more diverse range of organisms including yeasts, plants, and animal cells. IncQ plasmids are thus highly promiscuous. Recently, several IncQ-like plasmids have been isolated from bacteria found in environments as diverse as piggery manure and highly acidic commercial mineral biooxidation plants. These IncQ-like plasmids belong to different incompatibility groups but have similar broad-host-range replicons and mobilization properties to the IncQ plasmids. This review covers the ecology, classification, and evolution of IncQ and IncQ-like plasmids. PMID- 11729262 TI - Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. AB - Degradation of plant cell wall polysaccharides is of major importance in the food and feed, beverage, textile, and paper and pulp industries, as well as in several other industrial production processes. Enzymatic degradation of these polymers has received attention for many years and is becoming a more and more attractive alternative to chemical and mechanical processes. Over the past 15 years, much progress has been made in elucidating the structural characteristics of these polysaccharides and in characterizing the enzymes involved in their degradation and the genes of biotechnologically relevant microorganisms encoding these enzymes. The members of the fungal genus Aspergillus are commonly used for the production of polysaccharide-degrading enzymes. This genus produces a wide spectrum of cell wall-degrading enzymes, allowing not only complete degradation of the polysaccharides but also tailored modifications by using specific enzymes purified from these fungi. This review summarizes our current knowledge of the cell wall polysaccharide-degrading enzymes from aspergilli and the genes by which they are encoded. PMID- 11729266 TI - Earthquake recurrence and rupture dynamics of Himalayan Frontal Thrust, India. AB - The Black Mango fault is a structural discontinuity that transforms motion between two segments of the active Himalayan Frontal Thrust (HFT) in northwestern India. The Black Mango fault displays evidence of two large surface rupture earthquakes during the past 650 years, subsequent to 1294 A.D. and 1423 A.D., and possibly another rupture at about 260 A.D. Displacement during the last two earthquakes was at minimum 4.6 meters and 2.4 to 4.0 meters, respectively, and possibly larger for the 260 A.D. event. Abandoned terraces of the adjacent Markanda River record uplift due to slip on the underlying HFT of 4.8 +/- 0.9 millimeters per year or greater since the mid-Holocene. The uplift rate is equivalent to rates of fault slip and crustal shortening of 9.6(-3.5)(+7.0) millimeters per year and 8.4(-3.6)(+7.3) millimeters per year, respectively, when it is assumed that the HFT dips 30 degrees +/- 10 degrees. PMID- 11729267 TI - Formation of recent martian debris flows by melting of near-surface ground ice at high obliquity. AB - The observation of small gullies associated with recent surface runoff on Mars has renewed the question of liquid water stability at the surface of Mars. The gullies could be formed by groundwater seepage from underground aquifers; however, observations of gullies originating from isolated peaks and dune crests question this scenario. We show that these landforms may result from the melting of water ice in the top few meters of the martian subsurface at high obliquity. Our conclusions are based on the analogy between the martian gullies and terrestrial debris flows observed in Greenland and numerical simulations that show that above-freezing temperatures can occur at high obliquities in the near surface of Mars, and that such temperatures are only predicted at latitudes and for slope orientations corresponding to where the gullies have been observed on Mars. PMID- 11729263 TI - Biodegradation of aromatic compounds by Escherichia coli. AB - Although Escherichia coli has long been recognized as the best-understood living organism, little was known about its abilities to use aromatic compounds as sole carbon and energy sources. This review gives an extensive overview of the current knowledge of the catabolism of aromatic compounds by E. coli. After giving a general overview of the aromatic compounds that E. coli strains encounter and mineralize in the different habitats that they colonize, we provide an up-to-date status report on the genes and proteins involved in the catabolism of such compounds, namely, several aromatic acids (phenylacetic acid, 3- and 4 hydroxyphenylacetic acid, phenylpropionic acid, 3-hydroxyphenylpropionic acid, and 3-hydroxycinnamic acid) and amines (phenylethylamine, tyramine, and dopamine). Other enzymatic activities acting on aromatic compounds in E. coli are also reviewed and evaluated. The review also reflects the present impact of genomic research and how the analysis of the whole E. coli genome reveals novel aromatic catabolic functions. Moreover, evolutionary considerations derived from sequence comparisons between the aromatic catabolic clusters of E. coli and homologous clusters from an increasing number of bacteria are also discussed. The recent progress in the understanding of the fundamentals that govern the degradation of aromatic compounds in E. coli makes this bacterium a very useful model system to decipher biochemical, genetic, evolutionary, and ecological aspects of the catabolism of such compounds. In the last part of the review, we discuss strategies and concepts to metabolically engineer E. coli to suit specific needs for biodegradation and biotransformation of aromatics and we provide several examples based on selected studies. Finally, conclusions derived from this review may serve as a lead for future research and applications. PMID- 11729268 TI - Role of diacylglycerol in PKD recruitment to the TGN and protein transport to the plasma membrane. AB - Protein kinase D (PKD) is a cytosolic serine-threonine kinase that binds to the trans-Golgi network (TGN) and regulates the fission of transport carriers specifically destined to the cell surface. PKD was found to bind diacylglycerol (DAG), and this binding was necessary for its recruitment to the TGN. Reducing cellular levels of DAG inhibited PKD recruitment and blocked protein transport from the TGN to the cell surface. Thus, a DAG-dependent, PKD-mediated signaling regulates the formation of transport carriers from the TGN in mammalian cells. PMID- 11729269 TI - Role of apoptosis in Pseudomonas aeruginosa pneumonia. PMID- 11729270 TI - Dealing with the tinder as well as the flint. PMID- 11729265 TI - Actin-based motility of intracellular microbial pathogens. AB - A diverse group of intracellular microorganisms, including Listeria monocytogenes, Shigella spp., Rickettsia spp., and vaccinia virus, utilize actin based motility to move within and spread between mammalian host cells. These organisms have in common a pathogenic life cycle that involves a stage within the cytoplasm of mammalian host cells. Within the cytoplasm of host cells, these organisms activate components of the cellular actin assembly machinery to induce the formation of actin tails on the microbial surface. The assembly of these actin tails provides force that propels the organisms through the cell cytoplasm to the cell periphery or into adjacent cells. Each of these organisms utilizes preexisting mammalian pathways of actin rearrangement to induce its own actin based motility. Particularly remarkable is that while all of these microbes use the same or overlapping pathways, each intercepts the pathway at a different step. In addition, the microbial molecules involved are each distinctly different from the others. Taken together, these observations suggest that each of these microbes separately and convergently evolved a mechanism to utilize the cellular actin assembly machinery. The current understanding of the molecular mechanisms of microbial actin-based motility is the subject of this review. PMID- 11729271 TI - Bioethics. Cloning announcement sparks debate and scientific skepticism. PMID- 11729272 TI - Molecular imaging. Virus infects cell: live and uncut. PMID- 11729264 TI - Transport into and out of the nucleus. AB - A defining characteristic of eukaryotic cells is the possession of a nuclear envelope. Transport of macromolecules between the nuclear and cytoplasmic compartments occurs through nuclear pore complexes that span the double membrane of this envelope. The molecular basis for transport has been revealed only within the last few years. The transport mechanism lacks motors and pumps and instead operates by a process of facilitated diffusion of soluble carrier proteins, in which vectoriality is provided by compartment-specific assembly and disassembly of cargo-carrier complexes. The carriers recognize localization signals on the cargo and can bind to pore proteins. They also bind a small GTPase, Ran, whose GTP-bound form is predominantly nuclear. Ran-GTP dissociates import carriers from their cargo and promotes the assembly of export carriers with cargo. The ongoing discovery of numerous carriers, Ran-independent transport mechanisms, and cofactors highlights the complexity of the nuclear transport process. Multiple regulatory mechanisms are also being identified that control cargo-carrier interactions. Circadian rhythms, cell cycle, transcription, RNA processing, and signal transduction are all regulated at the level of nucleocytoplasmic transport. This review focuses on recent discoveries in the field, with an emphasis on the carriers and cofactors involved in transport and on possible mechanisms for movement through the nuclear pores. PMID- 11729273 TI - Research collaborations. Asian astronomers build closer ties. PMID- 11729274 TI - Ultrafast lasers. Photoelectrons show how quick a flash is. PMID- 11729275 TI - Conservation biology. When is a coho salmon not a coho salmon? PMID- 11729276 TI - Marine conservation. Reserves found to aid fisheries. PMID- 11729277 TI - Genetics. Fragile X's missing partners identified. PMID- 11729279 TI - Anthrax. Can lab sleuths clinch the case? PMID- 11729278 TI - Taking anthrax's genetic fingerprints. PMID- 11729280 TI - A second anthrax genome project. PMID- 11729281 TI - Anthrax. A 'sure killer' yields to medicine. PMID- 11729282 TI - Paleontology. TV dinosaur team treads tricky mammalian terrain. PMID- 11729283 TI - Gamma ray bursts. Cosmic mystery objects start to yield secrets. PMID- 11729285 TI - Gamma ray bursts. Short-lived mysteries. PMID- 11729284 TI - Gamma ray bursts. HETE-2, in business at last. PMID- 11729286 TI - Computing research. Microsoft settles down amongst the dons. PMID- 11729287 TI - Geological Society of America. Life--potential, slow, or long dead. PMID- 11729288 TI - Climate change. Recent reductions in China's greenhouse gas emissions. PMID- 11729289 TI - Cell biology. TAPping into mRNA export. AB - There seem to be numerous pathways for exporting mRNAs from the nucleus to the cytoplasm. But working out which set of export adaptors and receptors transport individual mRNAs has been very difficult. In a Perspective, Moore and Rosbash discuss a new strategy using cell-penetrating peptide inhibitors for unraveling the routes of mRNA export in living cells (Gallouzi and Steitz). PMID- 11729290 TI - Natural product synthesis. The art of total synthesis. AB - Advances in synthetic strategies and analytical technology are allowing ever more natural product molecules to be synthesized. In his Perspective, Marko briefly reviews the history of this field and highlights the synthesis of ciguatoxin by Hirama et al. This marine toxin belongs to the ciguatera family of neurotoxins, which poison some 20,000 people every year. The elegant and highly convergent strategy developed by the authors will be helpful in the development of antibodies to ciguatoxin for the detection of contaminated seafood. PMID- 11729291 TI - Planetary science. Clues to the Martian atmosphere. PMID- 11729292 TI - Paleoclimate. Dating--vive la difference. PMID- 11729293 TI - Signal transduction. A new thread in an intricate web. AB - Understanding how biochemical pathways are connected in the cell is one of the big challenges facing cell biologists. In a Perspective, von Zastrow and Mostov describe new work that identifies a protein called RGS-PX1 as the linchpin that connects signal transduction activated by G protein-coupled receptors with membrane trafficking events. PMID- 11729294 TI - Ecology. Dammed experiments! PMID- 11729295 TI - Immunology. A molecular gut reaction. PMID- 11729296 TI - Network analysis. The structure of the Web. PMID- 11729297 TI - Amersham Biosciences & Science Prize. AMPA receptor dynamics and synaptic plasticity. PMID- 11729298 TI - Amersham Biosciences & Science Prize. 2001 grand prize winner. PMID- 11729300 TI - Caveolae: a once-elusive structure gets some respect. PMID- 11729301 TI - Caveolin-3 helps build muscles. PMID- 11729302 TI - Nuclear receptors and lipid physiology: opening the X-files. AB - Cholesterol, fatty acids, fat-soluble vitamins, and other lipids present in our diets are not only nutritionally important but serve as precursors for ligands that bind to receptors in the nucleus. To become biologically active, these lipids must first be absorbed by the intestine and transformed by metabolic enzymes before they are delivered to their sites of action in the body. Ultimately, the lipids must be eliminated to maintain a normal physiological state. The need to coordinate this entire lipid-based metabolic signaling cascade raises important questions regarding the mechanisms that govern these pathways. Specifically, what is the nature of communication between these bioactive lipids and their receptors, binding proteins, transporters, and metabolizing enzymes that links them physiologically and speaks to a higher level of metabolic control? Some general principles that govern the actions of this class of bioactive lipids and their nuclear receptors are considered here, and the scheme that emerges reveals a complex molecular script at work. PMID- 11729303 TI - Prostaglandins and leukotrienes: advances in eicosanoid biology. AB - Prostaglandins and leukotrienes are potent eicosanoid lipid mediators derived from phospholipase-released arachidonic acid that are involved in numerous homeostatic biological functions and inflammation. They are generated by cyclooxygenase isozymes and 5-lipoxygenase, respectively, and their biosynthesis and actions are blocked by clinically relevant nonsteroidal anti-inflammatory drugs, the newer generation coxibs (selective inhibitors of cyclooxygenase-2), and leukotriene modifiers. The prime mode of prostaglandin and leukotriene action is through specific G protein-coupled receptors, many of which have been cloned recently, thus enabling specific receptor agonist and antagonist development. Important insights into the mechanisms of inflammatory responses, pain, and fever have been gleaned from our current understanding of eicosanoid biology. PMID- 11729304 TI - Lysophospholipids--receptor revelations. AB - Upon cell activation, membrane phospholipids are metabolized into potent lysophospholipid (LP) mediators, such as sphingosine 1-phosphate and lysophosphatidic acid. LPs fulfill signaling roles in organisms as diverse as yeast and humans. The recent discovery of G protein-coupled receptors for LPs in higher eukaryotes, and their involvement in regulating diverse processes such as angiogenesis, cardiac development, neuronal survival, and immunity, has stimulated growing interest in these lipid mediators. LP receptor biology has generated insights into fundamental cellular mechanisms and may provide therapeutic targets for drug development. PMID- 11729305 TI - Hedgehog signaling: a tale of two lipids. AB - Hedgehog proteins constitute one of the major classes of intercellular signals that control inductive interactions during animal development. These proteins undergo unusual lipid modifications and signal through an unconventional transmembrane protein receptor that is characterized by a sequence motif implicated in sterol sensing. Recent studies suggest that the lipid adducts regulate the range and potency of the signals, whereas the sterol-sensing domain is essential for receptor activity. PMID- 11729306 TI - Location, location, location: membrane targeting directed by PX domains. AB - Phosphoinositide (PI)-binding domains play critical roles in the intracellular localization of a variety of cell-signaling proteins. The 120-amino acid Phox homology (PX) domain targets proteins to organelle membranes through interactions between two conserved basic motifs within the PX domain and specific PIs. The combination of protein-lipid and protein-protein interactions ensures the proper localization and regulation of PX domain-containing proteins. Upon proper localization, PX domain-containing proteins can then bind to additional proteins and execute their functions in a diverse set of biological pathways, including intracellular protein transport, cell growth and survival, cytoskeletal organization, and neutrophil defense. PMID- 11729307 TI - Cloned cattle can be healthy and normal. PMID- 11729308 TI - Endangered right whales on the southeastern Bering Sea shelf. PMID- 11729309 TI - Delineation of mRNA export pathways by the use of cell-permeable peptides. AB - The transport of messenger RNAs (mRNAs) from the nucleus to the cytoplasm involves adapter proteins that bind the mRNA as well as receptor proteins that interact with the nuclear pore complex. We demonstrate the utility of cell permeable peptides designed to interfere with interactions between potential adapter and receptor proteins to define the pathways accessed by particular mRNAs. We show that HuR, a protein implicated in the stabilization of short-lived mRNAs containing AU-rich elements (AREs), serves as an adapter for c-fos mRNA export through two pathways. One involves the HuR shuttling domain, HNS, which exhibits a heat shock-sensitive interaction with transportin 2 (Trn2); the other involves two protein ligands of HuR-pp32 and APRIL-which contain leucine-rich nuclear export signals (NES) recognized by the export receptor CRM1. Heterokaryon and in situ hybridization experiments reveal that the peptides selectively block the nucleocytoplasmic shuttling of their respective adapter proteins without perturbing the overall cellular distribution of polyadenylated mRNAs. PMID- 11729310 TI - Photoinduced conversion of silver nanospheres to nanoprisms. AB - A photoinduced method for converting large quantities of silver nanospheres into triangular nanoprisms is reported. The photo-process has been characterized by time-dependent ultraviolet-visible spectroscopy and transmission electron microscopy, allowing for the observation of several key intermediates in and characteristics of the conversion process. This light-driven process results in a colloid with distinctive optical properties that directly relate to the nanoprism shape of the particles. Theoretical calculations coupled with experimental observations allow for the assignment of the nanoprism plasmon bands and for the first identification of two distinct quadrupole plasmon resonances for a nanoparticle. Unlike the spherical particles they are derived from that Rayleigh light-scatter in the blue, these nanoprisms exhibit scattering in the red, which could be useful in developing multicolor diagnostic labels on the basis not only of nanoparticle composition and size but also of shape. PMID- 11729311 TI - Total synthesis of ciguatoxin CTX3C. AB - More than 20,000 people suffer annually from ciguatera seafood poisoning in subtropical and tropical regions. The extremely low content of the causative neurotoxins, designated as ciguatoxins, in fish has hampered the isolation, detailed biological studies, and preparation of anti-ciguatoxin antibodies for detecting these toxins. The large (3 nanometers long) and complicated molecular structure of ciguatoxins has impeded chemists from completing their total synthesis. Our highly convergent strategic approach featuring the chemoselective ring-closing metathesis reaction as a key tactic has enabled the total synthesis of ciguatoxin CTX3C, which will provide a practical supply for further studies. PMID- 11729312 TI - Engineering crystal symmetry and polar order in molecular host frameworks. AB - A crystal design strategy is described that produces a series of solid-state molecular host frameworks with prescribed lattice metrics and polar crystallographic symmetries. This represents a significant advance in crystal engineering, which is typically limited to manipulation of only gross structural features. The host frameworks, constructed by connecting flexible hydrogen-bonded sheets with banana-shaped pillars, sustain one-dimensional channels that are occupied by guest molecules during crystallization. The polar host frameworks enforce the alignment of these guests into polar arrays, with properly chosen guests affording inclusion compounds that exhibit second harmonic generation because of this alignment. This protocol exemplifies a principal goal of modern organic solid-state chemistry: the precise control of crystal symmetry and structure for the attainment of a specific bulk property. PMID- 11729313 TI - Seismic detection of rigid zones at the top of the core. AB - Data from earthquakes in the Tonga-Fiji region recorded at a seismic array in northern Australia show evidence for rigid zones at the top of the outer core. The ScP waveforms can be modeled by thin (0.12 to 0.18 kilometer) zones of molten iron mixed with solid material with a small, but positive, S-wave velocity (0.6 to 0.8 kilometer per second) that enables the propagation of S-waves in the outermost core. The zones may be topographic highs of the core-mantle boundary filled by light core sediments and might be important for variation of Earth's nutation and for convection of the outer core. PMID- 11729314 TI - Detection of molecular hydrogen in the atmosphere of Mars. AB - Four hydrogen (H2) lines have been detected in a spectrum of Mars observed with the Far Ultraviolet Spectroscopic Explorer. Three of those lines are excited by the solar Lyman beta photons. The line intensities correspond to a column H2 abundance of 1.17 (+/-0.13) x 10(13) per square centimeter above 140 kilometers on Mars. A photochemical model for the upper atmosphere that simulates the observed H2 abundance results in an H2 mixing ratio of 15 +/- 5 parts per million in the lower atmosphere. The H2 and HD mixing ratios agree with photochemical fractionation of D (deuterium) between H2O and H2. Analysis of D fractionation among a few reservoirs of ice, water vapor, and molecular hydrogen on Mars implies that a global ocean more than 30 meters deep was lost since the end of hydrodynamic escape. Only 4% of the initially accreted water remained on the planet at the end of hydrodynamic escape, and initially Mars could have had even more water (as a proportion of mass) than Earth. PMID- 11729315 TI - Mediterranean Sea surface radiocarbon reservoir age changes since the last glacial maximum. AB - Sea surface reservoir ages must be known to establish a common chronological framework for marine, continental, and cryospheric paleoproxies, and are crucial for understanding ocean-continent climatic relationships and the paleoventilation of the ocean. Radiocarbon dates of planktonic foraminifera and tephra contemporaneously deposited over Mediterranean marine and terrestrial regions reveal that the reservoir ages were similar to the modern one (approximately 400 years) during most of the past 18,000 carbon-14 years. However, reservoir ages increased by a factor of 2 at the beginning of the last deglaciation. This is attributed to changes of the North Atlantic thermohaline circulation during the massive ice discharge event Heinrich 1. PMID- 11729316 TI - Effects of marine reserves on adjacent fisheries. AB - Marine reserves have been widely promoted as conservation and fishery management tools. There are robust demonstrations of conservation benefits, but fishery benefits remain controversial. We show that marine reserves in Florida (United States) and St. Lucia have enhanced adjacent fisheries. Within 5 years of creation, a network of five small reserves in St. Lucia increased adjacent catches of artisanal fishers by between 46 and 90%, depending on the type of gear the fishers used. In Florida, reserve zones in the Merritt Island National Wildlife Refuge have supplied increasing numbers of world record-sized fish to adjacent recreational fisheries since the 1970s. Our study confirms theoretical predictions that marine reserves can play a key role in supporting fisheries. PMID- 11729317 TI - Ecological meltdown in predator-free forest fragments. AB - The manner in which terrestrial ecosystems are regulated is controversial. The "top-down" school holds that predators limit herbivores and thereby prevent them from overexploiting vegetation. "Bottom-up" proponents stress the role of plant chemical defenses in limiting plant depredation by herbivores. A set of predator free islands created by a hydroelectric impoundment in Venezuela allows a test of these competing world views. Limited area restricts the fauna of small (0.25 to 0.9 hectare) islands to predators of invertebrates (birds, lizards, anurans, and spiders), seed predators (rodents), and herbivores (howler monkeys, iguanas, and leaf-cutter ants). Predators of vertebrates are absent, and densities of rodents, howler monkeys, iguanas, and leaf-cutter ants are 10 to 100 times greater than on the nearby mainland, suggesting that predators normally limit their populations. The densities of seedlings and saplings of canopy trees are severely reduced on herbivore-affected islands, providing evidence of a trophic cascade unleashed in the absence of top-down regulation. PMID- 11729318 TI - Matching spiracle opening to metabolic need during flight in Drosophila. AB - The respiratory exchange system of insects must maximize the flux of respiratory gases through the spiracles of the tracheal system while minimizing water loss. This trade-off between gas exchange and water loss becomes crucial when locomotor activity is increased during flight and metabolic needs are greatest. Insects that keep their spiracles mostly closed during flight reduce water loss but limit the flux of oxygen and carbon dioxide into and out of the tracheal system and thus attenuate locomotor performance. Insects that keep their spiracles completely open allow maximum gas exchange but face desiccation stress more quickly. Experiments in which water vapor was used as a tracer gas to track changes in the conductance of the respiratory system indicated that flying fruit flies minimize potential water loss by matching the area of the open spiracles to their gas exchange required for metabolic needs. This behavior maintained approximately constant pressure for carbon dioxide (1.35 kilopascals) and oxygen (19.9 kilopascals) within the tracheal system while reducing respirometric water loss by up to 23% compared with a strategy in which the spiracles are held wide open during flight. The adaptive spiracle-closing behavior in fruit flies has general implications for the ecology of flying insects because it shows how these animals may cope with environmental challenges during high locomotor performance. PMID- 11729319 TI - Real-time single-molecule imaging of the infection pathway of an adeno-associated virus. AB - We describe a method, based on single-molecule imaging, that allows the real-time visualization of the infection pathway of single viruses in living cells, each labeled with only one fluorescent dye molecule. The tracking of single viruses removes ensemble averaging. Diffusion trajectories with high spatial and time resolution show various modes of motion of adeno-associated viruses (AAV) during their infection pathway into living HeLa cells: (i) consecutive virus touching at the cell surface and fast endocytosis; (ii) free and anomalous diffusion of the endosome and the virus in the cytoplasm and the nucleus; and (iii) directed motion by motor proteins in the cytoplasm and in nuclear tubular structures. The real-time visualization of the infection pathway of single AAVs shows a much faster infection than was generally observed so far. PMID- 11729320 TI - Physiological migration of hematopoietic stem and progenitor cells. AB - Hematopoietic stem cells (HSCs) reside predominantly in bone marrow, but low numbers of HSCs are also found in peripheral blood. We examined the fate of blood borne HSCs using genetically marked parabiotic mice, which are surgically conjoined and share a common circulation. Parabionts rapidly established stable, functional cross engraftment of partner-derived HSCs and maintained partner derived hematopoiesis after surgical separation. Determination of the residence time of injected blood-borne progenitor cells suggests that circulating HSCs/progenitors are cleared quickly from the blood. These data demonstrate that HSCs rapidly and constitutively migrate through the blood and play a physiological role in, at least, the functional reengraftment of unconditioned bone marrow. PMID- 11729321 TI - T cell responses modulated through interaction between CD8alphaalpha and the nonclassical MHC class I molecule, TL. AB - The thymus leukemia antigen (TL) is a nonclassical class I molecule, expressed abundantly on intestinal epithelial cells. We show that, in contrast to other major histocompatibility complex (MHC) class I molecules that bind CD8alphabeta, TL preferentially binds the homotypic form of CD8alpha (CD8alphaalpha). Thus, TL tetramers react specifically to CD8alphaalpha-expressing cells, including most intestinal intraepithelial lymphocytes. Compared with CD8alphabeta, which recognizes the same MHC as the T cell receptor (TCR) and thus acts as a TCR coreceptor, high-affinity binding of CD8alphaalpha to TL modifies responses mediated by TCR recognition of antigen presented by distinct MHC molecules. These findings define a novel mechanism of lymphocyte regulation through CD8alphaalpha and MHC class I. PMID- 11729322 TI - RGS-PX1, a GAP for GalphaS and sorting nexin in vesicular trafficking. AB - Heterotrimeric GTP-binding proteins (G proteins) control cellular functions by transducing signals from the outside to the inside of cells. Regulator of G protein signaling (RGS) proteins are key modulators of the amplitude and duration of G protein-mediated signaling through their ability to serve as guanosine triphosphatase-activating proteins (GAPs). We have identified RGS-PX1, a Galpha(s)-specific GAP. The RGS domain of RGS-PX1 specifically interacted with Galpha(s), accelerated its GTP hydrolysis, and attenuated Galpha(s)-mediated signaling. RGS-PX1 also contains a Phox (PX) domain that resembles those in sorting nexin (SNX) proteins. Expression of RGS-PX1 delayed lysosomal degradation of the EGF receptor. Because of its bifunctional role as both a GAP and a SNX, RGS-PX1 may link heterotrimeric G protein signaling and vesicular trafficking. PMID- 11729323 TI - Phosphatidic acid-mediated mitogenic activation of mTOR signaling. AB - The mammalian target of rapamycin (mTOR) governs cell growth and proliferation by mediating the mitogen- and nutrient-dependent signal transduction that regulates messenger RNA translation. We identified phosphatidic acid (PA) as a critical component of mTOR signaling. In our study, mitogenic stimulation of mammalian cells led to a phospholipase D-dependent accumulation of cellular PA, which was required for activation of mTOR downstream effectors. PA directly interacted with the domain in mTOR that is targeted by rapamycin, and this interaction was positively correlated with mTOR's ability to activate downstream effectors. The involvement of PA in mTOR signaling reveals an important function of this lipid in signal transduction and protein synthesis, as well as a direct link between mTOR and mitogens. Furthermore, these studies suggest a potential mechanism for the in vivo actions of the immunosuppressant rapamycin. PMID- 11729324 TI - Regulation of cell survival by secreted proneurotrophins. AB - Neurotrophins are growth factors that promote cell survival, differentiation, and cell death. They are synthesized as proforms that can be cleaved intracellularly to release mature, secreted ligands. Although proneurotrophins have been considered inactive precursors, we show here that the proforms of nerve growth factor (NGF) and the proforms of brain derived neurotrophic factor (BDNF) are secreted and cleaved extracellularly by the serine protease plasmin and by selective matrix metalloproteinases (MMPs). ProNGF is a high-affinity ligand for p75(NTR) with high affinity and induced p75NTR-dependent apoptosis in cultured neurons with minimal activation of TrkA-mediated differentiation or survival. The biological action of neurotrophins is thus regulated by proteolytic cleavage, with proforms preferentially activating p75NTR to mediate apoptosis and mature forms activating Trk receptors to promote survival. PMID- 11729325 TI - Lobster sniffing: antennule design and hydrodynamic filtering of information in an odor plume. AB - The first step in processing olfactory information, before neural filtering, is the physical capture of odor molecules from the surrounding fluid. Many animals capture odors from turbulent water currents or wind using antennae that bear chemosensory hairs. We used planar laser-induced fluorescence to reveal how lobster olfactory antennules hydrodynamically alter the spatiotemporal patterns of concentration in turbulent odor plumes. As antennules flick, water penetrates their chemosensory hair array during the fast downstroke, carrying fine-scale patterns of concentration into the receptor area. This spatial pattern, blurred by flow along the antennule during the downstroke, is retained during the slower return stroke and is not shed until the next flick. PMID- 11729326 TI - Modulation of presbycusis: current status and future directions. AB - Literature and ideas are reviewed concerning the modulation of presbycusis - the influence of variables that can alter the severity and/or time course of presbycusis or counteract its negative aspects. Eleven topics are identified: variables related to biological aging; genetics; noise-induced hearing loss; moderately augmented acoustic environment; neural plasticity and the central auditory system; neural plasticity and hearing aids; socioeconomic and cultural barriers to hearing aid use; lifestyle (diet, exercise, etc.); medical variables; pharmaceutical interventions for presbycusis, and cognitive variables. It is concluded that the field of otogerontology will best be served by a comprehensive, integrative interaction among basic researchers and clinical scientists who will continue to learn how the auditory problems associated with presbycusis can be intentionally modulated in beneficial ways. PMID- 11729327 TI - A novel microperfusion system for the long-term local supply of drugs to the inner ear: implantation and function in the rat model. AB - Local therapy is practiced for middle and inner ear diseases but is usually restricted to cases of ear drum perforation or repeated invasive intratympanic drug application. Perfusion of drugs on the round window or through the scalae of animals using a pump system suggests that the chronic local drug treatment might also be feasible in humans. However, drug delivery systems that are currently on the market involve repeated reimplantation if they are to be used for long-term drug supply. A bone-anchored, totally implantable micro-drug delivery system (MDS) for patient-controlled drug supply has been developed [Lehner et al., 1997]. In this study, we show the first successful long-term in vivo test of the MDS micro-pump in rats. The process of implantation and first functional tests will be described. The biomaterial used to manufacture the delivery system did not cause any inflammation reaction in any of the 9 animals successfully implanted. After activation of the micro-pump, the drug reservoir and port was found to be fluid-tight. Bolus applications of tetrodotoxin (TTX) to the round window induced a transient decrease of evoked brainstem responses. In 2 animals which carried the MDS for more than 8 months the proper functioning of the pumping device was examined in a 2-3 week interval over a 3 month period. The MDS can be autoclaved even after long-term implantation and can then be reused for subsequent implantations. Designed for life-long implantation in humans, the demonstration of an effective long-term drug supply to the inner ear using the MDS provides an encouraging first step towards future long-term drug treatment of the inner ear in humans. PMID- 11729328 TI - HSV-1 not only in human vestibular ganglia but also in the vestibular labyrinth. AB - Reactivation of herpes simplex virus type 1 (HSV-1) in the vestibular ganglion (VG) is the suspected cause of vestibular neuritis (VN). Recent studies reported the presence of HSV-1 DNA not only in human VGs but also in vestibular nuclei, a finding that indicates the possibility of viral migration to the human vestibular labyrinth. Distribution of HSV-1 DNA was determined in geniculate ganglia, VGs, semicircular canals, and macula organs of 21 randomly obtained human temporal bones by nested PCR. Viral DNA was detected in 48% of the labyrinths, 62% of the VGs, and 57% of the geniculate ganglia. The potential significance of this finding is twofold: (1) Inflammation in VN could also involve the labyrinth and thereby cause acute unilateral vestibular deafferentation. (2) As benign paroxysmal positional vertigo often occurs in patients who have had VN, it could also be a sequel of viral labyrinthitis. PMID- 11729329 TI - Latency of auditory evoked field deflection N100m ruled by pitch or spectrum? AB - The auditory evoked field (AEF) in response to pure tones of 250 and 1000 Hz and a complex tone with a periodicity of 4 ms (composed of the frequencies 1000, 1250, 1500, 1750, and 2000 Hz), corresponding to a pitch of 250 Hz, was recorded with a 37-channel neuromagnetometer system. The intensity was 60 dB sensation level (SL). Two different stimulus durations were examined in 12 subjects: 500 ms (long tones) and 100 ms (short tones). The stimulus onset asynchrony (SOA) was uniformly distributed between 3 and 4 s for the long tones and between 0.8 and 1.2 s for the short tones. Each subject was investigated four times, to assess the intraindividual variability. The mean latency of the AEF deflection N100m turned out to be similar for the long and the short tones: about 98 and 87 ms for the pure tones of 250 Hz and 1000 Hz, respectively, and 95 ms for the complex tone with a pitch of 250 Hz. However, a great interindividual variability was observed, exhibiting no consistent relationship between the N100m latencies for the three different tones, except that the response to the pure tone of 1000 Hz generally occurred earlier. In conclusion, this study does not support the proposal that the N100m latency represents a code for pitch, although a low pitch appears to be a factor favoring a longer N100m latency. PMID- 11729330 TI - The low and high frequency auditory steady state responses mature at different rates. AB - The age-related changes in the fast rate (70-110 Hz) auditory steady state response elicited by multiple-frequency tones (MSSR) that were amplitude modulated (AM) are reported here. The MSSR was recorded in a sample of 64 well babies distributed into three age groups: 0-29 days (n = 25); 1-6 months (n = 26); 7-12 months (n = 13). Four simultaneously presented AM tones (0.5, 1, 2 and 4 kHz) were delivered monaurally through TDH 49 earphones, at different intensities (between 90 and 30 dB SPL). Clear developmental changes were found between birth and 12 months of age in response threshold, amplitude and detectability. Statistical analysis revealed that these changes occurred at different rates for low- and high- frequency responses. Nonetheless adult-like hearing thresholds were estimated reasonably well in most newborns and well babies, for all frequencies tested. Therefore the MSSR technique could be useful for objective frequency-specific audiometry beginning at birth. PMID- 11729331 TI - Language acquisition in young German-speaking children with cochlear implants: individual differences and implications for conceptions of a 'sensitive phase'. AB - The acquisition of language was studied longitudinally in a sample of 22 German speaking children with cochlear implants (mean implantation age 29 months) and a control group of 22 normally hearing children. Spontaneous speech samples were collected over 27-36 months, starting at the the one-word stage. Results indicate that grammatical progress as measured by mean length of utterance was slower for cochlear-implanted children. However, there were substantial individual differences in the cochlear-implanted group. While 10 cochlear-implanted children progressed at pace with normally hearing children, 12 cochlear-implanted children remained well behind. Cochlear-implanted children who showed fast progress at an early stage continued to make fast progress as time went on, and those who showed slow progress early on continued to progress slowly. Pre-operative hearing was a better predictor of subsequent linguistic growth than age at implantation. Increases in vocabulary were associated with grammatical progress in both groups. PMID- 11729332 TI - Efficacy of every-other-day administration of conjugated equine estrogen and medroxyprogesterone acetate on gonadotropin-releasing hormone agonists treatment in women with endometriosis. AB - We performed a randomized controlled study to determine the efficacy of add backed therapy by every-other-day administration of 0.625 mg conjugated equine estrogen (CEE) and 2.5 mg medroxyprogesterone acetate (MPA) on GnRH agonists (GnRH-a) treatment in Japanese women with symptomatic endometriosis. At the end of treatment, serum estrone and estradiol levels in the add-back group (n = 11) were significantly higher than those in the control group (n = 10). The assessment of Beecham classification by bimanual examination, serum CA-125 levels, and the frequency of genital bleeding revealed no significant differences between the two groups. The add-back group showed reduced Kupperman indices relative to those of the control group, and could prevent the loss of bone density. These findings led to a conclusion that GnRH-a therapy added back by every-other-day administration of 0.625 mg CEE and 2.5 mg MPA was a safe and effective treatment for Japanese women with endometriosis. PMID- 11729333 TI - Clinical features affecting the results of estrogen replacement therapy on bone density in Japanese postmenopausal women. AB - Although estrogen replacement therapy (ERT) has been established as an effective treatment for postmenopausal bone loss, the clinical features which predict the effects of ERT have not been well investigated in Japanese postmenopausal women. We analyzed the role of physical factors influencing the effect of ERT on vertebral bone mineral density (BMD) in 94 Japanese postmenopausal women treated for 2 years or longer. The increase in BMD with ERT is 17.6 +/- 27.6 mg/cm(2)/year (mean +/- SD) during the first 2 years. Rates of BMD change were negatively correlated with the estimated initial BMD, and positively correlated with age and years since menopause, while no correlation was noted with the body mass index by a simple correlation analysis. The relationships between BMD change and estimated initial BMD or age also held in a multiple regression analysis. The estimated initial BMD and age together accounted for 34.4% of the BMD change during ERT. Furthermore, there were very few (2.4%) nonresponders with a negative linear regression slope of BMD in the osteoporosis and osteopenia group, although 32.7% of the normal initial BMD group were nonresponders. These results suggest that the initial BMD and age are potent predictive factors of the ERT effect on BMD change in Japanese postmenopausal women. PMID- 11729334 TI - Risk and prediction of preterm delivery in pregnancies complicated by antepartum hemorrhage of unknown origin before 34 weeks. AB - OBJECTIVE: To assess the timing and risk factors of preterm delivery in pregnancies complicated by antepartum hemorrhage of unknown origin (APHUO) before 34 weeks of gestation, and to formulate a clinical management protocol. METHODS: A retrospective study involving singleton pregnancies with APHUO occurring before the 34th week of gestation was performed over a 4-year period at a university teaching hospital. RESULTS: Seventy-five cases were identified. Fifty-nine (78.7%) did not have any associated uterine contractions, 74.7 and 90.7% of the cases were classified as having mild bleeding according to history and physical examination, respectively. The number of cases delivering before 37 weeks of gestation and before 34 weeks were 28 (37.3%) and 22 (29.3%), respectively. Eighteen cases (24%) delivered within the first week of APHUO, and 4 (5.3%) within the first day. The number of cases delivering within the first week was significantly higher when there were uterine contractions (62.5 vs. 13.6%, p < 0.001). The severity of bleeding was not shown to be related to the time of delivery. CONCLUSION: APHUO that occurs before 34 weeks is associated with a high risk of preterm delivery before 34 weeks (29.3%). The chance of delivery within the first week is 62.5% when there are coexisting uterine contractions. Even if contractions are absent, the risk is still high (13.6%) and persists beyond the first week. We suggest that this group of patients should be managed with a course of steroid injection and hospitalization for 4 days. PMID- 11729335 TI - Pregnancy-associated changes in peripheral blood lymphocyte subpopulations in normal Kuwaiti women. AB - It has recently been reported that healthy pregnancy is associated with systemic immunosuppression. The aim of this study was to evaluate the numbers and distribution of lymphocyte subpopulations in normal, healthy pregnant Kuwaiti women. Thirty-four healthy normotensive women in the 3rd trimester of pregnancy were studied using flow cytometry to define lymphocyte subpopulations and were compared with 16 non-pregnant women. A decrease in the absolute numbers of lymphocytes was observed affecting T cells (CD3+, CD4+, CD8+), B cells (CD19+), and natural killer cells (CD16+/CD56+). When analyzed as a percentage of the total lymphocyte population, there was a significant decrease in B cells and an increase in CD4+ T cells. The T cell population revealed increased expression of CD25 on CD4+ and CD8+ cells, of HLA-DR on CD8+ cells, and of CD54 on CD4+ T cells. The reduced number of lymphocytes suggests that Kuwaiti females may be immunosuppressed in the 3rd trimester of pregnancy. The presence of activated CD4+ T cells could indicate the expression of a regulatory suppressor T cell population, as Treg cells are CD4+CD25+, and suppressor T cells are thought to be CD8+. Future work is required to explore the significance of these T cell populations in pregnancy. PMID- 11729336 TI - Circulating divalent cations in asymptomatic ovarian hyperstimulation and in vitro fertilization patients. AB - It is known that ovarian hyperstimulation and in vitro fertilization are accompanied by a steady increase in circulating estrogen and progesterone far beyond what is normal for young women. We have recently demonstrated that the biologically active fractions of calcium and magnesium in blood are altered depending on when in the menstrual phase a blood sample is drawn in normal cycling women. The serum ionized Ca/Mg ratio is also altered in accordance with the menstrual cycles. This suggests that the sex steroid hormones may modulate serum levels of ionized Mg and the ionized Ca/Mg ratio. We therefore studied the relationships between sex steroid hormones and the concentrations of ionized magnesium and calcium in the blood of hyperstimulated patients. We were able to demonstrate that with each increment in estrogen, a decrease in ionized Mg occurred, and as the progesterone rose in the blood, the ionized Ca/Mg ratio increased. Our results support the idea that serum estrogen and progesterone levels in women modulate the blood levels of circulating ionized Mg and the serum ionized Ca/Mg ratio. PMID- 11729337 TI - Serum leptin and somatotropin components correlate with neonatal birth weight. AB - OBJECTIVE: To determine whether cord sera leptin and components of the somatotropin axis - growth hormone (GH), total (t) and free (f) insulin-like growth factor (IGF), IGF-binding protein-3 (IGFBP-3), and insulin - correlate with birth weight. DESIGN: Cross-sectional study of 22 newborns, 12 with normal birth weight (NBW) and 10 with low birth weight (LBW), in a population of healthy mothers with an apparent normal pregnancy. METHODS: Paired mother-neonate blood samples were obtained at vaginal delivery in order to measure leptin and the somatotropin axis components. RESULTS: In all cases maternal blood concentrations of leptin, t and fIGF-I, its carrier protein IGFBP-3, and insulin were higher than in the cord sera of the newborns, regardless of their birth weight. On the contrary, maternal GH levels were lower than in their neonates. LBW neonates had decreased levels of leptin, tIGF-I, and IGFBP-3 as compared with those levels in NBW offspring; however, GH concentrations were higher in LBW neonates. Birth weight showed a significant correlation with cord sera leptin, tIGF-I, IGFBP-3, and GH; nevertheless birth weight was neither interrelated with fIGF-I nor with insulin levels. CONCLUSION: These data demonstrate that birth weight is significantly correlated with both leptin and some components of the somatotropin axis; on the other hand, no correlation was observed between leptin concentrations and each one of the components of the somatotropin axis. It is suggested that fetal leptin and the somatotropin axis cooperate in intrauterine growth and birth weight. PMID- 11729338 TI - Prevalence of human papillomavirus in elderly women with cervical cancer. AB - To investigate the relation between the prevalence of human papillomavirus (HPV) and age in cervical cancer patients, material from 93 patients with Ia-IIb cervical carcinoma was analyzed for the presence of HPV by both type-specific and general primer polymerase chain reaction. Patients were divided into 2 groups: 64 years or younger, and 65 years and older. There was no statistically significant difference in either the prevalence of HPV DNA or distribution of genotypes amongst the 2 groups. Therefore, HPV detection can be equally well used in the management and follow-up of elderly cervical cancer patients. PMID- 11729339 TI - Effect of the supine position on uterine and umbilical blood flow during the third trimester of uncomplicated pregnancies in multiparous patients. AB - We established the effects of the supine position on umbilical blood flow when measured during the third trimester in 30 multiparous, normotensive patients. Blood flow in the umbilical and uterine arteries and blood pressure in the brachial and popliteal arteries were blindly taken by two different observers: first in the lateral and 5 min later in the supine position. There was a significant difference in mean blood pressure between the two postures. However, there was no statistical difference in the pulse pressure or in systolic/diastolic ratio in the umbilical and uterine arteries between the two positions. Postural changes in normotensive multiparous patients do not affect uterine and umbilical blood flow during the third trimester of uncomplicated pregnancies. PMID- 11729340 TI - Localization of hyaluronan with a hyaluronan-specific hyaluronic acid binding protein in the placenta in pre-eclampsia. AB - Hyaluronan (HA), a high molecular weight polysaccharide, is a major component of connective tissue and is thus present in the extracellular matrix of most tissues. Increased serum concentrations have been reported in association with pre-eclampsia and liver malfunction, amongst other disorders. We have performed histochemical investigations with a HA-specific hyaluronic acid binding protein in placentas from uncomplicated pregnancies and from patients with pre-eclampsia. Staining for HA was found in the stroma and blood vessel walls of stem villi in all the placentas investigated. The syncytiotrophoblast and cytotrophoblast cells usually remained unstained. In addition, reactivity for HA was found within and on the surface of intervillous and perivillous fibrinoid deposits. Since fibrinoid deposits are increased in pre-eclampsia, our findings suggest that the increased HA serum concentrations in cases of pre-eclampsia could result from the stroma of the infarcted villi and from the fibrinoid deposits. HA may reach the maternal blood through fibrinoid gaps. PMID- 11729341 TI - Relation between plasma endothelin 1 levels and T helper 1: T helper 2 cell immunity in women with preeclampsia. AB - The aim of this study was to investigate the relationship between plasma endothelin 1 (ET-1) levels and T helper (Th)-1:Th2 cell immunity in women with preeclampsia. The percentage of Th1 and Th2 cells and the Th1:Th2 cell ratios in peripheral blood from 11 normal pregnant women and 11 patients with preeclampsia at 29-34 weeks of gestation were calculated using flow cytometry. The plasma ET-1 level was also determined using a modified radioimmunoassay. The plasma ET-1 concentrations and the Th1:Th2 cell ratios in normal pregnancies were significantly lower than those in patients with preeclampsia. Negative correlations were found between plasma ET-1 levels and Th2 cells in both the preeclamptic pregnancy groups and in the normal pregnant women. Our results indicate that elevated ET-1 levels are associated with a Th1:Th2 imbalance in preeclampsia. PMID- 11729342 TI - Hormone-related factors associated with hormone receptor levels in breast cancer. AB - Estrogen receptor (ER) and progesterone receptor (PgR) levels were determined in patients with a ductal infiltrating breast carcinoma. In premenopausal patients, the distribution of the PgR was higher than that of the ER. With increasing age of the patients, the concentration of the ER increased, whereas the PgR values showed an inverse relationship. According to the menstrual status, ER and PgR values were found to be higher in postmenopausal patients than in premenopausal ones. In relation to body fat distribution, in premenopausal patients the ER or PgR values were not found to be statistically significantly different (data no shown). In postmenopausal patients, the concentration of ER was diminished by 20% in patients with obesity of the upper body segment and by 40% in patients with obesity of the lower body segment as compared with the ER values of nonobese patients. While the PgR values diminished by 20% in patients with obesity of the lower body segment, in relation to PgR values of patients with obesity of the upper body segment, we found no statistically significant differences when compared with the PgR values of nonobese patients. With regard to the body mass index, this has remained constant in both types of body fat distribution. When body mass index values were compared with serum ER and PgR levels, no statistically significant differences were found. PMID- 11729343 TI - Parathion, a cholinesterase-inhibiting plaguicide induces changes in tertiary villi of placenta of women exposed: a scanning electron microscopy study. AB - The objective of this work was to describe the anatomy of placentas from women who were at risk of exposure to parathion during their pregnancy, when examined with the light and scanning electron microscopes. Twenty term placentas were analyzed; 10 from women living in an agricultural area, who were at risk of exposure to parathion during their pregnancy, and 10 from women living in an urban area, not expressly exposed to pesticides. Each sample was examined with both light and scanning electron microscopes. Cholinesterase activity was significantly reduced in blood from women of the exposed group. In some placentas of women exposed to parathion, recent microinfarctions, microcalcifications and increased deposition of fibrinoid material were seen, along with a larger proportion of atypical characteristics of villi, such as bullous and balloon-like formations with nonhomogeneous surface, and other areas devoid of microvilli. These observations suggest that in chronic exposure to pesticides, the rate of atypical characteristics of placental villi increases, which could be related to changes in the fetus biology. In this study, one newborn from the exposed group showed intrauterine growth retardation and another one, some signs of hypoxia. PMID- 11729344 TI - A perforated sigmoid diverticulum abscess difficult to differentiate from an ovarian tumor. AB - We report a case of a perforated sigmoid diverticulum abscess that was difficult to differentiate from an ovarian tumor. A 53-year-old woman was diagnosed with an ovarian tumor, but laparotomy revealed a perforated sigmoid diverticulum abscess and appendicitis, with normal ovaries. PMID- 11729345 TI - Blood flow after intraarterial infusion chemotherapy in a patient with advanced cervical cancer. AB - We presented a case of locally advanced cervical cancer treated by intraarterial infusion chemotherapy, and evaluated the blood flow of uterine arteries before and after chemotherapy by using a transvaginal ultrasonic color Doppler device. Pulsatility index (PI) of each uterine artery increased after first course of chemotherapy compared to that of before chemotherapy. But PI did not change after second course in spite of a significant reduction in tumor size. Blood flow change assessed by Doppler ultrasound may be a limited but useful parameter for the efficacy of neoadjuvant chemotherapy in patients with cervical cancer. PMID- 11729346 TI - The role of interleukin 10 in the regulation of allergic immune responses. AB - Several clinical studies and animal models have shown that Th2 lymphocytes play a key role in the pathophysiology of IgE-mediated allergic immune responses like allergic rhinitis and asthma or venom anaphylaxis. Classical specific immunotherapy (SIT) that has been proven to be clinically effective can serve as a role model for immunological changes that are associated with amelioration of allergic diseases. During SIT, the Th2-dominated immune response is modified towards a Th1 response leading to a decline in allergen-specific IgE and an increase in allergen-specific IgG production. Most importantly, however, production of the immunosuppressive/-regulatory cytokine interleukin 10 (IL-10) is also induced leading to T cell tolerance and prevention of tissue inflammation. In this article the role of IL-10-producing T cells in the regulation of allergic immune responses will be discussed. PMID- 11729347 TI - How to deal with polarized Th2 cells: exploring the Achilles' heel. AB - The central effector cells in the pathogenesis of atopic allergic diseases are type 2 T helper (Th2) cells, which display an aberrant cytokine profile dominated by type 2 cytokines. Initial reports from mouse studies indicated that established and committed Th2 cells are stable and unsusceptible to modulation. However, there is a growing awareness that in humans, established effector Th2 cells are more flexible and can be reverted to predominant Th1 phenotypes. In fact, the Th1-driving cytokine interleukin (IL)-12 is the crucial factor in this respect. IL-12 is mainly produced by dendritic cells (DC), which can be primed for high or low IL-12 production, depending on inflammatory and/or microbial signals they encounter during their residence in the peripheral tissues. Accordingly, both the regulation of and the priming for IL-12 production in DC form ideal targets for therapeutic intervention. The development of new therapies for atopic allergy now focuses on local IL-12-promoting substances to target both the development of new Th2 cells and the persistent population of established allergen-specific Th2 cells. PMID- 11729348 TI - IgE and IgG binding epitopes on alpha-lactalbumin and beta-lactoglobulin in cow's milk allergy. AB - BACKGROUND: Cow's milk is one of the most common causes of food allergy in the first years of life. We recently defined IgE and IgG binding epitopes for alpha(s1)-casein, a major cow's milk allergen, and found an association between recognition of certain epitopes and clinical symptoms of cow's milk allergy (CMA). Since alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) are suspected to be significant allergens in cow's milk, we sought to determine the structure of sequential epitopes recognized by IgE antibodies to these proteins. We further sought to assess the pattern of epitope recognition in association with the clinical outcome of CMA. METHODS: According to the known amino acid sequence of ALA and BLG, 57 and 77 overlapping decapeptides (offset by two amino acids), respectively, were synthesized on a cellulose derivatized membrane. Sera from 11 patients 4-18 years of age with persistent CMA (IgE to cow's milk >100 kU(A)/l) were used to identify IgE binding epitopes. In addition, 8 patients < 3 years of age and likely to outgrow their milk allergy (IgE to cow's milk < 30 kU(A)/l) were used to investigate the differences in epitope recognition between patients with 'persistent' and those with 'transient' CMA. Seven patients 4-18 years of age were used for assessing the IgG binding regions. RESULTS: In patients with persistent allergy, four IgE binding and three IgG binding regions were identified on ALA, and seven IgE and six IgG binding epitopes were detected on BLG. The younger patients that are likely to outgrow their allergy recognized only three of these IgE binding epitopes on BLG and none on ALA. CONCLUSIONS: The presence of IgE antibodies to multiple linear allergenic epitopes may be a marker of persistent CMA. The usefulness of IgE binding to distinct epitopes on whey proteins in defining the patients that would have a lifelong CMA needs to be investigated in further studies. PMID- 11729349 TI - Eotaxin-1-deficient mice develop airway eosinophilia and airway hyperresponsiveness. AB - BACKGROUND: The accumulation of eosinophils in the lung is a hallmark of asthma. In addition to cytokines such as IL-5 which are essential, chemokines have been implicated in the recruitment of eosinophils to the airway. In particular, eotaxin has been shown to be a selective and potent eosinophil chemoattractant, important in the pathogenesis of allergic disease. The goal of the present study was to define the role of eotaxin-1 in the development of allergen-induced eosinophilic airway inflammation and airway hyperresponsiveness (AHR) to inhaled methacholine (MCh). METHODS: Eotaxin-1-deficient mice were sensitized and exposed to a single challenge with allergen. Airway function and airway and tissue as well as peripheral blood and bone marrow eosinophilia were examined 18 and 48 h after the last challenge. RESULTS: Following allergen sensitization and challenge, eotaxin-1-deficient mice developed levels of AHR to inhaled MCh at 18 and 48 h comparable to controls. Further, levels of bronchoalveolar lavage (BAL) and tissue eosinophilia at the same time points were comparable in the two strains of mice. Tissue eosinophilia, assessed by quantitating major basic protein staining cells, preceded BAL eosinophilia in a similar manner. Bone marrow and peripheral blood eosinophilia were unimpaired in deficient mice. CONCLUSION: The results demonstrate that the major eotaxin, eotaxin-1 is not essential for the development of airway eosinophilia or AHR, implying that other chemokines, alone or in combination, can overcome this deficiency. PMID- 11729350 TI - Prolonged antigen exposure ameliorates airway inflammation but not remodeling in a mouse model of bronchial asthma. AB - BACKGROUND: In naive rodents, repeated exposure to aerosolized antigen induces suppression of the Th2 response to the antigen. We hypothesized that more prolonged exposure of established asthma model to antigen aerosols may downregulate asthmatic phenotype. METHODS: After establishing an ovalbumin (OVA) induced asthma model, mice were further exposed to OVA (prolonged exposure group) or phosphate-buffered saline (positive controls) 3 days per week for 6 weeks. During week 7, the mice of both groups were finally challenged with OVA. RESULTS: Prolonged OVA exposure resulted in marked suppression of serum OVA-specific immunoglobulin E (IgE) antibody levels, eosinophilia of the airway, and airway hyperresponsiveness (AHR). However, airway remodeling characterized by goblet cell hyperplasia and airway fibrosis was observed to the same degree in both groups. These effects were accompanied by diminished production of Th2 cytokines such as interleukin-4 (IL-4), IL-5 and IL-13 in bronchoalveolar lavage fluid (BALF) and cultured supernatant of splenocytes. Furthermore, prolonged exposure markedly increased IL-12 levels in BALF. CONCLUSIONS: Prolonged antigen exposure has inhibitory effects on eosinophilic inflammation, AHR and IgE response to antigen, but not on airway remodeling, presumably via inhibition of Th2 cytokines and increased IL-12 production in the lungs. PMID- 11729351 TI - A Th1-inducing adjuvant, MPL, enhances antibody profiles in experimental animals suggesting it has the potential to improve the efficacy of allergy vaccines. AB - BACKGROUND: Monophosphoryl lipid A (MPL) is a detoxified derivative of the lipopolysaccharide (LPS) moiety of Salmonella minnesota R595, which has retained immunostimulatory activities. MPL has been administered to many subjects in clinical trials as an adjuvant component of infectious disease vaccines and is currently a component of a licensed cancer vaccine, Melacine (Corixa Inc., Schering Plough). MPL has, in particular, been shown to promote Th1-type antigen specific responses. L-tyrosine is a depot adjuvant which is fully metabolisable and has been successfully employed in allergy vaccines for a number of years. METHODS: Mice were immunised with MPL adjuvant in conjunction with separate preparations of either ovalbumin or glutaraldehyde-modified ragweed pollen extract both coprecipitated with L-tyrosine. The specific antibody isotypes IgG1, IgG2a, IgG2b and also IgE were measured. Rats received booster injections of keyhole limpet haemocyanin (KLH) in conjunction with MPL adjuvant following priming with KLH in alum alone. KLH-specific antibody responses were measured. RESULTS: It was shown that a combination of L-tyrosine and MPL were synergistic in enhancing murine antigen specific IgG antibody responses without enhancing antigen specific IgE responses. Furthermore, this adjuvant combination promoted strong IgG2 antigen specific responses indicative of a Th1 directed response. In KLH sensitised rats, treatment with MPL was shown to prevent a secondary IgE antibody response when injected with booster injections of antigen. CONCLUSIONS: Immunisation of mice with two different antigens adsorbed to L-tyrosine induced a Th1-skewed primary response when in conjunction with MPL adjuvant which also generally enhances a specific IgG response. Incorporation of MPL adjuvant in the immunisation of rats prevented a secondary specific IgE response. These results suggest that the employment of this new adjuvant in clinical allergy vaccination formulations may result in an improved efficacy which could be utilised in various ways to improve compliance. PMID- 11729352 TI - Roles of superoxide dismutase in rat mast cell granules. AB - BACKGROUND: It has been suggested that in the granules of rat mast cells there is some kind of superoxide dismutase (SOD), but details of this SOD in mast cells remain unclear. In the present study, we studied the mode of existence of SOD in mast cells and its releasing mechanism from the granules. In addition, we discussed the physiological role of SOD in allergic events. METHODS: Purified rat mast cells were disrupted with a sonic disrupter and granules (sample I) were separated from supernatant (sample II) by centrifugation. The granules were treated with 1 mM Ca(2+), and the supernatant (sample III) was separated from the pellet (sample IV). Sample III was applied to a heparin column and the eluate was used as sample V. SOD activity was measured in these samples. RESULTS: SOD existed in mast cell granules as a heparin-binding and an inactive form. However, when granules were released and exposed to high Ca(2+) concentration, SOD was discharged from heparin and shifted to the active form. The expression of macrophage inflammatory protein-1 alpha mRNA was enhanced when hydrogen peroxide (H(2)O(2)) or sample III with the xanthine-xanthine oxidase system were added to the culture media. CONCLUSIONS: These findings suggest that in stimulated rat mast cells, the released SOD may transform the generated superoxide anion into H(2)O(2), and the generated H(2)O(2) may enhance the expression of chemokine mRNA in the mast cells. PMID- 11729353 TI - Critical role for caspases 3 and 8 in neutrophil but not eosinophil apoptosis. AB - BACKGROUND: Apoptosis is a necessary process to control cell numbers in multicellular organisms. In many chronic inflammatory diseases, reduced cell death of different types of granulocytes is one important mechanism for cell accumulation. Here, we studied the role of caspases in neutrophil and eosinophil apoptosis in the presence or absence of granulocyte-macrophage stimulating factor and anti-CD95 monoclonal antibodies, respectively. METHODS: Granulocytes were isolated from human blood using standard protocols. Immunoblot and functional studies with cell-permeable specific peptide inhibitors were performed to analyze caspase involvement. Fas receptor and Fas ligand expression was analyzed by RT PCR, flow cytometry, and immunoblotting. Cell death was analyzed by ethidium bromide exclusion test. RESULTS: Caspases 3 and 8 are critically involved in the regulation of neutrophil apoptosis in vitro. In contrast, these two caspases did not appear to play a major role in the regulation of eosinophil apoptosis. However, the broad-range caspase inhibitor VAD prevented eosinophil death, indicating that caspases are also involved within the apoptotic machinery of eosinophils. Functional inhibitor studies suggested that caspase 9 is crucial for both caspase 3 and 8 activation, at least in neutrophils. In contrast, spontaneous apoptosis of neutrophils or eosinophils is unlikely to be the consequence of Fas ligand/Fas receptor molecular interactions. CONCLUSION: The data of this study indicate differences in the usage of caspases between neutrophils and eosinophils. PMID- 11729354 TI - Increased levels of IgA antibodies against desmin in children with coeliac disease. AB - BACKGROUND: In addition to IgA anti-endomysial antibodies, IgA anti-smooth muscle antibodies have been observed in a number of patients with coeliac disease (CoD), but only limited data exist on the frequency and antigen specificity of IgA against cytoskeletal proteins in CoD. METHODS: We evaluated the sera of 42 untreated CoD patients, follow-up sera of 26 CoD patients after treatment, and 116 control sera for IgA reactivity to cytoskeletal proteins from the human umbilical cord (HUC) by immunoblotting, and compared the results with anti-tissue transglutaminase IgA (anti-tTG IgA) ELISA and immunofluorescence results. RESULTS: Serum IgA from CoD patients most frequently recognized a 57-kD antigen in HUC cytoskeletal extract, identified as desmin using mass spectrometry and internal peptide sequencing. Increased IgA reactivity to the human desmin band was detected in 22 children with untreated CoD (52.4%), in 4 treated CoD patients (15.4%), and in 12 control subjects (10.3%) (p < 0.01); similar results were obtained with anti-chicken desmin IgA ELISA. Anti-tTG IgA levels (increased in 71.4% of untreated CoD patients) correlated significantly with anti-desmin IgA levels in untreated CoD, and both autoantibody reactivities decreased in response to a gluten-free diet. Pre-adsorption experiments and affinity purification of anti-desmin IgA antibodies further confirmed that desmin is an IgA autoantigen in CoD and is recognized by antibodies which are not cross-reactive with tTG. CONCLUSIONS: Anti-desmin IgA antibodies are frequently occurring if not the predominant cytoskeletal antibodies in children with untreated CoD and could be related to the disease process in the small intestine. PMID- 11729355 TI - The effect of heat-inactivated Helicobacter pylori on the blastogenic response of peripheral blood mononuclear cells of patients with chronic urticaria. AB - BACKGROUND: Helicobacter pylori, the most important etiologic factor of gastritis and peptic ulcer, has recently been associated with several extradigestive diseases. Previous studies reported conflicting results on H. pylori eradication in chronic urticaria, in that some studies showed a benefit, while others found no effect. METHODS: Peripheral blood mononuclear cells of 24 chronic urticaria patients (13 seropositive/11 seronegative for H. pylori) and 18 healthy controls (9 seropositive/9 seronegative) were stimulated with whole heat-inactivated H. pylori (8 x 10(5), 8 x 10(6 )and 8 x 10(7) bacteria/well), phytohemagglutinin (2 microg/ml) and pokeweed mitogen (5 microg/ml). The proliferative response was determined by (3)H-thymidine incorporation. Helicobacter-specific IgG antibody response was determined by ELISA. RESULTS: There were significantly higher proliferative responses to various concentrations of whole heat-inactivated H. pylori antigen in 6- to 7-day cultures of peripheral blood mononuclear cells of chronic urticaria patients compared to healthy controls. We found a tendency to exhibit a higher proliferative response to either Helicobacter antigens or mitogens in seropositive compared to seronegative patients. CONCLUSION: Our results support the hypothesis that there is an increased lymphocyte reactivity in chronic urticaria, perhaps further enhanced by the presence of H. pylori which, therefore, may be involved as a trigger in the pathogenesis of chronic urticaria. PMID- 11729356 TI - Equal allergenic potency of beta-lactam antibiotics produced by chemical or enzymatic manufacturing--mouse IgE test. AB - BACKGROUND: Cephalexin and amoxicillin are semisynthetic beta-lactam antibiotics with a broad spectrum of antibacterial activity against gram-positive and gram negative microorganisms. Both antibiotics are produced by a new 'green' process in which enzyme technology is used to combine the intermediate structure and the side chain in an aqueous medium to yield cephalexin or amoxicillin, thus avoiding the use of several chemical reagents and volatile organic solvents. As a result of the enzyme technology a new residual protein impurity has been identified. To check for the sensitizing capacity of the residual protein, a mouse IgE test was used to detect differences in the production of specific IgE by chemical or enzymatic preparations of the antibiotics. METHODS: Balb/c female mice were immunized intraperitoneally with alum and conjugates of different amoxicillins or cephalexins with ovalbumin (OVA). After 16 days, the amoxicillin mice were injected with one half the original amount of antigen. After 19-23 days, the sera were tested for specific IgE by the passive cutaneous anaphylaxis assay in Sprague-Dawley rats. The greatest dilution of sera which resulted in a positive response was the titer of specific IgE. RESULTS: No significant differences were found between the titers of specific IgE caused by the chemically and enzymatically produced beta-lactam antibiotics, indicating that the antibiotics are equal in allergenicity. CONCLUSIONS: The data show that a residual level of 35 ppm protein did not affect the allergenic potency of these beta-lactam antibiotics as determined by the mouse allergenicity model. PMID- 11729357 TI - Partial agonistic properties of (+/-)-pindolol at atypical beta-adrenoceptors in the guinea pig gastric fundus. AB - (+/-)-Pindolol ([1-(1H-indol-4-yloxy)-3-[(1-methylethyl)- amino]-2-propanol)]) is a partial agonist at atypical beta-adrenoceptors in the guinea pig gastric fundus. (+/-)-Pindolol induced concentration-dependent relaxation in this tissue. However, the relaxant responses of (+/-)-pindolol were not antagonized by a combination of the selective beta(1)-adrenoceptor antagonist atenolol (10(-4) mol/l) and the selective beta(2)-adrenoceptor antagonist butoxamine (10(-4) mol/l). In the presence of both atenolol and butoxamine, the nonselective beta(1) , beta(2)- and beta(3)-adrenoceptor antagonist (+/-)-bupranolol (10(-5)-10(-4) mol/l) caused a concentration-dependent rightward shift of the concentration response curves for (+/-)-pindolol. Schild plot analyses of (+/-)-bupranolol against (+/-)-pindolol gave the pA(2) value of 5.46 +/- 0.03 and Schild slope was not significantly different from unity. Furthermore, (+/-)-pindolol (10(-5) mol/l) weakly but significantly antagonized the relaxant responses to catecholamines ((-)-isoprenaline, (-)-noradrenaline and (-)-adrenaline), a selective beta(3)-adrenoceptor agonist BRL37344 ((R*,R*)-(+/-)-4-[2-[(2-(3 chlorophenyl)-2-hydroxyethyl)amino]propyl]phenoxyacetic acid sodium salt) and a nonconventional partial beta(3)-adrenoceptor agonist (+/-)-CGP12177A ([4-[3-[(1,1 dimethylethyl)amino]-2-hydroxypropoxy]-1,3-dihydro-2H-benzimidazol-2-one] hydrochloride). These results suggest that (+/-)-pindolol acts as a partial agonist at atypical beta-adrenoceptors in the guinea pig gastric fundus. PMID- 11729358 TI - Inhibitory effect of egualen sodium: a new stable derivative of azulene on histamine release from mast cell-like cells in the stomach. AB - We studied the inhibitory effect of egualen sodium (ES) (sodium 3-ethyl-7 isopropyl-1-azulenesulfonate 1/3 hydrate, KT1-32), a new derivative and more stable compound than azulene, on histamine release from the mucosal histaminocytes and elucidated the mechanism for this action. ES prevented the histamine release from isolated mast cell-like cells of the guinea pig stomach induced by A23187 in a dose-dependent fashion. ES dose-dependently inhibited the histamine release from lung pieces of sensitized guinea pigs induced by an antigen-antibody reaction. ES also inhibited histamine release from rat peritoneal mast cells induced by compound 48/80 or antigen-antibody reaction. ES exhibited the membrane stabilizing activity on DPPC liposomes. These findings suggest that ES may prevent histamine release from histaminocytes induced by various stimuli and the stabilizing action of the cell membrane may be responsible for the inhibition of histamine release. PMID- 11729359 TI - Molecular and pharmacokinetic evaluation of rat hepatic and gastrointestinal cytochrome p450 induction by tamoxifen. AB - Tamoxifen (TAM) is a first-line endocrine treatment for all stages of postmenopausal breast cancer. The cytochrome P450 (CYP) enzymes catalyze the majority of TAM's primary metabolism, producing N-desmethyltamoxifen (DMT) and 4 hydroxytamoxifen (4-OH-TAM) in both humans and rats. CYP 3A isoforms are the predominant subfamily involved in the formation of DMT and recent studies have shown that TAM induces hepatic forms of these enzymes. TAM's inductive effect on gastrointestinal CYP 3A has not been previously reported. The current studies investigated TAM's induction of CYP isoforms (3A and 2B) in female rat gastrointestinal and hepatic tissue at the mRNA, protein, and catalytic level. Since previous studies have not addressed whether TAM induction causes changes to the overall pharmacokinetics (PKs), a rat PK model was used to determine if TAM induced its own metabolism, and/or the metabolism of a CYP 3A substrate, midazolam (MDZ). Phenobarbital (PB) and/or dexamethasone (DEX) were used as positive controls for all studies. TAM significantly induced, or caused a trend towards induction of all studied parameters for hepatic CYP 3A and 2B, whereas intestinal CYP 3A and 2B analysis did not show significant induction by TAM at any level. A study evaluating time-dependent alterations in the PK profile of TAM showed no change in apparent oral clearance (Cl(app)) during two weeks of chronic dosing with TAM. However, the Cl(app) for MDZ was shown to trend towards an increase after two weeks of dosing with TAM, in a second PK study. These combined investigations suggest that TAM is an inducer of rat hepatic CYP 3A and 2B isoforms, and this agent has the potential of influencing the PK of coadministered 3A substrates. PMID- 11729360 TI - Increased nitric oxide accounts for decreased basal vascular tone and responsiveness in the resistance vessels of high-cholesterol-fed rabbits. AB - The objective of this study was to determine the effects of hypercholesterolemia on basal vascular tone and vascular responses to pharmacologic agents in hindquarter resistance vessels. Blood pressure and hindquarter blood flow were measured in conscious rabbits fed a high cholesterol diet (1%) for 17 weeks (HC) compared to age-matched rabbits fed a normal diet (control). Basal hindquarter blood flow and vascular conductance were significantly higher in HC than in control rabbits. Administration of a non-selective nitric oxide synthase (NOS) inhibitor, L-NAME (100 mg/kg) decreased basal hindquarter blood flow and vascular conductance in a greater magnitude in HC than in control rabbits, thus, abolished the differences in both the flow and conductance between 2 groups, indicating that increased NO was responsible for reduced basal vascular tone in the HC rabbits. L-NIL (30 mg/kg), a selective inducible NOS (iNOS) inhibitor had no effects on either flow or conductance. This result does not support the involvement of iNOS. In separate experiments, animals were anesthetized and instrumented with an extracorporeal circuit to measure perfusion pressure under constant blood flow to the hindquarter vascular bed. In the HC group, vascular responses to acetylcholine, S-nitroso-N-acetyl-penicillamine and phenylephrine were all attenuated when compared to the responses in the control rabbits. These results indicate that local overproduction of NO due to hypercholesteremia could desensitize smooth muscle reactivity, thus causing general vascular hyporesponsiveness to vasoactive agents. PMID- 11729361 TI - Effects of diacerein on indomethacin-induced gastric ulceration. AB - We investigated the effect of diacerein, an antiosteoarthritic agent, and its metabolite, rhein, on the production of reactive oxygen species (ROS) from neutrophils as well as the protective effect of diacerein on indomethacin-induced gastric ulceration and its protection mechanism. Rhein inhibited the ROS production from N-formyl-methionyl-leucyl-phenylalanine or phorbol-12-myristate 13-acetate-activated human peripheral neutrophils. Indomethacin-induced gastric ulceration was significantly inhibited by oxygen radical scavengers and 16,16 dimethyl-prostaglandin E(2) (dmPGE(2)) but not by allopurinol. Diacerein inhibited indomethacin-induced gastric ulceration in a dose-dependent manner. Diacerein did not affect the gastric mucosal PGE(2) content. In addition, diacerein inhibited HCl + ethanol-induced gastric ulceration. These data indicate that the inhibitory effect of diacerein on indomethacin-induced gastric ulceration could be mediated not by the augmentation of gastric mucosal PGE(2) production but by the suppression of ROS production based on its inhibition of neutrophil activation. PMID- 11729362 TI - Ketoprofen-induced cyclooxygenase inhibition in renal medulla and platelets of rats treated with caffeine. AB - It has been suggested that caffeine can augment analgesic activity and aggravate side effects of nonsteroidal anti-inflammatory drugs (NSAIDs). The aim of the present study was to investigate a possible interaction between ketoprofen and caffeine on prostaglandin (PG) biosynthesis and cyclooxygenase (COX) mRNA expression in the rat renal medulla ex vivo. Treatment of rats with ketoprofen (60 min before) resulted in a dose-dependent (estimated ID(50) 0.3 mg/kg p.o.) reduction of PGE(2) biosynthesis in renal medulla ex vivo. Ketoprofen (0.3 mg/kg) induced inhibition of PGE(2) biosynthesis was stable between 30 and 180 min and still detectable 300 min after drug administration. Caffeine (10 mg/kg) did not cause a detectable effect on its own, nor did it significantly affect ketoprofen induced inhibition of renal medullary PGE(2) biosynthesis. Similar results were obtained with repeated daily drug administration for 1 week: there was no significant effect of caffeine on ketoprofen-induced inhibition of renal medullary PGE(2) biosynthesis. The absence of significant caffeine effects on ketoprofen-induced inhibition of renal medullary PGE(2) biosynthesis was paralleled by experiments showing no significant effect of caffeine on ketoprofen induced inhibition of platelet thromboxane (TX)B(2) biosynthesis. Additional experiments showed increased COX-2 mRNA expression in the renal medulla 60 min after ketoprofen administration, that was not significantly influenced by concomitant caffeine treatment. Treatment of rats with ketoprofen for 1 week had no significant effects on COX-2 mRNA expression. The present results show that ketoprofen caused inhibition of PGE(2) biosynthesis in the rat renal medulla ex vivo with a potency similar to that reported for in vivo models suggesting that the ex vivo approach is a valid model to test a possible interference of caffeine with ketoprofen-induced COX inhibition. The absence of detectable effects of caffeine on time course or magnitude of ketoprofen-induced suppression of PGE(2) biosynthesis in this model indicates, therefore, that possible adverse actions of co-administered caffeine on renal function are not related to interference with renal COX inhibition. PMID- 11729363 TI - Differential effects of citrate versus PPACK anticoagulation on measured platelet inhibition by abciximab, eptifibatide and tirofiban. AB - BACKGROUND: High levels of glycoprotein (GP) IIb/IIIa receptor inhibition are required to prevent arterial thrombosis following percutaneous coronary intervention. Ex-vivo turbidometric platelet aggregation in citrate anticoagulated blood samples has been the primary method previously utilized to derive dose regimens for administering platelet GP IIb/IIIa inhibitors. Enhanced GP IIb/IIIa binding and inhibition of platelet aggregation for eptifibatide secondary to citrate induced reduction of ionized plasma calcium concentrations has been reported. METHODS/RESULTS: We evaluated the differential effects of citrate versus PPACK anticoagulation on turbidometric platelet inhibition in normal volunteers by eptifibatide, tirofiban or abciximab. The decrease in ionized calcium afforded by citrate was associated with enhanced in vitro platelet inhibition for all three GP IIb/IIIa inhibitors, including abciximab. The magnitude of citrate effect was greatest for eptifibatide. Both tirofiban and abciximab have similar citrate calcium chelation associated enhancement of measured platelet inhibition. CONCLUSION: Accurate assessment and comparison of platelet inhibition by GP IIb/IIIa inhibitors may require avoidance of calcium chelating anticoagulants. PMID- 11729364 TI - Comparison of GP IIB/IIIA inhibitors and their activity as measured by aggregometry, flow cytometry, single platelet counting, and the rapid platelet function analyzer. AB - BACKGROUND: GP IIb/IIIa inhibitors have primarily been used short-term e.g., during PTCA. They failed to show clinical benefit during long-term therapy. One reason might be the absence of a method to monitor inhibitor activity. This study compared platelet aggregometry, the rapid platelet function analyzer (RPFA) test, single platelet counting, and flow cytometric determination of receptor occupancy to measure GP IIb/IIIa-receptor inhibitor activity. METHODS: Increasing doses of abciximab, tirofiban, and eptifibatide were added to whole blood in vitro. Whole blood was used for the RPFA, for single platelet counting and flow cytometry. Platelet rich plasma was prepared for aggregometry. RESULTS: The correlation between aggregometry and RPFA results was linear for abciximab and eptifibatide. Tirofiban was a stronger inhibitor with the RPFA (IC(50) 7.7nM) than with aggregometry (IC(50) 19.6nM). The single platelet counting technique showed that even supratherapeutic concentrations of all three inhibitors could not completely suppress microaggregation. Abciximab concentrations that were equipotent to tirofiban with aggregometry were less potent with regards to the inhibition of microaggregation. This difference was more pronounced with TRAP induced microaggregation than with ADP. The flow cytometric receptor occupancy test showed that occupancy was 95% with 5 microg/ml abciximab and almost 97% with 10 microg/ml. Tirofiban reached a maximum receptor occupancy of 56%, eptifibatide 64%. CONCLUSIONS: While aggregometry is time consuming the RPFA provides results fast and with little variability. There is still a discrepancy between aggregometry and RPFA results for tirofiban. The single platelet counting technique detects the inhibition of microaggregation the relevance of which for the clinical outcome is not known. The flow cytometric receptor occupancy assay is best suited for abciximab. PMID- 11729365 TI - A synopsis of the clinical uses of argatroban. AB - Argatroban, a direct thrombin inhibitor, has been used in Japan since the early 1980's and was recently approved for use in the United States for patients with heparin-induced thrombocytopenia. However, its use has been studied in other clinical settings including, myocardial infarction, percutaneous coronary intervention and cerebral thrombosis. The doses used in the different clinical situations vary, but argatroban offers the advantage of not requiring renal adjustment. Because of its small molecular weight, argatroban has the ability to inhibit both clot bound and soluble thrombin. This paper provides a comprehensive review of both indicated and off label uses of argatroban. Pharmacology, pharmacokinetics, adverse events and drug interactions with argatroban are also discussed. PMID- 11729366 TI - Protein and angiogenic dose-response expression of phVEGF-A(165) gene in rat myocardium. AB - Therapeutic myocardial angiogenesis by means of transient overexpression of angiogenic growth factors is a potential treatment modality for severe ischemic heart disease. This study was undertaken in the rat to examine effects of phVEGF A(165) myocardial transfection in terms of dose-response as regards the number of hVEGF-A expressing cells on one hand and on the other angiogenesis. Non-surgical echocardiography-guided intramyocardial injection of phVEGF-A(165) was done into normoxic or hypoxic (10% O(2)) rats. Cardiomyocytes expressing VEGF-A protein, capillary morphology and density were determined after 5 days. VEGF protein expression was seen in rat cardiomyocytes located around the tip of the injection scar and increased dose-dependently (p<0.05). Microvessel density also increased dose-dependently with phVEGF(165) (p<0.05) and with hypoxia (p<0.05). No vascular tumours were observed. In conclusion, direct intramyocardial injection of phVEGF A(165) in the rat results in a dose-dependent increase both in transfected hVEGF A protein producing cells and in angiogenesis. PMID- 11729367 TI - Bedside coagulometry during intravenous heparin therapy after coronary angioplasty. AB - In order to assess the applicability of a bedside coagulometer for measurement of b-APTT, serial blood samples were obtained from 20 patients receiving intravenous heparin treatment following PTCA, and from 5 healthy volunteers. B-APTT was analysed bedside on the Hemochron coagulometer; p-APTT and p-heparin, measured as factor anti-Xa activity, were analysed ex-vivo in the laboratory. B-APTT values, determined by the Hemochron coagulometer, were closely correlated to p-heparin (r=0.83, p<0.001, SD=52 seconds (sec), n=89), and duplicate determinations of b APTT on the Hemochron coagulometer showed an acceptable repeatability. However, an APTT ratio of 1.5-2.5 was not related to a therapeutic p-heparin level, neither as measured by the Hemochron device nor in the laboratory. BACKGROUND: When administering intravenous heparin during angioplasty procedures, a quick and reliable method for safe and effective monitoring of anticoagulation is necessary. OBJECTIVE: To assess the applicability of a bedside coagulometer, measuring the activated partial thromboplastin time (APTT) in patients receiving intravenous heparin treatment after percutaneous transluminal coronary angioplasty (PTCA). METHODS: In patients with stable angina pectoris, receiving intravenous heparin treatment following PTCA, serial blood samples were obtained by venipuncture and from the arterial sheath for analysis of whole blood APTT (b APTT), and plasma heparin concentration (p-heparin). Additionally, in healthy volunteers blood samples were obtained after a single bolus injection of heparin. B-APTT was analysed bedside on the Hemochron coagulometer; p-APTT and p-heparin, measured as factor anti-Xa activity, were analysed ex-vivo in the laboratory using conventional analytical methods. RESULTS: In 20 patients a total of 94 venous and 69 arterial blood samples were analysed, and in five healthy volunteers analyses were performed in 20 venous blood samples. B-APTT values, determined by the Hemochron coagulometer, were closely correlated to p-heparin (r=0.83, p<0.001, SD=52 seconds (sec), n=89). An APTT ratio of 1.5-2.5 was not related to a therapeutic p-heparin level, however, neither when using APTT assessed by the Hemochron device nor APTT measured in the laboratory. Duplicate determinations of b-APTT on the Hemochron coagulometer showed an acceptable repeatability; the mean difference between duplicate measurements was 4 sec (coefficient of variation (c.v.)=6%, p<0.05, n=163). CONCLUSIONS: In patients receiving intravenous heparin after PTCA treatment, b-APTT values measured by the Hemochron method showed an acceptable repeatability and were significantly correlated to p-heparin. PMID- 11729368 TI - Ecarin clotting time but not aPTT correlates with PEG-hirudin plasma activity. AB - BACKGROUND: Novel antithrombotic agents such as hirudin have shown promise in the therapy of acute coronary syndromes. PEG-hirudin (polyethyleneglycol conjugated hirudin) has been developed to provide a longer plasma half-life and more stable antithrombotic plasma levels. Privious trials indicated a narrow therapeutic window for hirudin and a number of aPTT (activated partial thromboplastin time) monitored trials investigating hirudin in acute coronary syndromes had to be stopped because of intracranial bleeding complications. OBJECTIVES: The present study evaluates the ecarin clotting time (ECT), a parameter based on the conversion of prothrombin by the snake venom enzyme ecarin, for the monitoring of PEG-hirudin therapy. METHODS: Plasma from either healthy volunteers (n=20) or from patients (n=10) suffering from unstable angina pectoris (UAP) was spiked with increasing PEG-hirudin concentrations. In a prospective randomized clinical trial patients with UAP were treated with intravenous PEG-hirudin or heparin over 72 hours. Patients were randomized to the following treatment groups: (1) heparin control group, n=15; (2) PEG-hirudin low dose (0.1 mg/kg bolus, 0.01 mg/kg/h infusion), n=19; (3) intermediate dose (0.15 mg/kg and 0.015 mg/kg/h), n=17; 4) high-dose (0.2 mg/kg and 0.02 mg/kg/h), n=16. Spiked plasma samples and plasma from UAP patients treated with i.v. PEG-hirudin were analyzed for aPTT, ECT, and PEG-hirudin levels. RESULTS: A linear correlation up to the highest therapeutic concentrations could be observed between PEG-hirudin plasma concentrations and the ECT. This was true for both plasma samples spiked with PEG-hirudin in vitro as well as for samples taken from patients treated with i.v. PEG-hirudin (correlation coefficient 0.9, respect.) In contrast the aPTT did not show a reliable linear correlation to PEG-hirudin concentrations. CONCLUSION: Monitoring of PEG-hirudin therapy by ECT may help to avoid inadequate anticoagulation or overdosing. Thus, the safety and efficacy profile of PEG-hirudin therapy is likely to be enhanced by ECT monitoring. PMID- 11729369 TI - Platelet reactivity in coronary ostial blood: a reflection of the thrombotic state accompanying plaque rupture and of the adequacy of anti-thrombotic therapy. AB - BACKGROUND: Optimal anti-thrombotic therapy for acute coronary syndromes (ACS) should suppress pro-thrombotic activity at the site of plaque rupture. We sought to determine whether platelet reactivity is increased in blood in the immediate vicinity of a ruptured plaque and is apparent even when blood is obtained by sampling from a catheter placed proximal to the lesion. METHODS: Blood was obtained from a catheter placed in the aorta and from the same catheter after engaging the culprit coronary artery. Platelet reactivity was determined with the use of flow cytometry by surface expression of P-selectin. RESULTS: In preliminary studies we demonstrated that a marker of thrombin activity, fibrinopeptide A, was similarly increased in blood taken from the coronary sinus and coronary arterial ostium of patients with ACS. Subsequently blood was obtained from the aorta and coronary arterial ostium through a coronary guide catheter for assessment of platelet reactivity in 23 subjects with ACS and 22 subjects with stable angina. The percentage of platelets expressing P-selectin in response to 0.2 microM adenosine diphosphate (ADP) was greater in coronary arterial samples from patients with ACS (aorta=6.1+/-1%, coronary artery=8.8+/ 1.6%, p=0.02) compared with that in patients with stable symptoms (aorta=6.9+/ 1.2, coronary artery=6.5+/-1.4, p=NS). CONCLUSIONS: Coronary arterial blood obtained from the ostium through a coronary guide catheter can be used to determine whether thrombin activity and platelet reactivity are increased in the immediate vicinity of a ruptured atherosclerotic plaque. The simplicity of the approach developed should facilitate its use in future studies designed to determine the impact of optimal suppression of platelet reactivity and the pro thrombotic state before coronary interventions on short- and long-term clinical outcomes. PMID- 11729370 TI - Sulfonylureas are not associated with increased mortality in diabetics treated with thrombolysis for acute myocardial infarction. AB - BACKGROUND: Sulfonylurea compounds may impair ischemic preconditioning and endogenous fibrinolysis. Increased mortality has been reported in diabetics receiving these drugs prior to admission for acute myocardial infarction when treated by direct angioplasty. Although thrombolytics are currently employed far more frequently than direct angioplasty the effect of sulfonylureas on mortality in the setting of thrombolysis has not been previously addressed. METHODS: Two hundred forty five diabetics treated with either accelerated t-PA or streptokinase in a national, multi-center, randomized comparison of argatroban vs. heparin (n=1200) were grouped by anti-diabetic treatment prior to hospitalization, and their outcomes were compared by retrospective analysis. RESULTS: Baseline characteristics were similar in all groups (sulfonylureas: n=121, oral medications other than sulfonylureas: n=17, insulin: n=28, diet alone: n=79). Sulfonylurea use was not associated with increased mortality or adverse event rates. By logistic regression analysis with diet treatment as reference, only prior insulin use was associated with higher risk for mortality at 30 days and 1 year (odds ratios 4.5 and 5.22, respectively, p<0.05). CONCLUSIONS: Sulfonylureas use prior to admission is not associated with adverse outcomes in diabetics treated with thrombolytics for myocardial infarction. Since direct angioplasty may increase mortality in patients taking these drugs, a randomized trial is needed to specifically compare different strategies of acute reperfusion in diabetics.Abbreviated abstract. Increased mortality has been reported in diabetics using sulfonylureas when treated for myocardial infarction by direct angioplasty. No study has specifically addressed the effect of these drugs on outcomes in the setting of thrombolysis. In a retrospective analysis of 245 diabetics treated with thrombolysis in a randomized comparison of argatroban vs. heparin, outcomes were compared in relation to anti-diabetic therapy prior to admission. Sulfonylurea use did not adversely affect prognosis, which was worst among diabetics previously treated with insulin. In conclusion, sulfonylureas do not worsen outcomes of diabetics treated with current thrombolytic regimens in comparison with other anti-diabetic treatments. PMID- 11729371 TI - Concurrent use of reteplase and lepirudin in the treatment of acute anterior wall myocardial infarction. AB - A 61-year-old male with a history of severe heparin-induced thrombocytopenia (HIT) type II after aorto-femoral bypass surgery presented to the emergency department within 8 hours of development of substernal chest pain radiating to the left arm. Electrocardiogram (ECG) on arrival and at 3 hours showed no acute changes; cardiac enzymes revealed minimal MB elevation. Echocardiogram showed normal left ventricular systolic function with mild mitral and tricuspid regurgitation and trace aortic insufficiency. Five hours after arrival, the patient reported a recurrence of severe chest pain. ECG showed marked ST elevations consistent with acute myocardial infarction. Reteplase was administered with concomitant lepirudin. Follow-up ECG showed improvement in ST segment elevation and eventual resolution to pre-event tracing; cardiac enzymes showed slight elevations. Catheterization revealed 90% midstenosis of the left anterior descending artery, which was successfully treated with percutaneous transluminal coronary angioplasty (PTCA) and stent placement. Repeat PTCA was performed 10 days postdischarge due to intraluminal stent occlusion. The patient was doing well at 6 months follow-up. PMID- 11729372 TI - Posttraumatic isolated infarction in the territory of Heubner's and lenticulostriate arteries: case report. AB - A 12 year old male had a secondarily generalized epileptic seizure and a subsequent right hemiparesis with fasciobrachial predominance after a closed head injury. His seizures responded to antiepileptic drug therapy immediately. Computerized tomographic scanning and magnetic resonance imaging showed an acute infarct of the head of the left caudate nucleus, indicating the isolated occlusion of the left recurrent artery of Heubner and lateral lenticulostriate arteries. Pathologies leading to vasculitis and embolism were also looked for, but no finding of associated systemic disease could be disclosed. We present this case since posttraumatic infarction in the territory of the deep perforators such as recurrent artery of Heubner and lateral lenticulostriate arteries are exceptionally rare conditions especially in this age group. PMID- 11729373 TI - For the betterment of the family care for the aged with dementia. AB - To investigate the feeling of burden and needs in care, a questionnaire was made which consists of "feeling of burden in care" (family Maslash Burnout Inventory for care or M.B.I. in short) and "family needs" (own making). It was mailed to ninety-nine caregiver of the family with the aged with dementia who cared them at home utilizing day-care service at a special nursing home for the aged, and attended family class once a month. 67 valid answers were obtained. The average age of caregiver was 57.0 years; seventy-six percent of them were female. Their difficulties were own health condition and lack of a cooperator for care. The main component analysis of family M.B.I. for care showed that fatigue was the main factor, indicating that it is very important to provide the families with mental support. Many caregivers thought that their care were useful and worthy; however score of "emotional exhaustion" were higher and score of "self attainment" were lower. PMID- 11729374 TI - Tamoxifen interacts with NEU/C-ERBB-2 receptor and inhibits growth of human malignant glioma cell lines. AB - The effects of tamoxifen, an antiestrogen, on the inhibition of protein tyrosine phosphorylation in neu/c-erbB-2 receptor, DNA synthesis and proliferation were evaluated using the malignant glioma cell lines U25 IMG and T98G which overexpressing neu/c-erbB-2. Pretreatment of two cell lines with tamoxifen resulted in a dose dependent inhibition of tyrosine phosphorylation as well as DNA synthesis and cell growth in two cell lines correlatively. The results support the hypothesis that activated protein tyrosine kinase receptors are involved in the proliferation of glioma cells. Tamoxifen may be useful in the treatment of malignant glioma. PMID- 11729375 TI - Leptin stimulates rat aortic smooth muscle cell proliferation and migration. AB - Leptin, a peptide secreted from adipose tissue, plays an important role in the regulation of food intake and energy expenditure. In obese patients, plasma leptin levels are elevated and obesity is one of the major risk factors for cardiovascular diseases. Therefore, in this study, we investigated the effect of leptin on vascular smooth muscle cell (VSMC) functions. Cultured rat aortic VSMC expressed 130-kDa short form of leptin receptor. Leptin stimulated both proliferation and migration of VSMC. Leptin stimulated phosphorylation and activation of mitogen-activated protein (MAP) kinases, and also increased phosphatidylinositol (PI) 3-kinase activity. Further, two distinct PI 3-kinase inhibitors, wortmannin and LY294002 inhibited the migratory effect of leptin. These results demonstrate that leptin is a proliferative and migratory factor for VSMC, implying that leptin may play a role in the formation and development of vascular lesions. PMID- 11729376 TI - Presidential Address: Surgical education in the third millennium. PMID- 11729377 TI - Is there still a need for living-related liver transplantation in children? AB - OBJECTIVE: To assess and compare the value of split-liver transplantation (SLT) and living-related liver transplantation (LRT). SUMMARY BACKGROUND DATA: The concept of SLT results from the development of reduced-size transplantation. A further development of SLT, the in situ split technique, is derived from LRT, which itself marks the optimized outcome in terms of postoperative graft function and survival. The combination of SLT and LRT has abolished deaths on the waiting list, thus raising the question whether living donor liver transplantation is still necessary. METHODS: Outcomes and postoperative liver function of 43 primary LRT patients were compared with those of 49 primary SLT patients (14 ex situ, 35 in situ) with known graft weight performed between April 1996 and December 2000. Survival rates were analyzed using the Kaplan-Meier method. RESULTS: After a median follow-up of 35 months, actual patient survival rates were 82% in the SLT group and 88% in the LRT group. Actual graft survival rates were 76% and 81%, respectively. The incidence of primary nonfunction was 12% in the SLT group and 2.3% in the LRT group. Liver function parameters (prothrombin time, factor V, bilirubin clearance) and surgical complication rates did not differ significantly. In the SLT group, mean cold ischemic time was longer than in the LRT group. Serum values of alanine aminotransferase during the first postoperative week were significantly higher in the SLT group. In the LRT group, there were more grafts with signs of fatty degeneration than in the SLT group. CONCLUSIONS: The short- and long-term outcomes after LRT and SLT did not differ significantly. To avoid the risk for the donor in LRT, SLT represents the first line therapy in pediatric liver transplantation in countries where cadaveric organs are available. LRT provides a solution for urgent cases in which a cadaveric graft cannot be found in time or if the choice of the optimal time point for transplantation is vital. PMID- 11729378 TI - Auxiliary partial orthotopic versus standard orthotopic whole liver transplantation for acute liver failure: a reappraisal from a single center by a case-control study. AB - OBJECTIVE: To reappraise the results of auxiliary partial orthotopic liver transplantation (APOLT) compared with those of standard whole-liver transplantation (OLT) in terms of postoperative death and complications, including neurologic sequelae. SUMMARY BACKGROUND DATA: Compared with OLT, APOLT preserves the possibility for the native liver to recover, and to stop immunosuppression. METHODS: In a consecutive series of 49 patients transplanted for fulminant or subfulminant hepatitis, 37 received OLT and 12 received APOLT. APOLT was done when logistics allowed simultaneous performance of graft preparation and the native liver partial hepatectomy to revascularize the graft as soon as possible. Each patient undergoing APOLT (12 patients) was matched to two patients undergoing OLT (24 patients) according to age, grade of coma, etiology, and fulminant or subfulminant type of hepatitis. All grafts in the study population were retrieved from optimal donors. RESULTS: Before surgery, both groups were comparable in all aspects. In-hospital death occurred in 4 of 12 patients undergoing APOLT compared with 6 of 24 patients undergoing OLT. Patients receiving APOLT had 1 +/- 1.3 technical complications compared with 0.3 +/- 0.5 for OLT patients. Bacteriemia was significantly more frequent after APOLT than after OLT. The need for retransplantation was significantly higher in the APOLT patients (3/12 vs. 0/24). Brain death from brain edema or neurologic sequelae was significantly more frequent after APOLT (4/12 vs. 2/24). One-year patient survival was comparable in both groups (66% vs. 66%), and there was a trend toward lower 1-year retransplantation-free survival rates in the APOLT group (39% vs. 66%). Only 2 of 12 (17%) patients had full success with APOLT (i.e., patient survival, liver regeneration, withdrawal of immunosuppression, and graft removal). One of these two patients had neurologic sequelae. CONCLUSIONS: Using optimal grafts, APOLT and OLT have similar patient survival rates. However, the complication rate is higher with APOLT. On an intent-to-treat basis, the efficacy of the APOLT procedure is low. This analysis suggests that the indications for an APOLT procedure should be reconsidered in the light of the risks of technical complications and neurologic sequelae. PMID- 11729379 TI - Improvements in early behavior of rat kidney allografts after treatment of the brain-dead donor. AB - OBJECTIVE: To improve the quality of organs from brain-dead donors by assessing the influence of alternative strategies on the early behavior of kidneys after transplantation into unmodified hosts. SUMMARY BACKGROUND DATA: Kidneys transplanted from living donors perform consistently better than those from cadaver sources. The authors have recently shown that donor brain death produces inflammatory changes in peripheral organs within hours, amplifies coincident ischemia-reperfusion injury, and accelerates acute and chronic rejection. Normalization of the graft by donor hormone treatment has hitherto been unsuccessful. METHODS: A standardized rat model of brain death was used. Experimental groups included recipients of allogeneic grafts from living and brain-dead donors (F344-->LEW). Donors were treated immediately after induction of brain death either with intravenous steroids, which block inflammatory cytokine release, or a soluble P-selectin glycoprotein ligand (sPSGL), which blocks initial selectin-mediated cellular adhesion. Kidney grafts were examined serially up to 10 days by morphology, immmunohistology, and reverse transcriptase polymerase chain reaction. RESULTS: Overall survival of ummodified recipients of kidneys from brain-dead donors was significantly reduced versus living donors. Animals with organs from brain-dead donors that had received steroids or sPSGL survived significantly longer than those from untreated brain-dead donors. The intensity of ischemia-reperfusion injury and of acute rejection was reduced. Cellular infiltration and transcription of mRNA of representative proinflammatory mediators were diminished. CONCLUSIONS: Treatment of organ donors at the time of brain death markedly improves organ quality after kidney transplantation, upgrading it to that from a living donor. PMID- 11729380 TI - Laparoscopic cholecystectomy versus mini-laparotomy cholecystectomy: a prospective, randomized, single-blind study. AB - OBJECTIVE: To analyze outcomes after open small-incision surgery (minilaparotomy) and laparoscopic surgery for gallstone disease in general surgical practice. METHODS: This study was a randomized, single-blind, multicenter trial comparing laparoscopic cholecystectomy (LC) to minilaparotomy cholecystectomy (MC). Both elective and acute patients were eligible for inclusion. All surgeons normally performing cholecystectomy, both trainees under supervision and consultants, operated on randomized patients. LC was a routine procedure at participating hospitals, whereas MC was introduced after a short training period. All nonrandomized cholecystectomies at participating units during the study period were also recorded to analyze the external validity of trial results. The randomization period was from March 1, 1997, to April 30, 1999. RESULTS: Of 1,705 cholecystectomies performed at participating units during the randomization period, 724 entered the trial and 362 patients were randomized to each of the procedures. The groups were well matched for age and sex, but there were fewer acute operations in the LC group than the MC group. In the LC group 264 and in the MC group 150 operations were performed by surgeons who had done more than 25 operations of that type. Median operating times were 100 and 85 minutes for LC and MC, respectively. Median hospital stay was 2 days in each group, but in a nonparametric test it was significantly shorter after LC. Median sick leave and time for return to normal recreational activities were shorter after LC than MC. Intraoperative complications were less frequent in the MC group, but there was no difference in the postoperative complication rate between the groups. There was one serious bile duct injury in each group, but no deaths. CONCLUSIONS: Operating time was longer and convalescence was smoother for LC compared with MC. Further analyses of LC versus MC are necessary regarding surgical training, surgical outcome, and health economy. PMID- 11729381 TI - Impaired quality of life 5 years after bile duct injury during laparoscopic cholecystectomy: a prospective analysis. AB - OBJECTIVE: To assess the impact of bile duct injury (BDI) sustained during laparoscopic cholecystectomy on physical and mental quality of life (QOL). SUMMARY BACKGROUND DATA: The incidence of BDI during laparoscopic cholecystectomy has decreased but remains as high as 1.4%. Data on the long-term outcome of treatment in these patients are scarce, and QOL after BDI is unknown. METHODS: One hundred six consecutive patients (75 women, median age 44 +/- 14 years) were referred between 1990 and 1996 for treatment of BDI sustained during laparoscopic cholecystectomy. Outcome was evaluated according to the type of treatment used (endoscopic or surgical) and the type of injury. Objective outcome (interventions, hospital admissions, laboratory data) was evaluated, a questionnaire was filled out, and a QOL survey was performed (using the SF-36). Risk factors for a worse outcome were calculated. RESULTS: Median follow-up time was 70 months (range 37-110). The objective outcome of endoscopic treatment (n = 69) was excellent (94%). The result of surgical treatment (n = 31) depended on the timing of reconstruction (overall success 84%; in case of delayed hepaticojejunostomy 94%). Five patients underwent interventional radiology with a good outcome. Despite this excellent objective outcome, QOL appeared to be both physically and mentally reduced compared with controls (P <.05) and was not dependent on the type of treatment used or the severity of the injury. The duration of the treatment was independently prognostic for a worse mental QOL. CONCLUSIONS: Despite the excellent functional outcome after repair, the occurrence of a BDI has a great impact on the patient's physical and mental QOL, even at long-term follow-up. PMID- 11729382 TI - Influence of resection margins on survival for patients with pancreatic cancer treated by adjuvant chemoradiation and/or chemotherapy in the ESPAC-1 randomized controlled trial. AB - OBJECTIVE: To assess the influence of resection margins on survival for patients with resected pancreatic cancer treated within the context of the adjuvant European Study Group for Pancreatic Cancer-1 (ESPAC-1) study. SUMMARY BACKGROUND DATA: Pancreatic cancer is associated with a poor long-term survival rate of only 10% to 15% after resection. Patients with positive microscopic resection margins (R1) have a worse survival, but it is not known how they fare in adjuvant studies. METHODS: ESPAC-1, the largest randomized adjuvant study of resectable pancreatic cancer ever performed, set out to look at the roles of chemoradiation and chemotherapy. Randomization was stratified prospectively by resection margin status. RESULTS: Of 541 patients with a median follow-up of 10 months, 101 (19%) had R1 resections. Resection margin status was confirmed as an influential prognostic factor, with a median survival of 10.9 months for R1 versus 16.9 months months for patients with R0 margins. Resection margin status remained an independent factor in a Cox proportional hazards model only in the absence of tumor grade and nodal status. There was a survival benefit for chemotherapy but not chemoradiation, irrespective of R0/R1 status. The median survival was 19.7 months with chemotherapy versus 14.0 months without. For patients with R0 margins, chemotherapy produced longer survival compared with to no chemotherapy. This difference was less apparent for the smaller subgroup of R1 patients, but there was no significant heterogeneity between the R0 and R1 groups. CONCLUSIONS: Resection margin-positive pancreatic tumors represent a biologically more aggressive cancer; these patients benefit from resection and adjuvant chemotherapy but not chemoradiation. The magnitude of benefit for chemotherapy treatment is reduced for patients with R1 margins versus those with R0 margins. Patients with R1 tumors should be included in future trials of adjuvant treatments and randomization and analysis should be stratified by this significant prognostic factor. PMID- 11729384 TI - A new surgical concept for rectal replacement after low anterior resection: the transverse coloplasty pouch. AB - OBJECTIVE: To analyze the feasibility, safety, complication and death rates, and early functional results of the transverse coloplasty pouch procedure after low anterior rectal resection and total mesorectal excision. SUMMARY BACKGROUND DATA: The authors previously developed a novel neorectal reservoir, the transverse coloplasty pouch, in an animal model; they report the first clinical data of a prospective phase 1 study. METHODS: Forty-one patients underwent low anterior rectal resection with total mesorectal excision for rectal cancer (n = 37) or benign pathology (n = 4). The continuity was restored with a transverse coloplasty pouch anastomosis, and the colon was defunctionalized for 3 months. Patients were followed up at 2-month intervals for functional outcome. RESULTS: Intraoperative complications occurred in three patients (7%), none related to the transverse coloplasty pouch. There were no hospital deaths and the total complication rate was 27% (11/41); an anastomotic leakage rate of 7% was recorded. The stool frequency was 3.4 per 24 hours at 2 months follow-up and gradually decreased to 2.1 per 24 hours at 8 months. Stool dysfunctions such as stool urgency, fragmentation, and incontinence grade 1 and 2 were regularly observed until 6 months; the incidence significantly decreased thereafter. None of the patients had difficulties in pouch evacuation. CONCLUSIONS: The transverse coloplasty pouch is a small-volume reservoir that can safely be used for reconstruction after sphincter-preserving rectal resection. The early functional outcome is favorable and can be compared to other colonic reservoirs. The concept of reducing early dysfunction seen after straight coloanal anastomosis and avoiding long-term problems of pouch evacuation is supported by this study. Future trials will compare the transverse coloplasty pouch with other techniques of restorative resections of the rectum. PMID- 11729383 TI - Identification of disease-specific genes in chronic pancreatitis using DNA array technology. AB - OBJECTIVE: To use DNA arrays to analyze the differential gene expression patterns in the normal pancreas and in pancreatic diseases. SUMMARY BACKGROUND DATA: Genome-wide gene expression analysis will provide new insights into gene function and cause of disease. METHODS: RNA was extracted from eight normal pancreatic specimens, eight specimens with chronic pancreatitis (CP), and eight pancreatic cancer (PCa) tissues. Poly A(+) RNA was purified, reverse-transcribed, and converted into cRNA using biotinylated nucleotides. The HuGeneFL DNA array containing 5,600 full-length human genes was used for analysis. RESULTS: First, normal pancreatic tissues were analyzed in comparison with a panel of other normal tissues (colon, liver, prostate, lung, lymph node). This analysis revealed 11 signature genes that were selectively expressed in the pancreas (e.g., pancreatic elastase-IIA). Comparison of the expression of 5,600 genes between the normal pancreas, CP, and PCa specimens showed that the expression of 34 genes was decreased in CP tissues compared with normal pancreatic tissues, and that the expression of all of these genes was simultaneously decreased in PCa. In addition, the expression of 157 genes was increased in CP tissues compared with the normal pancreas. Of those, 152 genes were simultaneously increased in PCa. Thus, only 5 of 5,600 genes were significantly overexpressed in CP compared with both normal pancreas and PCa. CONCLUSIONS: The majority of alterations observed in CP are present in PCa, and the number of genes whose expression is selectively deregulated in CP is surprisingly small. These results may provide new insight into the pathobiology of CP and help identify certain molecular alterations that might serve as targets for new diagnostic tools and disease-specific therapy. PMID- 11729385 TI - One- or two-stage procedure for restorative proctocolectomy: rationale for a surgical strategy in ulcerative colitis. AB - OBJECTIVE: To analyze the results of different strategies for restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) in ulcerative colitis. SUMMARY BACKGROUND DATA: No commonly accepted criteria exist for choosing between the one-stage or the two-stage procedure (with or without temporary diverting ileostomy) for IPAA. The authors analyzed the outcome of patients principally suitable for either of the two alternative surgical strategies. METHODS: A matched-pair control study was performed, comparing surgical details and the early and late outcome of the one-stage (study group, n = 57) versus the two stage procedure (control group, n = 114), for IPAA. RESULTS: No differences were found between the study group and the control group regarding the matching criteria gender, median age at IPAA, systemic corticoid medication, or activity of colitis. Comparing the patients who underwent a one-stage procedure with those who underwent a two-stage procedure, the proportion of patients without complications was significantly higher (P =.0042) and the frequency of late complications was significantly lower (P =.0022) in patients who underwent the one-stage procedure. The percentage of patients with anastomotic strictures was significantly higher in the control group than in the study group (P =.0022). No significant difference was found between the two groups regarding early complications, pouch-related septic complications, pouchitis, median duration of surgery for IPAA, median blood loss, need for transfusion, or median hospital stay. CONCLUSIONS: In patients with ulcerative colitis in whom there is a choice between a one-stage procedure or a two-stage procedure with a defunctioning ileostomy, the one-stage procedure is clearly superior. This finding is of great clinical relevance both for the subjective interests of the patient and from an economic point of view. PMID- 11729386 TI - Analysis of allelic imbalance in patients with colorectal cancer according to stage and presence of synchronous liver metastases. AB - OBJECTIVE: To investigate the relationship between number and location of allelic imbalances (AI) and local tumor progression according to Astler-Coller classification. SUMMARY BACKGROUND DATA: Spontaneous errors in DNA replication (i.e., allelic imbalance or microsatellite instability) have been suggested to play an important role in carcinomatous transformation as reflecting alterations of gene function. METHODS: One hundred two consecutive patients with colorectal carcinoma undergoing surgical resection were included in this study. Patients were distributed according to the Astler-Coller classification as stages A (n = 7), B1 (n = 15), B2 (n = 24), C (n = 31), and D (n = 25). Fluorescent polymerase chain reaction was performed on frozen tumor, normal colon mucosa, and blood DNA at 35 microsatellite markers. Allelic imbalance frequency was compared with tumor staging. RESULTS: The percentage of AI was significantly higher in stage D than in A/B1 and B2. In addition, the percentage of AI was significantly higher in 10 synchronous colorectal liver metastases than in stage A/B1 and B2 tumors. However, the allelotyping revealed a subgroup of A/B1 tumors with a high AI frequency. Statistical analysis showed that the presence of AI at microsatellites D1S305, D2S138, D3S1282, D17S790, and D22S928 presented a significantly positive correlation with stages. CONCLUSION: The frequency of AI significantly correlates with tumor progression of colorectal cancer. Primary tumors with synchronous colorectal liver metastases showed a higher percentage of AI, suggesting that a frequency of AI greater than 35% with this selection of markers indicates a high risk of local progression and of development of metastases. PMID- 11729387 TI - Preoperative diagnostic value of [(18)F] fluorodeoxyglucose positron emission tomography in patients with radioiodine-negative recurrent well-differentiated thyroid carcinoma. AB - OBJECTIVE: To assess the utility of 2-[(18)F] fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) to detect recurrent disease in the follow-up of patients with well-differentiated thyroid cancer (WDTC) who have negative diagnostic (131)I scans and abnormal thyroglobulin levels. SUMMARY BACKGROUND DATA: In general, patients with WDTC have an excellent long-term prognosis when appropriate surgical treatment and follow-up are carried out. After total thyroid ablation, whole-body (131)I scintigraphy and measurement of serum thyroglobulin are useful diagnostic tools to detect persistent or recurrent malignancy. In case of tumor dedifferentiation, decreased or lost iodine-accumulating ability may lead to false-negative (131)I scanning results. The diagnostic and therapeutic delay is responsible for a poor prognosis in this subgroup of patients. Efforts have been made in the search for suitable imaging modalities capable of early detection of recurrent thyroid carcinoma. METHODS: The authors prospectively analyzed 24 patients with WDTC, negative results of whole-body (131)I scintigraphy, and elevated serum thyroglobulin concentrations. Attenuation corrected whole-body FDG-PET scans from the neck to the upper legs were performed. In addition, all patients underwent cervical ultrasonography. The results of the imaging studies were compared with histopathologic findings. If no resection of the suspicious lesion was carried out, computed tomography data were used as control criteria. RESULTS: Overall, FDG-PET disclosed 38 hot spots. The sensitivity of the method was 94.6%, but the specificity was lower (25.0%). The diagnostic accuracy was 87.8%. There were three false-positive results in two patients with benign cervical lymph nodes. In one patient with regional lymph node metastases in the neck, two false-negative results were obtained. Ultrasound classified both findings as malignant, however. Because of unexpected findings, FDG-PET suggested potential modification of the surgical management in nine patients. Distant metastases could be disclosed using FDG-PET in only three patients. CONCLUSIONS: FDG-PET is a useful diagnostic tool in the follow-up of thyroidectomized patients with WDTC, negative (131)I scanning results, and abnormal serum thyroglobulin concentrations. The method detects metastatic disease in 94.6% of cases. PET results changed surgical tactics in a significant number of patients. Accurate staging of locoregional cancer recurrence in the neck may be consummately obtained by concomitant analysis of PET and ultrasound results. PMID- 11729388 TI - [Correlation between Blatella germanica and Periplaneta americana in allergic patients]. AB - BACKGROUND: It has been identified shared allergens from whole body between Blatella germanica and Periplaneta americana. It has been reported high correlation between both cockroaches. OBJECTIVE: To estimate correlation between skin test for two major species of intradomiciliary cockroaches. METHODS: It was included patients with clinical diagnostic of allergy pathology whom attended to external consult of Allergy and Clinical Immunology Service of the Hospital Universitario de Puebla in a period of six months, prick skin tests with standardized antigens of Blatella germanica and Periplaneta americana were made. RESULTS: One hundred twenty patients were enrolled in the study, 54.2% were women and 45.8% were men, with a mean age of 15.7 years old. The correlation coefficient between Germany cockroach and American cockroach to erythema was 0.49 and to papule was 0.40. CONCLUSION: The outcomes showed a moderate correlation or a considerable relation between both cockroaches whether erythema or papule. Based on the results presented, we suggest to evaluate both antigens in order to improve diagnostic accuracy. PMID- 11729389 TI - [Risk factors in asthmatic patients that suffer from systemic arterial hypertension]. AB - OBJECTIVE: To determine the risk factors in asthmatics patients with high blood pressure (HBP). METHODS: This was an observational, retrospective, and comparative clinical study, with 132 subjects, divided in 2 groups, sex and age paired. 66 patients with asthma and HBP, and 66 with HBP only as a control group. RESULTS: We identified 69.69% females, age groups from 37-69 years, average 32. Asthma evolution ranged from 1-50 years average 9.60 and HBP evolution ranged from 1-40 years, average 12.21. Hereditary risks allergy were 34.84% (P = NS), pets indoors 16.55% (P = NS), home carpets 16.66% (P = NS), smoking 42.42% (P = 0.001), feather pillows 10.60% (P = NS), 51.51% patients had different diseases beside asthma and HBP (P = 0.0001). CONCLUSION: Asthma and HBP are chronic diseases that require a surveillance long period. Consequently, it is necessary early risk factors diagnosis to design strategies to do in a positive form, breaking the health-disease chain. Smoking and the presence of other disease factors were significatives, limiting the quality of life of these patients. PMID- 11729390 TI - [Effect of immunotherapy with allergens in asthmatic children with integral treatment]. AB - OBJECTIVE: To analyze the effect of the immunotherapy with allergens (ITA) in asthmatic pediatric patients with integral treatment. MATERIAL AND METHODS: We included children with allergic rhinitis and asthma to treatment in the Clinic of Asthma and Allergy. We made a monthly evaluation of symptoms and annual of laboratory tests. RESULTS: The results are described in averages, G1 followed by G2. They were 16 (12 M/4 F) and 17 (11 M/F) patients with ages of 6.1 +/- 2.8 and 6.2 +/- 2 years. They presented 1.2 +/- 0.5 and 1.3 +/- 0.5 crisis per month and also 4.9 +/- 3.4 and 4.8 +/- 3 hospitalizations per year. In all the treatment began with CGIDS in spray and in 8 and 7 BDP was also used. The use of b-agonists at home was bigger in G1: 3 +/- 2.1 vs 0.6 +/- 0.7 (p = 0.0004). Acute treatment in more occasions was granted in G1: 21 vs 2 (p < 0.0001). It was bigger the number of applications of CGIDS in G1: 30.1 +/- 5.3 vs 15.4 +/- 2.7 (p < 0.0001) and of immunotherapy with allergens 8.1 +/- 9.6 vs 5.1 +/- 6.4 (p = 0.1). CONCLUSIONS: The asthmatic pediatric patients on integral treatment present a bigger and quicker improvement of their rhinitic and asthmatic symptoms when they are also treated with specific IT, even from the beginning of the treatment, they also improve their general immunity and they present reduction of allergic markers. They also use less medications, they present less acute exacerbation's and less hospitalizations. PMID- 11729391 TI - [Nasal allergenic provocation test]. AB - This is a method to evaluate both specific sensitivity to allergens in the nasal mucosa, IgE-mediated hypersensitivity, and antiinflammatory and antiallergic drugs efficacy, whose objectives are for research in diagnosis and treatment. The method is based in allergen extracts delivery in the nasal mucosa and the post challenge measurement of rhinitis symptoms, vasoactive mediators release quantification and nasal obstruction degree evaluated by rhinomanometry. Nasal allergen challenge is a procedure of diagnostic and therapeutic evaluation usefulness, that must be performed in selected patients, in adequate facilities, by experts physicians, with standardised allergen dosages, in an specific nasal area, with objective measurements (rhinomanometry, mediators and secretions of the allergic response) and symptoms scoring that allow get reliable results in patients with allergic rhinitis under study. PMID- 11729392 TI - [Allergic rhinitis. Current symptoms]. AB - BACKGROUND: Whatever disease whose etiology is determined by environment should have a periodic check-up for etiologic agents especially for the group of symptoms that accompanies it's clinical features. The purpose of this research is to check if the classic symptomatology described in Allergic Rhinitis (AR) corresponds to the pre: reality or need some modifications in it's diagnostic interpretation. MATERIAL AND METHOD: For this we evaluated in a period of ninety days 438 patients. RESULTS: Both children and adults 58.22% females and 41.78% males aged between seven months and 67 years), who have AR. The highest level of occurrence by was between 5 and 9 years (23.5%. Symptomatology rhinorrea 91.32%, nasal congestion 87.89%, continuous sneezing 84.93%, nasal itching 81.27%, epistaxis 23.74% and nasal. Dryness 22.15%. CONCLUSION: We have found in AR that the vasomotor phenomena and anatomic-vascular lesions are increasing in the affected people who are born in Mexico City and those who have resided for at least five years in Mexico City and it's vicinity. PMID- 11729393 TI - [Hematopoietic growth factors and their side effects]. PMID- 11729394 TI - [The role of yaws in the serologic screening of treponematoses in blood donors in Martinique]. AB - Using TPHA instead of VDRL for syphilis blood-screening since 1995 showed an important increase of positive blood donors in Martinique. Yaws, another treponema disease, has been present on the island until 1975-1980. Usual tests are unable to identify which type--venereal or non venereal--of treponema is involved. Our study, carried out from January 1995 to May 1999, compares actual serological and epidemiological characteristics of TPHA reactive donors to former studies. In our results, the frequency of reactive TPHA is about 1.04% in blood donations. Donors are carrying serological tracks of a past treponema disease with very low rate of antibodies, sometimes linked to yaws. Among donors aged 18 to 30, prevalence is low and is going to become similar to the rate observed in Continental France. This means that this problem will disappear in new donor generations. We suggest the possibility for them to continue blood donation, if their personal preliminary enquiry fits the admission criteria for blood giving. PMID- 11729395 TI - [Statistical Process Control applied to viral genome screening: experimental approach]. AB - During the National Multicentric Study concerning the introduction of NAT for HCV and HIV-1 viruses in blood donation screening which was supervised by the Medical and Scientific departments of the French Blood Establishment (Etablissement francais du sang--EFS), Transcription-Mediated transcription Amplification (TMA) technology (Chiron/Gen Probe) was experimented in the Molecular Biology Laboratory of Montpellier, EFS Pyrenees-Mediterranee. After a preliminary phase of qualification of the material and training of the technicians, routine screening of homologous blood and apheresis donations using this technology was applied for two months. In order to evaluate the different NAT systems, exhaustive daily operations and data were registered. Among these, the luminescence results expressed as RLU of the positive and negative calibrators and the associated internal controls were analysed using Control Charts, Statistical Process Control methods, which allow us to display rapidly process drift and to anticipate the appearance of incidents. This study demonstrated the interest of these quality control methods, mainly used for industrial purposes, to follow and to increase the quality of any transfusion process. it also showed the difficulties of the post-investigations of uncontrolled sources of variations of a process which was experimental. Such tools are in total accordance with the new version of the ISO 9000 norms which are particularly focused on the use of adapted indicators for processes control, and could be extended to other transfusion activities, such as blood collection and component preparation. PMID- 11729397 TI - [The fundamentals of precaution]. AB - The precautionary principle appeared in the health vocabulary, especially in blood transfusion, at the beginning of the 1990s. It is applied to potential risks in case of scientific doubt and corresponds to an hypothesis of risk that must be completely distinguished from the case of an exceptional residual risk. This principle lies on two innovations: the breach of the link between scientific knowledge and decision, and the creation of a context for a new normative value. Because of their consequences, these innovations should generate a debate between professionals about the caution principle's foundations, its conditions of application, and its judicial drawbacks. This article, mainly dealing with the foundations of caution, will also present the social construction of the precaution, its judicial aspects, as well as the change in the relationship to risks induced by it. PMID- 11729396 TI - [Iron overload and myelodysplastic syndromes]. AB - Transfusion of RBC units, the only current treatment for many myelodysplastic syndromes, and excess intestinal absorption of Fe related to dyserythopoiesis often result in iron overload. This condition is associated with high rates of morbidity and mortality. High-risk patients include those with refractory anemia, sideroblastic anemia, 5q-syndrome, patients with a good prognosis (low or lower intermediate international prognosis score), patients having received over 100 RBC units, and patients under the age of 70. Deferoxamine, while it can prevent iron overload, is a strenuous treatment requiring 8-to-12 hour-overnight subcutaneous injections. When patients comply with the regimen, it efficiently prevents mortality due to iron overload, but must be implemented early in the disorder, usually before transfusing 20 RBC concentrates. A simple way of monitoring iron overload is to measure seric ferritin levels and record the number of RBC concentrates. The chelating treatment should be modulated according to age, MDS type, international prognosis score, number of RBC units received, ferritin levels, and most of all, patient tolerance. The direct subcutaneous approach is currently being evaluated by the French Group for Myelodysplasias for its efficiency to prevent disorders, but seems to be both efficient and well complied with (a national protocol is under way). The recent findings on the proteins implied in iron recycling by macrophages after destruction of RBCs, may in the long term, enable us to manage patients with less burdensome treatments and more effective new oral chelates. PMID- 11729398 TI - Reimbursement strategies in an outpatient-based disease management program for the treatment of deep venous thrombosis in an integrated health care system. PMID- 11729399 TI - Deep venous thrombosis: outpatient therapy with low-molecular-weight heparin. PMID- 11729400 TI - Outpatient treatment of DVT in a 170,000-member HMO. PMID- 11729401 TI - Reimbursement of enoxaparin for outpatient therapy. Proceedings of regional meetings. AB - Standard treatment of deep venous thrombosis (DVT) previously required hospitalization so patients could receive continuous intravenous infusion of unfractionated heparin. However, on Dec. 31, 1998, enoxaparin (enoxaparin sodium) became the first low-molecular-weight heparin (LMWH) to receive an indication from the FDA for outpatient treatment of DVT without pulmonary embolism (PE). The manufacturer of enoxaparin, Rhone-Poulenc Rorer (RPR), states that outpatient use of enoxaparin can help managed care organizations (MCOs) and other health care systems to reduce health care costs by curtailing some of the expenses previously associated with hospitalization to treat uncomplicated DVT. To ascertain the extent to which reimbursement policies within managed care might impede patients' access to enoxaparin, RPR conducted 15 regional meetings during the summer of 1999. During these meetings, medical directors and other personnel associated with MCOs provided anecdotal information suggesting that a dominant method of reimbursing enoxaparin for outpatient therapy has yet to emerge in the United States; and that reimbursement methods appear to be divided equally among the pharmacy benefit, the medical benefit, and a "blended" system combining attributes of the pharmacy and medical benefits. PMID- 11729402 TI - Implementing the new HEDIS hypertension performance measure. AB - There is a problem with blood pressure control in the United States--a problem with significant implications for the health and welfare of the populace. This problem is bigger than managed care, but managed care organizations have both unique opportunities and unique obligations to address it. NCQA has responded to this problem, and to the opportunity for better care implicit in it, by introducing into HEDIS a measure that focuses on hypertension control. This measure will add pressure to health plans to address the problem of hypertension control, but it also will create the opportunity for positive recognition for those plans that succeed. The HEDIS hypertension measure is well grounded in both the science of medicine and the science of measurement. But HEDIS measurement alone will not create change. To effect change will require analysis of the problems that limit the delivery of effective care to patients with hypertension. It will require measurement of the success of the key processes of care upon which effective care depends. And it will require response--rational, focused, and operationally effective. These, in turn, will challenge key managers in health plans. Medical directors will have to influence provider behavior. Pharmacy directors will have to leverage pharmacy resources to support efforts to change provider and enrollee behaviors. And QA directors will have to manage a challenging set of measurement activities, from which plans' efforts to improve will be launched. The next few years will not be easy--demands for improvement increase annually, and resources are every year more scarce. Yet the goal is worth the struggle--to transform an industry that the public perceives to be interested in limiting care into one that the public turns to for assurance that care represents high value. Responding effectively to the HEDIS hypertension measure creates a unique opportunity for managed care--to demonstrate to the public that managed care is leading national efforts to improve quality for 50 million Americans needlessly at risk for heart disease and stroke, and in doing so, to demonstrate its commitment to health maintenance--the very foundation of managed care. PMID- 11729403 TI - Glyburide/metformin HCl clinical overview. PMID- 11729404 TI - Roundtable. Blueprint for conversion of patients on metformin and sulfonylurea to Glucovance. PMID- 11729405 TI - NCQA/HEDIS guidelines for diabetes. PMID- 11729406 TI - A retrospective study of persistence with single-pill combination therapy vs. concurrent two-pill therapy in patients with hypertension. PMID- 11729407 TI - Antiplatelet therapy: a vascular medicine perspective. PMID- 11729408 TI - Antiplatelet therapy: a neurology perspective. PMID- 11729410 TI - Clinical and economic implications of atherosclerosis. PMID- 11729409 TI - Antiplatelet pathways development. Panel discussion. PMID- 11729411 TI - Antiplatelet therapy: a perspective from cardiology. PMID- 11729412 TI - Management of type 2 diabetes: update on new pharmacological options. AB - The value of intensive control of blood glucose levels has been clearly established. Data from the UKPDS demonstrated that improving glycemic control will reduce the risk of microvascular complications of type 2 diabetes, such as diabetic retinopathy, nephropathy, and peripheral neuropathy. Further, the metformin study in overweight patients and the epidemiological analysis of the study both demonstrated a reduction in macrovascular complications and mortality related to improved glycemic control. These findings should enhance awareness among both patients and physicians of the dangers of uncontrolled hyperglycemia and the need for early diagnosis and aggressive treatment for patients with type 2 diabetes. Optimal management of type 2 diabetes most often requires a combination of glucose-lowering medications to achieve glycemic control. Current guidelines for combination therapy advise the use of agents with differing and complementary mechanisms of action in order to maximize therapeutic activity and reduce toxicity. Earlier introduction of combination therapy is increasingly being recommended. The new glyburide/metformin combination medication may facilitate earlier, more appropriate and more effective treatment for patients with type 2 diabetes. PMID- 11729413 TI - Disease management approaches to type 2 diabetes. AB - Diabetes is a serious and costly disorder. The economic impact on the health care delivery system in the United States is extremely large, and the cost to individual affected patients in terms of health status, productivity, and quality of life is immeasurable. Population-based DM initiatives are undergoing extensive testing and application. These strategies have been demonstrated to enhance the care delivered to diabetic patients, improve clinical outcomes and quality of life, and reduce the financial toll of managing the disease. These gains are very valuable to the MCO, which is concerned with providing optimal cost-effective patient care in a systematic fashion. PMID- 11729414 TI - Diabetes: prevalence and economic implications. PMID- 11729416 TI - Comprehensive HEDIS measures for diabetic patients. PMID- 11729417 TI - A retrospective study of persistence with single-pill combination therapy vs. concurrent two-pill therapy in patients with hypertension. AB - CONTEXT: Patients with hypertension often fail to control their blood pressure because they do not comply with pharmacologic therapy. It was hypothesized that a greater percentage of patients receiving a single pill combining an ACE inhibitor and a diuretic would persist with therapy than patients receiving both drugs as separate pills. METHODS: Prescription data were obtained from a large commercial pharmacy benefit manager (PBM). The records of presumably newly diagnosed hypertensive patients for whom lisinopril combined with hydrochlorothiazide in a single pill (lisinopril/HCTZ) was prescribed (n = 1,644) were compared with those of patients for whom lisinopril and a diuretic were prescribed concurrently (n = 624). Likewise, the records of patients for whom enalapril maleate combined with hydrochlorothiazide in a single pill (enalapril/HCTZ) was prescribed (n = 969) were compared with those of patients for whom enalapril maleate and a diuretic were prescribed concurrently (n = 705). Patients were regarded as persisting if they renewed their prescription within three times the number of days supplied by the previous prescription. Patients were followed for one year from the date of the initial prescription. RESULTS: At 12 months, the percentages of patients persisting with lisinopril/HCTZ (68.7 percent) and enalapril/HCTZ (70.0 percent) therapy were 18.8 percent and 21.7 percent greater, respectively, than the percentages of patients persisting with lisinopril plus concurrent diuretic therapy (57.8 percent) or enalapril maleate plus concurrent diuretic therapy (57.5 percent). Statistical significance (p < 0.05) was demonstrated at 6 and 12 months for both comparisons. CONCLUSION: The simplification of a drug regimen by using combination therapy in a single pill for hypertension resulted in significant increases in persistence with prescribed therapy. PMID- 11729418 TI - Medication noncompliance: what is the problem? PMID- 11729419 TI - Clinical and economic implications of atherosclerosis. PMID- 11729420 TI - Antiplatelet therapy: a neurology perspective. PMID- 11729421 TI - Antiplatelet therapy: a perspective from cardiology. PMID- 11729422 TI - Antiplatelet therapy: a vascular medicine perspective. PMID- 11729423 TI - Health care in the information age. PMID- 11729424 TI - Issues that will determine health care's future. PMID- 11729426 TI - Humor, risk, and change. PMID- 11729425 TI - Panel discussion. Solutions to health care issues. PMID- 11729427 TI - Panel discussion. The employer's view of health care. PMID- 11729428 TI - Combination DMARD therapy for rheumatoid arthritis. PMID- 11729429 TI - Economic and quality-of-life impact of rheumatoid arthritis. AB - Although RA is a disabling disease with increased mortality and an unknown cure, early treatment with DMARD therapy improves patient outcomes. The economic and humanistic effects of this disease are substantial, with patients becoming increasingly disabled from RA. It is extremely encouraging to the rheumatology community that leflunomide provides significant and sustained improvements in clinical signs and symptoms, and offers practicing physicians an additional option in the treatment of RA. It improves and maintains physical function and HRQoL. Leflunomide's effects were consistent across three studies over two years of treatment. It has been demonstrated to improve physical function as measured by the HAQ DI, and HRQoL as indicated by the SF-36 data, two instruments held up as the gold standard for the assessment of patient outcomes in RA clinical trials. PMID- 11729430 TI - Discussion. Treatment algorithm: managing rheumatoid arthritis. PMID- 11729431 TI - Pharmaceutical agents for the treatment of rheumatoid arthritis. PMID- 11729432 TI - Antidepressant drug interaction considerations. PMID- 11729433 TI - Using pharmacoeconomic data to compare antidepressant therapies. PMID- 11729436 TI - Depression's ripple effect on health status and costs. PMID- 11729434 TI - The Texas Medication Algorithm Project for major depression. PMID- 11729437 TI - Overview of antidepressant therapy. PMID- 11729438 TI - Pharmaceutical industry and payer partnerships to manage costs and improve quality. PMID- 11729439 TI - Pharmaceutical industry and payer partnerships: legal update. PMID- 11729440 TI - Disease management and formulary compliance programs under the HIPAA privacy rule. PMID- 11729442 TI - Comparative pharmacology of proton pump inhibitors. AB - The PPIs are the most effective therapy to suppress gastric acid secretion. These agents decrease acid secretion by inhibiting parietal cell proton pumps. From chemical and pharmacodynamic points of view, subtle differences that exist among the PPIs may influence clinical activity. PMID- 11729443 TI - Optimizing acid-suppression therapy. AB - Acid-related disorders are caused by an imbalance between acid secretion by the gastric parietal cells and the defensive mechanisms of the gastrointestinal tract to protect against the effects of acid. Therapy for acid-related disorders focuses on the control of acidity. Data collected throughout the last decade have demonstrated that PPIs are the most effective therapy for acid-related disorders: PPIs have proven superior to H2RAs and antacids in numerous studies. Five PPIs are currently available in the United States. While all PPIs exert their effect through the same basic mechanism of action, they do not have the same pharmacologic and clinical properties. All PPIs are effective in healing and maintenance of gastric and duodenal ulcers and GERD. The PPIs differ, however, in their ability to control symptoms rapidly and consistently. Due to its more rapid rate of activation, rabeprazole results in a faster onset of action and faster symptom control than other PPIs. Studies comparing rabeprazole to omeprazole found statistically significant differences in the rapidity of symptom relief in patients with gastric ulcer, duodenal ulcer, and GERD. Rapid symptom relief is important to the majority of patients, as their symptoms have an impact on their quality of life. Rapid symptom relief is also important in an environment where patients self-medicate on demand, depending on daily symptoms. Rabeprazole has also been shown to have a more consistent suppression of acid, including at night. Optimizing therapy with PPIs necessitates consideration not only of healing rates of the different available treatments but also of the rapidity and consistency of acid suppression that translate clinically into symptom relief. PMID- 11729444 TI - Health care in the United States: current and future challenges. AB - The last time the United States experienced rapid increases in health care expenditures in the 1980s, the solution was widespread adoption of managed care by employer-sponsored health plans. To a lesser degree, managed care approaches were adopted for some Medicaid and Medicare enrollees. By the mid-1990s, new Medicare reimbursement policies and the widespread adoption of managed care plans had substantially reduced the growth rate of health care expenditures. All published studies show that HMOs and other managed care plans have delivered health care of equal or better clinical quality to traditional indemnity insurance plans at a lower cost. Yet as the economy strengthened during the latter '90s, concerns about overall health care costs lessened, and the public became less willing to accept restrictions on the enrollee's choice of physician and the physician's treatment choices. Public opinion turned against the concept of managed care as a result of backlash from both physicians and consumers. Government mandates altered the ability of managed care to deliver on the promise of higher quality at a reduced cost. Today, health care costs are again rising. The United States is spending significantly more per capita on health care than are other developed nations, with no demonstrable improvement in health outcomes. Estimates suggest that in the next 30 years, health care costs will again rise at a rate faster than that of the economy. Further, rising costs will reflect the impact of an aging population, with more than 20 percent of individuals 65 years or older by the year 2030. Thus, it appears we are again headed for a crisis. However, the environment has changed: As a result of the experience of the last 10 years, the public is now less willing to accept changes to the health care system, and the concept of managed care has negative connotations. Consequently, the crisis may potentially be even worse than that we experienced a decade ago if we are unable to find ways to control health care expenditures. PMID- 11729445 TI - Evidence-based health care: making health policy and management decisions. AB - The paradigm for health care delivery in the United States continues to evolve. Patients and physicians are beginning to reject guidelines for treatment that are solely based on cost reduction and are not evidence based. Future managed care guidelines will rely on the best external clinical evidence about the value of any given therapeutic intervention combined with individual physician experience and patient choice. Managed care organizations will need to play a greater role in the development, evaluation, and incorporation of clinical trials into their strategies. PMID- 11729446 TI - Impact of acid-related disorders in the United States. AB - Acid-related disorders are common conditions that negatively impact quality of life for a significant number of people nationwide. The pathology of these conditions involves an imbalance between acid secretion by gastric parietal cells and the ability of upper GI tract mucosa to defend against the effects of the acid. Therefore, therapy is targeted at elevating gastric pH. PPIs are used to control the effects of excessive acid secretion. This class of drugs has a unique mechanism of action that inhibits the final pathway to gastric acid secretion- the parietal cell proton pump. PMID- 11729447 TI - Technology, DM team up to secure capitation profits. PMID- 11729448 TI - How Wellspan gets doctors to cooperate using only modest incentive pay. AB - This physician group has found that you don't need to have a lot of money for an effective incentive program. Doctors just want to be sure they pass the test and do the right thing. PMID- 11729449 TI - California MSO takes capitation from niche to mainstream. PMID- 11729450 TI - Plans report big losses after shifting away from capitation. PMID- 11729451 TI - Special capitation treatment sought for physicians with high HIV caseloads. PMID- 11729452 TI - Can capitation help small business offer health insurance? PMID- 11729453 TI - Is it time to get your feet wet with consumer-driven models? PMID- 11729454 TI - Capitation must have a role in new health care system, economist maintains. PMID- 11729455 TI - Waterborne cryptosporidiosis outbreak, North Battleford, Saskatchewan, Spring 2001. PMID- 11729456 TI - Bioterrorism redux. PMID- 11729457 TI - Bioterrorism redux. PMID- 11729458 TI - The biopsychosocial versus the supernatural, and other matters. PMID- 11729459 TI - A new way of making doctors. Distance learning for non-traditional students. PMID- 11729460 TI - Traumatic brain injury. North Carolina's challenge. PMID- 11729461 TI - Traumatic brain injury. State of the state. PMID- 11729462 TI - Shaken baby syndrome. Lethal inflicted traumatic brain injury in young children. PMID- 11729463 TI - Primary care for children with brain injury. AB - Children who have sustained a brain injury should receive specialized, coordinated, family-centered care from their primary care providers. By knowing more about the etiology, assessment, associated conditions, and prevention of pediatric TBI, medical providers can better address the multiple needs of the child and family as they cope with new challenges. The management of pediatric brain injury can be complex and time consuming, but we must all be responsive to the needs of these children and their families to ensure their good health, their welfare, and their happiness. PMID- 11729464 TI - Training school psychologists in traumatic brain injury. The North Carolina model. PMID- 11729465 TI - Recreational therapy can help adult brain injury survivors get back into the community. PMID- 11729466 TI - North Carolina's TBI project ACCESS. Assuring coordinated care, education, and support for survivors of pediatric brain injury. AB - Over the past three years, families, school staff, and healthcare personnel in our state have benefited from an innovative service delivery system designed to demonstrate that we can improve the lives of children with TBI. Project ACCESS has used hospital-based pediatric brain injury Community Transition Coordinators to identify, coordinate care, educate, support, and generally improve the lives of thousands of children with TBI. The project also uncovered a host of barriers that adversely affect the future of many of North Carolina's most vulnerable children. We hope that this article helps delineate the challenges faced by our state's healthcare and educational systems, and contributes to the adoption of a statewide model system of care for children with brain injury. PMID- 11729467 TI - To fall or not to fall. Brain injury in the elderly. PMID- 11729468 TI - Sports related head and brain trauma. PMID- 11729469 TI - What can we do about long-term sequelae of traumatic brain injury? AB - It is wrong to think that the needs of persons with brain injury are met or complete in the first few months--or even the first few years--after injury. Some persons with TBI may recover in the acute and out-patient care setting sufficiently to return to home, school, or job, but there is a large group who can greatly benefit from extended care. Extended care can take the form of day activity programs, or respite care for caregivers and injured persons, or supported employment, or home-based services, or long-term residential care. The Commission on Accreditation of Rehabilitation Facilities has recently established standards of care for "assisted living" to provide guidelines for agencies providing long-term care for persons with various disabilities, including TBI. It is my strong opinion that private and government-based providers must work hand in-hand to develop and maintain quality, long-term living options for persons with TBI. Such a collaboration, if sustained, would almost certainly give persons with brain injury and their families a much better chance of returning to pre injury lifestyle or other preferred options. PMID- 11729470 TI - The costs of traumatic brain injury. PMID- 11729471 TI - [COX-2 inhibitor and colon cancer]. AB - It is suggested that nonsteroidal antiinflammatory inhibitors alter the biology of colorectal carcinogenesis. Cyclooxygenase-2, one of target molecules of these drugs, is reported to be upregulated in cancer tissues. Cultured cells which were derived from intestinal epithelium and programmed to express COX-2 showed several phenotypic changes in favor of carcinogenesis, including resistance to apoptosis and enhancement of cell proliferation, angiogenesis, and invasion. Tumor growth implanted in COX-2 null mice was significantly attenuated, but not in COX-1 null or wild type mice, suggesting that COX-2 in stroma also has an important role in tumor growth. Moreover, PGE2, one of COX-2 metabolites, reversed these antitumor effects, indicating that inhibition of PGE2 production has a pivotal role in tumor suppression. However, NSAIDs show antitumor effects in cancer cells lacking COX-1 or COX-2 expression, and some derivatives lacking the ability to suppress COX activity show antitumor effects. These results suggest that COX independent pathway might be involved in antitumor effects of NSAIDs. For the development of novel and effective therapies, it is required to elucidate mechanisms underlying antitumor effects of NSAIDs. PMID- 11729472 TI - [Preoperative chemotherapy for patients with advanced head and neck cancer]. AB - Cases with head and neck squamous cell carcinoma (HNSCC) have clinically advanced tumors. The curative treatments for advanced HNSCC are radiotherapy and/or surgical resection. However, standard treatment alone is less successful for advanced HNSCC. Accordingly, two modalities using chemotherapy are applied as preoperative treatment for HNSCC. First, multi-drug chemotherapy has been administered as neoadjuvant chemotherapy (NAC). As a result during the past 20 years, NAC without a high complete response (CR) rate has never improved the long term outcome of advanced cases. Therefore, the development of intensive chemotherapy regimen with a high CR rate including taxanes is ongoing. On the other hand, organ preservation modality has been under investigation using combined radiotherapy with NAC regimen with a dose reduction of administered chemotherapeutic drugs or a new chemotherapy regimen including taxanes (concomitant or concurrent chemoradiotherapy). In this strategy, impact chemotherapy with almost the same anti-tumor effect as NAC and with a potential of radiation sensitizer is necessary. PMID- 11729473 TI - [Breast cancer]. AB - Neoadjuvant chemotherapy (NAC) represents a new approach based on sound theoretical, pharmacokinetic, and experimental principles. The purpose of NAC is to improve control of the primary site by downstaging and to improve control of micrometastatic disease. NAC has been standard therapy in the management of locally advanced breast cancer. Patients with earlier stage breast cancer may also benefit from treatment with NAC. Recently some investigators have mentioned that NAC can be used instead of adjuvant chemotherapy and would be most appropriate for patients who wish to preserve their breast but who have tumors too large for breast conserving surgery. In this article, we reviewed the present status of NAC (indication, clinical response, pathologic response, survival, possibility of breast conservation, prognostic/predictive factors, neoadjuvant endocrine therapy) and discussed several unanswered questions on NAC (survival benefit, optimal number of treatment cycles, optimal regimens) and future direction. Combined modality therapy including NAC appears to provide excellent local control, the possibility of breast conservation, and, probably, an increased survival rate, at least for some subsets of patients. Furthermore, through sequential sampling, NAC provides indeed the opportunity to identify molecular mechanisms associated with pathologic response and to study the possibility to guide the choice for induction treatment and patient populations submitted to neoadjuvant chemotherapy. PMID- 11729474 TI - [Induction therapy for non-small cell lung cancer]. AB - Induction therapy for non-small cell lung cancer was reviewed. Surgical therapy remains the treatment of choice for resectable non-small cell lung cancer. However, postoperative survival of the patients with locally advanced NSCLC is far from acceptable. Several phase II and phase III trials have been attempted to define whether surgical resection after induction therapy provides better local control and survival than surgery alone. Most studies have reported high response and resectability rates, and long-term follow up of two randomized trials shows that patients having cisplatin-based induction chemotherapy prior to surgical resection were significantly more likely to have better 5-year survival than patients operated without preoperative treatment. However, the results of the randomized trials are still controversial owing to the relatively small and inhomogeneous population used. To identify the future direction of effort in improving the therapy of NSCLC, more sophisticated randomized prospective trials should be conducted. PMID- 11729475 TI - [Neoadjuvant chemotherapy for advanced epithelial ovarian cancer]. AB - Primary surgical cytoreduction followed by paclitaxel/carboplatin combination chemotherapy currently is the treatment of choice for advanced epithelial ovarian cancer. Aggressive surgery is widely accepted as a valid approach to initial cytoreduction of stage III disease, but suboptimal residual disease following primary surgical resection is one of the most important adverse prognostic factors in these patients. Neoadjuvant chemotherapy has been proposed as an alternative approach to conventional surgery for initial management of bulky ovarian cancer, with the goal of improving surgical quality. General acceptance of neoadjuvant chemotherapy as an alternative to primary surgery for patients who are not ideal surgical candidates remains limited, because equivalent or superior survival has not yet been demonstrated in a prospective randomized study. A large scale, prospective, randomized study is being conducted by the European Organization for Research and Treatment of Cancer (EORTC) Gynecologic Cancer Cooperative Group and Gynecologic Oncology Group (GOG) to compare outcomes (overall and progression-free survival, quality of life, treatment complications) of neoadjuvant chemotherapy/interval debulking surgery versus primary cytoreductive surgery/adjuvant chemotherapy in patients with advanced epithelial ovarian carcinoma. PMID- 11729476 TI - [Neoadjuvant chemotherapy for invasive bladder cancer]. AB - Although radical cystectomy is the gold standard for invasive bladder cancer, about half of the patients develop distant metastases within two years after operation. Objectives of preoperative (neoadjuvant) chemotherapy in muscle invasive bladder cancer are eradication of micrometastasis and subsequent improvement of prognosis. Based on favorable outcome of M-VAC (methotrexate, vinblastine, adriamycin, cisplatin) therapy in advanced cases, several clinical trials have been performed. Unfortunately, results of the randomized prospective studies failed to demonstrate survival benefits. However, response to the preoperative chemotherapy has been found to be an excellent prognostic factor, and preservation of the bladder (complete preservation or partial cystectomy) is possible in selected cases with complete response. Moreover, good local control by neoadjuvant chemotherapy enables cystectomy in selected cases of locally advanced cancer once considered unresectable, and may lead to suppression of intraoperative spread of the tumor cells. PMID- 11729477 TI - [Comparison of management of advanced cancer in various organs]. AB - The management of advanced cancer presents the greatest challenge to physicians involved in oncology. There will usually be a large burden of disease; cure is unlikely; and the needs of the patient in terms of pain control and palliation will also be important over and above the direct treatment of the disease. Different issues will arise depending on the site and pathological type of the cancer. Increasingly over the past few years, treatment protocols and guidelines have been developed for different cancers, but these can only be rough guides rather than definite treatment recommendations. Additionally in most cancers advanced disease offers the opportunity for evaluation of new treatments in Phase II studies and other trials. With the new generation of molecular targeted therapies, such as EGFR inhibitors, striking results are being seen in advanced disease that compare favourably with what has been seen previously. Other agents such as those which attack the tumour vasculature may also have promise in this setting. Palliation is also an important aspect of the management of advanced disease, and pain control in particular is an important component of patient management. In summary, the treatment of advanced disease provides a test bed for new agents, but this need to develop better cancer therapies must be balanced against patient needs for a pain-free and comfortable end to life. PMID- 11729478 TI - [Early phase II clinical trial of amrubicin hydrochloride in patients with malignant lymphoma]. AB - An early phase II multi-center collaborative study of amrubicin hydrochloride, a novel synthetic anthracycline derivative anticancer agent, was conducted for malignant lymphoma at 12 institutions nationwide. A total of 41 patients were enrolled in this study between January 1988 and October 1990. Of these, 36 patients, six patients with Hodgkin's disease (HD) and 30 patients with non Hodgkin's lymphoma (NHL), were eligible for the study. The starting dose of amrubicin hydrochloride was 100 mg/m2 (body surface area) and it was administered once every three weeks, in principle. The efficacy was assessed for 34 patients, excluding two patients: one who has not been followed up adequately and the other violated the dosing schedule (once per week). The overall response rates (CR + PR) were 50.0% (3/6) for HD and 42.9% (12/28) for NHL. Furthermore, a relatively high response rate was noted in 8 (36.4%) of 22 NHL patients who had been treated with other anthracycline derivatives prior to the trial. The safety of amrubicin hydrochloride was assessed for 36 eligible patients. Leukopenia (grade 3 or higher) and thrombocytopenia were noted in 21 patients (58.3%) and 10 patients (27.8%), respectively. Anorexia, nausea/vomiting, fever, alopecia, decrease in hemoglobin and elevations of GOT and GPT levels were observed with a relatively high frequency. Other than myelosuppression, the following adverse reactions (grade 3 or higher) occurred during the course of the trial: diarrhea (two patients), alopecia (two patients), stomatitis (one patient), anorexia (one patient), nausea/vomiting (one patient) and fever (one patient). In conclusion, these results indicate that amrubicin hydrochloride is effective in the treatment of patients with malignant lymphoma. PMID- 11729479 TI - [Late phase II clinical study of amrubicin hydrochloride, a novel synthetic anthracycline derivative anticancer agent, for malignant lymphoma]. AB - A late phase II clinical trial of amrubicin hydrochloride, a novel synthetic anthracycline derivative anticancer agent, was conducted at 14 institutions nationwide, in patients with non-Hodgkin's lymphoma. In this multi-center collaborative study, doxorubicin hydrochloride was replaced by amrubicin hydrochloride in CHOP therapy, a standard regimen for non-Hodgkin's lymphoma consisting of cyclophosphamide, doxorubicin hydrochloride, vincristine sulfate and prednisolone. A total of 39 patients were enrolled in this study between January 1996 and March 1998. Among them, 37 patients were eligible for this study. The study drugs were administered to patients with non-Hodgkin's lymphoma according to the following schedule: amrubicin hydrochloride (100 mg/m2, body surface area), cyclophosphamide (750 mg/m2) and vincristine sulfate (1.4 mg/m2, a maximal dose of 2.0 mg/body) were administered intravenously on day one, while prednisolone (60 mg/m2/day) was administered orally on days 1 to 5. This cycle of treatment was repeated every three weeks in principle. The efficacy and safety were assessed for 37 eligible patients. The combined rate for CR + CRu was 70.3% (26/37) and the overall response rate (CR + CRu + PR) was 86.5% (32/37). demonstrating that amrubicin hydrochloride was very effective in the treatment of non-Hodgkin's lymphoma. The most frequent adverse reactions that occurred during the study were myelosuppressions: leukopenia and neutropenia, 100% (37/37); and decreases in hemoglobin levels, 81.1% (30/37). Thrombocytopenia, elevations of serum GOT and GPT levels, anorexia, nausea/vomitting, fever, stomatitis and alopecia were also observed. Although leukopenia and neutropenia of grade 3 or higher were noted in 89.2% (33/37) and 94.6% (35/37), respectively, they were controllable by administrations of G-CSF or solely by follow-up observations. One patient developed intestinal paralysis (grade 4) and another developed hematemesis. In conclusion, these results indicate that amrubicin hydrochloride is an effective agent as a component of combination chemotherapy for non Hodgkin's lymphoma. PMID- 11729480 TI - [Dihydropyrimidine dehydrogenase activity in urothelial cancer--influence of UFT administration on DPD activity]. AB - We measured the dihydropyrimidine dehydrogenase (DPD) activity considered to regulate the sensitivity of pyrimidine fluoride anti-cancer drugs in 50 cases of urothelial cancer (preoperative therapy group 10 cases, non-therapy group 40 cases). The association of DPD activity with pathological factors and TS activity as well as the influence of UFT oral administration on the DPD activity in tumors and blood were studied. The DPD activity in cancer tissue tended to be higher than that in the mucous membranes of the non-tumor region, but the difference was not significant. The DPD activity was high in infiltrative cancer (p < 0.05) but was not associated with atypia. The TS activity and DPD activity were manifested independently. The DPD activity in cancer tissue was significantly higher after UFT administration, with levels of 76.7 +/- 71.0 pmol/mg/min before administration and 220.6 +/- 129.1 pmol/mg/min after (p < 0.05). The DPD activity in peripheral blood was elevated after administration, but no significant difference was found. Since DPD activity is found in urothelial cancer tissue, 5 FU stable to DPD should preferably be used. It is also suggested that using a more potent DPD inhibitor in combination will produce higher anti-tumor effects. PMID- 11729481 TI - [Basic study on the sensitivity test for inhibiting fibroblast proliferation (II)]. AB - The results of in vitro tests against solid tumors have been generally influenced by the mixing in of fibroblasts, and so have SDI tests. We demonstrated that the influence of fibroblasts can be excluded by performing the SDI test on an agar layer, thus significantly inhibiting the growth of fibroblasts in the liquid top layer. In the agar SDI test, SDS is used to dissolve formazan; however, this lowers the sensitivity to some extent. We added additional DMSO to SDS and to raise the sensitivity. PMID- 11729482 TI - [Assessment of intraperitoneal drug distribution during intraperitoneal chemotherapy using saline-injected MR imaging through an implanted catheter-port system]. AB - Intraperitoneal saline-injected MR imaging through an implanted catheter-port system (saline-MRI) was conducted in 7 patients with ovarian tumor after surgical removal of the primary tumor. Two types of T2 weighted coronal images of the abdomen were obtained after saline injection through the implanted catheter-port system. One uses long TE (about 1000 msec) with fat-saturation and thick slices (100 mm thickness) to depict the injected saline alone. The other uses medium TE (about 100 msec) without fat-saturation and thin slices (10 mm thickness) to depict both intraperitoneal saline and abdominal structures. Saline sequentially fills the Douglas pouch, paracolic gutter, Morison's pouch and subphrenic space in most patients. The relation between injected saline and abdominal structures was seen well on T2-weighted images using medium TE. Adhesions of the peritoneum were well demonstrated. In one patient, a catheter perforation to the bowel loop was diagnosed, because the small bowel loop was immediately filled with injected saline. Saline-MRI can be used to depict intraperitoneal drug distribution during intraperitoneal chemotherapy and can diagnose complications related to intraperitoneal chemotherapy. PMID- 11729483 TI - [Hepatic infusion of docetaxel using PEIT for a patient with stage IV breast cancer]. AB - Hepatic infusion of docetaxel using PEIT was performed for a patient with stage IV breast cancer. Docetaxel was effective to a solitary liver metastatic lesion. A 64-year-old woman was admitted to our hospital because of a left breast mass that was bleeding. She was diagnosed with stage IV breast cancer. Surgery was performed on February 16th. The pathological diagnosis was invasive ductal carcinoma, and hormone receptors were negative. Two weeks after operation, monthly docetaxel injections were given together with doxifluidine 400 mg/day p.o., cyclophosphamide 50 mg/day p.o., and fadrozole hydrochloride hydrate 2 mg/day p.o. After two courses, hepatic infusion of docetaxel was performed using PEIT after informed consent. The patient's high serum CEA and CA15-3 level returned to the normal range. A metastatic lesion on CT changed to a cystic pattern. These results suggest that PEIT is worth trying in patients with solitary liver metastasis from breast cancer. PMID- 11729484 TI - [A case of breast cancer with multiple metastases effectively treated with long term administration of docetaxel, aclarubicin and UFT]. AB - A 46-year-old woman had undergone a mastectomy in 1994, and metastases to the lung, bones and liver were detected in 1998. Brain metastases were detected later. Chemotherapy consisting of docetaxel, aclarubicin and UFT was administered, and image diagnosis revealed that CR was achieved. The chemotherapy was continued on a long-term basis, both as an inpatient and outpatient. The total administered doses have reached 2,000 mg of docetaxel and 1,000 mg of aclarubicin. The CR is being maintained as of this writing, 2 years and 4 months after the detection of the metastases. The patient's course continues to be monitored. PMID- 11729486 TI - [A case of postoperative gastric cancer responding to adjuvant chemotherapy with TS-1--a novel oral formation of 5-fluorouracil]. AB - TS-1 is a novel oral formation of 5-fluorouracil, developed using tegafur and two biochemical modulations. We report the case of a patient with multiple lymph node metastases from gastric cancer that markedly responded to TS-1. A 70-year-old man suffering from hematemesis was admitted to our hospital. Computed tomography showed advanced gastric cancer with huge lymph node metastases. After non curative operation, he received adjuvant chemotherapy with TS-1 for 12 weeks. CT revealed that almost complete reduction of the metastatic nodes was obtained. No serious adverse reactions were observed. PMID- 11729487 TI - [A complete response persisting for twelve months with the use of TS-1 in a patient with paraaortic lymph node metastasis of gastric cancer]. AB - A 74-year-old female patient underwent total gastrectomy, splenectomy and D2 lymph node dissection for gastric cancer with non-dissectible paraaortic lymph node metastasis. Pathological examination revealed a high level of metastasis of dissected lymph nodes. The patient received daily oral administration of 100 mg TS-1, a novel oral anticancer agent. Each treatment course consisted of a four week administration followed by two drug-free weeks. A partial response was obtained after the second course and a complete response was observed in the middle of the fourth and after the sixth course. The treatment was stopped because of grade 2 anemia in the middle of the seventh course, but no other adverse effect was observed. Complete response of the treatment persisted for twelve months and the patient has now been in good health without a recurrence for twenty months after surgery. Although the prognosis of gastric cancer with a high level of lymph node metastasis is poor, TS-1 therapy may have a potent efficacy in gastric cancer patients with a high level of lymph node metastasis such as the current case. PMID- 11729485 TI - [Two cases of effective weekly paclitaxel administration for metastatic breast cancer]. AB - We report two cases of effective weekly paclitaxel (TXL) administration for metastatic breast cancer. TXL (80 mg/m2) was infused over 1 hour after short premedication on an outpatient basis. Administration was continued for 3 weeks followed by 1 week rest. Case 1: A 66-year-old woman was treated with adriamycin, cyclophosphamide, doxifluridine (5'-DFUR), fadrozole, medroxyprogesterone acetate (MPA) and pamidronate for pleural, mediastinal and supraclavicular lymph nodes and skull metastases. She complained of hoarseness and supraclavicular and mediastinal lymph nodes had developed, for which she received weekly TXL administration. Metastatic lymph nodes had disappeared 3 months after administration. Case 2: A 61-year-old woman was treated with CMF therapy, 5' DFUR, tamoxifen, MPA fadrozole and pamidronate for bone metastasis and local recurrence. She received weekly TXL administration for an axillary recurrence and contralateral supraclavicular lymph node metastases. Metastatic and recurrent lesions had disappeared 3 months after administration. The toxic events were leukopenia (grade 1) and peripheral neuropathy (grade 1). No major adverse effects were observed in either case. PMID- 11729488 TI - [A case of lung metastasis from colon cancer treated successfully with combined chemotherapy of CPT-11 and 5'-DFUR]. AB - The patient was a 52-year-old woman who had sigmoid colon cancer with liver metastasis and multiple lung metastases. Resection of curability B was performed, and alternating adjuvant chemotherapy consisting of hepatic artery injection of 5 FU and systemic administration of CPT-11 was performed. Lung recurrence was found and no antitumor effect of chemotherapy was observed, so the CPT-11 which had been administered every other week was given every week in a dose of 60 mg/body, half of the original dose. Moreover, 5'-DFUR was administered in a dose of 800 mg/day every day. As a result, lung metastasis tumors were reduced markedly. Adverse events such as nausea, vomiting and depilation were mitigated, and no other toxicity was observed. The patient could thus be treated extremely safely in the outpatient clinic. This was considered to be a valuable case suggestive of the significance of combination chemotherapy of CPT-11 and 5'-DFUR and the importance of appropriate administration of CPT-11. PMID- 11729489 TI - [A case of ovarian cancer with metastatic tumor in the liver with paclitaxel and carboplatin systemic chemotherapy was very effective]. AB - An 82-year-old woman was admitted to the Dept. of Obstetrics and Gynecology, Yamanashi Medical University to try to identify the origin of a liver metastatic tumor. CT examination revealed a small tumor located adjacent to the uterine cervix in a cul-de-sac. With biopsy using MR, it was clearly shown histologically that the origin of the tumor was the ovary. Systemic chemotherapy with paclitaxel and carboplatin was selected as the most reasonable treatment for this case because of the patient's age. After 6 courses of this chemotherapy, the tumor in the cul-de-sac disappeared and the tumor in the liver decreased markedly. Furthermore, no severe side effects were seen during this treatment. This result indicated that systemic chemotherapy with paclitaxel and carboplatin is effective and safe in cases of advanced ovarian cancer. PMID- 11729490 TI - [A case of squamous cell carcinoma of the oral floor showing a complete response to oral administration of UFT]. AB - A 73-year-old male with squamous cell carcinoma of the oral floor (T1N0M0), who had not consented to radical treatment was treated with UFT by oral administration alone. After commencement of the administration, tumor remission was observed at 2 weeks and disappeared clinically at 8 weeks. The total period of oral administration of UFT was twenty-three months, and the total dose was 186.5 g as tegafur. Major adverse effects were pigmentation of hands and liver dysfunction. He remains under observation, and there is no evidence of tumor recurrence. PMID- 11729492 TI - The risk manager's role in creating an organizational patient safety strategy. PMID- 11729491 TI - [Urinary tumor marker for urothelial cancer]. AB - The urinary tumor markers BTA, BFP and NMP22 used for urothelial cancer in Japan are reviewed briefly. We also evaluate and compare the sensitivity and specificity of BTA, BFP and NMP22 with urine cytology in detecting bladder cancer in 24 of our patients. The results showed that the sensitivity with urine cytology, BTA, BFP and NMP22 was 37, 54, 66 and 62% respectively. The specificity of BTA, BFP and NMP22 with urine cytology was 100, 65, 60 and 70% respectively. The sensitivity with BTA, BFP and NMP22 for urothelial cancer was higher than that with urine cytology. However, all except for urine cytology showed high false positive rates (83-90%) for urinary tract infection. These markers may thus complement urine cytology, which has a low sensitivity for urothelial cancer. Quite possibly they could act as low-cost and useful tumor markers, which could in turn reduce the number of invasive cystoscopic examinations. However, considering their high false positive rates for benign disease such as urinary tract infection, we must acknowledge that an ideal urothelial tumor marker, which is simple, non-invasive, inexpensive and accurate with high sensitivity and specificity has yet to be developed. PMID- 11729493 TI - The risk manager's role in disclosure of medical error: seeing ourselves as others see us. PMID- 11729494 TI - Key considerations on drafting a policy on disclosure of unanticipated outcomes. PMID- 11729495 TI - Our experience in developing a non-punitive culture and an effective amnesty policy. PMID- 11729496 TI - Safety by design: ten lessons from human factors research. PMID- 11729497 TI - Meeting the challenges of patient safety through the design of a new risk management process. PMID- 11729498 TI - Effecting cultural change: Memorial Hospital's experience. PMID- 11729499 TI - Enhancing patient safety through teamwork training. AB - The effective reduction of medical errors depends on an environment of safety for patients in both clinically-based and systems-oriented arenas. Formal teamwork training is proposed as a systems approach that will achieve these ends. In a study conducted by Dynamics Research Corporation, weaknesses and error patterns in Emergency Department teamwork were assessed, and a prospective evaluation of a formal teamwork training intervention was conducted. Improvements were obtained in five key teamwork measures, and most importantly, clinical errors were significantly reduced. PMID- 11729500 TI - Disclosure of medical error: facts and fallacies. PMID- 11729501 TI - Community report cards--why disclose? PMID- 11729502 TI - Lovastatin increases nitric oxide synthesis in IL-1 beta-stimulated smooth muscle cells. AB - OBJECTIVE: Nitric oxide (NO) production by inducible NO synthase (iNOS) may play an important role in the pathogenesis of atherosclerosis. Although lovastatin has been shown to reduce the progression of atherosclerosis, it is not known whether it regulates NO production. We investigated the effects of lovastatin on NO synthesis and the mechanisms by which lovastatin exerts its effects in rat vascular smooth muscle cells. METHODS: Primary cultures of the vascular smooth muscle cells were obtained from the media of the thoracic aorta of Sprague Dawley rats (200-250 g). Nitrite levels in the culture medium of rat vascular smooth muscle cells were determined colorimetrically. RESULTS: Lovastatin (10(-5) mol/L) significantly increased interleukin-1 beta (IL-1 beta, 10 ng/mL)-induced nitrite accumulation in a time (0-24 hours)-dependent manner. Exogenous mevalonate and geranylgeranyl-pyrophosphate completely reversed the stimulatory effects of lovastatin on nitrite production. Furthermore, inhibition of Rho by C3 exoenzyme mimicked the increase in IL-1 beta-induced nitrite accumulation induced by lovastatin in the vascular smooth muscle cells. CONCLUSION: These results demonstrate that lovastatin up-regulates NO formation in rat vascular smooth muscle cells stimulated by IL-1 beta, and the effect may be associated with the inhibition of Rho activity. PMID- 11729503 TI - Nationality differences in distributions of serum lipids, lipoproteins and apolipoproteins levels in Xinjiang China. AB - OBJECTIVE: To reveal the distribution characteristics of serum lipids, lipoproteins, and apolipoproteins levels in different nationalities. METHODS: Quantitative levels of those traits mentioned above were determined and body height (H), weight (W) and body mass index (BMI = W/H2) were assessed in 773 Kazaks (men 360 and women 413) and 911 Han nationality (men 466 and women 445) from the Xinjiang Autonomous Region of China. RESULTS: Kazaks men and women, respectively, had significantly higher serum levels of HDL-c and ApoA1 (all P < 0.001) and significantly lower serum levels of TG (men P < 0.01, women P < 0.001), Lp(a) (all P < 0.005), ApoB (men P < 0.005, women P < 0.001) and ApoB/A1 (men P < 0.01, women P < 0.001) than their Han nationality controls. The threshold points of ApoA1 (< 1.2 g/L) and ApoB (> 1.2 g/L) are higher in Hans than in Kazaks for ApoA1 (20.7 vs 9.8%) and ApoB (18.6 vs 14.3%); the serum levels of TG, HDL-c, Lp(a), ApoA1 and ApoB/A1 are all closely correlated with nationality (P < 0.001, P = 0.001, and P < 0.05, respectively). CONCLUSION: The results suggest that the nationality differences exist in serum levels of lipids, lipoproteins, and apolipoproteins, and Kazaks have a superior serum lipid pedigree to Hans. This differences may come from genetic differences, which affect the serum levels of lipids, lipoproteins, and apolipoproteins by controlling lipid metabolism patterns. Future study will be needed to dissect to the role of genetic factors on serum lipids. PMID- 11729504 TI - Abnormal metabolism of nitric oxide, oxidative stress and lipoperoxidative stress in patients with acute viral myocarditis. AB - OBJECTIVE: To investigate metabolic status of nitric oxide (NO) as well as oxidative and lipoperoxidative stress, pathological chain reactions of a series of free radicals, and oxidative and lipoperoxidative damages in patients with acute viral myocarditis (AVM). METHODS: Using a random paired control design, plasma levels of NO, lipoperoxides (LPO), vitamin C (VC), vitamin E (VE) and beta carotene (beta-CAR), erythrocytic level of LPO as well as activities of erythrocytic superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were investigated by spectrophotometric assays in 60 AVM patients and 60 healthy adult volunteers (HAV). RESULTS: Compared with the HAV group, plasma NO, and LPO in plasma and in erythrocyte of the AVM group significantly increased (P = 0.0001), while VC, VE, beta-CAR, SOD, CAT and GSH-Px of the AVM group significantly decreased (P = 0.0001). Compared with the AVM group before treatment, plasma NO, and LPO in plasma and in erythrocyte of the AVM group post treatment significantly decreased (P < 0.05), whereas plasma VC, VE and beta-CAR as well as erythrocytic SOD, CAT and GSH-Px of the AVM group post treatment significantly increased (P < 0.05). CONCLUSION: The findings in this study suggested that in the AVM patients the metabolism of NO was disturbed, and the pathological chain reactions of a series of free radicals were severely aggravated, thus produced the oxidative damage and lipoperoxidative damages. Therefore, we recommend that antioxidants at suitable dosage, such as VC, VE, beta-CAR and others, should be given to AVM patients daily to alleviate potential oxidative and lipoperoxidative damages in their bodies. PMID- 11729505 TI - Percutaneous transluminal stenting in patients with carotid artery stenosis. AB - OBJECTIVE: To assess the efficacy and safety of percutaneous transluminal stenting for patients with carotid artery stenosis. METHODS: Selective percutaneous transluminal stenting was performed for patients with symptomatic carotid artery stenosis (luminal narrowing > or = 70%). Success rates and complications associated with the procedures were observed. During six months of follow-up, both recurrent symptom and restenosis rates were recorded. RESULTS: There were 17 bifurcating lesions among 27 stenoses in 26 patients, of whom 18 had concomitant coronary artery diseases. The acute procedural success rate was 96.3% (26/27), and the degree of stenosis was reduced from 88.6% +/- 8.9% (range 70-100) to 0.4% +/- 2.0% (range 0-10). Six patients developed severe bradycardia and hypotension, and 3 experienced transient loss of consciousness during balloon dilatation. During hospitalization, 2 patients experienced loss of consciousness and convulsion, respectively, due to hyperperfusion, and both recovered 12 hours later. There were 2 minor stroke cases (7.4%) but no cases of major stroke or death. At the 6-month follow-up, there were no cases of TIA or new onset of stroke. There was no restenosis detected in 16 cases using angiography and in 10 cases using MRI in 6 to 16 months of follow-up. CONCLUSIONS: Percutaneous transluminal stenting for patients with carotid artery stenosis has a high procedural success rate with few and acceptable complications. Few patients suffered from recurrent symptoms or showed restenosis in long-term follow-up. PMID- 11729506 TI - Interleukin-6 in bronchoalveolar lavage fluid from patients with COPD. AB - OBJECTIVE: To obtain new insights into the behavior of Interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) and released from alveolar macrophages (AM) in chronic obstructive pulmonary diseases (COPD), and reveal the relationship between IL-6 and the development of emphysema in COPD. METHODS: IL-6 in BALF and released by AM in BALF were examined in 7 non-smoking subjects and 21 patients with COPD. According to the 95% confidence limits of IL-6 in BALF from non smoking subjects, the patients were divided into two groups: those who were within the limits were assigned to the first group, and those who were above the limits were assigned to the second group. RESULTS: The concentration of IL-6 released by AM was much higher in the second group than in the first one. Between the two groups, significant differences were found in pulmonary function. CONCLUSION: Our results suggest that the concentration of IL-6 released by AM may be related with pulmonary function, and IL-6 may play a role in the development of emphysema in patients with COPD. PMID- 11729507 TI - Alteration of surfactant proteins A and D in bronchoalveolar lavage fluid of Pneumocystis carinii pneumonia. AB - OBJECTIVE: To understand the interaction between surfactant proteins and pneumocystis carinii pneumonia (PCP), and the impact of corticosteriods on surfactant proteins. METHODS: We established rat models of PCP and bacterial pneumonia induced by subcutaneous injection of 25 mg cortisone acetate. At 8-12 wk, the bronchoalveolar lavage fluid (BALF) of rats was collected. Total nucleated cells of BALF were counted and differentiated, and the concentrations of surfactant protein A (SP-A) and surfactant protein D (SP-D) were measured by immunoblotting assay. The rats were divided into three immunosuppressive groups and a normal control group. Group I, normal control (n = 6), consisted of healthy SD rats; group II, negative control (n = 6), consisted of rats with cortisone acetate injection for over 8 wk without lung infection; group III, bacterial pneumonia (n = 11), rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated; and group IV, PCP (n = 14), rats with injected cortisone acetate for 8-12 wk and developed PCP without other pathogens isolated. RESULTS: Our results indicated that the total cell count in BALF in the negative control group was lower than that in the normal control group (P < 0.001). During PCP infection, the total cell count and the percentage of polymorphonuclearcytes (PMNs) in BALF were significantly increased (P < 0.01), but were lower than those in the bacterial pneumonia group. The concentration of SP-A of BALF in PCP (45.1 +/- 22.1 micrograms/ml) was significantly increased in comparison with that in the negative control (16.2 +/- 9.9 micrograms/ml, P < 0.05) and bacterial pneumonia groups (6.2 +/- 5.6 micrograms/ml, P < 0.001). We also found that the relative content of SP-D was significantly higher in PCP (24,249 +/- 4780 grey values) than that in the negative control (13,384 +/- 2887 grey values, P < 0.001) and that in bacterial pneumonia (11,989 +/- 2750 grey values, P < 0.001). SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group (P < 0.01, P < 0.001). SP-A and SP-D were higher in the negative control group than those in the normal control group, but there was no significant difference between the 2 groups. CONCLUSION: These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation, but the alteration is not related to the corticosteriod usage. PMID- 11729508 TI - An automated fluorescent single strand conformation polymorphism technique for high throughput mutation screening. AB - OBJECTIVE: To develop a high throughput mutational detection method by multiple fluorescence-labeled polymerase chain reaction (PCR) products. METHODS: A total of 27 known mutations including 22 substitutions, 3 insertions (1, 2 and 7 bp) and 2 deletions (1 and 2 bp) in the hepatocyte nuclear factor (HNF)-4 alpha, glucokinase and HNF-1 alpha genes were tested. During nested PCR, amplified fragments were labeled with three fluorescent dyes. PCR products were visualized with an ABI-377 fluorescence sequencer using 5% glycerol or 10% sucrose in non denaturing gel conditions. RESULTS: Twenty-five of 27 variants (93%) could be detected by combining 5% glycerol and 10% sucrose gel matrix conditions. Twenty two of 27 (82%) and 18 of 27 (67%) variants were identified using 5% glycerol and 10% sucrose conditions, respectively. CONCLUSION: This fluorescence-based PCR single strand conformation polymorphism technique represents a simple, non hazardous, time-saving and sensitive method for high throughput mutation detection. PMID- 11729509 TI - Effects of prolactin on HLA-DR and CD40 expressions by human thyrocytes. AB - OBJECTIVE: To study the effects of prolactin (PRL) on HLA-DR and CD40 expressions by human thyrocytes, and to investigate the possible mechanisms for PRL to affect the development of Graves' disease (GD). METHODS: Thyrocytes in secondary culture, which were from GD thyroid glands and the tissues adjacent to the lesions of multinodular goiter and adenoma (control group), were treated respectively with ovine PRL (oPRL), interferon-gamma (IFN-gamma), interlukin-4 (IL-4) and oPRL plus IFN-gamma (10 U/ml) or IL-4 (5 ng/ml) for 7 days. HLA-DR and CD40 expressions on the thyrocytes were determined by immunofluorescent staining and flow cytometry. RESULTS: oPRL (12.5 ng/ml-1000 ng/ml) had no significant direct effect on HLA-DR or CD40 expression. It did not significantly affect the stimulation of HLA-DR expressions on the two groups of thyrocyte treated with IFN gamma or on GD thyrocytes treated with IL-4. oPRL could antagonize the stimulation of CD40 expressions by IFN-gamma and the inhibition by IL-4 on both groups of thyrocytes. The antagonizing effects were related to the concentrations of PRL. IFN-gamma-stimulated percentages of CD40+ thyrocytes and delta mean fluorescence intensity (dMF) in both thyrocyte sources were significantly reduced in the presence of 200 ng/ml oPRL (both GD and CONTROL: P < 0.01 and P < 0.05, respectively; CONTROL: P < 0.05) and 1000 ng/ml oPRL (GD: P < 0.01; CONTROL: P < 0.05). oPRL caused significant increasing in IL-4-inhibited percentages of CD40+ cells from the two groups of thyrocytes at 12.5 ng/ml and 1000 ng/ml and dMF from GD thyrocytes at 1000 ng/ml (P < 0.05). CONCLUSIONS: PRL can exert indirect effects on CD40 expressions on thyrocytes by antagonizing the modulatory actions of IFN-gamma and IL-4 with dose-related effects. This may be one important mechanism for PRL to affect the development of GD. PMID- 11729510 TI - A retrospective study of continuous renal replacement therapy versus intermittent hemodialysis in severe acute renal failure. AB - OBJECTIVE: To investigate the efficacy of continuous renal replacement therapy (CRRT) versus intermittent hemodialysis (IHD) in patients with severe acute renal failure (ARF). METHODS: One hundred and ninety-three severe ARF patients who received renal support between December 1978 and December 1998 were involved in this study. Of them, 101 (52.3%) were treated with CRRT (CRRT group), and 92 (47.7%) with IHD (IHD group). RESULTS: Sixty (59.4%) patients in the CRRT group got through the acute phase of disease and 41 (40.6%) patients did not survive while in the IHD group 59 (64.1%) patients survived and 33 (35.9%) patients did not. No significant difference in survival rate was found between the two groups. 24 of 64 patients (37.5%) in the CRRT group with multiple organ dysfunction syndrome (MODS) survived, while in the IHD group, 8 out of 44 (27.3%) survived, their survival rate was much lower than that in the CRRT group. Patients in CRRT group were more severely ill, as manifested by lower mean arterial pressure, higher APACHE II score, more dysfunctioned organs and requiring mechanical ventilation and vasopressor support as compared with patients in the IHD group, CRRT was found to improve hemodynamic stability with a better fluid balance and control of biochemical status, increased nutritional intake and a shorter duration of acute renal failure (P < 0.05). CONCLUSION: CRRT perhaps may be the best choice in the treatment of severe ARF patients, for it can offer several distinct advantages compared to IHD. These may contribute to improving the survival rate of ARF patients, particularly those that are critically ill patients. PMID- 11729511 TI - Genotypes and polymorphisms of mutant CCR5-delta 32, CCR2-64I and SDF1-3' a HIV-1 resistance alleles in indigenous Han Chinese. AB - OBJECTIVE: To evaluate the frequencies and polymorphisms of CCR5-delta 32, CCR2 64I and SDF1-3' A alleles conferring resistance to HIV-1 infection in Chinese population from Han ethnic origin. METHODS: This cohort was comprised of 1251 subjects (915 men and 336 women) aged 15-80 years and none was HIV-1 positive. Genotyping of allelic CCR5-delta 32, CCR2-64I and SDF1-3' A variants was performed using PCR or PCR/RFLP assay, and further confirmed by direct DNA sequencing. RESULTS: Our finding shows that the delta 32 deletion mutation in the CCR5 gene does occur in this population and can be inherited in a Mendelian fashion in indigenous Han Chinese at a very low frequency of 0.00119 (n = 1254). The frequencies of mutant CCR2-64I and SDF1-3' A alleles were 0.20023 (n = 1251) and 0.2873 (n = 893), in this population, which are higher than those found in American Caucasians. Furthermore the polymorphisms of CCR2-64I and SDF1-3' A alleles in the Han Chinese population were different from those in American Caucasians. Statistical analysis showed that the genotype distribution of CCR5 delta 32, CCR2-64I and SDF1-3' A alleles was in equilibrium according to the Hardy-Weinberg equation. CONCLUSION: The CCR5-delta 32 mutation may not be a major resistant factor against HIV-1 infection in indigenous Han Chinese. The significance of higher frequencies of CCR2-641 and SDF1-3' A alleles (0.20023 and 0.2791) in the Han population remains to be clarified in HIV-1-positive carriers and AIDS patients. PMID- 11729512 TI - Antineoplastic mechanism of Octreotide action in human hepatoma. AB - OBJECTIVES: To investigate whether apoptosis can be induced by Octreotide in human hepatoma cells in vitro and elucidate the antineoplastic mechanism of Octreotide in hepatoma. METHODS: A cultured human hepatoma cell line, BEL-7402, was exposed to Octreotide and apoptosis was evaluated by cytochemical staining (Hochesst 33,258), transmission electron microscopy, agarose gel electrophoresis and flow cytometry (FCM). RESULTS: After exposure to 0.2 microgram/ml Octreotide, apoptosis with nuclear chromatin condensation as well as fragmentation, cell shrinkage and the formation of apoptotic bodies was observed using cytochemical staining and transmission electron microscopy. A DNA ladder in agarose gel electrophoresis was also displayed. FCM showed that the apoptotic cell number rose with an increase in the concentration of Octreotide (0-2 micrograms/ml). There was a positive correlation between Octreotide concentration and apoptotic rate in BEL-7402 cells (r = 0.809, P < 0.05). CONCLUSION: Apoptosis in human hepatoma cells can be induced by Octreotide, which may be related to the mechanism of antineoplastic action of Octreotide in hepatoma. PMID- 11729513 TI - Fructose-1,6-diphosphate-added total parenteral nutrition in septic animals and stressed patients. AB - OBJECTIVE: To investigate the roles of fructose-1,6-diphosphate (FDP)-added total parenteral nutrition (TPN) in septic animals and stressed patients. METHODS: Thirteen adult dogs were randomly assigned to one of two study groups 6 hours after the induction of severe intra-abdominal infection. Group TPN (n = 6) received 70 kcal.kg-1.d-1 of nonprotein calorie (NPC) and 0.56 g.kg-1.d-1 of nitrogen. 1 g/kg of FDP was also infused to the animals in group TPN + FDP (n = 7) everyday. In the clinical study, the control group received routine TPN, while the study group (n = 16) was treated with TPN plus FDP (5 g, two times a day) for 7 days. RESULTS: In dogs with TPN support, plasma ATP levels were not changed significantly, while the value in the TPN + FDP group increased significantly from 0.18 mumol/L to 0.46 mumol/L at 24 h and 0.51 mumol/L at 48 h (P < 0.01). Muscular ATP increased markedly in the TPN + FDP group. Muscular creatine phosphate values were not significantly changed in the TPN group, but the values increased in the TPN + FDP group from 4.06 mumol/g.wt at the beginning to 4.93 mumol/g.wt at 24 h and 5.60 mumol/g.wt at 48 h (P < 0.05), with a cytochrome oxidase increase in immunohistochemistry stain. In the clinical study, plasma ATP levels increased and urinary 3-methylhistidine production significantly decreased with an improved value for positive accumulative nitrogen balance in the FDP infused group. CONCLUSION: Our results suggest that total parenteral nutrition support with the supplement of fructose-1,6-diphosphate has a positive role in body energy production and protein metabolism in septic animals and stressed patients. PMID- 11729514 TI - Aortic root replacement. AB - OBJECTIVE: To report our operative experience with aortic root replacement in 231 patients with aortic root aneurysm and discuss the current indications, methods, and surgical techniques. METHODS: Between January 1994 and August 1999, a group of 231 consecutive patients underwent aortic root replacement at our hospital, with 13 being treated on an emergency basis. There were 189 men and 42 women, ranging in age from 14 to 69 years. The diameter of the aneurysm varied from 4.5 to 11 cm. Among this group, 145 had isolated aortic root aneurysms, 65 suffered from DeBakey type I aortic dissection, and the remaining 21 were diagnosed as having DeBakey type II aortic dissection. Aortic valve regurgitation occurred in all cases. Aortic root replacement was performed with composite valved graft in 229 patients, and in 2 patients the aortic valve was preserved. RESULTS: The hospital mortality rate was 3.03% (7 patients). Early complications included re exploration for bleeding in 6 patients, pericardial effusion in 9, as well as cerebral infarction, pleural effusion, and pneumothorax in 2 patients each. One hundred and seventy-five patients (78.12%) were followed up, with a mean follow up time of 15.7 +/- 13.1 months (range, 2 weeks to 65 months). One patient died from lower-limb embolism and renal dysfunction 3 months postoperatively. Three patients died from postoperative anticoagulation accidents. The preoperative and postoperative mean left ventricular end-diastolic diameters were significantly different (68.1 +/- 9.4 mm, range 54 to 112 mm; vs 54.8 +/- 8.2 mm, range 38 to 88 mm; P < 0.001). CONCLUSIONS: Once a diagnosis of acute aortic root dissecting aneurysm is made, the patient should undergo surgery as soon as possible if the general conditions permit. Aortic aneurysm without dissection or with chronic dissection should be operated if the diameter of the aneurysm is greater than 5 cm. PMID- 11729515 TI - Aortic and mitral valve replacement with retrograde perfusion in the beating heart. AB - OBJECTIVE: To estimate the value of aortic valves and combined mitral valve replacement with retrograde perfusion in beating hearts. METHODS: Continuous retrograde coronary sinus perfusion with beating hearts was used in 83 patients undergoing aortic valve or aortic valve combined with mitral valve replacement, without application of cardioplegia. After aortic valve replacement, the retrograde perfusion was changed to antegrade perfusion for mitral valve replacement or correction of the other deformities (group A). Cold blood cardioplegia solution (15 degrees C) was infused at intervals in 20 cases (group B). The following parameters were tested: lactate, ET, CTn-T and MDA in blood; myocardial ultra-structure; and cardiac rhythm and cardiac output (CO). RESULTS: All biochemical values increased after cardiopulmonary bypass (P < 0.05-0.01). Empty and beating heart sinus rhythm was maintained in group A. Myocardial ultrastructure did not change significantly. The pump was stopped smoothly as the surgical procedure finished. No postoperative low cardiac output syndrome or arrhythmia was observed. Eight-one patients recovered smoothly, two died from renal failure or infective shock. When the pump stopped, all patients in group B were supported by 5-10 micrograms.kg-1.min-1 dopamine. Transient pacing was used in 9 patients. One patient died from low cardiac output syndrome. CONCLUSION: This method is a good myocardial protection which simulates physiologic status. It is applicable to aortic valve and combined mitral valve replacement of patients with large heart or heart failure and long time aortic cross-clamping. Ideal clinical effect can be achieved. PMID- 11729516 TI - Myocardial protection of immature rabbits with an ATP-sensitive K+ channel opener pinacidil. AB - OBJECTIVE: To investigate the effectiveness of pinacidil, an opener of ATP sensitive K+ channels, in protecting the myocardium of immature rabbit hearts from ischemic reperfusion injury. METHODS: Rabbit hearts underwent 30 min of global normothermic ischemia followed by 30 min of reperfusion on the modified Langendorff apparatus. Fifty-two isolated hearts of 3-4 week-old immature rabbits were divided into 4 groups randomly. During ischemia, 3 different cardioplegic solutions were administered intermittently by infusion every 15 min (20-25 ml each time in all groups). Group 1: control group (n = 13); group 2: Krebs Henseleit (K-H) solution with potassium (16 mmol/L) (n = 13); group 3: K-H solution with potassium (16 mmol/L) and pinacidil (50 mumol/L) (n = 13); group 4: K-H solution with potassium (16 mmol/L), pinacidil (50 mumol/L) and glibenclamide (10 mumol/L) (n = 13). The pre-ischemic and post-ischemic myocardial functions were assessed by the percentage recovery of the left ventricular developed pressure (LVDP); the left ventricular end-diastolic pressure (LVEDP); both the positive peak and negative peaks of the first derivative of the left ventricular pressures (+/- dp/dtmax); coronary flow; the level of creatine kinase (CK), lactic dehydrogenase (LDH) and aspartate transcarbamoylase (AST) in coronary sinus venous effluent; and by myocardial ultrastructural changes. RESULTS: Before myocardial ischemia, there were no significant differences among the four groups in any of the parameters mentioned above. Post-ischemic recovery of LVDP, LVEDP, +/- dp/dtmax, coronary flow, the level of CK, LDH and AST, and myocardial ultrastructural changes were better in group 3 than those in the three other groups. CONCLUSIONS: As a new and effective composition, pinacidil can significantly improve myocardial protection from cardioplegia for immature rabbit hearts. PMID- 11729517 TI - A histomorphometric and molecular study on stress adaptability of freeze-dried bone allograft. AB - OBJECTIVE: To Investigate stress adaptability of freeze-dried bone allograft. METHODS: Cortical and cancellous allograft were transplanted to each side of the midshaft diaphyseal ulna in two groups of 28 animals. The left transplanted allograft was free from fixation and bore a normal physiological load, while the right transplanted allograft was protected from loading by a simple external fixator and bore less load. Animals were sacrificed at the 2nd, 4th, 8th, 16th week after transplantation and specimens were taken out for bone histomorphometry studies and analysis of collagen gene expression by in situ cDNA-mRNA hybridization. RESULTS: Labeled surface(LS) and bone mineral apposition rate(MAR) of the normally loaded graft-host bone interface were significantly higher than that of the less loaded side at the 4th, 8th, 16th week after transplantation. Parameters reflecting the internal repair process of the allograft, such as LS in cortical and cancellous bone or MAR in cortical bone of the normally loaded side were significantly higher than those of the less loaded side at the 16th week after transplantation. The result of in situ hybridization indicated that more osteoblast-like cells expressing the type I collagen gene were found in the interface or interior of normally loaded grafts. CONCLUSION: The stimulus of physiologic load can accelerate the early union of allograft-host bone interface and later new bone creep substitution to the necrotic allograft. PMID- 11729518 TI - Characteristics of spongiform leukoencephalopathy induced by heroin: MRI detection. AB - OBJECTIVE: To elucidate the imaging characteristics of spongiform leukoencephalopathy induced by heroin and the diagnostic value of MRI examination. METHODS: Eleven cases of heroin addiction were examined by MRI. The sequences included T1-weighted imaging (T1WI), T2-weighted imaging (T2WI) and fast fluid-attenuated inversion recovery pulse sequence (Flair). RESULTS: Bilateral symmetrical lesions were located in the white matter of the cerebrum and cerebellum, the internal capsule in all cases, and the callosum in 6 cases. Elliptical lesions in the mesencephalon were found in two cases. These lesions were shown as hypointense signals on T1WI and hyperintense signals on T2WI and Flair's imaging. Cerebellar white matter suffered more severely, but the dentate nuclei were not invaded. CONCLUSION: Heroin-induced spongiform leukoencephalopathy has characteristic patterns in MRI, which are very specific to this form of the disease. PMID- 11729519 TI - Nasal carriage of Streptococcus pneumoniae among children in Beijing. AB - OBJECTIVE: To investigate the antimicrobial susceptibility of Streptococcus pneumoniae carried in the nose among children in Beijing and the distribution of serotypes, and to analyze the risk factors for nasal carriage of penicillin non susceptible S. pneumoniae. METHODS: A disk diffusion test was applied to detect the antimicrobial susceptibilities of S. pneumoniae to erythromycin, trimethoprim sulfamethoxazole, chloramphenicol and tetracycline. The E test was applied to determine the minimal inhibitory concentrations of penicillin, cefuroxime, cefotaxime, augmentin and imipenem. S. pneumoniae isolates were serotyped by the Quellung reaction. RESULTS: S. pneumoniae that was resistant to penicillin or cefuroxime was not found, but S. pneumoniae intermediate resistant to penicillin and cefuroxime accounted for 8.2% and 2.1%, respectively. All of the isolates were susceptible to cefotaxime, augmentin and imipenem. S. pneumonia that was resistant to erythromycin, trimethoprim-sulfamethoxazole and tetracycline were extremely numerous, accounting for 72%, 70% and 79%, respectively. Five serotypes (19, 6, 14, 23, 17) accounted for 54.7%, and nontypables accounted for 20.6% of all the S. pneumoniae. Previous history of otitis media was a risk factor we found for nasal carriage of penicillin non-susceptible S. pneumoniae. CONCLUSIONS: Continued surveillance of the antimicrobial susceptibilities of S. pneumoniae is necessary. A larger scale investigation is needed to identify if the 7 or 9-valent conjugate pneumococcal vaccine is appropriate for Chinese children. PMID- 11729520 TI - Enteric transmission of transfusion-transmitted virus. AB - OBJECTIVE: To detect the virus in the feces and sera of patients in an outbreak of enterically transmitted non-A, non-E hepatitis, and this review covers the epidemiologic features and experimental infection of this novel virus. DATA SOURCES: Data sources come from our own work on this subject, published and unpublished. STUDY SELECTION: Mainly our own work is included, and related literature is collected. RESULTS: In an outbreak of enterically transmitted non-A E hepatitis among students, a total of 381 cases (60.7%) were documented. Viral fragments identical to transfusion transmitted virus (TTV) were detected in both serum and stool samples. Asymptomatic virus carriers among the staff had positive serum (32.1%) and feces (24.6%), clearly a potential source of infection. This viral infection prevalence in 2 remote villages in northern and southern China was 9.2% and 10.6%, respectively, suggesting that China is an endemic area. In this study, groups of 3 Rhesus monkeys were infected via oral or intravenous inoculation with patient feces. Two additional monkeys were infected by passage. The virus was detected in serum, peripheral blood mononuclear cell (PBMC), liver, spleen and small intestine, while the virus positive single strand, which might be a replicative intermediate, was only in liver, intestine and PBMC of all animals. CONCLUSIONS: This nonenveloped DNA virus might be transmitted both by blood and enteric routes. Considering its wide distribution and high prevalence, we suppose that nonparenteral transmission is more important. PMID- 11729521 TI - Prevalence and clinical characteristics of mitochondrial tRNA leu(UUR) mt 3243 A- >G and ND-1 gene mt 3316 G-->A mutations in Chinese patients with type 2 diabetes. AB - OBJECTIVE: To assess the prevalence of mitochondrial tRNA leu(UUR) gene mt 3243 A ->G mutation and ND-1 gene mt 3316 G-->A mutation in Chinese type 2 diabetes. METHODS: 716 randomly selected, unrelated patients with type 2 diabetes were screened for the mutation with a PCR-RFLP technique. RESULTS: Three individuals with mitochondrial tRNA leu(UUR) gene mt 3243 A-->G mutation were identified, representing approximately 0.4% of the type 2 patients screened. Further screening of first-degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had: 1) an earlier diagnosis age of diabetes (38.0 +/- 10.1 year vs 53.4 +/- 10.0 year, P < 0.001); 2) lower Body Mass Index (BMI) (19.5 +/- 2.0 kg/m2 vs 24.9 +/- 10.9 kg/m2, P < 0.0001); and 3) lower post-challenge insulin levels (Area under the curve of insulin levels during the Oral Glucose Tolerance Test (OGTT), 2946 +/- 1647.2 microIU.ml 1 vs 7469 +/- 6647.7 microIU.ml-1, P < 0.01). ND-1 gene mt 3316 G-->A mutation was found in 16 patients with type 2 diabetes (2.2%) and 5 out 181 controls (2.7%) with normal glucose tolerance. Therefore, the frequency of the mutation was not different in patients and controls. Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt 3316 mutation and those without it. CONCLUSIONS: In this large cohort of Chinese Type 2 diabetes, the prevalence of mitochondrial tRNA leu(UUR) gene mt 3243 A-->G mutation was 0.4%, and the ND-1 gene mt 3316 G-->A mutation is a polymorphism unrelated to diabetes. PMID- 11729522 TI - Prevalence of CMV in arterial walls and leukocytes in patients with atherosclerosis. AB - OBJECTIVE: To investigate the association of cylomegalovirus with atherosclerosis. METHODS: The presence of cylomegalovirus (CMV) nucleic acids was demonstrated in carotid and coronary arteries of patients with and without atherosclerosis by polymerase chain reaction (PCR). CMV was detected by PCR in the blood of patients with and without atherosclerosis. RESULTS: 83.3%-86.7% of the samples obtained from atherosclerotic patient arterial walls were shown to contain viral nucleic acids (CMV), CMV could be found among 6.7% of patients' arterial wall without atherosclerosis, significant difference can be found between them (P < 0.01). In blood CMV could be found in 42.4% of patients with atherosclerosis, in the control group, only 3% of samples had CMV, P < 0.01. CONCLUSION: CMV plays an important role in the pathologic process of the atherosclerosis and the atherosclerotic cerebral infarction. PMID- 11729523 TI - Effect of thinking on intracranial blood flow velocities. AB - OBJECTIVE: To determine the effect of thinking on intracranial blood flow velocities. METHODS: Transcranial Doppler (TCD) was used to measure the blood flow velocities of the cerebral arteries, when 10 normal persons were at rest and during thinking. RESULTS: The mean flow velocities (MFVs) of the cerebral arteries during thinking were significantly higher than those at rest, especially in the MCA and ACA. The MFVs were raised from between 10.3% to 14.3%. CONCLUSIONS: Thinking can increase the blood flow velocities of cerebral arteries significantly. PMID- 11729524 TI - Vitamin D receptor gene polymorphism and bone mineral density in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To explore the relationship between vitamin D receptor (VDR) gene polymorphisms and bone mineral density (BMD) in patients with type 2 diabetes mellitus (DM) and to better understand the pathogenesis of osteoporosis. METHODS: Ninety seven patients with type 2 DM were recruited for this study. BMD was measured by single photon absorptiometry at the lower one third of the nondominant radius and ulna. Polymorphisms of the VDR gene were analyzed by DNA amplification with polymerase chain reaction (PCR) and endonuclease digestion with Bsm I. RESULTS: The respective frequencies of VDR genotypes were BB 18.6%, Bb 27.8% and bb 53.6%. The Z scores of the three groups were -1.57 +/- -0.60, 1.45 +/- -0.67 and -1.41 +/- -0.81, respectively. Although the BMD of the Bb genotype DM patients was higher than that of BB genotype DM patients and lower than that of bb genotype DM patients, there were no significant differences. CONCLUSION: These findings suggest a small influence of VDR gene polymorphism on the BMD of patients with type 2 DM. Further study on the value of VDR genotypes in the pathogenesis of osteoporosis in diabetes mellitus is still needed. PMID- 11729525 TI - Pseudohyperkalemia in thrombocythemia. PMID- 11729526 TI - Lymphangiomyomatosis. PMID- 11729527 TI - Relationships between hallucinations and disorganization in schizophrenic patients. AB - In two samples of 150 and 310 schizophrenic patients, scores for hallucinations were significantly associated with scores on disorganization. PMID- 11729528 TI - Career-related correlates of self-discipline. AB - An investigation was made of the relationship between scores on the Self discipline Questionnaire and self-ratings on four career-related variables among a sample of 325 working adults. The Self-discipline Questionnaire was previously published in a trade book and is based on characteristics of self-disciplined people culled from the literature. Scores for self-discipline were significantly correlated with years of formal education, salary, and self-perceptions of career success and frequency of goal accomplishment. PMID- 11729529 TI - Disgust sensitivity and attitudes toward organ donation among African-American college students. AB - 241 African-American college students (94 men and 147 women, mean age = 20.3 +/- 3.4 yr.) completed the 1994 Disgust Scale of Haidt, McCauley, and Rozin and a modified form of Parisi-Rizzo's 1987 Attitudes Toward Organ Donation Scale (negative subscale only) as well as a behavioral measure of intention to donate organs after death. Analyses indicated that the higher the disgust sensitivity, the more negative the attitude toward organ donation and the less likely the student was to indicate intent to donate organs. It was further shown that negative attitudes toward organ donation mediated the relationship between disgust sensitivity and the behavioral intention measure. Results highlight the complexity of the issues surrounding organ donation within an African-American population and provide additional empirical evidence for the development of a theoretical model to explain the organ donation phenomenon. PMID- 11729530 TI - Children's social competence, academic competence, and aggressiveness as related to ability to make judgments of fairness. AB - The relationship of 102 fifth and sixth graders' judgments of fairness with their academic and social competence and aggressiveness was studied. Children were given self-report and peer-nomination inventories measuring academic competence, social competence, and aggressiveness at a single point in time at the public school they attended. These measures were related to children's prediction of fairness on a distributive justice measure. Children who saw themselves as academically and socially competent scored higher on the distributive justice measure. Children whose peers saw them as less academically competent and more aggressive scored lower on the distributive justice measure. Analyses showed an association between children's academic competence, social competence, and aggressiveness scores and their scores on judgments of fairness. PMID- 11729531 TI - The psychological health of "joiners". AB - The current study examined the relationship between scores on social complexity and psychological health. A sample of 202 college students yielded positive correlations for propensity to join and maintain memberships in diverse groups with psychological health, thereby replicating past research. PMID- 11729532 TI - Predicting the extent and stressfulness of problem rumors at home among Army wives of soldiers deployed overseas on a humanitarian mission. AB - Nearly 500 civilian wives of enlisted soldiers from Fort Drum, New York were surveyed about the extent to which they experienced, as problems, rumors about what was happening in Somalia and when their soldier would return from the Operation Restore Hope deployment to Somalia in 1993. Rumors, as problems, were cited less frequently than loneliness, fears about the soldier's safety, or ignorance of the situation in Somalia but more frequently than problems with communication, finances, or legal issues. The strongest initial predictor of such problems was having communication problems with the soldier. Other significant predictors included length of deployment, soldier's rank, and unit support systems. More frequent use of telephone or regular mail, however, did not appear to reduce such rumors. Stressfulness of rumors appeared to be reduced by good unit leadership, good family support groups, and better emotional adaptability to deployment by spouses, while increased by reliance on surface mail for communication with one's deployed spouse. Subsequent analyses indicated that having fears about the safety of one's soldier or concerns about not knowing what was going on in Somalia were also significantly correlated with rumors as problems. Our results may support hypotheses that rumors serve to fill gaps in official information and justify spouses' anxieties regarding the safety of their deployed soldier. PMID- 11729533 TI - Understanding successful community living by individuals with serious and persistent mental illness. AB - 34 volunteers diagnosed as chronically mentally ill were administered a questionnaire to evaluate what factors were important in preventing rehospitalization. The volunteers were divided into two groups based on their relapse history. Analysis showed the nonrelapse group were more likely to attribute factors such as in-home therapy, taking medication, and having fun with friends than did the group who had experienced at least one relapse in the past 24 months. PMID- 11729534 TI - Ropelike birthmarks on children who claim to remember past lives. AB - Description of birthmarks having the pattern of strands of a rope in a second known case includes some verification of the correspondence between the birthmarks and injuries from ropes on an identified decreased person. PMID- 11729535 TI - A tract on psychology: agency and communion in human mentation. AB - Listed are 117 propositions, generated as beginning notes for development of theoretical understanding of agency and communion in human mentation. PMID- 11729536 TI - Recent research on family factors and readiness: implications for military leaders. AB - Past research on the relationship between family factors and military readiness in the Army has suggested that family issues affect retention far more than readiness. New data on individual soldier readiness were analyzed to assess the relative importance of family factors (internal and external family adaptation) compared to unit readiness, longevity, rank, and gender. Family factors were significant predictors, although external family adaptation appeared to be more important than internal family adaptation. Although variables related to retention were more strongly related to family factors than our readiness variables, the differences were less substantial than those reported previously. The data seemed to suggest that reserve-component readiness might be more affected by family stress than the active component and that officer readiness might be more strongly affected by family worries than enlisted personnel readiness, but more research is needed to confirm those results. PMID- 11729537 TI - Eliminating invalid self-report survey data. AB - A sample of 6,370 students in Grades 6 to 8 completed a questionnaire on their attitudes and use of alcohol, tobacco, and other drugs. A subsample showed questionable data based on three criteria: missing responses, invalid responses, and inconsistent responses. Analysis indicated that this subsample was significantly different from the main group on demographic variables and self reported life-time tobacco use. Results support efforts to identify and eliminate invalid data. PMID- 11729538 TI - Outcome comparison of patients receiving oral or depot neuroleptic medication. AB - Three outcome measures of psychiatric treatment were retrospectively compared between two groups of acutely hospitalized psychotic patients who had been prescribed either oral haloperidol (n = 58) or depot haloperidol (n = 95) as the only medication at the time of discharge. There were no significant differences between the groups on demographic variables, including sex, age, and ethnicity. There were no significant differences between the groups on keeping the initial outpatient appointment, in readmission history over a 4-yr. follow-up, or in length of time to subsequent readmission, if any. It appears that there is no benefit in choosing one form of medication over the other from a clinical perspective, as neither appears more clinically efficacious when examined on these outcome variables. However, there is a significant difference in cost of the two forms of the medication in favor of the oral form, making the oral form more favorable if cost is to be minimized. PMID- 11729539 TI - Undertakers' sense of humor. AB - A group of 60 middle-aged morticians at a professional seminar in the midwestern USA who completed a multidimensional sense of humor scale scored significantly lower than another group of 136 men from other occupations. The difference between the two groups appeared almost entirely on scale items having to do with humor generation or creativity. PMID- 11729540 TI - Companion cats and the social support systems of men with AIDS. AB - 60 male pet owners with AIDS were given a formal questionnaire including items related to demographic, lifestyle, and pet ownership, as well as the Comfort from Companion Animals Scale, the Revised UCLA Loneliness Scale, and the General Health Questionnaire (GHQ-28). The men lived in the San Francisco area and received assistance from Pets Are Wonderful Support (PAWS), a community organization, in caring for their companion dogs or cats. Participants were a convenience sample of pet owners who volunteered from among 500 PAWS clients and who do not necessarily represent a random sample of men with AIDS who keep companion animals. For these men, their comfort from companion animals was significantly associated with having cats but not dogs, closeness with friends, dissatisfaction with their practical support system, and listing pets as a source of support. Loneliness was negatively associated with having a large practical support network, scoring as healthy on the General Health Questionnaire, and living alone. For these male pet owners with AIDS, pet cats appeared to complement their supportive relationships with friends and family that were protective against loneliness. PMID- 11729541 TI - Weight gain in post-seized rats is facilitated by adding aspirin, glucose, or glucose-taurine-acetaminophen to food mush. AB - Adult male rats were seized with lithium and pilocarpine and administered acepromazine to facilitate survival. After four days (1) 8 mg of acetylsalicylic acid (aspirin), (2) 100 mg taurine-15 mg acetaminophen (Tylenol)-40 mg glucose, (3) 40 mg glucose, or (4) water was added to the food mush daily for 30 days. A fifth group served as non-seized controls. Within one week all pharmacological treatments promoted more weight recovery than food mush only. The rats receiving aspirin (equivalent to 3 tablets/day for humans) showed the greatest early recovery. After 15 days of treatment the pharmacologically treated seized rats had returned to baseline weight and did not differ from normals whereas seized rats given only food mush had not. We suggest inhibiting prostaglandins by anti inflammatory compounds or stimulating the GABA shunt pathway through enhanced dietary glucose to accelerate weight gain following the significant loss that accompanies brain injury. PMID- 11729542 TI - Contribution of job satisfaction to happiness of Asian Americans. AB - Many demographic and labor force characteristics, such as family income, educational attainment, and occupation, correlated with job satisfaction. Since Asian Americans are more like Euro-Americans than African Americans in most of these characteristics, it seems reasonable to predict that their job satisfaction would be high as for Euro-Americans rather than low as for African Americans. Yet research of Weaver and Hinson showed that the opposite is true. One explanation for this unexpected result is that Asians do not think of jobs as a source of happiness but simply as a means of earning money to underwrite other aspects of their lives, such as the well-being of their families, which are the main sources of their happiness. The hypothesis was tested that job satisfaction does not contribute to the happiness of Asian Americans in comparison to satisfaction from other domains of their lives. Analysis was conducted of the attitudes of Asian American (n = 160), African-American (n = 602), and Euro-American (n = 6,477) workers who responded to 22 surveys drawn from 1972 to 1998, each of which was representative of the labor force of the USA. The hypothesis was supported by the finding that the partial correlation of job satisfaction and global happiness with satisfaction in seven other domains of life (marriage, financial condition, community, nonwork activities, family, health and physical condition, and friendships) held constant was significant for Euro-American women and men but not for Asian Americans or African Americans of either sex. And, the same result occurred when global happiness was regressed on job satisfaction net the effects of satisfaction in other seven domains. PMID- 11729543 TI - Complex partial epileptic-like experiences in university students and practitioners of Dharmakaya in Thailand: comparison with Canadian university students. AB - We tested the hypothesis that individuals who frequently practice meditation within another culture whose assumptions explicitly endorse this practice should exhibit more frequent and varied experience associated with complex partial epilepsy (without the seizures) as inferred by the Personal Philosophy Inventory and Roberts' Questionnaire for the Epileptic Spectrum Disorder. 80 practitioners of Dharma Meditation and 24 university students in Thailand were compared with 76 students from first-year courses in psychology in a Canadian university. Although there were large significant differences for some items and clusters of items expected as a result of cultural differences, there were no statistically significant differences between the two populations for the proportions of complex partial epileptic-like experiences or their frequency of occurrence. There were no strong or consistent correlations between the history of meditation within the sample who practiced Dharma meditation and these experiences. These results suggest complex partial epileptic-like experiences may be a normal feature of the human species. PMID- 11729544 TI - Verbal aggression in computer-mediated decision making. AB - 37 participants receiving verbally aggressive messages during a mediated interaction viewed their interactants as less socially attractive. PMID- 11729545 TI - Sex differences in associations of temperament with love-styles. AB - University students (439 women, 111 men) completed a questionnaire that included items assessing temperament and love-styles. While a single temperament and love style association characterized the men, all five temperament dimensions were predictive of certain of the love-styles for the women. PMID- 11729546 TI - Breast self-examination and cervical (Pap) smear test: attitudes and self reported practices among South African women. AB - This study investigated the attitudes and practices of breast self-examination and cervical (Pap) smear test among South African women. The sample included 150 Black and 150 White women chosen by systematic random sampling from telephone directories. The Black women were within the age range of 18 to 80 years (M age 38.9 yr., SD = 4.7), and the White women were within the age range of 19 to 91 years (M age 41.8 yr., SD = 3.5). Analysis indicated that 97.1% of the White women and 44.9% of the Black women said that they know how to examine their breasts. Of those women, significantly more Black (56.1%) than White (8.9%) women examined with the recommended frequency of more than 10 times a year. Of the Black women 60.5% and 8.4% of White women had never had a cervical smear test. Logistic regression analysis indicated that attitudes were an independent predictor for these two tests. PMID- 11729547 TI - Pain language of bone marrow transplantation patients. AB - Previous studies have shown that hypnosis may be effective in reducing intensity of pain among bone marrow transplantation patients whereas cognitive behavioral intervention without imagery was not effective for this group of patients. Since hypnosis alters patients' perception of pain and cognitive behavioral intervention changes patients' beliefs and improves their coping with pain, we hypothesized that sensory pain is more important than affective pain in understanding the pain experience of patients undergoing bone marrow transplantation. To test this hypothesis we administered the McGill Pain Questionnaire longitudinally to 50 consecutive eligible recipients of bone marrow transplantation during hospitalization to assess the different dimensions of pain they experienced. Consistent with our hypothesis, sensory pain fluctuated with treatment stages, and the pattern was consistent with previous findings. Patients reported significantly higher sensory pain than affective pain at all assessment points. In contrast, affective pain remained low and stable throughout the treatment. Our results contribute to the understanding of the nature of pain in bone marrow transplantation and suggest pain management strategies that focus on sensory pain as in hypnosis are more useful for such patients. PMID- 11729548 TI - Sibling dilution hypothesis: a regression surface analysis. AB - This study examined relationships between sibship size (the number of children in a family), birth order, and measures of academic performance, academic self concept, and educational aspirations at different levels of family educational resources. As part of a national longitudinal study of Australian secondary school students data were collected from 2,530 boys and 2,450 girls in Years 9 and 10. Regression surfaces were constructed from models that included terms to account for linear, interaction, and curvilinear associations among the variables. Analysis suggests the general propositions (a) family educational resources have significant associations with children's school-related outcomes at different levels of sibling variables, the relationships for girls being curvilinear, and (b) sibling variables continue to have small significant associations with affective and cognitive outcomes, after taking into account variations in family educational resources. That is, the investigation provides only partial support for the sibling dilution hypothesis. PMID- 11729549 TI - University students' conceptions of first-year mathematics. AB - The 1998 Crawford, Gordon, Nicholas, and Prosser Conceptions of Mathematics scale was administered to 156 first-year university students at a large public university in the midwestern United States. The scale represented fragmented and cohesive conceptions of mathematics. The reliability estimated as internal consistency had a Cronbach alpha of .80 for the fragmented scale and .87 for the cohesive scale. Factor analysis of the intercorrelations indicated the same two factors of fragmented and cohesive as in the original and other replicating studies. Students' conceptions of mathematics in this study were comparable to those reported in the original study. PMID- 11729550 TI - Clinical assessment of tics. AB - Comprehensive clinical evaluation of tics includes (1) assessments to diagnose (a) akathisia, myoclonus, stereotypies, and other dyskinesias and (b) their frequent behavioral concomitants, including attention deficits, compulsions, obsessions, and other psychopathologic manifestations, (2) descriptions of movement parameters from the patients, their parents, teachers, and important others, and (3) precise topographical localization of tics. PMID- 11729551 TI - Factors affecting behaviors that address HIV risk among senior secondary school pupils in South Africa. AB - The purpose of this study was to identify factors affecting HIV risk reduction among senior secondary school pupils in South Africa. The sample included 460 Grade 12, Secondary School pupils whose ages ranged from 16 to 30 years (M = 19.7 yr., SD = 2.5) and who were chosen at random from the total Grade 12 population throughout one region in the Northern Province of South Africa. Measures were of sexual behavior and condom use, knowledge about correct condom use, intention of condom use, behavioral norms, attitudes, normative beliefs, and subjective norms about condoms, HIV/AIDS vulnerability (likelihood to get it) and severity of the illness in the country, and condom use self-efficacy. Bivariate analysis gave positive significant relations among normative beliefs, subjective norms, and attitudes towards condom use as well as HIV/AIDS vulnerability and HIV risk behavior. Regression analysis indicated that for boys, younger age at first vaginal intercourse, less intention for condom use, and HIV/AIDS vulnerability were predictive for HIV/AIDS risk behavior and explained 39% of the variance. It is suggested that these predictors should be included in intervention programs for HIV prevention. PMID- 11729552 TI - Tolerance of ambiguity, information, and negotiation. AB - This study examines the relationship between the tolerance of ambiguity and the outcomes of negotiation and the misrepresentation of information. The data were obtained from 98 middle managers of a bank in Turkey. A positive correlation of .74 (p < .01) obtained between scores on tolerance of ambiguity with the outcomes of negotiation and a negative correlation of -.66 (p < .01) with misrepresentation of information. PMID- 11729553 TI - Mathematics anxiety among eighth-grade students of the United Arab Emirates. AB - To investigate the mathematics anxiety of 159 students in Grade 8 (M age = 14.3 yr.) in the United Arab Emirates a 28-item anxiety scale was administered. Analysis indicated some mathematics anxiety. Also, boys and girls scored significantly differently as boys had the higher mean. PMID- 11729554 TI - Temporal characteristics of slot machine play in recreational gamblers. AB - The present study examined the temporal characteristics of play exhibited by recreational slot machine players. 12 women (M = 24 yr., range 22-28) played a computerized version of a slot machine with probability of payoff, i.e., a winning spin, ranging from .2 to .6. Intertrial intervals or the time between the termination of one trial to the initiation of the next trial varied within subjects as a function of wins and losses. Winning trials were associated with larger intertrial intervals and losing trials with smaller intertrial intervals for 10 of the 12 women. No differences were found across probability levels between subjects. Results are discussed in terms of the post-reinforcement pause that occurs on specific schedules of reinforcement. PMID- 11729555 TI - Psychometric properties of the Illness Attitudes Scale in psychiatric patients. AB - This study assessed the factorial structure of the 29-item Illness Attitudes Scale in a population of 82 psychiatric patients hospitalized for gastrointestinal complaints. Factor analysis yielded three factors of the Illness Attitudes Scale in these patients, which have been interpreted Health Anxiety, Illness Behavior, and Health Habits. The internal consistency of these factors, estimated by Cronbach alpha, were .86, .80, and .58, respectively. The Health Anxiety and Illness Behavior scores were significantly intercorrelated .43, but scores on Health Habits were not significantly correlated with either Health Anxiety (r = .19) or Illness Behavior (r = .14). These findings are consistent with previous reports that two factors of the Illness Attitudes Scale possess reliable psychometric properties. Cross-validation with other patient populations is required to confirm the validity of the Illness Attitudes Scale factor structure testing at least 5 participants per item of the scale. PMID- 11729556 TI - Intergenerational association of alexithymic characteristics for college students and their mothers. AB - The intergenerational association of alexithymic characteristics of mothers and their children were examined in a sample of 232 pairs of college students and their mothers. Scores on the Toronto Alexithymia Scale, Parental Bonding Inventory, and the Family Environmental Scale of college students were significantly correlated with their mothers' memories of when they were also 20 years old. College students' scores were significantly correlated with their mothers' scores on each questionnaire. The student-mother pairs were further divided into two family types, nuclear and extended families. Correlations were higher for scores of the nuclear family than for those of the extended family. Such results suggest there may be intergenerational transmission of alexithymia and related factors from mothers to children. PMID- 11729557 TI - Reliability, validity, and factor structure of the Arabic version of the 12-item General Health Questionnaire. AB - The aim was to examine the psychometric properties of the Arabic 12-item General Health Questionnaire in a sample of university students. A sample of 157 university students was screened using this questionnaire and the Hopkins Symptom Checklist-90. A standardized clinical interview using SCID was conducted on a subset of screened students. Reliability, validity, and factor analysis of the questionnaire were evaluated. Using factor score discrimination between cases and noncases was also evaluated. The Arabic version of the GHQ-12 proved to be reliable as indicated by Cronbach alpha of .86. The best balance between sensitivity and specificity was found at the General Health Questionnaire cut-off point of 15/16: at this threshold, sensitivity was .88 and was paired with a specificity of .84. Principal component analysis with varimax rotation identified three factors, namely, Factor A (general dysphoria), Factor B (lack of enjoyment), and Factor C (social dysfunction). Factors A and C discriminated between clinically distressed and clinically nondistressed subjects. The General Health Questionnaire-12 as a whole is a reliable and valid screening tool in university settings. PMID- 11729558 TI - Alexithymia in schizophrenia: an exploratory study. AB - This study assessed a relationship between alexithymia and schizophrenia suggested by reports based on small samples of patients. Here, 50 outpatients with a diagnosis of paranoid schizophrenia were compared with 50 pair-matched healthy subjects. Alexithymia was measured by the Polish version of the Toronto Alexithymia Scale-20. The mean total score was significantly higher in the schizophrenia group (M = 52.3, SD = 13.47) than in the healthy controls (M = 45.8, SD = 11.39, p < .02). This replicates earlier findings showing that a group of patients with paranoid schizophrenia have higher scores on alexithymia scales than healthy controls. There seems a need for a comprehensive examination of relations between alexithymia and other concepts denoting pathology of affect in schizophrenia. PMID- 11729559 TI - Brief residential treatment for nicotine addiction: a five-year follow-up study. AB - Relapse is a major problem with nicotine addiction. This study investigated the long-term abstinence rate and effectiveness of a five-day residential nicotine treatment program. Questionnaires were mailed to all clients who completed the program between 1991 and 1996. 47% of the questionnaires were returned. Of these, 65% of respondents reported being abstinent from nicotine for an average period of 120 weeks. Of this group, 15% reported at least one relapse before a sustained abstinence. The primary relapse triggers for both the abstinent and nonabstinent groups were coping with "strong emotional feelings". Over 50% of the sample was comorbid for other addictions, depression or anxiety disorders. PMID- 11729560 TI - Law professor Stanley Herr dies; advocate won rights for disabled. PMID- 11729561 TI - Building community, recognizing dignity: beyond the ADA. PMID- 11729562 TI - "When I said that I was lying, I might have been lying": the phenomenon of psychological malingering. PMID- 11729563 TI - Malingering addendum. PMID- 11729564 TI - Interview with Charles S. Lauer, HFACHE, publisher, Modern Healthcare. Interview by Kyle L. Grazier. PMID- 11729565 TI - Creating a learning environment: a win-win approach. PMID- 11729566 TI - Impact of the "new science" of genomics. PMID- 11729567 TI - Suddenly out of the field, Part II: Dos and don'ts of moving on. PMID- 11729568 TI - Change in the population of health systems: from 1985 to 1998. AB - This article compares the predictions in Stephen Shortell's 1988 seminal article, The Evolution of Hospital Systems: Unfulfilled Promises and Self-Fulfilling Prophesies, with current data on health systems over a 14-year period from 1985 to 1998. Specifically, we review five of Shortell's predictions related to the horizontal growth of health systems and compare these predictions with empirical data on structural changes in the population of health systems. Our analyses suggest that Shortell's predictions corresponded to much of the actual behavior demonstrated in the population over the past one-and-a-half decades. Support was found for the following: (1) health systems form in two recurring stages; (2) previously unaffiliated hospitals are affiliating with existing systems rather than participating in the creation of new systems; and (3) health systems have evolved into five different strata, each of which represents different shares of the population; such population patterns have important implications for individual hospitals and health systems. By attending to patterns of change in the industry's social structure, hospitals and health systems can determine whether it is likely to continue along past trajectories or whether it shows signs of change that may pave way for the breakdown of existing organizational forms, entry of new organizational players, and the emergence of new governance structures. PMID- 11729569 TI - Race, ethnicity, and careers in healthcare management. AB - This article discusses the result of a study we conducted to investigate the factors that facilitate or impede healthcare management career opportunities and satisfaction from the perspective of racially and ethnically diverse healthcare managers. These healthcare managers were invited to participate in focus groups wherein they engaged in a structured discussion of six questions germane to race, ethnicity, and healthcare management careers. Nineteen themes were identified in the study analysis, eight of which emerged in all focus groups irrespective of race and ethnicity. In this article, we discuss the implications of the results for research and organization change and development and we recommend plans of action in four areas identified from the 19 themes: (1) industry leadership, (2) health administration education, (3) health management research, and (4) best practices. PMID- 11729570 TI - Automating the medication distribution process. PMID- 11729571 TI - Terror and medicine--the challenge. PMID- 11729572 TI - MEFV mutations and phenotype-genotype correlations in North African Jews and Armenians with familial Mediterranean fever. AB - BACKGROUND: Familial Mediterranean fever is a genetic disease in which some characteristic gene mutations have been found. OBJECTIVES: To analyze the phenotype-genotype correlations in North African Jews and Armenians with FMF. METHODS: We studied MEFV gene mutations and phenotype-genotype correlations in North African Jews and Armenians with Familial Mediterranean Fever living in France. RESULTS: M694V mutation was the most common mutation in Jews and in Armenians. Patients with M680I homozygosity or M680I/M694V compound heterozygosity had a phenotype as severe as patients with M694V homozygosity. CONCLUSIONS: This study characterizes the phenotype-genotype in specific ethnic groups of patients with FMF. PMID- 11729573 TI - Blood pressure values in children with intrauterine growth retardation. AB - BACKGROUND: Low birth weight has been shown to be strongly related to hypertension in adult life. OBJECTIVES: To determine whether blood pressure is higher in children with intrauterine growth retardation than in control subjects. METHODS: Blood pressure was measured in 58 children aged 4-6 years with IUGR and in 58 age-matched controls. The control children, whose birth weight was appropriate for gestational age, were also matched for gestational age. RESULTS: The children with IUGR had significantly higher mean values of systolic (P < 0.05) and diastolic blood pressures (P < 0.05) and mean arterial pressure (P < 0.05). Significant differences in blood pressure values were found between preterm IUGR (n = 21) and preterm controls (P < 0.05). CONCLUSIONS: These data indicate that children with IUGR may be at higher risk of hypertension already in childhood. PMID- 11729574 TI - The use of technologies to decrease peri-operative allogenic blood transfusion: results of practice variation in Israel. AB - BACKGROUND: Concern about the side effects of allogeneic blood transfusion has led to increased interest in methods of minimizing peri-operative transfusion. Technologies to minimize allogeneic transfusion include drugs such as aprotinin, desmopressin, tranexamic acid and erythropoietin, and techniques such as acute normovolemic hemodilution, cell salvage and autologous pre-donation. OBJECTIVE: To survey the current use in Israel of these seven technologies to minimize allogeneic blood transfusion. METHODS: Our survey was conducted in 1996-97 in all hospitals in Israel with more than 50 beds and at least one of the following departments: cardiac or vascular surgery, orthopedics, or urology. All departments surveyed were asked: a) whether the technologies were currently being used or not, b) the degree of use, and c) the factors influencing their use and non-use. The survey was targeted at the heads of these departments. RESULTS: Pharmaceuticals to reduce allogeneic blood transfusion were used in a much higher proportion in cardiac surgery departments than in the other three departments. Pre-operative blood donation was used in few of the cardiac, urologic and vascular surgery departments compared to its moderate use in orthopedic departments. The use of acute normovolemic hemodilution was reported in a majority of the cardiac departments only. Moderate use of cell salvage was reported in all departments except urology where it was not used at all. CONCLUSION: There is considerable practice variation in the use of technologies to minimize exposure to peri-operative allogeneic blood transfusion in Israel. PMID- 11729576 TI - Incidence of immunoglobulin G antibodies to Chlamydia pneumoniae in acute myocardial infarction patients. AB - BACKGROUND: Recent studies have suggested a possible association between Chlamydia pneumoniae infection and coronary heart disease. OBJECTIVES: To determine titers of antibodies to C. pneumoniae in patients with acute myocardial infarction compared with titers in several control groups. METHODS: This prospective case-control study investigated 209 individuals. We assessed the serum IgG antibody titers to C. pneumoniae in 57 consecutive patients admitted with AMI to our intensive coronary care unit during a 4 month period. A serum sample was drawn upon admission and after 6 weeks. Results were compared with those of four control groups: a) patients admitted with community-acquired pneumonia (n = 18), b) patients with community-acquired urinary tract infection (n = 42), c) patients with angiographically normal coronary artery disease (n = 44), and d) patients with stable coronary artery disease (n = 48). Serum immunoglobulin G antibody titers to C. pneumoniae were determined using standard micro-immunofluorescence technology. RESULTS: Of 57 patients with AMI, 32 (56%) had a high IgG titer to C. pneumoniae (> or = 1:256) on the initial test, which remained unchanged (62%) after 6 weeks. The percentage of patients with high titers was significantly lower in the control groups: 5 of 18 patients (28%) in the pneumonia group (P < 0.01), 11 of 42 (26%) in the urinary tract infection group (P < 0.01), 11 of 44 (25%) with normal coronary arteries (P < 0.01), and 17 of 48 (35%) with stable chronic ischemic heart disease (P < 0.05). CONCLUSION: The detection of high titers of IgG antibodies to C. pneumonia in many patients with AMI, compared to control groups, suggests that chronic Chlamydia pneumoniae infection plays a role in the pathogenesis of atherosclerosis and acute ischemic events. PMID- 11729575 TI - Fibroadenoma of the breast: analysis of associated pathological entities--a different risk marker in different age groups for concurrent breast cancer. AB - BACKGROUND: Fibroadenoma, one of the most common benign breast lesions, has a characteristic age-specific incidence and is associated with other pathological entities in 50% of cases. The clinical or imaging diagnosis of fibroadenoma may be erroneous, and in some cases is found to be invasive cancer. The clustering of such entities, their correlation with age, and the risk of synchronous breast malignancy are uncertain. OBJECTIVE: To explore the possibility of any significant clustering of fibroadenoma-associated benign breast diseases and to assess the possible risk of concomitant breast cancer. METHODS: We analyzed the pathological results of 147 women undergoing excisional biopsies for fibroadenoma diagnosed pre-operatively either by clinical examination and imaging (n = 17) or by radiology alone (n = 30). The inter-relationships among all entities associated with fibroadenoma were studied by hierarchical cluster analysis. The correlation of the various pathologies with the risk of invasive breast cancer in relation to the patient's age was also evaluated. RESULTS: Fibroadenoma associated pathologies were found in 48% of the cases: sclerosing adenosis (23%), duct ectasia (17.7%), apocrine metaplasia (15.6%), florid fibrocystic disease (12.9%), duct papillomatosis (11.6%), infiltrating duct carcinoma (5.4%), duct carcinoma in situ (3.4%), and 1 case of lobular carcinoma in situ (0.6%). An orderly internal hierarchy and three significant clusters emerged: a) epithelial apocrine metaplasia, duct ectasia and sclerosing adenosis (similarity coefficients 16.0, 11.0 and 8.0 respectively); b) papillomatosis, florid fibrocystic disease and calcifications (similarity coefficients of 6.0, 4.0 and 2.0 respectively); and c) infiltrating duct carcinoma and duct carcinoma in situ (similarity coefficients of 1.8 and 1.6 respectively). Seven of the eight patients with breast cancer were older than 40 years. CONCLUSIONS: In about half of the cases fibroadenoma was associated with other pathological entities clustered in an orderly hierarchy. The rarity of synchronous breast cancer in the younger age group and its more common association with fibroadenoma in the older age groups dictate a different approach to each. The finding of fibroadenoma in women older than 40 indicates the need for surgical excision. PMID- 11729577 TI - Medical care perceptions in elderly patients with musculoskeletal complaints. AB - BACKGROUND: Musculoskeletal complaints represent the second most common reason for visits to a physician, second only to the common cold. The limited capability of medical treatment for musculoskeletal disease requires modification of communication with patients by attending to their perception of the disease. OBJECTIVES: To assess patients' satisfaction with care provided by their primary physicians, and the relationship of patients' satisfaction to their expectations of that care, perceptions of physician performance, and perceived severity of musculoskeletal disease. METHODS: Questionnaires were administered to 90 community-dwelling elderly patients (mean age 76 +/- 8 years) presenting for follow-up appointments with their primary care physicians. Patients were asked to report on their satisfaction with the medical care provided by the primary physicians for musculoskeletal symptoms, their expectations of that care, their perceptions of their primary physicians' interaction (regarding competence, performance, and communication), and their perceptions of disease severity (based on the number of areas involved, pain frequency and intensity, and impact on daily activity). The effects on the degree of satisfaction were assessed with regard to demographic variables, co-morbidity, site involved, and response to recommended treatment. RESULTS: Most patients (> 85%) expressed overall satisfaction with their doctor's interpersonal skills. Fewer (76.9%) were satisfied with the amount of effort their doctors spend evaluating their musculoskeletal symptoms, the information received regarding their musculoskeletal symptoms (75%), the degree of pain relief (75%), and the degree of functional improvement (61.8%). Level of education and response to recommended treatment for musculoskeletal disease were the only parameters associated with degree of satisfaction (higher education P = 0.005, lower education P = 0.059, medication P = 0.008, rehabilitation P = 0.076). A high level of expectations (regarding physician's care and musculoskeletal disease treatment) was noted. CONCLUSIONS: The high level of patient satisfaction with their primary physicians' care for musculoskeletal symptoms may reflect the overall tendency of the elderly population to be satisfied with its primary care physicians. However, their high level of expectations (related to perceived efficacy of medical treatment) and their unrealistic perceptions of disease may lead to disappointment and non-compliance with their doctor's recommendations. Management of musculoskeletal disease in the elderly should address the patients' disease perceptions, as well as their therapeutic and functional needs. PMID- 11729578 TI - The role of trace elements in psoriatic patients undergoing balneotherapy with Dead Sea bath salt. AB - BACKGROUND: A beneficial effect was observed in patients with psoriasis vulgaris following balneotherapy with Dead Sea bath salt. OBJECTIVES: To evaluate the possible role of trace elements in the effectiveness of balneotherapy. METHODS: Serum levels of 11 trace elements were analyzed in 23 patients with psoriasis vulgaris who participated in a double-blind controlled study of balneotherapy with either Dead Sea bath salt (12 patients) or common salt (11 patients). Thirteen healthy volunteers served as controls. RESULTS: The mean pre-treatment serum levels of boron, cadmium, lithium and rubidium were significantly lower in patients compared to controls, whereas the mean pre-treatment serum level of manganese was significantly higher in patients compared to controls. Balneotherapy with Dead Sea bath salt resulted in a significant decrease (P = 0.0051) in the mean serum level of manganese from 0.10 +/- 0.05 mol/L to 0.05 +/- 0.02 mumol/L. The mean reduction in the serum level of manganese differed significantly (P = 0.002) between responders (% Psoriasis Area and Severity Index score reduction > or = 25) and non-responders (% PASI score reduction < 25). Following balneotherapy with Dead Sea bath salt the mean serum level of lithium decreased in responders by 0.01 +/- 0.02 mumol/L, whereas its level in non responders increased by 0.03 +/- 0.03 mumol/L. (P = 0.015). CONCLUSIONS: Manganese and lithium may play a role in the effectiveness of balneotherapy with Dead Sea bath salt for psoriasis. PMID- 11729579 TI - Epididymitis in childhood: a clinical retrospective study over 5 years. AB - BACKGROUND: Acute scrotal pain in children presents a major diagnostic and therapeutic challenge. Epididymitis has been considered uncommon in childhood. The clinical spectrum and therapeutic policy of the acute scrotum in children is continually being reassessed. OBJECTIVES: To determine whether there has been an increase in the incidence of epididymitis in children and to advocate a more selective surgical approach to the acute scrotum. METHODS: We conducted a retrospective review of 65 children admitted to our department of pediatric surgery with the diagnosis of acute scrotum during a 5 year period. RESULTS: Of the 65 children admitted with the diagnosis of acute scrotum, epididymitis was diagnosed in 42 (64.6%). The remaining cases included torsion of the testis in 12 patients (18.5%), torsion of the appendix testis in 5 (7.7%), scrotal pain and minimal physical findings in 4 (6.1%), and scrotal hematoma and idiopathic scrotal edema in one patient each. Doppler ultrasound of the groin, color Doppler ultrasound of the testis and testicular nuclide scintigraphy (Tc-99m scan) examinations were performed on 49, 30 and 57 occasions, respectively; the Tc-99m scan was the most effective tool. All the patients with epididymitis were diagnosed before surgical intervention and were treated conservatively. CONCLUSIONS: We observed an increasing frequency of epididymitis in children admitted with the diagnosis of acute scrotum. PMID- 11729580 TI - Primary pterygium recurrence time. AB - BACKGROUND: The time to recurrence after surgical removal of primary pterygium (pterygium) and the association between the rate of recurrence and the postoperative interval remain unclear. OBJECTIVES: To determine the amount of follow-up time needed to identify recurrence in patients after surgical removal of pterygium. METHODS: We reviewed the files of 143 patients (143 eyes) with recurrent pterygium to determine the interval from surgery to recurrence. RESULTS: Almost all (91.6%) of the recurrences appeared by 360 days after surgery. CONCLUSIONS: One year is the optimal follow-up time to identify recurrence of pterygium. PMID- 11729581 TI - Genotype-phenotype relation and correlation in familial mediterranean fever. PMID- 11729582 TI - The enigma of antigen selection and antigen recognition in systemic lupus erythematosus. PMID- 11729583 TI - Recombinant thyroid-stimulating hormone in differentiated thyroid cancer. AB - Recombinant TSH is effective in providing exogenous TSH stimulation for patients with differentiated thyroid cancer on thyroid hormone-suppressive therapy. It allows for detection of thyroid remnant and metastases by radioiodine scan and by serum thyroglobulin determination. The sensitivity and image quality of the WBS are similar after rTSH and after THSH withdrawal in the majority of patients. The equivalent 100% sensitivity of rTSH- and withdrawal-stimulated serum thyroglobulin measurement alone in identifying patients with radioiodine uptake outside the thyroid bed [38] may eventually lead to more extensive use of serum thyroglobulin testing after rTSH, with more selective application of radioiodine WBS [39]. Currently, a phase IV trial is in progress to evaluate the efficacy of rTSH-stimulated thyroglobulin levels as the primary modality for long-term follow up of low risk thyroid cancer patients. The use of rTSH prevents the morbidity, metabolic impairment and the risk of tumor progression associated with THST withdrawal, because of shorter exposure time to elevated TSH [38]. Furthermore, it decreases the radiation exposure of healthy tissues due to faster iodine clearance in euthyroidism. rTSH is well tolerated, with transient nausea in 10.5% and headache in 7.3% of patients. No antibodies specific to rTSH were documented, even after multiple courses of the drug. Currently, rTSH is suggested for patients who do not respond to hormone withdrawal or cannot tolerate hypothyroidism. For patients with low risk of tumor recurrence, rTSH-stimulated testing may be used at 6-12 months after postoperative I-131 ablation and with a repeat cycle of rTSH one year later, followed by testing every 3-5 years. In high risk patients, one set of negative I-131 scan and thyroglobulin test results after hormone withdrawal are recommended before using rTSH testing, because of a greater sensitivity of the withdrawal scan and because rTSH is not currently approved for subsequent I-131 therapy often indicated in these patients [24]. Subsequently, two cycles of rTSH testing are recommended at 6-12 month intervals, followed by testing every 1-3 years for at least the first decade after initial diagnosis. The cost of this commercially available form of rTSH has been considered a major impediment to its common use; however, this should be weighed against the loss of productivity of working hours related to withdrawal [40]. In the therapeutic setting, rTSH is the only acceptable option in a subgroup of patients with hypopituitarism, ischemic heart disease, a history of "myxedema madness," debilitation due to advanced disease, or inability to elicit TSH elevation due to continued production of thyroxine by thyroid remnant or metastatic tumor [33,38]. In conclusion, recombinant TSH facilitates the management of patients with differentiated thyroid carcinoma. It increases the sensitivity of thyroglobulin testing during thyroid hormone suppression therapy and enables radioiodine uptake for whole-body scan and occasionally for radioiodine therapy, without the need for prolonged THST withdrawal and its associated hypothyroidism, reduced quality of life and risk of tumor progression. PMID- 11729584 TI - Immune response to DNA in systemic lupus erythematosus. AB - Antibodies to DNA occur prominently in systemic lupus erythematosus and have been extensively studied as probes for underlying immune disturbances. These antibodies have features of DNA antigen drive. While previous models for this response posited DNA as simple and inert, recent studies have indicated that DNA is immunologically diverse and, depending upon sequence and backbone structure, can stimulate or suppress immune responses. In particular, bacterial DNA is immunologically potent and can function as both an adjuvant and immunogen, eliciting in normal individuals antibodies to sites exclusive to bacterial DNA. In mice genetically predisposed to autoimmunity, however, bacterial DNA can elicit anti-DNA autoantibodies under conditions in which mammalian DNA is inactive. These findings suggest that foreign DNA can serve as a trigger for anti DNA responses, with SLE reflecting a disturbance in antibody specificity and a shift from binding of sequential to backbone determinants. In contrast to bacterial DNA, mammalian DNA can suppress certain immune responses and prevent macrophage cytokine production. To the extent that self-DNA drives responses in SLE, anti-DNA production in this disease may reflect a failure of this suppression. The recognition of DNA's immune activities thus suggests novel possibilities for disease pathogenesis. PMID- 11729585 TI - Autoantibodies to ribosomal P proteins in systemic lupus erythematosus. PMID- 11729586 TI - Traumatic memory: a cause for postoperative delirium--a diagnostic dilemma. PMID- 11729587 TI - An unusual case of Dirofilaria conjunctivae infection suspected to be malignancy of the spermatic cord. PMID- 11729588 TI - Familial Mediterranean fever phenotype II in Greece. PMID- 11729589 TI - Hieronymus Bosch (1450-1516): paleopathology of the medieval disabled and its relation to the Bone and Joint Decade 2000-2010. AB - BACKGROUND: At the start of the Bone and Joint Decade 2000-2010, a paleopathologic study of the physically disabled may yield information and insight on the prevalence of crippling disorders and attitudes towards the afflicted in the past compared to today. OBJECTIVE: To analyze "The procession of the Cripples," a representative drawing of 31 disabled individuals by Hieronymus Bosch in 1500. METHODS: Three specialists--a rheumatologist, an orthopedic surgeon and a neurologist--analyzed each case by problem-solving means and clinical reasoning in order to formulate a consensus on the most likely diagnosis. RESULTS: This iconographic study of cripples in the sixteenth century reveals that the most common crippling disorder was not a neural form of leprosy, but rather that other disorders were also prevalent, such as congenital malformation, dry gangrene due to ergotism, post-traumatic amputations, infectious diseases (Pott's, syphilis), and even simulators. The drawings show characteristic coping patterns and different kinds of crutches and aids. CONCLUSION: A correct clinical diagnosis can be reached through the collaboration of a rheumatologist, an orthopedist and a neurologist. The Bone and Joint Decade Project, calling for attention and education with respect to musculoskeletal disorders, should reduce the impact and burden of crippling diseases worldwide through early clinical diagnosis and appropriate treatment. PMID- 11729590 TI - Perspectives. Revenues drop, health spending rises: what's a state to do? PMID- 11729591 TI - Plan of action. Former Quorum transitions to new GPO. PMID- 11729592 TI - Mapping a disaster plan. PMID- 11729593 TI - Image staffing woes. PMID- 11729594 TI - Hold the winning hand. Tips on negotiating to the satisfaction of all parties. PMID- 11729595 TI - Endoscopically speaking, how clean is yours? PMID- 11729596 TI - Addiction medicine for the primary care physician. AB - Addiction and the consequences including medical, social, and legal are seen almost daily in a primary care office. To effectively deal with these problems it is imperative that the physician view addiction as a disease. This article explores the disease concept and the basis for the view as well as providing a guide to diagnosis and assessment. Useful screening tools are also provided as well as a discussion of biochemical markers currently available. A guideline for delivering the diagnosis is also discussed. An overview of the stages of treatment is provided as well, with particular emphasis on what the primary care physician needs to know. Outcome data regarding types of treatment looking at abstinence, harm reduction, and economic impacts are provided. Finally a look at parity for addiction treatment with a discussion of the cost and the benefits to society are also outlined. PMID- 11729597 TI - Dementia: the University of Oklahoma autopsy experience. AB - The brain from 98 consecutive patients with the clinical diagnosis of dementia were examined at autopsy in a standardized fashion. Alzheimer's Disease was present in 79 of the cases, 76%, but represented the only diagnosis in 41%. Thus, almost 60% had another associated pathologic disorder. Cerebral amyloid angiopathy (CAA) represented the single largest subset, present in 25 cases. 40% were accompanied by either 1) small, microscopic infarcts or cortical scars, or 2) small collections of macrophages containing hemosiderin or small hemorrhages. CAA occurred with both atherosclerotic cortical infarcts and arteriolosclerotic subcortical pallor or lacunar infarcts. Alzheimer's Disease occurred with Diffuse Lewy Body (DLB) Disease in 13 cases. DLB Disease did not occur as a distinct entity, and thus may represent the second largest subset of Alzheimer's Disease. Both Alzheimer's Disease and DLB Disease accounted for dementia in Parkinson's Disease. Almost 25% of all cases had a disorder other than Alzheimer's Disease. PMID- 11729598 TI - The prevention of shaken baby syndrome. AB - Shaken Baby Syndrome may have life-long consequences, but it can be prevented. It is far too common and usually arises when the parent or caretaker gets angry, frustrated or impatient, but it can also be caused by inappropriate play. There are pathognomonic signs but suspicion must be high to suspect it. Physicians play an important role in diagnosis, management and most importantly, prevention. PMID- 11729599 TI - Health insurance shock may prompt changes. PMID- 11729600 TI - Premier databases to support FDA surveillance related to drug safety. PMID- 11729601 TI - Facility capacity constraints present a strategic planning opportunity. PMID- 11729602 TI - Hospital consultants see more interest in ER and other facilities expansion projects. PMID- 11729603 TI - Medical schools and hospitals adopt handheld personal data assistants. PMID- 11729604 TI - Harvard Medical School faculty provides credibility to Aetna-owned Web site. PMID- 11729605 TI - Hospitals' capitation reinsurance war exclusions will be toughened. PMID- 11729606 TI - Hospitals are more ready for bioterrorism. PMID- 11729607 TI - Humana reports 28.6% increase in earnings per share as medical cost rise 9%. PMID- 11729608 TI - [Tick-borne meningoencephalitis in Gorski Kotar--new findings]. AB - Motivated by the occurrence of a case of suspected tick-borne meningoencephalitis (TBE) in a forestry worker of the Delnice Forestry Enterprise, 139 sera samples were collected from his co-workers, for TBE-specific IgG antibody testing. The commercially purchased ELISA and our own indirect immunofluorescence tests were used. Two sera samples were found positive and another six as inconclusive. All of them were sent for confirmation to the Institute of virology, University of Vienna. The two previously positive sera were also ELISA positive in Vienna, the rest being qualified as negative. Accepting that the first case really had TBE and that the two seropositives have experienced infection with the TBE virus, it is inferred that in the wider region of Delnice there is an until now unknown TBE focus, quiescent there for an unknown period of time. PMID- 11729609 TI - [Importance of the CRH (corticotropin releasing hormone) test in the differential diagnosis of Cushing's syndrome]. AB - In the group of 13 patients with Cushing's syndrome (CS) CRH test was performed by sampling the blood from peripheral vein and in eight patients also after inferior petrosal sinus catheterization (IPSC) to resolve the disease etiology. In the group of patients with Cushing's disease (CD, n = 11), which was proven by surgery and adenoma immunohistochemistry, 10/11 had in CRH test the significant increase of cortisol and ACTH in the peripheral blood. Among two patients with ectopic ACTH syndrome one had the significant increase of both hormones in CRH test. After IPSC the ratio of ACTH in the petrosal sinus and in the peripheral vein was significant in 4/8 patients before, and in 6/8 after CRH administration. The intersinus gradient was significant in 3/8 patients before, and in 4/8 after CRH test. According to our results we can conclude that the determination of ACTH in the blood from peripheral veins after CRH administration is a very sensitive method for differential diagnosis of CS, while the results after IPSC were less sensitive in our conditions than those described in the literature. PMID- 11729610 TI - [Data analysis of daily reports in general medicine]. AB - Health priorities in a sample of 1,425 subjects of a defined urban population were studied from data collected routinely in general practice. Analysis of prevalence has shown 1/5 of circulatory diseases, 1/7 of respiratory diseases, 1/8 of musculoskeletal diseases, and 1/10 of mental disorders. According to incidence for the first medical examination, the leading diagnostic categories were diseases of the respiratory, circulatory, musculoskeletal, digestive, and genitourinary system. Both prevalence and incidence have shown that health priorities in the studied population were diseases of circulatory system, with hypertension, and diseases of the respiratory system, with common cold, of the highest prevalence. PMID- 11729611 TI - [Home artificial respiration in a girl with Ondine-Hirschsprung syndrome (Haddad's syndrome)]. AB - A case of 5.5-year-old girl with Haddad syndrome (congenital central hypoventilation syndrome-CCHS or Ondine's curse and Hirschsprung's disease) is presented. At the age of 1.5 months, due to severe colonic aganglionosis, temporary colostomy was performed. At the age of 4.5 years the sigmoid colon and left colon were resected and temporary colostomy was closed. Since birth, the sleep central hypoventilation characterized with shallow respirations and frequent apnoeas was noticed. The child's life was compromised and therefore the postoperative recovery was complicated and prolonged. Cardiac exam showed initial signs of pulmonary hypertension and right-sided failure. At the age of 4.5 years assisted positive pressure ventilation via face mask, with transportable ventilator was initiated during sleep and was successfully performed at home. Since then the cardiac exam showed no abnormalities. There are no significant abnormalities at neurological exam while EEG shows minimal irritative changes. The girl's physical and psychomotoric development is progressive and she is having an excellent peer contact. We expect even better quality of life after the implantation of the diaphragmatic pacemaker which is planned at early school age. PMID- 11729613 TI - [Pathogenesis of postoperative pain and treatment options]. AB - This article is a review of acute postoperative pain pathogenesis and treatment options. It is aimed at clinicians involved in postoperative pain management, for better understanding of the pain treatment strategy. Every procedure of antinociception should include modulation of different target points, because acute pain mechanisms and origins are different. There are three main intervention levels: peripheral, spinal and supraspinal. We should choose analgesics with two or more intervention levels simultaneously as well as combination of analgesics with different target points. Optimal analgesia with spinal opioids is achieved thanks to addition of supraspinal and peripheral analgesia components. It is considered that optimal acute pain treatment could prevent nerve system dysfunction and evolution of chronic pain syndromE, which is present in some of postoperative patients. PMID- 11729612 TI - [A bronchial polyp and foreign body in an adolescent]. AB - A fifteen-year-old adolescent with recurrent pneumonia in the last two years is presented. The polyp in the lobar bronchus for the right inferior lobe and a foreign body in posterobasal segmental bronchus were found on bronchoscopy. The polyp (patHohistologically confirmed granulomatous tissue) and foreign body (thorny fruit of small caltrops, Tribulus terrestris) were extracted through a rigid ventilatory bronchoscope. The polyp presumably developed from the thorn of the fruit stuck in bronchial mucosa. PMID- 11729614 TI - [Refractory arterial hypertension--where is the refractoriness? Hic rhodus, hic salta!]. AB - Arterial hypertension (AH) is the major independent risk factor for cardiovascular and cerebrovascular morbidity and mortality. Success in treatment of hypertonic patients is currently still far from acceptable in developing countries but also in the most of the developed countries in the world. There are numerous reasons and explanations for such poor therapeutic outcome, with poor patient's awareness of the need to pay attention to his own health and occasional superficial physician's approach being only some of them. However, AH treatment may present as an exceptional problem in some patients. Despite sufficient therapy duration and optimal doses of three types of antihypertensives, including one diuretic, no decrease in arterial blood pressure was observed in a low number of patients, so that such AH is called refractory (RH). RH may be primary if present since the beginning of treatment, or secondary if presenting after a period of successful treatment. In case of secondary RH, secondary forms of AH should be considered, like renovascular hypertension, primary aldosteronism, etc. In cases of primary RH, it is necessary to exclude pseudohypertension, pseudoresistance and pseudotolerance before characterizing a patient as really resistant, when the cause should be primarily sought in pathomechanisms of those organ systems which otherwise play an important role in AT homeostasis or in etiopathogenesis of AH, i.e. in kidney and heart. A significant reason for refractoriness may be drug interactions or applications of hypertensives which are not an optimal choice according to hemodynamic or hormone background of increased blood pressure. Clearly, it is not always easy to determine the background of refractoriness in practice, yet fortunately such patients are not frequent and additional efforts dedicated to them are not at the time needed for other patients. In this paper we presented a review of the most frequent causes of refractoriness in AH treatment, possible errors in diagnosing RH, and pathomechanisms responsible for the occurrence of resistance. Prognosis of RH is considerably worse than in otheR hypertonic patients and, since the possibilities of treatment are not extensive, a review of possible therapeutic approaches is also presented depending on the pathogenetic background of resistance. As RH patients have increased body mass index, and the success of the therapy was observed to depend on the salt intake, particular attention should be paid to instructions for patients on the change in lifestyle. The treatment of RH patients is a serious challenge which may unfortunately be frustrating and not infrequently have a fatal outcome due to either disease progression or patient's obstinacy. PMID- 11729615 TI - [Current management of patients with ventricular tachycardia]. AB - Ventricular arrhythmia has for decades been considered as a premonitory sign and risk marker of sudden death. Novel theories about arrhythmogenesis and conditions for the occurrence of sudden death, as well as evidence about proarrhythmic effect of antiarrhythmic drugs, have changed the views on the treatment of ventricular arrhythmia. Ventricular tachycardia (VT) is most often associated with structural heart disease: ischemic heart disease and previous myocardial infarction, cardiomyopathy (dilated and hypertrophic), arrhythmogenic right ventricular dysplasia, valvular heart disease (mitral valve prolapse), heart failure, condition after surgical correction of a congenital heart disease. Sometimes VT occurs without structural heart disease (congenital LQTS, Brugada syndrome, idiopathic VT). Today's standpoint is to treat only symptomatic and/or prognostically significant arrhythmias. Prognostic significance of VT mostly depends on the type and degree of structural heart disease and on global cardiac function. In patients with asymptomatic non-sustained VT and low risk for sudden death no treatment is needed or antiarrhythmics are administered. Conversely, in high risk patients implantation of automatic cardioverter-defibrillator is indicated. In the treatment of acute attack of VT the following can be used: electroconversion, cardiac pacing (overdrive), lidocaine, amiodarone, beta blockers, and occasionally magnesium or verapamil. In the prevention of recurrent arrhythmia and sudden death we can use: amiodarone, sotalol, mexiletin, phenytoin, beta-blockers, radiofrequency ablation, implantable cardioverter defibrillator, and in specific patients verapamil, pacemaker or left ganglion stellatum denervation. PMID- 11729616 TI - [The female athlete triad]. AB - This review offers some basic information on a syndrome described in 1992 as the female athlete triad. The increasing participation of women in competitive sports has led to significant accumulation of knowledge about potential pathological conditions due to strenuous exercise. Participation in sports that emphasize specific body image, psychological constitution of young female athletes and significantly lower daily calory intake cause the development of disordered eating, especially anorexia nervosa. Anorexia in combination with intensive training induces menstrual disorders, exercise-associated amenorrhea being the most important one. Low serum estrogen concentrations, as well as insufficient daily calcium intake have negative influence on bone mineral density, and the athletes have greater risk of developing osteoporosis and stress fractures. We described the diagnostic and therapeutic procedures necessary to detect and treat this syndrome. Education of physicians, female athletes and their coaches, as well as the screening during the annual examination, remain the most important measures of prevention. PMID- 11729617 TI - [Ethical considerations in pediatric intensive therapy]. AB - The author presents some ethical aspects of the decisions taken in pediatric intensive care units (PICU), and discusses the indications for the care of terminally-ill children. Assessment of the patient based on the clinical knowledge and experience of the pediatrician-intensivist must come first, well before the evaluation using various systems for scoring vital functions. The author discusses the ethical aspects of withholding and withdrawing treatment and resuscitation in the ICU. He maintains that the pediatric ICU must not be a place reserved for the dying and terminal patients. He stresses the role of the patient's relatives, and their need for due information and counselling. The importance of continuing education of the ICU staff is emphasized. PMID- 11729619 TI - Quality healthcare. PMID- 11729618 TI - [Euthanasia]. PMID- 11729620 TI - Developing network-based services in the NHS. AB - Networks, based upon informal relationships, have ensured that care was delivered to patients for many years. This informal organisation of care, based upon personal relationships, ensures that where the bureaucratic organisation fails the patient, health professionals' work together to network the resources the patient needs. Networks are not new. Formalising networks and recognising their potential to deliver seamless care is new. The NHS must ensure that networks are developed, allowing them freedom from bureaucracy to reach their potential. The Northern and Yorkshire Learning Alliance (NYLA) was established as part of the Northern and Yorkshire health community's efforts to radically improve care. The NYLA operates as a network with a small team of change experts working to develop change management and service improvement capacity across 10,000 square miles. As a network based organisation the team has learned many lessons, which may inform the development of clinical networks in England. PMID- 11729622 TI - Redesign of healthcare processes classification to improve the processes of gathering information and data processing on professional and organisational quality in healthcare. AB - One of the most important instruments for gathering information and processing data relating to professional and organisational quality in health systems is "healthcare processes classification". The authors found that a typical problem of many European countries is a lack of reliable information in the field of healthcare, mainly because the development of quality instruments, including healthcare processes classification, is not a priority for medical and other health professions. Additionally, it is difficult to update this instrument coherently with organisational changes and developmental achievements. This article describes the approach used by the University Medical Centre in Ljubjana, Slovenia, to redesign its healthcare processes classification in order to improve the quality of healthcare. PMID- 11729621 TI - Integrating knowledge workers and the organization: the role of IT. AB - Agency theory is primarily concerned with the relationship between the principal (employer/purchaser) and the agent (employee/contractor) in the issue of goal aligned behavior. Jensen and Meckling and others were not referring to a knowledge worker agent in their conceptualization of the principal/agent relationship. The significance of having a knowledge worker agent is that the decision rights are no longer located with the principal but with the agent. This in turn has a tremendous bearing on goal alignment and agency problems. We propose that information systems/information technology (IS/IT), in particular enterprise wide systems, can alleviate these agency problems. We illustrate this through a case example from health care, an industry with a high proportion of knowledge worker agents. PMID- 11729623 TI - An analysis of service quality at a student health center. AB - This study provides an analysis of the quality of service provided by a university health care center. Students' expectations and perceptions of the center's services are identified by a survey. A second survey was administered to the employees of the center to compare student perceptions to those of the employees. Based on the results of this study, specific recommendations are made to improve the performance of the center. The implementation of these recommendations should also reflect a change in the students' perceptions about the quality of service provided by the center. PMID- 11729624 TI - Evaluation of a program to improve the prescription-writing quality in hospital. AB - This paper aims to evaluate the effectiveness of a program designed to improve the quality of drug prescription-writing at a university hospital in France. Improvement actions included feed-back from yearly audits and the dissemination of recommendations on how best to write the prescriptions. A random sample of 30 stays was selected from among the hospitalizations for the year 1996. From each patient, medical records were searched for the first prescription order of the stay and its quality was assessed according to standards. A total of 872 records were relevant and included 3,289 medications. The results were compared to those obtained for the two previous years. Actions to sensitize prescribers resulted in an insufficient improvement of most indicators of prescription-writing quality with results remaining well below ideal standards. The hospital staff concerned had a positive opinion of the program which led to an awareness of prescription problems. This assessment showed that the program had a moderate impact on prescribers' practice and efforts must be continued. PMID- 11729625 TI - Pharmaceutical care management: a modern approach to providing seamless and integrated health care. AB - Institutional pharmaceutical services have widely evolved over the past 20-30 years. Hospital pharmacy practice has changed from a profession concerned chiefly with the bulk preparation and distribution of drug products to one centred on ensuring optimal drug therapy. Whereas hospital pharmacists were charged with maintaining large drug stock on nursing units, many of them now provide individualized patient therapies. The practice of hospital pharmacy has therefore become one encompassing all aspects of drug therapy, from the procurement of drugs and drug delivery devices, their preparation and distribution, to their most appropriate selection and use for each patient. Hospital pharmacy services have traditionally had little involvement at the key stages in patients' hospital care. This leads to the conclusion that the model of clinical pharmacy practice adopted by many pharmacy department hospitals is no longer appropriate for the demands of today's health-care services. Reviews many new models proposed for clinical pharmacy practice including an integrated model for providing a pharmaceutical care management approach in the health-care system. This model is a response to the failures of traditional drug therapy. It is primarily an idea about how health professionals and patient should integrate their work to obtain outcomes important to patients and clinicians. PMID- 11729626 TI - Managing costs and managing care. AB - With a defined population served, contracted provider panels and the nature of care delivery integration, managed care has provided a solution, though not a panacea, to provide equitable services, standardized and prevention oriented cares to its enrolled members. Combined with the earmarked capitation reimbursement system and a series of cost containment and utilization review techniques, managed care has also demonstrated potently its capacity in cost saving and quality promotion. Presents steps and measures related to managed care that federal government has taken to manage care and contain cost. It is crucial to identify and promulgate best practices continually, while managing utilization of resources for improving health care, containing cost, and equalizing medical care access to a greater proportion of the population. Concludes that it may take time for a universal adoption of managed care. However, Americans may actually benefit more from having a standard level of health care that managed care could achieve and provide. PMID- 11729627 TI - Pakistani and indigenous "white" women's views and the Donabedian-Maxwell grid: a consumer-focused template for assessing the quality of maternity care. AB - A criticism of using women's views as an assessment of the maternity services in the UK is that the views of some have been under-represented and inappropriately researched, in particular those from minority ethnic groups. In addition, there is criticism that "what" has been assessed within the maternity services, and other health-care arenas, has been driven by the perspective of those who provide the service rather than those who use it. The purpose here is to explore integrating Pakistani and indigenous "white" women's views with the Donabedian Maxwell grid, as a means of offering a consumer-focused template for underpinning quality assessment and quality assurance strategies for the maternity services in the UK. PMID- 11729628 TI - Evaluating evaluation: implications for assessing quality. AB - Aims to review the literature on evaluation and to examine research, theory and practice in relation to two key questions: What is evaluation and What is its contribution? Evaluation is increasingly a contemporary concern owing to growing demand for verification of programme results and the current political and economic climate. With the growth in evaluation activity in diverse fields, especially within new domains, those involved in health-care delivery can benefit from an overview of the universal fundamentals of research and theory that translate into improved practice. To evaluate effectively, there is a need for a full understanding of evaluation's nature, purposes and concepts. Identifies and reviews the key subdomains of evaluation, namely: definitions; theoretical underpinning; formulation of goals and objectives; specification of the programme; and cost-benefit analysis. PMID- 11729629 TI - Workforce planning. Breaking the bank. PMID- 11729630 TI - Workforce planning. Easy come easy go. PMID- 11729631 TI - Critical care. Only connect. AB - The establishment of a network for critical care services in five hospitals has led to a decrease in transfers of patients for non-clinical reasons. There have been no transfers outside the network's area. The introduction of common admission policies has led to more openness about bed availability. The introduction of the network has standardised data collection. The availability of extra funds and facilities was a big incentive to staff involvement. PMID- 11729632 TI - [Monoaminergic anorectic agents]. AB - Catecholamine, dopamine, serotonine (5HT) and neuronal histamine are anorectic monoamines and act as anorectic neurotransmitters. The anorectic agents as modulators of these monoamines inhibit appetite by activating release together with suppressing reuptake of those monoamines. The anorectic agents were classified in clinical use into either alpha 1, beta-adrenergic receptor agonists or 5HT-receptor agonist. Dexfenfluramine, a 5HT-receptor agonist, was inhibited in clinical use because of its cardiac complications including pulmonary hypertension and valvelar disease. Mazindol is an adrenergic agonist and the solitary anti-obesity drug used clinically in Japan. Sibutramine shows the effects of both beta-adrenergic and serotonergic receptor agonists. Sibutramine induces not only appetite suppression but also acceleration of peripheral energy expenditure. No histaminergic anorectics have been employed in the clinical situation to date. L-Histizine, precursor amino acid of endogenous neuronal histamine, is useful for suppression of food intake, lipolytic acceleration of peripheral adipose tissues and enhanced energy expenditure in both animals and humans. L-Histizine thus inspires development of more effective and safer anorectics that can be used without suffering from the rebound phenomenon of body weight. PMID- 11729633 TI - [Potential molecular targets for anti-obesity drugs--after the discovery of leptin]. AB - The discovery of the adipose-derived hormone leptin has generated interest in the interaction between peripheral signals and brain targets involved in the regulation of feedings and energy balance. Potential anti-obesity drugs can be based on any intervention between the neuropeptide and its receptor that would alter the biological responses mediated by the neuronal network, in particular, food intake, metabolism and energy expenditure. Modulation of neurons in the arcuate nucleus by leptin results in reduced expression of neuropeptide Y and agouti-related protein, and increased expression of pro-opiomelanocortin (the precursor of a-melanocyte-stimulating hormone) and cocaine- and amphetamine- regulated transcript. Whether leptin finds its way into general usage as an anti obesity drug, the use of modern methods to identify and target the components of leptin signaling pathway will form the basis for new pharmacological approaches to the treatment of obesity. PMID- 11729634 TI - [Beta 3-adrenergic receptor agonists--past, present and future]. AB - beta 3-Adrenergic receptors (beta 3-AR) play an important role in the thermogenesis in brown adipose tissue and lipolysis in white adipose tissue. beta 3-AR agonists developed in the early stages produced marked weight reduction and an anti-diabetic effect in rats and mice, but did not in humans, because of the difference in the chemical structure of the beta 3-AR. In 1995, a naturally occurring variant (Trp64Arg) of the human beta 3-AR gene was shown to be correlated with obesity and insulin resistance in Pima Indians. Moreover, the fact that white adipocytes produce various hormones and cytokines that cause life style-related disease was recently made clear. Because the reduction of the visceral fat is thought to be important to prevent these diseases, the expectations for the human beta 3-AR agonist having a novel anti-obesity effect are rising. Some interesting findings were recently reported with beta 3-AR agonists: the difference of the lipolysis was dependent on the existence of the Trp64Arg mutation and the up-regulation effect of the UCP1 and beta 3-ARs themselves in the adipose tissue and skeletal muscle. Therefore, we introduce informations (past, present and future) on beta 3-AR agonists in this paper. PMID- 11729635 TI - [PPAR gamma agonist and antagonist]. AB - Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a ligand activated transcription factor and functions as a heterodimer with a retinoid X receptor (RXR). Supraphysiological activation of PPAR gamma by thiazolidinediones can reduce insulin resistance and hyperglycemia in type 2 diabetes, but these drugs can also cause weight gain. Quite unexpectedly, a moderate reduction of PPAR gamma activity observed in heterozygous PPAR gamma-deficient mice or the Pro 12 Ala polymorphism in human PPAR gamma has been shown to prevent insulin resistance and obesity induced by a high-fat (HF) diet. We investigated whether functional antagonism toward PPAR gamma/RXR could be used to treat obesity and type 2 diabetes. We show herein that moderate reduction of PPAR gamma with an RXR antagonist or a PPAR gamma antagonist decreases triglyceride (TG) content in white adipose tissue, skeletal muscle and liver. These inhibitors potentiate leptin's effects and stimulated adiponectin levels, which increases fatty acid combustion and energy dissipation, thereby ameliorating HF diet-induced obesity and insulin resistance. Paradoxically, severe reduction of PPAR gamma by treatment of heterozygous PPAR gamma-deficient mice with an RXR antagonist or a PPAR gamma antagonist depletes white adipose tissue and markedly decreases leptin and adiponectin levels and energy dissipation, which increases TG content in skeletal muscle and the liver, thereby leading to the re-emergence of insulin resistance. Our data suggest that appropriate functional antagonism of PPAR gamma/RXR may be a logical approach to protection against obesity and related diseases such as type 2 diabetes. PMID- 11729636 TI - [Mitochondrial uncoupling protein as a target of pharmacotherapy for obesity]. AB - Uncoupling protein (UCP) is a transporter family present in the mitochondrial inner membrane, and as its name suggests, it uncouples respiration from ATP synthesis by dissipating the transmembrane proton gradient as heat. UCP is now recognized as a key molecule in metabolic thermogenesis such as cold- and diet induced heat production, which is a significant component of energy expenditure, and its dysfunction contributes to the development of obesity. Among the UCP family, UCP-1 is expressed exclusively in brown adipose tissue (BAT), while UCP-2 is present in many organs and UCP-3 is in skeletal muscle. BAT thermogenesis by UCP-1, which has been studied most extensively, is controlled directly by sympathetic nerves principally through the beta-adrenergic action of norepinephrine. Since the beta 3-adrenoceptor is present primarily in adipose tissues, its selective agonists stimulate BAT thermogenesis and also lipid mobilization in white adipose tissue without any noticeable effect on beta 1- and beta 2-adrenoceptos. Therefore, beta 3-adrenoceptor agonists would be promising for the pharmacotherapy of obesity. UCP gene expression is up regulated by ligands for nuclear receptors such as thyroid hormone receptor, peroxisome proliferator-activated receptors (PPAR) and retinoid-X receptor. Long chain fatty acids and some of their metabolites are known to activate PPAR and thereby lead to abundant expression of UCP, which may also contribute to increase in energy expenditure and prevention of obesity. The activity of UCP is suppressed by purine nucleotides but activated by fatty acids. Thus, fatty acids increase UCP mediated thermogenesis by direct activation of UCP and also by increased gene expression, implying some specific fatty acids or their derivatives as an effective anti-obesity tool. PMID- 11729637 TI - [Leptin-induced regulation of fat metabolism and its accumulation]. AB - Recent findings have shown that supplementation of leptin decreases body weight in leptin-deficient ob/ob mice through its suppressive effect on food intake and accelerating effect on energy expenditure, particularly on peripheral fat lipolysis. When endogenously hyperleptinemic obese rats were further induced to be hyperleptinemic exogeously using adenovirus vector, their body fat mass was reduced but not food intake. These findings implicate a direct lipolytic action of leptin on peripheral adipose tissues in obese rats because leptin transport capacity across the blood-brain barrier is almost saturated by the relative hyperleptinemia. Recovery from excessive body fat accumulation after adenovirus induced hyperleptinemia is much slower than that after caloric restriction because there may be difference between those treatments in decreased lipogenic enzymes activities and/or increased activities of fatty acid oxidative enzymes and thermogenic uncoupling proteins. The fat melting effects of leptin may show its crucial pharmacologic potencies to design therapeutic strategies against morbid obesity. The studies on leptin provide a better understanding for creative approaches to anti-obesity drug that are efficient for reducing body fat mass without harmful side-effects. PMID- 11729638 TI - [Inhibitors of absorption as anti-obesity drugs]. AB - There are two types of anti-obesity agents which are classified as inhibitors of absorption: inhibitors of lipid and carbohydrate absorption. Inhibitors of lipid absorption consist of lipase inhibitor (orlistat, Nomma Herb's extract (CT-II) and fat substitute (olestra, sucrose polyester). Orlistat is now available as an anti-obesity drug in the USA and Europe. CT-II may be useful as a functional diet. Application of fat substitute is still limited in snack food. As for inhibitors of carbohydrate absorption, alpha-glucosidase inhibitors are now available as anti-diabetic drugs. To develop these agents for anti-obesity drug, solution of adverse effects on the gastrointestinal tract are necessary. PMID- 11729639 TI - [Medicinal plant and its related metabolic modulators]. AB - We suggested a strategy for identifying anti-obesity drugs. The strategy aimed to prevent obesity through inhibiting intestinal absorption of dietary fat. Intestinal absorption of dietary fat was found to be reduced by tea saponin and chitosan through inhibiting pancreatic lipase activity, by chondroitin sulfate through inhibiting both pancreatic lipase activity and fatty acid absorption, and by lactosucrose through inhibiting beta-monoglyceride absorption. All these functional substances reduced plasma triglyceride levels previously elevated by oral administration of a lipid emulsion containing corn oil. Furthermore, these substances were found to cause reduction in perimetrial adipose tissue weight, which had been elevated by oral administration of a high fat diet (containing 40% of beef tallow) to female mice. Based on these results, a strategy for identifying anti-obesity drugs was discussed. PMID- 11729640 TI - Awareness of the new long-term care insurance system and social services for elderly care in non-medical junior college students. AB - The present study was conducted in order to investigate the awareness of the new long-term care insurance and social services for the elderly care among young people. Non-medical junior college students answered a self-administered questionnaire about the new long-term insurance and other related matters. The present study revealed that only a small percentage of students knew about "the new public long-term care insurance system" (13% for males and 11% for females), "care-manager" (11% and 8%), or "care-plan" (9% and 5%) in 1999, just one year before this insurance system took effect. In contrast, more than one third of the students knew about "home-help service" (41% and 47%), "long-term care institution for the elderly" (33% and 35%), and "elderly care nursing home" (33% and 36%). However, these rates were much lower than the rates among nursing students in 1998. An educational program for non-medical students should be recommended to help them to understand our rapidly aging society and to take an interest in public policies and social services for the elderly in Japan. PMID- 11729641 TI - Primary biliary cirrhosis associated with ulcerative colitis. AB - A rare case of concurrent primary biliary cirrhosis and ulcerative colitis is described in a 61 year-old Japanese male. Primary biliary cirrhosis was diagnosed on the basis of characteristic histologic findings and a positive serum mitochondrial antibody test. Ulcerative colitis was diagnosed from typical findings on colonoscopy and the histologic features of a sigmoid colon biopsy specimen. This is the 12th report of a patient presenting with the combination of primary biliary cirrhosis and ulcerative colitis. The potential autoimmune relationships on the basis of these conditions are discussed. PMID- 11729642 TI - [Vascular reconstruction in hepatobiliary surgery]. PMID- 11729643 TI - [Basic techniques in vascular surgery]. AB - The procedure for vascular resection and reconstruction includes exposure of the diseased vessel, temporary interruption of blood flow, resection of the diseased segment, vascular repair or anastomosis and restoration of blood flow. Adequate exposure, including the healthy portions on both the afferent and efferent sides of the diseased segment, facilitates subsequent steps in the procedure. Since the temporary interruption of blood flow could lead to ischemic and reperfusion injuries, it is necessary to take measures to protect organs that receive blood flow from diseased vessels. The basic principles in vascular suturing are single sutures through all layers and intima-to-intima approximation. Continuous simple sutures are often used for vascular repair and anastomoses. In the case of anastomoses between small arteries, however, interrupted simple sutures are preferred to prevent stenosis. Twisting, stenosis, and overstretch at the point of anastomosis are errors that result in failure of vascular reconstruction. The methods for overcoming size differences between anastomosed vessels should be understood. Topical heparinization is easy and a useful method to prevent the production of thrombus. Systemic administration of anticoagulants is not always needed. PMID- 11729644 TI - [Surgical technique for hepatic venous reconstruction in liver transplantation]. AB - In living-donor liver transplantation, one of the tactics of hepatic vein reconstruction is to obtain a long and wide venous orifice. The short hepatic vein should be reconstructed in the left liver graft with a caudate lobe. In the modified right liver graft, the tributaries of the middle hepatic veins should be reconstructed if there are no communicating veins between the middle and right hepatic veins. PMID- 11729645 TI - [Surgical techniques for vascular reconstruction of the portal vein and hepatic artery in living-donor liver transplantation]. AB - To decide how to reconstruct the portal vein and hepatic artery for liver transplantation, anatomical variation, diameter, length, and injury to vessels during surgery, and the quality of recipient vessels should be considered. Hence, it is of key importance for donor and recipient surgeries to prepare adequate vessels for reconstruction. For reconstruction of the portal vein, anastomosis with as large a diameter as possible is required to obtain good portal flow. In cases with sclerosing stenosis and old thrombus, technical innovations such as branch-patch, a conduit using a vein graft, and venoplasty using a venous patch are necessary. For reconstruction of the hepatic artery, selecting a satisfactory recipient artery, overcoming size mismatch, and gentle handling of a recipient artery with pathological changes are important. Arteries smaller than 3 mm are anastomosed with a surgical microscope using the united suture technique. The fishmouth technique or funnelization technique can be used for anastomoses with a significant size mismatch, and an autoarterial graft is used when arteries do not reach each other. PMID- 11729646 TI - [Indications for and techniques of hepatic vein reconstruction in hepatectomy for hepatocellular carcinoma]. AB - The indications for hepatic vein reconstruction (HVR) in resection of segments 7 and 8 for hepatocellular carcinoma (HCC) have been controversial. Although right hepatic vein ligation may not cause complications in some patients, hepatic parenchymal congestion and liver dysfunction occur in others. We performed HVR using autovein grafts in 6 HCC patients, patch grafts in 3, and direct anastomosis in 1, respectively. From this experience, we consider that HVR may be indicated in patients with a discolored hepatic area after hepatic vein ligation, and without inferior right hepatic vein and intrahepatic venous communication. Furthermore, although repeat hepatectomy has resulted in improved survival in patients with recurrent hepatic tumors, when the right hepatic vein has already been ligated, numerous intrahepatic communicating vein anastomoses develop. In repeat hepatectomy in such patients, uncontrollable massive bleeding from hepatic veins occurs. Therefore, HVR is recommended to maintain the previous hepatic vein anatomy. Recently, the indications for HVR have been studied to prevent hepatic vein congestion and liver dysfunction in both the remant and graft livers in living-related donor partial liver transplantation from the standpoint of hemodynamics. Additionally, the technique of direct hepatic vein anastomosis is presented. PMID- 11729647 TI - [Resection and reconstruction of the inferior vena cava for major hepatic resection]. AB - The inferior vena cava (IVC) is partially or segmentally resected in major hepatic resection for malignant hepatic tumors in case of possible direct invasion to the IVC wall or IVC tumor thrombosis. The reconstruction methods of the IVC are divided into three categories depending on the degree of IVC resection: simple suture; patch repair; and segmental replacement. In segmental replacement, a synthetic material such as a cylindrical expanded polytetrafluoroethylene (ePTFE) grafts is widely utilized as a substitute. The total hepatic vascular exclusion technique is usually necessary in concomitant resection of the suprahepatic IVC. When a longer duration of hepatic vascular exclusion is required to resect and reconstruct the suprahepatic IVC and hepatic vein confluence, in situ hypothermic perfusion, the ante situm technique, or ex vivo bench surgery must be applied. When an ePTFE graft is replaced in the resected IVC, a Carrel patch of the IVC is used for the hepatic vein orifice to maintain anastomotic patency. Alternatively, the hepatic vein can be reanastomosed to an inferior vena caval segment transpositioned from the intact infrahepatic IVC portion by replacing the resected infrahepatic IVC with an ePTFE graft. PMID- 11729648 TI - [Combined portal vein and liver resection for biliary cancer]. AB - Portal vein resection has become common in hepatobiliary resection for biliary cancer with curative intent. When cancer invasion of the portal vein is very limited, wedge resection followed by transverse closure is indicated. Longitudinal closure is contraindicated, as this procedure causes stenosis of the portal vein. In the case of right hepatectomy, segmental resection is feasible before liver transection. Reconstruction is completed with end-to-end anastomosis, in which an intraluminal technique is used for posterior anastomosis and an over-and-over suture for anterior anastomosis. More than 5-cm resection of the portal vein often requires reconstruction with an autovein graft. In the case of left hepatectomy, portal vein resection after liver transection is preferable. The resection and reconstruction method should be determined based on both the extent of cancer invasion of the right portal vein and the length of the right portal trunk. So far, we have aggressively carried out combined portal vein and liver resection in 106 patients with advanced biliary cancer (62 cholangiocarcinoma and 44 gallbladder carcinoma). Twenty-nine patients underwent wedge resections and 77 segmental resections (66 end-to-end anastomosis and 11 autovein grafting using an external iliac vein). In patients with hilar cholangiocarcinoma (n = 58), 3- and 5-year survival rates were 23% and 8%, respectively. Three patients survived for more than 5 years after resection. In contrast, the prognosis of patients with gallbladder cancer (n = 44) was dismal. All of the patients died within 3 years after surgery, although they survived statistically longer than unresected patients. These data suggest that portal vein resection has survival benefit for patients with cholangiocarcinoma. However, the indications for this procedure in gallbladder cancer should be reevaluated. PMID- 11729649 TI - [Indications for combined resection and reconstruction of the hepatic artery in biliary tract carcinoma]. AB - More than 10 years have passed since hepatic artery resection was first performed for the treatment of biliary tract cancer. The safety of this procedure has been established with the introduction of the microsurgery technique. However, the benefits of and indications for this treatment have not yet been clarified. Twenty-three patients underwent vascular resection (portal vein in 7, portal vein + hepatic artery in 9, hepatic artery in 7) among 114 resected patients with biliary tract cancer in our institution. The right hepatic artery was reconstructed by end-to-end anastomosis in most cases. The curative resection rate was 88.9% in hilar bile duct cancer. However, it was less than 50% in other carcinomas. Cumulative 5-year survival rates of vascular resection patients with hilar bile duct cancer, lower bile duct cancer, gallbladder cancer, and cholangiocarcinoma were 14.8%, 25%, 0%, and 0%, respectively. On the other hand, the rates were 38.9%, 0%, 0%, and 0%, in the stage III + IV patients who did not undergo vascular resection. The longest survival period among patients with hilar bile duct cancer and lower bile duct cancer was 85 months and 65 months, respectively, whereas it was 15 months in gallbladder cancer and 20 months in cholangiocarcinoma patients. No hilar bile duct cancer patient who survived for more than 3 years had lymph node metastasis. The longest surviving cholangiocarcinoma patient has received adjuvant chemotherapy consisting of 5 fluorouracil and cisplatin. It is concluded that patients with hilar bile duct cancer are good candidates for vascular resection. Adjuvant chemotherapy should be administered to gallbladder cancer and cholangiocarcinoma patients, because vascular resection alone does not result in prolongation of life in these patients. PMID- 11729650 TI - [En bloc resection of the hepatoduodenal ligament for advanced carcinoma of the biliary tract]. AB - Theoretically, en bloc resection of the hepatoduodenal ligament is considered to be the most radical procedure for advanced carcinoma of the biliary tract. However, this procedure involves perioperative difficulties such as hepatic ischemia, portal congestion, patency of reconstructed vessels, and high incidence of operative mortality, moreover, when it is combined with resection of the liver and/or pancreas. We developed several tactics in vascular reconstruction, hepatic resection, and pancreatoduodenectomy in order to decrease the operative morbidity and mortality. We preferred to anastomose the portal vein and hepatic artery separately to avoid total hepatic ischemia, and used the porto-systemic bypass during prolonged portal reconstruction. We resected the liver without vascular clamping to minimize hepatic ischemia, and employed the simplified method of pancreatojejunostomy. To date, we performed 4 hepatoligamentectomies and 4 hepatoligament-pancreatoduodenectomies with no operative mortality, although long term survivors were not encountered. This procedure should be evaluated with more clinical experiences after its safety and indication was established. PMID- 11729651 TI - [Autoimmune liver diseases and their borderline lesions from pathologic standpoint of view]. PMID- 11729652 TI - [Autoimmune liver disease and its variant]. PMID- 11729653 TI - [Alternations of p53 gene, microsatellite instability and proliferation associated antigen Ki-67 in the synchronous multiple colorectal cancers]. AB - To evaluate the malignant potential of synchronous multiple colorectal cancers, we studied clinicopathologically 31 synchronous multiple colorectal cancers resected at our hospital. We also compared the p53 gene mutation rate, replication error (RER) rate, and Ki-67 antigen positivity rate between these cancers and 90 sporadic colorectal cancers. There was no significant difference in lymphoid and venous invasion, hepatic metastasis, or stage of colon cancer between the two types of cancers. The p53 gene mutation rate was lower in synchronous multiple colorectal cancers (p < 0.05). The RER rate and positivity rate for Ki-67 antigen was significantly higher in these cancers (p < 0.05). These results suggest that some synchronous multiple colorectal cancers result from carcinogenesis in which RER genes are involved, as HNPCC does. In the patients with synchronous multiple colorectal cancers, it is clinically important to follow them carefully focusing on multiple metachronous colorectal cancers and multiple organ cancers. PMID- 11729654 TI - [Evaluation of endoscopic papillary balloon dilation (EPBD) using one-step catheter for removal of common bile duct stones]. AB - We compared the results of EPBD in 18 patients using one-step catheter (one-step group) with those in 46 patients using usual two-step catheter (two-step group). The frequency of endoscopy and bile duct clearance were the equal in both groups. Pancreatitis occurred in only one patient of two-step group. Elevations of the amylase values were the equal in both groups. Amylase values after EPBD tended to decrease in 11 patients whose amylase values were above 500 IU/l before EPBD. Using one-step catheter in EPBD was useful for removing common bile duct stones easily and safety. PMID- 11729655 TI - [A case of ulcerative colitis with Sweet's syndrome]. PMID- 11729656 TI - [A case of hepatocellular carcinoma with renal metastasis 1 year and 3 months after hepatectomy]. PMID- 11729657 TI - [A case of primary hepatic carcinoid tumor associated with gallbladder cancer]. PMID- 11729658 TI - [A case of acute posttransfusion hepatitis C of which we can estimate the occurrence by nucleic acid amplification test and cure after therapy with interferon]. PMID- 11729659 TI - [A case of pancreatic tuberculosis resembling pancreatic serouscystadenoma]. PMID- 11729660 TI - [A case of pancreatic ascites due to rupture of pancreatic pseudocyst successfully treated with continuous arterial infusion of nafamostat mesilate]. PMID- 11729662 TI - [How to modify the Japanese translation for rheumatoid arthritis]. PMID- 11729663 TI - [Expression of urokinase-type plasminogen activator (uPA) and uPA receptor (uPAR) plays a role in destruction of joint tissue in rapidly destructive coxarthropathy (RDC)]. AB - OBJECTIVE: Proliferation of small vessels including capillaries in joint tissues is one of the significant histopathological features of rapidly destructive coxarthropathy (RDC). To examine the relationship between the angiogenesis and bone destruction, expression of urokinase-type plasminogen activator (uPA) and uPA receptor (uPAR), which are necessary for angiogenesis was histochemically investigated. MATERIALS AND METHODS: Synovia and cartilage-bone tissues of femoral head were obtained from RDC patients at total hip replacement (THR). These joint tissues were fixed with 4% paraformaldehyde, and cartilage-bone tissues were decalcified, additionally. Immunohistochemical staining was performed on paraffin sections, using monoclonal antibodies against human uPA and uPAR. RESULT: There was a difference between expression of uPA and that of uPAR. Expression of uPA was observed in many types of cells such as osteoclasts, giant cells, fibroblast-like cells and macrophages, of which osteoclasts and giant cells were most prominent. On the other hand, expression of uPAR was detected mainly in fibroblast-like cells and macrophages and rarely seen in osteoclasts and giant cells. CONCLUSION: These results indicate that uPA-plasmin system contributes to bone destruction in RDC, and furthermore, that fibroblast-like cells and macrophages play an important role in activation of uPA-plasmin system. PMID- 11729664 TI - [A case of Behcet's disease with chronic and repeated sudden hearing loss: successful treatment with intravenous cyclophosphamide pulse therapy]. AB - A 54 year-old man who had been diagnosed as Behcet's disease since 1985 was admitted due to sypmtoms of fever, bilateral chronic hearing loss and repeated sudden left deafness after a partial laparoscopic gastrectomy for IIa type of early gastric cancer. Pure-tone audiometry, Bekesy audiometry, speech audiogram and auditory brain stem response revealed sensorineural hearing loss. The findings of magnetic resonance imaging, magnetic resonance angiography, single photon emission computed tomography and positron emission tomography ruled out a central nervous system disorder. The presence of an elevated C-reactive protein level, von Willebrand factor and plasmin alpha 2-plasmin inhibitor complex suggested vasculitis to be involved in the development of hearing loss. Although pulse-dose methylprednisolone therapy effectively arrested the acute progression of hearing loss, repeated audiograms showed that a modest dosage of oral prednisolone failed to maintain such improvement. However, after performing high dose cyclophosphamide (CY) therapy, a significant improvement in the hearing loss (more than 10 dB) was observed. As a result, CY is thus considered to be a potentially important treatment for sensorioneural hearing loss. PMID- 11729665 TI - [A case of primary Sjogren syndrome with repeated purpura]. AB - In Sjogren syndrome, purpura is one of its various well known eruptions. Although this disease state is assumed to be based on hypergammaglobulinemia, the details of its mechanism are unknown. We experienced a case involving a female patient with primary Sjogren syndrome showing repeated purpura on the legs, and examined her blood viscosity and histopathology. This girl developed Sjogren syndrome and was admitted to our hospital at 12-years-old. She underwent steroid treatment because of aggravation of the xerosis state and prominent purpura on the legs. Hypergammaglobulinemia was improved during the course; however, purpura appeared repeatedly. Although her blood viscosity was slightly higher than normal, this had no relation to purpura and serum gamma globulin values. Skin biopsy revealed necrotizing angiitis. These results suggest that the purpura of this case was caused not only by hyperviscosity from the hypergammaglobulinemia but also involvement of vasculitis by the primary disease. PMID- 11729666 TI - [A case of primary antiphospholipid syndrome complicated with pulmonary hypertension]. AB - We describe the case of 55-year old male with antiphospholipid syndrome (APS) who developed pulmonary hypertension without any thromboembolic episode. Multiple pulmonary perfusion defects suggestive of in situ thrombosis were observed. Hematological findings revealed microangiopathic hemolytic anemia and thrombocytopenia. These findings were improved by anticoagulant therapy. We monitored mean pressure of pulmonary artery (mPAP) and total pulmonary vascular resistance (TPR) before and after using vasodilator agents by Swan-Ganz catheter. mPAP and TPR showed improvement on treatment with oxygen supplementation therapy and Isosorbide administration. Previously 11 cases with APS complicated with pulmonary hypertension were reported. Majority of these patients have had recurrent venous thrombosis, particularly deep vein thrombosis often accompanied by pulmonary thromboembolism (8/11 cases, 72%). However in this case pulmonary hypertension with APS may be induced by in situ thrombosis in pulmonary micro vessels. PMID- 11729667 TI - [A case of polyarteritis nodosa limited to both calves with a low titer of MPO ANCA]. AB - A-57-year-old woman had suffered from high fever, general malaise, and loss of weight (6 kg) since January, 2000. She was admitted to our hospital in May, 2000, because of gait disturbance following to swelling of bilateral calves with severe pain beside the above symptoms. Laboratory data showed leukocytosis (WBC 10,000/microliter), high ESR (98 mm/hr), positive CRP (3.43 mg/dl), positive ANA (x160), high titer of RA (266 IU/l), and a low titer of MPO-ANCA (18 EU). T 2 weighted magnetic resonance image (MRI) showed intensity area of the M. gastrocnemius. Skin biopsy specimens of the right leg demonstrated necrotizing arteritis of small-sized arteries. These findings supported the diagnosis of limited type of polyarteritis nodosa (PN). In this report, we demonstrate a rare case of PN limited to both calves with characteristic MRI findings and a low titer of MPO-ANCA. PMID- 11729668 TI - [A case of chronic erosive polyarthritis which developed 14 years after an intestinal bypass operation with subsequent remission by intestinal revision]. AB - A 38-year-old female patient developed the symptoms and signs of arthritis in the right tarsal joint for the first time after 14 asymptomatic years following an ileo-colic intestinal bypass operation which had been performed as an emergency procedure for acute ileus due to intestinal adhesions caused by the previous abdominal surgery. Her arthritis took a progressively severe course thereafter, primarily involving the joints of both lower extremities, and 13 years after the onset of symptoms she still continued to have active polyarthritis. However, no concomitant skin lesions of any form had been recognized throughout the course of the arthritis. On the articular radiographs erosive changes were evident in the right tarsal joints and also in the MTP joints of both big toes. Laboratory examinations of the serum revealed negative results for rheumatoid factor, circulating immune-complexes, anti-nuclear antibodies, and anti-DNA antibodies, while the serum level of CRP as well as the erythrocyte sedimentation rate were elevated. Other routine laboratory tests were all unremarkable, and neither HLA-B 27 nor HLA-DR 4 were positive. Therapeutic drug regimens consisting of NSAIDs, oral as well as intraarticular steroids, DMARDs, methotrexate, and combinations of these drugs were unsuccessful in controlling the severe symptoms of the arthritis. In view of this, a revision operation of intestinal bypass was performed 13 years after the onset of the arthritis. After the revision the severe pain of the arthritis began to subside gradually, and 1 year and 6 months later the patient achieved complete remission of the arthritis, and her CRP and ESR values returned to normal. PMID- 11729669 TI - [Autoimmune diseases and Th 1/Th 2 balance]. PMID- 11729670 TI - [Possible implication of transcriptional coactivator in rheumatoid arthritis]. PMID- 11729671 TI - Distinct gene expression in the stomach following stress and alcohol exposure. AB - Two paradigms of acute stress in the rat were used to produce changes in the stomach. The first involved restraint stress combined with water immersion and the second utilized acute intragastric exposure to absolute ethanol. The mRNA expression of immediate early genes (IEG) such as c-fos, c-jun and NGFI-A, cyclooxygenase (COX)-2 and heat shock proteins (HSP) 70 in the stomach were studied using in situ hybridization histochemistry. Upregulation of IEG and HSP70 mRNAs were observed in the smooth muscle cells of muscularis mucosae, muscularis externa and blood vessels in response to water immersion-restraint stress or intragastric application of absolute ethanol. In the restraint stress model, IEG (c-fos and NGFI-A) mRNAs were induced in the pit and isthmus of the mucosa, while in the ethanol exposure model, IEG (c-fos, c-jun and NGFI-A) and HSP70 mRNAs were upregulated in the damaged epithelium, especially surrounding the deep erosions. COX-2 mRNA was detected in surface mucous cells under desquamation. These distinct gene expressions in the mucosa indicate that the two stress paradigms produce different cellular responses. These data provide new insights into cellular mechanisms that occur during the pathogenesis of acute gastric mucosal lesions. PMID- 11729672 TI - An investigation of the conus medullaris termination level during the period of fetal development to adulthood. AB - The spinal cord fills the length of the vertebral canal at the early period of intrauterine term. It is reported to extend to the level of the third lumbar vertebra at birth, because the vertebral column is growing more rapidly in the longitudinal direction than the spinal cord. The present investigation aimed to determine the changes in the termination level of conus medullaris (TLCM) from fetus to adulthood in a total of 285 individuals who had no defects in the central or peripheral nervous system, and were obtained from our Faculties of Medicine and Konya Maternity Hospital between 1992-1995. The age distribution was as follows: 36 fetuses, 20 prematures and 50 neonates, 51 children aged 1 to 7 years and 128 adults aged 15 to 68 years. In this study, for fetuses, prematures, neonates and children the TLCM was determined using ultrasonography. In addition, microdissection was used in fetuses to confirm the results obtained from the above technique. Also, magnetic resonance imaging was used in adults. During fetal life the end of the conus altered its levels from S5 to L3 vertebrae. The tip of the conus medullaris of the prematures and neonates ranged from L1 to L3 vertebrae. The tip of the conus medullaris in the children lay between the Th12 and L3 vertebrae, and in the adults it was between the Th12 and L2 vertebrae. There were slight differences between the prematures and neonates in terms of the TLCM. We concluded that there are differences in the TLCM between the age groups and therefore, especially in prematures and infants the determination of the tip of conus medullaris might be important for preventing postoperative neurological complications. PMID- 11729673 TI - Variations observed during a 1999 dissection course--their sum and analysis. AB - A systematic study of anomalies in all body areas of 12 male and 12 female cadavers was attempted from data obtained during a 1999 dissection course. A total of 229 anomalies (132 in males, 97 in females) were observed, and they were comprised of 142 varieties. Classified by system, there were 71 (45 varieties) muscular, 118 (71) vascular, 29 (18) neural, 10 (7) visceral and 1 bone anomaly; by region, there were 50 in the head and trunk, 56 in the upper limbs, and 52 in the lower limbs. The other 71 anomalies were vascular ones confined to the viscera (61) and visceral (10). A maximum of 26 anomalies, including 10 muscular, 14 vascular and 2 neural anomalies, were ascertained to exist coincidentally in a single male. Among vascular anomalies, the visceral ones (61) were dominant. In all structures, 75% of the varieties of anomalies were single occurrences. In the text, the multiple occurrence of anomalies in an individual and their relationships are discussed. PMID- 11729674 TI - [Ultrastructural studies of hepatoblast junctions and liver hematopoiesis of the mouse embryo]. AB - To clarify the morphological changes in hepatoblast connections during the development of fetal liver hematopoiesis, ICR mouse livers of 11 to 19 days of gestation were studied by means of three-dimensional reconstruction, immunohistochemistry, electron microscopy and freeze fracture replica method. Embryonic liver weight showed rapid increase until 19 days of gestation, and an initial steep increase, due to hematopoietic development, was observed at 13 to 15 days of gestation. Hepatoblast volume appeared to be constant until 13 days of gestation, and, thereafter, showed a gradual increase. An 11-day primitive hepatic cord contained a few immature hematopoietic cells among hepatoblasts, and the hepatoblasts made contact with one another by short cytoplasmic projections. The area of the contact surface had a diameter of 4-5 microns, where E-cadherin mediated adherens junctions were found. At 12-13 days of gestation, hepatoblasts surrounded large ellipsoidal hematopoietic foci, with long cytoplasmic projections. In addition to the adherens junctions, small desmosomes appeared to bind hepatoblasts together, and biliary canaliculi could be recognized between hepatoblasts. At peak stage of liver hematopoiesis at 14 days of gestation, both tight junctions and gap junctions appeared around the biliary canaliculi, and four types of specialized junctions, i.e., adherens junctions, desmosomes, tight junctions and gap junctions, appeared to be fully developed. After 15 days of gestation, hepatocyte volume showed rapid increase, and the surface areas between adjacent hepatocytes were markedly enlarged. As a result, the involuted hematopoietic foci were forced to move from interhepatocytic spaces to perisinusoidal space at the end of the intrauterine life. PMID- 11729675 TI - [Alterations in neuroepithelial intermediate filaments during neurogenesis in the chick cervical spinal cord]. AB - Intermediate filament proteins including nestin, vimentin and neurofilament were immunohistochemically studied during neurogenesis in the chick cervical spinal cord from stages 8 to 28. At stage 8, neuroepithelial cells of the neural groove contained a large amount of nestin in their cytoplasm and a little vimentin in the basal cytoplasmic areas, and no neurofilaments could be recognized at all. At stage 10, there was a marked decrease in nestin expression in the neural groove, and there was an increase in vimentin in neuroepithelial cells. At stage 15, when the neural tube was formed, small oval neuroblasts appeared in the peripheral area of the neuroepithelium. By employing double-immunostaining, three different neuroblasts could be identified; vimentin-positive and neurofilament-negative cells, neurofilament- and vimentin-double-positive cells, and neurofilament positive and vimentin-negative cells. During the neuroblast stage, intracellular intermediate filaments were relayed from vimentin to neurofilaments. At stage 20, large polygonal cells containing a large number of neurofilaments could be recognized in the enlarged basal plate of the neural tube. At stage 28, neuronal processes developed in large polygonal cells and, although the staining intensity of the neurofilaments was slightly decreased in the soma, the neuronal processes contained a large number of neurofilaments. During neurogenesis in the chick cervical spinal cord, the intermediate filaments, nestin and vimentin, are present in neuroepithelial cells. During the neuroblast stage, vimentin and neurofilaments are observed together for a short time. Finally, in polygonal neurons, only neurofilaments are observed. PMID- 11729676 TI - [Anatomic and radiologic considerations of varied appearances of thoracic structures]. AB - The purpose of this pictorial essay was to demonstrate normal chest anatomy and related pathologies on chest radiographs and chest CT images. It is important for the general practitioner to have a clear understanding of anatomy in order to avoid overestimating subtle radiologic findings and to be able to differentiate true pathological lesions. This pictorial essay includes various appearances of pleural fissures, companion shadows of the ribs, and minor structures of the chest walls and mediastinum. PMID- 11729677 TI - [Two-dimensional and three-dimensional CT diagnosis of alimentary tract]. AB - The recent development of multidetector-row CT(MDCT) has made it possible to obtain three-dimensional images of the alimentary tract that offer new diagnostic potential. In its two-dimensional diagnosis of the alimentary tract, MDCT has also changed the concept of the oral contrast agent. Before MDCT, we routinely used a positive contrast agent to distinguish the stomach and intestine from other organs and masses. The excellent slice profile acquired by MDCT can distinguish the alimentary tract and depict abnormal findings without the use of a positive contrast agent. With the use of an intravenous contrast medium, the alimentary tract itself, alimentary tumors, and inflammatory disease are well demarcated with water and air. Moreover, the combination of two-dimensional and three-dimensional diagnostic images makes it possible to detect and assess early gastric and colonic cancers as conventional gastroscopy and colonoscopy. Although the lack of texture information is one of the disadvantages of three-dimensional CT, three-dimensional CT diagnosis of the alimentary tract is less invasive and more objective than conventional studies. Advances in three-dimensional imaging with isotropic data sets will lead to the use of two-dimensional and three dimensional CT diagnosis as one of the standard examinations of the alimentary tract. PMID- 11729678 TI - Development of an expert system for automatic chart checking combined with departmental digital chart system. PMID- 11729679 TI - [Preliminary experience of wireless teleradiology system using Personal Handyphone System]. AB - We investigated the potential of the wireless teleradiology system, using a Personal Handyphone System(PHS) and hand-held personal computer. To provide and interpret CT images, an internet web system was used. It took 5.5 seconds to transmit a single CT image of 28 KB. Therefore, about 2 minutes was required to brawse a series of brain examinations(20 CT images). Although transmission speed needs to be increased, our system makes it possible to perform imaging diagnosis anywhere and is especially useful in emergency situations. PMID- 11729680 TI - [Telangiectatic focal nodular hyperplasia: a case report]. AB - Focal nodular hyperplasia(FNH) is a benign tumor-like lesion of the liver. The telangiectatic type is an atypical histologic variant of FNH, and descriptions of its radiologic findings are rare. We report a case of telangiectatic FNH. A 26-20 mm round lesion was found in the lateral segment of the liver. Although this lesion appeared similar to cavernous hemangioma with rapid intratumoral contrast material enhancement on cross-sectional images, its signal intensity decreased on T2-weighted imaging after the administration of superparamagnetic iron oxide(SPIO), and its appearance differed from that of cavernous hemangioma. PMID- 11729681 TI - [Development of a guide wire with multiple markers to aid in port-catheter system placement]. AB - We developed a guide wire with multiple markers for use during implantation of the port-catheter system for intraarterial infusion chemotherapy. With the markers on this guide wire, the distance from the common hepatic artery to the right gastroepiploic artery can be precisely measured. From this measurement, it is possible to determine the distance from the side hole to the catheter tip; unneeded catheter can be cut off with surgical scissors. In all eight cases in which this guide wire was used, we implanted the port-catheter system in the ideal position. PMID- 11729682 TI - [Social conditions in the daily life of asthmatic patients]. AB - To investigate the social conditions in the daily lives of asthmatic patients in Japan, a nationwide survey was performed using a questionnaire compiled by a joint research group and conducted by The national hospital treatment joint research group and The national sanatorium central research group. This study was carried out on 3,331 patients with childhood asthma and 4,398 patients with adulthood asthma in 1995. The results were as follows. (1) More than half of the asthmatic patients had been admitted to hospital at least once. (2) When experiencing an asthmatic attack, more than 80% of patients had difficulties with daily activities. Even when not experiencing an asthmatic attack, 14% of patients with childhood asthma and 40% of patients with adulthood asthma had difficulties with daily activities. The frequency of difficulties in daily life increased when the asthma was more severe or the patient was elderly. (3) More than 60% of adulthood patients felt that there were obstacles to their hopes or plans in life. The frequency of obstacles increased in patients with more severe asthma and in more elderly patients. (4) Fifteen percent of adult patients experienced work disabilities due to asthma (giving up work, changing job, leaving school, changing their school). (5) Because of asthmatic attacks, 64% of child patients and 49% of adult patients needed to take at least one day off. The outcome of this survey highlighted the condition of asthmatic patients in Japan. During the treatment of asthma, not only control of asthmatic attacks but also the social conditions of the patients in their daily lives should be considered and addressed. PMID- 11729683 TI - [The impact of obesity on oxygen desaturation in patients with sleep apnea/hypopnea syndrome]. AB - We performed a cross-sectional investigation on the impact of obesity on clinical manifestations in patients with obstructive sleep apnea/hypopnea syndrome (SAHS). The subjects were 87 patients who underwent overnight polysomnography with an apnea/hypopnea index (AHI) of 15/h or more. We divided these patients into non obese (N) and obese (O) groups based on the median value of the body mass index (BMI), 27 kg/m2. Subjective symptoms, sleep quality and AHI were similar in both groups, but every parameter related to oxygen desaturation was worse in group O than in group N. There was no difference in the mean duration of apnea events between the two groups. The rate of fall in oxygen saturation during apnea events was highly correlated to the BMI (r = 0.72; p < 0.00001). Accordingly, we concluded that profound desaturation in group O is due to a rapid fall in oxygen saturation during apnea events compared with group N. Anthropometric measurements revealed that the rate of fall in oxygen saturation was more related to abdominal circumference (AC) than the neck circumference (NC), which is contrasted with the finding that AHI was more related to NC than AC. This fact suggests that abdominal obesity may deteriorate oxygenation during apnea events and may therefore aggravate the risk of cardiovascular disease in patients with SAHS. PMID- 11729684 TI - [Evaluation of spontaneous pneumothorax in patients with pulmonary lymphangioleiomyomatosis]. AB - We retrospectively evaluated the development of spontaneous pneumothorax in patients with pulmonary lymphangioleiomyomatosis (LAM). Fifteen patients with LAM, whose diagnosis was confirmed histopathologically, were enrolled. All were women, and had a mean age at diagnosis of 31.6 +/- 7.3 years. They were divided into two groups according to the presence or absence of pneumothorax at the time of onset, Group A consisting of nine patients with pneumothorax, and Group B, of six without pneumothorax. Spontaneous pneumothorax developed in 15 out of a total of 18 hemithoraces in Group A patients in whom 13 hemithoraces were surgically treated for pneumothorax (eight open thoracotomies and five video-assisted thoracoscopic surgeries). Bullectomy with either parietal pleurectomy, pleural abrasion, or instillation of chemical irritants to prevent the recurrence of pneumothorax offered better outcomes than bullectomy alone in terms of the postoperative recurrence rate of pneumothorax (p < 0.05). On the other hand, three of the five Group B cases developed pneumothorax during the course of LAM, but none was operated because of severely impaired lung function. The other patient in Group B was not only free of pneumothorax at onset, but also did not develop pneumothorax or suffer from impaired lung function. There were no deaths due to pneumothorax recorded among Group A patients, but two patients in group B died. A better survival rate was noted in Group A than in Group B by Kaplan-Meier analysis, suggesting that these two groups may represent the broad clinical spectrum of LAM. PMID- 11729685 TI - [Drug-induced pneumonitis caused by sulfamethoxazole-trimethoprim]. AB - A 33-year-old man was treated with sulfamethoxazole-trimethoprim (SMT-TMP) for an infection in the cervical vertebrae by methicillin-resistant Staphylococcus aureus (MRSA). Two weeks later a fever of 39 degrees C appeared, and a productive cough, hemosputum, and dyspnea developed a further three weeks later. Chest radiographs showed bilateral ground-glass opacity. Cell differentiation of bronchoalveolar lavage fluid (BALF) revealed increases of lymphocytes and eosinophils, and the CD4/CD8 ratio of the BALF lymphocytes was decreased. A thoracoscopic lung biopsy specimen showed fibroedematous thickening of the alveolar walls, hypertrophic alveolar cells, and cell infiltration with neutrophils and lymphocytes. The pathological diagnosis was nonspecific interstitial pneumonia, group II. The fever resolved 6 days after discontinuance of SMX-TMP. The lymphocyte stimulation test for SMX-TMP gave a positive result. Administration of glucocorticoid improved both the symptoms and the laboratory data. PMID- 11729686 TI - [A case of pleural effusion in a patient with malignant lymphoma diagnosed by thoracoscopy with local anesthesia]. AB - A 47-year-old woman was admitted for examination of massive right pleural effusion on chest radiography, together with chief complaints of dry cough and body weight loss. Exudative effusion was aspirated by thoracentesis, revealing many lymphocytes and a high level of adenosine deaminase. Thoracoscopy with local anesthesia was performed. Elevated white lesions and miliary nodules were observed, and these lesions were examined by thoracoscopic biopsy, leading to a diagnosis of malignant lymphoma. Treatment with chemotherapy (THP-COP) resulted in partial remission. Thoracoscopy with local anesthesia in cases of pleural effusion with malignant lymphoma can be a safe and useful procedure for diagnosis. PMID- 11729687 TI - [Invasive pulmonary aspergillosis following influenza A infection]. AB - A 55-year-old man with arc welder's pneumoconiosis who presented with bilateral pneumonia was admitted to our hospital. Streptococcus pneumoniae and Aspergillus fumigatus were cultured from sputum on admission. The patient was treated with antibiotics and an anti-fungal agent, but his chest radiograph shadows became exacerbated, and he died of respiratory failure on the fourth day of hospitalization. Histological examination of postmortem lung tissue revealed necrotizing aspergillous pneumonia. The results of hemagglutination inhibition tests for influenza A (H3 N2) were x16 in September 1999, and x512 on the third day of hospitalization. We diagnosed this patient's condition as invasive pulmonary aspergillosis associated with influenza A viral infection. The suppression of cellular immunity, lymphocytopenia, and destruction of airways mucociliary transport induced by influenza A viral infection were suspected to have predisposed him to aspergillus superinfection. PMID- 11729688 TI - [A case of Mycobacterium intracellulare infection complicated by immotile cilia syndrome]. AB - A 25-year-old woman with a history of immotile cilia syndrome (ICS) was admitted to our hospital with dyspnea. Chest roentgenography revealed dense infiltrates in both lower lung fields in addition to bronchiectasis and small nodular opacities, which had been observed previously. Transbronchial lung biopsy demonstrated evidence of non-caseating epithelioid cell granuloma. Sputum specimens were examined, and isolates were identified as Mycobacterium intracellulare. The patient was given antituberculous therapy and clarithromycin, which induced clinical improvement. It is well known that bronchial mucociliary transport is severely impaired in patients with ICS. However, to our knowledge, cases of M. intracellulare infection complicated by ICS have not been reported in Japan. We must pay close attention to the concurrence of these diseases. PMID- 11729689 TI - [Occurrence of BOOP outside radiation field after radiation therapy for small cell lung cancer]. AB - We report a case of bronchiolitis obliterans organizing pneumonia (BOOP) that occurred outside the radiation field after radiation therapy for small cell lung cancer. A 74-year-old woman received chemotherapy and a total of 60 Gy of radiation therapy to the right hilum and mediastinum for small cell carcinoma of the suprahilar area of the right lung. Radiation pneumonitis developed within the radiation port 3 months after the completion of radiation therapy. She complained of cough and was admitted 7 months after completion of the radiation therapy. Chest radiography and computed tomography demonstrated peripheral alveolar opacities outside the radiation field on the side contralateral to that receiving the radiation therapy. Bronchoalveolar lavage showed that the total cell count was increased, with a markedly increased percentage of lymphocytes. Transbronchial lung biopsy revealed a histologic pattern consistent with BOOP. Treatment with corticosteroids resulted in rapid improvement of the symptoms and complete resolution of the radiographic abnormalities of the left lung. Although some cases of BOOP following radiation therapy for breast cancer have been reported, none of BOOP after radiation therapy for lung cancer have appeared in the literature. PMID- 11729690 TI - [A case of pulmonary actinomycosis with recurrent hemoptysis diagnosed after right upper lobectomy]. AB - The patient was a 58-year-old man who had been admitted in 1995 because of hemoptysis. Chest CT scans showed air-space consolidation with dilated bronchi and calcification in the right S3. He received a diagnosis of bronchiectasis with old tuberculosis. Bronchial arteriography showed arterialization in the right S3, and bronchial artery embolization was performed. But in 1996 hemoptysis reappeared. He was readmitted in May 1999 because of recurrent hemoptysis. Bronchial arteriography showed recurrence of arterialization in the same area, and chest CT scans showed growth of the mass shadow. Right upper lobectomy was performed, and the microscopic findings of the resected specimen showed sulfur granules in the dilated bronchus. We concluded that pulmonary actinomycosis should be considered in the differential diagnosis of abnormal chest shadows. PMID- 11729691 TI - [Two cases of alveolar hemorrhage due to inhalation of detergent aerosol]. AB - We report on two cases of alveolar hemorrhage after inhalation of detergent aerosol. Case 1: A 54-year-old woman was referred to our hospital with a complaint of hemoptysis. She used a spray type detergent for ventilation fan cleaning, and recognized hemoptysis later for eight hours. She had mild anemia and severe hypoxemia, and chest radiographs and CT films infiltrative shadows were recognized predominantly in the right middle, left lingual lower lobes. Case 2: A 22-year-old woman was admitted to our hospital complaining of bloody sputum and dyspnea. She had used a spray-type detergent for bathroom cleaning intermittently for one week. Between the beginning of this activity and the seventh day, cough and dyspnea developed. Chest radiographs and CT films disclosed diffuse infiltrative shadows in both lung fields. In both cases, the condition was diagnosed by bronchoalveolar lavage as alveolar hemorrhage. In case 2, a transbronchial lung biopsy specimen revealed alveolitis without vasculitis or capillaritis. Both the clinical symptoms and the CT images were improved by steroid administration. The steroid dosage was decreased gradually and stopped, but there was no relapse of the hemoptysis. Neither patient was positive for antinuclear antibodies or antibodies against the cytoplasm of granulocytes. Urinalysis showed no pathological findings. Since the hemoptysis appeared after inhalation of a detergent aerosol, it was possible that inhalation was the cause of the hemorrhage. PMID- 11729692 TI - [A case of endobronchial tuberculosis associated with bronchial asthma treated with high doses of inhaled corticosteroid]. AB - A 46-year-old woman had been treated with 1,600-2,000 micrograms/day of beclomethasone dipropionate (BDP) and oral theophylline on the basis of a diagnosis of bronchial asthma in 1993. Eosinophilic pneumonia was diagnosed in June 1999, and she was then treated with 40 mg/day of oral prednisolone (PSL), which was gradually tapered off, and then stopped in October 1999. She was referred to our hospital because acid-fast bacilli were found in the sputum on January 18, 2000. Her chest radiographs and CT scans showed partial atelectasis of the right upper lobe, and fiberoptic bronchoscopy revealed bronchial inflammatory changes and whitish mucosal nodular lesions in the walls of the lower trachea, the right main bronchus and the orifice of the right upper lobe bronchus. She was found to have endobronchial tuberculosis. Anti-tuberculosis treatment with isoniazid, rifampicin, streptomycin and pyrazinamide was started. Serum levels of interferon-gamma were markedly elevated on admission. Asthma symptoms improved for a period of one month after the beginning of anti tuberculosis treatment, despite the termination of inhaled corticosteroid. However, as the tuberculosis improved, the frequency and severity of the asthma increased and so corticosteroid inhalation was started again. Four months after administration of the anti-tuberculosis drug, fiberoptic bronchoscopy revealed that the endobronchial lesions had improved without any stenosis or constrictive changes. It was speculated that high doses of inhaled corticosteroid may have the potential to cause endobronchial tuberculosis whilst, ironically, at the same time preventing bronchial stenosis by endobronchial tuberculosis. This is an interesting case in which the asthma symptoms first decreased during the acute phase of endobronchial tuberculosis and then increased again after the tuberculosis improved. PMID- 11729693 TI - [Aggressively metastatic neuroendocrine carcinoma in the middle mediastinum]. AB - A 50-year-old male patient with no symptoms was admitted to our hospital for further evaluation of a right superior mediastinal mass shadow found in chest radiographs during a health examination. Computed tomographic (CT) scans demonstrated a middle mediastinal mass of 4 cm in diameter surrounded by the SVC, the ascending aorta and the trachea at the subaortic level. Routine examination disclosed no metastatic or primary lesion elsewhere. A surgical biopsy resulted in a diagnosis of neuroendocrine carcinoma of unknown primary organ. The patient was given 1 course of chemotherapy with CAV-PE, but little effect was seen. However, when a PE regimen combined with thoracic radiation was used, the tumor shrank to half of its size. After the patient subsequently received high-dose treatment with Etoposide, CT imaging and cytologic analysis revealed multiple metastases to the subcutaneous tissue, both adrenal glands and the liver. Lung lesions also spread aggressively, and the patient died of metastasis. Although neuroendocrine carcinomas can arise from the systemic neuroendocrine cells including those in the mediastinum, their appearance in the middle mediastinum is rarely reported. PMID- 11729694 TI - [A case of pulmonary alveolar proteinosis complicated with pneumoconiosis and myelodysplastic syndrome]. AB - A 72-year-old man who had worked as a coal worker for 28 years and as a tunnel construction worker for 18 years was admitted because of fever, dyspnea, and appetite loss. Pneumoconiosis had been diagnosed when he was 64 years old and myelodysplastic syndrome at 71 years of age. After admission, the patient was treated with antibiotics and anti-fungal drugs but did not respond. Respiratory failure gradually worsened and he died. Autopsy specimens revealed that the cause of death was exacerbation of respiratory failure due to pulmonary proteinosis rather than pulmonary infection. This is a case of pulmonary proteinosis complicated with pneumoconiosis and myelodysplastic syndrome which, to our knowledge, is rare. We also considered the possibility that defective pulmonary macrophage function due to myelodysplastic syndrome and long-term silica inhalation played a part in the development of pulmonary alveolar proteinosis in this case. PMID- 11729695 TI - [Two cases of visceral larva migrans due to Ascaris suum showing a migratory nodular shadow]. AB - The number of cases of visceral larva migrans caused by the pig ascarid, Ascaris suum has recently been increasing. We have encountered two cases of visceral larva migrans due to A. suum with a nodular shadow on the chest radiograph and eosinophilia in the peripheral blood. Patient 1 was a 26-year-old man who had been admitted to our hospital for an elective minor operation. His chest radiology and chest computed tomography revealed a nodule in the left lung field. Peripheral blood eosinophil counts and serum IgE values were elevated. Radiological abnormality disappeared without treatment. Patient 2 was a 57-year old man who had been admitted to our hospital because of a migratory nodule on chest radiography and eosinophilia in the peripheral blood. The diagnosis of visceral larva migrans caused by A. suum was made because the serum of both patients was positive for an antibody against A. suum. Patient 1 and patient 2 were accustomed to eating the raw flesh of wild boar and deer, and of chicken and turkey, respectively. Treatment with albentazole was effective in these patients. PMID- 11729696 TI - Reducing overclaiming in needs assessment studies. An experimental comparison. AB - A randomized experiment investigated the effect of various instructional sets on reducing agency awareness overclaiming, that is, claiming knowledge of fictitious agencies. As predicted, respondents who were warned that the list contained fake agencies exhibited less agency awareness overclaiming than respondents who were not warned. However, providing respondents a memory retrieval strategy had no effect on agency awareness overclaiming. A multivariate model, which included demographic variables, response style variables, and knowledge variables, explained 40% of the variance of agency awareness overclaiming. PMID- 11729697 TI - Characteristics of responding-, nonresponding- and refusing-parents in an adolescent lifestyle choice study. AB - The acquisition of a random sample is one of the many methodological problems that arise when conducting research with adolescent populations. Frequently, due to ethical considerations associated with collecting data from adolescents, active parental consent procedures are required. The current study examined characteristics of parents who consented, refused consent, or did not respond to an active consent request for their children to participate in a large-scale study of adolescent lifestyle behaviors. Results indicated nonresponding-parents were more likely to be employed than consenting-parents. Further, differences were found for a number of attitudinal variables and about the importance of adolescent research. There were significant differences between refusing-parents, and consenting- and nonresponding-parents who were similar in their attitudes toward adolescent research. The findings suggest that nonresponding-parents are characteristically more similar to consenting-parents than to refusing-parents, which supports the use of passive consent procedures as a reasonable alternative to requiring active parental consent in adolescent research. PMID- 11729698 TI - Step to college. Moving from the high school career academy through the 4-year university. AB - This study addresses the question "Do school-to-work programs, as embodied by career academies, facilitate postsecondary education?" The author conceptualizes postsecondary education as a series of steps through the university and examines the high school career academy's influence on entrance into, route through, and outcomes from a 4-year university. Data are drawn from applicant and student records at a comprehensive, urban university for all individuals originating from a single district's high schools. The findings suggest that students from career academies have higher academic achievement upon leaving high school, less need for remediation in English at the university, and a 4-percentage-point increase in graduation from the university than students who are not from academies. These findings suggest that school-to-work programs could facilitate positive outcomes in postsecondary education. However, the continued high rates of remediation and the low rates of graduation, even for students from career academics, suggest that their influence might not be enough to ensure success in postsecondary education. This analysis therefore suggests that further research should identify program components that increase postsecondary education and determine how these components can be institutionalized and built on in subsequent reforms. PMID- 11729699 TI - Assessing the effects of a school-based intervention on unscheduled school transfers during elementary school. AB - Raising Healthy Children is a cluster-randomized study of a school-based intervention aimed at preventing problem behaviors among children recruited into the project in the first or second grade of elementary school. Multilevel analysis was used to compare students in intervention and control schools with respect to whether they transferred out of their original schools. Students in intervention schools were less likely to transfer within the first 5 years of the project. A multilevel discrete-time survival model that included both time varying and contextual variables revealed that the difference in hazard of transfer was greatest in the earlier years of the project. PMID- 11729700 TI - A 5-year-old boy with a chest lesion. PMID- 11729701 TI - Growth hormone therapy in children with growth hormone deficiency. PMID- 11729702 TI - Treatment guideline of enuresis in Taiwan. AB - The treatment guideline of enuresis suggested by the study group of enuresis in Taiwan is reported. Medical consultation and treatment of enuresis are recommended at the age of 5 and 6 years respectively. Evaluation of the enuretic children includes history taking, physical examination and laboratory investigations. A checklist is provided for quick evaluation in busy clinics. Urinalysis, urine specific gravity and office ultrasonography are essential tests. An algorithm of treatment of monosymptomatic enuresis is constructed. The two main options of treatment are behavioral modification with alarm system and pharmacotherapy with desmopressin or imipramine. Imipramine is recommended as one of the treatment options because of its relative safety in Taiwan. However the potential toxicity and mortality of imipramine is highlighted and warned. The responses to treatment are evaluated after a period of treatment for 1 to 3 months. Further evaluation and individualized treatment is suggested for poor responders. PMID- 11729703 TI - Helicobacter pylori infection in recurrent abdominal pain in children--a prospective study. AB - A total of 40 children was recruited to assess the role of Helicobacter pylori infection in children with recurrent abdominal pain syndrome. Among 40 children, seven (17.5%) cases were confirmed to have H. pylori infection. All H. pylori positive patients had active chronic gastritis histologic findings (p < 0.0001); however, the majority of the H. pylori-negative patients had minimal to mild gastritis histologic findings (p = 0.001). Grossly, chronic gastritis picture was present in all children infected with H. pylori and antral nodular gastropathy present in 43%. 71% of H. pylori-positive patients had elevated anti-H. pylori IgG titer; however, 15% in H. pylori-negative patients (p = 0.006). Serum H. pylori assay had a sensitivity of 71% and specificity of 85%; however, sensitivity and specificity of rapid urease test were 86% and 100% respectively. Antral nodularity is a specific, peculiar endoscopic finding of children infected with H. pylori (p = 0.004). Although the present study suggests that H. pylori related chronic gastritis may play an etiological role in a subgroup of children with RAP syndrome, but the routine screening of H. pylori infection in children with RAP is not recommended, since no compelling data support the significant association between H. pylori infection and RAP syndrome. PMID- 11729704 TI - Glucose-insulin infusion for the early treatment of non-oliguric hyperkalemia in extremely-low-birth-weight infants. AB - In order to investigate the status of non-oliguric hyperkalemia and to evaluate glucose-insulin infusion treatment among extremely-low-birth-weight (ELBW) infants, 161 infants weighting less than 1000 gm at birth were enrolled for this study. They were divided into two groups: a hyperkalemic group and a non hyperkalemic group. Hyperkalemia was defined here as a serum potassium level of greater than 6 mEq/L in a non-hemolyzed arterial blood sample. A glucose-insulin infusion was administered to the patients when hyperkalemia was detected in them during the first few days after birth. The infusion was discontinued when the serum potassium levels had been less than 6 mEq/L and stabilized for 6 hours. The incidence of non-oliguric hyperkalemia among ELBW infants in this study was 58% (93/161). The mean gestational age of neonates was 25.7 +/- 1.8 weeks (hyperkalemic) and 26.6 +/- 1.7 weeks (non-hyperkalemic). The mean rate of increases in serum potassium levels was 0.32 +/- 0.29 mEq/L/hr (hyperkalemic) and 0.13 +/- 0.12 mEq/L/hr (non-hyperkalemic). The incidence of severe intraventricular hemorrhage (IVH) was 19% (18/93) (hyperkalemic) and 4.4% (3/68) (non-hyperkalemic). The incidence of cardiac arrhythmia was 12% (11/93) (hyperkalemic) and 0% (non-hyperkalemic) respectively. Neonates with fewer weeks of gestation at birth and faster increases in serum potassium levels were associated with a more prominent tendency toward hyperkalemia. Hyperkalemia markedly increases the risk of severe IVH and arrhythmia for ELBW infants. A higher glucose infusion rate should be maintained to prevent hypoglycemia following insulin treatment. PMID- 11729705 TI - Evaluation of different nebulized bronchodilators on clinical efficacy and hypokalemia in asthmatic children. AB - Acute asthma attack continues to be a major cause for children admitted to the emergency room. Nebulized beta 2-adrenergic agonists are still the first-line drugs for a rapid bronchodilation effect and an easily administered drug during acute asthma attack. The bronchodilator-induced hypokalemia is thought significant in adult group, but is often ignored in children group. In this study, we conducted a randomized study to compare the laboratory and clinical effects between nebulized salbutamol and terbutaline. We found that both salbutamol and terbutaline nebulization induced a significant hypokalemia (p < 0.05). Terbutaline nebulization also significantly improved the peak expiratory flow rate (PEFR) and respiratory rate (RR), but not venous partial pressure of oxygen (PvO2), venous carbon dioxide tension (PvCO2) and O2 saturation in venous blood (SvO2). In contrast, salbutamol improved not only PEFR and RR, but also PvO2, PvCO2, and SvO2 30 minutes of administration. In conclusion, the nebulized salbutamol, although induced a hypokalemic effect which is similar to terbutaline group, has a better effect on improving O2 saturation than nebulized terbutaline with the same dose 30 minutes after administered for children with acute asthma attack. PMID- 11729706 TI - Growth response and final height in growth hormone-deficient patients treated with biosynthetic growth hormone. AB - Twenty-three patients with growth hormone deficiency (GHD) followed until their final heights (FH) were reported by retrospective review. Seven patients had spontaneous puberty (group 1) and sixteen required induction of puberty (group 2). Their heights prior to growth hormone (GH) therapy were -3.7 +/- 1.4 and -2.8 +/- 1.4 SDS in men and women respectively. The mean ages at initiation of GH therapy were 17.2 +/- 3.3 in men and 13.0 +/- 1.7 years in women, with a growth velocity < 4 cm/year and a mean bone age (BA) of 12.6 +/- 0.7 years (men) and 9.8 +/- 1.2 years (women). The dose of GH was 0.27-0.83 IU/kg/week, with a total duration of 2.5 +/- 1.0 years. Their consecutive annual mean growth velocities on GH therapy were: 10.6 cm/year, 8.0 cm/year, 6.1 cm/year, 5.1 cm/year and 4.6 cm/year respectively. They reached a mean final height of 167.1 +/- 5.3 cm (-0.4 +/- 1.0 SDS) in men and 157.9 +/- 3.5 cm (-0.1 +/- 0.7 SDS) in women, which is slightly taller than their target height. Demographic factors related to growth response and final heights were analyzed. We conclude that GH therapy is very effective in linear growth promotion. Their final height was correlated with initial height SDS, target height SDS, predicted adult height SDS according to the bone age at the start of GH therapy and height SDS at the onset of puberty. PMID- 11729707 TI - Pediatric Salmonella enterocolitis in a teaching hospital in Taitung: A four-year analysis. AB - In order to understand the childhood Salmonella enterocolitis in Taitung, we retrospectively collected the patients with the diagnosis of acute enterocolitis who were admitted to the pediatric wards of Taitung branch of Mackay Memorial Hospital from January 1, 1995 to December 31, 1998. Salmonella enterocolitis accounted for 16.8% of the total cases of acute enterocolitis. The mean age of the patients was 14.6 +/- 14.1 months old. Seventy-nine patients were male and 78 patients were female. Eighty-two patients lived in the urban area and 75 patients lived in the suburbs. The mean duration from onset of illness to admission was 2.6 +/- 1.9 days. The mean duration of hospitalization was 6.6 +/- 4.8 days. The peak incidence was in August and September. The most common clinical manifestations were fever (74.5%) and blood stool (46.5%). In 157 patients, Salmonella serogroup B was isolated from stool in 115 patients. The rate of bacteremia was 4.5%. Serogroup D accounted for 28.6% of the bacteremia. Two patients developed meningitis and one patient had osteomyelitis. Of 88 patients examined for rotavirus, 12 had positive results. There was neither bowel perforation nor mortality recorded in our study. PMID- 11729708 TI - Oral bacterial therapy promotes recovery from acute diarrhea in children. AB - Lactobacilli in the intestines play an important role in developing natural defenses against both intestinal bacterial and viral infections. So a prospective clinical study was carried out at Cathay General Hospital to determine the effect of Lactobacillus acidophilus and Bifidobacterium infantis on the course of acute diarrhea in hospitalized children. Altogether 100 children between 6 and 60 months of age were collected and randomly allocated into 2 groups. Study group (n = 50) was given Infloran Berna, which contains 10(9) viable Lactobacillus acidophilus and 10(9) Bifidobacterium infantis, one capsule tid for 4 days and control group (n = 50) received parenteral rehydration only without any medication. Only 20 stool cultures in study group had positive culture results for Lactobacillus or Bifidobacterium. All children were evaluated for the degree of dehydration before rehydration. The clinical course of diarrhea was followed during the treatment period. Features on admission were similar between the study group and control group in age, duration of diarrhea at home, serum sodium & potassium and dehydration degree. The duration of diarrhea was defined as the time until the last appearance of watery stool. There was no difference between the study group and control group in the frequency of diarrhea stools on the day before admission (p > 0.05). However, the frequency of diarrhea for study group improved on the first and second day of hospitalization with statistical difference (p < 0.01). The duration of diarrhea during hospitalization in study group also decreased (3.1 vs. 3.6 days, p < 0.01). Oral bacterial therapy is an effective adjuvant therapy in rotavirus positive and negative children with diarrhea and can safely be administered during an episode of acute diarrhea. PMID- 11729709 TI - Clinical experience in anesthetic management for children with mucopolysaccharidoses: Report of ten cases. AB - The mucopolysaccharidoses (MPS) are hereditary disorders. Children with these disorders have multisystem disease and present significant challenges for the anesthesiologist. The establishment and maintenance of an adequate airway represents the most commonly encountered anesthesia-related problem in these patients. We report ten children with MPS who required surgical intervention. One patient died and the others had a good outcome. In this report, we share our experience and discuss the anesthetic risks and management of MPS patients. PMID- 11729710 TI - Congenital diaphragmatic hernia diagnosed by magnetic resonance imagine in late infancy: Report of one case. AB - We report the magnetic resonance imagine (MRI) findings of an 11-month-old baby with delay onset of congenital diaphragmatic hernia (CDH). The capability of visualizing diaphragmatic discontinuity, connecting bowel segments between abdomen and chest are clearly demonstrated. The various techniques of diagnosing diaphragmatic hernia are described and we emphasize the potential usefulness of MRI in CDH. PMID- 11729711 TI - Intravenous gamma-globulin therapy in myocarditis complicated with complete heart block: Report of one case. AB - Myocarditis complicated with complete heart block is rare in childhood. We report a case of 4-year-old child presented with complete heart block which may have been caused by Mycoplasma pneumoniae. Under emergent temporal pacing, patient experienced cardiogenic shock with pulmonary edema eventually. The cardiopulmonary function was improved with atrial rhythm at the 6th hour later after intravenous infusion with high-dose gamma-globulin (IVIG). The IVIG therapy may have immunomodulatory effects and serve as a potential adjunctive therapy for fulminant myocarditis. PMID- 11729712 TI - [Antimicrobial activities of carbapenems and fourth generation cephems against clinically isolated strains]. AB - We determined the antibacterial activities of 4 carbapenems (imipenem, panipenem, meropenem and biapenem) and 2 forth generation cephems (cefozopran and cefepime) against 850 bacterial strains (18 species) isolated during the period from January 1998 through September 2000. The 4 carbapenems showed excellent antibacterial activities against methicillin-susceptible S. aureus (MSSA), S. pneumoniae, E. faecalis, P. aeruginosa, M. (B). catarrhalis, B. fragilis and Peptostreptococcus spp. as compared with the cephems except the activity of panipenem against P. aeruginosa. Meropenem showed the highest antibacterial activity against 10 species of all Gram-negative strains determined. The antibacterial activities of 2 forth generation cephems against 6 species of Enterobacteriaceae were equal to or higher than those of carbapenems except meropenem. All drugs showed low antibacterial activities against methicillin resistant S. aureus (MRSA). PMID- 11729713 TI - [Pyogenic sacroiliitis in pregnancy]. AB - Pyogenic sacroiliitis is an extremely rare disease in the field of obstetrics and gynecology, and especially for the cases discovered and treated during pregnancy, only five cases can be found in literature. We experienced one case of the disease in which the patient needed urgent hospitalization due to dysbasia caused by fever and pain at the left hip at the 27th week of her pregnancy. The patient was a 31-year-old primipara presenting typical clinical symptoms of pyogenic sacroiliitis along with evidence of severe infection as represented by fever of 39.7 degrees C and CRP of 12.6 mg/dl. She showed a good response to meropenem (MEPM) at 1 g twice a day for 8 days and then at 0.5 g twice a day for 2 days, followed by faropenem (FRPM) at 200 mg three times a day for 12 days, which successfully improved her subjective and objective findings as well as her laboratory test values, resulting in a complete cure. The definitive diagnosis of the disease in the patient was made on the basis of MRI findings, but no pathogen was identified. The patient was found to have marginal placenta previa as a complication, but she had an uneventful trans-vaginal delivery at the 37th week of her pregnancy and left hospital after both she and her baby showed favorable post-delivery progress. The case reported here is the first case of pyogenic sacroiliitis that has ever been discovered and treated during pregnancy in Japan. PMID- 11729714 TI - [Torticollis after unnoticed pharyngeal perforation: suspected retropharyngeal abscess]. AB - Pharyngeal perforations due to foreign bodies are severe when retropharyngeal space infection develops into an abscess. Although the common clinical presentation is sore throat and fever, when the perforation remains occult, torticollis can be a significant symptom. Palatopharyngeal lesions, caused by rigid objects' impactation through the mouth, are common in childhood. Most are not important, but all have the potential hazard of pharyngeal perforation with development of a retropharyngeal abscess. It must be suspected specially when air is shown in the retropharyngeal space on a lateral radiography of the neck, which should always be done. We report a 9-years old boy who had cervical stiffness and torticollis and elevation of the right shoulder, without previous known trauma. Air was shown in the retropharyngeal space on a lateral radiography of the neck in relation to a possible pharyngeal perforation which was later confirmed by a CT scan. We review the literature about diagnosis and treatment of retropharyngeal abscess in childhood. PMID- 11729715 TI - [Laryngeal tuberculosis. Report of 4 cases]. AB - Reporting 4 cases of laryngeal tuberculosis seen between 1992 and 1998 in La Rioja Community. All were men tobacco addicted and alcoholism and since several months presented with dysphonia, odynophagia, dysphagia and lung aspirations. The lesions were localized in epiglottis, vocal fold, back commisure and the subglottic space. Primary complex was pulmonary. All of them were successfully treated with tuberculostsatic drugs, except one who remain with pulmonary aspirations because of a severed epiglottis. PMID- 11729716 TI - [Histologic changes in soft palate in patients with obstructive sleep apnea]. AB - In patients suffering from obstructive sleep apnea syndrome (OSAS) the reiterate injuries against the soft structures of the pharyngeal wall causes macro- and microscopic damages, which differ basically from the alterations verified in subjects without OSAS. We present a comparative histological study of 45 patients with OSAS operated on uvulopalatopharyngoplasties to face up 10 palates coming from normal post-mortem human among which a comparative anatomopathological study of 8 parameters was realized and the differences between the two groups signaled. PMID- 11729717 TI - [Neck schwannomas]. AB - Given the rareness of cervical schwannomas, they are often mistaken for other kinds of lesions. We present 2 clinical cases, make comments on the clinical pathological features involved and point out the essential elements that ensure the differential diagnosis of these tumours. PMID- 11729718 TI - [Descriptive epidemiology of hospital interconsultations carried out at the ORL Service for a year in the San Millan-San Pedro Hospital Complex]. AB - 391 requests of collaboration to our ENT Department during 1996 have been collected. Twelve variables were analysed: number of request, age, sex, date of emission, type (ordinary, with priority, urgent), time elapsed from emission until valuation of the patient, source, reason's request, pathology, complementary tests, treatment and destination (discharge, re-examination during admission or as an outpatient and admission to ENT-Service). RESULTS: 1,58 request were attended per working day, 80 percent were resolved during the first 48 hours, the Internal Medicine Department was the most frequent source, fever without focus or recalcitrant was the most frequent reason from Paediatric Department. We report the epidemiologic features of the most frequent variables. PMID- 11729719 TI - [Impact of chronic seromucous otitis on child linguistic development]. AB - Chronic seromucous otitis (CSO) has been involved in genetic alterations of linguisic infantile development, nontheless studies with regard to our linguistic pattern are not many, as well as those assessing its pattern in the healthy evolvement of a linguistic pragmatism (usage). We fix the level of language skills among 16 children between 5 and 6 year-old, being affected from long lasting CSO. The incidence of alterations was 20 percent in the under of 5, that increases till 33 percent in the sub-group of 6 year-old. These numbers are interesting enough in order to insist in the need of requiringan effective and early treatment of CSO patients and also of scholar implication in its global handling. PMID- 11729720 TI - [Atrophy of the tympanic membrane in contact with hydroxyapatite prosthesis: physiopathologic approach]. AB - The implantation of ossicular replacement prothesis made of dense hydroxyapatite (HA) appears to be a wider method of reconstruction of ossicular chain damaged by suppurative processes or surgery. A review of 19 patients who underwent implantation of HA prosthesis disclosed 4 cases of extrusion and 3 cases of atrophy. When X-ray diffraction analysis of explanted commercially available prosthesis made of dense HA was compared with the results with porous implants and with HS powder employed in the manufacture of both, is suggested that tympanic membrane atrophy could be caused by mechanical and/or chemical factors. PMID- 11729721 TI - [Neurinoma of the descending loop of the hypoglossal nerve. Report of a case]. AB - Neurinomata developed from extracranial portion of hypoglossal nerve are very scarce. We report one case of neurinoma originated in the descending branch of hypoglossal nerve and not having found after careful perusal of the literature of this subject another case we consider the ours as the first portraying this type of pathology and its localization. PMID- 11729722 TI - [Long-term course of laryngeal verrucous carcinoma]. AB - Verrucous carcinoma is an odd kind from squamous or epidermoid carcinoma, amounting between 1 and 3.5% percent of the whole number of laryngeal growths. Its predominant sitting is glottic and are characterized through the pathological anatomy and its good prognosis. We report one case treated in our Department which evolved during 14 years and we point out its good prognosis and other particularities regarding its etiology, pathologic anatomy and diagnosis. PMID- 11729723 TI - [Strategy for post coronary artery bypass grafting in patients with bypass graft stenosis: comparison of percutaneous transluminal coronary angioplasty for the native coronary artery, internal mammary artery and saphenous vein graft]. AB - OBJECTIVES: The strategy for post coronary artery bypass grafting (CABG) was investigated in patients with graft stenosis. METHODS: The study included 123 post-CABG patients with graft stenosis. The patients were divided into three groups according to target vessels; saphenous vein graft (SVG; n = 72), internal mammary artery (IMA; n = 21) and native coronary artery (n = 30). Furthermore, SVG lesions were divided into proximal anastomosis (n = 23), body (n = 40) and distal anastomosis (n = 9). The procedural success rate and late patency rate were compared between the three groups. Furthermore, the relationships between pre percutaneous transluminal coronary angioplasty (PTCA) percentage diameter stenosis, procedural success rate and late patency rate were evaluated. RESULTS: Procedural success rate was similar in the three groups, but late patency rate was higher in the IMA group. Procedural success rate and late patency rate were significantly lower in proximal anastomoses compared to other sites of SVG stenoses, IMA group and native coronary artery group (p < 0.05). Totally occluded native coronary artery lesions had a high procedural success rate compared with occluded IMA and SVG lesions, but the late patency rate was not higher. Procedural success rate showed no significant difference for 75-99% stenotic lesions, but the late patency rate was significantly higher in the IMA group (p < 0.05). Patients in the stenting group had a greater late patency rate compared with the balloon angioplasty group. There was no significant difference in late patency rate between the IMA group and SVG group. CONCLUSIONS: Late patency rate of the IMA is higher than that of the native coronary artery. SVG with proximal anastomosis and severe stenosis shows a significantly lower late patency rate than the native coronary artery. Therefore, PTCA should be considered for the native coronary artery in the absence of chronic total occlusion. PMID- 11729724 TI - [Role of new devices for angioplasty in the unprotected left main coronary artery]. AB - OBJECTIVES: Angioplasty for the unprotected left main trunk remains controversial, but new procedures and devices such as directional coronary atherectomy and stenting have improved the results. We compared the results of angioplasty with the inflexible balloon and new devices. METHODS: The procedures were performed in 239 consecutive lesions in 160 patients between April 1986 and March 2000. This study excluded emergency cases and repeat angioplasty cases, and included 120 initial and elective cases. Initial and long-term results were compared between the balloon group (n = 29) and the new device group (n = 91). RESULTS: Lesion success rate was lower in the balloon group (90% vs 100%, p < 0.05), but there were no significant differences in in-hospital results including cardiac death (0% vs 0%), noncardiac death (0% vs 3.3%), Q-wave myocardial infarction (0% vs 2.2%), non-Q wave myocardial infarction (3.4% vs 6.6%), bypass surgery (0% vs 0%) and repeat angioplasty (6.9% vs 1.1%). Quantitative angiography showed significant improvements in minimal lumen diameter (mean 2.17 vs 3.16 mm, p < 0.001) and percentage diameter stenosis (mean 31% vs 13%, p < 0.001) in the new device group after the procedures. The minimum lumen diameter remained larger at 3 (p < 0.001) and 6 months (p < 0.05) in the new device group. Therefore, angiographic restenosis rate was higher in the balloon group (55% vs 21%, p < 0.005). Five-year survival rate showed no significant difference between the groups (75.0% vs 83.8%). CONCLUSIONS: New devices significantly improved the minimal lumen diameter after angioplasty for unprotected left main trunk disease, and lead to significant improvement of restenosis rate at follow-up. PMID- 11729725 TI - [Clinical significance of the acoustic detection of coronary artery stenosis]. AB - OBJECTIVES: The clinical usefulness was investigated of phonocardiography of coronary artery disease based on stenosis sounds. METHODS: Heart sounds were recorded in 128 patients undergoing coronary angiography and 40 normal young men. Acoustic recordings were made with the patient supine using five sensors affixed to the skin on the left sternal border at the second (L2), third (L3) and fourth (L4) intercostal spaces, and the right sternal border at the second (R2) and third (R3) intercostal spaces. Approximately 400 msec of waveforms immediately following the second heart sound were passed through a band pass filter (400 1,500 Hz) and subjected to frequency analysis using the maximum entropy method. The ratio of heart sound amplitudes within the 400-700 Hz band to those within the 400-1,500 Hz band was designated as the power ratio. RESULTS: The relationship between the degree of stenosis and the stenosis sounds suggested that the power ratio was greater in mild or moderate (50-75%) stenosis. The patients were divided into the stenosis [American Heart Association (AHA) 50 75%], severe stenosis (AHA 90-100%), and normal groups. The power ratio was significantly greater (p < 0.005) in the stenosis group than in the normal and severe stenosis groups for patients with stenosis of the left anterior descending artery or the left main trunk (single-vessel disease). Using a cut-off level of 45%, the sensitivity was 71%, specificity was 65%, and accuracy was 66%. The greatest power ratio difference was recorded at L4, and represented noninvasive detection of mild to moderate stenosis in the left anterior descending artery or left main trunk. No other site showed any statistically significant difference. Detection was impossible in multivessel disease or in cases complicated with valvular disease. CONCLUSIONS: Phonocardiography may allow reliable detection of coronary artery stenosis with mild (AHA 50-75%) stenosis. PMID- 11729726 TI - [Effects of long-term treatment with prostacyclin on plasma adrenomedullin in patients with primary pulmonary hypertension]. AB - OBJECTIVES: This study investigated whether plasma levels of adrenomedullin, a potent vasodilating endogenous neurohumoral mediator, are useful for assessing the severity of primary pulmonary hypertension. METHODS: Seventeen pediatric patients with primary pulmonary hypertension (eight girls, nine boys, mean age 12 +/- 4 years) were enrolled in this study. Thirteen patients in New York Heart Association (NYHA) classes III and IV had been treated with long-term continuous intravenous prostacyclin (PGI2) infusion therapy, and four patients in classes I and II had received beraprost sodium, an oral PGI2 analogue. Blood samples were taken from all patients at the first visit. Plasma levels of atrial and brain natriuretic peptide (ANP, BNP) and endothelin-1, and mature-type adrenomedullin were measured. The relationships were investigated between neurohumoral mediator levels and NYHA class, pulmonary hemodynamics, and exercise capacity assessed by 6-minute walk test. The changes in neurohumoral mediator levels at 1 month, 3 months, and 6 to 12 months were also evaluated in 11 survivors with long-term PGI2 treatment. RESULTS: All neurohumoral mediator levels were positively correlated with severity of NYHA class. Patients in class IV demonstrated significantly elevated neurohumoral mediator levels, except endothelin-1, in comparison with patients in classes I-III. Neurohumoral mediator levels had a significant negative correlation with exercise capacity. Stepwise regression analysis revealed that the BNP to ANP ratio (BNP/ANP) was the most powerful independent factor for total pulmonary resistance (r = 0.85, p = 0.0071) and cardiac index (r = 0.84, p = 0.009). Adrenomedullin was significantly correlated with BNP (r = 0.53, p = 0.03), endothelin-1 (r = 0.66, p = 0.006), and BNP/ANP (r = 0.73, p = 0.0009). ANP and BNP decreased from 196 +/- 213 and 494 +/- 361 pg/ml at baseline to 74 +/- 47 and 153 +/- 133 pg/ml at 1 month, respectively. There was an apparent re-increase in both ANP (187 +/- 194 pg/ml) and BNP (466 +/- 621 pg/ml) at 3 months, regardless of improvement in NYHA class and exercise capacity after long-term PGI2 treatment. In contrast, adrenomedullin decreased from 3.0 +/ 2.2 (baseline) to 1.7 +/- 0.7 fmol/ml at 1 month and 1.6 +/- 0.5 fmol/ml at 3 months. Adrenomedullin was slightly increased at 6-12 months (2.1 +/- 0.9 fmol/ml) without statistical significance. There was a significant relationship between the changes in adrenomedullin at 3 months compared to values at initiation of PGI2 therapy and the changes in mean pulmonary arterial pressure (r = 0.97, p = 0.0041). CONCLUSIONS: Plasma levels of neurohumoral mediators are useful for assessing the severity of primary pulmonary hypertension. In particular, adrenomedullin was valuable for evaluating both cardiac performance and pulmonary hemodynamics after long-term treatment with PGI2 in patients with primary pulmonary hypertension. PMID- 11729727 TI - [Morphological and histological studies of in-stent restenosis in seven types of stents implanted in porcine coronary arteries]. AB - OBJECTIVES: Differences in the mechanism of restenosis after stenting between coil and tubular stents were examined in porcine coronary arteries using histological and immunohistochemical methods. METHODS: Twenty-four pigs underwent balloon-induced injury in the left anterior descending coronary artery. Two weeks later, seven different stents clinically available in Japan (Coil stents: GR I, GR II, Wiktor, Cordis; Tubular stents: gfx, Multilink, Palmaz-Schatz) were implanted in the injured site. Four weeks after the stent implantation, the pigs were sacrificed for histological examination and for morphometrical analysis of the lumen, neointima, media and adventitia. Immunohistochemical studies using anti-proliferating cell nuclear antigen (PCNA), anti-alpha-smooth muscle actin and anti-macrophage antibody were also performed. RESULTS: The coil stents formed eccentric, and the tubular stents formed concentric neointimal proliferation. Although there was no difference in the area of neointima between the stents, the area of lumen in the tubular stents was bigger than that in the coil stents (p < 0.01), because the vascular area was bigger in the tubular stents (p < 0.05). Immunohistochemical examination found many PCNA-positive cells in the proliferated neointima, especially in the area around the stent strut. Many of these cells around the stent strut were positively stained by anti-macrophage antibody. Other cells positively stained for PCNA were confirmed as smooth muscle cells. CONCLUSIONS: Tubular stents maintained a wider lumen than coil stents, because negative remodeling after stenting was less in the tubular stents. Implantation of stents with less negative remodeling is very important to prevent restenosis after stenting. PMID- 11729728 TI - [Collateral blood flow showing dissection-like filling defect on coronary arteriography: a case report]. AB - A 51-year-old man presented under a diagnosis of angina pectoris manifesting as exertional chest pain. First coronary angiography showed severe stenosis with ulceration and spontaneous dissection at the proximal right coronary artery and linear dissection-like filling defects extending to the distal right coronary artery. After about 3 months, repeat coronary angiography showed the previously observed stenosis with unclear dissection, and better developed collaterals from the left coronary artery to the right coronary artery showing the linear dissection-like filling defects. The bilateral coronary angiography did not clearly show filling defects. This phenomenon suggested that the collateral flows were related to filling of the defects. Intravascular ultrasonic imaging demonstrated severe atherosclerotic lesions at the proximal right coronary artery, but no dissection in the distal right coronary artery. Percutaneous transluminal coronary angioplasty for the stenosis was performed successfully with a stent. Coronary angiography after the angioplasty showed no collaterals, and the right ventricular branch appeared, suggesting that the linear dissection like filling defects extending to the distal right coronary artery were due to the collateral flows. Filling defects extending distal to a severe stenosis must be distinguished carefully from coronary dissection. PMID- 11729729 TI - [A 66-year-old woman with post-operative pulmonary valve stenosis showing spontaneous echo contrast in the right atrium]. PMID- 11729730 TI - [Intraocular lenses]. PMID- 11729731 TI - [The life work of Sir Nicholas Harold Lloyd Ridley]. AB - On may 25th 2001 Sir Harold Ridley died at age 94. With him Ophthalmology lost one of the greatest and most influential Ophthalmologist of the post-war time. He is the founder of modern cataract surgery implanting the first intraocular lens after extracapsular cataract extraction 50 years ago. He pioneered in what became one of the most important success stories in Ophthalmology. His vision and concepts managed to succeed against strong opposition worldwide. Modern cataract surgery and modern refractive IOL surgery would not been possible without his contribution. Sir Harold Ridley reached a high age and was therefore able to see his visions become reality and finally receive all honours and awards he deserved over the past 15 years, including the knighthood by Queen Elisabeth II in the year 2000. The following article gives a summary of his biography and contributions to Ophthalmology. PMID- 11729733 TI - [Complication profiles of posterior chamber intraocular lenses IOL). An analysis of 586 foldable and 2077 rigid explanted intraocular lenses]. AB - BACKGROUND: Foldable intraocular lenses (IOL) have become increasingly preferred choice for IOL implantation after cataract removal. However, both foldable as well as rigid IOLs are not yet complication-free and may need explanation. MATERIAL AND METHODS: A total of 2663 explanted posterior chamber IOLs (PCIOLs) were accessioned at the Center for Research on Ocular Therapeutics and Biodevices between January 1988 and September 2000. The lenses were examined grossly using a Leitz/Wild M-8-Zoom stereomicroscope. The clinical reasons for explanation were documented for foldable as well as for rigid lenses. RESULTS: Of a total of 2663 explanted PCIOLs, 586 were foldable lenses and 2077 were rigid PMMA lenses The most frequent reason for explantation of all 2663 IOLs studied was decentration/dislocation. Optic and haptic damage and posterior capsule rupture were significantly more often a reason for explantation in several foldable designs compared to rigid PCIOLs. Whereas rigid designs lead more often to corneal decompensation. The percentage of explanted IOLs because of inflammatory reactions decreased significantly from 1994 to 2000. CONCLUSIONS: The complication profiles of rigid and foldable IOLs revealed some apparent differences due to the nature of the IOL biomaterials and designs. IOL optic/haptic damage was a common complication for foldable IOLs, whereas it was only discovered in single digits for rigid PMMA IOLs. The possible explanation for this is, that the soft and flexible biomaterials, from which all the foldable IOLs were manufactured, are easier to be damaged during folding, unfolding and insertion procedure. In our opinion this relative disadvantage of foldable IOLs is by far outweighed by the numerous advantages of the small incision cataract surgery. The decreasing inflammatory reactions can be explained by advances in surgical techniques, especially the secure in-the-bag fixation of IOLs. PMID- 11729732 TI - [History of the development of intraocular lenses]. AB - Even though cataract surgery has been practiced for over 2000 years, modern cataract surgery started just some 50 years ago. with the first IOL implantation by Sir Harold Ridley. The development of intraocular lenses was accompanied by great successes and disasters. With the fast development of cataract surgical techniques over the past 15 years (ECCE, Phacoemulsification, Capsulorhexis) a successful marriage between IOL-developments and surgery was established. Indication profiles for cataract surgery and IOL implantation extended to more and more patient groups. At this time classical cataract surgery is further developing into refractive intraocular lens surgery to correct higher ametropia in clear lens or phakic eyes. This development was only possible because of the improvements of surgical techniques and implants in classical cataract surgery. PMID- 11729734 TI - [Morphological autopsy findings in human eyes implanted with Ridley intraocular lenses]. AB - BACKGROUND: Sir Harold Ridley's first cataract extraction with implantation of an intraocular lens (IOL) marked the beginning of a major change in the practice of ophthalmology. MATERIAL AND METHODS: Two human autopsy globes implanted with original Ridley IOLs were grossly examined from behind with the Miyake-Apple photographic technique. The anterior segments were evaluated histologically. One Ridley IOL was examined by scanning electron microscopy. RESULTS: Both IOLs maintained a clear visual axis and had been placed in the capsular sac. The right lens was decentered inferiorly, the left IOL was well centered. The posterior capsule showed only minimal capsular opacification. Scanning electron microscopy (SEM) of the explanted IOL showed a smooth, well polished surface. CONCLUSIONS: In this patient Ridley IOLs maintained an ideal visual rehabilitation of 20/20 in both eyes, 22 and 18 years postoperatively. One surprising finding was the presence of a square, truncated optic edge geometry, very similar to that seen in some modern foldable IOLs. In our opinion the absence of posterior capsule opacification of the Ridley IOL may have partially been a result of this nearly sharp optic edge, that we confirmed by SEM. Even 22 years after implantation the Ridley IOL showed a high finish quality. Many of Sir Harold Ridley's early concepts like the IOL placement in the posterior chamber and endocapsular implantation are now the standard of modern cataract surgery. PMID- 11729736 TI - [Glare and halos after "phakic IOL". Surgery for the correction of high myopia]. AB - BACKGROUND: Phakic IOLs are currently under clinical investigation for the correction of high myopia with acceptable postoperative refractive results. Although daytime vision is usually very good some patients complain of visual disturbances, described as glare and halos, during the night time, leading to difficulties in driving vehicles. PATIENTS AND METHODS: The quality of vision after phakic IOL surgery was investigated pre- and postoperatively using various tests for objective measurements of glare and halo in 40 myopic eyes (-9.0 to 20.0 D). Based on the implanted phakic IOL, eyes were divided into 4 groups of 10 eyes each where an Artisan 5.0 was implanted, an Artisan 6.0, a NuVita, and an ICL. RESULTS: All patients were happy with the postoperative refractive outcome and during daytime no visual discomfort was reported. In contrast, most patients reported visual disturbances during dim light conditions and at night time, described as glare and halos. The objective measurements showed in most eyes increased glare and halos which was most prominent in eyes with an ICL, followed by the NuVita IOL. In contrast, patients with an Artisan 6.0 reported significantly less glare and halo problems. CONCLUSIONS: All currently available phakic IOLs, in particular the ICL and the NuVita lead to a decreased visual performance during night time. Therefore, this phenomenon must be explained to the patient prior to surgery. An increase in the size of the optic should lead to an improvement with reduction of postoperative glare and halos. PMID- 11729735 TI - [LASIK with a superior hinge and scanning spot excimer laser ablation for correction of myopia and myopic astigmatism. Results of a prospective study on 100 eyes with a 1-year follow-up]. AB - PURPOSE: Recently laser-in-situ-keratomileusis (LASIK) has been increasingly used to correct refractive errors. The purpose of this investigation was to evaluate the safety, efficacy, predictability, stability and complications using the scanning spot excimer LASIK technology. PATIENTS AND METHODS: The results of 100 consecutive LASIK operations carried out between 2/1998 and 2/1999 on 60 patients (mean age 37 years, range 20-55 years) have been evaluated. The Hansatome microkeratome with a superior hinge (flap diameter: 9.5 mm) and a scanning spot excimer laser (Technolas C-Lasik 217) were used in all cases. The mean spherical equivalent of the subjective manifest refraction was -6.96 +/- 2.87 diopters (D), the mean sphere was -6.47 +/- 2.71 D and the mean astigmatism was -0.98 +/- 0.94 D. In 6% of the eyes preoperative visual acuity was not better than 20/40. Examinations were performed preoperatively, after 1 and 7 days, after 1,4 and 12 months. Safety, efficacy, predictability, stability and complications were calculated using the datagraph software (version 1.11). RESULTS: All 100 eyes were examined after 1 day and 1 week, 96 after 1 month, 95 after 4 months and 92 after 1 year. Following 1 year none of the eyes lost 2 or more lines of best corrected visual acuity, 99% were within +/- 1 line and 1% gained 2 lines (safety index 1.03). In 92% of all eyes an uncorrected visual acuity of > or = 0.5 was reached, in 77% > or = 0.8 and in 51% > or = 1.0 (efficacy index 0.89). For 60 eyes (65.21%) a refractive correction of +/- 0.5 D was necessary, for 82 eyes (89.13%) +/- 1.0 D and for 91 eyes (98.91%) +/- 2.0 D. The mean spherical equivalent after 1 year was -0.15 +/- 1.31 D. Between 1 and 12 months a mean regression of -0.14 D occurred. On the request of the patients, 5 eyes were retreated during the study period for under- or over-correction. Complications due to the microkeratome did not occur. Corneal infections were not observed, a diffuse lamellar keratitis (DLK) was seen in 12 cases, but all cases healed with no loss of visual acuity. In 7 eyes a slight decentration of the ablation was observed using corneal topography, which in one case caused an increase in glare sensitivity. CONCLUSIONS: LASIK with superior hinge and scanning spot excimer photoablation is suitable for the correction of myopia (up to a maximum of -12 D) and for myopic astigmatism (up to a maximum of -5 D). The refractive results showed a high stability during the 12-month study period but there is still room for improvement of the predictability. PMID- 11729737 TI - [Preoperative scanning laser ophthalmoscopy: findings in idiopathic macular foramen]. AB - BACKGROUND: The aim of our study was a detailed morphological and functional assessment of macular foramen stages II-IV by scanning laser ophthalmoscopy (SLO) and fundus perimetry including a correlation with clinical findings. METHODS AND MATERIALS: Included in this prospective study were 78 patients where SLO imaging and fundus perimetry were performed with the SLO-105 (Rodenstock) prior to pars plana vitrectomy for macular foramen on one eye. Both the size and shape the of macular foramen were assessed. Deep (0 dB) and relative (12 dB) scotomata were tested using the Goldmann II stimulus in the 20 degrees field of the SLO. Imaging was performed with the infrared laser and confocal aperture C2. RESULTS: The size of macular foramen ranged from 140 microns to 800 microns (median: 420 microns) and 20 (26%) had irregular, polycyclic borders. In 21 patients (27%) a bright sector could be seen just outside the hole. Deep scotomata were notably smaller than the visible macular hole in 4 patients (6%) and 18 deep scotomata (25%) extended beyond the hole. There was a weak inverse correlation between the size of deep, relative scotomata and the diameter of the macular holes with the visual acuity (r = -0.46; r = -0.39 or r = -0.53), but no correlation with the duration of symptoms could be demonstrated. CONCLUSION: Polycyclic borders were seen as a new feature of some idiopathic macular foramen. This may be due to asymmetric traction or adhesions on the macular area. Also, deep scotomata either smaller or larger than the visible macular hole were found. PMID- 11729738 TI - [Population-based study of diabetic retinopathy in Wolfsburg]. AB - INTRODUCTION: Since November 1997 the complete documentation of an ophthalmological examination of diabetics has been annually subsidized by the Volkswagen Corporation Health Maintenance Organization (VW-HMO). METHODS: The results of an annual ophthalmological examination were recorded in a standardised history sheet developed by the Initiative Group for Early Detection of Diabetic Eye Diseases. These data included visual acuity, intraocular pressure, lens status and a description of fundus abnormalities. RESULTS: Within 26 months ophthalmological examinations of 2,801 patients were completed which represented 4.5% of all VW-HMO insured patients. On average, patients suffered from diabetes for 9.6 years (SD +/- 8.3), artificial intraocular lenses were present in 357 eyes (6.4%) and 1,216 eyes (12.0%) were diagnosed with cataract or posterior capsule opacification impairing visual acuity. Out of 263 patients younger than 40 years old, 18.8% had a mild or moderate and 3.3% a severe non-proliferative diabetic retinopathy (NPDR). A proliferative diabetic retinopathy (PDR) was found in 2.2% of the younger patients. Of 2,228 patients aged 40 years and older, 11.9% had a mild or moderate and 2.6% a severe NPDR. In 0.9% of this group PDR was diagnosed. CONCLUSIONS: An annual ophthalmological screening based on a survey sheet of the Initiative Group was successfully introduced. For the first time a population-based evaluation on the prevalence of diabetic retinopathy was carried out for inhabitants of a German city. The prevalence of PDR was found to be lower than previously published in comparable studied. PMID- 11729739 TI - [Pathogenesis of central serous chorioretinopathy: angiographic and electrophysiological studies]. AB - BACKGROUND: By the combination of fluorescein and indocyanine green angiography and multifocal electroretinography (mERG) we examined the correlation of angiographic and electrophysiological findings in acute central serous choroiditis (CSC) to find out if this leads to further information about the course of the disease. MATERIAL AND METHODS: Patients with unilateral, acute CSC (n = 49) were examined by the use of fluorescein and indocyanine green angiography and mERG. The examinations were repeated after 6 and 12 weeks. RESULTS: Of the patients 47 (96%) showed hypofluorescent areas in the early pictures of the ICG angiography which changed to hyperfluorescence in the late phase pictures in 39 patients (80%). Of the asymptomatic paired eyes, 31 (63%) showed abnormalities in the ICG pictures. The amplitudes in mERG were reduced in the study eyes as well as in the paired eyes and only recovered slowly. Angiographic healing was seen in 44 (90%) patients during the follow-up time, while the reduction of mERG amplitudes recovered more slowly and did not reach normal levels. CSC is a bilateral disease of the central choroid with acute decompensation of the retinal pigment epithelium in one eye. This leads angiographically to a local hypofluorescence with subsequent hyperfluorescence caused by barrier breakdown. The functional recovery follows the morphological healing only slowly. The mERG seems to be an adequate measurement method. PMID- 11729740 TI - [Multifocal ERG in central areolar choroidal dystrophy]. AB - BACKGROUND: In comparison to full-field ERG a multifocal ERG (mf-ERG) provides functional mapping of the central retina and detection of more localised defects. Central areolar choroidal dystrophy is an autosomal dominant inherited disorder with central atrophy of the choriocapillaries, the retinal pigment epithelium (RPE) and the photoreceptors. Full-field ERG often shows only slight to moderate reductions. The purpose of our study was to investigate the electroretinographic activity of the posterior pole with the mf-ERG in comparison to the photopic full field ERG in these patients. PATIENTS AND METHODS: We performed a mf-ERG and a photopic full-field ERG in 4 patients with central areolar choroidal dystrophy and in 20 age-matched volunteers who constituted the normal control group. RESULTS: In all patients reductions in b-wave amplitudes of the first order kernel of the mf-ERG in both eyes were evident. Full-field ERG showed decreased b wave amplitudes in only three eyes. Subnormal amplitudes were found in two eyes and normal amplitudes in three eyes. Additionally ophthalmoscopic lesions were found to a lesser extent than by the mf-ERG. DISCUSSION: The mf-ERG is a valuable tool in detecting central areolar choroidal dystrophy and might be useful in early detection and follow-up, when ophthalmoscopy and full-field ERG are still normal. Moreover it could contribute to differential diagnosis of other retinal diseases of the posterior pole. PMID- 11729741 TI - [Eyelid injuries: epidemiological aspects]. AB - PURPOSE: To present epidemiological data on eyelid trauma which appear rarely in the literature. PATIENTS AND METHODS: Medical records of 180 patients given surgical treatment for eyelid injuries at the ophthalmology department of LMU Munich between 1997 and 1999 were evaluated. Clinical findings of eyelids and eyeballs, surgical procedures and complications during wound healing were considered. RESULTS: Out of 180 patients, 140 suffered blunt trauma of which 66 were due to manual work, 38 to sudden falls, 23 to violence, 7 to motor vehicle accidents and 6 to sports. From 26 cutting injuries caused by glass, 12 were caused by broken spectacles, 8 were caused by sudden falls, 4 by violence and only 2 by car accidents. Clinically 85 patients showed injuries of the upper eyelid, 55 of the lower and 40 of both lids. The lid margin was affected in 43 patients and the lacrimal drainage system in 28. Wound contamination occurred in 34 patients, 19 had foreign bodies and 13 loss of tissue. Accompanying injuries of the face was noticed in 16 patients and of the eyeball in 79 patients of which 31 were superficial, 36 were eyeball contusions of varying severity, 12 were perforations or ruptures and 4 patients additionally had orbital bone fractures. The average time from injury to surgery was 3.3 h. A total of 100 patients needed simple skin suturing, 26 sutures to the skin and subcutaneous tissue including the orbicularis and levator muscle, 43 needed readaption of the lid margin and 24 repair of the canalicular system. Post-operatively only one patient developed a necrosis and one an infection. CONCLUSIONS: In eyelid trauma, the lid margin is affected in 24% and the lacrimal drainage system in 16% of all injuries. Our data also showed a high association (44%) of eyelid injuries with trauma to the eyeball. This emphasises the necessity for eyelid injuries to be treated primarily by an ophthalmic surgeon, although difficult primary reconstruction and early complications in particular, are relatively rare. PMID- 11729742 TI - [Electronic versus paper-based patient records: a cost-benefit analysis]. AB - BACKGROUND: The aim of this study is to compare the costs and benefits of electronic, paperless patient records with the conventional paper-based charts. METHODS: Costs and benefits of planned electronic patient records are calculated for a University eye hospital with 140 beds. Benefit is determined by direct costs saved by electronic records. RESULTS: In the example shown, the additional benefits of electronic patient records, as far as they can be quantified total 192,000 DM per year. The costs of the necessary investments are 234,000 DM per year when using a linear depreciation over 4 years. In total, there are additional annual costs for electronic patient records of 42,000 DM. Different scenarios were analyzed. By increasing the time of depreciation to 6 years, the cost deficit reduces to only approximately 9,000 DM. Increased wages reduce the deficit further while the deficit increases with a loss of functions of the electronic patient record. However, several benefits of electronic records regarding research, teaching, quality control and better data access cannot be easily quantified and would greatly increase the benefit to cost ratio. CONCLUSION: Only part of the advantages of electronic patient records can easily be quantified in terms of directly saved costs. The small cost deficit calculated in this example is overcompensated by several benefits, which can only be enumerated qualitatively due to problems in quantification. PMID- 11729743 TI - [Electrical stimulation of the visual system. From empirical approach to visual prostheses]. AB - The investigation of the influence of electrical stimulation on the visual system can be sub-divided into empirical, diagnostic and therapeutic phases of development. The first electrical stimulations of the visual system in animals dating back to the eighteenth century represent empirical approaches well before knowledge on the principles of bioelectrical nerve activity was available. Despite this lack of understanding, attempts were made at that time to treat blind people by electrical stimulation. More than 100 years later, with the advancement of technology and knowledge on nerve function, attempts were made to establish various methods of electrical stimulation for the diagnosis of different diseases of the visual system. For more than 30 years attempts to develop electrical devices have been made to by-pass defects of the visual system and to restore basic orientation. Presently nine different groups world-wide are focusing on retinal, optic or cortical implants. A short review on the historical use of electrical stimulation and new therapeutical attempts are given in this paper. PMID- 11729744 TI - [Iris metastases in breast carcinoma]. AB - BACKGROUND: Metastatic cancer of the iris is rare. We report a case of breast carcinoma which metastasized to the iris and a review of the literature. METHOD: A 60-year-old woman with a history of breast carcinoma presented with lesions of the iris 5 years after diagnosis and tumor excision. In addition to slit-lamp examination including gonioscopy, ultrasound biomicroscopy, positron-emission tomography and fluorescein angiography of the iris were used for diagnosis. Furthermore, the literature was searched using the medline database. RESULTS: The iris metastasis exhibited multiple whitish to pink nodules on the inferior half of the iris and infiltration of the chamber angle. The main tumor mass was prominent and highly vascularized. Best corrected visual acuity was 20/40 and the intraocular pressure was 7 mmHg. Ultrasound biomicroscopy showed lobular masses characterized by mid to low reflectivity and there was no distinct border to the surrounding tissue. Fluorescein angiography of the iris showed tumor vessels with dye leakage. The positron-emission tomography displayed metastatic lesions to the liver, lung, bones and lymph nodules in addition to the lesion of the iris. The review of the literature revealed that breast carcinoma leads to choroidal metastases in 5% of cases and iris metastases are even rarer. In twothirds of these cases the lesions form unilateral whitish to red clumps at the horizontal meridian and in the inferior quadrants of the iris. The preferred treatment is chemotherapy and/or radiation therapy. CONCLUSIONS: Breast carcinoma is rarely associated with iris metastasis. Positron-emission tomography is a sensitive diagnostic tool to identify metastatic lesions and is a useful method for planning therapeutic approaches. Radiation therapy, argon laser treatment and block excision are therapeutic options with an isolated metastasis of the iris. PMID- 11729745 TI - [LEOPARD syndrome with iris-retina-choroid coloboma. Discordant findings in monozygotic twins (MIM # 151 100)]. AB - INTRODUCTION: The LEOPARD syndrome is an autosomal dominant inherited disease with severe lentiginosis associated with various abnormalities such as electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of the genitalia, retardation of growth and deafness. Ocular manifestations such as coloboma of the iris, the retina and the choroid have not been reported so far. PATIENTS: We report the cases of two 10-year-old identical twins and their mother, showing typical manifestations consistent with the LEOPARD syndrome. Additionally, colobomas of the iris, the retina and the choroid were detected. RESULTS: In addition to the findings typical for the LEOPARD syndrome, we observed unusual ocular abnormalities in all three patients. It represents a discordant phenotype in monozygotic twins. CONCLUSION: LEOPARD syndrome is a disease with multiple alterations and abnormalities. Although ocular malformations seem to be rare, an ophthalmological examination is recommended in order to initiate early visual rehabilitation. PMID- 11729746 TI - [Treatment of retinal vein occlusion. Vitrectomy with arteriovenous decompression and dissection of the internal limiting membrane]. AB - BACKGROUND: Our therapeutical armamentarium for functional improvement after branch retinal vein occlusion (BRVO) is still limited and the efficacy is uncertain. Recently, surgical decompression of the vein at the arteriovenous crossing (AV decompression) has again been proposed as a suitable form of treatment. MATERIAL AND METHODS: A total of 40 patients with BRVO were treated in our department from August 1999 to February 2001 with AV decompression and 22 patients with comparable BRVO who refused this surgical intervention served as control. The surgical procedure consisted of a standard pars plana vitrectomy and separation of the overlying artery from the vein using microscissors. In 13 cases dissection of the internal limiting membrane was additionally performed. Assessment of visual acuity, fluorescein angiography and multifocal ERG in 7 patients was performed preoperatively and 6 weeks after surgery. RESULTS: The surgical procedure and postoperative course were uneventful. In most of the treated eyes, visual acuity improved and fluorescein angiography revealed capillary reperfusion. Functional results in patients with AV-decompression were highly significantly better than in the control group. CONCLUSION: Our results with surgical decompression of branch retinal vein occlusions demonstrate the therapeutical effect of surgical AV-decompression in BRVO. Further experience is necessary before some open questions can be answered. PMID- 11729747 TI - [Recurrent spontaneous macular hemorrhage]. PMID- 11729748 TI - [Multifocal electroretinography (mfERG)]. PMID- 11729749 TI - Cooperation of liver cells in health and disease. AB - The liver lobule is formed by parenchymal cells, i.e., hepatocytes and nonparenchymal cells. In contrast to hepatocytes that occupy almost 80% of the total liver volume and perform the majority of numerous liver functions, nonparenchymal liver cells, which contribute only 6.5% to the liver volume, but 40% to the total number of liver cells, are localized in the sinusoidal compartment of the tissue. The walls of hepatic sinusoid are lined by three different cell types: sinusoidal endothelial cells (SEC), Kupffer cells (KC), and hepatic stellate cells (HSC, formerly known as fat-storing cells, Ito cells, lipocytes, perisinusoidal cells, or vitamin A-rich cells). Additionally, intrahepatic lymphocytes (IHL), including pit cells, i.e., liver-specific natural killer cells, are often present in the sinusoidal lumen. It has been increasingly recognized that both under normal and pathological conditions, many hepatocyte functions are regulated by substances released from neighboring nonparenchymal cells. Liver sinusoidal endothelial cells constitute the lining or wall of the hepatic sinusoid. They perform important filtration function due to the presence of small fenestrations that allow free diffusion of many substances, but not of particles of the size of chylomicrons, between the blood and the hepatocyte surface. SEC show huge endocytic capacity for many ligands including glycoproteins, components of the extracellular matrix (ECM; such as hyaluronate, collagen fragments, fibronectin, or chondroitin sulphate proteoglycan), immune complexes, transferrin and ceruloplasmin. SEC may function as antigen-presenting cells (APC) in the context of both MHC-I and MHC-II restriction with the resulting development of antigen-specific T-cell tolerance. They are also active in the secretion of cytokines, eicosanoids (i.e., prostanoids and leukotrienes), endothelin-1, nitric oxide, and some ECM components. Kupffer cells are intrasinusoidally located tissue macrophages with a pronounced endocytic and phagocytic capacity. They are in constant contact with gut-derived particulate materials and soluble bacterial products so that a subthreshold level of their activation in the normal liver may be anticipated. Hepatic macrophages secrete potent mediators of the inflammatory response (reactive oxygen species, eicosanoids, nitric oxide, carbon monoxide, TNF-alpha, and other cytokines), and thus control the early phase of liver inflammation, playing an important part in innate immune defense. High exposure of Kupffer cells to bacterial products, especially endotoxin (lipopolysaccharide, LPS), can lead to the intensive production of inflammatory mediators, and ultimately to liver injury. Besides typical macrophage activities, Kupffer cells play an important role in the clearance of senescent and damaged erythrocytes. Liver macrophages modulate immune responses via antigen presentation, suppression of T-cell activation by antigen-presenting sinusoidal endothelial cells via paracrine actions of IL-10, prostanoids, and TNF-alpha, and participation in the development of oral tolerance to bacterial superantigens. Moreover, during liver injury and inflammation, Kupffer cells secrete enzymes and cytokines that may damage hepatocytes, and are active in the remodeling of extracellular matrix. Hepatic stellate cells are present in the perisinusoidal space. They are characterized by abundance of intracytoplasmic fat droplets and the presence of well-branched cytoplasmic processes, which embrace endothelial cells and provide focally a double lining for sinusoid. In the normal liver HSC store vitamin A, control turnover of extracellular matrix, and regulate the contractility of sinusoids. Acute damage to hepatocytes activates transformation of quiescent stellate cells into myofibroblast-like cells that play a key role in the development of inflammatory fibrotic response. Pit cells represent a liver-associated population of large granular lymphocytes, i.e., natural killer (NK) cells. They spontaneously kill a variety of tumor cells in an MHC-unrestricted way, and this antitumor activity may be enhanced by the secretion of interferon-gamma. Besides pit cells, the adult liver contains other subpopulations of lymphocytes such as gamma delta T cells, and both "conventional" and "unconventional" alpha beta T cells, the latter containing liver-specific NK T cells. The development of methods for the isolation and culture of main liver cell types allowed to demonstrate that both nonparenchymal and parenchymal cells secrete tens of mediators that exert multiple paracrine and autocrine actions. Co-culture experiments and analyses of the effects of conditioned media on cultures of another liver cell type have enabled the identification of many substances released from non-parenchymal liver cells that evidently regulate some important functions of neighboring hepatocytes and non-hepatocytes. To the key mediators involved in the intercellular communication in the liver belong prostanoids, nitric oxide, endothelin-1, TNF-alpha, interleukins, and chemokines, many growth factors (TGF-beta, PDGF, IGF-I, HGF), and reactive oxygen species (ROS). Paradoxically, the cooperation of liver cells is better understood under some pathological conditions (i.e., in experimental models of liver injury) than in normal liver due to the possibility of comparing cellular phenotype under in vivo and in vitro conditions with the functions of the injured organ. The regulation of vitamin A metabolism provides an example of the physiological role for cellular cross-talk in the normal liver. The majority (up to 80%) of the total body vitamin A is stored in the liver as long-chain fatty acid esters of retinal, serving as the main source of retinoids that are utilized by all tissues throughout the body. Hepatocytes are directly involved in the uptake from blood of chylomicron remnants, and the synthesis of retinol-binding protein that transfers retinol to other tissues. However, more than 80% of the liver retinoids are stored in lipid droplets of hepatic stellate cells. HSC are capable of both uptake and release of retinol depending on the body's retinol status. The activity of some major enzymes of vitamin A metabolism have been found to be many times higher per protein basis in stellate cells than in hepatocytes. Despite progress in the understanding of the roles played by these two cell types in hepatic retinoid metabolism, the way in which retinoids move between the parenchymal cells, stellate cells, and blood plasma has not been fully elucidated. Sinusoidal blood flow is, to a great extent, regulated by hepatic stellate cells that can contract due to the presence of smooth muscle alpha actin. The main vasoactive substances that affect constriction or relaxation of HSC derive both from distant sources and from neighboring hepatocytes (carbon monoxide, leukotrienes), endothelial cells (endothelin, nitric oxide, prostaglandins), Kupffer cells (prostaglandins, NO), and stellate cells themselves (endothelin, NO). The cellular cross-talk reflected by the fine-tuned modulation of sinusoidal contraction becomes disturbed under pathological conditions, such as endotoxemia or liver fibrosis, through the excess synthesis of vasoregulatory compounds and the involvement of additional mediators acting in a paracrine way. The liver is an important source of some growth factors and growth factor-binding proteins. Although hepatocytes synthesize the bulk of insulin-like growth factor I (IGF-I), also other types of nonparenchymal liver cells may produce this peptide. Cell-specific expression of distinct IGF-binding proteins observed in the rat and human liver provides the potential for specific regulation of hepatic IGF-I synthesis not only by growth hormone, insulin, and IGF-I, but also by cytokines released from activated Kupffer (IL-1, TNF-alpha, TGF-beta) or stellate cells (TGF-alpha, TGF-beta). Hepatic stellate cells may affect turnover of hepatocytes through the synthesis of potent positive as well as negative signals such as, respectively, hepatocyte-growth-factor or TGF-beta. Although hepatocytes seem not to produce TGF-beta, a pleiotropic cytokine synthesized and secreted in the latent form by Kupffer and stellate cells, they may contribute to its actions in the liver by the intracellular activation of latent TGF-beta, and secretion of the biologically active isoform. Many mediators that reach the liver during inflammatory processes, such as endotoxins, immune complexes, anaphylatoxins, and PAF, increase glucose output in the perfused liver, but fail to do so in isolated hepatocytes, acting indirectly via prostaglandins released from Kupffer cells. In the liver, prostaglandins synthesized from arachidonic acid mainly in Kupffer cells in a response to various inflammatory stimuli, modulate hepatic glucose metabolism by increasing glycogenolysis in adjacent hepatocytes. The release of glucose from glycogen supports the increased demand for energetic fuel by the inflammatory cells such as leukocytes, and additionally enables enhanced glucose turnover in sinusoidal endothelial cells and Kupffer cells which is necessary for effective defense of these cells against invading microorganisms and oxidative stress in the liver. Leukotrienes, another oxidation product of arachidonic acid, have vasoconstrictive, cholestatic, and metabolic effects in the liver. A transcellular synthesis of cysteinyl leukotrienes (LTC4, LTD4, and LTE4) functions in the liver: LTA4, an important intermediate, is synthesized in Kupffer cells, taken up by hepatocytes, converted into the potent LTC4, and then released into extracellular space, acting in a paracrine way on Kupffer and sinusoidal endothelial cells. Thus, hepatocytes are target cells for the action of eicosanoids and the site of their transformation and degradation, but can not directly oxidate arachidonic acid to eicosanoids. (ABSTRACT TRUNCATED) PMID- 11729750 TI - Biochemical engineering of the production of plant-specific secondary metabolites by cell suspension cultures. AB - Plant cell culture has recently received much attention as a useful technology for the production of valuable plant-derived secondary metabolites such as paclitaxel and ginseng saponin. The numerous problems that yet bewilder the optimization and scale-up of this process have not been over emphasized. In spite of the great progress recorded in recent years towards the selection, design and optimization of bioreactor hardware, manipulation of environmental factors such as medium components, light irradiation, shear stress and O2 supply needs detailed investigations for each case. Recent advances in plant cell processes, including high-density suspension cultivation, continuous culture, process monitoring, modeling and control and scale-up, are also reviewed in this chapter. Further developments in bioreactor cultivation processes and in metabolic engineering of plant cells for metabolite production are expected in the near future. PMID- 11729751 TI - Genetic modification of plant secondary metabolite pathways using transcriptional regulators. AB - Plant secondary metabolism is the source of many natural products with diverse applications, including pharmaceuticals, food colors, dyes and fragrances. Functions in plants include attraction of pollinating insects and protection against pests and pathogens. An important regulatory step in secondary metabolism is transcription of the biosynthetic genes. The aim of this chapter is to discuss results and opportunities concerning modification of secondary metabolism using transcriptional regulators. The transcriptional regulation of two well-studied secondary pathways, the phenylpropanoid pathway and its flavonoid branch, and the terpenoid indole alkaloid biosynthetic pathway, are reviewed. Some examples of successful engineering of these pathways via transcriptional regulators are discussed. PMID- 11729752 TI - The production of foreign proteins from genetically modified plant cells. AB - While traditionally used to produce natural products, plant suspension cultures can also be utilized for the production of foreign proteins. Production of these high-value products in plant cells is an economically viable alternative to other systems, particularly in cases where the protein must be biologically active. There are several advantages to using plant cells for the large-scale production of secreted proteins. Plant cell media are composed of simple sugars and salts and are therefore less expensive and complex than mammalian media. Consequently, purification of secreted protein is simpler and more economical. Additionally, plant cell derived proteins are likely to be safer than those derived from other systems, since plant cell pathogens are not harmful to humans. In this chapter, we will review foreign protein production from plant cells. To begin, we will discuss the behavior of plant cell cultures, products produced by plant cells, protein secretion and its relationship to purification, and the performance of plant cells as compared to whole plants and other alternative hosts. After a brief discussion of gene transfer techniques, we will present strategies to overcome the limitations of protein production, including protein stabilization, novel production schemes, modeling, and scale-up considerations. To conclude, we will discuss implications for future development of this technology. PMID- 11729753 TI - Large-scale plant micropropagation. AB - Plant micropropagation is an efficient method of propagating disease-free, genetically uniform and massive amounts of plants in vitro. The micropropagation from cells can be achieved by direct organogenesis from hairy roots or regeneration via somatic tissue. Once the availability of embryogenic cell and hairy root systems based on liquid media has been demonstrated, the scale-up of the whole process should be established by an economically feasible technology for their large-scale production in appropriate bioreactors. It is necessary to design a suitable bioreactor configuration that can provide adequate mixing and mass transfer while minimizing the intensity of shear stress and hydrodynamic pressure. Automatic selection of embryogenic calli and regenerated plantlets using an image analysis procedure should be associated with the system. Using the above systems, it will be possible to establish an advanced plant micropropagation system in which the plantlets can be propagated without soil under optimal conditions controlled in plant factory. The aim of this review is to identify the problems related to large-scale plant micropropagation via somatic embryogenesis and hairy roots, and to summarize the most recent developments in bioreactor design. Emphasis is placed on micropropagation technology and computer-aided image analysis, including the successful results obtained in our laboratories. PMID- 11729754 TI - Characterization and application of plant hairy roots endowed with photosynthetic functions. AB - The scope of this review includes the physiological properties of the hairy roots induced by light irradiation and the kinetic analysis considering the effects of light intensity on hairy root cultures. The cell lines of photomixotrophic and photoautotrophic hairy roots of pak-bung are established from heterotrophic ones by improving the photosynthetic ability of hairy roots through acclimation cultures under light irradiation. Comparisons of physiological properties of derived photoautotrophic cell line with photomixotrophic and heterotrophic ones are also made through histological examination. Moreover, the effect of photosynthesis inhibitor on the photoautotrophic growth of the hairy roots is described. By elucidating the influences of light intensity on growth and chlorophyll formation of photoautotrophic and photomixotrophic hairy roots, a kinetic model was applied to describe the hairy root growth and chlorophyll formation of these cell lines of hairy roots. PMID- 11729755 TI - Gas concentration effects on secondary metabolite production by plant cell cultures. AB - One aspect of secondary metabolite production that has been studied relatively infrequently is the effect of gaseous compounds on plant cell behavior. The most influential gases are believed to be oxygen, carbon dioxide and other volatile hormones such as ethylene and methyl jasmonate. Organic compounds of interest include the promising antimalarial artemisinin (known as "qing hao su" in China where it has been a folk remedy for centuries) that is produced by Artemisia annua (sweet wormwood) and taxanes used for anticancer therapy that are produced by species of Taxus (yew). The suspension cultures of both species were grown under a variety of dissolved gas conditions in stoppered culture flasks and under conditions of continuous headspace flushing with known gas mixtures. An analysis is presented to show the culture conditions are such that equilibrium between the culture liquid and gas head-space is assured. The growth rate of the cells and their production rates of artemisinin and paclitaxel were determined. These and other parameters are correlated as functions of the gas concentrations. Interdependence of ethylene and methyl jasmonate is also explored with respect to regulation of secondary metabolite formation. PMID- 11729757 TI - [Approach of noise emission limit value sciences: on the topic of stress during the day and at night]. PMID- 11729756 TI - Integrated bioprocessing for plant cell cultures. AB - Plant cell suspension culture has become the focus of much attention as a tool for the production of secondary metabolites including paclitaxel, a well-known anticancer agent. Recently, it has also been regarded as one of the host systems for the production of recombinant proteins. In order to produce phytochemicals using plant cell cultures, efficient processes must be developed with adequate bioreactor design. Most of the plant secondary metabolites are toxic to cells at the high concentrations required during culture. Therefore, if the product could be removed in situ during culture, productivity might be enhanced due to the alleviation of this toxicity. In situ removal or extractive bioconversion of such products can be performed by in situ extraction with various kinds of organic solvents. In situ adsorption using polymeric resins is another possibility. Using the fact that secondary metabolites are generally hydrophobic, various integrated bioprocessing techniques can be designed not only to lower toxicity, but also to enhance productivity. In this article, in situ extraction, in situ adsorption, utilization of cyclodextrins, and the application of aqueous two-phase systems in plant cell cultures are reviewed. PMID- 11729758 TI - [Reducing individual noise-induced health risks with health psychology and stress coping]. PMID- 11729759 TI - [Specialized assessment by the expert for environmental problems--August 1999]. PMID- 11729760 TI - [Health-related aspects of research on noise effects other than on the ear]. PMID- 11729761 TI - [Analysis of research on noise effects over the past 25 years with reference to the night aircraft noise problem. Reliable knowledge, effects]. PMID- 11729762 TI - [Arousal caused by aircraft noise--environmentally-induced adverse sleep and health effects from the sleep medicine viewpoint]. PMID- 11729763 TI - [The interrupted sleep profile]. PMID- 11729764 TI - [Noise-induced nocturnal cortisol secretion and tolerable aircraft flights]. PMID- 11729765 TI - [Quality goals in preventing detrimental effects and stress caused by aircraft noise]. PMID- 11729766 TI - [Stress hormones and sleep disorders--electrophysiological and hormonal aspects]. PMID- 11729767 TI - [Aircraft noise and circadian rhythm. Possibilities for redistribution and modification of adverse noise effects]. PMID- 11729768 TI - Transformative alliance between qualitative and quantitative approaches in health promotion research. PMID- 11729769 TI - Evaluation of quality-of-life initiatives in health promotion. PMID- 11729770 TI - Economic evaluation of health promotion. PMID- 11729771 TI - Health promotion: towards a quality assurance framework. PMID- 11729772 TI - Investigating policy networks for health: theory and method in a larger organizational perspective. PMID- 11729773 TI - Evaluating community health promotion programmes. PMID- 11729774 TI - Evaluating community initiatives for health and development. PMID- 11729775 TI - Evaluating health promotion in schools: reflections. PMID- 11729776 TI - Evaluating health promotion programmes in the workplace. PMID- 11729777 TI - Evaluation in urban settings: the challenge of Healthy Cities. PMID- 11729778 TI - Evaluating healthy public policies in community and regional contexts. PMID- 11729779 TI - Evaluation of health promotion policies: tracking a moving target. PMID- 11729780 TI - Evaluation of countrywide health promotion policies: the Canadian experience. PMID- 11729781 TI - Health impact assessment as a tool for health promotion and population health. PMID- 11729782 TI - Social capital: evaluation implications for community health promotion. PMID- 11729783 TI - Beyond process and outcome evaluation: a comprehensive approach for evaluating health promotion programmes. PMID- 11729784 TI - Case study: the Canadian Heart Health Initiative. PMID- 11729785 TI - Issues in evaluating mass-media health communication campaigns. PMID- 11729786 TI - Investment for health: developing a multifaceted appraisal approach. PMID- 11729787 TI - Evaluation in health promotion: synthesis and recommendations. PMID- 11729788 TI - What counts as evidence: issues and debates. PMID- 11729789 TI - A framework for health promotion evaluation. PMID- 11729790 TI - Participatory approaches to evaluation in health promotion. PMID- 11729791 TI - [The Lancet supports Danish researchers against the breast cancer group. Mammography screening causes a conflict within Cochrane Collaboration]. PMID- 11729792 TI - [New medical technology--threat or hope? Information about new methods from the SBU Alert]. PMID- 11729793 TI - [The SBU Alert report: Magnetic resonance tomography of knee injuries and in diffuse knee joint problems]. PMID- 11729794 TI - [Big differences when it comes to antibiotic resistance between European intensive care units]. PMID- 11729795 TI - [Ischemic heart disease. Coronary surgery often effective also in diabetes]. AB - The prevalence of diabetes is increasing. These patients are at greater risk of developing ischemic heart disease. Many have symptoms that cannot be adequately managed through pharmacological treatment. Coronary artery bypass grafting (CABG) has shown good effects on survival and quality of life in ischemic heart disease. However, studies on diabetics have been few. Sub-group analyses and observational studies provide indirect support for the contention that CABG increases survival and improves quality of life even among diabetics. When diabetes and multivessel disease coexist, CABG offers advantages as compared with percutaneous transluminal coronary angioplasty (PTCA). CONCLUSION: CABG increases survival and improves quality of life among diabetics with symptoms of ischemic heart disease. PMID- 11729796 TI - [How to manage all those ECGs? Experiences with computer-based storage and interpretation of electrocardiograms in the county of Vastmanland]. AB - We describe our experience using a database (Sicare, Siemens Elema) containing 381,000 ECGs covering 90% of all ECGs during 11 years in the county of Vastmanland, Sweden, with 260,000 inhabitants. The database expedites rapid recovery of earlier ECGs. For instance, in patients admitted to coronary care with chest pain and ST-elevation, speedy access to an earlier ECG with identical ST-elevation may be crucial for avoiding unnecessary and potentially harmful thrombolysis. The database also enables us to select ECGs with a specific diagnosis, for instance atrial fibrillation. This facilitates characterization and quality control of treatment of atrial fibrillation in a well-defined population. PMID- 11729797 TI - [Multilevel analysis of regional disparities in survival after heart failure: differences between county health services affect little patients' prognosis]. AB - The prognosis for patients suffering from heart failure in Sweden differs from county to county, indicating a need for a comparative study in order to uncover the reasons. Conventional single-level analytical methods, however, underestimate the statistical uncertainty in such studies, leading to an inappropriate ranking of regions. Consequently, public opinion and decision-making may be misled. Conventional methods are also unable to disentangle the extent to which differences in prognosis may depend on individual or regional factors. Multilevel analysis, on the other hand, provides a better estimate of statistical uncertainty, and can both identify and quantify the extent to which differences in prognosis depend on either patient-related or regional factors. Using multilevel analysis, we examined the one-year mortality of 38,343 heart failure patients for the period 1992-1995 in every county in Sweden. Regional differences in one-year mortality were found to be very small, the most relevant factors being individual ones. The limited differences distinguishable at the county level may be explained in part by physician density: the more physicians in a county, the better an individual patient's prognosis. PMID- 11729798 TI - [Quality assurance of secondary prevention of coronary artery disease. Ongoing project offers possibilities for improved treatment]. AB - Regional programs for secondary prevention of coronary artery disease have been under development for nearly a decade in Sweden. To achieve maximum adherence these programs were created in close collaboration between hospital and primary care physicians. The programs are local applications of national guidelines and aim to support compliance among both patients and physicians. In January 1998 the Swedish Society of Cardiology and the Swedish Association of General Practice launched a program for quality control and quality assurance of these initiatives. So far, 51 of 79 districts have joined the program. Patients' diaries used for risk factor registration contain 7 report cards on the management of risk factors and medication. These cards are sent to a central registry upon release from the hospital, after 3-6 months, and annually for 5 years. Results from the first year point to differences between the various districts with respect to compliance with both local programs and European guidelines. Overall, results are promising and indicate that this program is successful and leads to improved management of patients with coronary artery disease. PMID- 11729799 TI - [Why, Clara, why?]. PMID- 11729800 TI - [Any place for post-modern theories in medicine?]. PMID- 11729801 TI - [Accident and disaster training of media personnel in hospital environment]. PMID- 11729802 TI - [Dismantling of facilities for mentally disabled is now completed]. PMID- 11729803 TI - [Traumatic stress and suicidal behavior of refugees--epidemiological findings]. PMID- 11729804 TI - [Also "whiplash-related disorders" require evidence-based knowledge]. PMID- 11729805 TI - [A methadone-patient's experiences with the Stockholm program]. PMID- 11729806 TI - [Lipid lowering doesn't affect the development of atherosclerosis in peripheral artery disease]. PMID- 11729807 TI - How important are direct fitness benefits of sexual selection? AB - Females may choose mates based on the expression of secondary sexual characters that signal direct, material fitness benefits or indirect, genetic fitness benefits. Genetic benefits are acquired in the generation subsequent to that in which mate choice is performed, and the maintenance of genetic variation in viability has been considered a theoretical problem. Consequently, the magnitude of indirect benefits has traditionally been considered to be small. Direct fitness benefits can be maintained without consideration of mechanisms sustaining genetic variability, and they have thus been equated with the default benefits acquired by choosy females. There is, however, still debate as to whether or not males should honestly advertise direct benefits such as their willingness to invest in parental care. We use meta-analysis to estimate the magnitude of direct fitness benefits in terms of fertility, fecundity and two measures of paternal care (feeding rate in birds, hatching rate in male guarding ectotherms) based on an extensive literature survey. The mean coefficients of determination weighted by sample size were 6.3%, 2.3%, 1.3% and 23.6%, respectively. This compares to a mean weighted coefficient of determination of 1.5% for genetic viability benefits in studies of sexual selection. Thus, for several fitness components, direct benefits are only slightly more important than indirect ones arising from female choice. Hatching rate in male guarding ectotherms was by far the most important direct fitness component, explaining almost a quarter of the variance. Our analysis also shows that male sexual advertisements do not always reliably signal direct fitness benefits. PMID- 11729808 TI - Planetary habitability: is Earth commonplace in the Milky Way? AB - Is there life beyond planet Earth? This is one of the grand enigmas which humankind tries to solve through scientific research. Recent progress in astronomical measurement techniques has confirmed the existence of a multitude of extra-solar planets. On the other hand, enormous efforts are being made to assess the possibility of life on Mars. All these activities have stimulated several investigations about the habitability of cosmic bodies. The habitable zone (HZ) around a given central star is defined as the region within which an Earth-like planet might enjoy the moderate surface temperatures required for advanced life forms. At present, there are several models determining the HZ. One class of models utilises climate constraints for the existence of liquid water on a planetary surface. Another approach is based on an integrated Earth system analysis that relates the boundaries of the HZ to the limits of photosynthetic processes. Within the latter approach, the evolution of the HZ for our solar system over geological time scales is calculated straightforwardly, and a convenient filter can be constructed that picks the candidates for photosynthesis based life from all the extra-solar planets discovered by novel observational methods. These results can then be used to determine the average number of planets per planetary system that are within the HZ. With the help of a segment of the Drake equation, the number of "Gaias" (i.e. extra-solar terrestrial planets with a globally acting biosphere) is estimated. This leads to the thoroughly educated guess that there should exist half a million Gaias in the Milky Way. PMID- 11729809 TI - Biogeographic methods identify gymnosperm biodiversity hotspots. AB - A remarkable congruence among areas of endemism, panbiogeographic nodes, and refugia in western North America, Japan, south-western China, Tasmania, and New Caledonia indicates that these areas deserve special status for conservation. Here we propose that areas identified by different biogeographic methods are significant candidates for designation as hotspots. PMID- 11729810 TI - Supercooling ability in the house spider, Achaearanea tepidariorum: effect of field-collected and laboratory-reared prey. AB - The influence of prey animals on the supercooling ability of the house spider, Achaearanea tepidariorum, was studied by feeding spiders with field-collected and laboratory-cultured prey animals. Irrespective of the prey species supplied, spiders given field-collected prey had a higher supercooling point than those given laboratory-cultured counterparts. This means that (1) the field prey animals contained some efficient ice nucleators, whereas the laboratory animals were free from such substances and (2) the ice nucleators must be of external origin. Several lines of evidence also suggest that, under natural conditions, potential prey animals for the house spider are highly contaminated with ice nucleators. PMID- 11729811 TI - Female reciprocal calling in the Iberian midwife toad (Alytes cisternasii) varies with male call rate and dominant frequency: implications for sexual selection. AB - Male midwife toads (Alytes cisternasii) responded differently depending on the call frequency and call rate of other males. I tested female Iberian midwife toads with the same set of stimuli used earlier with males. Females responded faster to high call rates, and female vocal activity was greater in response to low-frequency male calls. Thus, in both sexes, the vocal response differs in the same direction according to signal frequency variation, but the magnitude of the response is greater in males than in females. In the light of these results, I discuss the implications for sexual selection of this reciprocal calling. PMID- 11729812 TI - Phonotaxis during walking and flight: are differences in selectivity due to predation pressure? AB - Female selectivity was tested in Tettigonia viridissima during two different phonotaxis situations; compensated walking and tethered flight. For two of the three temporal parameters that are important for call recognition in T. viridissima, selectivity was similar in the two situations. Selectivity for the third parameter (minimum interval duration between the double pulses) was much higher during walking than during flight: walking females responded only to stimuli with intervals of 28 ms or longer, while call models with intervals of 18 ms were attractive during flight. One interneuron (TN-1) is probably involved in filtering the minimum interval duration. As this neuron is also the most likely candidate for transmitting bat calls during flight, it is suggested that the selectivity differences between walking and flying might be due to the need for detecting predator signals during flight, when TN-1 would be occupied listening for bats. With TN-1 unavailable for song processing during flight, temporal selectivity for the minimum interval duration should be reduced, as was found here. PMID- 11729813 TI - Toshiya Yamada. 1960-2001. PMID- 11729814 TI - Pneumococcal conjugate vaccines in Asia and the Pacific. Singapore, 9-11 October 2000. Report of an Asia Pacific Board Advisory Meeting. PMID- 11729815 TI - The extent and effects of abuse and harassment of emergency medicine residents. The SAEM In-service Survey Task Force. AB - OBJECTIVE: To determine the extent and effects of abuse and harassment, including sexual harassment and racial or ethnic discrimination, experienced by residents in emergency medicine (EM). METHOD: The study instrument was an anonymous, self report survey administered to a national sample of EM residents. The survey was timed to coordinate with the American Board of Emergency Medicine's annual In Service Examination in February 1993. The residents reported whether they had experienced nine types of abuse or harassment during their residency training, the sources of these incidents, the effects on the residents, and whether they chose to file a formal complaint regarding these events. RESULTS: Surveys were returned by 1,774 (80%) of the 2,229 residents who sat for the examination--74.4% men and 24.6% women. Overall, 98% reported at least one occurrence of abuse or harassment, with patients being the most frequent source. More than half of the more senior residents reported having been physically hit or pushed. Other health care professionals were a frequent source of verbal abuse and sexual harassment. Women were significantly more likely than men to report unwanted sexual advances (63% vs 32%, p < 0.001), discomfort from sexual humor (66% vs 27%, p < 0.001), and unfair treatment because of gender (71% vs 15%, p < 0.001). Non-Caucasians reported a higher rate of racial or ethnic discrimination than did Caucasians (51% vs 23%, p < 0.001). As a result of these episodes, 19% of the respondents had questioned their decision to become a physician, 20% had questioned entering the specialty of EM, 11% had experienced emotional effects lasting over one month, and 11% had experienced disruption of their family lives. Only 56 (3.2%) had filed formal complaints. CONCLUSIONS: Residents frequently encounter abuse or harassment, particularly from patients. They also report adverse consequences from these episodes. PMID- 11729816 TI - Emergency medicine resident moonlighting: a survey of program directors. CORD Task Force on Resident Moonlighting. AB - OBJECTIVES: 1) To systematically describe emergency medicine (EM) program directors' perceptions of the benefits and risks of resident moonlighting. 2) To assess moonlighting policies of EM residencies, the degree of compliance with these policies, and the methods of dealing with residents who are out of compliance. METHODS: A written survey was mailed or hand-delivered to all allopathic and osteopathic EM residency directors in the United States in 1992 93. Incomplete and ambiguous surveys were completed by phone. RESULTS: There was a 96% response rate (113/118). The average EM resident clinical workweek ranged from 38 to 50 hours while the resident was assigned to ED rotations. Most (90%) of the program directors believe moonlighting interferes with residency duties to some degree. Few (10%) programs prohibit moonlighting altogether, although 44% limit moonlighting to an average of 41.5 hours per month. Program directors believe residents moonlight primarily for financial reasons. Most (60%) of the program directors believe moonlighting offers experience not available in the residency, primarily related to autonomous practice. Fifteen programs reported residents who had been sued for malpractice while moonlighting, with one program director named along with the resident. One third of program directors have penalized residents for abuse of moonlighting privileges. CONCLUSIONS: EM residency directors are concerned about the effect of moonlighting on resident education. The directors' concerns regarding litigation, excessive work hours, and interference with residency duties are balanced by a general acceptance of the financial need to supplement residency income. PMID- 11729817 TI - National survey of emergency medicine resident moonlighting. SAEM Inservice Examination Survey Task Force. AB - OBJECTIVES: To survey emergency medicine (EM) residents regarding moonlighting practices and perceptions for clarifying: 1) resident moonlighting remuneration; 2) any association of perceived educational debt with moonlighting income and hours; and 3) perceptions related to moonlighting (including motivations, impact on resident training, and potential medicolegal difficulties). METHODS: A confidential, voluntary survey was administered to all allopathic EM residents in the United States. This written survey was provided to residents at their in service examinations. Completed forms were anonymously returned by residents or local administrative staff to a central site where all identifiers were removed prior to mailing en mass to the investigators. Comparisons between groups were made using chi-square tests and correlations were assessed using the Pearson correlation coefficient. RESULTS: Seventy-six percent (1,826/2,407) of the surveys were returned. There was a weak correlation (r = 0.11) between educational debt and moonlighting hours for residents in the second year and above, but no association of debt with moonlighting income. Most (88%) of the residents reported that their programs permitted moonlighting. Nearly half (49%) reported that they did moonlight in some way. Most (82%) thought moonlighting provided experience not available in the residency. Only 13 (2%) respondents stated they had been sued for malpractice while moonlighting. Most (66%) moonlighting respondents stated that they moonlighted for financial reasons, with educational debt the primary motivating factor. Of the moonlighting residents, 28% were unsure of their type of malpractice insurance coverage, and 9% had no coverage at all. CONCLUSIONS: Education about EM practice matters including the risks of moonlighting should begin early in residency, because moonlighting is widespread. Residents are vulnerable to medicolegal action while moonlighting and have insufficient knowledge of their malpractice insurance coverage. Although educational debt is perceived as a strong motivating factor for moonlighting, there is only a weak relationship between educational debt and moonlighting hours. PMID- 11729818 TI - [Quality of water of dental units. A survey in the region of Berne, Switzerland]. AB - The aim of this study was to assess the microbial contamination of the water from 175 dental units, which had not been in use for at least 12 hours, and to determine the usefulness of rinsing the water lines for 3 minutes. Only 10% of all units fulfilled all criteria for drinking water. Frequently the total bacterial counts markedly exceeded 100 CFU/ml. In all cases the rinsing decreased bacterial loads significantly. Bacterial contamination of water lines increased with the time the dental units remained unused. It is recommended to rinse dental unit water lines daily for at least 3 minutes before working on patients. PMID- 11729819 TI - [In vitro comparison of an electronic root canal length measuring device and the radiographic determination of working length]. AB - The objective of this study was to evaluate the accuracy of an electronic root canal length measuring device (AFA Apex Finder, Analytic Technology, Orange, USA) and to compare the results with those obtained using handfiles evaluated radiographically using 146 extracted teeth with mature single roots. Root canal lengths were determined electronically with files cemented to their lengths. A radiograph in long cone technique with the inserted root canal instrument was performed. The apical point of working length was determined 1 mm coronally of the radiographic apex. The apical part of root surfaces was removed to expose the root canal with the inserted probe tip. The distance between measuring file tip and the apical constriction was measured using a microscope. In 77.2% of the evaluated radiographs the determined apical point of working length was found in a range of +-0.5 mm from the apical constriction. In 98.5% of the results of electronic determination of working length, the distance between file tip and the apical constriction was less than 0.5 mm. The electronic root canal length measuring device provided significantly more accurate results than the radiographs. PMID- 11729820 TI - Six hospitals and health systems make list of "100 best companies for working mothers". PMID- 11729821 TI - [Capillary microscopy in connective tissue diseases]. PMID- 11729822 TI - [Premedication before endoscopy]. PMID- 11729823 TI - Reduce risks of patients who leave too soon. AB - Patients who leave without being seen present many risks for patient outcomes and liability. Don't assume that patients who leave do not have life-threatening conditions. Call back patients to ask them to return, and see them quickly if they do. Keep statistics on patients who leave to show to accreditation surveyors and to use when developing budgets for the ED. PMID- 11729824 TI - Superantigens and their role in autoimmune disorders. AB - The ability of superantigens to activate large numbers of T cells suggests that they may play a role in the course of autoimmune disorders. Data from several animal models of autoimmune disorders including experimental allergic encephalomyelitis and collagen induced arthritis supports this hypothesis. Administration of bacterial superantigens can induce an exacerbation of the autoimmune process in these models, or induce disease de novo in the appropriately immunized animal. Studies of several human disorders including rheumatoid arthritis, Kawasaki disease, insulin-dependent diabetes, and psoriasis lend credence to the concept that bacterial superantigens may play a role in the pathogenesis of these diseases. Nevertheless, in some cases, depending on the timing of administration and the model, superantigens may lead to an amelioration of the autoimmune process. Based on these results in seems logical to conclude that superantigens can have a significant impact on the course of the immune and autoimmune mediated disorders. PMID- 11729825 TI - Koch's postulates and autoimmunity: an opposing viewpoint. AB - Autoimmunity is characterized as a state of abnormal specific humoral and cell mediated responses against constituents of body tissues. One time-honored approach to explaining the pathogenesis of autoimmunity has been application of the Koch's postulates, on loan from the field of microbiology suggesting that autoantibodies and/or autoreactive T cells are the presumed "pathogens" of autoimmunity, and that passive transfer of these autoimmune factors to susceptible animals will result in the induction of the autoimmune disease. We suggest that autoimmunity is not in many cases due to the presence of factors leading to the autoimmune response in those susceptible. Instead, it is the lack of a factor which leads to the development of autoimmunity, a factor (cytokine, protein, gene, etc.) which is present in the healthy individual and normally protects in from disordered immune regulation. We propose to direct more research into therapeutic modulation of autoimmunity by administration of putative "protective factors", rather than by attempts to depress or remove autoreactive cells and antibodies from the autoimmune. PMID- 11729826 TI - Gene therapy for autoimmune demyelinating disease of the central nervous system. AB - Gene therapy is currently being explored as a new therapeutic treatment of autoimmune disease. The genetic modification of autoreactive memory T cells (T cell-mediated gene therapy) and autoimmune target tissue (target tissue gene therapy) to produce immunoregulatory cytokines offers a promising way to regulate autoimmunity. Furthermore, regenerative gene therapy offers the possibility of delivering growth factors to damaged autoimmune target tissue as a way of mediating repair. In the current review we discuss the different experimental models that are being used to test the efficacy of gene therapy in that treatment of autoimmune disease. We also discuss the importance of regulating transgene expression to ensure the therapeutic transgene products are delivered specifically to the autoimmune milieu in an antigen-inducible, non-constitutive manner. PMID- 11729827 TI - Decrease of enhanced interferon alpha levels in sera of HIV-infected and AIDS patients receiving combined antiretroviral therapy. AB - In the advanced stages of human immunodeficiency virus (HIV) infection the defective interferon (IFN) responses have been observed. Persisting high lebels of the acid-labile interferons (al-IFNs) have been found in sera of the patients with AIDS. The combined antiretroviral therapy, that included the reverse transcriptase and viral protease inhibitors, resulted in a significant improvement of the clinical state of the majority of HIV-infected patients. In this report we describe the levels of IFNs in 41 HIV patients subjected to the combined treatment. High IFN levels (median 84, up to 576 U/ml) were found in sera of patients classified as the stage C2 or C3 of AIDS before the treatment. The combined therapy resulted in the decrease of IFN levels (median 7.5, up to 24 U/ml) that approached the levels of IFNs detected in the HIV+, A1-A3 stage patients (median 4, up to 36 U/ml). In contrast, the unsuccessful therapy connected with the worsening of the clinical state and the decrease of CD4+ cell count had no effect on the IFNs levels (median 48, up to 96 U/ml). Thus, the measurements of IFN activity in sera may be useful for monitoring the effects of the antiretroviral combined therapy. In sera of the AIDS patients, subjected to the antiviral bioassays, the mixture of the acid-labile and acid-stable form of IFN-alpha, with the prevailing al-IFN-alpha, have been detected. PMID- 11729828 TI - In vitro secretion of interleukin 2 and expression of IL-2 receptor in peripheral blood lymphocytes in high risk of insulin-dependent diabetes mellitus subjects. AB - Interleukin 2 (IL-2)--a Th1 lymphocyte-derived cytokine is at present considered to play an important role in the etiopathogenesis of insulin-dependent diabetes mellitus. In the previous studies increased, decreased and unchanged IL-2 levels in patients with recent onset of insulin-dependent diabetes mellitus (IDDM) were found. These differences could be a result of different metabolic status or/and a different stage of the autoimmune process. The aim of our study was to estimate in vitro secretion of IL-2 and CD25 antigen expression by the peripheral blood T lymphocytes in subjects at the preclinical stage of IDDM (prediabetes), but still without metabolic disturbances. In 27 first degree relatives of IDDM patients with antibodies against different pancreatic islet cell antigens (ICA, GADA, IAA, IA-2) CD25 antigen expression on peripheral blood lymphocytes T was measured by flow cytometry and IL-2 concentration in supernatants of 48 and 72 h cultures of peripheral whole blood with 10 microg/ml PHA was estimated by ELISA. The control group was comprised of 34 age and sex-matched healthy volunteers. In the studied high risk IDDM subjects the decreased CD25 expression in peripheral CD4+ lymphocytes T and a negative correlation between the percentage of CD25+ cells and islet cell antibodies (ICA) titres was observed. No differences in IL-2 levels in supernatants of 48 h and 72 h blood cultures was found in subjects with single antibody (ICA+) in comparison to healthy controls. A significant increase of IL-2 secretion at 72 h of PHA stimulation was shown in first degree relatives of IDDM patients with a combination of 3 or more antipancreatic-B cell antibodies. There were also a significant negative correlation between glutamic acid decarboxylase antibodies (GADA) titres and IL-2 levels in 72 h of culture. The present study suggests the involvement of IL-2 in the pathogenesis of IDDM. The estimation of CD25 antigen expression in the peripheral blood lymphocytes could be an additional immunological marker of identification of subjects in prediabetes. PMID- 11729829 TI - Capability of adriamycin and busulfan to induce adaptive response in vitro. AB - The capability to induce an adaptive response by low doses of busulfan (BS) or adriamycin (ADR) was studied in two kinds of mammalian cells with acquired or inherent resistance to ADR (ME18/R and V3) cultured in vitro. The results indicate the presence of an adaptive response to ADR in both kinds of used cells pretreated with a low priming dose of ADR. In the same kind of cells no adaptive response to BS after priming with a low dose of this drug was found. PMID- 11729830 TI - HLA-A, B, C antigens in pulmonary sarcoidosis in Polish population. AB - The aim of this study was to analyze association between HLA class I antigens and sarcoidosis in Poland. HLA-A, B, C antigens in a group of 100 patients suffering from sarcoidosis and in a group of 100 healthy blood donors were determined. Histocompatibility typing was performed by the NIH method using commercially available sera. For statistical analysis chi2 test was used after Yates' correction. The relative risk was calculated by Woolf's method. We found that HLA B8 and -Cw7 prevalence was significantly higher in patients with sarcoidosis than in healthy controls. HLA-B35, -B40, -Cw2 and -Cw4 antigen expression was significantly lower in pulmonary sarcoidosis than in the tested group of healthy individuals. The highest relative risk of sarcoidosis was connected with HLA-B8 and -Cw7. The results obtained suggest that, in the population suffering from pulmonary sarcoidosis in nothern Poland, as compared with the control group of healthy persons, antigens HLA-B8 and -Cw7 are significantly more frequent. It can be assumed that, the presence of these antigens may be connected with a greater risk of pulmonary sarcoidosis. In the group of patients, as compared with the control population, the occurrence of antigens HLA-B35, -B40, -Cw2 and -Cw4 is significantly more rare. PMID- 11729831 TI - Effect of granulocyte colony stimulating factor treatment on ex vivo cytokine production by blood cells of patients after chemotherapy or radiotherapy. AB - We explored ex vivo alterations in the cytokine release of stimulated blood cells taken from 8 patients with hematological malignancies who, after chemotherapy or radiotherapy developed leukopenia, and were treated for 3-7 days subcutaneously with granulocyte colony stimulating factor (G-CSF), daily, dose of 5 microg/kg of body weight. Blood was also taken from 8 healthy controls not treated with G-CSF and from patients before and 24 h after last dose of G-CSF and ex vivo treated with interferon (IFN) inducers: Newcastle disease virus (NDV), phytohemagglutinin (PHA), concanavalin A (Con A) and with tumor necrosis factor (TNF) inducer- lipopolysaccharide (LPS). Blood cells of patients before G-CSF treatment exhibited ex vivo a low ability to produce IFN-gamma in comparison to controls. After G-CSF therapy a significant increase in IFN-alpha production ability was detected. We conclude that G-CSF treatment for 3-7 days does not only increase the number of white blood cells (WBC) and neutrophilic granulocytes but also modify the host response of patients with hematological malignancies to microbial infections. PMID- 11729832 TI - The immunological basis of current and novel therapies of multiple sclerosis. AB - The etiology and the pathogenesis of multiple sclerosis are not yet known. There might be a role for genetic susceptibility, for environmental factors and for inflammatory and immunological changes. Most of the actual therapies are based on the latter two phenomena. We review here corticosteroids, interferon beta and copolymer 1 as the current drugs of choice and compare schematically the immunological basis of the mechanisms of action of these three substances with those of experimental or other treatments. PMID- 11729833 TI - An important previously unpublished manuscript by A. M. Kellas. PMID- 11729834 TI - Bernhard Schultze and the swinging neonate. PMID- 11729835 TI - [13C]linoleic acid oxidation and transfer into milk in stunted lactating women with contrasting body mass indexes. AB - BACKGROUND: The fat concentration of human milk is associated with maternal adiposity, but there is no clear understanding of the mechanisms controlling milk fat concentration. OBJECTIVE: We evaluated the effect of postpartum body mass index (BMI; in kg/m(2)) on the metabolic distribution of an oral dose of [13C]linoleic acid in lactating women. DESIGN: Ten lactating women stratified by BMI (either <22.5 or >23.5) at 5 mo postpartum received orally 2.5 mg [13C]linoleic acid/kg body wt. Exhaled air, milk, and plasma samples were collected in relation to tracer administration. Linoleic acid was determined by gas chromatography. Dietary intake, serum, milk composition, [13C]linoleic acid enrichment in milk and plasma, and exhaled 13CO2 (by isotope ratio mass spectrometry) were assessed. RESULTS: Women with a higher BMI exhaled more 13CO2 than did women with a lower BMI (22.8 +/- 9.4% compared with 8.6 +/- 3.5% of dose, P < 0.03). Cumulated 72-h transfer of [13C]linoleic acid to milk was not significantly different between groups (14.8 +/- 6.5% compared with 17.7 +/- 6.7% of dose). Within the first 9 h after dose administration, 51.6 +/- 4.9% of the total isotope transfer into milk had passed in women with a higher BMI, but only 24.0 +/- 15.3% had passed in those with a lower BMI (P = 0.02). CONCLUSIONS: Women with a lower BMI, who were reputed as having less body fat, oxidized and secreted into milk less dietary linoleic acid within 12 h after tracer administration than did women with a higher BMI. In both groups, a large proportion of [13C]linoleic was retained in the maternal compartment, most likely fat tissue, in a slow turnover pool, and released slowly in later hours. PMID- 11729836 TI - Dopamine dose and renal damage. PMID- 11729837 TI - Propofol use in head-injury patients. PMID- 11729838 TI - Premature mortality and hypopituitarism. PMID- 11729839 TI - Premature mortality and hypopituitarism. PMID- 11729840 TI - Normalisation of blood pressure in bleeding patients. PMID- 11729841 TI - Causes of obesity. PMID- 11729842 TI - Inhaled insulin in type 1 diabetes. PMID- 11729843 TI - Inhaled insulin in type 1 diabetes. PMID- 11729844 TI - Inhaled insulin in type 1 diabetes. PMID- 11729845 TI - Repeal of the Boren amendment: potential implications for long-term care. PMID- 11729846 TI - Improving coverage for low-income Medicare beneficiaries. PMID- 11729847 TI - Assessing quality in managed care: health plan reporting of HEDIS performance measures. PMID- 11729848 TI - Community health centers in a changing U.S. health care system. PMID- 11729849 TI - The President's proposed long-term care initiative: background and issues. PMID- 11729850 TI - Designing a Medicare drug benefit: whose needs will be met? PMID- 11729851 TI - Effect of phosphorus availability on basal root shallowness in common bean. AB - Root gravitropism may be an important element of plant response to phosphorus availability because it determines root foraging in fertile topsoil horizons, and thereby phosphorus acquisition. In this study we seek to test this hypothesis in both two dimensional paper growth pouch and three-dimensional solid media of sand and soil cultures. Five common bean ( Phaseolus vulgaris L.) genotypes with contrasting adaptation to low phosphorus availability were evaluated in growth pouches over 6 days of growth, and in sand culture and soil culture over 4 weeks of growth. In all three media, phosphorus availability regulated the gravitropic response of basal roots in a genotype-dependent manner. In pouches, sand, and soil, the phosphorus-inefficient genotype DOR 364 had deeper roots with phosphorus stress, whereas the phosphorus-efficient genotype G19833 responded to phosphorus stress by producing shallower roots. Genotypes were most responsive to phosphorus stress in sand culture, where relative root allocation to the 0-3- and 3-6-cm horizons increased 50% with phosphorus stress, and varied 300% (3-6 cm) to 500% (0-3 cm) among genotypes. Our results indicate that (1) phosphorus availability regulates root gravitropic growth in both paper and solid media, (2) responses observed in young seedlings continue throughout vegetative growth, (3) the response of root gravitropism to phosphorus availability varies among genotypes, and (4) genotypic adaptation to low phosphorus availability is correlated with the ability to allocate roots to shallow soil horizons under phosphorus stress. PMID- 11729852 TI - From tobacco to health care and beyond--a critique of lawsuits targeting unpopular industries. AB - The 1998 settlement between state Medicaid agencies and the five major tobacco companies heralded a new form of litigation in which individual or government plaintiffs allied with private class action attorneys use economic, political and moral leverage to extract huge settlements from entire industries. Beginning with several class action suits filed in late 1999 against managed care companies by aggrieved HMO enrollees, and continuing with government suits against the paint and handgun industries, this new form of litigation has become a powerful vehicle for plaintiffs to punish unpopular--but entirely legal--industries. In this Note, the author demonstrates that the popular appeal of these suits conceals legal theories of recovery that probably could not survive courtroom scrutiny. The author argues that the thin legal merits of these class action claims are often tolerated by courts, who urge settlement in order to clear their dockets, and by the industries, who regard settlement merely as a cost of doing business. The author concludes that the tobacco litigation and its progeny encourage citizens and the executive branches of government to seek restitution and fundamental social change in the courts after losing in the legislative arena, thus forcing the judiciary branches into the unwise and improper role of policymaker. PMID- 11729853 TI - In vitro and in vivo matrix metalloproteinase production by pancreatic cancer cells and by distant organs. AB - BACKGROUND: Matrix metalloproteinases (MMP), especially MMP-2 and MMP-9, are thought to play major roles in pancreatic cancer growth and metastasis. MMP inhibition prolongs survival in a murine model of human pancreatic cancer. To establish an in vitro model of human pancreatic cancer, we sought to determine in vitro MMP production by two human pancreatic cancer cell lines, compare in vitro to in vivo production, and, if different, determine if in vitro MMP production could be made to mimic in vivo production by altering the in vitro environment of the cell lines. We also sought to document that in vivo production of MMP was unique to orthotopic pancreatic cancer cells. METHODS: HPAC and PANC-1 cell lines were grown in vitro and in a murine model of orthotopic pancreatic cancer. MMP-2 and MMP-9 production by the cell lines was determined. Noting differences between in vitro and in vivo production, the cell lines were stimulated in vitro with PMA. Orthotopic pancreatic tumor MMP production was compared to splenic, hepatic, and renal production of MMP. MMP-2 and MMP-9 production was sought in both active and latent forms utilizing gelatin zymography. Results were confirmed by Western blot. Densitometry allowed comparisons: scale 0 (none) to ++++ (most). RESULTS: In vitro, HPAC and PANC-1 produce modest amounts of MMP-2 and MMP-9, mostly in latent form. In vivo, both cell lines produce more MMP, particularly in the active forms. In vitro stimulation of the cell lines with the PMA increased MMP production, particularly of active MMP-2 and latent MMP-9, resulting in a profile of MMP production similar to that seen in vivo. Hepatic, renal and splenic MMP levels, especially active forms, were very low relative to the tumors. CONCLUSIONS: Pancreatic cancer cell lines produce less MMP, especially active form, in vitro than in vivo. With in vitro stimulation, the MMP profile of cell lines mimic those of tumors in vivo, suggesting that cancer cells are stimulated in vivo. Both in vitro and in vivo, the poorly differentiated cell line PANC-1 is associated with broadened and enhanced MMP activity. Healthy solid organs produce little MMP-2 or MMP-9, most of which is latent form. PMID- 11729854 TI - Role of cytoplasmic and secretory phospholipase A2 in intestinal epithelial cell prostaglandin E2 formation. AB - BACKGROUND: Prostanoid production is dependent on the enzymatic activity of phospholipase A2 enzymes to produce the precursor, arachidonic acid. Two principle phospholipase A2 enzymes play a major role in arachidonic acid production, 85kDa cytoplasmic phospholipase A2 (cPLA2) and 14kDa secretory phospholipase A2 (sPLA2). The purpose of this study was to determine the PLA2 enzyme involved in prostanoid formation in intestinal epithelial cells. METHODS: Employing a human and murine intestinal epithelial cell line, cells were exposed to the stimulants lipopolysaccharide (LPS), interleukin 1beta (IL-1) and calcium ionophore (Ca Ion) in the presence and absence of cPLA2 and sPLA2 inhibitors. The expression of both PLA2 enzymes and prostaglandin E2 (PGE2) formation were determined. RESULTS: Western blotting demonstrated that the cPLA2 enzyme was constitutively expressed in the human cell lines and not evidently increased by exposure to any of the stimulants. In murine cells the cPLA2 enzyme was also constitutively expressed and not induced by the stimulants evaluated. The sPLA2 enzyme was constitutively expressed in both cell lines and appeared to be induced by LPS and IL-1 in human enterocytes but not by Ca Ion. In murine enterocytes sPLA2 was induced by all three stimuli. PGE2 production by the human cell line was increased by LPS, IL-1 and Ca Ion. IL-1 and Ca Ion stimulated PGE2 formation was inhibited by the cPLA2 enzyme inhibitors while LPS stimulated PGE2 production was not inhibited by the cPLA2 inhibitor; but was inhibited by the sPLA2 enzyme inhibitor. Murine epithelial cells increased PGE2 formation in response to IL-1 and Ca Ion, but not LPS and the increased PGE2 was significantly decreased by cPLA2 enzyme inhibitors. CONCLUSIONS: The metabolic pathway of PGE2 formation is variable and the PLA2 enzyme involved in producing PGE2 is dependent on the stimulus and the cell line. In human intestinal epithelial cells, LPS production of PGE2 proceeds through a pathway associated with sPLA2 generated arachidonic acid while IL-1 stimulated PGE2 is produced by arachidonic acid generated by cPLA2. The physiologic significance of the various metabolic pathways of PGE2 formation is unknown. PMID- 11729855 TI - Enhanced arterial contractile responses in diabetic hypercholesterolemic pig carotid arteries. AB - BACKGROUND: Two of the common risk factors for atherosclerosis are diabetes and hypercholesterolemia. AIMS: The purpose of this investigation was to determine the independent and combined effects of experimental diabetes (DM) and hypercholesterolemia (HC) on vasomotor responses. METHODS: Yorkshire pigs were rendered hypercholesterolemic, diabetic, hypercholesterolemic/diabetic, or maintained as control. Dose dependent physiologic responses of the carotid artery smooth muscle to serotonin, sodium nitroprusside and phorbol ester were determined in a muscle bath. RESULTS: Vascular smooth muscle from the DM and HC/DM pigs showed enhanced contractile responses to serotonin. The HC pigs showed diminished contractile responses to serotonin. The DM smooth muscles had enhanced contractile responses to phorbol dibutyrate. There were no differences in endothelial-independent relaxation responses to sodium nitroprusside. CONCLUSIONS: This study demonstrates that diabetes enhances smooth muscle contractile responses in the presence and absence of hypercholesterolemia. PMID- 11729856 TI - C-erbB-2 oncoprotein content in colorectal cancer and in surrounding mucosa: relationship with clinicopathologic parameters and prognostic significance. AB - BACKGROUND: c-erbB-2 is a transmembrane signaling molecule closely related in structure to the epidermal-growth-factor receptor (EGFR) but biologically distinct from it. c-erbB-2 has been implicated in cell transformation and tumor pathogenesis, but very little is known about its content and clinical significance in colorectal cancer. AIMS: To evaluate the c-erbB-2 content in colorectal cancer and its possible relationship with clinicopathologic parameters from tumors and prognostic significance. METHODS: Membranous and cytologic c-erbB 2 oncoprotein contents were examined by an immunoenzymatic assay in tumors and paired normal surrounding mucosa samples from 131 colorectal cancer patients. In addition, survival analysis were prospectively performed in a subgroup of 69 consecutive patients with resectable colorectal carcinomas, who underwent a mean follow-up period of 28 months. RESULTS: In the overall group of patients, c-erbB 2 levels were significantly higher in membranous than in cytosolic samples, in neoplastic tissues (5,830.4 +/- 1085.3 vs. 934.2 +/- 107.5 NHU/mg protein; p < 0.0001) and in surrounding normal mucosa samples (5,257.8 +/- 646.3 vs. 837.4 +/- 187.4 NHU/mg protein; p < 0.0001). Nevertheless, a significant positive relation was found between membranous and cytosolic oncoprotein levels in these two paired sets (p < 0.0001, for both). There were no significant differences in membranous or cytosolic c-erbB-2 protein levels between neoplastic tissues and surrounding mucosa samples in this overall group of patients. In addition, the results did not show significant correlations of these oncoprotein contents with clinicopathologic parameters from tumors such as location, stage, histologic grade, and DNA content or S-phase fraction. However, the results indicated that low membranous c-erbB-2 content (< 4,500 NHU/mg protein) in tumors predict shorter relapse-free survival and overall survival (p < 0.05, for both) in resectable colorectal cancer patients. CONCLUSIONS: There are a wide variability of both membranous and cytologic c-erbB-2 contents in colorectal carcinomas, which seems to correspond to the biological heterogeneity of these tumors. In addition, our results also demonstrate that high membranous c-erbB-2 levels are associated with lesions of favorable evolution in resectable colorectal cancer patients. PMID- 11729857 TI - Effect of age on some blood variables relating to bone metabolism in women. AB - BACKGROUND: The available laboratory parameters reflecting bone metabolism are not adequate for reliable diagnosis of osteoporosis. They display a marked biological variation and are inaccurate in individual cases. Therefore precise knowledge of these variations, as upon aging in healthy people is important. AIMS: The purpose was to examine the age- and weight-related variation of some blood constituents relating to mineral metabolism and commonly used in hospital laboratories in healthy women aged over 40, and to estimate their mutual correlations and normal values in different age groups. METHODS: The study series consisted of 238 healthy Caucasian women without any diseases related to bone metabolism and aged 40-86 years, who were divided into the following age categories: 40-45, 50-55, 60-65, 70-75 and over 80 years. RESULTS: Markers of bone formation, alkaline phosphatase (AP) and osteocalcin, and marker of bone resorption, tartrate resistant acid phosphatase (TrAcP), as well as parathyroid hormone, phosphorus and creatinine increased with age, whereas 25-hydroxyvitamin D and oestradiol decreased. All these parameters except calcium showed a significant age relation. Only the relation of weight versus osteocalcin and weight versus TrAcP remained significant when the effect of age was included in the multiple regression analysis or the partial correlation coefficients were examined. There was a significant correlation between serum osteocalcin and serum AP. Serum TrAcP had a significant positive correlation with serum osteocalcin and serum AP. Percentage fat mass correlated significantly with AP and TrAcP. CONCLUSION: Our observations may be useful when these markers of bone metabolism having inadequate sensitivity and specificity, are used as a battery in the diagnosis of osteoporosis and other metabolic bone diseases and in the assessment of normality in population studies. PMID- 11729858 TI - Laparoscopic versus open adrenalectomy: outcome in 35 consecutive patients. AB - BACKGROUND: The use of minimally invasive techniques in the surgical treatment of adrenal masses has been used to remove a wide variety of adrenal tumors. AIMS: We have reviewed our experience with laparoscopic adrenalectomy and compared laparoscopic vs. open surgical approach. METHODS: The outcome of 35 consecutive patients who underwent adrenalectomy over a 3-year period has been analyzed retrospectively. Differences in operating time, blood loss, period of hospitalization, use of parenteral analgesia, resumption of oral feeding, complications, and time to return to normal activity after 18 coelioscopic vs. 17 open consecutive adrenalectomies have been considered. RESULTS: The average operative time was longer (mean 160 vs. 148 min, p = 0.48) and postoperative complications lower (4 vs. 5 cases, p =0.73), although not statistically significant, for the laparoscopic compared to the open surgical approach, whereas blood loss (30 vs. 165 ml; p = 0.01), postoperative analgesia (3.4 vs. 5.0 days, p = 0.02), time to restart oral feeding (3.0 vs. 4.7 days, p = 0.001), average time of hospitalization (4.5 vs. 9.6 days, p = 0.001), time to return to normal activity (21 vs. 37 days, p = 0.001) were all statistically significant. CONCLUSIONS: Laparoscopic adrenalectomy can be considered the method of choice for managing almost all adrenal masses, because of its lower morbidity and shorter postoperative recovery. PMID- 11729859 TI - The association between telomerase activity and hormone receptor status and p53 expression in breast cancer. AB - BACKGROUND: Telomerase is a ribonucleoprotein enzyme that seems to play an important role in cellular immortality and carcinogenesis. p53 mutations account for approximately 50% of human cancers and represent the most frequent genetic lesion in breast cancer. AIMS: This study aims to examine the association between telomerase reactivation and hormonal receptor status and p53 expression in invasive breast cancer. METHODS: Using a polymerase chain reaction-based assay, telomerase activity was determined in 47 invasive breast carcinomas and 21 adjacent non-cancerous breast tissue specimens (stored at -80 degrees C) prospectively collected from 47 women undergoing elective surgical treatment in our centre. The histopathological features of the tumour were determined by experienced breast pathologists using light microscopy and haematoxylin and eosin staining. Oestrogen receptor (ER), progesterone receptor (PR) and p53 expressions were determined using immunohistochemistry techniques. RESULTS: Telomerase activity was detected in 34 (72%) of 47 breast carcinomas and in none of the adjacent non-cancerous breast specimens. There was a significant association between telomerase reactivation, tumour size and nodal status. Telomerase positive tumours were more likely to be poorly differentiated (65% versus 46%), but this association failed to reach statistical significance. There was no significant difference in ER expression (68% versus 85%). PR expression (62% versus 62%) and p53 expression (19% versus 27%) between telomerase positive and telomerase negative cancers. CONCLUSION: Telomerase reactivation is associated with important prognostic factors such as tumour size and nodal status in invasive breast cancer and seems to be independent of hormonal receptor status and p53 expression. PMID- 11729860 TI - Location of tumor and distribution of lymph node metastasis in gastric cancer: lesser curve or greater curve. AB - BACKGROUND: The aim of this study was to clarify the relationship between the location of tumor and distribution of lymph node metastasis in node-positive gastric cancer. Pathologic findings of tumors in which location of tumors or positive nodes was confined within the lesser or greater curve were evaluated, and candidates of limited resection were assessed. METHODS: A total of 211 patients with node-positive gastric cancer treated by D2 or D3 gastrectomy was studied. Location, size, gross type, and histologic type of tumors, presence or absence of serosal, lymphatic, and vascular invasions, and status of lymph node metastasis were investigated. Tumors were divided into lesser curve tumors, greater curve tumors, and others; and lymph node metastases were classified as lesser curve metastasis, greater curve metastasis, and both curves metastasis. RESULTS: Lesser or greater curve tumors were featured by small size, negative serosal invasion, negative vascular invasion, metastasis confined to the perigastric nodes, and number of positive nodes less than seven. Cases of lesser or greater curve metastasis were featured by tumor location within the lesser or greater curve, small size, grossly superficial type, negative serosal invasion, negative lymphatic invasion, and number of positive nodes less than seven. When size of lesser or greater curve tumors was less than 4 cm, lymph node metastasis was mostly restricted to the same lesser or greater curve. CONCLUSION: Among node positive gastric cancers, tumors located within the lesser or greater curve were characterized by small size, negative serosal invasion, and a few metastases confined to the perigastric lymph nodes. When tumor size was less than 4 cm and tumor invasion was not into serosa, lesser or greater curve tumors rarely extend into lymph nodes along the other curve. Limited resection or less invasive treatment is acceptable for such small gastric cancers along the lesser or greater curve. PMID- 11729861 TI - Surgical treatment of non-ampullary duodenal cancer: good long term survival after radical tumour resection including lymphadenectomy. AB - AIMS: The aim of this study is to evaluate the role of surgery in the treatment of duodenal cancer. METHODS: From October 1987 to January 1999, 7 patients (3 female/4 male) with primary non-ampullary duodenal adenocarcinoma underwent surgical treatment in our department. Presenting signs and symptoms were abdominal pain, weight loss, anaemia and obstruction. Upper gastrointestinal contrast study, computed tomography (CT) and duodenoscopy were the primary diagnostic procedure modalities. All diagnoses were confirmed histologically. Five of 7 patients were resectable for cure at the time of their first surgical intervention. RESULTS: In 5 patients, the tumour could be removed with tumour free margins. One patient, where the histopathological workup revealed tumour cells in the resection margins, died 29 months after the surgical intervention. In another patient, complete tumour resection could not be achieved. Subsequently, postoperative survival was only 2 months. Operative mortality was 0%. Patients with R0-status showed a postoperative survival of at least 24 months. CONCLUSION: In the management of resectable non-ampullary malignancies of the duodenum, surgical radicality including lymphadenectomy should be pursued. The radical operative treatment with lymphadenectomy offers a reasonable chance for cure in some patients. PMID- 11729862 TI - Respiratory control of hypoglossal motoneurons. PMID- 11729863 TI - Projections from brainstem GABAergic neurons to the phrenic nucleus. PMID- 11729864 TI - Optical recording of the neuronal activity in the brainstem-spinal cord: application of a voltage-sensitive dye. AB - Although there are several limitations, optical recording techniques are superior to multi-electrode mapping methods, as it is possible to record at large number of points in a small area without destroying the tissue and possible to know relative changes of membrane potentials. Optical recording techniques using voltage-sensitive dyes will be more importantly applied in the study of central respiratory control (e.g., mechanisms of respiratory rhythm generation) in the near future. PMID- 11729865 TI - Interfacing computer models with real neurons: respiratory "cyberneurons" created with the dynamic clamp. PMID- 11729867 TI - Unstable breathing rhythms and quasiperiodicity in the pre-Botzinger complex. PMID- 11729866 TI - Is the vertebrate respiratory central pattern generator conserved? Insights from in-vitro and in-vivo amphibian models. PMID- 11729868 TI - Stationary organotypic culture of the pre-Botzinger complex from the newborn rat. PMID- 11729869 TI - Respiratory rhythm generation: preBotzinger neuron discharge patterns and persistent sodium current. AB - Considerable evidence from several laboratories (c.f., Rekling and Feldman, Ramirez et al.) is consistent with the concept that the pBc contains the kernel of the central rhythm generating network for breathing. The work summarized in this manuscript is also generally consistent with this notion. Of particular note is the observation that pre-I neurons and E-Dec neurons maintain a consistent phase relationship with phrenic nerve activity and maintain a similar peak discharge rate despite marked changes in the phrenic nerve rhythm and pattern. Other categories of respiratory neurons failed to maintain this relationship. Hence, the findings are consistent with pBc pre-I and E-Dec neurons having a key role in rhythm generation. A persistent sodium current has been postulated to underlie the rhythm generating mechanism of pacemaker neurons within the pBc. In the present study, a substantial persistent sodium current was documented in many neurons from the pBc and adjacent respiratory regions. This finding is not inconsistent with the postulated role in rhythm generation. However, it does suggest that other neuronal properties must act in concert with the persistent current to define a unique population of pacemaker neurons. PMID- 11729870 TI - Roles of the Botzinger complex in the formation of respiratory rhythm. PMID- 11729871 TI - Models of neuronal bursting behavior: implications for in-vivo versus in-vitro respiratory rhythmogenesis. PMID- 11729872 TI - Neurogenesis of the respiratory pattern: insights from computational modeling. PMID- 11729873 TI - Central respiratory chemosensitivity: cellular and network mechanisms. PMID- 11729874 TI - Reconfiguration of the central respiratory network under normoxic and hypoxic conditions. PMID- 11729875 TI - How is the respiratory central pattern generator configured and reconfigured? A workshop summary. PMID- 11729876 TI - Respiratory suppression induced by nicotine withdrawal in the neonatal rat brainstem: implications in the SIDS risk factor. PMID- 11729877 TI - Effect of DNA hypomethylation on neural control of respiration: a genetic model. PMID- 11729878 TI - Respiratory pattern and hypoxic ventilatory response in mice functionally lacking alpha2A-adrenergic receptors. PMID- 11729879 TI - Do genetic factors influence the Dejours phenomenon? PMID- 11729880 TI - Role of nitric oxide in short-term potentiation and long-term facilitation: involvement of NO in breathing stability. PMID- 11729881 TI - Induction of long-term modulation of the exercise ventilatory response in man. PMID- 11729882 TI - Metabolic consequences of hypoxic conditioning in Lymnaea stagnalis. PMID- 11729883 TI - The Lymnaea respiratory system: where are we going with learning? PMID- 11729884 TI - Possible mechanisms that may determine the set point and sensitivities of the chemoreflexes. PMID- 11729885 TI - Modeling respiratory adaptations in humans. PMID- 11729886 TI - Post-hypoxic frequency decline characterized in the rat working heart brainstem preparation. PMID- 11729887 TI - Model of nonassociative learning in vagal-pontine modulation of the respiratory rhythm. PMID- 11729888 TI - NMDA receptor blockade unmasks novel gating and memory mechanisms in vagal control of respiratory rhythm. PMID- 11729889 TI - Beyond chemoreflex: plasticity, redundancy and self-organization in respiratory control: a workshop summary. PMID- 11729890 TI - Chemoreception and tonic drive in the retrotrapezoid nucleus (RTN) region of the awake rat: bicuculline and muscimol dialysis in the RTN. PMID- 11729891 TI - Modeling the ventilatory response to variations in end-tidal PCO2 in patients who have undergone bilateral carotid body resection. PMID- 11729892 TI - Cerebral blood flow sensitivities to CO2 with the steady-state method and Read's rebreathing method. PMID- 11729894 TI - Central amino acid neurotransmitters, ventilatory output and metabolism during acute hypoxia in anesthetized rats. PMID- 11729893 TI - Modeling the interaction between perfusion pressure and CO2 on cerebral blood flow. PMID- 11729895 TI - Gene regulation during intermittent hypoxia: evidence for the involvement of reactive oxygen species. PMID- 11729896 TI - Remembrance of "Oxford" conferences past. PMID- 11729897 TI - Response surface modeling of drug interactions on cardiorespiratory control. PMID- 11729898 TI - A meta-analysis of the effect of low-dose volatile anaesthetics on the ventilatory response to hypoxia: analysis for sources of heterogeneity. PMID- 11729899 TI - Simulation of the respiratory control system. PMID- 11729900 TI - Long-haul flights may induce respiratory changes similar to ventilatory acclimatisation to altitude. PMID- 11729901 TI - Sustained moderate hyperoxia augments the acute hypoxic response in awake goats. PMID- 11729902 TI - Chronic intermittent hypoxia enhances carotid body chemoreceptor response to low oxygen. PMID- 11729903 TI - Respiratory effects of breathing high oxygen during incremental exercise in humans. PMID- 11729904 TI - Intra-breath profiles of alveolar gas exchange and the initial limb-to-lung transit delay during exercise in humans: relevance for modeling the exercise hyperpnea. PMID- 11729905 TI - Skeletal muscle chemoreflex in exercise ventilatory control. PMID- 11729906 TI - Distention of venous structures in muscles as a controller of respiration. PMID- 11729907 TI - Ventilatory and circulatory responses at the onset of rapid changes in posture. PMID- 11729908 TI - Cardiorespiratory responses to walking and running at an incremental speed of treadmill. PMID- 11729909 TI - The breath sound check and exercise at or about the ventilatory threshold. PMID- 11729910 TI - Improved O2 transport and utilization capacity following intermittent hypobaric hypoxia in rats. PMID- 11729911 TI - Intercept shift in the breathlessness/ventilation relationship in response to progressive increase in workload: change in exercise breathlessness response. PMID- 11729912 TI - Ventilatory vs. dyspneic responses to augmenting and subsequently declining CO2 stimulation in humans. PMID- 11729914 TI - Hypoxic-hypercapnic interaction on ventilatory response and respiratory sensation in females: profile during menstrual cycle. PMID- 11729913 TI - Neurotransmitter release from the rabbit carotid body: differential effects of hypoxia on substance P and acetylcholine release. PMID- 11729915 TI - Opioid effect on breathing frequency and thermogenesis in mice lacking exon 2 of the mu-opioid receptor gene. PMID- 11729916 TI - Modulation of breathing using imperceptible unloading. AB - We investigated the role of V(T) and V(T)/T(I) modulation of breathing in awake human subjects. We applied a PRBS of volume (incrementing ramp) or flow (decrementing wave) assist at levels below the perceptual threshold in order to stimulate respiratory feedback. We modeled the PRBS data with linear difference equations to obtain impulse-response profiles of V(T), V(T)/T(I), T(I) and factorial(P(MUS)). We limited cortical responses to our stimuli by applying sub threshold levels of assist, and we limited humoral effects (O2 and CO2) by augmenting mechanical respiratory output intermittently and by small amounts. We found that flow or volume assist elicited similar significant increases in V(T) and V(T)/T(I). During flow assist there was a significant decrease in factorial(P(MUS)) and T(I) was reduced, albeit not significantly; however, volume assist did not modify T(I) or factorial(P(MUS)). The earlier onset of flow assist, relative to volume assist, may explain the difference between the responses. We conclude that vagally mediated inspiratory flow receptors in the chest wall or lungs may modulate breathing on a breath by breath basis when small, imperceptible increases in airflow occur early during inspiration. Furthermore, lung volume feedback during imperceptible unloading (occurring at the end of inspiration) was less effective. Finally, pseudorandom unloading with imperceptible stimuli provides a useful tool to study reflex regulation of ventilation in awake subjects without confounding cortical influences. PMID- 11729917 TI - General model for patient-ventilator interactions. PMID- 11729918 TI - Phase relations between rhythmical forearm movements and breathing in cerebellar impaired patients. PMID- 11729920 TI - The control of breathing at rest. PMID- 11729919 TI - Computational modeling of integration of voluntary/behavioral and automatic mechanisms for breathing control. PMID- 11729921 TI - Central and peripheral chemoreflex characteristics: panic disorder patients vs. healthy volunteers. PMID- 11729922 TI - A physiological and psychological model of idiopathic hyperventilation. PMID- 11729923 TI - Behavioral state control and airway instability. PMID- 11729924 TI - Muscarinic receptors influence catecholamine release from the cat carotid body during hypoxia. PMID- 11729925 TI - Neural drives and breathing stability. PMID- 11729926 TI - Variability of CO2-sensitivity during sleep. PMID- 11729927 TI - Dynamics of heart rate control in obstructive sleep apnea: a minimal model. PMID- 11729928 TI - Computational model of the effects of breathing on cardiac function. PMID- 11729929 TI - Inspiratory timing differences and regulation of Gria2 gene variation: a candidate gene hypothesis. PMID- 11729931 TI - Comparison between the respiratory inductance plethysmography signal derivative and the airflow signal. PMID- 11729930 TI - Explained and unexplained variability of CO2-sensitivity in humans. PMID- 11729932 TI - Pharmacological and immunochemical evidence of the dopamine D3 receptor in the goat carotid body. PMID- 11729933 TI - Respiratory instability and variability: why is respiration such a mess? A workshop summary. PMID- 11729934 TI - The excitatory effect of nitric oxide on carotid body chemoreception is blocked by oligomycin. PMID- 11729935 TI - CO2/HCO3- modulates K+ and Ca2+ currents in glomus cells of the carotid body. PMID- 11729936 TI - Inhibition or lack of NAD(P)H oxidase subunits do not alter the normal cytosolic calcium, sensory and respiratory response of chemoreceptors. PMID- 11729937 TI - Peripheral and central chemosensitivity: multiple mechanisms, multiple sites? A workshop summary. PMID- 11729938 TI - Role of brainstem respiratory neuron types in phase-switching produced by afferent vagal stimulation. PMID- 11729940 TI - Workshop on modeling in the 21st century: an executive summary. PMID- 11729939 TI - Modulation of the central respiratory effects of 5-HT by vagal afferents in newborn rat. PMID- 11729942 TI - Response to repeated CO2 in individuals with elevated anxiety sensitivity: replication with 20% CO2. AB - The present report replicates and extends previous work examining response patterns to repeated presentation of CO2. In previous studies, two distinct response patterns to repeated presentation of 35% CO2 were noted, representing habituation and nonhabituation of anxiety. In this report, 21 participants who had never experienced a panic attack but who reported high levels of anxiety sensitivity were presented with 12 trials of 20% CO2, followed by a trial involving inhalation of room air (to examine dishabituation) and two more trials of 20% CO2. Results indicated that 67% of the sample reported habituation of anxiety. Reductions in anxiety across inhalations were paralleled by changes in tidal volume, perceived panic symptom severity, and feelings of panic. Notable dishabituation was observed in the nonhabituation sample. Results are discussed in the light of basic learning processes underlying the treatment of Panic Disorder (PD). PMID- 11729941 TI - Activation of medullary post-inspiratory related neurons during clonidine-induced central apnea in anesthetized goats. PMID- 11729943 TI - Computer-aided vicarious exposure versus live graded exposure for spider phobia in children. AB - The efficacy of computer-aided vicarious exposure (CAVE) for the treatment of spider phobia in children was evaluated in a single blind, randomised, controlled trial. Twenty-eight participants, aged 10-17 years, received three 45-min sessions of either Live graded exposure (LGE), CAVE or were assigned to a Waitlist. Phobic symptomatology was measured at pre- and post-treatment, and at one month follow-up on a range of behavioural and subjective assessments. The results showed the superiority of the LGE treatment over the CAVE and Waitlist conditions. Effect sizes support CAVE treatment as being superior to the Waitlist and resulting in reductions of phobic symptomatology. PMID- 11729944 TI - The effect of mood and arousal on UCS expectancy biases. AB - This paper reports two studies investigating the role of mood and arousal on the development of UCS expectancy biases to fear-relevant and fear-irrelevant stimuli. Experiment 1 found that, compared with a neutral control condition, individuals in both experimentally-induced positive and negative moods significantly overestimated the possibility of aversive outcomes following all types of stimuli in a hypothetical thought' conditioning experiment. Experiment 2 found that this UCS expectancy bias produced by both positive and negative mood could not be explained by the effect that these mood states might have on arousal independently of the valency of the mood. The findings appear to be explained best by emotional response categorisation theory (Psychol. Rev. 106 (1999) 337 361), which predicts that individuals would be more likely to view emotionally valenced stimuli as similar when in an emotional state. It is argued that, while negative mood may be a genuine vulnerability factor for the development of anxiety-relevant threat-outcome expectancies, the effects found with positive mood in the current studies may have less ecological relevance to real-world aetiologies. PMID- 11729945 TI - GFP-centrin as a marker for centriole dynamics in the human breast cancer cell line MCF-7. AB - Centrosome duplication plays an important role in genomic stability through bipolar spindle formation and equal chromosome segregation during mitosis. Defects in centrosome duplication and centrosome amplification correlate with aggressive tumors and aneuploidy. Cyclin-dependent cell cycle regulators play a key role in signaling centrosome duplication and the tumor suppressor genes p53, BRCA1 and BRCA2 are suspected to function at mitotic checkpoints that monitor centrosome duplication. The relationship between loss of hormone dependence in breast cancer, and signaling of centrosome duplication in tumor progression is not known. We have developed a MCF-7 cell line expressing GFP-centrin that allows direct visualization of centriole duplication during the cell cycle in living cells. GFP-centrin is expressed and selectively incorporated into the structure of both centrioles making them clearly visible in living cells. Our studies demonstrate three important aspects of recombinant GFP-centrin incorporation into centrioles. 1) GFP-centrin transfected cells grow normally in culture and show no adverse effect associated with GFP-centrin expression; 2) newly duplicated centrioles incorporate centrin during their genesis; and 3) GFP-centrin incorporation into centrioles does not grossly affect cell cycle progression, or centrosome function. PMID- 11729946 TI - The role of androgens in mammary carcinogenesis. AB - Despite extensive research, the precise mechanism of mammary carcinogenesis is unknown. We have developed an animal model in which a high incidence of mammary cancer can be induced within a period of several months using a combination of testosterone (T) and 17beta-estradiol (E2) without the addition of carcinogens. The induced mammary tumours mimic closely the human breast cancer in terms of histopathology. Our results showed that the two sex hormones work synergistically to induce a higher incidence of mammary cancer than either hormone treatment alone. The dosage of T affects only the latency period of mammary cancer but not the final incidence. The results further showed that treatment of T, either alone or in combination with E2, there was overexpression of the androgen receptor (AR) in alveolar or ductal epithelial cells but not in stromal cells. Together with overexpression of AR in epithelial cells, there was an increase in perialveolar and interlobular connective tissue as well as a decrease in surrounding adipose tissue, despite the absence of AR in stromal cells. There was also an increase in proliferation rate of fibroblast-like cells in stroma. These changes were blocked by implantation of flutamide, an antiandrogen, indicating that androgens play a crucial role in the process. These findings highlight that the effect of androgens on the stroma, may be through a paracrine action of epithelial cells. The changes in the stroma may, in turn, promote mammary carcinogeneis in a reciprocal manner. PMID- 11729947 TI - Alkaline phosphatase and tartrate-resistant acid phosphatase in osteoblasts of normal and pathologic bone. AB - A review of histochemical and immunohistochemical studies on alkaline phosphatase (AP) and tartrate-resistant acid phosphatase (TRAP) in osteoblasts leads to the following conclusions: 1) AP is a typical (non-specific) marker of osteoblasts, and TRAP is a typical (non-specific) marker of osteoclasts; 2) both enzymes may be used to identify the precursors of these cells and are found in young osteocytes; 3) both are released into the extracellular space; 4) both are expressed by metaphyseal osteoblast-like cells whose ultrastructural characteristics are similar to those of post-osteoclastic cells of the basic multicellular unit (BMU) and are also exhibited by osteoblasts and macrophages; 5) the increased numbers of these cells in hypocalcemic animals suggests that the local calcium ion concentration may trigger the transition of the reversal into the formation phase of the BMU. PMID- 11729948 TI - Three dimensional ultrastructure of transverse (Harris) lines in the long bone. AB - To investigate the occurrence of transverse lines in long bones of adults, we studied 27 cadavers (14 males and 13 females). After confirming the presence of a transverse line, a cross-sectional sample was examined macroscopically, and by soft X-ray and scanning electron microscopy. The following results were obtained: 1. According to roentgenograms, transverse (Harris') lines were observed in 40.7% of the distal half of the femur and in 29.6% of the proximal half of the tibia. 2. Macroscopic examination of the bone cross-sections at the level of the transverse line showed various membranous structures. 3. In scanning electron micrographs, no marked difference in structure was observed between the transverse line trabeculae and the compact bone. PMID- 11729949 TI - Ultrastructural analysis of bone calcification by using energy-filtering transmission electron microscopy. AB - In order to elucidate the mechanisms of bone calcification, embryonic rat calvariae treated with chemical or cryo-fixation were observed using transmission electron microscopy by three techniques: fine structures, various cvtochemical localizations including nonspecific proteoglycan, decorin, chondroitin 4-sulfate, hyaluronan, alkaline phosphatase (ALP), and osteonectin, as well as the elemental mapping of calcium and phosphorus by energy-filtering electron microscopy. In the calvariae, the calcification sequence ran as follows crystallization within matrix vesicles, formation of calcified nodules, collagen calcification, and finally the establishment of an expansive calcified matrix. The osteoid contained an abundance of mesh-like fibers of proteoglycans, including decorin, chondroitin 4-sulfate, and hyaluronan, around collagen fibrils approximately 50 nm in diameter. Calcium tended to localize at the proteoglycan sites, while phosphorus was often mapped to the collagen fibril-structures in the osteoid. Calcium/phosphorus co-localization was found in and around the calcified nodules, where ALP and small sized proteoglycans were observed. During this stage, native proteoglycans surrounding the collagen fibrils disappeared, with the collagen fibrils fusing laterally, and attaining a diameter of more than 400nm. The calcified nodules expanded to occupy the entire space made available by the collagen fibril-fusion, following osteonectin accumulation in the calcified nodule/collagen fibril border. In conclusion, crystals present within the matrix vesicles became calcified nodules, in a process induced by the co-localization of calcium and phosphorus. ALP and proteoglycans may participate in the calcium/phosphorus co-localization. Decreases in the native proteoglycans, and the lateral fusion of collagen fibrils are thought to be involved in the expansion of calcified areas, followed by osteonectin-mediated collagen calcification. PMID- 11729950 TI - Entheses--the bony attachments of tendons and ligaments. AB - Most tendons and ligaments have fibrocartilaginous entheses where there are 4 zones of tissue at their bony attachments--dense fibrous connective tissue, uncalcified fibrocartilage, calcified fibrocartilage and bone. Such entheses leave smooth, circumscribed markings on dried bones. The uncalcified fibrocartilage dissipates the bending of collagen fibres away from the bone, ensures that a stretched tendon or ligament does not narrow too close to the bone and acts as a mini growth plate. The zone is thickest at entheses where a great deal of bending of the tendon/ligament accompanies joint movement. The calcified fibrocartilage anchors the tendon/ligament to the bone and enables it to withstand shear. Enthesis fibrocartilage may be accompanied by sesamoid and periosteal fibrocartilages that similarly protect the enthesis from wear and tear and dissipate stress. Nevertheless, each fibrocartilage can show distinctive pathological changes. A wide variety of ECM molecules has been reported in enthesis fibrocartilage, but it is best characterised by its content of type II collagen and aggrecan which account for its compression-tolerance properties. PMID- 11729951 TI - Altered balance of proteinase inhibitors in atrophic muscle after denervation. AB - Skeletal muscle denervation leads to an increase of proteolytic activity, which is also favoured by reduced levels of alpha1 antichymotrypsin and nexin II, two serine-proteinase inhibitors normally acting at the neuromuscular junction. In the present experiments we extended our investigation to other muscular proteinase inhibitors after denervation. In all muscles examined (soleus, plantaris, extensor digitorum longus) specific immunoreactivity for alpha2macroglobulin (alpha2M) and alpha1proteinase inhibitor (alpha-1 antitrypsin, ATI) was distributed in peri-endomysial structures as well as in small patches inside the fibres. By contrast, inter-alpha-trypsin inhibitor (ITI) was mainly localized in the extracellular matrix. These localization patterns did not change substantially in 15-days denervated muscles. Dot-blot analysis revealed a small decrease (about 15%) of alpha2M in 15-days denervated muscles, while ATI and ITI specific activities were substantially unchanged. RT-PCR allowed us to detect the above protease inhibitor mRNAs in normal muscle homogenates. Denervation atrophy induced by section of the sciatic nerve resulted in a remarkable reduction of (2macroglobulin mRNA (60%) and ITI (30%), but not ATI, as measured by computer-assisted semiquantitative densitometry of electrophoresed RT-PCR bands. The marked decrease of alpha2M we have detected in denervated muscle may be responsible, at least in part, for the proteolytic increase which is known to occur in skeletal muscle during denervation atrophy. PMID- 11729952 TI - Three-dimensional organization of mammalian skeletal muscle membrane systems. AB - The three-dimensional arrangement of mitochondria and sarcoplasmic reticulum membrane systems of vertebrate extrafusal and intrafusal muscle fiber types was studied by scanning electron microscopy (SEM) in the specimens prepared by the A ODO method. SEM observations on specimens treated by HCl-digestion method revealed distinct differences in motor endplates in the different muscle fiber types. PMID- 11729953 TI - The effect of anabolic steroid on the distribution of muscle fiber in rat hind limb. AB - This study evaluates the effect of anabolic steroid hormone (nandrolone decanoate, 19-Nortestosterone; ND) on histomorphometry and distribution of muscle fiber in the rat tibialis anterior (TA) muscle compared with the control rats. Three male Wistar rats aged of 5 weeks were daily received subcutaneous injection of 0.3 cc vehicle oil included 1mg ND for 6 weeks. Two control (C) rats were done only vehicle oil by the same manner as the experimental (E) group. The TA muscle was stained for myofibrillar adenosine triphosphatease with preincubation at pH 4.35, after cryosectioning at the thickness of 10 microm. Muscle fibers were classified into type I, IIA, IIX, and IIB. The E group showed significantly smaller than the C group in body weight. In histomorphometry, the cross-sectional area of type IIB fibers in the E group showed narrow range, and shifted smaller than those in the C groups. The distribution of type I fibers was restricted to the deep region in the E group. The anabolic steroid facilitated toward the older rat type on distribution and proportion of muscular tissue. PMID- 11729954 TI - The epicardium as a source of mesenchyme for the developing heart. AB - The primitive epicardium of the vertebrate embryo has traditionally been regarded as a rather passive mesothelium, lining the embryonic myocardium and forming the adult visceral pericardium. However, in recent years, there is an increasing evidence that the primitive epicardium is a highly dynamic element which supplies cells to the developing heart through a process of epithelial-mesenchymal transition. This process seems to be more active at the atrioventricular canal and outflow tract, i.e. the cardiac segments where the endothelium transforms into mesenchyme. In this paper we review the current evidence which supports such epicardial-mesenchymal transition, namely: 1) morphological features, 2) colocalization of cytokeratin and vimentin in the epicardial and subepicardial mesenchymal cells, 3) presence of common antigens in the transforming epicardium and endocardial cushions (fibrillin-2/JB3, ES/130, Ets-1). Recendy, we have immunolocated the transcription factor Slug in the developing avian heart. Slug is a zinc-finger protein involved in the formation of the neural crest, a developmental event which implies an epithelial-mesenchymal transition. All cells of the primitive epicardium are Slug+ from their differentiation until the stage HH24. However, only a fraction of the endothelial cells from the endocardial cushions are Slug+. We speculate that the expression of Slug marks competence of the epicardial cells to transform into mesenchyme, although this transformation is only achieved where an inducing signal is produced. Regarding the developmental fate of the epicardial-derived cell population, there is strong evidence of its differentiation in fibroblasts and vascular smooth muscle cells, although a contribution to the coronary endothelium cannot be discarded. PMID- 11729955 TI - A KOH-collagenase digestion method for scanning electron microscopic studies of vascular smooth muscle fibers in the human heart. AB - The three-dimensional cytoarchitecture of smooth muscle fibers of human cardiac arterial vessels was studied by scanning electron microscopy after removal of extracellular connective tissue matrices with a KOH-collagenase digestion method. Arterioles with an outer diameter of 30-100 microm had a well-developed compact media consisting commonly of circularly oriented smooth muscle fibers. There were also arterioles with oblique or longitudinal muscle fibers in some places. In terminal arterioles with an outer diameter of less than 30 microm, muscle fibers became branched and commonly encircled the endothelial tube. There were some terminal arterioles covered with obliquely arranged smooth muscle fibers. The presence of the regional difference in arrangement of vascular smooth muscle fibers indicates the heterogeneity of blood flows in the level of cardiac microcirculation. PMID- 11729956 TI - Developmental morphology of blood and lymphatic capillary networks in mammalian hearts, with special reference to three-dimensional architecture. AB - The development of blood and lymphatic capillaries in the cardiac ventricles was studied using the rat and monkey hearts from later gestation to adulthood. For scanning electron microscopy, the tissue was treated with ultrasonication followed by HCl in order to remove epicardial mesothelial cell, subepicardial connective tissue and basement membrane. At the end of fetal life and neonate, the blood capillaries ran in parallel between the bundles consisting of several cardiac myocytes with small size. Endothelial extensions were found frequently. Numerous pericytes with short processes were observed on the capillary wall. Blood capillaries in the pubertal myocardium ran between cardiac myocytes to form a dense network. Processes of pericytes became elongated along the capillaries. In the adult, the capillaries showed occasionally Y-shaped branching and H-shaped anastomoses. The lymphatic capillaries were identified by large, variable sizes and absence of pericytes. In early postnatal rats, they appeared in the subepicardial region. In young and adult animals, the lymphatic capillary networks with relatively dense reticular meshes surrounded the bundles of several myocytes. It was noted that numerous nerve fibers were in close to the lymphatic capillary wall. PMID- 11729957 TI - Towards a dynamical approach for the simulation of large scale, cancer correlated chromatin structures. AB - To understand the influence of geometrical constraints in the spatial distribution of cancer correlated "Double Minute chromosomes (DMs)" in human cell nuclei, we applied computer simulations of the nuclear 3D structure in combination with a voxel based segmentation algorithm. With this approach we determined the overlap volumes and intensities of the DMs with the chromatin free space in the simulated nucleus. For this purpose, beginning from a start configuration, simulated linear chromosome chains together with the DMs were relaxed according to the Monte Carlo process. The simulations predict a preferential positioning of DMs within the "peripheral" "Inter Chromatin Domain (ICD)" space. PMID- 11729958 TI - Quantitative microvascular corrosion casting by 2D- and 3D-morphometry. AB - As a system of tubes (blood vessels) the cardiovascular system changes actively and passively diameters to adapt its transport capacities for respiratory gases, nutrients, heat, metabolites and waste products to and off the body's organs, tissues and cells. In most healthy organs blood vessels form a hierarchically arranged three-dimensional network with the geometry defined by vessel diameters, interbranching distances (defining branching frequencies and number of branching sites, i.e. nodes), intervascular distances, and branching angles. In the present study 2D- and 3D-morphometry is applied to quantify these parameters and their changes as they occur in resin casts during metamorphosis of the tadpole lung (2D morphometry) and filter apparatus vasculature (3D-morphometry). It is shown that 2D-morphometry should be limited to the analysis of high powered images of flat two-dimensional vascular networks (example: tadpole lung alveolar vascular bed) to prevent underestimation of parameters. In contrast, 3D-morphometry can be applied over a wide range of magnifications whereby accuracy of measurements increases with the portion the structure to be measured occupies within the field of view. Together with a careful control of precasting conditions (application of vasoactive drugs, anaesthetics), casting conditions (pressure during rinsing and casting, amount of final shrinkage of casting media), and postcasting conditions (thermal burdening during maceration, sputtering, evaporation, and SEM inspection; thickness of conductive metal layers) 3D-morphometry enables to gain reliable data from resin casts of highly complex real vascular networks in healthy and diseased organs in the developing, juvenile, adult and aged state, as well as in different physiological states. PMID- 11729959 TI - Vascular architecture of normal human urinary bladder and its remodeling in cancer, as revealed by corrosion casting. AB - The vasculature of normal human urinary bladder wall and its tumors were studied using corrosion casting and scanning electron microscopy. In the normal wall, a regular organization of highly tortuous horizontal and vertical vessels allows accommodation of the vascular system to spatial changes resulting from the filling/voiding cycle. The vasculature of tumors is a result of remodeling of preexisting wall vessels associated with gradual growth of the neoplastic tissue. The shape and arrangement of blood vessels in different regions of the tumor seem to reflect a variable dynamics of tumor development and possible influence of various angiogenesis-promoting growth factors. PMID- 11729960 TI - Morphostructural changes of the kidney microvasculature during the prenatal period in Bubalus bubalis. AB - The microcirculation of the foetal kidney was studied in the buffalo using light (LM) and scanning electron microscopy (SEM). The primordial glomerules originated from the peripheral zone of the metanephros at the stage of 8 cm CRT. The glomerular capillaries started to differentiate at the stage of 10-15 cm CRT. They were sparse and showed a few primordial pores. In addition, they began to make contacts with primordial podocytes. At the stage of 40-60 cm CRT, the renal microcirculation showed a complex and almost completely organized morphology. PMID- 11729961 TI - Retinal vascular changes and systemic diseases: corrosion cast demonstration. AB - The purpose of the present study is to clarify the specific retinal vascular changes in rat models of single spontaneous or hereditary systemic diseases. We used Wistar Kyoto (WKy) rats 6 months of age as normal controls, 24-month-old Wistar Kyoto rats in studies of aging, 16-month-old spontaneously hypertensive rats (SHR), 18-month-old rats with inherited hypercholesterolemia (RICO) for arteriosclerosis, and 18-month-old Otsuka Long-Evans Tokushima Fatty (OLETF) rats for diabetes mellitus. Retinal vascular corrosion casts were made and observed with a scanning electron microscope. The retinal vessels were also examined with a transmission electron microscope. Specific changes in the retinal vessels were: narrowing and thin thread-like capillaries in aging; capillary tortuosity, irregularity and narrowing in hypertension; straightening in hypercholesterolemia; and loop formation and microaneurysms in diabetes mellitus. These specific changes in the retinal vessels in each systemic disease can be clearly and easily revealed in a three-dimensional fashion by corrosion casts followed by scanning electron microscopy. PMID- 11729962 TI - Corrosion cast demonstration of choroidal vasculature in normal Wistar Kyoto rat. AB - No previous attempt has been made so to demonstrate the details of the choroid vascularization in the rat. This paper describes the choroidal vascular pattern of the normal Wistar Kyoto rat using the corrosion cast method. 6-month-old normal Wistar Kyoto rats were used. Vascular casts prepared by our previously described technique were observed with a Hitachi S-2360N scanning electron microscope. Corrosion casts of the entire choroidal vasculature showed that the ophthalmic artery branched into two main ciliary arteries: the nasal and temporal arteries, which extend forward to form the iridociliary circle. Further the inferior ciliary artery arises from the temporal ciliary artery. In the posterior eye segment these arteries form four to seven branches supplying the adjacent choriocapillaris. There are different arrangements of choriocapillaris. The larger veins formed four vortex veins, one in each quadrant, draining blood from the anterior region of the choroid or iris. Venous blood from the central region, peripapillary choroid and sometimes optic nerve head is drained by smaller veins, which run more or less directly into the posterior ciliary veins. In conclusion corrosion casts and SEM can show the details of the choroidal vascular architecture. These three-dimensional (3-D) findings of choroidal vessels of the rat are very useful for the study of choroid vascular abnormalities in various pathological conditions. PMID- 11729963 TI - Phylogeny and ontogeny of the lymphatic stomata connecting the pleural and peritoneal cavities with the lymphatic system--a review. AB - This paper reviews the phylogeny and ontogeny of "lymphatic stomata" through which fluids and cells in the pleural and peritoneal cavities enter the lymphatic system. In amphibians, the pleuroperitoneal cavity is connected through numerous pores with the wide subvertebral lymphatic sinus corresponding to the thoracic duct in mammals. In reptiles, direct connections of the pleural and peritoneal cavities with the lymphatic system have been reported. In mammals, the pleural and peritoneal cavities are directly connected with lymphatics through lymphatic stomata. How do lymphatic stomata develop in mammals? In the rat, distinct lymphatics are noted in the subpleural space of the diaphragm periphery in 16 days old embryo. With age, the supleural lymphatics increase and form a polygonal network. They show a tubular appearance and possess many valves. Some of them become endowed with smooth muscle cells. In 19 days old embryos, some lymphatics appear in the subperitoneal space of the diaphragm. They extend centripetally and form many lateral projections that later elongate and connect with those from adjacent lymphatics, thus forming a lattice-like network or "lymphatic lacunae". During early postnatal days, the lymphatic lacunae project many bulges that subsequently come into contact with the pores among mesothelial cells lining the diaphragmatic peritoneum, thus forming lymphatic stomata. They increase until postnatal week 10. The lymphatic stomata in the costal pleura also develop during early postnatal days. PMID- 11729964 TI - Re-evaluation of the reticulo-endothelial system. AB - By injecting lithium carmine (Lit-car) into living animals, Ribbert (1904) and Kiyono (1914) showed that specific staining occurred in some cells in various organs. Kiyono termed those cells "histiocytes" which consisted of free amoeboid cells and cells of reticulo-endothelium. Aschoff (1924) introduced the concept 'reticulo-endothelial system (RES)' for the collection of cells having in common the property of vital staining. Van Furth (1972) proposed the term "mononuclear phagocyte system (MPS)" advocated that RES be replaced by MPS. As a consequence we presently suffer from a "reticulo-endothelial confusion", based on the delusion that cells other than macrophages are not members of the RES. The point that most clearly illustrates this is the fact that the Lit-car-laden cells in the body are named macrophages in modern textbooks of histology. To clarify the confusion, we re-examined the vital staining at light and electron microscopic levels and analyzed the scavenger cells using biochemical methods. Injected Lit car was actively endocytosed by sinusoidal endothelial cells in the liver and reticular cells in the sinus of lymph nodes. Of note, uptake of the dye was comparatively much lower in macrophages/monocytes. Our findings indicate the existence of scavenger endothelial cells and reticular cells in blood and lymphatic circulations respectively. We name these two systems "the scavenger RES" collectively. PMID- 11729965 TI - Ontogeny of the dendritic, and follicle-associated epithelial cells in the bursa of Fabricius of guinea fowl (Numida meleagris). AB - A panel of monoclonal antibodies was produced to study the mesenchymal stromal elements of the bursa of Fabricius in guinea hen. The intracellular antigens recognized by GIIF3 and NIC2 monoclonal antibodies are of 50 and 30 kD, respectively. The cells identified by these antibodies emerge in the mesenchyme around day 12 of incubation, and immigrate into the surface epithelium of the bursal folds, which precedes the follicle formation. The GIIF3 cells move up to the luminal surface of the follicle and differentiate to follicle-associated epithelial cells. The NIC2 cells remain in the medulla, produce and secrete large amount NIC2 positive substance, when the bursal function starts up. The presence of double positive (GIIF3 and NIC2) cells in the medulla around hatching, seems to indicate, that the two cells might have a common origin. The NIC2 positive product of the bursal secretory dendritic cells contributes to the microenvironment, and possibly necessary for the B cell clonal expansion and establisment of the immune repertoire in the guinea hen. PMID- 11729966 TI - Involvement of the NGF receptors (Trka and p75lngfr) in the development and maintenance of the thymus. AB - Nerve growth factor (NGF) and its main low- (p75LNGFR) and high-affinity (TrkA) receptors have been found in the vertebrate thymus, thus suggesting they are involved in the control of thymic function. However, its role in this organ is poorly known. In the present study we used combined morphological and immunohistochemical techniques to analyze the distribution of TrkA and p75LNGFR in the rat thymus, as well as the structural changes in the thymus of p75 LNGFR or TrkA deficient mice. In adult rats both TrkA and p75LNGFR were localized in a subset of thymic epithelial cells found primarily in the subcapsular and medullary thymic regions, regarded to be endodermal-derived cells. Consistently, animals with a non-functional TrkA, but not those lacking p75LNGFR, showed structural changes consisting of a decrease in the density of thymocytes, absence of cortico-medullary border, and large cysts lined of endodermal epithelium. These results strongly suggest a function of the TrkA-NGF system in thymic functions mediated by epithelial cells, as well as a role of TrkA in the development of the murine thymus. The function of p75LNGFR remains to be established. PMID- 11729967 TI - Ultrastructural morphometric analysis of thymus epithelial cells two months after pinealectomy. AB - The late effect of surgical ablation of the pineal gland on the morphometric changes of epithelial cells of rat thymus were investigated. The aim of this study was to determine a possible existence of sex related changes in the thymus epithelial cell after pinealectomy and to possibly clarify the correlation between the pineal gland and the thymus. Stereological ultrastructural parameters of the thymus cortical and medullar epithelial cells of male and female rats two months after pinealectomy are reported. A group of animals was submitted to surgery for ablation of pineal gland. The control group (shame-pinealectomized animals) underwent to the same surgical procedure but without removal of pineal gland. Animals were sacrificed 60th days after surgery. Para-sagittal thymus specimens were fixed by immersion in glutaraldehyde and processed for transmission electron microscopy. The volume and surface density (Vv, Sv) of nucleus and cytoplasm of cortical and medullar thymus epithelial cells were calculated using the Weibel's multipurpose test system and multilevel sampling technique on electron micrographs. Volume and surface density of mitochondria (Vvm, Svm), endoplasmic reticulum (Vvr, Svr), vacuole (Vvv) as well as numerical density of mitochondria (Nvm) were evaluated at different magnification levels. Our analysis confirmed a statistically significant increase in Vv of endoplasmic reticulum and vacuoles in both sex of pinealectomized rats. Sv of plasmalema, endoplasmic reticulum and mitochondrial membrane were markedly increased in thymus medullar epithelial cells of pinealectomized rats. Vv of mitochondria was significantly increased in cortical epithelial cells of pinealectomized animals. These results confirm that a correlation between pineal gland and thymus exists. However the present findings seem to support the concept of sex independent inhibitory action of pineal gland on thymus cortical and medullar epithelial cells. PMID- 11729968 TI - Hemopoiesis in the liver of adult tumor-bearing mice. AB - In the liver of adult mice bearing an Ehrlich carcinoma on the leg, progressively hypoxic and displaying reactive hepatitis but not metastatic dissemination, extramedullary hemopoiesis was detected. Electron microscopy revealed mainly erythropoietic islands and scattered megakaryocytes in maturation stages up to the platelet-releasing phase. Erythropoietic cells expressed an embryonic-type of hemoglobin, which is more adequate to oxygenate hypoxic environments than the adult type. They were positive for the peroxidase reaction due to the presence of hemoglobin and could furthermore be visualized by the blue-excited red autofluorescence of protoporphyrin IX. Extramedullary hemopoiesis, one of the various examples of reactivation of fetal features in the liver associated with carcinogenesis, is supposed to be compensatory for the loss of blood cells induced by the tumor. Reviewing this process has the purpose of raising the question whether the fetal features are better adapted than adult ones to the metabolic and physiological characteristics of a tumor-influenced organism. PMID- 11729969 TI - Atomic force microscopy and its related techniques in biomedicine. AB - The atomic force microscope (AFM), invented in 1986, is a new instrument which provides three-dimensional surface images of samples by scanning a sharp probing tip over the sample surface. Unlike electron microscopes (EM), the AFM has the advantage to obtain high-resolution pictures not only in a vacuous but also in a non-vacuous (i.e., air or liquid) environment. This paper reviews our recent studies on the AFM application to the biomedical fields. AFM is useful for observing biological structures such as DNA, collagen molecules, collagen fibrils and chromosomes. AFM images of living cultured cells in liquid can be used for investigating the movement of cellular processes in relation to subcellular cytoskeletal elements. Recently, numerous AFM-related microscopes, or scanning probe microscopes (SPM), have been invented in parallel with the development of the AFM itself. These microscopes allow the simultaneous collection of topographical and other (e.g., viscoelastic, near-field optical) images of samples in the same portions. Thus, the combination of AFM and the other SPM has great potential for providing valuable new findings on structure and function of cells and tissues. PMID- 11729970 TI - Stereo-structural study of the lingual papillae and their connective tissue cores in relation to ageing changes in the human tongue. AB - This study describes the stereo structure of human lingual papillae along lifespan with particular emphasis to ageing. The following results were obtained. Numerous slender protrusions extend from the basal column of each filiform papilla of young human tongues and decrease in number with age. Many cases having an entirely flat area on the surface of the tongue without papillae were found in old age (Loss of protrusions and flattened epithelium). The number, thickness and length of each secondary protrusion of connective tissue core (CTC) of the filiform papilla decreases roughly proportionally with age (Ramification at the end of protrusions, and fused patterns of CTC of the filiform papilla). The CTC of fungiform papillae which have a corolliform structure, become thin with age. Numerous small rod-shaped secondary CTC are distributed under the epithelium of the vallate papillae in the young subjects and decrease in number and become irregularly distributed in older age. The fact that there is a great deal of individual variation of human lingual papillae regardless of age must be emphasized. PMID- 11729971 TI - Nuclear localization and signalling activity of inositol lipids. AB - It has been recently shown that also the nucleus is a site for both synthesis and hydrolysis of the phosphorylated forms of phosphatidylinositol. Among the enzymes of the cycle we and others have demonstrated that phospholipase C specific for inositol lipids (PLC) is one of the main steps of the inositol lipid cycle. The PLC family and namely the type raised a noticeable interest since, because of their common COOH-terminus, it has been shown a nuclear localisation in addition to that at the plasma membrane. The autonomous intranuclear inositide cycle is endowed, besides the PLC, with conventional lipid kinases and phosphatidylinositol 3 kinase (PI 3-kinase) which plays an important role in granulocytic differentiation. The combination of morphology and molecular biology gave us the opportunity to localise the sites inside the nucleus where the cycle takes place and to understand the physiological significance of such a subcellular compartmentalisation both during cell growth and differentiation. PMID- 11729972 TI - Structural organization of the enteric nervous system in the cattle esophagus revealed by wholemount immunohistochemistry. AB - In ruminants the motility patterns of the esophageal tube are characterized by physiological regurgitations including both anterograde and retrograde peristaltic movements. These complex motor functions require an elaborated enteric nervous system (ENS) for the generation of the underlying intrinsic reflex circuits. The structural organization of the esophageal ENS was studied in fetuses of cattle (n=6) by means of wholemount preparations obtained from different segments of the esophagus. Demonstration of nerve cells, ganglia and nerve fibers strands (NFS) was achieved by immunohistochemistry using the general neuronal marker protein gene product (PGP) 9.5. The myenteric plexus represented the most prominent nerve network composed of differently shaped ganglia and interconnecting NFS. Frequenitly the myenteric ganglia were arranged in two separate layers interweaving with the adjacent muscle coat. From the cervical towards the thoracic segment of the esophagus the density and size of myenteric ganglia increased and the NFS exhibited thicker diameters. The submucosal and mucosal plexus consisted of NFS ramifying throughout the tela submucosa and the lamina propria mucosae. The networks showed no evidence of ganglia nor single nerve cells. The findings illustrate that intrinsic esophageal nerve cells are confined to the myenteric plexus. Since the esophageal tube has no secretory functions, secreto-motor neurons are not required in the submucosal and mucosal plexus layers. The structural organization of the intramural nerve networks--in particular the specific arrangement of the myenteric plexus--reflects the substantial contribution of the esophageal ENS to the coordination and mediation of esophageal motility in ruminants. PMID- 11729973 TI - The tubulovesicular system of gastric parietal cells is connected to the intracellular canaliculus, rough endoplasmic reticulum and Golgi complex. A study by high resolution scanning electron microscopy. AB - The three-dimensional configuration of the tubulovesicular system of resting rat gastric parietal cells was determined by ultra-hihg-resolution scanning electron microscopy (SEM). Rat gastric mucosae were fixed in 1.0% glutaraldehyde and 0.5% formaldehyde in cacodylate buffer, (334 milliosmoles/kgH2O (mOsm)). To render cytoplasmic membranes visible by SEM, fixed mucosae were frozen and fractured, and the cytoplasm of fractured parietal cells was macerated by the aldehyde osmium-DMSO-osmium procedure. With much of the cell matrix and filaments removed, SEM revealed numerous 30-60 nm tubukes, which formed a meshwork and also small cisternae about 100 nm. Some cisternae had a small, central 10 nm fenestration. The cytoplasmic surface of the tubules was smooth surfaced but some cisternal areas had attached polyribosomes. Isolated vesicles or tubules were not found in adequately macerated parietal cells. The tubulocisternal network was also connected to the Golgi apparatus. In favorable sites connections of the tubular membranes to the canaliculi were clearly visible. Continuity between these two membrane compartments suggests the probability of rapid membrane transposition when cells are stimulated to secrete acid. PMID- 11729974 TI - Usefulness of endoscopic small intestinal biopsies in children with coeliac disease. AB - Small intestinal biopsy is the most important diagnostic method in the routine evaluation of children with chronic diarrhoea and malabsorption. At present morphological alterations are considered essential in the diagnosis of coeliac disease (CD) and the presence of a normal small bowel biopsy specimen, observed in patients eating a diet containing gluten, rules out the diagnosis of CD. The small intestinal biopsy can be carried out either by blind suction capsule or by endoscopic forceps. In everyday clinical practice endoscopic duodenal biopsies, if taken and handled suitably, are accepted as equivalent to capsule biopsies from the proximal jejunum. In the study we reported some patients in whom has been possible to demonstrate the presence of total villous atrophy in one biopsy, while other duodenal samples taken in different duodenal portions were normal or showed mild lymphocytes and plasmacells infiltrations of the lamina propria. In patients with this type of biopsy pathology, wherein flat mucosa has been found even close to normal mucosa, the possible explanation is mucosal patchiness. The occurrence of patchly distributed intestinal atrophy in children suffering of CD raises the question of the validity of using the peroral capsule, widely believed to be the best standard for the diagnosis of CD. In our opinion, small intestinal biopsies obtained via endoscopy are more reliable than the peroral capsule biopsies in order to identify patchy mucosal atrophy and could be very useful for a correct diagnosis in CD patients. PMID- 11729975 TI - Fractal geometry: a useful tool for quantifying irregular lesions in human liver biopsy specimens. AB - Irregularity and complexity are the main features of every biological system, including human tissues, cells and sub-cellular components. These two properties of the organized biological matter cannot be quantified by means of the classical Euclidean geometry, which is able to measure regular object, practically unknown in Nature. The aims of our paper were a) to underline the importance of the shape of a biological structure, b) to investigate the fractal geometry for quantifying the liver histo-pathological structures, and c) to explain the significance of several terms used in the fractal analysis of complex biological systems. PMID- 11729976 TI - Apoptosis induction in vivo and fate of apoptotic material in the colon of the guinea pig. AB - Short-chain fatty acids (SCFA) in particular butyrate are regarded as an energy source acting in beneficial, protective manner on the colonic mucosa. Previous investigations showed that the colonic mucosa bathed in Ussing chamber with a solution lacking butyrate induced massive apoptosis of epithelial cells. The apoptotic material (bodies and cells) was shed at the mucosa surface. In the present study we aimed to investigate the effects caused in vivo on the colonic mucosa by the absence of butyrate. For this purpose the colon of guinea pigs was perfused in situ with solutions either containing or lacking butyrate. The results show that within 2h of perfusion without butyrate a large amount of epithelial cells underwent apoptosis as in the in vitro experiments. However, apoptotic material instead to be extruded at the epithelial surface accumulates into the intercellular spaces from which it becomes removed by an unusual high number of macrophages. These, engorged with phagocytozed material, lie assembled in a layer below the epithelium. Similar alterations have not been observed after perfusion in the presence of butyrate. The results suggest that this SCFA may protect the colonic mucosa in that it prevents apoptosis. The alterations occurring during 2h of its absence allow to assume that a protracted butyrate deprivation may lead to a breakdown of the integrity of the mucosa thus influencing differently the activity of the macrophages. PMID- 11729977 TI - Epithelial cell turnover--extracellular matrix relationship in the small intestine of human adults. AB - In the human small intestine, proliferation, migration, differentiation and death of epithelial cells take place in separated compartments along the crypt-villus axis. It has been shown in different cell systems that these basic biological activities are influenced by extracellular matrix proteins. To investigate possible relationships in the epithelium of the human adult small intestine we examined immunohistochemically the distribution of type IV collagen, laminin, fibronectin and tenascin, and compared the sites of their expression with the various cell activities. Epithelial cell proliferation and cell death have been detected by an antibody against Ki-67 and the TUNEL-assay, respectively. The results show that Ki-67 staining is restricted to the crypts and TUNEL-positive cells are only present in the upper villus region. Type IV collagen is uniformly present in the epithelial basement membrane along the crypt-villus axis providing a scaffold for other components of the extracellular matrix. Laminin appears to be associated with epithelial cell differentiation, since it is strongly expressed in the villus basement membrane but only weakly underneath the crypt epithelium. Although fibronectin displays a staining pattern similar to that of laminin, it might rather be responsible for cell adhesion. Strong indications have been found that tenascin could be related to epithelial cell death since it was particularly expressed at the villus tip, where the cells undergo apoptosis. PMID- 11729978 TI - Morphological features and modulation of the intrahepatic biliary epithelium. AB - Much has been learned in the past few years concerning the morphology and function of the intrahepatic biliary epithelium. Immunohistochemistry, together with ultrastructural studies has allowed a better identification or the smallest branches of the biliary tree and of subcellular components (cytoskeleton, specializations of the cell membrane, specific receptors). Modulation of the biliary epithelium in response to physiological or pathological stimuli has renewed the interest concerning the existence of facultative stem cells in the liver. More information is needed however concerning the mechanisms of cell loss in vanishing bile duct syndromes. PMID- 11729979 TI - Immunohistochemical features of bile duct epithelial cells in normal and experimental liver conditions. AB - It is well known that estrogen (E) modulates the processes of liver growth and regeneration. However, while estrogen receptors (Er) have been detected in hepatocytes, little is known on the occurrence of Er in cholangiocytes and the role of E on the physiopathology of the biliary epithelium. The purpose of this study was to investigate the occurrence of Er and their alpha or beta subtypes in cholangiocytes of normal and Bile Duct Ligated (BDL) rats and to evaluate the role and mechanisms of E in the modulation of cholangiocyte proliferation. In this study normal and BDL rats (utilized as experimental model of cholestasis) were used. Er alpha and beta subtypes, CK-19, PCNA and Fas were analysed by immunohistochemistry. The antiestrogens tamoxifen or ICI 182,780 were administered in the BDL group and the effects on cholangiocyte proliferation (bile duct mass) and apoptotic phenomenon (Tunel and Fas expression) were evaluated. Our results demonstrated that cholangiocytes express both Er-alpha and Er-beta subtypes, while hepatocytes only express Er-alpha. The increased percentage of cholangiocytes during BDL-induced proliferation was correlated with Er and PCNA expression and with enlarged Bile Duct Mass (BDM). Treatment of BDL rats with antiestrogens induced: i) inhibition of cholangiocyte proliferadon as indicated by the decreased BDM and PCNA expression; ii) over-expression of Fas antigen in cholangiocytes and induction of apoptosis (TUNEL) and iii) inhibition of cholangiocyte secretory activities. In condusion, our findings demonstrate that cholangiocytes express Er which are up-regulated during cholangiocyte proliferation. Inhibition of Er with antiestrogens blocks cholangiocyte proliferation and triggers apoptosis of Fas+ cholangiocytes suggesting a crucial role of estrogens in modulating cholangiocyte proliferation during bile duct obstruction. PMID- 11729980 TI - Repulsive charge mechanism serve to maintain lumens and cavities. An histochemical study of rat serosa and kidney. AB - The free surface of the rat peritoneum was covered with a rich negative-charged substance which is distinctly stained with cationic colloidal iron (pH 1.5). Neuraminidase digestion erased this iron stain. Treatment with Limax flavus agglutinin (LFA), which has specific affinity to sialic acid, interferred with iron staining on the serosal surface. Transmission electron microscopy of the peritoneal samples stained with colloidal iron showed that colloidal particles were deposited on the free surface of the mesothelial cells. At pH 1.5, the colloidal particles aggregated in a dotted fashion; in those stained at pH 7.0, the particles arranged in fine strands (100-300 nm in length). This difference may occur as a structural transformation due to pH level changes. The string like structure seemed to correspond well to membrane associated sialomucin. The urinary surface of the rat glomerular podocytes possessed negatively charged sites detectable with cationic colloidal iron even at pH 1.5. Neuraminidase and LFA treatments erased iron staining. Substance containing sialic acid such as podocalyxin on the podocyte surface may be stained. This study shows that negatively charged sites of the substance covering the free surface of these regions repulse each other to maintain the serosal cavities or the podocyte end feet slits. PMID- 11729981 TI - Cytoarchitecture of the bronchiolar wall in the rat lung: scanning electron microscopic studies. AB - The cytoarchitecture of rat bronchiolar walls was directly visualized from the adventitial side by scanning electron microscopy after removal of collagen and elastic fibers with a KOH-collagenase digestion method. The bronchioles were surrounded by cord-shaped smooth muscle fibers, which ran circularly to the long axis of the bronchiolar tree. Branches of the bronchial artery were located outside the bronchiolar smooth muscle layer, ran toward the periphery, gave off smaller branches one after another, and finally formed capillary networks both inside and outside the smooth muscle layer. Collecting venules arising from the capillary network ran independently of arterioles, but were gradually accompanied with arterioles as they became thickened. Peripheral nerves covered with a perineurial sheath were also present outside the bronchiolar smooth muscle layer; they gave off branches toward the periphery, and finally entered the bronchiolar smooth muscle layer. Lymphatic vessels were sometimes found at the bifurcation of bronchioles. These findings provide important basic data on the structure and functions of the bronchiolar wall. PMID- 11729982 TI - Functional implications of extensive new data on the innervation of pulmonary neuroepithelial bodies. AB - Pulmonary neuroepithelial bodies (NEBs) are innervated organoid groups of neuroendocrine cells, present in the epithelial lining of intrapulmonary airways of man and all air-breathing vertebrates studied so far. NEBs receive a vagal nodose sensory innervation that is considered by other authors as their main, if not their only innervation, although apparently not needed for the normal development and maintenance of NEBs. In the present study of NEBs in the developing rat lung (gestational day 16 - adult), immunoreactivity (IR) for calcitonin gene-related peptide (CGRP; a marker for NEBs), protein gene product 9.5 (PGP9.5; a marker for NEBs and neuronal elements), calbindin D28k (CB; a calcium binding protein), and for the growth-associated protein 43 (GAP-43; a marker for growing or remodelling nerve fibers) was combined with vagal denervation experiments. GAP-43 and CB IR revealed that the vagal sensory innervation of airways precedes the prenatal development of NEBs by several generations of branching. Unlike several other nerve fiber populations innervating NEBs, the vagal sensory component apparently does not express GAP-43 IR in mature lungs. GAP-43 labelling, however, did reveal newly ingrowing intraepithelial vagal sensory fibers, specifically reinnervating NEBs in adult rats 2 weeks following a cervical vagal crush. In conclusion, details of the innervation of pulmonary NEBs were disclosed that had been invisible so far, shedding new light on its complexity and probable involvement in the normal development and functional maintenance of NEBs throughout life. PMID- 11729983 TI - Structural and transport property alterations of the lung capillary endothelium in diabetes. AB - There are reports that lung function is altered in diabetes mellitus. Since data are limited concerning the structural--functional correlates in hyperglycemic animals, we designed experiments to assess: (i) whether hyperglycemia induces changes in the structure of the lung capillary endothelial cells (EC), and (ii) the effect of advanced glycation endproducts of albumin (AGE-Alb) on EC. Experiments were conducted on mice and hamsters rendered diabetics by streptozotocin (D-STZ) injection; age-matched animals were used as controls (C). The structure of EC, and the interaction of EC with AGE-Alb adsorbed to 5 nm colloidal gold (AGE-Alb.Au) were examined by electron microscopy; the uptake and cellular distribution of [125I]-AGE-Alb were investigated by spectrometry and autoradiography. The results showed that, compared to C group, in D-STZ animals the alveolar capillary EC exhibited: (i) an uneven distribution of the anionic sites exposed by the luminal plasmalemma, (ii) a metabolicaly active phenotype, (iii) a thickened basal lamina provided with focal nodules, similar in density and distribution to those found in the glomerular basal lamina in diabetes; in addition, (iv) a narrowed or collapsed lumen was found in approximately 30% of the capillaries. The functional alterations of lung EC in diabetes consisted in increased uptake of intravascularly infused [125I]-AGE-Alb and enhanced transcytosis and endocytosis of AGE-Alb.Au. The fusion of plasmalemmal vesicles, and opening of the interendothelial junctions observed, may account for the increased permeability of the alveolar capillary EC. This study asserts that hyperglycemia affects the structure and functional properties of the alveolar capillary EC, and suggests the existence of microangiopathic alterations in diabetic lung. PMID- 11729984 TI - The three-dimensional structure of the neonatal mouse kidney as revealed by scanning electron microscopy after KOH treatment. AB - The shape and arrangement of the developing nephrons were studied three dimensionally by scanning electron microscopy (SEM) of the neonatal mouse kidney. The specimens were treated with the KOH digestion method in order to remove extracellular connective tissue components, thus enabling the direct observation of the developing nephrons at various stages. At the subcapsular region of the renal cortex, the ureteric ducts were observed as branched tubules with terminal swellings or ampullae. Newly formed blood vessels were often associated with terminals of these ureteric ducts. The cup-shaped renal corpuscles had aggregations of mesangial cells with blood vessels in the groove. At the vascular pole of mature nephrons, extraglomerular mesangial cells were observed as a cellular sheet, which was continuous with the smooth muscle layer of afferent and efferent blood vessels. The present study also demonstrated the shape of the immature podocytes in relation to the endothelial morphology of glomerular capillaries. PMID- 11729985 TI - Morphogenetic activity of extracellular matrices on cultured podocytes. Laminin accelerates podocyte process formation in vitro. AB - Morphogenetic effects of various extracellular matrix proteins on the renal podocyte were investigated using the conditionally immortalized podocyte cell line. Podocytes were plated on glass coverslips and coated with the following matrix proteins: laminin-10/11, laminin-1, fibronectin, collagen type IV, collagen type I. Three hours after plating, podocytes on laminins developed prominent processes, while those on other matrix proteins started to elongate processes after two days. Vinculin-immunolabeling showed that podocytes plated on laminins possessed thin rod-shaped focal contacts, whereas those on fibronectin showed large dot-shaped focal contacts. Inhibition of serine/threonine protein kinases induced podocyte process formation in an extracellular matrix-independent manner. The present study reveals the significance of laminin on podocyte morphogenesis in vitro, and shows that different extracellular matrix proteins trigger different intracellular signals governing podocyte morphogenesis. Taken together with our previous studies, podocyte process formation is thought to be regulated by protein Ser/Thr phosphorylation. PMID- 11729986 TI - Ultrastructural study of mouse renal glomeruli under various hemodynamic conditions by an "in vivo cryotechnique". AB - Three-dimensional ultrastructure of mouse renal glomeruli under various hemodynamic conditions was studied by scanning electron microscopy with an "in vivo cryotechnique" followed by freeze-substitution. These results were also compared with those of conventionally fixed glomeruli at different perfusion pressures. Some kidneys of anesthetized mice were directly frozen in vivo under normal blood flow condition. Others were frozen in the same way after ligation of the lower abdominal aorta or their heart arrest. The frozen specimens were routinely freeze-substituted, freeze-dried and ion-sputtered for SEM. For another conventional fixation method, some kidneys were perfused with glutaraldehyde at pressures of 150 cm or 250 cm water, and other renal cortical tissues were routinely immersion-fixed with glutaraldehyde. In freeze-substituted specimens with the in vivo cryotechnique, interdigitating foot processes of podocytes exhibited smooth surface contours with less shrinkage and their surfaces were arranged more tightly than those seen by the conventional fixation method. In the anesthetized mice, filtration slits between the foot processes were found to be narrow after the heart arrest, but they were wide under the normal blood flow condition. After the aorta ligation, the filtration slits became wider and the foot processes were more elongated. The ultrastructure of capillary loops in functioning renal glomeruli were revealed to be dynamically changing in the living state, depending on various hemodynamic conditions. PMID- 11729987 TI - Using vital microscopy (tscm) to evaluate living kidneys. AB - Tandem Scanning Confocal Microscopy (TSCM) is a non-invasive form of vital microscopy which can be used to evaluate superficial uriniferous tubules in living kidneys. Because TSCM has a number of advantages over conventional microscopic examination of renal biopsies, the present study was undertaken to determine whether the histopathological images obtained by TSCM can be correlated with post-transplant renal function. The kidneys of New Zealand male rabbits were harvested, flushed in Euro-Collins solution, and stored at 0-2 degrees C for periods of 24, 48, 67 and 72 hours prior to transplantation. As expected, there was a significant deterioration in post-transplant renal function as the kidneys were stored for longer periods of time. TSCM observations of the kidneys prior to their transplantation revealed characteristic histopathological changes of the proximal convoluted tubules which correlated closely with post-transplant renal function. These observations support the proposed use of TSCM in evaluating human donor kidneys prior to their transplantation. PMID- 11729988 TI - Localization in situ of specific RNA by electron microscopy. AB - To better understand the metabolism of RNA in nuclei, the analysis of precise nuclear distribution of specific RNA would be essential. For this purpose, nonradioactive electron microscopic (EM) in situ hybridization may be the most appropriate technique while the details required for the technique have not been fully established. In the present study, we attempted to localize 28S and 18S rRNAs in the nuclei of mouse Sertoli cells by EM in situ hybridization as a model system. After various preliminary experiments we chose the pre-embedding method; fresh-frozen sections of mouse testis were fixed with a mixture of 4% paraformaldehyde and 0.1% glutaraldehyde, digested with 10 microg/ml of proteinase K and hybridized with thymine-thymine (T-T) dimerized oligodeoxynucleotides (oligo-DNA) complementary to a part of 28S and 18S rRNAs. Then, the T-T dimers were detected enzyme-immunohistochemically with horseradish peroxidase (HRP) labeled anti-T-T dimer. After osmification of HRP products, the sections were embedded in Epon resin, cut into 100 nm ultra-thin sections and observed under a transmission electron microscope. As a result, we successfully localized both 28S and 18S rRNAs in the dense fibrillar and granular components of the nucleolus, showing the usefulness of nonradioactive EM in situ hybridization in the nuclear localization of specific RNA. PMID- 11729989 TI - The quest for factors regulating the development of chick embryonic insulin cells in vitro. AB - A mesenchymal factor(s) is believed to modulate the development of the embryonic pancreas. However, the identity of this factor(s) remains elusive. Its importance in the development of the endocrine cells of the pancreas, particularly insulin cells, is considered. Our studies seek to identify factors that may influence insulin cell proliferation and differentiation, and hence may have positive implications for patients suffering from diabetes. Our experiments are based on the in vitro model which utilised the 5-day dorsal pancreatic bud of the chick embryo. These buds, devoid of most of their mesenchyme, are cultured on Matrigel in a serum-free medium for 7 days. We have investigated the effects of retinoic acid, retinoic acid in combination with IGF-1, nicotinamide and activin in this system. While most of these factors increased the proportion of insulin cells over that of their respective controls, as yet none have matched or exceeded the proportion of insulin cells demonstrated in vivo. PMID- 11729990 TI - Somatostatin- and Substance P-ergic neurons and Glial cell line-derived neurotrophic factor in the human archicortex. AB - By means of immunohistochemistry, the localization of Somatostatin (SOM)- and Substance P (SP)-ergic neuronal populations was compared to the occurrence of Glial cell line-derived neurotrophic factor (GDNF) in the human hippocampus from prenatal to adult life stages. The results obtained i) confirm previous reports on the distribution of SOM and SP; ii) show that GDNF-like immunoreactivity occurs in an ample population of hippocampal neurons, with a main location in the pyramidal cells; iii) identify regions of codistribution of either neuropeptide with GDNF-positive elements. Although coexistence of GDNF with SOM or SP was not detected, the possibility that the trophic factor may act on the neuropeptide containing neurons can be envisaged and is worth further analysis. PMID- 11729991 TI - Degranulation of mast cells in the rat thalamus. AB - Brain mast cells are selectively concentrated in the thalamus of many mammalian species. We here describe by light and electron microscopy in the normal thalamus of adult rats the features of mast cell degranulation, which indicate an active release of the mediators stored in their intracellular granules. The state of activity of thalamic mast cells in basal conditions was found to range from the release of a few granules to a massive degranulation, and the latter process was much less frequent than a partial degranulation. Mast cells were subdivided in three categories (fully granulated, partially or massively degranulated) on the basis of their cytoplasmic features revealed by acidic toluidine blue staining; the fully granulated cells were found to represent only 23 % of thalamic mast cells. This strategy of evaluation could be of help in the comparison of the functional correlates of mast cells in different conditions and experimental paradigms. However, we also demonstrated with image analysis a continuum of the variation of staining intensity of granulated and degranulating mast cells, without a sharp subdivision into different categories. Therefore our results reveal that the vast majority of mast cells are active in the thalamus in basal conditions, and that image analysis can provide an objective index of the activity of these cells. PMID- 11729992 TI - The extracellular matrix in the mature brain: its reactions to endo-alpha-N acetylgalactosaminidase and collagenase. AB - The hippocampal subiculum in the adult rat brain contains many neurons with nerve cell surface glycoproteins which are linked by collagenous ligands to perineuronal proteoglycans. The nerve cell surface glycoproteins or their terminal N-acetylgalactosamines are digested by endo-alpha-N acetylgalactosaminidase. The terminal N-acetylgalactosamines linked by the collagenous ligands are not digested by endo-alpha-N-acetylgalactosasminidase. The collagenous ligands associated with the terminal N-acetylgalactosamines were digested by collagenase. The newly exposed terminal N-acetylgalactosamines by this collagenase incubation were digested by endo-alpha-N-acetyl galactosaminidase. These findings on the rat agree with those obtained in our previous studies of the adult mouse brain samples. Furthermore they emphasize our hypothetical model that the perineuronal proteoglycans are related--via collagen ligands--with the nerve cell surface glycoproteins. PMID- 11729993 TI - Three-dimensional microanatomy of mechanoreceptors and their possible mechanism of sensory transduction. AB - The fine structure of sensory nerve endings and their topographical relationships with surrounding tissues were examined by a combination of scanning and transmission electron microscopy in order to analyze the mechanism of mechanoreception. Observations were reported on Ruffini endings in periodontal ligaments of rat incisors, and on longitudinal lanceolate endings surrounding rat sinus hairs. Both types of receptors exhibited the triplet structure known as the axon-Schwann cell complex; a flattened axon terminal was sandwiched between two Schwann cell lamellae. The two receptor types additionally revealed their specific modifications at each distal end, where fine tuft-like processes of Schwann cells projected into surrounding tissues with finger-like projections of an axon terminal attached to their bases. In the Ruffini endings of the periodontal ligament, the terminal tufts coiled about collagen bundles in favor of continuous transmission of tissue distortions to their accompanying axon fingers. In the lanceolate endings of sinus hair follicles, the Schwann cell tufts were suspended in an amorphous matrix with only their end feet anchored to rigid connective tissue elements. Terminal axon fingers associated with these Schwann cell processes were supposed to transiently deflect during acceleration and deceleration phases of a given hair movement because of inertia. The present study proposes the terminal tuft of Schwann cell processes and their accompanying axon fingers as a structural complex which potentially contributes to mechano electric transduction. PMID- 11729994 TI - Role of nitric oxide in the development of retinal projections. AB - Synthesis of nitric oxide (NO) occurs downstream from activation of NMDA receptors and NO acts as a retrograde messenger, influencing the refinement and stabilization of coactive afferent terminals. Cells and neuropil in the rat superior colliculus (SC) and lateral geniculate body (LGB) show intense, developmentally regulated activity for NO synthase (NOS). To study the role of NO in the development of retinogeniculate and retinotectal axon arbors, we examined primary visual projections of rats that had received daily i.p. injections of L NoArg (an NOS inhibitor) for 4-6 weeks starting from postnatal day 0. Retinal fibers labeled by intraocular injection of the B subunit of cholera toxin were revealed immunohistochemically and the density of fibers in the superficial SC and in the dorsal LGB was measured by computerized image analysis. Single retinocollicular terminal arbors were reconstructed at the computer (Neurolucida). Treated rats showed significant alterations in ipsilateral retinotectal projections, in the mediolateral and anteroposterior axes: there was an increase in the density of fibers entering the SC, in branch length, and in numbers of boutons on retinotectal arbors in the treated group. Ipsilaterally projecting retinal axons also showed an increase in density and distribution in the dorsal nucleus of the LGB. If animals were allowed to survive for several months after stopping treatment, similar changes were also noted, but these were much less striking. Our results suggest that, in the mammalian visual system, NO released from target neurons in the SC and LGB serves as a retrograde signal which feeds back on retinal afferents, influencing their growth. PMID- 11729995 TI - Synaptic ribbon dynamics in photoreceptors of mice. AB - The present study deals with structural plasticity of a special type of chemical synapse, the ribbon synapse. Near the presynaptic membrane ribbon synapses contain conspicuous electron-dense synaptic bodies which appear mainly as rod like profiles under the transmission electron microscope. In addition, club shaped and spherical profiles may be present, the function of which is unclear. To gain some insight into the significance of the latter structures we studied their presence in rod-type photoreceptor cells of BALB/c mice under different lighting conditions. Quantification revealed that the club-shaped and the spherical profiles showed a clear light/dark dependence. They were virtually absent at night and increased strikingly in number when the animals were exposed to light. When darkness was extended into the morning, the profiles remained low in number. As the rod cells diminish their neurotransmitter release during the light phase, the present findings are interpreted as signs of synapse inactivation. PMID- 11729996 TI - Membrana limitans interna and epiretinal membrane lying on macular holes. Some morphological observations. AB - The structure of the membrana limitans interna (MLI) in the region of the macula has been investigated by electron microscopy in (a) 2 enucleated human adult eyes and (b) 38 surgically removed samples associated with an epiretinal membrane (ERM). In the enucleated eye, the glia cells were vitrad bordered either by the lamina rara or, directly, by the lamina densa. Both extended into a coarse network whereby the lamina densa, through repeated branches and anastomoses, delimited large meshes, the lamina rara formed their contents. High magnification revealed that both meshes and contents of this network were composed by a further, finer network. It is suggested that strips and small openings of the finer network are homologous to the cords and intercordal spaces, respectively, which have been indicated as the common, basic structures of most of the basement membranes. The MLI excised with an ERM had the same structure. In some of the ERM associated with a macular hole, myofibroblasts prevailed among the cells. They showed indented nucleus, stress fibers abuting on the plasma membrane or in apparent continuity with bundles of extracellular filaments (microtendons), gap junctions. The cells lay on or were surrounded by a discontinuous basement membrane. PMID- 11729997 TI - Expression of SCC in ovarian granulosa cells and cultured cells, induced rapid structural changes in mitochondria. AB - In this study, the relation of P450scc expression and mitochondrial ultrastructure was examined in rat granulosa cells at the time of ovulation, and in the NIH/3T3 cells. Before ovulation in the ovary, granulosa cells of Graafian follicle which expressed only mRNA of P450scc had elongated mitochonria with lamellar cristae. After ovulation, granulosa lutein cells which expressed both P450scc mRNA and protein had oval and round mitochondria with tubular or vesicular cristae. Two different cytochrome P450scc cDNA fragments in length were subcloned into pEGFPN vector, transfected into NIH/3T3 cells, and the mitochondrial structure was examined under fluorescent microscope with GFP and by electron microscopy. 5'end of cytochrome P450scc contained mitochondrial localization signal, and was composed of about 40 amino acids. NIH/3T3 cells had filamentous and elongated mitochondria with lamellar cristae, free ribosomes, and rER. After the transfection of short fragment of SCC(scc-s:200bp), mitochondria remained filamentous and their cristae also remained lamellar. On the other hand, when almost full length of SCC fragment(scc-f:1.1kb) was transfected, globular and round mitochondria were labeled with GFP, and round or oval mitochondria with vesicular or tubular cristae could be examined by electron microscope. Our study suggests that cytochrome P450scc located in mitochondrial inner membrane plays an important role to determine the mitochondrial morphology in vivo and in vitro. PMID- 11729998 TI - Intracellular P-glycoprotein in multidrug resistant tumor cells. AB - The overexpression of the drug-efflux molecular pump P-glycoprotein (P-gp) may confer to tumor cells the multidrug resistant (MDR) phenotype, which is one of the causes of cancer chemotherapy failure. By investigating several in vitro models of human tumor cells, we observed that P-gp, in addition to its localization on the plasma membrane, can also be found intracellularly. In particular, by using immunocytochemical and cytofluorimetric methods, we revealed that in MDR breast cancer cells (MCF-7) a significant level of P-gp was expressed in the Golgi apparatus, which is the major site of accumulation of the antitumoral compound doxorubicin. Moreover, we demonstrated the intracellular location of P-gp in three stabilized human melanoma cell lines which had never undergone cytotoxic drug treatment and did not express the transporter molecule on the plasma membrane. Double immunofluorescence labelling and immunoelectron microscopy revealed, also in this tumor cell type, the location of P-gp in the Golgi apparatus where it seems to play a pivotal role in intracellular drug transport. Finally, we analyzed the expression, localization and function of drug transport proteins in human colon carcinoma lines (LoVo) exhibiting different degrees of intrinsic or drug-induced resistance. We found that only MDR LoVo cells expressed P-gp on the plasma membrane while both low-level drug resistant clonal LoVo cells and MDR LoVo cells appeared to be positive for intracellular P gp. Our findings suggest a functional role of the intracytoplasmic P-gp in the transport and sequestration of drugs. This represents a complementary protective mechanism of tumor cells against cytotoxic agents. PMID- 11729999 TI - Cell localization of the anti-inflammatory protein annexin 1 during experimental inflammatory response. AB - The localization of the glucocorticoid-inducible protein annexin 1 (ANX-1) in leukocytes during the process of experimental inflammation has been studied using immunocytochemistry. ANX- 1 immunoreactivity was detected in extravasated neutrophils and eosinophils as well as in resident tissue mast cells. Following injection of carrageenin, the mesenteric tissue was highly inflamed with large presence of leukocytes (predominantly neutrophils with a small percentage of eosinophils) adherent to post-capillary venules and extravasated in the perivascular tissue. ANX-1 immunoreactivity was detected in the cytosol of neutrophils and eosinophils mainly associated with granules and/or vesicles. A good degree of localization in the endosomes was observed in the neutrophils. In both cell types, some ANX-1 immunoreactivity in the nucleus and in the plasma membrane was also detected. Resident mast cells were also activated. Mast cells were positive for ANX-1, without apparent changes in protein content in relation to their activation status. Degranulated mast cells still presented ANX-1 associated with the granule matrix. In conclusion, this study demonstrated the presence of ANX-1 in leukocytes that play a central role in the host inflammatory response. These are the extravasating polymorphonuclear cells, or the resident mast cells. These data provide morphological support to the notion that endogenous and exogenous ANX-1 are able to modulate the reactivity of these cell types, and more generally, of the experimental inflammatory reaction. PMID- 11730000 TI - Autoschizis: another cell death for cancer cells induced by oxidative stress. AB - Scanning and transmission electron microscopy (SEM and TEM) were employed to characterize the cytotoxic effects of vitamin C (VC), Vitamin K3 (VK3) or a VC:VK3 combination on a human bladder carcinoma cell line (T24) following vitamin treatment. T24 cells exposed to VC alone showed membrane defects. VK3-treated cells show greater damage than VC treated cells because they exhibit membrane defects, cytoskeletal damage, excision of cytoplasm, and a substantial decrease in the number of viable cells. VC: VK3 treatment exacerbates the damages, especially intranuclear and nucleolar and induces an extreme reduction of cell size by cytoplasmic self-excision. Conversely, the nuclear envelope remains intact and the rough endoplasmic reticulum (RER) maintains its integrity until karyorrhexis occurs through a new phenomenon of cell death that we have named "autoschizis". From our morphological studies and previous biochemical reports on the topic, we are able to propose that this autoschizic cell death found is induced by oxidative stress. PMID- 11730001 TI - Chromatin fibers organization within the nucleus. AB - In order to evaluate at the ultrastructural level the three dimensional arrangement of the dispersed chromatin during the intephase, the immunogold detection of Bromodeoxyuridine (BrdU), of the DNA polymerase alpha and of the proliferating cell nuclear antigen (PCNA) was performed on human HL60 leukemia cells and nuclear matrices extracted from the same cellular model. The Field Emission In lens Scanning Electron Microscopy analysis of the ultrathin cryosectioned cells revealed the presence of a chromatin three dimensional network where the different constituents appeared repetitively assembled. Also the nuclear matrix showed a repetitive structure, on which the deprivation of the DNA corresponded to the selective loss of particular class sized fibers. The single or multiple combined immunolocalization of different structures involved in the DNA replication, where BrdU, DNA polymerase alpha and PCNA represent, respectively, the substratum, the polymerizing enzyme and a regulator of the reaction, allowed the understanding of its reciprocal spatial relationship on the dispersed interphasic chromatin and the role of the nuclear matrix in the replicative process. PMID- 11730002 TI - Beta-amyloid fibrils of Alzheimer's disease: pathologically altered, basement membrane-associated microfibrils? AB - Beta amyloid fibrils were examined in situ in the cerebral cortex of brains from patients with Alzheimer's disease using high resolution ultrastructural and immunohistochemical techniques. The main body of the fibril was identical with that of microfibrils and was made up of a core containing amyloid P component (AP), and a surface layer. Beta amyloid protein (Abeta) in the form of 1 nm wide flexible filaments was associated with the external surface of the microfibril. In cerebrovascular amyloid angiopathy the fibrils were formed at the outer surface of the vascular basement membrane. Overproduction of microfibrils has been reported at the basement membrane of "leaky" capillaries including the glomerular capillary in disease or leaky alveolar-capillary walls of normal lungs. Similarly, in Alzheimer's disease overproduction of microfibril-like beta amyloid fibrils in amyloid angiopathy coincided with breakdown of the blood-brain barrier of the cerebromicrovasculature. Thus, in the above three locations, the presence of abundant microfibrils, or microfibril-like structures, may be related to plasma which leaks out of the circulation into the adjoining vascular basement membrane. AP is an essential constituent of microfibrils and since the only site where AP is available in the cerebral cortex is in leaky microvasculature, a chronic, steady supply of AP into perivascular areas may be the cause of overproduction of microfibrils. Brain "microfibrils" may further be altered pathologically into beta amyloid fibrils by the addition of Abeta. The origin of the fibrils in senile plaques may also be the microvasculature since in the area of the plaques no source of AP is apparent. PMID- 11730003 TI - George Emil Palade and Don Wayne Fawcett and the development of modern anatomy, histology and contemporary cell biology. PMID- 11730004 TI - Partial restoration of degraded DNA from archival paraffin-embedded tissues. PMID- 11730005 TI - Sodium bisulfite analysis of the methylation status of DNA from small portions of paraffin slides. PMID- 11730006 TI - Efficient transformation of Halobacterium salinarum by a "freeze and thaw" technique. PMID- 11730007 TI - PCR primers and conditions for nonmarine ostracods. PMID- 11730008 TI - Use of coisogenic host blastocysts for efficient establishment of germline chimeras with C57BL/6J ES cell lines. AB - Gene targeting in embryonic stem (ES) cells allows the production of mice with specified genetic mutations. Currently, germline-competent ES cell lines are available from only a limited number of mouse strains, and inappropriate ES cell/host blastocyst combinations often restrict the efficient production of gene targeted mice. Here, we describe the derivation of C57BL/6J (B6) ES lines and compare the effectiveness of two host blastocyst donors, FVB/NJ (FVB) and the coisogenic strain C57BL/6-Tyr(c)-2J (c2J), for the production of germline chimeras. We found that when B6 ES cells were injected into c2J host blastocysts, a high rate of coat-color chimerism was detected, and germline transmission could be obtained with few blastocyst injections. In all but one case, highly chimeric mice transmitted to 100% of their offspring. The injection of B6 ES cells into FVB blastocysts produced some chimeric mice. However; the proportion of coat color chimerism was low, with many more blastocyst injections required to generate chimeras capable of germline transmission. Our data support the use of the coisogenic albino host strain, c2J, for the generation of germline-competent chimeric mice when using B6 ES cells. PMID- 11730009 TI - Transfection-mediated cell synchronization: acceleration of G1-S phase transition by gamma irradiation. AB - We have previously provided evidence that the uptake of DNA into cells is cell cycle specific following transfection. We show here that, immediately after transfection, successfully transfected cells are greatly enriched for cells in early G1 or G0 phase and that, upon removal of the DNA precipitates, cells progress through G1 and enter S phase in a synchronous fashion. We also demonstrate that this approach can be utilized in meaningful cell-cycle experiments, and we show that gamma irradiation accelerates the G1-S phase transition in a cell line with a functionally inactive p53 protein. PMID- 11730010 TI - Cre stoplight: a red/green fluorescent reporter of Cre recombinase expression in living cells. AB - The Cre/lox system is a powerful genetic tool with which to manipulate the genome. Here, we describe the development of a simple reporter system for Cre recombinase, called the Cre Stoplight. In the absence of Cre, the red fluorescent protein is expressed; when Cre catalyzes a recombination event, the green fluorescent protein is produced. Testing this system in transiently transfected cells showed that it produced robust signals (90% of the cells converted from red to green) when equal amounts of the plasmids encoding Cre recombinase and the Cre Stoplight were used. A 1:100 ratio of enzyme to reporter plasmid produced similar results, and a 1:10000 ratio was necessary to significantly reduce the number of cells converting to green (1%). PMID- 11730011 TI - Removal of polyA tails from full-length cDNA libraries for high-efficiency sequencing. AB - We have developed a method to overcome sequencing problems caused by the presence of homopolymer stretches, such as polyA/T, in cDNA libraries. PolyA tails are shortened by cleaving before cDNA cloning with type IIS restriction enzymes, such as GsuI, placed next to the oligo-dT used to prime the polyA tails of mRNAs. We constructed four rice Cap-Trapper-selected, full-length normalized cDNA libraries, of which the average residual polyA tail was 4 bases or shorter in most of the clones analyzed Because of the removal of homopolymeric stretches, libraries prepared with this method can be used for direct sequencing and transcriptional sequencing without the slippage observed for libraries prepared with currently available methods, thus improving sequencing accuracy, operations, and throughput. PMID- 11730012 TI - pAd5-Blue: direct ligation system for engineering recombinant adenovirus constructs. AB - We have engineered a new vector that makes use of direct ligation for the generation of replication-defective recombinant adenovirus constructs. In the pAd5-Blue vector, unique yet common restriction endonuclease sites exist, that allow cloning in a directional manner of a gene of interest under control of a cytomegalovirus promoter, upstream of a simian virus 40 polyadenylation signal. The insertion of the new gene replaces the beta-galactosidase alpha gene fragment in the pAd5-Blue vector, allowing the identification of recombinants in bacterial culture by the selection of white colonies. Plasmid DNA from white colonies is digested with PacI and transfected into 293 cells, resulting in the generation of a homogenous population of adenovirus containing the gene of interest. The pAd5 Blue vector system does not rely on recombination either in mammalian or bacterial cells. Furthermore, because of compatible overhangs, the variety of restriction endonucleases that can be used to generate the inserted gene gives flexibility to the process for greater ease of use. The system is quick and straightforward, allowing the generation of recombinant adenoviruses within three weeks of obtaining an appropriate insert. This new vector should greatly enhance the ease and speed with which new recombinant adenovirus constructs can be made. PMID- 11730013 TI - Electrophoretic mobility shift scanning using an automated infrared DNA sequencer. AB - Electrophoretic mobility shift assay (EMSA) is widely used in the study of sequence-specific DNA-binding proteins, including transcription factors and mismatch binding proteins. We have established a non-radioisotope-based protocol for EMSA that features an automated DNA sequencer with an infrared fluorescent dye (IRDye) detection unit. Our modification of the elec- trophoresis unit, which includes cooling the gel plates with a reduced well-to-read length, has made it possible to detect shifted bands within 1 h. Further, we have developed a rapid ligation-based method for generating IRDye-labeled probes with an approximately 60% cost reduction. This method has the advantages of real-time scanning, stability of labeled probes, and better safety associated with nonradioactive methods of detection. Analysis of a promoter from an industrially important filamentous fungus, Aspergillus oryzae, in a prototype experiment revealed that the method we describe has potential for use in systematic scanning and identification of the functionally important elements to which cellular factors bind in a sequence-specific manner. PMID- 11730014 TI - New E. coli cloning vector using a cellulase gene (celA) as a screening marker. AB - The extracellular endoglucanase A gene of Clostridium thermocellum (celA) was used as a screening marker for E. coli cloning vector A 1.4-kb EcoRI fragment containing celA from pTvec/celA was isolated and cloned into a pUC18 deleting beta-galactosidase gene fragment. The constructed vectors, pCEL1, pCEL10, pCEL11, and pCEL20, have different multiple cloning sites within celA. If the cellulase, CelA, is inactivated by insertion of a foreign DNA fragment into multiple cloning sites, the recombinant transformants show no clear halos on an agar plate containing cellulose. This process overcomes the ambiguity of color screening in the X-gal/beta-galactosidase system, and over 90% of the recombinant transformants with no halos have foreign DNA inserts. Several E. coli strains were transformed successfully with pCEL series vectors regardless of mutation for alpha-complementation. Because E. coli strains do not have a cellulase gene, a vector using a cellulase gene screening marker can be used in any E. coli strain without limit. The new cloning system is very efficient, convenient, and cost effective. PMID- 11730015 TI - A workingperson's guide to deconvolution in light microscopy. AB - Thefluorescence microscope is routinely used to study cellular structure in many biomedical research laboratories and is increasingly used as a quantitative assay system for cellular dynamics. One of the major causes of image degradation in the fluorescence microscope is blurring. Deconvolution algorithms use a model of the microscope imaging process to either subtract or reassign out-of-focus blur. A variety of algorithms are now commercially available, each with its own characteristic advantages and disadvantages. In this article, we review the imaging process in the fluorescence microscope and then discuss how the various deconvolution methods work. Finally, we provide a summary of practical tips for using deconvolution and discuss imaging artifacts and how to minimize them. PMID- 11730016 TI - Chemiluminescence microscopy as a tool in biomedical research. AB - Chemiluminescence has become a standard tool in biomedical research. Chemiluminescent probes are used for immunoassays, nucleic acid identification, reporter gene assays, measuring enzyme activity, and the detection of ions and small molecules such as Ca2+, ATP, NO, O2- and H2O2. Along with the development of new chemiluminescent probes, significant progress has been made in techniques to measure chemiluminescence. Ultra-sensitive photometers or luminometers have become widely available and can be obtained with automatic injectors and microplate readers. In addition, imaging photon detectors have been developed that allow the imaging of chemiluminescence from gels, blots, and microplates. Imaging photon detectors have also been attached to microscopes and allow imaging of chemiluminescent probes and reporter genes in cells and tissues. Specific methods of photon collection, storage, and analysis have been developed for microscopic imaging of chemiluminescence. Two of these methods are discussed in detail. The first is a method of data storage that allows days of continuous imaging without creating oversized files. The second is a method for calibrating photon imaging microscopes using a low-light standard. Such calibration will be helpful for comparing the performance of various photon imaging systems and for comparing data obtained in different laboratories. PMID- 11730018 TI - Interrogation of multimeric DNA amplification products by competitive primer extension using bst DNA polymerase (large fragment). AB - Linear dsDNA composed of tandem repeats may be exponentially amplified by the strongly strand-displacing Bst DNA polymerase (large fragment) and two primers specific for opposite strands. When the repetitive DNA is derivedfrom rolling circle replication of a circular template, the reaction is termed cascade rolling circle amplification (CRCA). We have developed a variant of CRCA in which one primer is attached to the surface of a microwell and the other is labeled, thus enabling detection of amplified material using an ELISA-like protocol. The circular template is derived by annealing and ligation of a padlock on target DNA. It was found that there was good correlation between the synthesis of amplified material and signal. The specificity of the reaction with respect to single-nucleotide polymorphisms was investigated, and it was found that Bst DNA polymerase is prone to extension from primers with mismatched 3' ends. Reliable single nucleotide specificity was only obtained when pre-synthesized amplified material was interrogated by competitive primer extension. PMID- 11730017 TI - DNA probes using fluorescence resonance energy transfer (FRET): designs and applications. AB - Fluorescence resonance energy transfer (FRET) is widely used in biomedical research as a reporter method. Oligonucleotides with a DNA backbone and one or several chromophore tags have found multiple applications as FRET probes. They are especially advantageous for the real-time monitoring of biochemical reactions and in vivo studies. This paper reviews the design and applications of various DNA-based probes that use FRET The approaches used in the design of new DNA FRET probes are discussed. PMID- 11730019 TI - Automated real-time measurement of chemotactic cell motility. AB - We have developed a novel method, (ECIS/taxis), for monitoring cell movement in response to chemotactic and chemokinetic factors. In this system, cells migrate in an under-agarose environment, and their positions are monitored using the electric cell-substrate impedance sensor technology to measure the impedance change at a target electrode, that is lithographed onto the substrate, as the cells arrive at the target. In the studies reported here, Dictyostelium discoideum was used as a prototypical, motile eukaryotic cell. Using the ECIS/taxis system, the arrival of cells at the target electrode was proportional to the dose offolate used to stimulate the cells and could be assessed by changes in resistance at the electrode. ECIS/taxis was readily able to distinguish between wild-type cells and a mutant that is deficient in its chemotactic response. Finally, we have shown that an agent that interferes with chemotactic motility leads to the delayed arrival of cells at the target electrode. The multi well assay configuration allows for simultaneous automated screening of many samples for chemotactic or anti-chemotactic activity. This assay system is compatible with measurements of mammalian cell movement and should be valuable in the assessment of both agonists and antagonists of cell movement. PMID- 11730020 TI - Under-agarose folate chemotaxis of Dictyostelium discoideum amoebae in permissive and mechanically inhibited conditions. AB - Under-agarose chemotaxis has been used previously to assess the ability of neutrophils to respond to gradients of chemoattractant. We have adapted this assay to the chemotactic movement of Dictyostelium amoebae in response to folic acid. Troughs are used instead of wells to increase the area along which the cells can be visualized and to create a uniform front of moving cells. Imaging the transition zone where the cells first encounter the agarose, we find that the cells move perpendicular to the gradient and periodically manage to squeeze under the agarose and move up the gradient. As cells exit the troughs, their cross sectional area increases as the cells become flattened. Three-dimensional reconstruction of confocal optical sections through GFP-labeled cells demonstrates that the increase in cross-sectional area is due to the flattening of the cells. Since the cells locally deform the agarose and become deformed by it, the concentration of the agarose, and therefore its stiffness, should affect the ability of the cells to migrate. Consistent with this hypothesis, cells in 0.5% agarose move faster and are less flat than cells under 2% agarose. Cells do not exit the troughs and move under 3% agarose at all. Therefore, this assay can be used to compare and quantify the ability of different cell types or mutant cell lines to move in a restrictive environment. PMID- 11730021 TI - High-pressure gel loader for capillary array electrophoresis microchannel plates. AB - Microfabricated capillary array electrophoresis (microCAE) microchannel plates are the next generation of bioanalytical separation devices. To fully exploit the capabilities of microCAE devices, supporting technology such as robotic sample loading, gel loading, microplate washing, and data analysis must be developed. Here, we describe a device for loading gel into radial capillary array electrophoresis microplates and for plate washing and drying. The microplates are locked into a loading module, and high-pressure helium is used to drive aqueous separation media or wash solutions into the microchannels through fixtures connected to the central anode reservoir. Microplates are rapidly (30 s to 5 min) loaded with separation media, such as 3%-4.8% linear polyacrylamide or 0.7%-3.0% hydroxyethyl cellulose, for electrophoresis. The effective and rapid gel-filling and plate-cleaning methods together with short electrophoretic analysis times (2 30 min) make microCAE systems versatile and powerful nucleic acid analysis platforms. PMID- 11730022 TI - Genotyping of Cre-lox mice and detection of tissue-specific recombination by multiplex PCR. AB - Conditional gene targeting, based on Cre-lox or other systems, requires frequent genotyping of transgenic mouse populations and monitoring of tissue-specific Cre recombinatory efficiency. This is currently achieved by Southern analysis from tail- and tissue-derived DNA. Multiplex PCR amplification of the floxed (flanked by loxP sites) genomic region, combined with the PCR detection of the Cre transgene, simplifies this task. Here, we show that complete genotyping of a floxed locus is possible with three appropriately placed primers and that this triplex PCR can be performed simultaneously with a universal PCR assay for the detection of Cre transgenes. Using this approach, we also determined the ratios of recombined versus non-recombined floxed genomic segments in genomic DNA samples. This allowed us to estimate the efficiency of in vivo conditional inactivation from biopsy material and tissue samples that were too small for Southern analysis. As many new conditional knockouts are spatiotemporally restricted, such assays will become increasingly useful. The proposed PCR strategy is flexible and may be adapted to the structural specificities of any target gene. PMID- 11730023 TI - Drug design--chemistry and biology. PMID- 11730024 TI - Luminometric assay of platelet activation in 96-well microplate. AB - As of today, no practical method for large-scale functional anti-thrombosis agent screening exists. Based on the phenomenon that platelet activation results in the release of ATP from dense granules, we report the development and optimization of a 96-well microplate luciferase assay to assess platelet activation via luminescence detection of the released ATP. In addition, the assessment of re calcification-induced clotting of citrated platelet-rich plasma (PRP) is also possible. Collagen, thrombin, U46619, and ADP were shown to induce platelet activation in a concentration- and time-dependent manner The assay is applicable to PRP, washed platelets, and whole blood. Fundamentally, this is an ideal protocol for screening large numbers of anti-thrombotic drugs because of its sensitivity and the low amount of platelets required to detect simultaneous platelet activation. PMID- 11730025 TI - Hybridization cross-reactivity within homologous gene families on glass cDNA microarrays. AB - Glass cDNA microarrays can be used to profile the expression of thousands of gene targets in a single experiment. However, the potential for hybridization cross reactivity needs to be considered when interpreting the results. Here, we describe hybridization experiments with a model array representing four distinct functional classes (families): chemokines, cytochrome P-450 isozymes, G proteins, and proteases. The cDNA clones selected for this array exhibited pairwise sequence identities ranging from 55% to 100%, as determined by a homology scoring algorithm (LALIGN). Targets for microarraying were amplified by PCR and spotted in 4-fold replication for signal averaging. One designated target from each family was further amplified by PCR to incorporate a T7 promoter sequence for the production of synthetic RNA transcripts. These transcripts were used to generate fluorescent hybridization probes by reverse transcription at varying input concentrations. As expected, hybridization signals were highest at the matching target elements. Targets containing less than 80% sequence identity relative to the hybridization probe sequences showed cross-reactivities ranging from 0.6% to 12%. Targets containing greater than 80% identity showed higher cross reactivities (26%-57%). These cross-reactive signals were analyzed for statistical correlation with the length of sequence overlap, percent sequence identity, and homology score determined by LALIGN. Overall, percent sequence identity was the best predictor of hybridization cross-reactivity. These results provide useful guidelines for interpreting glass cDNA microarray data. PMID- 11730026 TI - In vitro assay for site-specific proteases using bead-attached GFP substrate. AB - Site-specific proteases, which catalyze cleavage of peptide bonds in specific amino acid sequences of target proteins, play important roles in various biological events of many living organisms. In humans, disruption in regulation of these site-specific proteases can lead to pathological consequences. Here, we report a simple in vitro assay for enzymatic activities of site-specific proteases. This assay system employs a protein substrate molecule that is comprised of (i) His-tag binding module, (ii) cleavage sites, and (iii) green fluorescent protein (GFP) detection module. In this study, prostate-specific antigen (PSA) and Thrombin-specific cleavage sites were introduced into the substrate molecules. The overexpressed GFP substrate protein was purified with the aid of Ni++-charged magnetic beads. On cleavage by either PSA or Thrombin, GFP was released from the bound magnetic beads, enabling a direct measurement of the cleaved product by fluorescence. Detection sensitivity, as well as the kinetics of reaction of PSA cleavage with the GFP substrate, was similar or better than commercially available PSA fluorogenic peptide substrate. This bead attached GFP substrate was also used for an inhibition assay using a competitive inhibitor of Thrombin. In conclusion, this assay offers a simple fluorescent method for monitoring the activity of the site-specific proteases. Furthermore, this system provides flexible means of incorporating varying sizes of flanking sequences adjacent to the cleavage site, which can be essential for studying the regulatory macromolecular interactions between proteases and their substrates. PMID- 11730027 TI - Convenient and rapid ribonuclease protection assay for use with primary cell cultures. PMID- 11730028 TI - An E-box in pGL3 reporter vectors precludes their use for the study of sterol regulatory element-binding proteins. PMID- 11730029 TI - Generalized 96-well format for quantitative and qualitative monitoring of altered protein expression and posttranslational modification in cells. PMID- 11730030 TI - What's new in the treatment of vitiligo. PMID- 11730031 TI - The cost of psoriasis treatment. PMID- 11730032 TI - Cost of caring for hospital-based patients in dermatology in Europe. PMID- 11730033 TI - Asymmetric periflexural exanthem of childhood: who are you? PMID- 11730034 TI - Contact allergy to corticosteroids: a challenge for dermatologists. PMID- 11730035 TI - Prurigo: where are you? PMID- 11730036 TI - Radiation dermatitis today. PMID- 11730037 TI - What causes acanthosis nigricans? PMID- 11730038 TI - Elastic fibre abnormalities in skin disorders: what's new? PMID- 11730039 TI - Morphometric analysis of elastic skin fibres from patients with: cutis laxa, anetoderma, pseudoxanthoma elasticum, and Buschke-Ollendorff and Williams-Beuren syndromes. AB - Computed morphometric analysis of elastic skin fibres in patients with cutis laxa, anetoderma, Williams-Beuren syndrome, pseudoxanthoma elasticum (PXE), and Buschke-Ollendorff syndrome, all clinically ascertained, was performed and compared with data obtained from healthy individuals of the same age. The diameters, area fractions (AA%) and volume fractions (VV%) occupied by pre elastic fibres and dermal elastic fibres were determined. Irrespective of age the diameter of dermal elastic fibres followed a Gaussian distribution for all groups studied. These diameters were taken into consideration for VV% determinations. Compared with data from skin of healthy subjects of similar age range, VV% of pre elastic fibres was significantly decreased in patients with cutis laxa, anetoderma, Williams-Beuren syndrome, and PXE and undetectable in Buschke Ollendorff patients. VV% of dermal elastic fibres was four- to fivefold increased in Buschke-Ollendorff syndrome, two- to threefold increased in PXE skin, four- to fivefold decreased in cutis laxa and anetoderma skin and about twofold decreased in Williams-Beuren skin. The diameter of oxytalan fibres was decreased in anetoderma and Williams-Beuren syndrome while oxytalan fibre diameter was unchanged in PXE and cutis laxa. The diameter of dermal elastic fibres was increased in PXE and Buschke-Ollendorff syndrome, but was decreased in anetoderma and Williams-Beuren syndrome and unchanged in cutis laxa. We demonstrated that cutis laxa, anetoderma, Williams-Beuren syndrome, PXE, and Buschke-Ollendorff syndrome could be easily differentiated by morphometric analysis of elastic skin fibres. Thus we propose that morphometric analyses together with skin biopsies are a valuable tool for distinguishing between inherited and/or acquired skin diseases known to display alterations of elastic fibres. PMID- 11730040 TI - Prevalence of superficial fungal infections among sports-active individuals: results from the Achilles survey, a review of the literature. AB - OBJECTIVE: To examine the effect of sport activities on the prevalence of foot disease in the Achilles survey, conducted during the spring of 1997 and 1998 in several European countries. SUBJECT: Foot diseases, especially fungal infections (tinea pedis and onychomycosis). METHODS: A questionnaire and a clinical examination regarding individuals presenting to a general practitioner for disorders irrespective of possible foot problems. RESULTS: More than 50% of subjects visiting a general practitioner had clinical evidence of foot disease, and approximately two-thirds of these had clinical evidence of a superficial fungal infection. The survey evidenced a significant age-dependent association between sporting activities and the prevalence of foot diseases and superficial fungal infection, especially in children. CONCLUSIONS: The results of this pan European survey indicate that sporting activities can have an unfavourable effect on the individual regarding the occurrence of foot disease and superficial fungal infection. The results indicate a need to pay more attention to foot disease, to predict and prevent future diseases and further complications. PMID- 11730041 TI - Bullous pemphigoid in Liguria: a 2-year survey. AB - BACKGROUND: The epidemiology of bullous pemphigoid (BP) is not clear because of the heterogeneity of the disease, and its possible association with internal malignancies has been under debate for many years. We report the findings of a 2 year study on incident BP cases in the Liguria region of Italy. SUBJECTS AND METHODS: Thirty-two patients with BP were collected over the 2-year period. Diagnosis was made based on clinical findings and confirmed by histology, direct immunofluorescence (DIF) and indirect immunofluorescence (IIF) with salt-split skin and monkey oesophagus, and immunoblotting (IB). All patients were thoroughly investigated for possible malignancies and all were followed up for 6 months to monitor the response to treatment. RESULTS: DIF showed linear deposits at the dermoepidermal junction in all but one patient. IIF gave positive findings for 15 sera tested with monkey oesophagus and 20 tested with salt-split skin. IB gave positive findings in 19 cases. There was a malignancy in six cases, but no clinical or immunological features that could be considered to predict this occurrence. CONCLUSIONS: The findings of this study are in accordance with most of the data found in the literature, including the fact that IgG serum levels did not predict the course of the disease. Contrary to previous indications, IgE levels were not indicative of disease course either. Mucosal lesions, erythema multiform-like lesions, negative IIF findings and antibodies to AgPB2 were not a prediction for the development of malignancy. PMID- 11730042 TI - The cost of hospital-related care of patients with psoriasis in Italy based on the AISP study. Associazione Italiana Studi Psoriasi. AB - The objective of this study was to assess the cost of caring for patients with psoriasis in Italy according to the AISP study (Associazione Italiana Studi Psoriasi or Italian Association for Studies on Psoriasis), involving 104 university and hospital centres and 7992 patients in 1994. The mean yearly cost of care for a single patient was calculated at 905 Euros. Hospitalization accounted for more than four-fifths of the costs, therapy for about one-eighth (systemic therapies were the most expensive) and office visits and day hospitals for the remainder. In our study series less than 20% of patients accounted for more than 90% of the total costs. PMID- 11730043 TI - Acanthosis nigricans: a new cutaneous sign in severe atopic dermatitis and Down syndrome. AB - Acanthosis nigricans (AN) occurs associated with many different systemic diseases, such as endocrine disorders and internal malignant neoplasms. To our knowledge, the association of AN with severe atopic dermatitis (AD) or Down syndrome has not been described before. This 82-month retrospective study included 1038 patients: AN was present in 4.9% of atopic patients and 50.9% of subjects with Down syndrome. AN was more frequent in patients with severe AD and in 100% of cases of hand dermatitis and juvenile plantar dermatosis, located on the interphalangeal and metacarpophalangeal joints, whereas in Down syndrome other flexures were also affected. The pathogenesis of AN in AD is unknown, but in Down syndrome it seems to be related to obesity. Possible insulin resistance underlyng the pathogenesis of AN in these patients is still unknown. PMID- 11730044 TI - Multiple myeloma with skin involvement. AB - Multiple myeloma (MM) is a plasma cell dyscrasia seen in the elderly that constitutes 10% of all haematopoietic neoplasias. It is a systemic disorder affecting various organs, in particular the kidneys and bones. Skin involvement is not a common finding. This report presents a case of MM with skin involvement. PMID- 11730045 TI - Waardenburg syndrome. AB - Waardenburg syndrome (WS) is caused by autosomal dominant mutations, and is characterized by pigmentary anomalies and various defects of neural crest derived tissues. We report a very interesting case of type 1 WS (WS 1) in an adult who presented all the symptoms characteristic of this syndrome. One particularly important clinical feature of WS is congenital hearing loss, which may severely handicap a child. A careful clinical description is useful to differentiate between various types of WS and other associated auditory-pigmentary syndromes. Type WS 1, characterized by dystopia canthorum, is caused by loss of function mutations in the PAX3 gene. PMID- 11730046 TI - Acral necrosis by Stenotrophomonas maltophilia. AB - BACKGROUND: Stenotrophomonas maltophilia (SM) has been considered a nosocomial pathogen. Nevertheless, community acquired infection may occur more frequently than usually recognized. CASE: We describe distal necrosis of the fingers by SM in a farmer, contracted in the community and successfully treated with a combination of cotrimoxazole and ciprofloxacin. The patient was diagnosed with chronic lymphocytic leukaemia 6 months later. CONCLUSIONS: This unusual presentation shows that infection with SM should be included in the differential diagnosis of the skin and soft tissue infection, even in apparently healthy patients. PMID- 11730047 TI - Acneiform eruption caused by amineptine. A case report and review of the literature. AB - Acne caused by amineptine has always been described with typical characteristic clinical features, and the retentional and cutaneous lesions are dose related. We present a case of acne-like eruption due to amineptine in a woman under treatment for chronic depression. PMID- 11730048 TI - Juvenile papular-purpuric gloves and socks syndrome. AB - Papular-purpuric gloves and socks syndrome (PPGSS) occurs mainly in young adults, but it should also be considered in the differential diagnosis of childhood exanthems. The case presented here is the youngest patient hitherto reported with PPGSS and supports the concept of a viral genesis. PMID- 11730049 TI - Crohn's disease presenting as a breast abscess: a case report. AB - A case of cutaneous Crohn's disease (CD) involving the areola-nipple region of both breasts in a 45-year-old woman is reported. The lesion had initially been diagnosed and treated as a simple abscess. Histopathological examination, however, showed granulomatous inflammation with eosinophils suggesting extraintestinal CD. The patient also had a minor area of cutaneous CD localized at the umbilicus and severe anogenital lesions. Twenty-six years previously the woman had undergone a course of sulphasalazine treatment together with steroid enema for chronic inflammatory colitis, but the colitis had been inactive for 25 years. Treatment with sulphasalazine, metronidazole, prednisolone and azothioprine had only minor effect on the skin lesions. CD is a systemic disorder and the most prominent manifestations may be extraintestinal. PMID- 11730050 TI - Brooke-Spiegler syndrome. AB - The Brooke-Spiegler syndrome is an autosomal dominant one characterized by cylindromas, trichoepitheliomas and occasionally spiradenomas. Within a given family, some members may have cylindromas whereas others may have trichoepitheliomas or both. We describe the coexistence of trichoepithelioma papulosum multiplex (also known as epithelioma adenoides cysticum of Brooke) and cylindromas in a 30-year-old man, and discuss the relationship between these two autosomal dominant syndromes. PMID- 11730051 TI - Polyvalent contact allergy to corticosteroids: a report of two cases. AB - Two cases of positive allergic reactions to eight patch-tested corticosteroid substances are reported. The patients were middle-aged women with a long history of contact dermatitis who had used topical corticosteroids for years. Pure corticosteroid substances were tested in therapeutic and 1% concentrations in ethanol and petrolatum. The intensity of reactions was different depending on the vehicle and concentration. Patients with hypersensitivity to several corticosteroid substances represent an important therapeutic problem. PMID- 11730052 TI - Vesicular prurigo pigmentosa cured by minocycline. AB - We present a case of prurigo pigmentosa associated with vesicles that we call 'vesicular prurigo pigmentosa'. The subject was treated using minocycline with good results and no recurrence of the lesions over a 2-year period. PMID- 11730053 TI - Localized vitiligo successfully treated with cream-psoralen + ultraviolet A. PMID- 11730054 TI - Diagnostic signs of cutaneous lymphomas. PMID- 11730055 TI - 'Tin-tack' sign in a patient with cutaneous B-cell lymphoma. PMID- 11730056 TI - Glucocorticoid action on skin collagen: overview on clinical significance and consequences. PMID- 11730057 TI - Localized monilethrix with improvement after treatment of iron deficiency anaemia. PMID- 11730058 TI - Lymphoedema associated with Kaposi's sarcoma. PMID- 11730059 TI - Raynaud symptoms as principal signs in a case of Sneddon's syndrome. PMID- 11730060 TI - Primary cutaneous anaplastic large cell lymphoma in a young girl. PMID- 11730061 TI - Response of chronic bullous dermatosis of childhood to a combination of dapsone and nicotinamide. PMID- 11730062 TI - Dermatitis artefacta. PMID- 11730063 TI - Angiosarcoma of the scalp: new treatment modalities. PMID- 11730064 TI - Sun-protective clothes: accuracy of laboratory testing. PMID- 11730065 TI - Asymmetric periflexural exanthem of childhood in an adult patient with parvovirus B19. PMID- 11730066 TI - Atypical tinea corporis caused by Microsporum gypseum in a subject with acquired immune deficiency syndrome. PMID- 11730067 TI - Tinea capitis in two young women: possible favouring role of hair styling products. PMID- 11730068 TI - Unilateral hyperkeratosis of nipple and areola associated with androgen insensitivity and oestrogen replacement therapy. PMID- 11730070 TI - Cognitive subprocesses and schizophrenia. A. Reaction-time decomposition. AB - OBJECTIVE: The aim of the study is to demonstrate that deficits of information processing in schizophrenic patients can be isolated with reaction-time (RT) decomposition paradigms. METHOD: Three types of visually presented tasks were applied: simple, disjunctive and choice RT-tasks. RT were split into movement latency and time necessary to execute movements. Comparisons of three samples of schizophrenic patients (295.3) with individually matched (age, sex. education and handedness) healthy controls are presented: Sample 1: 10 drug-naive first-onset patients, Sample 2: 10 neuroleptically treated first-onset patients, Sample 3: 10 neuroleptically treated chronically ill patients. RESULTS: Findings indicate that schizophrenia affects primarily subprocesses in which percepts are translated into appropriate actions (response-selection). Neuroleptic treatment improves processing at this stage but is accompanied by slowing of movement execution. CONCLUSION: Response-selection is selectively impaired in first-onset patients. This disturbance, which might be specific for schizophrenia, can be regarded as indication of a disconnection between frontal and posterior areas. PMID- 11730069 TI - A novel pharmacological concept in an animal model of psychosis. AB - OBJECTIVE: We have analysed pharmacologically induced perturbation of functional and structural neurogenesis in the prefrontal cortex (PFC) and hippocampus. METHOD: Juvenile gerbils received a single dose of methamphetamine (METH, 50 mg/kg, i.p.). In adults the following parameters were quantitatively investigated: prefrontal dopaminergic and GABAergic innervation densities (immunocytochemistry), morphogenesis of pyramidal cells (Golgi), dentate granule cell proliferation (BrdU-labelling), working memory and behavioural inhibition (delayed response, open-field). RESULT: A single challenge of METH continuously suppresses granule cell proliferation in adult gerbils and initiates rewiring of neuronal networks in the PFC which run concurrently with the development of severe deficits in PFC-related behaviours. CONCLUSION: It appears that a continuous remodelling of neuronal circuits is an inherent property of the brain, the biological significance of which seems to be to ascertain adaptive interaction between brain and environment. Learning more about drug-induced neuronal reorganization might be basic for understanding the genesis of psychotic conditions in the brain. This presentation is based both on own research and on a review of the literature. PMID- 11730071 TI - Cognitive subprocesses and schizophrenia. B. Maze tasks. AB - OBJECTIVE: In a companion paper we demonstrated that in reaction-time tasks the response selection stage is selectively disturbed in schizophrenia. The aim of the present study is the investigation of subprocesses, which are incorporated into repeatedly activated loops of cognitive processes and need additional information from working memory. METHOD: Maze tasks of varying complexity (with and without bifurcations) were presented to drug-naive and treated first-onset schizophrenic patients. The tasks had to be solved under easy and difficult motor conditions. Both patient samples were compared with healthy control groups matched by age, sex, education and experience with the experimental setting. RESULTS: In mazes with alternative routes patients exhibited the expected decrease of performance. However, untreated patients improved under difficult motor conditions. This improvement is smaller after neuroleptic treatment. CONCLUSION: Under difficult motor conditions attention is focused on the execution of movements and thus withdrawn from perceiving irrelevant response alternatives, i.e. the effects of disturbed response selection are reduced. PMID- 11730072 TI - Reaction-times and bioelectrical brain signals of drug-naive schizophrenic first onset patients in identification and classification tasks. AB - OBJECTIVE: The question of the present study is whether disturbances of response selection in schizophrenic patients are discernible only if overt motor-actions are required, or also if covert cognitive actions are necessary. METHOD: Visual identification (digits) and classification (dot-enumeration) tasks were presented to 18 drug-naive, first-onset schizophrenic patients and healthy controls. It is assumed that enumeration of more than three dots requires additional cognitive processes as buffering and re-focusing of attention. Reaction-times and 21 channel-EEG were measured. For eye-movement artefact-elimination a new non parametric regression approach was applied. RESULTS: Reaction-times revealed that in the patient group response selection is lengthened in both tasks. Perception of dot numbers is not affected. Bioelectrical data depicted a left-lateralization of posterior P100 and N 100 in the patient group as well as an enhanced fronto central P200. CONCLUSION: Whereas in reaction-times of patients only a disturbance of response selection is discernible, bioelectrical measurements also point to an altered organization of perceptive processes. PMID- 11730073 TI - Cognitive dysfunction in schizophrenia. PMID- 11730074 TI - Neuropsychological impairments in patients with schizophrenia: stability and prediction of outcome. AB - OBJECTIVE: To assess neuropsychological impairments among fairly young schizophrenic patients over time, and relations between impairment and symptoms, drug type and dose. METHOD: Clinical and neuropsychological data for 29 schizophrenic patients have been published earlier. Twenty of these patients were retested after 3 years using the same methods. RESULTS: The patients displayed stable impairments in most neuropsychological tests. Vocabulary and focused attention improved over time, whereas response slowness became more pronounced. Symptoms varied considerably over time. A specific pattern of neuropsychological impairments, but not measures of psychopathology, predicted clinical outcome. Patients were unable to judge their test performance. CONCLUSION: A few years after the first episode, our patients displayed a specific pattern of neuropsychological impairment, which predicted clinical outcome. One aspect of the impairment appears to be failure at self-monitoring. PMID- 11730075 TI - Schizophrenia--progressive and massive decline in response readiness by episodes. AB - OBJECTIVE: To relate the pattern of neuropsychological impairments among schizophrenic patients to case history data and disease characteristics in a cross-sectional study of unselected patients, and to integrate these data with two previous longitudinal studies of neuropsychological impairments among schizophrenic patients. METHOD: One hundred consecutive schizophrenic patients were studied with respect to clinical case history and current symptoms, medication and neuropsychological impairment using a comprehensive computerized test battery. RESULTS: The most salient finding was a marked slowing of response readiness, linearly related to the number of previous acute episodes. The resulting deficit was far beyond what has been obtained in any other group of subjects (average -6 SD for >five episode patients). The impairments in many of the other neuropsychological parameters could to some extent be explained with reference to response slowing, with one exception--verbal short-term memory. Adjustment for important confounding factors (age, duration of illness, medication) did not change the strong negative association between response readiness and number of previous episodes. CONCLUSION: These findings, together with findings of our two previous longitudinal studies and a recent replication, prompted us to suggest that each acute schizophrenic episode inflicts damage to a set of hypothetical structures, cognitive pattern generators. We assume that these structures translate intentions to logistic programs. When damaged, delays are introduced into executive functions and corollary discharge processes will run out of phase with intentions. This model implicates new ways of looking at the generative mechanisms of the illness, and on treatment strategies. PMID- 11730076 TI - Neuropsychological impairment in patients treated with depot neuroleptics: a longitudinal study. AB - OBJECTIVE: To assess neuropsychological impairments among chronic functional psychotic patients over time, and relations with symptoms, drug dose and side effects. METHOD: Thirty-four patients, representative of the most ill one-third of all patients with chronic functional psychoses known to the psychiatric services in a city catchment area, were assessed for clinical symptoms, drug side effects and by neuropsychological tests at study entry. They were then assessed repeatedly over 2 years. RESULTS: All patients were seriously impaired in the tests. The impairment was stable over time, in spite of substantial changes in the clinical state. The impairment was unrelated to drug and drug dose. Patients with prominent negative symptoms were most impaired and most unable to rate their performance in the tests. CONCLUSION: Patients with chronic functional psychoses do relapse often, also late in the course of the disease. Negative symptoms, marked impairments in simple neuropsychological tests and impaired self monitoring went together. PMID- 11730077 TI - Neuropsychological performance of psychotic patients in community care: results from the UK700 study. AB - OBJECTIVE: To compare cognitive performance in chronic schizophrenic and affective psychotic patients maintained in community care. METHOD: We studied a sample of community-based patients (n = 707) with chronic psychotic disorders. Neuropsychological assessment was completed using the National Adult Reading Test (NART) and the Trail Making Test (TMT). RESULTS: Affective psychotic patients had higher premorbid IQ than schizophrenic patients before adjustment for confounding factors (P=0.03); however, after adjustment for ethnic group and social class this became non-significant (P=0.19). There were no significant differences between groups on the TMT, parts A or B. CONCLUSION: Unlike studies suggesting that schizophrenic patients are more cognitively impaired than affective psychotic patients, our study suggests a degree of cognitive homogeneity between those patients who develop a chronic illness. Measures of premorbid IQ suggest that this cognitive homogeneity exists prior to the onset of illness. PMID- 11730078 TI - Ruth L. Kirschstein, MD. PMID- 11730079 TI - Financial incentive for young clinical researchers: loan repayment programs, public and private. PMID- 11730080 TI - Stem cell research proceeds, registry created. PMID- 11730081 TI - Maryland court of appeals decision threatens research at Kennedy Krieger Institute. PMID- 11730082 TI - Implementation of a data and safety monitoring program in a general clinical research center. PMID- 11730083 TI - Clinician scientist training program: a proposal for training medical students in clinical research. AB - There is national alarm about a decline in the number of clinician scientists. Most of the proposed solutions have focused on housestaff and junior faculty. We propose a new national program for training medical students in clinical research. This program, coined "Clinician Scientist Training Program" (CSTP), would consist of a combined degree program in medicine (MD) and clinical research (eg, masters in translational research or masters in clinical epidemiology). Students could enroll in the program at any stage during medical school. After 3 years of medical school, students would spend at least 2 years in a combined didactic and mentored clinical research training program and then complete medical school. Students could elect to pursue more prolonged clinical research training toward a combined PhD and MD. The CSTP is designed to meet six critical challenges: 1) engage students early in clinical research training; 2) provide a didactic clinical research curriculum; 3) expose students to several years of mentored clinical research training; 4) promote debt prevention by providing tuition payments during medical education and a stipend during clinical research training; 5) facilitate prolonged exposure to a community of peers and mentors in a program with national and institutional identity and respect; and 6) permit enrollment in the program as students enter medical school or at any stage during medical school. If the success of the Medical Scientist Training Program in training medical students in basic research is a guide, the CSTP could become a linchpin for training future generations of clinician scientists. PMID- 11730084 TI - High-Resolution magnetic resonance imaging of normal and atherosclerotic human coronary arteries ex vivo: discrimination of plaque tissue components. AB - BACKGROUND: Noninvasive detection of plaque lipid and calcium in human atherosclerosis may have clinical utility, because the presence of each may be associated with increased risk of plaque disruption. Magnetic resonance imaging (MRI) has the potential to detect both plaque lipid and calcium. However, no previous studies have: 1) used an MR coil with sufficient resolution to image the components of human coronary arteries, 2) evaluated the utility of a combination of different MR contrast weightings in discriminating plaque components in human coronary arteries, or 3) used sensitive and specific histological stains for lipid and calcium to determine their MR image characteristics in human atherosclerosis. METHODS: Using a custom-made surface coil on a whole-body, 1.5T MRI scanner, high resolution MR images were obtained from 22 nonatherosclerotic and atherosclerotic human coronary artery segments and then compared with histological sections stained for neutral lipid, calcium, and ribrous and cellular components. RESULTS: With a multicontrast protocol using T1-, proton density-, and T2-weighted images, statistically significant differences were found among MR image contrast values for regions identified by histological stains as containing lipid only, calcium only, mixed lipid and calcium, or fibrous tissue. All four of these histologically defined region types could be differentiated from one another by a multicontrast MRI protocol. Of the 22 segments, 10 (45%) contained areas with combined plaque lipid and calcium; calcium would not have been recognized histologically in these regions without the use of a specific calcium stain. CONCLUSIONS: These results demonstrate that multicontrast MRI can produce remarkably high-resolution images and can discriminate between clinically relevant components of the atherosclerotic vessel wall. PMID- 11730085 TI - Acute and chronic hemodynamic effects of enhanced external counterpulsation in patients with angina pectoris. AB - BACKGROUND: Enhanced external counterpulsation (EECP) is an effective noninvasive treatment for patients with angina pectoris. However, the hemodynamic effects of EECP are still unknown and have been theorized to simulate the clinical use of the intra-aortic balloon pump, enhancing cardiac output, stroke volume, and retrograde aortic diastolic flow. METHODS: Twelve hemodynamic parameters were measured, using the BioZ System (CardioDynamics International Corporation, San Diego, Calif) after 1 hour (n=22) and after 35 hours (n=16) of EECP treatment compared with baseline. The BioZ System noninvasively measures hemodynamic parameters using the thoracic electrical bioimpedance method. RESULTS: One hour of EECP treatment revealed a significant decrease in cardiac output, stroke volume, contractility, afterload, preload, and myocardial energy production, but systolic time ratios and thoracic fluid content were unchanged. However, after 35 hours of therapy, stroke volume (P < or = 0.05), index of contractility (P < or = 0.05), and thoracic fluid content (P < or = 0.01) were decreased. CONCLUSIONS: The acute reduction in contractility, preload, and concurrent bradycardia may have favorable hemodynamic effects for patients with angina pectoris. Further studies are needed to elucidate the mechanism of EECP therapy and its efficacy for patients with angina pectoris. PMID- 11730086 TI - Relationship of early carotid artery disease with lipoprotein (a), apolipoprotein B, and fibrinogen in asymptomatic essential hypertensive patients and normotensive subjects. AB - BACKGROUND: We investigated the relationships between plasma lipids and lipoprotein fractions and carotid artery lesions (CAL) in 177 cerebro-vascularly asymptomatic subjects, of whom 107 were primary hypertensive patients and 70 normotensive controls. METHODS: The prevalence and severity of CAL, as assessed by calculating a score of severity (score of CAL) and the maximal stenosis of both sides, as well as the intimal-medial thickness (IMT) were evaluated with a high-resolution echo-Doppler technique. We measured total serum cholesterol, triglycerides, low-density lipoprotein-cholesterol, high-density lipoprotein cholesterol, lipoprotein (a) [Lp(a)], Apo (apolipoprotein)AI, ApoAII, ApoB, and fibrinogen. RESULTS: Both the prevalence (59.4% vs 26.2%) and severity of sex- and age-adjusted and unadjusted CAL and IMT were significantly higher in hypertensive patients than in controls. Regression analysis showed different predictors of IMT and maximal stenosis. The variables that remained in the model were age, mean blood pressure (BP), and smoking for IMT; pulse pressure, known duration of hypertension (HT), fibrinogen, and ApoB for the score of CAL; and the last four variables along with age and mean BP for maximal stenosis. Furthermore, we identified a link between the atherogenic lipoprotein fractions Lp(a) and ApoB, fibrinogen and early carotid artery atherosclerotic changes. CONCLUSIONS: The different correlates of IMT, CAL, and maximal degree of stenosis suggest that they reflect different events occurring in the arterial wall in response to aging, HT, and other risk factors, rather than simply different stages of the same atherosclerotic process. PMID- 11730087 TI - Calcitonin gene-related peptide expression in sepsis: postulation of microbial infection-specific response elements within the calcitonin I gene promoter. AB - BACKGROUND: Recently, we reported an unexpected ubiquitous expression of calcitonin (CT)-mRNA in a hamster peritonitis model of sepsis. Using this animal model,we undertook a study to further investigate the pattern of expression of the calcitonin I (CALC-I) gene and CT gene-related peptide (CGRP)-mRNA in sepsis. METHODS: Live Escherichia coli impregnated in agar pellets were implanted in the peritoneal cavities of hamsters. Twelve hours after sepsis induction, the septic and healthy control animals were sacrificed and tissues and peritoneal macrophages were collected. CGRP-mRNA content was evaluated by reverse transcription polymerase chain reaction (RT-PCR), quantitated by the Taq-Man technique, and compared with the mRNA expression of CT, tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6). The 5' untranslated regions of the mRNA and potential alternative splicing sites were identified by 5' rapid amplification of cDNA ends. RESULTS: We found a tissue-wide, ubiquitous and uniform expression of CGRP-mRNA in all septic tissues examined. CGRP-mRNA was detectable by RT-PCR in various extraneuronal and extrathyroidal septic tissues, but not in healthy control tissues. As found for CT-mRNA in our earlier studies, CGRP-mRNA seemed to be more specifically up-regulated as compared with other classical cytokines (ie, II-6 and TNF-alpha). Importantly, the 5' untranslated sequence in control and septic thyroid was similar to the sequence obtained from septic spleen. CONCLUSIONS: We postulate the presence of microbial infection specific response elements in the CALC-I gene promotor, which, upon a specific stimulus, override the tissue-selective expression pattern. This new form of endocrine plasticity may be of importance in the response to systemic inflammation. PMID- 11730088 TI - Fenamates stimulate BKCa channel osteoblast-like MG-63 cells activity in the human. AB - BACKGROUND: The fenamates, a family of nonsteroidal anti-inflammatory drugs that are derivatives of N-phenylanthranilic acid, are the inhibitors of cyclo oxygenase. The ionic mechanism of actions of these compounds in osteoblasts is not well understood. METHODS: The effects of the fenamates on ionic currents were investigated in a human osteoblast-like cell line (MG-63) with the aid of the whole-cell and inside-out configurations of the patch-clamp technique. RESULTS: In MG-63 cells, niflumic acid and meclofenamic acid increased K+ outward currents (IK). The niflumic acid-stimulated IK was reversed by subsequent application of iberiotoxin or paxilline, yet not by that of glibenclamide or apamin. In the inside-out configuration, niflumic acid (30 micromol/L) added to the bath did not modify single-channel conductance but increased the activity of large-conductance Ca2+-activated K+ (BKCa) channels. The EC50 values for niflumic acid- and meclofenamic acid-induced channel activity were 22 and 24 micromol/L, respectively. Niflumic acid (30 micromol/L) and meclofenamic acid (30 micromol/L) shifted the activation curve of BKCa channels to less positive membrane potentials. Membrane stretch potentiated niflumic acid-stimulated channel activity. The rank order of potency for the activation of BKCa channels in these cells was niflumic acid = meclofenamic acid > tolfenamic acid > flufenamic acid > nimesulide. Evans blue and nordihydroguaiaretic acid increased channel activity; however, indomethacin, piroxicam, and NS-398 had no effect on it. CONCLUSIONS: The fenamates can stimulate BKCa channel activity in a manner that seems to be independent of the action of these drugs on the prostaglandin pathway. The activation of the BKCa channel may hyperpolarize the osteoblast, thereby modulating osteoblastic function. PMID- 11730089 TI - Transesophageal dobutamine stress echocardiography with tissue Doppler imaging for detection and assessment of coronary artery disease. AB - BACKGROUND: Transesophageal dobutamine stress echocardiography (T-DSE) has been shown to be a sensitive and specific technique for the detection of myocardial ischemia. A major limitation of echocardiographic study interpretation, however, is the subjective visual analysis of endocardial motion and wall thickening, which is only semiquantitative. METHODS: To analyze whether T-DSE with the use or tissue Doppler imaging (TDI) during graded dobutamine infusion may be useful to detect and quantify stress-induced myocardial ischemia by changes in myocardial velocities, 70 patients undergoing coronary arteriography were studied with T-DSE and TDI. Midesophageal and transgastric short- and long-axis images were obtained at each level of dobutamine infusion. T-DSE was successful in 67 patients (96%). Baseline resting pulsed and color peak systolic (S) and early diastolic (E) velocities of the anterior, septal, lateral, and inferior walls were examined. RESULTS: Pulsed and color TDI correlated well at rest and after stress. Fifteen patients had a normal response to dobutamine, and 52 patients had inducible ischemia by two-dimensional criteria. In the normal group, there was a significant dose-dependent increase in S and E velocities. Compared with those in the normal group, patients with coronary artery disease (CAD) had lower resting S and E velocities and blunted S wave increase or E wave decrease during DSE. CONCLUSIONS: T-DSE with TDI is a feasible and accurate test for the quantitative assessment of patients with CAD who have impaired augmentation of systolic and diastolic myocardial velocities during dobutamine infusion. PMID- 11730090 TI - Symposium summary, Experimental Biology 2001. PMID- 11730091 TI - Clinical results after islet transplantation. PMID- 11730092 TI - Insights into alloislets from successful autoislet transplantation in patients with chronic pancreatitis. PMID- 11730093 TI - Heme oxygenase-1 protects pancreatic beta cells from apoptosis caused by various stimuli. AB - BACKGROUND: Several problems can occur after allogeneic islet transplantation: primary nonfunction, rejection, and the recurrence of autoimmune disease, which involve attack by the recipient's cytokines, T cells, natural killer cells, and monocytes on the donor's beta cells, which leads to beta-cell destruction. Recent studies have revealed that loss of transplanted islets is caused mainly by apoptosis. Heme oxygenase-1 (HO-1) is one of the antiapoptotic genes up-regulated under stress conditions. The aim of this work was to investigate any mechanisms of HO-1-mediated protection of beta cells from apoptosis. METHODS: Apoptosis was assessed by comparison of viable transfected cells with and without apoptotic stimuli, and with and without HO-1 overexpression. Activation and function of p38 mitogen-activated protein kinase were determined using the specific inhibitor SB203580. RESULTS: We have shown that HO-1 mediates antiapoptotic effects in beta cells. The percentage of apoptotic cells after stimulation with tumor necrosis factor a decreased from 75% without HO-1 to 5% when HO-1 was overexpressed. Our data indicate that HO-1 acts as a signal terminator of tumor necrosis factor alpha-induced apoptosis by modulation of the p38 mitogen-activated protein kinase pathway. CONCLUSIONS: Profound cell stress that occurs in islets after transplantation, as well as at the onset of diabetes, results in beta-cell loss through apoptosis. Protection of beta cells by HO-1 improves their survival in vitro after various proapoptotic stimuli, suggesting that HO-1 suppresses one or several signaling pathways leading to apoptosis. We hypothesize that our in vitro findings can be extrapolated to the in vivo situation, and we propose that expression of HO-1 in islets may illuminate a valuable new approach to improving diabetes treatment. PMID- 11730094 TI - Prevention of morphological changes in alginate microcapsules for islet xenotransplantation. AB - BACKGROUND: Alginate microcapsule swelling, which occurs as a result of increased hydrophilicity owing to the Ca++ that remains after rapid chelation of the inner alginate core, is a problem in encapsulation. We have previously shown that exchange of the residual divalent Ca++ with the monovalent Na+ through the use of 6 mmol/L Na2SO4 decreases swelling in chelated alginate-polylysine-alginate microcapsules, and this process enhances their durability. The purpose of the present study was to examine the morphology of Na2SO4-treated microcapsules in long-term incubation with the use of serum-supplemented culture medium. METHODS: Spherical beads of purified alginate (3%) that were gelled with 1.1% CaCl2 were first coated with polylysine, and then with 0.24% alginate. After rapid chelation of the inner alginate core with 55 mmol/L sodium citrate, the capsules were either incubated for 30 minutes in 6 mmol/L Na2SO4 or left untreated (control). Each group of capsules was then placed in a flask containing Ham's culture medium supplemented with 20% porcine serum and incubated at 37 degrees C. RESULTS: The diameters of Na2SO4-treated capsules only increased modestly from a mean +/- SD of 635 +/- 22.08 to 684.53 +/- 17.86 microm (P<0.0001) by day 7, with no further increases thereafter. In contrast, control capsules showed a steady increase in their mean diameters, which changed from 639.55 +/- 21.44 to 735.48 +/- 108.85 microm (P < 0.0001) by day 66. In addition, whereas treated capsules remained spherical, control capsules showed progressive polymorphism. CONCLUSION: We have developed a new method of making more durable and stable microcapsules that can be used for islet cell xenotransplantation. PMID- 11730095 TI - Porcine islets as an alternative to human islets for transplantation. PMID- 11730096 TI - Methods for volume measurement in 3D images. PMID- 11730097 TI - Viscoelasticity of the vessel wall: the role of collagen and elastic fibers. AB - The aortic wall contains collagen fibrils, smooth muscle cells, and elastic fibers as the primary load-bearing components. It is well known that the collagen fibrils bear loads in the circumferential direction, whereas elastic fibers provide longitudinal as well as circumferential support. Stiffening of the vessel wall is associated with loss of elastic tissue and increases in the collagen content: however, little is known about the mechanism of vessel wall stiffening with age. The purpose of this review is to attempt to relate structural changes that occur to the collagen and elastic fibers to changes in the viscoelastic behavior that are associated with aging. Analysis of the viscoelastic mechanical properties of collagen fibrils from tendon, skin, and aortic wall suggest that the collagen fibrils of aortic wall are different than those of other tissues. The elastic spring constant of the collacen fibrils in vessel walls is significantly less than that found in tendon, suggesting that the presence of type III collagen in aortic wall increases the flexibility of the collagen fibrils. Furthermore, it is hypothesized that changes in the interface between collagen fibrils, elastic fibers, and smooth muscle during aging and in connective tissue disorders leads to changes in the viscoelasticity of the vessel wall. PMID- 11730098 TI - Computational modeling of ligament mechanics. AB - This article provides a critical review of past and current techniques for the computational modeling of ligaments and tendons. A brief overview of relevant concepts from the fields of continuum mechanics and finite element analysis is provided. The structure and function of ligaments and tendons are reviewed in detail, with emphasis on the relationship of microstructural tissue features to the continuum mechanical hehavior. Experimental techniques for the material characterization of biological soft tissues are discussed. Past and current efforts related to the constitutive modeling of ligaments and tendons are classified by the particular technique and dimensionality. Applications of one dimensional and three-dimensional constitutive models in the representation of the mechanical behavior of joints are presented. Future research directions are identified. PMID- 11730099 TI - Equicluster water model. AB - A new cluster model of water is presented. Using this model, we estimated basic thermodynamic parameters of water. A new mechanism of dielectric polarization is proposed. It explains a small spread in the dielectric relaxation time of water. PMID- 11730100 TI - Simulation of broadband dielectric spectra of water and aqueous electrolytic solutions in terms of the intermolecular potential einartig representation. PMID- 11730101 TI - Numerical simulation of light propagation and scattering in turbid biological media. AB - This article considers the processes of light propagation and scattering in biological tissues. The results obtained made it possible to estimate basic signal parameters and their dependence on various optical parameters with respect to the laser Doppler flowmeter and laser Doppler microscope. We also developed a new method to determine the indicatrix asymmetry of single and multiple light scattering by a suspension of oriented spheroidal particles that simulated erythrocytes in a shear flow. It was found that the angular dependence of the asymmetry index provides infonnation on the shape and orientation of particles. In addition, we obtained single scattering indicatrices, which may improve the accuracy of computer simulation of light scattering by blood. PMID- 11730102 TI - Key biological structures of the human being's informational image. AB - This article deals with the corporeal and informational images of human beings. These images can be represented by key biological structures, such as biocells, nerve fibers, and acupuncture points. These structures are shown to transmit and process informational signals by means of electromagnetic (field) waves. An experimental setup is designed to study biological structures in the millimeter wave range. PMID- 11730103 TI - Effect of low-intensity infrared and millimeter radiation on higher plants' biopotentials. AB - This article studies the effect of local low-intensity electromagnetic radiation on the bioelectric responses of plants. In our investigation, we used thirty three wavelengths in the visible and infrared spectrurm regions as well as three wavelengths in the millimeter spectrum region. As a result, we obtained the bioelectric responses of plants to electromagnetic radiation not only in the absorption region of cellular pigments (such as chlorophyll, flavin, and phytochrome) but also in the absorption region of water molecules. PMID- 11730104 TI - Electromagnetic perception and individual features of human beings. AB - An investigation was made of the individual reactions of human subjects exposed to electromagnetic fields. We performed the study on 86 volunteers separated into two groups. The first group was exposed to the electromagnetic field of infralow frequencies, whereas the second group was exposed to the electromagnetic field of extremely high frequencies. We found that the electromagnetic perception of human beings correlated with their individual features, such as EEG parameters, the critical frequency of flash merging, and the electric current sensitivity. Human subjects who had a high-quality perception of electromagnetic waves showed an optimal balance of cerebral processes, an excellent functional state of the central nervous system, and a good decision criterion. PMID- 11730105 TI - Constant electric field of human beings and its role in medicine. AB - An investigation was made of fundamental biophysical principles of the intrinsic electric system functioning in different conditions. This system was shown to govern the vital functions of separate cells and of the entire live organism. We developed a method enabling the correction of an affected intrinisic electric system. This method is based on the application of an external electric current. PMID- 11730106 TI - Comparative analysis of methods for classifying the cardiovascular system's states under stress. AB - A comparative analysis is made of various methods for processing electrocardiograms and RR-interval sequences. This analysis was carried out by using standard nonlinear-dynamics algorithms and methods. Apart from that, we assessed the expediency of using a number of characteristics to classify the cardiovascular system's state under stress. PMID- 11730107 TI - Application of millimeter waves in treating chronic prostatitis patients. AB - A comparative analysis was made of various EHF therapy treatments applied in chronic prostatitis patients. The therapeutic efficiency of these treatments was assessed on a large number of patients. Besides general clinical examination, we employed ultrasonography, seminal analysis, local immununoassay, and the Voll diagnosis technique. It was found that, when millimeter waves were tuned to bioresonance, rectal treatment provided the highest clinical efficiency. PMID- 11730108 TI - Laser radiation control in photodynamic therapy of tumors. AB - This article presents the ideology and basic principles of designing diagnostic and therapeutic medical systems for treating malignant tumors by means of photosensitizers. The designed system is based on optoelectronic devices, which realize selective laser tumor destruction during photodynamic therapy. A distinguishing feature of this system is that laser-beam formation is performed in real time. This enables one to follow changes in the tumor shape during the treatment. As a result of this study, a photodynamic technique for selective phototreatment was developed and tested. Moreover, we detemined light doses for such treatment. PMID- 11730109 TI - Design and application of low-frequency ultrasound and its combination with laser radiation in surgery and therapy. AB - Further development of combined laser-ultrasonic medical technologies is presented. An analysis is made of the main effects produced by low-frequency ultrasound on biological tissues. These effects include cavitation, acoustic streamings, acoustic pressure, and mechanical influence. Possible application of low-frequency ultrasound is considered in regard to tumor therapy, bactericidal treatment of infectious tissues, and spray treatment of head and neck wounds. Some limitations of ultrasound application in medicine are discussed, and the feasibility of combining laser radiation with ultrasound is studied. A number of new laser-ultrasonic combinations are put forward and developed. They embrace the combination of photodynamic therapy with ultrasonic tumor therapy (sonophotodynamic therapy), laser-ultrasonic wound treatment (spray treatment), and laser-ultrasonic drug delivery (photophonophoresis). Some of the above mentioned technologies were tested experimentally, and the results obtained are considered. PMID- 11730110 TI - Investigation and development of ultrasonic vibrodrives for traumatology. AB - An investigation was made of the dynamics of ultrasonic vibrodrills with a common source of rotational and ultrasonic reciprocal motions. The source was constructed on the basis of an electroacoustic transducer of longitudinal vibrations. The dependences of power, rotation speed, and flutter rates were plotted for various pushers as a function of biological tissue drag torque. The results obtained allowed us to determine the optimal parameters of basic ultrasonic modes used in treating biological tissues. Apart from that, ultrasonic vibrodrill pilot models, which are new in principle, are presented. They were designed to be applied in traumatology. A distinguishing feature of these vibrodrills is that an electroacoustic transducer of longitudinal vibrations is a source of two motion types: rotational and ultrasonic reciprocal. PMID- 11730111 TI - Investigation of blood flow microcirculation by diffusing wave spectroscopy. AB - This article presents a technique and a setup for noninvasive in vivo studies of cutaneous microcirculation. The technique is based on the analysis of intensity fluctuations of scattered coherent light. The analysis is made in terms of diffusing wave spectroscopy. The developed technique can be used in clinical practice to determine the motion pattern of scattering particles (such as red blood cells) in randomly inhomogeneous media (such as biological tissues). The average size of scattering particles can be comparable to the wavelength of scattered radiation. PMID- 11730112 TI - Laser doppler flowmetry as a method for evaluating the microwave radiation effect on cutaneous microcirculation. AB - It is demonstrated that laser Doppler flowmetry can be used to evaluate the effect of low-intensity microwave radiation on cutaneous microcirculation. PMID- 11730113 TI - Rayleigh light scattering and electron paramagnetic resonance methods for ecological monitoring and medical diagnostics. AB - Two physical methods, such as Rayleigh light scattering and electron paramagnetic resonance (EPR), are proposed for detecting toxic heavy metals in aqueous media. These methods can find application in ecological monitoring and medical diagnostics. The advantages of these methods over existing ones are considered, and the threshold values of heavy metal concentrations detected by various physical methods are compared. PMID- 11730114 TI - Incidence and predictors of coronary heart disease among older African Americans- the Cardiovascular Health Study. AB - Although coronary heart disease (CHD) is the leading cause of death and morbidity in older African Americans, relatively little is known about the incidence and predictors of CHD in this population. This study was undertaken to determine the incidence and predictors of CHD in African-American men and women aged 65 years and older. The participants in this study included a total of 924 African American men and women aged 65 years of age and older who participated in the Cardiovascular Health Study (CHS). The overall CHD incidence was 26.6 per 1,000 person-years of risk. Rates were higher in men than women (35.3 vs. 21.6) and in those 75 years or older than in those less than 75 years (31.3 vs. 24.5). In multivariate analysis, factors associated with higher risk of incident disease were male gender [relative risk (RR) = 1.8, 95% confidence interval (CI) = 1.1, 2.7], diabetes mellitus (RR = 1.9, 95% CI = 1.2, 2.9), total cholesterol (RR for 40 mg/dL increment = 1.3, 95% CI = 1.0, 1.5), and low (i.e., <0.9) ankle-arm index (RR = 2.1, 95% CI = 1.3, 3.4) after adjusting for age. Within this cohort of older African Americans, male gender, diabetes mellitus, total cholesterol, and low ankle-arm index and were independently predictive of incident events. These results suggest that the ankle-arm index, a measure of advanced atherosclerosis, should be further evaluated for its efficacy in identifying older African Americans at risk for incident clinical events. PMID- 11730115 TI - Assessing cardiovascular disease risk in women: a cultural approach. AB - BACKGROUND: Cardiovascular disease among American women is affected by a number of high-risk lifestyle factors, but little is known about the perceptions of high risk behavior among women in an inner-city population. The two purposes of this study were to identify the perceptions of an inner-city, predominantly African American community as they pertain to a high-risk lifestyle for cardiovascular disease as well as to develop a culturally sensitive survey instrument for women. METHODS: There were two components to the study. In the first, four focus groups were conducted to obtain qualitative data on women's attitudes and lifestyles regarding cardiovascular disease risk. In the second, focus group data were used to construct a survey on women's attitudes and lifestyles regarding cardiovascular disease risk that was modified using a fifth focus group and then pilot-tested with a sample of 27 women. RESULTS: Focus group and pilot-testing data suggest interesting differences between the behaviors and perceptions of inner-city women and the general population. OBESITY: Obesity was more loosely defined by this community than by guidelines based on standard height and weight measures. Being heavy was not necessarily equated with being fat and was felt at least partially to reflect muscle tone and muscle mass. STRESS: It was volunteered almost unanimously as a distinct risk factor for cardiovascular disease among women, although it rarely is listed on risk factor questionnaires. EXERCISE: Standard aerobic exercise participation was low, but participation in daily physical activity such as casual walking and housework was high. CONCLUSIONS: Health care providers, in attempting to reduce a patient's risk for cardiovascular disease, should be aware of the cultural and socioeconomic factors that might influence that patient's perceptions of cardiovascular disease risk. These perceptions should shape a provider's approach to lifestyle modification advice. PMID- 11730116 TI - Evaluation of a cervical cytology device (cell-sweep) based on comparison to colposcopic findings. AB - A new cytology sampling device, the CellSweep, identifies squamous intraepithelial lesions with a sensitivity of 75%. The purpose of this study was to evaluate the accuracy of cervical cytology using a new sampling device combining an endocervical brush and ectocervical spatula into one unit (CellSweep, patented by R. Mohajer, Troy, Michigan). From April 1995 to July 1995, 71 patients referred to the Allegheny University Hospitals Colposcopy Clinic had cervical cytology obtained with the CellSweep and underwent colposcopic evaluation of the cervix. The ability of the CellSweep to detect an abnormality confirmed by colposcopic evaluation was studied. Colposcopically directed ectocervical biopsies were obtained only in patients with identifiable lesions (n = 32). No random biopsies were obtained. The cytology smear was unsatisfactory for interpretation in one case. The remaining 70 Papanicolaou smears were read as normal in 17 (24%) cases and atypical squamous cells in 19 (27%). A squamous intraepithelial lesion (SIL) was detected in 34 (48%) smears. The colposcopic evaluation was normal in 50 patients who had satisfactory Papanicolaou smears, whereas SIL was detected in 20 cases. In 31 patients, SIL was not present in either colposcopy or cytology. In this preliminary study, the CellSweep identified SIL with a sensitivity of 75% and a specificity of 62%. The CellSweep, which combines an endocervical brush and an ectocervical spatula into a single unit, seems to be an acceptable device for obtaining cervical cells for cytologic screening. PMID- 11730117 TI - Laparoscopic intestinal injury: a review and case presentation. AB - BACKGROUND: The incidence of laparoscopic primary trocar small-intestine injury is unknown. The case presented here differed from a typical clinical course in that only excessive periumbilical fluid leak was present postoperatively. Neither classic symptoms nor signs were present to justify laparoscopic trocar small bowel perforations. CASE: A 42-year-old woman (G2 P1011, height 5'4", weight 132 lb) underwent elective, diagnostic, and operative laparoscopy with a lysis of extensive abdominal-pelvic adhesions for chronic pelvic pain. Preoperatively, the patient was classified as being at increased risk for intestinal laceration or perforation at the time of Veress needle or primary trocar insertion due to her surgical history. For this reason, mechanical bowel preparation with GoLYTELY was carried out. No intraoperative complications were noticed. After surgery, external, excessive fluid leak from the periumbilical incision only was observed (the three 5-mm incisions, in the lower part of the abdomen, were dry). Initially, this event was interpreted as residual irrigation fluid leakage. The patient was closely monitored for bowel injury, and neither medical condition nor laboratory tests changed from base within the initial 48 h, although excessive fluid drainage from periumbilical area was persistent. Enough time elapsed from laparoscopic surgery for CO2 and irrigation-fluid absorption; therefore, additional studies were ordered (an abdominal upright x-ray was inconclusive for viscous perforation and gastrointestinal x-ray with a water-soluble contrast medium documented small-intestine perforation). Exploratory laparotomy with partial bowel resection was executed. Postoperative clinical course was uneventful, and no long-term sequel was observed. CONCLUSIONS: 1) Persistent excessive external fluid leak from the periumbilical area after laparoscopic surgery with no drainage from other incisional sides may suggest small-bowel injury. 2) latrogenic, internal-external canalization between the small intestine and the skin masked clinical symptoms and signs of small-intestinal injury. 3) Lack of classic symptoms, signs, or changes in pertinent laboratory data did not rule out small-bowel perforation. PMID- 11730118 TI - Outcomes of training pediatricians to serve the underserved: social pediatrics. AB - The Residency Training Program in Social Pediatrics (RPSP) was established in 1970 as part of the Residency Training Program in Social Medicine administered by Montefiore Medical Center in the South Bronx of New York City in response to local need for physicians to practice in underserved populations in the inner city setting. We report on an analysis of the first 25 years of the RPSP, based on periodic surveys of all program graduates and demographic data. We conclude that our Social Pediatrics training program has been clearly successful in meeting its stated goal of producing physicians who will make a lifetime commitment to the practice of medicine in underserved communities. PMID- 11730119 TI - AIDS/HIV pandemic: the hidden agenda. PMID- 11730120 TI - After-hours telephone calls by parents at an inner-city community health center. AB - After-hours telephone consultation is a well-established component of the practice of pediatric medicine. We describe an assessment of the propriety of after-hours telephone calls made by parents at an inner-city pediatric clinic. We conclude that an inner-city pediatric clinic experienced neither more frequent nor less urgent calls in comparison to the data published for private practice suburban settings. PMID- 11730121 TI - Attitudes of primary care physicians toward cancer-prevention trials: a focus group analysis. AB - PURPOSE: Recruitment of low-income and minority women to cancer-prevention trials requires a joint effort from specialists and primary care providers. We sought to assess primary care providers' attitudes toward participating in cancer prevention trial recruitment. PROCEDURES: We conducted a focus group with seven Boston-based primary care providers serving low-income and minority women. Providers discussed knowledge, attitudes, and beliefs regarding their role in recruitment to prevention trials. FINDINGS: A qualitative analysis of the focus group transcript revealed nine categories. Three categories related specifically to the primary care physician: 1) the dual role physicians play as advocates for both patient and research; 2) threats to maintaining the primary care relationship; and 3) general philosophy toward prevention. An additional six categories could be subdivided as they apply to the primary care physician, the patient, and the community: 4) trust/commitment; 5) benefits of the research; 6) access to the research; 7) knowledge and recall of the research; 8) influences of media coverage about the research; and 9) cultural sensitivity. CONCLUSIONS: Investigators conducting cancer-prevention trials must address the concerns of primary care physicians to optimize recruitment of subjects- especially low income and minority women-into trials. PMID- 11730122 TI - Commencement address to Charles R. Drew University. PMID- 11730123 TI - Full-scale experiences with anaerobic treatment of industrial wastewater. AB - High rate anaerobic treatment for industrial wastewater was first applied on a commercial scale in the sugar industry in the mid 70's. Since that time the technology has developed into a standard method of wastewater treatment for a wide variety of industries. The technology is now functional in over 65 countries and a total of approximately 1,400 plants were build by the 16 leading vendors of such systems. These plants account for approximately 65% of the total number of anaerobic treatment plants for industrial applications, which is estimated to be ca. 2,000. From the analysis from a database consisting of 1,215 plants it appears that the UASB technology as originally developed in the Netherlands is the most predominant process. It is also seen that the higher loaded EGSB type systems are gradually replacing at least some of the UASB applications. PMID- 11730124 TI - Corrosion protection by anaerobiosis. AB - Biofilm-forming bacteria can protect mild (unalloyed) steel from corrosion. Mild steel coupons incubated with Rhodoccocus sp. strain C125 and Pseudomonas putida mt2 in an aerobic phosphate-buffered medium containing benzoate as carbon and energy source, underwent a surface reaction leading to the formation of a corrosion-inhibiting vivianite layer [Fe3(PO4)2]. Electrochemical potential (E) measurements allowed us to follow the buildup of the vivianite cover. The presence of sufficient metabolically active bacteria at the steel surface resulted in an E decrease to -510 mV, the potential of free iron, and a continuous release of ferrous iron. Part of the dissolved iron precipitated as vivianite in a compact layer of two to three microns in thickness. This layer prevented corrosion of mild steel for over two weeks, even in a highly corrosive medium. A concentration of 20 mM phosphate in the medium was found to be a prerequisite for the formation of the vivianite layer. PMID- 11730125 TI - Experiences and potential of anaerobic wastewater treatment in tropical regions. AB - In this article an overview is given on the experience with anaerobic treatment of domestic wastewater in the tropics, with emphasis on the situation in India. Some design criteria and their impact on the costs of UASB reactors are discussed. The operational results of a number of full-scale reactors are presented. The applicability of the UASB, in combination with different post treatment units to comply with a variety of effluent standards is compared with other systems. From the available data it is concluded that, if nitrogen removal is not required, a UASB system, followed by a polishing pond, is a relatively simple, affordable, and manageable wastewater treatment system. Trickling filters may follow UASB treatment if effluent standards require removal of TKN. The inclusion of UASB reactors in wastewater treatment schemes allowing for total nitrogen removal still needs further study. PMID- 11730126 TI - Potential of anaerobic digestion of complex waste(water). AB - Although they differ greatly in origin complex waste(water)s mainly consist of proteins, lipids, carbohydrates and sometimes lignin in addition. Hydrolysis is the first and generally rate-limiting step in the process of anaerobic digestion of particulate organic substrates. Hydrolysis of particulate polymers can be described by Surface Based Kinetics, but for use in practice the empirical first order relation is advised. Unlike the hydrolysis of protein and carbohydrate, lipid hydrolysis is hardly occurring in the absence of methanogenesis. The latter is probably a physical rather than a biological process and affects the choice for either a one- or a two-step (phase) anaerobic reactor. In the chain of collection and transport, complex wastes often become complex wastewaters simply because of dilution. Dilution not only changes the reactor technology to be applied but also complicates the post-treatment and possibilities for resource recovery. Combining concentrated with diluted waste streams will almost always end up in much more complicated treatment technologies. PMID- 11730127 TI - Anaerobes, aerobes and phototrophs. A winning team for wastewater management. AB - Current mainstream technologies for wastewater treatment, such as activated sludge and tertiary nutrient removal, are too costly to provide a satisfactory solution for the increasing wastewater problems in developing regions. Besides, these technologies do not allow for re-use of valuable energy and nutrients contained in the wastewater. In light of these limitations, it is important to reconsider the technology and strategic approaches in wastewater management. This paper introduces the "Cleaner Production" concept to sewage management, which combines two approaches: pollution prevention and re-use. Pollution prevention includes a shift towards low water use sanitation technology. The more concentrated wastewater, becomes more attractive for re-use oriented treatment schemes. A combination of anaerobic pre-treatment followed by macrophyte-covered stabilisation ponds is proposed for the effective recovery of energy and nutrients from sewage. By selecting optimal applications of the plant biomass and pond effluent, nutrients will end up as fish and crop protein. This contrasts favourably to tertiary nitrogen removal in activated sludge systems, which recycles ammonia through molecular nitrogen at the expense of energy and high costs. Macrophyte ponds are proposed as a key step in waste recycling, because these form the central unit of a recycling engine, driven by photosynthesis. The process is energy efficient, cost effective and applicable under a wide variety of rural and urban conditions. PMID- 11730128 TI - Anaerobic treatment: a key technology for a sustainable management of wastes in Europe. AB - Environmental regulations in the European Union, based on the concept of integrated prevention and control of pollution, are oriented towards the sustainability of the production processes, and this leads to better recovery of resources from raw materials, energy saving, etc. This philosophy introduces a new framework to Environmental Engineers, who have to make efforts concerning waste minimisation. During the last few decades technologies based on the anaerobic treatment of wastewaters and organic wastes have been applied successfully to a wide variety of problems. A case study on the impact of applying anaerobic technology to the treatment of wastewaters from the sugar industry in Spain is presented. Nowadays, processes based on anaerobic treatment appear to be an excellent option as the core of an integrated process for waste and wastewater treatment. PMID- 11730129 TI - Perspectives on anaerobic treatment in developing countries. AB - Developing countries occupy regions where the climate is warm most of the time. Even in sub-tropical areas, low temperatures do not persist for long periods. This is the main factor that makes the use of anaerobic technology applicable and less expensive, even for the treatment of low-strength industrial wastewaters and domestic sewage. Based mainly on papers presented at the "VI Latin-American Workshop and Seminar on Anaerobic Digestion" held in Recife, Brazil, in November 2000, this text approaches the perspectives of anaerobic treatment of wastewaters in developing countries. Emphasis is given to domestic sewage treatment and to the use of compact systems in which sequential batch reactors (SBR) or dissolvedair flotation (DAF) systems are applied for the post-treatment of anaerobic reactor effluents. Experiments on bench- and pilot-plants have indicated that these systems can achieve high performance in removing organic matter and nutrients during the treatment of domestic sewage at ambient temperatures. PMID- 11730130 TI - The development of anaerobic treatment and its future. AB - Anaerobic treatment is well over 100 years old. Its initial development was for the treatment of domestic wastewaters, using anaerobic filters and hybrid processes that are still of interest today. It then progressed in application to separate sludge digestion, then to treatment of dilute industrial wastewaters. Several processes have been developed that accomplish efficient treatment of wastewaters at short detention times. Major contributions to the broad application of anaerobic treatment and the better understanding of this process has come from efforts by G. Lettinga and his colleagues. They have emphasized its importance for meeting the need for sustainable development in the future. Greater efforts are now needed for broad application of anaerobic treatment for ridding the environment of unwanted organic materials by converting them into methane, a renewable energy source. PMID- 11730131 TI - Advances in high-rate anaerobic treatment: staging of reactor systems. AB - Anaerobic wastewater treatment (AnWT) is considered as the most cost-effective solution for organically polluted industrial waste streams. Particularly the development of high-rate systems, in which hydraulic retention times are uncoupled from solids retention times, has led to a world-wide acceptance of AnWT. In the last decade up to the present, the application potentials of AnWT are further explored. Research shows the feasibility of anaerobic reactors under extreme conditions, such as low and high temperatures. Also toxic and/or recalcitrant wastewaters, that were previously believed not to be suitable for anaerobic processes, are now effectively treated. The recent advances are made possible by adapting the conventional anaerobic high-rate concept to the more extreme conditions. Staged anaerobic reactor concepts show advantages under non optimal temperature conditions as well as during the treatment of chemical wastewater. In other situations, a staged anaerobic-aerobic approach is required for biodegradation of specific pollutants, e.g. the removal of dyes from textile processing wastewaters. The current paper illustrates the benefits of reactor staging and the yet un-exploited potentials of high-rate AnWT. PMID- 11730132 TI - Digestion and degradation, air for life. AB - Anaerobic degradation of dead biomass is a natural gasification process, an anaerobic crematorium producing a very useful end-product composed of methane and carbon dioxide, generally polluted with small amounts of some malodorous and quite toxic volatile S-compounds. It leads to the production of essential building elements for new life. This exciting field became my faith, vision, hope and expectation. This paper intends to present a reflection of more than three decades of research, teaching and advertisement in the field of sustainable environmental protection technologies, particularly of systems based on anaerobic digestion and the biological sulphur cycle. Considerable progress has been made during these decades worldwide, both in the basic understanding of the various processes and concepts, but also in the implementation of these systems, despite the fact that particularly the implementation frequently proceeded very laboriously. The difficulties certainly can no longer be attributed to technological limitations and/or insufficient understanding of the microbiology and chemistry only, but mainly to the frustrating social rigidity and short-term self-interest in all sectors of our society. By combining anaerobic processes with other microbiological degradation or transformation processes, like those based on the biological sulphur cycle, micro-aerobic and conventional aerobic and anoxic processes, ideal conditions can be created to valorise residues (wastes) from domestic, industrial and agricultural origin. It is simply not just "technology", but also a route to achieve more sustainability and justice in society. It is a fight against conservative establishments. Decomposition, disintegration disbandment, it also stands for deliverance and liberation, space and air for continuation of life. PMID- 11730133 TI - Future perspectives in bioreactor development. AB - The paper discusses conversion capacities of both anaerobic and aerobic wastewater treatment systems in relation to growth kinetics, hydrodynamics and biomass concentration. In the current modern anaerobic high-rate reactors the conversion potentials are optimally exploited. This is not yet true for aerobic systems since operation of aerobic systems under conditions of low biomass growth reduces the maximum applicable loading rates significantly. Both the concept of granulation and the introduction of fluidised bed systems have increased conversion capacities for both anaerobic and aerobic systems significantly. One of the latest development concerns the SBR with granular biomass. The grazing concept, in which ciliates convert aerobically grown dispersed cells, offers a possibility for significant improvement of aerobic systems. In the fields of psychrophilic and thermophilic anaerobic treatment, specific reactor development may contribute to further enhance volumetric conversion capacities. Due to reduced water usage, both COD and salt concentrations tend to increase for industrial effluents. As a consequence, there is a need for the development of anaerobic reactors retaining flocculant biomass. The membrane bioreactors offer a solution for certain niches in wastewater treatment. However the oxygen transfer economy is poor. There is a need for fundamental knowledge development to obtain a realistic image of this technology. PMID- 11730134 TI - Recalcitrance as a catalyst for new developments. AB - Humic substances are very recalcitrant in anaerobic environments as substrates, mostly due to their non-hydrolyzable high molecular weight structure, which is incompatible for cell uptake. The recalcitrant nature can be applied towards immobilizing pollutants into non-toxic humic substances. Certain anaerobic bioconversions such as demethylation of lignin and reduction of nitro- or azo groups can activate pollutants for humification processes upon subsequent exposure to air. Although humic substances provide no carbon or energy for microorganisms, the evidence presented here indicates that they are very active as terminal electron acceptors and redox mediators promoting the anaerobic degradation and biotransformation of priority pollutants. PMID- 11730135 TI - Temperature characteristics of methanogenic archaea and acetogenic bacteria isolated from cold environments. AB - In most terrestrial ecosystems of boreal and northern climate zones degradation of organic matter with methane production occurs at low temperature. Two psychrophilic methanogenic archaea and four acetogenic bacteria were described until now. Recently, we isolated 12 new strains of methanogenic archaea and 3 strains of acetogenic bacteria from different natural and man-made cold environments including tundra permafrost wet land, sediments of deep lakes, silt of sludge disposal pond, pig and cattle manure digested at 6 degrees C, and an anaerobic EGSB-reactor operated at 9 degrees C. The temperature characteristics of microorganisms isolated from cold environments are discussed. All isolates are able to grow below 10 degrees C, most of them grow at such low temperature as 1 degrees C. The upper temperature limit for most growing at low temperature acetogens is 30 degrees C, and the temperature optima is 20 degrees C and below. Most isolated methanogens have temperature optima around 25 degrees C, and upper temperature limits at 30-40 degrees C. Whether microorganisms able to grow at low temperature are classified as mesophiles, psychrophiles, or psychrotrophs (psychrotolerants) is an issue of this article. We propose to modify the basic temperature definition of anaerobic microorganisms growing at low temperature. PMID- 11730136 TI - The potential of anaerobic bacteria to degrade chlorinated compounds. AB - Chlorinated ethenes and chlorinated aromatics are often found as pollutants in sediments, groundwater, and wastewater. These compounds were long considered to be recalcitrant under anaerobic conditions. In the past years however, dechlorination of these compounds has been found to occur under anaerobic conditions at contaminated sites and in wastewater treatment systems. This dechlorination is mainly attributed to halo-respiring bacteria, which are able to couple this dechlorination to energy conservation via electron transport coupled phosphorylation. The dechlorinating activities of the halo-respiring bacteria seem to be confined to the dechlorination of chloroethenes and chlorinated aromatic compounds. In addition, methanogenic and acetogenic bacteria are also able to reduce the chlorinated ethenes via a-specific cometabolic pathways. Although these latter reactions may not be important in the remediation of contaminated sites, they may be of substantial influence in the start-up of remediation processes and in the application of granular sludge from UASB reactors. Specific halo-respiring bacteria may be used to increase the dechlorination activities via bioaugmentation in the case that the appropriate microorganisms are not present at the contaminated site or in the sludge. PMID- 11730137 TI - The biological sulfur cycle: novel opportunities for environmental biotechnology. AB - Although the study of sulfur cycle bacteria was already started around the 1890s by the famous microbiologists Winogradsky and Beijerinck, there are nowadays still many new discoveries to be made about the metabolic properties, phylogenetic position and ecological behaviour of bacteria that play a role in the biological sulfur cycle. The current interest of the scientific community in the biological sulfur cycle is very high, especially because of the many special organisms that have recently been discovered in the deep sea and other environments characterised by extreme conditions (such as high salt, low/high pH or temperature) and also in bioreactor environments. This paper highlights some of these new discoveries and relates them to environmental biotechnology. It is concluded that the micro-organisms from the sulfur cycle offer unique opportunities for sulfur pollution abatement and sulfur recovery. PMID- 11730138 TI - New developments in reactor and process technology for sulfate reduction. AB - Sulfate reduction has been regarded in the past as an unwanted process in anaerobic treatment of sulfate-rich wastewaters. Research efforts were primarily focused on H2S toxicity, competition between sulfidogenic and methanogenic microorganisms and suppression of sulfidogenesis. More recently, the potential sulfidogenesis for treating a wide range of wastestreams contaminated with oxidized sulfurous compounds and/or heavy metals was also appreciated. Heavy metals can be removed by the formation and subsequent precipitation of poorly soluble metal sulfides. Basically two approaches can be distinguished in wastewater treatment: passive treatment using low-cost technologies and active treatment in newly developed bioreactors. Both strategies are discussed. PMID- 11730139 TI - Methanol conversion in high-rate anaerobic reactors. AB - An overview on methanol conversion in high-rate anaerobic reactors is presented, with the focus on technological as well as microbiological aspects. The simple C1 compound methanol can be degraded anaerobically in a complex way, in which methanogens, sulfate reducing bacteria and homoacetogens interact cooperatively or competitively at substrate level. This interaction has large technological implications as it determines the final product of methanol mineralization, methane or carbon dioxide. The degradation route of methanol may be entirely different when environmental conditions change. Direct methanogenesis from methanol seems the predominant mineralization route under mesophilic conditions both in the absence and the presence of sulfate. Under thermophilic conditions methanol oxidation to carbon dioxide and hydrogen appears to play an important role. The UASB technology for mesophilic digestion of methanolic waste has presently reached full-scale maturity. The potential of methanol as feedstock for anaerobic processes is discussed. PMID- 11730140 TI - Combining the biological nitrogen and sulfur cycles in anaerobic conditions. AB - The biochemical processes involved in the anaerobic degradation of carbon, nitrogen and sulfur compounds can be represented by an oxidation-reduction or electron donor-acceptor scheme. The theoretic values of Gibbs free energy (deltaG0) calculated from thermodynamic data indicate the feasibility of the reactions. The interactions C-S and C-N are well known but there is a lack of information about the interaction N-S. The anaerobic transformation of nitrates using reduced sulfur compounds can be explained considering that nitrate acts as electron acceptor while reduced sulfur compounds are the electron donors. A new N S interaction in anaerobic conditions (ORP = -425 mV) has been experimentally observed when treating industrial wastewater rich in organic nitrogen and sulfate. The mass balances of the different nitrogenous and sulfur compounds in the liquid and gas phases clearly indicated an uncommon evolution. An important percentage of the nitrogen entering the reactor as TKN was removed from the liquid phase appearing as N2 in the gas phase. Simultaneously, only part of the sulfate initially present in the influent appeared as sulfide in the effluent or as hydrogen sulfide in the gas. These experimental observations may suggest a new anaerobic N-S biological interaction involving simultaneous anaerobic ammonium oxidation and sulfate reduction, ammonium being the electron donor and sulfate the electron acceptor. PMID- 11730142 TI - Solid waste digestors: process performance and practice for municipal solid waste digestion. AB - The most common types of anaerobic digesters for solid wastes have been compared based on biological and technical performance and reliability. Batch systems have the most simple designs and are the least expensive solid waste digesters. They have high potential for application in developing countries. Two-stage systems are the most complex and most expensive systems. Their greatest advantage lies in the equalisation of the organic loading rate in the first stage, allowing a more constant feeding rate of the methanogenic second stage. Two-stage systems with biomass accumulation devices in the second stage display a larger resistance toward toxicants and inhibiting substances such as ammonia. However, the large majority of industrial applications use one-stage systems and these are evenly split between "dry" systems (wastes are digested as received) and "wet" systems (wastes are slurried to about 12% total solids). Regarding biological performance, this study compares the different digester systems in terms of organic loading rates and biogas yields considering differences in input waste composition. As a whole, "dry" designs have proven reliable due to their higher biomass concentration, controlled feeding and spatial niches. Moreover, from a technical viewpoint the "dry" systems are more robust and flexible than "wet' systems. PMID- 11730141 TI - Industrial applications of new sulphur biotechnology. AB - The emission of sulphur compounds into the environment is undesirable because of their acidifying characteristics. The processing of sulphidic ores, oil refining and sulphuric acid production are major sources of SO2 emissions. Hydrogen sulphide is emitted into the environment as dissolved sulphide in wastewater or as H2S in natural gas, biogas, syngas or refinery gases. Waste streams containing sulphate are generated by many industries, including mining, metallurgical, pulp and paper and petrochemical industries. Applying process technologies that rely on the biological sulphur cycle can prevent environmental pollution. In nature sulphur compounds may cycle through a series of oxidation states (-2, 0, +2, +4, +6). Bacteria of a wide range of genera gain metabolic energy from either oxidising or reducing sulphur compounds. Paques B.V. develops and constructs reactor systems to remove sulphur compounds from aqueous and gaseous streams by utilising naturally occurring bacteria from the sulphur cycle. Due to the presence of sulphide, heavy metal removal is also achieved with very high removal efficiencies. Ten years of extensive laboratory and pilot plant research has, to date, resulted in the construction of over 30 full-scale installations. This paper presents key processes from the sulphur cycle and discusses recent developments about their application in industry. PMID- 11730144 TI - Is thrombophilia a factor in the development of hemiplegic cerebral palsy? AB - A population-based study of children with hemiplegic cerebral palsy (CP) was performed to investigate whether thrombophilic tendencies are implicated in the aetiology of the condition. Thirty-eight children (23 males, 15 females; mean age 8.7 years, SD 4.1 years) with hemiplegic CP were ascertained. Twenty-seven children(18 males, nine females; mean age 8.4 years, SD 4.3) gave consent for inclusion. The non-study group comprised five males and six females; mean age 9.4 years, SD 4.1. In six children, seven thrombophilic 'abnormalities' were identified. Five of these abnormalities were of an equivocal nature and probably did not represent true clinical thrombophilia; reasons for this interpretation are discussed. Contrary to other published non-population-based studies, we have not shown an association between thrombophilia and hemiplegic CP. More studies, including maternal studies, are required to explore this complex subject further. PMID- 11730145 TI - Video-EEG in the diagnosis of paroxysmal events in children with mental retardation and in children with normal intelligence. AB - Video-EEG monitoring was performed to evaluate 193 children (91 females, 102 males; mean age 9.6 years, SD 5.7) who presented with paroxysmal events of uncertain etiology. Diagnosis of the type of event, i.e. epileptic or non epileptic, was successfully established in 130 of 193 patients (67.3%). Seventy children (36%) had mental retardation* (MR). Children with MR were more likely (p<0.05) than children without MR to have events during the studies. Children with and without MR had strikingly similar frequencies of epileptic and non epileptic events. In participants who had events recorded and characterized, epileptic seizures were identified in 67 children (51.5%), non-epileptic events in 54 children (41.5%), and both epileptic and non-epileptic events in nine children (7%). Improved diagnosis prompted appropriate management. This should encourage more frequent use of video-EEG in children, especially in those with MR, to differentiate epilepsy from behavioral disturbances so that specific treatment can be provided. PMID- 11730146 TI - Melatonin is useful for recording sleep EEGs: a prospective audit of outcome. AB - The study aimed to determine if melatonin could reliably induce sleep in children undergoing sleep EEG without affecting the usefulness of the EEG itself. One hundred and sixty three children (112 males, 51 females; mean age 8 years, range 1 to 16 years) referred for sleep EEG were studied. The children were given 2 to 10 mg of melatonin, depending on age, just before EEG recording. Measurements included sleep-onset latency, adverse effects, and acceptability of the EEG. Usefulness and acceptability of melatonin-induced sleep EEG were compared with the standard technique of sleep EEG following sleep deprivation in 30 children (matched for sex and age). Sleep was obtained in 79% of the 163 children who received melatonin after an average of 33 minutes. Yield of epileptiform abnormalities demonstrated in the melatonin sleep EEG was similar to that reported in the literature for sleep-deprived EEGs. There was no significant adverse effect. When compared, a melatonin-induced sleep EEG was as useful as a sleep-deprived EEG. However, the children's behaviour on the day of the melatonin induced sleep EEG recording was more acceptable to parents. PMID- 11730147 TI - Sign language in childhood epileptic aphasia (Landau-Kleffner syndrome). AB - Acquired epileptic aphasia (AEA, or Landau-Kleffner syndrome) is a unique condition in which children can lose oral language (OL) comprehension and expression for a prolonged period. These children can benefit from visual forms of language, mainly sign language (SL), but the quality of SL has never been analyzed. The case is reported here of a boy with AEA who lost speech comprehension and expression from 3 years 6 months to 7 years and was educated in SL from the age of 6 years. His SL was evaluated at the age of 13 years and 6 months and compared with a control child with congenital sensorineural deafness. It was found that: (1) our patient achieved the same proficiency in SL as the control child with deafness; (2) SL learning did not compete with, but perhaps even hastened, the recovery of OL. Intact ability to learn a new linguistic code such as SL suggests that higher-order language areas were preserved and received input from a separate visual route, as shown by neuropsychological and functional imaging research in deaf and hearing signers. PMID- 11730148 TI - Self-injurious behaviour in young children with Lesch-Nyhan syndrome. AB - The early development of self-injurious behaviour in three young boys (aged 17, 25, and 30 months at start of study) with Lesch-Nyhan syndrome was examined by means of parental interviews and by direct observations completed at 3 to 4 monthly intervals over an 18-month period. Results suggest that the self-injury began in a different way from that of other young children with autism and/or developmental disabilities in that, from the start, self-injurious responses were sudden and violent, rather than emerging gradually over time. Drastic measures, such as removal of the teeth or provision of tooth guards, were often taken to prevent further tissue damage. Direct observations showed that the boys' self injury occurred at lower rates, but their carers were highly concerned about the behaviour. Sequential analysis of the observational data indicated that on some occasions the children were more likely to self-injure during periods of low social interaction, suggesting that their self-injury may have been influenced by environmental factors. The theoretical and practical implications of these findings are discussed. PMID- 11730149 TI - Autistic disorders in Down syndrome: background factors and clinical correlates. AB - A study of a clinic-based sample of 25 individuals (12 females, 13 males; age at diagnosis 14.4 years, SD 7.4 years; age range 4 to 33 years) with Down syndrome (DS) and autism spectrum disorders, demonstrates that autism is by no means rare in DS. Results showed that there was a considerable delay in the diagnosis of autism as compared with children with autism who did not have DS. In 11 participants medical factors were identified that were likely to be of importance in contributing to the development of autism, and in four further participants there were factors of possible significance. Such factors include a history of autism or autism-related disorders in first- or second-degree relatives (n=5), infantile spasms (n=5), early hypothyroidism (n=3), evidence of brain injury after complicated heart surgery (n=2), or a combination of these factors. It is important that autism is recognised, identified, and fully assessed in individuals with DS in order for them to receive appropriate education and support. PMID- 11730150 TI - Is increased D2 receptor availability associated with response to stimulant medication in ADHD. AB - The purpose of this study was to estimate striatal dopamine (D2) receptor availability in non-drug treated children with attention-deficit-hyperactivity disorder (ADHD) before and after methylphenidate therapy, and to examine correlations between severity of symptoms and response rates to stimulant medication with levels of striatal D2 receptor binding. Nine children (six males, three females; mean age 9.8 years, SD 2.3 years) with ADHD participated. All underwent iodobenzamide (123I IBZM) brain SPECT within 2 hours following intravenous injection of 123I IBZM before and 3 months after methylphenidate therapy. A semiquantitative approach was used to generate indices of specific D2 receptor binding in the basal ganglia. Specific binding ratios at baseline were higher than the previously reported specific binding values obtained in studies using young healthy adults. D2 availability reduced significantly (paired t test,p<0.05) as a function of methylphenidate therapy in patients with ADHD in all four regions of the striatum. When the relation between therapy response and D2 availability was investigated, we observed that the higher the baseline D2 levels were, the higher the response rate was (detected as the percentage reduction of hyperactivity scores and Conners Teacher Rating Scale scores), while no such trend was observed between the initial D2 binding levels and the response in attention-deficit scores. Results indicate that in non-drug treated children with ADHD, higher D2 receptor availability is observed at baseline which is down regulated back to reported near-normal values after methylphenidate therapy. The effect of methylphenidate on D2 receptor levels in patients with ADHD is similar to that observed in healthy adults; a down-regulation phenomenon within 0 to 30%. In addition, initially higher values of D2 availability seem to indicate better response to methylphenidate therapy in ADHD. PMID- 11730151 TI - Reliability of the Melbourne assessment of unilateral upper limb function. AB - This study examines the reliability of the Melbourne Assessment of Unilateral Upper Limb Function: a quantitative test of quality of movement in children with neurological impairment. The assessment was administered to 20 children aged from 5 to 16 years (mean age 9 years 10 months, SD 2 years 10 months) who had various types and degrees of cerebral palsy (CP). The performances of the 20 children during assessment were videotaped for subsequent scoring by 15 occupational therapists. Scores were analyzed for internal consistency of test items, inter- and intrarater reliability of scorings of the same videotapes, and test-retest reliability using repeat videotaping. Results revealed very high internal consistency of test items (alpha=0.96), moderate to high agreement both within and between raters for all test items (intraclass correlations of at least 0.7) apart from item 16 (hand to mouth and down), and high interrater reliability (0.95) and intrarater reliability (0.97) for total test scores. Test-retest results revealed moderate to high intrarater reliability for item totals (mean of 0.83 and 0.79) for each rater and high reliability for test totals (0.98 and 0.97). These findings indicate that the Melbourne Assessment of Unilateral Upper Limb Function is a reliable tool for measuring the quality of unilateral upper limb movement in children with CP. PMID- 11730152 TI - Powered wheelchairs and independence in young children with tetraplegia. AB - The aim of this study was to assess the effect of early provision (< or = 8 years) of a powered wheelchair (PWC) in children with tetraplegia. Twenty-nine children (15 males, 14 females; mean age 6 years 3 months, age range 3 to 8 years) with spastic or dystonic tetraplegia were studied. All participants had severe motor impairment. Treatment outcomes were investigated in several dimensions of disablement: Impairment, Functional Limitation/Activity, Disability/Participation. It was found that the level of independence improved significantly after PWC provision, while motor impairment, IQ, and quality of life did not. The majority of children (21 of 27) reached a level of driving competence which allowed them to move around with or without minimal (i.e. verbal) adult support. Achievement of this competence was not statistically related to IQ or motor impairment but correlated to the time spent in the PWC. The majority of parents (21 of 25) were not in favour of the PWC when the study started but after PWC provision, 23 of 25 parents expressed positive feelings about it. Reactions of the majority of children (23 of 25) were positive from the beginning of the study and did not change over time. The authors concluded that PWCs can aid independence and socialization and the majority of children can achieve a good-enough driving competence, even those with severe learning disability or motor deficit. PWCs should not be viewed as a last resort but as a means of providing efficient self-locomotion in children with a severe motor deficit. PMID- 11730153 TI - Effects of neurodevelopmental treatment (NDT) for cerebral palsy: an AACPDM evidence report. PMID- 11730154 TI - 'Adverse events following repeated injections with botulinum toxin A in children with spasticity'. PMID- 11730155 TI - 'Life expectancy among people with cerebral palsy in Western Australia'. PMID- 11730156 TI - Bacteriophages as therapeutic agents. AB - The emergence of antibiotic resistance among pathogenic bacteria is one of the most critical problems of modern medicine, and novel, effective approaches for treating infections caused by multidrug-resistant bacteria are urgently required. In this context, one intriguing approach is to use bacteriophages (viruses that kill bacteria) to eliminate specific bacterial pathogens. Bacteriophage therapy was widely used around the world in the 1930s and 1940s, and it is still used in Eastern Europe and the former Soviet Union. However, phage therapy was all but abandoned in the West after antibiotics became widely available. Promising results from recent animal studies using phages to treat bacterial infections, together with the urgent need for novel and effective antimicrobials, should prompt additional rigorous studies to determine the value of this therapeutic approach. PMID- 11730157 TI - Intracerebral regulation of immune responses. AB - Major progress has been made over the last years in our understanding of the mechanisms underlying immune privilege and immune surveillance of the central nervous system (CNS). Once considered a passive process relying only on physical barriers, immune privilege is now viewed as a more complex phenomenon, which involves active regulation of immune reactivity by the CNS microenvironment. Evidence has also emerged that the immune system continuously and effectively patrols the CNS and that dysregulated immune responses against CNS-associated (exogenous or self) antigens are involved in the pathogenesis of various neurological diseases. In this article we shall briefly review current knowledge of how the immune response is regulated locally in the CNS and which cell types and molecular mechanisms are involved in shaping intracerebral immune responses. PMID- 11730158 TI - Thalidomide treatment for refractory Crohn's disease: a review of the history, pharmacological mechanisms and clinical literature. AB - Several recent case reports and clinical trials have demonstrated that thalidomide is emerging as an efficacious alternative in the treatment of selected patients with refractory Crohn's disease. The effects of thalidomide are at least partly mediated by down-regulation of tumour necrosis factor (TNF) alpha, a potent proinflammatory cytokine. However, thalidomide is also known to inhibit angiogenesis, and it has several other well-described immunomodulatory properties. Clinical studies have confirmed that previously refractory Crohn's disease patients respond to thalidomide, and many enter clinical remission. Efficacy usually occurs within 4 weeks. Thalidomide also has steroid-sparing properties, and it is particularly useful in treating oral and fistulous complications of Crohn's disease. Although it is usually tolerable, careful monitoring is recommended to prevent toxicities, such as birth defects and peripheral neuropathy. This review provides a detailed summary of the literature to date on the use of thalidomide treatment for Crohn's disease. Special attention is directed towards its history, mechanisms, and proposed role. The recent development of thalidomide analogues is also discussed briefly. PMID- 11730159 TI - Genetic kidney diseases disclose the pathogenesis of proteinuria. AB - The sieving of plasma components occurs in the kidney through the glomerular capillary wall. This filter is composed of three layers: endothelium, glomerular basement membrane (GBM), and podocyte foot processes connected by slit diaphragms. Defects in this barrier lead to proteinuria and nephrotic syndrome. Previously, defective GBM was regarded to be responsible for proteinuria. However, recent work on genetic diseases has indicated that podocytes and the slit diaphragm are crucial in restricting protein leakage. Congenital nephrotic syndrome of the Finnish type (NPHS1) is caused by mutations in a novel NPHS1 gene, which encodes for a cell adhesion protein, nephrin. This protein is synthesized by podocytes, and seems to be a major component of the slit diaphragm. In severe NPHS1, lack of nephrin leads to missing slit diaphragm. The role of nephrin in acquired kidney diseases remains unknown. In addition to nephrin, other podocyte proteins (podocin, alpha-actinin-4, CD2AP, FAT) have recently been identified and associated with the development of proteinuria. It seems that the slit diaphragm and its interplay with the podocyte cytoskeleton is critical for the normal sieving process, and defects in one of these components easily lead to proteinuria. PMID- 11730160 TI - Health consequences of visceral obesity. AB - Visceral obesity is associated with metabolic abnormalities that increase the risk of type 2 diabetes and coronary heart disease. Obese patients with a substantial accumulation of visceral adipose tissue are characterized by higher insulinaemic and glycaemic responses during an oral glucose challenge as well as by a deteriorated plasma lipoprotein-lipid profile compared with individuals with normal body weights or obese subjects with low levels of visceral adipose tissue. Results of the Quebec Cardiovascular Study have shown that the cluster of metabolic disturbances observed among subjects with visceral obesity (hyperinsulinaemia, hyperapolipoprotein B and small, dense low-density lipoprotein (LDL) particles) is associated with a 20-fold increase in the risk of coronary heart disease in a sample of middle-aged men followed over 5 years. Therefore, we have developed a simple screening approach in order to help physicians and health professionals identify at low cost individuals who would be characterized by this cluster of atherogenic abnormalities. We found that the simultaneous presence of an elevated waist girth combined with moderate hypertriglyceridaemia ('hypertriglyceridaemic waist') could adequately identify a large proportion (approximately 80%) of carriers of the above triad of atherogenic metabolic abnormalities (hyperinsulinaemia, hyperapolipoprotein B and small, dense LDL particles). Finally, there is evidence suggesting that the risk of an acute coronary syndrome in these viscerally obese patients may not always be related to the extent of coronary artery stenosis, providing further support to the notion that additional markers of thrombosis/inflammation should be considered. Thus, the stabilization of the atherosclerotic plaque, rather than its regression may even become a more legitimate and feasible therapeutic objective for the management of the coronary heart disease risk in the viscerally obese patient. Although these notions are based on a plausible metabolic rationale, randomized trials with proper end-points will be needed to determine the clinical benefits associated with the management of visceral obesity and related metabolic complications. PMID- 11730161 TI - Genetic variance and lipolysis regulation: implications for obesity. AB - Catecholamines are the major lipolytic hormones in human fat cells, and lipolytic catecholamine resistance is described in obesity. Studies on twins and in rare genetic disorders suggest a strong heredity component of catecholamine-induced lipolysis. Polymorphisms in catecholamine receptor signalling pathways have been described, several of which associate with obesity. Many polymorphisms in adrenoceptor genes are functional in transfected cell lines. The importance of polymorphisms in catecholamine signalling pathways for lipolysis regulation is discussed. A Trp64Arg polymorphism in the beta3-receptor, which associates with obesity, is accompanied by changes in lipolytic sensitivity of the receptor in human fat cells. Similarly, a Gln16Glu and an Arg164Ile variation in the beta2 adrenoceptor cause marked variations in the lipolytic sensitivity of this receptor in human adipocytes. Furthermore, beta2-adrenoceptor gene polymorphisms associate with obesity. A dinucleotide (CA) intron repeat in hormone-sensitive lipase gene is linked to obesity and markedly decreases the ability of catecholamines to activate the lipase and thereby lipolysis in human fat cells. However, an Arg389Gly polymorphism in the beta1-adrenoceptor, which alters receptor function in transfected cell lines, has no effect on lipolysis in human fat cells and is not associated with obesity. Thus, polymorphism in human genes that are involved in catecholamine signal transduction have effects on fat cell lipolysis and also relate to obesity. The lipolysis effects of these polymorphisms cannot always be predicted from gene transfer experiments on artificial cell lines. It is possible that genetic variance in catecholamine signalling pathways, through alterations in adipocyte lipolysis, may promote obesity. PMID- 11730162 TI - Gene expression in visceral and subcutaneous adipose tissues. AB - A large body of evidence demonstrates depot-specific differences in the expression of genes coding important functional proteins in adipocytes. This may contribute to the well-known specific functional properties of the adipocytes from intra-abdominal and subcutaneous regions. This review will focus on the main findings regarding the regional differences in adipocyte gene expression in humans. These genes encode proteins belonging to three different functional groups: the metabolic enzyme and related signalling proteins, the adipogenic factors, and, finally, the products of adipocytes. PMID- 11730163 TI - Regulation of adipocyte differentiation. AB - Once multipotent mesenchymal cells become committed to the adipoblast lineage, adipogenesis, the process of preadipocytes differentiation into adipocytes is initiated. This process starts with a phase of exponential growth of adipoblasts. Following confluence of these adipoblasts, the cells enter into a cell cycle arrest, they re-enter the cell cycle and pass through a limited number of cell divisions, and finally differentiate into fully mature adipocytes. Adipogenesis is controlled by a complex cross-talk between positive and negative regulators, such as hormonal and nutritional stimuli, that change the activity of a selected set of transcription factors. Regulation of adipogenesis is crucial to keep the body energy balance because a limited amount of adipose tissue, lipodystrophy, or an excess of adipose tissue, such as occurs in obesity, lead to profound metabolic dysfunctions and disease. PMID- 11730164 TI - 'Dangerous dogs' on the agenda at the FECAVA congress in Berlin. PMID- 11730165 TI - Rapid test for the preclinical postmortem diagnosis of BSE in central nervous system tissue. AB - The efficacy of a rapid test for detecting PrP(Sc) in central nervous system tissue was evaluated for the postmortem diagnosis of BSE at different times during the course of the disease. One hundred and six samples of brain, at the level of the medulla oblongata, and spinal cord, derived from the experimental study of the pathogenesis of BSE carried out in Great Britain between 1991 and 1995, were examined. PrP(Sc) was detected in the samples from most of the exposed animals killed 32 months or more after they had been exposed to the agent, and before the onset of clinical signs which were first recorded at 35 months. Comparisons with the results of histology, fibril detection, PrP immunohistochemistry and mouse bioassay indicated that the rapid test is at least as sensitive as these conventional confirmatory diagnostic methods and its result can be obtained more quickly. PMID- 11730166 TI - Safety and efficacy of an oil-adjuvant vaccine against haemorrhagic septicaemia in buffalo calves: cross-protection between the serotypes B:2,5 and E:2,5. AB - The safety, efficacy and duration of immunity of an improved oil-adjuvant vaccine against haemorrhagic septicaemia, containing inactivated cells of Pasteurella multocida serotype B:2,5, were tested in young buffalo calves in Pakistan. For safety testing, five buffalo calves were vaccinated intramuscularly with twice the normal dose, and six weeks later with a normal dose. Except for a transient rise in rectal temperature at six hours after the vaccinations, no systemic reactions were observed. The buffaloes remained in good condition and had a normal appetite. No local reactions were observed at the injection site. For efficacy testing two trials were carried out. In the first, buffalo calves were vaccinated intramuscularly either with two doses two-and-a-half months apart, or with a single dose, or left unvaccinated. They were challenged subcutaneously with virulent P multocida after eight, 13 or 15 months. After challenge at eight months the four buffaloes given two doses and the buffalo given one dose were protected, whereas the control animal developed the typical signs of the disease. After the challenges at 13 and 15 months, the vaccinated animals were still protected whereas the control animals died. In the second trial, buffalo calves were vaccinated intramuscularly either with two doses two months apart, or with a single dose at two months or left unvaccinated. The buffaloes were challenged after eight or 14 months. After challenge at eight months the four control animals died, whereas three of the four buffaloes given a single dose were protected. After challenge at 14 months, the three control animals died, whereas four of the five buffaloes given two doses and both the buffaloes given a single dose were protected. To test for cross-protection against the heterologous serotypes E:2,5 and B:3,4, groups of mice were vaccinated once or left unvaccinated. Four weeks later, the vaccinated and control groups were challenged with a dilution series of the different challenge cultures. The vaccine appeared to induce protection against challenge with different strains of serotypes B:2,5 and E:2,5 but not against strains of serotype B:3,4. PMID- 11730167 TI - Intraocular pressure of juvenile loggerhead sea turtles (Caretta caretta) held in different positions. AB - The intraocular pressure of 12 apparently healthy juvenile loggerhead sea turtles (Caretta caretta) was determined by applanation tonometry while the turtles were held in dorsoventral, ventrodorsal, and head-down suspended positions. The median intraocular pressures were 5 mmHg (range 4 to 9 mmHg) in the dorsoventral position, 7 mmHg (range 5 to 12 mmHg) in the ventrodorsal position, and 23 mmHg (range 17 to 33 mmHg) in the suspended position. PMID- 11730168 TI - Modulation of prolactin hormone and intersequence pause days in domestic chickens. PMID- 11730169 TI - Lateral meniscal tears and stifle osteochondrosis in three dogs. PMID- 11730170 TI - Aggressive angiomyxoma in a cow. PMID- 11730171 TI - Filariasis in wild-caught chameleons. PMID- 11730172 TI - Vigilance against bioterrorism. PMID- 11730173 TI - Trauma-induced severe lameness in calves. PMID- 11730174 TI - Modelling the FMD outbreaks. PMID- 11730175 TI - Monitoring mastitis and maintaining biosecurity. PMID- 11730176 TI - Adrenal suppression with inhaled corticosteroids. PMID- 11730177 TI - The immunologic workup of the child suspected of immunodeficiency. AB - OBJECTIVE: This review is intended to provide an outline for the evaluation of patients suspected of having immunodeficiency, a problem that is frequently encountered in clinical practice. DATA SOURCES: Information was obtained through a MEDLINE literature search as well as from standard textbooks in immunology. Also included is information that reflects the authors' clinical experience in the field. RESULTS: In general clinical practice, many physicians feel inadequate to evaluate patients with suspected immune deficiencies. They also think that the process of evaluation is time-consuming, which results in misdiagnosis of a substantial percentage of such disorders. Hence, the prevalence of immunodeficiency disorders is much higher than generally thought. At present, there are >80 unique primary immunodeficiency conditions and >50 syndromes that are associated with various immunologic defects. The prevalence of secondary immunodeficiency has also been increasing because of the tragic epidemic of HIV infection, more usage of immunosuppressive medications and bone marrow stem cell transplantation, and the severe degree of malnutrition in underdeveloped countries. It is necessary for clinicians, particularly the specialists in allergy and immunology, to be able to evaluate the status of the immune system. CONCLUSIONS: Very valuable information can be gathered from the medical history and physical examination that may exclude or increase the suspicion of immunologic defect. Laboratory tests can then be appropriately selected to define the specific defect. Once the diagnosis has been settled, proper medical management can be instituted with subsequent improvement in morbidity and mortality of such disorders. PMID- 11730178 TI - Selective CD4+ T cell lymphocytopenia and recalcitrant warts in an 8-year-old child. PMID- 11730179 TI - Suppression of hypothalamic-pituitary-adrenal axis activity with inhaled flunisolide and fluticasone propionate in adult asthma patients. AB - BACKGROUND: Suppression of the hypothalamic-pituitary-adrenal (HPA) axis, a potential systemic effect of inhaled corticosteroid therapy, can be quantified by monitoring serum, urinary, and salivary cortisol levels. OBJECTIVES: 1) Compare the effects on HPA axis of the inhaled corticosteroids flunisolide and fluticasone propionate versus placebo and oral prednisone. 2) Estimate dose potency ratio for HPA-axis suppression. METHODS: Multicenter, randomized, placebo controlled, open-label, 21-day trial. Active regimens were flunisolide 500 and 1,000 microg, twice daily; fluticasone propionate 110, 220, 330, and 440 microg, twice daily; and prednisone, 7.5 mg daily. Enrolled patients were nonsmokers, 18 to 50 years of age, with persistent mild-to-moderate asthma and had not used oral, nasal, or inhaled corticosteroids for 6 months before study. Main outcome measures were area under serum cortisol concentration curve for 22 hours (AUC(0 22h)); 24-hour urinary cortisol level; and 8 AM salivary cortisol level. RESULTS: One hundred fifty-three patients were randomly assigned to active treatment or placebo; 125 patients completed the study and were at least 80% compliant with their regimens. Both fluticasone propionate and flunisolide caused dose-dependent suppression of HPA axis, which was statistically greater for fluticasone propionate (P = 0.0003). Dose-potency ratio showed 4.4 times more serum-cortisol suppression/microgram increase in dose with fluticasone propionate than with flunisolide. Diurnal pattern of serum cortisol suppression was persistent with fluticasone propionate and "remitting" with flunisolide. Salivary and urinary cortisol data were qualitatively similar to serum cortisol results. CONCLUSIONS: Fluticasone caused significantly more suppression of HPA axis than flunisolide. Flunisolide may provide a safe option for patients with asthma requiring long term inhaled corticosteroid therapy. PMID- 11730180 TI - Skin prick test in duplicate: is it necessary? AB - BACKGROUND: Duplicate skin prick testing has previously been recommended because of reports that accidental negative tests are common. However, duplicate tests also mean an extra allergen load, which may increase the risk of inducing a generalized reaction at the test situation, at least in the youngest infants. OBJECTIVE: To investigate whether the occurrence of both a positive and negative test result is a common feature when performing duplicate skin prick tests and can therefore justify the duplicate method. METHODS: A retrospective analysis of all skin prick tests performed in duplicate at the pediatric clinic at University Hospital in Linkoping, Sweden, in 1997. RESULTS: Of 1,087 skin prick tests, 14 resulted in one positive and one negative test, or 1.3%. The corresponding figure in the youngest age group, (ie, <2 years of age) was 3 of 340 (0.9%). CONCLUSIONS: Considering the risk of inducing a summation of the reactions, and thereby a generalized allergic reaction, when applying an extra allergen load on the limited surface of the small arm, we conclude that the results of this study justify using single prick test, at least in the youngest age group and probably when testing children of all ages. PMID- 11730181 TI - A rapid, reproducible method for coating Rotorod Sampler collector rods with silicone grease. AB - BACKGROUND: The Rotorod Sampler (Sampling Technologies, St. Louis Park, MN) is a rotating-arm impactor that recovers airborne particles on two rapidly moving plastic collector rods. For decades, the standard method for applying silicone grease to collector rods has been with one's finger. Although this method can yield excellent results when performed by practiced investigators, a relatively high skill level is required, and significant intrapreparer variability has been reported in the medical and technical literature. OBJECTIVE: The purpose of this investigation was to develop and evaluate a new method for coating Rotorod collector rods with silicone grease. METHODS: Collector rods were coated with silicone grease by dipping them into a solution consisting of silicone grease and hexane. Pollen recovery by these dipped collector rods was compared with pollen counts obtained with hand-greased collector rods. RESULTS: Twenty-three paired samples were obtained during five sampling periods. Pollen recovery by the hand greased and dipped collector rods was similar (P = 0.410). Dipped collector rods generally offered a lower standard deviation than hand-greased collector rods, however, differences were not statistically significant (F = 1.782, P = 0.087). Dipped collector rods were also superior to hand-greased collector rods in several qualitative categories such as grease uniformity, time required for microscopic analysis, and visual quality. CONCLUSIONS: Dipped collector rods offered a time-efficient means to obtain atmospheric samples with excellent visual quality. The resulting pollen counts were similar to data obtained via the standard, manual method. Allergists are encouraged to consider using this new method in their office practices and for drug studies. PMID- 11730182 TI - A comparative, volumetric survey of airborne pollen in Philadelphia, Pennsylvania (1991-1997) and Cherry Hill, New Jersey (1995-1997). AB - BACKGROUND: Aeroallergen sampling provides information regarding the onset, duration, and severity of the pollen season that clinicians use to guide allergen selection for skin testing and treatment. OBJECTIVE: This atmospheric survey reports: 1) airborne pollen contributors in Philadelphia, Pennsylvania (1991 to 1997) and suburban Cherry Hill, New Jersey (1995 to 1997); 2) pollen onset, duration, and peak level; and 3) the relationship between airborne pollen and selected meteorologic variables. METHODS: At both locations, separated by 11 km. sampling was performed with a volumetric Rotorod Sampler (Sampling Technologies, Minnetonka, MN). RESULTS: In Philadelphia and Cherry Hill, respectively, 3-year average measurements included 75.0 and 74.2% tree pollen, 10.2 and 8.3% grass pollen, and 14.8 and 17.5% weed pollen. Prominent airborne pollen taxa were Acer, Quercus, Betula, Pinus, Cupressaceae, Poaceae, Ambrosia, and Rumex. The tree, grass, and weed pollen seasons extended from mid-March to mid-June, late April to mid-June, and mid-August to late September, respectively. A secondary Poaceae pollen peak occurred in September. There was a statistically significant correlation between simultaneous weekly average pollen levels in Philadelphia and in Cherry Hill (Acer, r(p) = 0.987, Quercus, r(p) = 0.645, Betula, r(p) = 0.896, Pinus, r(p) = 0.732, Cupressaceae, r(p) = 0.695, Poaceae, r(p) = 0.950, Ambrosia, r(p) = 0.903, and Rumex, r(p) = 0.572, P <0.001). Daily Poaceae pollen levels were positively influenced by same-day high temperature (r(s) = 0.333 in Philadelphia and r(s) = 0.426 in Cherry Hill, P < 0.05). Daily Ambrosia pollen levels were inversely influenced by same-day total precipitation (r(s) = -0.174 in Philadelphia and r(s) = -0.257 in Cherry Hill, P < 0.05). CONCLUSIONS: This is the first volumetric survey performed in either Philadelphia or Cherry Hill. Copious amounts of airborne pollen were seen from late April to early May and in early September. Pollen onset, duration, and year-to-year variability were similar at both sites. An awareness of local aeroallergen patterns is critical in the effective testing and treatment of atopic individuals. PMID- 11730183 TI - Effective control of asthma with hydrofluoroalkane flunisolide delivered as an extrafine aerosol in asthma patients. AB - BACKGROUND: Inhaled corticosteroids are established as maintenance therapy for persistent asthma. A new aerosol formulation of flunisolide delivers a small particle size by using a hydrofluoroalkane (HFA) propellant with a built-in spacer. OBJECTIVE: To compare efficacy and safety of two different flunisolide formulations, HFA and chlorofluorocarbon (CFC), with placebo treatment over a range of doses. METHODS: The multicenter, randomized, double-blind, placebo controlled trial consisted of a 2-week, active run-in phase with CFC flunisolide 500 microg, twice daily, followed by 12 weeks of double-blind treatment with placebo, HFA flunisolide (85, 170, or 340 microg, twice daily), or CFC flunisolide (250, 500, or 1,000 microg, twice daily). Patients (N = 669) were nonsmokers, at least 12 years of age, with mild to moderate asthma who were being treated with inhaled corticosteroids. Outcome measures were change from baseline in forced expiratory volume in 1 second (FEV1), peak expiratory flow rate, as needed albuterol use, nocturnal awakenings, and asthma symptoms. RESULTS: After 12 weeks of treatment, patients receiving 170 microg, twice daily, and 340 microg, twice daily, of HFA flunisolide showed a significant (P < 0.01) improvement in percentage increase in FEV1 (12.22% at 170 microg, twice daily, and 14.69% at 340 microg, twice daily) compared with the placebo group (5.35%). At one-third the dose of CFC flunisolide, HFA flunisolide provided similar improvement in pulmonary function versus placebo. Both formulations demonstrated comparable linear dose dependency for the change from baseline in FEV1 without any evidence of cortisol suppression. Outcome values for all seven secondary efficacy measures were numerically superior in patients receiving HFA flunisolide compared with the CFC formulation. Both formulations seemed to be safe and well tolerated. CONCLUSIONS: HFA flunisolide provides comparable efficacy and safety at one-third the dose of CFC flunisolide. PMID- 11730184 TI - Interleukin 6 and C-reactive protein levels in patients with acute allergic reactions: an emergency department-based study. AB - BACKGROUND: Elevations of interleukin 6 (IL-6) have been described in drug induced anaphylaxis. Although IL-6 is well known to stimulate an acute phase response, profiling acute phase protein levels, such as C-reactive protein (CRP), has, to our knowledge, never been performed in patients with acute allergic reactions. OBJECTIVE: To examine the pattern of IL-6 and CRP levels in patients with acute allergic reactions and to relate these to relevant clinical and laboratory parameters. METHODS: Plasma CRP and serum IL-6 levels were determined in 85 adult emergency department patients. These patients had been previously studied with questionnaires, physical examinations, and histamine/tryptase levels. Clinical and historical features were related to CRP and IL-6 levels. CRP and IL-6 levels were also examined for relationships with histamine and tryptase levels. RESULTS: CRP and IL-6 levels were significantly correlated with one another in the study patients (Spearman p = 0.36, P = 0.0008). Similar to histamine levels, IL-6 levels were significantly correlated with the extent of erythema manifested by the study patients. The extent of erythema was independently predicted by both IL-6 and histamine levels. Histamine levels were negatively correlated with CRP levels (Spearman p = -0.32, P = 0.003). Unlike histamine levels, IL-6 and CRP did not show significant relationships with the extent or presence of urticaria/angioedema or the presence of wheezing. IL-6 levels were correlated with the duration of symptoms before serologic sampling. An inverse correlation was observed between IL-6 levels and mean arterial blood pressure. Multivariate modeling showed significant independent effects from mean arterial pressure, duration of symptoms, erythema extent, and age in predicting IL-6 levels. Tryptase levels were higher in patients whose IL-6 levels were >20 pg/mL. CONCLUSIONS: CRP and IL-6 levels are not simple surrogate markers for histamine or tryptase release by mast cells or basophils in acute allergic reactions. Increasing IL-6 levels relate to greater erythema extent, lower mean arterial blood pressure, and a longer duration of symptoms. It would be interesting to speculate that CRP and IL-6 increases characterize a late-phase response in immediate hypersensitivity reactions. In this perspective, the inverse relationship between CRP and histamine levels could be explained. As histamine levels are waning, CRP levels are increasing. Timed studies for histamine and CRP/IL-6 levels in allergic reactions are necessary to confirm this hypothesis. PMID- 11730185 TI - The misconception of using floating patterns as an accurate means of measuring the contents of metered-dose inhaler devices. AB - BACKGROUND: Patients and physicians have searched for a reproducible method of determining the amount of medication that a metered-dose inhaler (MDI) contains as well as a reliable method of determining when their MDI is empty. Previously, patients have been instructed to float their canister in water, and depending upon the position attained, have been able to estimate the amount of medication within the canister. OBJECTIVE: To investigate whether the floating patterns of MDIs are a reliable method of determining the contents contained within an inhaler canister, including that of the newer devices containing the non chlorofluorocarbon (CFC) propellant. METHODS: Fifteen albuterol sulfate MDIs (Proventil HFA; Schering, Kenilworth, NJ), 15 triamcinolone acetonide MDIs (Azmacort; Rhjne-Poulenc Rorer, Collegeville, PA), and 15 fluticasone propionate MDIs (Flovent; GlaxoSmith Kline, Research Triangle Park, NC) were obtained from their respective companies. Each device was floated in a clear container full of water before any actuations. The devices were then actuated into the air at 2 minute intervals and each subsequently floated following 25%, 50%, 75%, and 100% of the prescribed number of actuations and its position within the container observed. The canisters were then actuated until no visual spray was produced from the nozzle and again their floating positions within the container observed. RESULTS: Each of the three MDIs tested had unique floating patterns both before any actuation as well as throughout the various actuations. CONCLUSIONS: This study demonstrates that the floating method is not an accurate means by which patients can identify the amount of medication contained within an inhaler device. This includes both conventional MDIs containing CFC propellant, as well as the newer non-CFC MDIs. PMID- 11730186 TI - Effect of salmeterol and salmeterol plus beclomethasone on saliva flow and IgA in patients with moderate-persistent chronic asthma. AB - BACKGROUND: The use of short-acting beta2-agonists is associated with oral mucosa injuries that are probably provoked by decreased saliva flow and decreased concentrations of immunoglobulin (Ig)A in saliva. OBJECTIVES: To explore the effect of salmeterol, alone or combined with beclomethasone, on the health of oral mucosa, as well as its effect on saliva flow and IgA concentration in saliva. METHODS: Patients ranging in age from 6 to 15 years with moderate persistent chronic asthma were enrolled. Patients received two 6-week treatments, one with salmeterol plus beclomethasone and the other with only salmeterol, with a 1-week washout period between treatments. Patients had oral cavity examinations and assessments of saliva flow, IgA in saliva, and total protein in saliva before the beginning and at the end of each treatment RESULTS: The results of the baseline oral examinations were normal in all patients. The postsalmetrol (PS) examinations detected 13 patients with gingivitis and the postbeclomethasone salmeterol (PBS) examinations disclosed 10 patients with gingivitis and 1 with lower-lip ulceration. Baseline saliva flow was 16.25 +/- 7.04 mm/minute (confidence interval [CI] 95% 13.67; 18.89), PS was 13.53 +/- 5.93 mm/minute (CI 95% 11.33; 15.73), and PBS was 16.57 +/- 5.54 mm/minute (CI 95% 14.51; 18.62). No statistical differences between the different assessments were found. Mean saliva IgA at baseline was 4.99 +/- 1.96 mg/dL (CI 95% 4.26; 5.71), PS IgA was 6.53 +/- 3.02 mg/dL (CI 95% 5.41; 7.65), and PBS IgA was 4.82 +/- 1.98 mg/dL (CI 95% 4.08; 5.56). PS IgA was significantly higher than the other two determinations (P < 0.05 by Bonferroni and Tukey tests). Baseline saliva IgA-to-protein ratio was 0.72 +/- 0.24 (95% CI 0.64; 0.80), PS IgA:protein ratio was 1.02 +/- 0.38 (95% CI 0.88; 1.16), and PBS IgA:protein ratio was 0.72 +/- 0.25 (95% CI 0.62; 0.82). PS IgA:protein ratio was significantly higher than the other two determinations (P < 0.05 by Bonferroni and Tukey tests). CONCLUSIONS: In the present study it was demonstrated that salmeterol alone or in combination with beclomethasone induced injuries in the oral mucosa, but only salmeterol alone induced increases in the total and protein-adjusted IgA in saliva. PMID- 11730187 TI - Olopatadine inhibits anti-immunoglobulin E-stimulated conjunctival mast cell upregulation of ICAM-1 expression on conjunctival epithelial cells. AB - BACKGROUND: Olopatadine is a clinically effective dual-action (antihistamine/mast cell stabilizer) ophthalmic antiallergic agent. We have previously demonstrated that olopatadine inhibits tumor necrosis factor alpha (TNF-alpha) release from purified human conjunctival mast cells and that supernates from stimulated mast cells upregulate intercellular adhesion molecule 1 (ICAM-1) expression on epithelial cells via TNF-alpha. OBJECTIVE: To investigate the effect of olopatadine on the TNF-alpha-mediated mast cell upregulation of ICAM-1 expression on conjunctival epithelial cells. METHODS: Human conjunctival mast cells and epithelial cells were purified (>95%) from cadaveric tissue. Conjunctival mast cells were preincubated with three doses (30, 300, or 3,000 microM) of olopatadine or buffer alone for 30 minutes followed by 90-minute challenge with anti-immunoglobulin E (10 microg/mL). The resulting supernates were incubated with conjunctival epithelial cell monolayers for 24 hours along with the following treatments: rTNF-alpha, mast cell supernate + anti-TNF-alpha, recombinant (r)TNF-alpha + anti-TNF-alpha, the three doses of olopatadine, olopatadine supernates, olopatadine supernates + rTNF-alpha. ICAM-1 expression was measured using flow cytometry. RESULTS: Anti-IgE-stimulated human conjunctival mast cell supernates upregulated human conjunctival epithelial cell ICAM-1 expression to the same extent as rTNF-alpha. ICAM-1 upregulation could be completely blocked with anti-TNF-alpha. Preincubation of conjunctival mast cells with olopatadine significantly blocked the ability of supernates to upregulate ICAM-1 on conjunctival epithelial cells. ICAM-1 expression could be restored by adding rTNF-alpha to the olopatadine-preincubated mast cell supernates. CONCLUSIONS: Olopatadine is able to significantly decrease the anti immunoglobulin E mast cell supernate-mediated upregulation of ICAM-1 on human conjunctival epithelial cells in vitro. This seems to be mediated through an effect on a TNF-alpha-specific mechanism. PMID- 11730188 TI - Anaphylaxis to riboflavin (vitamin B2). AB - BACKGROUND: Vitamin supplements are used more commonly in normal healthy subjects than in patients with vitamin deficiency. Thiamine (vitamin B,) is the vitamin that most frequently induces allergic reactions. To the best of our knowledge, no case of anaphylaxis to riboflavin (vitamin B2) has thus far been reported in the literature. OBJECTIVE: We describe a previously healthy 15-year-old boy in whom anaphylaxis developed several times after he drank one soft drink or took a single multivitamin tablet. This study was done to determine which of the many components found in the soft drink and vitamin tablet caused the anaphylactic reaction. METHODS: In an outpatient clinic with the availability of complete resuscitative procedures, we performed single-blind prick skin tests and intradermal skin tests on the patient with various pure vitamin components of the soft drink and the multivitamin tablet. Physiologic saline and histamine were used for negative and positive controls, respectively. RESULTS: Riboflavin, a component of both the soft drink and the vitamin tablet, produced positive reactions on intradermal skin tests in the patient. Positive reactions were not present in the normal control subjects. CONCLUSIONS: Riboflavin is a previously unreported cause of anaphylaxis. Free-form riboflavin may potentially be associated with an anaphylactic reaction. It is a vitamin widely used in many patients with chronic disease and in healthy subjects. Vitamin B2 must be considered as a cause of anaphylaxis. PMID- 11730189 TI - Desensitization to urine-derived gonadotropins in a woman with secondary infertility. AB - BACKGROUND: Urine-derived gonadotropins have been used to treat infertility but may cause allergic reactions. IgE-mediated hypersensitivity reactions can be treated with desensitization, especially when new therapies such as recombinant human gonadotropins are unavailable. OBJECTIVE: This case is described to highlight a successful intervention with desensitization in a woman with a complicated history of secondary infertility. She had been treated with ovulation induction regimens, such as IVF-M (in vitro fertilization-human menopausal gonadotropin [hMG]) and IVF-C (human chorionic gonadotropin [hCG]), for intrauterine insemination. During treatment, however, she experienced reactions to IVF-M and IVF-C. Because she strongly wanted a baby and no alternative preparation was available, desensitization was attempted. METHODS: Intradermal tests with IVF-M and IVF-C using both negative and positive controls were performed on the patient and four normal control subjects. Immediate wheal-and flare reactions occurred only in the patient. ELISA and ELISA inhibition tests showed positive responses to IVF-M and IVF-C, but not to highly purified hMG, hCG, or D-mannitol, a preservative in IVF-M and IVF-C. Desensitization to IVF-M and IVF-C was done with a protocol used for parenteral desensitization to penicillin. RESULTS: During the IVF-M desensitization, the intramuscular injections were well tolerated. For the next 2 days, daily administration of IVF M was uneventful. Thirty-six hours later, desensitization to IVF-C was performed successfully. The patient had two intrauterine inseminations and became pregnant. CONCLUSIONS: Nongonadotropin proteins in urine-derived gonadotropins cause IgE mediated hypersensitivity reactions. Acute desensitization to urine-derived gonadotropins can be performed effectively, as shown in our current case. PMID- 11730190 TI - Alternaria alternata. PMID- 11730191 TI - Recovery of myocardial function with long-term ventricular assist device support. PMID- 11730192 TI - How to run an outpatient VAD program: overview. PMID- 11730193 TI - Hospital discharge for the ventricular assist device patient: historical perspective and description of a successful program. AB - Patients who are supported with an implantable ventricular assist device (VAD) as a bridge to cardiac transplantation are potential candidates for hospital discharge. Hospital discharge rates in reported clinical series vary from 27% to 60%. Many of the patients included in these series, however, were subjects of premarket approval clinical trials and as such, are bound by rigid eligibility criteria for discharge. According to a voluntary registry maintained by Thermo Cardiosystems, Inc., the postmarket discharge rate in patients supported with the HeartMate VE LVAS is approximately 25%, a number that is artificially low due to incomplete reporting. The postmarket discharge rate at the busiest Thermo Cardiosystems HeartMate VE LVAS centers is 53%. Clearly, discharge rates need to increase if the VAD is ever to be considered a viable destination therapy for end stage heart failure. The discharge program instituted at The University of Iowa incorporates patient and family training as well as community services education. Between January 1999 and April 2001, fourteen patients received VAD support as a bridge to transplantation with the HeartMate VE LVAS. Thirteen patients (93%) were discharged from the hospital. Eight of the 13 patients have been transplanted after having spent 65.8% +/- 31.4% of their period of blood pump support as an outpatient. Eight of 13 patients (62%) required a total of 20 unplanned repeat hospitalizations. A well defined, aggressively implemented discharge program can adequately prepare the VAD patient for the transition from hospital to home. PMID- 11730194 TI - Education of the ventricular assist device patient's community services. AB - Two implantable, electric left ventricular assist systems have received approval from the Food and Drug Administration (FDA) for use outside a hospital environment. As patients supported with these systems are discharged from the hospital, it is important that various community services and agencies be involved in the care of this unique patient population. Between January 1999 and May 2001, The University of Iowa Hospitals and Clinics (UIHC) implanted 14 HeartMate VE LVAS (Thoratec Corporation, Pleasanton, CA) in patients as a bridge to cardiac transplantation. Of these 14 patients, 13 were discharged home from the hospital. On the basis of our 3 year experience, a discharge program has been developed to educate the ventricular assist device (VAD) patient's community services. Ten community services most critical to a safe home discharge program are identified. Timing of service contact, mechanism of contact, community education method, education content, education materials, and outpatient community service usage are described. PMID- 11730195 TI - Potential hazards of polyglucose. PMID- 11730196 TI - Delivery of erythropoietin with a needleless injection system during hemodialysis maintains plasma levels. AB - Recent Occupational Safety and Health Administration guidelines require the implementation of needleless systems in dialysis units in the state of California. It is not known whether needleless systems deliver medication as well as traditional low dead-space needle/syringes. The purpose of this study was to compare the delivery of recombinant human erythropoietin (rHuEpo) by a needleless system (Medic Plastic Anti-Stick Needle/Connector, Medisystems, Seattle, WA) with delivery using a traditional low dead-space needle/syringe. We also studied the Medisystems High Flow venous chamber with integral injection site that is designed to prevent the formation of a separate serum layer within which rHuEpo may become trapped. There was no significant difference in the mean serum rHuEpo concentration achieved by either the needleless system, or traditional low dead space needle/syringe, whether using the venous chamber's integral injection site or the postvenous chamber in-line injection site. This study demonstrates for the first time that a needleless system of delivery can reliably achieve serum rHuEpo comparable to traditional low dead-space needle/syringes. PMID- 11730197 TI - Usefulness of the low profile "True 8" intra-aortic balloon pumping catheter for preventing limb lschemia. AB - The most important limitation in the use of an intra-aortic balloon pumping (IABP) is the risk of vascular complications. Recently, an IABP catheter with an 8.0 French shaft and low profile that may decrease the risk of vascular morbidity has been developed. We evaluated the in vitro balloon performance and the prevention of limb ischemia in clinical use. An 8.0 French IABP catheter was compared with a standard 9.5 French catheter. Inflation time (IT), deflation time (DT), and changes in volume (V) generated by the balloon were measured during the pumping cycle in an experimental model. The inflation velocity (V/IT) and deflation velocity (V/DT) were calculated as parameters of balloon performance. At 120 bpm the V/IT and V/DT were 0.34 ml/msec and 0.28 ml/msec with the 8.0 French, and 0.33 ml/msec and 0.24 ml/msec with the 9.5 French catheter. Twelve patients with coronary artery disease, ranging in age from 41 to 87 years (mean, 66 years), who underwent IABP support, were divided into group 1 (8.0 French, n = 4) and group 2 (9.5 French, n = 8). Ankle-arm pressure index (API), lactate extraction ratio (LER) in the limb with IABP insertion, and cardiac index (CI) were measured at 1, 12, and 24 hours postoperatively. There were no major vascular complications and no counterpulsation related morbidity. There was no significant difference between the two groups with regard to age, duration of IABP support, and incidence of peripheral vascular disease and diabetes. The percentage of women patients was significantly higher in group 1 (100% vs. 25%), whereas body surface area was significantly smaller (1.45 +/- 0.14 vs. 1.68 +/- 0.12 m2). The API in group 1 were slightly higher than those in group 2 throughout the observed period (not significant). The LER and Cl showed no significant differences between the two groups. These results suggest that the 8.0 French IABP catheter with a low profile has an acceptable in vitro performance, and its clinical application may be effective in preventing limb ischemia in a high-risk subset of patients such as women and smaller patients. PMID- 11730198 TI - Preserving central blood volume: changes in body fluid compartments during hemodialysis. AB - The understanding of fluid changes during hemodialysis (HD is essential for reducing complications as well as efficacy of the procedure. Bioimpedance spectroscopy provides a non invasive method of measuring total body water (TBW), the distribution of intra (ICF) and extracellular (ECF) fluids, and their changes during HD. Segmental bioimpedance may be used to measure the same fluid shifts but from different body segments; the technique has previously been shown to com pare well with whole body measures. It is possible that fluid shifts occur differently in different body compartments during HD. Based on previous hemodynamic studies we postulated that during HD ultrafiltration (UF) the body attempts to preserve its central blood volume (cardiopulmonary circula tion plus great vessels), and thus fluid shifts would be greater from the periphery than from central compartments. To test this hypothesis, segmental bioimpedance (Xitron Technolo gies, San Diego, CA) was performed on 11 subjects undergoing HD where ECF and ICF values were obtained from the legs, arms and trunk before and after a period of UF. Blood volume change (ABV%) was also followed using an on line optical hematocrit (Hct) sensor (Crit-Line monitor, In-Line Diagnostics, UT) where deltaBV% = deltaBV% = (1 - Hct1/Hct0) x 100 (Hct0 = baseline Hct; Hct1 = postultrafiltration Hct) The UF of 2.0 L +/- 0.79 L (M +/- SD) over 75 minutes was associated with a deltaBV% of -9.43% +/- 3.6% (M +/- SD), a significant (Student's paired t-test) reduction in total body (TB) ECF (p < 0.02), a weak correlation in reduction in TBW (p = 0.09) but not in TB ICF. The ECF reductions from the trunk, legs, and arms were all significant (minimum p < 0.02); no ICF changes from these compartments were significant. The amount of ECF reduction was greater from the legs (0.7 L +/- 0.6 L) than the arms (0.12 L +/- 0.08 L) and trunk (0.2 L +/- 0.2 L) (all M +/- SD). Multiple regression analysis showed that TB ECF changes correlated strongly with leg (r = 0.94, p < 0.001) and arm (r = 0.72, p = 0.002) ECF changes but not with trunk changes. deltaBV% correlated weakly with leg (r = 0.45, p = 0.08) and arm (r = 0.42, p = 0.10) ECF changes but not with the trunk. As the deltaBV% represents the net volume change between UF and plasma water refilling, thiss indicates that plasma water is being removed more from the peripheral compartments than from the trunk. These data suggest that plasma refilling during HD to preserve central blood volume is more dynamic from the leg ECF than from elsewhere and may, in turn, explain the frequent occurrence of leg cramps during and after hemodialysis. PMID- 11730199 TI - Effect of vitamin E modified cellulose membrane on human lymphocyte, monocyte, and granulocyte CD11b/CD18 adhesion molecule expression during hemodialysis. AB - Chronic renal failure induces a clinical state of immunodefi ciency that also depends upon a wide spectrum of dialysis membranes used during hemodialysis. Previous studies have shown that cellular immunodeficiency is caused by malfunc tion of the antigen presenting cells (monocytes or granulocytes). Subsequent activation of rolling mononuclear leuko cytes results in up-regulated expression of CD11b/CD18 (Mac-1) on endothelial cells. It is postulated that a VitE coated dialysis membrane might minimize the membrane biocompatibility, thereby generating a smaller amount of re active oxygen species (ROS). The purpose of this study was to evaluate the expression of the CD11b/CD18 adhesion mole cule on lymphocytes, monocytes, and granulocytes during HD in 10 patients, using flow cytometric analysis. The study protocol included the measurement of molecule expression using cellulose membrane (Clirans RS15, TERUMO Corp. Japan), and the same membrane coated by vitamin E (Exce brane, Clirans E15, TERUMO Corp., Japan) during 20 dialysi sessions each. Lymphocyte CD11 b/CD1 8 (Mac-1) expression did not change with either dialyzer type. However, monocyt (p = 0.046) and granulocyte (p = 0.018) CD11b/CD18 ex pression in the post HD period was significantly lower using the vitamin E coated membrane compared with the contro cellulose membrane. Our findings suggest a significant de crease in activation and migration of monocytes and granu locytes when using a vitamin E coated cellulose membrane. PMID- 11730200 TI - Determination of extra renal clearance and generation rate of beta2-microglobulin in hemodialysis patients using a kinetic model. AB - Beta-2-microglobulin (beta2-m) has been proposed as a marker of middle molecules to assess one aspect of the efficacy of dialysis. Until now, few data have been published about extra renal (metabolic) clearance and generation rates of beta2 microglobulin, which are necessary for calculation of total clearance and mass removal of beta2-m in hemodialysis patients. We have developed a simple method to derive extra renal clearance and generation rates of beta2-m by measuring the pre and post dialysis blood concentrations of beta2-m using kinetic modeling. Ten stable hemodialysis patients were included in this study. Pre and post dialysis concentrations of beta2-m were measured during dialysis with low flux dialyzers (F6 HPS) and after 10 days switching to high flux dialyzers (F60S or Superflux). With a validated two pool model, the generation rate of beta2-m can be determined if extra renal clearance is known. Assuming the generation rate of beta2-m to be constant in each patient, the computer reiterated the calculation of extra renal clearance until the calculated generation rate was equal for both the low flux and the high flux dialyzer. Extra renal clearance was found to be between 1.97 and 4.11 ml/min (average, 3.2 ml/min). Generation rate was found in a rather narrow range between 1.63 and 2.56 mg/kg per day (average, 2.09 mg/kg per day). There was no correlation between extra renal clearance and generation rates. With this simple method, extra renal clearance and generation rates of beta2-m can be determined using data by switching hemodialysis patients from impermeable to permeable membranes. PMID- 11730201 TI - Mass transfer characteristics of artificial lungs. AB - An artificial lung is used during cardiopulmonary bypass to oxygenate blood and control blood temperature. The oxygen transfer rate-flow rate characteristics of three hollow fiber membrane artificial lungs (Sarns Turbo 440, Cobe Optima, Dideco Compactflo) were determined in vitro to characterize design features. Results are presented as a unique dimensionless relationship between Sherwood number, NSh (ratio of convective to diffusive mass transfer), Schmidt number, NSc (ratio of momentum to diffusive transport), and Reynolds number, NRe (ratio of inertial to viscous forces). This relationship is a function of device porosity, epsilon, and characteristic device length, xi, defined as the ratio of the mean blood path and manifold length: Nsh/NSc(1/3) x xi(1/2) = phi x (epsilon(1/m) x NRe)(m) where phi = 0.26 and m = 1.00 for NPe < 3,200 and phi = 0.47 and m = 0.64 for NPe > 3,200 where NPe is the dimensionless Peclet number defined as NRe x NSc. We found good correspondence between the model predictions and in vitro blood oxygen transfer rates. We conclude that this dimensionless approach allows us (1) to compare artificial lungs independently, (2) to relate water tests to blood, and (3) to predict the oxygen transfer rate of a new artificial lung design. PMID- 11730202 TI - Development of infection resistant polyurethane biomaterials using textile dyeing technology. AB - Infection is a major complication when using biomaterials such as polyurethane in the clinical setting. The purpose of this study was to develop a novel infection resistant polyurethane biomaterial using textile dyeing technology. This procedure results in incorporation of the antibiotic into the polymer, resulting in a slow, sustained release of antibiotic from the material over time, without the use of exogenous binder agents. Polycarbonate based urethanes were synthesized that contained either a non-ionic (bdPU) or anionic (cPU) chain extender within the polymer backbone and cast into films. The fluoroquinolone antibiotic ciprofloxacin (Cipro) was applied to bdPU and cPU using textile dyeing technology, with Cipro uptake determined by absorbance reduction of the "dyebath." These dyed bdPU/cPU samples were then evaluated for prolonged Cipro release and antimicrobial activity by means of spectrophotometric and zone of inhibition assays, respectively. Cipro release and antimicrobial activity by dyed cPU segments that were aggressively washed persisted over 9 days, compared with dyed bdPU and dipped cPU control segments that lasted < 24 hours. Dyed cPU segments, which remained in a static wash solution, maintained antimicrobial activity for 11 days (length of study), whereas controls again lost antimicrobial activity within 24 hours. Thus, application of Cipro to the cPU polymer by means of dyeing technology results in a slow sustained release of antibiotic with persistent bacteriocidal properties over extended periods of time. PMID- 11730203 TI - A gelatin coated collagen-polyglycolic acid composite membrane as a dural substitute. AB - We designed a new biodegradable composite membrane for use as a dural substitute. This membrane is composed of polyglycolic acid (PGA) mesh, collagen sponge, and gelatin sponge. Collagen layers are attached on both sides of the PGA mesh. One side of the membrane is coated with freeze-dried gelatin to prevent tissue adhesion. We examined the mechanical properties of the membrane and evaluated its efficacy in vivo by implanting it into dogs. When the mechanical properties of the membrane were measured under wet conditions, tensile strength and resistance against cutting by a suture were approximately 20 N/cm and 13 N, respectively, which are adequate for a surgical material. In the in vivo experiment, our dural substitute was implanted into six adult beagle dogs to repair a 25 x 15 mm defect in the dura mater. Two dogs each were killed at 2 and 4 months after implantation and two were observed over 6 months. At 4 months after implantation, the implanted dural substitute was almost absorbed and a fibrous membrane resembling native host dura had regenerated at the site. No foreign body reaction or significant adhesion to the cerebral cortex was observed. These results indicate the clinical potential of our novel dural substitute. PMID- 11730204 TI - New approach to radial expansive force measurement of self expandable esophageal metal stents. AB - The accurate measurement of radial expansive force is crucial for optimal design and implantation of self expandable esophageal metal stents. In the present study, a new method of measurement under experimental conditions simulating actual stent implantation has been developed. This method offers precise and reproducible measurements and can be applied to a wide variety of stent types. In particular, the method enables one to measure expansive pressure as well as the true radial expansive force up to the radial compression ratio of 72%, covering the range of compression often encountered in a partially obstructed lumen. The test results for various kinds of metal stents are presented and compared. Based on these results, three important points of observation critical in explaining and predicting the expansion characteristics of stents have been reported. Further understanding and characterization of these findings will be necessary for developing new stents with outstanding clinical efficacy. PMID- 11730205 TI - Vacuum assisted venous drainage does not increase trauma to blood cells. AB - Although gravity drainage has been the standard technique for cardiopulmonary bypass (CPB), the development of min imally invasive techniques for cardiac surgery has renewed interest in using vacuum assisted venous drainage (VAVD) Dideco (Mirandola, Italy) has modified the D903 Avant oxygenator to apply a vacuum to its venous reservoir. The impact of VAVD on blood damage with this device is analyzed. Six calves (mean body weight, 71.3 +/- 4.1 kg) were con nected to CPB by jugular venous and carotid arterial cannu lation, with a flow rate of 4-4.51 L/min for 6 h. They were assigned to gravity drainage (standard D903 Avant oxygen ator, n = 3) or VAVD (modified D903 Avant oxygenator, n = 3). The animals were allowed to survive for 7 days. A standard battery of blood samples was taken before bypass, throughout bypass, and 24 h, 48 h, and 7 days after bypass. Analysis of variance was used for repeated measurements. Thrombocyte and white blood cell counts, corrected by hematocrit and normalized by prebypass values, were not significantly different between groups throughout all study periods. The same holds true for hemolytic parameters (lactate dehydrogenase [LDH] and plasma hemoglobin). Both peaked at 24 hr in the standard and VAVD groups: LDH, 2,845 +/- 974 IU/L vs. 2,537 +/- 476 IU/L (p = 0.65), respectively; and plasma hemoglobin, 115 +/- 31 mg/L vs. 89 +/- 455 mg/L (p = 0.45), respectively. In this experimental setup with prolonged perfusion time, VAVD does not increase trauma to blood cells in comparison with standard gravity drainage. PMID- 11730206 TI - Can latissimus dorsi muscle stimulation benefit heart during training period after vascular delay? AB - We hypothesized that a two-stage vascular delay procedure followed by 5 weeks of conditioning of the latissimus dorsi muscle (LDM) could benefit the heart during the training period and greatly increase cardiac assistance when examined with maximum potential. In mongrel dogs (n = 10), left ventricle (LV) dysfunction was induced by intracoronary injections of latex microspheres [90 +/- 2 micro diameter]. Vascular delay of the LDM was performed in one group (n = 6), whereas the other group (control, n = 4) did not undergo vascular delay. After 2 weeks, CMP was performed in all animals followed by LDM conditioning. After 5 weeks of muscle training, we examined left ventricular function at 20 Hz-4 volts, 33 Hz-4 volts, and 50 Hz-10 volts stimulation by assessing peak aortic pressure (AoP), left ventricular pressure (LVP), maximum LV +dP/dt, stroke volume (SV), stroke work (SW), stroke power (SP), and aortic flow. LDM assisted beats were compared with nonstimulated beats. LDM stimulation caused significant increases in pressure and flow in the vascular delay group. At 20 Hz-4 volts, absolute increases were LVP (10.2 +/- 0.6) mm Hg, AoP (9.8 +/- 1.7) mm Hg, SV (1.8 +/- 0.4) ml, SW (5.3 +/- 1.0) gm x m, SP (40.8 +/- 12.7) gm x m/sec, max LV dP/dt (104.8 +/- 53.2) mm Hg/sec, and peak aortic flow (0.9 +/- 0.3) L/min. At 33 Hz-4 volts, the absolute increases were LVP (13.6 +/- 1.3) mm Hg, AoP (12.1 +/- 2.4) mm Hg, SV (2.7 +/- 0.7) ml, SW (7.4 +/- 1.4) gm x m, SP (72.7 +/- 16.5) gm x m/sec, max LV dP/dt (294 +/- 19) mm Hg/sec, and peak aortic flow (1.8 +/- 0.5) L/min. At 50 Hz-10 volts, the absolute increases were LVP (17.7 +/- 0.7) mm Hg, AoP (21.1 +/- 1.9) mm Hg, SV (6.0 +/- 1.1) ml, SW (14.6 +/- 2.2) gm.m, SP (128.2 +/- 15.3) gm x m/sec, max LV dP/dt (352 +/- 62) mm Hg/sec, and peak aortic flow (3.3 +/- 0.4) l/min (p < 0.05). The percentage increases were significantly larger in the vascular delay group compared with controls at 50 Hz-10 volts LDM stimulation. By using a two-stage vascular delay procedure, LDM stimulation can provide meaningful cardiac assistance during training periods. Furthermore, brief periods of maximal potential benefit (demand cardiomyoplasty) can be achieved during the training period. PMID- 11730208 TI - Chronic heart failure model with sequential ligation of the homonymous artery and its diagonal branch in the sheep. AB - We report a reliable chronic heart failure model in sheep using sequential ligation of the homonymous artery and its diagonal branch. After a left anterior thoracotomy in Corridale sheep, the homonymous artery was ligated at a point approximately 40% of the distance from the apex to the base of the heart, and after 1 hour, the diagonal vessel was ligated at a point at the same level. Hemodynamic measurements were done preligation, 30 minutes after the homonymous artery ligation, and 1 hour after diagonal branch ligation. The electrocardiograms were obtained as needed, and cardiac function was also evaluated with ultrasonography. After a predetermined interval (2 months for five animals and 3 months for two animals), the animals were reevaluated in the same way as before, and were killed for postmortem examination of their hearts. All seven animals survived the experimental procedures. Statistically significant decreases in systemic arterial blood pressure and cardiac output and increases in pulmonary artery capillary wedge pressure were observed 1 hour after sequential ligation of the homonymous artery and its diagonal branch. Untrasonographic analyses demonstrated variable degrees of anteroseptal dyskinesia and akinesia in all animals. The data from animals at 2 months after coronary artery ligation showed significant increases in central venous pressure, pulmonary artery pressure, and pulmonary artery capillary wedge pressure. Left ventricular enddiastolic dimension and left ventricular end-systolic dimension on ultrasonographic studies were also increased. Electrocardiography showed severe ST elevation immediately after the ligation and pathologic Q waves were found at 2 months after ligation. The thin walled infarcted areas with chamber enlargement were clearly seen in the hearts removed at 2 and 3 months after ligation. In conclusion, we could achieve a reliable ovine model of chronic heart failure using a simple concept of sequential ligation of the homonymous and diagonal arteries. This animal model was comparable to the clinical situation. PMID- 11730207 TI - Is an endocardial connection necessary for growth factor induced angiogenesis in transmyocardial laser revascularization? AB - Transmyocardial laser revascularization (TMLR) and therapeutic angiogenesis had emerged as potential tools in the treatment of angina refractory to conventional therapies. This combination might potentiate their effects, because angiogenesis is believed to be a basic mechanism in TMLR. The influence of channel connection with endocardial blood flow on angiogenesis is unclear. Twenty-five pigs (mean weight, 72.3 +/- 5 kg) were randomly assigned into five groups. In the transmural laser group, five transmyocardial channels were drilled. In the transmural mixed group, the same protocol was used followed by the injection of 100 microg of bovine bone derived growth factor mixture within each channel. The nontransmural laser group and the nontransmural mixed group underwent the same procedures, respectively, but the laser channels were drilled through the outer two-thirds of the myocardial wall. The control group had sham operations. Animals were allowed to survive for 1 month. Vascular densities were determined by computed morphometric analysis of histologic sections. Vascular counts of areas adjacent to the channels in the non- and transmural laser groups did not differ significantly from control groups (arteriolar counts: 0.27 +/- 0.16 and 0.26 +/- 0.16 vs. 0.29 +/- 0.11/mm2, respectively). When bovine bone protein growth factor mixture is added, neovascularization is increased significantly in non- and transmural mixed groups (1.04 +/- 0.79 and 0.69 +/- 0.37/mm2, respectively, p < 0.001 for both comparisons with corresponding laser groups), and there was no significant difference between mixed groups (p = 0.13). In this porcine model, the combination of TMLR with injection of bone protein growth factor mixture induced angiogenesis around the laser channels. Whether the channels did or did not communicate with the endocardial cavity did not influence the neovascular density. PMID- 11730209 TI - Prostaglandin synthesis inhibitor improves hypotension during normothermic cardiopulmonary bypass. AB - Hypotension is a major systemic side effect during cardiopu monary bypass (CPB), especially at normothermia. We previously reported that prostaglandin (PG) might play a substantial role in hypotension. The purpose of this study was to clarify whether a PG synthesis inhibitor (PGSI) could improve hypotension during CPB. Thirty-six patients undergoing cardiac surgery with normothermic CPB (35-37 degrees C) were divided into two groups: a PGSI group (n = 18), whose members wer given a PGSI before and during CPB, and a control group (n = 18). In both groups, perfusion flow was sufficient and pressure was maintained at above 45 mm Hg by infusion of metaraminol, a vasoconstrictor. The mean arterial pressure throughout CPB was significantly higher in the PGSI group than in the control group (57 +/- 4 vs. 48 +/- 3 mm Hg, p < 0.01), whereas the dose of infused metaraminol was significantly lower in the PGSI group (13 +/- 7 vs. 21 +/- 6 mg, p < 0.01). The blood base excess was not significantly different (1.0 +/- 1.6 vs. 1.7 +/- 1.9 mmol/L, p = 0.28), and urine output was significantly higher in the PGSI group (503 +/- 179 vs. 354 +/- 112 ml/hr, p < 0.01). In conclusion, PGSI can improve hypotension during CPB and increase urine output without impairing peripheral circulation. PMID- 11730210 TI - Fourteen year experience with the omnicarbon prosthetic heart valve. AB - The Omnicarbon prosthetic valve was implanted in 168 patients (mean age, 53 years); 84 had aortic valve replacement (AVR), 57 had mitral valve replacement (MVR), and 27 had both aortic and mitral valve replacement (DVR). The mean follow up period was 6.8 years, with a maximum of 13.7 years. Three patients (1.8%) were lost to follow-up. There were 4 (2.4%) early deaths and 26 late deaths. Survival at 10 years was 76% in the AVR and MVR groups, and 85% in the DVR group. Freedom from thromboembolism at 10 years was 94% in the AVR group, 80% in the MVR group, and 92% in the DVR group. Freedom from hemorrhagic complications at 10 years was 86% in the AVR group and 92% in the MVR group. At 10 years, 97% in the AVR group and 96% in the MVR group were free from endocarditis. One patient in the DVR group suffered a paravalvular leak. At 10 years, 97% in the AVR group and 95% in the MVR group had not needed reoperation. Elevation of the postoperative serum lactate dehydrogenase levels were acceptable in the three groups. In conclusion, the Omnicarbon prosthetic valve has shown excellent long-term clinical results. PMID- 11730211 TI - Blood lactic acid levels after artificial heart implantation. AB - The aim of this study is to investigate blood lactic acid levels after the placement of the AbioCor implantable replacement heart to determine whether circulatory support with the AbioCor device results in adequate tissue perfusion. Tissue perfusion may not be adequate during cardiac surgery, especially with the use of cardiopulmonary bypass (CPB). Inadequate perfusion is usually associated with anaerobic metabolism, metabolic acidosis, and lactate accumulation. Ten calves had implantation of the AbioCor by means of a right thoracotomy. Standard cardiopulmonary bypass was performed during surgery. Lactic acid levels and blood gas analyses were measured. All animals were monitored for pressures in the aorta, pulmonary artery, and left and right atria. The output of the AbioCor was calculated based on beat rate and stroke volume. All animals were rapidly weaned off cardiopulmonary bypass to full AbioCor support and had normal hemodynamics with normal filling pressures. The lactic acid levels were elevated after surgery, peaked at 8 to 12 hours, and then gradually decreased to the normal range 24 hours after surgery. There was a positive relationship between lactate levels, oxygen consumption, and extraction rate, that is, the lactic acid levels were higher in the presence of increased oxygen consumption and increased oxygen extraction rate. There was a negative relationship between lactate levels and oxygen delivery, and arterial and venous oxygen content, that is, the lactic acid levels were higher when oxygen delivery and arterial and venous oxygen contents were low. The pathophysiology of lactic acidosis is uncertain, but is most likely due to inadequate perfusion during cardiopulmonary bypass, and lactate washout after adequate perfusion has been established. The return to aerobic metabolism with clearance of lactate demonstrates that the AbioCor is able to provide complete circulatory support with normal tissue perfusion. PMID- 11730212 TI - Automatic monitoring system for artificial hearts using self organizing map. AB - This study presents an automatic monitoring system for artificial hearts. The self organizing map (SOM) was applied to monitoring and analysis of an aortic pressure (AoP) signal measured from an adult goat equipped with a total artificial heart. In the proposed system, two different SOMs were used to detect and classify abnormalities in the measured AoP signal. In the first stage, an ordinary SOM, taught with only normal AoP data, was used for detection of abnormalities on the basis of the quantization error in the real-time monitoring task. In the second stage, a supervised SOM was used for classification of abnormalities. The supervised SOM can be regarded as an ordinary SOM with an extra class vector for solving the classification problem. The class vector is assigned to every node in the second SOM as an output weight learned according to Kohonen's learning rule. The effectiveness of detection and classification of abnormalities using these two SOMs was confirmed. PMID- 11730213 TI - Efficiency of an air filter at the drainage site in a closed circuit with a centrifugal blood pump: an in vitro study. AB - In a closed circuit with a centrifugal blood pump, one of the serious obstacles to clinical application is sucking of air bubbles into the drainage circuit. The goal of this study was to investigate the efficiency of an air filter at the drainage site. We used whole bovine blood and the experimental circuit consisted of a drainage circuit, two air filters, a centrifugal blood pump, a membrane oxygenator, a return circuit, and a reservoir. Air was injected into the drainage circuit with a roller pump, and the number and size of air bubbles were measured. The air filter at the drainage site could remove the air bubbles (>40 microm) by itself, but adding a vacuum removed more bubbles (>40 microm) than without vacuum. Our results suggest that an air filter at the drainage site could effectively remove air bubbles, and that adding the filter in a closed circuit with a centrifugal blood pump would be safer. PMID- 11730214 TI - Evaluation of a patient with left ventricular assist device dysfunction. AB - With the increasing number of LVAD recipients due to lack of donor availability and use of an LVAD as destination therapy, there will be a significant proportion of this population presenting with device related complications. Initial evaluation should be with TEE because conduit obstruction would be a significant complication, among others. As observed in this case report, it is sometimes difficult to noninvasively detect device malfunction, and even surgical exploration left questions unanswered. Echocardiography has been the principal modality used so far to assess the LVAD cannulas. More reliance on Doppler velocities and establishment of a normal range of values and flow profiles of the inflow and outflow cannulas are necessary. Baseline echocardiographic studies could be used for future comparisons. Fluoroscopy and selective angiography have also been reported to be helpful. We would also recommend measuring LV hemodynamics, which have not been previously described, as an additional mode of evaluation of this emerging problem. This case report demonstrates the need for additional noninvasive modalities for diagnosis of ventricular assist device malfunction. PMID- 11730215 TI - Did Undar and associates really make those comments on the issue of pulsatile versus nonpulsatile flow? Absolutely not. PMID- 11730216 TI - Influence of preoperative computed tomography on patients undergoing appendectomy. AB - The frequency of computed tomography (CT) ordered by emergency department physicians at our facility was noted to sharply increase in early 1998 after a New England Journal of Medicine (NEJM) article recommending routine CT in patients with suspected appendicitis. Numerous studies have proven the accuracy of CT for detecting acute appendicitis; however, the most appropriate use of CT continues to evolve. We sought to evaluate the effect of increased CT use on negative appendectomy rate and perforation rate at our institution and to better delineate in whom CT is most beneficial. CT use was retrospectively evaluated and found to sharply increase in April 1998. The authors then reviewed the medical records of 291 consecutive patients undergoing appendectomy 18 months before and after the NEJM article. Patients with interval appendectomies and those 12 years of age or younger were excluded. The remaining 226 patients constitute the study cohort. The study cohort was then divided into the two groups. The "Discriminate Group" consists of patients from the 18 months before the NEJM article impact and a period of selective CT use. The "Indiscriminate Group" comprises patients from the subsequent 18 months in which CT use was substantially higher and routinely obtained before surgical evaluation. After chart review an objective clinical score (Alvarado score) was assigned to each patient. Comparison was then made between the two groups on perforation rate, negative appendectomy rate, time delay to operating room, and Alvarado score. Additionally patients undergoing preoperative CT were compared with those without CT. These groups were also evaluated on the basis of negative appendectomy rate, perforation rate, and delay to the operating room. CT in patients with abdominal symptoms associated with appendicitis increased from 188 in the Discriminate Group to 1035 in the Indiscriminate Group. In the Discriminate Group the negative appendectomy rate was 15.1 per cent. After the indiscriminate use of CT the negative appendectomy rate decreased to 13.3 per cent, but this was not significant. Males experienced a decrease in the negative appendectomy rate from 10.1 to 6.9 per cent, whereas the rate for females increased slightly from 21.3 to 22.9 per cent. Again we found no statistical significance in these changes. The overall perforation rate of 17.9 per cent in the first 18 months decreased to 13.3 per cent in the following 18 months but again was not statistically significant. The Alvarado scores between the Discriminate and Indiscriminate groups were 6.7 and 7.3, respectively (P = 0.02). Patients with preoperative CT averaged 11.9 hours to the operating room compared with 6.5 hours for those without CT (P = 0.03). Use of CT did not decrease perforation rate but did globally reduce negative exploration (P = 0.05). This reduction in negative exploration however was not discriminated by sex. CT use in suspected acute appendicitis has greatly increased over the past several years. The dramatic increase in CT use at our institution has not resulted in dramatic decreases in negative appendectomy rate or statistically significant changes in perforation rate. The optimal use of CT in evaluating patients with suspected appendicitis has yet to be determined. Surgical consultation should be obtained early to avoid indiscriminate tests. PMID- 11730217 TI - Minimally invasive parathyroidectomy without intraoperative localization. AB - Minimally invasive parathyroidectomy (MIP) is gaining popularity as an alternative to traditional bilateral exploration for patients with primary hyperparathyroidism. The success of MIP relies on the ability of preoperative and intraoperative localization studies to guide a directed exploration for resection of a diseased gland. We hypothesize that excellent results can be achieved with MIP when only technetium-99m sestamibi (MIBI) is used for localization. We conducted a prospective analysis of all patients presenting with a biochemical diagnosis of primary hyperparathyroidism between January 1997 and November 2000. Patients meeting inclusion criteria were given a choice of MIP and directed exploration versus traditional bilateral exploration. Fifty patients chose MIP. Three patients who chose MIP had a negative MIBI, which left 47 patients in the primary study group. The MIBI correctly identified a parathyroid adenoma in 42 patients (89.3%). In two other patients MIBI was inaccurate; however, directed exploration was successfully converted to a bilateral exploration. Overall 44 of 47 (93.6%) patients in the study group were rendered normocalcemic after the initial operation. Three patients experienced persistent hypercalcemia and subsequently underwent successful bilateral exploration. Including those patients choosing a bilateral exploration, a total of 59 positive MIBI scans were evaluated. There were 54 true positives (positive predictive value 91.5%), and if all patients had chosen a MIP 94.9 per cent would have been successfully treated at the initial operation. Mean operative time for MIP was 54.6 minutes, and in 32 patients (68.1%) MIP was performed with local anesthesia and sedation. Twenty-six patients (55.3%) were discharged the same day of the procedure. There were no significant complications in any group analyzed. We conclude that MIP can be successfully performed on the basis of a positive MIBI scan. The present study highlighting many of the advantages of MIP questions the necessity of additional adjuncts such as intraoperative parathyroid hormone measurement and gamma-probe localization. PMID- 11730218 TI - Institutional learning curve for sentinel node biopsy at a community teaching hospital. AB - Sentinel lymph node biopsy (SLB) is gaining popularity as an alternative to axillary lymph node dissection for breast cancer staging. Although publications have described the inherent learning curve few have analyzed actual performance in community hospitals. This study analyzes the institutional learning curve for SLB in a community teaching hospital without a formal sentinel node credentialing policy. We conducted an analysis of the initial 96 SLBs performed by 15 general surgeons over a 34-month period. The main outcomes were rate of identification of sentinel node and accuracy of SLB. Overall SLB was successful in identifying one or more sentinel nodes (mean = 2.2) in 73 per cent of attempted cases. There were marked differences in performance of individual surgeons; identification rates varied from 25 to 100 per cent. Only one surgeon performed more than 15 procedures during the study period. Nineteen of 21 cases with positive nodes were correctly characterized (sensitivity = 90.5%; 95% confidence interval = 76-100%; false negative rate = 9.5%). Our institutional learning curve was longer than high-volume individual experiences published in the literature, with a lower rate of sentinel node identification. SLB appears to be sensitive for detecting malignancy, but the small number of patients with positive nodes in our series limits our conclusions. The marked variability in individual surgeon performances and the slow rate of overall improvement in our institution suggest a need for a formalized policy for SLB training. PMID- 11730219 TI - Operative acute small bowel obstruction: admitting service impacts outcome. AB - Early surgical intervention in acute small bowel obstruction (SBO) has long been recognized as an important factor in preventing morbidity and mortality. Factors associated with surgically managed acute SBO were analyzed for delay in intervention and impact on outcome. A retrospective review of all patients evaluated for SBO on the surgical teaching service of the Greenville Hospital System from July 1, 1997 to June 30, 2000 was performed. Data were collected on patient demographics, admission information (date, admitting service, physical examination, and laboratory values), comorbidity, diagnostic studies, surgery date, operative findings, postoperative complications, operative mortality, and discharge date. Analysis of the data revealed 157 cases of presumed SBO of which 61 were managed nonoperatively and 96 required surgery. Acute SBO was diagnosed in 65 patients who constitute the basis for this review. Of these 65 patients 43 (66%) were admitted to the surgical service, 25 (38%) required small bowel resection, and 17 (26%) developed morbidity and/or mortality. When analyzed for morbidity and mortality the only characteristics that were statistically significant were the admitting service (P = 0.003) and length of stay (P = 0.003). On further analysis of admitting service and patient outcomes several factors were significant when we compared medical service admissions to surgical service admissions. These included days from admission to surgery (P = 0.003), length of stay (P = 0.019), morbidity (P = 0.004), mortality (P = 0.005), and combined morbidity and mortality (P = 0.003). Mortality of patients admitted to the medical service was 27 per cent compared with 2 per cent for the surgical service. There were no differences in morbidity and mortality when analyzed by the need for small bowel resection, patient age, etiology of obstruction, or presence of comorbidities. None of the factors studied were useful in predicting the need for small bowel resection. Our findings agree with those of previous investigators with regard to 1) lack of association between the preoperative evaluation and the need for small bowel resection and 2) the association between delay in diagnosis and increased morbidity and mortality. In addition we have found that one of the primary causes of delay in treatment for SBO was admission to the medical service. This delay led to significantly higher mortality in these patients. We recommend early surgical evaluation for any patient admitted with SBO as a differential diagnosis. PMID- 11730220 TI - Radical resection improves survival for patients with pT2 gallbladder carcinoma. AB - Radical resection (wedge resection of the gallbladder bed and dissection of the hepatoduodenal ligament, portal, and celiac lymph nodes) has been reported to improve survival from pathologic T2 gallbladder carcinoma (pT2 GBCa; invasion through the muscularis without perforation of the serosa). We report our experience and the outcome of patients with pT2 GBCa. Between 1989 and 2000 at Vanderbilt University Medical Center ten patients were found to have pT2 disease after cholecystectomy. The patients had an average age of 64+/-13 years and underwent either radical resection (n = 5) or no further surgical therapy (n = 5). Of the patients who underwent cholecystectomy only, one (20%) is still alive at 27 months and four (80%) died of recurrent GBCa between 6.5 and 21 months. For the patients who underwent radical resection all five are alive at 15 to 83 months with no recurrence. The proportion of patients surviving pT2 GBCa after radical resection was significantly greater than with cholecystectomy alone (P < 0.05). The difference in length of survival between the two groups was also significant (P < 0.05). Morbidity after radical resection was low (pancreatic leak in one patient), and there were no operative mortalities. Radical resection significantly improved survival over cholecystectomy alone for patients with pT2 GBCa. The procedure has low morbidity and mortality rates. Therefore a radical resection operation is indicated for patients with pT2 GBCa. PMID- 11730221 TI - Anal carcinoma or "just hemorrhoids"? AB - Cancers of the anal margin and anal canal are extremely rare and often misdiagnosed. Only one to two per cent of large bowel cancers arise in this area. Current management of these cancers includes surgery, combined chemoradiation, or both. From January 1985 through July 2000, 50 patients were diagnosed with anal cancer at two institutions. This retrospective review includes all available cases of anal cancer including all histologies. Patient charts were analyzed for diagnosis, staging, treatment, survival, and recurrence rate. The patients ranged in age from 27 to 92 years (median age 51 years; mean age 52.8 years); there were 22 men and 28 women. The pathologic diagnosis included 44 (88%) with squamous cell carcinoma, three (6%) with melanoma, two (4%) with adenocarcinoma, and one (2%) with Paget's disease. At presentation nine (18%) were classified as stage 0, five (10%) stage I, 21 (42%) stage II, eight (16%) stage III, and seven (14%) stage IV. Mean follow-up data were available on 100 per cent of the patients. Chemoradiotherapy was the primary treatment modality in 25 patients (50%). Ten patients (20%) underwent abdominoperineal resection (APR) in the study. Three patients (6%) received an APR as primary treatment, three (6%) in combination with chemoradiation, and four (8%) for salvage therapy. Fourteen patients (28%) underwent wide local excision (WLE) as the primary treatment. Two patients (4%) underwent WLE plus chemoradiation therapy. One patient (2%) underwent WLE and chemotherapy. There were 18 deaths (36%) in this series. Thirteen patients (26%) died of anal cancer; the average time to death from diagnosis was 13.2 months. Three of these deaths were in patients with melanoma who presented with stage IV disease. Thirty-two patients (64%) are alive, and 30 (60%) of these patients are free of disease (mean time since diagnosis 32.5 months, range 2-151 months). Six patients (12%) had recurrence after treatment (mean time to recurrence 12.6 months; range 3-26 months). Anal cancers continue to present at an advanced stage, with a high mortality rate. Anal melanoma in particular is an aggressive and highly fatal cancer. APR remains the recommended salvage therapy for advanced anal carcinomas that fail primary treatment. In our series only one of four patients has had a disease-free survival of 4 months. Early recognition and detection of primary and recurrent disease is necessary for improved outcome. PMID- 11730222 TI - Laparoscopic anterior esophageal myotomy and toupet fundoplication for achalasia. AB - Achalasia is an esophageal motility disorder characterized by the failure of lower esophageal sphincter relaxation and the absence of esophageal peristalsis. The purpose of this study was to evaluate the clinical outcomes of patients undergoing laparoscopic esophageal myotomy and Toupet fundoplication for achalasia. A 9-cm myotomy was performed in most cases extending 7 cm above and 2 cm below the gastroesophageal junction. Severity of dysphagia, heartburn, chest pain, and regurgitation was graded preoperatively and postoperatively using a five-point symptomatic scale (0-4). Patients also graded their outcomes as excellent, good, fair, or poor. Between December 1995 and November 2000 a total of 49 patients (23 male, 26 female) with a mean age of 44.3 years (range 23-71 years) were diagnosed with achalasia. Mean duration of symptoms was 40.2 months (range 4-240 months). Thirty-seven patients (76%) had had a previous nonsurgical intervention or combinations of nonsurgical interventions [pneumatic dilation (23), bougie dilation (five), and botulinum toxin (19)], and two patients had failed esophageal myotomies. Forty-five patients underwent laparoscopic esophageal myotomy and Toupet fundoplication. Two patients received laparoscopic esophageal myotomies without an antireflux procedure, and two were converted to open surgery. One patient presented 10 hours after a pneumatically induced perforation and underwent a successful laparoscopic esophageal myotomy and partial fundoplication. Mean operative time was 180.5 minutes (range 145-264 minutes). Mean length of stay was 1.98 days (range 1-18 days). There were five (10%) perioperative complications but no esophageal leaks. There was a significant difference (P < 0.05) between the preoperative and postoperative dysphagia, chest pain, and regurgitation symptom scores. All patients stated that they were improved postoperatively. Eighty-six per cent rated their outcome as excellent, 10 per cent as good, and 4 per cent as fair. Laparoscopic anterior esophageal myotomy and Toupet fundoplication effectively alleviates dysphagia, regurgitation, and chest pain accompanying achalasia and is associated with high patient satisfaction, a rapid hospital discharge, and few complications. PMID- 11730223 TI - Antiangiogenic therapy with somatostatin receptor-mediated in situ radiation. AB - Tumor growth and the development of metastases require an angiogenic response. Angiogenic vessels uniquely express somatostatin subtype 2 (sst 2) receptors that can transport somatostatin or its analogs into the cell. We hypothesized that radiolabeled somatostatin analogs could inhibit the angiogenic response by selectively destroying proliferating endothelial cells. We evaluated the antiangiogenic effects of 111In-pentetreotide, an sst 2-preferring somatostatin analog in a human vessel model. Disks of human placental vein were embedded in fibrin gels in culture and observed for angiogenic sprouting for 14 days. Vein disks were treated with 111In-pentetreotide (1.5, 15, and 150 microCi/mL) on the day of implantation. Control groups included disks treated with nutrient medium alone, with 111In-chloride, and with unlabeled pentetreotide. The percentage of wells that initiated an angiogenic response and the overall length and density of neovessel sprouts were assessed on Day 14. 111In-pentetreotide treatment did not completely block initiation of the angiogenic response but significantly decreased the growth of neovessels after initiation. Both the receptor-specific Auger electron-induced and nonspecific gamma radiation-mediated effects contributed to the angiotoxicity. PMID- 11730224 TI - Transcutaneous oxygen measurements predict a beneficial response to hyperbaric oxygen therapy in patients with nonhealing wounds and critical limb ischemia. AB - Hyperbaric oxygen (HBO) therapy may be a useful adjunct in the treatment of patients with wounds associated with critical limb ischemia. These patients either cannot undergo a successful bypass or may not heal after vascular reconstruction alone. Identification of patients likely to benefit from HBO is essential before treatment, as this therapy is time-consuming, costly, and not without risk. Transcutaneous oxygen measurements (TCOM) can be used to evaluate the degree of hypoxia in ischemic tissue. In this study we evaluated whether TCOM could be used to identify those patients who would or would not benefit from HBO therapy. Our hypothesis is that a difference in transcutaneous oxygen tension readings measured near the ischemic lesion with the patient breathing room air and while breathing 100 per cent oxygen at ambient pressure may be predictive of wound healing with adjunctive hyperbaric oxygen therapy. Thirty-six patients with critical limb ischemia and nonhealing ulcers were referred for HBO therapy. They were deemed either nonreconstructible from a vascular surgical viewpoint, had failed prior revascularization attempts, or could not achieve complete wound healing even after a successful revascularization. Pretreatment assessment included a room air and post-100 per cent-O2 challenge TCOM reading obtained in the vicinity of the open wound. Hyperbaric oxygen treatments at 2.0 to 2.5 atm were then administered until healing occurred or failure was confirmed. All patients undergoing HBO had a baseline TCOM of <40 torr. Twenty-seven patients had an increase in TCOM of >10 torr with oxygen inhalation at initial evaluation. Of these patients, 19 (70%) healed their wounds with HBO therapy. Conversely the increase in TCOM was <10 torr in nine patients, and only one of these patients (11%) ultimately healed (P < 0.01). Patients with nonhealing ischemic extremity wounds may heal with adjunctive HBO therapy. We can predictably identify patients who are likely to benefit from this modality using TCOM at the time of initial evaluation. An increase of tissue O2 tension of > or =10 torr when breathing pure O2 suggests that the patient may benefit from HBO therapy. Those patients with an increase of <10 torr are unlikely to receive benefit from this treatment modality. PMID- 11730225 TI - Resection of the inferior vena cava for hepatic malignancy. AB - Involvement of the inferior vena cava (IVC) by hepatic tumors, although uncommon, is considered to be unresectable by standard surgical techniques. Recent advances in hepatic surgery have made combined hepatic and vena caval resection possible. The purpose of this study is to describe the surgical techniques and early results of combined resection of the liver and IVC. From 1997 to 2000, 11 patients underwent resection of the IVC along with four to seven liver segments. Resections were carried out for hepatocellular carcinoma (four); colorectal metastases (four); and hepatoblastoma, gastrointestinal stromal tumor metastases, and squamous cell carcinoma in one patient each. Ex vivo procedures were performed twice, and total vascular isolation was used in the nine other cases. The IVC was reconstructed with ringed Gore-Tex tube graft (five), primarily (five), or with Gore-Tex patches (one). There were two early deaths: one from liver failure at 3 weeks and one from sepsis secondary to a perforated segment of small bowel 4 months postresection. One patient with a gastrointestinal stromal tumor died at 32 months of recurrent tumor and one patient with hepatocellular carcinoma is alive with recurrent tumor at 16 months. The remaining patients are alive and disease free with follow-up ranging from 3 to 40 months without evidence of IVC occlusion. Combined resection of the liver and IVC is a formidable undertaking with substantial surgical risk. However, this aggressive surgical approach offers a chance for cure in patients with tumors involving the IVC that would otherwise have a dismal prognosis. PMID- 11730226 TI - Group A Streptococcus (GAS) soft-tissue infections: a lethal organism on the rise. AB - Several reports over the past decade have suggested that there has been an increase in the number of invasive streptococcal infections with young children and the elderly being at the highest risk. We evaluated the incidence of group A Streptococcus (GAS) and compared it with historic data collected at our institution. Prospective data were collected on patients diagnosed with GAS (with and without shock) admitted to a tertiary-care center from July 1995 to July 2000. Each patient was followed by an infectious disease specialist throughout the hospital stay. Definitions of streptococcal toxic shock syndrome (STSS) developed by the Centers for Disease Control and Prevention were used. Thirty eight patients (mean age of 39+/-12) presenting with GAS soft-tissue infections were admitted to our institution over a 5-year period (7.6 patients per year). Fourteen (37%) were diagnosed with STSS. This represents a greater than fourfold increase in the average number of cases per year of patients diagnosed with GAS and a nearly 4.5 times greater increase in the annual number of patients diagnosed with STSS. The overall mortality of patients diagnosed with GAS was 13 per cent, which increased to 36 per cent in patients diagnosed with STSS. We conclude that there has been a significant increase in the incidence of GAS soft tissue infections over the past 5 years at our institution. This may represent a new virulent strain, as the majority of these infections did not occur in typical high-risk patients at the extremes of their lives. Further epidemiologic population-based studies are needed to further delineate the severe nature of this problem. PMID- 11730227 TI - When a hernia is not a hernia: the evaluation of inguinal hernias in the cirrhotic patient. AB - Herniorrhapy in patients with advanced portal hypertension and ascites should be approached with caution, and treated conservatively whenever possible. Cirrhosis increases the risk of significant perioperative complications such as infection, recurrence, and ascites leak. This paper reports two patients operated on for suspected inguinal hernias. The first patient was referred for elective repair of a presumed inguinal hernia before liver transplantation. The second patient presented with a history of an incarcerated inguinal hernia that was previously reduced in the emergency center. After examination by residents and senior faculty the patients were scheduled for elective herniorrhaphy. Intraoperatively no inguinal hernia could be identified in either patient. However, massively dilated veins (1.5-2.0 cm in diameter) were noted entering with the spermatic cord at the internal inguinal ring. In both cases the veins were clamped, transected, and suture ligated at the internal ring. Given the unusual presentation of these dilated veins in both patients we advocate the use of preoperative Doppler ultrasound in patients with cirrhosis and suspected inguinal hernias. PMID- 11730228 TI - Clinical and objective data on spinal cord stimulation for the treatment of severe Raynaud's phenomenon. AB - Ischemic vascular disease of the upper extremity represents a difficult therapeutic problem wherein medical treatment often fails. Epidural spinal cord stimulation has been shown to be an effective alternative in severe peripheral arterial disease. Although this method has been used for nearly two decades only limited experience exists in Raynaud's phenomenon of the upper limbs. In addition objective parameters to prove therapeutic success are not well defined. Herein we describe a patient with severe primary Raynaud's phenomenon over several years who had significant pain relief and complete healing of ischemic digital ulcerations after spinal cord stimulation. Pain level was evaluated using a visual rating scale before and after surgery. Microcirculatory parameters were assessed before and after spinal cord stimulation by capillary microscopy and laser Doppler anemometry. Significant improvement of red blood cell velocity, capillary density, and capillary permeability was demonstrated. At follow-up 18 months after surgery the patient had no complaints and all ulcerations of her fingertips had healed. Spinal cord stimulation appears to be an effective treatment in severe cases of Raynaud's phenomenon and we recommend its use in the case of failed medical therapy. Pain rating and capillary microscopy enable one to assess and visualize the effects of spinal cord stimulation. PMID- 11730229 TI - Traumatic head injury in the anticoagulated elderly patient: a lethal combination. AB - Warfarin is the most common oral anticoagulant used for chronic anticoagulation therapy. Even without any antecedent trauma overanticoagulation can result in intracranial hemorrhage. The triad of anticoagulation with warfarin, age greater than 65 years, and traumatic head injury frequently produces a lethal brain hemorrhage. A retrospective review of more than 2000 patients admitted to the Trauma Service between September 1998 and May 2000 produced 278 patients with head injury and CT-documented intracranial hemorrhage. Of these patients 21 were admitted with an elevated prothrombin time (PT) due to anticoagulation with warfarin. Eighteen patients (86%) were above the age of 70. The most common indications for anticoagulation were atrial fibrillation (71%), deep venous thrombosis (19%), aortic valve replacement (9%), and ischemic cerebral infarcts (9%). Fourteen injuries were the result of a fall, one resulted from a gunshot wound, and one resulted from an assault. The remaining five patients were excluded as their history, workup, and evaluation by neurosurgery suggested a spontaneous bleed leading to fall rather than a fall causing a traumatic bleed. The average Glasgow Coma Score on admission was 11. The average PT and International Normalized Ratio (INR) on admission were 19.2 and 2.99 respectively. Eight of the 16 patients analyzed died. The risk of intracranial hemorrhage with relatively minor head injury is increased dramatically in the anticoagulated patient. A mortality rate of 50 per cent far exceeds the mortality rate in patients with similar head injuries who are not anticoagulated. In addition the risk/benefit equation of anticoagulation for the elderly is more complex and differs from that for younger patients. Perhaps more frequent and judicious monitoring of prothrombin time levels with lower therapeutic ranges (INR 1.5-2) is necessary. PMID- 11730230 TI - An analysis of sestamibi-positive versus -negative patients with primary hyperparathyroidism. AB - We performed this study to evaluate two patient groups with primary hyperparathyroidism depending on whether their abnormal gland(s) could be preoperatively imaged with sestamibi. Patients with primary hyperparathyroidism evaluated by preoperative sestamibi examination from January 1999 to June 2000 were divided into two groups depending on the ability of sestamibi to localize their disease. Records were reviewed to determine pre- and postoperative biochemical data, weight of the excised glands, and total operating room time. When the sestamibi imaging was positive a minimally invasive parathyroidectomy was performed; however, when sestamibi scanning was negative patients underwent a formal bilateral neck exploration. All 40 patients in the sestamibi-positive group and 17 of 18 patients in the sestamibi-negative group were cured of their primary hyperparathyroidism as a result of surgery. Sestamibi scanning with a minimally invasive parathyroidectomy shortens operating room time and is most effective when adenomas are large. The results of this study suggest that strategies to preoperatively increase the activity of adenomas may improve the sensitivity of sestamibi scan localization of parathyroid adenomas. PMID- 11730231 TI - Videoscopic heller myotomy as first-line therapy for severe achalasia. AB - To many nonsurgeons myotomy is considered an excessively invasive treatment for achalasia and has become a salvage procedure when esophageal dilation and botulinum toxin (botox) injections fail. We sought to examine our experience with videoscopic Heller myotomy to determine whether preoperative therapy predicts perioperative complications and long-term outcome. Videoscopic Heller myotomy was undertaken in 111 patients with achalasia between June 1992 and May 2000. Intraoperative endoscopy was used in all patients. Fundoplication was used selectively for patients with large hiatal hernias or as part of repair of esophageal perforation. Patients were asked to grade their dysphagia and reflux symptoms before and after myotomy on a scale of 0 (no symptoms) to 5 (severe symptoms). Patients were also asked to rate their outcome as excellent (no symptoms), good (greatly improved), fair (slightly improved), or poor (not improved) compared with their preoperative status. Patients were stratified on the basis of preoperative intervention (botox, pneumatic dilation, botox and pneumatic dilation, or no botox or dilation) and compared. Previous pneumatic dilation and/or botox injection had been undertaken before operation in 88 (79%) patients whereas 23 (21%) patients had no invasive preoperative therapy. The overall mean preoperative dysphagia score was 4.8+/-0.8 and mean preoperative reflux score was 3.3+/-2.1. Groups of patients undergoing preoperative interventions were similar to those patients not undergoing preoperative interventions in terms of preoperative symptoms, dysphagia scores, and reflux scores. Postoperative complications (13%) and perforations (8%) were slightly more common in patients who had undergone preoperative botox or dilation (P = not significant). Subjectively, operative myotomy was more difficult in patients who had preoperative botox or dilation. Patients had significant improvement in dysphagia, dysphagia score, reflux score, emesis/ regurgitation, and chest pain (P < 0.05) regardless of preoperative intervention. After myotomy patients who had never undergone botox or pneumatic dilation were less likely to have mild dysphagia compared with those with previous botox injections (30% vs 53%; P = 0.09), previous dilations (30% vs 54%; P = 0.09), or both (30% vs 59%; P = 0.04). As well, dysphagia scores were better if no preoperative therapy had been undertaken: botox 0.8+/-1.3, dilation 1.0+/-1.4, botox and dilation 1.0+/-1.3, and no therapy 0.3+/-0.7 (P < 0.05). Overall 97 per cent of patients stated that their symptoms were improved although more patients tended to have excellent or good outcomes if no preoperative intervention was undertaken (91%) compared with patients undergoing preoperative botox (86%), dilation (83%), or both (82%) (P = not significant). We conclude that videoscopic Heller myotomy is safe and efficacious particularly in patients who have not undergone previous endoscopic interventions. The difference in patients' outcomes based on preoperative therapy may be related to a less difficult operation in patients who forgo endoscopic therapy and elect to undergo early myotomy. Although videoscopic Heller myotomy provides good outcomes as a salvage procedure after failed dilations and/or botox injections for achalasia we advocate it as first-line therapy in reasonable operative candidates. PMID- 11730232 TI - Does alcohol intoxication alter the assessment and outcome of "observation status" trauma patients? AB - We assessed the effect of blood alcohol concentration (BAC) on the evaluation, outcome, and hospital charges of our observation-status trauma patient population. We conducted a retrospective study over 18 months; any patient initially admitted with <24-hour observation status, Glasgow Coma Score of 15, and negative drug screen was eligible. Patients were divided on the basis of BAC (BAC+ = >80 mg/dL; BAC- = <80 mg/dL). Two hundred twenty-six patients were observed during the study (2765 admissions). For the 66 BAC+ patients (range 90 392 mg/dL) there was a strong male predominance. There was no difference in diagnostic evaluation schema, delayed diagnosis, complications, cost, or conversions to full admission between the groups. We conclude that evaluation, outcome, and charges of observation trauma patients are the same regardless of BAC. Intoxication did not mask injury; therefore BAC+ patients do not require observation on the sole basis of intoxication if their evaluation is otherwise negative. PMID- 11730233 TI - Tensile strength of wound closure with cyanoacrylate glue. AB - 2-Octyl cyanoacrylate tissue adhesive is increasingly being used for closure of traumatic lacerations. Data regarding the strength of incisions closed with 2 octyl cyanoacrylate are limited. We compared the strength of disruption of closure with glue with that of more conventional methods of wound closure. Segments of fresh porcine skin measuring 3.5 x 10 cm were approximated by one of four methods: 1) 2-octyl cyanoacrylate glue, 2) surgical staples, 3) 0.5 inch Steri-Strips, and 4) interrupted 4-0 poliglecaprone 25 sutures in a subcuticular fashion. Fifteen specimens were used to test each type of closure. The strength of closure was tested on an Instron 4502 tensionometer. The peak force required for disruption of the closure was recorded and the strength of the closure was compared. Staples provided the strongest closure. Skin glue proved superior to Steri-Strips but inferior to stapled closure. The difference between skin glue and suture closure was not statistically significant (P = 0.12). Patterns of failure differed between the groups. Skin glue failed because of disruption of the skin-glue interface. 2-Octyl cyanoacrylate glue provides a wound closure that is similar to closure with an interrupted subcuticular absorbable suture. This study validates the clinical use of skin glue for closure of surgical incisions. The technique should be used with caution in areas of the body that are subject to tension. PMID- 11730234 TI - Physical activity in aging: changes in patterns and their relationship to health and function. AB - Sedentary behavior is an important risk factor for chronic disease morbidity and mortality in aging. However, there is a limited amount of information on the type and amount of activity needed to promote optimal health and function in older people. The purpose of this review is to describe the change in patterns of habitual physical activity in aging and the relationship of these changes to physical function and selected chronic diseases. We undertook a literature review of large population-based studies of physical activity in older people, and there is encouraging evidence that moderate levels of physical activity may provide protection from certain chronic diseases. Additionally, substantial health effects can be accrued independent of the fitness effects achieved through sustained vigorous activity. Thus, regular participation (i.e., 30 minutes/day on most days of the week) in activities of moderate intensity (such as walking, climbing stairs, biking, or yardwork/gardening), which increase accumulated daily energy expenditure and maintain muscular strength, but may not be of sufficient intensity for improving fitness, should be encouraged in older adults. Public policy should focus on ways of increasing volitional and lifestyle activity in older people, as well as on increasing the availability and accessibility of senior and community center programs for promoting physical activity throughout the life span. PMID- 11730235 TI - Physical activity and quality of life in older adults. AB - Although there has been increased research and clinical attention given to the effects that physical activity has on quality of life among older adults, there is a lack of consistency surrounding the use of this term. As a result, attempts to examine what causes change in quality of life have been limited. This article critically reviews the literature on physical activity and quality of life in older adults. In so doing, attention is given to both quality of life as a psychological construct represented by life satisfaction as well as a clinical and geriatric outcome represented by the core dimensions of health status or health-related quality of life. The literature is also examined to identify potential mediators and moderators in the physical activity and quality-of-life relationship. Discussion of possible mediating variables reinforces the important role of perception when considering the beneficial effects that physical activity has on quality of life. From a public health perspective, understanding what may cause change in quality of life has significant implications for the design, implementation, and promotion of physical activity programs for older adults. PMID- 11730236 TI - Interventions to promote physical activity by older adults. AB - Physical inactivity has been established to be an independent risk factor for a range of chronic diseases and conditions that threaten the health of the nation. However, only a minority of the population is currently meeting the recommended levels of regular physical activity, which have been linked with important health and quality-of-life benefits. Older adults are at particular risk for leading sedentary lifestyles. This article provides an overview of factors associated with physical activity for older adults and also describes potentially promising interventions for promoting regular physical activity in this growing population segment. Examples of interventions undertaken at personal and interpersonal as well as broader levels of analysis (e.g., environmental) are provided. Major issues currently facing the field are discussed, including the ongoing challenge of developing assessment tools that are sensitive to the more moderate-intensity physical activities favored by older adults and the formidable task of combining clinical approaches with environmental and policy strategies aimed at combating this public health problem. PMID- 11730237 TI - Nutrition and health promotion in older adults. AB - During recent decades, the concept of health promotion has become a legitimate part of health care because of the aging of the postwar baby boom generation. As this population ages, the potential strain on health care systems will increase because the greatest use of health care services occurs during the last years of life. In older adults there are many correctable health factors that can be assessed through screening protocols. Hypertension, cholesterol, hearing, vision, diabetes, and cancer screening are well integrated into health promotion programs; nutrition promotion programs are not as well integrated. Reluctance to develop health promotion programs for older adults exists because of a perception that they would not follow such plans or change their lifestyles. However, longitudinal studies have shown that health promotion activities extend the number of years of health in older people although the relationship weakens in older age. Changes in diet and exercise patterns are most effective in the prevention of nutrition-related conditions when they are instituted early in life, but positive effects can occur at any age. If nutritional interventions are instituted early, a substantial reduction in health care expenditures may result from a decrease in the incidence or the delayed onset of these conditions. Changes in behaviors (reducing salt and fat intake) were positively associated with a belief that consuming a healthful diet would contribute to better health. The use of a variety of adult education theories and models will enhance behavior changes that lead to more healthful habits and enable a health educator to be successful in effecting change. PMID- 11730238 TI - Nutrition and quality of life in older adults. AB - Good nutrition promotes health-related quality of life (HRQOL) by averting malnutrition, preventing dietary deficiency disease and promoting optimal functioning. However, definitions of quality of life also encompass life satisfaction and both physical and mental well-being. Nutrition and diet have not been a part of mainstream research on quality of life and are not included among key quality of life domains. This article explores connections between diet and nutritional status in relation to HRQOL measures and overall well-being among older adults. PMID- 11730239 TI - Dietary intake, dietary patterns, and changes with age: an epidemiological perspective. AB - Cohort and cross-sectional data were reviewed to describe the changes in dietary intake with age. Total energy intake decreases varied substantially with age, by 1000 to 1200 kcal in men and by 600 to 800 kcal in women. This resulted in concomitant declines in most nutrient intakes. For some nutrients, substantial numbers of older Americans consumed only one fifth to one third of the recommended dietary allowance. For most nutrients, research is lacking with which to judge the health impact of reduced nutrient consumption with age, although there is some evidence of an age-related decline in absorptive and metabolic function. With the aging of the population, more research is needed on nutrient requirements and health outcomes, and public health efforts are needed to increase physical activity and food intake among older people. PMID- 11730240 TI - Physical activity and parameters of aging: a physiological perspective. AB - Increasing age is associated with a decline in fat-free mass. The question is whether age-related changes in body composition can be delayed by an active life style. This analysis includes data where physical activity was assessed with doubly labeled water and body composition with hydrodensitometry or isotope dilution. Subjects were 136 women and 180 men over 20 years, who were tested in Maastricht University between 1983 and 1998. Increasing age was associated with lower activity levels and lower fat-free mass. After controlling for age there was no longer any association between physical activity and fat-free mass. A few exercise intervention studies showed that elderly subjects compensate for exercise training by a decline in spontaneous physical activity, in contrast to younger subjects. Although no effect of habitual activity level on changes in body composition are observed, training has a positive effect on muscle function. Elderly subjects with relatively high levels of physical activity are not different from those with low activity levels, as far as fat-free mass and fat mass are concerned. However, training might delay the age-induced impairment of personal mobility associated with a reduction in physical activity. PMID- 11730241 TI - Decreased food intake with aging. AB - There is a physiological decline in food intake with aging. The reasons for the decline in food intake are multifactorial and involve both peripheral and central mechanisms. Altered hedonic qualities of food occur due to alterations in taste and, more particularly, smell with aging. A decline in adaptive relaxation of the fundus of the stomach and an increased rate of antral filling appear to play a role in the early satiation seen in many older persons. Cholecystokinin levels are increased with aging and older persons are more sensitive to the satiating effects of this gut hormone. The decline in testosterone levels in older males leads to increased leptin levels and this may explain the greater decline in food intake with aging in the male. Within the hypothalamus, decreased activity of both the dynorphin (kappa opioid) and neuropeptide Y systems occurs in aging rodents. Cytokines are potent anorectic agents. Many older persons have mild inflammatory disorders that lead to anorexia. Exercise may increase food intake in older persons. PMID- 11730242 TI - Nutrition, physical activity, and quality of life in older adults: summary. AB - If health-related quality of life--and not longevity--is the key goal for health promotion, then it is captured only partly by the existing mortality and morbidity indexes. Researchers now urge that government agencies and health care providers begin collecting quality-of-life data on the populations they serve. Adding life to years, not years to life, is the current agenda for productive and successful aging. Policies and programs on aging are increasingly focused on identifying ways to improve quality of life and health status rather than just extending life span. In the Healthy People 2000 report, the chief goal of health promotion was to increase the span of healthy life. The focus was on mortality and morbidity data and symptom checklists as the principal measures of ill health. In contrast, the new emphasis in the Healthy People 2010 report is on quality of life and overall well-being. Helping people to increase life expectancy and improve their quality of life is the primary goal of the Healthy People 2010 report. The authors of this special issue of the Journals of Gerontology: Biological Sciences and Medical Sciences are united in the belief that optimal nutrition and physical activity make a significant contribution to the overall quality of life at any age and especially for older adults. The key research challenge lies in deciding which aspects of improved fitness, nutrition, and diet contribute the most to quality-of-life measures. We have attempted to provide a comprehensive review of research on exercise, nutrition, diet, and health in elderly adults. Past studies on diet, nutrition, and fitness have largely addressed biomedical outcomes, pointing to substantial benefits in physical functioning, remission of disease symptoms, and improved health. This special issue goes a step further in assessing the effect of improved nutrition and physical activity on the global quality of life and its four principal domains. Although links between diet and exercise and chronic disease risks have been well documented, more needs to be known about motivations for behavioral change and perceived benefits as assessed using quality-of-life measures. No single segment of our society can benefit more from regularly performed exercise and improved diet than elderly adults. These important articles provide a link between diet and exercise and quality-of-life issues, as outlined in the Healthy People 2010 report. PMID- 11730243 TI - The First International Conference on Osteoporosis in Men. Siena, Italy, February 23-25, 2001. PMID- 11730245 TI - Mortality, morbidity, and assessment of fracture risk in male osteoporosis. AB - Several studies show that for most clinical fractures the mortality is higher in men than in women, also compared with the general population. However, there are still uncertainties whether this is true for the vertebral deformities. There are few studies comparing men and women but there does not seem to be any difference in morbidity between men and women even though data here are scarce. Several of the risk factors that have been identified in women are also important in men. However, there are problems comparing men and women, such as recreational activity, where men have more strenuous activities. The same for alcohol intake and for BMD. Larger studies in men are needed. At present there are no diagnosis or intervention thresholds for men. Indication thresholds should be determined for those who have a high risk of fractures and should be based on a combination of risk factors in which bone mineral density (BMD) is the most important. This review of papers has dealt with women and men within the same study to compare differences or similarities. PMID- 11730244 TI - The prevalence of osteoporosis: gender and racial comparison. AB - Osteoporosis is common among the growing population of older men: almost 20% of men > or = 50 years old have osteoporosis of the hip, spine, or wrist. However, the exact estimate depends on the approach taken to normalize for bone size, the specific skeletal site assessed, and the diagnostic criteria used. Bone mineral density (BMD, g/cm2) by DXA is 12-25% greater in men than women, but bone mineral apparent density (g/cm3) is similar in the two sexes. This correction for skeletal size largely eliminates apparent differences in areal BMD between the races and also reduces the apparent effects on BMD of age. The particular skeletal site that is assessed has an important influence on the prevalence of osteoporosis (sex-specific BMD T-score less than -2.5) in men which varies from 0 to 36%, depending on the site, and from 2% to 45% in postmenopausal women. The discrepancies relate mainly to different patterns of bone loss at the various sites, but estimates are also affected by the specific young normal means and standard deviations (SD) used to calculate the T-scores. A greater mean and smaller SD among normal young men in Rochester, MN produced a higher prevalence of osteoporosis at the femoral neck (22% vs 7%) compared with estimates for white men from the Third National Health and Nutritional Examination Survey; use of female normal values further reduced osteoporosis prevalence at the hip in white, Hispanic, and African-American men to 4%, 2%, and 3%, respectively, compared with 20% for white women in the United States. By contrast, fracture risk is similar for men and women at any given level of BMD. These observations reinforce current efforts to move away from osteoporosis prevalence and toward absolute fracture risk as the main basis for clinical treatment decisions. PMID- 11730246 TI - Androgens: basic biology and clinical implication. AB - Although androgens have been considered essential modulators of bone biology in men, recent studies have indicated that estrogen may have an important, if not dominant, role. Nevertheless, there is strong evidence that androgens have independent skeletal actions. Nonaromatizable androgens influence a variety of aspects of bone cell biology and are capable of modulating bone remodeling and bone mass. It appears that androgens are particularly important in the control of periosteal bone formation, an effect that might underlie the gender difference in bone size. Alterations in androgen receptor function affect bone metabolism, and new information suggests that androgens modulate receptor homeostasis. The clinical implications of androgen effects, and how they interact with those of estrogens, are somewhat unclear. It is likely that overall bone homeostasis and gender differences depend on a combination of androgenic and estrogenic actions. Androgens may well provide advantages in the prevention and therapy of metabolic bone disorders in both men and women. PMID- 11730247 TI - Estrogens and bone health in men. AB - It has generally been held that estrogen and testosterone are the major sex steroids regulating bone metabolism in women and men, respectively. However, the description of several "experiments of nature" led to a reconsideration of this notion. Thus, a male carrying homozygous mutations in the ER-alpha gene and two males with homozygous mutations in the aromatase gene had osteopenia, unfused epiphyses, and elevated indices of bone turnover. Though these findings indicated that estrogen plays a role in regulating the male skeleton, they left unresolved the issue of whether estrogen acted on the male skeleton mainly to enhance bone mass acquisition during growth and maturation, or whether it also acted to retard bone loss in aging individuals. To address this issue, several cross-sectional observational studies have related bone mineral density (BMD) to sex steroids in elderly men, and found that estrogen correlated better than testosterone with BMD. In addition, recent longitudinal studies from our group indicate that bioavailable estrogen correlated better than testosterone both with the gain in BMD in young men and with loss of BMD in elderly men. These observational studies do not, however, prove causality, which requires direct interventional studies. Thus, we eliminated endogenous testosterone and estrogen production in 59 elderly men (mean age 68 years), studied them first under conditions of physiologic testosterone and estrogen replacement, and then assessed the impact on bone turnover of withdrawing both testosterone and estrogen, withdrawing only testosterone, only estrogen, or continuing both. We found that estrogen played the major role in regulating bone resorption in these men, and that both estrogen and testosterone were important in maintaining bone formation. Collectively then, these findings indicate that estrogen plays a dominant role in regulating the male skeleton. PMID- 11730248 TI - Secondary causes of osteoporosis in men. AB - Important underlying causes of osteoporotic fracture in men include glucocorticoid therapy, low body weight, and reduced physical activity. Tobacco and alcohol use have been consistently identified as risk factors for vertebral fracture but there is less evidence that they contribute to hip fracture. Clinically overt hypogonadism is a strong risk factor for osteoporosis in men; however, the role of more subtle subclinical changes, as defined by biochemical criteria, remains to be established. The high comorbidity associated with osteoporosis, particularly in elderly men, contributes to fracture risk both through effects on bone mass and risk of falling. The management of osteoporosis in men includes diagnosis of and, where possible, correction of underlying contributory causes. Evidence from recent randomized controlled trials indicates that bisphosphonates are effective in the prevention of glucocorticoid-induced osteoporosis in men but the optimal criteria for selection of individuals for treatment requires further study. PMID- 11730249 TI - Skeletal effects of exercise in men. AB - No prospective randomized study with fracture as end point exists in men. Data from retrospective and prospective observational and case control studies suggest that activity in men is associated with reduced fracture risk. This may be correct, but consistently replicated sampling bias may produce the same observation, as these studies are hypothesis generating, never hypothesis testing. Higher musculoskeletal mass, better health, coordination, and less tendency to fall may lead to exercise, not the reverse. It would be extremely difficult to conduct an exercise intervention study with fracture as the end point because of the large cohorts needed. However, showing a positive effect on surrogate end points for fractures as increased bone mass or reduced fall frequency would support the notion that exercise has a fracture-protective effect. Exercise during growth seems to build a larger and stronger skeleton in boys. However, cessation of exercise is its Achilles heel; biologically important increased peak bone mass or improvement in muscle strength achieved by exercise during growth may be eroded in retirement, leaving no biological significant benefits in old age when fractures occur. Exercise during adulthood may prevent bone loss or produce a small increase in BMD of a few percent, probably a non biologically significant increase in reducing the fracture risk in elderly men. However, exercise in adults seems to increase muscle strength and improve balance, with reduced fall frequency as the result and maybe reduced frequency of injuries also. The effects of exercise on bone size, shape, and architecture during growth and adolescence must be defined as well as the effect of muscle function, fall frequency, and frequency of injurious falls in elderly men. Also, continued low level of exercise may maintain some of the benefit after more vigorous activity level during younger years, but dose-response relationships need to quantified. Additionally, the null hypothesis that exercise has no effect on fracture rates in old age cannot be rejected by any published data. The proof rests on demonstration of a reduction in spine and hip fractures in well-designed and executed prospective, open-randomized studies, none of which exist. Our inability to answer these questions should be acknowledged before recommendations are made at the community level. PMID- 11730250 TI - Genetics of male osteoporosis. AB - In the past years, several epidemiological and clinical observations have underlined the importance of genetics in the pathogenesis of both female and male osteoporosis. It has been estimated that from 50 to 80% of the inter-individual variability in bone mass is genetically determined. In rare instances, osteoporosis in men could be inherited in a simple Mendelian pattern. Examples of this include familial osteoporotic syndromes due to mutations in the aromatase and ER alpha genes. Families have also been described in which high bone mass is inherited as an autosomal dominant trait, consistent with the effect of a single gene located on chromosome 11. However, except for these rare conditions, osteoporosis has to be considered a multifactorial disease in which genetic determinants are modulated by hormonal, environmental, and nutritional factors. The genetic effect on bone may also be gender- and site-specific, with different genes regulating bone density at different skeletal sites in males and females. To date, most of the work on the genetics of osteoporosis has focused on women. In some studies, polymorphisms at the IGF-I, VDR, COLI-alpha1, ER alpha, and aromatase gene have been recently shown to predict BMD variation and osteoporotic risk in males. These observations remain to be confirmed by other independent studies. Other candidate genes, are still awaiting mapping and identification. PMID- 11730251 TI - During aging, men lose less bone than women because they gain more periosteal bone, not because they resorb less endosteal bone. AB - Bone size is greater in young men than in women but the load imposed is also greater. Therefore, the load per unit area (stress) on the vertebral body is the same in men and women. Fractures are uncommon in young adulthood because volumetric bone mineral density (vBMD) is normal so that the stress imposed on the bone is well below its strength in both sexes. The gender difference in bone fragility emerges during aging because the biomechanical relationship between the stress on bone and its strength changes in men and women. Periosteal apposition occurs in both genders during aging but more in men than in women so that vertebral cross-sectional area (CSA) increases more in men than women. Consequently, the stress on bone decreases more in men than women. In addition, periosteal apposition offsets endosteal bone loss more in men than in women so that net bone loss is less in men than women. Thus, the stress on the bone decreases more and the strength of the bone decreases less in men than in women. The ratio of the stress on bone to its strength can be quantitated and is below unity in young men and women. During aging this ratio increases to approach or exceed unity, the value at which fracture risk is high because the stress on bone is similar to its strength. As the stress decreases more and strength decreases less in men, the ratio increases less in men than in women so that in old age only 2-3% of men but 20% of women have this stress to strength ratio exceeding unity. Men and women with spine fractures have the same vBMD and same stress to strength ratio which is no different from unity. Fewer men than women are at risk for fracture because fewer men than women have these structural determinants of bone strength below a level at which the loads exceed the bone's ability to tolerate them. PMID- 11730252 TI - Histomorphometric approach of bone loss in men. AB - From a review of the published literature the authors have described the histomorphometric changes observed in human iliac bone corresponding to the age related bone loss in 'males and in "idiopathic" male osteoporosis. Age-related bone loss of about 30% is histomorphometrically characterized in males by a reduction in iliac cancellous bone volume between ages 20 and 70-80 years, less marked than in women (-42%). The age-related thinning of trabecular packets wall width is the same between males and females, but trabecular separation is lower in aged males than in aged females, suggesting a better preservation of the trabecular microarchitecture in elderly males. Trabecular eroded surfaces increase with age in females but not in males. The rare studies comparing bone formation parameters in males and females did not show any major difference, with a trend for a decrease in mineralization apposition rate and an increase in activation frequency in both sexes. Bone histomorphometry in "idiopathic" male osteoporosis is characterized by a mean decrease in cancellous bone volume of 35% compared with age-matched controls, similar to the mean decrease observed in females (-38%). Most, but not all authors have found an increase in bone resorption parameters and a decrease in bone static and dynamic formation indices (osteoid surfaces, mineralizing surfaces, bone formation rate). PMID- 11730253 TI - Bone microarchitecture and bone fragility in men: DXA and histomorphometry in humans and in the orchidectomized rat model. AB - In men, the risk of fragility fractures increases as bone mineral declines but there is an overlap in the bone mineral density (BMD) measurements between patients with and those without fractures. Biomechanical competence of trabecular (Tb) bone depends on the amount of bone and on microarchitecture. We have developed new histomorphometric methods for evaluating microarchitecture on histological sections. These methods were used in the orchidectomized male rat (ORX--a model of hypogonadism-induced osteoporosis) and on transiliac bone biopsies performed in male osteoporotic patients. ORX rats were studied at 2, 4, 8, and 16 weeks post-ORX. Bone mineral content (BMC) was reduced at 16 weeks. Trabecular bone volume (BV/TV) was significantly decreased from the 4th week. Differences in the sensitivity of the methods were found. Fractal dimension was modified as early as 2 weeks and appeared the most potent descriptor of Tb disorganization. The architectural changes in this model mimic those observed in hypogonadic men. We examined the relationships among BMD, micro-architecture, and vertebral fracture in 108 men with lumbar osteopenia (T-score <-2.5). At least one vertebral fracture was observed in 62 patients and none in 46 patients. After adjusting for age, body mass index (BMI), and BMD, there was no significant difference between the two groups in BV/TV, Tb.Th, and Star volume. In contrast, the mean values of ICI and Tb.Sp were significantly higher whereas Tb.N and nodes were lower in patients with vertebral fracture. Logistic regression analysis showed that ICI, strut analysis, and Tb.N were significant predictors of the presence of vertebral fracture: odds ratios for an alteration of I SD ranged from 1.7 for nodes to 3.2 for ICI. These results strongly suggest that bone Tb microarchitecture is a major and independent determinant of vertebral fracture in men with osteoporosis. PMID- 11730254 TI - Diagnosis of osteoporosis and fracture threshold in men. AB - This review examines the hypothesis that there are gender differences in the relationship between bone mineral density (BMD) and fracture risk. Prospective studies of spine and hip fracture risk drawn from population samples suggest that the fracture risk increases as BMD decreases in men in the same way as that described in women. Moreover, for any given BMD at the spine, heel, or proximal femur the risk in men is similar to that in women. Although data are limited, prior fragility fractures increase subsequent fracture risks in both men and women. These studies suggest that the same BMD criteria used to diagnose osteoporosis in women can be applied to men. PMID- 11730255 TI - The assessment of bone mass in men. AB - Bone mineral density (BMD) is widely used in postmenopausal women to identify who should be given therapy for prevention and treatment of osteoporosis and to monitor the efficacy of treatment. There is still uncertainty about how to interpret BMD in men, and few prospective studies exist on the relationship between BMD and fracture risk. Men should be considered for measurement of BMD if they have suffered low trauma fractures, have prevalent vertebral deformities, have radiographic osteopenia, are over age 75, or have conditions that increase their risk for bone loss, such as hypogonadism, glucocorticoid use, or generally poor health. There is insufficient information to recommend a more widespread BMD screening. The World Health Organization has developed criteria for interpreting BMD which are widely used. Patients with BMD at least 2.5 SD below the young adult mean (T-score < -2.5) have osteoporosis, and those with BMD between 1 and 2.5 SD below the young adult mean (-2.5 < T-score < -1.0) have osteopenia. However, the BMD criteria that should be used to identify men in need of therapeutic intervention are still debated. Using male-specific hip BMD cutoffs, approximately 3-6% of U.S. men 50 years and older were estimated to have osteoporosis and 28-47% to have osteopenia. The corresponding figures in women were 13-18% with osteoporosis and 37-50% with osteopenia. Greater accumulation of skeletal mass during growth, slower rate of bone loss, and shorter life expectancy in men contribute to the lower prevalence of osteoporosis relative to women. PMID- 11730256 TI - Use of quantitative ultrasound densitometry (QUS) in male osteoporosis. AB - Bone densitometry is a key factor in the early diagnosis of osteoporotic bone disorders. Cut-off values for WHO classification for male osteoporosis and all densitometry techniques such as dual x-ray absorptiometry (DXA), quantitative ultrasound (QUS), and quantitative computed tomography (QCT) need to be developed. Hereby, QCT, DXA, and QUS are equivalent methods in the prospective assessment of fracture risk. Where men and women have similar BMD values, they also have similar fracture probability. QUS has several advantages compared to the radiological devices. The QUS systems, which are commercially available at present, are non-invasive methods, which are safe, simple, free of radiation, portable, and relatively inexpensive. QUS can be measured at the calcaneus and phalanges or with multi-site systems. Phalangeal ultrasound is especially useful as being easily accessible. PMID- 11730257 TI - Biochemical markers of bone turnover in men. AB - Although osteoporosis in men has been recently recognized as a public health problem, the mechanisms leading to bone loss are still poorly understood. Longitudinal studies of bone mineral density suggest an acceleration of bone loss after 70 years of age. Histomorphometric data concerning age-related changes of bone turnover in men are limited, including few men over 70 years and have been restricted to the trabecular envelope of bone biopsies. Most measurements of biochemical markers of bone turnover have been performed in small cohorts of limited age range, and results obtained in large cohorts are scanty. Levels of markers of bone formation and of bone resorption are very high in men aged 20-30 years which corresponds to the late phase of formation of peak bone mass, and then declines, reaching their lowest levels between 50 and 60 years. Data on bone turnover markers in elderly men are discordant. Concentrations of bone formation markers remain stable, decrease slightly, or even increase marginally. Markers of bone resorption increase in some studies, mainly after 70 years of age, in line with acceleration of bone loss in this age range. This discordance between studies can result from different reasons. The increase of bone turnover may be limited to a subgroup of elderly men. In addition, urinary levels of bone resorption markers depend on the rate of bone turnover, on pre-renal and renal catabolism of peptides released from bone matrix, on glomerular filtration rate, as well as unit of expression of their results (per 24 hours per urinary creatinine mass, per glomerular filtrate volume). In elderly men, biochemical bone markers are negatively correlated with bone mineral density. Longitudinal studies are not yet available on the relationship among bone turnover markers, rate of bone loss, and fracture. In conclusion, in elderly men, age-related bone loss seems to result from increased bone resorption which is not matched by increased bone formation. Thus, antiresorptive therapy may be of interest in the prevention and treatment of osteoporosis in men. Further studies are necessary to determine if bone resorption markers predict the risk of fragility fractures in elderly men. PMID- 11730258 TI - Androgen replacement in aging men. AB - Serum testosterone and estradiol decline with advancing age in men, but the contribution of these changes to age-related bone loss remains unclear. Most studies show a closer relationship between bone density and serum estradiol than with serum testosterone. Nevertheless, hypogonadism is widely considered to be an important cause of male osteoporosis, occurring in up to 20% of men with vertebral fractures and 50% with hip fractures. Bone loss in hypogonadal men has been attributed to androgen and estrogen deficiency, low 1,25 dihydroxyvitamin D concentrations, malabsorption of calcium, and reduced circulating calcitonin. These abnormalities are corrected by testosterone replacement, which also decreases bone resorption and stimulates bone mineralization. Long-term testosterone replacement increases spine bone density by over 25%, although much of the improvement occurs in the first few years of treatment. There appears to be an inverse relationship between the basal serum testosterone and the increase in bone density observed with testosterone treatment, such that little change is seen in men with normal basal concentration. In contrast, an observational study in eugonadal men with vertebral fractures showed an increase in spine bone density of 5% after 6 months' treatment with testosterone, probably mediated by an increase in serum estradiol and a reduction in bone resorption. Although testosterone has beneficial effects on the skeleton, there may be adverse effects on the prostate and cardiovascular risk factors. Further studies are required to confirm the safety and efficacy of testosterone replacement in hypogonadal and eugonadal men with osteoporosis. PMID- 11730259 TI - Alendronate for the treatment of osteoporosis in men. AB - The incidence of osteoporotic fracture in males is approximately one-third of that observed in women, but information on specific therapies is almost exclusively limited to bisphosphonate alendronate. The most important study with this compound included 241 men, randomized to receive either alendronate 10 mg/day or placebo. In another study 134 men were given either 10 mg alendronate or alfacalcidiol 1 microg/day. After 24 months, the treatment with alendronate bone mineral density (BMD) significantly increased in both studies by 7-10% at the lumbar spine and by 2.5-5.2% at the femoral neck. These changes were associated with decreases in vertebral fracture rate and in stature loss, both statistically significant when the data of the two trials were meta-analysed. The BMD changes after alendronate therapy were comparable to those observed in postmenopausal osteoporosis. This was confirmed in a trial specifically designed to compare alendronate efficacy in men and postmenopausal women with either primary or secondary osteoporosis. Gender-comparative efficacy data can also be inferred from clinical trials in glucocorticoid-induced osteoporosis of alendronate, risedronate, and etidronate, carried out in both women and men. By combining the results of all these trials, bisphosphonate efficacy in terms of both BMD changes and fracture incidence appears to be moderate in premenopausal women but quite obvious and comparable in males and postmenopausal women. PMID- 11730260 TI - Risedronate increases bone density and reduces vertebral fracture risk within one year in men on corticosteroid therapy. AB - Limited information is available on the effect of bisphosphonates in men receiving corticosteroid therapy. We studied 184 men among the patients enrolled in two, double-blind, placebo-controlled, 1-year studies with similar protocols. The studies evaluated the effects of risedronate in patients beginning corticosteroid treatment at a dose of at least 7.5 mg per day of prednisone or equivalent (prevention study) or continuing long-term treatment of corticosteroid at that dose (treatment study). The men received either placebo or risedronate (2.5 mg or 5 mg) daily, along with calcium supplementation (500-1000 mg). Endpoints included differences in bone mineral density (BMD) at the lumbar spine, femoral neck, and femoral trochanter, assessment of vertebral fractures, changes in biochemical markers of bone turnover, and overall safety. In the treatment study, risedronate 5 mg significantly (P < 0.01) increased lumbar spine BMD by 4.8% at the lumbar spine, 2.1% at the femoral neck, and 2.6% at the femoral trochanter compared with baseline values. In the prevention study, bone loss was prevented with risedronate 5 mg; in the placebo group, BMD decreased significantly (P < 0.01) by 3.4%, 3.3%, and 3.4% in the lumbar spine, femoral neck, and trochanter, respectively, at 1 year. The differences between risedronate 5 mg and placebo groups were significant at all skeletal sites in the prevention study (P < 0.01) and at the lumbar spine in the treatment study (P < 0.001). The 2.5 mg dose also had a positive effect on BMD, although of a lesser magnitude than the 5 mg dose. When the data from the two studies were combined, the incidence of vertebral fractures decreased 82.4% (95% confidence interval, 36.6%-95.1%) in the pooled risedronate groups compared with placebo (P = 0.008). Risedronate was well tolerated in men, with a similar incidence of upper gastrointestinal adverse events in the placebo and treatment groups. Daily treatment with risedronate increases bone density and decreases vertebral fracture risk within 1 year in men receiving corticosteroid therapy. PMID- 11730261 TI - Therapy of male osteoporosis with parathyroid hormone. AB - Definable causes of male osteoporosis account for only about 60% of the osteoporotic population. Those for whom no etiology is readily apparent are said to have primary or idiopathic male osteoporosis. In these individuals, histomorphometric studies indicate that this is a disorder that is more typically characterized by low turnover. Although antiresorptive agents such as alendronate have been shown to increase bone mass in men, the rationale for an anabolic agent that can stimulate bone formation is clear. The most attractive anabolic agent at this time is parathyroid hormone (PTH) administered in low dosage and intermittently. Such regimens in experimental animals have been associated with marked gains in bone mass. Slovik et al. showed that parathyroid hormone can increase vertebral bone mass in men with idiopathic osteoporosis. We have conducted the first controlled, randomized, double-blind study of PTH in men with idiopathic osteoporosis. Twenty-three men, 30-68 years old (50 +/- 1.9) with Z scores less than -2.0 were assigned to a placebo (n = 13) or treatment (n = 10) arm. After 18 months, those who received PTH showed a 13.5 +/- 3% increase in bone mass, significantly greater than the placebo group whose bone density did not change. Femoral neck bone density increased significantly by 2.9 +/- 1.5%. The distal radius site did not change. During an open label extension for an additional 12 months, there was no further increase in bone density in the lumbar spine but the femoral neck continued to show gains. Markers of bone formation and resorption increased in the PTH arm reaching a peak between 9 and 12 months of therapy and declining thereafter. Parathyroid hormone was well tolerated. These results suggest that low-dose intermittent PTH may be an efficacious therapy for men with idiopathic osteoporosis. PMID- 11730262 TI - Treatment of osteoporosis in men with fluoride alone or in combination with bisphosphonates. AB - Alendronate has been approved as a first antiresorptive drug for treatment of osteoporosis in men. Except for fluoride, in some countries so far there has been no approved anabolic substance for the treatment of male osteoporosis. From small studies in men and male patients included in studies on postmenopausal osteoporosis there is sufficient evidence that fluoride has the same osteoblast stimulating potency in men and women. In our own study on 64 men with idiopathic osteoporosis without prevalent fractures, a low-dose intermittent fluoride regimen (15 mg fluoride ions 3 months on, 1 month off) resulted in an average gain of lumbar spine BMD of 3% per year and a lower rate of incident fractures as compared with patients treated with calcium only. A combination of fluoride with an antiresorptive drug may improve the therapeutic results in terms of pattern of biochemical marker response and gain in BMD. This was shown for postemopausal osteoporosis in several studies using fluoride and hormone replacement therapy (HRT). Encouraged by a Dutch study using etidronate/fluoride in corticoid-induced osteoporosis, we performed a pilot study in 33 men with severe established primary osteoporosis giving cyclically etidronate for 14 days followed by fluoride plus calcium/vitamin D for 76 days. This combined regimen resulted in significantly higher increases of BMD than fluoride or etidronate alone. In an ongoing trial we are studying a continuous, combined treatment of alendronate and fluoride plus calcium/vitamin D in established idiopathic osteoporosis in men. The results of a preliminary evaluation look very promising. A large study with a bisphosphonate plus fluoride, taking fractures as the primary endpoint and bone biopsies to assess safety, would be very valuable. PMID- 11730263 TI - Cell volume regulatory mechanisms in progression of renal disease. AB - One of the striking morphological features of renal failure is an increase of cell volume. This review explores the role of cell volume regulatory mechanisms in the pathophysiology of progressive renal disease. The case is made that TGF beta, a major cytokine involved in the development of progressive renal failure, upregulates the transcription of the serum and glucocorticoid-dependent kinase hSGK1, involved in cell volume regulation. Excessive extracellular glucose concentrations stimulate TGF-beta1 expression and thus similarly enhance hSGK1 transcription. The kinase stimulates two mechanisms important for cell volume regulation, i.e. the renal epithelial Na+ channel ENaC and the thick ascending limb Na+,K+,2Cl- cotransporter BSC1. On the one hand, stimulation of renal tubular transport leads to renal retention of Na+, which favours the development of hypertension. On the other, the increase of cell volume stimulates protein synthesis and inhibits protein degradation, contributing to the enhanced net formation and deposition of matrix proteins. At later stages, the increase of cell volume may be reversed to atrophy, and cell death may lead to loss of functional tissue. In conclusion, progressive renal disease is paralleled by deranged cell volume regulatory mechanisms. PMID- 11730264 TI - Intrauterine programming of nephron number: the fetal flaw revisited. AB - A broad range of epidemiological evidence supports the hypothesis that risk of essential hypertension, coronary heart disease and non-insulin dependent diabetes is, in part, determined before birth. This phenomenon, termed programming, is now the subject of intensive investigation in order to determine possible underlying mechanisms. It is widely accepted that maternal nutritional status in pregnancy is a major programming influence upon the fetus. This review considers the hypothesis that nephron number in humans is determined by prenatal nutrition. An increasing number of human studies indicate that the developing kidney is particularly vulnerable to the adverse effects of fetal growth retarding influences. In animals, growth retarding diets or other insults which have an impact upon the development of cardiovascular functions, also appear to impact upon nephron number. However, it is possible that hypertension and reduced renal reserve merely coincide and are not causally associated. PMID- 11730265 TI - Pharmacological intervention in renal fibrosis and vascular sclerosis. AB - Progressive renal disease is associated with the development of fibrosing lesions not only in the glomerulus, but also in the interstitial and vascular compartments of the kidney. A growing body of work suggests that the mechanisms involved in this process are to a large extent shared by the glomerular mesangial cell, tubulointerstitial fibroblast and vascular smooth muscle cell. In this review we consider evidence that treatment strategies focused on any one of these cells are likely to be of universal benefit in the abrogation of the ongoing scarring that accompanies progressive renal disease, while at the same time reducing the progressive vascular sclerosis so often ultimately responsible for the excessive mortality seen in patients with renal failure. PMID- 11730266 TI - Epidemiology of primary glomerulonephritis in the elderly. Report from the Italian Registry of Renal Biopsy. AB - BACKGROUND: In the pre-biopsy era, primary glomerulonephritis (GN) was thought to pertain to children and adults. On the contrary, as renal biopsy came into clinical practice, this disease was recognized to affect the elderly too, even if the rate of such an occurrence still had to be defined. Recently, the Italian Registry of Renal Biopsy provided the opportunity to ascertain the diffusion of elderly GN in epidemiological terms. METHODS: Eighty-eight per cent of the Italian Renal Units known to routinely perform renal biopsy reported the records of patients who underwent biopsy investigation in 1996. Thus, 2,511 cases were ranked according to patient's age, histological diagnosis and clinical manifestation. The age-related incidence of each clinicopathological picture was calculated as the ratio between the number of patients found to be affected during the year and the number of subjects at risk, namely all age-matched people living in Italy in the same year. RESULTS: A diagnosis of primary GN was found in 1,474 records. Elderly patients accounted for 19% of those biopsy-proven primary GN, though elderly people were only 16% of the Italian population. This disproportion indicated that the incidence of primary GN in the elderly was higher than expected: actually, when compared to the incidence in the adult, it was 30.8 vs. 28.3 cases per million of population (PMP). Four histological pictures were found to be more frequent in the elderly than in the adult: membranous GN (13.4 vs. 4.2 PMP), crescentic GN (3.1 vs. 0.9 PMP), membranoproliferative GN (2.9 vs. 0.4 PMP), acute post-streptococcal GN (0.9 vs. 0.4 PMP). As for clinical manifestations, all GN-related symptoms were more frequent in the elderly than in the adult, except one: urinary abnormalities (4.5 vs.14.1 PMP). CONCLUSION: Through the Italian Registry of Renal Biopsy, primary GN was found to occur more frequently in the elderly than in the adult. Such a high incidence would be even higher if aged patients showing urinary abnormalities regularly underwent renal biopsy. As this result reverses all previous views, many issues about elderly GN should be reconsidered. PMID- 11730267 TI - Hospitalizations for bacterial endocarditis after renal transplantation in the United States. AB - PURPOSE: The national rate of and risk factors for bacterial endocarditis in renal transplant recipients has not been reported. METHODS: Retrospective registry study of 33,479 renal transplant recipients in the United States Renal Data System (USRDS) between 1 July 1994 and 30 June 1997. Hospitalizations for a primary diagnosis of bacterial endocarditis (ICD-9 codes 421.x) within three years after renal transplant were assessed. RESULTS: Renal transplant recipients had an unadjusted incidence ratio for endocarditis of 7.84 (95% confidence interval 4.72-13.25) in 1996. In multivariate analysis, a history of hospitalization for valvular heart disease (adjusted odds ratio (AOR), 25.81, 95% confidence interval 11.28-59.07), graft loss (AOR, 2.81, 95% CI 1.34-5.09), and increased duration of dialysis prior to transplantation were independently associated with hospitalizations for bacterial endocarditis after transplantation. Hospitalization for endocarditis was associated with increased patient mortality in Cox Regression analysis, hazard ratio 4.79, 95% CI 2.97 6.76. CONCLUSIONS: The overall incidence of bacterial endocarditis was much greater in renal transplant recipients than in the general population, although it is still relatively infrequent. Independent risk factors for bacterial endocarditis in the renal transplant recipients were identified, the most significant of which was valvular heart disease. Endocarditis substantially impacts renal transplant recipient survival. PMID- 11730268 TI - Risk factors for hospitalizations resulting from pulmonary embolism after renal transplantation in the United States. AB - BACKGROUND: Risk factors for pulmonary embolism (PE) have been identified in the general population but have not been studied in a national population of renal transplant recipients. METHODS: Therefore, 33,479 renal transplant recipients in the United States Renal Data System from 1 July 1994-30 June 1997 were analyzed in a historical cohort study of hospitalized PE (ICD9 Code 415.1x). HCFA form 2728 was used for comorbidities. RESULTS: Renal transplant recipients had an incidence of PE of 2.26 hospitalizations per 1000 patient years at risk. In multivariate analysis, polycystic kidney disease (adjusted odds ratio, 4.44, 95% confidence interval, 2.31-8.53), older recipient age, higher recipient weight, cadaveric donation, history of ischemic heart disease, and decreased serum albumin were associated with increased risk of PE. Body mass index and hemoglobin were not significant. Kidney-pancreas transplantation was also not significant. In Cox Regression analysis PE was associated with increased mortality (hazard ratio 2.06, 95% CI 1.34-3.18). CONCLUSIONS: The most important risk factors for PE in this population were polycystic kidney disease, advanced age and increased weight. The reasons for the increased risk of polycystic kidney disease remain to be determined but were independent of hematocrit level at initiation of end stage renal disease, and may result from venous compression. Prospective studies of anatomical and hemostatic changes after renal transplantation in recipients with polycystic kidney disease are warranted. PMID- 11730269 TI - The impact of renal transplantation on the incidence of congestive heart failure in patients with end-stage renal disease due to diabetes. AB - BACKGROUND: Patients with end stage renal disease (ESRD) are at increased risk for cardiovascular disease. We hypothesized that the clinical incidence of congestive heart failure (CHF) would be lessened after successful renal transplantation, as many of the metabolic and intravascular volume abnormalities associated with dialysis-dependent ESRD would resolve. METHODS: Using data from the USRDS, we studied 11,369 patients with ESRD due to diabetes enrolled on the renal and renal-pancreas transplant waiting list from 1 July 1994-30 June 1997. Cox non-proportional hazards regression models were used to calculate adjusted, time-dependent hazard ratios (HR) for time to the most recent hospitalization for CHF (including acute myocardial infarction, unstable angina, or other CHF, ICD9 Code 428.x) for a given patient in the study period, controlling for both demographics and comorbidities in the medical evidence form (HCFA 2728). RESULTS: In comparison to maintenance dialysis, renal transplantation was independently associated with a lower risk for CHF (HR 0.64, 95% confidence interval, 0.54 0.77) in a model including age, gender, race, and year of first dialysis, but not in a model including comorbidities from the medical evidence form, although the sample was much smaller. CONCLUSIONS: Patients with ESRD due to diabetes on the renal transplant waiting list were much less likely to be hospitalized for congestive heart failure after renal transplantation, despite post transplant complications due to immunosuppression. PMID- 11730270 TI - Human immunodeficiency virus/acquired immunodeficiency syndrome-associated nephropathy at end-stage renal disease in the United States: patient characteristics and survival in the pre highly active antiretroviral therapy era. AB - BACKGROUND: The patient characteristics and course of HlV/AIDS-associated nephropathy (HIVAN) are presented for a national sample of end-stage renal disease (ESRD). METHODS: 375,152 patients in the United States Renal Data System were initiated on ESRD therapy between 1 January 1992 and 30 June 1997 and analyzed in an historical cohort study of HIVAN. RESULTS: Of the study population, 3653 (0.97%) had HIVAN. Among patients with HIVAN, 87.8% were African American. HIVAN had the strongest association with African American race compared to other causes of renal failure except sickle cell anemia in logistic regression analysis (odds ratio 12.20, 95% confidence interval (CI) 10.57-14.07). In a separate logistic regression analysis, HIVAN was associated with male gender, decreased age (39.32 +/- 8.51 vs. 60.97 +/- 16.43 years, p<0.01 by Student's t test), weight, body mass index, hemoglobin, albumin, decreased rate of pre dialysis erythropoietin use, increased creatinine, decreased hypertension and increased rate of no medical insurance. The geographic distribution of HIVAN was similar to the distribution of HIV cases nationally. Two-year all cause unadjusted survival was 36% for HIVAN vs. 64% for all other patients with ESRD. HIVAN was associated with decreased patient survival in Cox regression analysis (hazard ratio for mortality 5.74, 95% CI, 5.40-6.10). CONCLUSIONS: HIVAN had the strongest association with African American race of all causes of renal failure among patients on maintenance dialysis. HIVAN was associated with decreased patient survival after initiation of dialysis, which may be associated with poorer medical condition at initiation of dialysis. PMID- 11730271 TI - HIV nephropathy and the Duffy antigen/receptor for Chemokines in African Americans. AB - BACKGROUND AND OBJECTIVES: HIV nephropathy (HIVAN) is markedly racially biased in its distribution, occurring in about 10% of HIV infected African Americans according to some studies. Based upon previous laboratory and epidemiological studies, the Duffy promoter polymorphism, which occurs almost exclusively in individuals of African descent, has been postulated to be the predisposing factor. We aimed to explore that relationship by directly genotyping individuals with HIV nephropathy to determine the proportion homozygous for this mutation to test the hypothesis it was responsible for the genetic component of this disease. We anticipated that if the polymorphism was associated with HIV nephropathy all individuals would be homozygous for this mutation. METHOD: Individuals with HIVAN proven on biopsy were identified from previous studies and a pre-existing clinical database. This diagnosis was confirmed by an experienced pathologist examining the biopsies in a blinded fashion. PCR and RFLP strategies were used on the biopsy samples to genotype for the Duffy promoter polymorphism. The cases were compared to a control population of HIV seronegative African Americans. RESULTS: Twenty African American individuals with HIV nephropathy were successfully genotyped. Only nine were homozygous for the promoter mutation. Nine were heterozygous and two homozygous wild type. Furthermore, the frequency of the polymorphism did not differ from the background rate in the African American population (OR = 0.788 95% confidence intervals 0.378-1.64). CONCLUSION: The Duffy promoter polymorphism was not disproportionately represented in persons with HIVAN calling into question any significant role in the pathogenesis of HIVAN. PMID- 11730272 TI - Prevalence of serum antibodies to hepatitis B and C viruses in patients with primary glomerulonephritis. AB - We retrospectively analysed the medical records of patients who underwent renal biopsy in our nephrology department over the past 8 years. Our aim was to discover the frequency of hepatitis B (HBV) and hepatitis C virus (HCV) in patients with primary glomerulonephritis (GN) in Turkey, a moderately endemic country for both infections. The study included 144 male and 104 female patients aged between 12-70 years. The frequency of HBsAg and anti-HCV were found to be similar to healthy controls, respectively 6.5% and 1.6% (p>0.05). As is true for low endemic countries, HBV and HCV infections might be rarely associated with GN in Turkey. PMID- 11730273 TI - Molecular analysis of uromodulin and SAH genes, positional candidates for autosomal dominant medullary cystic kidney disease linked to 16p12. AB - BACKGROUND: The location of a second genetic locus for autosomal dominant medullary cystic kidney disease (ADMCKD) at chromosome 16p12 led us to further investigate the molecular analysis of the critical region where two genes coding for uromodulin and SA proteins with renal specific functions, UMOD and SAH, are localized. METHODS: We characterized the intron-exon boundary sequences by screening phage and BAC DNA genomic clones for the development of new molecular tools functional to the mutation analysis of UMOD and SAH genes. RESULTS: No consistent mutations for ADMCKD2 were found in the UMOD and SAH genes. We identified a silent polymorphism in the UMOD gene at codon C174 which co segregates with the disease in the ADMCKD2 family. CONCLUSIONS: This study excludes the involvement of uromodulin and SAH genes in ADMCKD2, and provides new tools for their molecular analysis in other diseases. PMID- 11730275 TI - Behavioural stress blunts the creatinine clearance increase induced by a protein load in healthy subjects. AB - BACKGROUND: The aim of this study was to investigate how behavioural states related to different levels of stress affected the increments of glomerular filtration rate induced by an acute protein load. METHODS: Thirteen healthy subjects were enrolled. Each subject was studied from 9:00 h to 15:00 h on two consecutive days. In random order, after a protein meal (1.2 g/kg b.w. of protein), each subject was required to remain in a relaxing, sitting position (resting period, R), or to solve graphical and mathematical problems (behavioural stress period, S). Mean blood pressure (MBP) and heart rate (HR) were monitored by an ambulatory blood pressure device. Urine samples collected in each period were used to measure glomerular filtration rate (GFR, creatinine clearance) and urinary sodium excretion (UNa+V). RESULTS: Significant decreases in MBP and HR were observed during the resting period after the protein load, which significantly increased GFR. There was also a large increase of UNa+V. During S, the GFR changes were no longer seen whereas the increse of UNa+V was maintained. HR and MBP did not change compared to the prestimulus period. CONCLUSIONS: We conclude that in healthy subjects behavioural stress can blunt the increase in GFR that follows a protein load, presumably by neuro-humoral activated mechanisms. In our experimental conditions, behavioural stress did not affect UNa+V. PMID- 11730274 TI - Low dose dopamine infusion reduces renal tubular injury following cardiopulmonary bypass surgery. AB - BACKGROUND: The use of dopamine to protect the kidneys against hypoperfusion injury remains controversial with little clinical evidence of benefit and increasing concerns regarding safety. In this double-blind, prospective, randomised study, we investigated the effect of dopamine infusion (2.5 microg/kg/min) on glomerular filtration rate (GFR) and tubular injury in patients undergoing routine cardiopulmonary bypass (CPB). METHODS: Forty eight patients were randomly assigned to receive intravenous dopamine or saline from induction of anaesthesia until 48 hours post-operatively. There were no differences in mean age, bypass time or pre-op creatinine in the 36 patients (33 men) who completed the study. 51Cr-EDTA GFR (ml/min/1.73 m2) was measured pre-operatively and on day 5 only. Urinary markers of tubular injury (albumin, N-acetyl glucosaminidase, NAG; retinol binding protein, RBP) were measured pre-operatively, and on days 1, 2 and 5. RESULTS: GFR was preserved equally in both groups. All patients demonstrated significant tubular injury but urinary levels of NAG and RBP were lower in the dopamine group (41%, p=0.057 and 41%, p=0.007, respectively) on the first post-operative day. CONCLUSION: We conclude that low dose dopamine infusion may reduce renal tubular injury following CPB in patients with normal or near normal baseline renal function. PMID- 11730276 TI - Life threatening hyperkalemia and acidosis secondary to trimethoprim sulfamethoxazole treatment. AB - We present a 77-year-old male with moderate chronic renal insufficiency from diabetic nephropathy who developed severe metabolic acidosis and life threatening hyperkalemia on treatment with regular dose of trimethoprim-sulfamethoxazole (TMP SMZ) for urinary tract infection. The metabolic acidosis and hyperkalemia resolved upon appropriate medical intervention and discontinuation of TMP-SMZ. While hyperkalemia has commonly been reported with high dose of TMP-SMZ, severe metabolic acidosis is quite uncommon with regular dose TMP-SMZ. We emphasize that patients with renal tubular acidosis (RTA), renal insufficiency, aldosterone deficiency, old age with reduced renal mass and function, and angiotensin converting enzyme (ACE)-inhibitor therapy are at high risk of developing these severe and potentially life threatening complications. PMID- 11730277 TI - Maxillary brown tumor in secondary hyperparathyroidism requiring urgent parathyroidectomy. AB - Brown tumors are unusual but serious complications of renal osteodystrophy, and can be successfully treated by parathyroidectomy or by pharmacological treatment of hyperparathyroidism. Brown tumors in patients with severe hyperparathyroidism (HPT) secondary to renal failure have been increasingly reported. We describe an unusual case of brown tumors at the maxillary bone and the seventh right rib, in a 57-year old man with a long history of hemodialysis. The maxillary lesion caused serious local discomfort due to its rapid growth. In this setting, surgical total parathyroidectomy was chosen as the most adequate therapeutic approach, given the previous unsatisfactory response to calcitriol. After successful parathyroidectomy, rapid healing was achieved with sclerosis of both brown tumors, as documented by serial computerized tomograms. In conclusion, although vitamin D therapy has been beneficial in several cases of secondary hyperparathyroidism complicated by brown tumors, we propose that whenever regression of the tumor bulk is urgently needed, as in our case, parathyroidectomy should be the first treatment choice. PMID- 11730278 TI - Anterior ischemic optic neuropathy and dialysis: role of hypotension and anemia. AB - The pathogenesis of anterior ischemic optic neuropathy (AION) primarily involves interference with the posterior ciliary artery blood supply to the prelaminar optic nerve. Uremic patients often have coexisting pathology such as hypotension (decreased blood delivery), or hypertension, atherosclerosis (increased resistance to blood supply), and anemia (low blood oxygen carrying capacity), predisposing them to AION. We describe a 49-year-old patient on dialysis for many years. He had long-standing hypotension, worsened during each dialysis treatment. He awoke one morning at age 48 complaining of blurred vision in the left inferior field. Based on the clinical course, funduscopic and fluorangiographic examination and visual field defects, AION was diagnosed. Nine months after the loss of vision in the left eye, vision in the right eye became blurred and worsened over the next 24 hours. The diagnosis of AION in the right eye was made. At the last examination ten months later, the patient, still amaurotic, was given a very poor prognosis for further recovery of the visual defects. Surprisingly, very few cases of AION have been reported in chronic uremic patients on dialysis: to the best of our knowledge, only 12 including ours. Most of these cases share some features, including hypotension above all and anemia as common risk factors. Neither the type of dialysis treatment (hemo-, peritoneal dialysis) nor sex seem to have any influence on the occurrence of AION. Uremic children can be affected. What is striking in the three published pediatric cases is that they all had polycystic kidney disease. Treatment of AION in all 12 cases consisted of a combination of steroids, i.v. saline, blood transfusions and rhEpo. AION was more frequently bilateral and irreversible, ending in permanent amaurosis. In conclusion, this study aims to stress that most cases of AION occurring in chronic uremic patients on dialysis have some common features, including hypotension above all and anemia as common risk factors. PMID- 11730279 TI - Reversible MRI changes in a patient with uremic encephalopathy. AB - A 19-year-old patient on chronic ambulatory peritoneal dialysis experienced severe neurologic disturbances caused by uremia. Increased signal intensity was seen bilaterally in the cortical and subcortical areas of the occipital and parietal lobe on cranial magnetic resonance imaging (MRI). Insufficient peritoneal dialysis efficacy was documented and the patient was switched from peritoneal to hemodialysis. Cranial MRI indicated a marked regression of the lesions to nearly normal, confirming the diagnosis of uremic encephalopathy. PMID- 11730280 TI - HBV-related cutaneous periarteritis nodosa in a patient 16 years after renal transplantation: efficacy of lamivudine. AB - Cutaneous periarteritis nodosa (PAN) is a clinical feature characterized by chronic, benign course; its pathogenesis is unknown. In patients submitted to renal transplantation cutaneous PAN is a rare complication. We report a case of cutaneous PAN associated with the reappearance of hepatitis B antigen 16 years after kidney transplantation. A 44-year-old man underwent successful renal transplantation in June 1980. In December 1996 he presented multiple painful erythematous subcutaneous nodules on both legs. Skin lesion biopsy showed the presence of cutaneous PAN. Six months later laboratory data demonstrated the presence of HbsAg. HBeAg, HBcAb and detectable HBV-DNA serum by polymerase-chain reaction (PCR) assay. Anti-HBs and anti-HBe proved negative. In July 1998 the laboratory tests showed an important increase of HBV-DNA (5.1 billion by Branched DNA), and so lamivudine (100 mg/day) was introduced. HBV-DNA became undetectable by PCR after 3 months of therapy. Seven months later a new skin biopsy was performed. The typical signs of PAN were no longer evident. As HBV infecion was demonstrated six months after the clinical appearance of the PAN, in a patient who was believed to be immune to the virus, it is possible that, in the early stages, the hepatitis B antigen title was methodologically indeterminable, but sufficient to form circulating immune complexes responsible for vasculitis primer. Lamivudine therapy resulted efficacious in favouring the regression of cutaneous PAN, but its long-term efficacy requires further evaluation as regards potential selection of drug resistant hepatitis B virus (HBV) mutants during treatment. PMID- 11730281 TI - Ciprofloxacin-associated Achilles tendon rupture in a hemodialysis patient. PMID- 11730282 TI - Addressing curriculum skills development. PMID- 11730283 TI - Make your voices known for peace. PMID- 11730284 TI - Everything changed on September 11th. PMID- 11730285 TI - Piffalls of precepting. PMID- 11730286 TI - A senior elective: promoting health in underserved communities. AB - OBJECTIVES: A fourth-year service-learning elective was developed to teach medical students about the social, economic, and culturalfactors that affect health and health-seeking behavior DESCRIPTION: The elective provides students with didactic material and educational experiences in public housing sites in the community. Students work closely with community members to implement a community health care intervention. EVALUATION: Students respond to 10 reflection questions that assess their understanding of the important community factors that affect health. CONCLUSIONS: Reflection responses demonstrate that the course has challenged assumptions, dispelled stereotypes, and enhanced awareness of the role of socialfactors in maintaining health. PMID- 11730287 TI - What doesn't kill us makes us stronger. PMID- 11730288 TI - Imagine a woman by Richard Seizer: uses in a doctoring course. PMID- 11730289 TI - Longitudinal residency training: a survey of family practice residency programs. AB - BACKGROUND AND OBJECTIVES: Most family practice residency training consists of 2 4-week block rotations in specific curricular areas, supplemented by training in the family practice center. An alternative model, longitudinal residency training, emphasizes training in curricular areas over a 3-year time period. This study determined the frequency of longitudinal training in family practice residency programs. METHODS: We conducted a survey of 477 residency program directors listed in the American Academy of Family Physicians 2000 Directory of Family Practice Residency Programs. RESULTS: Sixty-eight percent (n=320) of program directors responded to the survey. A total of 3.6% of program directors described their program as "mostly longitudinal," and 14.2% described their program as "half block/half longitudinal." An additional 15% of program directors indicated interest in adopting or moving toward a longitudinal program in the next 2 wears. Responses suggest some inconsistencies in program directors' understanding of what constitutes a longitudinal curriculum. CONCLUSIONS: Longitudinal residency training is reported in 18% of family practice residency programs. Further work is needed to develop a definition of longitudinal residency training. PMID- 11730290 TI - Longitudinal versus traditional residencies: a study of continuity of care. AB - BACKGROUND AND OBJECTIVES: Continuity of care is one of the presumed advantages of longitudinal residencies. However, it is not clear how well such residencies provide continuity of care, and, further, there is no recognized acceptable rate of good continuity. We compared traditional and longitudinal residencies to determine the extent to which the residents provided their patients with continuity of care. METHODS: We conducted a systematic chart review at three longitudinal and three matched traditional block-rotation programs. In total, 628 charts were reviewed, and 6,256 visits were evaluated. Continuity with a primary resident was evaluated over a 2-year period, with continuity defined as the percentage of visits for which the patient saw the same resident. RESULTS: There was no significant difference in overall rates of continuity between longitudinal and traditional programs (59.6% versus 57.8%). One longitudinal program, however, had a 74.8% rate of continuity, which was significantly higher than the rates in the otherfive programs. CONCLUSIONS: There was no significant difference found in continuity of care provided by residents at longitudinal programs, compared with those at traditional programs. Our results do not support the hypothesis that longitudinal residency programs achieve superior rates of continuity of care. Further comparison studies of longitudinal and traditional programs would be useful. PMID- 11730291 TI - Improving continuity by increasing clinic frequency in a residency setting. AB - BACKGROUND AND OBJECTIVES: Continuity of care is required in family practice training programs. However, continuity for some patients may not be adequately served in the traditional training model that has residents in the family practice center (FPC) for 1 to 3 half-day clinics per week. This study sought to determine if increasing clinic frequency in a family practice residency has an effect on continuity of care. METHODS: On January 1, 1999, the residency program changedfrom a traditional clinic scheduling model to one where all residents saw patients in the FPC 4 to 5 days a week. By using shorter clinic sessions, total resident time in the FPC was nearly unchanged (decreasing 5% overall). We reviewed 1,709 randomly selected billing records for residents' patients who frequently utilized medical care (three or more visits within 6 months) and assessed continuity for 1 year before and after this intervention, using both the modified, modified continuity index (MMCI) and the percentage of visits to the primary care provider (PCP). RESULTS: Overall, the MMCIfor patients who frequently saw residents increasedfrom .59 to .64. The average frequency with which these patients saw their PCPs improvedfor the first-year class (from 51% of visits before implementation to 72% after) and the third-year class (from 66% of visits to 72%). CONCLUSIONS: Scheduling daily resident clinics in the FPC increased continuity among patients who frequently saw residents beyond that achieved using traditional scheduling, without increasing total resident time in the FPC. PMID- 11730292 TI - Are some subjects better taught in block rotation? A geriatric experience. AB - BACKGROUND AND OBJECTIVES: This study characterizes the progress in and effectiveness of learning geriatric medicine during longitudinal and block phases of a family practice residency program. METHODS: A structured second-year geriatric block rotation was added to a residency longitudinal curriculum. To assess learning, a Geriatric Assessment Instrument (GAI) consisting of 50 multiple choice questions was administered to three classes offamily practice residents (n=33)five times during training: entry into the program, beginning of the second year, pre- andpost-geriatric block rotation, and at graduation. Improvement between individual residentfirst- and third-year in-training exam scores in geriatrics of the intervention classes were compared with the four classes that preceded the introduction of the block rotation (n=38). RESULTS: Scores on the GAI improved significantly before and after the rotation but not during any other interval of training during the residency. In-training exam scores improved significantlyfor the classes taught with the block rotation over those without it. CONCLUSIONS: Most of the geriatric learning occurred during the 1-month geriatric block rotation during the residency. In-training geriatric exam scores improved significantly with a geriatric block rotation. The use of structured repetitive learning experiences during the rotation to emphasize the common clinical issues and the skewed exposure to geriatric patients in the random nature of residency clinic and inpatient encounters accountfor this result. PMID- 11730293 TI - Longitudinal residency training in family medicine: not ready for prime time. AB - Many family medicine educators have called for changing the family practice residency curriculum from a series of block rotations to a longitudinal curriculum. A longitudinal curriculum is one in which residents are based in the family practice center every day or nearly every day of all 3 years of their residency training. Residents learn most of the clinical content offamily medicine through experiences with patients from their continuity clinics, under supervision of family medicine faculty, rather than through specialty-specific block rotations supervised by specialists. An important purported benefit of longitudinal training is improved continuity of care between residents and their patients. Unfortunately, definitions of longitudinal training vary widely, and at least one study shows that supposedly longitudinal curricula do not result in better continuity of care. Further, there is some evidence that acquisition of knowledge by residents may be better with intensive block rotations than with longitudinal training. Thus, the supposed benefits of longitudinal residency training remain unproven. PMID- 11730294 TI - Rural-urban and gender differences in procedures performed by family practice residency graduates. AB - OBJECTIVE: We compared the types of procedures performed and obstetrical care provided by family practice residency graduates, by practice location and physician gender METHODS: We conducted a cross-sectional questionnaire survey of 702 graduates who completed family practice residency programs in Alberta, Canada, from 1985 to 1995, inclusive. Graduates were asked to indicate which of 28 procedures and 7 obstetrical care practices they performed. The data were analyzed by gender and current practice location. RESULTS: A total of 442 (63%) of the graduates responded to the survey. The top five procedures performed by family practice graduates were minor office surgery, foreign body removal (eye), joint aspiration, joint injection, and anterior nasal packing. There was a declining trend in the number of procedures performed by family practice graduates from rural, to regional, to metropolitan areas. Relatively more males performed procedures; however, more females did IUD insertion and obstetrical care practice. Except for a few exceptions, a similar proportion of male and female graduates in rural practice performed procedures. CONCLUSIONS: The procedural and obstetrical care pattern of practice differs between family practice graduates in rural and urban areas, as well as between male andfemale graduates. Family practice residency programs should consider additional training in procedural skills for those planning to practice in rural areas, as well as encourage females to become skilled at performing procedures relevant to family practice. PMID- 11730295 TI - Development of the HIV/AIDS Q-sort instrument to measure physician attitudes. AB - BACKGROUND: Providers'attitudes about HIV/AIDS are an important dimension in the delivery of quality care to persons with HIV/AIDS. It is believed that education can alter attitudes, but there is a needfor a user-friendly instrument to measure the effect that HIV/AIDS educational programs have on attitudes. METHODS: A pool of HIV/AIDS attitude descriptors was collected through literature review and from individuals working in the HIV/AIDS field. Out of this pool of 90 descriptors, 48 descriptors with the highest face validity were selected through expert consensus ranking to create a preliminary survey instrument. Twenty-six physicians completed a pilot Q-Sort instrument with 48 descriptors. A variance analysis was conducted, and the top 28 descriptors with the most variability were selected for the final Q-Sort instrument, which was then completed by 191 physicians. A factor analysis was conducted to identify a small number of factors that explained the 28 descriptors. A subsample of 22 physicians repeated the test to establish test retest reliability. RESULTS: Factor analysis revealed three factors: (1) emotionality, (2) ability, and (3) reluctance. The Q-Sort instrument demonstrated good test-retest reliability, with reliability for the three factors of .82, .80, and .88, respectively. CONCLUSIONS: This Q-sort instrument is a reliable method for measuring physician attitudes toward HIV/AIDS patients. Further studies can test its use for evaluating the effect of educational programs on changing provider attitudes. PMID- 11730296 TI - Forgiveness. PMID- 11730297 TI - Laser vibrometric studies of sound-induced motion of the body walls and lungs of salamanders and lizards: implications for lung-based hearing. AB - A laser Doppler vibrometer was used to measure the acoustic responses of different body surfaces of several species of salamanders and lizards. The lateral body wall over the lung displayed sound-induced motion up to 30 dB greater than the lateral head surface from 300-1,000 Hz in salamanders and from 200-2,500 Hz in lizards. The lateral body wall of lungless plethodontid salamanders showed no such enhanced motion to sound. The lateral body wall of lizards was more responsive than their tympanum to sound frequencies below about 1,250-2,000 Hz. The frequency of the peak response of lizard body walls matched the resonant frequency of a Helmholtz resonator with the volume and dimensions of their lungs. In contrast, the frequency of peak response of salamander body walls was well below the resonant frequencies calculated for both Helmholtz resonators and closed tubes with the dimensions and volumes of their lungs. Nonetheless, filling the lungs with saline dramatically reduced the responsiveness of the lateral body walls of both the lunged salamanders and the lizards. As previously demonstrated in anuran amphibians, the lateral body wall and lungs of salamanders and lizards may function in sound reception, especially at relatively low frequencies. PMID- 11730298 TI - Detection of fruit- and flower-emitted volatiles by olfactory receptor neurons in the polyphagous fruit chafer Pachnoda marginata (Coleoptera: Cetoniinae). AB - Olfactory receptor neurons on the antennae of the African fruit chafer species Pachnoda marginata (Coleoptera: Scarabaeidae) were examined through extensive use of gas chromatography linked with electrophysiological recordings from single olfactory receptor neurons. Contacted neurons were stimulated with a large number of extracted volatiles from 22 different fruits and with 64 synthetic plant compounds. Extracted fruit volatiles were identified using linked gas chromatography-mass spectrometry. In total, 48 different odor compounds were found to elicit responses. Analysis of the response spectra of the contacted neurons (n = 232) revealed the presence of 28 classes of receptor neurons. The neurons exhibited strong selectivity as well as high sensitivity. Eleven of the identified classes were selectively activated by single compounds, while the remaining were activated by 2-6 compounds. Several receptor neurons that were activated by more than one compound responded to compounds sharing basic structural similarities. The results support the growing hypothesis that a significant proportion of plant-odor receptor neurons in insects are highly sensitive and selective for single odors. PMID- 11730299 TI - Echolocation by the barbastelle bat, Barbastella barbastellus. AB - When searching for insects along edges, Barbastella barbastellus alternated between two signal types. Type-2 signals had durations around 6 ms and were composed of an initial shallowly downward frequency modulated component, starting at about 45 kHz and followed by a shorter more steeply modulated component that ended at about 32 kHz. Type-1 signals were rather stereotyped with durations around 2.5 ms and a very short rise time. They covered an approximately 8 kHz wide frequency band positioned just below the 12-15 kHz-wide frequency band of type-2 signals, with no or small frequency overlap. In the recordings, type-1 signals almost had always a higher amplitude than type-2 signals, at least partly caused by head movements. Assuming that signal structure reflects function, we hypothesize that type-2 signals have the same adaptive value as the signals with a broadband and narrowband component of other vespertilionids, but with a reverse arrangement of the signal elements. Like the broadband component of the type-2 signals, type-1 signals are well suited to localize background targets. Thus, the localization component may be distributed among two signals separated in time, which has the advantage that both signals can be varied independently in the direction of emission and in amplitude. PMID- 11730300 TI - Stridulatory pattern generation in acridid grasshoppers: metathoracic interneurons in Stenobothrus rubicundus (Germar 1817). AB - Stridulation was elicited in tethered gomphocerine grasshoppers of the species Stenobothrus rubicundus in order to identify interneurons of the stridulation pattern generator, and describe their morphological and physiological properties. Nine types of such neurons could be found and characterized; eight of those could additionally be compared to corresponding neuron types previously known from other species. As shown in detail for one selected type, the neurons of the stridulation pattern generator are very similar in their anatomical appearance, and possess similar physiological qualities at least in two species with similar stridulation patterns. Stridulation interneurons of species with largely different stridulatory motor patterns have a similar morphology, but show a different activation timing throughout the stridulation. Nevertheless, special properties such as resetting or initiation capability of certain stridulation interneurons seem to be conserved throughout the species. The results suggest that the stridulation pattern generator of different species consists of a uniform set of interneurons that change their activity pattern to produce species specific song movements. PMID- 11730301 TI - Cannibalism and early capping: strategy of honeybee colonies in times of experimental pollen shortages. AB - We observed the impact of bad pollen supply (non-foraging due to artificial rain and pollen removal under poor-foraging conditions) on the survival of honey bee larvae, and on the total development time from egg-laying to the capping of a larval cell. Five days of non-foraging led to cannibalism of larvae younger than 3 days old and to a shortening of the time until larvae were sealed, but 4- and 5 day-old larvae survived even worse pollen supply situations. Manual pollen removal and reduction of income (pollen trap) induced cannibalism of younger larvae. The larvae's mean capping age significantly correlated with the mean pollen income: the less pollen was stored by the hive during the larvae's development, the earlier the larvae were capped. Both behavioral patterns lead to a quick reduction in the amount of unsealed older brood in response to a shortage of available protein. Older larvae have the highest pollen demand, so this strategy compensates for a shortage of supply by reducing demand. Additionally worker jelly gets enriched by protein gained from cannibalism, and the early capping of older larvae saves the oldest part of the brood, which represents the highest broodcare investment. PMID- 11730302 TI - Population coding of motion patterns in the early visual system. AB - Using extracellular recordings and computational modeling, we study the responses of a population of turtle (Pseudemys scripta elegans) retinal ganglion cells to different motion patterns. The onset of motion of a bright bar is signaled by a rise of the population activity that occurs within less than 100 ms. Correspondingly, more complex stimulus movement patterns are reflected by rapid variations of the firing rate of the retinal ganglion cell population. This behavior is reproduced by a computational model that generates ganglion cell activity from the spatio-temporal stimulus pattern using a Wiener model complemented by a non-linear contrast gain control feedback loop responsible for the sharp transients in response to motion onset. This study demonstrates that contrast gain control strongly influences the temporal course of retinal population activity, and thereby plays a major role in the formation of a population code for stimulus movement patterns. PMID- 11730303 TI - Asymmetrical generalisation between pheromonal and floral odours in appetitive olfactory conditioning of the honey bee (Apis mellifera L.). AB - The capacity to generalise between similar but not identical olfactory stimuli is crucial for honey bees, allowing them to find rewarding food sources with varying volatile emissions. We studied bees' generalisation behaviour with odours having different biological values: typical floral odours or alarm compounds. Bees' behavioural and peripheral electrophysiological responses were investigated using a combined proboscis extension response conditioning-electroantennogram assay. Bees were conditioned to pure linalool (floral) or to pure isoamyl acetate (alarm) and were tested with different concentrations of both compounds. Electrophysiological responses were not influenced by conditioning, suggesting that the learning of individual compounds does not rely on modulations of peripheral sensitivity. Behaviourally, generalisation responses of bees conditioned to the alarm compound were much higher than those of bees conditioned to the floral odour. We further demonstrated such asymmetrical generalisation between alarm and floral odours by using differential conditioning procedures. Conditioning to alarm compounds (isoamyl acetate or 2-heptanone) consistently induced more generalisation than conditioning to floral compounds (linalool or phenylacetaldehyde). Interestingly, generalisation between the two alarm compounds, which are otherwise chemically different, was extremely high. These results are discussed in relation to the neural representation of compounds with different biological significance for bees. PMID- 11730304 TI - Sensory modification of leech swimming: interactions between ventral stretch receptors and swim-related neurons. AB - The neuronal circuits that generate the leech swimming rhythm comprise oscillatory interneurons that provide appropriately phased output to drive swim related motoneurons. Within ganglia, these interneurons express three phases; between ganglia there exists a phase delay between homologs. Our earlier experiments revealed that stretch receptors embedded in the body wall participate in intersegmental coordination and setting intersegmental phases. To identify the basis for these sensory effects, we mapped interactions between a ventral stretch receptor and swim-related neurons. Connections between this receptor and motoneurons are weak and variable in quiescent preparations, but during fictive swimming stretch receptor activation modulates motoneuron oscillations, hence, these effects are polysynaptic, mediated by interneurons. We identified a strong, nonrectifying, and apparently direct electrical connection between the stretch receptor and oscillator neuron 33. The ventral stretch receptor also interacts with most of the other oscillatory interneurons, including inhibitory inputs to cells 28 and 208, excitatory input to the contralateral cell 115, and mixed input to the ipsilateral cell 115. These direct and indirect interactions can account for previously described effects of body-wall stretch on motoneuron activity. They also could mediate the previously described modification of intersegmental phase relationships by appropriately phased stretch receptor activation. PMID- 11730305 TI - Uncoordinated growth of the primary care work force. AB - Family physicians, nurse practitioners and physician assistants are distinctly different in their clinical training, yet they function interdependently. Together, they represent a significant portion of the primary care work force. Training capacity for these professions has increased rapidly over the physician assistant decade, but almost no collaborative work force planning has occurred. PMID- 11730306 TI - Preventive strategies for chronic liver disease. PMID- 11730307 TI - Hypodermoclysis in the treatment of dehydration. PMID- 11730308 TI - FRAMES method helps assess patients' readiness to change. PMID- 11730309 TI - Vaginal examination using the Sims position. PMID- 11730310 TI - Preventive strategies in chronic liver disease: part I. Alcohol, vaccines, toxic medications and supplements, diet and exercise. AB - Chronic liver disease is the 10th leading cause of death in the United States. Hepatitis C virus infection is the most frequent cause of chronic liver disease and the most common indication for liver transplantation. Preventive care can significantly reduce the progression of liver disease. Alcohol and hepatitis C virus are synergistic in hastening the development of cirrhosis; therefore, patients with hepatitis C infection should abstain from alcohol use. Because superinfection with hepatitis A or B virus can lead to liver failure, vaccination is recommended. Potentially hepatotoxic medications should be used with caution in patients with chronic liver disease. In general, nonsteroidal anti inflammatory drugs should be avoided; acetaminophen in a dosage below 2 g per day is the safest choice. Many herbal remedies are potentially hepatotoxic, and only milk thistle can be used safely in patients who have chronic liver disease. Weight reduction and exercise can improve liver function in patients with fatty liver. PMID- 11730311 TI - Dermatomyositis. AB - Dermatomyositis is an idiopathic inflammatory myopathy with characteristic skin manifestations. Although the disorder is rare, with a prevalence of one to 10 cases per million in adults and one to 3.2 cases per million in children, early recognition and treatment are important ways to decrease the morbidity of systemic complications. An association with other connective tissue disorders (overlap syndrome) and malignancy make this diagnosis particularly important to primary care physicians. Patient management includes careful evaluation for underlying malignancy and liberal use of physical therapy, antihistamines, sunscreen and oral corticosteroids. Poor prognostic indicators include poorly responsive disease, delay in diagnosis and the presence of malignancy. The therapeutic goal is to maintain function and prevent or minimize sequelae. PMID- 11730312 TI - Hypodermoclysis: an alternative infusion technique. AB - Hypodermoclysis, the subcutaneous infusion of fluids, is a useful and easy hydration technique suitable for mildly to moderately dehydrated adult patients, especially the elderly. The method is considered safe and does not pose any serious complications. The most frequent adverse effect is mild subcutaneous edema that can be treated by local massage or systemic diuretics. Approximately 3 L can be given in a 24-hour period at two separate sites. Common infusion sites are the chest, abdomen, thighs and upper arms. The preferred solution is normal saline, but other solutions, such as half-normal saline, glucose with saline or 5 percent glucose, can also be used. Potassium chloride can be added to the solution bag if needed. Hyaluronidase can also be added to enhance fluid absorption. Hypodermoclysis can be administered at home by family members or a nurse; the technique should be familiar to every family physician. PMID- 11730313 TI - Disability certifications in adult workers: a practical approach. AB - Family physicians are frequently asked to complete disability certification forms for workers. The certification process can be contentious because of the number of stakeholders, the varying definitions of disability and the nature of the administrative systems. Insufficient training on disability during medical school and residency complicates this process. Disability systems discussed include workers' compensation, private disability insurance, the Americans with Disabilities Act and the Family and Medical Leave Act. Strategies that help the physician complete disability certification forms effectively include identification of disability type, ascertainment of the definition of disability being applied, evaluation of workplace demands and essential job functions, assessment of worker capacity, and accurate and timely completion of the forms in their entirety. PMID- 11730314 TI - Charcot foot: the diagnostic dilemma. AB - Primary care physicians involved in the management of patients with diabetes are likely to encounter the diagnostic and treatment challenges of pedal neuropathic joint disease, also known as Charcot foot. The acute Charcot foot is characterized by erythema, edema and elevated temperature of the foot that can clinically mimic cellulitis or gout. Plain film radiographic findings can be normal in the acute phase of Charcot foot. A diagnosis of Charcot syndrome should be considered in any neuropathic patient, even those with a minor increase of heat and swelling of the foot or ankle, especially after any injury. Early recognition of Charcot syndrome and immobilization (often with a total contact cast), even in the presence of normal radiographs, can minimize potential foot deformity, ulceration and loss of function. Orthopedic or podiatric foot and ankle specialists should be consulted when the disease process does not respond to treatment. PMID- 11730315 TI - Photo quiz. Unusual wrist bruise. PMID- 11730316 TI - Symptomatic treatment and H. pylori eradication therapy for nonulcer dyspepsia. PMID- 11730317 TI - PHS guidelines for management of occupational exposure to HBV, HCV and HIV: implementation for health care facilities. PMID- 11730318 TI - c-Mos and cyclin B/cdc2 connections during Xenopus oocyte maturation. AB - Fully-grown G2 arrested Xenopus oocytes can be induced to enter and progress into meiotic cell cycle by progesterone stimulation. This process is termed oocyte maturation. An early response to progesterone is the synthesis of the onco protein c-Mos, defined as the candidate initiator of Xenopus oocyte maturation, which triggers the MAPK cascade, MPF activation and promotes CSF activity. Here we review our current knowledge on the synthesis, activation and functions of c Mos in connection with MPF activation during maturation. We also discuss our recent results concerning the dispensability of cyclin B degradation in meiosis I meiosis II transition and the stabilization of c-Mos through its direct phosphorylation by cyclin B/cdc2. PMID- 11730319 TI - The pathway of MAP kinase mediation of CSF arrest in Xenopus oocytes. AB - A cytoplasmic activity in mature oocytes responsible for second meiotic metaphase arrest was identified over 30 years ago in amphibian oocytes. In Xenopus oocytes CSF activity is initiated by the progesterone-dependent synthesis of Mos, a MAPK kinase kinase, which activates the MAPK pathway. CSF arrest is mediated by a sole MAPK target, the protein kinase p90Rsk which leads to inhibition of cyclin B degradation by the anaphase-promoting complex. Rsk phosphorylates and activates the Bub1 protein kinase, which may cause metaphase arrest due to inhibition of the anaphase-promoting complex (APC) by a conserved mechanism defined genetically in yeast and mammalian cells. CSF arrest in vertebrate oocytes by p90Rsk provides a potential link between the MAPK pathway and the spindle assembly checkpoint in the cell cycle. PMID- 11730320 TI - From progesterone to active Cdc2 in Xenopus oocytes: a puzzling signalling pathway. AB - Since almost two decades, it is known that progesterone is responsible of the release of the prophase I arrest of amphibian oocytes and leads to the activation of the universal MPF, through a puzzling transduction pathway. It involves negative regulation of the cAMP-dependent protein kinase (PKA) and synthesis of new proteins, among them the c-Mos protooncogene product. The implication of the Mos/mitogenic activated protein kinase (MAP kinase) pathway in Cdc2 activation has been extensively studied and is now at the centre of a controversial debate. In this paper, we discuss the current progress and our recent results on the molecular mechanisms allowing progesterone to activate MPF and propose a model to partly resolve the long-standing inconsistencies concerning the role of Mos/MAP kinase during this process. PMID- 11730321 TI - Cell cycle regulation by p38 MAP kinases. PMID- 11730322 TI - Regulation of MAP kinases by docking domains. PMID- 11730323 TI - The Raf/MEK/ERK pathway: new concepts of activation. AB - The Raf/MEK/ERK signaling was the first MAP kinase cascade to be characterized. It is probably one of the most well known signal transduction pathways among biologists because of its implication in a wide variety of cellular functions as diverse -and occasionally contradictory- as cell proliferation, cell-cycle arrest, terminal differentiation and apoptosis. Discovery and understanding of this pathway have benefited from the combination of both genetic studies in worms and flies and biochemical studies in mammalian cells. However, ten years after, this field is still under debate and new molecular partners in the cascade continue to increase the complexity of its regulation. This review deals with the emergence of new concepts in the activation and regulation of the Raf/MEK/ERK module. In particular, the preponderant role of B-Raf is underlined, and the role of novel regulators such as KSR is discussed. PMID- 11730324 TI - MAP kinase dynamics in yeast. AB - MAP kinase pathways play key roles in cellular responses towards extracellular signals. In several cases, the three core kinases interact with a scaffold molecule, but the function of these scaffolds is poorly understood. They have been proposed to contribute to signal specificity, signal amplification, or subcellular localization of MAP kinases. Several MAP kinases translocate to the nucleus in response to their activation, suggesting that nuclear transport may provide a regulatory mechanism. Here we describe new applications for Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Loss In Photobleaching (FLIP), to study dynamic translocations of MAPKs between different subcellular compartments. We have used these methods to measure the nuclear/cytoplasmic dynamics of several yeast MAP kinases, and in particular to address the role of scaffold proteins for MAP-kinase signaling. PMID- 11730325 TI - Spatiotemporal regulation of the p42/p44 MAPK pathway. PMID- 11730326 TI - Plant MAP kinase pathways: how many and what for? AB - Mitogen activated protein kinases (MAPK) are important mediators in signal transmission, connecting the perception of external stimuli to cellular responses. MAPK cascades are involved in signalling various biotic and abiotic stresses, like wounding and pathogen infection, temperature stress or drought, but also some plant hormones, such as ethylene and auxin. Moreover, MAPKs have been implicated in cell cycle and developmental processes. In Arabidopsis mutant screens and in vivo assays several components of plant MAPK cascades have been identified. This review compares results obtained from functional analyses of MAPK cascades in plants with recent data obtained from searching the complete Arabidopsis genome. This analysis reveals that plants have an overall of 24 MAPK pathways of which only a small subset has been studied so far. PMID- 11730327 TI - RINGO efficiently triggers meiosis resumption in mouse oocytes and induces cell cycle arrest in embryos. AB - RINGO was identified as a Cdc2-binding and activating protein which is necessary and sufficient to trigger G2/M progression in Xenopus oocytes. We have investigated whether the function of RINGO is conserved in mouse oocytes. We show that RINGO induces Germinal Vesicle BreakDown (GBVD) in mouse oocytes. Mos is known to induce GVBD in mouse oocytes, and is also involved in the metaphase II arrest, which is due to the CSF (CytoStatic Factor) activity. We found that RINGO also has CSF activity and induces cleavage arrest after injection into one blastomere of a late two-cell mouse embryo, like Mos. However, RINGO also inhibits polar body extrusion of wild type mouse oocytes. The same effect of RINGO on first and second polar body extrusion was observed in Mos -/- mouse oocytes. The injection of RINGO mimics Mos effects: GVBD induction and efficient cleavage arrest. However, our results in mouse oocytes suggest that RINGO may have additional functions in meiosis regulation. PMID- 11730328 TI - Mobile bearings in primary knee arthroplasty. AB - Mobile-bearing knee arthroplasty (MBKA) has potential advantages compared with conventional fixed-bearing total knee arthroplasty (TKA). By allowing unconstrained axial rotation, MBKA can offer greater articular conformity without an increased probability of loosening due to increased axial torque. Increased articular conformity minimizes polyethylene contact stresses, thereby reducing linear wear and subsurface fatigue failure. Axial rotation of the platform also enables self-correction of tibial component malrotation. Despite these advantages, the long-term clinical results obtained with current MBKA devices are similar to those obtained with well-designed fixed-bearing TKA prostheses, with no data suggesting their superiority. The disadvantages of MBKA include bearing dislocation and breakage, soft-tissue impingement, a steep technique learning curve, and concerns about volumetric wear. Hypothetically, longer-term follow-up of MBKA results may reveal a significant difference from fixed-bearing TKA results as the fatigue failure threshold of incongruent polyethylene is exceeded. PMID- 11730329 TI - Recreational injuries in children: incidence and prevention. AB - Participation in eight common types of recreational activities leads annually to more than 2 million medically treated musculoskeletal injuries in children aged 5 to 14 years. Many of these injuries could have been prevented if current safety guidelines and protective equipment had been used. Studies have demonstrated the value of safety education programs in preventing injuries. Parents consider their child's physician an important source of safety education, and orthopaedic surgeons have a unique opportunity to provide injury prevention counseling. The American Academy of Orthopaedic Surgeons recognizes the importance of injury prevention and has developed advocacy programs that are readily available to physicians and the public. Individual orthopaedists should be involved in injury prevention through patient education, research, community programs, and regulatory efforts that promote safe play for children. PMID- 11730330 TI - Acute hand and wrist injuries in athletes: evaluation and management. AB - Acute hand and wrist injuries in the athlete constitute a unique orthopaedic challenge. Because of the particular demands on the athlete (e.g., financial implications, coaching and administration pressures, self-esteem issues), a specialized management approach is often necessary. Common sites of injury include the ulnar collateral ligament of the thumb metacarpophalangeal joint, proximal interphalangeal joint, metacarpals and phalanges, scaphoid, hamate, and distal radius. Treatment of these injuries varies depending on the patient's age, sport, position played, and level of competition, but departures from standard practice as regards surgery, rehabilitation, and return to competition should never compromise care. PMID- 11730331 TI - Developmental dysplasia of the hip from six months to four years of age. AB - Developmental dysplasia of the hip (DDH) denotes a wide spectrum of pathologic conditions, ranging from subtle acetabular dysplasia to irreducible hip dislocation. When DDH is recognized in the first 6 months of life, treatment with a Pavlik harness frequently results in an excellent outcome. In children older than 6 months, achieving a concentrically reduced hip while minimizing complications is more challenging. Bracing, traction, closed reduction, open reduction, and femoral or pelvic osteotomies are frequently used treatment modalities for children aged 6 months to 4 years. In the past, treatment recommendations have often been based on the patient's age. However, recent practice has placed more emphasis on addressing the specific disorder and avoiding iatrogenic osteonecrosis. The incidence of osteonecrosis of the femoral head has been reduced by avoiding immobilization of the hip in extreme abduction and by using femur-shortening osteotomies when appropriate. Pelvic osteotomy continues to gain favor for the treatment of selected patients over 18 months of age. PMID- 11730332 TI - Closed-chain rehabilitation for upper and lower extremities. AB - Closed-chain exercise protocols are used extensively in rehabilitation of knee injuries and are increasingly used in rehabilitation of shoulder injuries. They are felt to be preferable to other exercise programs in that they simulate normal physiologic and biomechanical functions, create little shear stress across injured or healing joints, and reproduce proprioceptive stimuli. Because of these advantages, they may be used early in rehabilitation and have been integral parts of "accelerated" rehabilitation programs. The authors review the important components of a closed-chain rehabilitation program and provide examples of specific exercises that are used for rehabilitation of knee and shoulder injuries. PMID- 11730333 TI - Characterization of noncovalent complexes of recombinant human monoclonal antibody and antigen using cation exchange, size exclusion chromatography, and BIAcore. AB - The binding of fully human monoclonal antibodies (MAbs) D2E7 and 2SD4 to their antigen, human tumor necrosis factor-alpha (TNFalpha), was investigated by BIAcore, cation exchange (CIEX), and size exclusion liquid chromatography (SEC) using ultraviolet and laser light scattering detectors. D2E7 has a higher affinity for TNFalpha than 2SD4 and the two antibodies (Abs) differ by 12 amino acids in the antigen (Ag) binding regions. A BIAcore biosensor instrument was used to determine the association, k(on) and dissociation, k(off), rate constants for the binding of TNFalpha to D2E7 and 2SD4. The HPLC methods were used to resolve and to study D2E7, 2SD4, and TNFalpha molecules and the noncovalent complexes of D2E7 and 2SD4 with TNFalpha. The CIEX method demonstrated that all D2E7 charged-variants bound TNFalpha equally well. There was no preferential binding for any one of D2E7 charged-variants to TNFalpha. D2E7 and 2SD4 Abs were resolved by the CIEX method. When a mixture of D2E7 and 2SD4 was mixed with excess TNFalpha, D2E7. TNFalpha complexes were formed before any 2SD4. TNFalpha complexes. Thus, the CIEX method was able to rank the affinities of the MAbs. D2E7 and TNFalpha formed complexes of 600-5000 kDa. The molecular weights of various D2E7. TNFalpha complexes were determined by a SEC method with light scattering (LS) and refractive index (RI) detectors. Upon overnight incubation, a 598-kDa complex emerged as the most stable and the only D2E7. TNFalpha complex. The molar ratio of D2E7 to TNFalpha in this complex was approximately 1:1. Based on molecular weights and the molar ratio, an immune complex, consisting of alternating three D2E7 and three TNFalpha molecules, is proposed as the most stable complex. PMID- 11730334 TI - Comparison of different protein immobilization methods on quartz crystal microbalance surface in flow injection immunoassay. AB - In this study, a quartz crystal microbalance (QCM) system operated repetitively in flow injection analysis (FIA) mode, is reported. Four immobilization approaches of seven different methods include: (i) physical adsorption; (ii) two thioamine thiolation methods, using cysteamine and cystamine for gold chemisorption and further coupling; (iii) two oxidized dextran spacer methods, coupling of cysteamine and cystamine thiolated QCM surface with periodate oxidized dextran for further Schiff acid-base reaction; and (iv) two thiol-gold chemisorption-based self-assembled monolayer (SAM), applying short-chain, C(3), and long-chain, C(11), mercapto fatty acids to insolubilize human serum albumin (HSA) on QCM surface. Effects of these protein immobilization methods on FIA immunoassay of anti-HSA were compared. At the 0.01 mg/ml anti-HSA level, the lowest analyte concentration tested, the SAM using 11-mercaptoundecanoic acid as QCM surface activating agent generated a larger frequency shift than the other immobilization methods. This implied that the use of thiolated long-chain fatty acid constructed as self-assembled monolayer may thereby potentially be a useful protein immobilization method in QCM-FIA application. PMID- 11730335 TI - Online size-exclusion high-performance liquid chromatography light scattering and differential refractometry methods to determine degree of polymer conjugation to proteins and protein-protein or protein-ligand association states. AB - Characterizing the solution structure of protein-polymer conjugates and protein ligand interactions is important in fields such as biotechnology and biochemistry. Size-exclusion high-performance liquid chromatography with online classical light scattering (LS), refractive index (RI), and UV detection offers a powerful tool in such characterization. Novel methods are presented utilizing LS, RI, and UV signals to rapidly determine the degree of conjugation and the molecular mass of the protein conjugate. Baseline resolution of the chromatographic peaks is not required; peaks need only be sufficiently separated to represent relatively pure fractions. An improved technique for determining the polypeptide-only mass of protein conjugates is also described. These techniques are applied to determining the degree of erythropoietin glycosylation, the degree of polyethylene glycol conjugation to RNase A and brain-derived neurotrophic factor, and the solution association states of these molecules. Calibration methods for the RI, UV, and LS detectors will also be addressed, as well as online methods to determine protein extinction coefficients and dn/dc values both unconjugated and conjugated protein molecules. PMID- 11730336 TI - Peptide mapping and disulfide bond analysis of the cytoplasmic region of an intrinsic membrane protein by mass spectrometry. AB - Intrinsic membrane proteins pose substantial obstacles to analysis by common analytical techniques due to their hydrophobic nature and solubilization requirements. This is the case for studies involving HPLC coupled to mass spectrometry. We have developed an HPLC/mass spectrometry approach to explore and map the peptide sequence of the SERCA1a Ca(2+)-ATPase from the sarcoplasmic reticulum an integral membrane protein of 110 kDa. After extensive proteolysis of the protein, the mass of the proteolytic fragments was analyzed by HPLC/mass spectrometry. Only part of the cytoplasmic fragments was recovered under nondenaturing conditions. On the other hand, peptide fragments obtained under denaturing conditions were found to cover nearly all the cytoplasmic region. Sarcoplasmic reticulum (SR) Ca(2+)-ATPase contains 24 cysteine residues, 18 of which are in the cytosolic or lumenal region of the protein. Peptides containing free cysteines were identified by a mass increase resulting from carboxyamidomethylation of the cysteines with iodoacetamide. Alkylation reactions were executed either before or after reduction of the peptide fragments by dithiothreitol. Analysis of the mass of the fragments indicates that no disulfide bonds exist in the cytoplasmic portion of SR Ca(2+)-ATPase. PMID- 11730337 TI - Determination of carbohydrate contents from excess light scattering. AB - In the light scattering technique, glycosylation gives rise to excess light scattering for glycoproteins. Assuming additivity of refractive index and using an appropriate refractive index increment for carbohydrate, one can determine the degree of glycosylation from the excess light scattering. Here we have used size exclusion chromatography in combination with online light scattering, UV absorbance, and refractive index. The results show that the technique accurately determines the carbohydrate content of recombinant stem cell factor. PMID- 11730338 TI - A nonradioactive high-performance liquid chromatographic microassay for uridine 5'-monophosphate synthase, orotate phosphoribosyltransferase, and orotidine 5' monophosphate decarboxylase. AB - A novel nonradioactive, microassay method has been developed to determine simultaneously the two enzymatic activities of orotate phosphoribosyltransferase (OPRTase) and orotidine 5'-monophosphate decarboxylase (ODCase), either as a bifunctional protein (uridine 5'-monophosphate synthase, UMPS) or as separate enzymes. Substrates (orotate for OPRTase or orotidine 5'-monophosphate for ODCase) and a product (UMP) of the enzymatic assay were separated by high performance liquid chromatography (HPLC) using a reversed-phase column and an ion pairing system; the amount of UMP was quantified by dual-wavelength uv detection at 260 and 278 nm. This HPLC assay can easily detect picomole levels of UMP in enzymatic reactions using low specific activity UMPS of mammalian cell extracts, which is difficult to do with the other nonradioactive assays that have been described. The HPLC assay is suitable for use in protein purification and for kinetic study of these enzymes. PMID- 11730339 TI - Homogeneous time-resolved fluorescence DNA hybridization assay by DNA-mediated formation of an EDTA-Eu(III)-beta-diketonate ternary complex. AB - A sensitive homogenous time-resolved fluorescence DNA hybridization assay method based on the formation of an EDTA-Eu(3+)-beta-diketonate ternary complex in the DNA hybrid was developed. The new approach combined the use of two DNA probes whose sequences compose the whole complementary strand to the target DNA, in which one probe was labeled with an EDTA-Eu(3+) complex on the 5'-terminus and the other, labeled with a bidentate beta-diketone on the 3'-terminus. After hybridization of two DNA probes with target DNA, EDTA-Eu(3+) and beta-diketone come close to each other, and an EDTA-Eu(3+)-beta-diketonate ternary complex with a strong and long-lived fluorescence was formed; thus the target DNA was detected sensitively with a detection limit of 6 pM (0.6 fmol per assay) by time-resolved fluorescence measurement. In the absence of the target DNA, due to the poor stability of bidentate beta-diketonate-Eu(3+) complex in very diluted solution, only a small amount of ternary fluorescence complex was formed. PMID- 11730340 TI - Frontal affinity chromatography coupled to mass spectrometry for screening mixtures of enzyme inhibitors. AB - Frontal affinity chromatography coupled online to mass spectrometry (FAC/MS) has previously been used to estimate binding constants for individual protein ligands present in mixtures of compounds. In this study FAC/MS is used to determine enzyme substrate kinetic parameters and binding constants for enzyme inhibitors. Recombinant human N-acetylglucosaminyltransferase V was biotinylated and adsorbed onto immobilized streptavidin in a microcolumn (20 microL). The enzyme was shown to be catalytically competent transferring GlcNAc from the donor UDP-GlcNAc to beta-d-GlcpNAc-(1-->2)-alpha-d-Manp-(1-->6)-beta-d-Glcp-OR acceptor giving beta-d GlcpNAc-(1-->2)-[beta-d-GlcpNAc-(1-->6)]-alpha-d-Manp-(1-->6)-beta-d-Glcp-OR as the reaction product. The kinetic parameters K(m) and V(max) for the immobilized enzyme could be determined by FAC/MS and were comparable to those measured in solution. Analysis of a mixture of eight trisaccharide analogs in a single run yielded K(d) values for each of the eight compounds ranging from 0.3 to 36 microM. These K(d) values were 2 to 10 times lower than the inhibition constants, K(I)'s, determined in solution using a standard radiochemical assay. However, the ranking order of K(d)'s was the same as the ranking of K(I) values. FAC/MS assays can therefore be employed for the rapid estimation of inhibitor K(d) values making it a valuable tool for enzyme inhibitor evaluations. PMID- 11730341 TI - The advantages of haplotype analysis of the promoter region of the human apolipoprotein E gene. AB - Polymorphisms in the regulatory region of the human apolipoprotein E gene (gene, APOE; protein, apoE) have been implicated in Alzheimer's disease. Here we describe in detail the advantages of a simple method for haplotype analysis of this region (at -491 and -427 bases relative to the transcription start site of the gene). The promoter region of the APOE gene was amplified by polymerase chain reaction (PCR) and this fragment was then used as a template for PCR with "nested" primers to generate a 228-bp product incorporating both the -491 and the -427 loci. PCR products were then digested with DraI and AluI together and subjected to polyacrylamide gel electrophoresis. The distinct pattern of bands appearing on the gel was then used to ascribe [-491,-427] haplotypes to each subject, from which -491 and -427 genotypes were inferred. -491 and -427 genotypes were also confirmed by digestion with DraI alone or AluI alone. Haplotype analysis was successful in all 20 samples analyzed and was 100% consistent with genotyping. We suggest that this is a reliable, time-saving method that the will be useful in large-scale APOE promoter genotyping studies. PMID- 11730343 TI - A fluorescence-based one-step assay for serum non-transferrin-bound iron. AB - We introduce a method for monitoring non-transferrin-bound iron (NTBI), a labile and potentially toxic form of serum iron associated with imbalanced iron metabolism. The assay employs fluorescein-labeled apotransferrin (Fl-aTf), which undergoes fluorescence quenching upon binding iron. It has the advantages of simplicity, high sensitivity, and detection of those forms of NTBI that persist in sera with low transferrin saturations. Since NTBI is not readily available for detection, it is mobilized by 10 mM oxalate. Endogenous serum apotransferrin, capable of binding oxalate-mobilized NTBI, is blocked by 0.1 mM gallium(III). This metal, like iron, binds to Fl-aTf, but it neither quenches its fluorescence nor interferes with quenching by iron. Serum and reagent containing oxalate, Ga(Cl)(3), and Fl-aTf are mixed in multiwell plates and fluorescence is determined after 1 h in a microplate reader. To compensate for artifactual fluorescence changes caused by serum color, parallel samples are prepared with excess unlabeled apotransferrin, which scavenges all iron in the sample. Sera from eight hemochromatosis patients were tested for NTBI by the present assay and by an established alternative method, with qualitatively similar results. A potential application of the test is for screening large numbers of samples from patients at risk of developing NTBI. PMID- 11730342 TI - A quantitative method to measure telomerase activity by bioluminescence connected with telomeric repeat amplification protocol. AB - Telomerase is expected to be a new biomarker for cancer diagnosis. The telomeric repeat amplification protocol (TRAP) is a sensitive method to detect telomerase activity. However, TRAP and its modified protocols are not always suitable for measuring telomerase activity of a large number of clinical samples to diagnosis cancer because these methods generally require a time-consuming detection step such as gel electrophoresis. To improve the procedure for mass diagnosis, we applied bioluminescence to replace the detection step. Telomerase activity is measured by evaluating the amount of inorganic pyrophosphate generated in PCR amplification of telomerase elongation product, with use of the sensitive enzymatic luminometric inorganic pyrophosphate detection assay (ELIDA). TRAP connected with ELIDA (TRAP-ELIDA) can quantitatively detect telomerase activity within linearity from 2 to 1000 cell equivalents. The ELIDA signals accorded with results of TRAP-SYBR green staining, and the results of ELIDA were significantly correlated to those of TRAP connected with an enzyme-linked immunosorbent assay (TRAP-ELISA) (r(2) = 0.992, P < 0.001). TRAP-ELIDA is a simple and sensitive method to quantify telomerase activity without time-consuming gel electrophoresis. Because TRAP-ELIDA measures telomerase activity with a luminometer, it could be applied to a large number of clinical samples at the same time. PMID- 11730344 TI - A new analytical scale DNA affinity binding assay for analyses of specific protein-DNA interactions. AB - We describe a rapid analytical assay for identification of proteins binding to specific DNA sequences. The DAPSTER assay (DNA affinity preincubation specificity test of recognition assay) is a DNA affinity chromatography-based microassay that can discriminate between specific and nonspecific protein-DNA interactions. The assay is sensitive and can detect protein-DNA interactions and larger multicomponent complexes that can be missed by other analytical methods. Here we describe in detail the optimization and utilization of the DAPSTER assay to isolate AP-1 complexes and associated proteins in multimeric complexes bound to the AP-1 DNA element. PMID- 11730345 TI - High-throughput genetic analysis using time-resolved fluorometry and closed-tube detection. AB - Robust methods for genetic analysis are required for efficient exploitation of the constantly accumulating genetic information. We describe a closed-tube genotyping method suitable for high-throughput screening of genetic markers. The method is based on allele-specific probes labeled with an environment-sensitive lanthanide chelate, the fluorescence intensity of which is significantly increased upon PCR amplification of a complementary target. Genomic DNA samples were analyzed in an insulin gene single nucleotide polymorphism (SNP) assay using universal amplification primers and probes that recognized the two different alleles. The feasibility of dry reagent based all-in-one PCR assays was tested using another diabetes-related genetic marker, human leukocyte antigen DQB1 allele *0302 as a model analyte in a dual-color, closed-tube end-point assay. There was a 100% correlation between the novel SNP assay and a conventional PCR restriction fragment length polymorphism assay. It was also demonstrated that using real-time monitoring, accurate genotyping results can be obtained despite strongly cross-reacting probes, minimizing the time and effort needed for optimization of probe sequence. Throughput can be maximized by using predried PCR mixtures that are stable for at least 6 months. This homogenous, all-in-one dry reagent assay chemistry permits cost-effective genetic screening on a large scale. PMID- 11730346 TI - Assay for ADP-ribosyl cyclase by reverse-phase high-performance liquid chromatography. AB - Cyclic ADP-ribose (cADPR), a natural metabolite of beta-NAD(+), is a second messenger for Ca(2+) signaling in T cells. As a tool for purification and identification of ADP-ribosyl cyclase(s) in T cells, a sensitive and specific enzymatic assay using 1,N(6)-etheno-NAD(+) as substrate was developed. A major problem-the sensitivity of 1,N(6)-etheno-cADPR toward the extraction medium perchloric acid-was solved by replacing the perchloric acid extraction procedure of nucleotides by a filtration step. Standard compounds for the HPLC analysis of ADP-ribosyl cyclases and NAD(+)-glycohydrolases, e.g., 1,N(6)-etheno-cADPR, 1,N(6)-etheno-ADPR, and 1,N(6)-etheno-AMP, were produced by ADP-ribosyl cyclase from Aplysia californica and dinucleotide pyrophosphatase. The assay was applied to subcellular fractions prepared from human Jurkat T cells. As a result ADP ribosyl cyclase and NAD(+)-glycohydrolase activity could be detected and precisely quantified in different subcellular fractions indicating the presence of different isoenzymes in T cells. PMID- 11730347 TI - Rapid determination of soluble epoxide hydrolase inhibitors in rat hepatic microsomes by high-performance liquid chromatography with electrospray tandem mass spectrometry. AB - A rapid and reliable electrospray tandem mass spectrometric method for soluble epoxide hydrolase (sEH) inhibitors in rat hepatic microsomes is described. Four synthesized sEH inhibitors were extracted from rat hepatic microsomes with ethyl acetate and were determined by HPLC using positive ion electrospray tandem mass spectrometry within 7 min. The relationship between signal intensity and concentration of sEH inhibitors was linear over the concentration range of 2.0 to 500 ng/mL per 5-microL injection with the use of a noncoeluting internal standard with a similar chemical structure. The intraassay precision was less than 12.4% relative standard deviation and accuracy ranged from -7.0 to 11.3% deviation from the theoretical values with five duplicate assays. The recovery of sEH inhibitors from rat hepatic microsomes, fortified at levels of 50, 100, and 250 ng/mL, averaged 74.2-107.7% with a RSD of 2.1-7.6%. This method was successfully applied to the quantification of residual sEH inhibitors in rat hepatic microsomes without interference. PMID- 11730348 TI - A potential role for the analytical ultracentrifuge in the experimental measurement of protein valence. AB - A method is described whereby sedimentation velocity is combined with equilibrium dialysis to determine the net charge (valence) of a protein by using chromate as an indicator ion for assessing the extent of the Donnan redistribution of small ions. The procedure has been used in experiments on bovine serum albumin under slightly alkaline conditions (pH 8.0, I 0.05) to illustrate its application to a system in which the indicator ion and protein both bear net negative charge and on lysozyme under slightly acidic conditions (pH 5.0, I 0.10) to illustrate the situation where chromate is a counterion. PMID- 11730349 TI - Determination of monoenzyme- and bienzyme-stimulated precipitation by a cuvette based surface plasmon resonance instrument. AB - This paper describes the use of a cuvette-based surface plasmon resonance (SPR) instrument to measure biocatalyzed precipitation reactions. Enzyme-modified SPR sensor disk forms the base of a cuvette, in which the substrate solution is added with stirring. The determination of the substrate concentration relies on the measurement of SPR angle shift (Deltatheta(SPR)) induced by the deposition of the insoluble products without involving in any electrochemical reactions. As examples, horseradish peroxidase (HRP)-modified monoenzyme SPR sensor and HRP glucose oxidase bienzyme-layered sensor are created to determine hydrogen peroxide and glucose via the catalyzed oxidation of 4-chloro-1-naphthol (4-CN). The deposition of the oxidized 4-CN-insoluble products leads to SPR angle shifts, which are linear to H(2)O(2) and glucose in the concentration ranges of 0.067 7.24 x 10(-5) and 0.7-8.3 x 10(-4) mM, respectively. The SPR sensitivities are greater than those of nonelectrochemical quartz crystal microbalance (QCM) (the parallel results in this study) and compare favorable with those of electrochemical QCM and electrochemical SPR methods. This study opens the field for enhanced SPR measurements by using biocatalyzed precipitation as a signal amplification method. PMID- 11730350 TI - 5,6-carboxyfluorescein diacetate succinimidyl ester-labeled apoptotic and necrotic as well as detergent-treated cells can be traced in composite cell samples. AB - Detection of dividing cells by staining with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) has been widely used in flow-cytometric protocols. We analyzed the fate of CFSE in cells undergoing apoptotic or necrotic cell death, respectively. Peripheral blood mononuclear cells (PBMC) were stained with CFSE. Apoptosis was induced by UVB irradiation and necrosis by incubation at 56 degrees C for 30 min. In some experiments, labeled cells were permeabilized with detergent and CFSE association with nuclei was assessed. We observed that (i) CFSE remains stably detectable in apoptotic and necrotic cells; (ii) CFSE remains stably associated with the nuclei of cells even after their lysis by detergent; (iii) CFSE labeling does not interfere with the induction of cell death; and (iv) CFSE is not transferred from stained dying cells to unstained neighboring counterparts. We conclude that, in addition to tracking viable cells, CFSE can be used to trace dying cells in composite samples. We demonstrated that CFSE labeling does not influence the induction and the execution of apoptosis or necrosis. PMID- 11730351 TI - Assays of methylenetetrahydrofolate reductase and methionine synthase activities by monitoring 5-methyltetrahydrofolate and tetrahydrofolate using high performance liquid chromatography with fluorescence detection. AB - We developed a method for assays of methylenetetrahydrofolate reductase and methionine synthase activities by monitoring their products of 5 methyltetrahydrofolate (5-CH(3)-H(4)folate) and tetrahydrofolate (H(4)folate) directly, using high-performance liquid chromatography with fluorescence detection. Folate derivatives and enzymes were stable in the assay process. No reagents in the assay mixture were found to disturb the separation and detection of both H(4)folate and 5-CH(3)-H(4)folate in our assay system. The detection limit of this method was less than 20 nM H(4)folate or 5-CH(3)-H(4)folate in the enzyme assay system. This analytical method, therefore, has a sensitivity high enough to obtain accurate parameters of Michaelis-Menten kinetics and for assays of crude extracts from various biological samples. In addition, the analytical procedure is very simple and economical; it may be a useful tool for studying methylenetetrahydrofolate reductase and methionine synthase activities. PMID- 11730352 TI - In-gel digestion of proteins from long-term dried polyacrylamide gels: matrix assisted laser desorption-ionization time of flight mass spectrometry identification of proteins and detection of their covalent modification. PMID- 11730353 TI - Using magnetic dipole transitions for fluorescence resonance energy transfer. PMID- 11730354 TI - Determination of Ca(2+) influx across plant plasma membrane with sealed vesicles reoriented by Brij 58. PMID- 11730355 TI - Calsequestrin blot overlay of two-dimensional electrophoretically separated microsomal proteins from skeletal muscle. PMID- 11730356 TI - Analysis of protein circular dichroism spectra based on the tertiary structure classification. PMID- 11730357 TI - Detection of antibacterial polypeptide activity in situ after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 11730358 TI - Standardization of nitric oxide aqueous solutions by modified Saltzman method. AB - Nitric oxide (NO) aqueous solutions were prepared by saturating pure NO gas and hydrolyzing 1 mM 1-hydroxy-2-oxo-3-(N-methyl-3-aminoethyl)-3-methyl-1-triazene (NOC-7), a NO donor, under anerobic conditions. The modified Saltzman method was employed for standardization of the NO aqueous solutions. NO and NO(2) in the solutions were driven with nitrogen gas stream into the first Saltzman solution to measure NO(2) and the leaked NO was driven with air stream through an oxidizing solution into the second Saltzman solution to measure NO, and NO(-)(2) and NO(-)(3) in the residual solutions were determined directly and after reduction with nitrate reductase, respectively. The concentrations of nitrogen oxide species in the NO solutions were about 1.8 mM NO/0.01 mM NO(2)/0.1 mM NO( )(2)/0.1 mM NO(-)(3), and unchanged during keeping at 20 degrees C for 1 h under anerobic conditions but became 0.05 mM NO/0.01 mM NO(2)/1.7 mM NO(-)(2)/0.1 mM NO(-)(3) by keeping at 20 degrees C for 10 min under aerobic conditions. Instability of NO under aerobic conditions was supported by consumption of 1/4 equivalent amount of dissolved oxygen, and by loss of ability to convert 2-(4 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO) to carboxy-PTI. Simultaneous quantification of nitrogen oxide species by the modified Saltzman method was found to be useful for practical standardization of NO aqueous solutions. PMID- 11730359 TI - Nitric oxide inhibition enhances platelet aggregation in experimental anti-Thy-1 nephritis. AB - In the present paper we studied the role of nitric oxide radicals (NO) on platelet aggregation, fibrinogen deposition, superoxide formation, peroxynitrite formation, hemodynamics, and leukocyte migration in the Thy-1 model of glomerulonephritis. To first study the baseline kinetics of these parameters, groups of anti-Thy-1-treated rats were sacrificed at 1 h, 4 h, 24 h, 3 days, 7 days, and 14 days and compared to controls. Urinary protein excretion was significantly elevated in Thy-1 nephritis at 3 and 7 days. Glomerular macrophages, PMNs, and superoxide anion-positive cells were significantly increased in Thy-1 nephritis. Nitrotyrosine immunoreactivity was absent during the entire study period. Glomerular platelet aggregation was significantly increased in anti-Thy-1 injected rats at 1 h, 4 h, 24 h, and 3 days. Glomerular fibrinogen deposition was significantly elevated at all time points. To elucidate the role of NO in this process, additional groups of anti-Thy-1-injected rats were treated with the NOS inhibitor l-NAME and studied at 24 h. Urinary protein excretion was significantly higher in l-NAME treated Thy-1 rats compared to nontreated Thy-1 rats. Plasma and urine nitrite/nitrate levels were significantly lower in l-NAME-treated Thy-1 rats compared to nontreated Thy-1 rats. Compared to nontreated Thy-1 rats, there were no differences in intraglomerular leukocyte accumulation after treatment with l-NAME. In contrast, we observed a marked increase in platelet aggregation following l-NAME treatment. From these data we conclude that the inflammatory infiltrate in Thy-1 nephritis develops independent of NO radical production, whereas NO radicals prevent the accumulation of platelet aggregates. PMID- 11730360 TI - Mechanisms of synergistic cytokine-induced nitric oxide production in human alveolar epithelial cells. AB - Nitric oxide (NO) derived from inducible NO synthase (iNOS) at sites of inflammation is closely related to host defense against infection and airway inflammation. Cytokines are known to stimulate NO production in human alveolar epithelial cells in a synergistic (nonlinear or nonadditive) manner. The mechanism of this synergy is not known. We measured the activation of the transcription factor NF-kappaB, the iNOS protein, and NO production in A549 monolayers (human alveolar epithelial cell line) in response to different combinations of IL-1beta, INF-gamma, and TNF-alpha (100 ng/ml), and the cofactors FMN, FAD, and BH4. We found that both IL-1beta and TNF-alpha could independently activate cytosolic NF-kappaB, direct its translocation into the nucleus, and induce iNOS monomer synthesis. In addition, different combinations of cytokines produced synergistic amounts of iNOS monomers. Exogenous BH4 (0.1 microM) had no impact on NO production induced by cytokine combinations that included IL-1beta, but significantly enhanced NO production in the presence of INF-gamma and TNF alpha, and allowed TNF-alpha independently to produce NO. We conclude that there are at least three mechanisms of synergistic cytokine-induced NO production: (1) the biosynthesis of iNOS monomer due to nonlinear interactions by transcription factors, (2) synergistic cytosolic activation of NF-kappaB, and (3) parallel biosynthesis of BH4 in the presence of cytokine combinations that include IL 1beta. PMID- 11730361 TI - Lipopolysaccharide temporarily impairs sphincter of Oddi motility. AB - The aim of our study was to determine the effect of lipopolysaccharide (LPS) on sphincter of Oddi (SO) motility. Opossums received saline, Escherichia coli LPS (1.0 mg/kg), or E. coli LPS (1.0 mg/kg) and aminoguanidine (50 mg/kg), and the SO was removed 6-24 h later. At 12 h LPS decreased electrical field stimulation (EFS)-induced relaxation and increased baseline tone. These changes were reversed when the animals were pretreated with aminoguanidine. The dose-dependent decrease in EFS-induced relaxation by N(omega)-nitro-l-arginine was impaired after LPS, but not in animals that received LPS and aminoguanidine. The impaired EFS-induced relaxation after LPS was reversed when l-arginine was added to the tissue bath. Serum levels of NO(-)(2)/NO(-)(3) were increased with LPS as compared to saline or both LPS and aminoguanidine. Inducible nitric oxide synthase mRNA was readily seen in SO segments after LPS. LPS impairs EFS-induced relaxation and increases baseline tone of the SO. The effects of LPS on SO motility appear to be mediated by nitric oxide. PMID- 11730362 TI - Protection of human keratinocyte mtDNA by low-level nitric oxide. AB - This study was designed to evaluate the DNA damaging effects of nitric oxide and to determine whether the endogenous generation of nitric oxide at low levels in the cell exerts a protective effect against this damage. Damage to mitochondrial and nuclear DNA in normal human epidermal keratinocytes (NHEK) was assessed after treatment of these cells with varying concentrations of S-nitroso-N acetylpenicillamine, which decomposes to release nitric oxide. The results showed that mitochondrial DNA was more vulnerable to nitric oxide-induced damage than was a similarly sized fragment of the beta-globin gene. To evaluate the effects on DNA damage by pretreatment of cells with low-levels of nitric oxide, NHEK cells were treated with the prodrug V-PYRRO/NO. This agent is metabolized inside these cells and releases small quantities of nitric oxide. The cells then were exposed to damaging amounts of nitric oxide produced by S-nitroso-N acetylpenicillamine. The results of these studies showed that pretreatment of NHEK cells with V-PYRRO/NO attenuated the mtDNA damage and loss of cell viability produced by exposure to S-nitroso-N-acetylpenicillamine. PMID- 11730363 TI - In situ measurement of nitric oxide production in cardiac isografts and rejecting allografts by an electrochemical method. AB - A number of previous studies have indirectly (electron paramagnetic resonance, nitrite/nitrate, ribonuclease protection assay for inducible nitric oxide synthase (iNOS) mRNA, l-citrulline assay) demonstrated the production of nitrogen monoxide (NO) during early cardiac allograft rejection. This study reports the first direct, quantitative measurement using an electrochemical method of NO produced from rejecting allograft tissue studied in vitro. A rat heterotopic abdominal transplant preparation was utilized. Day 7 isograft (ACI to ACI) or allograft (Lewis to ACI) transplanted hearts were atraumatically harvested and suspended at 4 degrees C in Ringers-Hepes solution. An electrochemical system highly sensitive and specific for NO consisting of a Nafion-coated platinum disk electrode (lower limit, 50 nM NO) coupled to an analysis system measured ongoing oxidation of NO. Measurements were carried out after inserting the electrode in the tissue block and warming the block to 25 degrees C. Additional measurements were also made after incubation of tissue with aminoguanidine (AG), a relatively selective iNOS inhibitor. Direct measurements (mean +/- SEM) from allograft tissue indicated a fourfold increase in NO as compared with isografts (13.41 +/- 4.40 microM NO vs. 3.43 +/- 2.04 microM NO). Incubation of allograft tissue with AG reduced NO levels to isograft levels (13.41 +/- 4.40 microM NO vs. 5.94 +/- 3.14 microM NO); AG had no effect on measured isograft NO levels. Direct, quantitative measurement of NO from tissue is feasible and reproducible, and discrimination between different levels of NO production can be made. These results confirm the imputed results from the previous studies using this experimental model. This technology promises to be a valuable tool for evaluating specific modulators of NO production studied under a variety of physiologic and pathophysiologic conditions. PMID- 11730365 TI - Effects of pH, nitrite, and ascorbic acid on nonenzymatic nitric oxide generation and bacterial growth in urine. AB - Nitrite may be generated by bacteria in urine during urinary tract infections. Acidification of nitrite results in the formation of nitric oxide (NO) and other reactive nitrogen oxides, which are toxic to a variety of microorganisms. We have studied NO formation and bacterial growth in mildly acidified human urine containing nitrite and the reducing agent vitamin C. Urine collected from healthy subjects was incubated in closed syringes at different pH values with varying amounts of nitrite and/or ascorbic acid added. NO generation was measured in headspace gas using a chemiluminescence technique. A similar setup was also used to study the growth of three strains of bacteria in urine. Mildly acidified nitrite-containing urine generated large amounts of NO and this production was greatly potentiated by ascorbic acid. The growth of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus saprophyticus was markedly reduced by the addition of nitrite to acidified urine. This inhibition was enhanced by ascorbic acid. In conclusion, we show that the growth of three common urinary pathogens is markedly inhibited in mildly acidified urine when nitrite is present. The bacteriostatic effect of acidified nitrite is likely related to the release of NO and other toxic reactive nitrogen intermediates. These results may help to explain the well known beneficial effects of urinary acidification with, e.g., vitamin C in treatment and prevention of urinary tract infection. PMID- 11730364 TI - Regulation of nitric oxide production by murine peritoneal macrophages treated in vitro with chemokine monocyte chemoattractant protein 1. AB - Monocyte chemoattractant protein 1 (MCP-1) is an important mediator of monocyte/macrophage recruitment and activation at the sites of chronic inflammation and neoplasia. In the current study, the role of nitrogen monoxide (NO) in the activation of murine peritoneal macrophages to the tumoricidal state in response to in vitro MCP-1 treatment and the regulatory mechanisms involved therein were investigated. Murine peritoneal macrophages upon activation with MCP 1 showed a dose- and time-dependent production of NO together with increased tumoricidal activity against P815 mastocytoma cells. N-monomethyl-l-arginine (L NMMA), a specific inhibitor of the l-arginine pathway, inhibited the MCP-1 induced NO secretion and generation of macrophage-mediated tumoricidal activity against P815 (NO-sensitive, TNF-resistant) cells but not the L929 (TNF-sensitive, NO-resistant) cells. These results indicated l-arginine-dependent production of NO to be one of the effector mechanisms contributing to the tumoricidal activity of MCP-1-treated macrophages. Supporting this fact, expression of iNOS mRNA was also detected in the murine peritoneal macrophages upon treatment with MCP-1. Investigating the signal transduction pathway responsible for the NO production by the MCP-1-activated murine peritoneal macrophages, it was observed that the pharmacological inhibitors wortmannin, H-7 (1-(5-isoquinoline sulfonyl)-2-methyl piperazine dihydrochloride), and PD98059 blocked the MCP-1-induced NO production, suggesting the probable involvement of phosphoinositol-3-kinase, protein kinase C, and p42/44 MAPkinases in the above process. Various modulators of calcium and calmodulin (CaM) such as EGTA, nifedipine, TMB-8 (3,4,5-trimethoxybenzoic acid-8 (diethylamino)octyl ester), A23187, and W-7 (N-(6-aminohexyl)-5-chloro-1 napthalenesulfonamide) were also found to modulate the in vitro macrophage NO release in response to MCP-1. This observation indicated the regulatory role of calcium/CaM in the process of MCP-1-induced macrophage NO production. Similarly, the role of serine/threonine and protein tyrosine phosphatases in the above pathway was suggested using the specific inhibitors of these phosphatases, okadaic acid and sodium orthovanadate. PMID- 11730366 TI - Systemic biochemical effects of inhaled NO in rats: increased expressions of NOS III, nitrotyrosine-, and phosphotyrosine-immunoreactive proteins in liver and kidney tissues. AB - Inhaled nitric oxide (iNO) has been shown to reduce pulmonary hypertension associated with several disease states. The effects of iNO are thought to be restricted to the pulmonary vasculature because of its rapid inactivation by hemoglobin. Recent data have suggested, however, that iNO can form nitrosothiols, which can be carried throughout the circulation, thus increasing the half life and bioactivity on NO. Other studies have shown that iNO can affect intestinal ischemia and renal hemodynamics. In this study, rats were exposed to 49 +/- 4 ppm or 107 +/- 13 ppm NO for 4 h and the lung, spleen, liver, and kidney tissues were removed and measured for NOS II and NOS III protein, nitrotyrosine (NT), and phosphotyrosine (PT) immunoreactivity. Following 107 ppm iNO, increases in NOS III protein expression, NT, and PT were observed in the liver and kidney, but not in the lung or spleen. No such increases were noted after the lower dose of iNO. These results paralleled those shown for isobutyl nitrite that we reported earlier and indicated that iNO can cause changes in protein chemistry in organs and tissues beyond the lungs. Since iNO produced little systemic hemodynamic effects, it is unlikely that the observed biochemical alterations were derived secondarily from physiological changes. PMID- 11730394 TI - Estrogen in the prevention of atherosclerosis. A randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Although observational studies suggest that estrogen replacement therapy (ERT) reduces cardiovascular morbidity and mortality in postmenopausal women, use of unopposed ERT for prevention of coronary heart disease in healthy postmenopausal women remains untested. OBJECTIVE: To determine the effects of unopposed ERT on the progression of subclinical atherosclerosis in healthy postmenopausal women without preexisting cardiovascular disease. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: University-based clinic. PATIENTS: 222 postmenopausal women 45 years of age or older without preexisting cardiovascular disease and with low-density lipoprotein cholesterol levels of 3.37 mmol/L or greater (>/=130 mg/dL). INTERVENTION: Unopposed micronized 17beta-estradiol (1 mg/d) or placebo. All women received dietary counseling. Women received lipid-lowering medication if their low-density lipoprotein cholesterol level exceeded 4.15 mmol/L (160 mg/dL). MEASUREMENTS: The rate of change in intima-media thickness of the right distal common carotid artery far wall in computer image processed B-mode ultrasonograms obtained at baseline and every 6 months during the 2-year trial. RESULTS: In a multivariable mixed-effects model, among women who had at least one follow-up measurement of carotid intima-media thickness (n = 199), the average rate of progression of subclinical atherosclerosis was lower in those taking unopposed estradiol than in those taking placebo (-0.0017 mm/y vs. 0.0036 mm/y); the placebo-estradiol difference between average progression rates was 0.0053 mm/y (95% CI, 0.0001 to 0.0105 mm/y) (P = 0.046). Among women who did not receive lipid-lowering medication (n = 77), the placebo-estradiol difference between average rates of progression was 0.0147 mm/y (CI, 0.0055 to 0.0240) (P = 0.002). Average rates of progression did not differ between estradiol and placebo recipients who took lipid-lowering medication (n = 122) (P > 0.2). CONCLUSIONS: Overall, the average rate of progression of subclinical atherosclerosis was slower in healthy postmenopausal women taking unopposed ERT with 17beta-estradiol than in women taking placebo. Reduction in the progression of subclinical atherosclerosis was seen in women who did not take lipid-lowering medication but not in those who took these medications. PMID- 11730396 TI - Predictors of virologic and clinical outcomes in HIV-1-infected patients receiving concurrent treatment with indinavir, zidovudine, and lamivudine. AIDS Clinical Trials Group Protocol 320. AB - BACKGROUND: A substantial proportion of patients with HIV infection will not respond to antiretroviral therapy. Early predictors of response to treatment are needed to identify patients who are at risk for treatment failure. OBJECTIVE: To determine predictors of virologic and clinical response to indinavir, zidovudine, and lamivudine therapy. DESIGN: Observational analysis of one treatment group in a phase III trial. SETTING: 40 AIDS Clinical Trials units. PATIENTS: 489 patients receiving indinavir, zidovudine, and lamivudine who had 1) a CD4 count of 0.200 x 10(9) cells/L or less after 8 or more weeks of study therapy and 2) plasma HIV-1 RNA measurements obtained at baseline and week 8. MEASUREMENTS: HIV-1 RNA level and CD4 cell count at weeks 0, 4, 8, 24, and 40. Clinical progression was defined as a new AIDS-defining illness or death. RESULTS: Patients' levels of HIV-1 RNA at the 8th study week of therapy predicted whether patients would achieve virologic suppression to below 500 (or 50) copies/mL at study week 24. An HIV-1 RNA level less than 500 copies/mL at week 24 was achieved in 71% of patients whose level at week 8 had been less than 500 copies/mL, 53% of those with a level of 500 copies/mL or more and at least 2-log(10) copies/mL reduction since baseline, 29% of those with a level of 500 copies/mL or more with a 1- to 1.99 log(10) copies/mL reduction, and 9% of those with a level of 500 copies/mL or greater and less than 1-log(10) copies/mL reduction since baseline (P < 0.001). HIV-1 RNA level at week 8 also predicted clinical progression. HIV-1 disease progressed in 2.2% of the patients with a week-8 HIV-1 RNA level less than 500 copies/mL, 2.3% of patients with 500 copies/mL or greater and at least 2-log(10) copies/mL reduction since baseline, 4.9% of patients with 500 copies/mL or greater and 1- to 1.99-log(10) copies/mL reduction since baseline, and 10.6% of patients with 500 copies/mL or greater and less than 1-log(10) copies/mL decrease since baseline (P = 0.009). After adjustment for HIV-1 RNA level, patients with a higher week-8 CD4 cell count were more likely to have a week-24 HIV-1 RNA level less than 500 copies/mL (relative risk for patients with a week-8 CD4 count >/= 0.10 x 10(9) cells/L, 1.47 [95% CI, 1.00 to 2.16] compared with <0.050 x 10(9) cells/L; relative risk for patients with a week-8 CD4 count of 0.05 to 0.099 x 10(9) cells/L, 0.98 [CI, 0.61 to 1.57] compared with <0.050 x 10(9) cells/L). After adjustment for HIV-1 RNA level, patients with a week-8 CD4 count of 0.05 x 10(9) cells/L or greater (compared with <0.05 x 10(9) cells/L) had a decreased hazard for clinical progression (hazard ratio, 0.25 [CI, 0.09 to 0.67]). CONCLUSIONS: The HIV-1 RNA level and CD4 cell count achieved at 8 weeks of treatment are important predictors of subsequent virologic and clinical outcomes. PMID- 11730397 TI - Resistance training to counteract the catabolism of a low-protein diet in patients with chronic renal insufficiency. A randomized, controlled trial. AB - BACKGROUND: Chronic renal insufficiency leads to muscle wasting, which may be exacerbated by low-protein diets prescribed to delay disease progression. Resistance training increases protein utilization and muscle mass. OBJECTIVE: To determine the efficacy of resistance training in improving protein utilization and muscle mass in patients with chronic renal insufficiency treated with a low protein diet. DESIGN: Randomized, controlled trial. SETTING: Tufts University, Boston, Massachusetts. PATIENTS: 26 older patients with moderate renal insufficiency (17 men, 9 women) who had achieved stabilization on a low-protein diet. INTERVENTION: During a run-in period of 2 to 8 weeks, patients were instructed and their adherence to the low-protein diet (0.6 g/kg of body weight per day) was evaluated. They were randomly assigned to a low-protein diet plus resistance training (n = 14) or a low-protein diet alone (n = 12) for 12 weeks. MEASUREMENTS: Total body potassium, mid-thigh muscle area, type I and II muscle fiber cross-sectional area, and protein turnover. RESULTS: Mean protein intake was 0.64 +/- 0.07 g/kg per day after stabilization. Total body potassium and type I and II muscle-fiber cross-sectional areas increased in patients who performed resistance training by a mean (+/-SD) of 4% +/- 8%, 24% +/- 31%, and 22% +/- 29%, respectively, compared with those who did not. Leucine oxidation and serum prealbumin levels also improved significantly. Patients assigned to resistance training maintained body weight compared with those who were not. Improvement in muscle strength was significantly greater with resistance training (32% +/- 14%) than without (-13% +/- 20%) (P < 0.001). CONCLUSION: By improving muscle mass, nutritional status, and function, resistance training seems to be effective against the catabolism of a low-protein diet and uremia in patients with renal failure. PMID- 11730398 TI - Menstrual cyclic variation of myocardial ischemia in premenopausal women with variant angina. AB - BACKGROUND: An abundance of ovarian hormones is assumed to be a major contributor to the low incidence of ischemic heart disease in premenopausal women. However, the effects of ovarian hormones remain undetermined. OBJECTIVE: To examine whether the variation in ovarian hormone levels throughout a menstrual cycle affects myocardial ischemia in women with variant angina. DESIGN: Prospective, observational study. SETTING: University medical center in Japan. PARTICIPANTS: 10 premenopausal women with variant angina. MEASUREMENTS: Frequency of spontaneous ischemic episodes, flow-mediated dilation of brachial artery, and serum levels of estradiol and progesterone. RESULTS: Frequency of ischemic episodes was highest from the end of the luteal phase to the beginning of the menstrual phase and was lowest in the follicular phase. Flow-mediated vasodilation and estradiol levels were lowest from the end of the luteal phase to the beginning of the menstrual phase and were highest in the follicular phase. CONCLUSIONS: In premenopausal women with variant angina, we documented a cyclic variation in endothelial function and the frequency of myocardial ischemia that was associated with the variation in estrogen levels. PMID- 11730399 TI - Reported methodologic quality and discrepancies between large and small randomized trials in meta-analyses. AB - PURPOSE: To explore whether reported methodologic quality affects estimated intervention effects in randomized trials and contributes to discrepancies between the results of large randomized trials and small randomized trials in meta-analyses. DATA SOURCES: Meta-analyses of randomized trials that included at least one large trial (>/=1000 participants) were included, regardless of the therapeutic area. Eligible meta-analyses were identified through electronic searches and bibliographies of relevant articles. STUDY SELECTION: Full-length randomized trials. DATA EXTRACTION: Methodologic quality was assessed according to reported randomization, double blinding, and follow-up as separate components and by using the Jadad composite scale. DATA SYNTHESIS: Fourteen meta-analyses involving 190 randomized trials from eight therapeutic areas were included. Compared with large trials, intervention effects were exaggerated in small trials with inadequate allocation sequence generation (ratio of odds ratios, 0.46 [95% CI, 0.25 to 0.83]; P = 0.011), inadequate allocation concealment (ratio of odds ratios, 0.49 [CI, 0.27 to 0.86]; P = 0.014), and no double blinding (ratio of odds ratios, 0.52 [CI, 0.28 to 0.96]; P = 0.01). Large trials did not differ significantly from small trials with adequate generation of the allocation sequence, adequate allocation concealment, or adequate double blinding. No association was seen between reported follow-up and intervention effects. The Jadad scale provided no additional information because the scale and the quality components overlapped substantially. CONCLUSIONS: Inadequate generation of the allocation sequence, allocation concealment, and double blinding lead to exaggerated estimates of intervention benefit and may contribute to discrepancies between the results of large randomized trials and small randomized trials in meta-analyses. PMID- 11730400 TI - Automated external defibrillators: technical considerations and clinical promise. AB - Early defibrillation is the most important determinant of survival for victims of cardiac arrest due to ventricular fibrillation. The automated external defibrillator (AED) was developed as the result of the American Heart Association's Public Access Defibrillation initiative. The goal of this initiative is to place AEDs in strategic locations so that laypersons with minimal training could promptly defibrillate victims of cardiac arrest. Because of changes in design and the use of alternative waveforms for defibrillation, the modern AED is compact and portable, simple to use, and highly efficacious; in addition, it requires little maintenance. Automated external defibrillators have been used successfully by traditional and nontraditional responders as well as laypersons. In special environments, such as casinos and commercial aircraft, AEDs have performed particularly well. State and federal legislation has eased concerns about AED use by extending legal protection to AED users under Good Samaritan laws. Since the experience continues to be positive, AEDs are being used in increasingly diverse community locations, and public awareness is growing. The American Heart Association's initiative is progressing rapidly. PMID- 11730401 TI - Resistance training enhances the value of protein restriction in the treatment of chronic kidney disease. PMID- 11730402 TI - Coronary artery disease in young women: the menstrual cycle and other risk factors. PMID- 11730403 TI - Clinical trial acronyms. PMID- 11730404 TI - Clinical trial acronyms. PMID- 11730406 TI - Methotrexate in giant-cell arteritis. PMID- 11730407 TI - Methotrexate in giant-cell arteritis. PMID- 11730409 TI - Nonhypoglycemic effects of thiazolidinediones. PMID- 11730410 TI - New psychopharmacologic treatment strategies. PMID- 11730412 TI - Albumin meta-analysis. PMID- 11730414 TI - Clinical librarianship. PMID- 11730415 TI - Sympathetic neurocirculatory failure in Parkinson disease: Evidence for an etiologic role of alpha-synuclein. PMID- 11730416 TI - Severe neurotoxicity of cefepime in uremic patients. PMID- 11730421 TI - Public health triumphs at WTO Conference. PMID- 11730422 TI - Anesthesiologists urge safe, ethical practice. PMID- 11730423 TI - From the Food and Drug Administration. PMID- 11730426 TI - Academic alert. Midwest trials of heart-assist device. PMID- 11730427 TI - Brain injury resulting from falls among elderly persons. PMID- 11730429 TI - Graduating residents' perceptions of their preparedness for practice. PMID- 11730430 TI - Graduating residents' perceptions of their preparedness for practice. PMID- 11730431 TI - Graduating residents' perceptions of their preparedness for practice. PMID- 11730433 TI - Physical activity counseling in primary care. PMID- 11730434 TI - Meningococcal disease in college students. PMID- 11730436 TI - Using meta-analysis to answer clinical questions. PMID- 11730437 TI - Using meta-analysis to answer clinical questions. PMID- 11730441 TI - Toxic landscaping of facilities for developmentally disabled adults. PMID- 11730442 TI - Aptiganel hydrochloride in acute ischemic stroke: a randomized controlled trial. AB - CONTEXT: Tissue plasminogen activator is the only thrombolytic agent approved in the United States for treatment of acute ischemic stroke, and has limitations. Aptiganel hydrochloride is a novel and selective ligand for the ion-channel site of the N-methyl-D-aspartate receptor-channel complex and a promising neuroprotective agent in animal models of focal brain ischemia. OBJECTIVE: To determine whether aptiganel improves the clinical outcome for acute ischemic stroke patients. DESIGN: Nested phase 2/phase 3 randomized controlled trial conducted between July 1996 and September 1997. SETTING: One hundred fifty-six medical centers in the United States, Canada, Australia, South Africa, England, and Scotland. PARTICIPANTS: A total of 628 patients with hemispheric ischemic stroke (50.3% male; mean age, 71.5 years). INTERVENTIONS: Patients were randomly assigned within 6 hours of stroke to receive 1 of 3 treatment regimens: high-dose aptiganel (5-mg bolus followed by 0.75 mg/h for 12 hours; n = 214); low-dose aptiganel (3-mg bolus followed by 0.5 mg/h for 12 hours; n = 200); or placebo (n = 214). MAIN OUTCOME MEASURES: The primary efficacy end point was the Modified Rankin Scale score at 90 days after stroke onset. Secondary end points included mortality and change in National Institutes of Health (NIH) Stroke Scale score at 7 days after stroke. RESULTS: The trial was suspended by the sponsor and the independent data and safety monitoring board because of both a lack of efficacy and a potential imbalance in mortality. There was no improvement in outcome for either aptiganel (low-dose or high-dose) group compared with the placebo group at 90 days (median Modified Rankin Scale score for all 3 treatment groups = 3; P =.31). At 7 days, placebo-treated patients exhibited slightly greater neurological improvement on the NIH Stroke Scale than high-dose aptiganel patients (mean improvement for placebo group, -0.8 points vs for high-dose aptiganel, 0.9 points; P =.04). The mortality rate at 120 days in patients treated with high-dose aptiganel was higher than that in patients who received placebo (26.3% vs 19.2%; P =.06). Mortality in the low-dose aptiganel group was 22.5% (P =.39 vs placebo). CONCLUSIONS: Aptiganel was not efficacious in patients with acute ischemic stroke at either of the tested doses, and m ay be harmful. The larger proportion of patients with favorable outcomes and lower mortality rate in the placebo group suggest that glutamate blockade with aptiganel may have detrimental effects in an undifferentiated population of stroke patients. PMID- 11730443 TI - Survival effect of lung transplantation among patients with cystic fibrosis. AB - CONTEXT: Patients with cystic fibrosis (CF) are the second largest group of lung transplant recipients in the United States. The survival effect of transplantation on a general CF population has not previously been measured. OBJECTIVE: To determine the impact of bilateral lung transplantation on survival in patients with CF. DESIGN, SETTING, AND PATIENTS: Retrospective observational cohort study of 11 630 CF patients who did not undergo lung transplantation (controls) and 468 transplant recipients with CF from 115 CF centers in the United States, 1992-1998. Patients were stratified into 5 groups based on a 5 year survival prediction model (survival group 1: <30%; survival group 2: 30 to <50%; survival groups 3-5: 50 to <100%.) MAIN OUTCOME MEASURE: Five-year survival from date of transplantation in 1992-1997 in the transplant group and from January 1, 1993, in the control group. RESULTS: Lung transplantation increased 5 year survival of CF patients in survival group 1. Survival group 2 had equivocal survival effects, and groups 3-5 had negative survival effects from transplantation. From 1994-1997, there was a mean annual prevalence of 238 patients in survival group 1 and mean annual incidence of 154 patients entering the group, approximately 1.5 times the number of lung transplantations performed each year in CF patients (mean, 104). Use of the criterion of forced expiratory volume in 1 second of less than 30% resulted in an equivocal survival benefit and identified 1458 potential candidates for transplantation in 1993. CONCLUSIONS: Cystic fibrosis patients in group 1 have improved 5-year survival after lung transplantation. The majority of patients with CF have equivocal or negative survival effects from the procedure. Selection of patients with CF for transplantation based on group 1 survival predictions maximizes survival benefits to individuals and may reduce the demand for scarce donor organs. PMID- 11730444 TI - Survival by time of day of hemodialysis in an elderly cohort. AB - CONTEXT: Patients with end-stage renal disease (ESRD) typically undergo hemodialysis (HD) during the morning or afternoon, with time of treatment generally based on space availability or patient preference. No studies have investigated variation in patient survival as a function of the time of day when they receive dialysis. OBJECTIVE: To investigate the association of elderly patients' HD treatment shift with their continued survival, controlling for well established HD-related mortality risk factors. DESIGN, SETTING, AND PARTICIPANTS: An 11-year follow-up cohort study conducted among 242 ESRD patients aged 60 years or older who underwent HD at 58 dialysis facilities in Georgia either during a morning shift (n = 167) or an afternoon shift (n = 75) and who completed baseline (1988) [corrected] and follow-up (1991) interviews. MAIN OUTCOME MEASURE: Mortality from all causes occurring through July 7, 1999, as verified by death certificate reviews, and compared by morning vs afternoon-shift HD. RESULTS: Morning-shift HD patients survived significantly longer than afternoon-shift patients (median survival, 941 days vs 470 days; P<.001). A Cox proportional hazards model indicated that the morning shift was protective (relative risk, 0.71; 95% confidence interval, 0.53-0.95) independent of age, race, sex, body mass index, functional status, diabetic ESRD, cardiovascular comorbidity, weekly hours of dialysis, and months of dialysis. CONCLUSIONS: Possible explanations for differential survival in association with morning vs afternoon dialysis include salutary effects of sleep in the morning or less efficient biochemical exchange during afternoon dialysis. Results from this cohort study may warrant prospective observational studies and randomized clinical trials that systematically alter the time of day at which HD is administered. PMID- 11730445 TI - School-associated violent deaths in the United States, 1994-1999. AB - CONTEXT: Despite the public alarm following a series of high-profile school shootings that occurred in the United States during the late 1990s, little is known about the actual incidence and characteristics of school-associated violent deaths. OBJECTIVE: To describe recent trends and features of school-associated violent deaths in the United States. DESIGN, SETTING, AND SUBJECTS: Population based surveillance study of data collected from media databases, state and local agencies, and police and school officials for July 1, 1994, through June 30, 1999. A case was defined as a homicide, suicide, legal intervention, or unintentional firearm-related death of a student or nonstudent in which the fatal injury occurred (1) on the campus of a public or private elementary or secondary school, (2) while the victim was on the way to or from such a school, or (3) while the victim was attending or traveling to or from an official school sponsored event. MAIN OUTCOME MEASURES: National estimates of risk of school associated violent death; national trends in school-associated violent deaths; common features of these events; and potential risk factors for perpetration and victimization. RESULTS: Between 1994 and 1999, 220 events resulting in 253 deaths were identified; 202 events involved 1 death and 18 involved multiple deaths (median, 2 deaths per multiple-victim event). Of the 220 events, 172 were homicides, 30 were suicides, 11 were homicide-suicides, 5 were legal intervention deaths, and 2 were unintentional firearm-related deaths. Students accounted for 172 (68.0%) of these deaths, resulting in an estimated average annual incidence of 0.068 per 100 000 students. Between 1992 and 1999, the rate of single-victim student homicides decreased significantly (P =.03); however, homicide rates for students killed in multiple-victim events increased (P =.047). Most events occurred around the start of the school day, the lunch period, or the end of the school day. For 120 (54.5%) of the incidents, respondents reported that a note, threat, or other action potentially indicating risk for violence occurred prior to the event. Homicide offenders were more likely than homicide victims to have expressed some form of suicidal behavior prior to the event (odds ratio [OR], 6.96; 95% confidence interval [CI], 1.96-24.65) and been bullied by their peers (OR, 2.57; 95% CI, 1.12-5.92). CONCLUSIONS: Although school-associated violent deaths remain rare events, they have occurred often enough to allow for the detection of patterns and the identification of potential risk factors. This information may help schools respond to this problem. PMID- 11730446 TI - Delirium in mechanically ventilated patients: validity and reliability of the confusion assessment method for the intensive care unit (CAM-ICU). AB - CONTEXT: Delirium is a common problem in the intensive care unit (ICU). Accurate diagnosis is limited by the difficulty of communicating with mechanically ventilated patients and by lack of a validated delirium instrument for use in the ICU. OBJECTIVES: To validate a delirium assessment instrument that uses standardized nonverbal assessments for mechanically ventilated patients and to determine the occurrence rate of delirium in such patients. DESIGN AND SETTING: Prospective cohort study testing the Confusion Assessment Method for ICU Patients (CAM-ICU) in the adult medical and coronary ICUs of a US university-based medical center. PARTICIPANTS: A total of 111 consecutive patients who were mechanically ventilated were enrolled from February 1, 2000, to July 15, 2000, of whom 96 (86.5%) were evaluable for the development of delirium and 15 (13.5%) were excluded because they remained comatose throughout the investigation. MAIN OUTCOME MEASURES: Occurrence rate of delirium and sensitivity, specificity, and interrater reliability of delirium assessments using the CAM-ICU, made daily by 2 critical care study nurses, compared with assessments by delirium experts using Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria. RESULTS: A total of 471 daily paired evaluations were completed. Compared with the reference standard for diagnosing delirium, 2 study nurses using the CAM-ICU had sensitivities of 100% and 93%, specificities of 98% and 100%, and high interrater reliability (kappa = 0.96; 95% confidence interval, 0.92-0.99). Interrater reliability measures across subgroup comparisons showed kappa values of 0.92 for those aged 65 years or older, 0.99 for those with suspected dementia, or 0.94 for those with Acute Physiology and Chronic Health Evaluation II scores at or above the median value of 23 (all P<.001). Comparing sensitivity and specificity between patient subgroups according to age, suspected dementia, or severity of illness showed no significant differences. The mean (SD) CAM-ICU administration time was 2 (1) minutes. Reference standard diagnoses of delirium, stupor, and coma occurred in 25.2%, 21.3%, and 28.5% of all observations, respectively. Delirium occurred in 80 (83.3%) patients during their ICU stay for a mean (SD) of 2.4 (1.6) days. Delirium was even present in 39.5% of alert or easily aroused patient observations by the reference standard and persisted in 10.4% of patients at hospital discharge. CONCLUSIONS: Delirium, a complication not currently monitored in the ICU setting, is extremely common in mechanically ventilated patients. The CAM-ICU appears to be rapid, valid, and reliable for diagnosing delirium in the ICU setting and may be a useful instrument for both clinical and research purposes. PMID- 11730447 TI - Large-scale quarantine following biological terrorism in the United States: scientific examination, logistic and legal limits, and possible consequences. AB - Concern for potential bioterrorist attacks causing mass casualties has increased recently. Particular attention has been paid to scenarios in which a biological agent capable of person-to-person transmission, such as smallpox, is intentionally released among civilians. Multiple public health interventions are possible to effect disease containment in this context. One disease control measure that has been regularly proposed in various settings is the imposition of large-scale or geographic quarantine on the potentially exposed population. Although large-scale quarantine has not been implemented in recent US history, it has been used on a small scale in biological hoaxes, and it has been invoked in federally sponsored bioterrorism exercises. This article reviews the scientific principles that are relevant to the likely effectiveness of quarantine, the logistic barriers to its implementation, legal issues that a large-scale quarantine raises, and possible adverse consequences that might result from quarantine action. Imposition of large-scale quarantine-compulsory sequestration of groups of possibly exposed persons or human confinement within certain geographic areas to prevent spread of contagious disease-should not be considered a primary public health strategy in most imaginable circumstances. In the majority of contexts, other less extreme public health actions are likely to be more effective and create fewer unintended adverse consequences than quarantine. Actions and areas for future research, policy development, and response planning efforts are provided. PMID- 11730448 TI - Ensuring patient safety in clinical trials for treatment of acute stroke. PMID- 11730449 TI - Patient selection for lung transplantation. PMID- 11730450 TI - Malnutrition and mortality in Kohistan District, Afghanistan, April 2001. AB - CONTEXT: The humanitarian situation in Afghanistan has been deteriorating for more than 3 years because of civil war and severe drought. Because of recent events, the international community is predicting a severe worsening of the country's current situation. OBJECTIVE: To assess the magnitude and causes of mortality and prevalence of malnutrition in Kohistan district, Faryab province, Afghanistan. DESIGN: Two-stage, 30-cluster household survey conducted April 4 through 10, 2001, which included anthropometric measurements, assessment of food coping mechanisms, and retrospective mortality data collection for November 26, 2000, through April 4, 2001. SETTING AND PARTICIPANTS: A total of 378 households comprising 3165 people living in Kohistan district, Faryab province, Afghanistan. MAIN OUTCOME MEASURES: Crude mortality rate, mortality rate among children younger than 5 years, causes of death, prevalence of wasting and stunting among children aged 6 to 59 months (weight-for-height and height-for-age z scores <-2, respectively), and food coping mechanisms. RESULTS: The crude mortality rate among the 3165 persons surveyed during the period of interest was 2.6 (95% confidence interval [CI], 1.7-3.5) per 10 000 per day and the mortality rate among 763 children younger than 5 years was 5.9 (95% CI, 2.0-8.8) per 10 000 per day. Diarrhea (25.0%), respiratory tract infections (19.4%), measles (15.7%), and scurvy (6.5%) caused most of the 108 deaths. The prevalences of wasting and stunting among 708 children aged 6 to 59 months were 7.0% (95% CI, 5.9%-9.0%) and 63.7% (95% CI, 58.6%-68.8%), respectively. Evidence of late-stage food coping mechanisms and prefamine indicators existed among the population. CONCLUSIONS: These data indicate that, by April 2001, a humanitarian crisis already existed in Kohistan. Essential humanitarian services, including food aid and public health programs, are urgently required in such regions of Afghanistan and will be crucial if a worsening humanitarian crisis is to be avoided. For these services and programs to be implemented, the international community needs to create adequate humanitarian space (ie, a secure and accessible location where humanitarian organizations can provide services to emergency-affected populations) to ensure that humanitarian organizations have access to populations within Afghanistan as well as to refugees who flee to surrounding countries. PMID- 11730452 TI - msJAMA: new directions in tobacco control. PMID- 11730453 TI - msJAMA: reducing smoking prevalence to 10% in five years. PMID- 11730454 TI - msJAMA: removing barriers to local tobacco regulation. PMID- 11730455 TI - msJAMA: the worldwide tobacco treaty. PMID- 11730456 TI - msJAMA: the necessity of global tobacco regulations. PMID- 11730480 TI - An associated process for the purification of immuno globulin G, catalase, superoxide dismutase and albumin from haemolysed human placenta blood. AB - The human placenta is a rich raw material for production of many biopharmaceutical products. Here we describe a co-purification process for the production of four different proteins from haemolysed human placenta blood: IgG, catalase (Cat), superoxide dismutase (Sod) and albumin (Alb). The process can be divided in two parts: the common steps and the specific separation techniques for each protein. The common steps are: extraction, haemoglobin precipitation, concentration/diafiltration and the first Q-Sepharose chromatography step. At this chromatography step the process is branched: while IgG and Cat were recovered in the flow-through, Sod and Alb were eluted separately. IgG and Cat were separated in a second Q-Sepharose chromatography step during which IgG was recovered in the flow-through, whereas Cat bound to the resin. IgG was purified by S-Sepharose chromatography, followed by selective precipitation with n octanoic acid, yielding about 0.4 g of IgG per kg of placenta. Cat was eluted at the second Q-Sepharose chromatography step and was purified by Blue Sepharose chromatography. A total of 1.8 x 10(6) units of Cat were recovered/kg of placenta, with a specific activity of 45000 units/mg of protein. Sod was further purified by S-Sepharose and Phenyl-Sepharose chromatography steps and recovered in the non-adsorbed fractions. The yield of Sod was 2.1 x 10(5) units/kg of placenta, with a specific activity of 1194 units/mg of protein. Alb purification was followed by a combined process including thermocoagulation and treatment with activated charcoal. The final step was Phenyl-Sepharose chromatography. The process yielded 3.1 g of Alb/kg of placenta. The described methodology was designed to be easily scaled-up for industrial production. PMID- 11730481 TI - In vitro phagocytosis of carrier mouse red blood cells is increased by Band 3 cross-linking or diamide treatment. AB - Chemical alteration of red blood cells (RBCs) can induce increased phagocytosis of modified cells by macrophages. In this study we have used different chemical treatments for the modification of the mouse red-blood-cell membrane surface, namely oxidant compounds, such as ascorbate/Fe(+2) and diamide [azodicarboxylic acid bis(dimethylamide)], or Band 3-cross-linking reagents. We monitored the phagocytosis of oxidized or Band 3-cross-linked mouse red blood cells by peritoneal macrophages. The extent of phagocytosis of RBCs is not affected by oxidation with ascorbate/Fe(3+), but it is increased (up to 10%) by oxidation with 2 mM diamide. Furthermore, phagocytosis is greatly increased (up to 40%) as a result of cross-linking with either of two Band 3 bifunctional reagents [bis(sulphosuccinimidyl) suberate (BS(3)) and 3,3'-dithiobis(sulphosuccinimidyl propionate) (DTSSP)]. To evaluate targeting towards macrophages of such modified RBCs for therapeutical purposes, we have determined the phagocytosis of Band 3 carrier RBCs loaded with carbonic anhydrase. In this case phagocytosis is high enough (25%) to deliver the enzyme into macrophages. We have also assayed the influence of serum components and IgG on the efficiency of phagocytosis and discuss the possible phagocytosis mechanisms. In the case of BS(3)-cross-linked carrier RBCs, phagocytosis is markedly enhanced (from 12% up to 25%) by serum components. This opens a way for therapeutic application of these carrier RBCs, with special relevance in short-term delivery to cells of the mononuclear phagocytic system. PMID- 11730482 TI - Identification of an import signal for, and the nuclear localization of, human lactoferrin. AB - Many different unique functions have been attributed to lactoferrin (Lf), including DNA and RNA binding, and transport into the nucleus, where Lf binds to specific sequences and activates transcription. A pentapeptide, Gly-Arg-Arg-Arg Arg, corresponding to a region of the N-terminal portion of human Lf rich in basic amino acids, was synthesized and its intracellular localization was investigated. Peptide internalization was assayed using the rhodaminated form of the same molecule. This N-terminal peptide sequence is able to be internalized within less than 10 min at concentration as low as 1 microM, and its intracellular localization is nuclear, mainly nucleolar. Similar behaviour was observed using peptides composed of either all l or d amino acids, the last one being a retro-inverse peptide. The internalization process does not involve an endocytotic pathway, since no inhibition of the uptake was observed at 4 degrees C. The kinetics of peptide internalization was also evaluated. The internalization properties of such a short Lf pentapeptide have been assayed for its ability to transport peptide nucleic acids (PNAs) inside cells in order to improve their efficacy. The abundant transmembrane transport and nuclear localization of the proposed peptide, deriving from hLf and, for the first time, identified as a nuclear localization signal, could be used as an alternative strategy to tackle the unsolved problem of intracellular accumulation of antisense and antigene drugs and for the development of new pharmacological tools. PMID- 11730483 TI - Efficient cryopreservation of dendritic cells transfected with cDNA of a tumour antigen for clinical application. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells of the immune system and are currently being investigated in clinical applications as cancer vaccines. An efficient cryopreservation method would greatly contribute to their use in clinical trials. We have established a method for freezing of DCs derived from peripheral blood mononuclear cells using the plasma expander Gelifundol. This enabled us to reduce the concentration of the toxic DMSO to 5%. The method could be performed without the addition of fetal calf serum or any other serum. After freezing, the viability of the DCs was 90%. The cells exhibited all the phenotypic characteristics (CD11c+, HLA-DR+, CD80+, CD83+, CD86+) of DCs, as tested by flow cytometry. Cells transfected with cDNA for the tumour antigen mucin expressed this protein on their surfaces in the same manner as before freezing. The stimulating capacity of a mixed lymphocyte culture was also preserved. These findings offer an efficient method for the cryopreservation of DCs for use in clinical trials. PMID- 11730484 TI - Cold lability of lactate dehydrogenase isoenzymes and the effective preparation of reference material for clinical laboratory use. AB - To evaluate the storage of samples and enzyme reference materials, and to improve the commutability for inter-laboratory surveillance of activity values of lactate dehydrogenase (LD; EC 1.1.1.27) in clinical laboratory medicine and in animal veterinary medicine, we studied the electrophoretic patterns and cold lability of LD isoenzymes from tissue sources of some common vertebrate species and also from human serum sources. Among many isoenzymes from these sources, only rat LD fractions showed similar electrophoretic patterns to those of human sera, and rat LD-1 fraction was relatively cold-stable. Total LD and isoenzyme LD-1 activities in routine laboratory samples and quality-control sera were measured using eight kinds of commercially available LD assay kit, including lactate and pyruvate substrate systems. Coefficients of variation between these assay kits were markedly reduced when the activities were calculated using the partially purified rat LD-1 fraction as an enzyme reference material, compared with the activities calculated using the factor indicated in each assay kit. In the regression analysis, the intercept and slope were calculated for the regression equations obtained from 12 pairs of these assay kits. The values obtained from a small amount of human serum and control serum samples were within the 95% confidence regions of those from larger amounts of human serum samples by using the present rat LD-1 standard for measuring LD-1 activities with lactate substrate. It was evident that a cold-stable and homogeneous LD isoenzyme as an enzyme reference material might contribute to accurate measurement of activities in heterogeneous samples for inter-laboratory quality-assurance surveys in both human and animal clinical laboratory use. PMID- 11730485 TI - A simple method for the two-step preparation of two pure haemorphins from a total haemoglobin peptic hydrolysate by conventional low-pressure chromatographies. AB - The development of a simple purification method for two haemorphins, VV haemorphin-7 (Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe) and VV-haemorphin-4 (Val-Val Tyr-Pro-Trp-Thr), from a total peptic haemoglobin hydrolysate is described. Cation-exchange chromatography on CM-Sephadex was used as a pre-fractionation step for the hydrolysate. VV-haemorphin-7 and VV-haemorphin-4 were eluted in two different fractions. The second and final purification step was hydrophobic interaction chromatography on phenyl-Sepharose, which allowed the isolation of the two pure haemorphins. Haemoglobin and globin hydrolysates were compared as starting materials. For easy recovery of haemorphins and easy adjustment of conditions for final purification, a volatile buffer, ammonium acetate buffer, pH 6.5, was employed. This process, which allowed the preparation of pure haemorphins from a total protein hydrolysate, could be scaled up and used in the food industry. PMID- 11730486 TI - Optimization of the anti-(human CD3) immunotoxin DT389-scFv(UCHT1) N-terminal sequence to yield a homogeneous protein. AB - The production and regulatory approval processes for biopharmaceuticals require detailed characterization of potential products. Therapeutic proteins should preferably be homogeneous, although limited, reproducible, heterogeneity may be tolerated. A diphtheria toxin-based anti-(human CD3) immunotoxin, DT389 scFv(UCHT1), was expressed in Escherichia coli and purified following refolding [DT389 corresponds to amino acids 1-389 of diphtheria toxin, scFv is single-chain variable-region antibody fragment and UCHT1is an anti-(human CD3) monoclonal antibody]. Biochemical characterization of this molecule by MS and N-terminal sequencing by Edman degradation revealed that the protein was heterogeneous at the N-terminus, containing species both with (60%) and without (40%) the initiator methionine residue. In an attempt to generate an N-terminally homogeneous molecule, a panel of seven N-terminal variants was designed, based on the published specificity of bacterial methionine aminopeptidase. Following bacterial expression, partial purification and separation on SDS/PAGE, these proteins were subjected to N-terminal sequencing by Edman degradation. Three of the mutants yielded a 100% homogeneous amino acid sequence. By contrast, the original DT389-scFv(UCHT1) protein and four variant proteins yielded two sequences with varying ratios corresponding to species with and without methionine. The N-terminal sequences of the three homogeneous clones were MLADD and MLDD, where the methionine was completely retained, and SADD, where the methionine was completely removed. One of the homogeneous mutants (SADD) was expressed, refolded and purified and found to be equipotent with the parent immunotoxin. Thus, using a rational mutagenesis approach, three N-terminally homogeneous variants of DT389-scFv(UCHT1) have been identified, at least one of which is functionally indistinguishable from the parent immunotoxin. This approach is generally applicable to biopharmaceutical production and immunotoxin development in particular. PMID- 11730487 TI - Transphosphatidylation by immobilized phospholipase D in aqueous media. AB - Phospholipase D (PLD) from Streptomyces sp. was immobilized by covalent binding to aminopropyl-glass activated by glutardialdehyde and to the macroporous synthetic polymer VA-Epoxy Biosynth (from Riedel-de Haen, Seelze, Germany) pre activated by epoxy groups. The immobilized PLDs were examined for the synthesis of phosphatidylglycerol (PG) from soybean (Glycine max) phosphatidylcholine (PC) in purely aqueous solutions in comparison with commonly used diethyl ether/buffer systems. In contrast with general assumptions, the transphosphatidylation was shown to yield a high percentage of PG, even in pure buffer. With PLD immobilized to VA-Epoxy Biosynth, the formation of phosphatidic acid (PA) is insignificant, while the yield of PG amounts to 60%. With PLD immobilized to porous glass (average pore diameter 17 nm), higher yields of PG (72%) are reached, but the formation of PA also increases (up to 10%). In comparison with the reaction in the diethyl ether/buffer system, however, the conversion of PC into PG proceeds much more slowly. Detergents such as Triton X-100 accelerate the reaction. PMID- 11730488 TI - Identification of the ligand-binding domain of human vascular-endothelial-growth factor receptor Flt-1. AB - The vascular-endothelial-growth-factor (VEGF) receptor Flt-1 has been shown to be involved in vasculogenesis and angiogenesis. The receptor is characterized by seven immunoglobulin-like loops within the extracellular domain and the first three N-terminal immunoglobulin-like loops are involved in high-affinity binding of VEGF. The minimal extracellular domains of Flt-1 to achieve VEGF binding were screened using the yeast two-hybrid system. The result showed that the binding capacity of loop2-3 was close to that of loop1-3. The two truncated mutants consisting of loop2-3 and loop1-3 were expressed in the methylotrophic yeast Pichia pastoris at high levels (0.3 g[corrected]/litre). The corresponding proteins, named soluble (s)Flt-1(2-3) and sFlt-1(1-3), were purified. An in vitro biological activity assay showed that the binding capacity of sFlt-1(2-3) to human VEGF(165) and the inhibiting effect of it on human umbilical-vein endothelial cell proliferation stimulated by human VEGF(165) were close to those of sFlt-1(1-3). Animal tests showed that sFlt-1(2-3) could significantly inhibit the formation of regenerated blood vessels stimulated by hVEGF(165). PMID- 11730489 TI - Studies on the inactivation of soluble and immobilized papain by the ascorbic acid-Cu2+ system: a model to propose the effect of free radicals on membrane bound enzymes in vivo. AB - Free radicals have been suggested to be widely implicated as the species responsible for harmful biological processes, such as aging, carcinogenesis and numerous other diseases. The mechanism of biological damage produced in such processes has been investigated in a wide variety of systems, including studies on proteins, enzymes, nucleic acids and carbohydrates. In the present study we selected an ascorbic acid-transition-metal ion (ASA-Cu(2+)) system in order to understand the mechanism of soluble and membrane-bound enzyme inactivation by generating free radicals. Papain, a thiol protease, was immobilized on an immobilized-metal-ion carrier and used as a model to examine the inactivation behaviour of membrane-bound enzymes. A comparison was made between the inactivation of soluble and immobilized papain by free radicals, and the potential of different radical scavengers to prevent the inactivation of enzyme was examined. PMID- 11730490 TI - Plasma levels of thrombin-antithrombin complexes predict preterm premature rupture of the fetal membranes. AB - OBJECTIVE: Decidual hemorrhage (abruption) is strongly associated with preterm premature rupture of fetal membranes (PPROM). Moreover, thrombin enhances decidual matrix metalloproteinase (MMP) expression, and MMP has been strongly linked to PPROM. The current study sought to determine whether increased thrombin activation, as assessed by circulating maternal plasma thrombin-antithrombin (TAT) complexes, predicted subsequent PPROM. STUDY DESIGN: We conducted a nested, case-control study of plasma TAT levels, measured by sensitive immunoassay, among 27 women with a singleton preterm birth preceded by PPROM and 54 matched, term controls. Receiver operating characteristic curve analysis was performed to identify the optimal TAT cut-off level predicting PPROM. RESULTS: Mean gestational age at delivery in cases was 33.3 weeks, compared to 39.7 weeks in controls (p < 0.001). Compared with controls, women with PPROM had increased median plasma TAT levels in both the second trimester (5.1 microg/l (range 2.2 26.3 microg/l) vs. 3.2 microg/l (range 1.3-7.3 microg/l); p = 0.001) and third trimester (7.0 microg/l (range 2.6-85.8 microg/dl) vs. 4.8 microg/l (range 1.7 15.4 microg/dl); p = 0.01). In the PPROM group, 16.0% of the women exhibited bleeding during the pregnancy, while the corresponding value among controls was 3.6% (p = 0.07). In the second trimester, the odds ratio for PPROM with a TAT level of > 3.9 microg/l was 6.0 (95% CI 1.67-21.1). This value predicted PPROM with a sensitivity of 88%, specificity of 68% and positive and negative predictive values of 82% and 97%, respectively. CONCLUSION: Second-trimester elevated plasma TAT concentrations are predictive of subsequent PPROM. These data provide further evidence that PPROM is associated with decidual thrombin activation. PMID- 11730491 TI - Outcome of infants delivered between 24 and 28 weeks' gestation in women with severe pre-eclampsia. AB - OBJECTIVE: To determine whether there are differences in neonatal outcome between infants born to mothers with severe pre-eclampsia and those born to normotensive mothers with preterm labor and intact membranes between 24 and 28 weeks' gestation. MATERIALS AND METHODS: Over a 4-year period between 1991 and 1995, neonates of women with severe pre-eclampsia delivering between 24 and 28 weeks were matched for maternal age, antenatally assigned gestational age and mode of delivery to normotensive women delivering during the same period. RESULTS: Fifty eight women with severe pre-eclampsia were matched to 58 normotensive controls who delivered as a result of preterm labor. Antenatal steroids were used more often in pre-eclamptic women (75% vs. 47%, p < 0.01). The mean birth weight of pre-eclamptic neonates was significantly lower than that of controls, 767 g vs. 989 g, respectively. Other neonatal complications were similar for both groups. Neonates of pre-eclamptics required longer ventilator support (21 vs. 16 median days, p = 0.03). Neonatal survival was similar for both groups (72% and 79% for pre-eclamptics and normotensives, respectively). CONCLUSIONS: Neonates born to patients with severe pre-eclampsia have similar survival but a lower birth weight and require longer ventilator support than neonates born to women with preterm labor. PMID- 11730492 TI - Labor induction for the preterm severe pre-eclamptic patient: is it worth the effort? AB - OBJECTIVE: The purpose of this study was to examine the success rate of labor induction in patients with severe pre-eclampsia delivered at < or = 34 weeks' gestation; to identify factors associated with its success; and to evaluate neonatal outcomes based on induction success or failure. METHODS: We identified pregnancies complicated by severe pre-eclampsia delivered at < or = 34 weeks' at our institution from 1991 to 1998. Women who underwent labor induction and had successful vaginal delivery were compared to those who underwent labor induction, but required Cesarean delivery. Multiple logistic regression analyses were performed to assess factors associated with successful induction and neonatal outcome. RESULTS: Over the 7-year study period, there were 215 patients meeting the criteria. Sixty-four (29.8%) did not undergo a labor attempt; 69 of 151 (46%) women who underwent labor induction achieved vaginal delivery. Labor induction was successful in 0%, 6.6%, 35.3% and 68.5% of cases at 24-26, 27-28, 29-31 and 32-34 weeks' gestation, respectively. By logistic regression the only factor positively associated with successful induction was gestational age at delivery (p = 0.001), while induction for non-reassuring fetal testing was inversely associated (p = 0.02). Induction attempt, failed induction and delivery mode were not associated with increased neonatal morbidity. CONCLUSIONS: In women with severe pre-eclampsia remote from term, attempted labor induction did not appear to increase neonatal morbidity, but was rarely successful at < 28 weeks. PMID- 11730493 TI - Appropriateness of antibiotic use in the postpartum period. AB - OBJECTIVE: To determine the appropriateness of current postpartum antibiotic use in clinical practice. METHODS: Medical records were reviewed for all patients delivering in a 3-month period who received postpartum antibiotics during the delivery hospitalization. Subjects were excluded if they received a single postpartum antibiotic dose as part of a mitral valve prolapse prophylaxis protocol, or if they received no more than one postpartum antibiotic dose for surgical prophylaxis. Characteristics of postpartum antibiotic use were abstracted. RESULTS: Two hundred and eleven of 1537 (14%) delivering patients met the inclusion criteria. Seventy-four (35%) delivered vaginally and 137 (65%) delivered by Cesarean section. Postpartum fevers were found in 40 (54%) of vaginal delivery cases and 80 (58%) of women delivering by Cesarean section who received postpartum antibiotics (p = 0.54). For vaginal deliveries there were no differences in the duration of antibiotic use or number of antibiotic doses based on fever status. For Cesarean deliveries, a fever was associated with more antibiotic doses and a longer duration of antibiotic use. Physician justification for antibiotic use was documented in only 116 cases (55%). CONCLUSIONS: The high proportion of women receiving postpartum antibiotics having no evidence for infection or documented indication for therapy suggests that antibiotics may not be appropriately used in the postpartum period. PMID- 11730494 TI - Does uterine wiping influence the rate of post-Cesarean endometritis? AB - OBJECTIVE: To compare two methods of uterine cleansing to remove residual amnion chorion at the time of Cesarean delivery. METHODS: All consenting uninfected parturients undergoing a non-emergency Cesarean delivery at the university were enrolled. All patients received antibiotic prophylaxis of a 1-g intravenous bolus of a first-generation cephalosporin immediately after cord clamping. Postpartum endometritis was defined as a temperature of > or = 100.4 degrees F (38 degrees C) on two occasions 6 h apart after the first 24 h with uterine tenderness and/or foul-smelling lochia. RESULTS: Between January 1998 and February 2000 there were 1400 patients enrolled in this investigation. A total of 153 women were determined to have chorioamnionitis present at surgery and 17 women refused participation, leaving 614 women in the uterine wipe group and 616 in the no wipe group. Maternal demographics were similar between patient groups (NS). Intrapartum factors including gestational age at time of Cesarean section, type of anesthesia used for the operative delivery, hours of amnion rupture prior to delivery, use of internal monitoring devices, type of skin incision, method of placental removal, total operative time and estimated blood loss were similar between patient groups (NS). An almost identical number of patients in each arm of the study developed post-Cesarean endometritis (65 out of 614, 10.5%, in the uterine wipe group vs. 66 out of 616, 10.7%, in the no wipe group; p = 1). CONCLUSIONS: Both methods of removing residual amnion-chorion membrane tissue and associated debris at surgery were comparably effective, but neither produced a dramatic reduction in the incidence of postoperative endometritis. PMID- 11730495 TI - The combination of computerized cardiotocography and amniotic fluid index for the prediction of neonatal acidemia at birth: a modified biophysical profile. AB - OBJECTIVE: To study the combination of computerized cardiotocography (cCTG) and the amniotic fluid index (AFI) in the prediction of neonatal acidemia at birth. METHODS: A total of 89 singleton third-trimester high-risk pregnancies delivered by Cesarean section, with an AFI evaluated within 24 h from birth, and an antepartum cCTG performed within 6 h from delivery, were studied. The score was the sum of values for AFI (oligo/anhydramnios = 1, normal = 0) and cCTG (Dawes Redman criteria, not met = 1, met = 0). The endpoint was to predict an abnormal neonatal outcome as defined by an umbilical artery pH of < or = 7.2. RESULTS: Fifteen neonates had an umbilical artery pH of < 7.2. The combination of cCTG + AFI score was able to predict pH values (< or = 7.20) with an OR = 2.83 (p < 0.02). The diagnostic accuracy of the combination of cCTG + AFI was as follows: sensitivity 80%, specificity 58%, positive predictive value 28%, negative predictive value 83%. COMMENT: We suggest that the cCTG + AFI score may be of value in the prediction of neonatal acidemia and help in the management of third trimester high-risk pregnancies. PMID- 11730496 TI - Oxytocin use in grand-multiparous patients: safety and complications. AB - OBJECTIVE: To determine whether the use of oxytocin for the augmentation of labor in grandmultiparous women increases the risk of peripartum complications. STUDY DESIGN: During the years 1989-97, 11 075 grand-multiparous women delivered at our institution. In 424 grand-multiparous women, intravenous oxytocin was used for augmentation of labor. The control group consisted of the other 10 651 grand multiparous women. All women were monitored for fetal heart rate and uterine contractions. We compared the rates of maternal and perinatal complications in these two groups by using chi(2) analysis and Fisher's exact test when appropriate. RESULTS: No significant differences were found between the oxytocin and the control groups in the rates of placental abruption, intrapartum fetal death, postpartum hemorrhage, uterine rupture, fetal distress, meconium-stained amniotic fluid, an Apgar score of less than 7 at 5 min, Cesarean section, retained placenta and vaginal and cervical lacerations. In contrast, a significant increase in the rate of vacuum deliveries was observed in patients given oxytocin as compared to controls (3.5% vs. 1.4%, respectively; p = 0.001). CONCLUSIONS: The use of oxytocin in the grand-multiparous parturient was a safe procedure with no significant increase in peripartum complications. However, a higher rate of vacuum deliveries was found. PMID- 11730497 TI - The effect of a pill inserter on vaginal misoprostol dosing. AB - OBJECTIVE: To determine whether the method of placing a 25-microg misoprostol chip into the posterior fornix to achieve cervical ripening affects the drug's efficacy. METHODS: A pill inserter was used to place a misoprostol chip into the posterior fornix for the purpose of cervical ripening prior to induction of labor. Data from a control group were obtained by retrospective chart review. RESULTS: The control and study groups contained 49 patients each. Compared to placing the misoprostol chip with a lubricated finger, the use of the pill inserter resulted in statistically significantly more patients receiving only one dose. This occurred either because a Bishop score of 8 or greater was achieved or because repeat dosing was disallowed secondary to the onset of uterine contractions. Although the total number of patients subsequently requiring oxytocin was significantly increased, there was no difference in the use of oxytocin for either induction or augmentation of labor. The lengths of the latent and active phases of labor did not differ between the two groups. CONCLUSION: The number of doses of a 25-microg misoprostol chip for cervical ripening that result in uterine contractions, with or without a change in the Bishop score, is affected by the method used to place it in the vagina. PMID- 11730498 TI - Ovine intramembranous pathway permeability: use of solute clearance to determine membrane porosity. AB - OBJECTIVE: The contribution of the fetal chorioamniotic membranes (i.e. the intramembranous pathway) to the regulation and maintenance of amniotic fluid (AF) volume and composition has yet to be completely understood. Knowledge of membrane permeability properties is vital to understanding how the intramembranous pathway contributes to the overall maintenance of AF homeostasis. Although there are significant data regarding the regulation of intramembranous water flow, there is little understanding of the regulation of intramembranous solute flow. In the present study, we sought to determine the effect of molecular weight or size of non-polar compounds on intramembranous solute movement in the ovine model. METHODS: Five singleton ovine fetuses (117 +/- 3 days) were chronically prepared with bladder, tracheal, amniotic cavity and femoral arterial and venous catheters and an esophageal occluder. The allantoic membranes were excised. After 5 days' recovery, AF volume was calculated by intraamniotic injection of 99Tc-labelled red blood cells (time -6 to 0 h). At time 0, AF exchange routes were limited to the intramembranous pathway by inflation of the esophageal occluder and external drainage of fetal urine and lung fluid. Following intra-amniotic injection of creatinine (Cr, 1 g, MW 11 000 Da, 4 A) and [125I]albumin (RISA, 250 microCi, MW 69 000 Da, 36 A), maternal and fetal plasma and AF samples were collected at timed intervals during the subsequent 5 h. AF solute clearance (Cl(x)) was determined by the changes in AF total solute content. RESULTS: Cr and RISA disappeared from the AF with a corresponding increase in fetal, though not maternal, plasma levels. The mean Cl(Cr) was significantly greater than Cl(RISA) (2.0 +/- 0.3 ml/min vs. 1.0 +/- 0.2 ml/min; p < 0.04). CONCLUSION: Solute clearance from the amniotic cavity is inversely proportional to solute molecular weight/size. Although the membrane comprising the ovine intramembranous pathway is size restrictive, membrane pores allow passage of non-polar solutes up to 36 A. Knowledge of membrane permeability characteristics is essential for the utilization of the intramembranous pathway for fetal therapeutics. PMID- 11730499 TI - Absence of physiological tolerance to cocaine in pregnant sheep. AB - OBJECTIVE: To determine whether repetitive administration of cocaine to sheep during pregnancy altered basal hemodynamic states in the mother and fetus, and to determine whether this cocaine exposure would alter subsequent hemodynamic responses to cocaine. METHODS: Cocaine or saline was administered to 16 pregnant sheep daily from day 75 to day 128 of gestation (term = 145 days). At 128 days' gestation, maternal and fetal basal physiological measurements, including organ specific blood flow in the fetus, were determined. Each experimental and control ewe then received cocaine 2 mg/kg and these physiological parameters were again measured over the next 30 min to determine whether the experimental animals had developed tolerance to the effects of cocaine. RESULTS: No differences were seen in basal physiological parameters between treatment groups. Likewise, following an acute administration of cocaine, physiological parameters in both groups responded in a similar fashion. Fetal hypoxemia occurred in both groups after the ewe received cocaine. In response to hypoxemia, whether it was the animals' first or 53rd exposure to cocaine, fetal cerebral, myocardial and adrenal blood flow increased so that oxygen delivery was unimpaired. CONCLUSIONS: For the cardiovascular parameters measured in this study, we found no tolerance to cocaine in the ewe or fetus. The acute hemodynamic effects of maternal cocaine administration were as severe for animals having received it more than 50 times as for those that received it for the first time. PMID- 11730500 TI - Fetal heart rate patterns in normal and ritodrine-treated pregnancies, detected by magnetocardiography. AB - OBJECTIVE: The aim of the present study was to test the validity of magnetocardiography in the diagnosis of fetal heart rate arrhythmias in normal pregnancies, as compared to the number of arrhythmias reported in other series, which were detected by use of other diagnostic techniques. We also evaluated the influence of ritodrine on the fetal heart rhythm in pregnancies treated for the risk of preterm labor by means of magnetocardiography, in order to provide preliminary results that could be utilized in the future establishment of magnetocardiography as a screening procedure in the diagnosis and management of fetal arrhythmias. METHODS: We performed a prospective study on two subgroups of pregnant women: one of 84 women with normal healthy singleton pregnancies and one of 68 pregnant women treated with ritodrine for the risk of preterm labor. RESULTS: The prevalence of fetal arrhythmias in the first subgroup was 3.5% (3/84), while in the second subgroup the prevalence was 16% (11/68). CONCLUSIONS: The incidence of fetal arrhythmias detected in our population of normal pregnancies was comparable to that reported in previous studies by use of other techniques. Results gained from the second subgroup, although not comparable to others, owing to lack of similar reports, led us to believe that magnetocardiography's advantages over conventional methods of fetal cardiac surveillance could highlight the technique as a useful screening procedure for the detection of preterm fetuses, which should be submitted to closer investigation, because of the arrhythmias caused by ritodrine infusion. PMID- 11730501 TI - Contraindication of magnesium sulfate in a pregnancy complicated with late-onset diabetes mellitus and sensory deafness due to mitochondrial myopathy. AB - A primipara affected by late-onset diabetes and sensory deafness because of mitochondrial myopathy was hospitalized for threatened preterm delivery. Magnesium sulfate was started for tocolysis, resulting in general muscle damage, although the mitochondrial myopathy did not deteriorate during pregnancy. Magnesium sulfate may be contraindicated in pregnancy with mitochondrial myopathy. PMID- 11730503 TI - Post-splenectomy splenic artery aneurysm rupture in an atypical presentation of pre-eclampsia. AB - Splenic artery aneurysm rupture in pregnancy is an uncommon catastrophic event. We report a patient who presented at 15 3/7 weeks with atypical pre-eclampsia. After termination was recommended, the patient chose to continue the pregnancy. Reversal of clinical and laboratory abnormalities occurred and the patient was discharged. The patient presented again at 24 weeks with severe pre-eclampsia and residual splenic artery aneurysm rupture, at the site of a splenectomy that had been performed 24 years previously. PMID- 11730502 TI - Antenatal diagnosis of mirror-image dextrocardia in association with situs inversus and Turner's mosaicism. AB - We describe the antenatal diagnosis of a fetus with mirror-image dextrocardia, complete situs inversus and Turner's mosaicism (45,XO/46,XY) that was artificially terminated at 19 weeks. Autopsy confirmed our initial findings. This case represents an unusual combination of anomalies rarely encountered in clinical practice. PMID- 11730504 TI - Choledochal cysts diagnosed in pregnancy: a case report and review of treatment options. AB - An 18-year-old woman at 18 weeks' gestation presented with abdominal pain and jaundice. After extensive evaluation, she was diagnosed as having a choledochal cyst. Ten days after percutaneous drainage, she underwent complete excision and Roux-en-Y hepaticojejunostomy anastomosis. At term, she delivered vaginally without complications. Although choledochal cysts are rare in pregnancy, it is important for the obstetrician to be familiar with the presentation and the best treatment modalities available. Delay in diagnosis and inappropriate treatment can result in maternal morbidity and fetal mortality. PMID- 11730515 TI - The Peril of the Pearl: In Its Strength Lay Its Vulnerability. PMID- 11730516 TI - Advances in Palliative Care in Latin America and the Caribbean: Ongoing Projects of the Pan American Health Organization (PAHO). AB - The Pan American Health Organization (PAHO) is engaged in field demonstration projects in Latin America and the Caribbean to expand the availability and effectiveness of palliative care services. Workshops and calls for proposals address the multiple causes of inadequate palliative care services that are typical for countries in the developing world. These include regulatory barriers and drug interdiction policies that interfere with the medical use of opioids, lack of education among health professionals, and insufficient palliative care infrastructure. PMID- 11730518 TI - Innovations in End-of-Life Care. PMID- 11730517 TI - Recent Literature. PMID- 11730519 TI - Session Abstracts. PMID- 11730521 TI - Corrections. PMID- 11730522 TI - A Few Words. PMID- 11730524 TI - What "Stage" Can You Use as a Platform to Teach End of Life? PMID- 11730523 TI - Margaret's Treasure. PMID- 11730526 TI - Recent Literature. PMID- 11730525 TI - Politics, Palliation, and Canadian Progress in End-of-Life Care. AB - How do health care consumers, private sector organizations, professionals, and government agencies work toward ensuring quality end-of-life care? This challenge is by no means uniquely Canadian; our approach may thus have applications that extend well beyond our national borders. Raising the profile of palliative care in Canada has occurred on many fronts. Local, provincial, and national efforts in end-of-life care have long been underway. The Canadian Palliative Care Association has been particularly active, recently organizing various stakeholder groups to develop a strong national advocacy partnership for quality end-of-life care. Perhaps most distinctive has been the involvement and nationwide visibility of Canada's Senate. Their report, Quality End-of-Life Care: The Right of Every Canadian has seen Canadian palliative care gain unprecedented momentum. The federal government recently took the unprecedented step of creating a new office to represent palliative care within the federal cabinet. In Canada, it would appear that palliative care is an idea whose time has come. PMID- 11730527 TI - Innovations in End-of-Life Care. PMID- 11730553 TI - Role of calcitonin gene-related peptide in prostaglandins-mediated ischemic preconditioning in guinea pig hearts. AB - AIM: To examine the role of calcitonin gene-related peptide (CGRP) in ischemic preconditioning induced by prostaglandins in isolated guinea pig hearts. METHODS: The isolated guinea pig hearts were perfused in a Langendorff model. The heart rate, coronary flow, left ventricular pressure, and its first derivatives (+/ dp/dt(max)) were recorded and the calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) and 6-keto-PGF(1 alpha) were measured. RESULTS: Endothelin-1 (200 pmol in 1 mL K-H buffer) reduced the left ventricular developed pressure and its first derivatives (+/-dp/dt(max)), heart rate, and coronary flow. Preconditioning with two cycles of 5-min global ischemia and 5-min reperfusion attenuated endothelin-1 induced myocardial injury, and concentrations of both CGRP and 6-keto-PGF(1alpha) in the coronary effluent were markedly raised in the preconditioning periods. Pretreatment with capsaicin, which depletes endogenous CGRP, abolished the elevated level of CGRP concomitantly with loss of the cardioprotection induced by ischemic preconditioning. CGRP(8-37) (100 nmol/L), a selective CGRP1 receptor antagonist, also abolished the protective effects of ischemic preconditioning. After pretreatment with indometacin (10 micromol/L), an inhibitor of cyclooxygenase, the protective effects of ischemic preconditioning were abolished and the release of 6-keto-PGF1alpha was no longer elevated. Pretreatment with indometacin abolished the elevated level of CGRP in the coronary effluent. CONCLUSION: Endogenous prostaglandins are involved in the protective effects of ischemic preconditioning, and the beneficial effects of prostaglandins are mediated by CGRP in the guinea pig heart. PMID- 11730554 TI - Stimulation by melittin of Na+-Ca2+ exchange current in ventricular myocytes of guinea pigs. AB - AIM: To study the mechanism of calcium overload induced by melittin in myocytes. METHODS: Whole cell patch-clamp technique was applied for recording the currents. RESULTS: Mel 0.05, 0.1 micromol/L increased the peak amplitude of I(Na) (nA) from -2.1+/-0.8 to -3.2+/-1.0 (n=7, P < 0.05) and -3.7+/-1.5 (n=7, P < 0.05) respectively at testing potential of -40 mV. Mel 0.05, 0.1, 0.2 micromol/L had no significant effect on I(Ca), but enhanced I(Na-Ca) (pA) from 53+/-21 to 427+/-256 (n=5, P < 0.05), 349+/-147 (n=5, P<0.01) and 320+/-97 (n=5, P < 0.05) respectively at a testing potential of +50 mV. CONCLUSION: The stimulating effect of Mel on I(Na-Ca) rather than the effect on I(Ca) contributes to the calcium overload of myocytes. PMID- 11730555 TI - Inhibitory effect of recombinant TGFalpha-PE40 on neointimal proliferation after arterial balloon injury. AB - AIM: To investigate inhibitory effects of recombinant transforming growth factor alpha-Pseudomonas exotoxin 40 fusion protein (TGFalpha-PE40; TP40) on neointimal proliferation after arterial balloon injury. METHODS: Forty male rabbits fed a cholesterol rich diet were randomly divided into TP40 15 microg, 30 microg, 60 microg, physiologic saline control, and normal artery groups (n=8). Rabbits in the treatment groups were treated by local administration of TP40 (15 microg, 30 microg, and 60 microg per rabbit) 24 h postinjury, and those in the control group were treated by physiologic saline 24 h postinjury. Remained 8 rabbits in normal artery group were treated by TP40 (60 microg). Optical microscope, electron microscope, and computer image analysis were used to study arterial segments 2 weeks after treatment. RESULTS: Irregular thickening of the arterial intima, large amounts of smooth muscle cells (SMC) within the neointima, and stenosis of the arterial cavity were observed in the physiologic saline control group. Great inhibition of intimal proliferation and prevention of stenosis of the arterial cavity were observed in the TP40 treated groups that were examined 2 weeks postinjury by optical microscope. The uninjured carotids were histologically normal. Lots of destructural and necrotic SMC were observed in media in TP40 60 microg group by electron microscope. Computer image analysis showed that the neointimal area of the TP40-treated groups was markedly smaller than that of the saline control group (P < 0.01). CONCLUSION: Recombinant TP40 greatly inhibited neointimal proliferation after arterial balloon injury. PMID- 11730556 TI - Uridine triphosphate prolongs action potential duration of guinea pig papillary muscles via P2Y2 purinoceptors. AB - AIM: To study the electrophysiologic effects of uridine triphosphate (UTP) on the guinea pig papillary muscles in vitro and purinoceptors related with the action of UTP. METHODS: Intracellular microelectrode method was used to record action potentials (AP) in guinea pig papillary muscles. RESULTS: UTP, adenosine triphosphate (ATP), and adenosine diphosphate (ADP) prolonged the action potential duration (APD) concentration dependently in guinea pig papillary muscles. The potency order was UTP=ATP > ADP. There was cross desensitization between the response to ATP and that to UTP, and neither Ado nor alpha, beta MeATP caused great change in AP of the papillary muscles. The prolongation of APD by UTP was not affected by sustained perfusion with aminophylline. As an osmotic pressure control equivalent to UTP 3 mmol/L, ceftriaxonum 3 mmol/L or NaCl 9 mmol/L induced a marked but slight prolongation of APD. CONCLUSION: UTP produced APD prolongation through specific and nonspecific actions, and the specific response to UTP was mediated by P2Y2 purinoceptors. PMID- 11730557 TI - Effect of cathepsin B on thymocyte apoptosis in spontaneously hypertensive rats. AB - AIM: To further investigate the mechanism of the thymus dysfunction in spontaneously hypertensive rats. METHODS: The location of cathepsin B expression was determined by in situ hybridization. The expression of cathepsin B was analyzed by Northern blot, and thymocyte apoptosis was detected by TUNEL and flow cytometer, respectively. RESULTS: The expression of cathepsin B in thymus paralleled with thymocyte apoptosis both in vitro and in vivo. Cathepsin B transcripts were located in the plasma of thymocytes and were more over-expressed of 6- and 8-week-old SHR than those in the thymus of WKY. CONCLUSION: Thymocyte apoptosis in SHR is increased and this phenomenon is associated with the expression of cathepsin B. PMID- 11730558 TI - Frequency dependent prolongation of effective refractory period by a complex class III antiarrhythmic agent CPU-86017. AB - AIM: To compare the prolongation of the effective refractory period (ERP) of a novel complex Class III agent CPU-86017 with compounds blocking INa (lorcainide, Lor), IK (dofetilide, Dof), ICa (verapamil, Ver) and a complex Class III multiple channel blocking agent (amiodarone, Ami) at different frequency levels. METHODS: The ERP of the guinea pig right papillary muscle and the anoxic left atrial muscle treated with high K+ at pH 6.6-6.8, in concentrations ranging 0.03-30 micromol/L was compared at frequency levels ranging from 0.5-4.0 Hz. RESULTS: CPU 86017 showed a positive frequency dependence (PFD) with respect to ERP in the papillary muscles and a mild reverse frequency dependence (RFD) in the diseased atrium. The potency of ERP prolonging effect of various agents at 4.0 Hz was Dof > CPU-86017 > Ver > Ami > Lor, and Dof > CPU-86017 > Ami > Lor > Ver in the normal papillary and diseased atrial muscle, respectively. CONCLUSION: The profile and potency of prolonging the ERP by CPU-86017 is similar to Dof which blocks IKr in the diseased atrium, and to Ver which blocks ICa in guinea pig ventricle. PMID- 11730559 TI - Cholinesterase inhibition by aluminium phosphide poisoning in rats and effects of atropine and pralidoxime chloride. AB - AIM: To investigate the cholinesterase inhibition and effect of atropine and pralidoxime (PAM) treatment on the survival time in the rat model of aluminium phosphide (AlP) poisoning. METHODS: The rats were treated with AlP (10 mg/kg; 5.55 x LD50; ig) and the survival time was noted. The effect of atropine (1 mg/kg, ip) and PAM (5 mg/kg, ip) was noted on the above. Atropine and PAM were administered 5 min after AlP. Plasma cholinesterase levels were measured spectrophotometrically in the control and AlP treated rats 30 min after administration. RESULTS: Treatment with atropine and PAM increased the survival time by 2.5 fold (1.4 h+/-0.3 h vs 3.4 h+/-2.5 h, P < 0.01) in 9 out of 15 animals and resulted in total survival of the 6 remaining animals. Plasma cholinesterase levels were inhibited by 47 %, (438+/-74) U/L in AlP treated rats as compared to control (840+/-90) U/L (P < 0.01). CONCLUSION: This preliminary study concludes that AlP poisoning causes cholinesterase inhibition and responds to treatment with atropine and PAM. PMID- 11730560 TI - Inhibitory effect of nomegestrol acetate on steroidogenesis of cultured granulosa cells from rat ovary in vitro. AB - AIM: To study the effect of nomegestrol acetate, a new synthetic progesterone on granulosa cells' viability and steroidogenesis function. METHODS: Granulosa cells were cultured in McCoy's 5A medium. Trypan blue stain was used to measure viable cells. FSH and testosterone were added to stimulate the steroid secretion. Specific RIA assay was used to evaluate the estrogen and progesterone secretion respectively. RESULTS: IC50 of nomegestrol acetate to damage cells is 6.85 mg/L (95 % confidence limits 5.36-8.75 mg/L). Nomegestrol acetate 0.45, 0.9, and 1.8 mg/L greatly inhibited the estrogen secretion from granulosa cells by 7.6 %, 12.5 %, 28.3 % in the presence of testosterone 0.5 micromol/L and FSH 10 U/L without affecting the number of viable cells. The secretion of progesteron were markedly decreased by 44.5 %, 53.3 %, and 62.0 % concurrently. CONCLUSION: Nomegestrol acetate directly inhibited the steroidogenesis of granulosa cells. PMID- 11730561 TI - Development of quantitative structure activity relationships in toxicity prediction of complex mixtures. AB - AIM: To predict the toxicity of mixtures of halogenated benzenes (narcotics). METHODS: Acute toxicity test of single chemicals and mixtures was performed using Photobacterium phosphoreum. Partition coefficients of mixtures were calculated by a special equation. Quantitative structure-activity relationship (QSAR) studies were carried out based on results of toxicity tests, n-octanol/water partition coefficient (Kow), and partition coefficient of mixtures (Kmix). RESULTS: 1) There was a good relationship between toxicity and lgKow for single compounds. 2) QSAR analysis showed a perfect correlation between the calculated Kmix and the mixture toxicity for binary mixtures. 3) Using the QSAR model of binary mixtures, the toxicity of other related mixtures containing different composition and fraction was predicted very well. CONCLUSION: The toxicity of halogenated benzene mixtures (narcotics) was predicted by QSAR. This kind of study is helpful for assessing the toxicity of narcotic mixtures. PMID- 11730562 TI - A 3D structural model of memapsin 2 protease generated from theoretical study. AB - AIM: To build a 3D structural model of memapsin 2 (M2) protease for theoretical study and drug design. METHODS: Structural alignment was performed based on multiple and pairwise sequence alignment of three templates. After the initial model was generated, energy minimization was completed by applying molecular mechanics method. Molecular dynamics (MD) technique was used to do further structural optimization. RESULTS: The 3D structural model of memapsin 2 was constructed. The model is reasonable according to several validation criteria. The active-site motifs of M2 are structurally supported by a beta-sheet rich domain and linked together with this domain through alpha helices. Tyr132 contained in beta-hairpin is a general characteristic of aspartic protease. The Calpha atom superimposing result is a direct verification that M2 is structurally unique but still belongs to the aspartic protease superfamily. CONCLUSION: The 3D structure model from our study is informative to guide future molecular biology study about M2 and drug design based on database searching. PMID- 11730563 TI - Uptake of cyclosporine A loaded colloidal drug carriers by mouse peritoneal macrophages in vitro. AB - AIM: To investigate the uptake of cyclosporine A loaded colloidal drug carriers by mouse peritoneal macrophage (MPM) in vitro. METHODS: The [3H]cyclosporine A loaded colloidal particles: polylactic acid nanospheres, polylactic acid nanocapsules, and microemulsions were prepared. The [3H]cyclosporine A loaded colloidal particles were incubated with MPM for 30 min at 37 degrees C, then the cells were separated from the colloidal particles and the radioactivity was measured by a liquid scintillation counter. RESULTS: In comparison to the cyclosporine A solution, the binding to polylactic acid nanospheres produced a 20 fold increase in the uptake of cyclosporine A by MPM in 30 min incubation, whereas some obvious decrease in the uptake of cyclosporine A by MPM was observed in the binding of cyclosporine A with polylactic acid nanocapsules or microemulsions. The surfactant coating and plasma protein adsorption were found to have marked effects on the uptake of cyclosporine A loaded nanospheres by MPM. CONCLUSION: Our present study indicated that colloidal drug carriers might affect the targeting of cyclosporine A to mononuclear phagocyte system. PMID- 11730564 TI - Effects of calcitriol and its analogues on interaction of MCP-1 and monocyte derived dendritic cells in vitro. AB - AIM: To study the effect of calcitriol [1,25(OH)2D3] and its analogues on the interaction of monocyte chemoattractant protein-1 (MCP-1) and in vitro generated monocyte-derived dendritic cells (MoDC). METHODS: MoDC were obtained by differentiating monocytes in exposure to GM-CSF and IL-4 for 5 d. mRNA expression of MCP-1 and its receptors were analyzed by RT-PCR, and protein production of MCP 1 by ELISA and migratory ability of MoDC in response to MCP-1 by a micromultiwell chemotaxis chamber assay. RESULTS: MoDC can express MCP-1 mRNA, and secret a low level of MCP-1 protein and has the ability to migrate to MCP-1 in corresponding to its expression of MCP-1 receptors. Calcitriol and its analogues with the same affinity to vitamin D receptor up-regulated the gene expression of both MCP-1 and its receptors, enhanced MCP-1 protein production and promoted the migratory ability of MoDC to MCP-1. CONCLUSION: The interaction of DC and MCP-1 found in this study may suggest a possible auto-regulatory role between DC and MCP-1 and the modulatory effect of calcitriol and its analogues on DC and MCP-1 might provide an understanding of their positive role in tumors. PMID- 11730565 TI - Gene expression of monocyte chemoattractant protein-1 in human monocytes by exposure to advanced glycosylation end products. AB - AIM: To explore the effects of advanced glycosylation end products (AGEP) on monocyte chemoattractant protein-1 (MCP-1) gene expression in human peripheral blood monocytes/macrophages (PBMC). METHODS: Expression of MCP-1 mRNA in PBMC incubated with AGEP-bovine serum albumin (AGEP-BSA) was examined by semi quantitative reverse transcription-polymerase chain reaction (RT-PCR) with beta actin as an internal standard. Sequencing of RT-PCR products was performed to confirm the specificity of amplification for MCP-1 gene. RESULTS: AGEP-BSA stimulated monocytes to express MCP-1 mRNA in a glucose-concentration-related fashion. The levels of MCP-1 mRNA were increased slightly when monocytes were exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 20 mmol/L), and increased markedly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 50 mmol/L), but decreased slightly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 80 mmol/L). Expression of MCP-1 mRNA was undetectable in freshly isolated monocytes, but was induced at 12 h and reached a maximal level at 24 h and was almost undetectable at 36 h after the monocytes were incubated with AGEP-BSA 200 mg/L (P < 0.01). CONCLUSION: AGEP enhanced MCP-1 mRNA expression in human PBMC. PMID- 11730566 TI - Prognostic value of dobutamine stress test in patients with septic shock. AB - AIM: To evaluate the prognostic effect of dobutamine stress test in patients with septic shock. METHODS: Patients with septic shock received intravenous infusion of dobutamine at 10 microg . kg-1 . min-1 for 1 h. Hemodynamics and oxygen metabolism was observed. A patient who was able to increase oxygen consumption index (VO2) by >15 % was designated as a responder to the test. RESULTS: In 47 patients with septic shock, twenty one responders and twenty six nonresponders were identified, and mortality was 33.3 % and 76.9 % respectively. After the dobutamine infusion, the responders showed increases in cardiac index (18.1 %), oxygen delivery index (12.7 %), VO2 (38.6 %), and oxygen extraction ratio (18.0 %), and reductions in systemic vascular resistance index (18.5 %). The nonresponders demonstrated increases in cardiac index (11.5 %), but no change in oxygen delivery, VO2, and oxygen extraction ratio. CONCLUSION: Dobutamine stress test might be a good predictor of outcome in patients with septic shock. PMID- 11730567 TI - Relationship of symptomatology, gender, and antipsychotic drug treatment with plasma homovanillic acid in schizophrenia. AB - AIM: To study the role of dopamine neurotransmission in schizophrenia and its drug treatment by assessing the relationship of plasma homovanillic acid (pHVA), a major central dopamine metabolite to various clinical parameters in schizophrenic patients. METHODS: pHVA was measured by high-performance liquid chromatography with electrochemical detection in a large cohort of both medicated and unmedicated DSM-IV schizophrenic patients. Prior to the measurement of pHVA, the patients were rated on the schedule for the assessment of positive and negative symptoms (PANSS). RESULTS: (1) pHVA in 46 patients receiving antipsychotic drugs was decreased, and in 58 drug-free patients increased, (7.4+/ 2.7) microg/L and (10+/-4) microg/L compared with a matched control group (9 microg/L+/-3 microg/L, n=62) (ANOVA F=8.57, df=2, P < 0.01), respectively. Within the drug-free group, pHVA was higher in the patients with a more negative symptom profile. (2) No significant correlation of pHVA with overall SAPS or SANS scores was apparent in the drug-free patients, although within the SANS subscales, a significant relationships to anhedonia-asociality (r=0.32, P < 0.05) was apparent. The male drug-free patients showed a positive correlation of pHVA with negative symptoms (r=0.42, P < 0.05) while females showed no significant relationship with any PANSS subscales. CONCLUSION: The results suggest that an increased dopaminergic metabolism is apparent in (male) schizophrenic patients with predominantly negative symptoms, supporting reports that this change in neuronal activity may be related to the neuropathological abnormalities seen in the disease, which may differ between males and females. Such neuronal deficits of developmental origin may thus result in an elevation/disinhibition of central dopamine metabolism in schizophrenia. PMID- 11730568 TI - Effects of AP-V and bicuculline on somatostatin-positive neurons in hypothalamus of rats subjected to acute hypobaric hypoxia. AB - AIM: To investigate the effects of 2-amino-5-phosphonovalerate-pharmacology (AP V) and bicuculline on somatostatin (SST)-positive neurons in hypothalamus of rats subjected to acute hypobaric hypoxia. METHODS: SST-immunoreactivity (IR) and somatostatin mRNA (SS mRNA)-positive neurons were measured by immunohistochemistry and in situ hybridization methods. RESULTS: Compared with control rats, SST-IR and SS mRNA-positive neurons in hypothalamic periventricular nucleus (PeV), paraventricular nucleus (PVN), and arcuate nucleus (ARC) increased after acute hypobaric hypoxia for 6 h (P < 0.01), and these effects were markedly inhibited by AP-V (10 microg, icv), a highly selective N-methyl-D-aspartate (NMDA) receptor antagonist, whereas were strongly enhanced by bicuculline (1.5 mg/kg, ip), a gamma-aminobutyric acid (GABAA) receptor antagonist. CONCLUSION: SST possibly participates in acute hypoxic reaction in hypothalamus, furthermore, glutamate and GABA can affect somatostatin release and synthesis in hypothalamus through NMDA and GABAA receptors respectively. PMID- 11730569 TI - Effects of CYP2C19 genotype and CYP2C9 on fluoxetine N-demethylation in human liver microsomes. AB - AIM: The present study was designed to define the kinetic behavior of fluoxetine N-demethylation in human liver microsomes and to identify the isoforms of cytochrome P-450 (CYP) involved in this metabolic pathway. METHODS: The kinetics of Ne formation of norfluoxetine was determined in human liver microsomes from six genotyped CYP2C19 extensive metabolizers (EM). The correlation studies between the fluoxetine N-demethylase activity and various CYP enzyme activities were performed. Selective inhibitors or chemical probes of various cytochrome P 450 isoforms were also employed. RESULTS: The kinetics of norfluoxetine formation in all liver microsomes were fitted by a single-enzyme Michaelis-Menten equation (mean Km=32 micromol/L+/-7 micromol/L). Significant correlations were found between N-demethylation of fluoxetine at both 25 micromol/L and 100 micromol/L and 3-hydroxylation of tolbutamide at 250 micromol/L (r1=0.821, P1=0.001; r2=0.668, P2=0.013), respectively, and S-mephenytoin 4'-hydroxylase activity (r=0.717, P=0.006) at high substrate concentration of 100 micromol/L. S mephenytoin (SMP) (a CYP2C19 substrate) at high concentration and sulfaphenazole (SUL) (a selective inhibitor of CYP2C9) substantially inhibited norfluoxetine formation. The reaction was minimally inhibited by coincubation with chemical probe, inhibitor of CYP3A4 (triacetyloleandomycin, TAO). The inhibition of fluoxetine N-demethylation at high substrate concentration (100 micromol/L) was greater in PM livers than in EM livers (73 % vs 45 %, P < 0.01) when the microsomes were precoincubated with SUL plus TAO. CONCLUSION: Cytochrome P-450 CYP2C9 is likely to be a major CYP isoform catalyzing fluoxetine N-demethylation in human liver microsomes at a substrate concentration close to the therapeutic level, while polymorphic CYP2C19 may play a more important role in this metabolic pathway at high substrate concentration. PMID- 11730570 TI - Pharmacokinetics of enantiomers of trans-tramadol and its active metabolite, trans-O-demethyltramadol, in human subjects. AB - AIM: To study the stereoselectivity in pharmacokinetics of the enantiomers of trans-tramadol (trans-T) and its active metabolite, trans-O-demethyltramadol (M1) in human subjects. METHODS: Trans-T hydrochloride sustained-release tablets were taken orally by 12 healthy male volunteers. After a multiple dosage schedule, the serum concentrations of (+)-trans-T, (-)-trans-T, (+)-M1, and (-)-M1 were determined in serum by high performance capillary electrophoresis (HPCE). RESULTS: (+)-Trans-T, (-)-trans-T, (+)-M1 and (-)-M1 in human serum were separated by HPCE. The linear range was 2.5-320 microg/L for the enantiomers of trans-T, and 2.5-50 microg/L for the enantiomers of M1. For the enantiomers of trans-T and M1, the intra-day and inter-day RSD were less than 15 % and 20 %, and the relative recoveries were 94.3 %-106.2 % and 90.4 %-107.8 %, respectively; the limit of quantitation was 1.25 microg/L. The serum concentrations of the enantiomers of trans-T reached a steady state in 12 subjects on d 4 after the initial administration. The steady state serum concentrations of (+)-trans-T were higher than that of (-)-trans-T at every sampling points in the subjects. The differences were significant in the main pharmacokinetic parameters between (+) trans-T and (-)-trans-T except Tmax. The serum concentrations of (-)-M1 were higher than that of (+)-M1 in most subjects and at most sampling time points. There were significant differences in Cmax and Cmin between the enantiomers of M1. CONCLUSION: The pharmacokinetics of trans-T and M1 was found to be stereoselective. (+)-Trans-T was shown to be absorbed completely, but eliminated more slowly. The pharmacokinetic stereoselectivity of M1 was different among human subjects. PMID- 11730571 TI - [Psicosocial rehabilitation in schizophrenia: variables related with rehabilitation]. AB - INTRODUCTION: The interest of vocational rehabilitation and integration in patients with chronic mental illness has grown in the last few decades. Different studies has tried identify predictors of vocational outcome with controversial results. In spite of the growing development of this field, there's a lack of studies in Spain looking for specific predictor variables of vocational outcome in schizophrenia. The aim of this study is to identify variables associated to vocational outcome in a social-vocational rehabilitation program (INICIA). METHODS: Two groups of subjects participating in a social-vocational rehabilitation program were selected based on different vocational outcomes. Sociodemographics, clinic, cognitive variables, social competence, basal work skills, and family variables, all assessed at intake, were compared between both groups. RESULTS: In our study, good initial scores in memory, comprehension, social and vocational skills, as well as, lower basal scores in tests of general psychopathology and psychotic symptomatology, were associated with a better vocational outcome. Patients with good insight and with lower number of previous admissions also showed better outcomes. CONCLUSIONS: Initial scores in different domains could be a useful guide to predict the outcome in some of the current rehabilitation programs. We come to the conclusion about the importance of developing future research studies about predictors of vocational outcome in patients with severe mental illness in our country. PMID- 11730572 TI - [Development, validity and reliability of a general addiction scale. A preliminary study]. AB - INTRODUCTION: The authors develop a General Scale to measure the intensity of the addiction to substances (not alcohol, not opiates) and addictive behaviors. METHODS: The General Scale is a self-scale compound by eleven items that was delivered to fifty and five students of the courses 5 masculine and 6 masculine of the Medicine of the University of Alcala (Madrid, Spain), and was them requested that applied said scale from different supposed addictive: tobacco; tea, coffee or cola drinks; chocolate; others sweet; alcohol; sex; use of the personal computer and/or Internet and/or videoplay; and to practice sports. Of that manner, each subject provided a total of 440 complimented scales.Finally, it was requested the subjects that indicated in a scale apart, their degree of addiction from the different exposed concepts. Those data served of external criterion. RESULTS AND DISCUSSION: The Scale is monodimensional, and shows a high construct validity (account 63% of the total variance obtained by a Factorial Analysis), a high alpha reliability (alpha: 0.94) and a good internal consistency (split-half method with the Spearman-Brown correction; R: 0.92). All items share with the general addiction concept that represents the total score of the Scale, an common variance proportion equal or over the 52%. CONCLUSION: The Scale seems be a valid and reliable instrument to compare groups of the calls new addictions of a measurable manner. PMID- 11730574 TI - [Psychopathology of bulimia nervosa: a multidimensional model]. AB - INTRODUCTION: In recent years, a number of studies have been carried out with the intention of isolating clinical dimensions in the psychopathology of bulimia nervosa. Although borderline personality has been considered a core element of the bulimic psychopathology by most of the authors, it has not been incorporated into any of these models. In this context, the present study was aimed at testing the consistence of the more complex model proposed until now, including in the analysis borderline personality as a clinical variable. SAMPLE AND METHODS: A group of 66 female patients fulfilling DSM-IV criteria for bulimia nervosa were assessed using a set of clinical instruments. The isolated items were processed using factor analysis techniques. RESULTS: Five basic dimensions of bulimia nervosa were obtained: 1. body disatisfaction; 2. restrictive eating behaviors; 3. purging behaviors; 4. emotional instability; and 5. disocial behavior. CONCLUSIONS: Our results support the idea that bulimia nervosa is a multidimensional condition. In our model, the dimension emotional instability incorporated borderline features, which tended to be strongly associated to self defeating behaviors and depressive symptoms. PMID- 11730575 TI - [Risperidone in the treatment of psychotic patients with opiate abuse and dependence]. AB - INTRODUCTION: 180 psychotic patients with opiate dependence and abuse (ICD-10) were included in an open label study. The study objectives were to evaluate safety and efficacy of risperidone for a six month follow-up period. The total mean dose was 4.4 (SD: 2.4 mg/daily; range: 0.5-12 mg/daily). METHODS: BPRS, CGI and DDS-SV were used to assess efficacy and UKU subscale for neurological side effects and spontaneous reports for safety. RESULTS: Risperidone treatment improved symptoms, disability of the included patients with a significant reduction in the mean total scores of BPRS, CGI and DDS-SV observed from the first month of treatment onwards. Risperidone also reduced illegal opiate abuse patients from 39% basedate to 18% at month 6. There was a significant reduction (p< 0.0001) in the total UKU subscale for neurological side effects scores from visit 1 onwards for studied sample. Risperidone was well tolerated by the study patients. From 165 elegible patients, just 10 (6.1%) discontinued treatment due to adverse reactions, 94% of the patients did not suffer any adverse event; the most frequent adverse events according spontaneous reports were extrapyramidal effects (3%) and anxiety (1.8%). DISCUSSION: Risperidone improved disability, psychotic symptoms and tolerability of these patients. Those results could mean an outstanding breakthrough in the treatment of these type of disorders and, if it is confirmed that risperidone can lead to abstinence, we would be before a new line of treatment for dual pathology. PMID- 11730576 TI - [Efficacy of gabapentin in a sample of bipolar patients]. AB - INTRODUCTION: Field studies support the idea of gabapentin being helpful in the maintenance, and in the treatment of some symptoms in bipolar patients. Our study evaluates relapse rates previous and after gabapentin administration in severe bipolar patients. METHODOLOGY: Gapabentine was administered to all consecutive bipolar patients from a relapse prevention program who could not be given lithium, valproic acid or carbamacepin because of his current medical condition or his past history of secondary effects or lack of response to those treatments. Number and severity of relapses were evaluated before and after gabapentin administration. RESULTS: Seven patients were included in the study. Medium maintenance period with gabapentine was 9 months. In the gabapentin period, relapses per month increased from 0.18 in the previous three years to 0.29. This may be due because six patients interrupted abruptly previous treatment in less than 1 week. Relapses severity, evaluated by measuring length and number of hospitalisations, and number of interviews by month, was similar to the three previous years, and better than the period from the beginning of the condition. Clinical Global Impression evaluated gabapentin as similar to previous mood stabilizers in five patients and better in two. Irritability and dysphoria improved in all the patients. CONCLUSION: Although gabapentin may be helpful in some patients, a clinical essay that shows its efficacy as add-on treatment is need. PMID- 11730577 TI - [Variability in attitudes and use conditions of electroconvulsive therapy. Results of a preliminary study]. AB - OBJECTIVE: Presenting the results of a pilot study on attitudes and use of ECT in Spain which was administrated to volunteers during a national congress of psychiatry. MATERIAL AND METHODS: A cross survey on a sample of psychiatrists (N= 125) which is not representative enough who attended the congress. A descriptive analysis of the outcomes is made. RESULTS AND CONCLUSIONS: There is a statistically significant association depending whether ECT is used in the work place or not. When it is used, there are 100% favorable attitudes. On the other hand, only 36% attitudes are favorable when ECT is not used in the work place. It is typically applied within a period of 8-10 treatment sessions, with a mean frequency of three times a week, performed by psychiatrists who treat with this technique less than 10 patients per year and using facilities within 1 to 5 years old. Maintenance ECT is unusual and it is only performed in monthly or fortnightly sessions. The commonest anesthetic agents are Propofol and Thiopental but this is unknown by 27.6% of the practitioners. The outcomes are contrasted with the consolidated guidelines on clinical practice of ECT. PMID- 11730578 TI - [Vocational definition in the medical students: The influence of personality]. AB - The vocational indefinition has been related by different authors with higher levels of personal immaturity, dependency sentiments or anxiety. In this paper, we analyses the possible rapport between specific personality traits and the vocational definition in the Medical Students. MATERIAL AND METHOD: It has been analyses 1484 students of 2nd year of Medicine in the Valencia University. They have responded an anonymous interview about their characteristic sociodemographics and their vocational interests. We determinate their Personality Profile with the 16 P.F. of Cattell. The done got has been analysed with Techniques of Regression Logistic. RESULTS AND CONCLUSIONS: The majority of the analysed population were female (62,7%) and with a medium age of 20 years. The majority of the students have definite vocationally in the 2nd year of Medicine: the 81% against the 19%. The students who haven't defined vocationally trend to be more unstable (C-), expedient (G-), strong minded (I-) and shrewd (N+). CONCLUSIONS: The results of our study seems to confirm the association between the career indecision in the Medical Students and specifics traits of personality, particularly the traits associated with the immaturity personal. PMID- 11730579 TI - [The epidemiology of eating disorders and the influence of mass media: a literature review]. AB - The aim of this study was to provide an update of the epidemiology of eating disorders (anorexia nervosa and bulimia nervosa), focusing on incidence and prevalence as well as other factors as age and sex. We made a review of the existing literature in the matter in the last decades. Likewise, we made an extent evaluation of the hypothetical pathogenic impact that exerts mass media (television and printed mass media) in these pathologies. We found an evident increase in incidence as well as prevalence of these disorders though it's far from being considered epidemic. We also found an undeniable influence of mass media in the genesis and maintenance of these disorders. However, mass media would exert a positive influence in the popularization of these entities to the general population that would result in a higher demand in general practice. PMID- 11730580 TI - [Swallowing phobia: a case report]. AB - Choking phobia is characterized by fear and avoidance of swallowing food, fluids or pills. It often appears in women after an episode of choking on food. We present the case of an adolescent beginning her phobia after a mononucleosis with severe dysphagia. Patient's solid restriction raised concerns about differential diagnosis with eating disorders. She was treated with psychoeducation, cognitive restructuring and graduated exposure in vivo, achieving a total recovery. PMID- 11730581 TI - [A clinical case of galactorrhoea after citalopram treatment]. PMID- 11730586 TI - [Diagnostic approach and treatment of inherited mitochondrial fatty acid oxidation disorders]. AB - Inherited mitochondrial fatty acid oxidation disorders are a complex set of genetically-based diseases in which up to 22 different entities are currently recognized. Their incidence is probably underestimated because a high level of diagnostic suspicion is required for their detection. Their clinical spectrum and prognosis are variable. In recent years knowledge of these diseases and improved treatment have reduced associated mortality. A common characteristic of all these diseases is hypoketotic hypoglycemia, although this is not constant and does not appear in the short-chain disorders and, sometimes, does not even appear in the medium-chain disorders. Cardiac or skeletal myopathy combined and/or hepatic involvement at periods of metabolic decompensation are typical, since these tissues depend on fatty acid oxidation. Diagnosis has been simplified by the study of acylcarnitines in blood, even in periods of metabolic stability. Determination of acylglycines, organic acids, carnitines, free fatty acids and 3 hydroxy-fatty acids, together with enzymic and genetic studies, complete the diagnosis. In certain circumstances, a provocation test should be carried out. Treatment basically consists of avoiding fasting, restricting fatty acid uptake and increasing carbohydrate uptake, depending on the type of metabolic disorder. Pharmacological treatment may also be added (carnitine, riboflavine or carbamylglutamate). PMID- 11730587 TI - [Medication errors in a neonatal unit]. AB - INTRODUCTION: Medication errors occur as a result of human mistakes or system flaws and could be prevented by studying and modifying the conditions that predispose to errors. In recent years, interest in medical errors has increased because of their health and economic implications. OBJECTIVES: To evaluate the frequency and types of prescribing errors in the Neonatology Unit of the Hospital 12 Octubre before and after an intervention to raise awareness of errors among the medical staff by comparing the frequency of error before and after the intervention. PATIENTS AND METHODS: We conducted a prospective pilot study in two phases. In the first phase, we studied prescribing errors by reviewing 100 prescriptions for newborns admitted to the Intensive or Intermediate Care Units. When the prescriptions were written, the neonatologists were unaware that the study would be performed. Legibility, dose, units used to express medications, route of administration, use of abbreviations, specification of dosage per kilogram of body weight and use of brand names were evaluated. The information was analyzed and an information-training intervention was performed in which the results were made known in a clinical session and recommendations for improving prescriptions were made. In the second phase, another 100 prescriptions were reviewed. The results were compared with those obtained before the intervention using the chi-squared test. RESULTS: In the first phase, 22 % of prescriptions were illegible or doubtful, 4 % contained dose errors and 28 % did not specify the route of administration. After the intervention, 8 % (p 0.005) of prescriptions were illegible, 4 % contained dose errors and 5 % (p 0.0001) did not specify the route of administration. Regarding other quality markers, the percentage of prescriptions specifying dosage per kilogram of body weight increased from 46 % to 78 %. Brand names were used in 21 %. Units were always expressed in abbreviations. All errors were severity index 0 or 1. CONCLUSIONS: The first step in prevention is recognition of mistakes. Increasing awareness among the medical staff of the consequences of errors improved the quality of prescriptions in our department. Awareness of the frequency and type of errors is the first step towards implementing strategies to reduce iatrogeny. PMID- 11730588 TI - [Usefulness of a software package to reduce medication errors in neonatal care]. AB - BACKGROUND: Many treatment errors in neonatal intensive care units are caused by the need to carry out a sequence of calculations to determine the dose and dilution of the drugs used. OBJECTIVES: To help in this task, we designed a spreadsheet (Neodosis) that helps clinicians and nurses to calculate the doses and standardize the dilutions of some of the drugs most commonly used in resuscitation and neonatal intensive care units. The aim of this study was to verify the usefulness and reliability of this software package. METHODS: A randomized, cross-over, controlled trial was conducted through simulated clinical cases in which the number of errors in the prescription data and the amount of time spent in making calculations, with and without the program, were evaluated. Fifty-four tests were performed by pediatricians, third- and fourth-year pediatric residents, and nurses. RESULTS: Without computer support, all three groups made errors (residents, pediatricians and nurses in descending order). When Neodosis was used, all the medical staff made significantly fewer errors. The greatest reduction was found in errors made by pediatric residents: minor errors decreased from 16 % to 2 % and major errors from 1.6 % to zero. When using the spreadsheet, the time spent by all groups in making the calculations was reduced by between one-third and one-half. CONCLUSIONS: The tests performed with simulated clinical cases revealed that the number of errors made by the healthcare personnel who participated in this study was not inconsiderable. The use of Neodosis helped physicians and nurses to make markedly fewer errors and also saved them time. PMID- 11730589 TI - [Balloon pulmonary valvuloplasty in a 1,680 g preterm infant]. AB - We report a preterm infant weighing 1,680-g with critical pulmonary stenosis. Pulmonary balloon valvuloplasty was successfully performed using a = 1.15 balloon/annulus ratio. PMID- 11730590 TI - [Cervical mass in an 8-year old boy]. PMID- 11730592 TI - [Recurrent respiratory papillomatosis: a cause of progressive respiratory distress]. AB - A 5-year-old girl was brought to the emergency department because of respiratory distress of 3 months' evolution that had worsened over the previous 3 weeks. Cervical radiograph and cervical computerized axial tomography showed an endolaryngeal mass almost completely occluding the airway. Urgent laryngoscopy revealed an endolaryngeal fleshy mass, which was excised during the procedure. Histological diagnosis was laryngeal papilloma. Although infrequent in children, laryngeal papilloma is the most common laryngeal tumor in pediatrics. It is currently named juvenile onset recurrent respiratory papillomatosis. The human papilloma virus (HPV) causes the disease by vertical transmission from a mother with active or latent vulvar HPV infection. PMID- 11730591 TI - [Achondrogenesis type II-hypochondrogenesis: radiological features.Case report]. AB - We present a case of lethal dysplasia in the neonatal period. The abnormality was suspected after ultrasonography of a pregnant woman presenting weak fetal movements revealed shortening of the extremities, voluminous cranium and polyhydramnios. Clinical and radiological findings showed platyspondylic dwarfism with short extremities, narrow thorax and hydropic appearance. The infant died on the third day of life from progressive respiratory distress. In the absence of histological, chondro-osseus and molecular studies, detailed clinical and radiological studies, as well as the lethal evolution during the neonatal period, guided the diagnosis of hypochondrogenesis. This entity, together with achondrogenesis II (and other dysplasias), forms part of the same spectrum of collagen type II abnormalities produced by a defect in the gene (COL2A1) that codifies collagen II, located in chromosome 12 I(12q13.1-13.2). When a heterozygote is produced, transmission is dominant autosomal. The phenotype shows wide variation and severity depends on the mechanism and location of the mutation. The definitive diagnosis is given by cytomolecular studies, while individualization of the different entities is based on histological data from the cartilage; clinical findings and skeletal radiology serve as a guide. PMID- 11730593 TI - [Precocious puberty and pituitary incidentaloma]. AB - The case of an 9-year-old girl with precocious puberty and a non-functioning pituitary adenoma is described. A review of the literature on the incidence, evolution and therapeutic options of pituitary incidentalomas in children and adults is performed. PMID- 11730594 TI - [Portal vein thrombosis. Review of the disease and report of two cases]. AB - Occlusion of the portal vein in patients with normal liver is a frequent cause of hypertension in children. The natural course of the disease is characterized by recurrent bleeding during the first years of life and the development of splenomegaly and hypersplenism. Because the risk of bleeding in these patients can decrease with age, non-aggressive management is preferred whenever possible. We report two cases of portal vein thrombosis in adolescents with no previous esophageal variceal bleeding. The patients were treated conservatively and, at the present moment, remain symptom-free. PMID- 11730595 TI - [Focal nodular hyperplasia of the liver]. AB - Focal nodular hyperplasia (FNH) of the liver is an uncommon benign lesion that usually occurs in young or middle-aged women. Although it has been associated with oral contraceptive use, no evidence supports this hypothesis. Most patients with this lesion are asymptomatic. We report the case of a 10-year-old boy with a history of transient epigastric abdominal pain over several days. Physical examination revealed a tender mass in the upper left quadrant. Abdominal ultrasonography revealed an 85 3 45 mm mass located in the left lobe of the liver. Computed tomography-guided biopsy showed histological changes consistent with FNH. Left hepatectomy was performed and the diagnosis was confirmed by histological study. FNH of the liver is extremely rare in children, especially in boys. Unlike adenoma, malignant transformation of FNH has not been described and consequently patients can be periodically followed-up with imaging tests, without need of surgery. The differential diagnosis includes other benign hepatic lesions such as hemangioma and hepatocellular adenoma. Surgical resection is indicated in symptomatic patients, usually with abdominal pain, cholestasis or palpable abdominal mass, and when the differential diagnosis, especially with hepatic adenoma, is unclear. PMID- 11730596 TI - [Total mobilization of the urogenital sinus in the treatment of cloaca]. AB - Persistent cloaca is one of the most complex forms of anorectal malformation in girls. Surgical treatment is complicated and laborious especially when the common channel is long. The aim of this report was to emphasize the importance of the joint mobilization of the urogenital sinus after separation from the rectum. This maneuver avoids separation of the urinary tract from the genital tract and reduces operating time by more than 60 %. This is only possible when the common channel of the cloaca is less than 3 cm. We present the cases of two girls treated in the last 2 years with this technique. Operating time was significantly shortened to approximately 4 hours and the procedure was considerably simplified. The functional and cosmetic results were excellent. To obtain good results in the treatment of cloaca, a precise clinical and radiologist diagnosis must be made, surgical needs should be prioritized and the appropriate technique chosen. The best procedure for patients whose common channel is less than 3 cm is total mobilization of the urogenital sinus, subsequently sutured to the perineum, and placement the rectum within the anorectal sphincter complex. PMID- 11730597 TI - [Ureteral ectopia as a cause of urinary incontinence in girls]. AB - The prevalence of ureteral ectopia is low and it is usually associated with pyeloureteral duplication. In these cases, incontinence is frequent. We present the cases of five female patients treated in our hospital for urinary incontinence due to ureteral ectopia and duplicity. The ectopic ureter reached the uterus in three patients and the vagina in the remaining two. The treatment chosen was heminephroureterectomy due to the presence of hypofunctional or dysplastic segments. Incontinence disappeared after surgery. PMID- 11730598 TI - [The responsibility of standards committees]. PMID- 11730599 TI - [Recommendations on medication and breastfeeding]. PMID- 11730601 TI - [Central apnea, brainstem infarctions and Pneumococcus meningitis]. PMID- 11730602 TI - [Staphylococcus hominis pneumonia in an immunocompetent girl]. PMID- 11730603 TI - [Tobacco, alcohol and illegal drug consumption, sexual activity and associated infectious diseases in delinquent adolescents]. PMID- 11730604 TI - [Suction injuries in public swimming pools]. PMID- 11730605 TI - [Unilateral laterothoracic exanthem of childhood]. PMID- 11730606 TI - [Multinodular globular mastocytosis]. PMID- 11730609 TI - [Medication errors in pediatrics]. PMID- 11730610 TI - [Use of alternating antipyretics in the treatment of fever in Spain]. AB - OBJECTIVE: To assess the frequency of the use of alternating antipyretics among Spanish pediatricians and to analyze the factors that determine this practice. METHODS: An anonymous survey of practicing pediatricians was conducted. Data on the use of antipyretics were collected. Factors related to alternating antipyretics were assessed by using multivariate correlation analyses. RESULTS: Three hundred twenty-four questionnaires were reviewed. The mean age of participants was 46.4 18.6 years (29-68 years). Of the responding pediatricians, 22.5 % worked in hospitals, 87.5 % in health centres and 24.7 % in in private practice. A total of 68.8 % of pediatricians (95 % CI: 63.4-73.8 %) alternated antipyretics. The combination of ibuprofen and acetaminophen was recommended by 96.9 %. The use of alternating antipyretics was positively associated with male sex (OR: 0.535; 95 % CI: 0.296-0.967; p 0.038), with few years' experience (OR: 0.952; 95 % CI: 0.922-0.984, p 0.005), with considering ibuprofen the drug of choice (OR: 2.324; 95 % CI: 1.235-4.372; p 0.009) and with the doctor's recommendation that the antipyretic be readministered in the event of slight increases in temperature (OR: 0.529; 95 % CI: 0.310-0.901; p 0.019). CONCLUSIONS: Despite the lack of scientific evidence to justify the practice, the use of alternating antipyretics is common in the treatment of febrile children. Predictive factors of this practice among pediatricians are male sex, having relatively little experience, considering ibuprofen as the drug of choice and recommending the administration of new doses of antipyretic to control mild fever. PMID- 11730611 TI - [Bronchiolitis and persistent wheezing. Is eosinophilia a risk factor?]. AB - OBJECTIVES: To investigate the final outcome of infants presenting an episode of bronchiolitis and to analyze the risk factors for the development of persistent wheezing, with special attention paid to the presence of eosinophilia during the acute episode. PATIENTS AND METHODS: We studied all the children aged less than 2 years who required hospitalization for bronchiolitis between 1990 and 1993. Eosinophil values during the acute phase were collected. Clinical evolution and outcome were classified in three groups: short-term persistent wheezing (if resolved before the child reached the age of 5 years), long-term persistent wheezing (if the child remained symptomatic after the age of 5 years) and no wheezing. Other risk factors for asthma such as familial history, passive smoking and respiratory syncytial virus isolation were also studied. RESULTS: We analyzed 170 patients (100 boys and 70 girls) aged 7.15 6 0.78 years who suffered an episode of acute bronchiolitis at the age of 5.4 4.2 months. One hundred thirteen patients (66.5 %) developed short-term persistent wheezing and 60 (35.3 %) developed long-term persistent wheezing. Fifty-six patients presented no wheezing. The number of patients with eosinophils > 1 % was significantly different in the three groups of patients (p 5 0.029). A Eosinophil values of > 1 % was associated with short-term persistent wheezing (p 5 0.013). Mean eosinophil values were higher in patients who developed wheezing at some time during evolution (p 0.028). A familial history of asthma was associated with the development of long-term persistent wheezing (p 0.033). CONCLUSIONS: In the population studied, eosinophil values of > 1 % during an episode of acute bronchiolitis in infancy was associated with a higher risk of developing persistent wheezing in the first 5 years of life. A familial history of asthma was associated with a higher risk of developing long-term persistent wheezing. PMID- 11730612 TI - [Nutritional status in the pediatric population of a rural area on the Ecuadorian coast]. AB - OBJECTIVES: To determine the effects of a diet based on carbohydrates on the nutritional status of a pediatric population through a study performed in a rural area of the province of Los Rios (Ecuador). PATIENTS AND METHODS: We evaluated 871 Ecuadorian children from a rural environment in an ambulatory medical clinic created for the purposes of the study. Basic anthropometric measurements (weigh, height, head circumference and upper arm circumference) and clinical data from physical examination were recorded. The anthropometric measurements were compared with the growth charts used in Spanish children, which were taken as the standard reference measurements. Information on nutrition was obtained from interviews with the mothers. RESULTS: A total of 871 children were evaluated (53 % boys; 47 % girls). Mean age was 6.39 years (range: 0-14 years; mode 4). Comparison of the data obtained with the standard growth charts for the Spanish pediatric population revealed significant differences in body mass index in boys aged 5-14 years and in girls aged 3-12 years and highly significant differences in boys aged 6-13 years and in girls aged 3-12 years. Significant differences in middle upper arm circumference were found in boys and girls aged 1-14 years and highly significant differences were found in boys aged 1-12 years and in girls aged 2-12 years. A middle upper arm circumference of less than 14 cm (slight malnutrition) was found in 6.67 % of boys and in 9.15 % of girls; a middle upper arm circumference of less than 12.5 cm (serious malnutrition) was found in 1.82 % of boys and in 2.82 % of girls. Nutritional interviews revealed that the overall caloric intake was approximately 1,500 cal/ day. Eight percent of calories came from carbohydrates, 15 % from fats and 5 % from proteins. CONCLUSIONS: A poor diet affects the child population's nutritional status qualitatively but not quantitatively. Nutritional deficit favors the prevalence of childhood diseases that could be avoided by suitable feeding and hygiene. Because of the lack of economic and health resources, prevention is the fundamental weapon for maintaining health in underdeveloped countries. Childhood nutritional status is influenced by diet, which in turn is influenced by physical, climatic, cultural and political factors, and by stress, which is provoked by infections and parents' behavior. Nutritional status in children is a clear reflection of the socioeconomic status of a given population. PMID- 11730613 TI - [Improvement in the results of surgical resection of hepatocellular carcinoma]. AB - AIM: To present the results of hepatectomies performed for hepatocellular carcinoma in a specialist unit and to compare the results of an initial period (1987-1993) with those obtained in a second period (1995-2000) in which the indications were limited to Child class A patients without portal hypertension. During the second period technical improvements such as intermittent selective hilar clamping and greater hiliar restrictions on transfusions were introduced. PATIENTS AND METHODS: One hundred and ten hepatectomies were performed in 105 patients with hepatocellular carcinoma in our unit over a 12-year period. Eighty percent of the tumors occurred in cirrhotic livers, mainly caused by hepatitis C virus. In the second period, upper gastrointestinal endoscopy was systematically performed to study the presence of varices. Hemodynamics studies were optionally performed to rule out portal hypertension. RESULTS: In the second period larger tumors were resected, a greater number of major hepatectomies were performed due to the increased frequency of hepatocellular carcinoma in non-cirrhotic liver, and fewer patients underwent transfusion. Early mortality was reduced from 21% to 1.8% and mean survival significantly increased from 37 to 52 months. Actuarial survival increased from 64% to 91% at 1 year and from 23% to 52% at 5 years in the first and second periods, respectively. Disease-free survival also increased significantly from 53% and 84% at 1 year and 27% and 40% at 5 years in the first and second periods, respectively. Analysis of the results in cirrhotic patients also showed a statistically significant improvement in early mortality and survival. Multivariate analysis of prognostic factors for survival demonstrated that the absence of blood transfusion, patients who underwent resection in the second period and the presence of pseudocapsules were independent factors for increased survival. CONCLUSIONS: The results of liver resection for hepatocellular carcinoma improved significantly due to the reduction in early mortality produced by more rigorous patient selection and the introduction of technical improvements. PMID- 11730614 TI - [Diagnostic utility of hepatic hemodynamics study in hepatoportal sclerosis]. AB - AIM: To evaluate the diagnostic utility of catheterization of the suprahepatic veins in patients with suspected non-cirrhotic portal hypertension as well as to characterize the clinical features and evolution of this process. PATIENTS AND METHODS: Hepatic hemodynamics studies, transjugular biopsy and venography were performed in seven patients with suspected non-cirrhotic portal hypertension. In all patients the combination of the three procedures gave the diagnosis of hepatoportal sclerosis, which was subsequently confirmed by direct portography and percutaneous or laparoscopic liver biopsy. RESULTS: Three patients presented esophageal variceal bleeding at diagnosis. Only one patient required intrahepatic shunting due to refractory bleeding. None of the patients required surgical shunting or presented alterations in liver function. CONCLUSIONS: Hepatic hemodynamics study with transjugular biopsy and venography is an effective procedure in the diagnosis of hepatoportal sclerosis and in most cases invasive confirmatory tests are not required. The course of the disease is relatively benign if variceal bleeding is controlled. The treatment of choice in these patients is pharmacological and endoscopic. PMID- 11730615 TI - [Degree of association between serum levels and genotype in alpha-1-antitrypsin deficiency. Clinical usefulness]. AB - AIM: To determine the degree of association between serum alpha-1-antitrypsin levels and its phenotypes as well as its clinical expression. PATIENTS AND METHODS: The alpha-1-antitrypsin genotype was identified using polymerase chain reaction followed by restriction enzyme digest in 212 patients in whom serum alpha-1-antitrypsin determination had been requested. The reasons for the request, the existence of pulmonary or liver disease, clinical diagnoses and functional repercussions were analyzed. RESULTS: Two hundred and twelve patients were evaluated (68% males; mean age: 34 20 years). In 23 patients (10.8%) a deficiency variant was found (one or two M alleles were lacking) and in 8 patients (3.8%) the genotype was ZZ. All patients with MM genotype had alpha-1 antitrypsin levels of 75 mg/dl or higher while none of the patients with ZZ genotype had levels higher than 40 ml/dl. All the patients with ZZ genotype showed alterations: 3 had pulmonary emphysema, 1 had chronic obstructive pulmonary disease and 4 had hypertransaminasemia. One patient with pulmonary emphysema had severe respiratory insufficiency while in the remaining patients with respiratory problems, respiratory insufficiency was slight or moderate. None of the patients with hypertransaminasemia showed echographic signs of portal hypertension or clinical or laboratory signs of reduced liver function. CONCLUSIONS: There is a close association between alpha-1-antitrypsin levels and the different genotypes. Consequently, in basal conditions with serum alpha-1 antitrypsin levels higher than 75 mg/dl genotyping is not required. The functional repercussions of deficiency variants in young adults is slight. PMID- 11730616 TI - [Current status of endoscopic retrograde cholangiopancreatography. What is the effect of the introduction of magnetic resonance cholangiography?]. AB - AIM: To determine the current status of endoscopic retrograde cholangiopancreatography (ERCP) and the effects of the introduction of magnetic resonance cholangiography (MRC) on ERCP and to evaluate the diagnostic yield of MRC since its introduction in obstructive biliary disease. MATERIAL AND METHODS: We performed a retrospective analysis of the ERCP carried out between January 1998 and December 2000 and of the MRC performed for suspected obstructive biliary disease from May 1999 to December 2000. When both techniques were performed, the diagnostic yield of MRC was evaluated. RESULTS: We performed 927 ERCP. A total of 45.3% of the patients were men. Mean age was 69.2 14.6 years. Treatment was performed on 688 occasions (77%), mainly sphincterotomy (69.9%) and placement of polyethylene (21.8%) or self-expanding (9.9%) prostheses. The number of ERCP performed each year was 261, 330 and 336 in 1998, 1999 and 2000, respectively; of these 76.6%, 80% and 76.9% were therapeutic. MRC was performed in 63 patients with suspected biliary disease. In 27 of 59 patients (45.8%) MRC was sufficient for diagnosis, avoiding the need for ERCP. In the 35 patients in whom both techniques (MRC and ERCP) were performed, the diagnostic yield of MRC compared with that of ERCP was: sensitivity 100%, specificity 50%, positive predictive value 87.1%, negative predictive value 100% and overall value 89%. The cases responsible for the low specificity (all due to choledocholithiases) were analyzed and a significant time lapse was found between the performance of MRC and ERCP (range: 7-35 days) during which choledocholithiasis probably resolved spontaneously. CONCLUSIONS: a) Contrary to what could be expected, the number of ERCP seems to be increasing with a high percentage of endoscopic treatment; b) Because of its diagnostic sensitivity, MRC is the ideal technique for eliminating the need for diagnostic ERCP, thus allowing greater development of the therapeutic aspect of ERCP. PMID- 11730617 TI - [Gallstone Ileus: results of analysis of a series of 40 patients]. AB - Gallstone ileus is an infrequent surgical emergency that nevertheless represents a not insignificant percentage of occlusions of the small intestine without strangulation. We present a retrospective study of 21 years (1980-2000) in 40 patients. Mean age was 76 years and most of the patients (85%) were women. Mean age and associated disease increased throughout the study period. Cases of gallstone ileus decreased in the second and third 5-year periods and clearly increased in the fourth (1996-2000). Localization of gallstones was ileum in 25 patients, jejunum in 9, duodenum in 3, colon in 1 and feces in 2. Only 12 patients (30%) received a correct preoperative diagnosis. Treatment was surgical in 38 patients with 7.5% mortality and 57.5% morbidity. Enterolithotomy was the technique of choice. A high degree of suspicion for gallstone ileus and early treatment may be the factors required to improve prognosis. The increase in life expectancy and associated comorbidity, which contraindicate elective surgery for cholelithiasis, may be contributing towards an increase in the population prone to developing gallstone ileus. PMID- 11730618 TI - [Peutz-Jeghers syndrome associated with multiple epithelial tumors]. AB - The association between Peutz-Jeghers syndrome and multiple epithelial neoplasias is infrequent. Tumors of the gastrointestinal tract, genitals and breast are common in this syndrome and tumors of the kidney and biliary tract are exceptional. We present the case of a 68-year-old woman with Peutz-Jeghers syndrome who presented with right kidney carcinoma and numerous small, bilateral tumors of the sex cords with annular tubules of the ovary. Right nephrectomy, double annexectomy and total hysterectomy were performed. The patient died two years later following the development of rectal carcinoma diagnosed by sigmoidoscopy and biopsy and an episode of obstructive jaundice compatible with cholangiocarcinoma. PMID- 11730619 TI - [Chronic colonic pseudo-obstruction secondary to neuroleptics]. AB - Colonic pseudo-obstruction is characterized by non-mechanical chronic colonic dilatation. It is an infrequent entity that can be provoked by multiple causes, among them pharmacological. We present the case of a 74-year-old female psychiatric patient who presented abdominal bloating, diarrhea, intense electrolytic alterations and marked radiographic colonic dilatation after treatment with a neuroleptic (zuclopenthixol decanoate). Organic obstruction and other causes were ruled out and the final diagnosis was chronic colonic pseudo obstruction secondary to the use of neuroleptics. Cisapride (20 mg/8 h) produced a slight improvement in symptoms but colonic dilatation was permanent. PMID- 11730620 TI - [Spontaneous regression of hepatocellular carcinoma in a cirrhotic patient]. AB - Hepatocellular carcinoma mainly affects patients with liver disease and rarely develops in healthy liver. Cirrhosis greatly increases the risk of developing this malignancy and consequently these patients are candidates for close follow up and surveillance. The most commonly used screening programs are abdominal ultrasonography and serum alpha-fetoprotein determination. Without treatment, the prognosis of hepatocellular carcinoma is poor and mortality continues to be significant even in patients with small tumors detected during follow-up. We present the case of a 71-year-old male patient with alcoholic cirrhosis. The patient was diagnosed during follow-up of a single hepatocellular carcinoma of 4 cm in diameter. Although he refused all treatment, evolution was favorable, serum alpha-fetoprotein levels returned to normal and ultrasonographic images compatible with hepatocellular carcinoma disappeared. Three years after diagnosis, the patient was completely asymptomatic. PMID- 11730621 TI - [Subgenomic replications of hepatitis C virus (HCV): new possibilities for hepatitis C prophylaxis and treatment]. PMID- 11730622 TI - [Liver resection in the treatment of hepatocellular carcinoma]. PMID- 11730623 TI - [Surgical treatment of severe lower gastrointestinal hemorrhage in Crohn's disease]. PMID- 11730624 TI - [Celiac disease in adults]. PMID- 11730625 TI - [Replacement of gastrostomy catheter using Savary dilators]. PMID- 11730628 TI - [Body mass index (BMI) and percent fat mass. A BMI > 27.5 kg/m2 could be indicative of obesity in the Spanish population]. AB - BACKGROUND: The World Health Organization (WHO) defines obesity as a condition of excessive fat accumulation to the extent that health and well-being are affected. For population studies, the measurement of weight, corrected for height, is still the method of choice (BMI). In Caucasian populations, the BMI cut-off point for obesity (30 kg/m2) corresponds with a percent body fat (PFM) of over 25% in young adult males and 35% in young adult females. However, the relation between BMI and PFM is not uniform among populations. It is important to define in each population the threshold of BMI which corresponds to the definition of obesity. PATIENTS AND METHOD: BMI was calculated in 282 subjects. PFM was analyzed using Bio-electrical impedance. Blood pressure, serum glucose and insulin, and the fasting insulin resistance index were also determined in all subjects to analyze the metabolic impact. RESULTS: The PFM of 25% in males and 35% in females corresponded to a BMI of 27.5 and 27.4 kg/m2, respectively. The slope of the relationship between the degree of obesity and comorbilities (insulinemia, fasting insulin resistance index and blood pressure) was strengthened above 26 kg/m2 in men and 24 kg/m2 in women. CONCLUSIONS: BMI that corresponds to a PFM previously defined as obesity is lower in our population in comparison with other Caucasian populations. Our results confirm that is impossible to compare the prevalence of obesity among populations using exclusively the BMI. It will be important to define, using reference methods, whether these findings have physiological impact or not, and if this cut-off of BMI determines an increment in cardiovascular and overall mortality using epidemiological approaches. PMID- 11730629 TI - [The ISCA (Systematic Interview of Alcohol Consumption), a new instrument to detect risky drinking]. AB - BACKGROUND: The World Health Organisation Collaborative Project on Alcohol and Primary Health Care has stressed the need to develop standardised screening tools to enable early identification. The aim of this study was to develop a new systematic tool to register alcohol consumption and to validate its usefulness in order to detect risky drinking in primary health care settings. SUBJECTS AND METHOD: The Systematic Interview of Alcohol Consumption (ISCA) was administered together with the Alcohol Use Disorders Identification Test (AUDIT), which was used as main external criterium, to 255 patients who attended 5 primary health care centers. RESULTS: The correlation between both procedures was highly positive and significant (r = 0.831; p < 0.001). The cut-off scores (> 28 for men and > 17 for women) showed an ISCA sensitivity rank to detect risky drinking of 70-81% for men and 46-100% for women. The ISCA specificity ranks were 82-99% and 97-100%, respectively. CONCLUSIONS: The ISCA seems to be useful to detect risky drinking and it is easy to administer by primary health care professionals. ISCA and AUDIT can be used indistinctly and complementarily. PMID- 11730630 TI - [Importance of 5569G/A polymorphism in intron 4 of HFE gene in the diagnosis of hereditary hemochromatosis]. AB - BACKGROUND: The presence of the 5569A polymorphism may lead to misdiagnosis of patients susceptible of hereditary hemochromatosis (HH). For that reason, samples containing the Cys282Tyr mutation were revised and the frequency of this polymorphism in our environment was assessed. PATIENTS AND METHOD: Twenty samples were retested and 56 controls were included. The study was performed by PCR-RFLP. RESULTS: The diagnosis was confirmed in 8 cases susceptible of error. However, an amplification deficiency of normal alleles was detected in 2 heterozygous (17%). The allelic frequency of the 5569A polymorphism in the control population was 14.3%. CONCLUSIONS: Although misdiagnosis was not committed, we recommend changing to any primer that does not include the 5569G/A polymorphism in the study of HH. PMID- 11730631 TI - [Smoking-attributable deaths in Spain in 1998]. AB - BACKGROUND: Between 1993 and 1997, smoking prevalence remained stable in Spain yet age-adjusted death rates by smoking-related diseases decreased. Our study aimed to estimate the burden of smoking-attributable mortality in Spain in 1998. POPULATION AND METHOD: Spain's smoking prevalence, mortality and relative risks for death from the Cancer Prevention Study II were used to estimate smoking attributable mortality in the population aged 35 years and over. RESULTS: In 1998, 55,613 deaths were attributable to smoking. One out of 4 deaths in males and one out of 40 deaths in females were attributable to tobacco. Two thirds of the attributable mortality corresponded to deaths due to lung cancer, chronic obstructive pulmonary disease, ischemic heart disease and stroke. CONCLUSIONS: Smoking actually represents a remarkable burden of avoidable deaths in Spain. Smoking-attributable mortality appears to continue increasing in the last years. PMID- 11730632 TI - [Long life for the latest update on the Declaration of Helsinki?]. PMID- 11730633 TI - [Role of doctors as experts under the Spanish new civil judgment 1/2000 law]. PMID- 11730634 TI - [Depressive pseudodementia: clinico-therapeutical implications]. PMID- 11730635 TI - [Fever and headache in a 41-year-old male]. PMID- 11730636 TI - [Mitochondrial involvement in antiretroviral therapy-related lipodystrophy of HIV infected patients]. PMID- 11730637 TI - [Utilization of the questionnaire PrimeMD for the detection of mental disorders among Latin American and Spanish native speaker immigrant women]. PMID- 11730638 TI - [Increase of viral load of Epstein-Barr virus at diagnosis of primary brain lymphoma in a kidney transplanted patient]. PMID- 11730639 TI - [Diabetic mastopathy: source of confusion with breast carcinoma]. PMID- 11730640 TI - [Bupropion and smoking habit]. PMID- 11730642 TI - [The image of the week]. PMID- 11730643 TI - Optic nerve protection, regeneration, and repair in the 21st century: LVIII Edward Jackson Memorial lecture. AB - PURPOSE: To present the current status and clinical implications of optic nerve protection, repair, and regeneration after experimental injury in mammals, including nonhuman primates. DESIGN: Optic nerve and neuro-ophthalmology experimental study review. METHOD: Synthesis of experimental data regarding experimental studies of optic nerve protection, repair, and regeneration. RESULTS: Under certain conditions, mammalian retinal ganglion cells can be prevented from dying despite injury to the cell bodies or their axons, injured mammalian retinal ganglion cells whose axons have degenerated can be induced to extend new axons, and regenerating axons can reach their correct targets in the central nervous system. In addition, stem cells can be induced to become retinal ganglion cells. CONCLUSIONS: It may soon be possible to preserve and restore vision in persons whose sight is threatened or has been lost from disease or damage to the optic nerve. PMID- 11730645 TI - Persistent vertical binocular diplopia after cataract surgery. AB - PURPOSE: To report the incidence of, and factors associated with, persistent vertical diplopia after cataract surgery. DESIGN: Consecutive interventional case series. METHODS: Retrospectively, all adult patients examined during a five year, five month period because of new onset persistent (>3 months) vertical binocular diplopia after cataract surgery were analyzed. All patients had their cataract surgery at the same outpatient ophthalmic surgery center, and were referred to the author, enabling calculation of incidence. Trends in anesthesia type and strabismus complications therefrom were also assessed. Comparison was made between ophthalmologist-administered retrobulbar anesthesia versus anesthesia staff-administered retrobulbar anesthesia. Incidence during a period in which hyaluronidase was not incorporated in the retrobulbar anesthetic was calculated. RESULTS: Persistent vertical diplopia occurred after cataract surgery in 32 (0.18%) of 17,531 eyes that had cataract surgery. No patient whose cataract surgery was conducted with topical anesthesia (3817 eyes) had persistent vertical diplopia, whereas 32 (0.23%) of the 13714 eyes whose cataract surgery was done after retrobulbar anesthesia were affected. No cases of persistent postoperative diplopia were found among 7410 cataract surgery eyes after retrobulbar injection given by one cataract surgeon. There was a threefold greater number of left eyes involved than right eyes (P <.005). No significant (P >.20) increase in cases of persistent vertical diplopia was noted during a period of hyaluronidase shortage. CONCLUSIONS: In this study, persistent binocular vertical diplopia after cataract surgery occurred in 0.23% of cases in which retrobulbar anesthesia was performed. No cases were found after topical anesthesia. Occurrence may be technique related. PMID- 11730644 TI - Cluster of Mycobacterium chelonae keratitis cases following laser in-situ keratomileusis. AB - PURPOSE: To describe a cluster of Mycobacterium chelonae keratitis cases involving patients who underwent laser in-situ keratomileusis (LASIK) at a single refractive surgery center. DESIGN: Descriptive case series of four patients and cohort study to identify disease associations. METHODS: Examination schedules, diagnostic tests, and therapy were based on best medical judgment. Isolates from three patients were compared by pulsed-field gel electrophoresis. Epidemiologic studies were performed to identify the source of infection. RESULTS: Seven of eight eyes developed M. chelonae keratitis following bilateral simultaneous LASIK. Each patient was thought to have diffuse lamellar keratitis initially, but all seven eyes were noted to have opacities suggestive of infectious keratitis by 13 to 21 days after surgery. All eyes had undergone hyperopic LASIK over four days in April 2001 by one surgeon in a community-based refractive surgery center. A cohort study of all patients undergoing LASIK at the same center in April 2001 revealed that M. chelonae keratitis occurred only in persons undergoing correction of hyperopia (seven of 14 eyes vs. none of 217 eyes undergoing myopic LASIK, P <.001). The only difference identified between procedures was use of masks created from a soft contact lens in hyperopic LASIK. Three isolates (three patients) were indistinguishable by pulsed-field gel electrophoresis. Eyes were treated with a combination of antimicrobial agents, including topical azithromycin in three patients, with resolution of infection in all eyes over 6 to 14 weeks. The source of infection was not identified on environmental cultures. CONCLUSION: Postoperative nontuberculous mycobacterial keratitis can occur in an epidemic fashion following LASIK. Topical amikacin, azithromycin, clarithromycin, ciprofloxacin, or a combination of these agents, appears to be effective treatment for these infections. PMID- 11730646 TI - Agreement among clinicians in the recognition of patterns of optic disk damage in glaucoma. AB - PURPOSE: To determine the interobserver and intraobserver agreement in the recognition of different patterns of glaucomatous optic disk damage and evaluate if these patterns changed over time in patients followed for a number of years. METHODS: Patients with early to moderate glaucoma (n = 105) were consecutively enrolled to participate in a prospective observational study. In the first part of the present study, optic disk photographs obtained closest to patient's entry date in the prospective study were classified in a masked fashion by three observers according to the pattern of optic disk damage into one of the following: (1) focal, (2) myopic, (3) senile sclerotic, (4) concentric cup enlargement, (5) normal appearance, or (6) miscellaneous (those disks that did not qualify for any of the other groups). The observers were also asked to assign a confidence score for each classification, ranging from 1 (low confidence) to 5 (highest confidence). The three observers reclassified the photographs after a minimal period of 2 months, in order to assess intraobserver agreement. In a second part of the study, one observer reviewed, in a masked fashion, all the optic disk photographs that had been taken during the routine follow-up of the 105 patients in order to evaluate whether the classification of disk pattern changed over time. RESULTS: Intraobserver agreement yielded kappa values (95% confidence interval [CI]) from 0.51 (CI, 0.40 to 0.62) to 0.85 (CI, 0.77 to 0.93) depending on the observer. Interobserver agreement kappa values between all three observers was 0.40 (CI, 0.35 to 0.46), but it improved if photographs classified with moderate or higher degrees of confidence were included (0.52 [CI, 0.44 to 0.60]). During a mean follow-up period of 8.2 +/- 4.8 years, 41.6% of the eyes were always classified into the same group and 23.6% of the eyes were classified at least once into two or more of the four patterns of damage (groups 1 to 4). CONCLUSIONS: Intraobserver agreement on the pattern of optic disk damage was generally very good, with some variation among the observers. Interobserver agreement was reasonable and increased proportionally with the confidence in the classification. During long-term follow-up, the pattern of optic disk damage usually did not change. This type of classification can probably be used accurately in clinical practice. PMID- 11730647 TI - Scanning laser polarimetry in a selected group of patients with glaucoma and normal controls. AB - PURPOSE: To evaluate the ability of scanning laser polarimetry to discriminate between subjects with glaucoma with specific patterns of visual field defect and normal controls. METHODS: This cross-sectional, prospective study in a glaucoma practice, focused on subjects with glaucoma with predefined types of visual field defect, including advanced (group A, n = 14), localized (group L, n = 46), or mixed (diffuse and localized) defects (group M, n = 22) and normal controls (n = 32). Scanning laser polarimetry was performed in one study eye per subject. Two methods of analysis were used: a subjective analysis, in which examination printouts with the image of the optic disk manually blocked were classified by two observers masked to the diagnosis, and a logistic regression analysis of the retardation parameters included in the printouts. RESULTS: The observers correctly identified 97% of the controls and 68% of subjects with glaucoma (overall correct classification of 77%), with 93%, 70%, and 47% of patients from groups A, L, and M, respectively, being correctly identified. The best discrimination obtained with the logistic regression correctly identified 69% of controls and 94% of glaucoma subjects (overall correct classification of 87%). The performance was only slightly better for cases from group A compared with L and M. CONCLUSIONS: Subjective assessment of the scanning laser polarimetry standard printout of single eyes might not be sensitive enough to detect cases of glaucoma with localized or milder mixed types of visual field defect. The discriminating ability of scanning laser polarimetry improves slightly when logistic regression analysis is employed. PMID- 11730648 TI - Association of retinal vessel caliber and visual field defects in glaucoma. AB - PURPOSE: This study evaluates the asymmetry of peripapillary retinal vessel caliber between inferior and superior hemispheres in eyes with visual field defects predominantly in one hemifield. DESIGN: Observational case series. METHODS: In a retrospective study, 64 eyes of 64 patients with primary open-angle glaucoma who had a marked difference in visual field defects between hemifields and who had no history of diabetes, trauma, or vascular occlusive disease were studied. The diameters of the superior and inferotemporal vessels were measured at the optic disk border with calipers on an enlarged image. RESULTS: In 64 eyes, the average ratio of the superior temporal artery diameter to inferotemporal artery diameter was significantly greater in the eyes with predominantly superior visual field defects as compared with those with inferior defects (1.10 +/- 0.22 vs. 0.92 +/- 0.19, respectively, P =.002, two-tailed t test). This indicates that the arteriole corresponding to the hemifield with the greater visual field defect was narrower than the arteriole in the other hemifield. This relationship was confirmed using chi(2) analysis (P =.002) comparing the proportions of eyes with ratios greater or less than normal vessel caliber ratios (normal ratio = 0.95 from data reported by Jonas and associates to the location of the dominant field defect. No statistically significant relationship was detected between retinal vein diameter and localized visual field defects, as determined by both the unpaired t test and chi(2) analysis. CONCLUSION: In eyes with primary open-angle glaucoma, this study demonstrates a strong association between decreased peripapillary arteriole diameter and visual field defects in the corresponding hemifield. This reflects either an ischemic basis for glaucomatous damage or vascular constriction when there are fewer axons to nourish. PMID- 11730649 TI - Evaluation of travoprost as adjunctive therapy in patients with uncontrolled intraocular pressure while using timolol 0.5%. AB - PURPOSE: To evaluate the intraocular pressure-lowering efficacy and safety of travoprost 0.0015% and 0.004%, dosed daily in the evening compared with vehicle, in patients with open-angle glaucoma or ocular hypertension, whose intraocular pressure was not adequately controlled on timolol 0.5% twice daily (twice daily). METHODS: Subjects who qualified at screening began a run-in period dosing timolol twice daily for 3 weeks. If the subjects had an intraocular pressure of 24 to 36 mm Hg at 8 AM and 21 to 36 mm Hg at 10 AM and 4 pm in at least one eye on timolol, they were randomized to one of two concentrations of travoprost (0.0015% or 0.004%) or vehicle solution every day and were followed for 6 months. Four hundred twenty-six subjects were randomized. The mean intraocular pressure at 8 AM, 10 AM, and 4 PM in the patient's eye with the higher intraocular pressure was used for the analysis. RESULTS: Mean baseline values (25 mm Hg) for subjects at eligibility (while maintained on timolol) were not significantly different (P <.0001) among the treatment groups. The intraocular pressure was lowered an additional -5.7 to -7.2 mm Hg and -5.1 to -6.7 mm Hg in the travoprost 0.004% and 0.0015% concentrations, respectively. These changes were significantly (P < or =.0001) different from the vehicle group (-1.3 to -2.8 mm Hg). The intraocular pressure range on treatment at all visit times over the 6-month treatment period ranged from 17.9 to 19.2 mm Hg for travoprost 0.004% and 18.3 to 20.1 mm Hg for travoprost 0.0015% compared with 22.4 to 24.1 mm Hg for vehicle. Average hyperemia scores ranged from trace to mild (mean 0.5 on a scale of 0 = none/trace; 1= mild; 2 = moderate; 3 = severe) for all treatment groups. No iris pigmentation changes were observed in any patient during this study. There were no clinically or statistically significant changes from baseline in visual acuity, ocular cells and flare, fundus parameter, cup-to-disk ratio and visual field between the treatment groups. There were no serious adverse events reported for any treatment group. CONCLUSIONS: Travoprost produced clinically relevant and statistically significant additional intraocular pressure reductions from baseline when used adjunctively with timolol in subjects with open-angle glaucoma or ocular hypertension. PMID- 11730650 TI - Predictive value of a 1-day, 1-eye trial on the 1-month ocular hypotensive response of latanoprost 0.005%. AB - PURPOSE: To determine if a 1-day, 1-eye trial of latanoprost 0.005% was predictive of the 1-month intraocular pressure response in patients with uncontrolled open-angle glaucoma on maximally tolerated medical therapy without latanoprost. METHODS: One eye of 39 consecutive patients was enrolled in prospective trial of latanoprost 0.005%. An investigator masked to treatment eye and intraocular pressure result measured intraocular pressure at 1 day and 1 month after treatment. The diagnostic precision of the 1-day, 1-eye trial for the 1-month result was determined at a 10%, 20%, and 30% reduction from baseline intraocular pressure. RESULTS: Treatment produced a mean +/- SD (range) decrease in intraocular pressure from 20.9 +/- 4.6 (14 to 35) mm Hg to 16.6 +/- 3.6 (9 to 26) mm Hg at 24 hours (P <.0001, paired t test). This reduction in intraocular pressure was similar at 1 month with mean +/- SD (range) intraocular pressure of 16.1 +/- 3.1 (9 to 22) mm Hg (P <.0001, paired t test). The accuracy of the 1 day, 1-eye trial for the 1-month response at a 10%, 20%, and 30% reduction of intraocular pressure from baseline intraocular pressure was 96% (chi square = 29.5, P <.001), 74% (chi square = 8.4, P =.004), and 80% (chi square = 10.2, P =.001), respectively. CONCLUSION: A 1-day, 1-eye trial of latanoprost 0.005% is predictive of the 1-month intraocular pressure response in patients with uncontrolled open-angle glaucoma. PMID- 11730651 TI - Acute effects of sldenafil ctrate (Viagra) on intraocular pressure in open-angle glaucoma. AB - PURPOSE: To assess the acute effects of sildenafil citrate (VIAGRA) on the intraocular pressure (IOP) of patients with chronic open-angle glaucoma. DESIGN: This was a double-blind, randomized, placebo-controlled, crossover study, in which 15 patients received a single oral dose of sildenafil 100 mg or matching placebo on two separate occasions. METHODS: Fifteen subjects aged 63 +/- 14 years (mean +/- SD) with bilateral chronic open-angle glaucoma were administered a single oral dose of sildenafil 100 mg or matching placebo on two separate occasions at least 3 days apart. IOP was measured in both eyes by Goldmann ap planation tonometry at baseline and then at 1-5 hours after dosing. Brachial artery systolic and diastolic blood pressures were determined by sphygmomanometry, and heart rate was also monitored at baseline and 1-5 hours after dosing. RESULTS: Compared with placebo, no statistically or clinically significant change in IOP was detected after a single dose of sildenafil 100 mg (P =.20). Moreover, no significant change in mean systemic blood pressure (P =.12) or heart rate (P =.72) was detected after treatment with sildenafil. CONCLUSION: At the maximum therapeutic dose of 100 mg, sildenafil did not produce any significant acute change in IOP in men with chronic open-angle glaucoma. This information is of importance for patients with glaucoma receiving sildenafil for treatment of erectile dysfunction. PMID- 11730652 TI - Risk factors for failure of 5-fluorouracil needling revision for failed conjunctival filtration blebs. AB - PURPOSE: To investigate the risk factors for failure of 5-fluorouracil (5-FU) needling revision, a useful procedure for restoring a failed filtration bleb. DESIGN: Interventional case series. METHODS: Retrospectively conducted study. SETTING: Institutional. STUDY POPULATION: Sixty-four eyes of 64 consecutive glaucoma patients that underwent 5-FU needling revisions for failed filtering bleb following either trabeculectomy or phaco-trabeculectomy with or without adjunctive mitomycin C (MMC). OBSERVATION PROCEDURES: Goldmann applanation tonometry, Kaplan-Meier survival analysis, and Cox proportional hazards regression analysis. MAIN OUTCOME MEASURES: Successful outcome of the initial 5 FU needling revision, arbitrarily defined as target intraocular pressure (IOP) control with not more than two topical glaucoma medications and no additional 5 FU needling or other surgical procedures, was analyzed by Kaplan-Meier survival analysis, and risk factors for failure of the initial 5-FU needling revision were analyzed by Cox proportional hazards regression analysis. RESULTS: The cumulative success rate of the initial 5-FU needling revision was 45% at 1 year, 33% at 2 years, and 28% at 4 years. Failure of the initial 5-FU revision correlated significantly with preneedling IOP > 30 mm Hg (P =.0003), lack of MMC use during the previous filtration surgery (P =.013), and IOP >10 mm Hg immediately following needling revision (P =.0012) according to Cox's proportional hazards regression analysis. CONCLUSIONS: Pre-needling IOP > 30 mm Hg, lack of MMC use during the previous filtration surgery, and IOP > 10 mm Hg immediately after needling were found to be significant risk factors for failure of the initial 5 FU needling procedure. Therefore, it is important to monitor IOP closely following needling revision in those patients with such risk factors. They are more likely to require additional therapeutic interventions, including repeat needling revisions. PMID- 11730653 TI - Duration of vitrectomy and postoperative cataract in the vitrectomy for macular hole study. AB - PURPOSE: To report the association between duration of vitrectomy, as well as other risk factors, and the progression of nuclear sclerosis and posterior subcapsular cataract in the Vitrectomy for Macular Hole Study. DESIGN: A cohort study nested within a randomized controlled clinical trial. METHODS: Using a system similar to the Lens Opacities Classification System II, nuclear sclerosis (NS) and posterior subcapsular cataract (PSC) were scored in the vitrectomy and fellow eye of 74 patients at baseline and at 6, 12, and 24 months postoperatively. Age, baseline blood pressure and refractive power, and duration of surgery were evaluated as risk factors for NS or PSC progression and cataract extraction. RESULTS: The incidence of NS progression in the surgical group of vitrectomy eyes was 81% at 6 months, 98% at 1 year, and 100% at 2 years of follow up. In contrast, NS progression in the control group of fellow eyes was only 18% at 6 months, 20% at 1 year, and 8% at 2 years. The incidence of PSC progression in the surgical group remained at approximately 11% throughout follow-up, which was not significantly higher than the 3% to 5% incidence in the control group. Vitrectomy was significantly related to progression of NS cataract (P <.001) and cataract extraction (P <.01). No statistically significant differences were found for NS scores, PSC scores, or progression rates between eyes that had less than median surgical duration (60 min.) or more than the median surgical duration. Additionally, no significant differences were found when eyes that experienced 45 minutes or less surgical duration were compared with eyes that endured more than 75 minutes surgical duration. Age, blood pressure, and refractive power were not found to be predictors for NS and PSC progression. CONCLUSIONS: Although vitrectomy is a risk factor for NS progression, the duration of vitrectomy does not increase the risk. PMID- 11730654 TI - Comparison of three techniques of foveal translocation in patients with subfoveal choroidal neovascularization resulting from age-related macular degeneration. AB - PURPOSE: To report the results of three methods of foveal translocation in the presence of subfoveal choroidal neovascular membrane resulting from age-related macular degeneration. METHODS: We treated 51 eyes of 51 consecutive patients with subfoveal choroidal neovascular membranes resulting from age-related macular degeneration with one of three techniques of foveal translocation surgery: foveal translocation with partial retinotomy (n = 6), limited translocation (n = 9), and translocation with 360-degree retinotomy (n = 36). All patients were followed for at least 6 months postoperatively. The size of the choroidal neovascular membrane and the amount of foveal displacement, the best-corrected visual acuity, and complications were recorded preoperatively and postoperatively. RESULTS: The mean distance of the foveal translocation was greater in the 360-degree retinotomy group (3340 microm) than in the partial retinotomy (1060 microm, P <.001) and the limited translocation groups (1120 microm, P <.001). A final visual acuity of 20/200 or better was achieved in two eyes (33%) in the partial retinotomy group, seven eyes (78%) in the limited translocation group, and 23 eyes (64%) in the 360 degree retinotomy group. The final visual acuity improved by 0.2 logarithm of minimal angle of resolution (logMAR) unit or more in one eye (17%), one eye (11%), and seven eyes (19%), respectively. The final visual acuity was maintained within 1 line in zero eyes, five eyes (56%), and 19 eyes (53%), respectively. A retinal detachment developed postoperatively in five eyes (83%), zero eyes (0%), and 15 eyes (42%), respectively. CONCLUSIONS: A significant number of patients improved or maintained best-corrected visual acuity after translocation with 360 degree retinotomy, and limited translocation, whereas translocation with 360 degree retinotomy is suitable for larger choroidal neovascular membranes because it resulted in the greatest foveal displacement among the three translocation procedures. PMID- 11730655 TI - Scotoma and fixation patterns using scanning laser ophthalmoscope microperimetry in patients with macular dystrophy. AB - PURPOSE: We used scanning laser ophthalmoscope microperimetry to evaluate the retinal scotoma and the fixation points in the patients with macular dystrophy. METHODS: We studied 10 eyes of five patients with macular dystrophy (three patients with cone dystrophy and two patients with Stargardt disease). The mean patient age was 37 years (range, 13 to 64 years). An estimation of scotoma and fixation points on the retina was performed using scanning laser ophthalmoscope microperimetry. RESULTS: All 10 eyes (100%) had one of two types of dense scotoma: type one was a dense ring scotoma (five eyes, 50%), and type two was a dense central scotoma (five eyes, 50%) that included the center of the fovea. In all eyes with a dense ring scotoma, the fixation points were stable and did not shift. In all eyes with a dense central scotoma, the fixation shifted. The logarithm of minimal angle of resolution of the visual acuity in the eyes with the dense central scotoma was significantly worse than that of eyes with the dense ring scotoma type (P =.005). CONCLUSIONS: Scanning laser ophthalmoscope microperimetry findings demonstrate two types of dense scotoma (dense ring scotoma and dense central scotoma) in the patients with macular dystrophy. The two types of dense scotoma affect the shifting of the fixation points and the stability of fixation and may result in the difference in visual acuity in the patients with macular dystrophy. PMID- 11730656 TI - Computerized method of visual acuity testing: adaptation of the amblyopia treatment study visual acuity testing protocol. AB - PURPOSE: To report a computerized method for determining visual acuity in children using the Amblyopia Treatment Study visual acuity testing protocol. METHODS: A computerized visual acuity tester was developed that uses a programmed handheld device that uses the Palm operating system (Palm, Inc, Santa Clara, California). The handheld device communicates with a personal computer running a Linux operating system and 17-inch monitor. At a test distance of 3 m, single letters can be displayed from 20/800 to 20/12. A C program on the handheld device runs the Amblyopia Treatment Study visual acuity testing protocol. Using this method, visual acuity was tested in both the right and left eyes, and then the testing was repeated in 156 children age 3 to 7 years at four clinical sites. RESULTS: Test-retest reliability was high (r =.92 and 0.95 for and right and left eyes, respectively), with 88% of right eye retests and 94% of left eye retests within 0.1 logarithm of minimal angle of resolution (logMAR) units of the initial test. The 95% confidence interval for an acuity score was calculated to be the score +/- 0.13 logMAR units. For a change between two acuity scores, the 95% confidence interval was the difference +/- 0.19 logMAR units. CONCLUSIONS: We have developed a computerized method for measurement of visual acuity. Automation of the Amblyopia Treatment Study visual acuity testing protocol is an effective method of testing visual acuity in children 3 to 7 years of age. PMID- 11730658 TI - Wavefront guided ablation. PMID- 11730657 TI - Renal-coloboma syndrome: report of a novel PAX2 gene mutation. AB - PURPOSE: To report a novel sporadic PAX2 gene mutation in a child with atypical bilateral optic nerve coloboma and congenital renal hypoplasia. DESIGN: Observational case report and experimental study. METHODS: Mutational analysis of the PAX2 gene in a family. RESULTS: A 9-year-old patient with a history of renal transplantation for congenital renal hypoplasia was found to have bilateral optic nerve coloboma during ophthalmic examination for cytomegalovirus retinitis. A previously unreported mutation in exon 2, delT 602 leading to a prematurely truncated protein was identified in the child but in neither of her parents, demonstrating a de novo mutation or germline mosaicism. CONCLUSIONS: The causal relationship between PAX2 gene mutations and renal-coloboma syndrome is further supported by this novel mutation. Awareness of the systemic associations with optic nerve abnormalities and the ocular findings in syndromic renal diseases will facilitate the management of these highly variable disorders. PMID- 11730659 TI - Iatrogenic keratectasia after a deep primary keratotomy during laser in situ keratomileusis. AB - PURPOSE: To describe a case of keratectasia after a deep primary keratotomy during an aborted laser in situ keratomileusis procedure. METHODS: Retrospective, observational case report. RESULTS: In a 47-year-old woman, progressive keratectasia developed after a 90% depth keratotomy during an aborted laser in situ keratomileusis procedure. This case was managed with a rigid gas permeable contact lens and consideration for penetrating keratoplasty. CONCLUSION: Keratectasia is a reported microkeratome-related complication after laser in situ keratomileusis. Appropriate microkeratome assembly and surgeon awareness are necessary to avoid this complication. PMID- 11730661 TI - Chronic cicatrizing conjunctivitis in a patient with ocular cicatricial pemphigoid and fatal Wegener granulomatosis. AB - PURPOSE: To describe a case of chronic cicatrizing conjunctivitis in a patient with ocular cicatricial pemphigoid and Wegener granulomatosis. METHODS: Observational case report. A retrospective study. RESULTS: An 80-year-old man presented with chronic cicatrizing conjunctivitis, peripheral corneal thinning, and Wegener granulomatosis, which were diagnosed by his referring physician based on clinical (recurrent epistaxis, sinus congestion) and histopathologic features of nasal mucosa (granulomatous inflammation, vasculitis). A conjunctival biopsy performed by us disclosed features of active Wegener granulomatosis and ocular cicatricial pemphigoid, which indicate lack of control of both diseases with methotrexate treatment. The patient died of pulmonary complications from Wegener granulomatosis 1 week after our evaluation. CONCLUSION: Ocular cicatricial pemphigoid and Wegener granulomatosis are both potentially fatal autoimmune diseases. Ocular involvement in Wegener granulomatosis indicates poor control of the underlying systemic condition and is a marker for active vasculitis, which indicates the need for treatment with cyclophosphamide. PMID- 11730660 TI - Epstein-Barr virus dacryoadenitis resulting in keratoconjunctivitis sicca in a child. AB - PURPOSE: To describe a case of severe dry eye syndrome in a child. METHODS: Observational case report. The authors describe a 10-year-old male with severe dry eyes who was profoundly disabled by pain and photophobia despite aggressive conventional therapy. Lacrimal gland histology was consistent with the primary Sjogren syndrome, and serologic and immunohistologic evidence supported the hypothesis of Epstein-Barr virus causality. RESULTS: Treatment with systemic acyclovir and cyclosporin A resulted in dramatic and rapid reversal of the profound sicca syndrome and enabled the patient to resume his normal activities. CONCLUSION: Epstein-Barr virus dacryoadenitis should be considered in the differential diagnosis of keratoconjunctivitis sicca in children. Epstein-Barr virus can cause keratoconjunctivitis sicca, which can be treated successfully with acyclovir therapy in addition to suppression of the inflammatory response. PMID- 11730662 TI - Vitreous hemorrhage from a ciliary granuloma associated with Wegener granulomatosis. AB - PURPOSE: To report a case of vitreous hemorrhage from ciliary granuloma in Wegener granulomatosis. METHODS: Interventional case report. An 18-year-old woman with Wegener granulomatosis and episcleral granuloma in her LE had ultrasound biomicroscopy findings of a mass in the pars plana of the ciliary body in the meridian corresponding to the episcleral granuloma. RESULTS: The patient underwent vitrectomy in the LE for subsequent vitreous hemorrhage. Intraoperatively, the mass was diagnosed as a ciliary granuloma at the pars plana. Dense blood clotting around the ciliary granuloma and subretinal exudation at the ora serrata were observed, with no other changes causative for the vitreous hemorrhage. CONCLUSIONS: We report a case of vitreous hemorrhage associated with a ciliary granuloma that was revealed by ultrasound biomicroscopy. Careful observation is necessary in eyes with persistent inflammation in Wegener granulomatosis. PMID- 11730663 TI - Serial axial length measurements in congenital glaucoma. AB - PURPOSE: To illustrate the utility of axial length measurements in the management of congenital glaucoma. METHODS: Interventional case series. Twelve eyes of six consecutive patients with congenital glaucoma were followed with serial axial length measurements. The axial growth patterns were compared with the normal growth curve. RESULTS: Eight (66.7%) of 12 eyes had initial axial lengths longer than the normal range, and 4 eyes (33.3%) were within the normal range. After surgical intervention with good intraocular pressure control in 10 eyes, 3 eyes showed a decrease of axial length and 7 eyes showed no decrease of axial length, but the axial growth pattern paralleled the normal growth curve. After surgical intervention with poor intraocular pressure control, 2 eyes had axial growth pattern greater than the normal pattern. CONCLUSION: With intraocular pressure control in congenital glaucoma, axial length may decrease or the axial growth pattern may parallel the normal growth curve. Axial length measurements may be useful to monitor the control of congenital glaucoma. PMID- 11730664 TI - Choroidal detachment after the use of topical latanoprost. AB - PURPOSE: To report a case of delayed choroidal detachment after treatment with topical latanoprost. METHODS: Interventional case report. Five years after trabeculectomy in a 91-year-old female, a Holmium laser thermal sclerostomy was performed, and a year later, the patient was started on latanoprost.005% every 24 hours and hydrochloride dorzolamide 2% three times daily. RESULTS: Choroidal detachments developed 2.5 years after the sclerostomy; they resolved after discontinuation of the latanoprost and hydrochloride dorzolamide. Subsequently, the patient inadvertently restarted latanoprost. Again, the latanoprost was stopped, with resolution of the choroidal detachments. CONCLUSION: Topical latanoprost may lead to choroidal detachments and hypotony. PMID- 11730665 TI - Migration of intraocular silicone oil into the subconjunctival space and orbit through an Ahmed glaucoma valve. AB - PURPOSE: To report a patient with intraocular silicone oil migration into the subconjunctival space and orbit through an Ahmed glaucoma valve implant. METHODS: Observational case report. A 29-year-old female with diabetes mellitus and proliferative diabetic retinopathy underwent three left eye pars plana vitrectomies and ultimately intraocular silicone oil placement for tractional retinal detachment. Subsequently, she developed neovascular glaucoma and underwent an inferotemporal Ahmed glaucoma tube implant, left eye. RESULTS: Intraocular silicone oil migrated through the Ahmed glaucoma valve into the subconjunctival space and orbit. Left eye evisceration with silicone oil removal was performed for a blind painful left eye and improvement of appearance. CONCLUSION: Silicone oil migration through an Ahmed valve may be observed after retinal detachment repair with intraocular silicone oil. An Ahmed valve may be contraindicated in eyes of patients with intraocular silicone oil. PMID- 11730666 TI - Hypotony maculopathy: improvement of visual acuity after 7 years. AB - PURPOSE: To describe a 79-year-old Caucasian male with a history of hypotony maculopathy for 7 years after cataract extraction with subsequent recovery of normal intraocular pressure and improvement of visual acuity. METHODS: Interventional case report. The patient developed an intraocular pressure ranging from 0 to 5 mm Hg after cataract surgery. He developed hypotony maculopathy, and the visual acuity declined to 20/200. On evaluation 7 years later, a superior cyclodialysis cleft was detected and treated with the argon laser. RESULTS: Closure of the cleft with argon laser was successful. The intraocular pressure stabilized at 17 to 20 mm Hg, and the visual acuity improved to 20/30. CONCLUSION: The development of hypotony maculopathy leads to impairment of visual acuity. This case demonstrates that visual acuity can improve after resolution of the maculopathy even after several years of hypotony. PMID- 11730667 TI - North Carolina macular dystrophy: clinicopathologic correlation. AB - PURPOSE: To describe the clinical and histopathologic findings of a 72-year-old female with North Carolina macular dystrophy. METHODS: Observational case report with histopathologic correlation. Clinical examination includes slit-lamp biomicroscopy, indirect ophthalmoscopy, color fundus photography, and focal electroretinography. Histopathologic examination of the enucleated left eye performed with light microscopy. RESULTS: Light microscopy demonstrated a discrete macular lesion characterized by focal absence of photoreceptor cells and retinal pigment epithelium with attenuation of the Bruch membrane and focal atrophy of the choriocapillaris. Adjacent to the macular lesion, some lipofuscin was identified in the retinal pigment epithelium. CONCLUSION: North Carolina macular dystrophy has both clinical and microscopic appearances of a well demarcated lesion confined to the macula, which involves the retina, pigment epithelium, and choriocapillaris. PMID- 11730668 TI - Isolated late-onset cone-rod dystrophy revealing a familial neurogenic muscle weakness, ataxia, and retinitis pigmentosa syndrome with the T8993G mitochondrial mutation. AB - PURPOSE: To report a late-onset cone-rod dystrophy that revealed a familial neurogenic muscle weakness, ataxia, and retinitis pigmentosa syndrome as a consequence of the T8993G mitochondrial mutation. METHODS: Observational case series. A 42-year-old female disclosed a late-onset retinal dystrophy. The family history revealed that her three sons, one of them deceased at the age of 4, had mental and neurologic impairment of variable severity. The retinal dystrophy of the mother was classified as a cone-rod dystrophy. Retinal dystrophy was subsequently diagnosed in the two surviving sons. Screening for mutation in the mitochondrial DNA (mtDNA) was performed because of the combination of neurologic involvement and retinal dystrophy in this family. RESULTS: Molecular analysis of the mtDNA revealed the ATPase-6 gene T8993G mutation in the mother and the two sons. CONCLUSION: This family illustrates the remarkably variable expression of retinal and systemic manifestations related to the T8993G mutation ranging from an isolated late-onset cone-rod dystrophy to a severe neurodegenerative process with a dramatic outcome. Genetic counseling for retinal dystrophies requires careful evaluation of the familial medical history. PMID- 11730669 TI - Efficacy of (123)N-isopropyl-p-[(123)I]-iodoamphetamine single photon emission computed tomography for the diagnosis of uveal malignant melanoma. AB - PURPOSE: To evaluate the efficacy of single photon emission computed tomography (SPECT) images with N-isopropyl-p-[(123)I]-iodoamphetamine ([(123)I]-IMP) as a radiopharmaceutical for the diagnosis of uveal melanoma. METHODS: Consecutive interventional case series. Single photon emission computed tomography images were obtained after intravenous injection of [(123)I]-IMP in 20 patients with clinically suspected uveal melanoma (16 eyes of 16 patients) or ocular adnexal melanoma (four adnexal tumors of four patients). Histologic findings in seven eyes enucleated for uveal melanoma were analyzed with the SPECT images semiquantitatively evaluated. RESULTS: Eight of 20 eyes with suspected uveal melanoma showed high accumulation of [(123)I]-IMP in the late phase in the area corresponding to the uveal tumor, and the diagnosis of uveal melanoma was confirmed histologically in seven enucleated eyes. Tumor size and histologic features did not reflect the intensity of scintillation counts of [(123)I]-IMP. Twelve patients with no accumulation on the SPECT images were found to have clinically or histologically iris nevus, choroidal nevus, and other intraocular and adnexal conditions. CONCLUSION: N-isopropyl p-[(123)I]-iodoamphetamine SPECT may serve as a sensitive and specific examination for the diagnosis of uveal melanoma. PMID- 11730670 TI - Calcification in choroidal melanoma after transpupillary thermotherapy. AB - PURPOSE: To report the response of calcification in choroidal melanoma after transpupillary thermotherapy. METHODS: Interventional case report. A 66-year-old male with a posterior uveal melanoma measuring 10 x 8 x 4.4 mm underwent infrared diode laser transpupillary thermotherapy three times, each 12 weeks apart. For each treatment, a 3-mm spot size of 1-minute duration was used, and the power setting varied between 450 mW and 500 mW. RESULTS: Two months after the last transpupillary thermotherapy session, A- and B-mode echographic patterns strongly suggestive of intratumor calcification were observed. The tumor regressed to 2.8 mm in thickness, but calcification persisted. Local recurrence or distant metastasis was not detected during a follow-up of 14 months. CONCLUSION: Clinically detectable calcification is extremely rare in choroidal melanomas. Tumor regression with slowly progressive calcification may occur after transpupillary thermotherapy. PMID- 11730671 TI - Choroidal melanoma metastatic to the contralateral choroid. AB - PURPOSE: To report metastasis of choroidal melanoma to the contralateral choroid. METHODS: Interventional case report. In a 49-year-old Caucasian female, a primary choroidal melanoma, left eye, was diagnosed and treated with Ruthenium-106 plaque radiotherapy. The choroidal melanoma showed excellent regression with flattening of the mass and adjacent chorioretinal atrophy. RESULTS: Fifteen years after brachytherapy for choroidal melanoma, left eye, the patient developed a metastatic melanoma to the contralateral choroid as the first sign of metastasis. No history of oculo(dermal) melanocytosis or cutaneous melanoma existed. Systemic evaluation disclosed multiple metastases confined to the liver. CONCLUSION: Metastasis to the contralateral choroid can be the first sign of metastasis from choroidal melanoma. PMID- 11730672 TI - Metastatic adenocarcinoma with rupture through the Bruch membrane simulating a choroidal melanoma. AB - PURPOSE: To report a case of adenocarcinoma metastatic to the choroid with rupture through the Bruch membrane, thus, simulating a choroidal melanoma. METHODS: Interventional case report. Evaluation in a university-based clinic, including a complete eye examination, fluorescein angiography, and ultrasonography; oncological evaluation; and eventual enucleation with histopathological study, including immunohistochemical stains. RESULTS: A 62-year old Hispanic female presented with visual loss, right eye, of short duration because of a choroidal mass with retinal detachment. Ultrasonography showed a dome-shaped lesion with an eccentric collar-button projection and medium internal reflectivity, which suggested a choroidal melanoma. Initial systemic evaluation was negative. Severe pain necessitated enucleation, RE, and histopathology of the choroidal mass demonstrated an adenocarcinoma. Further examination revealed a left, upper lobe, nonsmall cell lung carcinoma. CONCLUSION: Metastatic choroidal tumors may present, although rarely, with collar-button configurations. PMID- 11730673 TI - Interferon therapy for orbital infiltration secondary to Erdheim-Chester disease. AB - PURPOSE: To describe a 55-year-old male with Erdheim-Chester disease with bilateral orbital infiltration and visual loss who was successfully treated with interferon-alpha. METHODS: Interventional case report. RESULTS: The patient was treated with interferon-alpha and had an improvement in his clinical signs, including his visual acuity, after 4 weeks of interferon therapy. CONCLUSION: Interferon-alpha can be effective in the treatment of orbital infiltration secondary to Erdheim-Chester disease. PMID- 11730675 TI - Cataract and phthisis bulbi after laser photoablation for threshold retinopathy of prematurity. PMID- 11730677 TI - Dislocated intraocular lens fixation using intraocular cowhitch knot. PMID- 11730678 TI - New corneal findings in human T-cell lymphotropic virus type 1 infection. PMID- 11730680 TI - Bilateral breast metastases from choroidal melanoma. PMID- 11730696 TI - Immunocytochemical characterization of torsin proteins in mouse brain. AB - Early-onset torsion dystonia is a hyperkinetic movement disorder caused by a deletion of one glutamic acid residue in torsinA, a novel member of the AAA family of ATPases. No mutation has been found so far in the closely related torsinB protein. Little is known about the molecular basis of the disease, and the cellular functions of torsin proteins remain to be investigated. We generated polyclonal anti-peptide antibodies directed against human torsinA and torsinB proteins. In Western blot analysis of mouse brain homogenates, the antibodies specifically recognized 33 kDa endogenous torsinA and 52 kDa endogenous torsinB. Absorption controls showed that labeling was blocked by cognate peptide used for immunization. Immunolocalization studies revealed that torsinA and torsinB were widely expressed throughout the mouse central nervous system. Both proteins were detected in the majority of neurons in nearly all regions. The proteins displayed cytoplasmic distribution, although in some types of neurons localization was perinuclear. Strong labeling of neuronal processes and fibers was detected for both proteins. TorsinA and torsinB have similar CNS distribution, although some differences were observed. Widespread expression suggests that these proteins may play an essential role in normal neuronal functions. The localization of torsinA and torsinB immunoreactivity in neuronal processes points to a potential role for torsin proteins in synaptic functioning. PMID- 11730697 TI - Functional and pharmacological characterization of the modulatory role of serotonin on the firing activity of locus coeruleus norepinephrine neurons. AB - Previous studies, using in vivo extracellular unitary recordings in anaesthetized rats, have shown that the selective 5-HT(1A) receptor antagonist WAY 100,635 suppressed the firing rate of locus coeruleus (LC) norepinephrine (NE) neurons and that this effect was abolished by lesioning 5-HT neurons. In the present experiments, the selective 5-HT(2A) receptor antagonist MDL 100,907, while having no effect on the spontaneous firing activity of LC neurons in controls, was able to restore NE neuronal discharges following the injection of WAY 100,635. The 5 HT(1A) receptor agonist 8-OH-DPAT enhanced the firing activity of NE neurons and this action was entirely dependent on intact 5-HT neurons, unlike the inhibitory effect of the 5-HT(2) receptor agonist DOI. Taken together, these data indicate that 5-HT(2A) but not 5-HT(1A) receptors controlling LC firing activity are postsynaptic to 5-HT neurons. Prolonged, but not subacute, administration of selective 5-HT reuptake inhibitors (SSRIs) produces a decrease in the spontaneous firing activity of LC NE neurons. MDL 100,907 partially reversed this suppressed firing activity of LC neurons in paroxetine-treated rats. Although the alpha(2) adrenoceptor antagonist idazoxan also enhanced the firing activity of NE neurons in paroxetine-treated rats, this increase was similar to that obtained in controls. In conclusion, prolonged SSRI treatment enhances a tonic inhibitory influence by 5-HT on LC neurons through postsynaptic 5-HT(2A) receptors that are not located on NE neurons. A speculative neuronal circuitry accounting for these phenomena on LC NE activity is proposed. PMID- 11730698 TI - Na+ dependent glutamate transporters (EAAT1, EAAT2, and EAAT3) in primary astrocyte cultures: effect of oxidative stress. AB - The Na+ -dependent L-glutamate transporters EAAT1(GLAST), EAAT2 (GLT-1) and EAAT3 (EAAC1) are expressed in primary astrocyte cultures, showing that the EAAT3 transporter is not neuron-specific. The presence of these three transporters was evaluated by RT-PCR, immunoblotting, immunocytochemical techniques, and transport activity. When primary astrocyte cultures were incubated with L-buthionine-(S,R) sulfoximine (BSO), a selective inhibitor of gamma-glutamylcysteine synthetase, the GSH concentration was significantly lower than in control cultures, but the expression and amount of protein of EAAT1, EAAT2 and EAAT3 and transport of L glutamate was unchanged. Oxidative stress was created by adding H(2)O(2) or tert. butyl hydroperoxide (t-bOOH) to the primary astrocyte cultures and cell damage was evaluated by measuring activity of lactate dehydrogenase. Under oxidative stress, GSH levels were significantly lower than in control astrocytes; but the expression and the amount of protein of the three transporters remained unchanged. However, L-glutamate uptake was significantly lower in astrocytes under oxidative conditions when compared to controls. L-Glutamate uptake was not changed in the presence of ascorbate, but was partially recovered in the presence of DTT and GSH ethyl ester. This report emphasizes that oxidative stress and not GSH depletion alters transporter activity without changing transporter expression. PMID- 11730699 TI - Age-related changes in the localization of presenilin-1 in cynomolgus monkey brain. AB - Age-related changes in PS-1 localization were examined in the brains of 22 cynomolgus monkeys ranging in age from embryonic day 87 to 35 years. In embryonic monkey brains, anti-PS-1 antibody N12, which recognizes the PS-1 N-terminal fragment (Ntf) and holo protein, stained immature neuronal cells. In juvenile monkeys, N12 stained large pyramidal neurons, cerebral neocortical neurons, and cerebellar Purkinje's cells. Cytoplasmic staining of these cells was granular in appearance. In aged monkeys, N12 stained neurons in all layers of the neocortex. In contrast, regardless of the age of the animals examined, M5, an anti-PS-1 antibody that specifically recognizes only the PS-1 C-terminal fragment (Ctf), stained neurons in all layers of the neocortex and neurons in the cerebellum. M5 also stained neuropil and white matter, and in aged monkeys, M5 stained swollen neurites of mature senile plaques. Age-related changes in PS-1 expression were further examined using Western blot analysis of mitochondrial, myelin, microsomal, nuclear, synaptosomal, and cytosol fractions isolated from 10 monkey brains ranging in age from embryonic day 87 to 32 years. In all brains, Ntf and Ctf were expressed most abundantly in the microsome fraction. The amount of PS-1 in the nuclear fraction dramatically increased with age. We conclude that the transport of PS-1 diminished with age and that PS-1 fragments accumulated in endoplasmic reticulum (ER) associated with the nuclear membrane. PMID- 11730700 TI - Neuroprotective effects of the N-terminal tripeptide of IGF-1, glycine-proline glutamate, in the immature rat brain after hypoxic-ischemic injury. AB - Insulin growth factor 1 (IGF-1) has an important role in brain development and is strongly expressed during recovery after a hypoxic-ischemic injury. Some of its central actions could be mediated through the N-terminal tripeptide fragment of IGF-1: Gly-Pro-Glu (GPE). The neuroprotective properties of GPE given after a moderate injury in the developing rat brain were evaluated and the binding sites of [(3)H]GPE characterised by autoradiography. After right unilateral injury, GPE or vehicle (V) was injected in the right lateral ventricle (i.c.v.) or in the peritoneal cavity (i.p.) of 21-day-old rats. The percentage of surviving neurons in CA1-2 of the hippocampus was higher in the animals treated with 30 microg of GPE i.c.v. (V: 7.7+/-4.9%, GPE: 26.4+/-7.5%, P=0.02) and 300 microg i.p. (V: 30.2+/-9.1%, GPE: 68.8+/-10.6%, P=0.02) than in animals receiving vehicle. I.p. injection of 300 microg of GPE (V: 78.4+/-7.5%, GPE: 88.4+/-3.2%, P=0.04) was also neuroprotective in the lateral cortex. I.c.v. injection of [(3)H]GPE suggested binding to glial cells in the white matter tracts, the cortex and striatum as opposed to neurons. Although the precise mode of action of GPE is unknown, this study suggests that local administration of GPE is neuroprotective after brain HI injury via glial cells. In addition, systemic administration of GPE showed a more widespread neuroprotective effect. GPE may represent a complementary pathway for central and systemic IGF-1's antiapoptotic effects. PMID- 11730701 TI - 6-Hydroxydopamine-lesioning of the nigrostriatal pathway in rats alters basal ganglia mRNA for copper, zinc- and manganese-superoxide dismutase, but not glutathione peroxidase. AB - The effects of nigrostriatal pathway destruction on the mRNA levels of copper, zinc-dependent superoxide dismutase (Cu,Zn-SOD), manganese-dependent superoxide dismutase (Mn-SOD), and glutathione peroxidase in basal ganglia of adult rat were investigated using in situ hybridization histochemistry and oligodeoxynucleotide (single-stranded complementary DNA) probes. The 6-hydroxydopamine (6-OHDA) induced destruction of the nigrostriatal pathway resulted in contralateral rotation to apomorphine and a marked loss of specific [(3)H]mazindol binding in the striatum (93%; P<0.05) and of tyrosine hydroxylase mRNA in substantia nigra pars compacta (SC) (93%; P<0.05) compared with control rats. Levels of Cu,Zn-SOD mRNA were decreased in the striatum, globus pallidus, and SC on the lesioned side of 6-OHDA-lesioned rats compared with sham-lesioned rats (P<0.05). Levels of Mn SOD mRNA were increased in the nucleus accumbens (P<0.05), but decreased in the SC (P<0.05) on the lesioned side of 6-OHDA-treated rats compared with sham lesioned rats. Lesioning with 6-OHDA had no effect on glutathione peroxidase mRNA levels in any region of basal ganglia examined. The significant changes in Cu,Zn SOD and Mn-SOD mRNA indicate that SOD is primarily expressed by dopaminergic neurons of the nigrostriatal pathway, and that the Mn-SOD gene appears to be inducible in rat basal ganglia in response to both physical and chemical damage 5 weeks after 6-OHDA-lesioning. These findings may clarify the status of antioxidant enzymes, particularly Mn-SOD, in patients with Parkinson's disease and their relevance to disease pathogenesis. PMID- 11730702 TI - Intracellular calcium and outer hair cell electromotility. AB - The influence of increased intracellular calcium level on outer hair cell (OHC) electromotility was examined by means of transcellular electrical stimulation in a partitioning microchamber. Electromotile activity was measured before and after application of the calcium ionophore ionomycin, which promotes the inflow of extracellular calcium, as well as its release from intracellular calcium stores. The ionomycin solvent, dimethyl sulphoxide (DMSO), by itself elicited a significant decrease in the magnitude of OHC electromotility. The DMSO effect was counteracted by 10 microM ionomycin and was reversed by 50 microM ionomycin. The increase in electromotility is partially mediated by a calmodulin-dependent mechanism, since W7, a calmodulin antagonist, attenuated the 50 microM ionomycin induced motility increase. Our results suggest that the electromotility magnitude increase in isolated OHCs due to ionomycin is a calcium/calmodulin-dependent phenomenon. PMID- 11730703 TI - Dynamic changes in luteinizing hormone releasing hormone transcriptional activity are associated with the steroid-induced LH surge. AB - Luteinizing hormone releasing hormone (LHRH) gene transcription was examined in ovariectomized female rats on the day of a steroid-induced LH surge using the RNase protection assay. LHRH mRNA levels were measured in cytosolic extracts, and LHRH primary transcript levels were measured in nuclear extracts prepared from tissue fragments that contained the organum vasculosum of the lamina terminalis (OVLT) and the preoptic area (POA). Measurements of both mature and primary transcript levels demonstrated modest but significant changes over time. Alterations in LHRH primary transcript levels preceded changes in levels of mature mRNA suggesting a delay in the detectable response of the cytoplasmic pool of LHRH mRNA to changes in gene transcription at this time. When viewed in relation to circulating LH titers, LHRH primary transcript levels were high prior to the start of the LH surge and after peak levels of LH were attained, and they declined during the ascending phase of the LH surge. These findings suggest a potential role for increased LHRH gene transcription in the accumulation of LHRH prior to the start of the LH surge and in the replenishment of LHRH stores depleted during the surge. Moreover, the decrease in LHRH gene transcription during the ascending phase of the LH surge may be important for limiting surge duration. The data presented are consistent with a role for dynamic changes in LHRH transcriptional activity in modulating parameters of the steroid-induced LH surge and in replenishing the releasable pool of this essential decapeptide. PMID- 11730704 TI - Intra-prefrontal cortex injections of SCH 23390 influence nucleus accumbens dopamine levels 24 h post-infusion. AB - The dopaminergic pathway from the ventral tegmental area (VTA) to the nucleus accumbens (NAcc) is well known to be involved in the reinforcing properties of many drugs of abuse. The medial prefrontal cortex (mPFC) has been shown to exhibit significant influence over activity in this pathway, and has also been implicated in drug abuse. The present experiment investigated the ability of D1 activity in the mPFC to influence accumbal dopamine levels. NAcc dopamine (DA) was monitored before, immediately after, and 24 h following mPFC infusion of a D1 agonist (SKF 38393), D1 antagonist (SCH 23390), or a vehicle solution. Immediately following infusion of dopaminergic agents or vehicle, no significant changes in accumbal DA were observed. However, 24 h following infusion of the antagonist but not the agonist, significant elevations of accumbal DA were observed. Since elevated NAcc DA was only observed 24 h after treatment, these results provide evidence that long-term neural adaptations can be induced by transient neuropharmacological treatment. PMID- 11730705 TI - Vomeronasal organ: electrical stimulation activates Fos in mating pathways and in GnRH neurons. AB - Electrical stimulation of the vomeronasal organ in male hamsters activated Fos expression in neurons of the chemosensory pathways, as in experiments where animals were stimulated with female chemical stimuli. Fos was also activated in gonadotropin hormone releasing hormone (GnRH, or LHRH) neurons in the rostral medial preoptic region of the brain, a possible substrate for GnRH influence on chemosensory-dependent reproductive behavior. PMID- 11730706 TI - Differential gating of slow postsynaptic and high-frequency spike-like components in human somatosensory evoked potentials under isometric motor interference. AB - Human cortical somatosensory evoked potentials (SEP) can be modified by concomitant motor tasks ('gating'), through peripheral occlusion and/or central mechanisms. The present study aimed (1) at refining earlier results concerning motor-gating of the primary cortical EPSP-related N20 response after electric median nerve stimulation, and (2) at providing first data on motor-gating of the 600 Hz SEP wavelet burst which occurs superimposed onto N20 and primarily reflects repetitive cerebral population spikes. In 12 healthy subjects median nerve SEP were elicited, using electrical stimuli with intensities below, at and above motor threshold, under either rest or an isometric fist clenching task. Amplitude and latency modifications were analysed for the peripheral compound action potential (CAP), low-frequency SEP components (N20, P25, N35 and P70) and the high-frequency burst. While the peripheral CAP remained unchanged, isometric motor innervation significantly attenuated N20, P25 and P70 amplitudes and shortened peak latencies progressively for all components after N20. In contrast, the high-frequency 600 Hz burst was modulated neither in amplitude nor in latency. Regular amplitude recruitment occurred for all components independent from the motor task, excluding channel saturation as an explanation for gating. We suggest that SEP gating under isometric motor innervation is a central process which selectively operates on specific SEP components and could partly reflect an "efference copy" mechanism. PMID- 11730707 TI - Effects of cholecystokinin tetrapeptide (CCK(4)) and anxiolytic drugs on the electrically evoked [(3)H]5-hydroxytryptamine outflow from rat cortical slices. AB - The outflow of [(3)H]5-hydroxytryptamine ([(3)H]5-HT) from electrically stimulated rat cortical slices was measured to ascertain the modulatory role of endogenous cholecystokinin (CCK) on the amine outflow and to test the hypothesis that different anxiolytic compounds inhibit 5-HT secretion. The [(3)H]5-HT outflow evoked at 10 Hz was increased up to +30% by CCK(4) 300-1000 nM, the effect being prevented by the CCK(B) receptor antagonist GV 150013, 3 nM. The limited sensitivity to CCK(4) seemed to depend on 5-HT auto-receptor feedback because pre-treatment with 100 nM methiothepin enhanced the [(3)H]5-HT outflow and lowered the CCK(4) threshold concentration from 300 to 30 nM. In addition, pre-treatment with 1 microM bacitracin to inhibit CCK metabolism increased [(3)H]5-HT efflux. This effect was concentration-dependently counteracted by GV150013 suggesting the presence of an endogenous CCK positive modulation. GV150013 30 nM, the 5-HT(1A) partial agonist buspirone 300 nM and the GABA(A) receptor modulator diazepam 10 nM, known to have anxiolytic properties, all significantly reduced the [(3)H] amine outflow from cortical slices by about 20%. This inhibition depended on their interaction with their respective receptors, which seemed to restrain the activity of functionally interconnected glutamatergic interneurones. In fact, APV (50 microM) and NBQX (10 microM) prevented the effect of the anxiolytic compounds. Thus, anxiolytic drugs with different receptor targets can reduce 5-HT outflow by dampening the glutamatergic signal, and in turn, the secretory process of the serotonergic nerve ending. PMID- 11730708 TI - Electrophysiological effects of sustained delivery of CRF and its receptor agonists in hippocampal slices. AB - The corticotropin-releasing factor (CRF) is a hypothalamic peptide that regulates the release of adrenocorticotropic hormone (ATCH) and of beta-endorphin. It has been suggested that it modulates learning and memory processes in rat. However, the electrophysiological effects that CRF produces on hippocampal neurons have been so far little investigated. In particular, the effects of CRF on long-term potentiation (LTP), a phenomenon which is thought to be the substrate of memory processes, are unknown. We studied the effects of sustained administration of CRF and of two of its receptor agonists on basal neuronal activity and on in vitro hippocampal LTP. The two receptor agonists were D-Glu-20-CRF and D-Pro-5-CRF, selective for the CRF-R1 and the CRF-R2 receptors, respectively. We found that CRF, D-Pro-5-CRF and D-Glu-20-CRF at the concentration of 1 nM diminish the amplitude of hippocampal population spike and prevent the onset of LTP. Higher concentrations of CFR have less depressing effects on neuronal activity, yet they still prevent the occurrence of LTP. PMID- 11730710 TI - Social attraction between rats in open field: long-term consequences of kindled seizures. AB - Kindling of the amygdaloid complex in rats results in an enhanced emotionality frequently expressed by an elevated anxiety and defensive attitude toward other animals. Defensive attitude may have important consequences in social context and if tested in a large space it may eventually lead to social withdrawal. To test this hypothesis, rats were subjected to daily kindling sessions and their behavior was compared to implanted-sham and intact rats. Blood was collected after selected kindling trials for assessment of corticosterone response. Behavioral tests started 1 month after the last kindling trial and consisted of two open field sessions. A solitary rat was tested in the 1st session and pair of rats was tested simultaneously in the second session. Results showed that kindling changed the balance between exploration and occupation of a home base (HB) in the open field, in favor of higher preference of the home base occupancy. These results were apparent only during the social session leading to the conclusion that rats preferred to stay in the home base to maximize the proximity to a partner rat. This was supported by the observation that by increasing the occupancy of the HB, the kindled rats accomplished the longest concurrent presence with the partner rat in the common HB. We discuss the level of inter rats aggression as a factor defining whether the anxious kindled rats will respond with increased or decreased social attraction in the open field test. PMID- 11730709 TI - Nociceptin/orphanin FQ into the rat periaqueductal gray decreases the withdrawal latency to heat and loading, an effect reversed by (Nphe(1))nociceptin(1 13)NH(2). AB - The present study investigated the effect of intraperiaqueductal grey injection of nociceptin/orphanin FQ (N/OFQ) and an antagonist (Nphe(1))nociceptin(1 13)NH(2) on the hindpaw withdrawal response to thermal and mechanical stimulation in rats. N/OFQ (5 nmol) significantly decreased the nociceptive thresholds in both tests and 1, 5 and 10 nmol of (Nphe(1))nociceptin(1-13)NH(2) significantly reversed this effect in a dose dependent way. Our results demonstrate, that N/OFQ has a nociceptive action, possibly through inhibition of PAG neurons. This effect is blocked by the antagonist (Nphe(1))nociceptin(1-13)NH(2) probably via ORL1 receptors in the periaqueductal grey. PMID- 11730711 TI - Temporal and spatial differences of PSA-NCAM expression between young-adult and aged rats in normal and ischemic brains. AB - Highly polysialylated neural cell adhesion molecule (PSA-NCAM) is transiently expressed specifically in newly generated cells, and is important for migration and neurite outgrowth. To investigate the effect of aging on the migration of neural stem cell (NSC) after brain ischemia, the spatiotemporal expressions of immunoreactive PSA-NCAM were examined at 4 h or 1, 3 or 7 days after 90 min of middle cerebral artery occlusion (MCAO) in the young-adult or aged rats. In the sham control brain, PSA-NCAM staining was slightly observed both in dorsal and ventral parts of subventricular zone (SVZ) in the aged brain, but only in the dorsal part of SVZ in the young brain. After transient MCAO, immunoreactivity for PSA-NCAM increased in the number and the intensity in SVZ ipsilateral to MCAO in the young-adult brains and became the peak at 1 day, while that was at 3 days in the aged brains. These findings suggest that PSA-NCAM was located in different spatial distribution in normal condition between young and old rats. PSA-NCAM was induced after ischemia, and the temporal expression was also different after transient MCAO between young and older rats. PMID- 11730712 TI - Protective effects of treadmill training on infarction in rats. AB - This study was undertaken to determine the protective effects of treadmill training on brain ischemic lesions caused by middle cerebral artery (MCA) occlusion in male rats. Rats were divided into four groups: control, 1-week treadmill pre-training, 2-week treadmill pre-training, and 4-week treadmill pre training. Cerebral infarction was induced by MCA occlusion for 60 min, followed by reperfusion. After 24 h, rats were killed and brain slices were then stained to assess lesion size. Treadmill training at least for 2 weeks can reduce the infarction size and edema caused by MCA occlusion (P<0.01). The present study provides evidence that treadmill training reduces ischemic brain damage in an animal model of cerebral ischemia. PMID- 11730713 TI - Leukemia inhibitory factor by systemic administration rescues spinal motor neurons in the SOD1 G93A murine model of familial amyotrophic lateral sclerosis. AB - Leukemia inhibitory factor (LIF) is a survival factor for motoneurons. In this study we investigated whether intense systemic LIF therapy prevents the loss of lumbar motoneurons in the transgenic SOD1 G93A mouse model of familial amyotrophic lateral sclerosis. Treatment involved daily 25 microg/kg intraperitoneal injection for a period of 6 weeks starting at 70 days of age. Using the unbiased optical dissector technique, significant rescue of motoneurons in the LIF-treated group (3809+/-455) was found compared to the vehicle group (1085+/-140). PMID- 11730714 TI - Increased cAMP immunostaining in cerebral vessels in Alzheimer's disease. AB - Increasing evidence suggests that up-regulation of the cAMP-second messenger system is implicated in Alzheimer's disease neurodegeneration. Since previous studies reported an increased level of cAMP in microvessels of Alzheimer's patients compared with those from non-demented elderly controls, we have carried out an immunohistochemical study to compare cAMP immunostaining in brain vessels from patients with dementia and neuropathological criteria of Alzheimer's disease (n=5) with those of age-matched patients (n=10). We have also included a control group of adult patients (n=5) to evaluate the role of aging separate from the effects of dementia. Our results demonstrated an increased cAMP immunostaining in cerebral cortical and meningeal vessels from Alzheimer's patients compared to nondemented elderly and adult controls. Vascular cAMP immunostaining was mainly observed in frontal and temporal cortex, the hippocampus being the region that showed the more intense and widespread vascular cAMP immunostaining. We also observed a conspicuous vascular beta-amyloid immunostaining specifically in those vessels that showed the highest cAMP immunostaining. We suggest that increased vascular cAMP immunostaining is mainly localised in the selectively vulnerable targets of neurodegeneration that characterise AD. Moreover, the co immunolocalisation of cAMP and beta-amyloid protein in cerebral vessels of patients with AD suggests a possible role of cAMP up-regulation in the accumulation of those amyloidogenic peptides. PMID- 11730715 TI - Early effects on restoration of evoked field potentials in the hippocampal CA(1) region after reversible hypoxia/hypoglycemia by the radical scavenger N-tert. butyl-alpha-phenylnitron. AB - In transverse hippocampus slices a short period of hypoxia/hypoglycemia induced by perfusion with an O(2)/glucose-free medium caused early loss and incomplete restoration of evoked field potentials in the CA(1) region. In the present study a search was made for whether the formation of free oxoradicals immediately after starting the hypoxic phase could be part of the breakdown and incomplete restoration of the excitatory potentials (EPs). It was shown that preincubation and postischemic incubation with the radical scavenger PBN did not prevent the potential breakdown but significantly enhances potential restoration, even when PBN was added to the perfusion medium 40 min after hypoxia. Thus, free oxoradicals may damage membrane constituents such as receptors or channel proteins at a very early phase, before neuronal death is pronounced. The results also show that treatment with radical scavengers has a beneficial effect on early hypoxic damage. PMID- 11730716 TI - The protective action of nefiracetam against electrophysiological and metabolic damage in the hippocampus after deprivation of glucose and oxygen. AB - The present study examined the effect of nefiracetam on ischemic brain damage by monitoring population spikes (PSs) in the dentate gyrus of guinea pig hippocampal slices; assaying high-energy phosphates (ATP and CrP) in guinea pig hippocampal slices; and monitoring whole-cell membrane-currents and intracellular Ca(2+) levels in cultured hippocampal neurons. Twenty-minute ischemic insult to slices, i.e., deprivation of glucose and oxygen from artificial cerebrospinal fluid, abolished PSs. As compared with only 35% recovery of the PS amplitude for control, PS amplitude reversed to 65% of basal levels 40 min after returning normal conditions by treatment with nefiracetam (0.01 microM). Ischemic insult reduced the levels of adenosine triphosphate (ATP) and creatine phosphate (CrP) in slices, and when returned to normal conditions, recovering to 70 and 85% of basal values, respectively, 30 min after returning normal conditions. Nefiracetam (0.01 microM) facilitated the recovery of ATP and CrP, reaching 110 and 140% of basal values, respectively. Nefiracetam inhibited N-methyl-D-aspartate (NMDA) evoked currents to 35% of basal amplitudes. Likewise, nefiracetam (0.01 microM) inhibited intracellular Ca(2+) rise through NMDA receptor channels to 30% of basal levels. The results of the present study, thus, suggest that nefiracetam has the potential to protect against ischemic brain damage, possibly in part by preventing from accumulation of intracellular calcium through NMDA receptor channels. PMID- 11730718 TI - Modulation of miniature inhibitory postsynaptic currents by isoflurane in rat dissociated neurons with glycinergic synaptic boutons. AB - The effects of a volatile anesthetic, isoflurane, on glycinergic miniature inhibitory postsynaptic currents (IPSCs) were investigated in mechanically dissociated rat trigeminal nucleus neurons with intact glycinergic interneuronal presynaptic nerve terminals. The nystatin-perforated patch recording configuration was used to record the miniature IPSCs under voltage-clamp conditions. Isoflurane shifted in a parallel fashion the glycine (Gly) concentration-response curve of enzymatically dissociated neurons to the left without changing the maximum response. Isoflurane reversibly increased the frequency of the miniature IPSCs and prolonged the decay time constant without affecting the mean amplitude. The increase in the frequency of miniature IPSCs in the presence of isoflurane was also observed in Ca(2+)-free external solution. Thapsigargin prohibited the facilitatory effect of isoflurane on the miniature IPSC frequency. It is concluded that isoflurane increases the Ca(2+) concentration in the glycinergic presynaptic nerve terminal by enhancing the release and/or suppressing the uptake of Ca(2+) into stores. PMID- 11730720 TI - Effects and mechanisms of aloe-emodin on cell death in human lung squamous cell carcinoma. AB - Aloe-emodin (1,8-dihydroxy-3-(hydroxymethyl)-anthraquinone) is an active component from the root and rhizome of Rheum palmatum. The study investigated the effects and mechanisms of aloe-emodin-induced cell death in human lung squamous cell carcinoma cell line CH27. Aloe-emodin (40 microM)-induced CH27 cell apoptosis was confirmed by DNA fragmentation (DNA ladders and sub-G(1) formation). Aloe-emodin-induced apoptosis of CH27 cells involved modulation of the expression of Bcl-2 family proteins, such as BclX(L), Bag-1, and Bak, and was associated with the translocation of Bak and Bax from cytosolic to particulate fractions. Aloe-emodin-treated CH27 cells had an increased relative abundance of cytochrome c in the cytosolic fraction. Results demonstrated that the activation of caspase-3, caspase-8, and caspase-9 is an important determinant of apoptotic death induced by aloe-emodin. These results suggest that aloe-emodin induces CH27 cell death by the Bax and Fas death pathway. PMID- 11730719 TI - Cannabinoid CB(1) receptor expression, activation and detection of endogenous ligand in trabecular meshwork and ciliary process tissues. AB - Elevated intraocular pressure is the primary risk factor for glaucoma. Cannabinoids interact with molecular targets in the eye and lower intraocular pressure by an unknown mechanism. The purpose of the present study was to examine eye tissues for functional cannabinoid receptors of the neuronal, CB(1) class, and an endogenous ligand, anandamide. The trabecular meshwork and ciliary processes are the primary structures of the eye that contribute to intraocular pressure and thus were our focus. Total RNA, frozen sections, cellular membranes and primary cultures of cells were prepared from both bovine and cadaveric human tissues. Using cannabinoid CB(1) receptor-specific oligodeoxynucleotide primers, cannabinoid CB(1) receptor antiserum, and cannabinoid-specific compounds (CP 55,940, WIN55,212-2 and SR-141716A), the presence of cannabinoid CB(1) receptors in ciliary processes and trabecular meshwork was determined. Using reverse transcription-polymerase chain reaction, we identified mRNA encoding cannabinoid CB(1) receptor protein in ciliary process and trabecular meshwork cells. Specific binding of anti-CB(1) immunoglobulin-G in tissue sections localized cannabinoid CB(1) receptor protein to the non-pigmented epithelial cells of the ciliary process and cells of the trabecular meshwork. While CP-55,940 and WIN55,212-2 failed to stimulate [(35)S]GTP gamma S binding in membrane preparations from trabecular meshwork and ciliary process, CP-55,940 significantly stimulated whole cell [(35)S]GTP gamma S binding by 51% over basal in ciliary process epithelial cells and 69% over basal in trabecular meshwork cells permeabilized with 5 microM digitonin (p<0.001). Specificity of agonist stimulation was verified by complete blockade with the specific cannabinoid CB(1) receptor antagonist, SR-141716A. Moreover, activation of cannabinoid CB(1) receptors by CP-55,940 resulted in a 2.3+/-0.3 and 1.7+/-0.3-fold stimulation of cAMP accumulation in trabecular meshwork and ciliary process cells, respectively (p<0.01). Lastly, anandamide was detected in human trabecular meshwork (3.08 pmol/g), ciliary process (49.42 pmol/g) and neurosensory retinal (4.48 pmol/g) tissues. These data, for the first time, demonstrate in a single study the presence of both CB(1) mRNA and protein in trabecular meshwork and ciliary processes from two different species. Activation of heterotrimeric G-proteins and stimulation of cAMP accumulation by cannabinoids in vitro suggest that their intraocular pressure-lowering effects in vivo result from activation of cannabinoid CB(1) receptors in the trabecular meshwork and increase aqueous outflow. PMID- 11730721 TI - Transport characteristics of grepafloxacin and levofloxacin in the human intestinal cell line Caco-2. AB - Transport characteristics of grepafloxacin and levofloxacin across the apical membrane of Caco-2 cells were examined. Both grepafloxacin and levofloxacin uptakes increased rapidly, and were temperature-dependent. Grepafloxacin and levofloxacin uptakes showed concentration-dependent saturation with Michaelis constants of 3.9 and 9.3 mM, respectively. Uptake of grepafloxacin and levofloxacin increased in Cl(-)-free and ATP depleted conditions, suggesting the involvement of an efflux transport system different from the uptake mechanism. However, cyclosporin A, a typical inhibitor of P-glycoprotein, did not affect the uptake of these drugs. Unlabeled grepafloxacin, unlabeled levofloxacin and quinidine inhibited the uptake of grepafloxacin and levofloxacin under Cl(-)-free conditions. Tetraethylammonium, cimetidine, p-aminohippurate, probenecid, amino acids, beta-lactam antibiotic or monocarboxylates did not inhibit the uptake of grepafloxacin and levofloxacin under the same conditions. In conclusion, our results suggested that grepafloxacin and levofloxacin uptakes were mediated by a specific transport system distinct from those for organic cations and anions, amino acids, dipeptides and monocarboxylates. PMID- 11730722 TI - Regional mapping of low-affinity kainate receptors in mouse brain using [(3)H](2S,4R)-4-methylglutamate autoradiography. AB - Recent data indicate that (2S,4R)-4-methylglutamate is a selective agonist for low affinity (GluR5 and GluR6) kainate receptor subunits. In the present study, we have employed [(3)H](2S,4R)-4-methylglutamate to examine low affinity kainate receptor distribution in mouse brain. [(3)H](2S,4R)-4-Methylglutamate labelled a single site in murine cerebrocortical membranes (K(d)=9.9+/-2.7 nM, B(max)=296.3+/-27.1 fmol mg protein(-1)). The binding of 8 nM [(3)H](2S,4R)-4 methylglutamate was displaced by several non-NMDA receptor ligands (K(i)+/ S.E.M.): domoate (1.1+/-0.2 nM)>kainate (7.1+/-1.1 nM) >> L-glutamate (187.6+/ 31.9 nM) >> (S)-alpha-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) (>50 microM). [(3)H](2S,4R)-4-Methylglutamate autoradiography revealed a widespread regional distribution of low affinity kainate receptors. Highest binding densities occurred within deep layers of the cerebral cortex, olfactory bulb, basolateral amygdala and hippocampal CA3 subregion. Moderate labelling was also evident in the nucleus accumbens, dentate gyrus, caudate putamen, hypothalamus and cerebellar granule cell layer. These data show that [(3)H](2S,4R)-4-methylglutamate is a useful radioligand for selectively labelling low affinity kainate receptors. PMID- 11730723 TI - Agonists of proteinase-activated receptor 2 excite guinea pig ileal myenteric neurons. AB - The effects of proteinase-activated receptor 2 (PAR2) agonists on the electrical properties of intact guinea pig ileal myenteric neurons were measured with intracellular microelectrodes. Approximately 52% of AH neurons and 41% of S neurons responded to pressure ejection of SLIGRL-NH(2) or trypsin with a prolonged depolarization that was often accompanied by increased excitability. When added to the bathing solution, trypsin caused a concentration-dependent depolarization of responding neurons with an estimated EC(50) value of 87 nM. Collectively, these novel observations indicate that PAR2 excites a proportion of myenteric neurons, which may contribute to dysmotility during intestinal inflammation. PMID- 11730724 TI - Effects of ketamine on different types of anxiety/fear and related memory in rats with lesions of the median raphe nucleus. AB - The aim of the present study was to determine the involvement of the median raphe serotonergic system in the effects of ketamine on anxiety behaviours and related memory. The effects of ketamine pretreatment (3 and 10 mg/kg, i.p.) on three types of fear-motivated behaviours, unconditioned one-way escape, conditioned avoidance and freezing were tested. Experiments were performed with the inhibitory avoidance apparatus in rats with ibotenic acid lesions of the median raphe nucleus. It was found that 10 mg/kg ketamine had an anxiogenic-like effect on one-way escape type of fear and anxiolytic-like effect on conditioned freezing related fear; these effects were unaffected by median raphe lesions. Both ketamine doses impaired freezing-related fear memory. Ketamine (10 mg/kg) also produced an anxiolytic-like effect on avoidance type of fear and impaired avoidance memory. The median raphe lesions attenuated the anxiolytic action of the drug on the avoidance type of fear and prevented ketamine-induced avoidance memory impairment. These results suggest that the anxiolytic-like effect of ketamine on avoidance-type fear is mediated through the median raphe serotonergic system. PMID- 11730725 TI - Renal vascular reactivity to vasopressin in rats with diabetes mellitus. AB - We evaluated how renal vascular reactivity to vasopressin changes when nitric oxide (NO) synthesis varies, as has been reported to occur in the course of insulin-dependent diabetes mellitus. Renal vasoconstrictor responses to vasopressin were obtained in young and older Sprague-Dawley control rats (3 and 10 months old) and in age-matched diabetic rats that had been treated with streptozotocin (60 mg/kg i.v.) at the age of 2 months. In young rats, vasopressin (3-1000 ng/kg/min i.v.) induced in vivo a dose-dependent decrease in renal blood flow, which was diminished in streptozotocin diabetic rats (P<0.05). Similarly, in in vitro perfused kidneys, the concentration-response curve for vasopressin (0.03-10 nM) was shifted 3-fold to the right in kidneys isolated from young diabetic rats (P<0.05). This shift was abolished in the presence of an inhibitor of nitric oxide synthesis, N(G)-nitro-L-arginine (100 microM), in the perfusate. In 10-month-old rats, the in vivo renal vasoconstrictor dose-response curve to vasopressin was shifted 10-fold to the left as compared to that for young rats (P<0.001). This shift was similar in both control and diabetic rats. In conclusion, the present study documented the existence of hyporesponsiveness to vasopressin in the early stage of diabetes, possibly related to nitric oxide overproduction. In contrast, renal vascular hyperreactivity to vasopressin occurs with aging, whether the rats are diabetic or not. PMID- 11730726 TI - Absence of exacerbation of myocardial stunning in anesthetized dogs treated with KAD-1229, a novel hypoglycemic agent. AB - The effect of (+)-momocalcium bis[(2S,3a,7a-cis)-alpha-benzylhexahydro-gamma-oxo 2-isoindolinebutyrate]dihydrate (KAD-1229), a novel hypoglycemic agent with a chemical structure different from that of the sulfonylureas, on myocardial stunning was assessed in anesthetized dogs by comparison with that of glibenclamide, a sulfonylurea. Even though their hypoglycemic effects were of similar magnitude, glibenclamide (1 mg/kg, i.v.), but not KAD-1229, exacerbated the myocardial stunning induced by occlusion/reperfusion of the descending coronary artery. In a receptor-binding experiment, unlabeled glibenclamide completely inhibited [(3)H]glibenclamide binding to the myocardium, but KAD-1229 did not. These results suggest that the difference in binding properties of KAD 1229 and glibenclamide toward cardiac sulfonylurea receptors is one of the causes of their different effects on myocardial stunning. It is likely that KAD-1229 is highly specific for pancreatic sulfonylurea receptors and is speculated to be a safer hypoglycemic agent than, at least, glibenclamide. PMID- 11730727 TI - Effects of GABA(B) receptor antagonists on spontaneous and on GABA-induced mechanical activity of guinea-pig smooth muscle preparations. AB - The majority of GABA(B) receptor antagonists have been based on alterations of the acidic moiety of gamma-aminobutyric acid (GABA) or baclofen, such as the first selective antagonist phaclofen. More recently, a new structural class of compounds derived by p-alkyl substitution in the phosphinic analog of GABA, such as CGP35348 (3-amino-propyl-(diethoxymethyl)-phosphinic acid), have been introduced as GABA(B) receptor antagonists. The present study examine the influence of a series of structurally related phosphinic acid analogues on mechanical activity and their effect on GABA-induced reactions in ileal smooth muscle. In our experiments, GABA exerted a biphasic contractile-relaxation effect with pronounced dose-dependent characteristics. 3-[[1-(S)-(3,4-Dihydrophenyl) ethyl]amino]-2-(S)-hydroxy-propyl]-(phenylmethyl)-phosphinic acid hydrochloride (CGP55845A) induced prolonged relaxation without changing the phasic activity of the ileum preparations. [3-[1-R-[[2-(S)-hydroxy-3-[hydroxy-4-methoxyphenyl] methyl]-phosphinyl]-propyl]-aminoethyl]-benzoic acid (CGP62349) did not change the mechanical activity of smooth muscle preparation. Trans 3-[6-[[Cyclo hexylmethyl-hydroxy-phosphinyl]-methyl]-3-morpholinyl]-benzoic acid (CGP71982) itself induced smooth muscle contractions. GABA(B) receptor antagonists decreased concentration-dependently the relaxation phase of the action of GABA from 50% to 90%. Their effect on the contractile phase of the action of GABA was quite different-CGP55845A decreased it dose-dependently, whereas CGP62349 and CGP71982 did not change it significantly. These findings prompted us to assume that the GABA(B) receptor antagonists studied, being phosphinic analogues, probably act on GABA(B) receptors in guinea-pig ileum smooth muscles. PMID- 11730728 TI - Squalene synthase inhibitors reduce plasma triglyceride through a low-density lipoprotein receptor-independent mechanism. AB - Inhibitors of squalene synthase are considered to be candidate drugs to reduce both plasma cholesterol and triglyceride. However, little is known about the mechanism of squalene synthase inhibitor-specific effect on plasma triglyceride. In this study, we confirmed the triglyceride-lowering effect of ER-27856, a potent squalene synthase inhibitor prodrug, in rhesus monkeys. To determine the role of low-density lipoprotein (LDL) receptor in the triglyceride-lowering effect of squalene synthase inhibitors, we intravenously administered ER-28448, the active form of ER-27856, to Watanabe heritable hyperlipidemic (WHHL) rabbits for 4 days. In heterozygotes, ER-28448 reduced plasma cholesterol and triglyceride by 52% and 37%, respectively. In homozygous rabbits, in contrast, ER 28448 lowered plasma triglyceride by 40% but did not lower plasma cholesterol. Orally administered ER-27856 reduced plasma triglyceride in homozygous animals but atorvastatin and bezafibrate did not. In hepatocytes isolated from homozygous WHHL rabbits, squalene synthase inhibitors but not atorvastatin reduced triglyceride biosynthesis. These data demonstrate that squalene synthase inhibitors reduced plasma triglyceride through an LDL receptor-independent mechanism, which was distinct from that of the triglyceride-lowering action of atorvastatin or bezafibrate. The reduction of hepatic triglyceride biosynthesis may play an important role in the hypotrigyceridemic action of squalene synthase inhibitors. PMID- 11730729 TI - Effect of fenoterol-induced constitutive beta(2)-adrenoceptor activity on contractile receptor function in airway smooth muscle. AB - In the present study, we investigated the effect of fenoterol-induced constitutive beta(2)-adrenoceptor activity on muscarinic receptor agonist- and histamine-induced bovine tracheal smooth muscle contractions. Bovine tracheal smooth muscle strips were incubated with 10 microM fenoterol or vehicle for various periods of time (5, 30 min, 18 h) at 37 degrees C. After extensive washout (3 h, 37 degrees C), isometric contractions were measured to the full muscarinic receptor agonist methacholine, the partial muscarinic receptor agonist 4-(m-chlorophenyl-carbamoyloxy)-2-butynyltrimethylammonium (McN-A-343) and histamine. Fenoterol treatment significantly reduced the sensitivity (pEC(50)) to methacholine in a time-dependent manner, without affecting maximal contraction (E(max)). Fenoterol treatment similarly reduced the pEC(50) of McN-A-343 and histamine; however, E(max) values were also reduced, to approximately 70% of control after 18-h treatment. The inverse agonist timolol, having no effect on control preparations, consistently restored the reduced pEC(50) and E(max) values of the contractile agonists. Remarkably, in the presence of timolol the pEC(50) values of McN-A-343 and histamine in fenoterol-treated airways were significantly enhanced compared to controls. In conclusion, fenoterol-induced constitutive beta(2)-adrenoceptor activity reduces muscarinic receptor agonist- and histamine induced contractions of bovine tracheal smooth muscle, which can be reversed by the inverse agonist timolol. Moreover, after beta(2)-adrenoceptor agonist treatment, inverse agonism by beta-adrenoceptor antagonists may cause enhanced airway reactivity to contractile mediators. PMID- 11730730 TI - Central antiemetic effects of AS-8112, a dopamine D2, D3, and 5-HT(3) receptor antagonist, in ferrets. AB - The involvement of a central mechanism in the antiemetic effect of AS-8112 ((R)-5 bromo-N-(1-ethyl-4-methylhexahydro-1H-1,4-diazepin-6-yl)-2-methoxy-6-methylamino 3-pyridinecarboxamide x 2 fumarate), a novel and potent dopamine D2, D3, and 5 HT(3) receptor antagonist, was investigated in ferrets. Intracerebroventricularly administered AS-8112 dose dependently inhibited R(+)-7-OH-DPAT (R(+)-7-hydroxy-2 (N,N-di-n-propylamino) tetraline)-induced emesis (ID(50); 0.11 microg/kg, i.c.v.). In addition, AS-8112 (10 microg/kg, i.c.v.) significantly inhibited emesis induced by cisplatin. Ondansetron (10 microg/kg, i.c.v.) also inhibited cisplatin-induced emesis, but did not inhibit R(+)-7-OH-DPAT-induced emesis. S(-) eticlopride (10 microg/kg, i.c.v.) did not inhibit emesis induced by cisplatin. However, racemic CP-99,994 ((+/-)-(2S, 3S)-3-(2-methoxybenzylamino)-2 phenylpiperidine) (10 microg/kg, i.c.v.) inhibited both cisplatin- and R(+)-7-OH DPAT-induced emesis. These results suggest that the antiemetic effects of AS-8112 are centrally mediated via dopamine D3 and 5-HT(3) receptors in ferrets. PMID- 11730731 TI - Beclomethasone, budesonide and fluticasone propionate inhibit human neutrophil apoptosis. AB - Inhaled glucocorticoids are widely used to treat chronic obstructive pulmonary disease without much evidence of efficiency in this disease where neutrophils may contribute to the pathophysiology. This prompted us to test the effects of several currently used inhaled and systemic glucocorticoids on human neutrophil apoptosis. Beclomethasone, budesonide, dexamethasone, fluticasone propionate, hydrocortisone and prednisolone inhibited apoptosis in a concentration-dependent manner as assessed by flow cytometric analysis, annexin-V binding and morphological analysis. The maximal inhibition of apoptosis was 50-60%. The order of potency was fluticasone propionate (EC(50) 0.6+/-0.2 nM) approximately equal to budesonide (EC(50) 0.8+/-0.2 nM)> dexamethasone approximately equal to prednisolone approximately equal to beclomethasone approximately equal to hydrocortisone. The inhibitory effects of glucocorticoids were reversed by mifepristone. Moreover, glucocorticoids slightly enhanced the inhibitory effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on neutrophil apoptosis. The present data suggests that budesonide and fluticasone propionate prolong human neutrophil survival by inhibiting apoptosis at clinically relevant drug concentrations via an effect on glucocorticoid receptor. PMID- 11730732 TI - Leukemia inhibitory factor in human reproduction. AB - OBJECTIVE: To describe the clinical findings, expressions, interactions, and clinical implications of leukemia inhibitory factor (LIF) in human reproduction. DESIGN: Review of published articles. SETTING: Clinical development unit of biotechnology company. INTERVENTION(S): None. RESULT(S): In the endometrium, LIF is expressed in a menstrual cycle-dependent manner, with the highest level occurring at the time of implantation. LIF is also detected in uterine flushing, and its level is significantly lower in women with unexplained infertility. Likewise, endometrial explants derived from women with unexplained infertility showed reduced levels of LIF secretion. Binding of LIF to LIF receptor and gp130 activates signal transduction pathways. LIF receptor is expressed in endometrium, oocytes, and blastocysts. Cytotrophoblasts cultured in the presence of LIF differentiate toward an anchoring extravillous phenotype. CONCLUSION(S): On the basis of reports gathered from animal and human studies, LIF appears to play an important role in implantation and in the establishment of pregnancy. PMID- 11730733 TI - Multiple novel roles of luteinizing hormone. PMID- 11730734 TI - Long-term estrogen replacement is associated with improved nonverbal memory and attentional measures in postmenopausal women. AB - OBJECTIVE: To determine the cognitive domains improved or preserved by long-term hormone replacement therapy (HRT). DESIGN: A comprehensive neuropsychological test battery was administered to healthy postmenopausal women who had been treated or not treated with long-term HRT without interruption since menopause. SETTING: Women were recruited by advertisement from a university town and surrounding areas. PATIENT(S): Women 60 years or older were studied who were treated (n = 16) or not treated (n = 13) with HRT. INTERVENTION(S): Neuropsychological testing included tests of memory, verbal fluency, executive functions, attention and concentration, and psychomotor function. Tests of intellectual function, depressive symptoms, and emotional functioning assessed general functions and comparability of the groups. MAIN OUTCOME MEASURE(S): Neuropsychological testing scores were compared between groups. RESULT(S): No statistically significant differences between the groups were found for general demographic, intellectual, and psychological measures. Scores from both the Weschler Memory Scale Visual Reproduction (delayed recall) and the Digit Vigilance Test (attention) showed statistically significant better performance and fewer errors in the group of women on HRT. CONCLUSION(S): Long-term postmenopausal HRT is associated with higher scores in tests of nonverbal memory and attention. PMID- 11730735 TI - Absorption and preferential vagina-to-uterus distribution after vaginal administration of (99m)Tc-pertechnetate in postmenopausal women. AB - OBJECTIVE: To investigate in vivo and in humans the mechanisms and kinetics of vagina-to-uterus distribution. DESIGN: Controlled clinical study. SETTING: Volunteers in an academic research environment. PATIENT(S): Six postmenopausal women undergoing transabdominal hysterectomy were selected. INTERVENTION(S): Women received 0.2 mL of (99m)Tc-pertechnetate vaginally. In three patients the cervical canal was previously sealed by means of surgical glue. Six postmenopausal women who had received (99m)Tc-pertechnetate intravenously for a thyroid scintigraphy were considered as a control. MAIN OUTCOME MEASURE(S): Radioactivity was assessed every 30 minutes for 6 hours in the pelvis and in body regions where (99m)Tc-pertechnetate normally accumulates (thyroid, salivary glands, and stomach). RESULT(S): Uterine activity appeared after 60 minutes and peaked between 120 to 210 minutes. These same times were observed in the patients who had a sealed cervix. Thyroid uptake appeared after 180 minutes and peaked between 210 and 330 minutes. Uterine uptake did not occur in any of the intravenous patients; their thyroid uptake was rapid, appearing after 30 minutes. CONCLUSION(S): Preferential vagina-to-uterus distribution, at least in postmenopausal women, is not simply due an intracanalicular passage but is mediated by absorption of substances and probably by a countercurrent transfer mechanism. PMID- 11730736 TI - Male partner screening before in vitro fertilization: preselecting patients who require intracytoplasmic sperm injection with the sperm penetration assay. AB - OBJECTIVES: To determine the diagnostic accuracy of the sperm penetration assay (SPA) and standard semen parameters for subsequent fertilization in in vitro fertilization-embryo transfer (IVF-ET). DESIGN: Prospective study. SETTING: Andrology Laboratory, and university research laboratory. PATIENTS: Two hundred sixteen couples undergoing male-partner screening before IVF-ET (265 cycles). INTERVENTION(S): Male-partner screening (semen analyses [SA] and SPA), standard IVF-ET procedures, follow-up of fertilization in IVF-ET. MAIN OUTCOME MEASURE(S): Diagnostic accuracy of SA and SPA for prediction of fertilization in IVF-ET. RESULT(S): The SPA predicted IVF fertilization with high negative (84%) and positive (98%) predictive rates, and correct prediction in 88% of cycles. In contrast, sperm concentration, motility, morphology, and complete SA showed poor diagnostic accuracy, with correct prediction of IVF fertilization in 64%, 65%, 45%, and 68% of cycles, respectively. CONCLUSION(S): Very low sperm concentration and/or motility were good predictors of poor IVF fertilization, however, low to normal semen parameters were not predictive of successful IVF fertilization. The SPA is a useful screening tool that predicts IVF fertilization with high diagnostic accuracy. The SPA may be useful to discriminate between those couples with a high probability of normal fertilization in IVF and those with a low probability of normal fertilization that may benefit from assisted fertilization by intracytoplasmic sperm injection (ICSI). PMID- 11730737 TI - Effect of paternal age on human sperm chromosomes. AB - OBJECTIVE: To investigate whether increased age alters the frequency and type of chromosomal anomalies in human spermatozoa. DESIGN: Semen specimens were collected from donors via masturbation; cytogenetic studies were performed on sperm chromosomes after heterologous (human-hamster) in vitro fertilization. SETTING: Cytogenetics Laboratory, Genetics Department, Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, Brazil. PATIENT(S): Seven men ages 59-74 (older group) and five men ages 23-39 (control group). MAIN OUTCOME MEASURE(S): Frequency and types of chromosomal anomalies in older and control group donors. RESULT(S): The frequency of numerical and structural aberrations (acentric fragments and complex radial figures) was significantly greater in chromosomes of older donors when compared with those of the control group. CONCLUSION(S): The higher frequency of sperm chromosome aberrations in older men was mainly a result of increased nondisjunction, acentric fragments, and complex radial figures. PMID- 11730738 TI - Serum inhibin B may be a reliable marker of the presence of testicular spermatozoa in patients with nonobstructive azoospermia. AB - OBJECTIVE: To establish the predictive value of serum inhibin B levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia, compared with the traditional serum FSH marker. DESIGN: Prospective study. SETTING: Private high-complexity reproductive center with university affiliation. PATIENT(S): Seventy-eight patients with nonobstructive azoospermia, 15 patients with obstructive azoospermia, and 10 fertile volunteers. INTERVENTION(S): Blood samples, testicular sperm extraction, percutaneous epididymal sperm aspiration, and semen collection. MAIN OUTCOME MEASURE(S): Serum levels of inhibin B and FSH and presence of spermatozoa on TESE, PESA, or regular semen analysis. RESULT(S): Patients with nonobstructive azoospermia has significantly higher levels of serum FSH and significantly lower levels of inhibin B. Mean inhibin B serum levels were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa were found (89.31 +/- 73.24 pg/mL vs. 19.23 +/- 22.34 pg/mL), but mean FSH serum levels did not have similar predictive power (21.37 +/- 12.92 IU/mL vs. 19.27 +/- 10.28 IU/mL). The cut-off level of inhibin B separating both groups, as determined by the receiver-operating characteristic curves, was >53 pg/mL. CONCLUSION(S): Serum inhibin B level seems to be more accurate than serum FSH level in prediction of the presence of testicular spermatozoa in patients with nonobstructive azoospermia. PMID- 11730739 TI - Delayed timing of intrauterine insemination results in a significantly improved pregnancy rate in female partners of quadriplegic men. AB - OBJECTIVE: To review pregnancy rates obtained with three protocols used during development of a successful therapy for infertility in couples in which the male partner had spinal cord injury. DESIGN: Retrospective chart review. SETTING: Private infertility center. PATIENT(S): Eleven quadriplegic men and their spouses undergoing intrauterine insemination. INTERVENTION(S): Protocol 1: Intrauterine insemination was performed 24 hours after the LH surge was detected in unstimulated cycles. Sperm were prepared by standard sperm washing. Protocol 2: Female partners were stimulated with clomiphene citrate and hCG. Sperm were inseminated 32-34 hours after hCG injection. Sperm preparation was by serum swim up or density gradient preparation. Protocol 3: Identical to protocol 2, except the insemination was delayed to 38-40 hours after hCG injection. MAIN OUTCOME MEASURE(S): Pregnancy rates. RESULT(S): Five patients were enrolled into protocol 1 and underwent a total of 19 inseminations with no subsequent pregnancies. They then underwent protocol 2, but no pregnancies resulted from inseminations. Four of the original couples, along with six additional couples, underwent insemination in protocol 3. A total of 19 inseminations were performed, and 6 of the 10 patients (60%) became pregnant. The success of insemination at 38-40 hours after hCG administration was significantly better than that of the initial two protocols (P<.05). No differences were observed in sperm quality between protocol 2 and protocol 3. Overall, 73% (8 of 11) of the patients became pregnant. CONCLUSION(S): Intrauterine insemination 38-40 hours after the hCG injection results in an improved chance of pregnancy. These results indicate that many couples with spinal cord injury-associated male infertility can be treated with intrauterine insemination of sperm treated by serum swim-up, with a high probability of success. PMID- 11730740 TI - Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility. AB - OBJECTIVE: To examine the expression of the heat shock protein hsp70-2, and the possible relationship with the pathogenesis of male infertility. DESIGN: Prospective study. SETTING: Reproductive testing laboratory in a university hospital. PATIENT(S): Men undergoing testicular biopsy during an investigation of subfertility. INTERVENTION(S): Testicular tissues were obtained from biopsies of men undergoing infertility evaluation and subdivided into three groups: normal testes, maturational arrest and Sertoli cell-only syndrome. Immunostaining and Western blotting techniques determined expression of the heat shock protein hsp70 2 MAIN OUTCOME MEASURE(S): Expression of the heat shock protein hsp70-2 in the testes. RESULT(S): The experimental data demonstrated that the heat shock protein hsp70-2 was expressed in the normal and maturation arrest testicular specimens. The heat shock protein hsp70-2 was strongly present in the cytoplasm of spermatocytes and spermatides in the adluminal compartment of the seminiferous epithelium in normal testis. However, maturation arrest testis tissue demonstrated light staining in spermatocytes and spermatides, and Sertoli-only specimens demonstrated no staining for the heat shock protein hsp70-2. The Western blotting data showed a 70-kDa heat shock protein in the normal and maturation arrest testicular tissues, but not in the Sertoli-only tissues. CONCLUSIONS: These results suggest that the heat shock protein hsp70-2 is expressed in spermatocytes and spermatides in normal and maturation arrest tissues. However, the expression of the heat shock protein hsp70-2 was low in maturation arrest, and no heat shock protein hsp70-2 was demonstrated in Sertoli only specimens. Therefore the decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility. PMID- 11730741 TI - Delaying the initiation of progesterone supplementation results in decreased pregnancy rates after in vitro fertilization: a randomized, prospective study. AB - OBJECTIVE: To compare IVF outcome between two protocols for luteal phase supplementation, one beginning on day 3 after oocyte retrieval and the other beginning on day 6 after retrieval. DESIGN: Prospective, randomized study. SETTING: University-based assisted reproductive technology center. PATIENT(S): One hundred twenty-six consecutive patients undergoing IVF between January and July 2000. INTERVENTION(S): Patients were randomized to begin luteal phase support using vaginal progesterone beginning either on day 3 after oocyte retrieval or on day 6 after oocyte retrieval. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates and implantation rates. RESULT(S): All patients randomized underwent transfer. There were no differences in age, oocytes retrieved, or embryos transferred between the two groups. Those patients receiving luteal phase support with progesterone beginning on day 6 after retrieval had a significantly lower clinical pregnancy rate per transfer compared with those beginning support on day 3 after retrieval (44.8% vs. 61.0%, respectively). This difference in pregnancy rates was greater in those patients undergoing a luteal gonadotropin releasing hormone (GnRH) agonist down-regulation protocol (47.5% vs. 71.4%, day 6 vs. day 3, respectively). Beginning support on day 6 also significantly decreased implantation rates in the GnRH agonist group (21.0% vs. 34.0%, day 6 vs. day 3, respectively). CONCLUSION(S): Pregnancy rates are significantly decreased by initiating luteal-phase progesterone supplementation on day 6 after oocyte retrieval during in vitro fertilization cycles. PMID- 11730743 TI - Assessment of early cleaving in vitro fertilized human embryos at the 2-cell stage before transfer improves embryo selection. AB - OBJECTIVE: To determine the most viable embryos for transfer. DESIGN: Study 1: Preselection of early-cleaving 2-cell embryos for transfer. Study 2: Alternating weeks during which preselection was performed and not performed. SETTING: ART program, Birmingham Women's Hospital, Birmingham, United Kingdom. PATIENT(S): Patients undergoing IVF or ICSI cycles with transfer on day 2. INTERVENTION(S): Culture of all fertilized embryos. MAIN OUTCOME MEASURE(S): Number of fertilized embryos cleaving to the 2-cell stage on day 1, embryo quality, implantation rates, and pregnancy rates. RESULT(S): Patients with early-cleaving 2-cell embryos had significantly higher pregnancy and implantation rates (45 of 100 [45.0%] and 58 of 219 [25.5%], respectively) than did patients without early cleaving 2-cell embryos (31 of 130 [23.8%] and 43 of 290 [14.8%], respectively). In weeks during which preselection was used, the overall pregnancy and implantation rates of the clinic improved. CONCLUSION(S): The presence of early cleaving 2-cell embryos improves a patient's chance of achieving pregnancy. Use of more stringent embryo selection criteria can improve overall pregnancy rates. PMID- 11730742 TI - A randomized study comparing Crinone 8% and intramuscular progesterone supplementation in in vitro fertilization-embryo transfer cycles. AB - OBJECTIVE: To compare the efficacy of Crinone 8% intravaginal progesterone gel vs. IM progesterone for luteal phase and early pregnancy support after IVF-ET. DESIGN: Randomized, open-label study. SETTING: Academic medical center. PATIENT(S): Two hundred and one women undergoing IVF-ET. INTERVENTION(S): Women were randomized to supplementation with Crinone 8% (90 mg once daily) or IM progesterone (50 mg once daily) beginning the day after oocyte retrieval. MAIN OUTCOME MEASURE(S): Pregnancy, embryo implantation, and live birth rates. RESULT(S): The women randomized to luteal phase supplementation with IM progesterone had significantly higher clinical pregnancy (48.5% vs. 30.4%; odds ratio [OR], 2.16; 95% confidence interval [CI], 1.21, 3.87), embryo implantation (24.1% vs. 17.5%; OR, 1.89; 95% CI, 1.08, 3.30), and live birth rates (39.4% vs. 24.5%; OR, 2.00; 95% CI, 1.10, 3.70) than women randomized to Crinone 8%. CONCLUSION(S): In women undergoing IVF-ET, once-a-day progesterone supplementation with Crinone 8%, beginning the day after oocyte retrieval, resulted in significantly lower embryo implantation, clinical pregnancy, and live birth rates compared with women supplemented with IM progesterone. PMID- 11730744 TI - Quantitative grading of a human blastocyst: optimal inner cell mass size and shape. AB - OBJECTIVE: To investigate the predictive value of quantitative measurements of blastocyst morphology on subsequent implantation rates after transfer. DESIGN: Prospective observational study. SETTING: Private assisted reproductive technology center. PATIENT(S): One hundred seventy-four IVF patients receiving transfers of expanded blastocyst-stage embryos on day 5 (n = 112) or day 6 (n = 62) after oocyte retrieval. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Blastocyst diameter, number of trophectoderm cells, inner cell mass (ICM) size, ICM shape, and implantation and pregnancy rates. RESULT(S): Blastocyst diameter and trophectoderm cell numbers were unrelated to implantation rates. Day 5 expanded blastocysts with ICMs of >4,500 microm(2) implanted at a higher rate than did those with smaller ICMs (55% vs. 31%). Day 5 expanded blastocysts with slightly oval ICMs implanted at a higher rate (58%) compared with those with either rounder ICMs (7%) or more elongated ICMs (33%). Implantation rates were highest (71%) for embryos with both optimal ICM size and shape. Pregnancy rates were higher for day 5 transfers of optimally shaped ICMs compared with day 5 transfers of optimally sized ICMs. CONCLUSION(S): Quantitative measurements of the inner cell mass are highly indicative of blastocyst implantation potential. Blastocysts with relatively large and/or slightly oval ICMs are more likely to implant than other blastocysts. PMID- 11730746 TI - Noninvasive assessment of human embryo nutrient consumption as a measure of developmental potential. AB - OBJECTIVE: To determine the relationship between blastocyst development and morphology and embryo metabolism. DESIGN: Noninvasive assessment of carbohydrate uptake and ammonium production by individual embryos. SETTING: Private assisted reproductive technology unit. PATIENT(S): Patients donated, with consent, cryopreserved pronucleate embryos and noncryopreserved blastocysts. INTERVENTION(S): Culture of 60 thawed pronucleate embryos in sequential media to the blastocyst stage with concomitant noninvasive analysis of embryo metabolism and analysis of 13 blastocysts from noncryopreserved embryos. MAIN OUTCOME MEASURE(S): Pyruvate and glucose consumption as well as blastocyst formation and quality. RESULT(S): Pyruvate and glucose uptakes on day 4 were significantly higher by embryos that went on to form blastocysts than by embryos that failed to develop to the blastocyst stage. Glucose uptakes were greatest in those blastocysts of highest grade, whereas pyruvate uptakes were similar irrespective of blastocyst grade, indicating that glucose is the more important nutrient for the human blastocyst. Among blastocysts of the same grade from the same patient, there was considerable spread of glucose consumption, indicating that glucose consumption may be of use in identifying blastocysts for transfer. Ammonium production by individual embryos was also measured, reflecting amino acid transamination and use by the human embryo. CONCLUSION(S): The ability to identify in culture the embryo with the highest developmental potential will facilitate the move to single-embryo transfers. PMID- 11730745 TI - Measurement of endometrial tissue blood flow: a novel way to assess uterine receptivity for implantation. AB - OBJECTIVE: To assess endometrial receptivity in terms of endometrial tissue blood flow (ETBF) measured hysterofiberscopically by laser blood-flowmetry, and to examine the technique's effectiveness in an in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) program. DESIGN: A prospective clinical study. SETTING(S): IVF program in a university hospital. PATIENT(S): A total of 75 infertile women with normal menstrual cycles undergoing IVF/ICSI. INTERVENTION(S): ETBF, conventional ultrasonographic, endocrinologic, and histologic parameters for receptivity and immunoreactivity for vascular endothelial growth factor (VEGF) in endometrium were assessed between days 4 and 6 of the luteal phase in a spontaneous menstrual cycle. Then all patients underwent IVF/ICSI. MAIN OUTCOME MEASURE(S): Achievement of clinical pregnancy by IVF/ICSI. RESULT(S): ETBF, VEGF expression, and the number of embryos were significantly higher in the women who became pregnant than in those who did not. By stepwise multiple logistic regression, significant predictors of pregnancy were the number of embryos and ETBF but not conventional receptivity markers. The rate of pregnancy was significantly higher in women with ETBF values of at least 29 mL/min per 100 grams of tissue than in women with lower values (42 vs. 15% in 36 and 39 women, respectively). ETBF was significantly greater in morphologically normal than abnormal uteri. In normal uteri, ETBF was greatest in the fundus. Correspondingly, in normal uteri 85% of gestational sacs were implanted in the fundus. CONCLUSION(S): ETBF is superior to conventional parameters for determining endometrial receptivity for implantation. PMID- 11730747 TI - Gender bias in the disposition of frozen embryos. AB - OBJECTIVE: To examine the gender differences found among couples when choosing the disposition of their frozen embryos. DESIGN: Retrospective chart review. SETTING: University affiliated in vitro fertilization (IVF) center. PATIENTS: Couples undergoing their first cycle of assisted reproductive technology (ART). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Choice of disposition of gametes and embryos. RESULT(S): Gender bias is found when couples choose the disposition of their frozen embryos, but not when they choose the disposition of their gametes. CONCLUSION(S): Gender bias was found in couples who made decisions regarding the disposition of their frozen embryos. PMID- 11730748 TI - Differentiating clinical profiles: predicting good responders, poor responders, and hyperresponders. AB - OBJECTIVE: To describe the different clinical and laboratory diagnostic methods (basal and dynamic tests) available to identify poor-, good-, and high-responder patients undergoing treatment with in vitro fertilization (IVF). DESIGN: Analytical review. SETTING: IVF program. PATIENTS: Women in different age groups undergoing IVF treatment. INTERVENTION(S): Assessment of clinical and laboratory parameters and correlation with outcomes. MAIN OUTCOME MEASURE(S): Response to ovarian stimulation and success of the assisted reproductive technologies (ART). RESULT(S): Age, basal follicle-stimulating hormone (FSH), estradiol and inhibin-B levels, and dynamic testing serve to predict individual response to ovarian stimulation for ART. CONCLUSION(S): Markers of ovarian reserve (day 3 FSH, inhibin B and E(2)) are particularly predictive and useful in guiding the choice of the optimal protocol for ART. However, no tests are absolutely predictive of a successful outcome. For the younger individual, and for the patient at risk of developing ovarian hyperstimulation syndrome (OHSS), assessment and identification of clinical and laboratory parameters predictive of a high response to ovarian stimulation should guide the clinician in choosing an appropriate stimulation protocol, thus attenuating the risk of OHSS. PMID- 11730749 TI - Prophylactic methotrexate after linear salpingostomy: a decision analysis. AB - OBJECTIVE: To compare two strategies for managing women after linear salpingostomy for treatment of tubal pregnancy: observation and prophylactic methotrexate. DESIGN: Decision analysis. SETTING: Outpatient tertiary-care center. PATIENT(S): One thousand hypothetical women treated with a linear salpingostomy for ectopic pregnancy. INTERVENTION(S): Observation after salpingostomy and treatment of persistent ectopic pregnancy with a single dose of methotrexate (current standard of care) versus treatment with prophylactic methotrexate at the time of salpingostomy. MAIN OUTCOME MEASURE(S): Number of ruptured ectopic pregnancies, surgical procedures, complications, and cost for each group (observation vs. prophylaxis). RESULT(S): Prophylactic methotrexate results in fewer cases of tubal rupture (0.4% vs. 3.7%) and fewer procedures (1.9% vs. 4.7%) at a lower cost ($67.55 less/patient) compared with observation alone. Methotrexate-associated complications occur more frequently with prophylaxis (5.5% vs. 0.8%). Certain conditions change which strategy is preferable. Observation is the best strategy when the persistent ectopic pregnancy rate is <9%, the success of prophylaxis is <95%, the complication rate associated with methotrexate is >18%, or the rupture rate of persistent ectopic pregnancies is <7.3%. CONCLUSION(S): Prophylactic methotrexate at the time of linear salpingostomy for the treatment of ectopic pregnancy is preferable to observation as long as certain conditions exist. PMID- 11730750 TI - Noncompliance among a group of women using a novel method of contraception. AB - OBJECTIVE: To compare the incidence of noncompliance measured objectively by a home use fertility monitor with the traditional self-reported incidence of compliance in a study of a new method of contraception. DESIGN: Prospective cohort study. SETTING: A large family planning clinic in Edinburgh. PATIENT(S): Thirty-two healthy women who took part in a trial assessing the efficacy of a novel method of contraception involving accurately timed administration of a single dose of mifepristone. INTERVENTION(S): Mifepristone was administered orally and a blood sample was collected on the same day. MAIN OUTCOME MEASURE(S): Percentage of missed tests detected by the monitor against the self-reported percentage during the critical period. RESULT(S): Women failed to perform 24.2% (95% confidence interval, 16.5-31.5) of the tests in the 162 cycles analyzed. They missed tests at an absolutely vital time for contraceptive efficacy in 42% of cycles according to the monitor while admitting to missing tests in 14.8%. Poor compliance was associated with younger women, those who discontinued the study before completion, and cycles in which women were not relying on the contraceptive method. CONCLUSION(S): The use of microelectronic monitoring systems may improve our understanding of the extent of patient noncompliance, providing objective information that no other monitoring technique can produce. This understanding provides the opportunity to make the optimum use of potentially effective treatments while validating research evidence. PMID- 11730751 TI - Low-penetrance genes are associated with increased susceptibility to endometriosis. AB - OBJECTIVE: To investigate whether genetic polymorphisms of CYP1A1, GSTM1, and GSTT1 are associated with endometriosis. DESIGN: Genetic polymorphism analysis. SETTING: University department. PATIENT(S): A family with four women in two generations who had endometriosis and one member with suspected endometriosis in the third generation were compared with a group of fertile women. INTERVENTION(S): Laparoscopic examination. MAIN OUTCOME MEASURE(S): Blood specimens were obtained from fertile females and available affected female family members. Multiplex polymerase chain reaction (PCR) and restriction fragment length polymorphism PCR was done to determine each participant's genotype. RESULT(S): All affected family members had genotype CYP1A1 wt/m1 and GSTM1 null deletion. The frequency of this genotype in 54 fertile women was 13%. A 17-year old family member with suspected endometriosis had the same genotype. One affected member was also a carrier of a GSTT1 null deletion. This combination was not found in any of the fertile participants. The most frequent genotypes in the sample were CYP1A1 wt/wt, with GSTM1 null deletion and at least one functional allele of GSTT1, and CYP1A1 wt/wt, with at least one functional allele of GSTM1 and GSTT1 (33% and 31%, respectively). CONCLUSION(S): The combination of CYP1A1 m1 polymorphism and GSTM1 null deletion is closely associated with penetration of the endometriosis phenotype, whereas GSTT1 null deletion may add to the penetration of this trait. PMID- 11730752 TI - Matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP in peritoneal fluids and sera and correlation with peritoneal adhesions. AB - OBJECTIVE: To assess the presence of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in peritoneal fluid and serum of subjects with and without adhesions. DESIGN: Cross-sectional study. SETTING: Academic research centers. PATIENT(S): Sixty-three patients who underwent abdominal/pelvic surgery. INTERVENTION(S): MMP-1, TIMP-1, and MMP-1-TIMP-1 complex content. MAIN OUTCOME MEASURE(S): ELISA. RESULT(S): Peritoneal fluids (PF) and sera of subjects with and without peritoneal adhesions contain MMP-1, TIMP-1, and MMP-1-TIMP-1 complex at varying levels with 10- to 100-fold higher TIMP-1 than MMP-1. Compared with serum, PF contains a lower level of MMP-1 in subjects with mild adhesions and without adhesions, higher TIMP-1 in subjects with extensive adhesions, and lower MMP-1-TIMP-1 complex in subjects with moderate adhesions. However, the serum and PF content of MMP-1, TIMP-1, and MMP-1-TIMP-1 complex was not statistically different among subjects with or without adhesions, with the exception of TIMP-1 in PF of subjects with extensive adhesions. MMP1-TIMP-1 ratio indicates that a major portion of MMP-1 is in complex with TIMP-1. There was no age- or gender dependent difference in MMP-1 and TIMP-1 content in serum or PF. CONCLUSION(S): Despite differences in MMP-1 and TIMP-1 levels in serum and PF of subjects with extensive and moderate adhesions, there is no correlation between MMP-1 and TIMP 1, with the exception of higher TIMP-1 in PF of subjects with extensive adhesions. PMID- 11730753 TI - Expression of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP in serosal tissue of intraperitoneal organs and adhesions. AB - OBJECTIVE: To compare expression of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in serosal tissue of intraperitoneal organs and adhesions. DESIGN: Prospective and cross-sectional study. SETTING: Academic research centers. PATIENT(S): Patients undergoing abdominal or pelvic surgery. INTERVENTION(S): MMP-1 and TIMP-1 expression. MAIN OUTCOME MEASURE(S): Expression of messenger ribonucleic acid (mRNA) and protein was measured by using quantitative reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay. RESULT(S): Serosal tissue of intraperitoneal organs and adhesions express MMP-1 and TIMP-1 mRNA and protein at levels that are consistently varied with 10- to 10,000-fold and 2- to 10-fold higher TIMP, mRNA and protein, respectively. Parietal peritoneum, fallopian tubes and ovaries express higher MMP-1 mRNA levels compared with uterus and adhesions; the lowest expression is found in small and large bowels, subcutaneous tissue. and omentum. Expression of TIMP-1 mRNA was less variable; the highest level was found in the uterus and the lowest in subcutaneous tissue and small bowels. There was less variability in MMP-1 and TIMP-1 protein content than mRNA expression; ovaries and adhesions contained the highest MMP-1 and TIMP-1 levels, respectively, and peritoneum contained the lowest. The MMP-1 and TIMP-1 content and ratios further indicate limited MMP-1 proteolytic activity. Although tissues from premenopausal women express more MMP-1 and TIMP-1, expression did not differ by sex or age. CONCLUSION(S): Because MMP-1 and TIMP-1 expression varies consistently among the serosal tissues of peritoneal organs and adhesions, and because tissue injury alters their expression, site-specific variations in expression of these substances may predispose a particular organ to develop more adhesions. PMID- 11730754 TI - Vascular endothelial growth factor mRNA in eutopic and ectopic endometrium. AB - OBJECTIVE: To evaluate changes in expression levels of vascular endothelial growth factor (VEGF) mRNA in human endometrial explants in a chicken chorioallantoic membrane model of endometriosis. DESIGN: Experimental prospective study. SETTING: University hospital. PATIENT(S): Endometrial biopsy samples were obtained from healthy, ovulating women undergoing elective surgery. INTERVENTION(S): Endometrial fragments were placed on the chicken chorioallantoic membrane and removed for analysis after 0, 24, 48, and 72 hours. MAIN OUTCOME MEASURE(S): Expression of different VEGF mRNA splice variants was tested. Expression of VEGF(165) mRNA was assessed by using competitive polymerase chain reaction and normalized to expression of the housekeeping gene human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. RESULT(S): After 0, 24, 48, and 72 hours of incubation, all grafts expressed VEGF(121), VEGF(145), VEGF(165), and VEGF(189) mRNA. Expression of VEGF(165) mRNA increased up to 10 fold at 24 to 72 hours compared with precultivation values. CONCLUSION(S): Levels of VEGF(165) mRNA in endometrial grafts increase after implantation on chicken chorioallantoic membrane. Hypoxic induction of VEGF mRNA expression in endometrial cell cultures has been reported previously. Induction of VEGF expression might indicate relative hypoxia of the specimen due to insufficient vascularization. Expression of VEGF may assist in vascularization of endometrial explants after retrograde menstruation. PMID- 11730755 TI - Progesterone receptor antagonists Org 31710 and RU 486 increase apoptosis in human periovulatory granulosa cells. AB - OBJECTIVE: To investigate if progesterone receptor (PR)-mediated effects are involved in regulating the susceptibility to apoptosis in LH receptor-stimulated human luteinizing granulosa cells. DESIGN: Laboratory study. SETTING: Goteborg University and an in vitro fertilization laboratory of a university hospital. PATIENT(S): Women undergoing oocyte retrieval for in vitro fertilization after ovulation induction with gonadotropins. INTERVENTION(S): Luteinizing granulosa cells were isolated from follicular aspirates after oocyte removal. The cells were treated with or without RU 486 (1 microM-100 microM), Org 31710 (1 microM 100 microM), progesterone (1 nM-10 microM), dexamethasone (0.5 microM-100 microM), dihydrotestosterone (1 nM-25 microM), RU 486 (10 microM-100 microM) + dexamethasone (50 microM), and picrotoxin (1 microM-100 microM) and were cultured under serum-free conditions. MAIN OUTCOME MEASURE(S): Measurement of caspase-3 activity; detection of internucleosomal DNA fragmentation using gel electrophoresis and fluorospectrophotometry; progesterone analysis of spent medium. RESULT(S): Addition of the PR antagonists RU 486 or Org 31710 in vitro to human luteinizing granulosa cells caused an increase in caspase-3 activity and a dose-dependent increase in internucleosomal DNA fragmentation. No effect on DNA fragmentation was seen after addition of dexamethasone, dihydrotestosterone, or picrotoxin. CONCLUSION(S): Nuclear PR-mediated effects are involved in regulating the susceptibility to apoptosis in LH receptor-stimulated human luteinizing granulosa cells. PMID- 11730756 TI - Effect of angiotensin-converting enzyme inhibitor on renal function in ovarian hyperstimulation syndrome in the rabbit. AB - OBJECTIVE: To investigate renal function and whether captopril prevents alterations in the handling of sodium and water in the ovarian hyperstimulation syndrome (OHSS) in the rabbit. DESIGN: Experimental study SETTING: Physiology laboratory. ANIMAL(S): Six female New Zealand white rabbits were used as controls, and 13 were hyperstimulated with gonadotropins. INTERVENTION(S): Saline or captopril. MAIN OUTCOME MEASURE(S): Renal excretory and hemodynamic variables. RESULT(S): The 3% extracellular volume expansion in OHSS animals induced a significant elevation in mean arterial pressure by 27%, although increments in natriuresis and diuresis were similar to those observed in controls. The OHSS group had impaired pressure-natriuresis sensitivity compared with controls (0.36 +/- 0.07 microEq/min/g of Na excreted per mm Hg vs. 1.74 +/- 0.45 microEq/min/g of Na excreted per mm Hg; P<.05. Captopril significantly reduced mean arterial pressure (P<.05) and shifted the pressure-natriuresis response to the left by 0.85 +/- 0.17 microEq/min/g of Na excreted per mm Hg (P<.05). CONCLUSION(S): In OHSS in the rabbit model, pressure-natriuresis sensitivity is impaired. Angiotensin II may play a significant role in this phenomenon, since angiotensin converting enzyme inhibition normalized the pressure-natriuresis relationship. PMID- 11730757 TI - Risk and outcome of bowel injury during transvaginal pelvic endoscopy. AB - OBJECTIVE: To determine the risk and outcome of bowel injury associated with new techniques of transvaginal pelvic endoscopy. DESIGN: A multinational retrospective survey based on confidential, self-reported cases. SETTING: Tertiary referral centers for infertility. PATIENT(S): Infertile patients without obvious pelvic pathology. INTERVENTION(S): Transvaginal hydrolaparoscopy and fertiloscopy. MAIN OUTCOME MEASURE(S): Full-thickness bowel injury. RESULT(S): Thirty-nine responders reported a total of 24 bowel injuries (0.65%) in 3667 procedures. After initial experience, the prevalence of bowel injury was 0.25%. All of the injuries were diagnosed during the procedure. Twenty-two (92%) of the cases were managed without consequences. CONCLUSION(S): The bowel lesion caused by the new techniques of transvaginal pelvic endoscopy tends to be minor and under strict conditions can be treated expectantly. PMID- 11730758 TI - A simple method for fallopian tube sperm perfusion using a blocking device in the treatment of unexplained infertility. AB - OBJECTIVE: To evaluate the efficacy of fallopian sperm perfusion (FSP) using a new method similar to the FAST system in comparison with standard intrauterine insemination (IUI) in patients with unexplained infertility. DESIGN: Prospective, randomized, controlled study. SETTING: Assisted conception service in a University Hospital. PATIENT(S): Women with unexplained infertility undergoing controlled ovarian hyperstimulation (COH). INTERVENTION(S): After hCG administration, patients were randomized to either standard IUI or FSP. The women received the same treatment in the first and all subsequent cycles. A maximum of three cycles was performed. Intrauterine insemination was performed using a standard method, and fallopian sperm perfusion was performed using a commercial device for hysterosalpingography and tubal hydropertubation. MAIN OUTCOME MEASURE(S): Clinical and ongoing pregnancy rates. RESULT(S): A total of 132 cycles was completed: 66 IUI cycles and 66 FSP cycles. In the IUI group, there were 5 ongoing pregnancies, giving a pregnancy rate of 7.6 per cycle and 15.6% per patient; in the FSP group, 14 ongoing pregnancies occurred, giving a pregnancy rate of 21.2% per cycle and 42.4% per patient. The prevalence of multiple pregnancies, miscarriages and ectopic pregnancies was similar in the two insemination groups. Fallopian sperm perfusion was easy to perform, and no case of sperm reflux was observed. The procedure was well tolerated and no complications were observed. The costs were comparable with standard IUI. CONCLUSION(S): In the treatment of couples with unexplained infertility, the method for fallopian sperm perfusion described yields higher pregnancy rates than IUI, with no significant increase in costs or complications. However, these results need to be confirmed in larger studies before replacing IUI with FSP as standard practice. PMID- 11730759 TI - Electroejaculation and assisted reproductive technologies in the treatment of anejaculatory infertility. AB - OBJECTIVE: To determine the efficacy of electroejaculation in combination with assisted reproductive technology (ART). DESIGN: Case series. SETTING: University fertility program. PATIENT(S): One hundred twenty-one consecutive couples seeking treatment of anejaculatory infertility. INTERVENTION(S): Electroejaculation with IUI, or gamete intrafallopian transfer or IVF. MAIN OUTCOME MEASURE(S): Pregnancy and pregnancy outcome. RESULT(S): Fifty-two couples became pregnant (43%), 39 by IUI alone (32.2%). Cycle fecundity for IUI was 8.7%. No difference in cycle fecundity was seen among ovarian stimulation protocols (clomiphene citrate, 7.6%, hMG, 13.2%, and natural cycle, 11.2%). Pregnancy was unlikely when the inseminated motile sperm count was <4 million. Female management protocol and etiology of anejaculation did not affect results. Patients undergoing IVF had higher cycle fecundity (37.2%) than did those undergoing IUI. The rates of spontaneous abortion and multiple gestations were 23% and 12%, respectively. CONCLUSION(S): Electroejaculation with stepwise application of ART is effective in treating anejaculatory infertility. Intrauterine insemination with the least expensive monitoring protocol should be used for most couples, because use of more expensive monitoring did not improve results. It is cost-effective to bypass IUI and proceed directly to IVF in men who require anesthesia for electroejaculation and in those with a total inseminated motile sperm count < 4 million. PMID- 11730760 TI - Congenital obstructive azoospermia in a man with Marfan syndrome. AB - OBJECTIVE: To present a case of Marfan syndrome and concurrent congenital obstructive azoospermia. DESIGN: Case report. SETTING: Tertiary-care academic hospital. PATIENT(S): A man with Marfan syndrome and obstructive azoospermia. INTERVENTION(S): The patient was evaluated for azoospermia that was proved to be due to congenital absence of large segments of vas deferens and epididymis bilaterally. MAIN OUTCOME MEASURE(S): Evaluation for testicular sperm extraction and ICSI procedure. RESULT(S): The patient is a candidate for testicular sperm extraction and ICSI. CONCLUSION(S): The Marfan syndrome and azoospermia may be associated in sporadic cases. PMID- 11730761 TI - Chromosome 15 aneuploidy in the sperm and conceptus of a sibling with variable familial expression of round-headed sperm syndrome. AB - OBJECTIVE: To characterize rates of chromosome aneuploidy in sperm from three siblings, one of whom had an IVF/ICSI conceptus with trisomy 15. DESIGN: Blind evaluation of the sperm chromosome aneuploidy rates, semen quality, and sperm ultrastructure. SETTING: IVF clinic and university-based andrology research laboratory. PATIENT(S): Three brothers, two of whom underwent infertility evaluation and therapy. MAIN OUTCOME MEASURE(S): Semen from three siblings was coded and blindly evaluated for standard World Health Organization semen quality variables and sperm ultrastructure. Sperm were decondensed and hybridized with fluorescent probes specific for chromosomes X, Y, 13, 15, 18, and 21, then evaluated microscopically to determine the aneuploidy rate for those chromosomes. RESULT(S): Two siblings had increased round-headed morphology on standard morphology evaluation, which was confirmed using electron microscopy. The sperm aneuploidy rate was significantly increased for chromosome 15 in sibling 1, the father of a conceptus with trisomy 15. Aneuploidy rates were also slightly increased for chromosomes X, Y, and 18 in sibling 1. CONCLUSION(S): This is the second report of increased sperm chromosome aneuploidy in infertile patients with round-headed sperm. Although ICSI is successful in treating this syndrome, the risk for aneuploidy of the conceptus may be increased. Other studies have reported an increased incidence of sperm chromosome aneuploidy in some infertile patients, but this is the first report of aneuploidy in both the sperm and conceptus of a patient undergoing IVF/ICSI. PMID- 11730762 TI - Ambiguous genitalia, 45,X/46,XY mosaic karyotype, and Y chromosome microdeletions in a 17-year-old man. AB - OBJECTIVE: To describe clinical and laboratory features of a patient with 45,X/46,XY mosaic karyotype and Y chromosome microdeletions and to discuss the diagnostic problems in his management. DESIGN: Case report. SETTING: University department. PATIENT(S): A 17-year-old man with ambiguous genitalia, 45,X/46,XY mosaic karyotype, and Y chromosome microdeletions. INTERVENTION(S): Testicular ultrasonography, karyotype, open testicular biopsy, polymerase chain reaction (PCR) screening for cystic fibrosis, PCR screening for Y chromosome microdeletions in peripheral blood and testicular tissue, and reverse transcriptase PCR in testicular tissue for Y chromosome microdeletions. MAIN OUTCOME MEASURE(S): Avoidance of dissemination of testicular cancer. RESULT(S): The patient was referred for bilateral orchiectomy. CONCLUSION(S): 45,X/46,XY mosaic karyotype is associated with a broad spectrum of phenotypes that includes female with Turner syndrome, male with mixed gonadal dysgenesis, male pseudohermaphroditism, and apparently normal male. Microdeletions of the long arm of the Y chromosome may be associated with Y chromosomal instability, leading to formation of 45,X cell lines. 45,X/46,XY males carry an increased risk for gonadal tumors and must be followed closely. PMID- 11730763 TI - Ovarian sex cord tumor with annular tubules in a woman with premature ovarian failure. AB - OBJECTIVE: To report a case of premature ovarian failure (POF) associated with an ovarian sex cord tumor with annular tubules. DESIGN: Case report. SETTING: Reproductive endocrinology unit in a tertiary academic center. PATIENT(S): A 20 year-old woman with POF. INTERVENTION(S): Biopsy of the rudimentary ovary by laparoscopy. MAIN OUTCOME MEASURE(S): Protocol for POF investigation and histological study of the ovarian sample. RESULT(S): An ovarian sex cord tumor with annular tubules was detected in the rudimentary right ovary. CONCLUSION(S): The rare ovarian sex cord tumor with annular tubules, which may be hormonally active, was detected in a case of POF. PMID- 11730764 TI - Laparoscopic subtotal hysterectomy and bilateral adnexectomy in a child with congenital adrenal hyperplasia. AB - OBJECTIVE: To report the results of a subtotal hysterectomy and bilateral adnexectomy in a child with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. DESIGN: Case report. SETTING: Gynecology department, Afula, Israel. PATIENT(S): An 11-year-old child with a female genotype and saltwasting type congenital adrenal hyperplasia who was being raised as a boy. INTERVENTION(S): Laparoscopic subtotal hysterectomy and bilateral salpingo-oophorectomy were performed. MAIN OUTCOME MEASURE(S): Surgical efficiency, operating time, recovery, and aesthetic result. RESULT(S): The procedure was performed without complications in 26 minutes, and the child was dismissed the following day. Three 5-mL abdominal incisions were required. CONCLUSION(S): Laparoscopic subtotal hysterectomy and bilateral adnexectomy can be safely performed in children and is the procedure of choice in the rare cases in which it is indicated. PMID- 11730765 TI - Successful treatment of symptomatic arteriovenous malformation of the uterus using laparoscopic bipolar coagulation of uterine vessels. AB - OBJECTIVE: To report a case of symptomatic arteriovenous malformation (AVM) of the uterus that was successfully treated with laparoscopic bipolar coagulation of uterine vessels. DESIGN: Case report. SETTING: University-affiliated tertiary referral center. PATIENT(S): A 66-year-old woman with symptomatic AVM of the uterus. INTERVENTION(S): Laparoscopic bipolar coagulation of uterine vessels. MAIN OUTCOME MEASURE(S): Clinical symptoms, color Doppler sonographic examination, and pelvic magnetic resonance imaging. RESULT(S): Remarkable shrinkage of the lesion size, obvious decrease in all of the impedance measurements (pulsatility, resistance, velocity indexes [systolic/diastolic]), and freedom from symptoms. CONCLUSION(S): This modality is a new alternative method for the management of patients with symptomatic AVM of the uterus who do not respond to conservative treatment. PMID- 11730766 TI - Cytogenetic investigation of fetuses and infants conceived through intracytoplasmic sperm injection. AB - OBJECTIVE: To determine the incidence of aneuploidy among fetuses and infants conceived through intracytoplasmic sperm injection (ICSI) in our clinic using umbilical cord blood samples. DESIGN: Follow-up study of the cytogenetic outcome of ICSI pregnancies. SETTING: University-based IVF clinic. PATIENT(S): Forty-six couples who underwent ICSI and conceived. INTERVENTION(S): Umbilical cord blood was taken after delivery of the infant for analysis. Samples of chorionic villi and chorion were taken for studies on the spontaneous abortuses. Amniocentesis was performed for couples that chose prenatal diagnosis. MAIN OUTCOME MEASURE(S): The cytogenetic chromosomal status of the pregnancy outcome. RESULT(S): Fifty pregnancies and 55 live births were recorded, with nine spontaneous abortions. Of 43 separate umbilical cord blood samples analyzed, 1 abnormality (2%) was found, 45, XX,+21. Nine births went through prenatal diagnosis alone, with four accepting both forms of analysis-no abnormalities were found. Origin of abnormality was established in two spontaneous abortion cases (45, XO and 45, XY, 21), and the maternal chromosome was lost in both cases. CONCLUSION(S): Using umbilical cord blood obtained after birth, we obtained karyotype results from 78% of the ICSI population in our clinic. Combined with results from five additional cases that underwent prenatal diagnosis but not umbilical cord blood sampling, a chromosomal result was obtained in 87% of our ICSI population. The use of umbilical cord blood for cytogenetic analysis substantially improves the ability to determine rates of chromosomal abnormalities in newborns produced via ICSI clinics. PMID- 11730767 TI - Can Doppler ultrasonographic measurements of the testis predict spermatozoa retrieval in azoospermic patients undergoing testicular biopsy? PMID- 11730768 TI - Pregnancy outcome after unilateral salpingostomy with a contralateral patent oviduct. PMID- 11730769 TI - Management of assisted reproductive technologies in women with cystic fibrosis. PMID- 11730770 TI - First-trimester screening for Down's syndrome in in vitro fertilization pregnancies. PMID- 11730771 TI - A simple headstone or just eliminate the chads? PMID- 11730774 TI - A mathematical model for predicting which embryos to transfer-an illusion of control or a powerful tool? PMID- 11730776 TI - Recurrent abortion-antibodies to factor XII or a decrease in factor XII levels? PMID- 11730777 TI - Recombinant FSH vs. urinary FSH--retrospective study of matched pairs for type of treatment (IVF or ICSI). PMID- 11730781 TI - The potential of artemether for the control of schistosomiasis. AB - Schistosomiasis continues to rank--following malaria--at the second position of the world's parasitic diseases in terms of the extent of endemic areas and the number of infected people. There is yet no vaccine available and the current mainstay of control is chemotherapy with praziquantel used as the drug of choice. In view of concern about the development of tolerance and/or resistance to praziquantel, there is a need for research and development of novel drugs for the prevention and cure of schistosomiasis. Interestingly, derivatives of artemisinin, which are already effectively used in the treatment of malaria, also exhibit antischistosomal properties. Significant advances have been made with artemether, the methyl ether derivative of artemisinin. We review the discovery of the antischistosomal activity of artemether by Chinese scientists two decades ago; the detailed laboratory studies of the susceptibility of, and effect on, the different developmental stages of Schistosoma japonicum, Schistosoma mansoni and Schistosoma haematobium to artemether; the possible mechanism of action and the potential long-term toxicity. Finally, we look at the effect of combined treatment with artemether and praziquantel; and clinical findings thus far obtained from randomised controlled trials with oral artemether for the prevention of patent infections and morbidity. The review intends to create a forum for strategic discussion of how these laboratory and clinical findings could be translated into public health actions. We conclude that artemether--as part of integrated current control measures and adapted to specific socio ecological and epidemiological settings--has considerable potential to significantly reduce the current burden of schistosomiasis in many parts of the world. PMID- 11730782 TI - Haemonchus contortus utilises catalase in defence against exogenous hydrogen peroxide in vitro. AB - The toxicity of activated oxygen species towards adult Haemonchus contortus nematodes was examined in in vitro assays using ingestion of [3H]inulin to assess nematode viability. Both glucose/glucose oxidase (generation of hydrogen peroxide) and xanthine/xanthine oxidase (generation of superoxide anion) systems showed concentration-dependent toxicity to the nematodes. Both adult and larval Haemonchus contortus enzyme preparations showed significant catalase activities. Adult nematodes exposed to aminotriazole for 24 h showed catalase activities reduced to less than 20% of controls. Aminotriazole-treated nematodes exposed to a glucose/glucose oxidase system were significantly more susceptible to the toxic effects of the oxidant-generating system than controls (no aminotriazole pre treatment). The concentration of glucose oxidase required to inhibit feeding by 50% was decreased 33-fold in aminotriazole-treated nematodes compared with controls. The effect of aminotriazole pre-treatment implicates hydrogen peroxide as a significant toxic agent in the glucose/glucose oxidase system. It is apparent that inhibition of Haemonchus contortus catalase increases the susceptibility of the parasite to the toxic effects of hydrogen peroxide, demonstrating a protective role for this enzyme. This suggests that catalase has the potential to play a significant role in the defence of this parasite against hydrogen peroxide produced as part of the respiratory burst of activated phagocytes within the host during its response to nematode infection. PMID- 11730783 TI - Identical digeneans in coral reef fishes from French Polynesia and the Great Barrier Reef (Australia) demonstrated by morphology and molecules. AB - Three coral reef fish species, Zanclus cornutus, Chaetodon vagabundus and Naso lituratus, were collected in French Polynesia and on the Great Barrier Reef, Queensland. These fish species were each infected by one morphologically similar digenean species in both localities; Schistorchis zancli Hanson, 1953 was found in Zanclus cornutus, Preptetos laguncula Bray and Cribb, 1996 in Naso lituratus and Neohypocreadium dorsoporum Machida and Uchida, 1987 in Chaetodon vagabundus. In addition, on the Great Barrier Reef P. laguncula was also found in Naso unicornis and N. dorsoporum in Chaetodon ephippium and Chaetodon flavirostris. Morphometric differences between the species from the two sites were only slight. Sequences from the second internal transcribed spacer of the ribosomal DNA of each worm revealed total homology or negligible divergence between samples from hosts caught in French Polynesia and on the Great Barrier Reef. These results show that across more than 6000 km these digeneans are similar in morphology and genotype. Some species of fishes and molluscs are considered to have distributions that encompass the entire tropical Indo-West Pacific. These findings suggest that at least some of their parasites have similarly broad distributions. PMID- 11730784 TI - Assessment of the epidemiological status of Echinococcus multilocularis in foxes in France using ELISA coprotests on fox faeces collected in the field. AB - The aim of this study was to estimate the relevance of Echinococcus multilocularis coproantigen detection in fox faeces collected in the field to identify different levels of endemicity for Echinococcus multilocularis on a large scale (n x 10 km(2)). Six study sites were selected in a high endemicity area and two study sites in a low endemicity area in eastern France on the basis of landscape composition. Sampling was undertaken in the winters of 1996-97, 1997 98 and 1998-99. At each site, (i) necropsy and intestine examination was undertaken on a sample of shot foxes (total number of foxes, 222), and (ii) fox faeces were collected in the field along road verges, and scored for degradation status (total number of faeces, 625). Fox faeces were also sampled in a control area (n=30) in western France in the summer of 1998. Intestines were examined according to the sedimentation method. Echinococcus multilocularis coproantigens were detected by using two ELISA tests: EM-ELISA and EmA9-ELISA. The necropsy prevalence in high and low endemicity areas was 63.3% and 19.4%, respectively, and the distribution of adult worms in the fox population was highly overdispersed (75.5% of the total biomass was harboured by 11.6% of foxes). Using the two ELISA tests, there was no difference in the detection of E. multilocularis coproantigens in field faeces, regardless of the degradation status. The medians of EM- and EmA9-ELISA OD values of field faeces in high endemicity area were significantly higher than in low endemicity area (P<0.001 for both ELISA). The distribution of EM-ELISA OD values in low endemicity area was significantly higher (P=0.002) than in the control area. Moreover, for the two ELISA, the observed ELISA OD value distributions in high endemicity area, low endemicity area and control area seemed representative of the distribution of adult worms in fox populations. These results indicate that E. multilocularis coproantigen detection in field faeces could serve for large-scale surveillance, as an alternative to necropsy. PMID- 11730785 TI - Interspecific associations among larval helminths in fish. AB - Various processes can generate associations between the larvae of different helminth species in their fish intermediate or paratenic host. We investigated the pairwise associations among larval helminth species in eight different fish populations, using two different coefficients of associations, in order to determine in what situations they are strongest. All helminth species included use the fish studied as either their second intermediate host or their paratenic host, and are acquired by the fish when it ingests an infected first intermediate host. The intensity of infection correlated positively with fish length for most helminth species. Pairs of species which both exhibited positive correlations with fish length tended to be more strongly associated with one another, although this tendency was not pronounced. Similarity in life cycle had a more important influence on pairwise associations. Among the 62 pairwise associations that could be computed, pairs of helminth species that shared both first intermediate hosts and definitive hosts were the most strongly associated, followed by pairs that shared only one other host, and finally by pairs that did not share other hosts. The results suggest that assemblages of larval helminth parasites in fish are not random collections of locally available species, but rather structured packets of larval parasites that travel together along common transmission routes. PMID- 11730786 TI - Relationship between worm burden and male proportion in Schistosoma mansoni experimentally infected rodents and primates. A meta-analytical approach. AB - This paper investigates the relationship between the worm burden and the male proportion of adult Schistosoma mansoni. This relationship was tested using data from the literature on S. mansoni experimentally infected rodents and primates. The analyses were performed within each animal model (rat, mouse, hamster and monkey) and showed positive linear relationships between worm burden and male proportion. An analysis between the four animal models showed the same positive linear relationship. The role of the genetic diversity of male and female schistosomes in this relationship is discussed. PMID- 11730787 TI - Sequence variation in the cytochrome oxidase I, internal transcribed spacer 1, and Ts14 diagnostic antigen sequences of Taenia solium isolates from South and Central America, India, and Asia. AB - We examined the genetic variability in the pig-human tapeworm, Taenia solium, by sequencing the genes for cytochrome oxidase I, internal transcribed spacer 1, and a diagnostic antigen, Ts14, from individual cysts isolated from Peru, Colombia, Mexico, India, China, and the Philippines. For these genes, the rate of nucleotide variation was minimal. Isolates from these countries can be distinguished based on one to eight nucleotide differences in the 396 nucleotide cytochrome oxidase I (COI) sequence. However, all of the 15 isolates from within Peru had identical COI sequences. The Ts14 sequences from India and China were identical and differed from the Peru sequence by three nucleotides in 333. These data indicate that there is minimal genetic variability within the species T. solium. Minimal variability was also seen in the ITS1 sequence, but this variation was observed within the individual. Twenty-two cloned sequences from six isolates sorted into 13 unique sequences. The variability observed within the sequences from individual cysts was as great as the variability between the isolates. PMID- 11730788 TI - Genetic diversity of Schistosoma mansoni within and among individual hosts (Rattus rattus): infrapopulation differentiation at microspatial scale. AB - The distribution of genetic diversity in a local population of the trematode Schistosoma mansoni was determined within and between individual wild rats at a microspatial geographic scale of a standing water transmission site. Using RAPD markers, molecular variance and canonical correspondence analysis were performed to test the significance of genetic differentiation between infrapopulations. Of total gene diversity, 8 and 11% was partitioned between hosts trapped at few metres distance from each other. Significant temporal differentiation (2%) was also detected among schistosomes sampled at 6 month intervals with more infrapopulation pairs differentiated during the dry season of parasite transmission than during the rainy season (45 and 12%, respectively). A combination of factors such as restricted displacement of rats, patchy spatial aggregation of infected snails and limited cercarial dispersion in standing water are likely to promote the genetic differentiation observed between infrapopulations at this microgeographic scale. PMID- 11730789 TI - Molecular characterisation and expression of two venom allergen-like protein genes in Heterodera glycines. AB - Secretory proteins encoded by genes expressed in the oesophageal gland cells of plant-parasitic nematodes have key roles in nematode parasitism of plants. Two venom allergen-like protein cDNAs (designated hg-vap-1 and hg-vap-2)were isolated from Heterodera glycines gland cell cDNA libraries. Both cDNAs hybridised to genomic DNA of H. glycines in Southern blots. The hg-vap-1 cDNA contained an open reading frame encoding 215 amino acids with the first 25 amino acids being a putative secretion signal. The hg-vap-2 cDNA contained an open reading frame encoding 212 amino acids with the first 19 amino acids being a putative secretion signal. Genes of hg-vap-1 and hg-vap-2 contained four introns, which ranged in size from 44 to 574 bp, and five exons ranging in size from 43 to 279 bp. In situ hybridisation analyses showed that mRNAs of both vap genes accumulated specifically in the subventral gland cells of H. glycines during parasitism. The gland cell-specific expression and presence of predicted secretion signal peptides in both VAPs suggest that these proteins are secreted from the nematode and may play a role in the infection of host plants by this parasite. PMID- 11730790 TI - The effect of challenge and trickle Trichuris muris infections on the polarisation of the immune response. AB - In the field, determination of mechanisms of immunity to geohelminths are problematic due to the variation in infection exposure, host genetics, nutrition and co-infection. This study uses a well defined laboratory model, Trichuris muris in the mouse to study immune responses to challenge and trickle infections. The rationale is thus to study parasite acquisition under more natural antigen dose exposure. Antigen dose has previously been shown in this system to affect the outcome of infection with low antigen doses favouring type 1 responses (and susceptibility) and high antigen doses favouring type 2 responses (and resistance). A high level challenge infection could be established in a normally resistant host but only following priming of the immune response by a low level infection. Once type 2 responses were initiated it was impossible to switch an ongoing type 2 response even using IL-12 which is a potent stimulus of type 1 responses. Trickle infections resulted in no clear polarisation of the immune response. It was possible to build up the level of infection to a threshold level beyond which type 2 responses and expulsion were initiated. This threshold level was dependent upon host genetic background. Our results reveal a complex spectrum of responses and demonstrate that resistance and type 2 responses can be built up with increasing parasite exposure. The data provide compelling evidence to support a role for acquisition of acquired immunity to gastro-intestinal nematodes under complex infection patterns such as those found in the field. PMID- 11730791 TI - Reduction of transmission stages concomitant with increased host immune responses to hypervirulent Sarcocystis singaporensis, and natural selection for intermediate virulence. AB - Parasite virulence (pathogenicity depending on inoculum size) and host immune reactions were examined for the apicomplexan protozoan Sarcocystis singaporensis. This parasite is endemic in southeastern Asia and multiplies as a proliferation (merozoite) and transmission stage (bradyzoite) in rats. Virulence in wild brown rats of parasites freshly isolated in the wild (wild-type) was surprisingly constant within the endemic area and showed an intermediate level. In contrast, serially passaged parasites either became avirulent or virulence increased markedly (hypervirulence). Production of transmission stages was maximal for the wild-type whereas numbers were significantly reduced for hypervirulent and avirulent (shown in a previous study) parasites. Analyses of B and T cell immunity revealed that immune responses of WKY rats to the transmission stage were significantly higher for hypervirulent than for wild-type parasites. These results suggest that it is the immune system of the host that is not only responsible for reduction of transmission stages in individual rats, but also could act as a selective force that maintains intermediate virulence at the population level because reduction of muscle stages challenges transmission of S. singaporensis to the definitive host. Collectively, the presented data support evolutionary theory, which predicts intermediate rates of parasite growth in nature and an 'arms race' between host immunity and parasite proliferation. PMID- 11730792 TI - Eosinophils in the cerebrospinal fluid of mice infected with Angiostrongylus cantonensis are resistant to apoptosis. AB - Interleukin-5 (IL-5) transgenic mice were used to assess the immunological features of CSF eosinophils from mice infected with Angiostrongylus cantonensis. CSF eosinophils were hypodense by day 14 post infection (p.i.). CSF eosinophils survived longer in vitro than peritoneal eosinophils collected from cadmium sulphate (CdSO(4)) -treated normal IL-5 transgenic mice. Apoptosis was measured by Annexin V binding and the presence of a distinct laddering pattern of DNA fragmentation on agarose electrophoresis. Regardless of the presence or absence of Actinomycin D, CSF eosinophils collected from IL-5 transgenic mice from days 15-36 p.i. exhibited less apoptosis than peritoneal eosinophils collected from uninfected IL-5 transgenic mice. CSF eosinophils collected from A. cantonensis infected C57BL/6 mice at days 15-34 p.i. showed elongation of survival time and less apoptosis during in vitro cultivation. Reduced apoptosis was noted only in CSF eosinophils, but not in peritoneal eosinophils recovered from the same infected IL-5 transgenic mice. CPP32/Caspase 3 activity of cultured peritoneal eosinophils from both infected and uninfected IL-5 transgenic mice was higher than that of cultured CSF eosinophils. Stimulation with A23187 readily induced apoptosis of peritoneal eosinophils, but not CSF eosinophils or peritoneal eosinophils cultured with mouse recombinant IL-5. The latter cells were morphologically identical to hypodense eosinophils. RT-PCR analysis indicated that bcl-2 and bcl-x(L) mRNA expression was higher in CSF eosinophils compared with peritoneal eosinophils and this expression in the latter cells was upregulated after culture with mouse recombinant IL-5. These results suggest that CSF eosinophils, shifting to hypodense status through an accumulation from peripheral blood, are resistant to apoptosis. These changes may explain the long lasting, helminthotoxic and neurotoxic actions of CSF eosinophils in A. cantonensis infection. PMID- 11730793 TI - Identification of a human immunodominant B-cell epitope within the GRA1 antigen of Toxoplasma gondii by phage display of cDNA libraries. AB - Excreted secreted antigens of the protozoan parasite Toxoplasma gondii play a key role in stimulating the host immune system during acute and chronic infection. With the aim of identifying the immunodominant epitopes of T. gondii antigens involved in the human B-cell response against the parasite, we employed a novel immunological approach. A library of cDNA fragments from T. gondii tachyzoites was displayed as fusion proteins to the amino-terminus of lambda bacteriophage capsid protein D. The lambda D-tachyzoite library was then affinity-selected by using a panel of sera of pregnant women, all infected with the parasite. Some of the clones identified through this procedure matched the sequence of the dense granule GRA1 protein (p24), allowing us to identify its antigenic regions. In particular, the analysis of human antibody response against the recombinant GRA1 antigen fragments revealed the existence of an immunodominant epitope (epi-24 peptide). PMID- 11730794 TI - Allelic variation in the cg2 gene does not correlate with chloroquine resistance among Indian Plasmodium falciparum isolates. AB - The cg2 gene of Plasmodium falciparum has been proposed to be associated with chloroquine resistance. Here we describe PCR amplification and sequencing of all the four repeat regions (kappa (kappa), gamma (gamma), psi (psi) and omega (omega)) of this gene, from Indian isolates. There were variant forms for each of these repeat regions (two for kappa and gamma, and three for psi and omega) among the 123 Indian isolates of P. falciparum. Among these isolates certain forms of psi and omega repeats were uniquely present while some of the reported forms of the kappa and omega repeats were absent. The pattern of combination of all four repeat regions of cg2 gene (genotype) was analysed from 52 isolates. A total of 11 different genotypes were observed among these cases, of which 10 were unique to Indian isolates. Certain genotypes were more common than others. The nucleotide sequencing of all the four repeat regions revealed that Indian isolates have some unique repeating units within the gamma and omega domains. Altogether, the PCR and sequencing results showed that there was an unrelatedness between cg2 repeats and chloroquine resistance. PMID- 11730795 TI - Characterisation of the gene encoding a protective antigen from Babesia microti identified it as eta subunit of chaperonin containing T-complex protein 1. AB - Passive immunisations with a monoclonal antibody termed 1-5H showed a partial but significant inhibition of parasitaemia against Babesia microti challenge infection. By immunoscreening with 1-5H, a clone (termed p58 gene) was obtained from a cDNA expression library of B. microti and the complete nucleotide sequence was determined. A protein homology search showed significant amino acid identities to the eta subunit of the chaperonin containing T-complex protein 1 (CCT) of human (59%), mouse (58%) and Plasmodium falciparum (62%). Genomic analyses indicated that the p58 gene is present as a single copy gene and contains a total of approximately 400-bp introns in the genome of B. microti. The mAb 1-5H recognised a 58-kDa protein of B. microti and was found to cross-react with a 60-kDa protein of Babesia rodhaini. These results suggest the possibility that the p58 protein is the CCT eta subunit of B. microti and functions as a chaperonin. PMID- 11730796 TI - Identification of two putative ATP-cassette genes in Encephalitozoon intestinalis. AB - Currently existing chemotherapeutic compounds are limited and few are effective for treating microsporidiosis. It is possible that resistance of Encephalitozoon to some drugs occurs by efflux mechanisms similar to those previously described for mammalian tumour cells, bacteria or protozoal parasites such as Plasmodium, Leishmania and Entamoeba histolytica. The data in the present study suggest that Encephalitozoon intestinalis contains at least one multidrug resistance gene. We report here two complete sequences EiABC1 and EiABC2, encoding different ATP binding cassette genes from E. intestinalis, including a P-gp. PMID- 11730797 TI - Licking behaviour and environmental contamination arising from pour-on ivermectin for cattle. AB - Pour-on formulations of endectocides are extensively used to treat and control systemic parasitic diseases in cattle, worldwide. The purpose of the present study was to investigate the influence of the natural licking behaviour of cattle on the plasma and faecal disposition of topically administered ivermectin. Twelve Holstein cattle were given one single intravenous (i.v.) (200 microg/kg) and topical (500 microg/kg) administration of ivermectin at a 5-month interval. For the pour-on administration, the animals were allocated into two groups (n=6): one control group (lickers) and one group where licking was prevented (non-lickers). Ivermectin plasma (total) clearance (270+/-57.4 ml/kg/day) was very homogeneous among the 12 cattle. In contrast, major differences between lickers and non lickers were observed following pour-on administration. Prevention of licking resulted in an extended terminal plasma half-life (363+/-16.2 vs. 154+/-7.4 h in lickers) and in a lower and less variable systemic availability of ivermectin (19+/-4.9 vs. 33+/-18.5% in lickers). More importantly, nearly 70% of the pour-on dose was recovered as parent drug in the faeces of lickers vs. only 6.6% in non lickers. Altogether, these results are consistent with an oral rather than percutaneous absorption of topical ivermectin in control animals, the non systemically available fraction of ingested ivermectin providing a major contribution (80%) to the drug faecal output. The consequences of licking on the disposition of pour-on ivermectin are discussed in terms of environment, given the known ecotoxicity of this drug, and of cross-contamination. Animals licking themselves and each other could result in unexpected residues in edible tissues of untreated animals and in possible subtherapeutic drug concentrations, a factor in drug resistance. According to the Precautionary Principle, these considerations elicit concern over the use of topical drug formulations in cattle. PMID- 11730798 TI - Differentiation among three species of bovine Thelazia (Nematoda: Thelaziidae) by polymerase chain reaction-restriction fragment length polymorphism of the first internal transcribed spacer ITS-1 (rDNA). AB - Thelazia gulosa, Thelazia rhodesi and Thelazia skrjabini are nematodes transmitted by some species of Musca (Diptera: Muscidae) which cause ocular infestations in bovines. Differences in the rDNA of these species were determined by a PCR using different sets of relatively conserved oligonucleotide primers. PCR on the first internal transcribed spacer (ITS-1) revealed differences in size in Thelazia species (437 bp for T. gulosa, 370 bp for T. rhodesi and 506 bp for T. skrjabini) while the DNA control of Musca spp. was not amplified. The ITS-1 amplicons of the three species were sequenced and then analysed. The GC contents ranged from 26 to 36% and the level of differences in the nucleotide sequences of ITS-1 was lower between T. skrjabini and T. gulosa (39%) than the latter and T. rhodesi (49-56%). Restriction fragment length polymorphism (RFLP) of ITS-1 amplicons was also carried out and the restriction profiles compared. Clear genetic differences among the three Thelazia examined were demonstrated by using the enzymes HpaII, CpoI and SspI. This PCR-RFLP for the delineation of T. gulosa, T. rhodesi and T. skrjabini offers prospects as a molecular epidemiological tool to study parasite transmission patterns and prevalence. PMID- 11730799 TI - High levels of congenital transmission of Toxoplasma gondii in a commercial sheep flock. AB - Our current understanding of congenital transmission of Toxoplasma gondii from ewe to lamb dictates that infection frequently results in abortion and the death of the developing foetus, that the birth of live infected lambs occurs rarely and that the cat is the predominant source of infection in ewes. Using direct polymerase chain reaction detection of T. gondii, we report high levels of congenital transmission occurring in a commercially managed sheep flock. We sampled foetal-derived placental tissue and tissues from aborted lambs and showed that congenital transmission was detected in these tissues from 61% of all pregnancies. Where pregnancies resulted in the death of one or more lambs, T. gondii was detected in the lamb tissue for all but one of 18 (94%) pregnancies. Of the successful pregnancies resulting in the birth of live lambs we were able to detect T. gondii in foetal-derived placental tissue from 37 of 70 (42%) pregnancies. These results show that congenital transmission is occurring in a high percentage of lambings including normal healthy lambings, at this farm, suggesting that this route of transmission from generation to generation may be much more significant than that reported previously. These results may have implications for sheep husbandry and future epidemiological studies of T. gondii. PMID- 11730800 TI - Major surface protein 1a effects tick infection and transmission of Anaplasma marginale. AB - Anaplasma marginale, an ehrlichial pathogen of cattle and wild ruminants, is transmitted biologically by ticks. A developmental cycle of A. marginale occurs in a tick that begins in gut cells followed by infection of salivary glands, which are the site of transmission to cattle. Geographic isolates of A. marginale vary in their ability to be transmitted by ticks. In these experiments we studied transmission of two recent field isolates of A. marginale, an Oklahoma isolate from Wetumka, OK, and a Florida isolate from Okeechobee, FL, by two populations of Dermacentor variabilis males obtained from the same regions. The Florida and Oklahoma tick populations transmitted the Oklahoma isolate, while both tick populations failed to transmit the Florida isolate. Gut and salivary gland infections of A. marginale, as determined by quantitative PCR and microscopy, were detected in ticks exposed to the Oklahoma isolate, while these tissues were not infected in ticks exposed to the Florida isolate. An adhesion-recovery assay was used to study adhesion of the A. marginale major surface protein (MSP) 1a to gut cells from both tick populations and cultured tick cells. We demonstrated that recombinant Escherichia coli expressing Oklahoma MSP1a adhered to cultured and native D. variabilis gut cells, while recombinant E. coli expressing the Florida MSP1a were not adherent to either tick cell population. The MSP1a of the Florida isolate of A. marginale, therefore, was unable to mediate attachment to tick gut cells, thus inhibiting salivary gland infection and transmission to cattle. This is the first report of MSP1a being responsible for effecting infection and transmission of A. marginale by Dermacentor spp. ticks. The mechanism of tick infection and transmission of A. marginale is important in formulating control strategies and development of improved vaccines for anaplasmosis. PMID- 11730804 TI - Endogenous heparin activity deficiency: the 'missing link' in atherogenesis? AB - This paper reviews published studies since 1995 dealing with many atherogenic mechanisms where exogenous heparin was beneficial. In these areas endogenous heparin deficiency is likely to be harmful. Mechanisms included inflammatory factors, lower endogenous plasma heparin levels, lipoprotein lipase, chemokines, APOE e4, lipoprotein(a), among others. Demonstrated reduction of heparan sulfate proteoglycans (HSPG) and of endogenous plasma heparin was reviewed. PMID- 11730805 TI - Loss of heterozygosity of the NOS3 dinucleotide repeat marker in atherosclerotic plaques of human carotid arteries. AB - We have investigated 28 atherosclerotic plaques of human carotid arteries with a panel of 39 microsatellite markers for the presence of LOH. The objective of this research was to verify if LOH, described in association with tumorigenic process, could be involved also in benign fibroproliferative disease. Seventy percent of samples demonstrated allelic imbalance: 50% of cases showed LOH at a minimum of one locus, 3.5% at a minimum of two loci and 14.3% at three or more loci. The percentages of LOH ranged between 3.8 and 14.3% and the highest involved polymorphic marker is the NOS3 internal dinucleotide repeat. Our results indicate that, like tumorigenesis, the atherogenic process could also involve LOH mechanism. Furthermore, the finding regarding the NOS3 internal polymorphism suggests a possible role of the gene as cofactor in formation of the atheromas. PMID- 11730806 TI - Serotonin potentiates angiotensin II--induced vascular smooth muscle cell proliferation. AB - Vascular smooth muscle cell (VSMC) proliferation is a key feature in the development of atherosclerosis and restenosis after angioplasty, which can occur in response to many different humoral and mechanical stimuli. We investigated the growth promoting activities of two potent vasoactive substances, angiotensin II (Ang II) and serotonin (5-HT), on cultured rabbit VSMCs. Growth-arrested VSMCs were incubated with serum-free medium containing different concentrations of Ang II in the presence or absence of 5-HT. [3H]thymidine incorporation into VSMC DNA was measured as an index of cell proliferation. Ang II and 5-HT stimulated DNA synthesis in a dose-dependent manner with a maximal effect at 1.75 microM for Ang II (202%) and 50 microM for 5-HT (205%). When added together, low concentrations of Ang II (1 microM) and 5-HT (5 microM) synergistically induced DNA synthesis (363%). Candesartan (1 microM), an AT(1) receptor antagonist, but not PD 123319 (1 microM), an AT(2) receptor antagonist, inhibited the mitogenic effect on Ang II and its interaction with 5-HT. Sarpogrelate (10 microM), a 5-HT(2A) receptor antagonist, and pertussis toxin (10 ng/ml) inhibited the mitogenic effect of 5-HT and its interaction with Ang II. The protein kinase C inhibitor Ro 31-8220 (0.1 microM), the Raf-1 inhibitor radicicol (10 microM), and the MAPK kinase inhibitor PD 098059 (10 microM) abolished mitogenic effects of Ang II and 5-HT, and also their synergistic interaction. The JAK2 inhibitor AG 490 (10 microM) had only a minimal inhibitory effect of Ang II-induced DNA synthesis but significantly inhibited the interaction of Ang II with 5-HT. The synergistic effect on Ang II (1 microM) with 5-HT (5 microM) on DNA synthesis was completely reversed by the combined use of both candesartan (1 microM) and sarpogrelate (10 microM). Our results suggest that Ang II and 5-HT exert a synergistic interaction on VSMC proliferation via AT(1) and 5-HT(2A) receptors. The activation of MAPK and JAK/STAT pathways may explain the synergistic interaction between Ang II and 5 HT. PMID- 11730807 TI - Age dependent aneuploidy and telomere length of the human vascular endothelium. AB - RATIONALE: Aneuploidy and telomere length are two major parameters that have been associated with cellular senescence in vitro. In order to explore the role of aneuploidy and telomere length in aging of the human vasculature, we studied these two parameters in direct preparations of endothelial cells of the human abdominal aorta. METHODS: Using fluorescent in situ hybridization on 'touch prep' slides, we evaluated aneuploidy of two autosomes (chromosomes 6 and 16) and sex chromosomes in non cultured endothelial cells of the abdominal aorta as a function of the donor's age. RESULTS: We found that the frequency of aneuploidy of vascular endothelial cells significantly increased with age. This was expressed by age-dependent tetrasomy (r(s)=0.56, P=0.006 for chromosome 6; and r(s)=0.54, P=0.008 for chromosome 16), and age dependent loss of the Y chromosome (r(s)=0.85, P=0.0003). In addition, we found that telomere length was inversely correlated with age (r=-0.38, P=0.008). DATA INTERPRETATION: These findings suggest that indicators of cellular senescence, earlier observed in vitro, are also expressed in the human vascular endothelium in vivo. Aneuploidy and telomere attrition might thus play a role in the aging of the human vasculature. PMID- 11730808 TI - Molecular sieving and mass spectroscopy reveal enhanced collagen degradation in rabbit atheroma. AB - BACKGROUND: Collagen degradation is the major mechanism of atherosclerotic plaque destabilization. It is unknown whether collagen breakdown is involved into formation of early atherosclerotic lesions. METHODS: Current paper describes a novel collagen degradation assay based on a combination of molecular sieving and mass spectroscopy. The first step of the assay consists of the extraction of total collagen from tissue. This extract includes both intact collagen and its breakdown products. Molecular sieving is used to isolate low molecular weight collagen fragments. Since the low molecular weight fraction of the extract may contain some non-collagenous molecular species, the collagen-specific amino acid hydroxyproline is quantified using mass spectroscopy. RESULTS: This assay was validated in various experimental systems with known/predictable level of collagen breakdown in vitro, ex vivo and in vivo. When applied to cholesterol-fed rabbit aorta, it revealed enhanced collagen degradation in rabbit atheromas compared to unaffected aortic regions. CONCLUSION: A novel assay has been developed to demonstrate enhanced collagen degradation in rabbit atherosclerotic plaques. Accurate quantification of collagen breakdown products should provide a new relevant end point in the analysis of plaque development and stability. PMID- 11730809 TI - Urokinase plasminogen activator augments cell proliferation and neointima formation in injured arteries via proteolytic mechanisms. AB - Urokinase plasminogen activator (uPA) has been implicated in the healing responses of injured arteries, but the importance of its various properties that influence smooth muscle cell (SMC) proliferation and migration in vivo is unclear. We used three recombinant (r-) forms of uPA, which differ markedly in their proteolytic activities and abilities to bind to the uPA receptor (uPAR), to determine, which property most influences the healing responses of balloon catheter injured rat carotid arteries. After injury, uPA and uPAR expression increased markedly throughout the period when medial SMCs were rapidly proliferating and migrating to form the neointima. Perivascular application of uPA neutralizing antibodies immediately after injury attenuated the healing response, significantly reducing neointima size and neointimal SMC numbers. Perivascular application of r-uPAwt (wild type uPA) or r-uPA/GDF (r-uPA with multiple mutations in its growth factor-like domain) doubled the size of the neointima. Four days after injury these two uPAs nearly doubled neointimal and medial SMC numbers in the vessels, and induced greater reductions in lumen size than injury alone. Proteolytically inactive r-uPA/H/Q (containing glutamine rather than histidine-204 in its catalytic site) did not affect neointima or lumen size. Also, in contrast to the actions of proteolytically active uPAs, tissue plasminogen activator (tPA) did not affect the rate of neointima development. We conclude that uPA is an important factor regulating the healing responses of balloon catheter injured arteries, and its proteolytic property, which cannot be mimicked by tPA, greatly influences SMC proliferation and early neointima formation. PMID- 11730810 TI - Heparin-stimulated expression of extracellular-superoxide dismutase in human fibroblasts. AB - Extracellular-superoxide dismutase (EC-SOD) is the major SOD isozyme in the arterial wall and may be important for antioxidation capability of the vascular wall and normal vascular function. EC-SOD is expressed in various cell types in the vascular wall such as fibroblasts, smooth muscle cells and macrophages, and the synthesis of EC-SOD by human fibroblasts is known to be highly responsive to various inflammatory cytokines, although there is no response to oxidative stress. Heparin is a highly sulfated glycosaminoglycan with many functions such as antithrombotic, antilipemic and antiatherosclerotic effects. Another less well known function of heparin is regulation of protein synthesis. In this study, we measured the induction of EC-SOD after treatment with heparin to understand the role of heparin in the antiatherosclerotic response of fibroblasts. Heparin induced EC-SOD expression at both the mRNA and protein levels. Heparin showed the greatest stimulatory effect and heparan sulfate showed moderate effects. The effect of chondroitin sulfate A was not clear. In contrast, desulfated heparin and chondroitin sulfate C did not increase EC-SOD expression. The stimulatory effect seemed to increase roughly with the degree of glycosaminoglycan sulfation. The enhanced expression of EC-SOD by heparin must contribute to the antiatherosclerotic effect of heparin. PMID- 11730811 TI - Apolipoprotein AI and HDL(3) inhibit spreading of primary human monocytes through a mechanism that involves cholesterol depletion and regulation of CDC42. AB - The objective of the current study was to characterize the influence of high density lipoproteins (HDL) on processes related to the vascular recruitment of human monocytes, which may contribute to the anti-atherogenic properties of these lipoproteins. We show that HDL(3) and apo AI inhibit the following processes in primary human monocytes: (1) M-CSF induced cell spreading; (2) M-CSF stimulated expression of surface molecules involved in adhesion, migration, and scavenging; (3) fMLP induced chemotaxis. These processes are obviously modulated by the regulation of cellular cholesterol pools as indicated by the following findings. In Tangier monocytes with defective apo AI induced cholesterol efflux, apo AI had no influence on the spreading response. In control cells, stimulation of cholesterol efflux by p-cyclodextrin mimicked the effect of apo AI and HDL(3) on spreading and chemotaxis, whereas cholesterol loading with enzymatically modified LDL (E-LDL) showed the opposite effect. Finally, a similar inverse regulation by E-LDL and apo AI/HDL(3) was also observed in regard to the surface expression of beta(1)- and beta(2)-integrins as well as the hemoglobin/haptoglobin scavenger receptor CD163 and the Fcgamma-IIIaR CD16. CDC42 was identified as a potential downstream target linking changes in cellular cholesterol content to monocyte spreading and chemotaxis. Thus, CDC42 antisense markedly reduced spreading and, in parallel with their influence on monocyte spreading, HDL(3), apo AI and p cyclodextrin down-regulated CDC42 expression while E-LDL had the inverse effect. The apo AI induced decrease of CDC42 protein expression was paralleled by the reduction of active GTP-bound CDC42. In summary, we provide evidence that HDL(3) and apo AI are able to inhibit processes in primary human monocytes, which are related to the recruitment of monocytes into the vessel wall and probably involve regulation of cellular cholesterol pools and CDC42 function. PMID- 11730812 TI - Vascular endothelial growth factor synthesis in vascular smooth muscle cells is enhanced by 7-ketocholesterol and lysophosphatidylcholine independently of their effect on nitric oxide generation. AB - Nitric oxide (NO) generated by inducible NO synthase (iNOS) enhances vascular endothelial growth factor (VEGF) synthesis in vascular smooth muscle cells (VSMC) and both NO and modified low density lipoprotein (LDL) augment VEGF production in macrophages. Oxidized LDL (oxLDL) are known inhibitors of NO generation in the cells of vascular wall. As the relationship between VEGF, iNOS and oxLDL has not been well elucidated, we studied the effect of two main components of oxLDL, 7 ketocholesterol (7-Kchol) and lysophosphatidylcholine (LPC), on VEGF and NO synthesis in rat VSMC and on VEGF synthesis in human VSMC. Both LPC and 7-Kchol significantly augmented VEGF production in rat and human VSMC. Increase in VEGF generation was related to the activation of VEGF promoter by both 7-Kchol and LPC and enhancement of VEGF mRNA transcription. In rat, VSMC IL-1beta-induced NO generation and enhanced VEGF synthesis. 7-Kchol decreased rat iNOS promoter activity, iNOS expression and NO generation, but it did not impair IL-1beta induced VEGF synthesis. LPC did not significantly influence IL-1beta-induced NO production in rat VSMC and VEGF synthesis was significantly enhanced by combined treatment with IL-1beta and LPC in comparison to the effect of either compound alone. The results indicate that VEGF and NO synthesis in VSMC can be modulated by oxLDL. Those interactions might have an effect on the plaque growth and might be of relevance for the physiology of vascular wall cells. PMID- 11730813 TI - Influence of non-enzymatically glycated collagen on monocyte-macrophage differentiation. AB - Blood monocytes (mo) on transendothelial migration interact with extracellular matrix components (ECM) and differentiate into macrophages (m(phi)), which play an important role in both physiological, and pathological conditions, particularly, atherosclerosis. In order to study whether modification of ECM such as non-enzymatic glycation occurring in diabetes influences mo-m(phi) differentiation, an in vitro system using isolated human peripheral blood mononuclear cells (PBMC) maintained on non-enzymatically glycated COL I (NEG COL I) was used. M(phi) specific functions such as receptor mediated endocytosis of modified proteins, production of m(phi) specific 92 and 72 kDa matrix metalloproteinases (MMPs), expression of surface antigen and loss of myeloperoxidase (MPO) activity were assessed. Endocytosis of 125[I] acetyl BSA was significantly higher in cells maintained on NEG COL I than those on COL I. Kinetic analysis revealed that the rate of uptake of modified BSA and production of MMPs by cells maintained on NEG COL I were higher than those on COL I suggesting a faster rate of differentiation of cells maintained on modified substrata. FACS analysis of the expression of surface antigen showed that the rate of down-regulation of monocyte specific CD14 and the rate of up-regulation of m(phi) specific CD71 were high in cells maintained on NEG COL I. These results suggest that the interaction of monocyte with non-enzymatically glycated matrix protein in the vessel wall may result in faster rate of induction of mo-m(phi) differentiation leading to foam cell formation, a critical early event in the initiation and development of atherosclerosis. PMID- 11730814 TI - GM-CSF: a strong arteriogenic factor acting by amplification of monocyte function. AB - We investigated the role of the colony stimulating factor for monocytes (GM-CSF) to test the hypothesis whether prolongation of the monocyte's life cycle will support arteriogenesis (rapid growth of preexisting collateral arteries). This appeared logical in view of our discovery that circulating monocytes play an important part in the positive remodeling of small preexisting arterioles into arteries to compensate for arterial occlusions (arteriogenesis) and especially following our findings that MCP-1 markedly increases the speed of arteriogenesis. The continuous infusion of GM-CSF for 7 days into the proximal stump of the acutely occluded femoral artery of rabbits by osmotic minipump produced indeed a marked arteriogenic response as demonstrated by an increase (2-fold) in number and size of collateral arteries on postmortem angiograms and by the increase of maximal blood flow during vasodilation measured in vivo by blood pump perfusion of the hindquarter (5-fold). When GM-CSF and MCP-1 were simultaneously infused the effects on arteriogenesis were additive on angiograms as well as on conductance. GM-CSF was also able to widen the time window of MCP-1 activity: MCP 1 treatment alone was ineffective when given after the third week following occlusion. When administered together with GM-CSF about 80% of normal maximal conductance of the artery that was replaced by collaterals were achieved, a result that was not reached before by any other experimental treatment. Experiments with cells isolated from treated animals showed that monocyte apoptosis was markedly reduced. In addition we hypothesize that GM-CSF may aid in releasing pluripotent monocyte (stem-) cells from the bone marrow into the circulation. In contrast to MCP-1, GM-CSF showed no activity on monocyte transmigration through- and also no influence on monocyte adhesion to cultured endothelial cells. In conclusion we have discovered a new function of the hemopoietic stem cell factor GM-CSF, which is also a powerful arteriogenic peptide that acts via prolongation of the life cycle of monocytes/macrophages. PMID- 11730815 TI - Hormone replacement therapy and endothelial function. Results of a randomized controlled trial in healthy postmenopausal women. AB - OBJECTIVE: To compare the effects of 3 months treatment with tibolone (a single entity synthetic steroid hormone with estrogenic, progestanic and androgenic activities), or continuous combined conjugated equine estrogens (CEE) plus medroxyprogesterone acetate (MPA), with placebo, on endothelial function. DESIGN: A single center, randomized, double-blind, placebo-controlled study. SETTING: Research center as part of the University Medical Center Utrecht. SUBJECTS: One hundred and five healthy postmenopausal women, sampled from the general population. INTERVENTIONS: Three months treatment with tibolone or CEE+MPA or placebo. MAIN OUTCOME MEASURE: At baseline and after 3 months, endothelial function was assessed non-invasively by measuring percent lumen diameter change in the brachial artery after reactive hyperemia and sublingual nitroglycerine spray. RESULTS: Results are presented as mean differences between treatment groups of endothelium dependent flow mediated dilatation (fmd) and endothelium independent nitroglycerine induced dilatation with 95% confidence intervals (95% CI). After treatment, there was a significant difference in mean fmd between the CEE+MPA group and the placebo group of 2.5% (95% CI: 0.3-4.6) while the tibolone group and the placebo group did not differ significantly (0.6%; 95% CI: 1.6-2.8). Nitroglycerine induced dilatation did not differ significantly between the groups. CONCLUSIONS: Hormone replacement therapy with CEE+MPA for 3 months increases endothelium dependent fmd of the brachial artery in healthy postmenopausal women. Tibolone did not alter fmd. The clinical significance of this improvement in fmd for cardiovascular disease risk needs to be established. PMID- 11730816 TI - Influence of lipoprotein lipase serine 447 stop polymorphism on tracking of triglycerides and HDL cholesterol from childhood to adulthood and familial risk of coronary artery disease: the Bogalusa heart study. AB - The effects of the lipoprotein lipase (LPL) Serine 447 Stop (S447X) polymorphism on high-density lipoprotein cholesterol (HDLC) and triglycerides (TG) have been demonstrated. However, little is known about its effect on the tracking of HDLC and TG over time and familial risk of coronary artery disease (CAD). This aspect was examined in black and white individuals (n=829) aged 5-18 year at baseline, followed on average 18.8 yr. The frequency of the X447 allele was lower in Blacks than Whites (0.043 vs. 0.087, P=0.002). Carriers vs. noncarriers of the X447 allele had lower TG (99.3 vs 122.1 mg/dl, P<0.01) and higher HDLC (51.1 vs. 49.7 mg/dl, P<0.05) in adulthood, but not in childhood. The trends in genotype specific means of childhood and adulthood levels of HDLC and TG in sex or race subgroups were similar to those in the total sample. With respect to tracking over time, of those in the bottom quartile of HDLC in childhood, 46.1% of the noncarriers vs. 23.1% of the carriers remained in this lowest quartile into adulthood (P=0.03); corresponding values for the top quartile of HDLC were 37.5% for the noncarriers vs. 57.1% for the carriers (P=0.03). Although TG tended to track better among the carriers in the bottom quartile and among the noncarriers in the top quartile, this trend was not significant. Carriers showed lower prevalence of parental history of CAD than noncarriers (6.9% vs. 14.1%, P=0.02) independently of lipoprotein variables, adiposity, blood pressure, age, sex and race. Thus, the X447 allele of the LPL gene is associated with an increase in HDLC and a decrease in TG in adults, tracking of HDLC since childhood, and a lower family history of CAD. PMID- 11730817 TI - No evidence of accelerated atherosclerosis in a 66-yr-old chylomicronemia patient homozygous for the nonsense mutation (Tyr61-->stop) in the lipoprotein lipase gene. AB - Whether chylomicronemia is atherogenic or not has yet to be determined in humans. We investigated a 66-yr-old female with severe chylomicronemia resulting from a lipoprotein lipase (LPL) deficiency. The patient's plasma triglyceride level was approximately 2000 mg/dl. Both LPL activity and the mass of postheparin plasma in this patient were virtually absent. A nonsense mutation in exon 3 (Tyr61-->Stop) was identified in the patient's LPL gene, and a restriction fragment length polymorphism analysis established that the patient was homozygous for this mutation. The patient was neither a diabetic nor a smoker. Clinically, the patient had never experienced pancreatitis or angia pectoris. An examination of her carotid, femoral and coronary arteries by ultrasonogram and electrocardiogram after exercise-tolerance testing showed no accelerated atherosclerosis. This case suggests that atherosclerosis may not occur despite massive hyperlipidemia, when LPL bridging was not present due to the absence of LPL secretion and circulating mass. PMID- 11730818 TI - Studies on the plasma lipid profiles, and LCAT and CETP activities according to hyperlipoproteinemia phenotypes (HLP). AB - Hyperlipoproteinemia phenotypes (HLP), one of genetic disorders with an estimated prevalence of 0.5-2% in the general population, is responsible for 10% of premature CHD. After first screening with the high cholesterol (>6.47 mM/l) and triglyceride (TG) (>2.6 mM/l) levels without medication, subjects were typed for HLP classification. Differential metabolic effects of HLP types on plasma lipid profiles and the reverse cholesterol transport system (RCT) were studied in 196 HLP types (91.2%) and 19 non-HLP (8.8%). A total of 45% of subjects had primary HLP and the others had NIDDM (10.7%), hypertension (9.3%) and other chronic diseases. Type IV HLP (58.6%) was most predominant and Types IIa, IIb, III and V comprised 16.7, 12.1, 2.3 and 1.4% of the HLP. Type I was not found. Plasma lipids excluding apo A-I and Lp(a) were significantly different among HLP compared to non-HLP (P<0.001). Since Type V and III impact the clearance of TG riched lipoproteins, TG and VLDL-C levels were higher in V and III. TG and LDL-C were higher in Type II than those in the others because of defect of LDL receptors. LCAT activity, lower in Type III and Type IV and highest in Type V, was highly associated with plasma free cholesterol levels and the ratio of apoB/apoA and LDL/HDL. CETP activity was highest in Type V due to high VLDL-C and TG and low HDL-C. The ratio of LCAT/CETP was not different among HLP types but was significantly lower in HLP than in non-HLP. CETP increased 2-3 times as well as LCAT decreased among HLP patients compared to non-HLP. We conclude non-HLP subjects with high cholesterol and TG levels do not always mean high risk of CHD and the intervention effects of HLP types would lead to impose the risk of CHD by the impact of RCT. PMID- 11730819 TI - Distribution and correlates of lipoproteins and their subclasses in black and white young adults. The Bogalusa Heart Study. AB - Lipoprotein subclasses vary in CAD risk potential, but their distribution and correlates are not well documented in black and white young adults. A subsample of 449 (32%) young adults (67% white, 58% female) aged 20-37 years examined in the Bogalusa Heart Study had lipoprotein subclasses measured in terms of cholesterol by vertical spin density-gradient ultracentrifugation. LDL subclass pattern was characterized as either predominantly LDL(1) (large, buoyant), LDL(2) (intermediate) or LDL(3) (small, dense). Whites had significantly higher levels of VLDL, VLDL(3), and LDL and lower levels of HDL(2) and HDL(3) than blacks. White females had significantly higher levels of HDL(2) than white males. Visceral fatness, measured as waist circumference, and race were the major contributors to the explained variance (6-22%) of these lipoproteins, with adverse trends seen among whites and persons with large waist circumferences. Sex (males>females), waist circumference (positive), HDL(2) (negative), and HDL(3) (positive) were the predictor variables for the likelihood of having the LDL(3) pattern. When glucose and insulin were included in the multivariate analysis, insulin (positive), sex (males>females), HDL(2) (negative) and HDL(3) (positive) became significant predictors of LDL(3) pattern. Positive parental history of CAD was associated with LDL (P=0.009) in white males, and HDL(2) (P=0.008) and LDL(3) subclass pattern (P=0.038) in white females; whereas none in blacks. The observed correlates of lipoprotein subclasses and patterns need to be considered in estimating CAD risk in young adults. PMID- 11730820 TI - Race differences in the response of postheparin plasma lipoprotein lipase and hepatic lipase activities to endurance exercise training in men: results from the HERITAGE Family Study. AB - Endurance exercise training is known to produce favorable changes in the metabolic profile including reduced plasma triglyceride (TG) and increased high density lipoprotein (HDL) cholesterol concentrations. These metabolic improvements are likely to contribute to the reduced coronary heart disease (CHD) risk often observed in physically active individuals. However, the physiological mechanisms responsible for such improvements in TG and HDL cholesterol concentrations with endurance exercise are not fully understood. The effect of a 20-week endurance exercise training program on plasma lipoproteins as well as on post-heparin plasma lipoprotein lipase (PH-LPL) and hepatic lipase (PH-HL) activities were therefore examined in a sample of 200 White and 69 Black men who were part of the HERITAGE Family Study. As expected, there were decreases in adiposity and in abdominal fat accumulation following training in both White and Black men. We also found that exercise training was associated with decreases in plasma cholesterol, TG and apolipoprotein B levels, as well as with an increase in HDL cholesterol concentrations in White men. In contrast, Black men showed an increase only in HDL(2) cholesterol over the 20-week period. Higher PH-LPL and lower PH-HL activities were noted in both ethnic groups at follow-up. Whereas in White men improvement of the lipoprotein-lipid profile was related to increased PH-LPL activity, no association between PH-LPL (or PH-HL) and lipoprotein-lipid variables was observed in Black men. Results of the present study suggest that in Whites, the increase in PH-LPL activity in response to endurance exercise training is associated with a better lipoprotein-lipid profile, therefore reducing CHD risk. However, the generally better metabolic profile of Black individuals may minimize further improvement of lipoprotein-lipid concentrations by exercise training. PMID- 11730821 TI - The associations among thoracic aortic atherosclerosis, coronary atherosclerosis and the function of high density lipoprotein. AB - We sought to determine the associations among thoracic aortic atherosclerosis, coronary atherosclerosis and the function of high density lipoprotein (HDL) in a case-control study. The function of HDL can be assessed by the fractional esterification rate of cholesterol in low density lipoprotein (LDL)- and very low density lipoprotein (VLDL)-depleted plasma (FER(HDL)), which reflects a balance of cholesterol uptake by HDL and cholesterol ester (CE) transport in the reverse cholesterol transport (RCT) system in humans. Cases (n=51, age: 64.3+/-8.0 years) and controls (n=51, age: 58.7+/-13.1 years) were defined as subjects with/without angiographically proven coronary artery disease (CAD), respectively and examined for thoracic aortic atherosclerosis (TAA) by transesophageal echocardiography. The severity of TAA was determined by the ratio of average sclerotic areas (ASA) and average sclerotic lengths (ASL). The cases had significantly (P<0.05) higher values of ASA (0.22+/-0.18 vs. 0.10+/-0.11), ASL (0.82+/-0.56 vs. 0.48+/-0.45), ASA/ASL ratio (0.23+/-0.08 vs. 0.17+/-0.09) and FER(HDL) (10.3+/-3.8 vs. 8.3+/ 3.5% per hour) and lower HDL-C and apolipoprotein A-I levels than the controls. A receiver operating characteristic (ROC) curve analysis showed that ASA/ASL and FER(HDL) had moderate discriminating ability for CAD and the diagnostic accuracy of ASA/ASL was better than that of FER(HDL) (area under ROC curve: 0.703 and 0.656, respectively). Multivariate logistic regression analysis indicated that ASA/ASL and FER(HDL) were independent indicators for CAD [odds ratio (95% CI): 7.5 (2.4-27), P<0.01 and 4.0 (1.2-15), P<0.05] after adjusting for age, gender and other conventional risk factors, and that a high FER(HDL) value greatly increased the relative risk of CAD associated with a high ASA/ASL. The function of HDL, as assessed by FER(HDL), enhances the ability of TAA to predict CAD. PMID- 11730822 TI - Association of the A/T54 polymorphism in the intestinal fatty acid binding protein with variations in plasma lipids in the Framingham Offspring Study. AB - We investigated the potential role of the genetic variation at the intestinal fatty acid binding protein gene (FABP2) in influencing lipid levels in a representative sample of the Framingham Offspring Study participants (n=1930). In men, the T54 allele was associated with significantly higher LDL-cholesterol (3.47+/-0.83 vs. 3.36+/-0.83 mmol/l; P<0.047), and ApoB (1.04+/-0.23 vs. 1.01+/ 0.24 g/l; P<0.020) after adjustment for familial relationship, age, BMI, smoking, alcohol intake and the use of beta-blockers compared with the A54 allele. This relationship with ApoB continued to be significant after adjustment for APOE genotype (P<0.034). In women, the T54 allele was associated with significantly higher total-cholesterol (5.32+/-1.01 vs. 5.17+/-0.98 mmol/l; P<0.049) and LDL cholesterol (3.31+/-0.93 vs. 3.18+/-0.85 mmol/l; P<0.023) after adjustment for covariates and menopausal status, estrogen therapy and APOE genotype. In men, the T54 allele was associated with significantly higher levels of small VLDL and lower levels of large HDL. Moreover, there was no significant relationship between FABP2 alleles and lipoprotein diameter or the prevalence of coronary heart disease in both genders. Our data are consistent with the T54 IFABP increasing the flux of lipids through the enterocyte leading to an increase in chylomicron secretion. PMID- 11730823 TI - Effects of androgen manipulation on postprandial triglyceridaemia, low-density lipoprotein particle size and lipoprotein(a) in men. AB - Although androgenic hormones decrease HDLC concentration, no direct evidence has linked them to atherosclerosis. The present study was undertaken to extend our ability to assess risk associated with androgen induced lipoprotein(Lp) changes by simultaneously gathering information about postprandial triglyceridaemia (PPT), LDL particle size, HDL and Lp(a) in men either taking exogenous androgens or with suppressed endogenous androgen concentrations. The experimental groups comprised nine male bodybuilders who self-administered anabolic-androgenic steroids (AAS) for a mean period of 6.5 weeks, and 10 healthy men whose testosterone concentration had been reversibly suppressed for 5 weeks using the GnRH agonist triptorelin (Decapeptyl; D-Trp-6-LHRH). A separate group receiving no hormonal treatment provided analytical control (n=7). Lipoprotein size was assessed by gradient gel electrophoresis categorisation (GGE), lipoprotein concentrations by immuno and enzymatic assays and PPT by a standardised oral fat tolerance test (65g /m(2)). Testosterone concentration was significantly reduced on triptorelin from 7.32+/-1.92 to 1.15+/-0.57 ng/ml (P=0.002). High dose AAS use was confirmed by urinalysis. With AAS use, mean HDLC and Lp(a) concentrations and PPT decreased from 0.9+/-0.3 to 0.7+/-0.3 mmol/l (P=0.004), 125+/-128 to 69+/-73 U/l (P=0.008) and 11.6+/-10.0 mmol/l h to 7.5+/-5.4 mmol/l h (P=0.027) respectively. Mean total cholesterol and LDLC were unchanged. LDL size was unchanged in six AAS users, decreased in one but remaining in the normal size range, and increased in two from small LDL to the normal range. Size changes in the latter two subjects were associated with 42 and 58% reductions in PPT respectively. In the triptorelin group, mean total cholesterol, HDLC and Lp(a) were increased from 4.8+/-0.8 mmol/l to 5.2+/-1.0 mmol/l (P=0.039), 1.1+/-0.2 to 1.4+/-0.3 mmol/l (P=0.002) and 278+/-149 to 377+/-222 U/l (P=0.004) respectively. Mean LDLC concentration and PPT were unchanged. LDL particle size increased in four, decreased in two, and was unchanged in four subjects. LDL size decreased in two and showed no change in the other five control subjects. Other lipid measures were unchanged in the control group. Thus, apart from lowering HDLC concentrations, no other potentially atherogenic effects of endogenous androgens or AAS were observed. A suppression of Lp(a) as well as a reduced PPT and increased LDL size in predisposed individuals may be antiatherogenic effects of AAS. PMID- 11730824 TI - Lowering of HDL cholesterol in post-menopausal women by tibolone is not associated with changes in cholesterol efflux capacity or paraoxonase activity. AB - Low HDL cholesterol increases the risk of coronary heart disease. Treatment of postmenopausal women with tibolone lowers HDL cholesterol. We elucidated the consequences of this unwanted side effect in a randomized, double-blind study, where 12 women received 2.5 mg tibolone per day and 6 women, placebo. Blood samples were collected on days -1 (i.e. baseline), 28, 56, and 84 for the analysis of various parameters of lipid metabolism and HDL function. Compared to placebo, treatment with tibolone led to statistically significant decreases of HDL cholesterol (-22% to -32%), apoA-I (-14% to -22%), and HDL subclass LpA-I ( 30% to -40%) but to no significant changes in apoA-II and HDL subclass LpA-I,A II. These changes were not associated with statistically significant changes in the activity of plasma to release 3H-cholesterol from radiolabeled fibroblasts or in the serum activity of the anti-oxidative enzyme paraoxonase/arylesterase. There were no significant changes in either serum levels of triglycerides, LDL cholesterol, apoB, and leptin, or in LDL size. We conclude that changes in insulin do not contribute to the lowering of HDL cholesterol by tibolone. Despite decreased HDL cholesterol, putatively anti-atherogenic activities of HDL remained unchanged. PMID- 11730825 TI - Postprandial lipid metabolism and insulin sensitivity in young Northern Europeans, South Asians and Latin Americans in the UK. AB - The aim of this study was to determine whether the higher susceptibility to coronary heart disease and diabetes in South Asian immigrants to the United Kingdom compared with the Caucasian population may reflect insulin resistance and altered postprandial lipid metabolism. We also wished to study Latin Americans, an ethnic group that has not been previously studied in the United Kingdom. The intention was to carry out a detailed study in a relatively small number of subjects to provide essential baseline information for larger epidemiological studies. Postprandial lipaemia was measured in 25 subjects (eight South Asians, eight Latin Americans and nine Northern Europeans) who resided in the United Kingdom. Results from the postprandial studies were correlated to insulin sensitivity measured by the insulin tolerance test, food intake, anthropometry and adipose tissue fatty acid composition. In South Asians, postprandial glucose and insulin concentrations were greater than in the other groups (P<0.01 and <0.05, respectively), although there were no differences in postprandial triacylglycerol concentrations or in insulin sensitivity assessed with the insulin tolerance test. The decreased glucose tolerance in this group could not be explained by differences in percentage body fat, body mass index, or by dietary intake measured by food records or adipose tissue fatty acid composition. We conclude that postprandial lipaemia is not affected in young South Asians compared to Northern Europeans although glucose intolerance is detectable. The results from the present study should help in the design of further postprandial lipid studies in different ethnic groups. PMID- 11730826 TI - Influence of CETP gene variation on plasma lipid levels and coronary heart disease: a survey in Taiwan. AB - Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester from high density lipoprotein (HDL) to very low-density lipoprotein (VLDL), low-density lipoprotein (LDL) and chylomicron in exchange for triglycerides. Two CETP genetic variation and four polymorphisms are investigated by polymerase chain reaction (PCR) and restriction enzyme digestion in a population of Taiwan. The results show that a very rare variation frequency is found for CETP intron 14 splice site G-->A change. The population shows a predominant 405Ile allele (61%), 442Asp (97.7%), intron 1Taq1B(+) G allele (52%), intron 8 Msp1(-) A allele (89%) and intron 9 EcoN1(-) T allele (59.2%) in the control group. Patients with coronary heart disease (CHD) have more CETP EcoN1(+/+) GG genotype (25.3%) than the controls (13.6%) (P=0.049). The intron 1 Taq1B(-) A allele is associated with a high HDL cholesterol and apoA1 level, the EcoN1(+) G allele with a low apoA1 level and the 442Gly with both high total cholesterol and LDL cholesterol levels. Paradoxically, the 442Gly is also present with a higher frequency (5.2%) in HDL cholesterol > or =65 mg/dl group than that in the general population (2.3%) (P=0.04). PMID- 11730827 TI - Effects of a high polyunsaturated fat diet and vitamin E supplementation on high density lipoprotein oxidation in humans. AB - Oxidative modification of high-density lipoproteins (HDL) impairs several biologic functions critical to its role in reverse cholesterol transport. We therefore investigated the effect of dietary polyunsaturated fat and vitamin E on the kinetics of HDL oxidation. Ten subjects were fed sequentially: a baseline diet in which the major fat source was olive oil; a high polyunsaturated fat diet in which the major fat source was safflower oil; and the safflower oil diet plus 800 I.U. vitamin E per day. Plasma lipoprotein levels, vitamin E content, fatty acid composition, and oxidation lag time and rate were determined after 3 weeks on each diet. The polyunsaturated fat diet increased the mean HDL(2) lag time from 45.8+/-12.5 to 83.3+/-11.6 min with no change in oxidation rate. Addition of vitamin E further increased the HDL(2) lag time to 115.6+/-4.4 min and decreased the HDL(2) oxidation rate 10-fold. Neither the polyunsaturated diet alone nor the diet with vitamin E supplementation had any effect on HDL(3) oxidation. We conclude that under conditions of controlled dietary fat intake, a high polyunsaturated fat intake does not increase the oxidation susceptibility of HDL subfractions, and that in this setting, vitamin E supplementation reduces the oxidation susceptibility of HDL(2). These data suggest that antioxidants could influence HDL function in vivo. PMID- 11730828 TI - The effect of continuous combined hormone replacement therapy on arterial reactivity in postmenopausal women with established angina pectoris. AB - Most epidemiological studies have suggested that the administration of estrogen reduces cardiovascular risk in healthy postmenopausal women. More recently, however, in the large Heart Estrogen/progestin Replacement Study (HERS), it was unexpectedly found that in women with established cardiovascular disease, there was overall no difference in cardiovascular events between those treated with combined oestrogen/progestin hormone replacement therapy and those on placebo. The aim of this study was to examine the effect of combined hormone replacement therapy on arterial reactivity in women with existing angina pectoris. Seventy four postmenopausal women with angina pectoris were recruited into a 16 week double-blind, placebo-controlled study of treatment with 2 mg of estradiol combined with 1 mg of norethisterone acetate daily. The median endothelium dependent change in arterial relaxation increased from 5.00 to 7.69% in the treatment group and decreased from 5.57 to 3.64% in the controls. The median endothelium-independent change in arterial relaxation increased from 6.49 to 7.27% in the treatment group and decreased from 4.39 to 2.07% in the controls. The changes in arterial relaxation between the treatment and control groups were not statistically significant. The administration of estrogen/progestin did not significantly improve either endothelium-dependent or -independent arterial relaxation in postmenopausal women with established cardiovascular disease. We have previously shown that estrogen/progestin treatment improves endothelium dependent relaxation in healthy women. The results of our study provide one possible explanation for the clinical findings of the HERS study. In women with established cardiovascular disease, arterial relaxation does not increase significantly in response to treatment with combined hormone replacement therapy. PMID- 11730829 TI - Lipoprotein and apolipoprotein abnormalities in familial combined hyperlipidemia: a 20-year prospective study. AB - In order to characterize the lipoprotein abnormalities in familial combined hyperlipidemia (FCHL) and to describe factors associated with the stability of the FCHL phenotype during 20-year follow-up, 287 individuals from 48 families with FCHL originally identified in the early 1970s (baseline) were studied. Hyperlipidemia was defined as lipid-lowering medication use, or > or =age- and sex-specific 90th percentile for triglycerides or cholesterol. Triglyceride, cholesterol and medical history data were obtained at baseline and 20-year follow up. Additional follow-up measures included HDL-C, LDL-C, LDL particle size, lipoprotein(a), apolipoprotein (apo) A-I, apoB, and apoE polymorphism. Longitudinally, two-thirds of relatives were consistently normolipidemic or hyperlipidemic, and one third were discordant for hyperlipidemic status at baseline and 20-year follow-up. Individuals with hyperlipidemia at baseline and/or follow-up had higher apoB levels than those with consistently normal lipids (P<0.05), whereas small LDL size was associated with concurrent hyperlipidemia. Among individuals who were normolipidemic at baseline, the following variables were independently associated with development of hyperlipidemia over 20 years: older age at baseline, male sex, greater increase in BMI during follow-up, and apoE alleles epsilon 2 or epsilon 4. In conclusion, apoB is associated with hyperlipidemia and apoE polymorphism is associated with later onset of hyperlipidemia in FCHL. PMID- 11730830 TI - Association between self-reported physical activity and vascular reactivity in postmenopausal women. AB - Short-term exercise training has been associated with improved endothelial dependent vasodilation, but the impact of long-term habitual physical activity on vascular reactivity is not established. We studied the correlation between self reported, habitual physical activity and vasoreactivity in non-smoking, non diabetic, postmenopausal women (n=34, mean age 65.6+/-7.4 years). Vasoreactivity was evaluated by the percentage and absolute change in brachial artery diameter in response to reactive hyperemia induced by occlusion-release, and in response to cold pressor testing (CPT). Habitual physical activity was assessed by a standardized questionnaire based on participant recall. Our results indicate that 64.7% of the women were exercising-to-sweat > or =1x/week, 4.8 flights of stairs were climbed/day, 5.0 city blocks were walked/day and 29.4% participated in moderately physically demanding daily activity. There was a significant association between the number of city blocks walked daily and exercising-to sweat > or =1x/week with brachial artery percentage and absolute change to CPT (P<0.05). Women who reported a moderately physically demanding daily activity had a significantly greater brachial reactivity percentage change in response to CPT compared with those performing less demanding daily activity (2.0+/-3.6 versus 1.4+/-7.0%, P<0.05). The response to reactive hyperemia was also greater in those women reporting moderately physically demanding daily activity compared to less active women (6.5+/-5.4 versus 5.8+/-5.9%, P=n.s.), but this did not reach statistical significance. Stepwise, multivariate analysis adjusting for body mass index and HDL-cholesterol eliminated the association between physical activity and brachial reactivity in response to CPT, suggesting that physical activity may affect vasoreactivity via these mechanisms. This study suggests that moderate levels of self-reported physical activity are associated with a greater brachial reactivity in response to CPT and supports the recommendation that moderate intensity physical activity may be cardioprotective in postmenopausal women. PMID- 11730831 TI - Retinoic acid suppresses endothelin-1 gene expression at the transcription level in endothelial cells. AB - Retinoids have been shown to inhibit cell growth, which can result in an anti atherosclerotic action in the vasculature. Endothelin-1 (ET-1), a potent vasoconstrictor peptide produced in endothelial cells, plays an important role in inducing proliferation of vascular smooth muscle cells. In this study, we investigated the effect of retinoids on the mRNA expression and transcriptional activity of the ET-1 gene in endothelial cells. All-trans retinoic acid (ATRA) suppressed ET-1 mRNA expression in cultured endothelial cells. Synthetic retinoids, Ch55 and Am580 (retinoic acid receptor (RAR) agonists) markedly enhanced this effect, and an RAR antagonist, LE540, blocked this inhibitory effect on ET-1 gene expression. ATRA did not change ET-1 mRNA half-life. Transfection experiments using 5 kb of the ET-1 promoter-reporter gene construct which contains 5 kb of the preproET-1 promoter revealed that ATRA and Ch55 suppressed ET-1 promoter activity, resulting in down-regulation of ET-1 gene transcription. Taken together, retinoids may be another modulator of endothelial cell function through regulation of vasoactive substances at the transcription level. PMID- 11730832 TI - Homocysteine and cardiovascular disease in diabetes mellitus. AB - BACKGROUND: Patients with diabetes mellitus (DM) have 2- to 6-fold increase in the prevalence of cardiovascular disease (CVD) compared to non-DM subjects. Epidemiological data show that DM is synergic with other conventional risk factors. Total plasma homocysteine (tHcy) is an emerging CVD risk factor. We reviewed the literature to explore the relation between tHcy and CVD in patients with DM. METHODS: We searched the MEDLINE database for articles on homocysteine, DM and CVD published from January 1991 to October 2000. RESULTS: The mean plasma tHcy level is usually low or normal in DM patients, except when nephropathy is present. Levels in that case tend to be higher than in non-DM patients. An independent association with tHcy and CVD was shown in retrospective studies, for DM patients. Prospective studies showed an association between elevated tHcy and all cause mortality in DM patients. In general, the association between elevated levels of tHcy and the outcome was stronger than in non-DM individuals, for all types of study. DISCUSSION: To date, there are no prospective work that specifically examined the relationship between levels of tHcy and the presence of CVD in the DM population. Nor are there studies to show that treating elevated tHcy results in a reduction of CVD events. Such studies are ongoing. Nevertheless, since hyperhomocysteinemia is potentially reversible with vitamin therapy, interaction of DM with high levels tHcy on the risk of CVD may have consequences with regard to management of primary and secondary prevention in DM patients who are at particularly high risk of CVD events. PMID- 11730833 TI - Atorvastatin improves blood rheology in patients with familial hypercholesterolemia (FH) on long-term LDL apheresis treatment. AB - To determine the effect of atorvastatin on blood rheology in patients with familial hypercholesterolemia (FH) on regular LDL apheresis, we prospectively studied the rheological variables fibrinogen, plasma viscosity, red cell aggregation, whole blood viscosity, hematocrit and platelet aggregation in 12 patients (two homozygous, ten heterozygous) before and during treatment with atorvastatin. Baseline values of red cell aggregation and whole blood viscosity were increased in FH patients on regular LDL apheresis compared with healthy controls (P<0.05), whereas fibrinogen, plasma viscosity and hematocrit were similar in the two groups. Treatment with atorvastatin reduced red cell aggregation (P<0.01), whole blood viscosity (P<0.01), plasma viscosity (P<0.01) and platelet aggregation (P<0.05), but caused a slight increase in plasma fibrinogen (by 5%; P<0.01). Our findings suggest that atorvastatin improves blood rheology in patients with FH on regular LDL-apheresis. This improvement in blood flow properties may contribute to the well-known beneficial effects of atorvastatin on cardiovascular risk in patients with severe hyperlipidemia and atherosclerotic vascular disease. PMID- 11730834 TI - Prevalence of Chlamydia pneumoniae in peripheral blood mononuclear cells in Italian patients with acute ischaemic heart disease. AB - Chlamydia pneumoniae infection generally starts in the respiratory tract and probably disseminates systemically in the blood stream within alveolar macrophages. We investigated the prevalence of C. pneumoniae DNA in peripheral blood mononuclear cells (PBMC) in patients with acute ischaemic heart disease. Samples of blood were obtained from 93 consecutive patients with acute ischaemic heart disease and from 42 healthy subjects, for detection of C. pneumoniae DNA in PBMC by polymerase chain reaction (PCR) and for serology. C. pneumoniae DNA in PBMC was detected in 25.8% (24/93) of the patients with acute ischaemic heart disease and in 4.8% (2/42) of the healthy subjects (P=0.008). C. pneumoniae IgG was found in 76.3% of patients and in 45.2% of healthy subjects (P=0.0008) while C. pneumoniae IgA was found in 59.1% and in 33.3%, respectively (P=0.01). No correlation was found between anti-C. pneumoniae antibody titers and positive PCR results. The detection of C. pneumoniae DNA in PBMC may aid in selecting patients who may benefit from antibiotic treatment; however, to support this contention, longitudinal studies on patients treated with antibiotics would also be necessary. PMID- 11730835 TI - Beneficial effect of quinapril in patients with angiotensin-converting enzyme D allele after coronary stenting. PMID- 11730836 TI - Identification of novel immunogenic proteins of Helicobacter pylori by proteome technology. AB - Cell surface proteins of the human gastric pathogen Helicobacter pylori, reference strain CCUG 17874, were extracted with acid glycine and fractionated by heparin affinity chromatography. The extracts were subsequently analysed using high-resolution two-dimensional gel electrophoresis (2-DE) and immunoblotting. Four proteins of low molecular masses (25-30 kDa) stained by Coomassie R-350, were identified by peptide ESI-MS/MS sequencing after in-gel tryptic digestion. The identified proteins were recognised by sera from H. pylori-infected patients. Two of them are now described for the first time as immunogenic proteins of which one protein was determined to be distinct from all H. pylori proteins previously described. In addition, the specificity of the identified peptides was evaluated using both 1-D and 2-D immunoblotting against a panel of sera from patients with various bacterial infections. The present identification of highly specific antigens of H. pylori will encourage the improvement of serological diagnostic tests to diagnose and monitor H. pylori infection. PMID- 11730837 TI - Evaluation of total IGF-I assay methods using samples from Type I and Type II diabetic patients. AB - Measurements of circulating insulin-like growth factor-I (IGF-I) levels are an important part of many studies on growth and development. Circulating IGF-I levels are growth hormone (GH) dependent and are also impacted by age, gender, nutritional status and disease. Moreover, IGF-I is the main pharmacodynamic marker of GH activity. The majority of circulating IGF-I is associated with high affinity insulin-like growth factor-binding proteins (IGFBPs), making accurate and precise measurements of total IGF-I concentrations in biological matrices technically challenging. Many total IGF-I assay methods combine an immunoassay with a sample preparation method aimed at removing IGFBPs. However, not all sample preparation methods efficiently remove all IGFBPs or BP fragments (BPFs), and there is currently no reference method for IGF-I measurement against which these IGF-I assays can be calibrated. We have evaluated a number of IGF-I immunoassays and sample preparation methods using plasma samples from normal donors and from donors with Type I and Type II diabetes mellitus. In order to eliminate the variability between assays due to differences in assay standardization, we used the same preparation of highly pure, fully characterized IGF-I as the standard for all assays. We found that the data produced by many of the IGF-I assay methods showed good agreement when IGF-I levels in samples from normal individuals were measured. However, we found that these agreements were quite poor when IGF-I levels in samples from diabetics were measured. This was true of methods that claimed to physically separate IGFBPs from IGF-I either by acid/ethanol extraction or by acid chromatography. Several methods have recently been developed that physically separate IGF-I from IGFBPs followed by a chemical displacer to displace any residual BPs or BPFs from IGF-I. We found that the data generated by these displacement methods showed good agreement when assaying samples from diabetic as well as normal donors. There is considerable discussion in the literature as to whether individuals with diabetes have normal circulating levels of IGF-I. Many of the published studies are based on assays that may not accurately measure IGF-I levels due to problems with assay standardization and/or with assay methodology. Displacement methods may enable us to more accurately measure IGF-I levels in diabetes. PMID- 11730838 TI - Analysis of transforming growth factor-beta 1 (TGF-beta1) expression in human monocytes infected with Mycobacterium avium at a single cell level by ELISPOT assay. AB - Transforming growth factor beta 1 (TGF-beta1) has been implicated in the pathogenesis of a number of diseases including infection with intracellular pathogens such as Mycobacterium avium complex (MAC). In this study, we developed an ELISPOT assay for measurement of active TGF-beta1 produced by peripheral blood mononuclear cells (PBMC) from healthy individuals in response to LPS or MAC. The frequency of TGF-beta1 producing cells was significantly (p<0.04) higher in response to LPS (10 microg/ml) as compared to unstimulated cells (n=4). Moreover, the frequency of TGF-beta1 producing cells was threefold higher in monocyte (MN) enriched cell population than those in PBMC indicating that the source of TGF beta1 producing cells in PBMC was MN. In addition, the frequency of TGF-beta1 producing cells in response to MAC (10:1, cfu:MN) was significantly higher (p<0.03) than unstimulated cells. However, the frequency of TGF-beta1 producing cells in response to MAC (10:1) was eight to ninefold lower than that by LPS (10 microg/ml). Moreover, there was a correlation between the level of total TGF beta1 in 24-h culture supernatants and the number of TGF-beta1 producing cells upon MAC stimulation. TGF-beta1 ELISPOT-assay may be a sensitive and a powerful tool for detection of TGF-beta1 producing cells, and may be helpful in elucidation of the nature of TGF-beta1 production at sites of diseases. PMID- 11730839 TI - Quantitative measurement of anti-ErbB-2 antibody by flow cytometry and ELISA. AB - To establish standard methods for measuring anti-ErbB-2 antibody, a flow cytometric and an enzyme-linked immunosorbent assay (ELISA) were developed and compared. In the flow cytometric assay, the antibody was measured by binding to human breast cancer cell line SKBR3 and the result expressed as mean channel fluorescence (MCF). In ELISA, the antibody was measured by binding to a recombinant, secreted human ErbB-2 containing the N-terminal 505 amino acids of ErbB-2 fused to myc and His tags (secE2/myc/His or secE2) and the result expressed as O.D.(405). A mouse anti-human ErbB-2 mAb 9G6 was used as the standard. Using flow cytometry, MCF of 9G6 binding increased with linearity between 0.6 and 10 microg/ml. Using ELISA, O.D.(405) increased with linearity between 0.015 and 1 microg/ml, indicating greater sensitivity of ELISA. The titer of an immune mouse serum was determined to be 400 and 8000 with flow cytometric and ELISA assay, respectively, consistent with the assay sensitivity. Based on standard curves generated with 9G6, antibody concentration in the same serum sample was calculated to be approximately 230 microg/ml by ELISA and approximately 270 microg/ml by flow cytometry. Therefore, excellent concordance in antibody concentration was obtained with different assays using linear regression and properly diluted samples. This concordance was verified with multiple serum samples. PMID- 11730840 TI - Selection of high affinity p-azophenyarsonate Fabs from heavy-chain CDR2 insertion libraries. AB - The length of the heavy chain complementarity-determining region two (HCDR2) of the unmutated anti-p-azophenylarsonate (Ars) monoclonal antibody (36-65 mAb) was extended by three residues in order to test whether this insertion can provide additional contacts between the Ab and the antigen. Two libraries were generated using 36-65 heavy and light chain genes which were cloned as Fab in the phage display vector pComb3. In the first library, three randomized amino acids were inserted between residues Gly 54 and Asn 55, which are the most solvent exposed residues in the HCDR2 loop. In the second library, in addition to the 3-mer randomized insertion, the flanking residues at positions 54 and 55 were also randomized to allow additional loop flexibility for binding to Ars. Solid-phase and solution phase affinity panning were used to select for clones that bind to Ars. Results indicate that diverse 3-mer HCDR2 insertions can be tolerated, and affinities 10-fold higher than germline encoded 36-65 Ab can be obtained. The sequence diversity of the insertion among the selected clones from both libraries suggests that the insertion increases contact between the Ab and the protein carrier rather than the hapten alone. PMID- 11730841 TI - Cytokine mRNA quantification by real-time PCR. AB - Real-time PCR represents a new methodology that accurately quantifies nucleic acids. This has been made possible by the use of fluorogenic probes, which are presented in two forms, namely hydrolysis probes (also called TaqMan probes) and hybridisation probes. We decided to apply this methodology to cytokine mRNA quantification and this led us to the development of a protocol that provides an easy way to develop and perform rapidly real-time PCR on a Lightcycler instrument. It was made possible by the use of freely available software that permits a choice of both the hydrolysis probe and the primers. We firstly demonstrated that the reproducibility of the method using hydrolysis probes compares favourably with that obtained with hybridisation probes. We then applied this technique to determine the kinetics of IL-1ra, IL-1beta, IL-5, IL-13, TNF alpha and IFN-gamma induction upon stimulation of human peripheral blood mononuclear cells (PBMC) by phytohaemagglutinin (PHA). Finally, the method was also used successfully to demonstrate that IFN-alpha induces IL-10 mRNA accumulation in human monocytes. PMID- 11730842 TI - Detection of anti-p53 antibodies by ELISA using p53 synthetic or phage-displayed peptides. AB - Anti-p53 antibodies have been detected in the sera of patients with various types of cancers. In this report, we describe the development of a new ELISA aimed at detecting anti-p53 antibodies using two peptides belonging to immunodominant epitopes of the p53 N-terminal region. We first tested the reactivity of the sera by an indirect ELISA using the peptides as a capture system. Then, the specificity of the reaction was confirmed by an inhibition assay. Two systems of peptide presentation, phage display and the streptavidin/biotin system, were evaluated. Using a panel of sera from cancer patients, both systems were found to be equally reliable, demonstrating that both peptide-based ELISAs can be used for the specific detection of anti-p53 antibodies. The presence of anti-p53 antibodies was associated with p53 alteration whether it be mutation or accumulation. PMID- 11730843 TI - Comparative properties of two peptide-antibody interactions as deduced from epitope delineation. AB - The linear epitope recognized by three closely related antibodies specific for the E6 oncoprotein of papillomavirus type 16 was delineated by phage display, spot peptide synthesis on cellulose membranes, and kinetic measurements with antigenic variants using a BIACORE. The same approaches, recently applied to an antibody specific for tobacco mosaic virus protein, led to the clear-cut delineation of a functional epitope comprising four key positions with well defined physico-chemical properties. In contrast, the E6 system is characterized by a non-essential contribution to binding of various factors, so that combinations of alternative properties are compatible with measurable binding activity. PMID- 11730845 TI - The use of HLA-A*0201-transfected K562 as standard antigen-presenting cells for CD8(+) T lymphocytes in IFN-gamma ELISPOT assays. AB - ELISPOT assays are increasingly used for a direct detection and quantification of single antigen-specific T cells in freshly isolated peripheral blood mononuclear cells (PBMC). They are particularly attractive for the monitoring of specific T lymphocyte responses in clinical trials assessing antigen-specific immunizations in patients with cancer or chronic viral infections. However, one major limitation for the broad clinical implementation of ELISPOT assays is the lack of an inexhaustible source of suitable HLA-matched antigen-presenting cells (APC). Currently available allogeneic or xenogeneic APC (such as the human lymphoid hybrid T2 or HLA-transfected insect cells) can either lead to strong background spot production by APC-reactive T lymphocytes or have a low antigen presentation capability. Both phenomena can prevent the detection of low frequency T cell responses in PBMC. In search of alternative APC for ELISPOT assays, the human chronic myelogenous leukemia cell line K562 that per se does not express HLA class I and class II molecules on the cell surface was transfected with the HLA A*0201 gene. Clonal HLA-A*0201-expressing K562 (K562/A*0201) cells were able to process and present endogenously expressed and exogenously loaded melanoma peptide antigens to HLA-A*0201-restricted cytolytic T lymphocyte clones in cytotoxicity and IFN-gamma ELISPOT assays. K562/A*0201 cells were then used as APC in IFN-gamma spot assays to detect ex vivo CD8(+) T lymphocytes responsive to known HLA-A*0201-binding peptide epitopes derived from cytomegalovirus, Epstein Barr virus, influenza virus and melanoma in PBMC from HLA-A*0201-positive donors. In the majority of cases, peptide-pulsed K562/A*0201 cells were similarly efficient in the ability to visualize single antigen-specific CD8(+) T lymphocytes when compared to T2 cells. However, in contrast to T2, background reactivity of CD8(+) T cells responsive to unpulsed K562/A*0201 was regularly found to be negligible, thereby enhancing the sensitivity of the ELISPOT assay, particularly in donors with strong anti-T2 reactivity. K562 cells transfected with HLA-A*0201 or other HLA genes can serve as standard APC for monitoring T lymphocyte responses against tumor and viral peptide antigens. PMID- 11730844 TI - Biodegradable poly (DL-lactide glycolide) microparticles as a vehicle for allergen-specific vaccines: a study performed with Ole e 1, the main allergen of olive pollen. AB - Ole e 1, the main allergen of the olive pollen, was entrapped in poly (DL-lactide glycolide) microparticles by a solvent evaporation double emulsion technique. The physical properties of the microparticles, such as particle size, surface morphology, allergen entrapment rate and antigen release pattern were investigated. Microparticles with a spherical morphology displayed a size range of less than 2 microm in diameter and antigen loading up to 60-70% (w/w). SDS PAGE and immunoblotting of the released Ole e 1 confirmed that the molecular integrity and the antigenic properties of Ole e 1 remained unaltered by the encapsulation process or polymer degradation. This finding suggests that microparticles displaying small particle sizes, rapid antigen release and a high allergen/polymer ratio may be a suitable delivery system for antigen in hyposensitization therapy against allergy. PMID- 11730846 TI - Assessment of positivity in immuno-assays with variability in background measurements: a new approach applied to the antibody response to Plasmodium falciparum MSP2. AB - Measurements of immune responses often exhibit considerable heterogeneity, making it impossible to clearly distinguish responders and nonresponders to particular antigens. Typically, in, for example, enzyme-linked immunosorbent assay (ELISA) procedures, a nonexposed control group is used to assign a cutoff value of positivity, calculated as the mean plus either 2 or 3 standard deviations (S.D.). This can cause extremely biased estimates of response rates when the background is variable, and especially when there is overlap between the distribution of the control levels and that of responders. This problem is compounded when results of assays with different background levels are compared. We illustrate this with hypothetical data sets reflecting frequent patterns seen in laboratory and epidemiological studies. We propose that such data should be analysed by statistical modelling of the ratio of numbers of test samples/control samples as a function of the readout from the assay. Rather than classifying samples dichotomously as negative or positive, this provides estimates of the prevalence of positivity lambda, and the probability, for each sample, that the measured activity is above background. Several statistical methods can provide such estimates. Analyses of simulated data sets using our preferred estimation method [a latent class model (LCM)] demonstrate that this gives more reliable results than the traditional assignment using cutoff values. We have applied this approach to the analysis of ELISA assessments of antibodies against distinct regions of the Plasmodium falciparum merozoite surface protein 2 (MSP2) in human sera from Tanzania. PMID- 11730847 TI - Development, characterization and use of monoclonal antibodies against sTRAIL: measurement of sTRAIL by ELISA. AB - Two monoclonal antibodies against tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL), designated VI10E and III6F, have been generated. These antibodies were useful in flow cytometry analysis, immunohistochemistry, immunoprecipitation and in the development of an immunoassay for the detection of soluble TRAIL (sTRAIL)in biological samples. The immunoassay was based on two monoclonal antibodies against TRAIL. VI10E was used as the capture antibody and bound TRAIL was detected with anti-TRAIL from R&D Systems which was digoxigenin (DIG)-labeled. This enzyme-linked immunosorbent assay (ELISA) was specific for TRAIL since a panel of other cytokines did not affect the signal. The immunoassay was suitable for the detection of sTRAIL in human serum and plasma samples, cell culture supernatants and cell lysates. In a preliminary screening, it was found that serum samples from human immunodeficiency virus (HIV)-infected patients contained sTRAIL, and all these positive samples were found in the AIDS group. Using the immunoassay, it was found that phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) to produce significant amounts of sTRAIL, the levels of which increased with exposure time. Thus, the immunoassay for TRAIL presented here represents a useful tool for measuring sTRAIL in various biological samples. It will also permit studies of release mechanisms as well as possible functions of the soluble form of this molecule. PMID- 11730848 TI - Analysis of cellular immune responses in the peripheral blood of mice using real time RT-PCR. AB - Murine cancer models are commonly used in the evaluation of immunotherapeutic strategies. However, one of the major limitations in the monitoring of cellular immune responses induced by various vaccination approaches is that existing immunoassays require sacrifice of the animals for collection of the spleen or lymph nodes for analysis. We report here the development of an assay to quantitate antigen-specific T cell responses in murine blood, without euthanasia, using real-time RT-PCR for measurement of interferon-gamma mRNA levels. C57BL/6 mice were immunized with an adenoviral vector encoding the melanoma antigen gp100 (Ad2/gp100) or were left untreated. Small samples of whole blood were collected by retro-orbital puncture for analysis of T cell reactivity. The mice were then euthanized and spleen cells were isolated for comparative analyses. Blood and spleen cells were restimulated with either a peptide containing the dominant gp100 MHC Class I-restricted epitope, gp100(25-33), or a negative control peptide containing an irrelevant Class I-restricted epitope from ovalbumin. IFN-gamma mRNA was detected in gp100 peptide-pulsed whole blood as well as in spleen cells recovered from Ad2/gp100-treated mice, but not in untreated mice. In addition, there was a strong correlation in the magnitude of the gp100-specific response of spleen cells from an individual animal when measured by real-time RT-PCR with the more conventional enzyme-linked immunospot (ELISPOT) method (P<0.001). Finally, the gp100-specific immune response measured in the peripheral blood of individual animals by real-time RT-PCR or ELISPOT showed a significant correlation with the response measured in the spleen (P=0.001). We conclude that real-time RT-PCR measurement of IFN-gamma mRNA induced by antigenic stimulation is an attractive method to measure an antigen-specific cellular immune response in small samples of whole blood as it does not require euthanasia, mirrors the response observed in the spleen and correlates with the response measured using the conventional ELISPOT method. PMID- 11730849 TI - Generation of heterohybridomas secreting human immunoglobulins; pokeweed mitogen prestimulation is highly effective but phytohemagglutinin drives most B cells into apoptosis. AB - Human monoclonal antibodies have commonly been generated by forming hybridomas of stable lymphoblastoid cell lines and Epstein-Barr virus (EBV)-transformed human B cells that have been exposed to phytohaemagglutin (PHA)-stimulated T cells. However, this technique has predominantly given rise to IgM- but very rarely IgG- or IgA-producing clones. We now report that, regardless of prior EBV infection, pokeweed mitogen (PWM) stimulation of human peripheral blood mononuclear cells (PBMCs) generated much higher numbers of IgM-, IgA- and IgG-producing B cells than did stimulation with PHA. Fusion of PWM-stimulated PBMCs with a mouse myeloma cell line also gave rise to 7- to 12-fold higher numbers of IgG- and IgA producing heterohybridomas than PBMCs that were prestimulated with PHA. Judged by Annexin V staining, stimulation with PHA induced a very high rate of B cell apoptosis within 24 h, whereas, even after 7 days, PWM stimulation only induced marginal B cell apoptosis. This should explain why PHA is much inferior to PWM in stimulating immunoglobulin (Ig) production in vitro and in generating immunoglobulin-producing human B cell hybridomas. It is concluded that PWM stimulation may greatly facilitate the generation of human monoclonal antibodies of all isotypes. PMID- 11730850 TI - Generation of neutralizing mouse anti-mouse IL-18 antibodies for inhibition of inflammatory responses in vivo. AB - The proinflammatory cytokine IL-18 mediates IFN-gamma production as well as the induction of Th1 polarized immune responses in synergy with IL-12. In this study, we describe the production of isogeneic monoclonal antibodies (Mabs) directed against murine IL-18 (mIL-18). Immunization of IL-18-deficient mice with recombinant mIL-18 in the presence of CpG-oligodeoxynucleotides (CpG-ODN) and alum as adjuvant resulted in high anti-IL-18 serum titers. We could identify two Mabs, SK721-2 and SK113AE-4, which were able to bind to IL-18 and neutralize its IFN-gamma inducing effect in vitro with an IC(50) of 40-100 ng/ml. In vivo, LPS induced IFN-gamma production was reduced by 60-85% following a single administration of Mabs SK113AE-4 or SK721-2. Since IL-18 is likely to be involved in the pathogenesis of inflammatory diseases such as rheumatoid arthritis or Crohn's disease, neutralizing mouse anti-mouse IL-18 Mabs have the potential to become valuable tools for the therapeutic exploration of long-term IL-18 blockade in vivo. PMID- 11730851 TI - An improved flow cytometric assay for the determination of cytotoxic T lymphocyte activity. AB - The cytotoxic activity of T lymphocytes, natural killer and lymphokine-activated killer cells is usually tested by radioactive assays, which detect the release of cytoplasmic contents after plasma membrane disintegration of dying cells. In contrast to this indirect evaluation of cytotoxicity, we describe here an improved fluorescence assay that is based on the direct quantitative and qualitative flow cytometric analysis of cell damage at a single cell level. Target cells are stained with PKH-26, a lipophilic dye that stably integrates into the cell membrane and permits distinction between target and effector cells. After 3 h of in vitro incubation, costaining with AnnexinV-FITC (ann-FITC) and propidium iodide (PI) permitted discrimination between vital, early apoptotic and necrotic cells. Data analysis is performed first by gating on PKH-26-positive target cells followed by the analysis of ann-FITC- and PI-positive subpopulations. The percentage of cytotoxicity in the PKH-26-gated cell population is calculated by subtracting non-specific ann-FITC- or PI-positive target cells, measured in appropriate controls without effector cells. Membrane staining of target cells such as primary melanoma cells or leukemic blasts revealed high and stable loading of PKH-26 without altering the viability or the immunogenicity of the cells. Using in vitro-generated antigen-specific cytotoxic T lymphocytes (CTL), we could demonstrate that this flow cytometric assay is sensitive and correlates well with the standard 51Cr release assay. In conclusion, the improved fluorescence assay is a simple and highly reproducible procedure for evaluating the specific cytotoxicity of T cells. PMID- 11730852 TI - Phenotypic characterization of porcine IFN-gamma-producing lymphocytes by flow cytometry. AB - We have developed a three-colour flow cytometric assay for phenotypic characterization of porcine IFN-gamma-producing lymphocytes. Analyses of activated swine peripheral blood mononuclear cells (PBMC) showed a significant difference in the proportion of IFN-gamma producing cells between young and adult animals (13.2+/-5.8% versus 34.2+/-5.7%). The majority of IFN-gamma producing cells were alphabeta T lymphocytes, although there was also an important proportion of gammadelta T cells particularly in young animals. Within the alphabeta T lymphocytes, the double positive CD4(+)CD8(lo) subset, that contains memory T cells, produced high levels of IFN-gamma, whereas the CD8(hi) T cells ranged from low to high levels of IFN-gamma. Also, consistent with a higher production by memory T cells, the CD45RA(-) subset of both CD4(+) and CD8(+) cells contained higher numbers of IFN-gamma producing cells than the CD45RA(+) subset. Finally, no production of IFN-gamma by either B cells (CD21(+)) or monocytes (SWC3(+)) was detected. This assay may be useful for the assessment of cell-mediated immunity in vaccine trials and may contribute to our understanding of the role of IFN-gamma in protective immunity against important viral diseases of the pig. PMID- 11730853 TI - CD40 ligand and lipopolysaccharide enhance the in vitro generation of melanoma reactive T-cells. AB - Cancer vaccine trials require sensitive assays for evaluating T-cell responses in immunized patients. In addition, these methods are used for identifying novel tumor-associated antigens (TAA). Therefore, our aim was to improve the methods for evaluating patients receiving the cancer vaccines by enhancing the in vitro detection of tumor-specific T cells from the peripheral blood. We have developed an efficient and reproducible method for detecting tumor-specific T cells by optimizing the activation of antigen presenting cells (APC) in peripheral blood mononuclear cells (PBMC) of metastatic melanoma patients with soluble trimeric CD40-ligand (CD40L) or lipopolysaccharide (LPS). This method significantly improved the generation of Melan-A/MART-1:27-35 and Melan-A/MART-1:26-35(27L) peptide/tumor-specific cells as well as lower frequency tyrosinase:368-376(370D) specific T cells from the PBMC of melanoma patients. T-cell enhancement from activated PBMC cultures was found to be reproducible within individual patients and was observed after the addition of either CD40L or LPS to PBMC cultures. Additionally, PBMC activation improved the detection of tumor-specific precursors from melanoma patients previously immunized with peptides derived from Melan A/MART-1, tyrosinase and gp100. Collectively, these findings describe a novel approach for evaluating patients receiving the cancer vaccines and may provide a useful method for the characterization of novel tumor-associated antigens. PMID- 11730854 TI - mRNA-based electrotransfection of human dendritic cells and induction of cytotoxic T lymphocyte responses against the telomerase catalytic subunit (hTERT). AB - Dendritic cells (DCs) are recognised as the most potent antigen-presenting cells for induction of cellular immune responses, and vaccination with DCs pulsed with antigens has emerged as a promising strategy for generating protective immunity in mammals. We have developed a transfection method that uses in vitro synthesised mRNA and square-wave electroporation for transient expression in DCs and other cell types. The method is highly efficient and produces almost complete transfection of cells in culture. When using mRNA encoding the enhanced green fluorescence protein (EGFP), highest expression in DCs occurred on the second day after transfection and produced a 76-fold increase in mean fluorescence above background. High levels of expression were maintained for at least 5 days post transfection. In comparison, square-wave electroporation of DCs with EGFP plasmid DNA yielded 15% transfected cells and a 28-fold increase of mean fluorescence. DCs transfected with mRNA encoding the telomerase catalytic subunit (hTERT) acquired strong telomerase activity and were capable of eliciting a hTERT specific cytotoxic T lymphocyte (CTL) response in vitro. PMID- 11730855 TI - Engineered expression of the Coxsackie B and adenovirus receptor (CAR) in human dendritic cells enhances recombinant adenovirus-mediated gene transfer. AB - Dendritic cells (DCs) are key antigen-presenting cells (APCs) that act as central modulators of cellular immune responses. Genetic modification of DCs has considerable therapeutic potential in the treatment of a wide spectrum of diseases, including cancer and persistent viral infection. In this report, we show that pre-treatment of DCs with a recombinant adenovirus encoding the major adenovirus receptor, Coxsackie B and adenovirus receptor (CAR), significantly increased the uptake of recombinant adenoviruses (Ads) by primary immature monocyte-derived DCs. This could be correlated with CAR mRNA and surface protein expression. Transduction of DCs by recombinant adenoviruses did not significantly alter cellular viability. Therefore, we propose that pre-treatment of DCs with Ad5-CAR is one strategy to increase the susceptibility of DCs to transduction by recombinant Ads. PMID- 11730856 TI - Direct kinetic assay of interactions between small peptides and immobilized antibodies using a surface plasmon resonance biosensor. AB - A surface plasmon resonance (SPR) protocol is described for the direct kinetic analysis of small antigenic peptides interacting with immobilized monoclonal antibodies (mAb). High peptide concentrations (up to 2.5 microM) and medium mAb surface densities (about 1.5 ng/mm(2)) are needed to ensure measurable binding levels, and fast buffer flow rates (60 microl/min) are required to minimize diffusion-controlled kinetics. Good reproducibility levels in the kinetic constants are obtained under these analysis conditions (standard deviations below 10% of the mean values). Application of this protocol to determine the antigenic ranking of viral peptides shows an excellent agreement between SPR and previous competition enzyme-linked immunosorbent assays (ELISA) on the same peptide/antibody systems. PMID- 11730857 TI - Modelling the national scrapie eradication programme in the UK. AB - In accordance with a policy to eliminate all transmissible spongiform encephalopathies from the food chain, a national untargeted ram breeding programme to eliminate scrapie in the UK is in the final stages of planning. Here we formulate a model of flock-to-flock scrapie transmission, in order to consider the effect of a targeted breeding programme which is in the early stages of consideration. We estimate the size of the susceptible flock population, and discuss implications for potential control programmes. Targeting all rams and ewes in highly susceptible flocks rather than rams in all flocks will eradicate scrapie more quickly, and so is likely to be beneficial as long as suitable penalties or incentives are available to facilitate their identification. A more restricted programme aimed only at highly affected flocks would be much easier to implement and crucially will eradicate scrapie just as quickly. This will leave behind a residue population of susceptible sheep, which could then be gradually removed by a more general breeding programme. PMID- 11730858 TI - The structural identifiability and parameter estimation of a multispecies model for the transmission of mastitis in dairy cows. AB - A structural identifiability analysis is performed on a mathematical model for the coupled transmission of two classes of pathogen. The pathogens, classified as major and minor, are aetiological agents of mastitis in dairy cows that interact directly and via the immunological reaction in their hosts. Parameter estimates are available from experimental data for all but four of the parameters in the model. Data from a longitudinal study of infection are used to estimate these unknown parameters. A novel approach and application of structural identifiability analysis is combined in this paper with the estimation of cross protection parameters using epidemiological data. PMID- 11730859 TI - Success and failure of biphasic shocks: results of bidomain simulations. AB - The mechanisms behind the superiority of optimal biphasic defibrillation shocks over monophasic are not fully understood. This simulation study examines how the shock polarity and second-phase magnitude of biphasic shocks influence the virtual electrode polarization (VEP) pattern, and thus the outcome of the shock in a bidomain model representation of ventricular myocardium. A single spiral wave is initiated in a two-dimensional sheet of myocardium that measures 2 x 2 cm(2). The model incorporates non-uniform fiber curvature, membrane kinetics suitable for high strength shocks, and electroporation. Line electrodes deliver a spatially uniform extracellular field. The shocks are biphasic, each phase lasting 10 ms. Two different polarities of biphasic shocks are examined as the first-phase configuration is held constant and the second-phase magnitude is varied between 1 and 10 V/cm. The results show that for each polarity, varying the second-phase magnitude reverses the VEP induced by the first phase in an asymmetric fashion. Further, the size of the post-shock excitable gap is dependent upon the second-phase magnitude and is a factor in determining the success or failure of the shock. The maximum size of a post-shock excitable gap that results in defibrillation success depends on the polarity of the shock, indicating that the refractoriness of the tissue surrounding the gap also contributes to the outcome of the shock. PMID- 11730860 TI - The effects of diseases on competing species. AB - In this paper we study the dynamics of two competing species when one of them is subject to a disease. In order to keep the model simple, we present it under the strong assumption that the disease cannot cross the species barrier. We answer several important questions about the long term behavior of the environment. In this situation, no population is necessarily wiped out. Also, not every individual of the infected species is bound to contract the disease. Finally, disease-induced persistent cycles for the populations are admissible. PMID- 11730861 TI - 3Beta-brassinosteroid dehydrogenase activity in Arabidopsis and tomato. AB - Enzymatic conversion of 24-epiteasterone to 3-dehydro-24-epiteasterone, representing a reversible step in the biosynthesis of 24R-methyl brassinosteroids, was monitored in vitro in Arabidopsis thaliana and Lycopersicon esculentum using fluorescent tagging and HPLC analysis. In both species 24 epiteasterone was metabolized by the cytosolic fraction of induced root-callus suspension cultures in a NAD dependent manner. This 3beta-dehydrogenation was only slightly inhibited by 12-fold excess of other 3beta-hydroxy intermediates of the early (cathasterone) and late C-6 oxidation pathway (6-deoxo-cathasterone, 6 deoxo-teasterone), indicating a rather specific protein. 3-Dehydro-6 deoxoteasterone and 3alpha-hydroxy intermediates of brassinosteroid biosynthesis did not reduce conversion of 24-epiteasterone to 3-dehydro-24-epiteasterone. In contrast to light-grown cultures the reaction was clearly inhibited in the dark. PMID- 11730862 TI - Partial purification and characterization of a soybean beta-glucosidase with high specific activity towards isoflavone conjugates. AB - A beta-glucosidase with high specific activity towards isoflavone conjugates was purified from soybean [Glycine max] roots by high salt extraction from a low speed centrifugal pellet and subsequent anion and cation exchange chromatography. Purification required stabilization throughout fractionation in 10% glycerol. The enzyme is most likely a dimer (approximate M(r) 165 kDa) with potential subunits of M(r) 80 and/or 75 kDa. The pH and temperature optima are pH 6 and 30 degrees C, respectively. The enzyme was highly heat-stable. Of the various potential effectors examined, silver and mercury ions were the most inhibitory. The IC(50) of silver ions was increased from 140 microM to 14 mM in the presence of 250 microM beta-mercaptoethanol. Glucono-delta-lactone was not strongly inhibitory (IC(50) 24 mM). The activity was highly active against isoflavone conjugates, with a specificity constant 160-1000 fold higher for isoflavone conjugates over the generic chromogenic substrate, p-nitrophenyl beta-glucoside. The enzyme was inactive against the flavonol glycosides tested. The partially purified enzyme had similar K(m) and k(cat) towards 7-O-glucosyl- and 7-O-glucosyl-6"-malonyl isoflavones, suggesting that it may be able to cleave the esterified glucosyl conjugate. We hypothesize that the enzyme is involved in the release of daidzein and genistein, both of which play central roles in soybean defense. PMID- 11730863 TI - Phosphoenolpyruvate carboxylase from cherimoya fruit: properties, kinetics and effects of high CO(2). AB - Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) regulatory properties were studied in non-photosynthetic (mesocarp) and photosynthetic (peel) tissues from cherimoya (Annona cherimola Mill.) fruit stored in air, in order to gain a better understanding of in vivo enzyme regulation. Analyses were also performed with fruit treated with 20% CO(2)-20% O(2) to define the role of PEPC as part of an adaptive mechanism to high external carbon dioxide levels. The results revealed that the special kinetic characteristics of the enzyme from mesocarp--high V(max) and low sensibility to L-malate inhibition - are related to the active acid metabolism of these fruits and point to a high rate of reassimilation of respired CO(2) into keto-acids. With respect to fruit stored in air, PEPC in crude extracts from CO(2)-treated cherimoyas gave a similar V(max) (1.12+/-0.03 microkat x mg(-1) protein), a lower apparent K(m) (68+/-9 microM for PEP) and a higher I(50) of L-malate (5.95+/-0.3 mM). These kinetic values showed the increase in the affinity of this enzyme toward one of its substrate, PEP, by elevated external CO(2) concentrations. The lower K(m) value and lower sensitivity to L-malate are consistent with higher in vivo carboxylation reaction efficiency in CO(2)-treated cherimoyas, while pointing to an additional enzyme regulation system via CO(2). PMID- 11730864 TI - Isolation and properties of arginase from a shade plant, ginseng (Panax ginseng C.A. Meyer) roots. AB - Arginase (EC 3.5.3.1) was purified to homogeneity from root tissues of three-year old ginseng (Panax ginseng C.A. Meyer), shade plant, and was found to be an extraordinarily large molecule relatively stable to heat. The enzyme was decameric having a molecular mass of 352,000 Da, with an optimal temperature and pH of 60 degrees C and 9.5, respectively. Analogues of arginine could not replace it as substrate, and a cysteine residue is at or near the active site. Maximum activity was obtained with Mn(2+) and Co(2+) also activated the proteins, whereas, both agmatine and 5'-deoxy-methylthioadenosine were inhibitors. Specific activities of the enzyme in sliced ginseng roots were increased by plant hormones such as GA(3), IAA, kinetin and putrescine, whereas the activities of the purified enzyme were unaffected by putrescine. Increases in arginase activities by these plant hormones could affect metabolism of polyamine intracellularly. PMID- 11730865 TI - Failed expression of an endo-beta-1,4-glucanhydrolase (cellulase) in a non abscinding mutant of Lupinus angustifolius cv Danja. AB - Cellulase expressions in a normal shedding wild-type and a non-abscinding single gene mutant of Lupinus angustifolius have been studied during ethylene treatments of leaf abscission zone explants. Of the range of different glycohydrolases investigated only the abscission cell-specific beta-1,4-glucanhydrolase (cellulase) was not produced in the non-abscinding mutant. An endo polygalacturonase was induced equally in both wild-type and mutant and other glycohydrolases were equally up-regulated. The abscission cell-specific cellulase induced at shedding of wild-type is antigenically similar to the Phaseolus vulgaris induced leaf abscission pI 9.5 cellulase but with a higher molecular mass (50 kD compared with 48 kD) and like the bean abscission-specific cellulase that of lupin is not glycosylated. Causes of the loss of function of cellulase expression in the non-shedding mutant are discussed. PMID- 11730866 TI - Chemical syntheses of caffeoyl and 5-OH coniferyl aldehydes and alcohols and determination of lignin O-methyltransferase activities in dicot and monocot species. AB - To investigate the substrate preferences of O-methyltransferases in the monolignol biosynthetic pathways, caffeoyl and 5-hydroxy coniferyl aldehydes were synthesized by a new procedure involving a Wittig reaction with the corresponding hydroxybenzaldehydes. The same procedure can also be used to synthesize caffeoyl and 5-hydroxyconiferyl alcohols. Relative O-methyltransferase activities against these substrates were determined using crude extracts and recombinant caffeic acid O-methyltransferase from alfalfa (Medicago sativa), and crude extracts from the model legume Medicago truncatula, tobacco, wheat and tall fescue. Extracts from all these species catalyzed methylation of the various monolignol aldehydes and alcohols more effectively than the corresponding hydroxycinnamic acids. PMID- 11730867 TI - Bioactivity of 25-hydroxy-, 26-hydroxy, 25,26-dihydroxy- and 25,26 epoxybrassinolide. AB - The bioactivity of 25-hydroxybrassinolide, (25S)- and (25R)-26 hydroxybrassinolide, (25S)- and (25R)-25,26-dihydroxybrassinolide, and of (25R) 25,26-epoxybrassinolide was tested in the rice leaf lamina inclination assay. The 25- and (25S)-26-hydroxy derivatives are known metabolites of the naturally occurring phytohormone brassinolide, whereas the other compounds are novel, but closely related, congeners. When tested alone, all showed either no activity or only weak activity at relatively high doses. When coapplied with indole-3-acetic acid (IAA), an auxin that synergizes the effects of brassinosteroids, enhanced bioactivity was observed for each compound. However, even when applied together with IAA, none of the compounds proved more bioactive than brassinolide with or without IAA. We conclude from these results that enzymatic hydroxylation of endogenous brassinolide at C-25 and/or C-26 does not enhance brassinosteroid activity, and so does not comprise an activation pathway in brassinolide biosynthesis. Instead, these hydroxylations result in modest to appreciable metabolic deactivation. PMID- 11730868 TI - Biosynthesis of a hopane triterpene and three diterpenes in the liverwort Fossombronia alaskana. AB - The biosynthesis of the triterpene 22-(30)-hopene-29-acid and the diterpenes 7,17 sacculatadiene-11,12-dial (sacculatal), trans-phytol and a new neoverrucosane type diterpenoid (5-oxo-neoverrucos-(13)-ene) was studied by incorporation of [1 13C]-labelled glucose into axenic cultures of the artic liverwort Fossombronia alaskana. Quantitative 13C NMR spectroscopic analysis of the resulting labelling patterns showed that the isoprene units of the triterpene are derived from the mevalonic acid pathway, whereas the isoprene units of the diterpenes are built up via the methylerythritol phosphate pathway. PMID- 11730869 TI - Trichloroethylene oxidative metabolism in plants: the trichloroethanol pathway. AB - Trichloroethylene (TCE) is a widespread and persistent environmental contaminant. Recently, plants, poplar trees in particular, have been investigated as a tool to remove TCE from soil and groundwater. The metabolism of TCE in plants is being investigated for two reasons: one, plant uptake and metabolism represent an important aspect of the environmental fate of the contaminant; two, metabolism pattern and metabolite identification will help assess the applicability of phytoremediation. It was previously shown that TCE metabolites in plants are similar to ones that result from cytochrome P450-mediated oxidation in mammals: trichloroethanol, trichloroacetate and dichloroacetate. Our measurements indicate that one of these metabolites, trichloroethanol, is further glycosylated in tobacco and poplar. The glycoside was detected in all tissues (roots, stems and leaves) in comparable levels, and was at least 10 fold more abundant than free trichloroethanol. The glycoside in tobacco was identified as the ss-D-glucoside of trichloroethanol by comparison of the mass spectra and the chromatographic retention time of its acetylation product to that of the synthesized standard. Trichloroethanol and its glucoside did not persist in plant tissue once plants are removed from TCE contaminated water, indicating further metabolism. PMID- 11730870 TI - Response of prostaglandin content in the red alga Gracilaria verrucosa to season and solar irradiance. AB - The influence of solar irradiance and seasons on prostaglandin (PG) and arachidonic acid (AA) content in the marine red alga Gracilaria verrucosa (Huds.) Papenf. (unattached form) was investigated. PGA(2), PGE(2), PGF(2), and 15-keto PGE(2) were isolated from the alga, quantitatively analyzed as 4-methyl-7 methoxycoumarin esters by high-performance liquid chromatography, and their chemical structures were confirmed by 1H NMR. In June-September, the PG content in the alga was relatively stable (420 microg/g of dry wt. of PGE(2)+PGF(2); 40 microg/g of PGA(2)) and it increased 1.5 times in October. The highest level of PGs was detected in November (2500 microg/g of PGE(2)+PGF(2); 74 microg/g of PGA(2)) when water temperature was fairly low (5-10 degrees C). Algae grown for five months at 50% of incident photosynthetic active radiation (PAR(0)) contained two times less PGE(2) and PGF(2) than algae grown under natural conditions, but the amount of these PG in algae grown at 5% of PAR(0) was close to the normal level. On the contrary, when algae were grown at 5% of PAR(0) the content of PGA(2) increased up to 4 times compared to algae cultivated at 100% PAR(0). In June-November, the amount of AA in total algal lipids slightly varied from 48.9 to 56.7% and did not virtually depend on the light intensity. The probable reasons of the PG content variation in response to environmental factors are discussed. PMID- 11730871 TI - Potential allelochemicals from Sambucus nigra. AB - Twenty-four aromatic metabolites belonging to cyanogenins, lignans, flavonoids, and phenolic glycosides were obtained from Sambucus nigra. Structures were determined on the basis of their spectroscopic features. Two compounds have been isolated and identified as (2S)-2-O-beta-D-glucopyranosyl-2-hydroxyphenylacetic acid and benzyl 2-O-beta-D-glucopyranosyl-2,6-dihydroxybenzoate. All the compounds have been assayed on dicotyledons Lactuca sativa (lettuce) and Raphanus sativus (radish) and monocotyledon Allium cepa (onion) to test their stimulatory or inhibitory effects on seed germination and radicle elongation. Cyanogenins have a mainly inhibiting effect while lignans stimulate the growth. Some compounds show different effects on dicotyledons and monocotyledons. PMID- 11730872 TI - Alkamides from Artemisia dracunculus. AB - From the aerial parts of Artemisia dracunculus, one known alkamide, pellitorine, two new alkamides neopellitorine A and neopellitorine B, and one known coumarin herniarine were isolated. Structures were elucidated by means of UV, IR, MS, 1H and 13C NMR. These compounds showed insecticidal activity against Sitophilus oryzae and Rhyzopertha dominica at 200 microg/ml concentrations. PMID- 11730873 TI - Scyptolin A and B, cyclic depsipeptides from axenic cultures of Scytonema hofmanni PCC 7110. AB - Two novel cyclic depsipeptides were isolated from axenic cultures of the terrestrial cyanobacterium Scytonema hofmanni PCC 7110 and designated scyptolin A and B. Amino acid analyses in context with mass and 1H/13C NMR spectroscopies revealed a composition typical for heterologous cyanopeptolins but containing the uncommon residue 3'-chloro-N-methyl-Tyr (cmTyr) and a unique sidechain. Scyptolin A and B both consist of the N-acylated peptide But(1)-Ala(2)-Thr(3)-Thr(4)-Leu(5) Ahp(6) (3-amino-6-hydroxy-2-oxo-1-piperidine)-Thr(7)-cmTyr(8)-Val(9), which forms a 19-membered ring by esterification of the carboxyl of Val(9) with the hydroxyl of Thr(4). In scyptolin B, the hydroxyl of the Thr(3) residue is additionally esterified with N-butyroyl-Ala. Both scyptolin A and B exhibit selective inhibition of porcine pancreatic elastase in vitro with IC(50) values of 3.1 microg/ml. PMID- 11730874 TI - Antioxidant capacity of a 3-deoxyanthocyanidin from soybean. AB - Soybean cotyledons directly exposed to UV-C (190-280 nm) contained a colored pigment in those areas of the epidermis directly exposed to UV-C. Ethanolic extracts from UV-C irradiated cotyledons showed a significant peak at 532 nm at pH=10, but not seen at pH=6, successive changes in pH were accompanied by reversible changes in the spectra. The identity of the pigment isolated from soybean cotyledons was established as apigeninidin by comparing the features of standard of a apigeninidin (from sorghum) previously characterized by FAB-MS, UV, HPLC, 1H NMR, and IR spectroscopy. To characterize antioxidant activity of this compound, its ability to scavenge radical species in vitro was tested. In the concentration range tested (up to 200 microg ml (-1)), apigeninidin did not show any scavenger activity towards hydroxyl radical, quinones or NO. However, ascorbyl radical and lipid radicals were effectively quenched in a dose-dependent manner. Overall, UV-C radiation triggers molecular signals that lead in soybean cotyledons to the synthesis and accumulation of an antioxidant pigment, apigeninidin, that shows scavenger activity against ascorbyl and lipid radicals in in vitro studies. PMID- 11730875 TI - Taraxastane-type triterpenoids from Saussurea petrovii. AB - Two taraxastane triterpenoids, i.e. taraxast-20-ene-3beta,30-diol (1) and 20alpha,21alpha-epoxy-taraxastane-3beta,22alpha-diol (2), as well as four known triterpenes taraxast-20(30)ene-3beta,21alpha-diol (3), taraxast-20(30)-ene-3beta ol (4), taraxast-20-ene-3beta-ol (5) and taraxastane-3beta,20alpha-diol (6) were isolated from the Chinese medicinal plant Saussurea petrovii. Their structures were elucidated by spectroscopic methods. These compounds, especially 1 and 2, exhibit significant antitumor and antibacterial activity. PMID- 11730876 TI - Cytotoxic isoflavones from Erythrina indica. AB - Bioassay-directed fractionation of the CH(2)Cl(2)-MeOH (1:1) extract of the stem bark of Erythrina indica, has resulted in the isolation of two new isoflavone derivatives named indicanines D and E together with 11 known compounds including: six isoflavones (genistein, wighteone, alpinumisoflavone, dimethylalpinumisoflavone, 8-prenyl erythrinin C, and erysenegalensein E), one cinnamate (erythrinassinate B), two pentacyclic triterpenes (oleanolic acid and erythrodiol), and two phytosterols (stigmasterol and its 3-O-beta-D glucopyranoside). The structures of the new compounds were elucidated by means of spectroscopic analysis. The in vitro cytocidal activity against KB cells of some of the isolated compounds is also reported. PMID- 11730877 TI - A bis-sesquiterpene and sesquiterpenolides from Inula macrophylla. AB - Macrophyllidimer C, a novel bis-sesquiterpene, in which the two sesquiterpene units are directly connected by a C-C bond, and eight other sesquiterpenolides were obtained from the bark of Inula macrophylla. Seven of these, macrophyllilactones A-G, are new eudesmanolide- and elemanolide-type sesquiterpenes. Their structures were determined on the basis of spectroscopic evidence and chemical reaction. PMID- 11730878 TI - Dibenzofurans from the cultured lichen mycobionts of Lecanora cinereocarnea. AB - From the cultures of the spore-derived mycobionts of the lichen Lecanora cinereocarnea, five dibenzofurans, 3,7-dihydroxy-1,9-dimethyldibenzofuran, 2 chloro-3,7-dihydroxy-1,9-dimethyldibenzofuran, 2,8-dichloro-3,7-dihydroxy-1,9 dimethyldibenzofuran, 3-hydroxy-7-methoxy-1,9-dimethyldibenzofuran, and 2-chloro 7-hydroxy-3-methoxy-1,9-dimethyldibenzofuran, were isolated. Their structures were determined by spectroscopic methods. PMID- 11730879 TI - Diterpenoids from Euphorbia paralias. AB - Two segatane diterpenoids containing a bicyclic [4,3,1] ring system were isolated, together with 11 known diterpenoids, four segetanes, five jatrophanes and two paralianes, from the aerial parts of Euphorbia paralias. The structures of the new compounds were established by spectroscopic means, including by 2D NMR and CD spectroscopic analyses. The antifeedant and antiviral activities of the isolated compounds are also described. PMID- 11730880 TI - Antifungal highly oxygenated guaianolides and other constituents from Ajania fruticulosa. AB - Three highly oxygenated guaianolides were isolated from the aerial parts of Ajania fruticulosa along with 17 known phytochemicals including a triterpene (alpha-amyrin), two plant sterols (beta-sitosterol, daucosterol), four flavonoids (axillarin, centaureidin, santin and 5,7,4'-trihydroxy-3,3'-dimethoxyflavone), and ten sesquiterpenes [1alpha-hydroperoxy-4beta,8alpha,10alpha,13 tetrahydroxyguaia-2-en-12,6alpha-olide, 1alpha-hydroperoxy-4alpha,10alpha dihydroxyguaia-9alpha-angeloyloxyguaia-2,11(13)-dien-12,6alpha-olide, 3beta,4alpha-dihydroxyguaia-11(13),10(14)-dien-12,6alpha-olide, 1alpha,4alpha,10alpha-trihydroxy-9alpha-angeloyloxyguaia-2,11(13)-dien-12,6alpha olide, 1beta,2beta-epoxy-3beta,4alpha,10alpha-trihydroxy-guaia-11(13)-en 12,6alpha-olide and 2-oxo-8alpha-hydroxyguaia-1(10),3,11(13)-trien-12,6alpha olide, ketoplenolide B, alantolactone, 9beta-hydroxyeudesma-4,11(13)-dien-12-oic acid and 9beta-acetoxyeudesma-4,11(13)-dien-12-oic acid]. The structures of the three guaianolides were elucidated by a combination of spectroscopic methods (EIMS, HREIMS, COSY, HMQC, HMBC and NOESY) as 1beta,2beta-epoxy 3beta,4alpha,8beta,10alpha-tetrahydroxyguaia-11(13)-en-12,6alpha-olide (1), 1beta,2beta-epoxy-3beta,4alpha,9alpha,10alpha-tetrahydroxyguaia-11(13)-en 12,6alpha-olide (2) and 1beta,2beta-epoxy-10alpha-hydroperoxy-3beta,4alpha,8beta trihydroxyguaia-11(13)-en-12,6alpha-olide (3), respectively. Antifungal bioassay of all isolates showed that guaianolides 1, 2, 3, and 1beta,2beta-epoxy 3beta,4alpha,10alpha-trihydroxyguaia-11(13)-en-12,6alpha-olide were inhibitory to the growth of Candida albicans with MICs being 20, 20, 20, and 40 microg/ml, respectively. PMID- 11730884 TI - Doses to the embryo and fetus from intakes of radionuclides by the mother. A report of The International Commission on Radiological Protection. AB - In its Publications 56, 67, 69, 71, and 72, ICRP has provided age-specific biokinetic models and, using those models, compiled dose coefficients (doses per unit intake) for intakes of radionuclides by members of the public. Committed effective doses for the inhalation or ingestion of radionuclides by workers who are occupationally exposed were given in Publication 68 based on the most recent dosimetric and biokinetic models for adults. Additionally, a compilation of dose coefficients for both workers and members of the public has been issued on a CD ROM. The present report complements that series by addressing doses to the embryo/fetus after intakes of radionuclides by a female member of the public or a female worker, before or during pregnancy. Ingestion and inhalation of selected radionuclides of the 31 elements for which age-dependent biokinetic models were provided in the previous reports are considered. These elements are H, C, S, Ca, Fe, Co, Ni, Zn, Se, Sr, Zr, Nb, Mo. Tc, Ru, Ag, Sb, Te, I, Cs, Ba, Ce, Pb, Po, Ra. Th, U, Np, Pu, Am, and Cm. New biokinetic and dosimetric models for calculating doses to the developing embryo and fetus are developed and used in conjunction with the models for infants, children, and adults presented in the previous reports. The models used take account of transfer of radionuclides across the placenta, distribution and retention of radionuclides in fetal tissues, growth of the fetus, and photon irradiation from radionuclides in the placenta and maternal tissues. Human and animal data are used as available in the development of these models. Intake scenarios comprising single or continuous maternal intakes are taken into account in the compilation of effective dose coefficients following ingestion or inhalation of the radionuclides considered. A CD-ROM with more comprehensive information on doses from inhalation of different particle sizes, tissue doses, and doses at various times after birth has been developed concurrently with the report and will be available shortly. The report does not consider doses to the offspring due to intakes of radionuclides in maternal milk and external irradiation from the mother's body after birth. The radiation sensitivity of the offspring is not discussed. PMID- 11730893 TI - Prion diseases: copper deficiency states associated with impaired nitrogen monoxide or carbon monoxide transduction and translocation. AB - Literature concerning prion diseases and Cu metabolism was examined to determine merits of various suggestions concerning the relationship between these diseases and altered Cu metabolism. There are a number of recent suggestions that the normal non-pathogenic form of the prion protein (PrP(C)) contains Cu while the abnormal pathogenic form of this protein, PrP(SC), lacks Cu. Results of experiments showing oxidant sensitivity in the presence of ionically bonded Cu and millimolar concentrations of hydrogen peroxide were found to lack relevance. Demonstrating superoxide disproportionation and a correlation with cellular Cu2Zn2SOD activity is relevant and consistent with a role for PrP(C) in Cu endocytosis. There are also a number of recent suggestions that PrP(C) has a role in nerve transmission. Serum from mice that lack cellular PrP(C) was found to have an elevated Cu content consistent with a response to overcome an inflammatory disease. Attempts to induce a 'transmissible' form of prion disease requiring intracerebral injections of somewhat purified brain homogenates were found lacking in support for an etiology occurring as the result of oral ingestion of supposedly 'infected' tissues. It is suggested that PrP(C) is a normal Cu-dependent cuproglycoprotein of unknown function that may have a role in facilitating normal nitrogen monoxide- or carbon monoxide-mediated biochemistry. PMID- 11730894 TI - Copper-nuclease efficiency correlates with cytotoxicity for the 4-methoxypyrrolic natural products. AB - The DNA-targeting activities of the 4-methoxypyrrolic natural products, that include prodigiosin (1), tambjamine E (2), and the blue pigment (3), have been compared using fluorescence spectroscopy to study DNA binding and agarose gel electrophoresis to assess their ability to facilitate oxidative copper-promoted DNA cleavage. Fluorescence emission titration of 3 with calf-thymus DNA (CT-DNA) shows that the natural product occupies a site size (n) of ca. two base pairs and possesses an affinity constant (K) of approximately 6x10(5) x M(-1). Similar to prodigiosin (1), the blue pigment 3 was found to facilitate oxidative double strand DNA (dsDNA) cleavage without the aid of an external reducing agent. Quantitation of ds- (n2) and ss- (n1) breaks provided n1:n2 ratios of approximately 8-12, which were significantly greater than the number expected from the accumulation of ss-breaks (approximately 120). This was contrasted by the nicking activity of tambjamine E (2), which only generates ss-breaks in the presence of copper. The superior copper-nuclease activity of 1 and 3 also correlated with their superior anticancer properties against leukemia (HL-60) cells. These results are discussed with respect to the mode of cytotoxicity by the 4-methoxypyrrolic natural products. PMID- 11730895 TI - New methyl pyruvate thiosemicarbazones and their copper and zinc complexes: synthesis, characterization, X-ray structures and biological activity. AB - By reacting thiosemicarbazides substituted on the aminic nitrogen with both alkyl or aryl groups, and methyl pyruvate a new group of methylpyruvate thiosemicarbazones (Hmpt) derivatives was obtained. These ligands were then treated with copper and zinc inorganic salts. All isolated compounds were characterized using spectroscopic methods. The single crystal structural analysis of the ligands Me-Hmpt x 0.5H2O 1, Et-Hmpt x H2O 2, Ph-Hmpt 5, Meph-Hmpt 6 showed that only compound 6 presents significant deviation from planarity. The X-ray structure of [Zn(Me-Hmpt)Cl2] x H2O 8 showed that in this complex Me-Hmpt behaves as a neutral ligand SNO terdentate and that the penta coordination is achieved by chloride ions according to spectroscopic and elemental analyses. On the basis of the analytical data the same behavior is proposed for the other zinc complexes. All the ligands in copper complexes seem to be monodeprotonated; nevertheless the same SNO behavior is expected. Tests on cell proliferation of human leukemic cell line U937 showed that the copper complex Cu(Et-mpt)Cl x H2O is the most active compound among those reported even though it is not able to induce apoptosis. PMID- 11730896 TI - Cobalt(II) and copper(II) binding of Bacillus cereus trinuclear phospholipase C: a novel 1H NMR spectrum of a 'Tri-Cu(II)' center in protein. AB - The phosphatidylcholine-preferring phospholipase C from Bacillus cereus (PC PLC(Bc)) is a tri-Zn enzyme with two 'tight binding' and one 'loose binding' sites. The Zn2+ ions can be replaced with Co2+ and Cu2+ to afford metal substituted derivatives. Two Cu2+-substituted derivatives are detected by means of 1H NMR spectroscopy, a 'transient' derivative and a 'stable' derivative. The detection of sharp hyperfine-shifted 1H NMR signals in the 'transient' derivative indicates the formation of a magnetically coupled di-Cu2+ center, which concludes that the Zn2+ ions in the dinuclear (Zn1 and Zn3) sites are more easily replaced by Cu2+ than that in the Zn2 site. This might possibly be the case for Co2+ binding. Complete replacement of the three Zn2+ ions can be achieved by extensive dialysis of the enzyme against excess Cu2+ to yield the final 'stable' derivative. This derivative has been determined to have five-coordinated His residues and an overall S'=1/2 spin state with NMR and EPR, consistent with the formation of a tri-Cu2+ center (i.e. a di-Cu2+/mono-Cu2+ center) in this enzyme. The binding of substrate to the inert tri-Cu2+ center to form an enzyme-substrate (ES) complex is clearly seen in the 1H NMR spectrum, which is not obtainable in the case of the native enzyme. The change in the spectral features indicates that the substrate binds directly to the trinuclear metal center. The studies reported here suggest that 1H NMR spectroscopy can be a valuable tool for the characterization of di- and multi-nuclear metalloproteins using the 'NMR friendly' magnetically coupled Cu2+ as a probe. PMID- 11730897 TI - Synthesis, characterization and antitumor activity of quinolone-platinum(II) conjugates. AB - A new series of quinolone-platinum(II) conjugates, [Pt(Q'-NH2)(dmso)X2] and cis [Pt(Q"-en)X2], where Q' and Q" are quinolones (flumequine, nalidixic acid or oxolinic acid) linked to monodentate and bidentate amine ligands, respectively, and X2 is Cl2 or 1,1-cyclobutanedicarboxylate, have been synthesized with the aim of examining the synergetic antitumor activity of quinolone intercalation and platinum(II) chelation. The complexes were characterized by elemental analyses and IR and multinuclear (1H and 195Pt) NMR spectroscopies, and then subjected to in vitro and in vivo bioassays using the leukemia L1210 cell line. PMID- 11730898 TI - Mechanisms of nitric oxide release from nitrovasodilators in aqueous solution: reaction of the nitroprusside ion ([Fe(CN)5NO]2-) with L-ascorbic acid. AB - Nitric Oxide (NO) is a very diverse endogenous molecule and is responsible for a number of mammalian physiological activities. Sodium nitroprusside (SNP) is a long known hypotensive agent due to its ability to produce NO in situ. In this study, we have carried out a detailed investigation on the kinetics and mechanism of the decomposition of SNP by L-ascorbic acid to afford NO in aqueous medium. Spectrophotometric and electrochemical methods were employed in these studies. Experiments were performed under an argon atmosphere and reaction rates were strongly pH dependent and increases with pH. NO release increases up to about pH 7 after which it decreases at higher pHs. The two ionized forms of ascorbate present in aqueous solution reduce SNP in the order A2->>HA->>H2A (H2A=L-ascorbic acid) to release NO. The outer-sphere reduction of SNP by ascorbate involved three clear stages with NO being released in the last stage. Our kinetic results also show catalysis by group 1 alkali metal ions to increase down the group from Li+ to Cs+. A detail study presented here, strongly illustrate the possibility of the biological antioxidant, L-ascorbic acid, to play a vital role in the in situ metabolism of nitrosocompounds such as SNP to produce NO. PMID- 11730899 TI - Two-dimensional electrophoretic analysis of sperm antigens recognized by sperm immobilizing antibodies detected in infertile women. AB - In this study, high resolution two-dimensional (2-D) gel electrophoresis was used to identify human sperm antigens recognized by the sera from infertile women having sperm immobilizing (SI) antibodies. Two-D gel electrophoresis was employed to separate Percoll purified human sperm proteins using isoelectric focusing (IEF), followed by polyacrylamide gel electrophoresis (PAGE). Sperm proteins were transferred to the nitrocellulose membranes and immunoblotted with seven sera from infertile women with high titers of SI antibodies and 6 sera from those without SI antibodies. The blots were compared to the 2-D composite image of human sperm proteins [Sperm Protein Encyclopedia] and sperm surface index and the sperm surface proteins recognized by infertile sera were identified. Fifty-two human sperm surface proteins reacted with sera containing SI antibodies, while 35 of these were reactive with the SI-negative control sera. The average numbers of protein spots reacted with test and control sera were 24.6 and 15.0 respectively. A subset of sperm surface proteins which were unique to the SI antibodies were identified by the following criteria; the sperm protein spots which were highly reactive with the infertile sera containing SI antibodies but not reactive with any of the SI-negative infertile sera. The coordinates of 4 prominent immunoreactive sperm proteins were considered as possibly relevant to antibody mediated female infertility. PMID- 11730900 TI - Molecular and immunological characteristics of sperm antigens involved in egg binding. AB - Development of a vaccine(s) based on sperm antigens represents a promising approach to contraception. The utility of an antigen in immunocontraception is contingent upon sperm/testis-specificity and its involvement in the fertilization process. Since, the sperm-zona pellucida (ZP) recognition and binding constitutes the most important event in the fertilization process, molecules involved at this site are attractive candidates for immunocontraception. Using hybridoma technology, subtractive hybridization, and differential display technology, our laboratory has delineated several sperm antigens. These antigens have testis specific expression and have a role in the fertilization process. The cDNAs encoding for the antigens have been cloned and sequenced. Among these, the fertilization antigen (FA-1) is particularly interesting, because it is involved in immuno-infertility in humans. Using the phage peptide display technique, a novel dodecamer sequence of a approximately 72+/-5 kD antigen, designated as YLP(12), that is testis-specific and involved in human sperm-ZP recognition/binding, was identified. A synthetic 12-mer peptide was generated based on this sequence. In the hemizona assay, YLP(12) peptide and its monovalent Fab' antibodies specifically and significantly inhibited human sperm-ZP binding. Furthermore, the presence of specific antibodies reactive with YLP(12) peptide, were identified in the serum and seminal plasma of immuno-infertile men. Thus, FA 1 and YLP(12) are promising target antigens for the development of contraceptive vaccines as well as for specific diagnosis and treatment of male infertility. PMID- 11730901 TI - Transcriptional regulation of spermiogenesis: insights from the study of the gene encoding the acrosomal protein SP-10. AB - Spermiogenesis is the terminal differentiation process of the male germ cell during which haploid spermatids acquire unique structures such as the acrosome and flagellum and undergo extensive cellular reorganization. Although well described morphologically, the molecular mechanisms underlying spermiogenesis are not well understood. The SP-10 gene, which codes for the acrosomal protein SP-10, has been well characterized in mice and men. This single copy gene is localized to syntenic regions of chromosomes 9 and 11 in mouse and human, respectively. The SP-10 gene is testis-specific, and is transcribed and translated in round spermatids. The differentiation marker SP-10 serves as a useful model to address questions regarding the regulation of round spermatid-specific gene transcription and acrosome biogenesis. This paper defines the temporal pattern of SP-10 gene expression during spermiogenesis and reviews the work done on analysis of the SP 10 promoter. Transgenic mice demonstrated that either the -408/+28 or the 266/+28 region of the SP-10 promoter could drive round spermatid-specific expression of a GFP reporter gene whereas the -91/+28 region lacked promoter activity. The transgene expression mimicked the spatial and temporal patterns of expression of the endogenous SP-10 gene. Surprisingly, none of the transgenic lines showed expression of GFP in tissues other than testis. Given the complexity of eukaryotic transcriptional regulation, the fact that a short 294-bp promoter is capable of conferring developmental stage- and cell type-specific transcription of a gene is intriguing and paradoxical. PMID- 11730902 TI - Molecular analysis of the Y chromosome AZFc region in Japanese infertile males with spermatogenic defects. AB - Cytogenetic and molecular studies of azoospermic and oligozoospermic males have suggested the presence of azoospermia factors (AZF) in the human Y chromosome. Deletion in three Y chromosomal regions--AZFa, AZFb and AZFc--has been reported to disrupt spermatogenesis and cause infertility. Several candidate genes responsible for spermatogenesis have been identified in these regions and some of them are thought to be functional in human spermatogenesis. Here we report on clinical and molecular studies of Y chromosome micro-deletions in Japanese. In these studies the data from 157 infertile Japanese men with azoospermia and oligozoospermia was analyzed and divided into 5 categories based on spermatozoa count. Sixteen sets of primers were used for polymerase chain reaction (PCR) to amplify sequence tagged site markers. One common deletion in the AZFc region was identified in infertile men. On the other hand, no deletions around the AZFc region were identified in fertile men. Japanese infertile men in our study had a common deletion in the AZFc region of the Y chromosome. A genomic clone was obtained by PCR screening of the P1 phage artificial chromosome (PAC) library. This clone was analyzed by Southern blotting using a PCR amplified probe of sY240. Our analysis of the genomic sequence of the clone suggests that this locus may contain specific genes for spermatogenesis. PMID- 11730903 TI - Reduction in the level of hyaluronan binding protein 1 (HABP1) is associated with loss of sperm motility. AB - Hyaluronan binding protein 1 (HABP1) was reported to be present on human sperm surface and its involvement in fertilization has already been elucidated (Mol. Repro. Dev. 38 (1994) 69). In the present communication, we report a significant reduction in the level of this protein in sperms from asthenozoospermic and oligozoospermic patients as compared to normozoospermic one. Further evidence of the absence of HABP1 in sperms, having motility <20% is documented, which again is a determining factor for fertilization. HABP1 was quantitatively determined using anti-HABP1 antibody from sperm extracts isolated from semen samples of both the fertile and infertile groups demonstrating low sperm motility. Sperm samples with low motility revealed a significant reduction in the level of HABP1 in immunoblot detection as well as immunolocalization experiment. It suggests that decreased HABP1 level may be associated with low motility of sperms, which in turn might cause infertility in the patient. Thus, the sperm surface HABP1 level can be correlated with the degree of sperm motility, an important criteria for fertilization. PMID- 11730904 TI - Oolemmal proteomics. AB - Fertilization is defined as a series of gametic interactions in which capacitated sperm must first penetrate the egg vestments and then bind to and fuse with the egg plasma membrane (oolemma). The molecular basis of sperm-egg binding and fusion has yet to be elucidated due, in part, to how little is known about the array of proteins residing on the oolemma. Proteomics is an emerging area of research that directly evaluates protein expression by resolving, identifying, quantitating, and characterizing proteins utilizing a variety of techniques including high resolution two-dimensional polyacrylamide gel electrophoresis (2D PAGE), tandem mass spectrometry, and computer analysis. Our research group has utilized 2D PAGE to begin building a mouse oocyte proteomic database, with over 500 silver-stained proteins being resolved and digitized to date. Cell-surface labeling with biotin has identified a subset of 80 putative egg surface proteins. Amino acid microsequences from over 30 of the surface-labeled proteins has been obtained by tandem mass spectrometry. Sequences from eight of these proteins do not match any sequences from protein and DNA databanks, indicating that these proteins are novel. Our major current research goal is to clone, characterize, and express the novel proteins that are shown to be ovary-specific and investigate their functional roles in sperm-egg interaction. PMID- 11730905 TI - Primate recombinant zona pellucida proteins expressed in Escherichia coli bind to spermatozoa. AB - To delineate the role of individual zona pellucida (ZP) glycoproteins during sperm-oocyte interaction, bonnet monkey (bm; Macaca radiata) ZPA (bmZPA), ZPB (bmZPB), and ZPC (bmZPC) have been cloned without native signal sequence and transmembrane-like domain, and expressed in Escherichia coli. Recombinant proteins have been purified from the inclusion bodies in presence of low concentration of chaotropic agent (2 M urea) and high pH (pH 12), and subsequently refolded in presence of oxidized and reduced glutathione. Binding of the recombinant refolded zona proteins to bonnet monkey spermatozoa in an indirect immunofluorescence assay revealed that recombinant bmZPC binds to the head region of the capacitated spermatozoa but does not bind to the acrosome reacted spermatozoa. Recombinant bmZPB binds to the principal segment of the acrosomal cap of capacitated bonnet monkey spermatozoa. After induction of acrosome reaction by calcium ionophore A23187, the binding of recombinant bmZPB shifts to the equator, post-acrosome and midpiece of the spermatozoa. bmZPA binds to the principal segment of capacitated spermatozoa but the binding shifts to the equatorial segment, tip of the inner acrosomal membrane and midpiece in acrosome reacted spermatozoa. These studies suggest that polypeptide backbone is sufficient for the binding of ZPA, ZPB and ZPC to spermatozoa in non-human primates. Further studies with recombinant glycosylated zona proteins will help in delineating the role of carbohydrate moieties for higher affinity binding of the ligand to spermatozoa and subsequent signal transduction pathways. PMID- 11730906 TI - Involvement of carbohydrate molecules on zona pellucida in human fertilization. AB - Although the involvement of several receptors and ligand molecules in sperm-zona interaction in many species have been proposed, there has been a little analysis of the kinetics between these molecules during the interaction. In the present study, we applied the detection method using surface plasmon resonance (SPR) by a BIAcore apparatus for the analysis of the putative receptor-ligand interaction of sperm-egg binding. Mannose-BSA or [man](5)-[GlcNAc](2)-Asp was immobilized on the surface of a sensor chip. When concanavalin A (Con A) was delivered to each of two different sensor chips to evaluate their usefulness, the resonance signal after sample injection onto a [man](5)-[GlcNAc](2)-Asp-fixed chip decreased rapidly than the mannose-BSA-fixed chip. However, the amount of binding for Con A during the injection onto the [man](5)-[GlcNAc](2)-Asp-fixed chip was high. When acid sperm extracts (acid extracts) and fractions through a CM column, containing protease activity (protease fractions), were delivered to the mannose-BSA-fixed chip, the SPR signal during the injection was not obviously changed compared with that of the control. However, when sperm samples were delivered to the [man](5) [GlcNAc](2)-Asp-fixed chip, the SPR response during the injection was enormous. These results suggest that the [man](5)-[GlcNAc](2)-Asp-fixed chip is more useful than the mannose-BSA-fixed chip for investigating the interactions with sperm extracts and that the sensitive method using SPR by a BIAcore apparatus is applicable for the analysis of the putative receptor-ligand interaction of sperm egg binding. PMID- 11730907 TI - Contraceptive potential of synthetic peptides of zona pellucida protein (ZPA). AB - Previously, we produced a fertilization-blocking monoclonal antibody (MAb-5H4) to find a candidate peptide for a contraceptive vaccine. MAb-5H4 recognized a linear amino acid sequence of ZPA (No. 50-67) in pigs, humans and rabbits. In the present study, 18mer peptides corresponding to the sequence were conjugated with diphtheria toxoid as a carrier protein before immunization in rabbits. All three antisera recognized human zona pellucida on testing by immunofluorescent staining method. The two produced against human and rabbit peptides effectively inhibited human sperm binding to the zona pellucida, but the antiserum against the pig peptide did not. The former two peptides include an identical sequence (LDPEKLTL) of the minimum binding motif for MAb-5H4, but the latter peptide includes one amino acid replacement (K to N) in the sequence. It is thus concluded that a synthetic peptide including the sequence of LDPEKLTL could be a feasible candidate for developing a contraceptive vaccine for humans. PMID- 11730908 TI - The contraceptive potential of ZP3 and ZP3 peptides in a primate model. AB - It has been known for some time that antibodies raised against ZP3, the major component of the glycoprotein shell that surrounds all mammalian oocytes, can successfully inhibit sperm-egg interaction in vitro. In our own studies using the non-human primate Callithrix jacchus, active immunisation was successfully achieved when homologous or heterologous ZP3 was used as an immunogen. However this long-term suppression of fertility was at the expense of ovarian function. An ovarian pathology was observed which was characterised by a disruption of folliculogenesis and depletion of the primordial follicle pool. Adverse auto immune reactions have also been observed in mice following induction of immunity to mouse ZP3. Following careful selection of B-cell epitopes on mouse ZP3, peptide vaccines were formulated which could circumvent these adverse side effects and induce reversible infertility in actively immunised mice. To identify similar epitopes on primate ZP3, epitope mapping studies were performed and several candidate regions of the molecule were identified. These were incorporated into chimeric peptide vaccines and administered as single or triple peptide vaccines. Active immunisation successfully induced antibodies that bound exclusively to the zona pellucida of marmoset and human ovarian sections. These antibodies were able to suppress human sperm-egg binding by up to 60% in vitro. Encouragingly, no adverse side effects on ovarian function were observed following long-term immunisation however, no loss of fertility was consistently observed in vivo. Thus considerable research is still required to identify a combination of ZP epitopes that will induce reversible infertility in the absence of any ovarian dysfunction. PMID- 11730909 TI - Role of seminal plasma phospholipid-binding proteins in sperm membrane lipid modification that occurs during capacitation. AB - Bovine seminal vesicles secrete a family of similar proteins designated BSP-A1, BSP-A2, BSP-A3 and BSP-30-kDa (collectively called bovine seminal plasma (BSP) proteins). The biochemical properties of these proteins are well documented and considerable progress has been made concerning their biological role. At ejaculation these BSP proteins bind to the sperm surface. The binding sites on the sperm surface have been identified as choline phospholipids (specifically phosphatidylcholine (PC), phophatidylcholine plasmalogen (PC plasm) and sphingomyelin (SPM)) composed of sperm plasma membrane. Our previous studies have shown that the BSP proteins interact specifically with heparin and high-density lipoproteins (HDL), the capacitation factors in bovine. In addition, we have shown that the BSP proteins potentiate epididymal sperm capacitation induced by heparin and HDL. Recently, we showed that the BSP proteins stimulated cholesterol and phospholipid efflux from the sperm membrane. Furthermore, the lipid efflux from sperm is dependent on BSP protein concentration and duration of incubation. The loss of membrane cholesterol is an important step in the capacitation process. These results together indicate that BSP proteins play an important role in sperm membrane lipid modification events that occur during sperm capacitation. PMID- 11730910 TI - Signal transduction pathways in human spermatozoa. AB - Following ejaculation, sperm functions are regulated by interactions with the environment found in the female genital tract. Here spermatozoa become 'capacitated' (i.e. they acquire the capability of completing the acrosome reaction and successfully fertilizing the egg), through a series of surface and intracellular transformations occurring during a process known as capacitation. The limited number of spermatozoa that eventually reach the oocyte must then cross the surrounding cumulus cells before contacting the zona pellucida, undergoing the acrosome reaction and finally fertilizing the oocyte. All the interactions between sperm and their diverse environments are mediated by intracellular signaling pathways. Virtually all the signaling systems known to operate in somatic cells, with the exception of those implying nuclear activity, have been found in spermatozoa, although in some cases the function(s) of the pathway has not been clearly proved. PMID- 11730911 TI - Novel signaling pathways involved in sperm acquisition of fertilizing capacity. AB - Capacitation is a complex series of molecular events that occurs in sperm after epididymal maturation and confers on sperm the ability to fertilize an egg. This process can be mimicked in vitro in defined media, the composition of which is based on the electrolyte concentration of oviductal fluid. In most cases, capacitation media contain energy substrates, such as pyruvate, lactate and glucose, a cholesterol acceptor (usually serum albumin), NaHCO(3), Ca(2+), low K(+), and physiological Na(+) concentrations. The mechanism of action by which these compounds promote capacitation is poorly understood at the molecular level; however, some molecular events significant to the initiation of capacitation have been identified. For example, capacitation correlates with cholesterol efflux from the sperm plasma membrane, increased membrane fluidity, modulations in intracellular ion concentrations, hyperpolarization of the sperm plasma membrane and increased protein tyrosine phosphorylation. These molecular events are required for the subsequent induction of hyperactivation and the acrosome reaction. This review discusses the recent progress that has been made in elucidating mechanisms which regulate sperm capacitation. PMID- 11730912 TI - Role and regulation of intracellular calcium in acrosomal exocytosis. AB - Calcium influx is required for the mammalian sperm acrosome reaction, an exocytotic event occurring in the sperm head after binding to the egg. Prior to this binding, the spermatozoon undergo, in the female reproductive tract, a series of biochemical transformations, collectively called capacitation. The first event in capacitation is the elevation of intracellular calcium, bicarbonate and hydrogen peroxide, which collectively activate adenylyl cyclase to produce cyclic-AMP, which activates protein kinase A. During capacitation, there is an increase in the membrane-bound phospholipase C, and this binding is highly stimulated by adding epidermal growth factor to the cells. We suggest that zona-pellucida binds to at least two different receptors in the sperm head plasma membrane. One is a G(i)-coupled receptor that can activate phospholipase Cbeta(1) and might regulate adenylyl cyclase to further enhance cyclic-AMP levels. The cyclic AMP activates protein kinase A to open a calcium channel in the outer acrosomal membrane, resulting in a relatively small rise in cytosolic calcium. This rise in Ca(2+) leads to activation of phospholipase Cgamma, which is coupled to the second tyrosine kinase receptor. The products of phospholipase C activity, diacylglycerol and inositol-trisphosphate (IP(3)), will lead to activation of protein kinase C (PKC) and IP(3)-receptor. PKC will open a calcium channel in the plasma membrane and IP(3) will activate the calcium channel in the outer acrosomal membrane, leading to a higher increase in cytosolic calcium. In addition, the depletion of calcium in the acrosome will activate a store-operated Ca(2+) channel, resulting in a very fast increase in cytosolic calcium (300-500 nM), leading to membrane fusion and completing the acrosome reaction. PMID- 11730913 TI - Signaling through protein kinases during egg activation. AB - Following penetration of the zona pellucida, the mammalian spermatozoon binds and fuses with the egg plasma membrane, thereby fertilizing the egg that is still arrested at the second metaphase. Fertilization initiates in the egg a sequence of events referred to as 'egg activation'. An initial increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) appears to be the very early cellular event observed which leads to the cortical granules exocytosis and resumption of meiosis. Various treatments can induce parthenogenetic activation mimicking at least part of the fertilization events. Similar to somatic cells, studies in mammalian eggs suggest that signal transduction pathways mediate egg activation. The initial increase in [Ca(2+)](i) appears to be critical for egg activation. However, other messengers such as protein kinase C (PKC) and protein tyrosine kinases (PTKs), were suggested as possible inducers of some aspects of egg activation. In the present work, studies concerning the involvement of protein kinases during egg activation in our laboratory and in others are summarized. PMID- 11730914 TI - Oocyte-specific genes regulate follicle formation, fertility and early mouse development. AB - Gene products expressed in oocytes play important roles in folliculogenesis, fertilization and pre-implantation development. Factor in the Germline, alpha (FIGalpha) is a basic helix-loop-helix (bHLH) transcription factor first detected in oocytes at E13.5 that persists in adults. Female mice lacking FIGalpha are unable to form primordial follicles which results in massive depletion of oocytes and sterility. FIGalpha is also required for expression of the zona pellucida genes that encode ZP1, ZP2, and ZP3. Mice lacking ZP1 form structurally abnormal zonae and have decreased fecundity. Mice lacking ZP3 have no zona matrix despite the presence of the other two zona proteins. Although, folliculogenesis occurs in Zp3 null mice, few eggs are ovulated and females are sterile. Transgenic mice expressing human ZP3 have been crossed with Zp3 null mice and reconstitute a chimeric zona pellucida matrix (moZP1, moZP2, huZP3). Unexpectedly, human sperm do not bind to 'humanized' zonae pellucidae in vitro, but mouse sperm do. Although, late in oogenesis, oocytes becomes transcriptionally inactive and maternal RNA is degraded, the activation of early development requires pre existing maternal products from the egg. Maternal antigen that embryos require (Mater) is a single-copy gene that is transcribed in growing oocytes and, although its transcripts are degraded during meiotic maturation, MATER protein persists into the blastocyst. Female mice lacking this 125 kDa cytoplasmic protein produce no off-spring because of an embryonic block at the early cleavage stage. Thus, Mater represents one of the few documented maternal effect genes in mammalian development. PMID- 11730915 TI - Interactions between zona pellucida glycoproteins and sperm proacrosin/acrosin during fertilization. AB - Fertilization is one of the most specific and carefully regulated cell-cell interactions in the animal body and is determined to a large extent by compatibility between ligand and receptor molecules on the surface of each gamete. On the zona pellucida (ZP), sperm receptor activity is associated with glycoproteins ZP3 (primary receptor for acrosome-intact sperm) and ZP2 (secondary receptor for acrosome-reacted sperm) but their complementary binding proteins on sperm are less well defined. In this communication we review the evidence for proacrosin as a secondary ZP binding protein. Proacrosin/acrosin binds non enzymically to ZP glycoproteins. Binding is a strong ionic interaction between polysulphate groups on ZP glycoproteins (probably on their carbohydrate moieties) and basic residues on the surface of proacrosin. The stereochemistry of the reactants is crucial and determines to a large extent the affinity of binding. Site-directed mutagenesis and a 3D-structural analysis of boar and ram acrosin have identified 2 clusters of basic residues potentially involved in binding. A polysulphonated anticancer drug, suramin, has been shown to bind strongly to proacrosin/acrosin and to inhibit sperm-egg binding in vitro. In the mouse model, 125I-ZP2 and 3H-suramin bind approximately 65% less effectively to acrosin 'null' sperm than to wild-type sperm. Neither ZP2 nor suramin bind to acrosome intact sperm and can, therefore, only exert their effects after exposure of the acrosomal contents. Overall, this combination of biochemical, genetic and functional data supports the hypothesis that proacrosin is a multifunctional protein with a significant role in retaining acrosome-reacted sperm on the ZP surface long enough to enable ZP penetration to begin. PMID- 11730916 TI - Gene expression profiling of human GV oocytes: an analysis of a profile obtained by Serial Analysis of Gene Expression (SAGE). AB - A gene expression profile of the human GV oocyte has recently been established by Serial Analysis of Gene Expression (SAGE). A significant number of the genes identified in this profile had not previously been associated with mammalian oocytes. We sought to confirm gene matches by RT-PCR amplification of candidate transcripts using mouse eggs. Attention focused on receptors, proteins involved in apoptosis, and cytoskeletal proteins. Two receptors found in the human catalogue, CCR6 and PAR3, were not found in mouse eggs, whereas myosin light chain, LLGL, beta-actin, 5HT receptor, bad, bak, DFF45, and Caspase homologue (cash) were. Individual SAGEtags can match more than one gene and, in some cases, more than ten. Examination of transcript sequences that generate multiple gene assignments identified a common denominator of short interspersed elements or Alu sequences. For reasons which are, as yet, unclear, the human GV oocyte SAGE catalogue contains relatively high abundances of SAGEtags in Alu sequences. This may reflect normal expression of Alu-containing genes in eggs or upregulated expression of Alu elements following stress. The degeneracy of gene matches in SAGE generated by Alu sequences makes independent confirmation of candidate genes essential. PMID- 11730918 TI - Newly identified endometrial genes of importance for implantation. AB - The mammalian uterus is normally not receptive to embryo implantation except during the very limited 'window of implantation'. To identify genes that may be responsible for this phenomenon the technique of RNA differential display (DD PCR) was applied to implantation and inter-implantation sites on day 4.5 of pregnancy in the mouse, the time at which the blastocyst becomes attached to the endometrium. Three of these genes were identified as splicing factor SC35, calbindin-D9k and monoclonal non-specific suppressor factor beta (MNSFbeta). Expression of SC35 mRNA, which is responsible for removal of introns from pre mRNA, is much higher in implantation than in interimplantation sites during pregnancy. Expression of alternatively spliced mRNAs for SC35 is differentially regulated by early pregnancy and steroid hormones. By contrast, calbindin-D9k, a regulator of calcium, is upregulated by progesterone and its mRNA increases in the uterus during early pregnancy compared with during the cycle, although it is significantly lower in implantation sites than in interimplantation sites on days 4.5-5.5 of pregnancy, but subsequently becomes barely detectable in both sites. The mRNA for calbindin-D9k is predominantly in endometrial luminal epithelium. MNSFbeta, a cytokine involved in regulation of the immune system, showed lower expression at implantation sites than interimplantation sites on day 4.5 of pregnancy, when embryos first attach to the uterus and initiate implantation, and on day 5.5 when implantation has advanced. Immunohistochemically, the protein was localized to endometrial stromal cells in the non-pregnant uterus, but disappeared as decidualization progressed. The precise roles of these three proteins in the process of embryo implantation remains to be determined. Homologues of the proteins may contribute to the development of the 'window of implantation' in the human and hence be appropriate targets for new post-coital contraceptives or may be manipulated to improve fertility. PMID- 11730917 TI - Regulation of peri-attachment embryo development in the golden hamster: role of growth factors. AB - The molecular regulation of mammalian peri-implantation development is complex and difficult to study in vivo. We successfully cultured hamster blastocysts through hatching and peri-attachment stages, using a chemically defined medium, HECM-2h. Using this system, we showed that a species-specific, embryonic cysteine like protease is involved in blastocyst hatching and that the process is modulated by growth factors. In particular, heparin binding-epidermal growth factor (HB-EGF) or leukemia inhibitory factor (LIF) enhance blastocyst hatching, and the former also improves attachment and trophoblast outgrowth. We observed interesting changing patterns of expression of mRNA and/or immunoreactive protein for EGF, HB-EGF, LIF and transforming growth factor-beta (TGF-beta) in the embryo and/or endometrial tissue, during peri-implantation development. Together, it appears that hamster blastocyst hatching, attachment and trophoblast outgrowth are regulated by autocrine and/or paracrine growth factors, produced by the embryo-endometrial tissues. PMID- 11730919 TI - Blastocyst-endometrium interaction at implantation in the rhesus monkey. PMID- 11730920 TI - A brief review of recent data on some cytokine expressions at the materno-foetal interface which might challenge the classical Th1/Th2 dichotomy. AB - Focussing attention on cytokines at the materno-foetal interface represents one of the major advances made in the field. This owes much to the visionary views of Tom Wegmann, and to the changes brought about in the field by immunotrophism and Th1/Th2 paradigms. We review these briefly and also point out some emerging problems.However, a certain number of newly discovered cytokines do not fit into the classical Th1/Th2 dichotomy. Yet, by their capacity to activate or downregulate NK cells, by their action on adhesion molecules, and by their regulatory effects on the vascularisation process, they are of possible interest within the materno-foetal relationship. Therefore, as a first step, we have undertaken a systematic study of the expression of IL-11, IL-12, IL-13, IL-15, IL 16, IL-17 and IL-18 in the uterus, the peri-implantation embryo, and later on decidual and placental tissues throughout pregnancy. These cytokines were detected in every case, with, in each case, a precise localisation, which will be detailed, and which indeed suggests important regulatory functions, especially during implantation. In some cases, as will be shown in the peri-implantation uterus, those cells are perfectly expressed by uterine GMG-NK-like cells. Comparative ELISAs and quantitative RT-PCR have been or are being conducted, but already the expression patterns that are observed, and the very precise window of appearance that is observed for some of the GMG NK-like cells, either around or in the implanting embryo, as well as the complexity of the respective distributions, strongly suggest that, as useful as it certainly was for a while, the Th1/Th2 paradigm must now be considered as an over-simplification. Rather, the existing data point to sequential windows and are suggestive of a system where an extreme complexity is allied to very precise timing and tuning. They also suggest that the materno-foetal relationship is not simply maternal tolerance of a foreign tissue, but a series of intricate mutual cytokine interactions governing selective immune regulation and also control of the adhesion and vascularisation processes during this dialogue. PMID- 11730921 TI - Network of cytokines, integrins and hormones in human trophoblast cells. AB - Trophoblast cells of the developing embryo are unique not only for transporting oxygen and nutrients from the mother to the fetus but also for their array of other functions throughout the pregnancy beginning from the attachment of the blastocyst to the endometrium during the process of implantation, its well regulated invasion in the uterine tissue, proliferation, differentiation and immuno-endocrine functions which subsequently maintain the pregnancy. Using human trophoblast cells in culture, we have tried to understand the molecular mechanisms which regulate such a variety of functions for trophoblast cells. Our RT-PCR studies show that trophoblast cells express the laminin and collagen receptors: integrins alpha1 and alpha2, which are both stimulated by IL-1 and IL 6. These two cytokines, also synthesised by the trophoblast cells themselves, act in an autocrine/paracrine manner to induce their own expression. In addition, IL 1 expression seems to be modulated by a large variety of cytokines and growth factors usually present in the uterine milieu whereas IL-6 expression appears to be significantly stimulated by growth factors like EGF and bFGF only. Hormones like estrogen, progesterone and hCG exhibit a general negative modulation in the expression of IL-1 and IL-6. Since, both IL-1 and IL-6 are known to be involved in the proliferation, invasion and differentiation of trophoblast cells, they might be the key factors involved in trophoblast functions. PMID- 11730922 TI - Autoantibodies to contact proteins in patients with recurrent pregnancy losses. AB - The kallikrein-kinin system or plasma contact system consists of three essential plasma proteins. These are coagulation factor XII, prekallikrein and high molecular weight kininogen. Deficiencies of these proteins are not associated with clinical bleeding despite marked prolongation of in vitro surface-activated coagulation time. Paradoxically, studies suggest that contact proteins have anticoagulant, profibrinolytic functions in a physiologic millieu, on endothelial cells. Recently, evidence has accumulated for the presence of the kallikrein kinin system or plasma contact system in the fetoplacental unit. Kinins which are released within the placenta may play a role in regulating placental blood flow. This suggests that the plasma contact system may also have an important role in pregnancy. Several studies have reported the presence of autoantibodies to the contact proteins in patients with recurrent early pregnancy losses. Disruption of this system may be a risk factor for early gestational losses. PMID- 11730923 TI - Interest in preeclampsia for researchers in reproduction. AB - Gestational hypertension/preeclampsia, is a major disease of human reproduction, with 10% of human births being affected. It is due to the failure of extravillous cytotrophoblast to invade the maternal uterine spiral arteries to a sufficient depth at the second physiological invasion around the 14-16th week of gestation, inducing poor vascular exchanges between the mother and the placenta. The rise of blood pressure in the human mother is then a compensatory mechanism to increase the exchanges and try to save the fetus from poor supplies. Indeed, it is only in the late 1970s that a puzzling phenomenon has been described: in human pregnancy, in contrast with other mammals, implantation of the embryo occurs by two physiological invasions of the cytotrophoblast inside the uterine wall: (a) at the beginning of pregnancy after fecundation (like all mammals); (b) then follows an apparent long pause (6-8 weeks) and (c) late at the end of first trimester (14 16th week) of gestation, a second very deep (1/3 of the uterine wall) invasion. This two-wave physiological endovascular trophoblast invasion represents a remarkable immunological placental-maternal interaction. Moreover, preeclampsia which has been considered as 'the disease of primigravidae' during all the XXth century may be in fact associated with new paternity especially in couples conceiving very shortly after the beginning of their sexual relationship. 'Primipaternity', rather than primigravidity, is probably the leading cause of preeclampsia. Comprehension in the near future of the physiological immunological tolerance in normal pregnancies and immunological rejection (preeclampsia) of the second trophoblastic invasion will give the biological clue of this puzzling disease, real plague of human reproduction. PMID- 11730924 TI - Human malignant cell death by apoptosis-inducing nucleosides from the decidua derived CD57(+)HLA-DR(bright) natural suppressor cell line. AB - The CD57(+)HLA-DR(bright) natural suppressor (57.DR-NS) cell line derived from human decidual tissue mediated apoptosis of human leukemia Molt4 and carcinoma BeWo/GCIY cells but not human fibroblast WI-38 cells, and apoptosis-inducing nucleosides (AINs) appeared to be involved. Six AINs were released into 57.DR-NS cell culture media and were isolated by the combination of physicochemical procedures of C18 preparative column, thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC). Subsequently, we demonstrated that AINs could induce apoptosis in the human malignant Molt4/BeWo/GCIY cell line but not human normal WI-38 fibroblasts. Apoptosis was characterized by DNA strand breaks and activation of the caspase cascade, especially caspase-3. The administration of AINs into GCIY tumor bearing SCID mice culminated in suppression of tumor growth due to apoptosis of tumor cells. PMID- 11730925 TI - Immunopathology of Bartonella vinsonii (berkhoffii) in experimentally infected dogs. AB - Following natural infection with Bartonella, dogs and humans develop comparable disease manifestations including endocarditis, peliosis hepatis, and granulomatous disease. As the immunologic response to infection in these hosts has not been clearly established, data presented here was derived from the experimental infection of six specific pathogen free (SPF) beagles with a known pathogenic strain of Bartonella. Six dogs were inoculated intravenously with 10(9)cfu of B. vinsonii ssp. berkhoffii and six control dogs were injected intravenously with an equivalent volume of sterile saline. Despite production of substantial levels of specific antibody, blood culture and molecular analyses indicated that Bartonella established chronic infection in these dogs. Flow cytometric analysis of monocytes indicated impaired bacterial phagocytosis during chronic Bartonella infection. There was also a sustained decrease in the percentage of CD8+ lymphocytes in the peripheral blood. Moreover, modulation of adhesion molecule expression (downregulation of L-selectin, VLA-4, and LFA-1) on CD8+ lymphocytes suggested quantitative and qualitative impairment of this cell subset in Bartonella-infected dogs. When compared with control dogs, flow cytometric analysis of lymph node (LN) cells from B. vinsonii infected dogs revealed an expanded population of CD4+ T cells with an apparent naive phenotype (CD45RA+/CD62L+/CD49D(dim)). However, fewer B cells from infected dogs expressed cell-surface MHC II, implicating impaired antigen presentation to helper T cells within LN. Taken together, results from this study indicate that B. vinsonii establishes chronic infection in dogs which may result in immune suppression characterized by defects in monocytic phagocytosis, an impaired subset of CD8+ T lymphocytes, and impaired antigen presentation within LN. PMID- 11730926 TI - Production and characterisation of monoclonal antibodies specific for chicken interleukin-2. AB - Using genetic immunisation of mice, we produced antibodies against chicken interleukin-2 (ChIL-2), the first produced against a non-mammalian interleukin. After a final injection with a recombinant ChIL-2 protein, two stable hybridoma cell lines were established which secreted monoclonal antibodies (MAbs) against this cytokine. Specific binding of the two MAbs to recombinant ChIL-2 produced by Escherichia coli and COS-7 cells was demonstrated in an indirect ELISA, Western blotting and dot blots. Both of them were able to neutralise the biological activity of the ChIL-2, but neither allowed the detection of ChIL-2 by flow cytometry. PMID- 11730927 TI - Detection of cellular cytokine mRNA expression during orf virus infection in sheep: differential interferon-gamma mRNA expression by cells in primary versus reinfection skin lesions. AB - In sheep infected with the parapoxvirus orf virus, primary infection orf skin lesions developed and resolved within 8 weeks. Reinfection lesions were smaller and resolved within 3 weeks. The host response in the skin was characterized by an accumulation of neutrophils, dendritic cells, CD4+ T cells, CD8+ T cells, B cells and T19+ gammadelta T cells. The magnitude of this accumulation paralleled orf virus replication in the skin. In situ hybridization was used to detect cells expressing interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4) mRNAs in orf skin. Cells expressing IL-4 mRNA were not detected at any time after infection. Cells expressing IFN-gamma mRNA were detected after reinfection but not after primary infection. Cells expressing TNF alpha mRNA included epidermal cells, vascular endothelium and uncharacterized cells that increased more rapidly in the skin after reinfection compared to primary infection. The results are consistent with a prominent role for IFN-gamma in the host immune response controlling the severity of the disease. PMID- 11730928 TI - The correlation of virus-specific interferon-gamma production and protection against classical swine fever virus infection. AB - The level of antigen-specific interferon-gamma (IFN-gamma) production can be used as an indicator of cellular immunity. In this study, we investigated the role of cellular immune response in protection against classical swine fever virus (CSFV). Pigs were vaccinated once with CSFV vaccine and challenged 6 days post vaccination (dpv). Vaccinated animals had significantly higher CSFV-specific IFN gamma secreting cells than the unvaccinated pigs (p<0.05) at the time of challenge and were protected against CSFV infection, whereas the control pigs died within 14 days post-infection (dpi). In the second experiment, pigs were vaccinated once with either CSFV vaccine or CSFV vaccine combined with Aujeszky's disease (AD) vaccine and challenged at 140 dpv. All vaccinated pigs developed both CSFV-specific, cellular and antibody responses and were protected against CSFV infection. However, differences in cellular, but not antibody, responses were observed in the two vaccinated groups. The group vaccinated with CSFV vaccine developed a significantly higher number of CSFV-specific, IFN-gamma secreting cells (p<0.05), exhibited a shorter fever period and less pathological changes, when compared with the group vaccinated with the combined vaccine. The kinetics of IFN-gamma production, following challenge in the two vaccinated groups, were also different. Taken together, our results indicated that CSFV specific, IFN-gamma production could be detected early after antigen exposure and correlated with protection against CSFV challenge. Our findings highlight the role of cellular immune responses in porcine anti-viral immunity. PMID- 11730929 TI - Semi-quantitative analysis of multiple cytokines in canine peripheral blood mononuclear cells by [correction of zby] a single tube RT-PCR. AB - Cytokines play an important role in the regulation of the immune system, but low circulating levels in plasma make routine measurement a difficult task. A new methodology based on single tube RT-PCR has been developed to determine the expression of multiple canine cytokines (TNF-alpha, IL-2, IFN-gamma, IL-18, IL-4, IL-6 and IL-10) using primers and protocols designed allow specific amplification of the mRNAs. The technique is performed in one tube in two consecutive steps, a specific transcription of the mRNA of a given cytokine and amplification of the corresponding gene by PCR. The technique was used to analyse the mRNA cytokine profile of peripheral blood mononuclear cells (PBMCs) from healthy dogs using two approaches: (i) analysis of PBMC isolated ex vivo; (ii) analysis of PBMC after in vitro cultures with or without the mitogen ConA. The samples were separated in agarose gels and the intensity of ethidium bromide signals quantified using standard video imaging equipment. Results were interpreted as the ratio of cytokine to GAPDH expression. The results obtained show that the method is easy to use and reproducible. Therefore, this method of monitoring the mRNA cytokine expression might be an useful tool for understanding the immune response in dogs. PMID- 11730930 TI - Elevated extrahepatic expression and secretion of mammary-associated serum amyloid A 3 (M-SAA3) into colostrum. AB - Mammary-associated serum amyloid A 3 (M-SAA3) was secreted at highly elevated levels in bovine, equine and ovine colostrum and found at lower levels in milk 4 days postparturition. N-terminal sequencing of the mature M-SAA3 protein from all the three species revealed a conserved four amino acid motif (TFLK) within the first eight residues. This motif has not been reported to be present in any of the hepatically-produced acute phase SAA (A-SAA) isoforms. Cloning of the bovine M-Saa3 cDNA from mammary gland epithelial cells revealed an open reading frame that encoded a precursor protein of 131 amino acids which included an 18 amino acid signal peptide. The predicted 113 residue mature M-SAA3 protein had a theoretical molecular mass of 12,826Da that corresponded with the observed 12.8kDa molecular mass obtained for M-SAA3 in immunoblot analysis. The high abundance of this extrahepatically produced SAA3 isoform in the colostrum of healthy animals suggests that M-SAA3 may play an important functional role associated with newborn adaptation to extrauterine life and possibly mammary tissue remodeling. PMID- 11730931 TI - Pre-translational regulation of neutrophil L-selectin in glucocorticoid challenged cattle. AB - L-selectin (CD62L) gene expression in neutrophils is commonly referred to as "constitutive" because circulating neutrophils require a constant supply of this adhesion molecule for continuous trafficking into peripheral tissues. Under normal circumstances, marginating blood neutrophils and neutrophils that become activated for migration into infected tissues rapidly shed surface CD62L that is ligated to the vascular endothelium. However, this does not shut down CD62L gene expression because these cells continue to express surface CD62L. In contrast, glucocorticoid challenges resulting from stress and hormone injections result in gradual and chronic down-regulation of CD62L on the surface of blood neutrophils. Rather than being associated with migration, this type of CD62L down-regulation associates with pronounced neutrophilia and increased susceptibility to infections. Nothing is currently known about glucocorticoid regulation of CD62L expression in neutrophils. In other cell systems, however, this steroid hormone binds to cytoplasmic glucocorticoid receptors (GR) that influence expression of glucocorticoid-responsive genes at multiple pre-translational levels. Thus, the hypothesis of the present study was that glucocorticoid challenge suppresses CD62L mRNA expression in blood neutrophils. Suppressed CD62L gene expression might help explain the chronic down-regulation of surface CD62L in neutrophils and accompanying neutrophilia. The main objectives of the study were to monitor neutrophil CD62L mRNA abundance before and during subtle and severe glucocorticoid challenges and to determine if CD62L mRNA expression correlates with degree of glucocorticoid challenge. Parturient dairy cows and dexamethasone treated steers were used as models of subtle and severe (respectively) glucocorticoid challenges. Data presented from both models support the hypothesis and show for the first time that glucocorticoids regulate neutrophil CD62L at a pre-translational level. Results also showed that inhibited CD62L mRNA expression correlated precisely with down-regulated surface expression of CD62L on neutrophils and peak neutrophilia during severe glucocorticoid challenge. Therefore, results of this study indicate that bovine neutrophils are highly sensitive to the blood environment, displaying full capacity to alter CD62L gene expression and trafficking patterns in response to changing glucocorticoid levels. This may serve animals well when heightened inflammatory responses begin to lead to tissue damage, but may be detrimental to overall health if animals are exposed to opportunistic pathogens while stressed or undergoing glucocorticoid therapy. Although this study did not elucidate how glucocorticoids inhibit neutrophil CD62L mRNA expression, presented data implicate GR as possibly being involved because neutrophils from cattle in both models expressed GR mRNA. Further in vitro studies using purified populations of neutrophils will be required to determine if GR is directly involved in glucocorticoid regulation of CD62L gene expression and, if so, at what level. PMID- 11730932 TI - Molecular cloning and expression analysis of feline melanoma antigen (MAGE) obtained from a lymphoma cell line. AB - Melanoma antigens (MAGE) are regarded as inducing tumor-specific immune response and thought to be potential therapeutical agents for cancer immunotherapy. We hereby report the cloning of feline MAGE cDNA obtained from a lymphoma cell line derived from cat malignant lymphoma, and its expression pattern in tumor and normal tissues. The cDNA encoding the MAGE is 1668 base pairs (bp) in length, and contains an open reading frame (ORF) of 936 bp encoding a protein of 311 amino acids. The predicted amino acid sequence has 29-46% of homology with other MAGE proteins from human and mouse. mRNA transcripts for the feline MAGE were detected in certain tumors, but not in adult cat normal tissues except in testis, by reverse transcription polymerase chain reaction (RT-PCR) analysis. This indicates that the expression pattern of feline MAGE mRNA is similar to those of other MAGE family genes in tumors and normal tissues. PMID- 11730933 TI - Characterisation of tryptase and a granzyme H-like chymase isolated from equine mastocytoma tissue. AB - Mast cell proteinases are important inflammatory mediators in man and other species, but until now there has been no investigation of the nature of equine mast cell proteinases. These studies describe the purification and characterisation of two proteolytic components from equine mastocytoma tissue, detected using chromogenic substrates for trypsin and chymotrypsin. Following chromatographic purification, the trypsin-like component was found to be equine mast cell tryptase by N-terminal amino acid sequencing, showing a close similarity with human tryptase-beta (85% identity over 20 residues). It also had similar subunit molecular size (34-36kDa by SDS-PAGE) and substantially similar cleavage specificity to human tryptase-beta with the substrates tested. A 32kDa chymotrypsin-like component was also purified from mastocytoma extract, and termed equine mast cell proteinase-1 (eqMCP-1). The N-terminal amino acid sequence of eqMCP-1 was very similar to human granzyme H (95% over 19 residues). Rabbit antisera directed against tryptase and eqMCP-1 both detected equine mast cells by immunohistochemistry, and will be of use in future clinical studies of the relevance of mast cell proteinases in equine allergic disease. PMID- 11730934 TI - Functional interaction between cyclin T1/cdk9 and Puralpha determines the level of TNFalpha promoter activation by Tat in glial cells. AB - In addition to its stimulatory effect on transcription of the HIV-1 LTR, the early protein of HIV-1, Tat, exhibits detrimental effects on the CNS by deregulating the expression of several cytokines and immunomodulators including TNFalpha. Activation of the viral promoter by Tat requires several cellular proteins including cyclin T1 and its partner, cdk9, which upon association with the TAR sequence of the LTR, forms a complex that enhances the activity of RNA polymerase II. Here, we examined the involvement of cyclin T1/cdk9 in Tat mediated transcriptional activation of the TNFalpha promoter which has no TAR sequence. Results from transfection of human astrocytic cells revealed that both cyclin T1 and cdk9 stimulate the basal promoter activity of TNFalpha, although the level of such activation is decreased in the presence of Tat. Ectopic expression of Puralpha, a brain-derived regulatory protein which binds to Tat, enhanced the basal level of TNFalpha transcription, yet exerted a negative effect on the level of Tat activation of the TNFalpha promoter. The antagonistic effect of Puralpha and Tat upon the TNFalpha promoter was diminished in the presence of cyclin T1 and cdk9, suggesting cooperativity of Puralpha with cyclin T1 and cdk9 in Tat activation of the TNFalpha promoter. Results from protein-protein binding studies showed the interaction of Puralpha with both cyclin T1 and cdk9 through distinct domains of Puralpha which are in juxtaposition with each other. Interestingly, the site for cyclin T1 binding within Puralpha is adjacent to the region which is important for Tat/Puralpha association. In light of these observations, we propose a model which ascribes a bridging role for Puralpha in assembling Tat, cyclin T1, and cdk9 around the promoter region of TAR-negative genes such as TNFalpha, which is responsive to Tat activation. PMID- 11730935 TI - T cell-mediated neuroprotection involves antithrombin activity. AB - Functional loss after injury to the mammalian central nervous system (CNS) has been attributed not only to the immediate loss of neurons but also to secondary neuronal degeneration caused by the toxicity of physiological compounds present in abnormally high amounts as a result of the injury. One such compound appears to be the protease thrombin. Here we show that the beneficial effect of T cells directed against myelin self-antigens can be attributed, at least in part, to the ability of these 'autoimmune' T cells to produce antithrombin III. Using transgenic mice lacking the thrombin receptor PAR-1, we also present molecular evidence indicating that down-regulation of PAR-1 by genetic manipulation leads to increased post-traumatic survival of CNS neurons. We further show that the ability of autoimmune T cells to produce thrombin inhibitors and to exert post traumatic neuroprotection are both independent of their PAR-1 expression. These findings suggest that thrombin plays a key role in post-injury neuronal survival, and that its post-traumatic toxicity can be down-regulated by appropriate alteration of the amounts of PAR-1 receptors or of antithrombin III, the latter achieved by T cell-mediated autoimmunity. PMID- 11730936 TI - The dynamics of bone marrow stromal cells in the proliferation of multipotent hematopoietic progenitors by substance P: an understanding of the effects of a neurotransmitter on the differentiating hematopoietic stem cell. AB - Communication within the hematopoietic-neuroendocrine-immune axis is partly mediated by neurotransmitters (e.g. substance P, SP) and cytokines. SP mediates neuromodulation partly through the stimulation of bone marrow (BM) progenitors. This study shows that SP, through the neurokinin-1 receptor, stimulates the proliferation of primitive hematopoietic progenitors: cobblestone-forming cells (CAFC, CD34+). This effect is optimal when macrophage is included within the fibroblast support. Indirect induction of IL-1 could be important in the proliferation of CAFC colonies by SP. Phenotypic and functional studies suggest that SP might directly interact with the CD34+/CD45(dim) population. These studies indicate that SP can initiate a cascade of biological responses in the BM stroma and stem cells to stimulate hematopoiesis. PMID- 11730937 TI - IL-1 receptor type I gene expression in the amygdala of inflammatory susceptible Lewis and inflammatory resistant Fischer rats. AB - Lewis (LEW/N) and Fischer (F344/N) rats have different responses to inflammatory and behavioral stressors due to differences in hypothalamus-pituitary-adrenal (HPA) axis function. For example, LEW/N rats are more sensitive to restraint, inflammation and experimentally induced autoimmunity due to decreased HPA activity. The HPA axis response to peripheral inflammation is mediated, at least in part, by IL-1beta and its receptor, IL-1 type I (IL-1RI). Here, we studied the distribution of IL-1RI mRNA in the brains of LEW/N and F344/N rats, and demonstrated that IL-1RI mRNA expression has significantly increased in the basolateral nucleus (BLA) of the amygdala of LEW/N rats. These findings suggest that strain-specific HPA axis responses may be mediated by extrahypothalamic pathways. PMID- 11730938 TI - Microarray analysis of gene expression in multiple sclerosis and EAE identifies 5 lipoxygenase as a component of inflammatory lesions. AB - Multiple sclerosis (MS) is a demyelinating disease of the central nervous system characterized by lesions that are areas of blood-brain barrier breakdown, inflammation and myelin damage. To identify genes that contribute to lesion pathology, we have compared gene expression in MS lesions and in brains of mice with experimental allergic encephalomyelitis (EAE) with that in normal white matter. Gene expression was analyzed by cDNA microarrays consisting of 2798 human genes. One of the genes found to be upregulated in both MS lesions and EAE brains was 5-lipoxygenase (5-LO), a key enzyme in the biosynthesis of the proinflammatory leukotrienes. The presence of 5-LO in MS lesions was confirmed by immunohistochemistry and indicated that 5-LO was primarily contained within macrophages. Although these findings are not specific for MS, they identify a potentially important component of pro-inflammatory activity in the demyelinating process in MS and suggest a possible target for anti-inflammatory therapy in MS. PMID- 11730939 TI - Topical application of the cornea post-infection with plasmid DNA encoding interferon-alpha1 but not recombinant interferon-alphaA reduces herpes simplex virus type 1-induced mortality in mice. AB - A study was undertaken to compare the efficacy of recombinant interferon (rIFN) alphaA to plasmid DNA encoding IFN-alpha1 against ocular herpes simplex virus type 1 (HSV-1) infection. The topical application of rIFN-alphaA (100-300 units/eye) onto the cornea of mice subsequently infected 24 h later with HSV-1 antagonized viral-induced mortality. The enhancement in cumulative survival in the rIFN-alphaA-treated mice correlated with a reduction of viral titers recovered in the eye and trigeminal ganglion (TG) at 3 and 6 days post-infection. The protective effect was site-specific such that when rIFN-alphaA was administered orally or intranasally, no efficacy against HSV-1 was observed. However, the protective effect was time-dependent. Specifically, when the rIFN alphaA (100-1000 units/eye) was administered at 24 h post-infection, no protective effect was observed against HSV-1 compared to the vehicle-treated group. In contrast, plasmid DNA (100 microg/eye) containing the IFN-alpha1 transgene showed significant protection when topically applied 24 h post infection. Although the transgene was found to traffic distal from the site of application (eye), including the trigeminal ganglion and the spleen where CD11b(+) and CD11c(+) cells express the transgene, the migration of the transgene did not correlate with efficacy. Collectively, the results suggest that naked DNA encoding type I IFN applied post-infection provides a greater degree of protection against ocular HSV-1 infection in comparison with recombinant protein effectively antagonizing viral replication and spread. PMID- 11730940 TI - Structure of epitopes recognized by the antibodies to alpha(181-192) peptides of neuronal nicotinic acetylcholine receptors: extrapolation to the structure of acetylcholine-binding domain. AB - Using the alpha(181-192) peptides of neuronal nicotinic acetylcholine receptor (nAChR) and Ala-substituted peptide analogues, amino acid residues critical for specific monoclonal antibody (mAb) binding were identified. By means of 2D nuclear magnetic resonance (2D-NMR) analysis followed by molecular modeling, it was found that mAb binding resulted in stabilization of the free alpha3(181-192) peptide flexible conformation yielding an extended structure with residues 6-11 of the peptide being in direct contact with the Ab. Since the Ab binds the native AChR as well, it is suggested that the corresponding fragment of AChR alpha3 subunit is exposed to solution and also appears in extended conformation. PMID- 11730942 TI - Caspase-3 activity is present in cerebrospinal fluid from patients with traumatic brain injury. AB - Loss of neurons after traumatic brain injury (TBI) might involve dysregulated apoptosis. Activation of caspase-3 is one hallmark of apoptosis. Therefore, caspase-3 activity (cleavage of DEVD-afc) was measured in cerebrospinal fluid (CSF) samples (n=113) from 27 patients with TBI at day 1 to 14 after trauma. Caspase-3 activity was detected in 31 (27.4%) CSF samples with highest values (> 5.5 microM/min) seen at day 2-5 after trauma. No caspase-3 activity was found in serum from patients or CSF from controls. The presence of activated caspase-3 in CSF suggests ongoing apoptotic processes during traumatic brain injury. PMID- 11730941 TI - Substance P enhances HIV-1 replication in latently infected human immune cells. AB - Substance P (SP) is a potent modulator of neuroimmunoregulation. SP receptors are present on human monocytes and T lymphocytes, and SP alters the function of these immune cells. We investigated the effects of SP on HIV-1 replication in latently infected human immune cells. SP significantly enhanced HIV-1 replication in the latently infected promonocytic cell line (U1) and T lymphocyte line (ACH-2) stimulated with tumor necrosis factor (TNF-alpha). When added to these cells in combination with TNF-alpha, SP also enhanced HIV-1 gag gene expression in U1 and ACH-2 cells. This stimulatory effect of SP was associated with the activation of HIV-LTR (long terminal repeat) driven chloramphenicol acetyltransferase (CAT) gene expression, and could be blocked by pretreatment of U1 and ACH-2 cells with an SP receptor antagonist RP-67,580, indicating specific SP receptor-mediated regulation. Furthermore, the addition of SP to the cultures of latently infected peripheral blood mononuclear cells isolated from HIV-1-infected patients enhanced HIV-1 gag gene expression. Thus, SP may play a potentially important role as a positive regulator of HIV-1 replication in latently infected monocytes and lymphocytes. These observations may have significant implications toward understanding the role of neuropeptide SP in the immunopathogenesis of HIV-1 infection and AIDS. PMID- 11730943 TI - Anti-GD1a antibodies from an acute motor axonal neuropathy patient selectively bind to motor nerve fiber nodes of Ranvier. AB - Acute motor axonal neuropathy (AMAN) is associated with high titer anti-GD1a antibodies. We have found that very high titer IgG anti-GD1a antibodies (Ab) from one AMAN patient selectively bind to motor, but not sensory, nerve nodes of Ranvier. Binding is abolished by preadsorption with GD1a. Sera negative for Ab do not immunostain motor and sensory nerve roots. We have also found that botulinum toxin A (BTA), which binds to GD1a, stains both motor and sensory nerve nodes of Ranvier. Our results strongly support the pathogenetic role of anti-GD1a antibodies in AMAN. Why BTA also binds to sensory fibers still remains to be elucidated, although the different size of BTA and its specificity to other gangliosides present in sensory axons might represent important factors. PMID- 11730944 TI - Differential immune response to gangliosides in Guillain-Barre syndrome patients from Japan and The Netherlands. AB - Anti-ganglioside antibodies are consistently found in Guillain-Barre syndrome (GBS) patients from different geographical parts of the world. Several studies indicated differences in relative frequencies of anti-ganglioside reactivity and isotype distribution between GBS patients from Asia and from Europe. We investigated antibody reactivity against the gangliosides GM1, GM1b and GalNAc GD1a in GBS patients from Japan and The Netherlands in two different laboratories. The proportion of GBS patients with anti-ganglioside antibodies did not differ between the two countries. GBS patients from The Netherlands more frequently had cross-reacting anti-GalNAc-GD1a/anti-GM1b antibodies and a stronger IgM anti-ganglioside response. Our findings indicate that geographical determined factors, dependent on either the host or the triggering infectious agent, determine the isotype distribution and fine specificity of anti ganglioside antibodies in GBS patients. PMID- 11730945 TI - Single-cell analysis of cytokine production shows different immune profiles in multiple sclerosis patients with active or quiescent disease. AB - Peripheral blood mononuclear cells of multiple sclerosis (MS) patients were stimulated with myelin basic protein (MBP) together with anti-CD28 monoclonal antibody and staphylococcal enterotoxin B to optimize cytokine production by antigen-specific cells. Type 1 (IL-2, IL-12, IFNgamma) and pro-inflammatory (TNFalpha, IL-1beta, IL-6) cytokines were augmented in CD4+, CD8+, and CD14+ cells of acute MS patients and of patients undergoing disease reactivation. These cytokines were reduced in IFNbeta-treated and in stable MS patients; type 2 cytokines (IL-4, IL-10) were increased in these patients. Similar immune profiles are seen in MS patients in whom remission is naturally or pharmacologically (IFNbeta) achieved. Cytokine alterations are particularly evident in CD14+ cells, underlying their critical role in the modulation of the immune response. PMID- 11730946 TI - Glatiramer acetate and IFN-beta act on dendritic cells in multiple sclerosis. AB - Glatiramer acetate (GA; Copolymer 1; Copaxone) and interferon-beta (IFN-beta) modulate the course of multiple sclerosis (MS), but probably by different mechanisms. GA, a mixture of synthetic peptides, is believed to act as an altered peptide ligand with inhibitory effects on autoreactive T cells and promoting Th2 cells. It is unknown whether GA affects dendritic cells (DCs), which, besides strong antigen presenting capacity, orchestrate Th1 and Th2 responses. IFN-beta inhibits IL-12 production by DCs over unknown mechanisms. This study was designed to investigate in vitro effects of GA and IFN-beta on the development and function of DCs from MS patients and healthy controls, and to explore their possible synergistic effects on DCs. DCs were generated from adherent blood mononuclear cells (MNCs). GA or IFN-beta or both, when added at initiation of DC cultures, rapidly promoted the development of adherent MNCs into floating, activated DCs as reflected by up-regulation of HLA-DR and CD86 expression. IFN beta, but not GA, induced IL-3R expression on DCs. Compared to DCs from healthy controls, MS patients' DCs expressed higher levels of the myeloid DC marker CD1a and produced lower amounts of IL-10. GA reduced IL-12 production by DCs. IFN-beta also reduced IL-12, but increased IL-10 production by DCs from both MS patients and healthy controls. GA and IFN-beta synergistically suppressed CD1a and enhanced CD86 expression on MS DCs. These findings document novel mechanisms of action of GA and IFN-beta at the DC level in MS. PMID- 11730947 TI - IL-7-enhanced T-cell response to myelin proteins in multiple sclerosis. AB - In this study, we investigated the in vitro proliferative response of peripheral blood T lymphocytes from MS patients and controls to MBP and MOG either in the absence or in the presence of the conditioning factor IL-7. In the absence of IL 7, T-cell reactivity to MOG and MBP was similar in MS patients and controls even if an increased MBP response was found in a subgroup of patients with active disease. In the presence of IL-7, increased T-cell reactivity to MBP was observed in MS patients suggesting that their MBP-specific T cells are in a different functional state. PMID- 11730948 TI - Immunoglobulin isotypes reveal a predominant role of type 1 immunity in multiple sclerosis. AB - IgG, its subclasses and IgE concentrations were measured in cerebrospinal fluid (CSF) and serum of multiple sclerosis (MS) patients and matched controls as surrogate markers for type 1 and type 2 immunity. IgE indices were significantly reduced in MS patients compared to controls. In contrast, IgG1 was elevated in CSF of MS patients and elevated indices indicated intrathecal synthesis. Because isotype switching to IgE and IgG4 is driven by type 2 immunity and occurrence of IgG1 has previously been found in type 1 immunity-dominated diseases, the results underscore a role of type 1 immunity in MS. PMID- 11730949 TI - Peripheral blood lymphocytes muscarinic cholinergic receptor subtypes in Alzheimer's disease: a marker of cholinergic dysfunction? AB - Muscarinic M2-M5 muscarinic cholinergic receptors were investigated in peripheral blood lymphocytes of patients with mild cognitive impairment of the Alzheimer's type (MCIAT), probable Alzheimer's disease (AD) and probable vascular dementia (VaD). [3H]-N-methyl scopolamine (NMS) in the presence of muscarinic antagonists and Mamba venom to occlude different receptor subtypes was used as radioligand. Analysis of [3H]-NMS binding curves without receptor subtype assessment resulted in a slight decrease of receptor density in AD patients. Evaluation of receptor subtypes in MCIAT and AD patients revealed a decrease of M3 receptor by more than 50%, an increase of M4 receptor expression by about 20% and no changes of M2 or M5 receptors. The expression of M2-M5 receptors was unaltered in VaD patients. Strong positive and negative correlations respectively were found between the density of lymphocyte M3 and M4 receptors and MMSE score in both MCIAT (0.78 for M3 receptor and 0.80 for M4 receptor) and AD (0.82 for M3 receptor and 0.83 for M4 receptor) patients. These findings suggest that changes in the expression of peripheral blood lymphocyte M3 and M4 receptors in AD are related to the degree of cognitive impairment. Assessment of lymphocyte muscarinic receptor subtypes may contribute to characterization of cholinergic impairment in AD. PMID- 11730951 TI - The plasma membrane estrogen receptor: nuclear or unclear? PMID- 11730952 TI - A role for neuronal K(ATP) channels in metabolic control of the seizure gate. PMID- 11730954 TI - Quenching fire with fat: phosphatidylinositides as putative regulators of pain. PMID- 11730955 TI - Developments in whole-body molecular imaging of live subjects. AB - New molecular imaging technologies are being developed specifically for imaging animal models of human disease. Positron emission tomography (PET) in particular allows in vivo biochemistry to be studied with a high degree of sensitivity and specificity, and provides direct in vivo information on molecular and cellular pathways that underlie disease mechanisms and therapeutics. However, clinical PET systems have inadequate resolution for imaging small animals. Thus, specialized high-resolution PET hardware and software are now being developed. PMID- 11730956 TI - Localization of tamoxifen-induced DNA adducts in rat liver. PMID- 11730957 TI - OMInously, a mitochondrial serine protease is now unveiled as the third 'MAC'. PMID- 11730970 TI - Comparing nuclear receptors in worms, flies and humans. AB - Complete nucleotide sequences are now available for different species of the animal kingdom: Caenorhabditis elegans - a nematode, Drosophila - an insect, and humans - a mammal. Such information makes it possible to compare the set of nuclear receptors found in these organisms, and to discuss the possible reasons for the differences observed. The human genome sequencing identified few new receptors, which implies that most nuclear receptors have now been found. However, information about polymorphisms and regulating sequences, obtained through genomic sequencing, will be important for understanding receptor function and disease mechanisms. The surprisingly large number of nuclear receptors in C. elegans might have implications for the development of pharmaceuticals and the understanding of the function of these animals. By contrast, Drosophila has few nuclear receptors; however, examination of the unique nuclear receptors provides information about the function of these receptors. PMID- 11730971 TI - Modelling of promiscuous receptor-Gi/Gs-protein coupling and effector response. AB - A single G-protein-coupled receptor might activate multiple G-protein species. This multiplex coupling ability can be used by tissues to regulate signalling; for the pharmacologist, such multiplex coupling might cause difficulties in the interpretation of experimental data. In this article, we present mathematical models for the activation of two separate G-protein species by a single receptor. Issues addressed concern mutual antagonism between the G proteins and the availability of an already activated receptor for interaction with a new G protein (receptor-G-protein-effector complexing versus free diffusion of G proteins) in addition to receptor-G-protein precoupling at different G-protein and receptor expression levels. The output from the receptor models uses, as readout, a new model for adenylyl cyclase regulation by two allosteric regulators (i.e. G(s) and G(i)). PMID- 11730972 TI - Evidence for epsilon-opioid receptor-mediated beta-endorphin-induced analgesia. AB - Among the opioid receptors, which have been pharmacologically classified as mu, delta, kappa and epsilon, the existence of the epsilon receptor has been controversial, and this receptor is generally not recognized as a member of the opioid peptide receptor family because it has not been precisely characterized. However, results from pharmacological, physiological and opioid receptor binding studies clearly indicate the presence of epsilon-opioid receptors, which are distinct from mu-, delta- or kappa-opioid receptors. This putative epsilon-opioid receptor is stimulated supraspinally by the endogenous opioid peptide beta endorphin, which induces the release of Met-enkephalin, which, in turn, acts on spinal delta2-opioid receptors to produce antinociception. In this article, this beta-endorphin-sensitive epsilon-opioid receptor-mediated descending pain control system, which is distinct from that activated by the mu-opioid receptor agonist morphine, is described and the physiological role of the beta-endorphin-mediated system in pain control activated by cold-water swimming and intraplantar injection of formalin is discussed. PMID- 11730973 TI - Cerebrovascular structure and dementia: new drug targets. AB - Effective pharmacological treatment of cognitive disorders in dementia is lacking despite extensive efforts to produce active therapy aimed at neuronal and vascular targets. In this review, the evidence for the involvement of vascular mechanisms in the pathology and evolution of dementia will be examined and the potential importance of age-related changes in cerebrovascular structure and cerebral blood flow (CBF) autoregulation will be discussed. With a description of recent clinical results (on statins, angiotensin-converting enzyme inhibitors and Ca(2+) channel blockers) and experimental results (on beta-amyloid), the impact of drugs on cerebrovascular targets is examined. The working hypothesis that targeting vascular mechanisms in dementia is an option for future therapy is proposed. PMID- 11730974 TI - NMDA receptor antagonists as analgesics: focus on the NR2B subtype. AB - Ifenprodil and a group of related compounds are selective antagonists of NR2B containing NMDA receptors. These compounds are antinociceptive in a variety of preclinical pain models and have a much lower side-effect profile compared with other NMDA receptor antagonists. It remains unclear whether the improved safety of these compounds is due to their subtype selectivity or to a unique mode of inhibition of the receptor. Human trials have so far confirmed the good tolerability of these subtype-selective NMDA receptor antagonists; however, whether they are as effective as other NMDA receptor antagonists in pain patients remains to be demonstrated. PMID- 11730975 TI - Universality of receptor channel responses. AB - Rate parameters estimated for neurotransmitter-gated receptor channel opening and receptor desensitization are classified according to their dependence on the temporal resolution of the techniques applied in the measurements. Because allosteric proteins constituting receptor channels impose restrictions on the types of model suitable to describe the dynamic response of channels to neurotransmitters, Markovian, non-linear or fractal dynamic models and their possible extension to receptor channel response in excitable membranes are discussed. PMID- 11730978 TI - Glutamate release from adult primary sensory neurons in culture is modulated by growth factors. AB - The aim of this study was to examine possible modulatory effects of some trophic molecules, i.e. nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF), on potassium (K(+))-, bradykinin (BK)- or capsaicin (CAPS)-evoked release of glutamate (GLU) from dorsal root ganglion (DRG) neurons in vitro. BK (0.5 and 1 microM) induced a dramatic and significant increase in glutamate release. Neither CAPS nor K(+) (60 mM) produced any significant increase of GLU release vs. basal levels during a 5 min stimulation. The BK-evoked release of GLU was almost completely blocked by HOE 140, a selective BK2-receptor antagonist at high doses. Basal release of GLU was significantly reduced in cultures grown in the presence of bFGF, whereas BDNF and NGF had no significant effect. Incubation with growth factors generally decreased the BK-stimulated GLU release, an effect most pronounced for bFGF, which completely blocked BK-stimulated release. The rise in intracellular [Ca(2+)] following stimulation with BK (100 nM-1 microM), potassium (60 mM) or ATP (10 microM) was also studied using a Ca(2+)-sensitive indicator, Fura-2, in cultures grown in basal medium with or without bFGF. None of the bFGF-treated cells exhibited strong Ca(2+) responses to BK or ATP stimulation, while 10-20% of the responding cells grown in basal medium exhibited strong responses. The K(+) induced increase of [Ca(2+)] did not vary between the different groups. The present findings suggest that sensory neurotransmission involving glutamate may be modulated by growth factors and that regulation of intracellular Ca(2+) homeostasis may be a contributing factor. PMID- 11730979 TI - Vasorelaxant effect of glucagon-like peptide-(7-36)amide and amylin on the pulmonary circulation of the rat. AB - The gastrointestinal peptides glucagon-like peptide-1(7-36)amide (GLP-1) and amylin are currently being tested in clinical trials for the treatment of diabetes mellitus due to their effects in lowering blood glucose. Receptors for these polypeptides also exist in the lung and since polypeptides are known to modulate airway and pulmonary vascular tone, we investigated whether GLP-1 and amylin act similarly in the lung. We compared their effects with the well-known actions of calcitonin gene-related peptide (CGRP) and vasoactive intestinal peptide (VIP). Both GLP-1 and amylin induced a dose-dependent and time-reversible endothelial-dependent relaxation of preconstricted pulmonary artery rings. Amylin was approximately as strong as VIP and CGRP, GLP-1 however, was 2.3-fold less potent. GLP-1 as well as amylin also reduced the vascular tone in the isolated, perfused and ventilated rat lung. In contrast to their action on the pulmonary vasculature, neither GLP-1 nor amylin showed any effect on the tone of isolated preconstricted trachea rings. In conclusion, GLP-1 and amylin represent two additional peptides which may modulate pulmonary vascular tone. PMID- 11730980 TI - Pituitary aminopeptidase activities involved in blood-pressure regulation are modified by dietary cholesterol: sex differences. AB - Given that the existence of a local renin-angiotensin system (RAS) in the pituitary and its participation in the regulation of blood pressure and other biological functions are widely accepted, the aim of this work is to analyze the influence of dietary cholesterol on the activity of the enzymes involved in the metabolism of the effector peptides of the renin-angiotensin system (angiotensin II and III) and vasopressin, in the pituitary of male and female mice fed on a cholesterol-enriched diet (1% cholesterol and 0.5% cholic acid). Soluble and membrane-bound pituitary aminopeptidase A (aspartyl- and glutamyl aminopeptidase), aminopeptidase M (alanyl-aminopeptidase), aminopeptidase B (arginyl-aminopeptidase) and cystinyl-aminopeptidase activities were fluorimetrically measured. In female mice, cholesterol-enriched diet produced a significant increase in soluble aspartyl- and membrane-bound aspartyl- and glutamyl-aminopeptidase activities, and a significant decrease in membrane-bound alanyl-, arginyl- and cystinyl-aminopeptidase activities. In male mice, after feeding the diet, a significant increase in soluble glutamyl- and membrane-bound arginyl-aminopeptidase activities was observed. Our results indicate differential effects of dietary cholesterol on the metabolism of angiotensin II and III and vasopressin in the pituitary of male and female mice. PMID- 11730981 TI - The exocytotic signaling pathway induced by nerve growth factor in the presence of lyso-phosphatidylserine in rat peritoneal mast cells involves a type D phospholipase. AB - Nerve growth factor (NGF) has been previously shown to induce exocytosis in rat peritoneal mast cells (RPMCs) in the presence of lyso-phosphatidylserine (lysoPS) by interacting with high-affinity NGF receptors of the TrkA-type. In RPMCs, type D phosphatidylcholine-selective phospholipases (PLDs) have been postulated to be involved in some exocytotic signaling pathways induced by different agonists. The aim of the present study was to assess a putative functional role of PLD for NGF/lysoPS-induced exocytosis in RPMCs. In 1-[14C]palmitoyl-2-lyso-3 phosphatidylcholine-labelled RPMCs, NGF/lysoPS stimulated the formation of diacylglycerol (DAG) and, in the presence of ethanol (1% [v/v]), phosphatidylethanol (PEtOH). These data indicate PLD-activation by NGF/lysoPS in RPMCs. Preincubation of RPMCs for 2 min with ethanol, an inhibitor of PLD-derived DAG-formation, dose-dependently (IC(50): 0.6% [v/v]) and agonist-selectively inhibited the NGF/lysoPS induced release of [3H]serotonin ([3H]5-HT) in [3H]5-HT loaded RPMCs, confirming the functional importance of PLD-action. Exocytosis and PEtOH-production was potently inhibited by the broad-spectrum serine/threonine kinase inhibitor staurosporine and activated by the protein kinase C(PKC) activator PMA (phorbol-12-myristate-13-acetate) suggesting a role for PKC as mediator for NGF/lysoPS-induced activation of PLD. PMID- 11730982 TI - Cellular expression of CCK-A and CCK-B/gastrin receptors in human gastric mucosa. AB - Gastrin stimulates gastric acid secretion in various species, but the role of the structurally related CCK for the peripheral regulation of acid secretion in humans remains controversial. Moreover, species differences in CCK receptor function and expression have been reported. We therefore sought to identify the cellular targets of CCK and gastrin within the human gastric mucosa in situ. Gastric biopsies were collected from 15 patients without gastric disease. Expression of CCK receptor subtypes was detected in individual cells of the gastric mucosa by reverse transcription (RT)-PCR in situ, immunohistochemistry and confocal laser scanning microscopy, using antisera against the CCK-A or CCK B/gastrin receptor subtype. Both CCK-A and CCK-B receptors were detected in antral and oxyntic mucosa at the mRNA and protein level. In fundic mucosa, CCK-A receptor mRNA and protein mapped to D cells (37.4+/-7.7). Besides, individual chief cells, mucous neck cells and parietal cells (12.3+/-4.7%) expressed CCK-A receptors. CCK-B/gastrin receptor mRNA and protein were detected in parietal cells (57.4+/-11.1%) and in neuroendocrine cells (33.2+/-4.4%) expressing chromogranin A. Furthermore, epithelial cells within the neck of the gastric gland were found to express the CCK-B/gastrin receptor. We conclude that (i) identification of CCK-A receptors on somatostatin producing D cells in humans provide the anatomical basis for a receptor-mediated mode of action of CCK on somatostatin release and (ii) detection of either CCK receptor subtype in the putative stem cell compartment implies a role of CCK in the maintenance of tissue homeostasis in human gastric mucosa. PMID- 11730983 TI - Synthesis and characterization of tetramethylrhodaminethiocarbamoyl-(Glu(1)) epidermal mitosis-inhibiting pentapeptide. AB - A fluorescent analog of epidermal mitosis-inhibiting pentapeptide (pGlu-Glu-Asp Ser-Gly) was synthesized by reacting tetramethylrhodamine isothiocyanate with ring-opened epidermal mitosis-inhibiting pentapeptide. The ring-opening reaction of the pyrrolidone moiety was performed with mild acidic hydrolysis and the product purified by reversed-phase high-performance liquid chromatography. Tetramethylrhodaminethiocarbamoyl-(Glu(1))-epidermal mitosis-inhibiting pentapeptide was purified by chromatography on Sephadex G-25 and reversed-phase high-performance liquid chromatography. After characterization by amino acid analysis, the analog was incubated in presence of A431 cell line to visualize the cellular localization of the epidermal mitosis-inhibiting pentapeptide. The data gave negative results. PMID- 11730984 TI - Role of tachykinin receptors in the central processing of afferent input from the acid-threatened rat stomach. AB - Noxious challenge of the rat gastric mucosa by hydrochloric acid (HCl) is signalled via vagal afferent neurons to several brain nuclei in which tachykinins and tachykinin receptors are present. Therefore, we tested whether tachykinin receptor antagonists would modify the central transmission of input from the acid threatened stomach. Neuronal excitation was visualized by in situ hybridization autoradiography (ISH) of c-fos messenger ribonucleic acid (mRNA) 45 min after intragastric (IG) administration of HCl (0.5 M; 10 ml/kg). This stimulus has previously been shown to cause neurons in the nucleus tractus solitarii (NTS), lateral parabrachial nucleus (LPB), paraventricular (Pa) nuclei, supraoptic (SO) nucleus, central amygdala (CeA), area postrema (AP), subfornical organ (SFO) and habenula (Hb) to express c-fos mRNA. Intraperitoneal (IP) pretreatment with the NK1 receptor antagonist GR-205,171 (3 mg/kg) attenuated the acid-induced transcription of c-fos mRNA in NTS and augmented it in SFO. The NK2 receptor antagonist SR-144,190 (0.1 mg/kg, IP) had no effect. Subcutaneous administration of the NK3 receptor antagonist SB-222,200 (20 mg/kg) reduced the c-fos mRNA response in AP and SFO and enhanced it in Hb. These data show that the transmission of input from the acid-threatened stomach in distinct brain nuclei involves tachykinins acting at NK1 and NK3 receptors, but not NK2 receptors. PMID- 11730985 TI - Centrally applied atrial natriuretic factor diminishes bile secretion in the rat. AB - Little is known about the role of centrally applied peptides in the regulation of bile secretion. We previously reported that the intravenous injection of atrial natriuretic factor (ANF) reduces bile acid dependent flow without affecting portal venous pressure in the rat. In the present work, we studied the effects of centrally applied ANF on bile secretion and the possible pathways involved. Rats were cannulated in the brain lateral ventricle for the administration of 1, 10 and 100 ng/microl ANF. After 1 week, the common bile duct was cannulated and bile samples were collected every 15 min for 60 min after the administration of ANF. The excretion rate of various biliary components was assessed. Bile secretion experiments were also performed after bilateral truncal vagotomy or atropine administration to evaluate the participation of a vagal pathway. In addition, the role of the sympathetic system was addressed by combined administration of propranolol and phentolamine. Centrally applied ANF did not modify blood pressure but diminished bile flow and bile acid output. It also reduced sodium and potassium secretion but did not modify protein or phospholipid excretion. Neither bilateral truncal vagotomy nor atropine administration abolished ANF response. Furthermore, combined administration of adrenergic antagonists did not alter ANF inhibitory effect on bile flow. In conclusion, centrally applied ANF reduced bile acid dependent flow not through a vagal or adrenergic pathway in the rat, suggesting the involvement of a peptidergic pathway. PMID- 11730986 TI - Up-regulation of splenic prohormone convertases PC1 and PC2 in diabetic rats. AB - Organisms respond to infection in a complex manner involving bidirectional interactions between the neuroendocrine and immune systems. Many of the bioactive endocrine/immune factors are synthesized in a precursor form and are expected to be activated by prohormone convertases (PCs). Since patients with both type 1 and type 2 diabetes have an increased incidence and severity of infections, we hypothesized that in a condition of hyperglycemia, these processing enzymes would be activated in an immune tissue, the spleen. To test this hypothesis, we treated rats with intraperitoneal streptozotocin (STZ) (50 mg/kg/day) daily for 5 days and measured splenic PC1 and PC2 mRNA by ribonuclease protection assay. We found that PC1 mRNA was increased 6.0+/-0.02-fold (P<0.05) and PC2 mRNA was increased 1.80+/-0.01-fold (P<0.005) in the spleen of rats that received STZ compared to rats that received vehicle. Western blot indicated that the 75-kDa form of PC1 was the only form of PC1 present in the spleen and that this form increased with STZ treatment. Immunohistochemistry revealed that PC1 was found in both the white pulp (T-lymphocytes) and red pulp (monocytes and macrophages) and that its increase in immunoreactivity occurred primarily in the white pulp. PC2 and pro opiomelanocortin (POMC, a possible splenic substrate for PC1/PC2) immunoreactivity was found predominantly in the red pulp. STZ induced an increase in splenic PC1 and POMC, but not PC2 protein levels. We conclude that in the STZ model of diabetes, splenic PCs are induced, which could lead to an increased activation of many immune-derived hormones. We speculate that this up-regulation of prohormone converting enzymes may be related to the increased infections seen in patients with both type 1 and type 2 diabetes. PMID- 11730987 TI - A role for the angiotensin AT4 receptor subtype in overcoming scopolamine-induced spatial memory deficits. AB - There is increasing interest in the role of the brain angiotensin AT4 receptor subtype in cognitive processing. This receptor subtype is activated by angiotensin IV (AngIV), is heavily distributed in the mammalian hippocampus, neocortex, and cerebellum, and has been linked with a learning and memory function. The present investigation utilized intracerebroventricular (i.c.v.) infused scopolamine hydrobromide (scop), a muscarinic receptor antagonist, to disrupt acquisition of the circular water maze task of spatial memory. All animals received 2 days of training trials (five trials/day) using a visible platform in an effort to preclude subsequent confounding by scopolamine-induced sensory and/or motor impairments. In the first experiment, i.c.v.-infused scopolamine (70 nmol) was followed by 0, 10, 100, or 1000 pmol i.c.v. doses of Nle(1)-AngIV in separate groups of rats. Results indicated that each dose of Nle(1)-AngIV improved the poor acquisition of this task induced by scopolamine treatment. However, the 100- and 1000-pmol doses were most effective with respect to latency and distance to find the submerged pedestal. A second experiment demonstrated that treatment with a specific AT4 receptor antagonist, Nle(1), Leual(3)-AngIV (1000 pmol), blocked the ability of Nle(1)-AngIV (100 pmol) to improve the performance of scopolamine-compromised rats. These results support the notion that hippocampal AT4 receptors are involved in spatial memory processing, and that activation of these binding sites can overcome the disruption of spatial memory accompanying treatment with a muscarinic receptor antagonist. PMID- 11730988 TI - PACAP mediates the neural proliferative pathway of Mastomys enterochromaffin-like cell transformation. AB - BACKGROUND AND AIM: Pituitary adenylate-cyclase activating peptide (PACAP) is a more potent proliferative agent than gastrin for rat enterochromaffin-like (ECL) cell proliferation in vitro. The role of this neurotransmitter during gastrin mediated ECL cell tumor formation and gastrin-autonomous ECL cell neoplasia is unknown. METHODS AND RESULTS: ECL cell transformation was induced in the Mastomys using 16 wk H2 receptor blockade of acid inhibition. Examination of the epithelial fundic mucosa demonstrated that PACAP-immunoreactivity significantly increased in the tumor mucosa compared to the naive stomach, and was associated with ECL cells. Naive and tumor ECL cells were then purified (approximately 95%) from Mastomys and the presence of all three PACAP/VPAC receptor subtypes was demonstrated by polymerase chain-reaction amplification. Thereafter, cells were maintained in short-term (48 h) primary cultures. PACAP significantly (p<0.05) increased 24 h bromo-deoxyuridine uptake (approximately 4-fold) in both cell types with estimated EC(50) values of approximately 4x10(-16) M and approximately 2x10(-16) M, respectively. Specific receptor antagonists (PAC1/VPAC1) of PACAP competitively inhibited these proliferative effects in naive cells. Oligonucleotide antisense directed against PAC1 significantly inhibited PACAP stimulated DNA synthesis by approximately 85% (p<0.05) in tumor cells. CONCLUSION: PACAP is a potent and effective modulator of ECL cell proliferation. The expression of this neuropeptide and its receptors, particularly PAC1, suggest the existence of a neural regulatory pathway of ECL cell proliferation and transformation. PMID- 11730990 TI - Late morbidity: the Damocles Sword of radiotherapy? PMID- 11730991 TI - Consequential late effects in normal tissues. AB - Unconventional, more aggressive irradiation protocols are usually associated with aggravation of acute reactions. In recent clinical studies, this has resulted in modulation of late effects in the same organ. This phenomenon has been termed consequential late effect (CLE). Correlations between acute and late effects have been reported in a number of tissues. Moreover, some radiobiological parameters may be used to differentiate between consequential and generic late effects: Dose fractionation and overall treatment time have a similar effect on acute and consequential responses, but opposing effects on generic late effects. Modulation of acute effects will affect the consequential component of late sequelae. Similarly, it will be influenced by the irradiated volume if a volume effect exists for the acute response. Moreover, markers for the acute response should be predictive for consequential effects. The present review gives preclinical and clinical evidence for CLE. These are predominantly found in organ systems where the acute response (of the epithelial lining) is associated with an impairment of the barrier against mechanical or chemical stress, which may cause additional trauma to the underlying tissues. Therefore, CLE are mainly found in the urinary and intestinal system, in mucosa and, to some extent, in skin. In these tissues with a consequential component of the late sequelae, amelioration of the acute response to irradiation may be a useful approach to minimize late side effects of effective radiation therapy. PMID- 11730992 TI - Quantification of late complications after radiation therapy. AB - BACKGROUND: An increasing number of patients survive cancer after having received radiation therapy. Therefore, the occurrence of late normal tissue complications among long-term survivors is of particular concern. METHODS: Sixty-three patients treated by radical surgery and irradiation for rectal carcinoma were subjected to an unconventional sandwich therapy. Preoperative irradiation was given in four fractions of 5 Gy each applied within 2 or 3 days; postoperative irradiation consisted mostly of 15 x 2 Gy (range, 20-40 Gy). A considerable proportion of these patients developed severe late complications (Radiother Oncol 53 (1999) 177). The data allowed a detailed analysis of complication kinetics, leading to a new model which was tested using data from the literature. RESULTS: Data on late complications were obtained for eight different organs with a follow-up of up to 10 years. For the various organs, the percentage of patients being free from late complications, plotted as a function of time after start of radiation therapy, was adequately described by exponential regression. From the fit, the parameter p(a) was obtained, which is the percentage of patients at risk in a given year of developing a complication in a given organ during that year. The rate p(a) remained about constant with time. Following sandwich therapy, the annual incidence of complications in the bladder, ileum, lymphatic and soft tissue, and ureters was about the same (p(a)=10-14%/year), whereas complications in bone or dermis occurred at lower rates (4.7 or 7.5%/year, respectively). DISCUSSION: Numerous data sets collected from published reports were analyzed in the same way. Many of the data sets studied were from patients in a series where there was a high incidence of late effects. Three types of kinetics for the occurrence of late effects after radiotherapy were identified: Type 1, purely exponential kinetics; Type 2, exponential kinetics, the slope of which decreased exponentially with time; Type 3, curves composed of two components, a fast initial decline followed by an exponential decrease. For each kind of kinetics, provided that the dose distribution is not too heterogeneous, the incidence of late effects appears to occur at exponential or approximately exponential kinetics, even many years after treatment. This implies that a random process might be involved in the occurrence of late radiation sequelae. CONCLUSIONS: There might be a lifelong risk of developing late complications, of which patients and clinicians should be aware. It appears worthwhile to try to identify, in follow-up examinations of patients after radiation therapy, what kind of processes might be involved in triggering subclinical residual injury to develop into a clinically manifest late effect. PMID- 11730993 TI - Nasopharyngeal carcinomas: analysis of patient, tumor and treatment characteristics determining outcome. AB - PURPOSE: The present study reviews the experience in treatment of 447 patients with nasopharyngeal carcinomas, analyzing patient, tumor and treatment characteristics determining outcome. MATERIALS AND METHODS: There were 322 males and 125 females, their ages ranging from 7 to 85 years (median, 45 years). Two hundred and seventy-two patients had World Health Organization (WHO) type 3 carcinomas, 123 patients had T4 tumors and 320 patients had metastatic cervical lymph nodes. Three-hundred and eight patients were treated with radiation therapy alone and 139 patients with chemotherapy in combination with radiation therapy. Cumulative radiation dose to primary tumor ranged from 50 to 76Gy (median, 70Gy) and radiation dose to metastatic cervical lymph nodes ranged from 46 to 74Gy (median, 66Gy). RESULTS: Follow-up ranged from 0.1 to 19.5 years (mean, 7.6 years). Local complete response was achieved in 357 patients. In multivariate analysis, T-classification, cumulative radiation dose to primary tumor and treatment with chemotherapy in combination with radiation therapy predicted local response. Nodal complete response was achieved in 272 patients. In multivariate analysis, N-classification and radiation dose to metastatic cervical lymph nodes predicted nodal response. Local failure was observed in 70 patients, nodal failure in 35 patients and systemic failure in 114 patients. Overall survival, disease-free survival and disease-specific survival were 33, 32 and 37%, respectively, at 10 years. In multivariate analysis, age, T-classification, N classification, radiation dose and treatment with chemotherapy in combination with radiation therapy predicted overall survival whereas T-classification, N classification, radiation dose and treatment with chemotherapy in combination with radiation therapy predicted both disease-free survival and disease-specific survival. CONCLUSIONS: Radiation therapy alone appears to be an adequate and viable treatment for patients with early-stage nasopharyngeal carcinomas, whereas treatment with chemotherapy in combination with radiation therapy appears to improve outcome for patients with advanced-stage nasopharyngeal carcinomas. PMID- 11730995 TI - Combined chemotherapy and radiation versus radiation alone in the management of localized angiocentric lymphoma of the head and neck. AB - BACKGROUND AND PURPOSE: To clarify the clinical benefit derived from the combined modality therapy (CMT) comprised of chemotherapy and involved-field radiotherapy (XRT) for stage I and II angiocentric lymphomas of the head and neck. MATERIAL AND METHODS: Of 143 patients with angiocentric lymphoma of the head and neck treated at the Yonsei Cancer Center between 1976 and 1995, 104 patients (XRT group) received involved-field XRT alone with a median dose of 50.4 Gy (range: 20 70 Gy), while 39 patients (CMT group) received a median three cycles (range: 1-6 cycles) of chemotherapy before starting involved-field XRT. The response rate, patterns of failure, complications, and survival data of the XRT group were compared with those of the CMT group. RESULTS: Despite a higher response rate, local failure was the most common pattern of failure in patients of the both groups. The patterns of failure, including the systemic relapse rate were not influenced by the addition of combination chemotherapy. Although both modalities were well tolerated by the majority of patients, aberrant immunologic disorders or medical illnesses, such as a hemophagocytic syndrome, sepsis, intractable hemorrhage, or the evolution of second primary malignancies were more frequently observed in patients of the CMT group. The prognosis of patients in the XRT group was relatively poor, with a 5-year overall actuarial survival rate of 38% and disease-free survival rate of 32%, respectively. However, their clinical outcome was not altered by the addition of systemic chemotherapy. Achieving complete remission was the most important prognostic factor on univariate and multivariate analyses, but treatment modality was not found to be a prognostic variable influencing survival. CONCLUSIONS: Involved-field XRT alone for angiocentric lymphoma of the head and neck was insufficient to achieve an improved survival rate, but the combination of chemotherapy and involved-field XRT failed to demonstrate any therapeutic advantage over involved-field XRT alone. PMID- 11730994 TI - Alternating chemoradiotherapy for nasopharyngeal cancer using cisplatin and 5 fluorouracil: a preliminary report of phase II study. AB - Alternating chemoradiotherapy was performed in 35 patients with locally advanced nasopharyngeal cancer. The median duration of follow-up was 20 months, and the 2 year progression free and overall survival rates were 83% (95% confidence interval: 66~101%) and 94% (95% CI: 84~105%), respectively. This method may be useful in treating nasopharyngeal cancer. PMID- 11730996 TI - Parotid and submandibular/sublingual salivary flow during high dose radiotherapy. AB - It was studied whether differences in acute radiosensitivity exist between parotid and submandibular/sublingual glands. The results revealed that salivary flow rates decreased dramatically during the first 2 weeks of radiotherapy. Neither recovery nor significant differences were observed between the production of saliva from the parotid and submandibular/sublingual glands during the 13 weeks observation period. PMID- 11730997 TI - Intensity-modulated radiation therapy reduces late salivary toxicity without compromising tumor control in patients with oropharyngeal carcinoma: a comparison with conventional techniques. AB - BACKGROUND AND PURPOSE: Intensity-modulated radiation therapy (IMRT) offers superior dosimetric conformity for normal tissue sparing in patients with oropharyngeal cancer. In this study, acute and late toxicity, and tumor control were compared between conventional beam arrangement (CRT) and IMRT. MATERIALS AND METHODS: Between January 1970 and December 1999, 430 patients with carcinoma of the oropharynx were treated at the Mallinckrodt Institute of Radiology. There were 260 patients with tonsil primary tumors and 170 patients with tumors arising from the base of the tongue. Twenty-four (6%) patients had stage I disease, 88 (20%) had stage II, 128 (30%) had stage III, and 190 (44%) had stage IV disease. Patients were divided into five treatment groups. Group I consisted of 109 patients who received preoperative CRT. Group II consisted of 142 patients who received postoperative CRT. Group III consisted of 153 patients who received definitive CRT. Inverse planning IMRT (Peacock, NOMOS) was used to treat 14 patients postoperatively (Group IV) and 12 patients definitively without surgery (Group V). Acute and late normal tissue side-effects were scored according to the Radiation Therapy Oncology Group radiation morbidity criteria. The median follow up was 3.9 years. RESULTS: The 2-year local-regional control values for the five studied groups were 78, 76, 68, 100 and 88%, respectively. The 2-year disease free survival values for the five studied groups were 68, 74, 58, 92 and 80%, respectively. IMRT significantly reduced the incidence of late xerostomia. CONCLUSIONS: When IMRT was compared with conventional techniques, the dosimetric advantage of IMRT did translate into a significant reduction of late salivary toxicity in patients with oropharyngeal carcinoma. No adverse impact on tumor control and disease-free survival was observed in patients treated with IMRT. PMID- 11730998 TI - Evaluation of a leaf position optimization tool for intensity modulated radiation therapy of head and neck cancer. AB - BACKGROUND AND PURPOSE: Since 1996, patients are treated at Ghent University Hospital with a multi-segment technique using MultiLeaf Collimators. The segments were obtained by using the Beam's eye view projections of the planning target volume (PTV) and the organs at risk (OARs), after which the segments weights were optimized. To investigate if optimization of the leaf positions would further improve the intensity modulated radiation therapy (IMRT) plans, a tool optimizing leaf positions and segment weights simultaneously, was developed. This tool is called SOWAT, which is the acronym for segment outline and weight adapting tool. MATERIAL AND METHODS: The tool evaluates the effects of changing the position of each collimating leaf of all segments on the value of the objective function. Only changes that improve the value of the objective function are retained. Between December 1999 and January 2001, 30 head and neck patients were treated with IMRT. Two patient groups were distinguished: pharyngeal and laryngeal tumors (n=17) and sinonasal tumors (n=13). A specific set of physical endpoints was evaluated for each group. Dose statistics of the treatment plans without and with SOWAT were analyzed. RESULTS: When using SOWAT for the pharyngeal and laryngeal cases, the PTV dose homogeneity increased with a median of 11% (range 2-27%), while the maximum dose to the spinal cord was decreased for 14 of the 17 patients. In four plans where parotid function preservation was a goal, the parotid mean dose was lower than 26 Gy in one plan without SOWAT, and in four plans with SOWAT. For the sinonasal tumors, the PTV dose homogeneity increased with a median of 7% (range 1-14%). SOWAT lowered the mean dose to 53 of the 63 optic pathway structures (retina, optic nerve and optic chiasm). SOWAT leaves the number of segments unchanged and has little or no effect on the delivery time. CONCLUSIONS: SOWAT is a powerful tool to perform the final optimization of IMRT plans, without increasing the complexity of the plan or the delivery time. PMID- 11730999 TI - A treatment planning comparison of 3D conformal therapy, intensity modulated photon therapy and proton therapy for treatment of advanced head and neck tumours. AB - BACKGROUND AND PURPOSE: In this work, the potential benefits and limitations of different treatment techniques, based on mixed photon-electron beams, 3D conformal therapy, intensity modulated photons (IM) and protons (passively scattered and spot scanned), have been assessed using comparative treatment planning methods in a cohort of patients presenting with advanced head and neck tumours. MATERIAL AND METHODS: Plans for five patients were computed for all modalities using CT scans to delineate target volume (PTV) and organs at risk (OAR) and to predict dose distributions. The prescribed dose to the PTV was 54 Gy, whilst the spinal cord was constrained to a maximum dose of 40.5 Gy for all techniques. Dose volume histograms were used for physical and biological evaluation, which included equivalent uniform dose (EUD) calculations. RESULTS: Excluding the mixed photon-electron technique, PTV coverage was within the defined limits for all techniques, with protons providing significantly improved dose homogeneity, resulting in correspondingly higher EUD results. For the spinal cord, protons also provided the best sparing with maximum doses as low as 17 Gy. Whilst the IM plans were demonstrated to be significantly superior to non modulated photon plans, they were found to be inferior to protons for both criteria. A similar result was found for the parotid glands. Although they are partially included in the treated volume there is a clear indication that protons, and to a lesser extent IM photons, could play an important role in preserving organ functionality with a consequent improvement of the patient's quality of life. CONCLUSIONS: For advanced head and neck tumours, we have demonstrated that the use of IM photons or protons both have the potential to reduce the possibility of spinal cord toxicity. In addition, a substantial reduction of dose to the parotid glands through the use of protons enhances the interest for such a treatment modality in cases of advanced head and neck tumours. However, in terms of target coverage, the use of 3D conformal therapy, although somewhat inferior in quality to protons or IM photons, has been shown to be a reasonable alternative to the more advanced techniques. In contrast, the conventional technique of mixed photon and electron fields has been shown to be inferior to all other techniques for both target coverage and OAR involvement. PMID- 11731000 TI - Electronic portal image assisted reduction of systematic set-up errors in head and neck irradiation. AB - PURPOSE: To quantify systematic and random patient set-up errors in head and neck irradiation and to investigate the impact of an off-line correction protocol on the systematic errors. MATERIAL AND METHODS: Electronic portal images were obtained for 31 patients treated for primary supra-glottic larynx carcinoma who were immobilised using a polyvinyl chloride cast. The observed patient set-up errors were input to the shrinking action level (SAL) off-line decision protocol and appropriate set-up corrections were applied. To assess the impact of the protocol, the positioning accuracy without application of set-up corrections was reconstructed. RESULTS: The set-up errors obtained without set-up corrections (1 standard deviation (SD)=1.5-2mm for random and systematic errors) were comparable to those reported in other studies on similar fixation devices. On an average, six fractions per patient were imaged and the set-up of half the patients was changed due to the decision protocol. Most changes were detected during weekly check measurements, not during the first days of treatment. The application of the SAL protocol reduced the width of the distribution of systematic errors to 1mm (1 SD), as expected from simulations. A retrospective analysis showed that this accuracy should be attainable with only two measurements per patient using a different off-line correction protocol, which does not apply action levels. CONCLUSIONS: Off-line verification protocols can be particularly effective in head and neck patients due to the smallness of the random set-up errors. The excellent set-up reproducibility that can be achieved with such protocols enables accurate dose delivery in conformal treatments. PMID- 11731001 TI - Adenovirus-p53 gene therapy in human nasopharyngeal carcinoma xenografts. AB - BACKGROUND: One major challenge to human cancer gene therapy, is efficient delivery of the gene-vector complex. METHODS AND RESULTS: Using two distinct human nasopharyngeal carcinoma (NPC) models, we demonstrate that intra-tumoural (IT) administration of adenoviral-mediated wild-type p53 gene therapy (Ad-p53) caused no greater inhibition of tumour growth as compared to ionizing radiation (XRT) alone. Detailed histologic examination of tumour sections demonstrated that <15% of tumour cells were transduced by IT adv-beta-gal. CONCLUSIONS: This report underscores the importance of developing gene transfer vectors, which can provide therapeutic levels of transgene expression efficiently in solid tumours. PMID- 11731002 TI - Tumor vasculature is targeted by the combination of combretastatin A-4 and hyperthermia. AB - BACKGROUND AND PURPOSE: Combretastatin A-4 disodium phosphate (CA-4) enhances thermal damage in s.c. BT(4)An rat gliomas. We currently investigated how CA-4 and hyperthermia affect the tumor microenvironment and neovasculature to disclose how the two treatment modalities interact to produce tumor response. METHODS: By confocal microscopy and immunostaining for von Willebrand factor, we examined the extent of vascular damage subsequent to CA-4 (50 mg/kg) and hyperthermia (waterbath 44 degrees C, 60 min). The influence on tumor oxygenation was assessed using interstitial pO(2)-probes (Licox system) and by immunostaining for pimonidazole. We examined the direct effect of CA-4 on the tumor cell population by flow cytometry (cell cycle distribution) and immunostaining for beta-tubulin (cytoskeletal damage). RESULTS: Whereas slight vascular damage was produced by CA 4 in the BT(4)An tumors, local hyperthermia exhibited moderate anti-vascular activity. In tumors exposed to CA-4 3 h before hyperthermia, massive vascular damage ensued. CA-4 reduced the pO(2) from 36.1 to 17.6 mmHg (P=0.01) in the tumor base, and tumor hypoxia increased slightly in the tumor center (pimonidazole staining). Extensive tumor hypoxia developed subsequent to hyperthermia or combination therapy. Despite a profound influence on beta-tubulin organization in vitro, CA-4 had no significant effect on the cell cycle distribution in vivo. CONCLUSION: Our results indicate that the anti-vascular activity exhibited by local hyperthermia can be augmented by previous exposure to CA-4. PMID- 11731003 TI - Calcium-dependent translocation of synaptotagmin to the plasma membrane in the dendrites of developing neurones. AB - In neurones, the morphological complexity of the dendritic tree requires regulated growth and the appropriate targeting of membrane components. Accurate delivery of specific supplies depends on the translocation and fusion of transport vesicles. Vesicle SNAREs (soluble N-ethylmaleimide sensitive factor attachment protein receptors) and target membrane SNAREs play a central role in the correct execution of fusion events, and mediate interactions with molecules that endow the system with appropriate regulation. Synaptotagmins, a family of Ca(2+)-sensor proteins that includes neurone-specific members involved in regulating neurotransmitter exocytosis, are among the molecules that can tune the fusion mechanism. Using immunocytochemistry, confocal and electron microscopy, the localisation of synaptotagmin I in the dendrites of cultured rat hypothalamic neurones was demonstrated. Synaptotagmin labelling is concentrated at dendritic branch points, and in microprocesses. Following depolarisation, the N-terminal domain of synaptotagmin was detected at the extracellular surface of the dendritic plasma membrane. The insertion of synaptotagmin in the plasma membrane was elicited by L-type Ca(2+) channel activation and by mobilisation of the internal ryanodine-sensitive Ca(2+)stores. Furthermore, the localisation of L type Ca(2+) channels and of ryanodine receptors, relative to the localisation of synaptotagmin in dendrites, suggests that both Ca(2+) entry and intracellular Ca(2+) stores may contribute to the fusion of dendritic transport vesicles with the membrane. Fusion is likely to involve SNAP-25 and syntaxin 1 as both proteins were also identified in dendrites. Taken together these results suggest a putative regulatory role of synaptotagmins in the membrane fusion events that contribute to shaping the dendritic tree during development. PMID- 11731004 TI - Subcellular localization of presenilin 2 endoproteolytic C-terminal fragments. AB - Mutations in the genes that encode the presenilin 1 and 2 (PS1 and PS2) proteins cause the majority of familial Alzheimer's disease (FAD). Differential cleavage of the presenilins results in a generation of at least two C-terminal fragments (CTFs). An increase in the smaller of these two CTFs is one of the few changes in presenilin processing associated with FAD mutations in both PS1 and PS2. Interestingly, the phosphorylation of PS2 modulates the production of the smaller, caspase-derived PS2 CTF, which indicates that the generation of this fragment is a regulated, physiologic event. To date, there is no data concerning the subcellular distribution of the caspase-derived PS2 CTF. Because this fragment is normally present at levels that are difficult to detect, we have used cell lines in which the production of wild-type or N141I mutant PS2 is controlled by a tetracycline-regulated promoter in order to assess the subcellular localization of the caspase CTF in relation to the larger, constitutive PS2 CTF and to PS2 holoprotein. We have found that when levels of PS2 are low, the constitutive CTF colocalizes with markers consistent with localization in the early Golgi-ER-Golgi intermediate compartment (ERGIC) while the caspase CTF colocalizes with markers for the endoplasmic reticulum (ER). Following induction of wild-type or mutant PS2, when the levels of PS2 are high, the primary localization of the constitutive CTF appears to shift from the early Golgi-ERGIC in addition to the ER. Interestingly, while the induction of wild-type PS2 resulted in the localization of the caspase CTF primarily in the ER, the induction of mutant PS2 resulted in the localization of the caspase CTF to both the ER and the early Golgi-ERGIC. In summary, these data suggest that the two presenilin 2 CTFs have different patterns of subcellular localization and that the N141I PS2 mutation alters the localization pattern of the PS2 caspase fragment. PMID- 11731005 TI - Overexpression of muscle specific kinase increases the transcription and aggregation of acetylcholine receptors in Xenopus embryos. AB - Muscle specific kinase (MuSK) mediates agrin-induced acetylcholine receptor (AChR) aggregation on muscle membrane at the neuromuscular junction (NMJ). To examine whether MuSK enhances NMJ formation during embryonic development in vivo, the level of expression of MuSK was manipulated in Xenopus embryos and the functional consequence at the NMJ was assessed. We found that overexpression of MuSK enhanced the formation of NMJ by increasing the aggregation of AChRs at innervated regions in developing embryos. The area of AChR aggregation increased by approximately 2-fold in MuSK injected embryos during the critical stages of NMJ formation. Interestingly, overexpression of MuSK in Xenopus embryos was found to induce the level of AChR transcript. Deletion of the Kringle domain in the MuSK construct did not attenuate the observed induction of AChR transcription and aggregation. Taken together, our findings provide the first demonstration that increased level of MuSK expression in vivo significantly elevate the aggregation and transcription of AChR at the NMJ in developing Xenopus embryos. PMID- 11731006 TI - Abeta(1-42) and aluminum induce stress in the endoplasmic reticulum in rabbit hippocampus, involving nuclear translocation of gadd 153 and NF-kappaB. AB - Apoptosis may represent a prominent form of neuronal death in chronic neurodegenerative disorders, such as Alzheimer's disease. Although apoptosis under mitochondrial control has received considerable attention, mechanisms used within the endoplasmic reticulum (ER) and nucleus in mediating apoptotic signals are not well understood. A growing body of evidence is emerging from different studies which suggests an active role for the ER in regulating apoptosis. Disturbances of ER function have been shown to trigger two different apoptotic pathways; one involves cross-talk with mitochondria and is regulated by the antiapoptotic Bcl-2, and the second is characterized by the activation of caspase 12. Also, stress in the ER has been suggested to result in the activation of a number of proteins, such as gadd 153 and NF-kappa, and in the downregulation of the antiapoptotic protein, Bcl-2. In the present study, the intracisternal injection in aged rabbits of either the neurotoxin aluminum maltolate or of Abeta(1-42), has been found to induce nuclear translocation of gadd 153 and the inducible transcription factor, NF-kappaB. Translocation of these two proteins is accompanied by decreased levels of Bcl-2 in both the ER and the nucleus. Aluminum maltolate, but not Abeta, induces caspase-12 activation which is a mediator of ER specific apoptosis; this is the first report of the in vivo activation of caspase 12. These findings indicate that the ER may play a role in regulating apoptosis in vivo, and could be of significance in the pathology of neurodegeneration and related disorders. PMID- 11731008 TI - Preconditioning with cortical spreading depression does not upregulate Cu/Zn-SOD or Mn-SOD in the cerebral cortex of rats. AB - Previous studies have demonstrated that preconditioning the brain with cortical spreading depression (CSD) induces tolerance to a subsequent episode of ischemia. In other models of preconditioning, induction of ischemic tolerance has been associated with increased expression of the antioxidant enzyme, superoxide dismutase (SOD). The objective of the present study was to determine whether CSD upregulates Cu/Zn-SOD or Mn-SOD. CSD was induced in one hemisphere by applying 2 M KCl to the frontal cortex in Wistar rats. After 2 or 24 h of recovery, Cu/Zn SOD and Mn-SOD mRNA levels were determined in both hemispheres using Northern blot analysis. In separate rats, Cu/Zn-SOD and Mn-SOD protein levels were determined 24 and 72 h after CSD using Western blot analysis. In addition, total SOD, Cu/Zn-SOD and Mn-SOD enzymatic activities were measured 24 and 72 h after CSD using spectrophotometric and zymographic assays. At the times investigated, no significant differences in mRNA or protein levels for Cu/Zn-SOD or Mn-SOD were observed between the ipsilateral and contralateral cortex. Further, there were no significant differences in Cu/Zn-SOD or Mn-SOD enzymatic activities between the two hemispheres at 24 or 72 h after CSD. In addition, CSD did not alter the activities of Cu/Zn-SOD or Mn-SOD in either hemisphere, relative to those in unoperated animals. Taken together, these results fail to support the hypothesis that CSD-induced tolerance is mediated through the upregulation of Cu/Zn-SOD or Mn-SOD. PMID- 11731007 TI - Rapid phosphorylation of the CRE binding protein precedes stress-induced activation of the corticotropin releasing hormone gene in medial parvocellular hypothalamic neurons of the immature rat. AB - The mechanisms of the molecular and neuroendocrine responses to stress in the immature rat have been a focus of intense investigation. A principal regulator of the these responses in both mature and developing rat is the neuropeptide corticotropin releasing hormone (CRH), and levels of hypothalamic CRH mRNA are enhanced by stress. In vitro, transcription of the CRH gene is governed by binding of the phosphorylated form of cAMP responsive element binding protein (pCREB) to the promoter. Here we tested the hypothesis that rapid, stress-induced CRH transcription occurred during the first two postnatal weeks, and is associated with pCREB expression. The time-course of induction of unedited, heteronuclear CRH RNA (CRH hnRNA) was examined in hypothalamic paraventricular nucleus (PVN) of immature rats subjected to both modest and strong acute stressors using in situ hybridization; pCREB abundance was determined in individual neurons in specific PVN sub-nuclei using immunocytochemistry and unbiased quantitative analysis. CRH hnRNA signal was negligible in PVN of immature rats sacrificed under stress-free conditions, but was readily detectable within 2 min, and peaked at 15 min, in PVN of stressed animals. Enhanced pCREB immunoreactivity was evident within 2 min of stress onset, and was enhanced specifically in stress-responsive, CRH-expressing medial parvocellular neurons. These data support the notion that, already during early postnatal life, stress induces rapid CREB phosphorylation, interaction of pCREB-containing transcription complexes with the CRE element of the CRH gene promoter, and initiation of CRH hnRNA production in stress-responsive neurons of rat PVN. PMID- 11731009 TI - Developmental and functional evidence of a role for Zfhep in neural cell development. AB - The rat Zfhep gene encodes a member of the Zfh family of transcription factors having a homeodomain-like sequence and multiple zinc fingers. We examined expression of Zfhep in the rat forebrain during embryonic and postnatal development. Zfhep mRNA was strongly expressed in the progenitor cells of the ventricular zone around the lateral ventricles on E14 and E16, but showed little expression in cells that had migrated to form the developing cortex. Dual labeling with PCNA demonstrated expression of Zfhep mRNA in proliferating cells. Expression of Zfhep in the ventricular zone decreases during late development as the population of progenitor cells decreases. This pattern is distinctly different from other members of the Zfh family. We also examined the expression of Zfhep protein during retinoic acid-induced neurogenesis of P19 embryonal carcinoma cells. Zfhep is highly expressed in P19 neuroblasts, and expression decreases by the time of morphological neurogenesis. Hence, both P19 cells and embryonic brain demonstrate a loss of Zfhep expression during the transition from proliferating precursor to differentiated neural cells. We investigated a possible link between Zfhep and proliferation by treating human glial cell lines with Zfhep antisense phosphorothioate oligodeoxynucleotides. Two Zfhep antisense oligonucleotides repressed proliferation of either U-138 or U-343 glioblastoma cells more than control oligonucleotides. Based on the expression patterns of Zfhep in vivo and in the P19 cell model of neurogenesis, we suggest that Zfhep may play a role in proliferation or differentiation of neural cells. PMID- 11731010 TI - Pharmacological differences between immunoisolated native brain and heterologously expressed rat alpha4beta2 nicotinic receptors. AB - Native brain and heterologously expressed rat alpha4beta2 nicotinic receptors (in Xenopus oocytes and CV-1 cells) were immunoisolated with the anti-alpha4 antibody mAb 299 and their pharmacological properties were compared using [3H](+/ )epibatidine, the novel N-alkylnicotinium analog N-n-octylnicotinium iodide (NONI), and the ganglionic antagonist trimethaphan (TRM). The equilibrium dissociation constant (K(d)) for [3H](+/-)epibatidine binding to the native and heterologously expressed receptors ranged from 13 to 21 pM. The Hill coefficients for [3H](+/-)epibatidine binding to the native and expressed receptors ranged from 0.8 to 1.1 and were consistent with a single high-affinity site. NONI inhibited 30 pM [3H](+/-)epibatidine binding to the native and expressed receptors with similar potency (IC(50) values of 6-7 microM). However, [3H](+/ )epibatidine dissociated 2-3 times more slowly from the native, than from the expressed receptors and TRM inhibited 30 pM [3H](+/-)epibatidine binding to the native receptors (IC(50) value of 330 microM) less potently than it did to the receptors expressed in oocytes (IC(50) value of 16 microM) or CV-1 cells (IC(50) value of 55 microM). The differences between the native and expressed [3H](+/ )epibatidine dissociation rate constants and IC(50) values for TRM were significant for both host cell types, although the values for the CV-1-expressed receptors were closer to the native ones than were those for the oocyte-expressed receptors. Thus, the epibatidine and trimethaphan binding sites in native and expressed alpha4beta2 receptors appear to have significantly different structural or chemical properties. PMID- 11731011 TI - Increase of tyrosine hydroxylase and dopamine transporter mRNA levels in ventral tegmental area of male mice under influence of repeated aggression experience. AB - Tyrosine hydroxylase (TH) and dopamine transporter (DAT) mRNA levels in the ventral tegmental area (VTA) of midbrain were measured by multiplex RT-PCR in male mice with repeated experience of social victories (winners) and social defeats (losers) in 10 daily agonistic confrontations. Two independent experiments revealed enhanced TH and DAT mRNA levels in VTA of the winners in comparison with the losers and controls (animals after 5 days of individual housing). A positive correlation between DAT and TH mRNA levels was shown. PMID- 11731012 TI - Delayed upregulation of GABA(A) alpha1 receptor subunit mRNA in somatosensory cortex of mice following learning-dependent plasticity of cortical representations. AB - Experience-dependent modifications of cortical representational maps are accompanied by changes in several components of GABAergic inhibitory neurotransmission system. We examined with in situ hybridization to 35S-labeled oligoprobe changes of expression of GABA(A) receptor alpha1 subunit mRNA in the barrel cortex of mice after sensory conditioning training. One day and 5 days after the end of short lasting (3 daily sessions) training an increased expression of GABA(A) alpha1 mRNA was observed at the cortical site where the plastic changes were previously found. Learning associated activation of the cerebral cortex increases expression of GABA(A) receptor mRNA after a short post training delays. PMID- 11731013 TI - Structural requirements of benzodiazepines for the inhibition of pig brain nitric oxide synthase. AB - Nitric oxide synthases (NOS) are heme-containing enzymes which catalyse the oxidation of L-arginine to nitric oxide and L-citrulline. Some nitrogenated compounds have been reported to coordinate with the iron atom from the heme group, thus inhibiting NOS. 1,4-Benzodiazepines are nitrogenated compounds which have many physiological effects such as antianxiety, antiepileptic, hypnotic, and muscle relaxation properties. The aim of this paper was to measure the effect of different benzodiazepines on NOS activity in pig brain extracts. Medazepam, pinazepam, diazepam, oxazepam and alprazolam competitively inhibited NOS with IC(50) in the micromolar range. Other benzodiazepines showed no effect at concentrations as high as 200 microM. Due to the structural similarity of the benzodiazepine ring nucleus with L-arginine, we propose a benzodiazepine-enzyme interaction to explain the competitive inhibitions. By comparing benzodiazepine effects and their structures, the inhibitory effect of benzodiazepines on NOS is related to the absence of substituents on N4 and to the absence of a halogen substituent on C5 phenyl group. Although benzodiazepine's inhibitions observed in this study are not in the physiological range in normal cases, these inhibitions could be significant in drug abuse situations and should be taken into account for the rational design of drugs which specifically inhibit NOS. PMID- 11731014 TI - Genes differentially expressed in the kindled mouse brain. AB - Kindling involves long-term changes in brain excitability and is considered a model of epilepsy and neuroplasticity. Differentially expressed genes in the kindled mouse brain were screened using an reverse transcription-polymerase chain reaction (RT-PCR) differential display (DD) method. C3H male mice were kindled with 40 stimuli in the hippocampus at 5-min intervals. Hippocampal RNA was isolated for DD from mice at 0.5 h, 1 day, 1 week, and 1 month after kindling and from sham-operated controls. About 30,000 bands were screened and of these, 50 were displayed differentially. Northern blot analysis confirmed that 26 of the 50 bands were differentially expressed following rapid kindling. Further sequence analysis revealed that 14 of the genes were previously identified and 12 were novel. The novel genes are referred to as King (1-12) genes because of their association with kindling. According to their temporal and quantitative pattern of expression in forebrain, the 26 genes were grouped into five types. Expression of five of the DD genes, one from each expression type, was further analyzed in hippocampus, forebrain, brainstem, and cerebellum of the kindled mice. Differential expression of these genes was observed in hippocampus and forebrain, but not in brainstem or cerebellum. Only one gene, a regulator of G-protein signaling 4 (RGS4), showed prolonged changes in expression in response to kindling. Our results show that rapid kindling produces spatial and temporal changes in gene expression that may influence kindling-associated neuroplasticity. PMID- 11731015 TI - Lactoferrin is synthesized by activated microglia in the human substantia nigra and its synthesis by the human microglial CHME cell line is upregulated by tumor necrosis factor alpha or 1-methyl-4-phenylpyridinium treatment. AB - The presence of the iron-binding protein lactoferrin (Lf) in some specific areas of the central nervous system and particularly in the normal human substantia nigra, where it is found in dopaminergic (DA) neurons and some glial cells, led us to investigate Lf synthesis in this area. Lf mRNA were identified using in situ hybridization and found in small ameboid cells. These cells were identified using immunocytochemistry as activated microglia since they exhibited macrophage markers such as the CD68 and the CR1 antigens. Double immunofluorescent labeling confirmed that the two Lf immunostained cell populations were activated microglia and DA neurons. Since activated microglia contained both Lf and its messenger, these cells are the Lf producing cells. The presence of Lf in DA neurons in which no Lf messengers were visible, might be due to an endocytosis mechanism, DA neurons probably internalizing Lf produced in microglial cells located in their neighborhood. In neuropathological disorders, such as Alzheimer's and Parkinson's diseases, inflammatory process and oxidative stress are events that contribute to neuronal death. Since Lf concentration increases during these pathologies, we studied the level of Lf expression under these different stresses and showed, using RT-PCR, that the immortalized human embryonic microglial CHME cell line produced Lf transcripts under tumor necrosis factor alpha or 1-methyl-4 phenylpyridinium treatment whereas untreated cells did not. These data confirm that Lf is produced only when microglia are activated. PMID- 11731017 TI - Expression of GABA(A) receptor subunits in the rat central nucleus of the inferior colliculus. AB - Expression of GABA(A) receptor (GABA(A)R) alpha(1), alpha(2), beta(2), gamma(1), gamma(2L) and gamma(2S) subunit mRNA was examined in three cell classes in the central nucleus of the rat inferior colliculus (CNIC). GABA(A)R alpha(1) and gamma(2L) subunit mRNA expression was greatest in large cells (over 25 microm long diameter), intermediate in medium sized cells (15 to 25 microm long diameter) and lowest in small cells (10 to 15 microm long diameter). GABA(A)R gamma(2S) and alpha(2) subunits had the opposite pattern, highest in the small cells, intermediate in medium cells and lowest in large cells. GABA(A)R beta(2) was significantly lower in small cells than the two other classes, while differences between large and medium cells were not significant. GABA(A)R gamma(1) subunit mRNAs expression was not above background in any of the three cell types assessed. The expression of GABA(A)R subunits suggests that cell classes in the rat CNIC may differ in their response to GABA and GABAergic drugs. PMID- 11731016 TI - Expression of catecholaminergic mRNAs in the hyperactive mouse mutant coloboma. AB - The SNAP-25 deficient mouse mutant coloboma (Cm/+) is an animal model for investigating the biochemical basis of locomotor hyperactivity. The spontaneous hyperactivity exhibited by coloboma is three times greater than control mice and is a direct result of the SNAP-25 deletion. SNAP-25 is a presynaptic protein that regulates exocytotic neurotransmitter release; coloboma mice express only 50% of normal protein concentrations. Previous research has determined that there is an increase in the concentration of norepinephrine but a decrease in dopamine utilization in the striatum and nucleus accumbens of coloboma mice. In situ hybridization analysis revealed that there were corresponding increases in tyrosine hydroxylase (TH) mRNA expression in noradrenergic cell bodies of the locus coeruleus of Cm/+ mice. In contrast, TH mRNA expression in substantia nigra appeared normal in the mutant mouse. alpha(2)-Adrenergic receptors are important modulators of central noradrenergic function and dopamine release. In situ hybridization data revealed that alpha(2A)-adrenergic receptor mRNA expression is upregulated in Cm/+ mice. These results suggest an underlying abnormality in noradrenergic regulation in this hyperactive mouse mutant. PMID- 11731018 TI - Ontogeny of AMPA receptor gene expression in the developing rat midbrain and striatum. AB - AMPA receptors mediate most of the fast excitatory synaptic transmission in the mammalian CNS. Their ontogeny during embryonic (E) and postnatal (P) development is still poorly understood. We have studied the expression of the genes encoding for AMPA glutamate receptor subunits (GlurA, GlurB, GlurC and GlurD) in the rat ventral mesencephalon (MES) and striatum (STR) and in fetal midbrain primary cultures. Each receptor subunit shows unique area- and temporal-expression pattern. In MES, GluRA, GlurB and GlurC mRNA are detectable from the earliest embryonic stage studied (E13) and raise thereafter between E15 and E17, to plateau at E19 to adult values. Differently, GlurD mRNA increases throughout embryonic and postnatal development reaching its highest levels in the adult MES. The pattern of AMPA proteins corresponded to the mRNA levels for all subunits. In the STR, GlurA gene expression increases between E15 and E19, GlurB mRNA levels are sustained from the first embryonic stages analyzed (E15) until E19 and gradually decrease thereafter toward adult levels, GlurC gene expression increases gradually throughout ontogeny to reach its highest levels in the adult. STR GlurD transcripts remain at constant levels in all stages studied. In embryonic MES primary cultures, every subunit show a characteristic expression profile similar to that observed in vivo. They all decrease significantly during the second week in vitro. Thus, all the AMPA receptor subunit transcripts appear independently regulated during development, probably depending on the tissue specific environment, which seems preserved in MES cultures. PMID- 11731019 TI - Divers pathways mediate delta-opioid receptor down regulation within the same cell. AB - Various mechanisms have been proposed for opioid receptor down regulation in different experimental preparations. The present study was aimed to test whether distinct mechanisms can mediate opioid receptor down regulation within the same cell. For this purpose we transfected HEK-293 cells with rat delta-opioid receptor (DOR). We exposed the cells to the opioid agonist etorphine in the absence or presence of various pharmacological agents and measured the binding of the opioid ligand [(3)H]diprenorphine to either isolated cell membranes or whole cells. We found that internalization of the receptors into the cell was mediated by clathrin coated pits and that the internalized receptors were degraded either in lysosomes or by proteosomes. Down regulation involved phosphorylation and at least two different kinases, a tyrosine kinase (TK) and MAPK kinase (MEK), mediated DOR down regulation in parallel routes. G-protein-coupled receptor kinase (GRK) was found to have only a minor role in DOR down regulation in HEK 293 cells. On the other hand, in N18TG2 cells that endogenously express delta opioid receptors, GRK was the predominant kinase mediating DOR down regulation, with only a minor role for TK and MEK. We conclude that down regulation can take place via divers pathways within the same cell, and that in different cells down regulation is mediated by different mechanisms, depending on the kinase profile of the cells and the compartmentalization of the receptors within the cells. PMID- 11731020 TI - Changes in voltage-gated calcium channel alpha(1) gene expression in rat dorsal root ganglia following peripheral nerve injury. AB - Although an increase in the excitability and ectopic spontaneous discharge (ESD) of primary sensory neurons can lead to abnormal burst activity, which is associated with neuropathic pain, the underlying molecular mechanisms are not fully understood. To investigate the relationship between these electrical abnormalities in injured neurons and voltage-gated calcium channel (VGCC) gene expression, reverse transcription-polymerase chain reaction (RT-PCR) was used to monitor the expression of the VGCC alpha(1) gene in the dorsal root ganglion (DRG) following chronic constriction injury (CCI) and axotomy of the rat sciatic nerve. Electrophoresis of the RT-PCR products showed the presence of multiple types of VGCC alpha(1) transcripts with various levels of basal expression in lumbar 4, 5, and 6 DRGs. CCI decreased alpha(1C), alpha(1D), alpha(1H), and alpha(1I) mRNA expression at 7 days in the ipsilateral DRG, to approximately 34 50% of the contralateral side. The same transcripts were repressed 7 days after sciatic axotomy and their reduction levels proved similar to those of CCI. Considering that changes of the intracellular calcium concentration modify the maintenance of ESD in injured DRG, these results suggest that the downregulation of alpha(1C), alpha(1D), alpha(1H) and alpha(1I) subunit gene expression in the rat DRG following peripheral nerve injury may contribute to the production of ESD associated with damaged nerves. PMID- 11731021 TI - Distinct phosphorylation patterns underlie Akt activation by different survival factors in neurons. AB - The survival of cultured cerebellar granule neurons can be maintained by depolarizing levels of potassium (high K(+), HK), insulin-like growth factor (IGF 1), cyclic AMP or lithium. We examined the possibility that the signaling pathways activated by these different factors converge and that Akt might represent such a point of convergence. Consistent with this possibility, we find that Akt is phosphorylated and activated by all four survival factors. The pattern of Akt phosphorylation induced by the four survival factors, however, shows differences. While IGF-1 induces phosphorylation of Akt at both Ser473 and Thr308, HK and cyclic AMP stimulate phosphorylation at Thr308 only. Lithium increases phosphorylation at Ser473 but not at Thr308. Our results are consistent with the possibility that Akt is a central component of different survival promoting pathways in granule neurons. The different phosphorylation patterns, however, point to a previously unappreciated complexity in the regulation of Akt activity in neurons. Finally, we provide evidence indicating that SGK, a kinase that is structurally related to Akt, is also activated by the four survival factors. PMID- 11731022 TI - Gene expression in the hippocampus of behaviorally stimulated rats: analysis by DNA microarray. AB - Gene expression in the hippocampus of rats exposed to a brief vigorous swimming task was analyzed by DNA microarray hybridization against 5184 known DNA sequences. Each of 12 rats consistently expressed 17% of the genes probed on the microarrays. Transcripts from genes for energy metabolism, development and differentiation, and gene regulation were expressed in the hippocampus to the highest degree relative to their availability on the microarray. The number of genes that were differentially expressed in behaviorally stimulated compared to undisturbed rats was estimated at 2.7% of all expressed genes. Behavioral effects were relatively greater on expression of genes for control of the cell cycle and apoptosis, development and differentiation, protein processing and gene regulation. The magnitude of differential expression due to behavioral stimulation was lower than that generally reported for developmental processes and disease states. Thus the impact of acute behavioral stimulation on steady state levels of gene expression was small, but (with four replicates per condition) detectable at a statistically significant level. The effects of behavioral stimulation on transcription may thus involve changes in the expression of a relatively small number of genes to a low but reproducible degree. PMID- 11731024 TI - Conditional gene targeting for cancer gene therapy. AB - Current treatment of solid tumors is limited by severe adverse effects, resulting in a narrow therapeutic index. Therefore, cancer gene therapy has emerged as a targeted approach that would significantly reduce undesired side effects in normal tissues. This approach requires a clear understanding of the molecular biology of both the malignant clone and the biological vectors that serve as vehicles to target cancer cells. In this review we discuss novel approaches for conditional gene expression in cancer cells. Targeting transgene expression to malignant tissues requires the use of specific regulatory elements including promoters based on tumor biology, tissue-specific promoters and inducible regulatory elements. We also discuss the regulation of both replication and transgene expression by conditionally-replicative viruses. These approaches have the potential to restrict the expression of transgenes exclusively to tissues of interest and thereby to increase the therapeutic index of future vectors for cancer gene therapy. PMID- 11731025 TI - Targeting gene expression using HSV vectors. AB - Herpes simplex virus (HSV) is an encapsulated DNA virus, with many favourable properties for use as a gene transfer vector. For gene therapy applications, it may be desirable to restrict transgene expression to pre-defined subsets of cells. One potential method for achieving targeted transgene expression using the HSV vector system might involve dictating the cell types to which the vector will transfer the therapeutic transgene of interest. HSV delivers its genetic payload to cells directly through the plasmalemma; the mechanisms are complex and involve multiple viral and cell surface determinants. We have investigated several ways in which each component of the cell entry cascade may be manipulated in order to restrict viral DNA and transgene delivery to particular cellular populations. Our results indicate that targeted transduction may be a viable approach to achieving our goal of targeted HSV-mediated transgene expression. PMID- 11731026 TI - Targeted drug conjugates: principles and progress. AB - Reports of targeting drugs using antibodies have appeared in the literature since 1958, but exciting clinical results in this field have only been reported in the last few years. Progress in this field has occurred largely through an understanding how drug-immunoconjugates work. The objective of this review is to draw together the fundamental principles on which this field of work is based, to examine the evidence supporting those principles, and the effectiveness and selectivity of targeted drug conjugates. The activity of many drug immunoconjugates can now largely be accounted for by the underlying principles. Excellent development work, both with conventional anti-cancer agents and very potent drugs have led to a number of interesting clinical trials. In the best Phase I and II trials, good evidence of effectiveness have been reported, which suggest that drug-immunoconjugates may now be heralding a new era for chemotherapy. PMID- 11731027 TI - Bioreductive and gene therapy approaches to hypoxic diseases. AB - Hypoxia is a feature that exists in most, if not all, solid tumours and hypoxia has been shown to exist in a variety of other diseases. Bioreductive prodrugs have been developed to preferentially target the hypoxic cells in tumours. They are prodrugs, that are reductively activated (catalysed by reductive enzymes) to afford their active (toxic) species. More recently, bioreductive delivery agents that "release" a therapeutic entity preferentially under hypoxic conditions have also been developed to target hypoxia, not only in tumours, but also in a host of other diseases. This new technology platform is described in this review. In addition, we discuss the potential of utilising hypoxia to deliver selective gene therapy based upon the transcription factor HIF-1 and the use of unique genetic sequences termed HRE's (hypoxia responsive elements) that specifically control gene expression under hypoxic conditions. Finally, we describe how these drugs and gene-based therapeutic approaches can be combined to potentially deliver a highly selective form of therapy for cancer and other diseases where hypoxia plays a major pathophysiological role. PMID- 11731028 TI - Drug targeting to hypoxic tissue using self-inactivating bioreductive delivery systems. AB - Hypoxia is a characteristic feature of a number of diseases including some cancers, rheumatoid arthritis and diabetes. Hypoxic tissue facilitates the use of bioreductive drug targeting systems as oxygen suppresses the release of the active drug. This review focuses on bioreductive delivery where accompanying intramolecular cyclisation negates adduct formation between the bioreductive and macromolecules such as DNA. To date, three systems have been reported. In the quinone lactonization system, reduction of the quinone facilitates through bond cyclisation and concomitant release of the drug. In the self-alkylating system, a nucleophile is built into the bioreductive structure to favour intramolecular cyclisation over nucleophilic attack from DNA moieties. The final system is based on vitamin E which undergoes redox mediated cyclisation between its oxidised (tocopherol quionone) and reduced (tocopherol) forms. Self-inactivating bioreductive delivery systems represent a powerful tool for extending bioreductive-based drug delivery to non-cancerous hypoxic tissues. PMID- 11731029 TI - Cyclodextrins in oligonucleotide delivery. AB - The aim of this contribution is to summarize recent findings on the potential use of cyclodextrins and their derivatives as carriers for oligonucleotide agents. Their peculiar properties could be exploited in such an emerging therapeutic area by virtue of their capability of interacting with cellular membranes, thus giving rise to improved cellular uptake. In particular, some specific derivatives could be considered as promising future excipients for the delivery of "naked" antisense and/or decoy oligonucleotides which are difficult to formulate with existing pharmaceutical excipients. PMID- 11731030 TI - Safety evaluation of a xylanase expressed in Bacillus subtilis. AB - A programme of studies was conducted to establish the safety of a xylanase expressed in a self-cloned strain of Bacillus subtilis to be used as a processing aid in the baking industry. To assess acute and subchronic oral toxicity, rat feeding studies were conducted. In addition, the potential of the enzyme to cause mutagenicity and chromosomal aberrations was assessed in microbial and tissue culture in vitro studies. Acute and subchronic oral toxicity was not detected at the highest dose recommended by OECD guidelines. There was no evidence of mutagenic potential or chromosomal aberrations. Furthermore, the organism used for production of the xylanase is already accepted as safe by several major national regulatory agencies. PMID- 11731031 TI - Studies on the inhibition of tumor cell growth and microtubule assembly by 3 hydroxy-4-[(E)-(2-furyl)methylidene]methyl-3-cyclopentene-1,2-dione, an intensively coloured Maillard reaction product. AB - Very recently, 3-hydroxy-4-[(E)-(2-furyl)methylidene]methyl-3-cyclopentene-1,2 dione (1) has been successfully identified as an intensively coloured Maillard product formed from glucose and L-proline upon thermal food processing. Using a biomimetic synthetic strategy, reference material of compound 1 was prepared and purified, and then used to study its effect on the growth of human tumor cells. Compound 1 was found to potently inhibit the growth of human tumor cells in vitro. Using a reporter gene assay we could show that in growth inhibitory concentrations compound 1 effectively inhibits the phosphorylation of the transcription factor Elk-1. In addition, 1 was found to affect the microtubule skeleton. The human mammary carcinoma cell line MCF-7 exhibits a decrease of the microtubule organisation when treated for 24 h with 1 (> or =20 microM). At concentrations of 30 microM and above a loss of microtubule integrity is observed after 1 h incubation. In vitro studies demonstrated that the polymerisation and, to a minor extent, also the depolymerisation of tubulin, isolated and purified from bovine brain, is inhibited in a dose-dependent manner at concentrations of 30 microM and above. This is the first time that a non-enzymatically formed browning compound of known structure was reported to effectively inhibit tumor cell growth and microtubule assembly. PMID- 11731032 TI - Strain-dependent in vitro and in vivo effects of oleic acid anilides on splenocytes and T cells in a murine model of the toxic oil syndrome. AB - The toxic oil syndrome is an exogenously-induced autoimmune disease in humans, which is believed to be due to the accidental ingestion of oleic acid anilides. In a previously established murine model anilides-treated A/J mice developed a wasting disease after 1 week. Anilides-treated B10.S mice showed after 6 weeks a hyperimmunglobulinemia with autoantibody production, but no clinical symptoms. We now compared in vitro the effects of anilides on splenocytes and T cells in A/J and B10.S mice. Splenocyte proliferation was similar in both strains. After in vivo treatment of mice with anilides and in vitro restimulation, splenocytes of sick A/J mice showed a significant increase in splenocyte proliferation. Splenocytes from B10.S mice, however, had a suppressed baseline response and did not proliferate on restimulation. Adherent cells were necessary to induce proliferation in A/J mice-derived T cells. Apoptosis in splenocytes was significantly elevated in anilides-treated A/J and in B10.S mice as compared to saline-treated controls. These data show that anilides are able to affect the immune system in a strain-dependent way and may therefore take part in inducing the disease seen in humans and mice. PMID- 11731033 TI - DNA damage in human fibroblasts exposed to fumonisin B(1). AB - Fumonisins are mycotoxins produced by several Fusarium species (Fusarium verticilloides and F. proliferatum) that infest corn and other cereals. Fumonisin B(1) (FB(1)), structurally resembling sphingoid bases, is an inhibitor of ceramide synthetase, a key enzyme involved in de novo sphingolipid biosynthesis and in the reacylation of free sphingoid bases derived from sphingolipid turnover. This inhibitory effect leads to accumulation of free sphinganine and sphingosine and subsequent induction of cell death. However, the downstream effectors activated by these sphingolipids in the cell death-signalling pathway are little known. The aim of this study was to evaluate, in FB(1)-exposed human fibroblasts, the involvement of oxygen free radicals and of some other biochemical pathways, caspase-3 activity, poly(ADP-ribose)polymerase (PARP) cleavage and DNA damage evaluated by comet assay. Our results indicate that FB(1) treatment (48, 72 h and 10, 50, 100 microM) does not affect cellular viability. Conversely, after 72 h of treatment, FB(1) (50 and 100 microM) induced DNA damage, an enhancement of caspase-3-activity and cleavage of PARP compared to controls. In addition, FB(1) increased the expression of HSP70 in a concentration and time-dependent manner. Our results indicate that DNA damage of apoptotic type in human fibroblasts is caused by exposure to FB(1) at high concentrations and for a prolonged time and that the genotoxic potential of FB(1) has probably been underestimated and should be reconsidered. PMID- 11731034 TI - Degradation of organochlorine pesticides by meat starter in liquid media and fermented sausage. AB - The effect of meat starter on the degradation of DDT and lindane was investigated. The insignificant role of Lactobacillus plantarum in degrading p,p' DDT and lindane presented in tryptone soya broth (TSB) and mineral salt medium (MSM) with or without 120 ppm nitrite was observed.The degradation of DDT and lindane by Micrococcus varians in TSB and MSM with or without nitrite were studied. The results indicated that DDT or lindane were degraded during the incubation period. The reduction in DDT at the end of the incubation period (15 days) was about 24.1 and 32.5% in TSB and MSM without nitrite, respectively. Corresponding values in the same media with nitrite were 37.5 and 46.4%. Regarding the reduction in lindane, it was recorded as 27.9 and 40.0% in TSB and MSM without nitrite, respectively and 38.4 and 48.4% in the same media with nitrite. The results indicated that culture media M. varians metabolized DDT mainly to DDD and lindane mainly to 2,4-, 2,5-, 2,6- and 3,4-dichlorophenol; 2,3,4- and 2,3,5-trichlorophenol; hexachlorobenzene; and pentachlorophenol. The effect of pesticides on the growth rate of meat starter was also investigated. The addition of DDT or lindane resulted in a slight decrease in counts of the strains during the initial incubation in TSB or MSM. Then the microorganisms recovered and began to grow logarithmically, but not as well as in a normal situation. The effect of fermentation stage by meat starter on DDT and lindane in fermented sausage was recorded. The results indicated that during the 72 h of fermentation, the reduction was 10 and 18% of DDT and lindane, respectively. These results confirmed that the fermentation process in meat products reduced pesticide residues and these reductions were due to the activity of meat starter. PMID- 11731035 TI - Lack of adverse effects in the F1 offspring maternally exposed to genistein at human intake dose level. AB - Recently there has been growing concern about endocrine disrupters (ED) derived from synthetic and natural chemicals. It has been argued that ED might cause developmental disorders in the next generations of animals and humans; however, this is still unclear. Therefore, we investigated whether maternal exposures to genistein (GEN) during gestation and lactation alter reproductive organs in the F1 offspring compared with those in 17beta-estradiol (E2)-maternally exposed F1 offspring. Pregnant Sprague-Dawley rats were treated orally with 0.4 mg/kg, 4.0 mg/kg GEN or 10 microg/kg E2. Maternal or neonatal effects on the number of live pups, implantation sites, sex ratio, anogenital distance, eyelid opening/vaginal opening and body weight of live pups were not altered by GEN or E2. The weights of reproductive organs at the adult stage F1 offspring were not altered by maternal exposure to GEN, except for the ventral prostate. However, the weight of the seminal vesicle was significantly decreased from postnatal day (PND) 21 to PND 70 in E2-treated offspring. Sperm analyses, cell count in seminiferous tubules and follicular development in the ovary were not altered by maternal exposure to GEN. Taken together, these results suggest that maternal exposure of GEN might not have adverse effects on the reproductive organs in the F1 offspring at the human intake dose level. PMID- 11731036 TI - Effect of dietary administration of genistein, nonylphenol or ethinyl estradiol on hepatic testosterone metabolism, cytochrome P-450 enzymes, and estrogen receptor alpha expression. AB - The objective of this study was to examine effects of estrogenic agents of varying potencies (genistein, p-nonylphenol, and ethinyl estradiol) on hepatic testosterone metabolism, cytochrome P-450 (CYP450) enzymes, and ERalpha expression. These endpoints were examined as potential biomarkers of, and contributors to, endocrine disruptive activity. Exposure occurred during critical developmental periods, from gestational day 7 through weaning via the mothers' diet. Thereafter, rats were exposed via their diet to the compounds until puberty (postnatal day 50). Testosterone hydroxylase and 5alpha-reductase activities, CYP2C and CYP3A levels were determined. In general, the compounds were more active in male rats than female rats. The only effect observed in female rats was at the 250 ppm genistein dose, in which an approximately 40% increase in 5alpha reductase activity was observed. In male rats, genistein treatment had mixed effects on testosterone metabolism. The 1250 ppm dose decreased both CYP2C and CYP3A protein levels. Nonylphenol had the most profound effects on testosterone metabolism and CYP450 expression in male rats, with effects occurring at doses as low as 25 ppm. An increase in 5alpha-reductase activity and a decrease in the formation of 16alpha-OH-, 2alpha-OH-testosterone metabolites, CYP2C and CYP3A protein were observed. EE2 decreased the formation of several testosterone metabolites and CYP2C protein. All compounds had some effect on hepatic ERalpha expression, although a consistent effect was not observed. This study demonstrates that the test compounds can influence hepatic testosterone hydroxylase activity and CYP450 expression, as well as ERalpha expression, although these activities cannot be directly related to estrogenic activity. PMID- 11731037 TI - Endocrine-disrupting effects of styrene oligomers that migrated from polystyrene containers into food. AB - The endocrine-disrupting effects of styrene dimers (SD: NSD-01, -08 and -09) and styrene trimers (ST: NST -01, -03 and -12), which migrated from polystyrene (PS) containers into instant food, were investigated together with styrene monomer (SM) using in vitro and in vivo assays. In the estrogen (ER) and androgen receptor (AR) binding assay, SM, SD and ST showed no binding activity at concentration of 10(-10)-10(-5) mol/l. In order to evaluate the estrogenic activity in vivo, the uterotrophic assay was conducted. When prepubertal and ovariectomized adult rats were dosed with SM, SD and ST for 3 days by subcutaneous injection, these compounds did not induce significant increase in uterine weight. Additionally, to evaluate anti-androgen activity in vivo, the Hershberger assay for anti-androgenic activity in the presence of testosterone treatment was conducted. When castrated, testosterone-treated immature male rats were dosed SM, SD and ST for 7 days by oral gavage, these compounds did not induce a decrease in the seminal vesicle, ventral prostate and levator ani plus bulbocavernosus muscle weights. To evaluate the effects on hormones other than sex hormones, the thyroid hormone receptor (TR) binding assay and rat serum prolactin (PRL) was conducted. In the TR binding assay, SM, SD and ST showed no binding activity at a concentration of 10(-5) mol/l. When ovariectomized rats were dosed with SM, SD and ST for 3 days by sc injection, the results showed there was no change in rat serum PRL. From the above these results, we concluded that SM, SD and ST exhibit no apparent estrogenic, androgenic, anti-androgenic and thyroid activity. PMID- 11731038 TI - Evaluation of the potential effects of ingredients added to cigarettes. Part 1: cigarette design, testing approach, and review of results. AB - A testing program was designed to evaluate the potential effects of 333 ingredients added to typical commercial blended test cigarettes on selected biological and chemical endpoints. Ingredients were incorporated into the test cigarettes as they are normally used in the manufacturing process. The studies performed included a bacterial mutagenicity screen (Ames assay), a mammalian cell cytotoxicity assay (neutral red uptake), determination of smoke chemical constituents, and a 90-day nose-only smoke inhalation study in rats. Three pairs of test cigarettes were produced, each containing one of three different groups of ingredients. In each pair, one of the cigarettes contained the normal approximate use level of the ingredients (low-level) and the other a 1.5-3 multiple of the normal use level (high-level). Analysis of the test cigarettes for selected ingredients or markers indicated that the target application rates were achieved and that the cigarettes had been manufactured as intended. Evaluation of cigarette performance indicated that the addition of the ingredients at high levels did not significantly alter the burning characteristics of the test cigarettes. Specific details of the individual studies conducted as part of an ingredient evaluation program are discussed in Parts 2-4 of this publication series (Food and Chemical Toxicology, 2002, 40, 93 104; Food and Chemical Toxicology, 2002, 40, 105-111; Food and Chemical Toxicology, 2002, 40, 113-131). The results of the smoke chemistry studies indicated a reduction in the majority of the smoke constituents and a few isolated instances of increases when compared to the control cigarettes. These smoke chemistry changes, while statistically significant, were not supported by any significant alteration in the biological effects of cigarette smoke normally seen with the bacterial mutagenicity assay, cytotoxicity assay or subchronic inhalation study. Based on the results of these studies, it can be concluded that these ingredients added to tobacco do not add significantly to the overall toxicity of cigarettes. PMID- 11731039 TI - Evaluation of the potential effects of ingredients added to cigarettes. Part 2: chemical composition of mainstream smoke. AB - Cigarette mainstream smoke from blended research cigarettes with and without the addition of ingredients was analyzed for its chemical composition. In total, 333 ingredients commonly used in cigarette manufacturing were assigned to three different groups. Each group of ingredients was introduced at a low and a high level to the test cigarettes. The list of the 51 smoke constituents determined is based on those analytes suggested for analysis in a US Consumer Product Safety Commission proposal for low ignition cigarettes and cigarette smoke constituents identified by the International Agency for Research on Cancer as worthy of concern and characterized as carcinogens. An increase in the yield of total particulate matter (TPM) in the range of 13 to 28% relative to the control cigarette without ingredients was observed for all test cigarettes. This was presumably caused by the higher transfer rates of the added ingredients to the smoke compared to the transfer from the tobacco part of the filler. When the yields of individual constituents were normalized to the TPM yields, a reduction in the majority of the constituents was observed when compared to the control. For one of the ingredient groups this reduction was especially high: for phenols a maximum of 70%, for polycyclic aromatic hydrocarbons 50%, and for N nitrosamines 45%. An increase in the amount relative to TPM was observed for a few smoke constituents: hydrogen cyanide and cadmium (one ingredient group), formaldehyde (one ingredient group), and resorcinol and lead (two ingredient groups). These results are consistent with the lack of any increased activity in the in vitro and in vivo assays in this same series of studies (Food and Chemical Toxicology 2002, 40, 105-111; Food and Chemical Toxicology 2002, 40, 113-131). An overall assessment of our data suggests that these ingredients, when added to the tobacco, do not add to the toxicity of smoke, even at the elevated levels tested in this series of studies. PMID- 11731040 TI - Evaluation of the potential effects of ingredients added to cigarettes. Part 3: in vitro genotoxicity and cytotoxicity. AB - Cigarette mainstream smoke from blended cigarettes with and without the addition of ingredients was assayed for its cytotoxicity and genotoxicity. In total, 333 ingredients commonly used in cigarette manufacturing were assigned to three different groups. Each group of ingredients was added at a low and a high level to the test cigarettes. The mutagenicity of the particulate phase of the resulting cigarette smoke was assayed in the Salmonella plate incorporation (Ames) assay with tester strains TA98, TA100, TA102, TA1535 and TA1537. The cytotoxicity of the gas/vapor phase and the particulate phase was determined in the neutral red uptake assay with mouse embryo BALB/c 3T3 cells. Within the sensitivity and specificity of the test systems, the in vitro mutagenicity and cytotoxicity of the cigarette smoke were not increased by the addition of the ingredients. PMID- 11731041 TI - Evaluation of the potential effects of ingredients added to cigarettes. Part 4: subchronic inhalation toxicity. AB - Mainstream smoke from blended research cigarettes with (test) and without (control) the addition of ingredients to the tobacco was assayed for inhalation toxicity. In total, 333 ingredients commonly used in cigarette manufacturing were assigned to three different groups. Each group of ingredients was introduced at a low and a high level to the test cigarettes. Male and female Sprague-Dawley rats were exposed nose-only either to fresh air (sham) or diluted mainstream smoke from the test, the control, or the Reference Cigarette 1R4F at a concentration of 150 microg total particulate matter/l for 90 days, 6h/day, 7 days/week. A 42-day post-inhalation period was included to evaluate reversibility of possible findings. There were no remarkable differences in in-life observations or gross pathology between test and control groups. An increase in activity of liver enzymes, known to be due to the high smoke dose, revealed no toxicologically relevant differences between the test and control groups. No toxicological differences were seen between the test and control groups for smoke-related hematological changes, such as a decrease in total leukocyte count. The basic smoke-related histopathological effects, which were more pronounced in the upper respiratory tract than in the lower respiratory tract, were hyperplasia and squamous metaplasia of the respiratory epithelium, squamous metaplasia and atrophy of the olfactory epithelium, and accumulation of pigmented alveolar macrophages. There were no relevant qualitative or quantitative differences in findings in the respiratory tract of the rats exposed to the smoke from the control and test cigarettes. The data indicate that the addition of these 333 commonly used ingredients, added to cigarettes in three groups, did not increase the inhalation toxicity of the smoke, even at the exaggerated levels used. PMID- 11731043 TI - Doppler quantitative measures by region to discriminate prostate cancer. AB - This study was conducted to assess if sonographic discrimination between healthy and cancerous prostate tissue might be improved using regional analysis of ultrasound (US) Doppler measures. A total of 39 subjects underwent 3-D Doppler sonography before radical prostatectomy. Cancer locations were identified from hematoxylin and eosin (H&E)-stained slides. Three prostate data volumes consisting of a frequency shift and power-mode Doppler US and whole mount histology images were spatially registered for each prostate, then divided into entirely 1 mL-sized regions of cancerous or noncancerous tissue. Each prostate was visually divided into a peripheral and a periurethral region within which US Doppler measures were calculated. Receiver operating characteristic (ROC) and simulated biopsy analyses within each prostate were performed. Mean speed in colored pixels (V), and speed-weighted pixel density (SWD) are good discriminators for prostate cancer in the periurethral and the peripheral regions, respectively. Using SWD in a simulated biopsy yields increased cancer detection in the peripheral region. PMID- 11731044 TI - Real-time spatial compound imaging improves reproducibility in the evaluation of atherosclerotic carotid plaques. AB - Compound imaging has the ability of reducing speckle and clutter artifacts demonstrated in in vitro studies compared to conventional, single-angle imaging. We investigated intra- and interobserver agreement of 38 outlines of carotid artery plaque images acquired by these techniques, by measuring the overlapping area after repeated outlines. In general, both techniques showed good agreement. When considering the images with poorest overlap, compound imaging had a significant advantage over conventional imaging regarding both intra- and interobserver agreement. The interobserver variation for the overlapping area after two outlines was 20% for conventional technique and 10% for compound. The interobserver variation of the gray scale median value (GSM) for conventional technique ranged from -32 to +20 and from -6 to +6 for compound. Likewise, the coefficient of repeatability for the GSM value was 13 for conventional imaging and three for compound imaging, and interobserver variation for the GSM value for the overlapping area was 34% and 9% for conventional and compound technique. In conclusion, compound imaging improves intra- and interobserver agreement and reduces interobserver variation in the GSM value in a clinical setting. PMID- 11731045 TI - Assessing spectral algorithms to predict atherosclerotic plaque composition with normalized and raw intravascular ultrasound data. AB - Spectral analysis of backscattered intravascular ultrasound (IVUS) data has demonstrated the ability to characterize plaque. We compared the ability of spectral parameters (e.g., slope, midband fit and y-intercept), computed via classic Fourier transform (CPSD), Welch power spectrum (WPSD) and autoregressive (MPSD) models, to classify plaque composition. Data were collected ex vivo from 32 human left anterior descending coronary arteries. Regions-of-interest (ROIs), selected from histology, comprised 64 collagen-rich, 24 fibrolipidic, 23 calcified and 37 calcified-necrotic regions. A novel quantitative method was used to correlate IVUS data with corresponding histologic sections. Periodograms of IVUS samples, identified for each ROI, were used to calculate spectral parameters. Statistical classification trees (CT) were computed with 75% of the data for plaque characterization. The remaining data were used to assess the accuracy of the CTs. The overall accuracies for normalized spectra with CPSD, WPSD and MPSD were, respectively, 84.7%, 85.6% and 81.1% (training data) and 54.1%, 64.9% and 37.8% (test data). These numbers were improved to 89.2%, 91.9% and 89.2% (training) and 62.2%, 73% and 59.5% (test) when the calcified and calcified-necrotic regions were combined for analysis. Most CTs misclassified a few fibrolipidic regions as collagen, which is histologically acceptable, and the unnormalized and normalized spectra results were similar. PMID- 11731046 TI - Does B-mode common carotid artery intima-media thickness differ from M-model? AB - An increased intima-media thickness of the common carotid artery is thought to be an early sign of atherosclerosis. Both B- and M-mode ultrasonographic techniques are used to measure the intima-media thickness of the common carotid artery (B IMT and M-IMT, respectively). The present study compares intima-media thickness of the common carotid artery measured with the two techniques. Intima-media thickness was measured in a random population sample of 250 subjects. Comparison was made by mean and 95% confidence intervals of differences between B-IMT and M IMT, by linear regression analysis, and by intraclass and concordance correlation coefficients. M-IMT was + 0.011 +/- 0.091 mm (95% confidence intervals: -0.167 to + 0.188 mm) larger than B-IMT, which was 0.661 +/- 0.136 mm (range: 0.380 to 1.120 mm). Intraclass and concordance correlation coefficients were 0.802 and 0.801, respectively. In conclusion, acceptable agreement exists between the two methods and there was no important systematic difference between B-IMT and M-IMT. PMID- 11731047 TI - Assessment of bone status using speed of sound at multiple anatomical sites. AB - Studies in vitro and in vivo have shown that quantitative ultrasound (QUS) is a valid tool for the assessment of bone status. Current QUS methods using the transmission technique are limited to one peripheral bone site. A new system, Sunlight Omnisense (Omnisense, Sunlight Medical Ltd., Rehovot, Israel), measures speed of sound (SOS, in m/s) along the surface of the bone based on an axial transmission technique. The Omnisense can measure SOS at several anatomical sites. This study evaluated the SOS at different anatomical sites in a healthy population. A total of 334 adult women from three research centers in the USA and Canada with a mean (+/- SD) age of 48.8 (+/- 17.4) years were enrolled in this study. SOS was measured at the proximal third phalanx, distal one third radius, midshaft tibia, and fifth metatarsal. The mean SOS (+/- SD) values for the phalanx, radius, tibia and metatarsal were 3984 (+/- 221), 4087 (+/- 147), 3893 (+/- 150) and 3690 (+/- 246) m/s, respectively. Each anatomical site SOS was significantly different (p < 0.001) from that of the other sites. SOS at the different anatomical sites was modestly, but significantly, correlated (r = 0.31 to 0.56, p < 0.001). Similar correlation coefficients were obtained for the T scores. The mean T scores for subjects over the age of 60 years were -1.94, 2.01, -0.97 and -1.42 for the phalanx, radius, tibia and metatarsal, respectively. The age of peak SOS and the rate of change thereafter varied with anatomical site, implying that the prevalence of osteopenia and osteoporosis was site-dependent if only one T score cut-off point was used. Comparing individuals, 10% to 17% of patients had T scores that differed by more than a factor of 2 between sites. Weight and age were some of the contributing factors to this heterogeneity. The Omnisense provides an opportunity to assess bone status at different anatomical sites. Whether or not combining measurements from all these anatomical sites will improve osteoporosis management still needs to be determined. PMID- 11731048 TI - A freehand elastographic imaging approach for clinical breast imaging: system development and performance evaluation. AB - A prototype freehand elastographic imaging system has been developed for clinical breast imaging. The system consists of a fast data acquisition system, which is able to capture sequences of intermediate frequency echo frames at full frame rate from a commercial ultrasound scanner whilst the breast is deformed using hand-induced transducer motion. Two-dimensional echo tracking was used in combination with global distortion compensation and multi-compression averaging to minimise decorrelation noise incurred when stress is applied using hand induced transducer motion. Experiments were conducted on gelatine phantoms to evaluate the quality of elastograms produced using the prototype system relative to those produced using mechanically induced transducer motion. The strain sensitivity and contrast-to-noise ratio of freehand elastograms compared favourably with elastograms produced using mechanically induced transducer motion. However, better dynamic range and signal-to-noise ratio was achieved when elastograms were created using mechanically induced transducer motion. Despite the loss in performance incurred when stress is applied using hand-induced transducer motion, it was concluded that the prototype system performed sufficiently well to warrant clinical evaluation. PMID- 11731049 TI - Experimental imaging of the acoustic nonlinearity parameter B/A for biological tissues via a parametric array. AB - The acoustic nonlinearity parameter B/A is an important parameter in nonlinear acoustics and has been suggested to be a novel parameter for ultrasound tissue characterization. In this paper, the principle and experimental results for B/A imaging by using a parametric array are presented. As two primary waves with different frequencies are radiated simultaneously from a circular piston source, a secondary wave at the difference frequency is generated due to the nonlinear interaction of the two primary waves. The axial sound pressure amplitude curve for the generated difference frequency wave in the near field is measured and agrees well with the theoretical calculation. Unlike the second harmonic wave, the difference frequency component of the parametric array grows almost linearly with distance from the piston source and goes through fewer oscillations in the near field. It therefore provides a better source for acoustic nonlinearity parameter tomography. On the basis of the finite amplitude insert substitution method and the conventional CT technology, an experimental imaging system for acoustic nonlinearity parameter B/A via a parametric array is set up. B/A images for several biological specimens, including normal and pathological tissues, are obtained. PMID- 11731050 TI - Contrast agent stability: a continuous B-mode imaging approach. AB - The stability of contrast agents in suspensions with various dissolved gas levels has not been reported in the literature. An in vitro investigation has been carried out that studied the combined effect of varying the acoustic pressure along with degassing the suspension environment. In this study, the contrast agents were introduced into suspensions with different oxygen concentration levels, and their relative performance was assessed in terms of decay rate of their backscatter echoes. The partial pressures of oxygen in those solutions ranged between 1.5 and 26 kPa. Two IV and one arterial contrast agents were used: Definity, Quantison, and Myomap. It was found that Quantison and Myomap released free bubbles at high acoustic pressure that also dissolved faster in degassed suspensions. The backscatter decay for Definity did not depend on the air content of the suspensions. The destruction of bubbles was dependent on acoustic pressure. Different backscatter performance was observed by different populations of bubbles of the last two agents. The physical quantity of "overall backscatter" (OB) was defined as the integral of the decay rate over time of the backscatter of the contrast suspensions, and improved significantly the understanding of the behaviour of the agents. A quantitative analysis of the backscatter properties of contrast agents using a continuous imaging approach was difficult to achieve. This is due to the fact that the backscatter in the field of view is representative of a bubble population affected by the ultrasound (US) field, but this bubble population is not representative of the contrast suspension in the whole tank. Single frame insonation is suggested to avoid the effects of decay due to the ultrasonic field, and to measure a tank-representative backscatter. The definition of OB was useful, however, in understanding the behaviour of the agents. PMID- 11731051 TI - Quantification of ultrasound contrast agent response: comparison of continuous wave Doppler and power Doppler to backscattered radiofrequency data. AB - Our goal was to compare two quantification methods of ultrasound contrast agents available in clinical practice [continuous wave Doppler intensity (CWDI) and power Doppler intensity (PWDI)] to the reference technique (radio-frequency analysis) with a simple recirculating flow phantom using a renal dialysis cartridge. Measurements were made at different doses of perflenapent emulsion and BR1. Cineloops of power Doppler images were recorded using a clinically available ultrasound unit (HDI 3000). Simultaneously, integrated backscatter (IBS) was measured by analysis of radiofrequency signals, whereas Doppler signal intensity was measured with a continuous wave Doppler device. A linear relationship was found between CWDI and IBS and between PWDI and IBS when R(2) was calculated for each pair of parameters injection-by-injection. Results are summarized by the average R(2) for all injections between CWDI and IBS (BR1: R(2) = 0.93 +/- 0.05, perflenapent emulsion: R(2) = 0.94 +/- 0.03) and between PWDI and IBS (BR1: R(2) = 0.88 +/- 0.07, perflenapent emulsion: R(2) = 0.79 +/- 0.09). However, for all data obtained from all different injected doses and for both contrast agents, there was considerable variation of CWDI and PWDI values measured for a given value of IBS. In conclusion, for a fixed microbubble population, CWDI and PWDI can be proposed for quantification of USCA. However, their important variations observed at each dose make it difficult to link a single value of PWDI or CWDI or IBS to a single microbubble distribution composition. PMID- 11731052 TI - Poroelastography: imaging the poroelastic properties of tissues. AB - In the field of elastography, biological tissues are conveniently assumed to be purely elastic solids. However, several tissues, including brain, cartilage and edematous soft tissues, have long been known to be poroelastic. The objective of this study is to show the feasibility of imaging the poroelastic properties of tissue-like materials. A poroelastic material is a material saturated with fluid that flows relative to a deforming solid matrix. In this paper, we describe a method for estimating the poroelastic attributes of tissues. It has been analytically shown that during stress relaxation of a poroelastic material (i.e., sustained application of a constant applied strain over time), the lateral-to axial strain ratio decreases exponentially with time toward the Poisson's ratio of the solid matrix. The time constant of this variation depends on the elastic modulus of the solid matrix, its permeability and its dimension along the direction of fluid flow. Recently, we described an elastographic method that can be used to map axial and lateral tissue strains. In this study, we use the same method in a stress relaxation case to measure the time-dependent lateral-to-axial strain ratio in poroelastic materials. The resulting time-sequenced images (poroelastograms) depict the spatial distribution of the fluid within the solid at each time instant, and help to differentiate poroelastic materials of distinct Poisson's ratios and permeabilities of the solid matrix. Results are shown from finite-element simulations. PMID- 11731053 TI - Measurements of bubble-enhanced heating from focused, MHz-frequency ultrasound in a tissue-mimicking material. AB - Time-resolved measurements of the temperature field in an agar-based tissue mimicking phantom insonated with a large aperture 1-MHz focused acoustic transducer are reported. The acoustic pressure amplitude and insonation duration were varied. Above a critical threshold acoustic pressure, a large increase in the temperature rise during insonation was observed. Evidence for the hypothesis that cavitation bubble activity in the focal zone is the cause of enhanced heating is presented and discussed. Mechanisms for bubble-assisted heating are presented and modeled, and quantitative estimates for the thermal power generated by viscous dissipation and bubble acoustic radiation are given. PMID- 11731054 TI - Continuous and pulsed ultrasound do not increase heat shock protein 72 content. AB - Therapeutic ultrasound (US) is a common treatment used in the rehabilitation of injured muscle. To determine whether therapeutic US could increase the content of heat shock protein (HSP) 72 in skeletal muscle, male Sprague-Dawley rats were anesthetized and the muscles from one hind limb treated with 15 min of US at 1 MHz using either: 1. continuous US at 1.0 W/cm(2), 2. pulsed US at 2.0 W/cm(2) at 50% duty cycle, or 3. pulsed US at 1.0 W/cm(2) at 20% duty cycle. All treatments were applied using a transducer (1.6-cm diameter) on an area of the rat hind limb twice the size of the sound head. At 24 h following treatment, the plantaris, soleus, white and red gastrocnemius muscles were removed and assessed for HSP 72 content by Western blotting. No significant increases in HSP 72 content were detected in any of the muscles examined following any US treatment. These results suggest muscle HSP content is not elevated following a typical therapeutic dose of either continuous or pulsed US in the rat. PMID- 11731055 TI - Accuracy of an electromagnetic three-dimensional ultrasound system for carotid artery imaging. AB - Freehand, three-dimensional (3-D) ultrasound (US) systems, which incorporate an electromagnetic tracking device to register the 3-D spatial location of images acquired using a standard linear array scan-probe, are a flexible and cost effective solution for many clinical applications. The reconstruction accuracy of one such system was investigated by using a precision-made phantom. The error in 3-D distance measurements, under conditions appropriate to US investigations of the carotid arteries, was found to be -0.45 +/- 1.30 mm, equivalent to -0.53 +/- 3.39% (mean +/- SD). The results are relevant to data acquired using a single sweep scan and for distances in the range 25.00 to 79.06 mm. Both the overall accuracy and precision in point-target location were found to be relatively unaffected by scan depth, and the precision of point-target location was found to be poorest in the elevational direction. In conclusion, the system tested in our laboratory performed with high accuracy, adopting a setup and scan-sweep identical to that used for imaging of the carotid arteries in 3-D. PMID- 11731056 TI - Thermal effects of focused ultrasound energy on bone tissue. AB - The effects of focused ultrasound (US) at therapeutic acoustic power levels were studied in vivo on the bone-muscle interface in rabbit thighs. The purpose of this study was to provide direction in establishing safety guidelines for treating tissue masses using focused US on or near bone. A positioning device was used to manipulate a focused US transducer (1.5 MHz) in a magnetic resonance imaging (MRI) scanner. This system was used to sonicate the femurs of 10 rabbits at acoustic power levels of 26, 39, 52 and 65 W for 10 s. The rabbits were euthanized either 4 h or 28 days after the sonications and the bone samples were harvested for histology examinations. In the femurs studied, acoustic power levels from 39 to 65 W resulted in soft tissue damage characterized grossly by coagulated tissue and bone damage depicted by yellow discoloration. Histologic examination of lesions from sonications from 39 to 65 W demonstrated that osteocyte damage and necrosis, characterized by pyknotic cells and empty lacunae, occurred within the ablation area extending through the bone. The follow-up MR images demonstrated an increase in the amount of damage in the femurs at 28 days posttreatment in comparison to images taken immediately after treatment. Focused US directed at the femur caused immediate significant thermal damage to bone in the form of osteocyte necrosis extending through the (approximately) 1 cm bone in this study. The results suggest that, when focused US energy is directed at or near bone-muscle interfaces, precautions should be taken to avoid thermal damage to the bone that can compromise its strength for extended periods. PMID- 11731057 TI - "Stable" inertial cavitation. AB - This note compares theoretical predictions of pressure waves scattered by free gas bubbles with recent acoustical determinations of cavitation thresholds for individual microbubbles of the surfactant-stabilized contrast agent Sonazoid(R). The results indicate that surfactant-coated microbubbles undergo "stable" (i.e., repetitive) inertial cavitation above a threshold of 0.3 to 0.4 MPa at 2.5 MHz, and that irreversible postcollapse bubble fragmentation usually requires much higher pressures (approximately 1.5 MPa). Adverse bioeffects can be expected in vivo far below these fragmentation pressures when contrast agents are present. With diagnostically relevant exposures, the threshold for the generation of petechiae in skeletal muscle is approximately 0.6 MPa at 2.5 MHz. PMID- 11731059 TI - Venipuncture-induced neuropathic pain: the clinical syndrome, with comparisons to experimental nerve injury models. PMID- 11731060 TI - A new model of sciatic inflammatory neuritis (SIN): induction of unilateral and bilateral mechanical allodynia following acute unilateral peri-sciatic immune activation in rats. AB - Immune activation near healthy peripheral nerves may have a greater role in creating pathological pain than previously recognized. We have developed a new model of sciatic inflammatory neuritis to assess how such immune activation may influence somatosensory processing. The present series of experiments reveal that zymosan (yeast cell walls) acutely injected around the sciatic nerve of awake unrestrained rats rapidly (within 3h) produces low threshold mechanical allodynia in the absence of thermal hyperalgesia. Low (4 microg) doses of zymosan produce both territorial and extra-territorial allodynia restricted to the ipsilateral hindpaw. Higher (40-400 microg) doses of zymosan again produce both territorial and extra-territorial allodynia. However, allodynia is now expressed both in the ipsilateral as well as contralateral hindpaws. Several lines of evidence are provided that the appearance of this contralateral ('mirror') allodynia reflects local actions of zymosan on the sciatic nerve rather than spread of this immune activator to the general circulation. Since many clinical neuropathies result from inflammation/infection of peripheral nerves rather than frank physical trauma, understanding how immune activation alters pain processing may suggest novel approaches to pain control. PMID- 11731061 TI - Reduced development of tolerance to the analgesic effects of morphine and clonidine in PKC gamma mutant mice. AB - A variety of second messenger systems have been implicated in the intracellular mechanisms of tolerance development to the analgesic actions of morphine, a mu opioid, and clonidine, an alpha-2 adrenergic receptor agonist. Here, we studied mice that carry a null mutation in the gene encoding a neuronal specific isoform of protein kinase C (PKC), namely, PKC gamma. We used the tail-flick test to construct dose-response curves before and 4 days after chronic morphine (75-mg pellets, subcutaneously (s.c.)) or clonidine treatment (0.3mg/kg, s.c., twice daily). Baseline tail-flick latencies did not differ in PKC gamma mutant and wild type mice (3-4s). Both morphine and clonidine produced a dose-dependent suppression of the tail-flick response with an ED(50) (effective dose resulting in a 50% reduction of the control response) value (2.0mg/kg for morphine and 0.1mg/kg for clonidine) that was similar for naive mutant and wild-type mice. In contrast, after 4 days of drug delivery, mutant mice showed significantly less rightward shift in the dose-response curve to morphine (six-fold for wild-type and three-fold for mutant mice) and to clonidine (five-fold for wild-type and no shift for the mutant mice). These results indicate that PKC gamma contributes to the development of tolerance to the analgesic effects of both morphine and clonidine. Chronic morphine treatment can also result in sensitization of spinal cord neurons and increased pain behaviors following a noxious insult. To assess the contribution of PKC gamma to this process, we studied the responses of wild type and mutant mice to an intraplantar injection of formalin (a model of persistent pain) following chronic morphine treatment. Although morphine tolerance increased formalin-evoked persistent pain behavior and Fos-LI in wild type mice, there was no difference between placebo- and morphine-treated mutant mice, suggesting that PKC gamma also contributes to chronic morphine-induced changes in nociceptive processing. PMID- 11731062 TI - Botulinum toxin A for the treatment of chronic neck pain. AB - A clinical study tested the therapeutic efficacy of Botulinum toxin A (BTXA) when injected into symptomatic neck muscles after one injection session. Patients with chronic neck pain were randomly assigned to receive either a high dose of an active treatment or an injection of the same volume of normal saline. Patients were compared for 4 months using a comprehensive set of outcome measures that included the Neck Pain and Disability Scale (Spine 24 (1999) 1290) and pressure algometry (Arch Phys Med Rehabil 67 (1986) 406; Pain 30 (1987) 115; Clin J Pain 2 (1987) 207). Analyses were consistent in showing significant benefits from the injection session; however, the effects were not specific to the group treated with BTXA. Both treatment and control groups showed a significant decline in pain and disability across time and an increased ability to withstand pressure on trigger points. The heavy incidence of adverse events in the treatment group may partly explain the absence of a treatment effect specific to BTXA. The results show that a single dose treatment without physical therapy is not effective for chronic neck pain. PMID- 11731063 TI - Extending the Aberdeen Back Pain Scale to include the whole spine: a set of outcome measures for the neck, upper and lower back. AB - Many therapists treat the spine as a 'functional unit', but suitable condition specific outcome measures for the whole spine are not available. One of the most rigorously tested measures for back pain related health status is the Aberdeen Back Pain Scale, but it is only suitable for the lower back. The Aberdeen Back Pain Scale was extended to create a set of interlocking outcome measures for the neck, upper and lower back. Questions in these extended outcome measures had to fulfil a series of psychometric criteria before being accepted into the final questionnaires. This involved response frequency, item-total correlation and principal component analysis. The final questionnaires were tested for reliability, criterion and construct validity, responsiveness and acceptability, on patients attending a primary care musculoskeletal clinic. One question was discarded from all three sets of questionnaires and a further question was removed from the neck questionnaire. Baseline scores approximated to normal distributions. Although not completely reproducible, they were internally consistent, so showed evidence of reliability. They were highly correlated with the SF-12 and their mean scores changed according to whether their health status, measured by a transition question, improved, stayed the same or worsened. Modified standardised response means showed large changes when health status improved and moderate-sized changes when health status worsened. Patients made few adverse comments about the questionnaires and found them acceptable. The Extended Aberdeen Spine Pain Scales for neck, upper and lower back pain, showed evidence of reliability, validity, responsiveness and acceptability. They can be used for single regions of the spine or combined as clinically necessary. They are particularly recommended for primary care patients. PMID- 11731064 TI - Intra-articular morphine as analgesic in temporomandibular joint arthralgia/osteoarthritis. AB - The aim of this study was to determine the analgesic efficacy of a single dose intra-articular injection (i.a.) of morphine in 53 patients with unilateral arthralgia/osteoarthritis of the temporomandibular joint (TMJ). This randomized, double-blind, parallel group, multicenter study included a screening visit, a treatment visit, and a follow-up visit 1 week after treatment. Recordings of visual analog scales (VAS) pain intensity scores at maximum mouth opening (main efficacy variable) and at jaw rest were made directly before a 1-ml i.a. injection into one TMJ of either 1.0mg morphine-HCl, 0.1mg morphine-HCl, or saline (placebo). The pain intensity was also recorded at the follow-up and in a diary 3 days before and 5 days after the injection. The VAS pain score at maximum mouth opening was considerably reduced 1-10h after injection but without significant differences between groups. At the follow-up, the median VAS pain score at maximal mouth opening was significantly lower in the 0.1-mg morphine group than in the 1.0-mg morphine group (P<0.043) or the saline group (P<0.021). A significant increase in pain pressure threshold over the affected joint was seen in the 0.1-mg morphine group compared with the saline group at the follow-up but not 1 and 2h post-injection. The incidence of adverse events was small and did not differ between the treatment groups. In conclusion, one i.a. injection of 0.1mg morphine significantly increased the pain pressure threshold and mouth opening ability, but evidence for the analgesic property of the locally applied opioid was inconclusive. No dose-effect relation and no significant short-term analgesic property were seen. Although statistically significant, the magnitude of the reduced VAS pain intensity score was not clinically relevant at the 1-week follow-up. PMID- 11731065 TI - Vasodilatation in hyperalgesic rat skin evoked by stimulation of afferent A beta fibers: further evidence for a role of dorsal root reflexes in allodynia. AB - In areas of secondary hyperalgesia, innocuous mechanical stimuli evoke pain (allodynia). We have proposed that this is produced by a central pre-synaptic interaction whereby A beta-fibers evoke spike activity (dorsal root reflexes) in nociceptive afferents (Pain, 68 (1996) 13). This activity should conduct centrally, evoking allodynia, and peripherally, evoking neurogenic vasodilatation. Here we tested this hypothesis by examining the effects of electrical stimulation of A beta-fibers on cutaneous blood flow before and after producing secondary hyperalgesia in anesthetized rats. Cutaneous blood flow was recorded in the hind paw skin innervated by the sural nerve using a laser Doppler flowmeter. The sural nerve was prepared for electrical stimulation, and the evoked activity was recorded from the sciatic nerve in continuity. Electrical stimulation (1 Hz, 4 x 0.2 ms pulses, 20 s) was applied to the sural nerve at 2T (A beta-fibers only) and 4T and 6T (A beta + A delta-fibers). Flux was recorded at baseline and after capsaicin or mustard oil application outside the sural nerve territory. The effects of intravenous administration of the calcitonin gene related peptide (CGRP) receptor antagonist, alpha-CGRP(8-37), or of section of the sciatic nerve or of the L4-L6 dorsal roots were examined. Selective activation of the sural nerve A beta-fibers reliably evoked increases in cutaneous blood flow close to areas of chemical irritation or skin damage. A beta fiber-evoked vasodilatation was abolished by sciatic nerve or dorsal root section and had a spatial arrangement and optimal stimulation pattern suggesting a central synaptic interaction similar to that responsible for dorsal root reflexes. The flux increases were dose-dependently and reversibly inhibited by alpha-CGRP(8-37), indicating that the A beta-fiber-evoked vasodilatation resulted from the antidromic activation of nociceptive cutaneous afferent fibers. These results support our hypothesis by showing activation of nociceptive primary afferents by A beta-fibers in areas of allodynia in a manner consistent with a pre-synaptic interaction evoking dorsal root reflexes. PMID- 11731066 TI - A painful peripheral neuropathy in the rat produced by the chemotherapeutic drug, paclitaxel. AB - Paclitaxel, an effective anti-neoplastic agent in the treatment of solid tumors, produces a dose-limiting painful peripheral neuropathy in a clinically significant number of cancer patients. Prior work has demonstrated paclitaxel induced neurodegeneration and sensory loss in laboratory rodents. We describe here an experimental paclitaxel-induced painful peripheral neuropathy. Adult male rats were given four intraperitoneal injections on alternate days of vehicle or 0.5, 1.0, or 2.0 mg/kg of paclitaxel (Taxol). Behavioral tests for pain using mechanical and thermal stimuli applied to the tail and hind paws, and tests for motor performance, were taken before, during and after dosing for 22-35 days. All three doses of paclitaxel caused heat-hyperalgesia, mechano-allodynia, mechano hyperalgesia, and cold-allodynia, but had no effect on motor performance. Neuropathic pain began within days and lasted for several weeks. We did not detect any dose-response relationship. Tests at the distal, mid, and proximal tail failed to show evidence of a length-dependent neuropathy. Vehicle control injections had no effect on any measure. No significant systemic toxicities were noted in the paclitaxel-treated animals. Light-microscopic inspection of the sciatic nerve (mid-thigh level), L4-L5 dorsal root ganglia, and dorsal and ventral roots, and the gray and white matter of the L4-L5 spinal cord, showed no structural abnormalities. Electron microscopic examination of the sciatic nerve (mid-thigh level) and the L4-L5 dorsal root ganglia and dorsal horns demonstrated no degeneration of myelinated and unmyelinated axons in the sciatic nerve and roots, but revealed endoneurial edema. This model may be useful in understanding a significant source of pain in cancer patients, and in finding ways to avoid the neurotoxicity that limits paclitaxel therapy. PMID- 11731067 TI - Conscious and automatic uses of memory in chronic pain patients. AB - Memory deficits in chronic pain patients are frequently observed. The objective of this study was to explore memory performances of chronic pain patients by using the Process Dissociation Procedure developed by Jacoby (J. Mem. Lang. 30 (1991) 513). This procedure permits to separate the contribution of controlled processes from automatic processes operating within a memory task. The results show a significant decrease of controlled processes in chronic pain patients. Furthermore for both groups, automatic processes contribute in a similar extent to the memory performance. The estimates of controlled processes in the chronic pain patients are significantly related to the fear of pain and catastrophic beliefs. This is interpreted as a sign of interference between the attention consumed by pain experience (namely fear related to pain) and the attention to be allocated to the memory task. PMID- 11731068 TI - Adenosine inhibits excitatory transmission to substantia gelatinosa neurons of the adult rat spinal cord through the activation of presynaptic A(1) adenosine receptor. AB - Although intrathecal administration of adenosine analogues or A(1) adenosine receptor agonists is known to result in antinociception, this has not been examined yet at the cellular level. In the present study, we examined in pharmacology an action of adenosine on glutamatergic miniature excitatory postsynaptic currents (mEPSCs) in substantia gelatinosa (SG) neurons of an adult rat spinal cord slice; this was done under the condition where a postsynaptic action of adenosine was blocked. In 65% of the neurons examined (n=72), adenosine at a concentration of 100 microM depressed the frequency of mEPSC in a reversible manner; the remaining neurons exhibited an inhibition followed by potentiation of the frequency. When examined quantitatively in extent in some cells (n=25), the inhibition was 40+/-3% (n=25) while the potentiation was 42+/-8% (n=6). These actions were not accompanied by a change in mEPSC amplitude. The inhibitory action on mEPSC frequency was dose-dependent in a range of 10-500 microM with an EC(50) value of 277 microM. The inhibitory action of adenosine was mimicked by a selective A(1) adenosine receptor agonist, CPA (1 microM; depression: 54+/-9%, n=4); this action of adenosine (100 microM) was not observed in the presence of a specific A(1) adenosine receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) (1 microM; 94+/-4% of control, n=3). The facilitatory action of adenosine (100 microM) was unaffected by an A(2a) antagonist, ZM 241385 (0.1 microM, n=3); an A(2a) agonist, CGS 21680 (0.1-10 microM; n=6), was without actions on mEPSC frequency. It is concluded that adenosine inhibits excitatory transmission to SG neurons through the activation of presynaptic A(1) adenosine receptor and that some of the inhibition is followed by a potentiation of the transmission. It remains to be examined which subtypes of adenosine receptors except for the A(1)- and A(2a)-subtypes are involved in the potentiating action. Considering that adenosine-like immunoreactivity and adenosine receptors are expressed at a high density in the SG, which is thought to play an important role in modulating nociceptive transmission from the periphery to the central nervous system, this inhibitory action of adenosine could contribute to a negative modulation of pain transmission. PMID- 11731070 TI - Neosporosis in bovine dairy herds from the west of France: detection of Neospora caninum DNA in aborted fetuses, seroepidemiology of N. caninum in cattle and dogs. AB - Neospora caninum is considered one of the major causes of abortion in cattle in most parts of the world. In this study, the role of N. caninum was investigated in groups of aborted cattle and dairy herds from the west of France. Good correlation was found between parasite DNA detection in fetuses and serologic statuses of dams. In groups with documented abortion status and no antibodies to other pathogens, 17-45% of aborted animals were seropositive for N. caninum, and significant relationship between prevalence of Neospora antibodies and frequency of abortions was found. Neospora-associated abortions were observed all the year round, with a peak in summer. Higher ratios of seropositive abortions were found before the 6th month of gestation. In 12 herds studied in the field, serologic prevalence ranged 6-47%. No difference in age was found between seropositive and seronegative cows. Results indicate that N. caninum is an important and stable cause of abortion in cattle in France. PMID- 11731071 TI - Bovine neosporosis: immunoblot improves foetal serology. AB - The purpose of the study is the comparative evaluation of the immunofluorescent antibody test (IFAT) and an immunoblot (IB) test for the examination of foetal fluids for specific antibodies against Neospora caninum. Peritoneal and pleural fluids as well as abomasal contents were analysed. The results of the serological examinations were compared to those obtained by histological, immunohistochemical, and PCR analysis of foetal tissues as well as to the results of maternal serological examinations. Fluids were used undiluted in the IB and reactions against six immunodominant antigens were recorded. When the recognition of at least two immunodominant antigens was regarded as positive, the agreement of the IB with other diagnostic methods was good to moderate as characterised by kappa-values of 0.76 (histology/immunohistochemistry), 0.69 (maternal serology) and 0.54 (PCR on foetal tissues). The IB results agreed better with the results of the other diagnostic methods than those of the IFAT. The higher relative sensitivity of the IB was regarded as the main reason for the better agreement. However, also the specificity of the IB was superior to that of the IFAT in relation to histology/immunohistochemistry, maternal serology and PCR. PMID- 11731072 TI - Evaluation and improvement of parasitological tests for Trypanosoma evansi infection. AB - Research was undertaken to critically evaluate parasitological tests for the detection of Trypanosoma evansi in blood. The relative sensitivity of mouse inoculation (MI), the haematocrit centrifugation technique (HCT) and a modified miniature anion-exchange centrifugation technique (MAECT) were compared using blood and buffy coat. The effect that storage of blood prior to inoculation into mice has on the reliability of the MI test was also evaluated. The tests may be ranked in increasing order of sensitivity: HCT, MAECT with whole blood, MI with whole blood, MAECT with buffy coat and MI with buffy coat. The latter was able to detect 1.25 T. evansi per 4ml of blood. The reliability of the MI test was not reduced with storage of blood containing at least 25 T. evansi per ml for up to 21h prior to inoculation into mice. These results demonstrate that sensitivity of the MI and MAECT are increased approximately 10-fold through the use of buffy coat in place of whole blood. Although, the MI is marginally more sensitive MAECT is better suited to field use. PMID- 11731073 TI - The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: towards the development of a fungal controlled release device. AB - Studies showed that chlamydospores of the nematophagous fungus, Duddingtonia flagrans, are capable of surviving pressures of several tonnes when incorporated into matrices and pressed into tablets for the manufacture of prototype intraruminal controlled release devices (CRDs). They remain viable in this tabletted form for at least 9 months when stored at 4 degrees C. In vitro studies demonstrated that there was no effect on spore viability of prolonged exposure to either room or elevated temperature (40 degrees C) in air, or under an atmosphere of either of the major ruminal gases, carbon dioxide and methane. In vivo, studies showed that viable chlamydospores could be detected at the erosion surface of prototype CRDs recovered from the rumen and also in faeces of fistulated sheep, for up to 3 weeks after administration. Further studies have shown that chlamydospores released from such devices can substantially reduce the number of infective larvae that develop in cultures of faeces collected from sheep infected with the nematode parasite, Haemonchus contortus. This work demonstrates, in principle, that the deployment of chlamydospores of D. flagrans in intraruminal CRDs, is another possibility in the development of a range of methods for the biological control of parasites in livestock. PMID- 11731075 TI - The potential of nematophagous fungi to control the free-living stages of nematode parasites of sheep: feeding and block studies with Duddingtonia flagrans. AB - A series of feeding trials was conducted with penned sheep harboring Trichostrongylus colubriformis infections. They were offered barley grains supporting the growth of the nematophagous fungus Duddingtonia flagrans. It was shown that as little as 5g of grain/sheep per day was sufficient to virtually eliminate larval numbers from faecal culture. This effect persisted for the time that the fungal grains were fed, and for up to 2 days following cessation of feeding this material. Macerated fungal grains were also incorporated into a range of feed block formulations. In all these, D. flagrans was found to survive the manufacturing process and resulted in significant reductions in larval numbers in faecal cultures set up during the feeding period to sheep. This was observed even for sheep that showed only modest and irregular block consumption. These studies demonstrate that supplementary feeding or block administration offer potential deployment options for D. flagrans as a means of biological control of nematode parasites of livestock. PMID- 11731074 TI - Morphologic and genetic characterization of Onchocerca lupi infecting dogs. AB - In the past decades, sporadic cases of ocular Onchocerca infection have been reported in canids in US and Europe. The present study was undertaken to provide a detailed description of the morphologic characteristics of adults and microfilariae and to characterize the 5S ribosomal rRNA gene (5S rDNA) spacer sequences of Onchocerca lupi causing canine onchocercosis. The morphology of O. lupi is unique within the genus, and morphology based cluster analysis indicates that O. lupi is not closely related to the members of domestic cattle or horse clades occurring in North America and Europe. Similarly, the signature of the 5S rDNA spacer sequences of O. lupi does not resemble any other Onchocerca 5S rDNA spacer sequences including those of the members of domestic cattle or horse clades. The adult and microfilarial morphology and sequence signature supports the biological arguments that a distinct species, O. lupi and not O. lienalis, is responsible for canine ocular onchocercosis. PMID- 11731076 TI - Effect of 0.29% w/w fipronil spray on adult flea mortality and egg production of three different cat flea, Ctenocephalides felis (Bouche), strains infesting cats. AB - To evaluate the effect of fipronil spray on adult flea mortality and flea egg production of three different cat flea, Ctenocephalides felis (Bouche) strains, 30 domestic short hair cats were randomly allocated into six groups of five cats each. On day 0, cats in groups 2, 4 and 6 were treated with fipronil at 5-6ml/kg. Cats in groups 1, 3 and 5 served as untreated controls. On days -2, 7, 14, 21, and 28 each cat was infested with 50 adult cat fleas. Groups 1 and 2 were infested with fleas from the Kansas1 Colony (KS1) strain. Groups 3 and 4 were infested with a recently colonized cat flea strain from Florida (R6). Groups 5 and 6 were infested with fleas from the ARC strain. The adulticidal activity of fipronil was determined by flea comb counts 48h after treatment and then 48h after each reinfestation. Any flea eggs produced during the infestations were collected and counted prior to the 48h comb counts. Fipronil spray was > or = 99.5% effective against adults of all three cat flea strains when applied during an active infestation. Fipronil spray provided > or = 98.2 and > or = 99.5% control of adult fleas and egg production, respectively, for all strains through week 2. On days 23 and 30 control of R6 adults and egg production was significantly lower than either the ARC or the KS1 strain. On day 30, control of R6 adults and egg production was 77.3 and 87.3%, respectively. Control of KS1 adults and egg production on day 30 was significantly lower than the ARC strain. Fipronil provided > or = 99.5 and > or = 99.9% control of ARC fleas and egg production, respectively, throughout the entire study. The susceptibility to fipronil for the three strains was also evaluated on filter paper pesticide bioassays. The R6 strain was found to be less susceptible than the KS1 and ARC strains. The LC(95) estimates for the strains were 10.13, 4.77 and 2.62mg/m(2) for the R6, ARC and KS1 strains, respectively. PMID- 11731077 TI - Identification of screwworms, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), with a monoclonal antibody-based enzyme-linked immunosorbent assay (MAb-ELISA). AB - Myiasis caused by screwworms, Cochliomyia hominivorax (Coquerel), is devastating to warm-blooded animals and economically important to livestock producers. It is difficult to distinguish these pests, immature screwworms, from immatures of other non-pest fly species that often occur in animal wounds; it would be helpful to have tools available that do not rely on morphological characteristics. We developed two monoclonal antibodies (MAbs), highly specific for the screwworm, and used them in an enzyme-linked immunosorbent assay (MAb-ELISA), that differentiated screwworm eggs, larvae, pupae, and adults from those of the closely related secondary screwworm, C. macellaria (Fabricius) as well as Phormia regina (Meigen), Phaenicia sericata (Meigen), Calliphora vicina Robineau Desvoidy, and Chrysomya rufifacies (Macquart). In a blind study, the microplate MAb-ELISA, which took about 4h to complete, displayed high specificity (99%), sensitivity (92%) and overall accuracy (97%) in distinguishing all life stages of the screwworm. Electrophoresis results suggested that the two monoclonal antibodies recognized identical conformational epitopes present in all screwworm life stages. The screwworm eradication program, successful in eradicating this pest from the US, Mexico, most of Central America and Libya (after an accidental introduction), could benefit in future eradication, surveillance, and exclusion efforts by developing a reliable field identification kit based on MAb-ELISA that accurately and quickly distinguished cases of screwworm myiasis. PMID- 11731078 TI - A new type of class I bacterial 5-enopyruvylshikimate-3-phosphate synthase mutants with enhanced tolerance to glyphosate. AB - Glyphosate or Roundup is the most extensively used herbicide for broad-spectrum control of weeds. Glyphosate inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants. Applying the staggered extension process, we randomly mutated and recombined the aroA genes of Salmonella typhimurium and Escherichia coli to obtain four variants that exhibit significantly enhanced tolerance to glyphosate. All four mutants are chimeras of the two parental genes and, in addition, three of them carry one or more de novo point mutations. None of the amino acid substitutions in the mutants was in a position previously known to be important for catalysis or substrate binding. Kinetic analysis of EPSPS activity from these mutants indicated that the tolerance was attributed to a 2-10-fold increased specific activity, 0.4-8-fold reduced affinity to glyphosate, and 2.5 19-fold decreased K(m) for phosphoenolpyruvate. Such mutants will be instrumental for the structural and function study of the enzyme and for the generation of transgenic crops resistant to the herbicide. PMID- 11731079 TI - High expression of 92 kDa type IV collagenase (matrix metalloproteinase-9) in canine mammary adenocarcinoma. AB - The 92 kDa type VI collagenase (matrix metalloproteinase-9 (MMP-9)) activities on zymography assay were found to be 1-6 times higher in benign tumor breast tissues of 12 canines and 4-26 times higher in adenocarcinoma breast tissues of nine canines than that of control tissues, respectively. A full-length canine MMP-9 cDNA was cloned from the adenocarcinoma tissue by reverse transcription-PCR and 5'- and 3'-RACE. The isolated cDNA contained an open reading frame coding for a polypeptide of 704 amino acids. The predicted protein sequence displayed extensive similarity to that of known MMP-9s and contained a putative signal sequence, a propeptide, an active site with three zinc-binding histidine residues, a calcium-binding domain, a hemopexin region, and three key cysteine residues. Western blotting using MMP-9-specific antibodies prepared against the peptide corresponding to Arg(642)-Asp(704) of canine MMP-9 and Northern blotting using a MMP-9-specific cDNA fragment as a probe confirmed that MMP-9 (the 92 kDa protein band) was highly expressed in canine mammary adenocarcinoma tissues. Higher levels of MMP-9 activity were found in the sera of canines with mammary adenocarcinoma. The results indicated that MMP-9 plays an important role in the progression of a canine mammary tumor and that assay of serum MMP-9 is helpful for early diagnosis as progress of adenocarcinoma. PMID- 11731080 TI - Efficient gene transfer by fiber-mutant adenoviral vectors containing RGD peptide. AB - One of the hurdles to adenovirus (Ad)-mediated gene transfer is that Ad vectors mediate inefficient gene transfer into cells lacking in the primary receptors, Coxsackievirus and adenovirus receptor (CAR). We previously developed a fiber mutant Ad vector containing the Arg-Gly-Asp (RGD)-containing peptide motif on the HI loop of the fiber knob, and showed that the mutant vector had enhanced gene transfer activity to human glioma cells, which showed little CAR expression, compared to the vector containing wild type fiber. In this study, the feasibility of the Ad vector containing RGD peptide on the fiber knob was examined in a wide variety of cell types: CAR-positive or -negative human tumor cells, mouse cells, and leukemia cells. The mutant vector infected the cells, which lacked CAR expression but showed alpha(v) integrin expression, about 10-1000 times more efficiently than the vector containing wild type fiber via an RGD-integrin (alpha(v)beta3 and alpha(v)beta5)-dependent, CAR-independent cell entry pathway. The results of this study indicate that Ad vector containing RGD peptide on the fiber knob could be of great utility for gene therapy and gene transfer experiments. PMID- 11731081 TI - Characteristics of adenovirus-mediated tetracycline-controllable expression system. AB - The combination of recombinant adenovirus (Ad) vectors and the tetracycline controllable expression system is clearly an advantage in gene therapy and gene transfer experiment. In this study, we examined the characteristics of Ad vectors containing the tet-off or tet-on system. The Ad vector containing the tet-off system showed tightly regulatable transgene expression even at low MOI (multiplicity of infection). In contrast, regulation of gene expression by the Ad vector containing the tet-on system was not tight at low MOI, while it showed moderate regulation at high MOI (MOI=100). The Ad vector-mediated tet-on system showed lower inducible and higher background (basal) luciferase production than that of the Ad vector-mediated tet-off system. Moreover, the former system required a concentration of doxycycline, a derivative of tetracycline, approx. 2 3 log orders higher than that of the latter system to switch the luciferase expression. A combination of the vector containing the tet-on system and the vector containing the tetracycline-controlled transcriptional silencer (tTS) gene reduced the background luciferase production and improved regulation. These results suggest that the Ad vector containing the tet-off system is considered to be functionally superior to the vector containing the tet-on system. Care should be taken regarding regulation (especially lower inducibility and higher background), which is decreased in the Ad vector-mediated tet-on system in comparison with the tet-off system. The Ad vector containing the tetracycline controllable expression system should offer a powerful tool for gene therapy and gene transfer experimentation. PMID- 11731082 TI - Oxidative modification of human ceruloplasmin by peroxyl radicals. AB - Ceruloplasmin (CP), the blue oxidase present in all vertebrates, is the major copper-containing protein of plasma. We investigated oxidative modification of human CP by peroxyl radicals generated in a solution containing 2,2'-azobis(2 amidinopropane) dihydrochloride (AAPH). When CP was incubated with AAPH, the aggregation of proteins was increased in a time- and dose-dependent manner. Incubation of CP with AAPH resulted in a loss of ferroxidase activity. Superoxide dismutase and catalase did not protect the aggregation of CP, whereas hydroxyl radical scavengers such as ethanol and mannitol protected the protein aggregation. The aggregation of proteins was significantly inhibited by the copper chelators, diethyldithiocarbamate and penicillamine. Exposure of CP to AAPH led to the release of copper ions from the enzyme and the generation of protein carbonyl derivatives. Subsequently, when the amino acid composition of CP reacted with AAPH was analyzed, cysteine, tryptophan, methionine, histidine, tyrosine, and lysine residues were particularly sensitive. PMID- 11731083 TI - Purification and characterization of a galactose-specific lectin from corn (Zea mays) coleoptile. AB - We purified and characterized a lectin from the corn coleoptyle (Zea mays). The lectin (CCL) was purified by affinity chromatography on a Lactosyl-Sepharose 4B column. It is a glycoprotein of 88.7 kDa, composed mainly by glutamic, aspartic, glycine, and Ser residues; in a minor proportion, it contained methionine and cysteine residues. Carbohydrates that constituted 12% of the total weight comprised galactose, mannose, and N-acetyl-D-glucosamine. The lectin contained the blocked amino-terminus. Analysis of the lectin, determined from peptides obtained after trypsin digestion by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight), indicated that CCL has 18% homology with a putative calcium-dependent Ser/Thr protein kinase, from Arabidopsis thaliana, and 39% homology with a NADPH-dependent reductase from Z. mays. The lectin showed hemagglutinating activity toward several erythrocytes, including human A, B, and O. Hapten inhibition assays indicated that the lectin interacts specifically with the OH on C4 from galactose residues. OH- on C1 plays a relevant role in the interaction with CCL, since beta-galactose residues are better recognized than those from the anomeric alpha-galactose. Lack of lectin activity was observed in corn extracts; the highest specific activity was obtained from coleoptyle obtained at the 7th day after seeding. PMID- 11731084 TI - Inhibition and regulation of rat liver L-threonine dehydrogenase by different fatty acids and their derivatives. AB - Rat liver L-threonine dehydrogenase is a mitochondrial enzyme which transforms L threonine either into aminoacetone or into acetyl-CoA. We show that it is inhibited by several fatty acids and their derivatives: short chain fatty acids, L-2-hydroxybutyrate and D-3-hydroxybutyrate, long chain fatty acids, such as lauric acid, myristic acid, palmitic and stearic acids, bicarboxylic acids such as malonic acid and its derivatives methyl- and hydroxymalonic acids. The inhibition occurs at low and physiological concentrations of such compounds, which are normally present and metabolized in mitochondria. It presumably plays a role in the physiology of acetyl-CoA-dependent formation of fatty acids and ketobodies, in L-threonine-dependent gluconeogenesis, and in the regulation of L threonine metabolism by L-threonine dehydrogenase and L-threonine deaminase. PMID- 11731085 TI - The enhancement of electrostriction caused by lowering the solvent dielectric constant leads to the decrease of activation energy in trypsin catalysis. AB - The contribution of electrostriction of the solvent to the stabilization of the negatively charged tetrahedral transition state of a trypsin-catalyzed reaction was probed by means of kinetic studies involving high-pressure and solvent dielectric constant. A good correlation was observed between the increased catalytic efficiency of trypsin and the decreased solvent dielectric constant. When the dielectric constant of the solvents was lowered by 4.68 units, the loss of activation energy and that of free energy of activation were 2.26 kJ/mol and 3.09 kJ/mol, respectively. The activation volume for k(cat) decreased significantly as the dielectric constant of the solvent decreased, indicating that the degree of electrostriction of the solvent around the charged tetrahedral transition state has been enhanced. These observations demonstrate that the increase in the catalytic efficiency of the trypsin reaction with decreasing dielectric constant resulted from the stabilization of electrostatic energy for the formation of an oxyanion hole, and this stabilization was caused by the increase of electrostricted water around the charged tetrahedral transition state. Therefore, we conclude that control of the solvent dielectric constant can stabilize the tetrahedral transition state, and this lowers the activation energy. PMID- 11731086 TI - Effect of thymosin peptides on the chick chorioallantoic membrane angiogenesis model. AB - The effect of alpha- and beta-thymosin peptides, namely prothymosin alpha (ProT(alpha)), thymosin alpha(1) (T(alpha)1), parathymosin alpha (ParaT(alpha)), thymosin beta(4) (Tbeta4), thymosin beta(10) (Tbeta10), and thymosin beta(9) (Tbeta9), on the angiogenesis process was investigated using the chick chorioallantoic membrane as an in vivo angiogenesis model. The thymosin peptides tested were applied in 10 microl aliquots containing 0.01-4 nmoles of Tbeta4, Tbeta10 or Tbeta9, 0.016-6.66 nmoles of T(alpha)1, 4.1 pmoles-1.66 nmoles of ProT(alpha), and 4.4 pmoles-1.76 nmoles of ParaT(alpha). Phorbol 12-myristate 13 acetate and hydrocortisone were also used as positive and negative control, respectively. Tbeta4, ProT(alpha) and T(alpha)1 were found to enhance angiogenesis, while Tbeta10, Tbeta9 and ParaT(alpha) exhibited an inhibitory effect on the angiogenesis process. When mixtures of Tbeta4 and Tbeta10 containing active amounts of the two peptides at different proportions were applied, the promoting effect of Tbeta4 on angiogenesis was reversed in the presence of increasing concentrations of Tbeta10 and vice versa. The effect of Tbeta10, Tbeta9, ProT(alpha) and ParaT(alpha), in parallel with Tbeta4 and T(alpha)1, on the angiogenesis process was investigated for the first time as far as we know and the results of this study offer more insight into the biological regulatory roles of thymosin peptides, and provide helpful information about their therapeutic potential. Whether these agents could be used either as inhibitors of angiogenesis in disease states where uncontrolled angiogenesis is involved, e.g. in carcinogenesis, or as angiogenesis promoters that could be useful in wound healing, fracture repair, peptic ulcers etc., remains to be further studied. PMID- 11731087 TI - Differential regulation of gamma-glutamyltransferase mRNAs in four human tumour cell lines. AB - Human gamma-glutamyltransferase (GGT) belongs to a multigenic family and at least three mRNAs are transcribed from the gene that codes for an active enzyme. Four human tumour cell lines (HepG2, LNCap, HeLa and U937) with different GGT levels were used to investigate how GGT activity, total GGT mRNA and each individual GGT mRNA subtype responded to tumour necrosis factor-alpha (TNF-alpha), 12-O tetradecanoylphorbol 13-acetate (TPA) or sodium butyrate treatment. Butyrate reduced the GGT activity in HepG2 cells, and the level of total GGT mRNA accordingly, whereas TNF-alpha and TPA did not alter these parameters. In LNCap cells, TNF-alpha, TPA, and butyrate reduced the activity as well as the level of GGT total mRNA. In HeLa cells no significant changes were observed either in activity or in mRNA level whereas TPA induced both GGT activity and mRNA levels in U937 cells. The distribution of each GGT mRNA subtype (A, B and C) was found to be cell specific: type B mRNA was the major form in HepG2 cells, while type A was the major form in LNCap and HeLa, type A and type C were expressed almost at the same level in U937 cells. The GGT mRNA subtypes were also differently modulated in these cells after TNF-alpha, TPA or butyrate treatment, suggesting that they are regulated by distinct and cell type specific mechanisms. PMID- 11731088 TI - Inhibitory effects of phloroglucinol derivatives from Mallotus japonicus on nitric oxide production by a murine macrophage-like cell line, RAW 264.7, activated by lipopolysaccharide and interferon-gamma. AB - An aqueous acetone extract of the pericarps of Mallotus japonicus (MJE) inhibited nitric oxide (NO) production by a murine macrophage-like cell line, RAW 264.7, which was activated by lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). Seven phloroglucinol derivatives isolated from MJE exhibited inhibitory activity against NO production. Among these phloroglucinol derivatives, isomallotochromanol exhibited strong inhibitory activity toward NO production, exhibiting an IC(50) of 10.7 microM. MJE and the phloroglucinol derivatives significantly reduced both the induction of inducible nitric oxide synthase (iNOS) protein and iNOS mRNA expression. NO production by macrophages preactivated with LPS and IFN-gamma for 16 h was also inhibited by MJE and the phloroglucinol derivatives. Furthermore, MJE and the derivatives directly affected the conversion of L-[(14)C]arginine to L-[(14)C]citrulline by the cell extract. These results suggest that MJE and the phloroglucinol derivatives have the pharmacological ability to suppress NO production by activated macrophages. They inhibited NO production by two mechanisms: reduction of iNOS protein induction and inhibition of enzyme activity. PMID- 11731089 TI - Dioxygenative cleavage of C-methylated hydroquinones and 2,6-dichlorohydroquinone by Pseudomonas sp. HH35. AB - The dioxygenolytic catabolism of five C-methylated hydroquinones and 2,6 dichlorohydroquinone in Pseudomonas sp. strain HH35 was elucidated. This organism, which is known to catabolise 2,6-dimethylhydroquinone by 1,2-cleavage, accumulated metabolites from 2-methyl-, 2,3-dimethyl-, 2,5-dimethyl-, 2,3,5 trimethyl- and 2,3,5,6-tetramethylhydroquinone which we isolated and characterised by mass spectrometry and (1)H NMR and UV spectroscopy. The identification of these metabolites defined the impact of methyl groups present in the hydroquinone and showed how each substitution pattern determined the site of the initial enzymic attack. With the exception of the 2,3,5,6 tetramethylhydroquinone, all C-methylated hydroquinones were catabolised by an initial dioxygenolytic cleavage occurring adjacent (1,2- or 3,4-cleavage) to a hydroxy group. In addition, our results indicated that the 2,6 dichlorohydroquinone is catabolised in a similar way by this strain. PMID- 11731090 TI - Factor X activator from Vipera lebetina snake venom, molecular characterization and substrate specificity. AB - Our studies of the venom from the Levantine viper Vipera lebetina have demonstrated the existence of both coagulants and anticoagulants of the hemostatic system in the same venom. We showed that V. lebetina venom contains factor X activator (VLFXA) and factor V activator, fibrinolytic enzymes. VLFXA was separated by gel filtration on Sephadex G-100 superfine and ion exchange chromatography on CM-cellulose and on TSK-DEAE (for HPLC) columns. VLFXA is a glycoprotein composed of a heavy chain (57.5 kDa) and two light chains (17.4 kDa and 14.5 kDa) linked by disulfide bonds. VLFXA has multiple molecular forms distinguished by their isoelectric points. The differences in their pI values may be caused by dissimilarities in the respective charged carbohydrate content or in the primary sequence of amino acids. We synthesized 6-9 amino acid residues containing peptides according to physiological cleavage regions of human factor X and human factor IX. The peptides (Asn-Asn-Leu-Thr-Arg-Ile-Val-Gly-Gly - factor X fragment, and Asn-Asp-Phe-Thr-Arg-Val-Val-Gly-Gly - factor IX fragment) were used as substrates for direct assay of VLFXA. Cleavage products of peptide hydrolysis and the molecular masses of cleavage products of human factor X were determined by MALDI-TOF MS. The MALDI-TOF MS was highly efficient for the recovery and identification of peptides released by VLFXA hydrolysis. We can conclude that VLFXA cleaves the Arg(52)-Ile(53) bond in the heavy chain of human factor X and the Arg(226)-Val(227) bond in human factor IX precursor. VLFXA could not activate prothrombin nor had any effect on fibrinogen, and it had no arginine esterase activity toward benzoylarginine ethyl ester. PMID- 11731091 TI - Acetylphenylhydrazine induced haemoglobin oxidation in erythrocytes studied by Mossbauer spectroscopy. AB - The oxidative action of acetylphenylhydrazine (APH) on red blood cells obtained from healthy donors and from patients with breast cancer has been investigated by Mossbauer spectroscopy. Whole blood was incubated with APH for different time periods and the Mossbauer spectra of the packed red cells were recorded and compared. The evolution with time of the oxidation products has been followed. The largest difference in red cells analysis between healthy persons and patients was found after about 50 min of treatment where Mossbauer spectra of patient samples show a much broader spectral pattern due to an advanced haemoglobin oxidation. PMID- 11731092 TI - Identification of a cDNA clone encoding for a galactose-binding lectin from peanut (Arachis hypogaea) seedling roots. AB - A cDNA clone obtained from developing peanut (Arachis hypogaea) seedling roots, when expressed in Escherichia coli and insect cells (Sf9) gave a 29 kDa subunit protein. The native recombinant protein agglutinates neuraminidase treated human erythrocytes and the agglutination is inhibited by galactose. Nucleotide sequence and predicted amino acid sequence analyses indicate that it is different from peanut seed (PNA and SGL) and nodule (NGLa and NGLb) galactose-binding lectins. PMID- 11731093 TI - Evidence of very early neuronal migration from the olfactory placode of the chick embryo. AB - Cell migration from the olfactory neuroepithelium is a very peculiar phenomenon in the development of the nervous system. In this paper, we provide evidence that, in the chick embryo, migration of cells from the olfactory neuroepithelium begins earlier than previously reported, namely at the same time as the first olfactory placode differentiation, that occurs several hours before the superficial ectodermal invagination that gives rise to the olfactory pit. Moreover, we provide evidence that very early migrating cells express the HuC/D RNA-binding protein antigen, a specific neuronal marker. These observations refocus our knowledge on the very first developmental stages of olfactory neuroepithelium. PMID- 11731094 TI - Identification of hybrid cholinesterase forms consisting of acetyl- and butyrylcholinesterase subunits in human glioma. AB - Brain and non-brain tumors contain acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) transcripts and enzyme activity. AChE and BuChE occur in tissues as a set of molecular components, whose distribution in a cyst fluid from a human astrocytoma we investigated. The fluid displayed high BuChE and low AChE activities. Three types of cholinesterase (ChE) tetramers were identified in the fluid by means of sedimentation analyses and assays with specific inhibitors, and their sedimentation coefficients were 11.7S (ChE-I), 11.1S (ChE-II), and 10.5S (ChE-III). ChE-I was unretained, ChE-II was weakly retained and ChE-III was adsorbed to edrophonium-agarose, confirming the AChE nature of the latter. ChE-I and ChE-II tetramers contained BuChE subunits as shown by their binding with an antiserum against BuChE. The ChE activity of the immunocomplexes made with ChE-II and anti-BuChE antibodies decreased with the AChE inhibitor BW284c51, revealing that ChE-II was made of AChE and BuChE subunits, in contrast to ChE-I, which only contained BuChE subunits. The binding of an anti-AChE antibody (AE1) to ChE-II and ChE-III, but not to ChE-I, demonstrated the hybrid composition of ChE-II. A substantial fraction of the AChE tetramers and dimers of astrocytomas and oligodendrogliomas bound both to anti AChE and anti-BuChE antibodies, which revealed a mixed composition of AChE and BuChE subunits in them. The AChE components of brain, meningiomas and neurinomas were only recognized by AE1. In conclusion, our results demonstrate that aberrant ChE oligomers consisting of AChE and BuChE subunits are generated in astrocytomatous cyst and gliomas but not in brain, meningiomas or neurinomas. PMID- 11731095 TI - N-(2-Chloroethyl)-N-ethyl-2-bromobenzylamine reduces intracellular calcium response to noradrenaline in rat visual cortex. AB - Using the fluorescent indicator Fura-2, we investigated the effects of N-(2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4), a noradrenergic neurotoxin, on intracellular calcium responses to noradrenaline, N-methyl-D-aspartate, and carbamylcholine chloride in brain slices of the rat visual cortex. Noradrenergic depletion in the visual cortex of young rats was induced by DSP-4, and its selectivity was confirmed by two different methods, i.e., immunostaining with anti-dopamine-beta-hydroxylase antibody and biochemical analysis by high performance liquid chromatography. The treatment with DSP-4 (25 mg/kg i.p., x2) caused disruption of noradrenergic fibers throughout all cortical layers, and reduced the content of noradrenaline to 6.4% of that in the normal control. In the normal cortex, bath-applied noradrenaline (100 microM) increased the intracellular calcium to 123% of the control in terms of the F(340)/F(380) ratio of Fura-2 fluorescence. Quantitative analysis of the F(340)/F(380) ratio was performed in layers II to IV, since the increase was mainly observed in these layers. The intracellular calcium response to noradrenaline was significantly (P<0.0001) reduced in the DSP-4-treated animals to 63.2% of that in the normal control. The response to N-methyl-D-aspartate (100 microM) was also reduced, whereas the response to carbamylcholine chloride, a muscarinic cholinergic agonist (100 microM), was not affected by the DSP-4 treatment. From these findings we suggest that noradrenergic denervation by DSP-4 reduces the intracellular calcium response to noradrenaline through changes in the intracellular signal transduction. PMID- 11731096 TI - Physical activity elicits sustained activation of the cyclic AMP response element binding protein and mitogen-activated protein kinase in the rat hippocampus. AB - To elucidate molecular mechanisms involved in physical activity-induced beneficial effects on brain function, we studied in rats the influence of voluntary running on the activation in the hippocampus of cyclic AMP response element-binding protein (CREB) and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK). These are signaling molecules that play critical roles in synaptic plasticity, including learning and memory. Exercise resulted in an increase in the level of the activated transcription factor, CREB phosphorylated at Ser-133. The amount of the activated transcription factor about doubled already after 1 night of running and remained elevated for at least a week, although control levels were restored after 1 month of exercise. In addition, binding activity in nuclear extracts to cyclic AMP response element (CRE) motif containing oligonucleotides increased significantly in the hippocampus after 3 nights of exercise, although the total amount of the immunochemically identified CREB remained unaltered. Electrophoretic mobility supershift assays indicated that the increased binding was due to the recruitment of members of this transcription factor family, in addition to the CREB proper. Voluntary running also resulted in an increase in the level of phosphorylated MAPK (both p42 and p44). The time-courses of the increases in the level of the phosphorylated protein kinase and the activated transcription factor were different. In comparison with the activated CREB, the increase in the phosphorylated MAPK was delayed, but lasted longer, being detectable even after 1 month of exercise. These observations are consistent with the view that the relatively long-lasting activation of these signaling molecules participates in the regulation of genes, such as the neurotrophin genes, and contributes to the beneficial effects of physical exercise on brain function. PMID- 11731097 TI - Effects of age and insulin-like growth factor-1 on neuron and synapse numbers in area CA3 of hippocampus. AB - Age-related effects associated with the hippocampus include declines in numbers of neurons and synapses in the dentate gyrus and area CA1, and decreased cognitive ability as assessed with the Morris water maze. The present study quantified both neuron and synapse number in the same tissue block of area CA3 of the hippocampus. No investigations of both density of neurons and synapses together in area CA3 of hippocampus have been performed previously, despite its importance as the terminal field of dentate gyrus mossy fibers, the second synapse in the trisynaptic circuit in the hippocampus. Numerical density of neurons and synapses were assessed in 4-, 18-, and 29-month-old rats receiving infusions of saline into the lateral ventricle and in 29-month-old rats receiving infusions of insulin-like growth factor-1 (IGF-1). Numerical density of neurons of the stratum pyramidale of CA3 of hippocampus remained constant across the life span as did the numerical density of synapses in stratum lucidum of area CA3. Despite the reported role of IGF-1 in synaptogenesis and improvements in behavior with age, ventricular infusion of this growth factor did not affect the numerical density of neurons or synapses in 29-month-old rats when compared to saline infused old rats. Further, reported effects of IGF-1 on adult neurogenesis in the dentate gyrus are not reflected in an IGF-1-related increase in synapse density in this region. PMID- 11731098 TI - Non-spatial acquisition and retention deficits following small excitotoxic lesions within the hippocampus in monkeys. AB - Marmoset monkeys with excitotoxic lesions confined to cornu ammonis subfields 1 3, subiculum and pre-subiculum, but sparing the entorhinal cortex, were impaired on retention and learning of conditional object-choice discriminations. For each of these discriminations, the monkeys were required to choose one of two objects depending on which of two patterned backgrounds was used on each trial. Two styles of order of trial presentation were used: 'random' presentation which maximised the degree of interference between trials, and 'runs' presentation which was intended to encourage the monkeys to learn each component of the discrimination separately. Before surgery monkeys found the discriminations more difficult to learn when the trials were presented in the 'runs' style than when presented in the 'random' style suggesting that the task is best learnt by applying a conditional rule. After surgery a significant 'group x style' interaction indicated that the 'runs' style was especially difficult for the lesioned monkeys. From these results we suggest that the hippocampus is involved in learning about and remembering non-spatial, conditional relations between objects. PMID- 11731099 TI - Evidence for glutamate, in addition to acetylcholine and GABA, neurotransmitter synthesis in basal forebrain neurons projecting to the entorhinal cortex. AB - Basal forebrain neurons play important parts in processes of cortical activation and memory that have been attributed to the cortically projecting, cholinergic neurons. Yet, non-cholinergic neurons also project to the cerebral cortex and also appear to participate in processes of cortical modulation and plasticity. GABAergic neurons compose a portion of the cortically projecting cell group, but do not fully account for the non-cholinergic cell contingent. In the present study in the rat, we investigated whether the non-cholinergic, non-GABAergic cell component might be composed of glutamatergic neurons. We examined afferents to the entorhinal cortex, which is known to be modulated by basal forebrain neurons and to be critically involved in memory. Dual immunofluorescent staining was performed for cholera toxin, as retrograde tracer, and phosphate-activated glutaminase, the synthetic enzyme for the neurotransmitter pool of glutamate. The retrogradely labeled cells were distributed across the basal forebrain through the medial septum, diagonal band, magnocellular preoptic area and substantia innominata. The major proportion (approximately 80%) of the retrogradely labeled cells was found to be immunopositive for phosphate-activated glutaminase. Equal minor proportions (approximately 40%) were immunopositive for choline acetyltransferase and glutamic acid decarboxylase. In other material dual immunostained for neurotransmitter enzymes, approximately 95% of choline acetyltransferase- and approximately 60% of glutamic acid decarboxylase immunopositive neurons were also immunopositive for phosphate-activated glutaminase. From these results it appears that a significant proportion of these cell groups, including their cortically projecting contingents, could synthesize glutamate together with acetylcholine or GABA as neurotransmitters and another proportion of cells could synthesize glutamate alone. Accordingly, as either co transmitter or primary transmitter within basalocortical afferents, glutamate could have the capacity to modulate the entorhinal cortex and promote its role in memory. PMID- 11731100 TI - Amphetamine-induced toxicity in dopamine terminals in CD-1 and C57BL/6J mice: complex roles for oxygen-based species and temperature regulation. AB - In order to examine differential strain susceptibility to neurotoxic effects of amphetamine and to assess the potential role of superoxide radicals in amphetamine-induced dopaminergic damage, the drug was injected to mice with different levels of copper/zinc superoxide dismutase (Cu/Zn SOD) enzyme. Administration of amphetamine (10 mg/kg, i.p., given every 2 h, a total of four times) to wild-type CD-1 and C57BL/6J mice caused significant decreases in dopamine and 3,4-dihydroxyphenylacetic acid levels, in [(125)I]RTI-121-labeled dopamine transporters as well as a significant depletion in the concentration of dopamine transporter and vesicular monoamine transporter 2 proteins. The amphetamine-induced toxic effects were less prominent in CD-1 mice, which have much higher levels of Cu/Zn SOD activity (0.69 units/mg of protein) in their striata than C57BL/6J animals (0.007 units/mg of protein). Transgenic mice on CD 1 and C57BL/6J background, which had striatal levels of Cu/Zn SOD 2.57 and 1.67 units/mg of protein, respectively, showed significant protection against all the toxic effects of amphetamine. The attenuation of toxicity observed in transgenic mice was not caused by differences in amphetamine accumulation in wild-type and mutant animals. However, CD-1-SOD transgenic mice showed marked hypothermia to amphetamine whereas C57-SOD transgenic mice did not show a consistent thermic response to the drug. The data obtained demonstrate distinctions in the neurotoxic profile of amphetamine in CD-1 and C57BL/6J mice, which show some differences in Cu/Zn SOD activity and in their thermic responses to amphetamine administration. Thus, these observations provide evidence for possible complex interactions between thermoregulation and free radical load in the long-term neurotoxic effects of this illicit drug of abuse. PMID- 11731101 TI - Bidirectional actions of nociceptin/orphanin FQ on A delta-fibre-evoked responses in rat superficial spinal dorsal horn in vitro. AB - The present study investigated the modulatory actions of nociceptin/orphanin FQ on excitatory glutamatergic transmission in spinal dorsal horn. In transverse spinal cord slices with an attached dorsal root, mono- and polysynaptic A delta fibre-evoked extracellular field potentials were recorded from superficial dorsal horn. Nociceptin/orphanin FQ showed bidirectional effects on monosynaptic transmission with a potentiation at lower concentrations (100-300 nM) and a dose dependent depression at higher concentrations (1-3 microM). The polysynaptic field potential was dose-dependently depressed by nociceptin/orphanin FQ (100 nM 3 microM). None of the actions of nociceptin/orphanin FQ was reversed by the non specific opioid receptor antagonist naloxone, the N-methyl-D-aspartate receptor antagonist D-2-amino-5-phosphonovaleric acid or the peptide nocistatin. The bidirectional actions of nociceptin/orphanin FQ on the monosynaptic field potential may provide an in vitro model for the bidirectional actions of nociceptin/orphanin FQ in behavioural studies showing hyperalgesia at low doses of intrathecal nociceptin/orphanin FQ and analgesia at higher doses. PMID- 11731102 TI - Pre- and postsynaptic actions of ATP on neurotransmission in rat submandibular ganglia. AB - The pre- and postsynaptic actions of exogenously applied ATP were investigated in intact and dissociated parasympathetic neurones of rat submandibular ganglia. Nerve-evoked excitatory postsynaptic potentials (EPSPs) were not inhibited by the purinergic receptor antagonists, suramin and pyridoxal-phosphate-6-azophenyl 2',4'-disulphonic acid (PPADS), or the desensitising agonist, alpha,beta methylene ATP. In contrast, EPSPs were abolished by the nicotinic acetylcholine receptor antagonists, hexamethonium and mecamylamine. Focal application of ATP (100 microM) had no effect on membrane potential of the postsynaptic neurone or on the amplitude of spontaneous EPSPs. Taken together, these results suggest the absence of functional purinergic (P2) receptors on the postganglionic neurone in situ. In contrast, focally applied ATP (100 microM) reversibly inhibited nerve evoked EPSPs. Similarly, bath application of the non-hydrolysable analogue of ATP, ATP gamma S, reversibly depressed EPSPs amplitude. The inhibitory effects of ATP and ATP gamma S on nerve-evoked transmitter release were antagonised by bath application of either PPADS or suramin, suggesting ATP activates a presynaptic P2 purinoceptor to inhibit acetylcholine release from preganglionic nerves in the submandibular ganglia. In acutely dissociated postganglionic neurones from rat submandibular ganglia, focal application of ATP (100 microM) evoked an inward current and subsequent excitatory response and action potential firing, which was reversibly inhibited by PPADS (10 microM). The expression of P2X purinoceptors in wholemount and dissociated submandibular ganglion neurones was examined using polyclonal antibodies raised against the extracellular domain of six P2X purinoceptor subtypes (P2X(1-6)). In intact wholemount preparations, only the P2X(5) purinoceptor subtype was found to be expressed in the submandibular ganglion neurones and no P2X immunoreactivity was detected in the nerve fibres innervating the ganglion. Surprisingly, in dissociated submandibular ganglion neurones, high levels of P2X(2) and P2X(4) purinoceptors immunoreactivity were found on the cell surface. This increase in expression of P2X(2) and P2X(4) purinoceptors in dissociated submandibular neurones could explain the increased responsiveness of the neurones to exogenous ATP. We conclude that disruption of ganglionic transmission in vivo by either nerve damage or synaptic blockade may up-regulate P2X expression or availability and alter neuronal excitability. PMID- 11731103 TI - Depletion of substance P from rat primary sensory neurons by ATP, an implication of P2X receptor-mediated release of substance P. AB - Effects of ATP on substance P immunoreactivity were examined in cultured dorsal root ganglion neurons. We found that treatment of dorsal root ganglion neurons with ATP significantly depleted substance P immunoreactivity on the neurites and somata of the neurons. The effects of ATP were significantly inhibited by the purinergic P2 receptor antagonists suramin (30 microM) and pyridoxal-phosphate-6 azophenyl-2',4'-disulfonic acid (10 microM). We also showed that ATP-induced depletion of substance P immunoreactivity from dorsal root ganglion neurons depended on the entry of Ca(2+). In a spinal cord slice preparation, we also found the internalization of neurokinin-1/substance P receptors in many dorsal horn neurons following the application of ATP or alpha,beta-methylene-ATP. Together these results indicate that activation of P2X receptors may result in release of substance P from primary afferent neurons. PMID- 11731104 TI - Expression of brain-derived neurotrophic factor in rat dorsal root ganglia, spinal cord and gracile nuclei in experimental models of neuropathic pain. AB - Chronic constriction injury of the sciatic nerve and lumbar L5 and L6 spinal nerve ligation provide animal models for pain syndromes accompanying peripheral nerve injury and disease. In the present study, we evaluated changes in brain derived neurotrophic factor (BDNF) immunoreactivity in the rat L4 and L5 dorsal root ganglia (DRG) and areas where afferents from the DRG terminates (the L4/5 spinal cord and gracile nuclei) in these experimental models of neuropathic pain. Chronic constriction injury induced significant increase in the percentage of small, medium and large BDNF-immunoreactive neurons in the ipsilateral L4 and L5 DRG. Following spinal nerve ligation, the percentage of large BDNF-immunoreactive neurons increased significantly, and that of small BDNF-immunoreactive neurons decreased markedly in the ipsilateral L5 DRG, while that of BDNF-immunoreactive L4 DRG neurons of all sizes showed marked increase. Both chronic constriction injury and spinal nerve ligation induced significant increase in the number of BDNF-immunoreactive axonal fibers in the superficial and deeper laminae of the L4/5 dorsal horn and the gracile nuclei on the ipsilateral side. Considering that BDNF may modulate nociceptive sensory inputs and that injection of antiserum to BDNF significantly reduces the sympathetic sprouting in the DRG and allodynic response following sciatic nerve injury, our results also may suggest that endogenous BDNF plays an important role in the induction of neuropathic pain after chronic constriction injury and spinal nerve ligation. In addition, the increase of BDNF in L4 DRG may contribute to evoked pain which is known to be mediated by input from intact afferent from L4 DRG following L5 and L6 spinal nerve ligation. PMID- 11731105 TI - Superficial and deep convergent nociceptive neurons are differentially affected by N-methyl-D-aspartate applied on the brainstem surface of the rat medullary dorsal horn. AB - The activation of N-methyl-D-aspartate receptors is implicated in the spinal and trigeminal processing of nociceptive information conveyed by convergent (wide dynamic range) neurons and particularly in C-fiber-evoked responses elicited by repetitive and high-intensity electrical stimulation of the neuronal receptive field. In this study, the effects of intrathecal NMDA application on the electrically evoked nociceptive responses of trigeminal subnucleus caudalis convergent neurons have been investigated. The total C-fiber-evoked activity triggered by 30 successive stimuli was divided into two components: the C-fiber input response and the 'wind-up' response. Application of 0.1 microg (in 50 microl) of NMDA evoked a bi-directional effect on the total C-fiber-evoked activity of 19 neurons tested. A significant increase in the total C-fiber-evoked activity was observed 15-25 min after the NMDA application for nine neurons located in superficial laminae II and III. In contrast, a significant decrease in the total C-fiber-evoked activity was observed 5-25 min after the NMDA application for 10 neurons located more deeply, in lamina V. The NMDA-induced modifications seen in the total C-fiber-evoked activity were likely a reflection of a significant increase or decrease in neuronal activity evoked by the C-fiber input rather than wind-up of the responses since the latter was not significantly modified by the NMDA application. These results provide evidence for a possible inhibitory role for NMDA-dependent interneurons of the superficial laminae of the medullary dorsal horn on the nociceptive activity of deep convergent neurons. PMID- 11731106 TI - Low doses of N-methyl-D-aspartate antagonists in superficial laminae of medulla oblongata facilitate wind-up of convergent neurones. AB - In this study, a trigeminal model was used in which high threshold C-fibre-evoked activities of convergent neurones located in the subnucleus oralis of the trigeminal complex are modulated through the superficial part, the substantia gelatinosa, of the subnucleus caudalis. The two subnuclei are located 3 mm apart, therefore, it was possible to inject dizocilpine, a non-competitive N-methyl-D aspartate (NMDA) receptor antagonist, into either the superficial or the deep parts of subnucleus caudalis without interfering with ongoing recording of convergent neurones in subnucleus oralis. A differential NMDA-dependent modulation of wind-up was observed according to the dose and the injection target. (1) The injections of small non-diffusible doses (0.12 microg) of dizocilpine into the superficial part of subnucleus caudalis facilitated wind-up. The effect peaked at 25 min with a mean increase above control of 173+/-31%. Injection (0.5 microg) of either the less active enantiomer dizocilpine or saline into superficial subnucleus caudalis had no significant effect on subnucleus oralis convergent neurones. This suggests that NMDA-dependent interneurones, probably located in substantia gelatinosa of subnucleus caudalis, exert, in normal conditions, an inhibitory control on wind-up of convergent subnucleus oralis neurones. (2) The injection of larger doses (0.5 microg) into the superficial part of subnucleus caudalis induced a predominant inhibitory effect on wind-up. The mean peak effect at 15 min was 46+/-7% compared to control (100%). Small and large doses of dizocilpine injected into the deep part of subnucleus caudalis had a predominant inhibitory effect. The inhibition of wind up of subnucleus oralis neurones after injection of NMDA receptor antagonists in superficial or deep subnucleus caudalis indicates that wind-up may be due, at least in part, to NMDA activation at synapses that do not involve the recorded convergent neurones. PMID- 11731107 TI - Selective depression of nociceptive responses of dorsal horn neurones by SNC 80 in a perfused hindquarter preparation of adult mouse. AB - Detailed electrophysiological characterisation of spinal opioid receptors in the mouse has been limited due to various technical difficulties. In this study, extracellular single unit recordings were made from dorsal horn neurones in a perfused spinal cord with attached trunk-hindquarter to investigate the role of delta-opioid receptor in mediating nociceptive and non-nociceptive transmission in mouse. Noxious electrical shock, pinch and heat stimuli evoked a mean response of 20.8+/-2.5 (n=10, P<0.005), 30.1+/-5.4 (n=58, P<0.005) and 40.9+/-6.3 (n=29, P<0.005) spikes per stimulus respectively. In 5 of 22 cells, repetitive noxious electrical stimuli applied to the hindpaw for 20 s produced a progressive increase in spike number, the phenomenon known as 'wind-up' and/or hyperactivity. When the selective delta-opioid receptor agonist (+)-4-[(alpha R)-alpha-((2S,5R) 4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide (SNC 80) was perfused for 8-10 min, these evoked nociceptive responses were reversibly depressed. SNC 80 (2 microM) depressed the nociceptive responses evoked by electrical shock, pinch and heat by 74.0+/-13.7% (n=8, P<0.01), 66.5+/-16.6% (n=10, P<0.01) and 74.1+/-17.0% (n=10, P<0.01) respectively. The maximum depression by 5 microM SNC 80 was 92.6+/-6.8% (n=3). SNC 80 at 5 microM also completely abolished the wind-up and/or hypersensitivity (n=5). The depressant effects of SNC 80 on the nociceptive responses were completely blocked by 10 microM naloxone (n=5) and 3 microM 17-(cyclopropylmethyl)-6,7-dehydro-4,5 alpha epoxy-14 beta-ethoxy-5 beta-methylindolo [2',3':6',7'] morphinan-3-ol hydrochloride (HS 378, n=8), a novel highly selective delta-opioid receptor antagonist. Interestingly, HS 378 (3 microM) itself potentiated the background activity and evoked responses to pinch and heat by 151.8+/-38.4% (P<0.05, n=8), 34.2+/-6.1% (P<0.01, n=7) and 45.5+/-11.8% (P<0.05, n=5) respectively. In contrast, the responses of non-nociceptive dorsal horn neurones were not inhibited by SNC 80 at a dose of up to 10 microM (n=5). These data demonstrate that delta-opioid receptor modulate nociceptive, but not non-nociceptive, transmission in spinal dorsal horn neurones of the adult mouse. The potentiation of neuronal activity by HS 378 may reflect an autoregulatory role of the endogenous delta-opioid in nociceptive transmission in mouse. PMID- 11731108 TI - Inhibitors of trypsin-like serine proteases prevent DNA damage-induced neuronal death by acting upstream of the mitochondrial checkpoint and of p53 induction. AB - We have previously shown that the pharmacological agents 4-(2 aminoethyl)=benzenesulfonylfluoride hydrochloride (AEBSF) and Na-p-tosyl-L-lysine chloromethylketone (TLCK), inhibitors of trypsin-like serine proteases, prevent the death of trophic factor-deprived PC12 cells and sympathetic neurons. Both AEBSF and TLCK inhibit caspase activation in this model, but it is unclear whether they do so indirectly or through a direct effect at the level of the caspases. In the current study, we have used these agents in another model of neuronal death that is induced by DNA damage. We find that both agents delay the death of DNA-damaged PC12 cells, neonatal rat sympathetic neurons and embryonic rat cortical neurons. As in the trophic deprivation model, they act upstream of the caspases. In addition, they prevent mitochondrial alterations, such as cytochrome c release or loss of transmembrane potential. In contrast, the general caspase inhibitor bok-asp-fmk does not prevent cytochrome c release and has only a partial and transient effect on loss of transmembrane potential. Interestingly, both AEBSF and TLCK prevent the induction and nuclear accumulation of p53 that is induced by DNA damage in cortical neurons. Therefore, these serine protease inhibitors act at a point upstream in the apoptotic pathway, prior to p53 induction and the mitochondrial checkpoint, to delay neuronal death in this model, and do not act at the level of the caspases. We conclude that therapeutic strategies based on serine protease inhibition may be useful in preventing neuronal cell death. PMID- 11731109 TI - ErbB-4 activation inhibits apoptosis in PC12 cells. AB - Neuregulins, a large family of polypeptide growth factors, exert various distinctive effects in the nervous system. neuregulins and their receptors are widely expressed in neurons implying important roles in neuronal cell functions. Recently, we have shown that ErbB-4 receptors expressed in PC12 cells mediate neuregulin-induced differentiation. In the present study we demonstrate that in the PC12-ErbB-4 cells, neuregulin rescues cells from apoptosis induced by serum deprivation or tumor necrosis factor (TNF)alpha treatment. The neuregulin-induced survival is comparable to the effect mediated by the neurotrophic factor nerve growth factor (NGF). Both neuregulin and NGF protect cells from apoptosis induced by serum deprivation and TNF alpha treatment. Moreover, neuregulin like NGF induces the survival of neuronal differentiated PC12-ErbB-4 cells. The survival effect of neuregulin is probably mediated by the phosphoinositide 3-kinase (PI3K) and protein kinase B/Akt signaling pathways. Neuregulin induces the activation of PI3K and prolonged activation of protein kinase B/Akt. In addition, inhibition of the PI3K activity prevented the neuregulin-induced survival effect. Taken together, these results indicate that survival induced by neuregulin in PC12-ErbB 4 cells requires PI3K signaling networks. PMID- 11731110 TI - Armillaramide, a new sphingolipid from the fungus Armillaria mellea. AB - A new C(18)-phytosphingosine ceramide containing non-hydroxy fatty acid, armillaramide (1), has been isolated together with ergosterol peroxide from the fruiting bodies of the basidiomycete Armillaria mellea. Its structure was established as (2S,3S,4R)-2-hexadecanoylamino-octadecane-1,3,4-triol by spectroscopic and chemical methods. PMID- 11731111 TI - Chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. AB - The chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch (HBP) carcinogenesis was examined using lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) as biomarkers of chemoprevention. Twenty four male Syrian hamsters were divided into four groups of six animals each. The right buccal pouches of the animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin three times a week. The animals in group 2 were painted with DMBA as in group 1 and in addition received 2.5 mg/kg body weight lycopene orally three times a week on days alternate to DMBA application. Group 3 animals received lycopene as in group 2. Animals in group 4 received neither DMBA nor lycopene and served as control. The hamsters were killed after an experimental period of 14 weeks. Biochemical measurements were carried out in tumour and normal tissues. All hamsters painted with DMBA alone for 14 weeks developed well-differentiated squamous cell carcinomas. Diminished lipid peroxidation in the oral tumour tissue was accompanied by a significant increase in the levels of GSH, GPx, GST and GR. Administration of lycopene significantly suppressed DMBA-induced oral carcinogenesis as revealed by the absence of carcinomas. The results of the present study suggest that lycopene may exert its chemopreventive effects by modulating lipid peroxidation and enhancing the activities of the enzymes in the glutathione redox cycle. PMID- 11731112 TI - Two novel proaporphine-tryptamine dimers from Roemeria hybrida. AB - Roemeria hybrida yielded two novel proaporphine-tryptamine N-oxides, (-) roehybridine alpha-N-oxide (1) and (-)-roehybramine beta-N-oxide (2). NMR data allowed a facile assignment of these proaporphine-tryptamine dimers into different stereochemical subgroups. PMID- 11731113 TI - An anti-inflammatory and immunomodulatory polysaccharide from Orbignya phalerata. AB - A polysaccharide, a glucan with mean M(r) of 1.0 x 10(6) (MP1), was isolated from the mesocarp of fruits of Orbignya phalerata. Chemical and spectroscopic studies indicated that MP1 has a highly branched glucan type structure composed of alpha (1-->4) linked D-glucopyranose residues with (3-->4), (4-->6), and with (3-->6) branching points. MP1 enhanced phagocytosis in vivo and exhibited anti inflammatory activity. PMID- 11731114 TI - Cycloartane saponins from Astragalus peregrinus as modulators of lymphocyte proliferation. AB - From Astragalus peregrinus, four cycloartane-type saponins have been isolated and their structures elucidated by spectral means as 20(R),24(S)-epoxy-9 beta,19 cyclolanostane-3 beta,6 alpha,16 beta,25-tetrol 3-O-beta-D-glucopyranoside (1), 20(R),24(S)-epoxy-9 beta,19-cyclolanostane-3 beta,6 alpha,16 beta,25-tetrol 3-O alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranoside (2), 20(R),24(S)-epoxy-9 beta,19-cyclolanostane-3 beta,6 alpha,16 beta,25-tetrol 3-O-alpha-L rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside (3) and 20(R),25-epoxy-9 beta,19 cyclolanostane-3 beta,6 alpha,16 beta,24(S)-tetrol (24-O-acetyl)- 3-O-alpha-L rhamnopyranosyl-(1-->2)-(6'-O-acetyl)-beta-D-glucopyranoside (4). Compounds 2 and 3 showed to stimulate the proliferation of mouse splenocytes and were not significantly cytotoxic. PMID- 11731115 TI - Effect of Spirulina fusiformis on cyclophosphamide and mitomycin-C induced genotoxicity and oxidative stress in mice. AB - Spirulina fusiformis was tested for its possible in vivo protective effects against cyclophosphamide (CP) and mitomycin-C (MMC) induced genotoxicity and oxidative stress in mice. Pre-treatment with S. fusiformis (250, 500 and 1000 mg kg(-1), p.o., daily for 5 days) significantly reduced the chromosomal damage and lipid peroxidation with concomitant changes in antioxidants and detoxification systems. All the three tested doses were effective in exerting a protective effect against CP and MMC. PMID- 11731116 TI - Artoindonesianin L, a new prenylated flavone with cytotoxic activity from Artocarpus rotunda. AB - A new prenylated flavone, named artoindonesianin L (1), was isolated from Artocarpus rotunda (Hout) Panzer (Moraceae). Its structure was elucidated as on the basis of spectroscopic evidence. Along with this new compound, four known phenolic compounds were also isolated from this plant and identified as artonins M (2) and E (3), cycloartobiloxanthone (4) and artonin O (5). All these compounds showed significant cytotoxicity against murine P388 leukemia cells. PMID- 11731117 TI - Analgesic sesquiterpene dilactone from Mikania cordata. AB - The crude extract of Mikania cordata (1 and 3 g/kg, p.o.) and deoxymikanolide (1) (10 mg/kg, p.o.) significantly inhibited acetic acid-induced writhing in mice. Three other sesquiterpene dilactones isolated from the same plant, namely mikanolide, dihydromikanolide and scandenolide, did not show significant analgesic activity. PMID- 11731118 TI - Anxiolytic activity of aerial and underground parts of Passiflora incarnata. AB - The petroleum ether, chloroform, methanol, and water extracts of Passiflora incarnata whole plant and sorted out plant parts have been evaluated for their anxiolytic activity using the elevated plus-maze model in mice. The methanol extracts of leaves, stems, flowers, and whole plant exhibited anxiolytic effects at 100, 125, 200 and 300 mg/kg, respectively. The roots were practically devoid of anxiolytic effects. These results show that roots and flowers of P. incarnata act as natural adulterants by causing a significant increase in the anxiolytic dose. Therefore, separation of these parts is recommended prior to any pharmacological, phytochemical and standardization studies on P. incarnata. PMID- 11731119 TI - Antibacterial activity of Urena lobata root. AB - The methanol extract of Urena lobata root showed a broad spectrum of antibacterial activity. PMID- 11731120 TI - Antimicrobial activity of Securinega leucopyrus. AB - Extracts of the leaves of Securinega leucopyrus exhibited a broad spectrum of antimicrobial activity. PMID- 11731121 TI - Phytochemical and antimicrobial studies on Drynaria quercifolia. AB - Friedelin, epifriedelinol, beta-amyrin, beta-sitosterol, beta-sitosterol 3-beta-D glucopyranoside, and naringin were isolated from the dried rhizome of Drynaria quercifolia. The methanol extract showed broad and concentration-dependent antibacterial activity. PMID- 11731122 TI - Hypolipidemic effects of Teucrium polium in rats. AB - The aqueous extract of Teucrium polium aerial parts, given intraperitoneally at doses from 50 to 150 mg/kg for 10 days, reduced significantly the serum levels of cholesterol and triglycerides in hyperlipidemic rats. PMID- 11731123 TI - Two flavonol 5-O-glycosides from the roots of Leuzea carthamoides. AB - Two new flavonoids, identified as quercetin 5-O-galactoside (1) and isorhamnetin 5-O-rhamnoside (2) were isolated from the roots of Leuzea carthamoides. PMID- 11731124 TI - A new C-methylated flavonoid glycoside from Pinus densiflora. AB - A new C-methyl flavonol glycoside, 5,7,8,4'-tetrahydroxy-3-methoxy-6 methylflavone 8-O-beta-D-glucopyranoside (1), has been isolated from the needles of Pinus densiflora, together with kaempferol 3-O-beta-(6"-acetyl) galactopyranoside. PMID- 11731125 TI - A new ent-kaurane diterpenoid from Rabdosia pseudo-irrorata. AB - A new kaurenoid, ent-3 beta,7 alpha,14 beta-trihydroxykaur-16-ene-12,15-dione (1), named pseurata G, was isolated from the ethanol extract of the whole plant of Rabdosia pseudo-irrorata. PMID- 11731126 TI - Super-channel in bacteria: function and structure of the macromolecule import system mediated by a pit-dependent ABC transporter. AB - In a soil isolate, Sphingomonas sp. A1, the transport of a macromolecule (alginate: 27 kDa) is mediated by a pit-dependent ATP-binding cassette (ABC) transporter. The transporter is different from other ABC transporters so far analyzed in that its function is dependent on a pit, a mouth-like organ formed on the cell surface only when cells are compelled to assimilate macromolecules, and in that it allows direct import of macromolecules into cells. The ABC transporter coupled with the pit, which functions as a funnel and/or concentrator of macromolecules to be imported, was designated the 'super-channel', and in this review, we discuss the three-dimensional structure and specific function of the 'super-channel' for macromolecule import found for the first time in a bacterium. PMID- 11731127 TI - Phylogenetic and structural analyses of the oxa1 family of protein translocases. AB - Mitochondrial Oxa1p homologs have been shown to function in protein export and membrane insertion in bacteria, mitochondria and chloroplasts, but their mode of action, organismal distribution and evolutionary origins are poorly understood. All sequenced homologs of Oxa1p were retrieved from the databases and multiply aligned. All organisms with a fully sequenced genome possess at least one Oxa1p homolog showing that the family is truly ubiquitous. Most prokaryotes possess just one Oxa1p homolog, but several Gram-positive bacteria and one archaeon possess two, and eukaryotes may have as many as six. Although these proteins vary in length over a 5-fold range, they exhibit a common hydrophobic core region of about 200 residues. Multiple sequence alignments reveal conserved residues and provide the basis for structural and phylogenetic analyses that serve to characterize the Oxa1 family. PMID- 11731128 TI - Two sequence classes of kinetoplastid 5S ribosomal RNA gene revealed among bodonid spliced leader RNA gene arrays. AB - The spliced leader RNA genes of Bodo saltans, Cryptobia helicis and Dimastigella trypaniformis were analyzed as molecular markers for additional taxa within the suborder Bodonina. The non-transcribed spacer regions were distinctive for each organism, and 5S rRNA genes were present in Bodo and Dimastigella but not in C. helicis. Two sequence classes of 5S rRNA were evident from analysis of the bodonid genes. The two classes of 5S rRNA genes were found in other Kinetoplastids independent of co-localization with the spliced leader RNA gene. PMID- 11731129 TI - A putative ATP-binding cassette transporter YbdA involved in sporulation of Bacillus subtilis. AB - Insertional mutagenesis with mini-Tn10 was performed to identify new genes involved in sporulation of Bacillus subtilis. Here, we report on the characterization of the ybdA locus, which encodes a putative ATP-binding cassette transporter. The ybdA gene is the 6th cistron of the putative ybcOPQST-ybdABDE operon. A deletion mutation in ybdA and an insertional mutation in ybdB exhibited highly oligosporogenous phenotypes and led to a decrease in the transcription controlled by Spo0A, which is a key response regulator required for the initiation of sporulation. We further observed that the transcription of this operon was strongly induced after the end of the exponential growth phase in the wild-type strain, but not in a spo0A null mutant. Our data suggest that the YbdA and YbdB proteins are able to affect incorporation of nutrient signals during initiation of sporulation and may act as components of positive feedback systems of Spo0A activation. PMID- 11731130 TI - Metabolic flux response to phosphoglucose isomerase knock-out in Escherichia coli and impact of overexpression of the soluble transhydrogenase UdhA. AB - Blocking glycolytic breakdown of glucose by inactivation of phosphoglucose isomerase (Pgi) in Escherichia coli led to a greatly reduced maximum specific growth rate. Examination of the operational catabolic pathways and their flux ratios using [U-(13)C(6)]glucose-labeling experiments and metabolic flux ratio analysis provide evidence for the pentose phosphate (PP) pathway as the primary route of glucose catabolism in the knock-out mutant. The resulting extensive flux through the PP pathway disturbs apparently the reducing power balance, since overexpression of the recently identified soluble transhydrogenase UdhA improves significantly the growth rate of the Pgi mutant. The presented results provide first evidence that UdhA restores the cellular redox balance by catalyzing electron transfer from NADPH to NADH. PMID- 11731131 TI - Two new thermostable type II restriction endonucleases from Thermus aquaticus: TatI and TauI, which recognize the novel nucleotide sequences 5'-W (downward arrow)GTACW-3' and 5'-GCSG (downward arrow)C-3' respectively. AB - One hundred and forty isolates of thermophilic bacteria from the genus Thermus were screened for the presence of restriction endonuclease activity. Thermostable isoschizomers of restriction endonucleases, such as AceIII, BbvI, BglI, BsePI, FnuDII, HgiAI, MaeII, MboI, MseI, PvuII, StuI, TaqI, Tsp4CI, TspEI, XhoI and XmaIII, were isolated. Two restriction enzymes, TatI and TauI, recognizing novel degenerate sequences 5'-W (downward arrow)GTACW-3' and 5'-GCSG (downward arrow)C 3' respectively were partially purified and the recognition and cleavage sites were determined. PMID- 11731133 TI - The antibacterial action of microcin J25: evidence for disruption of cytoplasmic membrane energization in Salmonella newport. AB - Microcin J25 (MccJ25) is a cyclic peptide of 21 unmodified amino acid residues produced by a fecal strain of Escherichia coli. It has previously been shown that the antibiotic activity of this peptide is mainly directed to Enterobacteriaceae, including several pathogenic E. coli, Salmonella and Shigella strains. In this paper we show that MccJ25 acts on the cytoplasmic membrane of Salmonella newport cells producing alteration of membrane permeability, and the subsequent gradient dissipation, that initiate the inhibition of process, such as oxygen consumption. These results, taken together with our in vitro observations [Rintoul et al. (2000) Biochim. Biophys. Acta 1509, 65-72], strongly suggest that the disruption of the cytoplasmic membrane gradient is closely related to the bactericidal activity of MccJ25 in S. newport. PMID- 11731132 TI - Proteins of the lactococcin A secretion system: lcnD encodes two in-frame proteins. AB - Polyclonal antibodies were raised against LcnC and LcnD proteins of the Lactococcus lactis bacteriocin lactococcin A secretory system to examine their cellular location and interaction. Two major reacting bands were detected by Western immunoblot with the anti-LcnD antibody: one of 52 kDa (LcnD) and another of 45 kDa, called here LcnD*. LcnD* was still detectable after removing the AUG start codon for LcnD. Chemical cross-linking analyses of membrane fractions of L. lactis cells expressing the LcnC/D secretion machinery were performed. Our results indicate that LcnD is present in the secretion machinery complex as a dimer and is able to interact with LcnD* and LcnC. PMID- 11731134 TI - NADH dehydrogenase of Corynebacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH. AB - NADPH oxidase activity, in addition to NADH oxidase activity, has been shown to be present in the respiratory chain of Corynebacterium glutamicum. In this study, we tried to purify NADPH oxidase and NADH dehydrogenase activities from the membranes of C. glutamicum. Both the enzyme activities were simultaneously purified in the same fraction, and the purified enzyme was shown to be a single polypeptide of 55 kDa. The N-terminal sequence of the enzyme was consistent with the sequence deduced from the NADH dehydrogenase gene of C. glutamicum, which has been sequenced and shown to be a homolog of NADH dehydrogenase II. In addition to high NADH-ubiquinone-1 oxidoreductase activity at neutral pH, the purified enzyme showed relatively high NADPH oxidase and NADPH-ubiquinone-1 oxidoreductase activities at acidic pH. Thus, NADH dehydrogenase of C. glutamicum was shown to be rather unique in having a relatively high reactivity toward NADPH. PMID- 11731135 TI - A fluorescence quenching test for the detection of flavonoid transformation. AB - A novel fluorescence quenching test for the detection of flavonoid degradation by microorganisms was developed. The test is based on the ability of the flavonoids to quench the fluorescence of 1,6-diphenyl-1,3,5-hexatriene (DPH). Several members of the anthocyanidins, flavones, isoflavones, flavonols, flavanones, dihydroflavanones, chalcones, dihydrochalcones and catechins were tested with regard to their quenching properties. The anthocyanidins were the most potent quenchers of DPH fluorescence, while the flavanones, dihydroflavanones and dihydrochalcones, quenched the fluorescence only weakly. The catechins had no visible impact on DPH fluorescence. The developed test allows a quick and easy differentiation between flavonoid-degrading and flavonoid-non-degrading bacteria. The investigation of individual reactions of flavonoid transformation with the developed test system is also possible. PMID- 11731136 TI - Secretion of the Escherichia coli K-12 SheA hemolysin is independent of its cytolytic activity. AB - The Escherichia coli K-12 sheA gene encodes a pore-forming hemolysin that is secreted to the medium by a hitherto unidentified mechanism. To study SheA secretion, we constructed fusions between SheA and the mature form of the periplasmic enzyme beta-lactamase, and performed site-directed mutagenesis on these constructs. The SheA-Bla and Bla-SheA hybrid proteins displayed hemolytic activity and were efficiently exported to the extracellular medium. Our results with mutant hybrid proteins show that secretion of SheA is independent of its cytolytic activity, that secretion is paralleled by a transient leakage of periplasmic contents to the extracellular medium, and that deletion of the 11 C terminal residues of SheA has no effect on its secretion and cytolytic activity. PMID- 11731137 TI - Contribution of flagella and invasion proteins to pathogenesis of Salmonella enterica serovar enteritidis in chicks. AB - To explore the relative contribution that flagella and Salmonella invasion proteins make to the virulence of Salmonella enteritidis in poultry, 20-day-old chicks were challenged orally and by subcutaneous injection with wild-type strain SE-HCD, two non-flagellated mutants (fliC::Tn10 mutant and flhD::Tn10 mutant) and two Salmonella invasion protein insertion mutants (sipD and iacP). When injected subcutaneously, wild-type SE-HCD was the only strain to cause substantial mortality and morbidity and to grow well in organs. The flhD mutant of SE-HCD was invasive when given orally, whereas wild-type SE-HCD and the fliC mutant were significantly attenuated. Salmonella invasion protein mutants were not invasive by either route. These results suggest that temporary suppression of Class I regulators of flagellin biosynthesis may aid oral infection in poultry. PMID- 11731138 TI - Actinobacillus actinomycetemcomitans possesses an antigen binding to anti-human IL-10 antibody. AB - It is well known that the cell components of periodontopathic bacteria are able to induce several cytokines and possibly to affect the cytokine network. In order to determine the presence of the periodontopathic Actinobacillus actinomycetemcomitans components recognized by antibodies against cytokine molecules, ELISA reactivities of sonic extracts from the bacterial cells were determined by use of ELISA kits specific for human interleukin (IL)-1beta, IL-4, IL-5, IL-6, IL-10, tumor necrosis factor-alpha, and interferon-gamma. The ELISA analysis demonstrated that the sonic extracts from eight strains of A. actinomycetemcomitans bound with anti-human IL-10 monoclonal antibody. Western blotting analysis revealed that the molecular mass of the antigen was approximately 65 kDa. IL-10 is produced by type 2 helper T cells and mainly down regulates the type 1 helper T cell response. The present study suggests that the 65-kDa antigen of A. actinomycetemcomitans may affect the host defense function through binding to IL-10 receptor as an agonist or an antagonist for IL-10. PMID- 11731139 TI - Biodegradation of p-toluenesulphonamide by a Pseudomonas sp. AB - A bacterium capable of utilising p-toluenesulphonamide was isolated from activated sludge. The isolated strain designated PTSA was identified as a Pseudomonas sp. using chemotaxonomic and genetic studies. Pseudomonas PTSA grew on p-toluenesulphonamide in a chemostat with approximately 90% release of sulphate and 80% release of ammonium. The isolate was also able to grow on 4 carboxybenzenesulphonamide and 3,4-dihydroxybenzoate but did not grow on p toluenesulphonate. The transient appearance of 4-hydroxymethylbenzenesulphonamide and 4-carboxybenzenesulphonamide during p-toluenesulphonamide degradation proves oxidation of the methyl group is the initial attack in the biodegradation pathway. Both metabolites of p-toluenesulphonamide degradation were identified by high-performance liquid chromatography-mass spectrometry. 4 Carboxybenzenesulphonamide is probably converted into 3,4-dihydroxybenzoate and amidosulphurous acid. The latter is a chemically unstable compound in aqueous solutions and immediately converted into sulphite and ammonium. Both sulphite and ammonium were formed during degradation of 4-carboxybenzenesulphonamide. PMID- 11731140 TI - Effect of pheromone induction on transfer of the Enterococcus faecalis plasmid pCF10 in intestinal mucus ex vivo. AB - The effect of synthetic sex pheromone on pheromone-inducible conjugation between the isogenic Enterococcus faecalis strains OG1RF and OG1SS was investigated in (i) Todd-Hewitt broth medium and (ii) intestinal mucus isolated from germ-free rats. In broth, the presence of synthetic pheromone cCF10 had no detectable effect on the transfer kinetics observed for the tetracycline resistance encoding plasmid pCF10. In mucus, presence of the same pheromone significantly increased the transfer efficiency observed during the first 2 h of conjugation, while the effect was less pronounced later in the experiment. We suggest that due to differences in diffusion rates and medium-binding of the pheromones, the effect of the synthetic cCF10 was immediately dominated by the effect of pheromones produced by the recipient E. faecalis strain in broth, while this happened later in mucus. PMID- 11731141 TI - Nutritional influence on fungal colony growth and biomass distribution in response to toxic metals. AB - This work examines nutritional influence on fungal colony growth and biomass distribution in response to toxic metals. In low-substrate solid medium, 0.1 mM Cd, Cu and Zn caused a decrease in radial expansion of both Trichoderma viride and Rhizopus arrhizus. However, as the amount of available carbon source (glucose) increased, the apparent toxicity of the metals decreased. These metals also affected the overall length of the fungal mycelium and branching patterns. In low-nutrient conditions, T. viride showed a decrease in overall mycelial length and number of branches in response to Cu, resulting in an extremely sparsely branched colony. Conversely, although Cd also reduced overall mycelial length to about one-third of the control length, the number of branches decreased only slightly which resulted in a highly branched colony with many aberrant features. Cu and Cd induced similar morphological changes in R. arrhizus. A large scale mycelial-mapping technique showed that disruption of normal growth by Cu and Cd resulted in altered biomass distribution within the colony. When grown on metal-free low-substrate medium, T. viride showed an even distribution of biomass within the colony with some allocation to the periphery. However, Cu caused most of the biomass to be allocated to the colony periphery, while in the presence of Cd, most biomass was located at the interior of the colony. These results imply that such alterations of growth and resource allocation by Cu and Cd may influence success in locating nutrients as well as survival, and that these metals have individual and specific effects on the growing fungus. PMID- 11731142 TI - Following the course of human leptospirosis: evidence of a critical threshold for the vital prognosis using a quantitative PCR assay. AB - In order to follow the course of acute human leptospirosis, an ELISA microtiter plate hybridization method was developed for the quantitative determination of Leptospira spp. in biological samples after PCR. The biotin-labelled amplified product (331 bp from the rrs gene) was hybridized with a complementary capture probe covalently linked onto aminated polystyrene wells, and detected using a colorimetric reaction. The mean detection limit was 50 copies per 10 microl. In a prospective study of human leptospirosis cases, we obtained evidence that a density of 10(4) leptospires per ml of blood is a critical threshold for the vital prognosis of the patients. The practicability of the method makes it suitable for use in tropical areas for multicentric studies. Such studies could lead to a better knowledge of the natural history of the human disease. The method is also suitable for experimental evaluation of improved antibiotic treatments for leptospirosis. PMID- 11731143 TI - Ura-status-dependent adhesion of Candida albicans mutants. AB - Gene disruptions in the diploid opportunistic human fungal pathogen Candida albicans are usually created using multiple rounds of targeted integration called the 'ura-blaster' method. Resulting heterozygous and homozygous null mutants can be auxotrophic (Ura(-)) or prototrophic (Ura(+)) for uracil biosynthesis. Here we demonstrate that the Ura-status of otherwise isogenic mutants affected the adhesion of C. albicans. Moreover the effect of Ura-status on adhesion was also dependent on the null mutant background, the nature of the underlying surface and the carbon source for growth. Therefore the Ura-status is not neutral in determining adhesive properties of C. albicans mutants that are generated via the ura-blaster protocol. PMID- 11731144 TI - Sensitivity of Shiga toxin-producing Escherichia coli (STEC) strains for colicins under different experimental conditions. AB - Twenty Escherichia coli strains producing well-characterised colicins were tested for their inhibitory activity against five Shiga toxin-producing E. coli (STEC) strains using different media under aerobic and anaerobic conditions. The five STEC strains used were of serotype O26, O111, O128, O145 and O157:H7 which are frequently isolated serotypes associated with disease in humans. The main route of infection for humans is through the eating of badly cooked or handled beef. The major reservoir for STEC strains in cattle is the rumen. To mimic the situation in the rumen of cattle, overlay assays were also performed under anaerobic conditions in the presence of 30% rumen fluid. Colicins E1, E4, E8-J, K and S4 are most active against STEC strains under anaerobic conditions in the absence or presence of rumen fluid. These colicins will be used in future experiments with the aim to eradicate the presence of STEC in cattle. PMID- 11731145 TI - Arsenite oxidation and arsenate respiration by a new Thermus isolate. AB - A new microbial strain was isolated from an arsenic-rich terrestrial geothermal environment. The isolate, designated HR13, was identified as a Thermus species based on 16S rDNA phylogenetic relationships and close sequence similarity within the Thermus genus. Under aerobic conditions, Thermus HR13 was capable of rapidly oxidizing inorganic As(III) to As(V). As(III) was oxidized at a rate approximately 100-fold greater than abiotic rates. Metabolic energy was not gained from the oxidation reaction. In the absence of oxygen, Thermus HR13 grew by As(V) respiration coupled with lactate oxidation. The ability to oxidize and reduce arsenic has not been previously described within the Thermus genus. PMID- 11731146 TI - Lithoautotrophic growth of the freshwater strain Beggiatoa D-402 and energy conservation in a homogeneous culture under microoxic conditions. AB - The freshwater filamentous bacterium Beggiatoa D-402 was shown to grow lithoautotrophically in a homogeneous culture under microoxic conditions only, the growth yield being the highest at 0.1 mg O(2) l(-1). High activities of the Calvin cycle key enzymes and of the dissimilatory path thiosulfate oxidation enzymes were found in the bacterial cells. The rate of CO(2) fixation above 112 nmol min(-1) (mg protein)(-1), an about 90% increase in the protein carbon at the expense of CO(2) carbon and an increase in the molar yield up to 12 mg dry weight (mmol oxidized thiosulfate)(-1) indicate the bacterial growth was autotrophic. Thiosulfate was oxidized by the strain almost completely into sulfate. The metabolically useful energy was conserved by oxidative phosphorylation that was coupled to oxidation of sulfur compounds. The bacterial membranes were found to contain CO-binding cytochromes b and two cytochromes c with M(r) 23 and 26 kDa, the terminal part of the respiratory chain containing presumably a cbb(3)-type oxidase. A cytochrome c with M(r) 12 kDa was detected in the soluble fraction. PMID- 11731147 TI - Zeaxanthin and menaquinone-7 biosynthesis in Sphingobacterium multivorum via the methylerythritol phosphate pathway. AB - Feeding of [1-(13)C]glucose, [U-(13)C(6)]glucose, [3-(13)C]alanine and [1 (13)C]acetate to Sphingobacterium multivorum showed that this bacterium utilizes the methylerythritol phosphate pathway for the biosynthesis of menaquinone-7 and zeaxanthin, a carotenoid of industrial importance. Differential incorporation of the labeled precursors gave some insight into the preferred carbon sources involved in isoprenoid biosynthesis. PMID- 11731148 TI - The effect of co-overproduction of DnaK/DnaJ/GrpE and ClpB proteins on the removal of heat-aggregated proteins from Escherichia coli DeltaclpB mutant cells- new insight into the role of Hsp70 in a functional cooperation with Hsp100. AB - The effect of overproduction of the Hsp70 system proteins (DnaK, DnaJ, GrpE) and/or ClpB (Hsp100) from plasmids on the process of formation and removal of heat-aggregated proteins from Escherichia coli cells (the S fraction) was investigated by sucrose density gradient centrifugation. Two plasmids were employed: pKJE7 carrying the dnaK/dnaJ/grpE genes under the control of the araB promoter and pClpB carrying the clpB gene under the control of its own promoter (sigma(32)-dependent). In the wild-type cells the S fraction after 15 min of heat shock amounted to 21% of cellular insoluble proteins (IP), and disappeared 10 min after transfer of the culture to 37 degrees C. In contrast to this, in the clpB mutant the S fraction was larger (35% IP) and its elimination was retarded, nearly 60% of the aggregated proteins remained stable 30 min after heat shock. This result points to the importance of ClpB in removal of the heat-aggregated proteins from cells. Overproduction of the Hsp70 system proteins (exceeding by about 1.5-fold that of wild-type) in wild-type and DeltaclpB cells completely prevented the formation of the S fraction during heat shock. Overproduction of ClpB (exceeding by about eight-fold that of wild-type) in the same background did not prevent protein aggregation after heat shock and only partly compensated for the effect of the mutation in the clpB gene. Monitoring the S fraction during co production of DnaK/DnaJ/GrpE and ClpB in the DeltaclpB mutant revealed that both the levels of expression and the ratios of ClpB to Hsp70 system proteins had a significant effect on the formation and removal of protein aggregates in heat shocked E. coli cells. In the presence of excess ClpB, an increase in the levels of DnaK, DnaJ and GrpE was required to prevent aggregate formation upon heat shock or to efficiently remove protein aggregates after heat shock. Therefore, it is supposed that a high level of ClpB under some conditions, especially at insufficient levels of Hsp70 system proteins, may support protein aggregation resulting from heat shock and may lead to stabilization of hydrophobic aggregates. PMID- 11731149 TI - Development and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens. AB - Competitive PCR assays were developed for the enumeration of the rumen cellulolytic bacterial species: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens. The assays, targeting species-specific regions of 16S rDNA, were evaluated using DNA from pure culture and rumen digesta spiked with the relevant cellulolytic species. Minimum detection levels for F. succinogenes, R. albus and R. flavefaciens were 1-10 cells in pure culture and 10(3-4) cells per ml in mixed culture. The assays were reproducible and 11-13% inter- and intra assay variations were observed. Enumeration of the cellulolytic species in the rumen and alimentary tract of sheep found F. succinogenes dominant (10(7) per ml of rumen digesta) compared to the Ruminococcus spp. (10(4-6) per ml). The population size of the three species did not change after the proportion of dietary alfalfa hay was increased. All three species were detected in the rumen, omasum, caecum, colon and rectum. Numbers of the cellulolytic species at these sites varied within and between animals. PMID- 11731150 TI - Chitin degradation by cellulolytic anaerobes and facultative aerobes from soils and sediments. AB - Species of strictly and facultatively anaerobic cellulolytic bacteria from soils and sediments were examined for the ability to degrade chitin. Of 22 species studied, 16 degraded insoluble chitin. Cellulomonas uda, which was selected for a comparative study of its cellulase and chitinase enzyme systems, produced different enzyme systems for the degradation of cellulose and chitin and different patterns of regulation of production of the two enzyme systems were observed. Moreover, C. uda utilized chitin as a source of nitrogen for the degradation of cellulose. In natural environments, the ability to use chitin as a nitrogen source may confer on cellulolytic microorganisms, such as C. uda, a selective advantage over other cellulolytic microbes. PMID- 11731151 TI - Physiological difference during ethanol fermentation between calcium alginate immobilized Candida tropicalis and Saccharomyces cerevisiae. AB - Calcium alginate-immobilized Candida tropicalis and Saccharomyces cerevisiae are compared for glucose fermentation. Immobilized C. tropicalis cells showed a slight morphological alteration during ethanol production at 40 degrees C, but their fermentation capacity was reduced by 25%. Under immobilization conditions, the two species demonstrated two different mathematical patterns when the relationship between growth rate, respiration rate, and ethanol tolerance was assessed. The interspecific difference in behavior of immobilized yeast cells is mainly due to their natural metabolic preference. The production of CO(2) by calcium alginate-immobilized C. tropicalis, as well as the lower supply of oxygen to the cells, are the major factors that reduce ethanol production. PMID- 11731152 TI - Assimilatory nitrate reductase from the haloarchaeon Haloferax mediterranei: purification and characterisation. AB - Haloferax mediterranei can use nitrate as sole nitrogen source during aerobic growth. We report here the purification and biochemical characterisation of the assimilatory nitrate reductase (EC 1.6.6.2) from H. mediterranei. The enzyme, as isolated, was composed of two subunits (105+/-1.3 kDa and 50+/-1.3 kDa) and behaved as a dimer during gel filtration (132+/-6 kDa). A pH of 9 and elevated temperatures up to 80 degrees C (at 3.1 M NaCl) are necessary for optimum activity. The enzyme stability and activity of the enzyme depend upon the salt concentration. Reduced methyl viologen was as effective as the natural electron donor ferredoxin in the catalytic process. In contrast, NADPH and NADH, which are electron donors in nitrate reductases from different non-photosynthetic bacteria, were ineffective. PMID- 11731153 TI - Folic acid utilisation related to sulfa drug resistance in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae mutants deficient in folate synthesis have been constructed and employed to study the utilisation of exogenous folates in yeast. One mutant specifically lacked dihydropteroate synthase while the second lacked dihydrofolate synthase. Exogenous folinic acid restored optimal growth to both strains. Folic acid did not generally rescue growth but spontaneous isolates capable of utilising folic acid were selected. The folic acid synthesis pathway in the folate utilising isolates was restored via transformation with FOL1 or FOL3 expression plasmids and transformants were tested for resistance to sulfamethoxazole (SMX). The presence of elevated levels of folic acid led to greatly reduced SMX sensitivity regardless of whether strains were folate utilisers or not. PMID- 11731155 TI - Experimental inoculation of late term pregnant sows with a field isolate of porcine reproductive and respiratory syndrome vaccine-derived virus. AB - The use of a live attenuated porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in piglets has been associated with reproductive disorders in non vaccinated sows. Vaccine-derived virus (VDV) has been isolated from foetuses, stillborn pigs, and dead piglets, indicating that the live vaccine spread from vaccinated piglets to non-vaccinated sows, and that the virus might be implicated in the severe reproductive problems observed. In the present study, one such VDV isolate was used to experimentally infect pregnant sows in the last trimester. The chosen isolate, which had more than 99.6% identity to the attenuated vaccine virus, originated from the lungs of a stillborn pig from a swine herd with a sudden high level of stillborn pigs and increased piglet mortality in the nursing period. Intranasal inoculation of sows with the virus isolate resulted in congenital infection, foetal death, and preweaning pig mortality. As such, the present study showed that vaccine-derived PRRSV can cause disease in swine consistent with PRRS. PMID- 11731156 TI - Extensive antigenic and genetic variation among foot-and-mouth disease type A viruses isolated from the 1994 and 1995 foci in Sao Paulo, Brazil. AB - Nine foot-and-mouth disease virus (FMDV) type A isolates recovered from the field FMD foci in Sao Paulo State, Brazil, during 1994 and 1995 (a period preceding the last reported focus of FMD in 1996 in this state) were compared among themselves and with the reference vaccine strain A(24)Cruzeiro. The techniques used were sandwich ELISA, virus neutralization (VN), polyacrylamide gel electrophoresis (PAGE) of the structural polypeptides and direct sequencing of the VP1-coding region (1D gene). Results of VN were recorded as serological relationships "R" and those from ELISA were expressed as percentage of the homologous reaction "r". ELISA and VN gave comparable results (correlation coefficient, 0.936) allowing assignment of these field viruses to four groups which were distinct from the A(24)Cruzeiro strain. PAGE and 1D nucleotide sequencing were also able to distinguish between these viruses. The high level of genetic and antigenic variation found when comparing the A(24)Cruzeiro vaccine strain and type A strains recovered from the last identified foci of FMD came from a formerly endemic area where vaccination with polyvalent vaccines (O(1)Campos, A(24)Cruzeiro and C(3)Indaial) had been extensively applied. The similarity between the results of the serological and genetic analyses suggest that the antigenic differences found are mainly located in the 1D protein. PMID- 11731157 TI - Humoral immune responses following experimental infection of goats with Mycoplasma capricolum subsp. capripneumoniae. AB - Goats housed in microbiologically secure facilities were experimentally endobronchially infected with Mycoplasma capricolum subsp. capripneumoniae (Mccp), causal agent of contagious caprine pleuropneumonia (CCPP). The animals were monitored over an 8-week period post-infection (p.i.). Elevated temperatures were observed 2-7 days p.i., reaching a maximum of 41.5 degrees C in one animal (1884). By 8 weeks p.i. the infection was successfully cleared, with no Mccp being recovered from the lungs, serum or nasal passages. Mccp was not isolated from serum throughout the experiment, either directly by culture or indirectly via polymerase chain reaction (PCR). Humoral immune responses against Mccp capsular polysaccharide (CPS) were generally poor when measured by ELISA. CPS antigen was present in the serum of all infected animals early in the infection (day 14 p.i.), although in one animal (1855) CPS antigen persisted throughout. This was the only animal to exhibit a serious cough (day 5-19 p.i.). Successful diagnosis of CCPP was achieved using two different types of latex agglutination test (CPS antibody and CPS antigen detection test), immunoblotting and a blocking ELISA, although the latter lacked sensitivity until later in the infection (35-40 days p.i.). Only a single animal (1855) was detected positive using the current complement fixation test (CFT). Strong immune responses to protein antigens were detected by IgG and IgM immunoblotting from the first time point at day 14 p.i. IgM immunodominant bands of 220, 85, 62 and 40kDa were observed in the 3 infected animals and from CFT-positive CCPP field sera. Band intensity gradually diminished throughout the experiment. IgG immunodominant bands of 108, 70, 62, 44, 40 and 23kDa were shared between experimentally-infected and field sera, with band intensity either remaining unchanged or increasing from day 14 p.i. These bands were not present using pre-infection sera. Of the diagnostic tests used, only the CPS antibody detection latex agglutination test and IgG immunoblotting gave positive diagnoses throughout the entire period post-infection (days 14-53 p.i.). PMID- 11731158 TI - Proposal of a new serovar of Actinobacillus pleuropneumoniae: serovar 15. AB - We report on the re-examination of nine Australian isolates of Actinobacillus pleuropneumoniae that have been previously assigned to serovar 12. In the ring precipitation test, none of the nine isolates reacted with antisera to serovars 1 14 of A. pleuropneumoniae. Antiserum prepared against one of the Australian isolates gave no reaction with any of the 14 recognised serovar reference strains, except serovar 7. This reaction of the HS143 antiserum with serovar 7 antigen could be removed by adsorption with serovar 7 antigen. The adsorbed antiserum remained reactive with HS143 and the other eight Australian isolates. The nine Australian isolates were all shown to express ApxII and ApxIII, found in serovars 2, 4, 6 and 8, as well as the 42kDa outer membrane protein found in all serovars of A. pleuropneumoniae. The nine Australian isolates were found to possess the following toxin associated genes apxIBD, apxIICA, apxIIICA, apxIIIBD and apxIVA. The toxin gene profile of the Australian isolates is typical of A. pleuropneumoniae serovars 2, 4, 6 and 8. On the basis of the serological characterisation results and the toxin gene profiles, we propose that these isolates represent a new serovar of A. pleuropneumoniae--serovar 15--with HS143 being the reference strain for the new serovar. PMID- 11731159 TI - A putative iron-regulated TonB-dependent receptor of Mannheimia (Pasteurella) haemolytica A1: possible mechanism for phase variation. AB - A recombinant plasmid that codes for a novel iron receptor protein (Irp) of Mannheimia (Pasteurella) haemolytica A1 was isolated by the partial complementation of an Escherichia coli fur mutant. The deduced amino acid sequence of Irp exhibited characteristics typical of TonB-dependent receptors. These include: a TonB-box at the N-terminal; a 50 amino acid region homologous to the "plug" domain of the E. coli FhuA and FepA receptors; and a C-terminal TonB dependent signature which likely functions as an outer membrane anchoring domain. Previously uncharacterized Irp homologues were detected by BLAST analysis of available databases and incomplete microbial genomes. When the irp homologues from Neisseria gonorrhoeae and N. meningitidis were cloned by PCR and expressed in E. coli, novel proteins of the predicted size (84kDa) were detected in cell lysates, demonstrating that these are functional genes. The M. haemolytica A1 irp gene undergoes phase variation at a nucleotide region which contain the sequence AAAAAAATTAAAA (7A-2T-4A) flanked by a short inverted repeat. Site-specific mutagenesis of the 7A-2T-4A sequence as well as replacement of the inverted repeats resulted in a stable construct that expressed the Irp protein without phase variation. The expression of irp in M. haemolytica A1 was regulated by iron concentrations and most likely a Fur homologue, consistent with the proposed function of Irp in iron metabolism. The irp genes may represent contingency loci that play a role in iron acquisition during infection. PMID- 11731160 TI - Plasmid and restriction endonuclease patterns in Pasteurella multocida isolated from a swine pyramid. AB - Restriction endonuclease analysis (REA) and plasmid profile were used to study the epidemiology of Pasteurella multocida in a swine pyramid structure. The studied pyramid was comprised of a group of 12 swine farrow-to-finish farms related by unidirectional animal movement. P. multocida isolates were obtained from the lungs of 275 slaughtered pigs. Serotyping was performed by hyaluronidase sensitivity test and toxicity was investigated by the ELISA test. HpaII was used to cleave the P. multocida extracted DNA. REA patterns relationships were studied using the Sokal-Michener coefficients, and the dendrogram was built using the UPGMA system. The 218 P. multocida isolates obtained were distributed in 17 REA patterns. In 9 of the 12 farms studied only 2-3 REA patterns were detected, with one clearly predominant pattern. The 81 strains with plasmids were assigned to six plasmid profiles. REA and plasmid profiles proved to be good epidemiological tools for identifying different strains of P. multocida with the same phenotype. PMID- 11731161 TI - Salmonella enterica in reptiles of German and Austrian origin. AB - Captive reptiles are routinely identified as reservoirs of Salmonella spp. and the number of reports about reptile-associated salmonellosis is increasing. In the present study, Salmonella were detected in 86 of 159 (54.1%) faecal reptile samples cultured. The percentage of Salmonella positive samples was significantly lower in turtles as compared with lizards and snakes, as Salmonella were only detected in one sample from a single turtle out of 38 turtles investigated. In all, 42 different Salmonella serovars were found. All isolated Salmonella belonged to the species enterica, predominantly to the subspecies I (n=46) and IIIb (n=30), but also to subspecies II (n=3), IIIa (n=6) and IV (n=2). All isolates were sensitive to the antimicrobials examined. A comparison between the reptile owners indicated that either no Salmonella were found, or that Salmonella could be isolated from all or nearly all animals of the respective owners. A significantly higher percentage of Salmonella positive reptiles was detected in the group of owners who purchase reptiles in comparison with pure breeders. A total of 88.9% of Salmonella isolates were found in samples of reptiles bought in pet shops and 58.8% in samples from wild-caught animals. The high percentage of Salmonella in reptiles in our study confirms the risk for the transmission of the infection to humans. PMID- 11731162 TI - Identification of a new biotype of Actinomyces hyovaginalis in tissues of pigs during diagnostic bacteriological examination. AB - Bacterial strains isolated from a large variety of necropsy samples of pigs and previously described as a phenotypical homogeneous group were shown to belong to the species Actinomyces hyovaginalis. This was unexpected because their colonial characteristics, as well as their origins, were very different from those originally reported for the vaginal strains on which the species description of A. hyovaginalis was based. Colonial morphology, as well as fermentation of cellobiose, reactions in hippurate and nitrate and production of beta glucuronidase, allowed separation of the strains studied here from the vaginal strains. Analysis of tRNA intergenic length polymorphisms (tDNA-PCR), 16S rRNA gene sequencing and DNA-DNA hybridizations were carried out and led to the proposal of a separate biotype within the species A. hyovaginalis. Since, the strains were isolated from different body sites, this biotype has been designated as the 'general' biotype of A. hyovaginalis, while the strains on which the original species description was based are designated as the 'vaginal' biotype. PMID- 11731163 TI - Phenotypic and genotypic characterization of Mannheimia (Pasteurella) haemolytica like strains isolated from diseased animals in Denmark. AB - Trehalose-negative strains of the Pasteurella haemolytica complex have recently been transferred to a new genus, Mannheimia. This genus presently consists of five named species: M. haemolytica, M. glucosida, M. granulomatis, M. ruminalis and M. varigena. The purpose of this study was to investigate the occurrence of these species and lesions associated with these isolates in Denmark. In all 106 M. haemolytica-like strains isolated from pathological material from cattle, sheep, pigs and hares submitted to the Danish Veterinary Laboratory between 1994 and 1998 were investigated. Phenotypic characterization and ribotyping were used for identification in addition to sequencing of the 16S rRNA genes for selected strains. The species allocation was determined by comparison to results from a previous polyphasic taxonomic study. Seventy-one percent of the strains belonged to M. haemolytica, 18% to M. varigena and 8% to unnamed groups within the genus Mannheimia. Single isolates identified as M. glucosida and P. trehalosi, respectively, were detected. Two isolates belonged to M. granulomatis. Forty three percent of the strains belonged to serotype 1, 41% were untypeable, while the rest belonged to serotypes 2, 7, 9, and 16. The present investigation also showed that a simplified phenotypic characterization using Diatabs Diagnostic Tablets (Rosco, Denmark) represents a useful method for obtaining a quick and reliable species identification. Finally, the investigation confirmed that serotyping does not represent a reliable method for species identification. The heterogeneity of species associated with bovine "pasteurellosis" should be considered in future studies to improve our understanding of the pathogenesis of pneumonic disease. PMID- 11731164 TI - Are goats naturally resistant to gastric Helicobacter infection? AB - Gastric Helicobacter species are widespread and have been reported in wild and domestic mammals of different dietary habits such as humans, dogs, cats, macaques, mice, cheetahs, ferrets, swine and cattle. All have been associated with gastric pathologies. Recently, gastric Helicobacter species were shown to be widespread in cattle and swine in Europe, and there is a report of Helicobacter pylori in sheep in Italy. However, there are no reports of Helicobacter infection in the goat, another important domestic animal of human consumption. The aim of our study was to assess whether Helicobacter abomasal infection was common in goats slaughtered for human consumption. Infection was detected through PCR analysis of DNA extracted from gastric biopsies, using genus- and species specific primers. Bovine and porcine gastric samples were also analyzed as positive controls. None of the 70 goats were positive for Helicobacter spp.; however, Candidatus Helicobacter bovis and Candidatus Helicobacter suis were detected in 85% of the bovine and 45% of the porcine samples, respectively. We discuss the possibility that goats may exhibit natural resistance to abomasal infection by Helicobacter spp. PMID- 11731165 TI - Antimicrobial susceptibility testing of Australian isolates of Brachyspira hyodysenteriae using a new broth dilution method. AB - The antimicrobial susceptibilities of 76 field isolates of Brachyspira hyodysenteriae from different states of Australia were tested in a newly developed broth dilution procedure. The antimicrobial agents used were tiamulin, valnemulin, tylosin, erythromycin, lincomycin and clindamycin. The results from the broth dilution susceptibility testing of 39 of the isolates were compared with results obtained for the same isolates using the agar dilution method. Amongst the isolates tested by broth dilution, 17 were from three farms and had been collected over a number of years. Their pulsed field gel electrophoresis pattern previously had been determined. The broth dilution technique was simple to use, less labor intensive than agar dilution, and gave clear end points. The results obtained using the two methods generally corresponded well, although in a few cases the MIC obtained by broth dilution were lower than those with agar dilution. For the 76 isolates tested by broth dilution, the MIC(90) (mg/l) was: tiamulin, 1; valnemulin, 0.5; tylosin>256; erythromycin>256; lincomycin, 64 and clindamycin, 16. Only minor differences in susceptibility patterns were found amongst isolates from different Australian states. Over all the isolates, and also amongst the isolates obtained from different years on the three farms, there was no trend for the susceptibility of the isolates to alter with time. PMID- 11731166 TI - Genetic and enzymatic analyses of metalloprotease (aureolysin) from Staphylococcus aureus isolated from domestic animals. AB - The gene (aur) encoding the metalloprotease (aureolysin) of Staphylococcus aureus from domestic animals was analyzed by polymerase chain reaction (PCR), PCR restriction fragment length polymorphism (PCR-RFLP) and sequencing. The aur gene was detected in all 74 isolates from cows, pigs and chickens by PCR amplification and was classified into types I and II by PCR-RFLP patterns. The type II aur gene was contained in 36 (94.7%) of 38 protease-positive isolates as judged by skim milk agar plate culture, while type I was contained in 28 (77.8%) of 36 protease negative isolates. The deduced amino acid sequences of aureolysins from type I and II isolates were almost identical with those of the published data. Subsequently, the two aureolysins were purified from the culture supernatants of type I and II isolates. The molecular weights of purified type I and II aureolysins were both estimated at 34kDa by SDS-polyacrylamide gel electrophoresis. These aureolysins had maximum proteolytic activity at 30-50 degrees C and pH 7.0-8.0. Their activity was inhibited by metal- and zinc specific inhibitors, such as EDTA, EGTA and 1,10-phenanthroline. Specific activity (activity/protein) of type II aureolysin was two times higher than that of type I. These results indicated that the aur gene is highly conserved with two allelic forms (types I and II) among bovine, porcine and avian isolates of S. aureus. PMID- 11731167 TI - Escherichia coli strains isolated from the uterus and urinary bladder of bitches suffering from pyometra: comparison by restriction enzyme digestion and pulsed field gel electrophoresis. AB - Pyometra (uterine inflammation with accumulation of pus in the uterus) is regarded as one of the most common illnesses in bitches. The ethiology and pathogenesis are complex with both hormonal and bacterial elements. The bacteria most frequently isolated from the uterine content is Escherichia coli. In this study, 84 E. coli strains from the uteri of 70 bitches suffering from the disease were examined and their DNA-profiles compared by restriction enzyme analysis and pulsed-field gel electrophoresis (PFGE). Through variations in DNA-profiles of the E. coli isolates, this study indicates that pyometra is caused by E. coli originating from the normal flora of the dogs and not by certain clones spread between animals.E. coli strains from the urinary bladder and the uterus of six of the bitches suffering from simultaneous urinary tract infection and pyometra were examined and compared as above. The DNA-profiles of the isolates from each of the six bitches were 100% identical. This study supports the theory suggesting that in cases of simultaneous urinary tract infection and E. coli pyometra, the urinary tract and uterus are infected with the same bacterial strain. To evaluate whether the uterus was infected with a single clone of E. coli or if multiple clones were present, eight to 16 colonies of E. coli isolated from pyometra samples from a further 10 bitches were examined. All bacterial colonies from the culture of the same bitch showed identical DNA-profiles. In 14 of the 70 bitches, two macroscopically different but biochemically identical E. coli colony types were isolated. The two colony types from the same bitch proved to have identical DNA-profiles in 13 cases and almost identical in the remaining bitch. PMID- 11731168 TI - Assessment of protective efficacy of live and killed vaccines based on a non encapsulated mutant of Streptococcus suis serotype 2. AB - The protective efficacy of a live and killed non-encapsulated isogenic mutant of Streptococcus suis serotype 2 was determined in pigs, and compared with the efficacy of the capsulated wild-type strain. SPF pigs were vaccinated twice intramuscularly at 4 and 7 weeks of age with a dose of 1 x 10(9) formalin-killed CFU of the wild-type (WT-BAC), formalin-killed non-encapsulated mutant (CM-BAC) or live non-encapsulated mutant (CM-LIVE) strain. After 2 weeks, vaccinated pigs and non-vaccinated controls were challenged intravenously with 1 x 10(7) CFU of the homologous, wild-type S. suis serotype 2 strain. Protection was evaluated by clinical, bacteriological, serological and post-mortem examinations. All pigs vaccinated with WT-BAC were completely protected against challenge with the homologous serotype. Pigs vaccinated with CM-BAC were partially protected. Although all pigs vaccinated with CM-BAC survived the challenge, four out of five pigs developed clinical signs of disease for several days. Compared to the WT-BAC and CM-BAC, the CM-LIVE vaccine was less protective. Two out of five pigs vaccinated with CM-LIVE died in the course of the experiment and all of them developed specific clinical signs of disease for several days. The protective efficacy of the vaccines could be associated with serum antibody titers. Antibody titers against cells of wild-type and non-encapsulated mutant strains as well as against muramidase-released proteins (MRP) were high in pigs vaccinated with WT BAC and CM-BAC. Pigs vaccinated with CM-LIVE showed lower antibody titers. Antibody titers against purified capsular polysaccharides (CPS) of S. suis serotype 2 were only found in pigs vaccinated with WT-BAC. These findings indicate that CPS and other bacterial components of WT-BAC are probably essential for full protection against homologous challenge. PMID- 11731169 TI - Antimicrobial susceptibility and plasmid analysis of Actinobacillus pleuropneumoniae isolated in Taiwan. AB - Sixty Actinobacillus pleuropneumoniae (App) strains from pigs in Taiwan were examined. Serotyping revealed that these belonged to serovars 1 (n=53), 2 (n=3), and 5 (n=4). Agar disk diffusion susceptibility testing of the isolates showed 55 (92%) were resistant to three or more antimicrobial agents. Six resistance patterns were observed. Ampicillin-chloramphenicol-flumequine-nalidixic acid streptomycin-sulfonamide/trimethoprim-tetracycline was the most common multi resistance pattern. Minimal inhibitory concentration of 14 antimicrobial agents was determined. The isolates were highly susceptible to ceftiofur and trimethoprim in vitro. Isolates were resistant to streptomycin, ampicillin, and nalidixic acid. All isolates were examined for the presence of plasmids using the alkaline lysis method. Forty three (72%) isolates had four plasmid bands with an approximate sizes of 3.5, 4.3, 5.8 and 6.0 kb; 12 (20%) had three bands at 3.5, 4.3 and 5.2 kb, and 5 (8%) had no plasmid bands. Antimicrobial resistance plasmids were detected in resistant strains of App. Three antimicrobial resistance plasmids were transformed into E. coli DH5 alpha. pTMY1 (4.3 kb) encoded a streptomycin kinase and a dihydropteroate synthase; pTMY2 (6.0 kb) encoded ROB-1 beta-lactamase and aminoglycoside 3'-phosphotransferase; pTMY3 (5.2 kb) encoded only ROB-1 beta-lactamase. The 4.3 kb plasmid was sequenced and consisted of 4242 bp with 42.9% GC content. The 4.3 kb plasmid DNA sequence was 98% homologous to a plasmid previously isolated from Pasteurella haemolytica. PMID- 11731170 TI - Characterization of turkey coronavirus from turkey poults with acute enteritis. AB - The present study was to characterize turkey coronavirus associated with turkey poult enteritis and mortality. Intestinal contents or intestines from affected turkey poults and inoculated turkey embryos contained coronaviruses as revealed by electron microscopy or were positive for turkey coronavirus by immunofluorescent antibody assay. Sucrose density gradient ultracentrifugation of the virus-containing intestinal homogenate yielded two opalescent bands corresponding to the buoyant densities of 1.14-1.15 and 1.18-1.20 g/ml, respectively. Coronaviral particles from intestinal contents or the sucrose density gradient preparation were mainly spherical in shape and had envelope and central depression. They were surrounded by a fringe of regularly spaced petal shaped projections attached to the particles by a short stalk. Purified viruses hemagglutinated rabbit erythrocytes with a titer of 16. Major protein bands of purified viruses analyzed by SDS-PAGE were located at 200, 100-110, 50-60, and 30 35 kDa. The patterns of protein bands were consistent with those of Minnesota or Quebec turkey coronavirus isolates. A 568 bp nucleotide fragment of turkey coronavirus spike protein gene was amplified from RNA of inoculated turkey embryo intestine or purified virus. Sequence analysis of the 568 bp PCR product revealed high degree of identity with the corresponding spike protein gene sequence of human and bovine coronaviruses. The results indicated that turkey coronavirus was associated with turkey poults with acute enteritis. PMID- 11731171 TI - Isolation and characterization of an imipenem-resistant Bacteroides fragilis strain from a prostate abscess in a dog. PMID- 11731172 TI - Serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP) with an indirect ELISA based on the specific lipoprotein LppQ of Mycoplasma mycoides subsp. mycoides SC. AB - An indirect ELISA, based on the specific and strongly antigenic recombinant peptide of the N'-terminal half of the lipoprotein LppQ from Mycoplasma mycoides subsp. mycoides small colony type (SC) was developed for the detection of antibodies to M. mycoides subsp. mycoides SC. It was evaluated for its suitability for serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP). The recombinant peptide containing poly-histidine residue tails was expressed in Escherichia coli and subsequently purified by Ni(2+) chelate affinity chromatography to be used as antigen to coat microtiter ELISA plates. The specificity of the antigen was tested against rabbit hyperimmune sera directed against related Mycoplasmas of the M. mycoides cluster and with sera from cattle that were either free of CBPP, but suffered from other mycoplasmal infections such as M. bovis, or showed cross-reactions in the complement fixation test. The sensitivity of the ELISA was assessed with sera from artificially infected animals and with sera from cattle originating from areas where CBPP was endemic at the time of blood sampling. The study revealed that the ELISA was both specific and sensitive for CBPP positive bovine sera and was shown also to be robust to harsh climatic conditions. PMID- 11731173 TI - The effect of dietary spray-dried porcine plasma on clinical response in weaned piglets challenged with a pathogenic Escherichia coli. AB - Weaned piglets were used to determine the effect of dietary spray-dried porcine plasma (SDPP) on the clinical response to an infection with a pathogenic Escherichia coli (E. coli) O139:K82 LT(-). The piglets were divided into two groups of 10 animals each. One group was fed the control diet containing soybean(meal) plus whey powder. The test piglets were fed a diet with 8% SDPP. Piglets were orally infected with the challenge strain on days 6 and 7 after weaning. The experimental period lasted 14 days after which the piglets were euthanised and necropsied. Faecal samples were collected daily for bacteriological analysis. Segments of jejunum, caecum and rectum were removed for bacteriological analysis post mortem. Feed intake and weight gain, faecal and condition scores and body temperature were measured daily. In the control and SDPP groups, 6 and 7 piglets died from diarrhoea. The average daily feed intake (ADFI) and average daily gain (ADG) were substantially higher in the SDPP group than in the control group. SDPP-fed piglets generally had a more favourable faecal score and a healthier appearance than did the control piglets. The faecal excretion of E. coli O139:K82 was similar for control and test piglets. There were no diet effects on the E. coli O139:K82 counts at different sites of the intestine. In this experiment, the inclusion of SDPP at an economically acceptable percentage in the diet could not prevent piglet losses due to challenge with a pathogenic E. coli, but improvements of ADG, ADFI and faecal and condition scores were achieved. PMID- 11731175 TI - Examination of Escherichia coli from poultry for selected adhesin genes important in disease caused by mammalian pathogenic E. coli. AB - A collection of 1601 extraintestinal and intestinal Escherichia coli isolated from chickens, turkeys and ducks, in Belgium, France and Spain, was hybridised with gene probes specific for fimbrial and afimbrial adhesins (F17, F18, S , Bfp, Afa, Cs31A, Intimin , Aida-1) of intestinal, urinary and invasive E. coli of mammals and with a probe specific for the P (Pap/Prs) fimbrial adhesin of urinary and invasive E. coli of mammals and birds. Three hundred and eighty-three strains (23.9%) were P-positive, 76 strains (4.8%) were Afa-positive, 75 strains (4.7%) were F17-positive, 67 strains (4.2%) were S positive, 23 (1.4%) were Intimin-positive, and all were F18-, Cs31A-, Aida1- and Bfp-negative. The 75 F17-positive strains harboured different major subunit A encoding gene variants, but the f17Ac variant was the most frequent (52 strains, 69.3%) and seven strains (9.3%) were not typeable. The f17G gene variant coding for the GII adhesin was the most frequent (56 strains, 75.0%), whereas the f17GI gene variant was present in four strains (5%) and 15 strains (20.0%) were not typeable. All Afa-positive strains harboured the afa-8 variant. The 23 Intimin positive E. coli tested positive for the beta-variant (16 strains; 69.6%) or for the gamma-variant (seven strains; 30.4%) of the eae gene. Chicken and turkey E. coli were more frequently probe-positive (43.6 and 43.1%, respectively) than duck E. coli (31.5%) and extraintestinal E. coli were also more frequently probe positive (48.4%) than intestinal strains (18.5%). Different combinations of probe positive hybridisation results were observed in 72 of the 540 probe-positive E. coli (13.3%). The most frequent combinations were between AfaE-8 and F17 probes (47 strains; 8.7%) and between P and S probes (13 strains; 2.4%). Although f17- and afa-8-related DNA sequences can be plasmid-located in mammalian E. coli, they were not in avian E. coli. Besides the P fimbrial adhesins, F17 and S fimbrial and Afa-VIII and Intimin afimbrial adhesins may thus represent colonisation factors of avian pathogenic E. coli. PMID- 11731174 TI - Vaccination of ducks with recombinant outer membrane protein (OmpA) and a 41 kDa partial protein (P45N') of Riemerella anatipestifer. AB - The generation of protective immunity against Riemerella anatipestifer infection in ducks were investigated by immunizations with recombinant glutathione sulfatransferase (GST) fusion's proteins of OmpA, a 42kDa major outer membrane protein, and P45N', a 41kDa N-terminal fragment of a newly identified 45kDa potential surface protein from R. anatipestifer. The DNA encoding OmpA and P45N' were isolated from R. anatipestifer serotype 15 (field strain 110/89) and serotype 19 (reference strain 30/90), respectively. Immunoblotting and ELISA results showed that the purified recombinant proteins induced the production of antibodies in immunized ducks. However, neither was protective against subsequent challenge with the virulent serotype 15 strain, 34/90. All the five ducks immunized with formalinized R. anatipestifer strain 34/90 survived the challenge with the homologous strain whereas six out of seven ducks in the non-immunized control group died within a week following the challenge. PMID- 11731176 TI - Antigen-induced production of interferon-gamma in samples of peripheral lymph nodes from sheep experimentally inoculated with Mycobacterium avium subsp. paratuberculosis. AB - The production of interferon-gamma (IFN-gamma) in response to Johnin purified protein derivate was measured in samples of the prescapular lymph node (PLN) from 10 sheep, aged 2 years, and nine sheep, aged 1 year that had been inoculated orally with Mycobacterium avium subsp. paratuberculosis within their first month of life. Ten non-inoculated sheep, aged 1 year, constituted the negative control group. The results obtained in the PLN IFN-gamma assay were compared with those derived from serological tests: a complement fixation test (CFT), agar gel diffusion test (AGID) and enzyme-linked immunosorbent assay (ELISA), as well as an IFN-gamma test on samples of blood. Among the 19 inoculated sheep, 16 gave positive reactions in the PLN IFN-gamma assay on samples incubated overnight, and 18 tested positive when the assay was applied to PLN samples incubated for 48h. In comparison, three, four and seven inoculated sheep gave positive reactions in the ELISA, CFT and in the blood IFN-gamma assay on samples incubated overnight, respectively. The AGID and IFN-gamma assay on blood samples incubated for 48h detected eight inoculated animals. Twelve inoculated sheep, that tested positive in the PLN IFN-gamma assay were clinically normal, gave negative results in an IS900-based polymerase chain reaction (PCR) assay on samples of ileum and ileocaecal lymph node and had no histological evidence of paratuberculosis, but tested positive on more than two occasions in sequential serological testing before necropsy. None of the 10 non-inoculated sheep tested positive in the AGID, CFT, ELISA, blood IFN-gamma assay on samples incubated overnight and for 48h or the PLN IFN-gamma assay on samples incubated overnight, but one gave a positive result in the PLN IFN-gamma assay on samples stimulated for 48h. It is likely that the positive reactions obtained by the PLN IFN-gamma assay in the 12 inoculated sheep that tested negative in the PCR assay and histopathological examination represents immunological evidence of latent infection or previous exposure to M. paratuberculosis rather than active infection. PMID- 11731177 TI - Development of a Brucella suis specific hybridisation probe and PCR which distinguishes B. suis from Brucella abortus. AB - A genomic library was prepared from Brucella suis DNA (MboI digested) and cloned into the BamHI site of pUC18. Colony hybridisation using a probe prepared from purified B. suis DNA labelled with alpha 32P was carried out to identify colonies of interest. About 20 colonies, which gave an intense signal upon hybridisation with whole B. suis genomic DNA as a probe, were selected. Because of the high degree of DNA homology between B. suis and Brucella abortus, a short probe was chosen as it would more likely give species specificity. Of seven fragments selected to probe whole B. suis, B. abortus, and Yersinia enterocolitica DNA, one was found to hybridise with B. suis only. The probe was sequenced in two directions and sense and anti sense primers of 25bp in length were chosen to yield a product of 421bp. After optimisation of the PCR, a product of 420bp was obtained with B. suis template DNA and two bands of 420 and 650bp were detected with B. abortus template DNA. This is the first reported PCR of the Brucella genome where a single pair of primers will discriminate between B. suis and B. abortus. No band was observed when the two primers were used to amplify E. coli, Y. enterocolitica, Enterobacter cloacae, Staphylococcus aureus, Streptococcus uberis, Corynebacterium bovis, or Serratia marcescens template DNA. PMID- 11731178 TI - Attenuation of Actinobacillus pleuropneumoniae by inactivation of aroQ. AB - Actinobacillus pleuropneumoniae is the aetiological agent of porcine pleuropneumonia, a disease resulting in morbidity and mortality of pigs and accordingly economic losses within the swine industry. In order to construct a potential vaccine strain of A. pleuropneumoniae for control of this disease, the aroQ gene, required for the aromatic biosynthetic pathway, was targeted for inactivation. The resulting strain was tested for virulence within pigs. The aroQ gene and an adjacent gene, dapD, were cloned. A recombination cassette, for inactivation of aroQ, was constructed from these cloned genes by inserting an ampicillin resistance gene and this was transformed into A. pleuropneumoniae. Integration of this construct into the chromosomal location of aroQ and disruption of the aroQ/dapD gene arrangement was confirmed through PCR and Southern analysis. The resulting HS25 aroQ mutants were unable to grow in a chemically defined medium and following intratracheal delivery to pigs were only able to induce lung lesions when given at a level 10,000 times greater than that of the parent strain. Complementation with an in trans, functional, aroQ gene restored the ability of the mutant strain to grow in a chemically defined medium and virulence, when tested in pigs, confirming attenuation results from inactivation of aroQ. In conclusion, this work has constructed a defined mutant of A. pleuropneumoniae that is attenuated and may be safely delivered live to pigs. PMID- 11731180 TI - Hormone replacement therapy: the benefits in tailoring the regimen and dose. AB - Despite the clear benefits of long-term hormone replacement therapy (HRT), the majority of patients tend to undergo short-term treatment. The cyclical bleedings induced by the sequential progestogen administration are often unacceptable namely in the elderly postmenopausal women. At the standard doses HRT preparations can also induce annoying hormone-related side effects, both in sequential and continuous combined regimens. Lower HRT schedules are reported to be highly effective in the relief of climacteric symptoms, inducing minimal endometrial stimulation with high rates of amenorrhea. Continuous administration of low doses of progestins is safe for endometrium protection and minimizes progestin-related side effects. Indeed, it has been demonstrated that low dose HRT can prevent the increase in bone turnover and the consequent bone loss in postmenopausal women. The choice of lower HRT dosages can also be useful for the number of potential disadvantages of standard HRT doses, mainly for long-term treatments. Low dose regimens should be considered as a starting dose to minimize the occurrence of side effects, improving compliance and, therefore, HRT effects on the prevention of long-term consequences of estrogen deprivation. PMID- 11731179 TI - Vertical transmission of bovine leukemia virus and bovine immunodeficiency virus in dairy cattle herds. AB - Vertical transmission of bovine leukemia virus (BLV) and bovine immunodeficiency virus (BIV) was investigated in five dairy cattle herds in Hokkaido, where 36.1 and 17.0% of cattle were BLV and BIV seropositive, respectively, and 9.9% of dams were co-infected with both BIV and BLV. Twenty six cases of offspring born from dams infected with only BLV (17 cases) or with both BIV and BLV (9 cases) were examined for the presence of BLV and BIV before and after colostrum feeding by polymerase chain reaction (PCR) and syncytium assay. After birth, all calves were separated immediately from their dams. The offspring born from BLV-positive dams were BLV-negative before colostrum feeding, suggesting that no transplacental transmission had occurred. Thereafter, these offspring were fed colostrum or milk from their dams, but still remained BLV-negative. The other offspring born from BLV-positive dams were fed with BLV-negative colostrum, or with pasteurized BLV positive colostrum. All these calves remained negative for BLV infection, suggesting that in utero transmission of BLV is negligible. In the case of offspring born from dams co-infected with BLV and BIV, calves were BIV-positive before colostrum feeding at 1 day after the birth, indicating in utero transmission of BIV. After colostrum feeding from their dams, newborn calves became BLV-positive. In addition, one calf was BLV-positive even before colostrum feeding. These results suggest that BIV can be transmitted to offspring in utero, and that BLV can be transmitted through colostrum or milk if dams are infected with both BIV and BLV. PMID- 11731181 TI - Comparison of pharmacokinetic profiles of a 17 beta-estradiol gel 0.6 mg/g (Gelestra) with a transdermal delivery system (Estraderm TTS 50) in postmenopausal women at steady state. AB - OBJECTIVES: to compare the patterns of a 17 beta-estradiol (E(2)) gel containing 0.6 mg/g (1.5 mg E(2) per day, Gelestra); with the transdermal delivery system (Estraderm TTS 50) applied every 3 days over a 14-day period to women in spontaneous or surgical menopause. METHODS: a single centre, open, randomised, parallel-group study was conducted. A total number of 33 postmenopausal women were enrolled. In 23 of them the menopause occurred spontaneously, while 10 women were bilaterally ovariectomized. Randomly, the subjects were treated with Estraderm TTS 50 (no. 8) or with Gelestra (no. 14). The pharmacokinetic study of the drugs was performed at the seventh, ninth and 14th day in Gelestra treated women and at the first, third and second day in Estraderm TTS 50 treated women. In fact, the seventh, ninth and 14th day of percutaneous treatment corresponds to the first, third and second day of application of the transdermal system application. Blood samples were taken by each subject at baseline and 1, 2, 3, 4, 8, 12 and 24 h after the gel or transdermal system application. In almost all samples the level of E(2) and estrone (E(1)) were evaluated. Statistical analysis was performed by comparing the two groups of treatment. The following parameters were assessed: mean E(2) and E(1) concentrations, E(2) peak serum concentration within interval from 0 to 72 h (C(max)), E(2) trough concentration within interval from 0 to 72 h (C(min)), area under the E(2) time concentration curve in the interval from 0 to 72 h (AUC((0-72))), the average E(2) concentration during the measurement interval, calculated by dividing AUC((0-72)) by 72 h (C(av)), E(1)/E(2) ratio, and percentage fluctuation (%Fluct) which is equal to 100 (C(max)-C(min)/C(max)). RESULTS: there was no significant difference in E(2) C(av) between the two treatments. However, significant differences in favour to the gel on the first day (first h) and on third day (72nd h) and in favour to the patch at the second day (48th h) were detected. C(max), E(1)/E(2) ratio and AUC((0-72)) were not statistically different, while a significantly higher C(min) for the gel was observed. Furthermore, the 90% confidence interval for AUC((0 72)) ratio (0.83-1.10) was within the commonly applied bioequivalence acceptance range (0.80-1.25). The %Fluct was significantly lower for Gelestra than for Estraderm TTS 50. CONCLUSIONS: although the mean E(2) and E(1)concentrations, C(max), E(1)/E(2) ratio and the AUC((0-72)) did not differ between the two E(2) treatments, the Gelestra treatment showed a lower day-to-day variation over the three day application, than the Estraderm TTS 50. PMID- 11731182 TI - Evaluation of methods for prediction of bone mineral density by clinical and biochemical variables in perimenopausal women. AB - OBJECTIVES: to predict spinal and femoral bone mineral density (BMD) in perimenopausal women from simple clinical and biochemical variables. METHODS: 2016 women 3-24 months past last menstrual bleeding. Mean age 50.1+/-2.8 years. Age, height, weight, number of full term pregnancies, weekly hours of physical activity, sunbathing habits, use of sun bed, daily intake of calcium and vitamin D, smoking habits, consumption of alcohol, coffee, and tea, history of forearm or femoral neck fractures among the parents, serum osteocalcin (S-OC), serum bone specific isoenzyme of alkaline phosphatase (BSAP), and urine hydroxyproline/creatinine ratio (U-OHP) were used as predictors in three different mathematical models. Lumbar spine (L2-L4) and femoral neck BMD were measured by DEXA. Three mathematical models (multiple regression, logistic regression, and discriminant analysis) were applied. RESULTS: the multiple regression explained 19-21% of the total variation, and the logistic regression and discriminant function had a sensitivity between 53 and 67% with specificity ranging from 67 to 80%. Age, S-OC, serum bone specific alkaline phosphatase, and a maternal history of forearm or femoral neck fractures seemed to be reproducible risk factors for low bone mineral density irrespective of the mathematical model applied. When applied to a separate population, the models performed poorly. CONCLUSIONS: Simple clinical and biochemical variables are not useful to predict spinal and femoral BMD in the individual perimenopausal woman. PMID- 11731183 TI - Comparative effect of hormone replacement therapy on bone mass density and skin tensile properties. AB - OBJECTIVES: connective tissues constitutive of skin and bones are affected during the climacteric. Hormone replacement therapy (HRT) can help mitigate their atrophy. The aim of this study was to compare the HRT effect on the skin tensile properties and bone mass density. METHODS: a total of 120 postmenopausal women (60 untreated, 60 receiving HRT) were enrolled in the study. Skin tensile properties were assessed on the volar forearm using a computerized suction device. A 500 mbar suction was applied through a 4-mm diameter hollow probe. Two operating modes were applied using a steep and a progressive suction, respectively. BMD was measured on the hip, femoral neck and lumbar spine using dual X-ray absorptiometry. RESULTS: in both groups of women skin elasticity was correlated with BMD. HRT significantly reduced the climacteric-associated decline in skin elasticity. A trend in better preserved BMD was also found in these women without, however, reaching significance. CONCLUSIONS: it is concluded that measures of the skin tensile properties can be sensitive enough to disclose HRT efficacy upon connective tissue atrophy. Any decrease in skin elasticity during the climacteric should prompt to perform a BMD assessment. PMID- 11731184 TI - Effects of the combined use of calcitonin and 1 alpha-hydroxycholecalciferol on vertebral bone loss and bone turnover in women with postmenopausal osteopenia and osteoporosis: a prospective study of long-term and continuous administration with low dose calcitonin. AB - OBJECTIVES: The present study investigates the effect of long-term and continuous treatment with low dose calcitonin in combination with 1 alpha hydroxycholecalciferol on vertebral bone mass in early postmenopausal women. METHODS: A total of 202 postmenopausal women between 53 and 58 years of age were recruited individually and randomly assigned to one of four groups. Comparisons were made among groups of women receiving calcitonin alone (10 IU i.m. twice a month), 1 alpha-hydroxycholecalciferol alone (0.5 microg orally twice daily), a combination of the above two agents, or no treatment. Bone mineral density (BMD) of lumber spine (L2-4) was determined using Dual Energy X-ray Absorptiometry. The study was carried out prospectively over a 2-year period. RESULTS: We observed a significant increase in vertebral bone mass in the combined treatment regimen of calcitonin and 1 alpha-hydroxycholecalciferol (3.44% at 12 months in the combination group vs 1.40,0.92, and -0.70% in the calcitonin alone, 1 alpha hydroxycholecalciferol alone, and control groups, respectively; 4.51% at 24 months in the combination group vs 2.21,1.04, and -3.61% in the calcitonin alone, 1 alpha-hydroxycholecalciferol alone, and control groups, respectively). Serum PTH, osteocalcin levels and alkaline phosphatase activity decreased significantly within 12 months whereas urinary pyridinoline/creatinine ratio decreased at 24 months in the combination group. We observed mild adverse effects in 25.0% (7/28) and 30.0% (6/20) of combination regimen and calcitonin treatment cases, respectively. CONCLUSIONS: The results of the study suggest that the combination treatment regimen increased vertebral bone loss in early postmenopausal women to a greater extent than did calcitonin alone or 1 alpha-hydroxycholecalciferol alone. PMID- 11731185 TI - Effect of estradiol valerate alone or in association with cyproterone acetate upon vascular function of postmenopausal women at increased risk for cardiovascular disease. AB - OBJECTIVES: a large body of evidence has been accumulated suggesting that impairment of vascular endothelial function is an initial step in the development of atherosclerosis. Recent studies have shown that estrogen replacement therapy in postmenopausal women (PMW) improves endothelium-dependent, flow-mediated dilatation (FMD) while the cyclical adjunct of a progestin may reverse this effect. METHODS: the purpose of this study was to evaluate endothelium-dependent, FMD in the brachial artery and the plasma levels of Endothelin-1 in menopausal females treated with estradiol valerate with and without cyclical cyproterone acetate in 20 PMW (mean age 64+/-6 years) with more than one risk factor for coronary artery disease. After a baseline evaluation, PMW entered a double blinded, placebo controlled single cross-over study and were randomized to receive either estradiol valerate (2 mg) for 21 days or estradiol valerate (2 mg) for 11 days and estradiol valerate (2 mg) and cyproterone acetate (1 mg) for 10 days. Patients were crossed-over the complementary treatment 7 days after completing the first treatment phase. The study of forearm blood flow was repeated at the end of each treatment period. RESULTS: estradiol valerate significantly increased FMD as compared with baseline (12+/-3 vs. 7+/-4%, P<0.01) the adjunct of cyproterone acetate did not affect the effect of estradiol valerate upon FMD (12+/-3 vs. 11+/-4%, P=NS). Similarly reactive hyperemic flow increased after estradiol valerate alone (24%) or in association with cyproterone acetate (24%) compared with baseline. Plasma levels of Endothelin-1 were significantly reduced by estradiol valerate alone or in association with cyproterone acetate. CONCLUSIONS: in conclusion hormone replacement therapy with estradiol valerate and cyproterone acetate improves endothelial function and reduces plasma levels of Endothelin-1 in PMW at risk of coronary artery disease. These effects may be relevant for cardioprotection. PMID- 11731187 TI - Effective bleeding control and symptom relief by lower dose regimens of continuous combined hormone replacement therapy: a randomized comparative dose ranging study. AB - OBJECTIVES: We compared two different continuous combined hormone replacement therapy (HRT) regimens of estradiol valerate (E(2)V) and medroxyprogesterone acetate (MPA) with a combination of micronized estradiol (E(2)) and norethisterone acetate (NETA) to determine bleeding pattern, control of climacteric symptoms, lipid profile, endometrial and general safety in a 1-year multicenter study. METHODS: 440 postmenopausal women were randomized to three treatment groups to receive: 1 mg E(2)V+2.5 mg MPA; 1 mg E(2)V+5 mg MPA; or 2 mg of E(2)+1 mg NETA. After the first 6 months, the E(2)V dose was increased to 2 mg in both E(2)V/MPA groups. Information on bleeding was recorded on diaries by the women and intensity of climacteric symptoms was assessed using VAS scales. Physical and laboratory examinations, endometrial biopsy and vaginal ultrasonography were performed at baseline and follow-up visits. RESULTS: Significantly fewer bleeding days were experienced in the first 3 months by women taking E(2)V/MPA compared with women taking E(2)/NETA. When the dose of E(2)V was increased in the E(2)V/MPA groups, an increase in maximum bleeding intensity was observed in the group receiving 2.5 mg of MPA, but not in the group taking 5 mg of MPA. All dose combinations effectively relieved climacteric symptoms and beneficial effects on the lipid profile were seen after 6 months in all groups. Tolerability and endometrial safety were good and no cases of hyperplasia were observed. More women discontinued treatment prematurely in the E(2)/NETA group compared with either of the E(2)V/MPA groups. The overall continuation rates ranged from 70 to 86%. CONCLUSIONS: These results confirm that lower dose combinations of continuous combined HRT are usually sufficient to control symptoms or avoid breakthrough bleeding. However, if higher E(2)V dose is needed for symptom control, it should be combined with the higher dose of progestin (5 mg) to avoid bleeding disturbances. Flexible treatment regimens should be available for individualized HRT. PMID- 11731186 TI - Estrogen/progesterone replacement versus pravastatin and their sequential association in hypercholesterolemic postmenopausal women. AB - OBJECTIVES: The objectives of this study were to assess serum lipid changes in response to an oral estrogen combined with progesterone (Group A) as compared with pravastatin (Group B) and to evaluate the additive effects of the sequential addition of statin to hormonal replacement therapy (HRT) and of HRT to statin. METHODS: Thirty-seven of 63 hypercholesterolemic menopausal women initially submitted to a 4-month diet were randomised to oral conjugated estrogens (0.625 mg)/micronised progesterone (200 mg) or to pravastatin (40 mg). After 6 months, each group received both medications for another 6 months. RESULTS: Nineteen percent of women corrected their lipids below decision levels with diet alone. Low density lipoprotein-cholesterol (LDL-C) decreased by 8+/-5% with HRT and by 26+/-3% (P<0.001) with the statin. These single medications increased high density lipoprotein-cholesterol (HDL-C) by 13+/-5% (P<0.01) and 11+/-7%, respectively. Combined interventions produced cumulative LDL-C reductions of 40+/ 2 and 42+/-3% (P<0.001) and additive HDL-C augmentations of 16+/-4 and 23+/-5% (P<0.01) with proportional changes in apolipoprotein (Apo)B-100 and ApoA-1. These combined effects brought the atherogenic index (C/HDL-C) for Groups A and B, respectively, from a moderate (5.18+/-0.25 and 5.87+/-0.18) to a reduced (3.35+/ 0.20 and 3.52+/-0.19) risk category. Triglycerides (TG) which were increased by HRT and decreased by the statin returned to baseline during combined treatments. No changes in diet, physical activity or anthropomorphometric measurements explained the lipid modifications. CONCLUSIONS: In menopausal patients with elevated C not responding to diet, pravastatin was most effective to decrease LDL C, and oral estrogen-micronised progesterone most effective to increase HDL-C. Marked reduction of the atherogenic index is achieved by sequential combinations of medications resulting from beneficial cumulative effects on both C-LDL and C HDL. PMID- 11731188 TI - Proliferation profile of endometrial polyps in post-menopausal women. AB - OBJECTIVE: To determine whether or not the presence of c-erbB2 over-expression in endometrial polyps affects the percentage of cells positive for Ki-67 proliferation marker. METHODS: Thirty-five patients with endometrial polyps were submitted to polypectomy by hysteroscopy. Ki-67 and c-erbB2 over-expression were investigated in the polyps by immunohistochemistry. RESULTS: The presence of c erbB2 over-expression by immunohistochemistry was observed in 80% of endometrial polyps and was associated with higher proliferation rates as determined by the number of positive Ki-67 cell nuclei. In c-erbB2-negative polyps, the proliferation rates were low. CONCLUSION: Ki-67 and c-erbB2 over-expression are frequent in endometrial polyps in post-menopausal women. PMID- 11731189 TI - Errors in the estimation of the distance of fall and angles of impact blood drops. AB - This paper presents derived equations for the estimation of the variances in the estimated distance of fall and the estimated angle of impact for blood droplets. The derived equation for the variance of the estimated distance of fall predicts that as the blood drop size approaches that of a blood droplet falling at terminal velocity, the variance of the estimate grows without limit. The derived equation for the variance in the estimated angle of impact shows that as the angle of impact approaches 90 degrees, the variance grows without limit. The validity of the equation for the estimated variance of the angle of impact was tested for five angles of impact (15, 30, 45, 60 and 75 degrees); the equation correctly predicted the observed variance up to an angle of 60 degrees. PMID- 11731190 TI - Vacuum metal deposition: developing latent fingerprints on polyethylene substrates after the deposition of excess gold. AB - A previously identified difficulty with the development of latent fingerprints on low-density polyethylene (LDPE) by vacuum metal deposition (VMD) is that excess gold deposition prevents effective zinc deposition and so inhibits latent print development [1]. The investigation of a means to correct for excess gold deposition was the basis for this study. Exposure to zinc and the readmission of air into the vacuum chamber followed by VMD development results in the deactivation of the initial gold clusters due to the formation of zinc oxide and the adsorption of air and water molecules. As a result, the majority of the gold deposited during the second treatment creates new gold clusters rather than adding to the gold clusters previously formed. After excess gold deposition on LDPE, the deposition of 1.5 times the optimum gold amount will result in good quality normal development. The results also indicate that, on all surfaces, at least twice the initial gold count should be used for re-treatment when further development is required after air has been readmitted to the chamber. PMID- 11731191 TI - Acute intoxication with orphenadrine and clozapine. AB - This report describes a fatal intoxication with two different drugs: clozapine and orphenadrine. A 38-year-old man was found dead in the bedroom of his residence. Near the body were found various empty pharmaceutical boxes, employed in schizophrenic therapy, two of them containing clozapine and orphenadrine. High concentrations of clozapine and orphenadrine detected in blood, urine and gastric content were related to death. These compounds were identified and quantitated by liquid-liquid extraction followed by gas chromatographic/mass spectrometry (GC/MS) analysis. PMID- 11731192 TI - Determination of benzodiazepines in human hair by on-line high-performance liquid chromatography using a restricted access extraction column. AB - A method is described for the identification of five frequently prescribed benzodiazepines (BZD) (clonazepam, diazepam, flunitrazepam, midazolam and oxazepam) in human hair samples by reversed phase HPLC, following on-line simple enrichment and clean-up on a restricted access extraction column. 50mg of powdered hair were incubated (2h at 45 degrees C) after sonication (1h) in 1 ml of the following solution (methanol:ammonia, 97.5/2.5, v/v). The aliquot was centrifuged and the methanolic phase transferred to a conical tube and evaporated under a gentle stream of nitrogen. The residue was reconstituted by adding 100 microl of a mixture of phosphate buffer (20mM, pH=2.2) and acetonitrile (94/6, v/v). A total of 80 microl were injected into the system with the column switching technique. The pre-column or clean-up column was washed with phosphate buffer pH=7.2. The drugs retained on the pre-column were then eluted in the back flush mode and separated on a C(8) semi micro column, Lichrospher select B, 125 mm x 3 mm. The BZD were determined by a photodiode-array detector at 254 nm, using reference data (retention time and UV spectra) stored in a personal library. The method showed excellent linearity between 0.5 and 20 ng/mg of hair for clonazepam, flunitrazepam and midazolam and between 0.5 and 100 ng/mg of hair for diazepam and oxazepam. Finally, the present method has been applied to a number of forensic cases in our laboratory. PMID- 11731193 TI - Death due to baclofen and dipyrone ingestion. AB - A fatal suicidal intoxication with unusual drugs is reported. A 56-year-old man was found dead in his house; near by the corpse several empty drugs boxes were found. An autopsy was performed and the biological fluids were submitted to a full toxicological work-up. The analytical results supported the hypothesis of a death due to the acute baclofen (4-amino-3-(p-chlorophenyl)butyric acid) and dipyrone (sodium [N-(1,5-dimethyl-3-oxo-2-phenylpyrazolin-4-yl)-N-methylamino] methanesulfonate) intoxication. PMID- 11731194 TI - The STR-based genetic profile of the population from Corsica island (France). AB - Short tandem repeats (STR) at loci HumFES/FPS, HumVWA, HumCSF1PO, HumTH01, HumFXIIIA01, HumTPOX, HumCD4, D3S1358 are markers of choice for population genetics and validated systems for forensic use. In this report, we analysed their allele frequency distribution in a sample of native blood donors from the two departments of Corsica island (France). Deviations from the Hardy-Weinberg rule and heterozygosity values consistently suggested a spatial differentiation of allele and genotype frequencies across the island. Pairwise comparisons showed that Corsican gene pool presents a high level of heterogeneity between departments and substantially differs from that of neighbouring and historically related populations. The results suggest the use of local databases to calculate a priori statistics in human identity testing. PMID- 11731195 TI - Evaluation of the possibility of differentiation between various types of ammunition by means of GSR examination with SEM-EDX method. AB - Gunshot residues originating from six types of pistol ammunition were studied. Six persons who normally have no contact with weapons fired three times, each person from a different pistol. Samples of gunshot residues were collected from the shooters' hands using aluminium stubs with black carbon adhesive tabs and subjected to examinations of their morphology and elemental content by means of a scanning electron microscope with an energy dispersive X-ray spectrometer in an automatic manner. In a comparative study of the samples, only primer residues were taken into account. The frequency of occurrence of particles representing a certain chemical class against the total number of detected particles was expressed as a percentage. Mutual relations between the frequencies of occurrence of the residues of particular chemical content were found to be different for most of the studied ammunition types using a non-statistical approach--and by means of non-parametric statistical methods (R-Spearman and tau-Kendall correlation coefficients), it was possible to differentiate one ammunition type from each of the other ammunition types examined. The performed examinations have revealed some differences in the frequencies of occurrence of certain chemical classes of primer residues, observed for various types of ammunition. These differences could constitute a valuable contribution to group identifications of ammunition made on the basis of physical and chemical examinations of gunshot residues. PMID- 11731196 TI - A textile fibre survey as an aid to the interpretation of fibre evidence in the Sydney region. AB - Frequency figures of the fibre population on textile cinema seats were measured in Sydney, Australia, in winter. Sixteen seats were analysed from a very popular cinema complex, with 3025 fibres classified according to colour, generic class and fluorescence properties (100 grey-black cotton fibres only). The recovered fibres were mostly natural fibres (84%) with cotton the most common generic type (70%). On the contrary, man made fibres were relatively rare (15%) with rayon constituting the majority of these (51%). The most common colour/generic class combinations were grey-black cotton (33%) and blue cotton (30%) accounting for 63% of the total population. All other frequencies were below 5%, most below 1% using only the two properties of colour and generic class. Fluorescence properties were found to be very discriminating as far as grey-black cotton fibres were concerned. These features are considered and discussed and in particular, to emphasise the significance of fibres as evidence of contact. PMID- 11731197 TI - Body models in forensic ballistics: reconstruction of a gunshot injury to the chest by bullet fragmentation after shooting through a finger. AB - Forensic science uses substitutes to reconstruct injury patterns in order to answer questions regarding the dynamic formation of unusual injuries. Using a case study, an experimental simulation of a finger was designed, for the first time with a combination of hard wood and glycerin soap. With this model as an intermediate target simulation, it was possible not only to demonstrate the "bullet-body (finger) interaction", but also to recreate the wound pattern found in the victim. This case demonstrates that by using ballistic models and body part substitutes, gunshot cases can be reproduced simply and economically, without coming into conflict with ethical guidelines. PMID- 11731198 TI - Prevalence of potential risk factors in victims of electrocution. AB - OBJECTIVE: To identify risk factors of fatal arrhythmia following electrical shock, by comparing the prevalence of transthoracic current, tetany, decreased skin resistance because of wet extremities, skin burns and heart disease in victims of electrocution with instant as opposed to delayed death. DESIGN: Retrospective case-control study, based on the charts from the coroner's office. RESULTS: A total of 124 deaths from electrocution occurred between 1987 and 1992. One victim presumably died from delayed arrhythmia and was excluded from the study. Twenty victims had decreased skin resistance because of wet extremities and five had tetany. Autopsy revealed coronary heart disease in 21 cases, and burns in 109; 10 did not have any skin lesion. There was no difference in risk factors between those who died instantly presumably from arrhythmia (n=114) and those who died later from other causes (n=9). CONCLUSION: No differences in risk factors were found between victims who died immediately from arrhythmia following electrical shock and those died later from other causes. PMID- 11731199 TI - Fatal poisoning in drug addicts in the Nordic countries. AB - The study includes medicolegally examined fatal poisonings among drug addicts in 1997 in the five Nordic countries: Denmark, Finland, Iceland, Norway and Sweden, and the results are compared to a similar investigation from 1991. A common definition of "drug addict" was applied by the participating countries. The highest death rate by poisoning in drug addicts was observed in Denmark, where it was 6.54 per 10(5)inhabitants, followed by Norway with 6.35, Sweden with 2.21, Finland with 1.63 and Iceland with 1.20 per 10(5)inhabitants. All countries showed a higher death rate in 1997 than in 1991. For all countries the distribution of deaths according to geographical regions showed a decreasing number of drug deaths in the metropolitan area and an increasing number in other cities. Heroin/morphine dominated as the cause of death and was responsible for about 90% of the cases in Norway. In Sweden and Denmark, however, heroin/morphine caused only about 70% of the fatal poisonings. About 30% of the fatal poisonings in Denmark and Sweden were caused by other group I drugs, in Denmark mainly methadone and in Sweden mainly propoxyphene. Apart from two cases in Sweden methadone deaths were not seen in the other Nordic countries. In Finland heroin/morphine deaths have increased from about 10% in 1991 to about 40% in 1997. Forty-four percent of the fatal poisonings in Finland were caused by other group I drugs, mainly codeine and propoxyphene. The two fatal poisonings in Iceland were caused by carbon monoxide. Only few deaths in this investigation were caused by amphetamine and cocaine. A widespread use of alcohol, cannabis and benzodiazepines, especially diazepam, was seen in all the countries. PMID- 11731200 TI - Italian population data for D1S1656, D3S1358, D8S1132, D10S2325, VWA, FES/FPS, and F13A01. AB - Allele frequencies for seven STRs loci were obtained from a sample of 215 unrelated healthy Italian individuals. PMID- 11731201 TI - STR data for the AmpFlSTR profiler plus loci from Macau (China). AB - Allele frequencies for the nine STRs included in the AmpFlSTR profiler plus kit (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820) were estimated from a sample of 87 unrelated individuals living in the region of Macau, a territory located in the southeastern coast of China. PMID- 11731202 TI - STR data (CD4, CSF1PO, F13A01, FES/FPS, MBPB, TH01, TPOX) from North Portugal. AB - Allele frequencies for seven STRs (CD4, CSF1PO, F13A01, FES/FPS, MBPB, TH01, TPOX) were estimated from samples (sized between 300 and 940) of unrelated individuals born in North Portugal. PMID- 11731203 TI - 7-Locus Y chromosome haplotype profiling in a northern Spain population. AB - Allele and haplotype frequencies for 7 Y-specific STR loci (DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393) had been determined in a sample of 107 unrelated males living in Cantabria, a region in northern Spain, by means of two multiplex PCRs. PMID- 11731204 TI - Congenital skeletal disorders. PMID- 11731205 TI - The diagnosis of skeletal dysplasias: a multidisciplinary approach. AB - Skeletal dysplasias are heritable connective tissue disorders affecting skeletal morphogenesis and development. They represent a heterogeneous group of genetic disorders with more than 200 different entities being delineated to date. Because of this diversity, the diagnosis of a skeletal dysplasia is usually based on a combination of clinical, radiographic, morphologic, and, in some instances, biochemical and molecular studies. Tremendous advances have been made in the elucidation of the genetic defect of several of these conditions over the past 10 years. This progress has provided us with more insights into the genes controlling normal skeletal development. It also has opened new diagnostic perspectives. For several disorders, identification of the causal gene allows us now to confirm with a molecular test the diagnosis postulated on the basis of clinical, radiographic and/or morphologic studies. It also enables us to establish the diagnosis early in pregnancy. An accurate diagnosis is not only important for proper management of the affected individual but also the cornerstone for adequate genetic counseling. PMID- 11731206 TI - Congenital skeletal abnormalities: an introduction to the radiological semiology. AB - Despite the recent advances in the molecular diagnosis of congenital abnormalities, the initial identification and the decision to refer a patient for further molecular analysis and expensive genetic tests still relies frequently on clinical and radiological criteria. The radiological identification of syndromes, dwarfs and dysplasias is a difficult task, because there are so many findings to consider and so many syndromes to remember that the problem is overwhelming. There is a definite need for an easy and systematic analysis system, in order to try to categorize a skeletal dysplasia in a certain group. In this brief review, we suggest an approach to the evaluation of skeletal syndromes, based on the analysis of cardinal criteria, from which the most useful information is derived, and additional criteria, making further differentiation possible. Generally, cardinal information is derived from analysis of the long bones, hands, pelvis and the spine, whereas the analysis of other skeletal elements, like the skull, feet, and other flat bones is of additional value. PMID- 11731207 TI - Hereditary neuromuscular diseases. AB - This article presents the actual classification of neuromuscular diseases based on present expansion of our knowledge and understanding due to genetic developments. It summarizes the genetic and clinical presentations of each disorder together with CT findings, which we studied in a large group of patients with neuromuscular diseases. The muscular dystrophies as the largest and most common group of hereditary muscle diseases will be highlighted by giving detailed information about the role of CT and MRI in the differential diagnosis. The radiological features of neuromuscular diseases are atrophy, hypertrophy, pseudohypertrophy and fatty infiltration of muscles on a selective basis. Although the patterns and distribution of involvement are characteristic in some of the diseases, the definition of the type of disease based on CT scan only is not always possible. PMID- 11731208 TI - Molecular and radiological diagnosis of sclerosing bone dysplasias. AB - Bone mineral density (BMD) is a quantitative trait for which the heritability of the variance is estimated to be up to 80%, based on epidemiological and twin studies. Further illustration of the involvement of genetic factors in bone homeostasis, is the existence of an extended group of genetic conditions associated with an abnormal bone density. The group of conditions with increased bone density has long been poorly studied and understood at the molecular genetic level but recently, thanks to recent developments in molecular genetics and genomics, for some of them major breakthroughs have been made. These findings will make the molecular analysis of such patients an additional tool in diagnostics and in genetic counseling. However, the initial identification of affected patients is still largely dependent upon recognition of clinical and radiological stigmata of the disease. Therefore, in this overview of sclerosing bone dysplasias, the classical clinical and radiological signs of this group of disorders will be discussed along with the new molecular insights. PMID- 11731209 TI - Hereditary multiple exostoses: from genetics to clinical syndrome and complications. AB - OBJECTIVE: To give an overview of genetic, clinical and radiological aspects in two families over four generations with known hereditary multiple exostoses (HME). METHODS AND MATERIAL: After linkage analysis in both families to localize the defective gene, mutation analysis was performed in these genes to identify the underlying mutation. In the 31 affected individuals, location, number and morphology and evolution of exostosis, evolution of remodeling defects at the metaphysis, and the extent of possible complications were evaluated on clinical and imaging (plain radiography, computed tomography (CT), and magnetic resonance imaging (MRI)) data over a lifetime period. RESULTS AND CONCLUSIONS: Both families demonstrate the gene defect in the same EXT-2 gene locus on chromosome 11p. Exostoses are preferentially located in the lower extremity (hip, knee and lower leg), humerus, and forearm. Any other bone may be involved, except for the calvaria of the skull and the mandible. Exostoses are rather sessile than pedunculated. Exostosis is rarely present at birth but develops gradually and may persist to grow slowly after closure of the growth plates. Preferential expression of the remodeling defect was seen in the hip, distal femur (trumpet shaped metaphysis) and forearm (shortening of the ulna with secondary bowing of the radius and development of a pseudo-Madelung deformity). These radiological manifestations start at the age of 4-5 years and become more obvious as the enchondral bone formation progresses with age. Reported complications in these families consist of local entrapment phenomenons (vessel, tendon, nerve), frictional bursitis, and sarcomatous transformation. MRI was able to suggest these complications and is the imaging technique of choice in the evaluation of symptomatic exostoses. PMID- 11731210 TI - Introduction of skull bone fragments into the brain during external ventricular drain placement. AB - OBJECTIVE: To report an observation of small high attenuation foci in the tract of external ventricular drain on computed tomographic (CT) scan after removal of the drain and propose an explanation for this observation. METHODS: Six patients with small high attenuation foci in the ventricular drain tract seen on CT after removal of the drain were retrospectively identified. The CT studies before, during, and after ventricular drainage were reviewed for the size, shape, location, and time of appearance of these foci. RESULTS: In all patients, the high attenuation foci in the drain's tract were identified on CT done within 24 h from removal of the drain. In one patient, the high attenuation focus was seen adjacent to the drain on the CT obtained while the drain was in place. The high attenuation foci maintained constant size and shape on serial follow-up CT, and had variable location along the drain's tract. None of the patients developed symptoms related to these foci. CONCLUSION: Small bone pieces from the calvarium after drilling for external ventricular drain can inadvertently be pushed into the brain parenchyma during drain insertion. Early identification of these pieces on CT after removal of the drain and their constant size and shape are the clues for their nature. Temporal evolution of CT findings does not support the possibility of calcification in a small infarction or hematoma in the tract of the drain. PMID- 11731211 TI - MR imaging features of bizarre parosteal osteochondromatous proliferation of bone (Nora's lesion). AB - The purpose of this study was to review the imaging findings of three patients with bizarre parosteal osteochrondromatous proliferation of bone (BPOP). The plain radiographs and MRI images of three patients with BPOP were obtained and retrospectively reviewed. In two cases, BPOP involved the feet. In one case BPOP involved the hand. In all three cases, plain radiographs showed a well-defined calcium containing mass adjacent to the cortical surface of the adjacent bone. The underlying bone appeared normal in all cases. On MRI, the lesion was of low signal intensity on T1 weighted sequences in all cases. On FSE T2 weighted and STIR sequences, the lesion was of high signal in all cases. The cortex, medullary cavity and adjacent soft tissues appeared normal in all cases. While BPOP is rare and often confused with a variety of both benign and malignant lesions, there are specific radiological findings that may help to distinguish BPOP from many of its mimickers. PMID- 11731212 TI - Tuberculous arthritis mimic arthritis of the Sjogren's syndrome: findings from sonography, computed tomography and magnetic resonance images. AB - A patient with a history of Sjogren's syndrome developed chronic arthritis of left ankle. It was diagnosed as arthritis of the Sjogren's syndrome initially. However, joint pain persisted despite corticosteroid therapy. Sonography disclosed a multiloculated cystic lesion with peripheral hyperechoic enhancement around left ankle and extended to Achilles tendon and subcutaneous region. Computed tomography (CT) confirmed the findings. Magnetic resonance imaging (MRI) revealed increased signal intensity of the lesion after gadonillium enhancement on T1-weighted images. These abnormalities showed inhomogenous high signal intensities on T2-weighted images. Tuberculous arthritis was diagnosed by positive synovial tuberculous culture. Sonography is a valuable tool that offers significant advantages for the initial evaluation of arthritis of the Sjogren's syndrome and help early suspicious of tuberculous arthritis, because of its cost effectiveness, superior differentiation between the cyst and solid lesions, convenience for guiding biopsy and drainage. PMID- 11731213 TI - Sacro-iliac joint arthrography in low back pain: feasibility of MRI guidance. AB - OBJECTIVE: Evaluation of the feasibility of MR-guidance in sacro-iliac joint arthrography in patients whose low back pain is suspected to arise from the sacro iliac joint (SIJ). METHODS AND PATIENTS: Twenty patients with low back pain underwent MR-guided sacro-iliac joint arthrography. Needles made from titanium (size 20 G, MDTech, USA) were used. For image guidance a 0.23T open-configuration C arm magnet (Proview, Marconi Medical Systems, USA) with special interventional hardware and software package (I-Path 200, Marconi Medical Systems) containing an MR compatible in-room console, large-screen (36") display, optical navigator and accompanying software with dedicated sequences was used. No other image guidance modalities were used. RESULTS: MR-guided SIJ arthrography was successfully performed in all twenty patients without complications. The optimal imaging sequence both for preoperative and intraoperative images was 3D-Gradient Echo sequence. The dispersal of the injected saline and anaesthetic could be determined inside the joint in all cases with heavily T2-weighted fast spin echo sequence. In the present study, 60% of the patients had significant reduction of pain after sacro-iliac joint arthrography and sacro-iliac joint was considered to be the source of patients low back pain in these patients. CONCLUSION: The present study shows that MR guidance with open configuration low field scanner is an accurate guiding method for sacro-iliac joint arthrography. PMID- 11731214 TI - The oblique needle technique in lumbar facet joint injection. AB - OBJECTIVE: To introduce and evaluate an alternative technique for accessing the lumbar facet joints. This technique uses an oblique approach to enter the facet joint without the need to rotate the patient towards the beam. METHODS AND MATERIAL: The proposed technique was employed in 23 consecutive patients (50 joints) with diagnosed or suspected facet joint syndrome. We recorded the success rate, the time spent to complete the procedure, and any complications arisen. RESULTS: The overall success rate was 84% and the average time spent for the procedure was 4.8 min. No complications were encountered. CONCLUSIONS: Due to its inherent simplicity the proposed technique is easily tolerated by the patient, has a high success rate and can be performed quickly without complications. PMID- 11731215 TI - Sepsis from dropped clips at laparoscopic cholecystectomy. AB - We report seven patients in whom five dropped surgical clips and two gallstones were visualized in the peritoneal cavity, on radiological studies. In two, subphrenic abscesses and empyemas developed as a result of dropped clips into the peritoneal cavity during or following laparoscopic cholecystectomy. In one of these two, a clip was removed surgically from the site of an abscess. In two other patients dropped gallstones, and in three, dropped clips led to no complications. These were seen incidentally on studies done for other indications. Abdominal abscess secondary to dropped gallstones is a well recognized complication of laparoscopic cholecystectomy (LC). We conclude that even though dropped surgical clips usually do not cause problems, they should be considered as a risk additional to other well-known causes of post-LC abdominal sepsis. PMID- 11731220 TI - SLC19A3 encodes a second thiamine transporter ThTr2. AB - Recently, a new family of facilitative carriers has been cloned consisting of the reduced folate (SLC19A1) and the thiamine (SLC19A2) transporters. Despite a high level of sequence identity and similarity there is essentially no functional overlap between these carriers. The former transports folates and the latter thiamine. In this paper we describe the function of SLC19A3, another member of this transporter family most recently cloned, after transient transfection of the cDNA into HeLa cells. Uptake of [3H]thiamine, but not of methotrexate nor folic acid, was enhanced in SLC19A3 transfectants relative to vector control. Similarly, in the transfectants thiamine transport increased with an increase in pH with peak activity at pH approximately 7.5. While [3H]thiamine uptake was markedly inhibited by nonlabeled thiamine it was not inhibited by several organic cations in 100-fold excess. Hence this carrier has a high degree of specificity for vitamin B1. The data indicate that SLC19A3 has the characteristics of SLC19A2 (ThTr1) and represents a second thiamine transporter (ThTr2) in this family of facilitative carriers. PMID- 11731221 TI - Islet amyloid and type 2 diabetes: from molecular misfolding to islet pathophysiology. AB - Islet amyloid polypeptide (IAPP, amylin) is secreted from pancreatic islet beta cells and converted to amyloid deposits in type 2 diabetes. Conversion from soluble monomer, IAPP 1-37, to beta-sheet fibrils involves changes in the molecular conformation, cellular biochemistry and diabetes-related factors. In addition to the recognised amyloidogenic region, human IAPP (hIAPP) 20-29, the peptides human or rat IAPP 30-37 and 8-20, assume beta-conformation and form fibrils. These three amyloidogenic regions of hIAPP can be modelled as a folding intermediate with an intramolecular beta-sheet. A hypothesis is proposed for co secretion of proIAPP with proinsulin in diabetes and formation of a 'nidus' adjacent to islet capillaries for subsequent accumulation of secreted IAPP to form the deposit. Although intracellular fibrils have been identified in experimental systems, extracellular deposition predominates in animal models and man. Extensive fibril accumulations replace islet cells. The molecular species of IAPP that is cytotoxic remains controversial. However, since fibrils form invaginations in cell membranes, small non-toxic IAPP fibrillar or amorphous accumulations could affect beta-cell stimulus-secretion coupling. The level of production of hIAPP is important but not a primary factor in islet amyloidosis; there is little evidence for inappropriate IAPP hypersecretion in type 2 diabetes and amyloid formation is generated in transgenic mice overexpressing the gene for human IAPP only against a background of obesity. Animal models of islet amyloidosis suggest that diabetes is induced by the deposits whereas in man, fibril formation appears to result from diabetes-associated islet dysfunction. Islet secretory failure results from progressive amyloidosis which provides a target for new therapeutic interventions. PMID- 11731222 TI - Risk of iron overload is decreased in beating heart coronary artery surgery compared to conventional bypass. AB - Conventional cardiopulmonary bypass surgery (CCPB) increases the iron loading of plasma transferrin often to a state of plasma iron overload, with the presence of low molecular mass iron. Such iron is a potential risk factor for oxidative stress and microbial virulence. Here we assess 'off-pump' coronary artery surgery on the beating heart for changes in plasma iron chemistry. Seventeen patients undergoing cardiac surgery using the 'Octopus' myocardial wall stabilisation device were monitored at five time points for changes in plasma iron chemistry. This group was further divided into those (n=9) who had one- or two- (n=8) vessel grafts, and compared with eight patients undergoing conventional coronary artery surgery. Patients undergoing beating heart surgery had significantly lower levels of total plasma non-haem iron, and a decreased percentage saturation of their transferrin at all time points compared to conventional bypass patients. Plasma iron overload occurred in only one patient undergoing CCPB. Beating heart surgery appears to decrease red blood cell haemolysis, and tissue damage during the operative procedures and thereby significantly decreases the risk of plasma iron overload associated with conventional bypass. PMID- 11731223 TI - The basal kinetic parameters of glycogen synthase in human myotube cultures are not affected by chronic high insulin exposure. AB - There is no consensus regarding the results from in vivo and in vitro studies on the impact of chronic high insulin and/or high glucose exposure on acute insulin stimulation of glycogen synthase (GS) kinetic parameters in human skeletal muscle. The aim of this study was to evaluate the kinetic parameters of glycogen synthase activity in human myotube cultures at conditions of chronic high insulin combined or not with high glucose exposure, before and after a subsequent acute insulin stimulation. Acute insulin stimulation significantly increased the fractional activity (FV(0.1)) of GS, increased the sensitivity of GS to the allosteric activator glucose 6-phosphate (A(0.5)) and increased the sensitivity of GS to its substrate UDPG (K(m(0.1))) when myotubes were precultured at low insulin with/without high glucose conditions. However, this effect of acute insulin stimulation was abolished in myotubes precultured at high insulin with or without high glucose. Furthermore, we found significant correlations between the fractional velocities FV(0.1) of GS and K(m(0.1)) (rho=-0.72, P<0.0001), between FV(0.1) and A(0.5) (rho=-0.82, P<0.0001) and between K(m(0.1)) and A(0.5) values (rho=0.71, P<0.0001). Our results show that chronic exposure of human myotubes to high insulin with or without high glucose did not affect the basal kinetic parameters but abolished the reactivity of GS to acute insulin stimulation. We suggest that insulin induced insulin resistance of GS is caused by a failure of acute insulin stimulation to decrease A(0.5) and K(m(0.1)) in human skeletal muscle. PMID- 11731224 TI - Butylamino-demethoxy-hypocrellins and photodynamic therapy decreases human cancer in vitro and in vivo. AB - 2-Butylamino-2-demethoxy-hypocrellin A (BAHA) and B (BAHB) are new photosensitizers synthesized by a mild reaction of hypocrellins and butylamine. In BAHA and BAHB, the peri-hydroxylated perylenequinone structure of the parent hypocrellins is preserved and the red absorption is enhanced distinctly. Electron paramagnetic resonance spin trapping measurements and 9,10-diphenylanthracene bleaching studies were used to investigate the photodynamic action of BAHA and BAHB in the presence of oxygen. Singlet oxygen (1O2) and superoxide anion radical (O2(*-)) produced by illuminating BAHA and BAHB in aerobic solution have been observed. Compared with hypocrellin A and B, BAHA and BAHB primarily remained able to generate 1O2 and enhanced distinctly the O2(*-)-generating abilities. The photodynamic action of BAHA and BAHB in the therapy of cancer was investigated in vitro and in vivo. Both in vitro and in vivo results revealed a significant decrease in cancer cell growth. Laser or dye alone had no effect, indicating that intratumor BAHA and laser therapy may prove useful in unresectable cancer. PMID- 11731225 TI - The effect of four mutations on the expression of iduronate-2-sulfatase in mucopolysaccharidosis type II. AB - Mucopolysaccharidosis type II (Hunter syndrome; OMIM 309900) is a rare X-linked recessive lysosomal storage disorder caused by the deficiency of the enzyme iduronate-2-sulfatase (IDS; EC 3.1.6.13). Different alterations at the IDS locus, mostly missense mutations, have been demonstrated, by expression study, as deleterious, causing significant consequences on the enzyme function or stability. In the present study we report on the results of the transient expression of the novel K347T, 533delTT, N265I and the already described 473delTCC (previously named DeltaS117) mutations in the COS 7 cells proving their functional consequence on IDS activity. This type of information is potentially useful for genotype-phenotype correlation, prognosis and possible therapeutic intervention. PMID- 11731226 TI - Variation in the promoter of the human hormone sensitive lipase gene shows gender specific effects on insulin and lipid levels: results from the Ely study. AB - We previously identified a hormone sensitive lipase (HSL) promoter variant, 60C>G, which in vitro exhibits 40% reduced promoter activity. In this study we examined the effect of the -60C>G on glycemic and lipid measures in the population based Ely study of metabolic function and insulin resistance in 218 middle-aged men and 276 middle-aged women. Adipose tissue HSL is the rate limiting step in triglyceride lipolysis, generating free fatty acids for energy utilization. HSL is also expressed in pancreatic beta-cells where its activity therefore may affect insulin secretion. In the women, carriers of the HSL -60G allele had significantly lower fasting insulin levels (P=0.0005) and a lower total area under the curve for insulin during the oral glucose tolerance test (P=0.005). There was no demonstrable association in men with these measures of insulin sensitivity but carriers of the -60G allele had significantly lower fasting non-esterified fatty acid (NEFA) levels (P=0.025) and higher low density lipoprotein cholesterol levels (P=0.02) than men who were non-carriers. This study provides additional evidence for a role for HSL in the development of insulin resistance, from which carriers of the -60G allele, associated here with markers of insulin sensitivity in women, and with lower NEFA levels in men, might be protected. PMID- 11731227 TI - Multiple fibroblast growth factors support growth of the ureteric bud but have different effects on branching morphogenesis. AB - Together with glial-derived neurotrophic factor (GDNF), soluble factors present in a metanephric mesenchyme (MM) cell conditioned medium (BSN-CM) are necessary to induce branching morphogenesis of the isolated ureteric bud (UB) in vitro (Proc. Natl. Acad. Sci. USA 96 (1999) 7330). Several lines of evidence are presented here in support of a modulating role for fibroblast growth factors (FGFs) in this process. RT-PCR revealed the expression of two FGF receptors, FGFR1(IIIc) and FGFR2(IIIb), in isolated embryonic day 13 rat UBs, which by indirect immunofluorescence displayed a uniform distribution. Rat kidney organ culture experiments in the presence of a soluble FGFR2(IIIb) chimera or a neutralizing antibody to FGF7 suggested an important contribution of FGFs other than FGF7 to the branching program. Several FGFs, including FGF1, FGF2, FGF7 and FGF10, in combination with GDNF and BSN-CM were found to affect growth and branching of the isolated UB, albeit with very different effects. FGF1 and FGF7 were at extreme ends of the spectrum, with FGF10 (more FGF1-like) and FGF2 (more FGF7-like) falling in between. FGF1 induced the formation of elongated UB branching stalks with distinct proliferative ampullary tips, whereas FGF7 induced amorphous buds displaying nonselective proliferation with little distinction between stalks and ampullae. Electron microscopic examination demonstrated that FGF1 treatment induced cytoskeletal organization, intercellular junctions and lumens along the stalk portion of the developing tubules, while the ampullary regions contained 'less differentiated' cells with an abundant secretory apparatus. In contrast, FGF7-induced UBs displayed this 'less differentiated' morphology regardless of position on the structure and were virtually indistinguishable from FGF1-induced ampullae. Consistent with this, GeneChip array analysis (employing a novel nanogram-scale assay consisting of two rounds of amplification and in vitro transcription for analyzing small quantities of RNA) revealed that FGF7-induced UBs expressed more markers of cell proliferation than FGF1, which caused the UB to express cytoskeletal proteins, extracellular matrix proteins, and at least one integrin, some of which may be important in UB branch elongation. Thus, while the various FGFs examined all support UB growth, FGF1 and FGF10 appear to be more important for branching and branch elongation, and may thus play a role in determination of nephron number and patterning in the developing kidney. These in vitro data may help to explain results from knockout and transgenic studies and suggest how different FGFs may, together with GDNF and other factor(s) secreted by MM cells, regulate branching morphogenesis of the UB by their relative effects on its growth, branching and branch elongation and differentiation, thereby affecting patterning in the developing kidney. PMID- 11731228 TI - The Drosophila RGS protein Loco is required for dorsal/ventral axis formation of the egg and embryo, and nurse cell dumping. AB - The loco gene encodes members of a family of RGS proteins responsible for the negative regulation of G-protein signalling. At least two transcripts of loco are expressed in oogenesis, loco-c2 is observed in the anterior-dorsal follicle cells and is downstream of the epidermal growth factor receptor signalling pathway, initiated in the oocyte. loco-c3 is a new transcript of loco, which is expressed in the nurse cells from stage 6 onwards. Analysis of newly generated mutants and antisense technology enabled us to establish that disrupting loco in follicle cells results in ventralized eggs, while disrupting loco in nurse cells results in short eggs, due to defective dumping of the nurse cell cytoplasm into the oocyte. PMID- 11731229 TI - FAM deubiquitylating enzyme is essential for preimplantation mouse embryo development. AB - FAM is a developmentally regulated substrate-specific deubiquitylating enzyme. It binds the cell adhesion and signalling molecules beta-catenin and AF-6 in vitro, and stabilises both in mammalian cell culture. To determine if FAM is required at the earliest stages of mouse development we examined its expression and function in preimplantation mouse embryos. FAM is expressed at all stages of preimplantation development from ovulation to implantation. Exposure of two-cell embryos to FAM-specific antisense, but not sense, oligodeoxynucleotides resulted in depletion of the FAM protein and failure of the embryos to develop to blastocysts. Loss of FAM had two physiological effects, namely, a decrease in cleavage rate and an inhibition of cell adhesive events. Depletion of FAM protein was mirrored by a loss of beta-catenin such that very little of either protein remained following 72h culture. The residual beta-catenin was localised to sites of cell-cell contact suggesting that the cytoplasmic pool of beta-catenin is stabilised by FAM. Although AF-6 levels initially decreased they returned to normal. However, the nascent protein was mislocalised at the apical surface of blastomeres. Therefore FAM is required for preimplantation mouse embryo development and regulates beta-catenin and AF-6 in vivo. PMID- 11731230 TI - Interaction of gap genes in the Drosophila head: tailless regulates expression of empty spiracles in early embryonic patterning and brain development. AB - Unlike gap genes in the trunk region of Drosophila embryos, gap genes in the head were presumed not to regulate each other's transcription. Here, we show that in tailless (tll) loss-of-function mutants the empty spiracles (ems) expression domain in the head expands, whereas it retracts in tll gain-of-function embryos. We have identified a 304bp element in the ems-enhancer which is sufficient to drive expression in the head and brain and which contains two TLL and two BCD binding sites. Transgenic reporter gene lines containing mutations of the TLL binding sites demonstrate that tll directly inhibits the expression of ems in the early embryonic head and the protocerebral brain anlage. These results are the first demonstration of direct transcriptional regulation between gap genes in the head. PMID- 11731231 TI - Dynamic expression and nuclear accumulation of beta-catenin during the development of hair follicle-derived structures. AB - Beta-catenin has a dual role in the cell. At the membrane, it connects E-cadherin to the actin cytoskeleton, while in the nucleus, it controls gene expression in concert with Tcf-like transcription factors. Nuclear translocation of beta catenin is induced by the Wnt signal transduction pathway. Control of this process is essential since elevated beta-catenin levels interfere with differentiation and development, and can initiate cancer in many tissues. An important role for beta-catenin during hair follicle related development and tumorigenesis has recently been established, though little is known of its endogenous expression during the development of these structures. Here, we have investigated the expression of beta-catenin in relation to markers for proliferation, differentiation and Wnt signaling during the development of three hair follicle related structures, i.e. whiskers, normal body hair and the preputial gland, and a hair follicle-derived tumor, the epidermal cyst. We observed nuclear accumulation of beta-catenin, the hallmark of Wnt signaling, in the upper matrix, the dermal papilla, the developing ringwulst of the whisker and in the tumor, though it was never in association with proliferation or terminal differentiation. Co-localization of nuclear beta-catenin with Tcf-3/4 was found only in the dermal papilla and the developing ringwulst of the whisker, but not in the upper matrix or in the tumor. These results further elucidate the role of the Wnt signal transduction pathway during hair follicle related development and tumorigenesis and illustrate the dynamic role of beta-catenin in signal transduction and cell-adhesion. PMID- 11731232 TI - Expression of HNF4alpha isoforms in mouse liver development is regulated by sequential promoter usage and constitutive 3' end splicing. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) is essential for the establishment and maintenance of liver-specific gene expression. The HNF4alpha gene codes for several isoforms whose developmental and physiological relevance has not yet been explored. HNF4alpha1 and HNF4alpha7 originate from different promoters, while alternative splicing in 3' leads to HNF4alpha2 and HNF4alpha8, respectively. HNF4alpha7/alpha8 were abundantly expressed in embryonic liver and fetal-like hepatoma cells. HNF4alpha1/alpha2 transcripts were up-regulated at birth and represented the only isoforms in adult-like hepatoma cells. In line with its expression profile, HNF4alpha7 activated more avidly than HNF4alpha1 reporter plasmids for genes that are expressed early. The expression patterns of both isoforms together with the differences observed in their transcriptional activities provide elements accounting for fine-tuning of the activity of HNF4alpha. The sequential expression of HNF4alpha7/alpha8 and HNF4alpha1/alpha2 during mouse liver development is the only modification in liver-enriched transcription factors thus far recorded, which parallels the transition from the fetal to the adult hepatic phenotype. PMID- 11731233 TI - SNT-1/FRS2alpha physically interacts with Laloo and mediates mesoderm induction by fibroblast growth factor. AB - Members of the fibroblast growth factor (FGF) ligand family play a critical role in mesoderm formation in the frog Xenopus laevis. While many components of the signaling cascade triggered by FGF receptor activation have been identified, links between these intracellular factors and the receptor itself have been difficult to establish. We report here the characterization of Xenopus SNT-1 (FRS2alpha), a scaffolding protein previously identified as a mediator of FGF activity in other biological contexts. SNT-1 is widely expressed during early Xenopus development, consistent with a role for this protein in mesoderm formation. Ectopic SNT-1 induces mesoderm in Xenopus ectodermal explants, synergizes with low levels of FGF, and is blocked by inhibition of Ras activity, suggesting that SNT-1 functions to transmit signals from the FGF receptor during mesoderm formation. Furthermore, dominant-inhibitory SNT-1 mutants inhibit mesoderm induction by FGF, suggesting that SNT-1 is required for this process. Expression of dominant-negative SNT-1 in intact embryos blocks mesoderm formation and dramatically disrupts trunk and tail development, indicating a requirement for SNT-1, or a related factor inhibited by the mutant construct, during axis formation in vivo. Finally, we demonstrate that SNT-1 physically associates with the Src-like kinase Laloo, and that SNT-1 activity is required for mesoderm induction by Laloo, suggesting that SNT-1 and Laloo function as components of a signaling complex during mesoderm formation in the vertebrate. PMID- 11731234 TI - Skin-derived nerve growth factor blocks programmed cell death in the trigeminal ganglia but does not enhance neuron proliferation. AB - Development of the cutaneous sensory nervous system is dependent on the production of neurotrophic factors, such as nerve growth factor (NGF), by the skin. Limited synthesis of NGF in developing skin is thought to underlie programmed cell death and cause a 50% neuronal loss. This loss does not occur in transgenic mice that overexpress NGF in the skin, which have double the number of neurons (J. Neurosci. 14 (1994) 1422). To determine whether increased NGF blocks neuronal death and/or increases neuronal precursor replication, we analyzed the trigeminal ganglia at embryonic days E12.5, E14.5 and E16.5 using transferase mediated dUTP nick-end labeling (TUNEL) and bromodeoxyuridine labeling. Results show that excess target-derived NGF causes a major decrease in the percent of TUNEL-labeled neurons without affecting the percent of replicating neurons. Analysis of RNA and protein expression suggests this block in cell death is mediated via the anti-apoptotic protein bcl-2. PMID- 11731235 TI - The transcription factor Pax6 is required for development of the diencephalic dorsal midline secretory radial glia that form the subcommissural organ. AB - During brain development, Pax6 is expressed in specific regions of the diencephalon including secretory cells of the subcommissural organ (SCO), a circumventricular organ at the forebrain-midbrain boundary that originates from the pretectal dorsal midline neuroepithelial cells beneath the posterior commissure (PC). Homozygous small eye (Sey/Sey) mice lack functional Pax6 protein and fail to develop the SCO, a normal PC and the pineal gland. Small eye heterozygotes (Sey/+) show defective development of the SCO's basal processes which normally penetrate the PC, indicating that normal development of the gland requires normal Pax6 gene-dosage. A correlation between the defects of SCO formation and altered R- and OB-cadherin expression patterns in the SCO is observed in mutants suggesting a role for cadherins in SCO development. PMID- 11731236 TI - Early patterning of the prospective midbrain-hindbrain boundary by the HES related gene XHR1 in Xenopus embryos. AB - The molecular mechanisms that govern early patterning of anterior neuroectoderm (ANE) for the prospective brain region in vertebrates are largely unknown. Screening a cDNA library of Xenopus ANE led to the isolation of a Hairy and Enhancer of split- (HES)-related transcriptional repressor gene, Xenopus HES related 1 (XHR1). XHR1 is specifically expressed in the midbrain-hindbrain boundary (MHB) region at the tailbud stage. The localized expression of XHR1 was detected as early as the early gastrula stage in the presumptive MHB region, an area just anterior to the involuting dorsal mesoderm that is demarcated by the expression of the gene Xbra. Expression of XHR1 was detected much earlier than that of other known MHB genes, XPax-2 and En-2, and also before the formation of the expression boundary between Xotx2 and Xgbx-2, suggesting that the early patterning of the presumptive MHB is independent of Xotx2 and Xgbx-2. Instead, the location of XHR1 expression appears to be determined in relation to the Xbra expression domain, since reduced or ectopic expression of Xbra altered the XHR1 expression domain according to the location of Xbra expression. In functional assays using mRNA injection, overexpression of dominant-negative forms of XHR1 in the MHB region led to marked reduction of XPax-2 and En-2 expression, and this phenotype was rescued by coexpression of wild-type XHR1. Furthermore, ectopically expressed wild-type XHR1 near the MHB region enhanced En-2 expression only in the MHB region but not in the region outside the MHB. These data suggest that XHR1 is required, but not sufficient by itself, to initiate MHB marker gene expression. Based on these data, we propose that XHR1 demarcates the prospective MHB region in the neuroectoderm in Xenopus early gastrulae. PMID- 11731237 TI - Scalloped and strawberry notch are target genes of Notch signaling in the context of wing margin formation in Drosophila. AB - The Notch pathway regulates the differentiation of many cell types throughout development of higher metazoa. Different cellular responses are elicited through specific activation of distinct Notch target genes. In the Drosophila wing, for example, the cut gene is activated by Notch signaling along the dorso-ventral boundary but, as we show here, not in other cell types. We identify additional regulatory components, scalloped and strawberry notch, that are targets of the Notch pathway specifically within the wing anlagen. As suggested by physical interactions, these proteins could be co-factors of the cut trans-regulator Vestigial. Additional regulatory input comes from the Wingless pathway. Our data support a model, whereby context specific involvement of distinct co-regulators modulates Notch target gene activation. PMID- 11731238 TI - Development and degeneration of dorsal root ganglia in the absence of the HMG domain transcription factor Sox10. AB - The HMG-domain transcription factor Sox10 is essential for the development of various neural crest derived lineages including glia and neurons of the peripheral nervous system (PNS). Within the PNS the most striking defect is the complete absence of glial differentiation whereas neurogenesis seemed initially normal. A degeneration of motoneurons and sensory neurons occurred later in development. The mechanism that leads to the dramatic effects on the neural crest derived cell lineages in the dorsal root ganglia (DRG), however, has not been examined up to now. Here, we provide a detailed analysis of proliferation and apoptosis in the DRG during the time of their generation and lineage segregation (between E 9.5 and E 11.5). We show that both increased apoptosis as well as decreased proliferation of neural crest cells contribute to the observed hypomorphism. PMID- 11731239 TI - A distant upstream region of the rat multipartite Na(+)-Ca(2+) exchanger NCX1 gene promoter is sufficient to confer cardiac-specific expression. AB - The Na(+)-Ca(2+) exchanger (NCX) regulates intracellular calcium homeostasis. We report on an upstream region of the rat NCX1 multipartite promoter that is active in cardiac myocytes. Although inactive in most non-cardiac cell lines, its activity can be rescued by cotransfection with GATA-4 and -6, but not GATA-5 transcription factors. In transgenic mice and similar to endogenous NCX1 mRNA expression, the upstream promoter region directs uniform beta-galactosidase expression in cardiac myocytes from approximately 7.75dpc. In adult mouse hearts, promoter activity is, however, significantly reduced and heterogeneous, except in the conduction system (sinoatrial and atrioventricular node, atrioventricular bundles). The upstream NCX1 promoter region thus directs appropriate spatial and temporal control of cardiac expression throughout development. PMID- 11731241 TI - Reduction of endogenous TGF-beta does not affect phenotypic development of sympathoadrenal progenitors into adrenal chromaffin cells. AB - Adrenal chromaffin cells and sympathetic neurons are related, but phenotypically distinct derivatives of the neural crest. Molecular cues that determine the chromaffin cell phenotype have not yet been identified; in contrast to a widely held belief, glucocorticoid signaling is apparently not relevant (Development 126 (1999) 2935). Transforming growth factor-betas (TGF-betas) regulate various aspects of embryonic development and are expressed in the environment of sympathoadrenal (SA) progenitor cells. We have previously shown that neutralization of endogenous TGF-beta from E4 to E8 in the quail embryo significantly increases numbers of adrenal tyrosine hydroxylase-positive cells. Whether endogenous TGF-beta may also be involved in influencing phenotypic development of adrenal chromaffin cells and their SA progenitors has not been analyzed. We now demonstrate that neutralization of endogenous TGF-beta1, -beta2 and -beta3 with a pan-anti-TGF-beta antibody in quail embryos during distinct time windows does not alter phenotypic development of chromaffin cells. In situ hybridizations revealed unaltered expression of neurofilament (NF-160), synaptotagmin I and neurexin I in adrenal glands. Likewise, the NF-associated antigen 3A10, and polyphosphorylated NF epitopes (RT 97) were unaltered. Most importantly, the typical ultrastructure of adrenal chromaffin cells including their large chromaffin secretory granules, a hallmark of the neuroendocrine phenotype, which distinguishes them from sympathetic neurons, was not affected. We therefore conclude that neutralization of endogenous TGF-beta influences chromaffin cell proliferation, but does not interfere with the development of the typical chromaffin cell phenotype. PMID- 11731240 TI - Conditional deletion of the bcl-x gene from mouse mammary epithelium results in accelerated apoptosis during involution but does not compromise cell function during lactation. AB - In the mammary gland Bcl-x is the most abundant cell survival factor from the Bcl 2 family. Since Bcl-x null mice die around day 12 of embryogenesis, the relevance of this protein in organ development and function is poorly understood. In erythroid cells bcl-x gene expression is controlled by cytokines and the transcription factor Stat5 (signal transducer and activator of transcription). However, we identified that bcl-x RNA levels in mammary tissue from prolactin receptor- and Stat5-null mice were indistinguishable from wild type mice. We have proposed that Bcl-x might control the survival of mammary epithelial cells throughout pregnancy, lactation, and the early stages of involution, and we have now tested this hypothesis through the conditional deletion of the bcl-x gene from mouse mammary epithelium. Conditional (floxed) bcl-x alleles were excised from alveolar cells during pregnancy using a Cre transgene under the control of the whey acidic protein gene promoter. Deletion of the bcl-x gene from the entire epithelial compartment (ducts and alveoli) was achieved by expressing Cre recombinase under control of the mouse mammary tumor virus long terminal repeat. The absence of Bcl-x did not compromise proliferation and differentiation of mammary ductal and alveolar epithelial cells in virgin mice and during pregnancy and lactation. However, epithelial cell death and tissue remodeling were accelerated in the bcl-x conditional knockout mice during the first stage of involution. Concomitant deletion of the bax gene did not significantly modify the Bcl-x phenotype. Our results suggest that Bcl-x is not essential during mammopoiesis, but is critical for controlled apoptosis during the first phase of involution. PMID- 11731242 TI - FGFs control the patterning of the inner ear but are not able to induce the full ear program. AB - FGF2 or FGF8 applied ectopically, close to the developing otic placode enhances transcription of a subset of ear marker genes such as Nkx5-1, SOHo1 and Pax2. Other ear expressed genes (Dlx5 and BMP4) are not up-regulated by FGFs. Ectopic FGFs lead to an increase in size of the vestibulo-cochlear ganglion. This phenotypic change is due to an increased recruitment of epithelial cells to the neuronal fate rather than to an enhanced proliferation. We also observed an induction of additional, vesicle-like structures upon ectopic FGF treatment, but this induction never led to enrolment of a full ear program. We further demonstrate that FGF8 is expressed in two separate, short waves, first at the otic placode stage and later at the vesicle stage. Both activities correspond to critical morphogenetic events in ear development. We propose that FGF8 is an important regulator of otocyst patterning. PMID- 11731243 TI - Vsx1, a rapidly evolving paired-like homeobox gene expressed in cone bipolar cells. AB - The paired-like homeodomain (HD) protein Chx10 is distinguished by the presence of the CVC domain, a conserved 56 amino acid sequence C-terminal to the HD. In mammals, Chx10 is essential both for the proliferation of retinal progenitor cells and for the formation or survival of retinal bipolar interneurons. We describe the cloning and characterization of a mouse Chx10 homologue, Vsx1; phylogenetic analysis suggests that Vsx1 and its putative vertebrate orthologues have evolved rapidly. Vsx1 expression in the adult is predominantly retinal. Whereas Chx10 is expressed both in retinal progenitors in the developing eye and apparently in all bipolar cells of the mature retina, Vsx1 expression is first detected in the eye at postnatal day 5, where it is restricted to cone bipolar cells. PMID- 11731244 TI - Emx2 in adult neural precursor cells. AB - Emx2 is a vertebrate homeobox gene involved in the control of the central nervous system development. In the formation of cerebral cortex, Emx2 expression is restricted mainly to the germinal ventricular zone fading away in the first postmitotic neurons. This expression pattern, the severe impairment of cortex organization and the size in mutant mice suggest a role of Emx2 in the control of proliferation and migration of neural precursor cells. The observed persistence of Emx2 expression in adult neurogenic areas in vivo is here confirmed at later stages. We also find that Emx2 is expressed at high levels in adult neural stem cells (ANSCs) in vitro and is down modulated upon differentiation. Overexpression of Emx2 gene in ANSCs has an anti-proliferative effect but it does not influence a particular differentiation pathway. Our results suggest that Emx2 may act promoting an asymmetric mode of cell division thereby increasing the size of a transit amplifying population. PMID- 11731245 TI - Temporally and spatially restricted expression of the helix-loop-helix transcriptional regulator Id1 during avian embryogenesis. AB - We isolated the chick orthologue of the Id1 helix-loop-helix gene and analyzed its expression pattern during early chick embryo development by whole-mount in situ hybridization. The Id1 expression pattern is dynamic and confined to discrete locations including the neural plate border, prospective olfactory placode, hindbrain, mesenchyme of distal branchial arches and adjacent to placodes, and the distal mesoderm of the limb buds. PMID- 11731246 TI - The transcriptional regulator Id3 is expressed in cranial sensory placodes during early avian embryonic development. AB - The chick homologue of the helix-loop-helix gene Id3 was isolated, and its expression pattern was analyzed during early stages of chick development. Chick Id3 is dynamically expressed in the olfactory, lens, and otic placodes. It is also observed in the epiphysis, nephric primordium, stomodeum, dermomyotome, distal branchial arches, dorsolateral hindbrain, foregut endoderm, dorsal spinal cord, and somites. PMID- 11731247 TI - Id4 expression and its relationship to other Id genes during avian embryonic development. AB - We present the sequence and expression pattern of chick Id4 and compare its distribution to that of other vertebrate Id genes. At early stages, Id4 expression is discrete, with transcript transiently expressed in subsets of migrating neural crest cells, the dorsal myocardium, the segmental plate mesoderm, and the tail bud. Later, expression is also observed in the telencephalic vesicles and corneal epithelium. Of all the Id genes, Id4 exhibits the most restricted pattern in the developing nervous system, with little expression in the presumptive neural crest or placodes. Id4 appears in the neural tube much later than other Id genes. However, all four Id genes display overlapping patterns in the branchial arches and tail bud. PMID- 11731248 TI - Expression pattern of two zebrafish genes, cxcr4a and cxcr4b. AB - We cloned and mapped two novel zebrafish genes, cxcr4a and cxcr4b, which are closely related to mammalian CXCR4. Expression analysis by reverse transcription polymerase chain reaction and in situ hybridization demonstrated that these two genes are expressed in most cell lineages known to express Cxcr4 in mammals. These genes are co-expressed in lateral mesoderm and posterior midbrain. The transcripts of cxcr4a were detected in interneurons and endoderm, whereas cxcr4b was specifically expressed in sensory neurons, motoneurons and cerebellum. In the lateral mesoderm, cxcr4b transcripts appeared earlier than those of cxcr4a. Thus, the function of mammalian CXCR4 could be split between the two zebrafish genes. These genes probably derived from the genome duplication event, which occurred during the evolution of teleosts. Similar pairs of Cxcr4 may exist in other species, where genome duplication has occurred. PMID- 11731249 TI - Notch pathway genes are expressed in mammalian ovarian follicles. AB - Folliculogenesis is the process of development of ovarian follicles that ultimately results in the release of fertilizable oocytes at ovulation. This is a complex program that involves the proliferation and differentiation of granulosa cells. Granulosa cells are necessary for follicle growth and support the oocyte during folliculogenesis. Genes that regulate the proliferation and differentiation of granulosa cells are beginning to be elucidated. In this study, the expression patterns of Notch receptor genes and their ligands, which have been shown to regulate cell-fate decisions in many systems during development, were examined in the mammalian ovary. In situ hybridization data showed that Notch2, Notch3, and Jagged2 were expressed in an overlapping pattern in the granulosa cells of developing follicles. Jagged1 was expressed in oocytes exclusively. Downstream target genes of Notch also were expressed in granulosa cells. These data implicate the Notch signaling pathway in the regulation of mammalian folliculogenesis. PMID- 11731250 TI - Expression of the Dan gene during chicken embryonic development. AB - The Dan gene was first identified as the putative rat tumor suppressor gene and encodes a protein structurally related to Cerberus and Gremlin in vertebrates. Xenopus DAN, as with Cerberus and Gremlin, was demonstrated to block bone morphogenetic protein (BMP) signaling by binding BMPs, and to be capable of inducing additional anterior structures by ectopic overexpression in Xenopus embryos. DAN, thus, is suggested to play pivotal roles in early patterning and subsequent organ development, as in the case of other BMP antagonists. In this report, we isolated the chicken counterpart of Dan. Chicken Dan is mainly expressed in the cephalic and somitic mesoderm and several placodes during organ development. PMID- 11731251 TI - Expression of Sprouty genes 1, 2 and 4 during mouse organogenesis. AB - We demonstrate that Sprouty genes 1, 2 and 4 are expressed in several developing organs of the craniofacial area and trunk, including the brain, cochlea, nasal organs, teeth, salivary gland, lungs, digestive tract, kidneys and limb buds. In organs such as the semicircular canal, Rathke's pouch, nasal organs, the follicle of vibrissae and teeth, Sprouty1 and Sprouty2 are expressed in the epithelium and Sprouty4 in the mesenchyme or neuronal tissue, while in the lung Sprouties1, 2 and 4 are all expressed mainly in the epithelial tissue. In the kidney, Sprouty1 is prominent in the ureteric bud whereas Sprouty2 and 4 are expressed in both the ureteric bud and the kidney mesenchyme and glomeruli deriving from it. The expression profiles suggest roles for these Sprouties in the epithelial mesenchymal interactions that govern organogenesis. PMID- 11731252 TI - Genome-wide analysis of Sox genes in Drosophila melanogaster. AB - Genes of the Sox family encode evolutionarily conserved HMG box containing transcription factors, which play key roles in various events of cell determination/differentiation during development. The total number of Sox genes in Drosophila melanogaster was estimated to be eight, after classical molecular cloning approaches and exhaustive screening of the complete Drosophila genome. Here we report the embryonic and larval expression pattern of four previously uncharacterized Sox genes, through antibody staining and in situ hybridization experiments. PMID- 11731253 TI - Cloning of HOXD1 from unfertilised human oocytes and expression analyses during murine oogenesis and embryogenesis. AB - We describe the cloning of HOXD1 in human unfertilised oocytes and detailed expression analyses during mouse oogenesis and embryogenesis. The cDNA of 1991bp has an open reading frame of 987bp encoding a protein of 329 amino acids. A comparison of the amino acid sequence with the mouse homologue revealed an overall homology of 85.5% with 99% identity within the homeodomain. Expression was detected in unfertilised human oocytes and 2-, 4-, 8-cell and blastocyst stage embryos. Expression analyses in mature mouse ovaries, early embryos and isolated gut revealed expression in the oocytes of the primary and secondary ovarian follicles, and in embryonal mesodermal derivatives such as dermatomes, urogenital tubercle, tail bud, kidney, ovaries, testes and enteric mesoderm adjacent to the caecum where expression was up-regulated in vitro in response to increasing doses of retinoic acid. Our observations indicate a possible role for HOXD1/Hoxd1 in the ovarian oocytes and the establishment of mesodermal derivatives during embryogenesis. PMID- 11731254 TI - Sequence and expression pattern of ziro7, a novel, divergent zebrafish iroquois homeobox gene. AB - We have isolated the zebrafish ziro7 gene, a novel, divergent member of the Iroquois family. ziro7 is expressed at early epiboly stages in the dorsal half of the zebrafish embryo, with a higher level in the dorso-lateral margin. From mid gastrulation stages onward, ziro7 is expressed in a large transversal stripe in the future neural plate, which subsequently divides into thinner stripes located in the diencephalon, midbrain and hindbrain. PMID- 11731255 TI - Sequential expression of annexin A5 in the vasculature and skeletal elements during mouse development. AB - Annexin A5 (annexin V, anchorin CII) represents the prototype member of the large annexin family, characterized by its ability to interact with phospholipids in a calcium-dependent manner and to form calcium-specific ion channels. Despite intense biochemical analysis, the in vivo expression and function of this annexin during mouse development, still remains unclear. Immunohistochemistry, in situ hybridization and reporter gene expression were used to define expression of annexin A5 during early mouse development. First, annexin A5 expression is associated with the developing vascular system. Later, expression is detected within the notochord and found in parallel to the differentiation of cartilage and bone. Therefore, expression of the Anxa5 gene may represent a novel marker characterizing cell lineages involved in the development of the vascular as well as the skeletal system. PMID- 11731256 TI - Place- and time-dependent expression of mouse sFRP-1 during development of the cerebral neocortex. AB - Wnts are a family of secreted proteins involved in multiple developmental mechanisms during nervous system development, including cell proliferation, cell migration, axon guidance and specification of cell positional information. We report here the expression of sFRP-1 mRNA, encoding a putative inhibitor of Wnt, in the developing mouse neocortex during the entire period when neurons for the neocortex are born. We show that sFRP-1 mRNA expression is spatially restricted to the proliferative zones during the period, when neurons are known to be generated in large numbers for the enlarging cortical plate. PMID- 11731257 TI - Expression of Deltex1 during mouse embryogenesis: comparison with Notch1, 2 and 3 expression. AB - The Notch signalling pathway defines a phylogenetically conserved cell-cell communication process that enables cell-fate specification in multicellular organisms. Deltex is a component of the Notch signalling network that physically interacts with the ankyrin repeats of Notch. Here, we report on the expression pattern of the Deltex1 gene during mouse embryonic development and, furthermore, we compare its expression with that of the Notch1, 2 and 3 genes. Complementary and combinatorial expression patterns between Deltex1 and the three Notch genes were observed throughout embryogenesis since Deltex1 expression was related either to cytodifferentiation (i.e. neuronal tissues) or to cell proliferation events (i.e. eye, vascular structures, hematopoiesis). PMID- 11731258 TI - Cloning and expression analysis of chicken Lix1, a founding member of a novel gene family. AB - Limb Expression 1 (Lix1), a founding member of a novel gene family, was identified in a screen for genes transiently and locally expressed during early chicken limb development. Most prominently, Lix1 is transiently expressed in the nascent hindlimb bud between Hamburger-Hamilton stages 15 and 19. Chicken Lix1 transcripts are also found in the basal plate of rhombomeres 3 and 5, in pharyngeal and in foregut mesenchyme and in all facial primordia except for the mandibular arches. Homologs of chick Lix1 exist in human, mouse and Drosophila. PMID- 11731259 TI - Expression of brain subtype creatine kinase in the zebrafish embryo. AB - Creatine kinases (CK) play crucial roles in intracellular energy transfer. We have isolated a cDNA from zebrafish embryos, which encodes a CK highly related to the mammalian brain subtype creatine kinase (BCK). The bck mRNA is expressed maternally in the zebrafish embryo and transcripts are distributed uniformly in blastula and gastrula stages. Expression becomes restricted to the prechordal plate and the nervous system during subsequent somitogenesis stages. bck transcripts are abundant in primary neurons in the developing central nervous system of the 1-day-old embryo. While some bck expression persists in the hindbrain, expression vanishes in the spinal cord of the 2-day-old embryo. In summary, the expression pattern of bck is highly dynamic and suggests a role for bck during gastrulation and neuronal differentiation. PMID- 11731260 TI - Spatial and temporal expression pattern of Runx3 (Aml2) and Runx1 (Aml1) indicates non-redundant functions during mouse embryogenesis. AB - The human RUNX3/AML2 gene belongs to the 'runt domain' family of transcription factors that act as gene expression regulators in major developmental pathways. Here, we describe the expression pattern of Runx3 during mouse embryogenesis compared to the expression pattern of Runx1. E10.5 and E14.5-E16.5 embryos were analyzed using both immunohistochemistry and beta-galactosidase activity of targeted Runx3 and Runx1 loci. We found that Runx3 expression overlapped with that of Runx1 in the hematopoietic system, whereas in sensory ganglia, epidermal appendages, and developing skeletal elements, their expression was confined to different compartments. These data provide new insights into the function of Runx3 and Runx1 in organogenesis and support the possibility that cross regulation between them plays a role in embryogenesis. PMID- 11731261 TI - Expression domains in the skin of genes affected by the nude mutation and identified by gene expression profiling. AB - Although several genes affected by the nude mutation inactivating the transcription factor Whn have recently been identified, a comprehensive molecular analysis of the nude phenotype is still missing. Gene expression profiling of wildtype and nude mice back skin reveals several so far unknown differences in mRNA levels and demonstrates that microarray hybridization is ideal to identify even quantitative changes in expression. Some genes are upregulated in the absence of Whn. Most of the differentially expressed genes are downregulated in nude skin. Our results identify metallothionein IV among these genes. This is the first report on metallothionein IV expression in the murine hair follicle; its expression domain almost completely overlaps that of Whn. PMID- 11731262 TI - Expression of receptor protein-tyrosine phosphatase alpha, sigma and LAR during development of the zebrafish embryo. AB - Receptor protein-tyrosine phosphatases (RPTPs) are key players in Drosophila development. To study the role of RPTPs in vertebrate development, we have cloned zebrafish (zf) RPTPs, including RPTP alpha (RPTPalpha), RPTP sigma (RPTPsigma) and LAR. These three RPTPs are broadly transcribed in early development. At 24h post fertilisation (hpf), all three genes are expressed in the nervous system in partially overlapping patterns. At 3 days post fertilisation zf-RPTPalpha and zf LAR show similar expression patterns in the central nervous system (CNS), the pharyngeal arches, the pectoral fins and the spinal cord. Interestingly, zf-LAR is uniquely expressed in the neuromast cells, whereas zf-RPTPsigma expression is confined to the central nervous system. PMID- 11731263 TI - Cloning and expression of the TALE superclass homeobox Meis2 gene during zebrafish embryonic development. AB - Meis and Prep/Pknox (MEINOX family) proteins, together with Pbx (PBC family) proteins, belong to the TALE superfamily characterized by an atypical homeodomain containing three additional amino acids between helix 1 and helix 2. Members of the MEINOX and PBC families have been isolated in Caenorhabditis elegans, Drosophila, Xenopus, chick, mouse and human, and play crucial roles in many aspects of embryogenesis. Here, we report the isolation of meis2 in zebrafish. Expression of meis2 is first detected at the beginning of gastrulation. Later during embryogenesis, meis2 transcripts are found in distinct domains of the central nervous system with the strongest expression in the hindbrain. Expression was also detected in the isthmus, along the spinal cord and in the lateral mesoderm. As development proceeds, meis2 is also expressed in the developing retina, pharyngeal arches, and in the vicinity of the gut tube. PMID- 11731264 TI - Tissue-specific expression of the Ets gene Xsap-1 during Xenopus laevis development. AB - We report the cloning of Xenopus laevis Xsap-1 cDNA, encoding a member of the ternary complex factor subfamily of ETS transcription factors. The expression pattern of Xsap-1 was examined during Xenopus embryogenesis using whole-mount in situ hybridization. Spatial expression of Xsap-1 mRNA is first detected at the animal pole at the mid-blastula stage. During neurulation Xsap-1 is expressed in cells participating in neural tube formation, in the sensorial layer of the epidermal ectoderm, and in an anterior region of the ventral mesoderm. Later, Xsap-1 expression is observed in the eye, ear vesicle, branchial arches, heart, pronephros, in the somites, and the developing nervous system, such as fore-, mid , and hindbrain as well as in the cranial ganglion X. PMID- 11731265 TI - Expression of Tcf/Lef and sFrp and localization of beta-catenin in the developing mouse lung. AB - Recent evidence that Wnts and other genes in the Wnt signaling pathway are expressed in embryonic and adult mouse lung suggests that this pathway is important for cell fate decisions and differentiation of lung cell types. We therefore examined the expression and protein distribution of several Wnt pathway components during prenatal mouse lung development using whole-mount in situ hybridization and immunohistochemistry. Between embryonic days 10.5 and 17.5 (E10.5-E17.5), beta-catenin was localized in the cytoplasm, and often also the nucleus, of the undifferentiated primordial epithelium (PE), differentiating alveolar epithelium (AE; present from E14.5 onward), and adjacent mesenchyme. Tcf1, Lef1, Tcf3, Tcf4, sFrp1, sFrp2 and sFrp4 were also expressed in the PE, AE, and adjacent mesenchyme in specific spatio-temporal patterns. PMID- 11731267 TI - The six hyaluronidase-like genes in the human and mouse genomes. AB - The human genome contains six hyaluronidase-like genes. Three genes (HYAL1, HYAL2 and HYAL3) are clustered on chromosome 3p21.3, and another two genes (HYAL4 and PH-20/SPAM1) and one expressed pseudogene (HYALP1) are similarly clustered on chromosome 7q31.3. The extensive homology between the different hyaluronidase genes suggests ancient gene duplication, followed by en masse block duplication, events that occurred before the emergence of modern mammals. Very recently we have found that the mouse genome also has six hyaluronidase-like genes that are also grouped into two clusters of three, in regions syntenic with the human genome. Surprisingly, the mouse ortholog of HYALP1 does not contain any mutations, and unlike its human counterpart may actually encode an active enzyme. Hyal-1 is the only hyaluronidase in mammalian plasma and urine, and is also found at high levels in major organs such as liver, kidney, spleen, and heart. A model is proposed suggesting that Hyal-2 and Hyal-1 are the major mammalian hyaluronidases in somatic tissues, and that they act in concert to degrade high molecular weight hyaluronan to the tetrasaccharide. Twenty-kDa hyaluronan fragments are generated at the cell surface in unique endocytic vesicles resulting from digestion by the glycosylphosphatidyl-inositol-anchored Hyal-2, transported intracellularly by an unknown process, and then further digested by Hyal-1. The two beta-exoglycosidases, beta-glucuronidase and beta-N-acetyl glucosaminidase, remove sugars from reducing termini of hyaluronan oligomers, and supplement the hyaluronidases in the catabolism of hyaluronan. PMID- 11731268 TI - Hyal2--less active, but more versatile? AB - Hyal2 is one of several hyaluronidases present in vertebrates. The human gene encoding this enzyme is present on chromosome 3p.21.3, close to two additional hyaluronidase genes. cDNAs encoding Hyal2 homologues have been characterized from mouse and Xenopus laevis. These enzymes hydrolyze high molecular mass hyaluronan to intermediates of approximately 20 kDa, a finding which implies that structural domains of this size exist in this polysaccharide which was mostly thought to be a random coil. Hyal2 enzymes have an acidic pH-optimum with an activity that is considerably lower than observed for other types of hyaluronidases. Originally considered to be a typical lysosomal enzyme, more recent evidence has shown that Hyal2 proteins can also be exposed on the cell surface bound to the plasma membrane via a GPI anchor. Hyal2 is present in many tissues, one exception being the adult brain. In this tissue, the gene is silenced after birth by methylation. Current evidence about the role of Hyal2 in tumor growth, inflammation and frog embryogenesis is discussed. PMID- 11731269 TI - The dual functions of GPI-anchored PH-20: hyaluronidase and intracellular signaling. AB - The ovulated mammalian oocyte is surrounded by the "cumulus ECM", composed of cells embedded in an extracellular matrix that is rich in hyaluronic acid (HA). The cumulus ECM is a viscoelastic gel that sperm must traverse prior to fertilization. Mammalian sperm have a GPI-anchored hyaluronidase which is known as PH-20 and also as SPAM 1. PH-20 is located on the sperm surface, and in the lysosome-derived acrosome, where it is bound to the inner acrosomal membrane. PH 20 appears to be a multifunctional protein; it is a hyaluronidase, a receptor for HA-induced cell signaling, and a receptor for the zona pellucida surrounding the oocyte. The zona pellucida recognition function of PH-20 was discovered first. This function is ascribed to the inner acrosomal membrane PH-20, which appears to differ biochemically from the PH-20 on the sperm surface. Later, when bee venom hyaluronidase was cloned, a marked cDNA sequence homology with PH-20 was recognized, and it is now apparent that PH-20 is the hyaluronidase of mammalian sperm. PH-20 is unique among the hyaluronidases in that it has enzyme activity at both acid and neutral pH, and these activities appear to involve two different domains in the protein. The neutral enzyme activity of plasma membrane PH-20 is responsible for local degradation of the cumulus ECM during sperm penetration. Plasma membrane PH-20 mediates HA-induced sperm signaling via a HA binding domain that is separate from the hyaluronidase domains. This signaling is associated with an increase in intracellular calcium and as a consequence, the responsiveness of sperm to induction of the acrosome reaction by the zona pellucida is increased. There is extensive evidence that GPI-anchored proteins are involved in signal transduction initiated by a diverse group of cell surface receptors. GPI-anchored proteins involved in signaling are often associated with signaling proteins bound to the cytoplasmic leaflet of the plasma membrane, typically Src family, non-receptor protein tyrosine kinases. PH-20 appears to initiate intracellular signaling by aggregating in the plasma membrane, and a 92 kDa protein may be the cell signaling molecule linked to PH-20. PMID- 11731270 TI - A member of the Y-box protein family interacts with an upstream element in the alpha1(I) collagen gene. AB - Transforming growth factor beta (TGF-beta) stimulates protein complex formation on a TGF-beta response element (TAE) found in the distal portion (-1624) of the collagen alpha 1(I) promoter. To identify the fibroblast proteins in this complex, an expression library constructed from human embryonic lung fibroblasts mRNA was screened using a tetramer of TAE. Y-box binding protein (YB-1), was identified as a protein in the TAE-protein complex. The protein expressed by phage clones formed a specific complex with labeled TAE but not mutated TAE (mTAE) similar to the complex formed with nuclear protein. Nuclear protein-TAE complexes isolated from native gels contained YB-1 by Western analysis. TGF-beta treatment increased the amount of YB-1 protein in nuclear extracts, decreased its amount in cytoplasm, but did not alter the steady state levels of YB-1 mRNA. A full-length YB-1 protein expressed in human lung fibroblasts was primarily located in the nucleus with punctate staining in cytoplasmic regions. The expression of YB-1 decreased in the cytoplasm after 2 h of TGF-beta treatment. Therefore, the increased binding activity seen in TGF-beta-stimulated nuclear extracts was due primarily to relocalization of YB-1 from the cytoplasm to the nuclear compartment. Co-transfection of YB-1 cDNA with a collagen promoter reporter construct caused a dose-dependent activation of collagen promoter activity in rat fibroblasts whereas the promoter with a mutation in the TAE element was not sensitive to YB-1 co-expression. In conclusion, we have identified YB-1 as a protein that interacts with a TGF-beta response element in the distal region of the collagen alpha 1(I) gene. YB-1 protein activates the collagen promoter and translocates into the nucleus during TGF-beta addition to fibroblasts, suggesting a role for this protein in TGF-beta signaling. PMID- 11731271 TI - Cathepsin D cleaves aggrecan at unique sites within the interglobular domain and chondroitin sulfate attachment regions that are also cleaved when cartilage is maintained at acid pH. AB - Bovine aggrecan was digested with bovine cathepsin D at pH 5.2 under conditions of partial digestion and the resulting aggrecan core protein fragments were separated by electrophoresis on gradient polyacrylamide gels. The fragments were characterized by their reactivity to specific antibodies and by N-terminal amino acid sequencing. It was also demonstrated that cathepsin D cleaved bovine aggrecan at five sites within the core protein, between residues Phe(342) Phe(343) in the interglobular domain, Leu(1462)-Val(1463) between the chondroitin sulfate attachment regions 1 and 2 and Leu(1654)-Val(1655), Phe(1754)-Val(1755) and Leu(1854)-Ile(1855) that are located within the chondroitin sulfate attachment region 2 of the core protein. The time course of digestion showed that there was a continued degradation of aggrecan and there was no preferential cleavage of the core protein at any one site. It was shown that cathepsin D digested aggrecan over the pH range 5.2-6.5 resulting in the same products. When bovine cartilage was maintained in explant culture at pH 5.2 there was a rapid loss of both radiolabeled and chemical pools of sulfated glycosaminoglycans into the culture medium and this loss was inhibited by the inclusion in the medium of the aspartic proteinase inhibitor, pepstatin A. The aggrecan core protein fragments appearing in the medium of cultures maintained at pH 5.2 were characterized and it was shown that the fragments had N-terminal sequences starting at Phe(343), Ile(1855), and Val(1755) or Val(1463). This work demonstrates that cathepsin D present within the extracellular matrix of articular cartilage has the potential to contribute to the proteolytic processing of the core protein of aggrecan in this tissue. PMID- 11731272 TI - Characterization and expression of murine PRELP. AB - The cDNA sequence of the murine proline/arginine-rich end leucine-rich repeat protein (PRELP) gene was cloned by PCR-based techniques. The gene encodes a protein of 378 amino acids, which is four amino acid residues shorter than its human counterpart. This difference resides mainly in the amino terminal region of the mature protein, which is five amino acids shorter in the mouse than the human and has a lower arginine content. The remainder of the protein, including the structure of the leucine-rich repeats, the potential sites for N-linked glycosylation, and the disulfide-bonded domains are well conserved between species. In common with humans, the murine gene possesses three exons, with the translation initiation codon residing in exon 2 and the termination codon in exon 3. Exons 1 and 2 are separated by an intron of approximately 6.7 kbp, whereas exons 2 and 3 are separated by an intron of approximately 1.7 kbp. Western blot analysis of mouse cartilage extracts indicates that PRELP exists as a glycoprotein of approximately 55 kDa, as in human cartilage. Immunohistochemical and in situ hybridization analysis reveal that PRELP is expressed in cartilage throughout both fetal development and post-natal life, in contrast to the human where expression in cartilage is not apparent prior to birth. Northern blot analysis indicates that PRELP mRNA is also expressed in the developing embryo prior to skeletogenesis. The promoter region of the mouse PRELP gene possesses no TATA box in its proximal region, in common with humans, and shows differences in the conservation of elements known to be involved in regulating expression of the human PRELP gene. PMID- 11731273 TI - Characterization of the mouse integrin subunit alpha10 gene and comparison with its human homologue. Genomic structure, chromosomal localization and identification of splice variants. AB - Alpha10beta1 is a collagen-binding integrin expressed by chondrocytes [Camper et al. (1998) J. Biol. Chem. 273: 20383-20389]. In the present study, the mouse alpha10 gene was isolated from a sCos1 SVJ library and the genomic structure and chromosomal localization was determined. The alpha10 gene consists of 30 translated exons spanning a region of approximately 18 kb genomic DNA. The sequences of all exon/intron borders follow the consensus "gt-ag" rule. A transcription start site, determined by primer extension analysis, was located 38 nucleotides upstream of the initiation ATG site. The 5' flanking region of the transcription start site lacked a TATA-box. The first exon contained, in addition to 38 untranslated nucleotides, the ATG translation start site and the major part of the signal peptide. The alpha10 gene was mapped to mouse chromosome 3 by fluorescence in situ hybridization and is the only integrin subunit localized to this chromosome. When we investigated the expression of alpha10 by PCR we found that both mouse and human articular chondrocytes express extracellular splice variants of the alpha10 subunit. In mouse, exon 26 was extended into the intron by 62 nt, generating a truncated alpha10-chain. In human, exon 25 consisted of 114 nt which were alternately spliced in or out. PMID- 11731274 TI - Neutrophil collagenase (MMP-8) is expressed during early development in neural crest cells as well as in adult melanoma cells. AB - Matrix metalloproteinase-8 (MMP-8) is a neutral metalloproteinase of the fibrillar collagenase family that also includes MMP-1 and MMP-13. In contrast to the other collagenases, MMP-8 has a very limited tissue distribution, thought to be restricted to neutrophils and chondrocytes. In a previous study, we observed MMP-8 expression in human melanoma cells. This observation led us to assess in more detail the expression of MMP-8 in normal and malignant melanocytic cells. We found that MMP-8 was expressed by 11 out of 12 human melanoma cell lines tested and all 10 primary melanomas we examined, but was not expressed by four primary neonatal melanocyte strains. Since melanocytes arise from highly motile neural crest cells, we examined the hypothesis that MMP-8 might be expressed by neural crest cells. RT-PCR analysis of post-implantation mouse embryos indicated the presence of MMP-8 transcripts at E9.5. In situ hybridization and immunohistochemistry of mouse embryos between E9.5-E14.5 demonstrated MMP-8 expression in the surface ectoderm, neural crest cells and chondrocytes. MMP-8 was also detected in neural crest cell migration located in the circumference of the neural tube, branchial arches and the notochord. In addition, MMP-8 expression was observed between the somites, in circumscriptive areas of the developing brain, heart, and eye, and in the interdigital zones of the limbs. In summary, we found MMP-8 to be the first fibrillar collagenase expressed during development. In contrast to its restricted tissue expression post-partum, MMP-8 was present in multiple embryonic tissues, including neural crest cells. The production of MMP-8 by migrating neural crest cells may contribute to their ability to degrade fibrillar collagen matrices while in transit. PMID- 11731275 TI - Cis-acting elements regulate alternative splicing of exons 6A, 6B and 8 of the alpha1(XI) collagen gene and contribute to the regional diversification of collagen XI matrices. AB - Consecutive exons 6A, 6B, 7 and 8 that encode the variable region of the amino terminal domain (NTD) of the col11a1 gene product undergo a complex pattern of alternative splicing that is both tissue-dependent and developmentally regulated. Expression of col11a1 is predominantly associated with cartilage where it plays a critical role in skeletal development. At least five splice-forms (6B-7-8, 6A-7 8, 7-8, 6B-7 and 7) are found in cartilage. Splice-forms containing exon 6B or 8 have distinct distributions in the long bone during development, while in non cartilage tissues, splice-form 6A-7-8 is typically expressed. In order to study this complex and tissue-specific alternative splicing, a mini-gene that contains mouse genomic sequence from exon 5 to 11, flanking the variable region of alpha1(XI)-NTD, was constructed. The minigene was transfected into chondrocytic (RCS) and non-chondrocytic (A204) cell lines that endogenously express alpha1(XI), as well as 293 cells which do not express alpha1(XI). Alternative splicing in RCS and A204 cells reflected the appropriate cartilage and non cartilage patterns while 293 cells produced only 6A-7-8. This suggests that 6A-7 8 is the default splicing pathway and that cell or tissue-specific trans-acting factors are required to obtain pattern of the alternative splicing of alpha1(XI) pre-mRNA observed in chondrocytes. Deletional analysis was used to identify cis acting regions important for regulating splicing. The presence of the intact exon 7 was required to generate the full complex chondrocytic pattern of splicing. Furthermore, deletional mapping of exon 6B identified sequences required for expression of exon 6B in RCS cells and these may correspond to purine-rich (ESE) and AC-rich (ACE) exonic splicing enhancers. PMID- 11731276 TI - Tapping-mode atomic force microscopy in fluid of hydrated extracellular matrix. AB - Fragments of native, hydrated rat tail tendon were imaged by tapping-mode atomic force microscopy while immersed in fluid. The specimens were soft and sensitive to the operating parameters, and with minimal imaging pressure the collagen fibrils appeared covered by irregular blobs or by filamentous material. A slight increase in pressure caused the underlying fibril surface to appear, with an evident D-period, gap- and overlap-zones and three intraperiod ridges. Fibrils often ran parallel and in phase, implying some coupling mechanism. Longitudinal subfibrils, 8-9 nm thick, occasionally appeared. The simultaneous acquisition of the "tapping amplitude" along with the usual "height" channel clearly confirmed the presence of longitudinal subfibrils, indicative of the inner architecture of the fibril. PMID- 11731278 TI - Myopathy with lobulated muscle fibers: evidence for heterogeneous etiology and clinical presentation. AB - The clinico-pathological features of 17 patients displaying a myopathy with lobulated (trabeculated) fibers are reported. All these patients had a limb girdle phenotype and at least 20% of lobulated fibers in their muscle biopsies. There were ten females and seven males. The onset of symptoms ranged from 2 to 55 years (mean 24). The average age at the time of muscle biopsy was 39 (range 3 63). Interestingly, in six patients, high prevalence of lobulated fibers was observed at the second biopsy only, performed on average 11 years after the first or in another muscle. Six patients had a suggestively positive family history. Facial weakness was noted in two patients (genetic study confirmed FSH dystrophy). The course and the severity of weakness varied from one patient to another. Immunohistochemistry and Western blot analyses revealed one Duchenne carrier, one alpha-sarcoglycanopathy, no dysferlinopathy and four calpain deficiencies (including one patient with FSH dystrophy), but SSCP revealed mutation in the calpain gene in only one of the patients. These results show that (1) myopathies with lobulated fibers are clinically and genetically heterogeneous, (2) lack of calpain expression by Western blot analysis is not always associated with null mutation, (3) a molecular diagnosis is made in less than 40% of myopathy with lobulated fibers, (4) when observed, lobulated fibers are most prominent in proximal muscles and require time to appear. PMID- 11731279 TI - A new phenotype of autosomal dominant nemaline myopathy. AB - We present a five-generation family with a novel phenotype of autosomal dominant nemaline myopathy not linked to the three genes known to be causative for nemaline myopathy (alpha-tropomyosin-3, nebulin, and alpha-actin). Although there was muscle weakness in the neck flexors and proximal muscles of the limbs, as found in other families, facial, ankle dorsiflexor and respiratory muscles were normal. The most remarkable clinical feature was a peculiar kind of slowness in movement not reported previously in nemaline myopathy. PMID- 11731280 TI - Autosomal dominant Emery-Dreifuss muscular dystrophy: a new family with late diagnosis. AB - Emery-Dreifuss muscular dystrophy is characterized by the clinical triad of early onset contractures of elbows, Achilles tendons and spine, wasting and weakness with a predominantly humero-peroneal distribution and life-threatening cardiac conduction defects and/or cardiomyopathy. Two main types of inheritance have been described: the X-linked form is caused by mutations in the STA gene on locus Xq28 and the gene for the autosomal dominant form (LMNA gene) has been localized on chromosome 1q11-q23. Recently, mutations in this LMNA gene have been also found to be responsible for the less frequent autosomal recessive form of the disease. Although all forms share a similar clinical presentation, some differences appear to exist between them as has been described recently in a large number of patients. We present the first documented Spanish family genetically confirmed to have autosomal dominant Emery-Dreifuss muscular dystrophy. Clinical, pathological and genetic data are described. We emphasize the difficulties in diagnosis, especially in sporadic cases or young patients in whom the clinical picture is not completely established. PMID- 11731281 TI - Autosomal dominant distal spinal muscular atrophy: an Italian family not linked to 12q24 and 7p14. AB - Distal spinal muscular atrophy is genetically heterogeneous, as sporadic cases and both autosomal dominant and recessive inheritance have been described. An autosomal dominant distal spinal muscular atrophy with upper limb predominance has been mapped to chromosome 7p, and more recently, an autosomal dominant distal spinal muscular atrophy with lower limb predominance has been linked to chromosome 12q24. We describe a four generation Italian family with autosomal dominant distal spinal muscular atrophy starting between 8 and 30 years with weakness and atrophy of distal leg muscles. The older patients also presented sensorineural deafness. We performed genetic linkage analysis with microsatellite markers D12S366, D12S349, D12S86, D12S321, D12S1612, D12S1349, D12S342, PLA2A on chromosome 12q24 and D7S516, D7S2496, D7S632, D7S2252 on chromosome 7p14. No support for linkage to chromosome 12q24 and 7p14 was found in our family, confirming a genetic heterogeneity within autosomal dominant distal spinal muscular atrophy. PMID- 11731282 TI - Intolerance to neuroleptics and susceptibility for malignant hyperthermia in a patient with proximal myotonic myopathy (PROMM) and schizophrenia. AB - We report a patient with proximal myotonic myopathy who was treated with neuroleptics because of exacerbating schizophrenia. Under therapy with fluanxol, the patient developed muscle stiffness and oculogyric cramps. Treatment with both amisulpride and olanzapine lead to markedly elevated serum creatine kinase levels. An in-vitro contracture test was positive for halothane. Thus, in patients with all kinds of multisystemic myotonic myopathies, a susceptibility for malignant hyperthermia and intolerance towards neuroleptics should be taken into account. PMID- 11731283 TI - Expression of dystrophin-associated proteins during neuronal differentiation of P19 embryonal carcinoma cells. AB - The dystrophin gene that is defective in Duchenne muscular dystrophy shows a complex transcriptional control based on several promoters driving independent cell-type-specific expression of different isoforms. Dystrophin isoforms together with dystroglycan, a transmembrane protein which in turn binds to extracellular matrix, are the core of a complex of proteins, the dystrophin-associated protein (DAP) complex, which also comprises cytoplasmic elements like dystrobrevin. Whereas the molecular organization of DAP complex in muscle is well documented, the composition of a similar complex in the nervous system remains largely unknown. We followed by competitive PCR the expression of DAP complex components during retinoic acid (RA)-induced neuronal differentiation of P19 cells. Transcripts for the full-length dystrophin, Dp427, and the short isoform, Dp71, as well as for alpha-dystrobrevin 2 increased in parallel with days in culture after RA stimulation, while dystroglycan, alpha-dystrobrevin 1 and 3, and beta dystrobrevin were constitutively expressed. The upregulation of some of the components of the dystrophin complex during neuronal maturation suggests functional flexibility of the complex in the nervous system, where specific associations between different isoforms of DAP complex components could possibly organize distinct DAP complex-like complexes. PMID- 11731284 TI - A novel nonsense mutation (Q352X) in the mitochondrial cytochrome b gene associated with a combined deficiency of complexes I and III. AB - We identified a novel mitochondrial cytochrome b mutation in a patient with progressive exercise intolerance, muscle cramps and lactic acidosis. A marked reduction of the enzymatic activities of respiratory chain complexes I and III was found in muscle biopsy. The mutation was a heteroplasmic C15800T transition, determining a stop-codon at amino acid position 352 (Q352X). Mutant mtDNA was approximately 45% of total genomes in muscle, while it was absent in all of the other examined tissues of the patient and in lymphocytes of the patient's mother. Clinical presentation and laboratory findings strongly support the hypothesis that this mutation is the primary cause of the disease in our patient. PMID- 11731285 TI - A point mutation of mitochondrial ATPase 6 gene in Leigh syndrome. AB - A T-to-G transition at nucleotide 9176 (T9176G) in the mitochondrial adenosine triphosphate 6 gene (MTATP6) was detected in two siblings with Leigh syndrome. Heteroplasmy was observed in the mother's leukocytes. The T9176G mutation changes a highly conserved leucine residue to an arginine in subunit 6 of ATPase and is maternally inherited like mutations in the other mitochondrial genes. Another mutation in the same codon (T9176C) has been previously reported in Leigh syndrome. This gives strong support to the relevance of MTATP6 dysfunction in Leigh syndrome and the importance of leucine at that position. PMID- 11731286 TI - Hypertrophic cardiomyopathy and mtDNA depletion. Successful treatment with heart transplantation. AB - Cardiomyopathy associated with a mitochondrial DNA depletion syndrome is a rare condition. We report on a child with a hypertrophic cardiomyopathy and a mitochondrial depletion syndrome who was successfully treated by heart transplantation, given the tissue-specific nature of her mitochondrial disorder. PMID- 11731287 TI - 80th ENMC International Workshop on Multi-Minicore Disease: 1st International MmD Workshop. 12-13th May, 2000, Soestduinen, The Netherlands. PMID- 11731288 TI - 85th ENMC International Workshop on Congenital Muscular Dystrophy. 6th International CMD Workshop. 1st Workshop of the Myo-Cluster Project 'GENRE'. 27 28th October 2000, Naarden, The Netherlands. PMID- 11731293 TI - Synthesis and biological evaluation of all A-ring stereoisomers of 5,6-trans-2 methyl-1,25-dihydroxyvitamin D(3) and their 20-epimers: possible binding modes of potent A-ring analogues to vitamin D receptor. AB - BACKGROUND: The secosteroid 1 alpha,25-dihydroxyvitamin D(3) (1) has a wide variety of biological activities, which makes it a promising therapeutic agent for the treatment of cancer, psoriasis and osteoporosis. Insight into the structure-activity relationships of the A-ring of 1 is still needed to assist the development of more potent and selective analogues as candidate chemotherapeutic agents, as well as to define the molecular mode of action. RESULTS: All possible A-ring stereoisomers of 5,6-trans-2-methyl-1,25-dihydroxyvitamin D(3) (6a-h) and their 20-epimers (7a-h) were designed and efficiently synthesized. The dependence of the affinities for vitamin D receptor (VDR) and vitamin D binding protein (DBP), as well as the HL-60 cell differentiation-inducing activity, upon the stereochemistry of the A-ring and at C20 in the side chain was evaluated. CONCLUSIONS: The binding affinities and potency of the 5,6-trans and 5,6-cis analogues were enhanced by a 2-methyl substituent in a certain orientation. Molecular docking studies based upon the X-ray crystal structure of VDR suggested that the axial 2-methyl group would be accommodated in a pocket surrounded by hydrophobic amino acid residues in the ligand binding domain, resulting in enhanced interaction. PMID- 11731294 TI - De novo design of fibrils made of short alpha-helical coiled coil peptides. AB - BACKGROUND: The alpha-helical coiled coil structures formed by 25-50 residues long peptides are recognized as one of Nature's favorite ways of creating an oligomerization motif. Known de novo designed and natural coiled coils use the lateral dimension for oligomerization but not the axial one. Previous attempts to design alpha-helical peptides with a potential for axial growth led to fibrous aggregates which have an unexpectedly big and irregular thickness. These facts encouraged us to design a coiled coil peptide which self-assembles into soluble oligomers with a fixed lateral dimension and whose alpha-helices associate in a staggered manner and trigger axial growth of the coiled coil. Designing the coiled coil with a large number of subunits, we also pursue the practical goal of obtaining a valuable scaffold for the construction of multivalent fusion proteins. RESULTS: The designed 34-residue peptide self-assembles into long fibrils at slightly acid pH and into spherical aggregates at neutral pH. The fibrillogenesis is completely reversible upon pH change. The fibrils were characterized using circular dichroism spectroscopy, sedimentation diffusion, electron microscopy, differential scanning calorimetry and X-ray fiber diffraction. The peptide was deliberately engineered to adopt the structure of a five-stranded coiled coil rope with adjacent alpha-helices, staggered along the fibril axis. As shown experimentally, the most likely structure matches the predicted five-stranded arrangement. CONCLUSIONS: The fact that the peptide assembles in an expected fibril arrangement demonstrates the credibility of our conception of design. The discovery of a short peptide with fibril-forming ability and stimulus-sensitive behavior opens new opportunities for a number of applications. PMID- 11731295 TI - The inefficiency of incisions of ecteinascidin 743-DNA adducts by the UvrABC nuclease and the unique structural feature of the DNA adducts can be used to explain the repair-dependent toxicities of this antitumor agent. AB - BACKGROUND: Ecteinascidin 743 (Et 743), a natural product derived from a marine tunicate, is a potent antitumor agent presently in phase II clinical trials. Et 743 binds in the minor groove of DNA and alkylates N2 of guanine via a unique mechanism involving catalytic activation. The sequence selectivity of Et 743 is governed by different patterns of hydrogen-bonding to DNA, which results in differential reversibility of the covalent adducts. As determined by nuclear magnetic resonance spectroscopy, the preferred sequences 5'-PuGC and 5'-PyGG are stabilized by a hydrogen-bonding network, while the non-preferred sequences 5' NG(A/T) are much less stabilized due to the lack of a key hydrogen bond to the GC base pair on the 3'-side of the alkylated guanine. RESULTS: Mammalian cell lines (XPB, XPD, XPF, XPG, and ERCC1) deficient in the nucleotide excision repair (NER) gene products show resistance to Et 743. The recognition and subsequent incision of Et 743-DNA adducts by the bacterial multisubunit endonuclease UvrABC were used to evaluate DNA repair-mediated toxicity as a rationale for the resistance of NER defective cell lines and the antitumor activity of Et 743. The Et 743-DNA adducts are indeed recognized and incised by the UvrABC repair proteins; however, the pattern of incision indicated that the non-preferred, and less stable, sequences (i.e. 5'-NG(A/T)) modified with Et 743 are generally incised at a much higher efficiency than the preferred, more stable sequences (i.e. 5'-PuGC or 5'-PyGG). In addition, within the same Et 743 recognition sequence, the level of incision varies, indicating that flanking regions also contribute to the differential incision frequency. CONCLUSIONS: The inefficient repair incision by the UvrABC nuclease of Et 743-DNA adducts provides a basis for rationalizing the observed repair-dependent cytotoxicities of these DNA adducts, if other associated structural properties of Et 743-DNA adducts are taken into account. In particular, the wedge-shaped Et 743, which forces open the minor groove of DNA, introducing a major groove bend, and the extrahelical protrusion of the C-subunit of Et 743 provide unique characteristics alongside the hydrogen-bonding stabilization of a covalent DNA adduct, which we propose traps an intermediate in NER processing of Et 743-DNA adducts. This trapped intermediate protein-Et 743 DNA adduct complex can be considered analogous to a poisoned topoisomerase I- or topoisomerase II-DNA complex. In the absence of an intact NER nuclease complex, this toxic lesion is unable to form, and the Et 743-DNA adducts, although not repaired by the NER pathway, are less toxic to cells. Conversely, elevated levels of either of these nucleases should lead to enhanced Et 743 toxicity. PMID- 11731296 TI - A novel fluorescent silica tracer for biological silicification studies. AB - BACKGROUND: Biological silica production has drawn intense attention and several molecules involved in biosilicification have been identified. Cellular mechanisms, however, remain unknown mainly due to the lack of probes required for obtaining information on live specimens. RESULTS: The fluorescence spectra of the compound 2-(4-pyridyl)-5-((4-(2 dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole (PDMPO) are affected by the presence of >3.2 mM silicic acid. Increase in intensity and shift in the fluorescence coincide with the polymerization of Si. The unique PDMPO-silica fluorescence is explored here to visualize Si deposition in living diatoms. The fluorophore is selectively incorporated and co-deposited with Si into the newly synthesized frustules (the outer silica shells) showing an intense green fluorescence. CONCLUSIONS: We suggest that a fluorescence shift is due to an interaction between PDMPO and polymeric silicic acid. PDMPO is an excellent probe for imaging newly deposited silica in living cells and has also a potential for a wide range of applications in various Si-related disciplines, including biology of living organisms as diatoms, sponges, and higher plants, clinical research (e.g. lung fibrosis and cancer, bone development, artificial bone implantation), and chemistry and physics of materials research. PMID- 11731297 TI - Combinatorial library of five-membered iminocyclitol and the inhibitory activities against glyco-enzymes. AB - BACKGROUND: Oligosaccharide processing enzymes are important classes of catalysts involved in synthesizing specific oligosaccharide structures on proteins and sphingolipids. Development of specific inhibitors of such enzymes is of current interest as these inhibitors may be used to control cellular functions. Five membered iminocyclitols have been shown to be potent inhibitors of such enzymes. Since a rational design and synthesis of inhibitors is often extremely difficult due to the limited information regarding the structure of the active site, we carried out a combinatorial library approach. RESULTS: To create diversity, we decided to use an aldehyde group of a protected iminocyclitol for reductive amination and the Strecker reaction. After transformation of the nitrile group introduced by the Strecker reaction into an amine and amide and complete deprotection, a small library of five-membered iminocyclitols consisting of 27 compounds was synthesized. A series of compounds obtained by reductive amination was first screened as potential inhibitors of glycosidases and glycosyltransferases. Among them, compounds carrying a C(10)-alkyl group showed marked enhancement of inhibitory activity against alpha-mannosidase at 10 microM concentration when compared with its parent compound and deoxymannojirimycin. Furthermore, compounds having the phenylethyl group showed an extremely strong inhibitory effect against alpha-galactosaminidase at a K(i) value of 29.4 nM. Compounds with an aminomethyl and amide group at the C-1' position of these two molecules showed a decrease in inhibitory activities. CONCLUSIONS: A combinatorial approach based on five-membered iminocyclitols with a galacto configuration was exploited. The potential usefulness of the library as a source of inhibitors of glycoenzymes is clearly shown in this study. PMID- 11731298 TI - Intramolecular ditryptophan crosslinks enforce two types of antiparallel beta structures. AB - BACKGROUND: Two types of biaryl crosslinks can be formed with natural protein sidechains: ditryptophan and dityrosine. Biaryl crosslinks have the same topology as disulfide crosslinks, yet little is known about their effect on local peptide structure. RESULTS: Three ditryptophan-linked peptide dimers based on the sequence Ac-Leu-Trp-Ala-COX were prepared. The tripeptide dimer with -CONH(2) termini was too insoluble to study, but the tripeptide dimer with -COOMe termini crystallized from methanol/chloroform as an antiparallel beta-sheet. The tripeptide dimer with a -CONMe(2) termini adopted a slipped antiparallel beta structure in methanol/chloroform. CONCLUSIONS: These results suggest that intermolecular ditryptophan crosslinks that join the middle of peptide chains can confer a preference for antiparallel beta-sheet structure. The effect is most dramatic when both the inside and outside edges of the dimer can form hydrogen bonds as in the crystal structure of dimer 3b. PMID- 11731299 TI - Peptides that mimic glycosaminoglycans: high-affinity ligands for a hyaluronan binding domain. AB - BACKGROUND: Hyaluronan (HA) is a non-sulfated glycosaminoglycan (GAG) that promotes motility, adhesion, and proliferation in mammalian cells, as mediated by cell-surface HA receptors. We sought to identify non-carbohydrate ligands that would bind to and activate cell-surface HA receptors. Such analogs could have important therapeutic uses in the treatment of cancer, wound healing, and arthritis, since such ligands would be resistant to degradation by hyaluronidase (HAse). RESULTS: Peptide ligands that bind specifically to the recombinant HA binding domain (BD) of the receptor for hyaluronan-mediated motility (RHAMM) were obtained by screening two peptide libraries: (i) random 8-mers and (ii) biased 8 mers with alternating acidic side chains, i.e. XZXZXZXZ (X=all-L-amino acids except Cys, Lys, or Arg; Z=D-Asp, L-Asp, D-Glu, or L-Glu). Selectivity of the peptide ligands for the HABD was established by (i) detection of binding of biotin- or fluorescein-labeled peptides to immobilized proteins and (ii) fluorescence polarization of FITC-labeled peptides with the HABD in solution. HA competitively displaced binding of peptides to the HABD, while other GAGs were less effective competitors. The stereochemistry of four biased octapeptides was established by synthesis of the 16 stereoisomers of each peptide. Binding assays demonstrated a strong preference for alternating D and L configurations for the acidic residues, consistent with the calculated orientation of glucuronic acid moieties of HA. CONCLUSIONS: Two classes of HAse-resistant peptide mimetics of HA were identified with high affinity, HA-compatible binding to the RHAMM HABD. This demonstrated that non-HA ligands specific to a given HA binding protein could be engineered, permitting receptor-specific targeting. PMID- 11731300 TI - Direct interaction of a vancomycin derivative with bacterial enzymes involved in cell wall biosynthesis. AB - BACKGROUND: The glycopeptide antibiotic vancomycin complexes DAla-DAla termini of bacterial cell walls and peptidoglycan precursors and interferes with enzymes involved in murein biosynthesis. Semisynthetic vancomycins incorporating hydrophobic sugar substituents exhibit efficacy against DAla-DLac-containing vancomycin-resistant enterococci, albeit by an undetermined mechanism. Contrasting models that invoke either cooperative dimerization and membrane anchoring or direct inhibition of bacterial transglycosylases have been proposed to explain the bioactivity of these glycopeptides. RESULTS: Affinity chromatography has revealed direct interactions between a semisynthetic hydrophobic vancomycin (DCB-PV), and select Escherichia coli membrane proteins, including at least six enzymes involved in peptidoglycan assembly. The N(4) vancosamine substituent is critical for protein binding. DCB-PV inhibits transglycosylation in permeabilized E. coli, consistent with the observed binding of the PBP-1B transglycosylase-transpeptidase. CONCLUSIONS: Hydrophobic vancomycins interact directly with a select subset of bacterial membrane proteins, suggesting the existence of discrete protein targets. Transglycosylase inhibition may play a role in the enhanced bioactivity of semisynthetic glycopeptides. PMID- 11731301 TI - Engineering inhibitors highly selective for the S1 sites of Ser190 trypsin-like serine protease drug targets. AB - BACKGROUND: Involved or implicated in a wide spectrum of diseases, trypsin-like serine proteases comprise well studied drug targets and anti-targets that can be subdivided into two major classes. In one class there is a serine at position 190 at the S1 site, as in urokinase type plasminogen activator (urokinase or uPA) and factor VIIa, and in the other there is an alanine at 190, as in tissue type plasminogen activator (tPA) and factor Xa. A hydrogen bond unique to Ser190 protease-arylamidine complexes between O gamma(Ser190) and the inhibitor amidine confers an intrinsic preference for such inhibitors toward Ser190 proteases over Ala190 counterparts. RESULTS: Based on the structural differences between the S1 sites of Ser190 and Ala190 protease-arylamidine complexes, we amplified the selectivity of amidine inhibitors toward uPA and against tPA, by factors as high as 220-fold, by incorporating a halo group ortho to the amidine of a lead inhibitor scaffold. Comparison of K(i) values of such halo-substituted and parent inhibitors toward a panel of Ser190 and Ala190 proteases demonstrates pronounced selectivity of the halo analogs for Ser190 proteases over Ala190 counterparts. Crystal structures of Ser190 proteases, uPA and trypsin, and of an Ala190 counterpart, thrombin, bound by a set of ortho (halo, amidino) aryl inhibitors and of non-halo parents reveal the structural basis of the exquisite selectivity and validate the design principle. CONCLUSIONS: Remarkable selectivity enhancements of exceptionally small inhibitors are achieved toward the uPA target over the highly similar tPA anti-target through a single atom substitution on an otherwise relatively non-selective scaffold. Overall selectivities for uPA over tPA as high as 980-fold at physiological pH were realized. The increase in selectivity results from the displacement of a single bound water molecule common to the S1 site of both the uPA target and the tPA anti-target because of the ensuing deficit in hydrogen bonding of the arylamidine inhibitor when bound in the Ala190 protease anti-target. PMID- 11731302 TI - Soft tissue deformation. PMID- 11731303 TI - Measurements and modelling of the compliance of human and porcine organs. AB - Stress-strain data obtained from animal and human tissue have several applications including medical diagnosis, assisting in surgical instrument design and the production of realistic computer-based simulators for training in minimal access surgery. Such data may also be useful for corroborating mathematical models of tissue response. This paper presents data obtained from ex-vivo and in vivo tissue indentation tests using a small indentor that is similar to instruments used in minimal access surgery. In addition, uniform stress tests provide basic material property data, via an exponential stress-strain law, to allow a finite element method to be used to predict the response for the non uniform stresses produced by the small indentor. Data are obtained from harvested pig liver and spleen using a static compliance probe. Data for human liver are obtained from volunteer patients, undergoing minor open surgery, using a sterile hand-held compliance probe. All the results demonstrate highly non-linear stress strain behaviour. Pig spleen is shown to be much more compliant than pig liver with mean elastic moduli of 0.11 and 4.0 MPa respectively. The right lobe of human liver had a mean elastic modulus of about 0.27 MPa. However, a single case of a diseased liver had a mean modulus of 0.74 MPa--nearly three times the stiffness. It was found that an exponential stress-strain law could accurately fit uniform stress test data and that subsequent finite element modelling for non uniform stress around a small indentor matched measured force characteristics. PMID- 11731305 TI - In vivo finite element model-based image analysis of pacemaker lead mechanics. AB - BACKGROUND: Fractures of implanted pacemaker leads are currently identified by inspecting radiographic images without making full use of a priori known material and structural information. Moreover, lead designers are unable to incorporate clinical image data into analyses of lead mechanics. METHODS: A novel finite element/active contour method was developed to quantify the in vivo mechanics of implanted leads by estimating the distributions of stress, strain, and traction using biplane videoradiographic images. The nonlinear equilibrium equations governing a thin elastic beam undergoing 3-D large rotation were solved using one dimensional isoparametric finite elements. External forces based on local image greyscale values were computed from each pair of images using a perspective transformation governing the relationship between the image planes. RESULTS: Cantilever beam forward solution results were within 0.2% of the analytic solution for a wide range of applied loads. The finite element/active contour model was able to reproduce the principal curvatures of a synthetic helix within 3% of the analytic solution and estimates of the helix's geometric torsion were within 20% of the analytic solution. Applying the method to biplane videoradiographic images of a lead acutely implanted in an anesthetized dog resulted in expected variations in curvature and bending stress between compliant and rigid segments of the lead. CONCLUSIONS: By incorporating knowledge about lead geometric and material properties, the 3-D finite element/active contour method regularizes the image reconstruction problem and allows for more quantitative and automatic assessment of implanted lead mechanics. PMID- 11731304 TI - Magnetic resonance elastography: non-invasive mapping of tissue elasticity. AB - Magnetic resonance elastography (MRE) is a phase-contrast-based MRI imaging technique that can directly visualize and quantitatively measure propagating acoustic strain waves in tissue-like materials subjected to harmonic mechanical excitation. The data acquired allows the calculation of local quantitative values of shear modulus and the generation of images that depict tissue elasticity or stiffness. This is significant because palpation, a physical examination that assesses the stiffness of tissue, can be an effective method of detecting tumors, but is restricted to parts of the body that are accessible to the physician's hand. MRE shows promise as a potential technique for 'palpation by imaging', with possible applications in tumor detection (particularly in breast, liver, kidney and prostate), characterization of disease, and assessment of rehabilitation (particularly in muscle). We describe MRE in the context of other recent techniques for imaging elasticity, discuss the processing algorithms for elasticity reconstruction and the issues and assumptions they involve, and present recent ex vivo and in vivo results. PMID- 11731306 TI - Talairach-Tournoux brain atlas registration using a metalforming principle-based finite element method. AB - In this paper, a novel non-rigid registration method is proposed for registration of the Talairach-Tournoux brain atlas with MRI images and the Schaltenbrand Wahren brain atlas. A metalforming principle-based finite element method with the large deformation problem is used to find the local deformation, in which finite element equations are governed by constraints in the form of displacements derived from the correspondence relationship between extracted feature points. Some detectable substructures, such as the cortical surface, ventricles and corpus callosum, are first extracted from MRI, forming feature points which are classified into different groups. The softassign method is used to establish the correspondence relationship between feature points within each group and to obtain the global transformation concurrently. The displacement constraints are then derived from the correspondence relationship. A metalforming principle-based finite element method with the large deformation problem is used in which finite element equations are reorganized and simplified by integrating the displacement constraints into the system equations. Our method not only matches the model to the data efficiently, but also decreases the degrees of freedom of the system and consequently reduces the computational cost. The method is illustrated by matching the Talairach-Tournoux brain atlas to MRI normal and pathological data and to the Schaltenbrand-Wahren brain atlas. We compare the results quantitatively between the force assignment-based method and the proposed method. The results show that the proposed method yields more accurate results in a fraction of the time taken by the previous method. PMID- 11731307 TI - Fast extraction of minimal paths in 3D images and applications to virtual endoscopy. AB - The aim of this article is to build trajectories for virtual endoscopy inside 3D medical images, using the most automatic way. Usually the construction of this trajectory is left to the clinician who must define some points on the path manually using three orthogonal views. But for a complex structure such as the colon, those views give little information on the shape of the object of interest. The path construction in 3D images becomes a very tedious task and precise a priori knowledge of the structure is needed to determine a suitable trajectory. We propose a more automatic path tracking method to overcome those drawbacks: we are able to build a path, given only one or two end points and the 3D image as inputs. This work is based on previous work by Cohen and Kimmel [Int. J. Comp. Vis. 24 (1) (1997) 57] for extracting paths in 2D images using Fast Marching algorithm. Our original contribution is twofold. On the first hand, we present a general technical contribution which extends minimal paths to 3D images and gives new improvements of the approach that are relevant in 2D as well as in 3D to extract linear structures in images. It includes techniques to make the path extraction scheme faster and easier, by reducing the user interaction. We also develop a new method to extract a centered path in tubular structures. Synthetic and real medical images are used to illustrate each contribution. On the other hand, we show that our method can be efficiently applied to the problem of finding a centered path in tubular anatomical structures with minimum interactivity, and that this path can be used for virtual endoscopy. Results are shown in various anatomical regions (colon, brain vessels, arteries) with different 3D imaging protocols (CT, MR). PMID- 11731308 TI - Modeling the 3D coronary tree for labeling purposes. AB - Coronary artery diseases are usually revealed using X-ray angiographies. Such images are complex to analyze because they provide a 2D projection of a 3D object. Medical diagnosis suffers from inter- and intra-clinician variability. Therefore, reliable software for the 3D reconstruction and labeling of the coronary tree is strongly desired. It requires the matching of the vessels in the different available angiograms, and an approach which identifies the arteries by their anatomical names is a way to solve this difficult problem. This paper focuses on the automatic labeling of the left coronary tree in X-ray angiography. Our approach is based on a 3D topological model, built from the 3D anthropomorphic phantom, Coronix. The phantom is projected under different angles of view to provide a data base of 2D topological models. On the other hand, the vessel skeleton is extracted from the patient's angiogram. The algorithm compares the skeleton with the 2D topological model which has the most similar vascular net shape. The method performs in a hierarchical manner, first labeling the main artery, then the sub-branches. It handles inter-individual anatomical variations, segmentation errors and image ambiguities. We tested the method on standard angiograms of Coronix and on clinical examinations of nine patients. We demonstrated successful scores of 90% correct labeling for the main arteries and 60% for the sub-branches. The method appears to be particularly efficient for the arteries in focus. It is therefore a very promising tool for the automatic 3D reconstruction of the coronary tree from monoplane temporal angiographic clinical sequences. PMID- 11731309 TI - A computational model of postoperative knee kinematics. AB - A mathematical model for studying the passive kinematics of total knee prostheses can be useful in computer-aided planning and guidance of total joint replacement. If the insertion location and neutral length of knee ligaments is known, the passive kinematics of the knee can be calculated by minimizing the strain energy stored in the ligaments at any angular configuration of the knee. Insertions may be found intraoperatively, or may come from preoperative 3D medical images. The model considered here takes into consideration the geometry of the prosthesis and patient-specific information. This model can be used to study the kinematics of the knee joint of a patient after total joint replacement. The model may be useful in preoperative planning, computer-aided intraoperative guidance, and the design of new prosthetic joints. PMID- 11731312 TI - Morphogenesis of Escherichia coli. AB - Morphogenesis of the rod-shaped Escherichia coli is determined by controlled growth of an exoskeleton made of murein (peptidoglycan). Recent insights in the growth strategy of the stress-bearing murein sacculus has contributed to our understanding of how the required concerted action of murein polymerizing and hydrolyzing enzymes is achieved. The proteins involved are coordinated by the formation of multienzyme complexes. In this review, we summarize the recent results on murein structure and metabolism. On the basis of these findings, we present a model that explains maintenance of the specific rod shape of E. coli. PMID- 11731313 TI - Bacterial ancestry of actin and tubulin. AB - The structural and functional resemblance between the bacterial cell-division protein FtsZ and eukaryotic tubulin was the first indication that the eukaryotic cytoskeleton may have a prokaryotic origin. The bacterial ancestry is made even more obvious by the findings that the bacterial cell-shape-determining proteins Mreb and Mbl form large spirals inside non-spherical cells, and that MreB polymerises in vitro into protofilaments very similar to actin. Recent advances in research on two proteins involved in prokaryotic cytokinesis and cell shape determination that have similar properties to the key components of the eukaryotic cytoskeleton are discussed. PMID- 11731314 TI - Organelle fission in eukaryotes. AB - The cellular machineries that power chloroplast and mitochondrial division in eukaryotes carry out the topologically challenging job of constricting and severing these double-membraned organelles. Consistent with their endosymbiotic origins, mitochondria in protists and chloroplasts in photosynthetic eukaryotes have evolved organelle-targeted forms of FtsZ, the prokaryotic ancestor of tubulin, as key components of their fission complexes. In fungi, animals and plants, mitochondria no longer utilize FtsZ for division, but several mitochondrial division proteins that localize to the outer membrane and intermembrane space, including two related to the filament-forming dynamins, have been identified in yeast and animals. Although the reactions that mediate organelle division are not yet understood, recent progress in uncovering the constituents of the organelle division machineries promises rapid advancement in our understanding of the biochemical mechanisms underlying the distinct but related processes of chloroplast and mitochondrial division in eukaryotes. PMID- 11731315 TI - Spatial regulation of cytokinesis in bacteria. AB - Cytokinesis in bacteria such as Escherichia coli is orchestrated by FtsZ, a tubulin-like protein that forms a circumferential Z ring at the division site. The Z ring then recruits a number of other essential cell division proteins, ultimately assembling the cytokinetic machine that splits the cell. It has been known for some time that the MinCDE proteins and the bacterial nucleoid provide positional information to negatively regulate cytokinesis. Recently, direct visualization of Z rings and Min proteins in whole cells have contributed important new insights into the molecular mechanisms behind this fundamental cellular process. This review summarizes and integrates these insights. PMID- 11731316 TI - Chromosome segregation. AB - The ability to visualise specific genes and proteins within bacterial cells is revolutionising knowledge of chromosome segregation. The essential elements appear to be the driving force behind DNA replication, which occurs at fixed cellular positions, the condensation of newly replicated DNA by a chromosome condensation machine located at the cell 1/4 and 3/4 positions, and molecular machines that act at midcell to allow chromosome separation after replication and movement of the sister chromosomes away from the division septum prior to cell division. This review attempts to provide a perspective on current views of the bacterial chromosome segregation mechanism and how it relates to other cellular processes. PMID- 11731317 TI - Septation and chromosome segregation during sporulation in Bacillus subtilis. AB - The early stages of sporulation in Bacillus subtilis incorporate a modified, highly asymmetric cell division. It is now clear that most, if not all, of the components of the vegetative division machinery are used also for asymmetric division. However, the machinery for chromosome segregation may differ significantly between vegetative growth and sporulation. Several interesting checkpoint mechanisms couple cell cycle events to gene expression early in sporulation. This review summarises important advances in the understanding of chromosome segregation and cell division at the onset of sporulation in B.subtilis in the past three years. PMID- 11731318 TI - Regulation of sporulation in Streptomyces coelicolor A3(2): a checkpoint multiplex? AB - By complementing developmental mutants of Streptomyces coelicolor A3(2), at least 15 regulatory genes for sporulation have been identified and studied at the molecular level, and some of their intracellular interactions have been characterised. Extensive interplay of the regulatory cascade with metabolic, morphological, homeostatic and stress-related checkpoints is emerging. PMID- 11731319 TI - Cell cycle control of cell morphogenesis in Caulobacter. AB - In Caulobacter crescentus, morphogenic events, such as cytokinesis, the establishment of asymmetry and the biogenesis of polar structures, are precisely regulated during the cell cycle by internal cues, such as cell division and the initiation of DNA replication. Recent studies have revealed that the converse is also true. That is, differentiation events impose regulatory controls on other differentiation events, as well as on progression of the cell cycle. Thus, there are pathways that sense the assembly of structures or the localization of complexes and then transduce this information to subsequent biogenesis or cell cycle events. In this review, we examine the interplay between flagellar assembly and the C. crescentus cell cycle. PMID- 11731320 TI - The septin cortex at the yeast mother-bud neck. AB - A specialized cortical domain is organized by the septins at the necks of budding yeast cells. Recent findings suggest that this domain serves as a diffusion barrier and also as a local cell-shape sensor. We review these findings along with what is known about the organization of the septin cortex and its regulation during the cell cycle. PMID- 11731321 TI - Assembly and regulation of the cytokinetic apparatus in budding yeast. AB - The identification of an actomyosin-based contractile ring in budding yeast has recently established this organism as a general model for studying cytokinesis. Work over the past three years has provided important new insights into the conserved mechanisms underlying the assembly and regulation of the cytokinetic structures. This review covers the recent progress in studying cytokinesis in budding yeast. PMID- 11731322 TI - The social life of actin and microtubules: interaction versus cooperation. AB - The actin and microtubule cytoskeletons play key roles in cell polarity, spindle orientation and nuclear movement. Recent work in fungal systems has identified potential "functional links" between these cytoskeletal systems. This review discusses molecular mechanisms through which these links may be established. PMID- 11731323 TI - Modular complexes that regulate actin assembly in budding yeast. AB - The actin cytoskeleton of budding yeast contains an extensive set of actin associated proteins with conserved mammalian counterparts. For more than 20 years, yeast has been used as a model organism to dissect the in vivo functions of these factors, revealing an intricate web of genetic interactions in the cell. Now, a surge of biochemical reports is defining the physical interactions and activities of these proteins and providing mechanistic insights into their cellular roles. The emerging view is that most actin-associated proteins do not act alone but, rather, associate to form modular protein complexes that regulate actin assembly and organization. PMID- 11731324 TI - Cytokinesis and the contractile ring in fission yeast. AB - The fission yeast Schizosaccharomyces pombe provides a genetic model system for the study of cytokinesis. As in many eukaryotes, cell division in the fission yeast requires an actin-myosin-based contractile ring. Numerous components of the contractile ring that function in ring assembly, positioning and contraction have been characterized. Many of these proteins are evolutionarily conserved, suggesting that common molecular mechanisms may govern aspects of eukaryotic cell division. Recent advances in the assembly and placement of the contractile ring are discussed. In particular, major findings have been made in the characterization of myosins in cytokinesis, and in how the cell division site may be positioned by the nucleus. PMID- 11731325 TI - Cell cycle control of yeast filamentous growth. AB - Great progress has been made toward dissecting the signal transduction pathways and transcriptional outputs regulating yeast pseudohyphal growth. However, the mechanism underlying polarized morphogenesis in filamentous growth remains unclear. A synthesis of the data suggests that the ultimate target of these pathways is to repress the activity of the mitotic cyclin Clb2 as an antagonist of polarized growth. Here, we discuss how this cell cycle regulation, in concert with control of transcription, ubiquitin-dependent proteolysis and cytoskeletal polarity, may mediate the switch to filamentous growth. PMID- 11731327 TI - Septum formation in Aspergillus nidulans. AB - Filamentous fungi form multicellular hyphae that are partitioned by septa. In A. nidulans, septum formation requires the assembly of a septal band following the completion of mitosis. Recent observations show that this band is a dynamic structure composed of actin, a septin and a formin. In addition, assembly is dependent upon a conserved protein kinase cascade that regulates mitotic exit and septation in yeast. Hyphal differentiation may reflect the regulation of this cascade by cyclin-dependent kinase activity. In this review, the dynamics and regulation underlying the assembly of the septal band are discussed. PMID- 11731326 TI - Transcriptional control of dimorphism in Candida albicans. AB - Candida albicans uses a network of multiple signaling pathways to control the yeast-->hypha transition. These include a mitogen-activated protein kinase pathway through Cph1, the cAMP-dependent protein kinase pathway via Efg1, a pH responsive pathway through Rim101, the Tup1-mediated repression through Rfg1 and Nrg1, and pathways represented by transcription factors Cph2, Tec1 and Czf1. These pathways control the transcription of a common set of hypha-specific genes, many of which encode known virulence factors. The link between the signaling pathways and hyphal elongation is currently unknown, but there is evidence to suggest that Cdc42 likely plays a key role in hyphal morphogenesis. Unlike pseudohyphal growth in Saccharomyces cerevisiae, hyphal elongation is regulated independently of the cell cycle. Cellular differences between pseudohyphae and hyphae are further revealed by septin localization. PMID- 11731328 TI - The effects of dietary conjugated nonadecadienoic acid on body composition in mice. AB - A 19-carbon conjugated diene, conjugated nonadecadienoic acid (CNA), inhibited heparin-releasable lipoprotein lipase and reduced lipid stores in 3T3-L1 adipocytes similarly to conjugated linoleic acid (CLA). When fed to growing mice (0.3% of diet) CNA reduced body fat by 81% whereas CLA reduced body fat by 25%. CLA and CNA differ in length by one carbon atom so they are unlikely to share a common metabolite to account for these observations. PMID- 11731329 TI - Cellular cholesterol efflux. AB - Efflux of free cholesterol (FC) continues even when cellular FC mass is unchanged. This reflects a recirculation of preformed FC between cells and extracellular fluids which has multiple functions in cell biology including receptor recycling and signaling as well as cellular FC homeostasis. Total FC efflux is heterogeneous. Simple diffusion to mature high density lipoprotein (HDL), mainly via albumin as intermediate, initiates FC net transport driven by plasma lecithin:cholesterol acyltransferase activity. A second major efflux component reflects protein-facilitated transport from cell surface domains (caveolae, rafts) driven by FC binding to lipid-poor, pre-beta-migrating HDL (pre beta-HDL). Facilitated efflux from caveolae, unlike simple diffusion, is highly regulated. Neither ABC1 (the protein defective in Tangier disease) nor other ATP dependent transporters now appear likely to contribute directly to FC efflux. Their role is limited to the initial formation of a particle precursor to circulating pre-beta-HDL, which recycles without further lipid input from ATP dependent transporter proteins. Lipid-free apolipoprotein A-I, previously considered a surrogate for pre-beta-HDL, has a reactivity much lower than that of native lipoprotein FC acceptors. PMID- 11731330 TI - Phosphoinositides, key molecules for regulation of actin cytoskeletal organization and membrane traffic from the plasma membrane. AB - Phosphoinositide plays a critical role not only in generating second messengers, such as inositol 1,4,5-trisphosphate and diacylglycerol, but also in modulating a variety of cellular functions including cytoskeletal organization and membrane trafficking. Many inositol lipid kinases and phosphatases appear to regulate the concentration of a variety of phosphoinositides in a specific area, thereby inducing spatial and temporal changes in their availability. For example, local concentration changes in phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) in response to extracellular stimuli cause the reorganization of actin filaments and a change in cell shape. PI(4,5)P(2) uncaps the barbed end of actin filaments and increases actin nucleation by modulating a variety of actin regulatory proteins, leading to de novo actin polymerization. PI(4,5)P(2) also plays a key role in membrane trafficking processes. In endocytosis, PI(4,5)P(2) targets clathrin associated proteins to endocytic vesicles, leading to clathrin-coated pit formation. On the contrary, PI(4,5)P(2) must be dephosphorylated when they shed clathrin coats to fuse endosome. Thus, through regulating actin cytoskeleton organization and membrane trafficking, phosphoinositides play crucial roles in a variety of cell functions such as growth, polarity, movement, and pattern formation. PMID- 11731331 TI - gamma-Linolenic acid and eicosapentaenoic acid induce modifications in mitochondrial metabolism, reactive oxygen species generation, lipid peroxidation and apoptosis in Walker 256 rat carcinosarcoma cells. AB - The polyunsaturated fatty acids gamma-linolenic acid (GLA) and eicosapentaenoic acid (EPA) are cytotoxic to tumour cells. GLA inhibits Walker 256 tumour growth in vivo, causing alterations in mitochondrial ultrastructure and cellular metabolism. The objective of the present study was to investigate the mechanisms behind fatty acid inhibition of Walker 256 tumour growth under controlled in vitro conditions. At a concentration of 150 microM, both GLA and EPA caused a decrease in cell proliferation and an increase in apoptotic index. Increases in reactive oxygen species (ROS) and lipid peroxide production were identified, as well as alterations in energy metabolism and the deposition of large amounts of triacylglycerol in the form of lipid droplets. Mitochondrial respiratory chain complexes I+III and IV had significantly decreased activity and mitochondrial membrane potential was greatly diminished. Intracellular ATP concentrations were maintained at 70-80% of control values despite the decreased mitochondrial function, which may be in part due to increased utilisation of glucose for ATP generation. Cytochrome c release from mitochondria was found, as was caspase-3 like activation. DNA fragmentation in situ revealed many apoptotic events within the cell population. The mechanism(s) by which ROS and lipid peroxides induce apoptosis remains unclear, but the effects of GLA and EPA appear to involve the mitochondrial pathway of apoptosis induction leading to cytochrome c release, caspase activation, loss of mitochondrial membrane potential and DNA fragmentation. PMID- 11731332 TI - Regulation of lipid metabolism and gene expression by fenofibrate in hamsters. AB - Fenofibrate is a potent hypolipidemic agent that lowers plasma lipid levels and may thus decrease the incidence of atherosclerosis. Here we investigated the molecular mechanism of fenofibrate's hypolipidemic action by characterizing its in vivo effects on the expression of mRNAs and the activities of pivotal enzymes in cholesterol and triglyceride metabolism in the hamster. Treatment of hamsters with fenofibrate led to a dose-dependent reduction in serum cholesterol concentrations. Studies on the incorporation of [(14)C]acetate and [(14)C]mevalonate into cholesterol suggested that this effect occurs primarily through inhibition of cholesterol biosynthesis at steps prior to mevalonate. Fenofibrate decreased levels of hepatic enzyme activities and mRNAs for 3-hydroxy 3-methylglutaryl coenzyme A (HMG CoA) synthase and HMG CoA reductase. A potential mechanism for transcriptional regulation of these enzymes is via SREBP-2 that we found to be suppressed 2-fold by fenofibrate. Fenofibrate also lowered circulatory triglyceride levels. In keeping with the effect, we observed strong suppression of fatty acid synthase, acetyl-CoA carboxylase and apolipoprotein C III mRNA and stimulation of lipoprotein lipase and acyl-CoA oxidase mRNA in the liver of fenofibrate-treated hamsters. These observations suggest that the effect of fenofibrate on triglyceride metabolism is likely to be a result of both decreased fatty acid synthesis and increased lipoprotein lipase and acyl-CoA oxidase gene expression in the liver. Surprisingly, alterations in lipoprotein lipase, acyl-CoA oxidase, acetyl-CoA carboxylase, and apolipoprotein C-III could not be observed in hamster hepatocytes incubated with fenofibric acid in vitro. These observations raise the possibility that changes in these genes may be secondary to the metabolic alterations occurring in animals but not in cultured cells and thus that the effect of fenofibrate on these genes may be indirect. PMID- 11731333 TI - Influence of conjugated linoleic acid on body composition and target gene expression in peroxisome proliferator-activated receptor alpha-null mice. AB - The mechanisms underlying the beneficial effects of conjugated linoleic acid (CLA) are unknown, but one hypothesis is that they are mediated by the nuclear receptor, peroxisome proliferator-activated receptor (PPARalpha). In this work, the effect of dietary CLA on body weight gain, body composition, serum lipids and tissue specific PPAR target gene expression was examined in PPARalpha-null mice. Male wild-type or PPARalpha-null mice were fed either a control diet or one containing 0.5% CLA for a period of 4 weeks. Weight gain in wild-type and PPARalpha-null mice fed CLA was similar, and significantly less than controls. Whole body fat content was lower in wild-type and PPARalpha-null mice while whole body protein content was increased in both genotypes fed CLA compared to controls. Serum triglycerides were lowered in both genotypes as a result of dietary CLA. While CLA feeding resulted in specific activation of PPARalpha in liver, alterations in liver, adipose and muscle mRNAs were also found that were independent of PPARalpha genotype including those encoding uncoupling proteins (UCPs), mitochondrial fatty acid oxidizing enzymes, and fatty acid transporter. These results demonstrate that despite specific activation of PPARalpha-dependent gene expression, the influence of CLA on body composition appears to be independent of PPARalpha. Further, CLA causes increased levels of mRNAs encoding lipid metabolizing and mitochondrial uncoupling proteins that likely contribute to the mechanisms underlying reduced fat/increased lean body mass resulting from consumption of dietary CLA. PMID- 11731334 TI - Autologous lymphocyte-monocyte co-culture increases NMR-visible and cytoplasmic lipids in the absence of increased markers of lymphocyte activation. AB - Alterations in nuclear magnetic resonance (NMR)-visible lipid, morphometric lipid volume fraction, distribution of subcellular lipid droplets and activation antigen expression were examined in human peripheral blood lymphocytes, activated using phorbol myristate acetate (PMA) and ionomycin or by co-culture with autologous monocytes. PMA/Ionomycin treatment caused significant time-dependent increases in mobile lipid and in oil red O-positive lipid droplets that were accompanied by lymphocyte proliferation and increases in activation antigens, CD25, CD69 and CD71. Co-culture of lymphocytes and monocytes also induced significant increases in NMR-visible lipid signals and cytoplasmic lipid droplets, but in contrast, no correspondent increases in activation antigens were observed. Strong correlations were observed between the intensity of the NMR signal and the percentage of total cells containing lipid droplets (r=0.95) and the morphometric lymphocyte lipid volume fraction (r=0.80), indicating that the droplets were the source of the mobile lipid signal. Lipid droplets in PMA/Ionomycin-treated cells were evenly distributed throughout the population, but in co-cultures, only lymphocytes in close proximity to monocytes with lipid droplets contained oil red O-positive lipid. This data shows that the NMR-visible mobile lipid signal observed in lymphocytes co-cultured with monocytes is not directly dependent on either proliferation or the upregulation of activation antigens, similar to the previously observed response of T cells exposed to antibodies to the T cell receptor. PMID- 11731336 TI - Enzymatic and non-enzymatic lipid peroxidation in leaf development. AB - Enzymatic and non-enzymatic lipid peroxidation has been implicated in programmed cell death, which is a major process of leaf senescence. To test this hypothesis we developed a high-performance liquid chromatography (HPLC) method for a simultaneous analysis of the major hydro(pero)xy polyenoic fatty acids. Quantities of lipid peroxidation products in leaves of different stages of development including natural senescence indicated a strong increase in the level of oxygenated polyenoic fatty acids (PUFAs) during the late stages of leaf senescence. Comprehensive structural elucidation of the oxygenation products by means of HPLC, gas chromatography/mass spectrometry and (1)H nuclear magnetic resonance suggested a non-enzymatic origin. However, in some cases a small share of specifically oxidized PUFAs was identified suggesting involvement of lipid peroxidizing enzymes. To inspect the possible role of enzymatic lipid peroxidation in leaf senescence, we analyzed the abundance of lipoxygenases (LOXs) in rosette leaves of Arabidopsis. LOXs and their product (9Z,11E,13S,15Z) 13-hydroperoxy-9,11,15-octadecatrienoic acid were exclusively detected in young green leaves. In contrast, in senescing leaves the specific LOX products were overlaid by large amounts of stereo-random lipid peroxidation products originating from non-enzymatic oxidation. These data indicate a limited contribution of LOXs to total lipid peroxidation, and a dominant role of non enzymatic lipid peroxidation in late stages of leaf development. PMID- 11731335 TI - Studies on the transport of acetyl groups from peroxisomes to mitochondria in isolated liver cells oxidizing the polyunsaturated fatty acid 22:4n-6. AB - The oxidation of the fatty acid [1-(14)C]22:4n-6 was studied in isolated hepatocytes. Labeled acetate was the main acid soluble product identified by HPLC after short incubation periods. At low substrate concentrations and longer incubations [(14)C]acetate was gradually replaced by labeled beta hydroxybutyrate, acetoacetate and oxaloacetate/malate. Preincubation with 2 tetradecylglycidic acid (TDGA), an inhibitor of mitochondrial fatty acid oxidation, did not reduce the oxidation but acetate was the only product recovered. TDGA also strongly inhibited the metabolism of added [1-(14)C]acetate to mitochondrial oxidation products. During the preparation procedure of hepatocytes the cellular L-carnitine concentration was decreased but it was restored after preincubation with L-carnitine. With low [1-(14)C]22:4n-6, concentrating a low level of [(14)C]acetate and high levels of labeled mitochondrial oxidation products were recovered after preincubation with L carnitine. A small amount of [(14)C]acetylcarnitine was also detected under this incubation condition. The results suggest that a significant part of labeled acetyl groups from the peroxisomal oxidation of [1-(14)C]22:4n-6 is transported to the mitochondria as free acetate. Moreover, the results also suggest that L carnitine at physiological concentrations may facilitate the transport of part of the acetyl groups from peroxisomes to mitochondria as acetylcarnitine. However, the possibility that an increased cellular L-carnitine concentration may stimulate oxidation of [1-(14)C]22:4n-6 in mitochondria could not be excluded. PMID- 11731337 TI - Molecular cloning and structural analysis of human sterol C5 desaturase. AB - Sterol C5 desaturase (SC5D) converts lathosterol to 7-dehydrocholesterol in cholesterol biosynthesis. In this study, we investigated the genome structure of SC5D and transcription of the human SC5D gene to try to elucidate the mechanism by which cholesterol synthesis is regulated. The SC5D gene had a structural gene from a single copy from genome DNA that contained five exons and four introns. The human SC5D was found to be located in chromosome 11q24.2-24.3 by fluorescence in situ hybridization mapping. Human SC5D mRNA transcripts, of which the major transcript was a 2 kb and the minor transcripts were 8 kb and 1.4 kb mRNA transcripts, were detected in almost all of the tissues examined. The human SC5D gene contained a GC box instead of a TATA box upstream of the transcript start sites. Human SC5D transcription started from several transcription start sites, and the first start site was located 31 bp upstream of the translation start site (ATG). The expression level of SC5D mRNA extracted from human liver carcinoma cells decreased as the amount of cholesterol added to the culture medium was increased. Inhibition of SC5D transcription was ascribed to the suppression of promoter activity of SC5D. PMID- 11731338 TI - Main factors governing the transfer of carotenoids from emulsion lipid droplets to micelles. AB - Carotenoids might lower the incidence of several diseases, yet the mechanisms governing their intestinal absorption are still poorly understood. The aim was to identify and study the main factors governing the transfer of carotenoids from emulsion lipid droplets to mixed micelles, presumed to be a key step in carotenoid absorption. An in vitro model was devised to measure the transfer, and a factorial design was applied to identify the main factors affecting the transfer. Experiments were then conducted to assess the effect of physiological variations of the main factors on the transfer efficiency. Finally, different carotenoids were simultaneously incorporated in emulsion lipid droplets to determine whether they interacted during the transfer. The factorial design gave three factors that significantly affected the transfer: type of carotenoid, pH, and bile lipid concentration. The transfer was (i) inversely related to carotenoid hydrophobicity, (ii) maximum between pH 6 and 7, (iii) maximum from 2 mmol/l bile salts, (iv) impaired by other carotenoids in the case of carotenes, but not in the case of xanthophylls. The transfer mainly depends on carotenoid hydrophobicity, pH, and bile lipid concentration. Physiological variations in pH and bile lipid concentration markedly affect the transfer. Both carotenes and xanthophylls can impair the transfer of carotenes, whereas they have apparently no effect on the transfer of xanthophylls. PMID- 11731339 TI - Binding properties of Echinococcus granulosus fatty acid binding protein. AB - EgFABP1 is a developmentally regulated intracellular fatty acid binding protein characterized in the larval stage of parasitic platyhelminth Echinococcus granulosus. It is structurally related to the heart group of fatty acid binding proteins (H-FABPs). Binding properties and ligand affinity of recombinant EgFABP1 were determined by fluorescence spectroscopy using cis- and trans-parinaric acid. Two binding sites for cis- and trans-parinaric acid were found (K(d(1)) 24+/-4 nM, K(d(2)) 510+/-60 nM for cis-parinaric acid and K(d(1)) 32+/-4 nM, K(d(2)) 364+/-75 nM for trans-parinaric). A putative third site for both fatty acids is discussed. Binding preferences were determined using displacement assays. Arachidonic and oleic acids presented the highest displacement percentages for EgFABP1. The Echinococcus FABP is the unique member of the H-FABP group able to bind two long chain fatty acid molecules with high affinity. Structure-function relationships and putative roles for EgFABP1 in E. granulosus metabolism are discussed. PMID- 11731340 TI - Desiccation, cryopreservation and water relations parameters of white spruce (Picea glauca) and interior spruce (Picea glauca x engelmannii complex) somatic embryos. AB - Effects of drying and cryopreservation on survival of spruce (Picea glauca (Moench) Voss and Picea glauca x engelmannii complex) somatic embryos (SEs) were investigated with the aim of developing simple and robust protocols for embryo storage. Somatic embryos dried over salt solutions of known water potential (Psi) survived removal of virtually all free water, to a relative water content (RWC) of approximately 0.13, a value similar to that for spruce zygotic embryos from dry seed. Desiccated SEs also survived subsequent freezing in liquid nitrogen, without the addition of cryoprotectant or pre-culture steps. Highest survival (> 80%) after freezing in liquid nitrogen was in embryos pre-dried to Psi of -15 to 20 MPa, which yielded RWC close to predicted bound (apoplastic) water values. Low (< 35%) or no survival after freezing was observed in embryos pretreated at higher Psi (above -5 MPa) or at very low Psi (-540 MPa, using silica gel), respectively. PMID- 11731341 TI - Leaf gas exchange and carbohydrates in tropical trees differing in successional status in two light environments in central Amazonia. AB - Monoculture and mixed stands of mahogany (Swietenia macrophylla King) and tonka bean (Dipteryx odorata Willd.) trees were established on degraded land in central Amazonia to compare leaf gas exchange parameters between plants grown in sunlight in an open field and in shade beneath a balsa wood (Ochroma pyramidale Cav.) canopy. Shading increased specific leaf area by about 50% in both species. Shading reduced height and diameter growth of S. macrophylla, whereas it increased these parameters for D. odorata. Light-saturated photosynthesis (Amax), stomatal conductance (gs) and transpiration (E) were higher in S. macrophylla than in D. odorata. In S. macrophylla, Amax was higher in sun leaves (12.9 +/- 0.9 micromol m-2 s-1) than in shade leaves (10.2 +/- 1.0 micromol m-2 s-1), whereas light environment had no significant effect on Amax of D. odorata. In both species, CO2-saturated photosynthesis (Apot) was higher in sun leaves (22 +/ 1.4 micromol m-2 s-1) than in shade leaves (17-20 +/- 0.8 micromol m-2 s-1). Stomatal conductance and E increased in sun leaves of S. macrophylla by 45 and 38%, respectively, whereas light environment did not affect gs and E of D. odorata. Photorespiration rates (Pr) varied from 28 to 36% of net photosynthesis (A) at ambient atmospheric CO2 concentration (Ca) but declined to around 7% of A at higher Ca. Leaf dark respiration (Rd) and the CO2 compensation point (Gamma) were lower in shade-grown plants than in open-grown plants. Compared with shade grown plants, total nonstructural carbohydrate (TNC) concentrations increased by 56% in S. macrophylla and by 30% in D. odorata in the open field. Leaf nitrogen (NL) concentrations were higher in open-grown plants than in shade-grown plants of both species, and TNC and NL concentrations were negatively correlated (r = 0.77). High TNC accumulation in S. macrophylla in the open field suggests that photosynthesis is unlikely to be growth-limiting at this site. We conclude that photosynthetic parameters of D. odorata are less affected by light environment than those of S. macrophylla, indicating that D. odorata has lower physiological plasticity to light than S. macrophylla. PMID- 11731342 TI - Growth of conifer seedlings on organic and inorganic nitrogen sources. AB - Effects of organic and inorganic nitrogen sources on growth and mineral nutrient concentrations of Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings were compared in a 100-day experiment in a greenhouse. Seedlings were grown in pots containing peat. Nutrient solutions differing in ammonium, nitrate, arginine and glycine composition were supplied to the seedlings at three nitrogen (N) concentrations: 1, 3 and 10 mM. We used dual (13C, 15N) and single (15N) isotopic labeling to determine the uptake of organic and inorganic N at the end of the experiment. Seedling dry weights and mineral nutrient concentrations of the needles showed that both conifer species were able to grow well and maintain nutrient balance on all investigated N forms except for the ammonium-dominated nutrient mixtures at the 10 mM N concentration. In Scots pine, no significant differences in dry weights were found between seedlings grown on the amino acids and seedlings grown on a commercial fertilizer containing 61.5% NO(3-)-N and 38.5% NH(4+)-N. Isotopic labeling of seedlings indicated that uptake rates of arginine-N, glycine-N and NH(4+)-N were similar, and 7-8 times greater than uptake rates for NO(3-)-N in both species. In Scots pine seedlings, 100% of arginine-N, and at least 67% of glycine-N was derived from the uptake of intact amino acids through seedling roots or mycorrhizae. Corresponding figures for Norway spruce were 83% for arginine and 96% for glycine. The gas chromatography-mass spectrometry analyses confirmed the presence of intact labeled molecules of both arginine and glycine in seedlings. We conclude that arginine and glycine are comparable to inorganic N as N sources for growth of conifer seedlings. PMID- 11731343 TI - Carbon uptake, growth and resource-use efficiency in one invasive and six native Hawaiian dry forest tree species. AB - Photosynthetic gas exchange, nitrogen- and water-use efficiency, leaf water potential and seasonal patterns of leaf production were studied in seven, dominant dry-forest species from the island of Lana'i, Hawaii, including the rapidly colonizing, non-native Schinus terebinthifolius (Raddi). We evaluated whether unique physiological characteristics of the invasive species explain its capacity to rapidly invade dry forests throughout the Hawaiian Islands. Apparent anomalies in stable carbon isotope data (delta13C) relative to other results led us to study effects of environmental conditions and physiological performance during leaf expansion on delta13C. Species that expanded all their foliage at the beginning of the wet season had more negative leaf delta13C values during the dry season than species with continuous leaf expansion. Among species, S. terebinthifolius had a strong seasonal pattern of leaf production and the most negative delta13C (-29 per thousand). With respect to almost every trait measured, S. terebinthifolius fell at an end of the range of values for the native species. Rapid growth of S. terebinthifolius in this ecosystem may be partially explained by its high maximum CO2 assimilation rates (15 micromol m-2 s 1), low leaf mass per area, high photosynthetic nitrogen-use efficiency per unit leaf mass or area and large decrease in stomatal conductance during the dry season. Relative to the native species, the invasive species exhibited striking phenotypic plasticity, including high rates of stem growth and water and CO2 uptake during the wet season, and maintenance of leaves and high leaf water potentials, as a result of reduced water loss, during the dry season, enabling it to utilize available resources effectively. PMID- 11731344 TI - Characterization of the responses of cork oak (Quercus suber) to iron deficiency. AB - We studied responses of cork oak (Quercus suber L.) to iron (Fe) deficiency by comparing seedlings grown hydroponically in nutrient solution with and without Fe. Seedlings grown without Fe developed some responses typical of the Strategy I group of Fe-efficient plants, including two- and fourfold increases in plasma membrane ferric chelate reductase activity of root tips after 2 and 4 weeks of culture in the absence of Fe, respectively. Moreover, seedlings grown hydroponically for 2 weeks without Fe caused marked decreases in the pH of the nutrient solution, indicating that root plasma membrane ATPase activity was induced by Fe deficiency. Iron deficiency also caused marked decreases in leaf chlorophyll and carotenoid concentrations, and chlorophyll concentrations were decreased more than carotenoid concentrations. Iron deficiency resulted in an 8% decrease in the dark-adapted efficiency of photosystem II and a 43% decrease in efficiency of photosystem II at steady-state photosynthesis. No major root morphological changes were observed in seedlings grown without Fe, although seedlings grown in Fe-deficient nutrient solution had light-colored roots in contrast to the dark brown color of control roots. PMID- 11731345 TI - Light-mediated constraints on leaf function correlate with leaf structure among deciduous and evergreen tree species. AB - Leaf structure has been shown to be an important determinant of leaf photosynthetic characteristics, yet the nature of this relationship remains ambiguous. It has been suggested that intra-leaf shading of chloroplasts may explain the negative influence of increasing leaf thickness/density on mass-based photosynthesis. To explore further the importance of light-mediated functional limitations conferred by leaf structure, we examined photosynthetic responses to high unidirectional and bidirectional irradiances among broad- and needle-leaves ranging widely in mass per area (LMA). Except for leaves with the lowest LMA (< 40 g m-2), photosynthesis increased in bidirectional irradiance, and the ratio of unidirectional to bidirectional photosynthesis (Au/Ab) was strongly and negatively related to LMA, approaching 0.5 for the thickest/densest leaves. Bidirectional illumination also increased stomatal conductance, but the magnitude of stomatal response was modest and uncorrelated with LMA. Consequently, the ratio of intercellular CO2 partial pressures during unidirectional versus bidirectional irradiation was positively related to LMA. Hence, it appears that many C3 leaves may not be "light saturated" under high unidirectional illumination. The negative, exponential Au/Ab-LMA relationship in this study supports the notion that, as LMA increases, an increasing fraction of leaf chloroplasts are functioning at subsaturating irradiance under unidirectional light. PMID- 11731346 TI - Mortality statistics in psychiatry. PMID- 11731347 TI - Adjustment disorders: fault line in the psychiatric glossary. PMID- 11731348 TI - Cycle of child sexual abuse: links between being a victim and becoming a perpetrator. AB - BACKGROUND: There is widespread belief in a 'cycle' of child sexual abuse, but little empirical evidence for this belief. AIMS: To identify perpetrators of such abuse who had been victims of paedophilia and/or incest, in order to: ascertain whether subjects who had been victims become perpetrators of such abuse; compare characteristics of those who had and had not been victims; and review psychodynamic ideas thought to underlie the behaviour of perpetrators. METHOD: Retrospective clinical case note review of 843 subjects attending a specialist forensic psychotherapy centre. RESULTS: Among 747 males the risk of being a perpetrator was positively correlated with reported sexual abuse victim experiences. The overall rate of having been a victim was 35% for perpetrators and 11% for non-perpetrators. Of the 96 females, 43% had been victims but only one was a perpetrator. A high percentage of male subjects abused in childhood by a female relative became perpetrators. Having been a victim was a strong predictor of becoming a perpetrator, as was an index of parental loss in childhood. CONCLUSIONS: The data support the notion of a victim-to-victimiser cycle in a minority of male perpetrators but not among the female victims studied. Sexual abuse by a female in childhood may be a risk factor for a cycle of abuse in males. PMID- 11731351 TI - Mental disorders and cause-specific mortality. AB - BACKGROUND: The impact of clinically diagnosed mental disorders on mortality in the general population has not been established. Aims To examine mental disorders for their prediction of cause-specific mortality. METHOD: Mental disorders were determined using the 36-item version of the General Health Questionnaire and the Present State Examination in a nationally representative sample of 8000 adult Finns. RESULTS: During the 17-year follow-up period 1597 deaths occurred. The presence of a mental disorder detected at baseline was associated with an elevated mortality rate. The relative risk in men was 1.6(95% confidence interval 1.3-1.8) and in women, 1.4 (95% Cl 1.2-1.6). In men and women with schizophrenia the relative risks of death during the follow-up period were 3.3 (95% Cl 2.3-4.9) and 2.3 (95% Cl 1.3-3.8) respectively, compared with the rest of the sample. In both men and women with schizophrenia the risk of dying of respiratory disease was increased, but the risk of dying of cardiovascular disease was increased only in men with neurotic depression. CONCLUSIONS: Schizophrenia and depression are associated with an elevated risk of natural and unnatural deaths. PMID- 11731352 TI - Extrastriatal and striatal D(2) dopamine receptor blockade with haloperidol or new antipsychotic drugs in patients with schizophrenia. AB - BACKGROUND: Both traditional and atypical antipsychotics have been hypothesised to be effective in schizophrenia through limbic and cortical D(2) dopamine receptor blockade. AIMS: To investigate this hypothesis with the D(2)/D(3) selective positron emission tomography (PET) probe [(76)Br]-FLB457. METHOD: PET scans were performed on 6 controls and 18 patients with schizophrenia treated with haloperidol or with risperidone, clozapine, amisulpride or olanzapine. RESULTS: The D(2) dopamine receptor blockade was high in the temporal cortex with both haloperidol and atypical antipsychotics. The atypicals, however, induced a significantly lower D(2) binding index than haloperidol in the thalamus and in the striatum. CONCLUSIONS: Results suggest that cortical D(2) dopamine receptors are a common target of traditional and atypical antipsychotics for therapeutic action. Higher in vivo binding to the D(2) receptors in the cortex than in the basal ganglia is suggested as an indicator of favourable profile for a putative antipsychotic compound. PMID- 11731353 TI - Comorbid non-alcohol substance misuse among people with schizophrenia: epidemiological study in central London. AB - BACKGROUND: Few epidemiological studies have assessed the extent and nature of comorbid non-alcohol substance misuse in people with schizophrenia in the community in the UK. AIMS: To study the extent and nature of comorbid non-alcohol substance misuse in people with schizophrenia in central London. METHOD: Subjects were identified in an epidemiological census survey of South Westminster. Standardised assessment of each subject included demographic data, ratings of mental state and movement disorder and questioning about drug and alcohol misuse. RESULTS: Individuals with schizophrenia or related psychoses were identified (n=352) and 57 (16%) reported a lifetime history of non-alcohol substance misuse. Age and gender were the main variables relevant to the extent and pattern of misuse. Self-reported non-alcohol substance misuse showed no significant relationship with a range of outcome measures. CONCLUSIONS: The high proportion of subjects reporting non-alcohol substance misuse is comparable with figures from the USA. The reports of lifetime misuse most commonly referred to cannabis, psychostimulants, LSD, opiates and anticholinergics. Misuse was concentrated in those younger than 36 years and was reported more often by males. PMID- 11731355 TI - Spontaneous lapses in dosing during chronic treatment with selective serotonin reuptake inhibitors. AB - BACKGROUND: In previous research, lapses in dosing of paroxetine or sertraline were associated with significantly more discontinuation symptoms and deterioration of depressive symptomatology compared with fluoxetine. AIMS: To evaluate dosing lapses in patients chronically treated with selective serotonin reuptake inhibitors (SSRIs) in uncontrolled circumstances. METHOD: In a prospective observational study we evaluated compliance data in chronic users of SSRIs using electronic drug exposure monitors. RESULTS: During a 3-month follow up we found that 50/69 (72.5%) patients missed at least one dosing day and 20/69 (29.0%) missed two or more consecutive days. CONCLUSIONS: About 30% of patients treated with short-acting SSRIs had dosing lapses of 2 or more days, which, as described in prior studies, is long enough to result in clinically relevant deterioration of mental status. PMID- 11731354 TI - Efficacy of usual antidepressant dosing regimens of fluoxetine in panic disorder: randomised, placebo-controlled trial. AB - BACKGROUND: Although serotonin reuptake inhibitors are effective in panic disorder, questions concerning whether doses associated with antidepressant efficacy are also effective for panic disorder remain. AIMS: To assess the efficacy of the usual antidepressant dose of fluoxetine in treating full panic attacks. METHOD: Patients with panic disorder were randomised to placebo or to fluoxetine initiated at 10 mg daily for 1 week and then increased to 20 mg daily. The trial lasted 12 weeks, but after 6 weeks patients who had failed to achieve a satisfactory response were eligible for dose escalation to a maximum of 60 mg of fluoxetine daily. RESULTS: Fluoxetine was associated with a statistically significantly greater proportion of panic-free patients compared with placebo after 6 weeks and at end-point. CONCLUSIONS: Fluoxetine at a dose of 20 mg daily is safe and efficacious in reducing symptoms of panic disorder. Patients who fail to obtain a satisfactory response at 20 mg daily may benefit from further dose increases. PMID- 11731356 TI - Long-term affective disorder in people with mild learning disability. AB - BACKGROUND: Increased risk of affective disorder in learning disability has been reported, although the extent to which this is due to adverse social and material circumstances is uncertain and there have been potential limitations in the measurement of affective disorder. AIMS: To determine risk of affective disorder in those classified with mild learning disability in the British 1946 birth cohort and to investigate whether this risk was accounted for by disadvantage in childhood and adulthood. METHOD: Learning disability was defined as the equivalent of an IQ < or =69 at age 15 years. The Present State Examination at age 36 years and the Psychiatric Symptom Frequency Scale at age 43 years provided psychiatric outcome measures. RESULTS: Learning disability was associated with a fourfold increase in risk of affective disorder, not accounted for by social and material disadvantage or by medical disorder. CONCLUSIONS: Learning disability is strongly associated with risk of affective disorder, persisting well into midlife. PMID- 11731357 TI - Outcome in consecutive emergency department attenders following a road traffic accident. AB - BACKGROUND: Little is known about the consequences of road traffic accidents. AIMS: To determine psychological and social outcome at 3 months and 1 year following a road traffic accident. METHOD: A cohort study of a 1-year sample of consecutive attenders (n=1148) aged 17-69 years at the accident and emergency department of a teaching district general hospital (excluding major head injury). Data were extracted from medica notes and from self-report at baseline, 3 months and 1 year. RESULTS: Most (61%) injuries were physically minor. At 1 year 45% reported major physical problems and 32% reported psychiatric consequences. Non injury variables were the principal predictors of outcome. CONCLUSIONS: Outcome across a range of variables is considerably worse than would be expected from the nature of the physical injuries. There is a need for changes in clinical care and socio-legal policy to prevent, identify and treat distressing and disabling chronic problems. PMID- 11731358 TI - Hidden psychiatric morbidity in elderly prisoners. AB - BACKGROUND: The number of elderly prisoners has increased significantly in Western countries over the past decade. Little is known about the psychiatric morbidity of this population. AIMS: To determine the prevalence of psychiatric morbidity in elderly sentenced prisoners. METHOD: A stratified sample of 203 male sentenced prisoners aged over 59 years, from 15 prisons in England and Wales, representing one in five men in this age group, was interviewed using semistructured standardised instruments for psychiatric illness and personality disorder. RESULTS: More than half of the elderly prisoners had a psychiatric diagnosis. The most common diagnoses were personality disorder and depressive illness. CONCLUSIONS: The prevalence of depressive illness was five times greater than that found in other studies of younger adult prisoners and elderly people in the community. Underdetected, undertreated depressive illness in elderly prisoners is an increasing public health problem. PMID- 11731359 TI - Hospital Anxiety and Depression (HAD) scale: factor structure, item analyses and internal consistency in a large population. AB - BACKGROUND: The Hospital Anxiety and Depression (HAD) rating scale is a commonly used questionnaire. Former studies have given inconsistent results as to the psychometric properties of the HAD scale. AIMS: To examine the psychometric properties of the HAD scale in a large population. METHOD: All inhabitants aged 20-89 years (n=92 100) were invited to take part in The Nord-Trondelag Health Study, Norway. A total of 65 648 subjects participated, and only completed HAD scale forms (n=51 930) formed the basis for the psychometric examinations. RESULTS: Principal component analysis extracted two factors in the HAD scale that accounted for 57% of the variance. The anxiety and depression sub-scales shared 30% of the variance. Both subscales were found to be internally consistent, with values of Cronbach's coefficient (alpha) being 0.80 and 0.76, respectively. CONCLUSIONS: Based on data from a large population, the basic psychometric properties of the HAD scale as a self-rating instrument should be considered as quite good in terms of factor structure, intercorrelation, homogeneity and internal consistency. PMID- 11731360 TI - Straight talking: an investigation of the attitudes and practice of psychoanalysts and psychotherapists in relation to gays and lesbians. AB - BACKGROUND: Early psychodynamic writing on same-gender sexual preference contributed to its pathologisation and an interest in treatment directed at changing sexual orientation. AIMS: To establish the therapeutic approaches taken by contemporary psychotherapists and psychoanalysts to gay and lesbian clients/patients. METHOD: A random sample of individuals listed as working with adults in the British Confederation of Psychotherapists' register were sent postal questionnaires. RESULTS: Data are available from 274 (69%) of 395 questionnaires. Only one of 218 respondents said that he/she was homosexual. One third said that gay and lesbian patients did have a right to a gay or lesbian therapist. A total of 179 (82% of 218) respondents described work with gay and lesbian clients/patients, and in the majority of cases sexual orientation was an important aspect of the work. CONCLUSIONS: Gays and lesbians seeking psychoanalysis or psychotherapy in the National Health Service or outside it for personal and/or training purposes will be unlikely to find a gay or lesbian therapist if they want one. The British Confederation of Psychotherapists' practitioners take on gay and lesbian clients/patients, although many do not see these social identities as relevant to the therapeutic process. Evidence from this study indicates that such clients/patients may encounter overt or covert bias, including the pathologisation of homosexuality per se. PMID- 11731361 TI - Death of neurasthenia and its psychological reincarnation: a study of neurasthenia at the National Hospital for the Relief and Cure of the Paralysed and Epileptic, Queen Square, London, 1870-1932. AB - BACKGROUND: The diagnosis of neurasthenia appeared in 1869 and rapidly became fashionable and highly prevalent. It disappeared almost completely, producing ongoing debates about what happened to the disease, which have not so far been informed by empirical data. AIMS: To use empirical historical hospital data from one specific hospital to explore several controversies about neurasthenia, including what happened to the disorder. METHOD: The annual reports of Queen Square Hospital were examined from 1870 to 1947. The prevalence of neurasthenia diagnoses as a proportion of total discharges was recorded. The possible diagnostic categories into which neurasthenia could have been reclassified were identified. Textbooks and writing by neurologists working at the hospital during this period were examined. RESULTS: Neurasthenia accounted for 6-11% of total discharges from the late 1890s to 1930, when it virtually disappeared. Men accounted for 33-50% of cases. CONCLUSIONS: Neurasthenia affected both the upper and working classes and both men and women. Neurologists, not psychiatrists, continued to see the disorder well into the 20th century. Neurasthenia did not disappear, but was reclassified into psychological diagnoses. PMID- 11731362 TI - Revisiting evolutionary psychology and psychiatry. PMID- 11731363 TI - Revisiting evolutionary psychology and psychiatry. PMID- 11731364 TI - Personality disorder: agency and responsibility. PMID- 11731365 TI - Long-term potentiation and changes seen in depression. PMID- 11731366 TI - Chromosome 22qII deletion and brain tissue composition. PMID- 11731367 TI - Leptin and antipsychotic drugs. PMID- 11731368 TI - Gender differences in treatment response to antidepressants. PMID- 11731369 TI - World psychiatric literature. PMID- 11731370 TI - Error in Andrews et al (2001). PMID- 11731373 TI - Aetiology of coronary heart disease. PMID- 11731374 TI - The "redisorganisation" of the NHS. PMID- 11731375 TI - Redefining leadership in health care. PMID- 11731376 TI - Managing intersex. PMID- 11731377 TI - The European working time directive for doctors in training. PMID- 11731378 TI - US scientists clone first human embryo. PMID- 11731379 TI - NHS to receive higher share of public spending, Brown says. PMID- 11731382 TI - AMA's new chief executive draws criticism. PMID- 11731383 TI - Mothers who kill their children can show intense grief. PMID- 11731385 TI - AIDS now fourth biggest killer worldwide, report says. PMID- 11731387 TI - Mortality in North Korea rises by 40% in seven years. PMID- 11731388 TI - Size at birth and resilience to effects of poor living conditions in adult life: longitudinal study. AB - OBJECTIVE: To determine whether men who grew slowly in utero or during infancy are more vulnerable to the later effects of poor living conditions on coronary heart disease. DESIGN: Follow up study of men for whom there were data on body size at birth and growth and social class during childhood, educational level, and social class and income in adult life. SETTING: Helsinki, Finland. PARTICIPANTS: 3676 men who were born during 1934-44, attended child welfare clinics in Helsinki, were still resident in Finland in 1971, and for whom data from the 1980 census were available. MAIN OUTCOME MEASURES: Hospital admission for or death from coronary heart disease. RESULTS: Men who had low social class or low household income in adult life had increased rates of coronary heart disease. The hazard ratio among men with the lowest annual income (<8400 pound sterling) was 1.71 (95% confidence interval 1.18 to 2.48) compared with 1.00 in men with incomes above 15, 700 pound sterling. These effects were stronger in men who were thin at birth (ponderal index <26 kg/m(3)): hazard ratio 2.58 (1.45 to 4.60) for men with lowest annual income. Among the men who were thin at birth the effects of low social class were greater in those who had accelerated weight gain between ages 1 and 12 years. Low social class in childhood further increased risk of disease, partly because it was associated with poor growth during infancy. Low educational attainment was associated with increased risk, and low income had no effect once this was taken into account. CONCLUSION: Men who grow slowly in utero remain biologically different to other men. They are more vulnerable to the effects of low socioeconomic status and low income on coronary heart disease. PMID- 11731389 TI - Measuring later health status of high risk infants: randomised comparison of two simple methods of data collection. AB - OBJECTIVE: To test two methods of providing low cost information on the later health status of survivors of neonatal intensive care. DESIGN: Cluster randomised comparison. SETTING: Nine hospitals distributed across two UK health regions. Each hospital was randomised to use one of two methods of follow up. PARTICIPANTS: All infants born 5 years) compared with controls without recurrence after long-term follow-up. Using real-time reverse transcription-PCR, we found that the normal mammary gland expressed both 17beta-HSD types 1 and 2, whereas the tumors frequently lacked detectable levels of type 2. Only 10% of the estrogen receptor positive tumors expressed type 2, whereas 31% of the ER-negative tumors did so (P = 0.031). In a case-control series of 84 patients, a high level of 17beta-HSD type 1 indicated increased risk to develop late relapse of breast cancer (odds ratio, 3.0; 95% confidence interval, 1.0-12.6; P = 0.041), whereas retained expression of type 2 indicated decreased risk (odds ratio, 0.25; 95% confidence interval, 0.05-1.2; P = 0.050). In multivariate analysis of the estrogen receptor positive patients, the absence of 17beta-HSD type 2 combined with a high expression of type 1 showed prognostic significance (P = 0.016) in addition to DNA aneuploidy (P = 0.0058), whereas progesterone receptor status did not (P = 0.71). These findings suggest that abnormal expression of 17beta-HSD isoforms has prognostic significance in breast cancer and that altered expression of these enzymes may have importance in breast cancer progression. PMID- 11731427 TI - HER-2 amplification impedes the antiproliferative effects of hormone therapy in estrogen receptor-positive primary breast cancer. AB - In experimental models, human epidermal growth factor receptor-2 (HER-2) amplification leads to estrogen independence and tamoxifen resistance in estrogen receptor (ER)-positive human breast cancer cells. Some but not all reports suggest an association between HER-2 positivity and hormone independence in breast cancer patients. This study aimed to evaluate the antiproliferative effects of endocrine therapy in HER-2-positive/ER-positive primary human breast cancer. The effect on proliferation (Ki67) of hormone therapy was assessed at 2 weeks and/or 12 weeks in biopsies from 115 primary breast cancers with ER positive tumors. The patients took part in one of 3 neoadjuvant trials of hormonal therapy with a SERM (tamoxifen or idoxifene) or an aromatase inhibitor (anastrozole or vorozole). HER-2 status was assessed by immunocytochemistry and fluorescence in situ hybridization (FISH). Fifteen patients were defined as HER-2 positive by both immunohistochemistry and FISH, with the remaining 100 patients HER-2 negative. Geometric mean Ki67 levels were substantially higher in HER-2 positive than HER-2-negative tumors (27.7% versus 11.5%, respectively; P = 0.003). In HER-2-negative patients, Ki67 was reduced by 62 and 71% at 2 and 12 weeks, respectively (P < 0.0001 for both), but HER-2-positive patients showed no significant fall. The proportional change in Ki67 was significantly different between HER-2-positive and -negative patients (P = 0.014 at 2 weeks; P = 0.047 at 12 weeks). Mean ER levels were lower in the HER-2-positive patients (P = 0.06) but the change in Ki67 was impeded even in those with high ER. Apoptotic index was reduced by 30% at 2 weeks in the HER-2-negative group. However, there were no statistically significant differences in apoptotic index between the groups. It is concluded that ER-positive/HER-2-positive primary breast carcinomas show an impeded antiproliferative response to endocrine therapy that nonetheless may vary between individual treatments. This together with high baseline proliferation is likely to translate to poor clinical response. PMID- 11731428 TI - Serologic evidence of herpes simplex virus 1 infection and oropharyngeal cancer risk. AB - In vitro and animal models suggest that the herpes simplex virus 1 (HSV1) may contribute to the development of oropharyngeal squamous cell carcinoma (OSCC). To determine whether the risk of OSCC is related to infection with HSV1 in humans, we recruited 260 patients from 18 to 65 years old who were newly diagnosed with OSCC between 1990-1995 while residing in three western Washington State counties. For comparison, we recruited at random 445 controls frequency matched to cases on age and sex. Participants completed in-person interviews and provided serum samples that were tested for antibody response to HSV1. After adjusting for sex, cigarette smoking, alcohol consumption, age, and income, HSV1 antibody positivity was associated with a slightly increased risk of OSCC [adjusted odds ratio (OR), 1.3; 95% confidence interval (CI), 0.9-2.0]. The adjusted association between HSV1 antibody positivity and OSCC risk among those who were current cigarette smokers (OR, 4.2; CI, 2.4-7.1) was stronger than would be predicted based on the additive combination of smoking alone (OR, 2.3; CI, 1.2-4.2) and HSV1 seropositivity alone (OR, 1.0; CI, 0.6-1.7). There was suggestive evidence that the association between HSV1 infection and OSCC was similarly modified by evidence of HPV infection but no evidence of effect modification with alcohol consumption. This population-based study suggests that HSV1 may enhance the development of OSCC in individuals who are already at increased risk of the disease because of cigarette smoking or HPV infection. PMID- 11731429 TI - Sensitivity to benzo(a)pyrene diol-epoxide associated with risk of breast cancer in young women and modulation by glutathione S-transferase polymorphisms: a case control study. AB - Mounting epidemiological evidence suggests that smoking may play a role in the etiology of breast cancer. Because smoking-related DNA adducts are detectable in both normal and malignant breast tissues, we hypothesized that breast cancer patients may be sensitive to tobacco-induced carcinogenesis, and this sensitivity could be modulated by variants of metabolic genes. To test this hypothesis, we evaluated benzo(a)pyrene diol-epoxide (BPDE)-induced mutagen sensitivity and polymorphisms of GSTM1 and GSTT1 in a pilot case-control study of breast cancer. Short-term cell cultures were established from blood samples of 100 female breast cancer patients and 105 healthy controls. After 5 h of in vitro exposure to 4 microM of BPDE, we harvested the lymphocytes for cytogenetic evaluation and recorded and compared the frequency of BPDE-induced chromatid breaks between cases and controls. We used a multiplex PCR-based assay to simultaneously detect polymorphisms of GSTM1 and GSTT1 from genomic DNA. We performed univariate and multivariate logistic regression analyses and calculated odds ratios (OR) and 95% confidence intervals (CIs). Cases had a significantly higher frequency of chromatid breaks than did controls (P < 0.0001). The level of chromatid breaks greater than the median value of controls was associated with a >3-fold increased risk of breast cancer [adjusted odds ratio (ORadj) = 3.11; 95% CI = 1.72-5.64]. The risk was more pronounced in those who were < 45 years (ORadj = 4.79; 95% CI = 1.87-12.3), ever-smokers (ORadj = 5.55; 95% CI = 1.85-16.6), alcohol drinkers (ORadj = 4.64; 95% CI = 1.70-12.7), and those who had the GSTT1 null variant (ORadj = 8.01; 95% CI = 1.16-55.3). These data suggest that sensitivity to BPDE induced chromosomal aberrations may contribute to the risk of developing breast cancer, and such sensitivity may be modulated by both genetic and environmental factors. Larger studies are needed to confirm our findings. PMID- 11731430 TI - RNA replicons derived from poliovirus are directly oncolytic for human tumor cells of diverse origins. AB - The failure and/or toxicity of conventional therapies for many types of human cancers underscore the need for development of safe and effective alternative treatments. Toward this goal, we describe the direct oncolytic activity of RNA based vectors derived from poliovirus, termed replicons, which are genetically incapable of producing infectious virus. These replicons are cytopathic in vitro for human tumor cells originating from brain, breast, lung, ovary, and skin (melanoma). The cytopathic effects in a malignant glioma cell line were associated with nuclear DNA condensation, indicative of cells undergoing apoptosis. Injection of replicons into established xenograft flank tumors in scid mice resulted in oncolytic activity and extended survival. Inoculation of replicons into established intracranial xenograft tumors in scid mice resulted in tumor infection within 8 h and extended survival. Histological analysis revealed that replicons had infected tumor cells at the site of inoculation and, most importantly, diffused to infect tumor cells that had metastasized from the initial site of implantation. The wide spectrum of cytopathic activity for human tumors combined with effective distribution after in vivo inoculation establishes the therapeutic potential of poliovirus replicons for a variety of cancers. PMID- 11731431 TI - Inhibition of angiogenesis and metastasis in two murine models by the matrix metalloproteinase inhibitor, BMS-275291. AB - BMS-275291 is an p.o. bioavailable, sulfhydryl-based matrix metalloproteinase (MMP) inhibitor currently in clinical development for the treatment of cancer. This inhibitor was designed to potently inhibit MMP activities while minimally affecting those of other metalloproteases (e.g., sheddases) involved in the release of cell-associated molecules such as tumor necrosis factor-alpha, tumor necrosis factor-alpha receptor, interleukin-6 receptor, or L-selectin. In vitro, BMS-275291 is a potent inhibitor (nM) of the activities of MMP-1, MMP-2, MMP-7, MMP-9, and MMP-14. BMS-275291 inhibits tumor growth in a B16BL6 model of experimental metastasis, and in this model, BMS-275291 treatment results in a dose-dependent reduction in the number of lung metastases compared with vehicle controls. BMS-275291 also inhibits angiogenesis in a murine angiogenesis model, where once daily treatment with BMS-275291 results in a dose-dependent inhibition of endothelial cell migration into s.c. implanted Matrigel plugs. Pharmacokinetic studies demonstrated that the plasma concentrations of parent BMS-275291 in mice exceeds the in vitro IC(50) values for MMP-1, MMP-2, MMP-7, MMP-9, and MMP-14 for at least 4 h after the administration of a therapeutic dose of BMS-275291. Taken together, these data demonstrate that BMS-275291 inhibits MMP activities that contribute to tumor metastasis and angiogenesis. PMID- 11731432 TI - Antisense oligodeoxynucleotide targeted to Midkine, a heparin-binding growth factor, suppresses tumorigenicity of mouse rectal carcinoma cells. AB - Midkine (MK), a heparin-binding growth factor, is overexpressed in a wide range of human carcinomas and is believed to contribute to tumorigenesis and tumor progression. To develop an antitumor reagent, we designed a phosphorothioate antisense oligodeoxynucleotide molecule based on the secondary structure of MK mRNA. The antisense MK at the dosage of 5 microM suppressed MK production by CMT 93 mouse rectal carcinoma cells after cationic liposome-mediated transfection, to 13% of that in control cultures. The growth of CMT-93 cells and their colony formation in soft agar were inhibited by the addition of the antisense MK, whereas the control reagent, the sense MK, showed no effects. On s.c. injection into nude mice, CMT-93 cells transfected with the antisense MK formed tumors much smaller than those by control cells. Finally, untreated CMT-93 cells were inoculated to nude mice, and 7 days later the antisense MK (50 microM) with atelocollagen was directly injected into the preformed tumor region to evaluate the curative effect; the injection was repeated at the interval of 2 weeks. During the period of 10-41 days after initiation of therapy, the rate of increase of tumor volume treated with the antisense MK was found to be about 4.2-fold lower than that seen after treatment with the sense MK. On this occasion, proliferation of tumor cells as estimated by 5-bromodeoxyuridine incorporation was strongly inhibited, whereas angiogenesis was less affected. These findings strongly suggested the usefulness of MK antisense oligodeoxynucleotide as a new reagent for cancer therapy. PMID- 11731433 TI - The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces differentiation of human breast cancer cells. AB - Histone deacetylase (HDACs) regulate histone acetylation by catalyzing the removal of acetyl groups on the NH(2)-terminal lysine residues of the core nucleosomal histones. Modulation of the acetylation status of core histones is involved in the regulation of the transcriptional activity of certain genes. HDAC activity is generally associated with transcriptional repression. Aberrant recruitment of HDAC activity has been associated with the development of certain human cancers. We have developed a class of HDAC inhibitors, such as suberoylanilide hydroxamic acid (SAHA), that were initially identified based on their ability to induce differentiation of cultured murine erythroleukemia cells. Additional studies have demonstrated that SAHA inhibits the growth of tumors in rodents. In this study we have examined the effects of SAHA on MCF-7 human breast cancer cells. We found that SAHA causes the inhibition of proliferation, accumulation of cells in a dose-dependent manner in G(1) then G(2)-M phase of the cell cycle, and induction of milk fat globule protein, milk fat membrane globule protein, and lipid droplets. Growth inhibition was associated with morphological changes including the flattening and enlargement of the cytoplasm, and a decrease in the nuclear:cytoplasmic ratio. Withdrawal of SAHA led to reentry of cells into the cell cycle and reversal to a less differentiated phenotype. SAHA induced differentiation in the estrogen receptor-negative cell line SKBr-3 and the retinoblastoma-negative cell line MDA-468. We propose that SAHA has profound antiproliferative activity by causing these cells to undergo cell cycle arrest and differentiation that is dependent on the presence of SAHA. SAHA and other HDAC inhibitors are currently in Phase I clinical trials. These findings may impact the clinical use of these drugs. PMID- 11731434 TI - MIG (CXCL9) chemokine gene therapy combines with antibody-cytokine fusion protein to suppress growth and dissemination of murine colon carcinoma. AB - The induction of a CTL response capable of eradicating disseminated tumor metastases and the establishment of a persistent tumor-protective immunity remain major goals of cancer immunotherapy. Here, we demonstrate for the first time that the combination of interleukin 2 (IL-2) targeted to the tumor microenvironment by a recombinant antibody-IL-2 fusion protein (huKS1/4-IL-2) with gene therapy by the murine chemokine MIG (CXCL9) markedly reduced s.c. tumor burden and decisively suppressed dissemination of experimental lung metastases of CT26-KSA colon carcinoma in syngeneic BALB/c mice. This combined therapy significantly prolonged the life span of these mice 3-4-fold by concurrently delivering MIG and IL-2 to the tumor site and thereby achieving chemoattraction of T cells together with their activation. The antitumor effect obtained was mediated predominantly by MHC class I antigen-restricted CD8(+) T cells with help from MHC class II antigen-restricted CD4(+) T lymphocytes. In addition, the MIG chemokine also induced angiostatic effects in the tumor vasculature. Taken together, this combination of MIG chemokine gene therapy with tumor-targeted cytokine IL-2 provides an approach for the rational design of novel cancer immunotherapy modalities. PMID- 11731435 TI - Retinoid-related molecules induce cytochrome c release and apoptosis through activation of c-Jun NH(2)-terminal kinase/p38 mitogen-activated protein kinases. AB - Retinoid-related molecules have been described that induce apoptosis in a variety of cancer cell lines. Of particular interest is the apoptotic activity of the all trans-retinoic acid receptor gamma-selective molecules MX2870-1 and MX3350-1. These compounds have been shown to be effective in vivo against lung cancer and could therefore serve as important leads for novel anticancer drugs. We analyzed the death signaling pathways activated by these molecules. We observed that apoptotic retinoid-related molecules (RRMs) cause the release of cytochrome c from the mitochondria and subsequent activation of caspases 9 and 3. This was preceded by a strong and sustained activation of c-Jun NH(2)-terminal kinase as well as p38 kinase, which was independent of caspase activity. Inhibition of p38 kinase activity by the specific inhibitor SB203580 did not affect the induction of apoptosis by MX2870-1. However, interference with the activation of c-Jun NH(2)-terminal kinase and p38 stress kinases by PD169316 completely blocked all signs of apoptosis, including caspase activity, DNA fragmentation, and phosphatidylserine externalization. PD169316 also prevented the cleavage of Bid and the release of cytochrome c induced by this class of RRMs. Furthermore, processing and activation of different caspases by MX2870-1 was completely inhibited by increasing concentrations of PD169316. Thus, the investigated RRMs induce a death pathway, which is independent of Fas ligand, that is also activated by UV radiation and other agents. Our findings open the possibility for the future use of this class of RRMs in combination therapies with other anticancer drugs. PMID- 11731436 TI - Vaccination of pediatric solid tumor patients with tumor lysate-pulsed dendritic cells can expand specific T cells and mediate tumor regression. AB - Dendritic cells (DCs) have been shown to be a promising adjuvant for inducing immunity to cancer. We evaluated tumor lysate-pulsed DC in a Phase I trial of pediatric patients with solid tumors. Children with relapsed solid malignancies who had failed standard therapies were eligible. The vaccine used immature DC (CD14-, CD80+, CD86+, CD83-, and HLA-DR+) generated from peripheral blood monocytes in the presence of granulocyte/monocyte colony-stimulating factor and interleukin-4. These DC were then pulsed separately with tumor cell lysates and the immunogenic protein keyhole limpet hemocyanin (KLH) for 24 h and then combined. A total of 1 x 10(6) to 1 x 10(7) DC are administered intradermally every 2 weeks for a total of three vaccinations. Fifteen patients (ages 3-17 years) were enrolled with 10 patients completing all vaccinations. Leukapheresis yields averaged 2.8 x 10(8) peripheral blood mononuclear cells (PBMC)/kg, and DC yields averaged 10.9% of starting PBMC. Patients with neuroblastoma, sarcoma, and renal malignancies were treated without obvious toxicity. Delayed-type hypersensitivity (DTH) response was detected in 7 of 10 patients for KLH and 3 of 6 patients for tumor lysates. Priming of T cells to KLH was seen in 6 of 10 patients and to tumor in 3 of 7 patients as demonstrated by specific IFN-gamma secreting T cells in unstimulated PBMCs. Significant regression of multiple metastatic sites was seen in 1 patient. Five patients showed stable disease, including 3 who had minimal disease at time of vaccine therapy and remain free of tumor with 16-30 months follow-up. Our results demonstrate that it is feasible to generate large numbers of functional DC from pediatric patients even in those highly pretreated and with a large tumor burden. The DC can be administered in an outpatient setting without any observable toxicity. Most importantly, we have demonstrated the ability of the tumor lysate/KLH-pulsed DC to generate specific T cell responses and to elicit regression of metastatic disease. PMID- 11731437 TI - A new mouse model for evaluating the immunotherapy of human colorectal cancer. AB - A new murine model of human colorectal cancer was generated by crossing human carcinoembryonic antigen (CEA) transgenic mice (H-2K(b)) with adenomatous polyposis coli (Apc1638N) knockout mice (H-2K(b)). The resulting hybrid mice developed gastrointestinal polyps in 6-8 months that progressed to invasive carcinomas with a similar pattern of dysplasia and CEA expression as observed in human colorectal cancer. These animals exhibited incomplete or partial tolerance to CEA as evidenced by delayed growth of CEA-expressing tumors and the inability to inhibit CEA-specific CTL responses. These results have important implications for understanding the role of CEA-specific immunity in human colon cancer patients and suggest that vaccine strategies targeting CEA may be feasible. This model provides a powerful system for evaluating antigen-specific tumor immunity against spontaneous tumors arising in an orthotopic location and permits evaluation of therapeutic vaccine strategies for human colorectal cancer. PMID- 11731438 TI - Aberrations in the fragile histidine triad (FHIT) gene in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) seems to be closely associated with lung carcinogenesis. To identify the genetic characteristics of precancerous IPF lesions in the peripheral lung, we performed PCR-based microsatellite analysis with DNA extracted from microdissected tissues; fluorescent in situ hybridization (FISH) analysis of the fragile histidine triad (FHIT) gene and immunohistochemical analysis of Fhit protein expression in samples of metaplasias and bronchiolar epithelia obtained from patients with IPF. We used four microsatellite markers of the FHIT gene within or flanking the FHIT gene on chromosome 3p for loss of heterozygosity (LOH) analysis. LOH of the FHIT locus was frequently found among the lesions of metaplasias and bronchiolar epithelia in the patients with IPF [62 (52%) of 119 informative lesions]. Fifty-four (73%) of the 74 lesions of metaplasias and bronchiolar epithelia obtained from the IPF patients with lung carcinoma and 8 (17%) of the 46 samples obtained from the IPF patients without lung carcinoma showed LOH at the FHIT gene (P < 0.0001). We confirmed allelic loss in the metaplasias and bronchiolar epithelia of IPF by FISH analysis of the FHIT gene. Additionally, the level of Fhit protein expression in the metaplastic cells of IPF was frequently reduced. Our findings suggest that allelic loss of the FHIT gene may be involved in carcinogenesis in the peripheral lung of patients with IPF. PMID- 11731439 TI - Novel target genes of the Ah (dioxin) receptor: transcriptional induction of N myristoyltransferase 2. AB - Dioxins are potent mammalian carcinogens and toxins affecting liver, skin, and immune and reproductive systems. The intracellular Ah receptor, a ligand dependent transcription factor of the basic region/helix-loop-helix/Per-Ahr/Arnt Sim homology domain (bHLH-PAS) protein family, mediates responses to dioxins. Target genes of the Ah receptor that mediate dioxin toxicity and carcinogenicity are, however, mostly unknown. We used 5L rat hepatoma cells to identify dioxin inducible genes by suppression subtractive hybridization. Eleven cDNA fragments were identified that represented novel sequences or genes for which induction by dioxins had not been known. N-myristoyltransferase 2 (NMT2) is one of the later dioxin-inducible genes. Induction of NMT2 was confirmed in livers of mice in vivo. NMT2 induction was a direct consequence of Ah receptor activation in 5L cells. [(3)H]myristic acid incorporation into 5L cell proteins was inducible by dioxins, indicating that protein myristoylation is a regulated rather than a housekeeping function and that NMT activity is limiting in noninduced 5L cells. Here we show for the first time that expression of NMT2 and induced protein myristoyltransferase activity are direct responses to carcinogen exposure. Because inappropriate protein NH(2)-terminal myristoylation appears to play a role in carcinogenesis, induction of NMT2 may play a central role in dioxin carcinogenicity. PMID- 11731440 TI - Identification and characterization of differentially methylated CpG islands in pancreatic carcinoma. AB - To identify CpG islands differentially methylated in pancreatic adenocarcinoma, we used methylated CpG island amplification (MCA) coupled with representational difference analysis. Of 42 CpG islands identified by MCA/representational difference analysis, 7 CpG islands [methylated in carcinoma of the pancreas (MICP)] were differentially methylated in a panel of eight pancreatic cancer cell lines compared with normal pancreas. In a larger panel of 75 pancreatic adenocarcinomas, these 7 MICPs (ppENK, Cyclin G, ZBP, MICP25, 27, 36, and 38) were methylated in 93, 3, 9, 15, 48, 19, and 41% of cancers, respectively, by methylation-specific PCR but not in any of 15 normal pancreata. In pancreatic cancer cell lines, methylation of ppENK, a gene with known growth suppressive properties, was associated with transcriptional silencing that was reversible with 5-aza-2'-deoxycytidine treatment. Relationships between the methylation patterns of pancreatic adenocarcinomas and their clinicopathological features were also determined. Larger pancreatic cancers and those from older patients (P = 0.017) harbored more methylated loci than smaller tumors and those from younger patients (P = 0.017). ppENK, MICP25, and 27 were variably methylated in normal gastric, duodenal, and colonic mucosae. These data indicate that aberrant methylation of ppENK and its transcriptional repression is a common event in pancreatic carcinogenesis. PMID- 11731441 TI - Molecular analysis of single colonies reveals a diverse origin of initial clonal proliferation in B-precursor acute lymphoblastic leukemia that can precede the t(12;21) translocation. AB - The pathogenesis of pediatric B-precursor acute lymphoblastic leukemia is largely unknown, and even with nonrandom chromosomal translocations present, the precise order of clonal molecular events is undefined. We developed an in vitro system using cytokines interleukin (IL)-3, IL-7, IL-10, and FMS-like tyrosine kinase 3 ligand with CD40 ligand-expressing fibroblasts to obtain single blast colonies from which clonal immunoglobulin heavy chain (IgH), T-cell receptor delta gene rearrangements, and, in t(12;21)-positive cases, TEL-AML1 fusion transcripts could be simultaneously PCR amplified. The proliferation of early tumor progenitors increased subclone detection enabling us, in seven diagnostic samples, to determine the stage of differentiation at which each leukemia occurred. Four were derived from the stage before initiation of IgH rearrangement, one during recombination of variable, joining, and diversity segments of the heavy chain gene VDJ(H), and two after completion of IgH rearrangement. Furthermore, analysis of a t(12;21)-positive leukemia with unusually late onset, identified both TEL-AML1-positive and -negative colonies carrying a clonal T-cell receptor delta rearrangement, inferring the presence of clonal expansion before the occurrence of the t(12;21). In contrast, in a typical, early onset t(12;21)-positive leukemia, the t(12;21) appeared to be the first clonal event. In both leukemias, the t(12;21) occurred before recombination of variable, joining and diversity segments of the heavy chain gene VDJ. PMID- 11731442 TI - Replication protein A2 phosphorylation after DNA damage by the coordinated action of ataxia telangiectasia-mutated and DNA-dependent protein kinase. AB - Replication protein A (RPA, also known as human single-stranded DNA-binding protein) is a trimeric, multifunctional protein complex involved in DNA replication, DNA repair, and recombination. Phosphorylation of the RPA2 subunit is observed after exposure of cells to ionizing radiation (IR) and other DNA damaging agents, which implicates the modified protein in the regulation of DNA replication after DNA damage or in DNA repair. Although ataxia telangiectasia mutated (ATM) and DNA-dependent protein kinase (DNA-PK) phosphorylate RPA2 in vitro, their role in vivo remains uncertain, and contradictory results have been reported. Here we show that RPA2 phosphorylation is delayed in cells deficient in one of these kinases and completely abolished in wild-type, ATM, or DNA-PK deficient cells after treatment with wortmannin at a concentration-inhibiting ATM and DNA-PK. Caffeine, an inhibitor of ATM and ATM-Rad3 related (ATR) but not DNA PK, generates an ataxia-telangiectasia-like response in wild-type cells, prevents completely RPA2 phosphorylation in DNA-PKcs deficient cells, but has no effect on ataxia-telangiectasia cells. These observations rule out ATR and implicate both ATM and DNA-PK in RPA2 phosphorylation after exposure to IR. UCN-01, an inhibitor of protein kinase C, Chk1, and cyclin-dependent kinases, has no effect on IR induced RPA2 phosphorylation. Because UCN-01 abrogates checkpoint responses, this observation dissociates RPA2 phosphorylation from checkpoint activation. Phosphorylated RPA has a higher affinity for nuclear structures than unphosphorylated RPA suggesting functional alterations in the protein. In an in vitro assay for DNA replication, DNA-PK is the sole kinase phosphorylating RPA2, indicating that processes not reproduced in the in vitro assay are required for RPA2 phosphorylation by ATM. Because RPA2 phosphorylation kinetics are distinct from those of the S phase checkpoint, we propose that DNA-PK and ATM cooperate to phosphorylate RPA after DNA damage to redirect the functions of the protein from DNA replication to DNA repair. PMID- 11731443 TI - Transient expression of cyclin D1 is sufficient to promote hepatocyte replication and liver growth in vivo. AB - Cyclin D1 regulates mitogen-dependent progression through G(1) phase in cultured cells, and its overexpression in malignant cells is thought to contribute to autonomous proliferation in vivo. However, previous studies in cell lines have not demonstrated that cyclin D1 is sufficient to trigger cell replication. In this study, we found that transient transfection of adult hepatocytes with cyclin D1 stimulated assembly of active cyclin D1/cdk4 complexes, robust hepatocyte proliferation, and liver growth in the intact animal. After several days, hepatocyte proliferation was inhibited despite the persistence of high levels of cyclin D1 and cyclin E, suggesting that endogenous antiproliferative mechanisms were induced. Our data suggest that this antiproliferative response includes the marked up-regulation of p21, which in turn inhibits cyclin D1/cdk4 and cyclin E/cdk2 complexes. This study offers further evidence that cyclin D1 plays a pivotal role in the regulation of hepatocyte proliferation in the liver. Furthermore, this model may offer a unique system to study the normal cellular response to cyclin D1 expression in vivo. PMID- 11731444 TI - Vascular endothelial growth factor isoforms display distinct activities in promoting tumor angiogenesis at different anatomic sites. AB - The gene for the major angiogenic factor, vascular endothelial growth factor (VEGF), encodes several spliced isoforms. We reported previously that overexpression of two VEGF isoforms, VEGF(121) and VEGF(165), by human glioma U87 MG cells induced tumor-associated intracerebral hemorrhage, whereas expression of a third form, VEGF(189), did not cause vessel rupture. Here, we test whether these VEGF isoforms have distinct activities for enhancing vascularization and growth of gliomas in mice. U87 MG cells that overexpressed VEGF(165) or VEGF(189) grew more rapidly than the parental cells in both s.c. and intracranial (i.c.) locations. However, cells that overexpressed VEGF(121) only showed enhancement of i.c. tumor growth but had a minimal effect on s.c. glioma progression. At both anatomical sties, VEGF(165) and VEGF(189) strongly augmented neovascularization, whereas VEGF(121) only increased vessel density in brain tumors. In each type of glioma, expression of VEGF receptors -1 and -2 largely phenocopied the tumor vasculature, because increased VEGF/VEGF receptor-activated microvessel densities were strongly correlated with the angiogenicity and tumorigenicity elicited by the VEGF isoforms at both anatomical sites. One notable difference between the sites was the expression of vitronectin, a prototypic ligand of alpha(v)beta(3) and alpha(v)beta(5) integrins, detected in i.c. but not in s.c., gliomas. Endothelial cell migration stimulated by VEGF(121) was potentiated by vitronectin to a greater extent than that stimulated by VEGF(165). This data demonstrates that VEGF isoforms have distinct activities at different anatomical sites and suggest that the microenvironment of different tissues affects the function of VEGF isoforms. PMID- 11731445 TI - Differential expression of manganese superoxide dismutase and catalase in lung cancer. AB - Reactive oxygen species (ROS) are important in the initiation and promotion of cells to neoplastic growth. In this context, cigarette smoke exposure, the primary risk factor in lung cancer development, leads to high levels of ROS within the human airway. Although well-equipped with an integrated antioxidant defense system consisting of low-molecular weight antioxidants such as glutathione and intracellular enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase, the lungs are vulnerable to increased endogenous and exogenous oxidative insults. Antioxidants increase in response to oxidative stress and minimize ROS-induced injury in experimental systems, indicating that antioxidant levels may determine whether ROS can initiate lung carcinogenesis. On this basis, we hypothesized that antioxidants would be decreased in lung carcinoma cells as compared with tumor-free adjacent lung tissues. Antioxidant expression was evaluated in 16 lung tumor and 21 tumor-free lung tissues collected between the years 1993 and 2001 from 24 individuals with surgically resectable non-small cell lung cancer, i.e., adenocarcinoma and squamous cell carcinoma. Total SOD activity was increased (P = 0.035), catalase activity decreased (P = 0.002), and glutathione and glutathione peroxidase were similar in tumors compared with tumor-free lung tissues. Alterations in antioxidant activities were attributable to increased manganese SOD and decreased catalase protein and mRNA expression in tumors. Immunohistochemical localization of catalase in the lung revealed decreased or no expression in the tumor cells, although healthy adjacent airway epithelial cells were strongly positive for catalase. Parallel changes in antioxidant activities, protein, and mRNA expression were noted in A549 lung carcinoma cell lines exposed to cytokines (tumor necrosis factor-alpha, interleukin 1beta, and IFN-gamma). Thus, inflammation in the lung may contribute to high levels of manganese SOD and decreased catalase, which together may lead to increased hydrogen peroxide intracellularly and create an intracellular environment favorable to DNA damage and the promotion of cancer. PMID- 11731446 TI - Interference of tenascin-C with syndecan-4 binding to fibronectin blocks cell adhesion and stimulates tumor cell proliferation. AB - Tenascin-C is an adhesion-modulatory extracellular matrix molecule that is highly expressed in tumors. To investigate the effect of tenascin-C on tumor cells, we analyzed its antiadhesive nature and effect on tumor cell proliferation in a fibronectin context. Glioblastoma and breast carcinoma cell adhesion was compromised by a mixed fibronectin/tenascin-C substratum, which concomitantly caused increased tumor-cell proliferation. We identified the antiadhesive mechanism as a specific interference of tenascin-C with cell binding to the HepII/syndecan-4 site in fibronectin through direct binding of tenascin-C to the 13th fibronectin type III repeat (FNIII13). Cell adhesion and proliferation levels were restored by the addition of FNIII13. Overexpression of syndecan-4, but not syndecan-1 or -2, reverted the cell adhesion defect of tenascin-C. We characterized FNIII13 as the binding site for syndecan-4. Thus we describe a novel mechanism by which tenascin-C impairs the adhesive function of fibronectin through binding to FNIII13, thereby inhibiting the coreceptor function of syndecan-4 in fibronectin-induced integrin signaling. PMID- 11731447 TI - Correspondence re: F. Coulet et al., Detection of plasma tumor DNA in head and neck squamous cell carcinoma by microsatellite typing and p53 mutation analysis. Cancer Res., 60: 707-709, 2000. PMID- 11731448 TI - EphA4/ephrin-A5 interactions in muscle precursor cell migration in the avian forelimb. AB - Limb muscles derive from muscle precursor cells that lie initially in the lateral portion of the somitic dermomyotome and subsequently migrate to their target limb regions, where muscle-specific gene transcription is initiated. Although several molecules that control the generation and delamination of muscle precursor cells have been identified, little is known about the mechanisms that guide muscle precursor cell migration in the limb. We have examined the distribution of members of the Eph family during muscle precursor cell development. The EphA4 receptor tyrosine kinase and its ligand, ephrin-A5, are expressed by muscle precursor cells and forelimb mesoderm in unique spatiotemporal patterns during the period when muscle precursors delaminate from the dermomyotome and migrate into the limb. To test the function of EphA4/ephrin-A5 interactions in muscle precursor migration, we used targeted in ovo electroporation to express ephrin-A5 ectopically specifically in the presumptive limb mesoderm. In the presence of ectopic ephrin-A5, Pax7-positive muscle precursor cells are significantly reduced in number in the proximal limb, compared with controls, and congregate abnormally near the lateral dermomyotome. In stripe assays, isolated muscle precursor cells avoid substrate-bound ephrin-A5 and this avoidance is abolished by addition of soluble ephrin-A5. These data suggest that ephrin-A5 normally restricts migrating, EphA4-positive muscle precursor cells to their appropriate territories in the forelimb, disallowing entry into abnormal embryonic regions. PMID- 11731449 TI - A subtilisin-like serine protease is required for epidermal surface formation in Arabidopsis embryos and juvenile plants. AB - The surfaces of land plants are covered with a cuticle that is essential for retention of water. Epidermal surfaces of Arabidopsis thaliana embryos and juvenile plants that were homozygous for abnormal leaf shape1 (ale1) mutations were defective, resulting in excessive water loss and organ fusion in young plants. In ale1 embryos, the cuticle was rudimentary and remnants of the endosperm remained attached to developing embryos. Juvenile plants had a similar abnormal cuticle. The ALE1 gene was isolated using a transposon-tagged allele ale1-1. The predicted ALE1 amino acid sequence was homologous to those of subtilisin-like serine proteases. The ALE1 gene was found to be expressed within certain endosperm cells adjacent to the embryo and within the young embryo. Expression was not detected after germination. Our results suggest that the putative protease ALE1 affects the formation of cuticle on embryos and juvenile plants and that an appropriate cuticle is required for separation of the endosperm from the embryo and for prevention of organ fusion. PMID- 11731450 TI - Dpp and Hh signaling in the Drosophila embryonic eye field. AB - We have analyzed the function of the Decapentaplegic (Dpp) and Hedgehog (Hh) signaling pathways in partitioning the dorsal head neurectoderm of the Drosophila embryo. This region, referred to as the anterior brain/eye anlage, gives rise to both the visual system and the protocerebrum. The anlage splits up into three main domains: the head midline ectoderm, protocerebral neurectoderm and visual primordium. Similar to their vertebrate counterparts, Hh and Dpp play an important role in the partitioning of the anterior brain/eye anlage. Dpp is secreted in the dorsal midline of the head. Lowering Dpp levels (in dpp heterozygotes or hypomorphic alleles) results in a 'cyclops' phenotype, where mid dorsal head epidermis is transformed into dorsolateral structures, i.e. eye/optic lobe tissue, which causes a continuous visual primordium across the dorsal midline. Absence of Dpp results in the transformation of both dorsomedial and dorsolateral structures into brain neuroblasts. Regulatory genes that are required for eye/optic lobe fate, including sine oculis (so) and eyes absent (eya), are turned on in their respective domains by Dpp. The gene zerknuellt (zen), which is expressed in response to peak levels of Dpp in the dorsal midline, secondarily represses so and eya in the dorsomedial domain. Hh and its receptor/inhibitor, Patched (Ptc), are expressed in a transverse stripe along the posterior boundary of the eye field. As reported previously, Hh triggers the expression of determinants for larval eye (atonal) and adult eye (eyeless) in those cells of the eye field that are close to the Hh source. Eya and So, which are induced by Dpp, are epistatic to the Hh signal. Loss of Ptc, as well as overexpression of Hh, results in the ectopic induction of larval eye tissue in the dorsal midline (cyclopia). We discuss the similarities between vertebrate systems and Drosophila with regard to the fate map of the anterior brain/eye anlage, and its partitioning by Dpp and Hh signaling. PMID- 11731452 TI - Populations of NGF-dependent neurones differ in their requirement for BAX to undergo apoptosis in the absence of NGF/TrkA signalling in vivo. AB - Reports that apoptosis within populations of neurotrophin-dependent neurones is virtually eliminated in BAX-deficient mice and that BAX-deficient neurones survive indefinitely in culture without neurotrophins have led to the view that BAX is required for the death of neurotrophin-deprived neurones. To further examine this assertion in vivo, we have studied two populations of NGF-dependent neurones during the period of naturally occurring neuronal death in mice that lack BAX, NGF or the NGF receptor TrkA, alone and in combination. In the superior cervical ganglion (SCG), naturally occurring neuronal death and the massive loss of neurones that took place in the absence of NGF or TrkA were completely prevented by elimination of BAX. However, in the trigeminal ganglion, naturally occurring neuronal death was only partly abrogated by the elimination of BAX, and although the massive neuronal death that took place in this ganglion in the absence of NGF or TrkA was initially delayed in embryos lacking BAX, this subsequently occurred unabated. Accordingly, BAX-deficient neurones survived in defined without NGF whereas BAX-deficient trigeminal neurones died in the absence of NGF. These results indicate that whereas BAX is required for the death of SCG neurones during normal development and when these neurones are deprived of NGF/TrkA signalling in vivo, the death of trigeminal ganglion neurones occurs independently of BAX when they are deprived of NGF/TrkA signalling. We conclude that BAX is not universally required for neuronal death induced by neurotrophin deprivation, but that there are major differences for the requirement for BAX among different populations of NGF-dependent neurones. PMID- 11731451 TI - The Drosophila daughterless gene autoregulates and is controlled by both positive and negative cis regulation. AB - As the only class I helix-loop-helix transcription factor in Drosophila, Daughterless (Da) has generally been regarded as a ubiquitously expressed binding partner for other developmentally regulated bHLH transcription factors. From analysis of a novel tissue-specific allele, da(lyh), we show that da expression is not constitutive, but is dynamically regulated. This transcriptional regulation includes somatic ovary-specific activation, autoregulation and negative regulation. Unexpectedly, the diverse functions of da may require that expression levels be tightly controlled in a cell and/or tissue-specific manner. Our analysis of da(lyh) identifies it as the first springer insertion that functions as an insulating element, with its disruptive activity mediated by the product of a fourth chromosome gene, Suppressor of lyh [Su(lyh)]. PMID- 11731453 TI - The protein kinase Pelle mediates feedback regulation in the Drosophila Toll signaling pathway. AB - Dorsoventral polarity in the Drosophila embryo is established through a signal transduction cascade triggered in ventral and ventrolateral regions. Activation of a transmembrane receptor, Toll, leads to localized recruitment of the adaptor protein Tube and protein kinase Pelle. Signaling through these components directs degradation of the IkappaB-like inhibitor Cactus and nuclear translocation of the Rel protein Dorsal. Here we show through confocal immunofluorescence microscopy that Pelle functions to downregulate the signal-dependent relocalization of Tube. Inactivation of the Pelle kinase domain, or elimination of the Tube-Pelle interaction, dramatically increases Tube recruitment to the ventral plasma membrane in regions of active signaling. We also characterize a large collection of pelle alleles, identifying the molecular lesions in these alleles and their effects on Pelle autophosphorylation, Tube phosphorylation and Tube relocalization. Our results point to a mechanism operating to modulate the domain or duration of signaling downstream from Tube and Pelle. PMID- 11731454 TI - Notch-Delta signaling induces a transition from mitotic cell cycle to endocycle in Drosophila follicle cells. AB - In many developmental processes, polyploid cells are generated by a variation of the normal cell cycle called the endocycle in which cells increase their genomic content without dividing. How the transition from the normal mitotic cycle to endocycle is regulated is poorly understood. We show that the transition from mitotic cycle to endocycle in the Drosophila follicle cell epithelium is regulated by the Notch pathway. Loss of Notch function in follicle cells or its ligand Delta function in the underlying germline disrupts the normal transition of the follicle cells from mitotic cycle to endocycle, mitotic cycling continues, leading to overproliferation of these cells. The regulation is at the transcriptional level, as Su(H), a downstream transcription factor in the pathway, is also required cell autonomously in follicle cells for proper transitioning to the endocycle. One target of Notch and Su(H) is likely to be the G2/M cell cycle regulator String, a phosphatase that activates Cdc2 by dephosphorylation. String is normally repressed in the follicle cells just before the endocycle transition, but is expressed when Notch is inactivated. Analysis of the activity of String enhancer elements in follicle cells reveals the presence of an element that promotes expression of String until just before the onset of polyploidy in wild-type follicle cells but well beyond this stage in Notch mutant follicle cells. This suggests that it may be the target of the endocycle promoting activity of the Notch pathway. A second element that is insensitive to Notch regulation promotes String expression earlier in follicle cell development, which explains why Notch, while active at both stages, represses String only at the mitotic cycle-endocycle transition. PMID- 11731455 TI - Regulation of ureteric bud outgrowth by Pax2-dependent activation of the glial derived neurotrophic factor gene. AB - The outgrowth of the ureteric bud from the posterior nephric duct epithelium and the subsequent invasion of the bud into the metanephric mesenchyme initiate the process of metanephric, or adult kidney, development. The receptor tyrosine kinase RET and glial cell-derived neurotrophic factor (GDNF) form a signaling complex that is essential for ureteric bud growth and branching morphogenesis of the ureteric bud epithelium. We demonstrate that Pax2 expression in the metanephric mesenchyme is independent of induction by the ureteric bud. Pax2 mutants are deficient in ureteric bud outgrowth and do not express GDNF in the uninduced metanephric mesenchyme. Furthermore, Pax2 mutant mesenchyme is unresponsive to induction by wild-type heterologous inducers. In normal embryos, GDNF is sufficient to induce ectopic ureter buds in the posterior nephric duct, a process inhibited by bone morphogenetic protein 4. However, GDNF replacement in organ culture is not sufficient to stimulate ureteric bud outgrowth from Pax2 mutant nephric ducts, indicating additional defects in the nephric duct epithelium of Pax2 mutants. Pax2 can activate expression of GDNF in cell lines derived from embryonic metanephroi. Furthermore, Pax2 protein can bind to upstream regulatory elements within the GDNF promoter region and can transactivate expression of reporter genes. Thus, activation of GDNF by Pax2 coordinates the position and outgrowth of the ureteric bud such that kidney development can begin. PMID- 11731456 TI - The Snail protein family regulates neuroblast expression of inscuteable and string, genes involved in asymmetry and cell division in Drosophila. AB - Delaminated neuroblasts in Drosophila function as stem cells during embryonic central nervous system development. They go through repeated asymmetric divisions to generate multiple ganglion mother cells, which divide only once more to produce postmitotic neurons. Snail, a zinc-finger transcriptional repressor, is a pan-neural protein, based on its extensive expression in neuroblasts. Previous results have demonstrated that Snail and related proteins, Worniu and Escargot, have redundant and essential functions in the nervous system. We show that the Snail family of proteins control central nervous system development by regulating genes involved in asymmetry and cell division of neuroblasts. In mutant embryos that have the three genes deleted, the expression of inscuteable is significantly lowered, while the expression of other genes that participate in asymmetric division, including miranda, staufen and prospero, appears normal. The deletion mutants also have much reduced expression of string, suggesting that a key component that drives neuroblast cell division is abnormal. Consistent with the gene expression defects, the mutant embryos lose the asymmetric localization of prospero RNA in neuroblasts and lose the staining of Prospero protein that is normally present in ganglion mother cells. Simultaneous expression of inscuteable and string in the snail family deletion mutant efficiently restores Prospero expression in ganglion mother cells, demonstrating that the two genes are key targets of Snail in neuroblasts. Mutation of the dCtBP co-repressor interaction motifs in the Snail protein leads to reduction of the Snail function in central nervous system. These results suggest that the Snail family of proteins control both asymmetry and cell division of neuroblasts by activating, probably indirectly, the expression of inscuteable and string. PMID- 11731457 TI - A role for Gsh1 in the developing striatum and olfactory bulb of Gsh2 mutant mice. AB - We have examined the role of the two closely related homeobox genes Gsh1 and Gsh2, in the development of the striatum and the olfactory bulb. These two genes are expressed in a partially overlapping pattern by ventricular zone progenitors of the ventral telencephalon. Gsh2 is expressed in both of the ganglionic eminences while Gsh1 is largely confined to the medial ganglionic eminence. Previous studies have shown that Gsh2(-/-) embryos suffer from an early misspecification of precursors in the lateral ganglionic eminence (LGE) leading to disruptions in striatal and olfactory bulb development. This molecular misspecification is present only in early precursor cells while at later stages the molecular identity of these cells appears to be normalized. Concomitant with this normalization, Gsh1 expression is notably expanded in the Gsh2(-/-) LGE. While no obvious defects in striatal or olfactory bulb development were detected in Gsh1(-/-) embryos, Gsh1/2 double homozygous mutants displayed more severe disruptions than were observed in the Gsh2 mutant alone. Accordingly, the molecular identity of LGE precursors in the double mutant is considerably more perturbed than in Gsh2 single mutants. These findings, therefore, demonstrate an important role for Gsh1 in the development of the striatum and olfactory bulb of Gsh2 mutant mice. In addition, our data indicate a role for Gsh genes in controlling the size of the LGE precursor pools, since decreasing copies of Gsh2 and Gsh1 alleles results in a notable decrease in precursor cell number, particularly in the subventricular zone. PMID- 11731458 TI - Functional equivalence of Hox gene products in the specification of the tritocerebrum during embryonic brain development of Drosophila. AB - Hox genes encode evolutionarily conserved transcription factors involved in the specification of segmental identity during embryonic development. This specification of identity is thought to be directed by differential Hox gene action, based on differential spatiotemporal expression patterns, protein sequence differences, interactions with co-factors and regulation of specific downstream genes. During embryonic development of the Drosophila brain, the Hox gene labial is required for the regionalized specification of the tritocerebral neuromere; in the absence of labial, the cells in this brain region do not acquire a neuronal identity and major axonal pathfinding deficits result. We have used genetic rescue experiments to investigate the functional equivalence of the Drosophila Hox gene products in the specification of the tritocerebral neuromere. Using the Gal4-UAS system, we first demonstrate that the labial mutant brain phenotype can be rescued by targeted expression of the Labial protein under the control of CNS-specific labial regulatory elements. We then show that under the control of these CNS-specific regulatory elements, all other Drosophila Hox gene products, except Abdominal-B, are able to efficiently replace Labial in the specification of the tritocerebral neuromere. We also observe a correlation between the rescue efficiency of the Hox proteins and the chromosomal arrangement of their encoding loci. Our results indicate that, despite considerably diverged sequences, most Hox proteins are functionally equivalent in their ability to replace Labial in the specification of neuronal identity. This suggests that in embryonic brain development, differences in Hox gene action rely mainly on cis acting regulatory elements and not on Hox protein specificity. PMID- 11731459 TI - Regionalisation of anterior neuroectoderm and its competence in responding to forebrain and midbrain inducing activities depend on mutual antagonism between OTX2 and GBX2. AB - The anterior neural ridge (ANR), and the isthmic organiser (IsO) represent two signalling centres possessing organising properties necessary for forebrain (ANR) as well as midbrain and rostral hindbrain (IsO) development. An important mediator of ANR and IsO organising property is the signalling molecule FGF8. Previous work has indicated that correct positioning of the IsO and Fgf8 expression in this domain is controlled by the transcription factors Otx2 and Gbx2. In order to provide novel insights into the roles of Otx2 and Gbx2, we have studied mutant embryos carrying different dosages of Otx2, Otx1 and Gbx2. Embryos deficient for both OTX2 and GBX2 proteins (hOtx1(2)/hOtx1(2); Gbx2(-/-)) show abnormal patterning of the anterior neural tissue, which is evident at the presomite-early somite stage prior to the onset of Fgf8 neuroectodermal expression. Indeed, hOtx1(2)/hOtx1(2); Gbx2(-/-) embryos exhibit broad co expression of early forebrain, midbrain and rostral hindbrain markers such as hOtx1, Gbx2, Pax2, En1 and Wnt1 and subsequently fail to activate forebrain and midbrain-specific gene expression. In this genetic context, Fgf8 is expressed throughout the entire anterior neural plate, thus indicating that its activation is independent of both OTX2 and GBX2 function. Analysis of hOtx1(2)/hOtx1(2); Gbx2(-/-) and Otx1(+/-); Otx2(+/-) mutant embryos also suggests that FGF8 cannot repress Otx2 without the participation of GBX2. Finally, we report that embryos carrying a single strong hypomorphic Otx2 allele (Otx2(lambda)) in an Otx2 and Gbx2 null background (Otx2(lambda)/-; Gbx2(-/-)) recover both the headless phenotype exhibited by Otx2(lambda)/- embryos and forebrain- and midbrain specific gene expression that is not observed in hOtx1(2)/hOtx1(2); Gbx2(-/-) mutants. Together, these data provide novel genetic evidence indicating that OTX2 and GBX2 are required for proper segregation of early regional identities anterior and posterior to the mid-hindbrain boundary (MHB) and for conferring competence to the anterior neuroectoderm in responding to forebrain-, midbrain- and rostral hindbrain-inducing activities. PMID- 11731460 TI - OTD/OTX2 functional equivalence depends on 5' and 3' UTR-mediated control of Otx2 mRNA for nucleo-cytoplasmic export and epiblast-restricted translation. AB - How gene activity is translated into phenotype and how it can modify morphogenetic pathways is of central importance when studying the evolution of regulatory control mechanisms. Previous studies in mouse have suggested that, despite the homeodomain-restricted homology, Drosophila orthodenticle (otd) and murine Otx1 genes share functional equivalence and that translation of Otx2 mRNA in epiblast and neuroectoderm might require a cell type-specific post transcriptional control depending on its 5' and 3' untranslated sequences (UTRs). In order to study whether OTD is functionally equivalent to OTX2 and whether synthesis of OTD in epiblast is molecularly dependent on the post-transcriptional control of Otx2 mRNA, we generated a first mouse model (otd(2)) in which an Otx2 region including 213 bp of the 5' UTR, exons, introns and the 3' UTR was replaced by an otd cDNA and a second mutant (otd(2FL)) replacing only exons and introns of Otx2 with the otd coding sequence fused to intact 5' and 3' UTRs of Otx2. otd(2) and otd(2FL) mRNAs were properly transcribed under the Otx2 transcriptional control, but mRNA translation in epiblast and neuroectoderm occurred only in otd(2FL) mutants. Phenotypic analysis revealed that visceral endoderm (VE) restricted translation of otd(2) mRNA was sufficient to rescue Otx2 requirement for early anterior patterning and proper gastrulation but it failed to maintain forebrain and midbrain identity. Importantly, epiblast and neuroectoderm translation of otd(2FL) mRNA rescued maintenance of anterior patterning as it did in a third mouse model replacing, as in otd(2FL), exons and introns of Otx2 with an Otx2 cDNA (Otx2(2c)). The molecular analysis has revealed that Otx2 5' and 3' UTR sequences, deleted in the otd(2) mRNA, are required for nucleo-cytoplasmic export and epiblast-restricted translation. Indeed, these molecular impairments were completely rescued in otd(2FL) and Otx2(2c) mutants. These data provide novel in vivo evidence supporting the concept that during evolution pre-existing gene functions have been recruited into new developmental pathways by modifying their regulatory control. PMID- 11731461 TI - Selective rescue of early haematopoietic progenitors in Scl(-/-) mice by expressing Scl under the control of a stem cell enhancer. AB - The stem cell leukaemia gene (Scl) encodes a basic helix-loop-helix transcription factor with a pivotal role in both haematopoiesis and endothelial development. During mouse development, Scl is first expressed in extra-embryonic mesoderm, and is required for the generation of all haematopoietic lineages and normal yolk sac angiogenesis. Ectopic expression of Scl during zebrafish development specifies haemangioblast formation from early mesoderm. These results suggest that SCL is essential for establishing the transcriptional programme responsible for the formation of haematopoietic stem cells and have focused attention on the transcriptional regulation of Scl itself. Previous studies have identified a panel of Scl enhancers each of which directed expression to a subdomain of the normal Scl expression pattern. Among them, a 3' enhancer directed expression during development to vascular endothelium and haematopoietic progenitors but not to Ter119(+) erythroid cells. The expression in haematopoietic stem cells, however, remained undetermined. We demonstrate that this 3' enhancer directs lacZ expression in transgenic mice to most foetal and adult long-term repopulating haematopoietic stem cells, and therefore functions as a stem cell enhancer. Consistent with these results, expression in Scl(-/-) embryos of exogenous Scl driven by the stem cell enhancer rescued the formation of early haematopoietic progenitors and also resulted in normal yolk sac angiogenesis. By contrast, erythropoiesis remained markedly deficient in rescued embryos. This observation is consistent with the inactivity of the stem cell enhancer in erythroid cells and reveals an essential role for SCL during erythroid differentiation in vivo. PMID- 11731462 TI - Transducing properties of Drosophila Frizzled proteins. AB - In Drosophila, two closely related serpentine receptors, Frizzled (Fz) and D Frizzled2 (Fz2) are able to act as receptors for the secreted Wnt peptide, Wingless (Wg). In addition to transducing the Wg signal, Fz (but not Fz2) is able to transduce a second, unidentified signal that mediates planar polarity. Much attention has been focused on the structure of the N-termini of the Fz-class receptors and their role in ligand binding. Experiments using techniques of high level expression have suggested a role for the C-termini in specifying which of the two second messenger systems the receptors are able to activate (M. Boutros, J. Mihaly, T. Bouwmeeste and M. Mlodzik (2000). Science 288, 1825-1828). We argue here that experiments involving high level expression of the receptors cannot be adequately interpreted and we have tested the ability of the receptors and chimeric forms when driven at moderate levels to rescue loss of function of the fz and fz2 genes. Under these conditions we find that all receptors tested will function as Wg receptors, but only a subset show the ability to rescue the polarity pathway. The presence of this subset implies that the N terminus is necessary but not sufficient and suggests that the ability to transduce the polarity signal is widely distributed throughout the protein. PMID- 11731463 TI - The functions of pannier during Drosophila embryogenesis. AB - The pannier (pnr) gene of Drosophila encodes a zinc-finger transcription factor of the GATA family and is involved in several developmental processes during embryonic and imaginal development. We report some novel aspects of the regulation and function of pnr during embryogenesis. Previous work has shown that pnr is activated by decapentaplegic (dpp) in early development, but we find that after stage 10, the roles are reversed and pnr becomes an upstream regulator of dpp. This function of pnr is necessary for the activation of the Dpp pathway in the epidermal cells implicated in dorsal closure and is not mediated by the JNK pathway, which is also necessary for Dpp activity in these cells. In addition, we show that pnr behaves as a selector-like gene in generating morphological diversity in the dorsoventral body axis. It is responsible for maintaining a subdivision of the dorsal half of the embryo into two distinct, dorsomedial and dorsolateral, regions, and also specifies the identity of the dorsomedial region. These results, together with prior work on its function in adults, suggest that pnr is a major factor in the genetic subdivision of the body of Drosophila. PMID- 11731464 TI - Mutations in LIKE HETEROCHROMATIN PROTEIN 1 affect flowering time and plant architecture in Arabidopsis. AB - In plants, recent studies have demonstrated links between the regulation of developmental processes and chromatin dynamics and organisation. Analysis of new mutations affecting overall plant architecture, leaf development and flowering time in Arabidopsis has allowed us to clone and characterise LHP1, the Drosophila heterochromatin protein 1 (HP1) homologue. LHP1 has the chromo and chromo shadow domains central to the function of animal proteins. Yeast two hybrid studies and in planta deletion experiments suggest similar modes of action in plants and animals via homodimer formation. In vivo localisation experiments revealed a specific subnuclear protein distribution in foci throughout the nucleus. Our data suggest that LHP1 may act as a main regulator of gene expression in plants, through formation of heterochromatin-like repressive complexes, to control developmental pathways involved in organ and cell size, and the vegetative to reproductive phase transition. PMID- 11731465 TI - Commissural axon pathfinding on the contralateral side of the floor plate: a role for B-class ephrins in specifying the dorsoventral position of longitudinally projecting commissural axons. AB - In both invertebrate and lower vertebrate species, decussated commissural axons travel away from the midline and assume positions within distinct longitudinal tracts. We demonstrate that in the developing chick and mouse spinal cord, most dorsally situated commissural neuron populations extend axons across the ventral midline and through the ventral white matter along an arcuate trajectory on the contralateral side of the floor plate. Within the dorsal (chick) and intermediate (mouse) marginal zone, commissural axons turn at a conserved boundary of transmembrane ephrin expression, adjacent to which they form a discrete ascending fiber tract. In vitro perturbation of endogenous EphB-ephrinB interactions results in the failure of commissural axons to turn at the appropriate dorsoventral position on the contralateral side of the spinal cord; consequently, axons inappropriately invade more dorsal regions of B-class ephrin expression in the dorsal spinal cord. Taken together, these observations suggest that B-class ephrins act locally during a late phase of commissural axon pathfinding to specify the dorsoventral position at which decussated commissural axons turn into the longitudinal axis. PMID- 11731466 TI - Fgf/MAPK signalling is a crucial positional cue in somite boundary formation. AB - The temporal and spatial regulation of somitogenesis requires a molecular oscillator, the segmentation clock. Through Notch signalling, the oscillation in cells is coordinated and translated into a cyclic wave of expression of hairy related and other genes. The wave sweeps caudorostrally through the presomitic mesoderm (PSM) and finally arrests at the future segmentation point in the anterior PSM. By experimental manipulation and analyses in zebrafish somitogenesis mutants, we have found a novel component involved in this process. We report that the level of Fgf/MAPK activation (highest in the posterior PSM) serves as a positional cue within the PSM that regulates progression of the cyclic wave and thereby governs the positions of somite boundary formation. PMID- 11731467 TI - Essential role of oligodendrocytes in the formation and maintenance of central nervous system nodal regions. AB - The membrane of myelinated axons is divided into functionally distinct domains characterized by the enrichment of specific proteins. The mechanisms responsible for this organization have not been fully identified. To further address the role of oligodendrocytes in the functional segmentation of the axolemma in vivo, the distribution of nodal (Na(+) channels, ankyrin G), paranodal (paranodin/contactin associated-protein) and juxtaparanodal (Kv1.1 K(+) channels) axonal markers, was studied in the brain of MBP-TK and jimpy mice. In MBP-TK transgenic mice, oligodendrocyte ablation was selectively induced by FIAU treatment before and during the onset of myelination. In jimpy mice, oligodendrocytes degenerate spontaneously within the first postnatal weeks after the onset of myelination. Interestingly, in MBP-TK mice treated for 1-20 days with FIAU, despite the ablation of more than 95% of oligodendrocytes, the protein levels of all tested nodal markers was unaltered. Nevertheless, these proteins failed to cluster in the nodal regions. By contrast, in jimpy mice, despite a diffused localization of paranodin, the formation of nodal clusters of Na(+) channels and ankyrin G was observed. Furthermore, K(+) channels clusters were transiently visible, but were in direct contact with nodal markers. These results demonstrate that the organization of functional domains in myelinated axons is oligodendrocyte dependent. They also show that the presence of these cells is a requirement for the maintenance of nodal and paranodal regions. PMID- 11731468 TI - Apoptosis--a death-inducing mechanism tightly linked with morphogenesis in Hydractina echinata (Cnidaria, Hydrozoa). AB - Programmed cell death is not only known as a mechanism mediating tissue destruction, but also as an organismic tool for body shaping and regulation of morphological events during development. Here we report the tight and vital link of the most prominent form of programmed cell death, apoptosis, to one of the oldest, most basic, and most radical developmental processes, the metamorphosis of the marine hydrozoon Hydractinia echinata. Apoptosis, represented by DNA fragmentation, appears very early during metamorphosis, approximately 20 minutes post induction. It is then executed in a very distinct spatial and temporal pattern, including the removal or phagocytosis of a large number of larval cells prior to the appearance of stolons and tentacles. Our data indicate a developmental program striving to reduce all body parts that are no longer necessary, before reaching a distinct turning point, when the development of adult features is initiated. During these events, morphogenesis of basal and apical structures correlates with recycling of that particular larval region, indicated by the presence of apoptosis. Based on these data, the necessity of apoptosis for normal development of adult patterns is inferred and a fundamental association of apoptosis with developmental processes can be stated. PMID- 11731469 TI - Overlapping functions of lysosomal acid phosphatase (LAP) and tartrate-resistant acid phosphatase (Acp5) revealed by doubly deficient mice. AB - To date, two lysosomal acid phosphatases are known to be expressed in cells of the monocyte/phagocyte lineage: the ubiquitously expressed lysosomal acid phosphatase (LAP) and the tartrate-resistant acid phosphatase-type 5 (Acp5). Deficiency of either acid phosphatase results in relatively mild phenotypes, suggesting that these enzymes may be capable of mutual complementation. This prompted us to generate LAP/Acp5 doubly deficient mice. LAP/Acp5 doubly deficient mice are viable and fertile but display marked alterations in soft and mineralised tissues. They are characterised by a progressive hepatosplenomegaly, gait disturbances and exaggerated foreshortening of long bones. Histologically, these animals are distinguished by an excessive lysosomal storage in macrophages of the liver, spleen, bone marrow, kidney and by altered growth plates. Microscopic analyses showed an accumulation of osteopontin adjacent to actively resorbing osteoclasts of Acp5- and LAP/Acp5-deficient mice. In osteoclasts of phosphatase-deficient mice, vacuoles were frequently found which contained fine filamentous material. The vacuoles in Acp5- and LAP/Acp5 doubly-deficient osteoclasts also contained crystallite-like features, as well as osteopontin, suggesting that Acp5 is important for processing of this protein. This is further supported by biochemical analyses that demonstrate strongly reduced dephosphorylation of osteopontin incubated with LAP/Acp5-deficient bone extracts. Fibroblasts derived from LAP/Acp5 deficient embryos were still able to dephosphorylate mannose 6-phosphate residues of endocytosed arylsulfatase A. We conclude that for several substrates LAP and Acp5 can substitute for each other and that these acid phosphatases are essential for processing of non-collagenous proteins, including osteopontin, by osteoclasts. PMID- 11731470 TI - Role of C. elegans lin-40 MTA in vulval fate specification and morphogenesis. AB - Vulval differentiation in Caenorhabditis elegans involves several fundamental cellular events, including cell fusion, division and migration. We have characterized the role of the lin-40 (also known as egr-1) gene in these cellular processes. LIN-40 is homologous to the metastasis-associated factor 1 (MTA1) in mammals, which has been identified as a component of the nucleosome remodeling and histone deacetylation (NuRD) complex that functions as a transcriptional co repressor. We show here that lin-40 negatively regulates vulval fate specification at least partly by promoting cell fusion between the vulval precursor cells and the hypodermal syncytium at an early larval stage. This inhibitory function of lin-40 might be carried out by downregulating lin-39 Hox expression. We also show that lin-40 is specifically required for cell divisions along the transverse orientation during vulval morphogenesis. PMID- 11731471 TI - The Drosophila ribbon gene encodes a nuclear BTB domain protein that promotes epithelial migration and morphogenesis. AB - During development of the Drosophila tracheal (respiratory) system, the cell bodies and apical and basal surfaces of the tracheal epithelium normally move in concert as new branches bud and grow out to form tubes. We show that mutations in the Drosophila ribbon (rib) gene disrupt this coupling: the basal surface continues to extend towards its normal targets, but movement and morphogenesis of the tracheal cell bodies and apical surface is severely impaired, resulting in long basal membrane protrusions but little net movement or branch formation. rib mutant tracheal cells are still responsive to the Branchless fibroblast growth factor (FGF) that guides branch outgrowth, and they express apical membrane markers normally. This suggests that the defect lies either in transmission of the FGF signal from the basal surface to the rest of the cell or in the apical cell migration and tubulogenesis machinery. rib encodes a nuclear protein with a BTB/POZ domain and Pipsqueak DNA-binding motif. It is expressed in the developing tracheal system and other morphogenetically active epithelia, many of which are also affected in rib mutants. We propose that Rib is a key regulator of epithelial morphogenesis that promotes migration and morphogenesis of the tracheal cell bodies and apical surface and other morphogenetic movements. PMID- 11731472 TI - A new RING for SUMO: wrestling transcriptional responses into nuclear bodies with PIAS family E3 SUMO ligases. PMID- 11731473 TI - Hedgehog signaling in animal development: paradigms and principles. PMID- 11731474 TI - PIASy, a nuclear matrix-associated SUMO E3 ligase, represses LEF1 activity by sequestration into nuclear bodies. AB - The Wnt-responsive transcription factor LEF1 can activate transcription in association with beta-catenin and repress transcription in association with Groucho. In search of additional regulatory mechanisms of LEF1 function, we identified the protein inhibitor of activated STAT, PIASy, as a novel interaction partner of LEF1. Coexpression of PIASy with LEF1 results in potent repression of LEF1 activity and in covalent modification of LEF1 with SUMO. PIASy markedly stimulates the sumoylation of LEF1 and multiple other proteins in vivo and functions as a SUMO E3 ligase for LEF1 in a reconstituted system in vitro. Moreover, PIASy binds to nuclear matrix-associated DNA sequences and targets LEF1 to nuclear bodies, suggesting that PIASy-mediated subnuclear sequestration accounts for the repression of LEF1 activity. PMID- 11731475 TI - Degradation of p53 by adenovirus E4orf6 and E1B55K proteins occurs via a novel mechanism involving a Cullin-containing complex. AB - Although MDM2 plays a major role in regulating the stability of the p53 tumor suppressor protein, other poorly understood MDM2-independent pathways also exist. Human adenoviruses have evolved strategies to regulate p53 function and stability to permit efficient viral replication. One mechanism involves adenovirus E1B55K and E4orf6 proteins, which collaborate to target p53 for degradation. To determine the mechanism of this process, a multiprotein E4orf6-associated complex was purified and shown to contain a novel Cullin-containing E3 ubiquitin ligase that is (1) composed of Cullin family member Cul5, Elongins B and C, and the RING H2 finger protein Rbx1(ROC1); (2) remarkably similar to the von Hippel-Lindau tumor suppressor and SCF (Skp1-Cul1/Cdc53-F-box) E3 ubiquitin ligase complexes; and (3) capable of stimulating ubiquitination of p53 in vitro in the presence of E1/E2 ubiquitin-activating and -conjugating enzymes. Cullins are activated by NEDD8 modification; therefore, to determine whether Cullin complexes are required for adenovirus-induced p53 degradation, studies were conducted in ts41 Chinese hamster ovary cells that are temperature sensitive for the NEDD8 pathway. E4orf6/E1B55K failed to induce the degradation of p53 at the nonpermissive temperature. Thus, our results identify a novel role for the Cullin-based machinery in regulation of p53. PMID- 11731476 TI - The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint. AB - The spindle checkpoint prevents cell cycle progression in cells that have mitotic spindle defects. Although several spindle defects activate the spindle checkpoint, the exact nature of the primary signal is unknown. We have found that the budding yeast member of the Aurora protein kinase family, Ipl1p, is required to maintain a subset of spindle checkpoint arrests. Ipl1p is required to maintain the spindle checkpoint that is induced by overexpression of the protein kinase Mps1. Inactivating Ipl1p allows cells overexpressing Mps1p to escape from mitosis and segregate their chromosomes normally. Therefore, the requirement for Ipl1p in the spindle checkpoint is not a consequence of kinetochore and/or spindle defects. The requirement for Ipl1p distinguishes two different activators of the spindle checkpoint: Ipl1p function is required for the delay triggered by chromosomes whose kinetochores are not under tension, but is not required for arrest induced by spindle depolymerization. Ipl1p localizes at or near kinetochores during mitosis, and we propose that Ipl1p is required to monitor tension at the kinetochore. PMID- 11731477 TI - c(3)G encodes a Drosophila synaptonemal complex protein. AB - The meiotic mutant c(3)G (crossover suppressor on 3 of Gowen) abolishes both synaptonemal complex (SC) formation and meiotic recombination, whereas mutations in the mei-W68 and mei-P22 genes prevent recombination but allow normal SC to form. These data, as well as a century of cytogenetic studies, support the argument that meiotic recombination between homologous chromosomes in Drosophila females requires synapsis and SC formation. We have cloned the c(3)G gene and shown that it encodes a protein that is structurally similar to SC proteins from yeast and mammals. Immunolocalization of the C(3)G protein, as well as the analysis of a C(3)G-eGFP expression construct, reveals that C(3)G is present in a thread-like pattern along the lengths of chromosomes in meiotic prophase, consistent with a role as an SC protein present on meiotic bivalents. The availability of a marker for SC in Drosophila allowed the investigation of the extent of synapsis in exchange-defective mutants. These studies indicate that SC formation is impaired in certain meiotic mutants and that the synaptic defect correlates with the exchange defects. Moreover, the observation of interference among the residual exchanges in these mutant oocytes implies that complete SC formation is not required for crossover interference in Drosophila. PMID- 11731478 TI - Histone H3 specific acetyltransferases are essential for cell cycle progression. AB - Longstanding observations suggest that acetylation and/or amino-terminal tail structure of histones H3 and H4 are critical for eukaryotic cells. For Saccharomyces cerevisiae, loss of a single H4-specific histone acetyltransferase (HAT), Esa1p, results in cell cycle defects and death. In contrast, although several yeast HAT complexes preferentially acetylate histone H3, the catalytic subunits of these complexes are not essential for viability. To resolve the apparent paradox between the significance of H3 versus H4 acetylation, we tested the hypothesis that H3 modification is essential, but is accomplished through combined activities of two enzymes. We observed that Sas3p and Gcn5p HAT complexes have overlapping patterns of acetylation. Simultaneous disruption of SAS3, the homolog of the MOZ leukemia gene, and GCN5, the hGCN5/PCAF homolog, is synthetically lethal due to loss of acetyltransferase activity. This key combination of activities is specific for these two HATs because neither is synthetically lethal with mutations of other MYST family or H3-specific acetyltransferases. Further, the combined loss of GCN5 and SAS3 functions results in an extensive, global loss of H3 acetylation and arrest in the G(2)/M phase of the cell cycle. The strikingly similar effect of loss of combined essential H3 HAT activities and the loss of a single essential H4 HAT underscores the fundamental biological significance of each of these chromatin-modifying activities. PMID- 11731479 TI - The yeast SAS (something about silencing) protein complex contains a MYST-type putative acetyltransferase and functions with chromatin assembly factor ASF1. AB - It is well established that acetylation of histone and nonhistone proteins is intimately linked to transcriptional activation. However, loss of acetyltransferase activity has also been shown to cause silencing defects, implicating acetylation in gene silencing. The something about silencing (Sas) 2 protein of Saccharomyces cerevisiae, a member of the MYST (MOZ, Ybf2/Sas3, Sas2, and TIP60) acetyltransferase family, promotes silencing at HML and telomeres. Here we identify a ~450-kD SAS complex containing Sas2p, Sas4p, and the tf2f related Sas5 protein. Mutations in the conserved acetyl-CoA binding motif of Sas2p are shown to disrupt the ability of Sas2p to mediate the silencing at HML and telomeres, providing evidence for an important role for the acetyltransferase activity of the SAS complex in silencing. Furthermore, the SAS complex is found to interact with chromatin assembly factor Asf1p, and asf1 mutants show silencing defects similar to mutants in the SAS complex. Thus, ASF1-dependent chromatin assembly may mediate the role of the SAS complex in silencing. PMID- 11731481 TI - Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways in Streptomyces coelicolor using DNA microarrays. AB - The eubacterial species Streptomyces coelicolor proceeds through a complex growth cycle in which morphological differentiation/development is associated with a transition from primary to secondary metabolism and the production of antibiotics. We used DNA microarrays and mutational analysis to investigate the expression of individual genes and multigene antibiotic biosynthetic pathways during these events. We identified expression patterns in biosynthetic, regulatory, and ribosomal protein genes that were associated highly specifically with particular stages of development. A knowledge-based algorithm that correlates temporal changes in expression with chromosomal position identified groups of contiguous genes expressed at discrete stages of morphological development, inferred the boundaries of known antibiotic synthesis gene loci, and revealed novel physical clusters of coordinately regulated genes. Microarray analysis of RNA from cells mutated in genes regulating synthesis of the antibiotics actinorhodin (Act) and undecylprodigiosin (Red) identified proximate and distant sites that contain putative ABC transporter and two-component system genes expressed coordinately with genes of specific biosynthetic pathways and indicated the existence of two functionally and physically discrete regulons in the Red pathway. PMID- 11731480 TI - The silencing complex SAS-I links histone acetylation to the assembly of repressed chromatin by CAF-I and Asf1 in Saccharomyces cerevisiae. AB - The acetylation state of histones plays a central role in determining gene expression in chromatin. The reestablishment of the acetylation state of nucleosomes after DNA replication and chromatin assembly requires both deacetylation and acetylation of specific lysine residues on newly incorporated histones. In this study, the MYST family acetyltransferase Sas2 was found to interact with Cac1, the largest subunit of Saccharomyces cerevisiae chromatin assembly factor-I (CAF-I), and with the nucleosome assembly factor Asf1. The deletions of CAC1 (cac1Delta), ASF1 (asf1Delta), and SAS2 (sas2Delta) had similar effects on gene silencing and were partially overlapping. Furthermore, Sas2 was found in a nuclear protein complex that included Sas4 and Sas5, a homolog of TAF(II)30. This complex, termed SAS-I, was also found to contribute to rDNA silencing. Furthermore, the observation that a mutation of H4 lysine 16 to arginine displayed the identical silencing phenotypes as sas2Delta suggested that it was the in vivo target of Sas2 acetylation. In summary, our data present a novel model for the reestablishment of acetylation patterns after DNA replication, by which SAS-I is recruited to freshly replicated DNA by its association with chromatin assembly complexes to acetylate lysine 16 of H4. PMID- 11731482 TI - Temporal regulation of a paired-like homeodomain repressor/TLE corepressor complex and a related activator is required for pituitary organogenesis. AB - Understanding the functional significance of the coordinate expression of specific corepressors and DNA-binding transcription factors remains a critical question in mammalian development. During the development of the pituitary gland, two highly related paired-like homeodomain factors, a repressor, Hesx1/Rpx and an activator, Prop-1, are expressed in sequential, overlapping temporal patterns. Here we show that while the repressive actions of Hesx1/Rpx may be required for initial pituitary organ commitment, progression beyond the appearance of the first pituitary (POMC) lineage requires both loss of Hesx1 expression and the actions of Prop-1. Although Hesx1 recruits both the Groucho-related corepressor TLE1 and the N-CoR/Sin3/HDAC complex on distinct domains, the repressor functions of Hesx1 in vivo prove to require the specific recruitment of TLE1, which exhibits a spatial and temporal pattern of coexpression during pituitary organogenesis. Furthermore, Hesx1-mediated repression coordinates a negative feedback loop with FGF8/FGF10 signaling in the ventral diencephalon, required to prevent induction of multiple pituitary glands from oral ectoderm. Our data suggest that the opposing actions of two structurally-related DNA-binding paired like homeodomain transcription factors, binding to similar cognate elements, coordinate pituitary organogenesis by reciprocally repressing and activating target genes in a temporally specific fashion, on the basis of the actions of a critical, coexpressed TLE corepressor. PMID- 11731483 TI - Cooperation between C/EBPalpha TBP/TFIIB and SWI/SNF recruiting domains is required for adipocyte differentiation. AB - Chromatin remodeling is an important step in promoter activation during cellular lineage commitment and differentiation. We show that the ability of the C/EBPalpha transcription factor to direct adipocyte differentiation of uncommitted fibroblast precursors and to activate SWI/SNF-dependent myeloid specific genes depends on a domain, C/EBPalpha transactivation element III (TE III), that binds the SWI/SNF chromatin remodeling complex. TE-III collaborates with C/EBPalpha TBP/TFIIB interaction motifs during induction of adipogenesis and adipocyte-specific gene expression. These results indicate that C/EBPalpha acts as a lineage-instructive transcription factor through SWI/SNF-dependent modification of the chromatin structure of lineage-specific genes, followed by direct promoter activation via recruitment of the basal transcription-initiation complex, and provide a mechanism by which C/EBPalpha can mediate differentiation along multiple cellular lineages. PMID- 11731484 TI - vhnf1, the MODY5 and familial GCKD-associated gene, regulates regional specification of the zebrafish gut, pronephros, and hindbrain. AB - Mutations in the homeobox gene vHnf1 are associated with human diseases MODY5 (maturity-onset diabetes of the young, type V) and familial GCKD (glomerulocystic kidney disease). In an insertional mutagenesis screen in zebrafish, we isolated mutant alleles of vhnf1. Phenotypes of these mutants include formation of kidney cysts, underdevelopment of the pancreas and the liver, and reduction in size of the otic vesicles. We show that these abnormalities arise from patterning defects during development. We further provide evidence that vhnf1 regulates the expression of key patterning genes for these organs. vhnf1 is required for the proper expression of pdx1 and shh (sonic hedgehog) in the gut endoderm, pax2 and wt1 in the pronephric primordial, and valentino (val) in the hindbrain. Complementary to the loss-of-function phenotypes, overexpression of vhnf1 induces expansion of the val expression domain in the hindbrain. We propose that vhnf1 controls development of multiple organs through regulating regional specification of organ primordia. The similarity between vhnf1-associated fish phenotypes and human symptoms suggests a correlation between developmental functions of vhnf1 and the molecular etiology of MODY5 and GCKD. PMID- 11731485 TI - Is there a bias in proteome research? AB - Advances in technology have enabled us to take a fresh look at data acquired by traditional single experiments and to compare them with genomewide data. The differences can be tremendous, as we show here, in the field of proteomics. We have compared data sets of protein-protein interactions in Saccharomyces cerevisiae that were detected by an identical underlying technical method, the yeast two-hybrid system. We found that the individually identified protein protein interactions are considerably different from those identified by two genomewide scans. Interacting proteins in the pooled database from single publications are much more closely related to each other with respect to transcription profiles when compared to genomewide data. This difference may have been introduced by two factors: by a selection process in individual publications and by false positives in the whole-genome scans. If we assume that the differences are a result of false positives in the whole-genome data, the scans would contain 47%, 44%, and 91% of false positives for the UETZ, ITO-core, and ITO-full data, respectively. If, however, the true fraction of false positives is considerably lower than estimated here, the data from hypothesis-driven experiments must have been subjected to a serious selection process. PMID- 11731486 TI - Most of the human genome is transcribed. PMID- 11731487 TI - A gene expression screen in zebrafish embryogenesis. AB - A screen for developmentally regulated genes was conducted in the zebrafish, a system offering substantial advantages for the study of the molecular genetics of vertebrate embryogenesis. Clones from a normalized cDNA library from early somitogenesis stages were picked randomly and tested by high-throughput in situ hybridization for restricted expression in at least one of four stages of development. Among 2765 clones that were screened, a total of 347 genes with patterns judged to be restricted were selected. These clones were subjected to partial sequence analysis, allowing recognition of functional motifs in 163 among them. In addition, a portion of the clones were mapped with the aid of the LN54 radiation hybrid panel. The usefulness of the in situ hybridization screening approach is illustrated by describing several new markers for the characteristic structure in the fish embryo named the yolk syncytial layer, and for different regions of the developing brain. PMID- 11731488 TI - Genomic dissection of genotype x environment interactions conferring adaptation of cotton to arid conditions. AB - The interaction of genotype with environment is of primary importance in many aspects of genomic research and is a special priority in the study of major crops grown in a wide range of environments. Water deficit, the major factor limiting plant growth and crop productivity worldwide, is expected to increase with the spread of arid lands. In genetically equivalent cotton populations grown under well-watered and water-limited conditions (the latter is responsible for yield reduction of approximately 50% relative to well-watered conditions), productivity and quality were shown to be partly accounted for by different quantitative trait loci (QTLs), indicating that adaptation to both arid and favorable conditions can be combined in the same genotype. QTL mapping was also used to test the association between productivity and quality under water deficit with a suite of traits often found to differ between genotypes adapted to arid versus well watered conditions. In this study, only reduced plant osmotic potential was clearly implicated in improved cotton productivity under arid conditions. Genomic tools and approaches may expedite breeding of genotypes that respond favorably to specific environments, help test roles of additional physiological factors, and guide the isolation of genes that protect crop performance under arid conditions toward improved adaptation of crops to arid cultivation. PMID- 11731489 TI - Novel protein domains and repeats in Drosophila melanogaster: insights into structure, function, and evolution. AB - Sequence database searching methods such as BLAST, are invaluable for predicting molecular function on the basis of sequence similarities among single regions of proteins. Searches of whole databases however, are not optimized to detect multiple homologous regions within a single polypeptide. Here we have used the prospero algorithm to perform self-comparisons of all predicted Drosophila melanogaster gene products. Predicted repeats, and their homologs from all species, were analyzed further to detect hitherto unappreciated evolutionary relationships. Results included the identification of novel tandem repeats in the human X-linked retinitis pigmentosa type-2 gene product, repeated segments in cystinosin, associated with a defect in cystine transport, and 'nested' homologous domains in dysferlin, whose gene is mutated in limb girdle muscular dystrophy. Novel signaling domain families were found that may regulate the microtubule-based cytoskeleton and ubiquitin-mediated proteolysis, respectively. Two families of glycosyl hydrolases were shown to contain internal repetitions that hint at their evolution via a piecemeal, modular approach. In addition, three examples of fruit fly genes were detected with tandem exons that appear to have arisen via internal duplication. These findings demonstrate how completely sequenced genomes can be exploited to further understand the relationships between molecular structure, function, and evolution. PMID- 11731490 TI - Evolution of gene order in the genomes of two related yeast species. AB - Changes in gene order between the genomes of two related yeast species, Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum were studied. From the dataset of a previous low coverage sequencing of the S. bayanus var. uvarum genome, 35 different synteny breakpoints between neighboring genes and two cases of local gene inversion were characterized in detail. The number and the type of the chromosomal rearrangements that have led to these differences were identified. We show that evolution of gene order in the genomes of these two yeast species is driven mainly by gene duplication onto different chromosomes followed by differential loss of the repeated copies. In addition, local gene inversions also would result from a mechanism of gene duplication, but in an inverted orientation, followed by loss of the original copy. The identification of traces of anciently duplicated genes, called relics, show that the loss of duplicates is more frequently caused by the accumulation of numerous mutations in one of the two copies than by DNA deletion. Surprisingly, gross chromosomal rearrangements such as translocations have only a minor effect on gene order reshuffling as they account for <10% of the synteny breakpoints. PMID- 11731491 TI - Comparative genomics between rice and Arabidopsis shows scant collinearity in gene order. AB - We have investigated possible collinearity between the genomes of rice and Arabidopsis by comparing 126 annotated and mapped rice BAC sequences (approximately 20 Mb of sequence) with the annotated and complete Arabidopsis genome (approximately 115 Mb). Although we were able to identify several regions in which gene order is preserved, they are relatively small, and are interrupted by noncollinear genes. Computer simulation showed that these microscale collinearities are above the expectation for a random process. On the other hand, the order of exons within homologous genes (<2.5 kb) was preserved, as expected. PMID- 11731492 TI - Identification of wound healing/regeneration quantitative trait loci (QTL) at multiple time points that explain seventy percent of variance in (MRL/MpJ and SJL/J) mice F2 population. AB - Studies on genetic mechanisms of wound healing in mammals are very few, although injury is a leading cause of the global burden of disease. In this study, we performed a high-density, genome-wide scan using 633 (MRL/MPJ x SJL/J) F(2) intercross at multiple time points (days 15, 21, and 25) to identify quantitative trait loci (QTL) involved in wound healing/regeneration. The hypothesis of the study was that QTL and unique epistatic interactions are involved at each time point to promote wound healing/regeneration. Ten QTL were identified from chromosomes 1, 4, 6, 7, 9, and 13. Of the 10 QTL, eight from chromosomes 1, 4, 6, and 9 were novel as compared to QTL identified in the study. The 10 QTL altogether explained 70% of variance in F(2) mice. The same QTL were identified at each time point, with simple linear correlation between days 15, 21, and 25, showing very high significant relationships (R >0.92, P <0.0001). Unique epistatic interactions were identified at each time point except those from chromosomes 4, 6, 9, and 13 that were found at all three time points, showing that some loci are involved at all the three time points of wound healing (days 15, 21, and 25). Therefore, loci-to-loci interactions may play a major role in wound healing. Information from these studies may help in the identification of genes that could be involved in wound healing/regeneration. PMID- 11731493 TI - Identification of genes encoding amino acid permeases by inactivation of selected ORFs from the Synechocystis genomic sequence. AB - Genes encoding elements of four amino acid permeases were identified by insertional inactivation of ORFs from the genomic sequence of the cyanobacterium Synechocystis sp. strain PCC 6803 whose putative products are homologous to amino acid permease proteins from other bacteria. A transport system for neutral amino acids and histidine and a transport system for basic amino acids and glutamine were identified as ABC-type transporters, whereas Na(+)-dependent transport of glutamate was found to be mediated by at least two systems, the secondary permease GltS and a TRAP-type transporter. Except for GltS, substrate specificities of the identified permeases do not match those of previously characterized systems homologous to these permeases. PMID- 11731494 TI - Envelope-class retrovirus-like elements are widespread, transcribed and spliced, and insertionally polymorphic in plants. AB - Retrotransposons and retroviruses share similar intracellular life cycles and major encoded proteins, but retrotransposons lack the envelope (env) critical for infectivity. Retrotransposons are ubiquitous and abundant in plants and active retroviruses are known in animals. Although a few env-containing retroelements, gypsy-like Athila, Cyclops, and Calypso and copia-like SIRE-1, have been identified in plants, the general presence and functionality of the domain remains unclear. We show here that env-class elements are present throughout the flowering plants and are widely transcribed. Within the grasses, we show the transcription of the env domain itself for Bagy-2 and related retrotransposons, all members of the Athila group. Furthermore, Bagy-2 transcripts undergo splicing to generate a subgenomic env product as do those of retroviruses. Transcription and the polymorphism of their insertion sites in closely related barley cultivars suggests that at least some are propagationally active. The putative ENV polypeptides of Bagy-2 and rice Rigy-2 contain predicted leucine zipper and transmembrane domains typical of retroviral ENVs. These findings raise the prospect of active retroviral agents among the plants. PMID- 11731495 TI - Genomic characterization of recent human LINE-1 insertions: evidence supporting random insertion. AB - LINE-1 (L1) elements play an important creative role in genomic evolution by distributing both L1 and non-L1 DNA in a process called retrotransposition. A large percentage of the human genome consists of DNA that has been dispersed by the L1 transposition machinery. L1 elements are not randomly distributed in genomic DNA but are concentrated in regions with lower GC content. In an effort to understand the consequences of L1 insertions, we have begun an investigation of their genomic characteristics and the changes that occur to them over time. We compare human L1 insertions that were created either during recent human evolution or during the primate radiation. We report that L1 insertions are an important source for the creation of new microsatellites. We provide evidence that L1 first strand cDNA synthesis can occur from an internal priming event. We note that in contrast to older L1 insertions, recent L1s are distributed randomly in genomic DNA, and the shift in the L1 genomic distribution occurs relatively rapidly. Taken together, our data indicate that strong forces act on newly inserted L1 retrotransposons to alter their structure and distribution. PMID- 11731496 TI - Twin priming: a proposed mechanism for the creation of inversions in L1 retrotransposition. AB - L1 retrotransposons are pervasive in the human genome. Approximately 25% of recent L1 insertions in the genome are inverted and truncated at the 5' end of the element, but the mechanism of L1 inversion has been a complete mystery. We analyzed recent L1 inversions from the genomic database and discovered several findings that suggested a mechanism for the creation of L1 inversions, which we call twin priming. Twin priming is a consequence of target primed reverse transcription (TPRT), a coupled reverse transcription/integration reaction that L1 elements are thought to use during their retrotransposition. In TPRT, the L1 endonuclease cleaves DNA at its target site to produce a double-strand break with two single-strand overhangs. During twin priming, one of the overhangs anneals to the poly(A) tail of the L1 RNA, and the other overhang anneals internally on the RNA. The overhangs then serve as primers for reverse transcription. The data further indicate that a process identical to microhomology-driven single-strand annealing resolves L1 inversion intermediates. PMID- 11731497 TI - Evolutionary history of Cer elements and their impact on the C. elegans genome. AB - We report the results of sequence analysis and chromosomal distribution of all distinguishable long terminal repeat (LTR) retrotransposons (Cer elements) in the Caenorhabditis elegans genome. Included in this analysis are all readily recognizable full-length and fragmented elements, as well as solo LTRs. Our results indicate that there are 19 families of Cer elements, some of which display significant subfamily structure. Cer elements can be clustered based on their tRNA primer binding sites (PBSs). These clusters are in concordance with our reverse transcriptase- and LTR-based phylogenies. Although we find that most Cer elements are located in the gene depauperate chromosome ends, some elements are located in or near putative genes and may contribute to gene structure and function. The results of RT-PCR analyses are consistent with this prediction. PMID- 11731498 TI - QTL analysis in a complex autopolyploid: genetic control of sugar content in sugarcane. AB - QTL mapping in autopolyploids is complicated by the possibility of segregation for three or more alleles at a locus and by a lack of preferential pairing, however the subset of polymorphic alleles that show simplex segregation ratios can be used to locate QTLs. In autopolyploid Saccharum, 36 significant associations between variation in sugar content and unlinked loci detected by 31 different probes were found in two interspecific F(1) populations. Most QTL alleles showed phenotypic effects consistent with the parental phenotypes, but occasional transgressive QTLs revealed opportunities to purge unfavorable alleles from cultivars or introgress valuable alleles from exotics. Several QTLs on homologous chromosomes appeared to correspond to one another-multiple doses of favorable 'alleles' at such chromosomal region(s) yielded diminishing returns such negative epistasis may contribute to phenotypic buffering. Fewer sugar content QTLs were discovered from the highest-sugar genotype than from lower sugar genotypes, perhaps suggesting that many favorable alleles have been fixed by prior selection, i.e. that the genes for which allelic variants (QTLs) persist in improved sugarcanes may be a biased subset of the population of genes controlling sugar content. Comparison of these data to mutations and QTLs previously mapped in maize hinted that seed and biomass crops may share a partly overlapping basis for genetic variation in carbohydrate deposition. However, many QTLs do not correspond to known candidate genes, suggesting that other approaches will be necessary to isolate the genetic determinants of high sugar content of vegetative tissues. PMID- 11731499 TI - Comparative sequence analysis of the imprinted Dlk1-Gtl2 locus in three mammalian species reveals highly conserved genomic elements and refines comparison with the Igf2-H19 region. AB - The Dlk1-Gtl2 domain on mouse chromosome 12 contains reciprocally imprinted genes with the potential to contribute to our understanding of common features involved in imprinting control. We have sequenced this conserved region in the mouse and sheep and included the human sequence in a three species comparison. This analysis resulted in a precise conservation map and identification of highly conserved sequence elements, some of which we have shown previously to be differentially methylated in the mouse. Additionally, this analysis facilitated identification of a CpG-rich tandem repeat array located approximately 13-15 kb upstream of Gtl2. Furthermore, we have identified a third imprinted transcript that overlaps with the last Dlk1 exon in the mouse. This transcript lacks a conserved open reading frame and is probably generated by cleavage of extended Dlk1 transcripts. Because Dlk1 and Gtl2 share many of the imprinting properties of the well-characterized Igf2-H19 domain, it has been proposed that the two regions may be regulated in the same way. Comparative genomic examination of the two domains indicates that although there are similarities, other features are very different, including the location of conserved CTCF-binding sites, and the level of conservation at regulatory regions. PMID- 11731500 TI - Differential divergence of three human pseudoautosomal genes and their mouse homologs: implications for sex chromosome evolution. AB - The human pseudoautosomal region 1 (PAR1) is essential for meiotic pairing and recombination, and its deletion causes male sterility. Comparative studies of human and mouse pseudoautosomal genes are valuable in charting the evolution of this interesting region, but have been limited by the paucity of genes conserved between the two species. We have cloned a novel human PAR1 gene, DHRSXY, encoding an oxidoreductase of the short-chain dehydrogenase/reductase family, and isolated a mouse ortholog Dhrsxy. We also searched for mouse homologs of recently reported PGPL and TRAMP genes that flank it within PAR1. We recovered a highly conserved mouse ortholog of PGPL by cross-hybridization, but found no mouse homolog of TRAMP. Like Csf2ra and Il3ra, both mouse homologs are autosomal; Pgpl on chromosome 5, and Dhrsxy subtelomeric on chromosome 4. TRAMP, like the human genes within or near PAR1, is probably very divergent or absent in the mouse genome. We interpret the rapid divergence and loss of pseudoautosomal genes in terms of a model of selection for the concentration of repetitive recombinogenic sequences that predispose to high recombination and translocation. PMID- 11731501 TI - Evolution of intron/exon structure of DEAD helicase family genes in Arabidopsis, Caenorhabditis, and Drosophila. AB - The DEAD box RNA helicase (RH) proteins are homologs involved in diverse cellular functions in all of the organisms from prokaryotes to eukaryotes. Nevertheless, there is a lack of conservation in the splicing pattern in the 53 Arabidopsis thaliana (AtRHs), the 32 Caenorhabditis elegans (CeRHs) and the 29 Drosophila melanogaster (DmRHs) genes. Of the 153 different observed intron positions, 4 are conserved between AtRHs, CeRHs, and DmRHs, and one position is also found in RHs from yeast and human. Of the 27 different AtRH structures with introns, 20 have at least one predicted ancient intron in the regions coding for the catalytic domain. In all of the organisms examined, we found at least one gene with most of its intron predicted to be ancient. In A. thaliana, the large diversity in RH structures suggests that duplications of the ancestral RH were followed by a high number of intron deletions and additions. The very high bias toward phase 0 introns is in favor of intron addition, preferentially in phase 0. Results from this comparative study of the same gene family in a plant and in two animals are discussed in terms of the general mechanisms of gene family evolution. PMID- 11731502 TI - Trimming, weighting, and grouping SNPs in human case-control association studies. AB - The search for genes underlying complex traits has been difficult and often disappointing. The main reason for these difficulties is that several genes, each with rather small effect, might be interacting to produce the trait. Therefore, we must search the whole genome for a good chance to find these genes. Doing this with tens of thousands of SNP markers, however, greatly increases the overall probability of false-positive results, and current methods limiting such error probabilities to acceptable levels tend to reduce the power of detecting weak genes. Investigating large numbers of SNPs inevitably introduces errors (e.g., in genotyping), which will distort analysis results. Here we propose a simple strategy that circumvents many of these problems. We develop a set-association method to blend relevant sources of information such as allelic association and Hardy-Weinberg disequilibrium. Information is combined over multiple markers and genes in the genome, quality control is improved by trimming, and an appropriate testing strategy limits the overall false-positive rate. In contrast to other available methods, our method to detect association to sets of SNP markers in different genes in a real data application has shown remarkable success. PMID- 11731503 TI - Identification of potential interaction networks using sequence-based searches for conserved protein-protein interactions or "interologs". AB - Protein interaction maps have provided insight into the relationships among the predicted proteins of model organisms for which a genome sequence is available. These maps have been useful in generating potential interaction networks, which have confirmed the existence of known complexes and pathways and have suggested the existence of new complexes and or crosstalk between previously unlinked pathways. However, the generation of such maps is costly and labor intensive. Here, we investigate the extent to which a protein interaction map generated in one species can be used to predict interactions in another species. PMID- 11731504 TI - The LN54 radiation hybrid map of zebrafish expressed sequences. AB - To increase the density of a gene map of the zebrafish, Danio rerio, we have placed 3119 expressed sequence tags (ESTs) and cDNA sequences on the LN54 radiation hybrid (RH) panel. The ESTs and genes mapped here join 748 SSLp markers and 459 previously mapped genes and ESTs, bringing the total number of markers on the LN54 RH panel to 4226. Addition of these new markers brings the total LN54 map size to 14,372 cR, with 118 kb/cR. The distribution of ESTs according to linkage groups shows relatively little variation (minimum, 73; maximum, 201). This observation, combined with a relatively uniform size for zebrafish chromosomes, as previously indicated by karyotyping, indicates that there are no especially gene-rich or gene-poor chromosomes in this species. We developed an algorithm to provide a semiautomatic method for the selection of additional framework markers for the LN54 map. This algorithm increased the total number of framework markers to 1150 and permitted the mapping of a high percentage of sequences that could not be placed on a previous version of the LN54 map. The increased concentration of expressed sequences on the LN54 map of the zebrafish genome will facilitate the molecular characterization of mutations in this species. PMID- 11731505 TI - Toward a cytological characterization of the rice genome. AB - Rice (Oryza sativa L.) will be the first major crop, as well as the first monocot plant species, to be completely sequenced. Integration of DNA sequence-based maps with cytological maps will be essential to fully characterize the rice genome. We have isolated a set of 24 chromosomal arm-specific bacterial artificial chromosomes to facilitate rice chromosome identification. A standardized rice karyotype was constructed using meiotic pachytene chromosomes of O. sativa spp. japonica rice var. Nipponbare. This karyotype is anchored by centromere-specific and chromosomal arm-specific cytological landmarks and is fully integrated with the most saturated rice genetic linkage maps in which Nipponbare was used as one of the mapping parents. An ideogram depicting the distribution of heterochromatin in the rice genome was developed based on the patterns of 4',6-diamidino-2 phenylindole staining of the Nipponbare pachytene chromosomes. The majority of the heterochromatin is distributed in the pericentric regions with some rice chromosomes containing a significantly higher proportion of heterochromatin than other chromosomes. We showed that pachytene chromosome-based fluorescence in situ hybridization analysis is the most effective approach to integrate DNA sequences with euchromatic and heterochromatic features. PMID- 11731507 TI - GeneLynx: a gene-centric portal to the human genome. AB - GeneLynx is a meta-database providing an extensive collection of hyperlinks to human gene-specific information in diverse databases available on the Internet. The GeneLynx project is based on the simple notion that given any gene-specific identifier (accession number, gene name, text, or sequence), scientists should be able to access a single location that provides a set of links to all the publicly available information pertinent to the specified human gene. GeneLynx was implemented as an extensible relational database with an intuitive and user friendly Web interface. The data are automatically extracted from more than 40 external resources, using appropriate approaches to maximize coverage of the available data. Construction and curation of the system is mediated by a custom set of software tools. An indexing utility is provided to facilitate the establishment of hyperlinks in external databases. A unique feature of the GeneLynx system is a communal curation system for user-aided annotation. GeneLynx can be accessed freely at http://www.genelynx.org. PMID- 11731508 TI - Are we the barrier? PMID- 11731509 TI - Lack of adherence with the analgesic regimen: a significant barrier to effective cancer pain management. AB - PURPOSE: To evaluate oncology outpatients' level of adherence to their analgesic regimen during a 5-week period. PATIENTS AND METHODS: A random sample of 65 adult oncology outpatients with a Karnofsky performance status score of >or= 50, an average pain intensity score of >or= 2.5, and radiographic evidence of bone metastasis were recruited for this longitudinal study from seven outpatient settings. On a daily basis, patients rated their level of pain intensity and recorded pain medication intake. Adherence rates for opioid analgesics prescribed on an around-the-clock (ATC) and on an as-needed (PRN) basis were calculated on a weekly basis. RESULTS: Overall adherence rates for ATC opioid analgesics ranged from 84.5% to 90.8% and, for PRN analgesics, from 22.2% to 26.6%. No significant differences over time were found in either of these adherence rates. CONCLUSION: One factor that seems to contribute to ineffective cancer pain management is poor adherence to the analgesic regimen. PMID- 11731510 TI - Methodologic lessons learned from hot flash studies. AB - PURPOSE: In the course of conducting a series of prospective clinical trials devoted to defining new treatment opportunities for hot flashes in cancer survivors, considerable experience has been acquired with related methodologic issues. This article has been written in response to many queries regarding this methodology. PATIENTS AND METHODS: A series of seven different clinical trials that involved 968 patients was used for this work. Reliable and valid definitions of hot flash intensity were developed from patient-reported descriptions. Concomitant validity and reliability assessment of patient-completed diaries was undertaken to compare hot flash data with toxicity and quality-of-life (QOL) end points and to examine consistency across patient groups using variability analysis and correlation procedures. Parametric data from this meta-analysis was used to examine relative power considerations for the design of phase II and phase III clinical trials. RESULTS: Daily diaries used in these studies exhibited consistency and reliability and had few missing data. Hot flash frequency and hot flash score (frequency multiplied by average severity) variables produced almost identical end point results. For phase III placebo-controlled studies, 50 patients per treatment arm seem appropriate to provide sufficient power specifications to detect a clinically meaningful change in hot flash activity. For phase II trials, 25 patients per trial seem to provide reasonable estimates of eventual hot flash efficacy to screen potential agents for more definitive testing. CONCLUSION: Given the data gained from these experiences, we can plan and carry out more efficient trials to identify efficacious agents for the reduction of hot flash activity. PMID- 11731506 TI - Advanced integrated mouse YAC map including BAC framework. AB - Functional characterization of the mouse genome requires the availability of a comprehensive physical map to obtain molecular access to chromosomal regions of interest. Positional cloning remains a crucial way of linking phenotype with particular genes. A key step and frequent stumbling block in positional cloning is making a contig of a genetically defined candidate region. The most efficient first step is isolating YAC (Yeast Artificial Chromosome) clones. A robust, detailed YAC contig map is thus an important tool. Employing Interspersed Repetitive Sequence (IRS)-PCR genomics, we have generated an advanced second generation YAC contig map of the mouse genome that doubles both the depth of clones and the density of markers available. In addition to the primarily YAC based map, we located 1942 BAC (Bacterial Artificial Chromosome) clones. This allows us to present for the first time a dense framework of BACs spanning the genome of the mouse, which, for instance, can serve as a nucleus for genomic sequencing. Four large-insert mouse YAC libraries from three different strains are included in our data, and our analysis incorporates the data of Hunter et al. and Nusbaum et al. There is a total of 20,205 markers on the final map, 12,033 from our own data, and a total of 56,093 YACs, of which 44,401 are positive for more than one marker. PMID- 11731511 TI - Impact of the media and the internet on oncology: survey of cancer patients and oncologists in Canada. AB - PURPOSE: To evaluate the use of the news media and the Internet as sources of medical information by patients and oncologists in Canada and to investigate the impact on patients' treatment decisions and the patient-doctor relationship. PATIENTS AND METHODS: During a 2-week period, 191 ambulatory patients participated in the survey. Questionnaires were also mailed to Canadian oncologists: 410 of 686 questionnaires were returned (response rate = 60%). RESULTS: Of the 191 patients, 86% wanted as much information as possible about their illness, 54% reported receiving insufficient information, 83% cited physicians as their primary information source, and 7% cited the Internet. Seventy-one percent of patients actively searched for information, and 50% used the Internet. Patients' opinions about the balance, accuracy, and relevance of news media reports were evenly split. English as the first language, access to the Internet, and use of alternative treatments predicted a higher rate of information seeking. Most oncologists routinely pay some attention to medical news and believe that it is difficult for patients to interpret medical information in the media and on the Internet accurately. Both patients and oncologists agree that information seeking does not affect the patient-physician relationship. CONCLUSION: Information searching is common among cancer patients in Canada. It does not affect the patient-doctor relationship. The media and the Internet are powerful means of medical information dissemination. Strategic efforts are needed to improve the quality of medical news reporting by the media, and to provide guidance for patients to understand their disease and interpret such information better. PMID- 11731512 TI - ERCC1 and thymidylate synthase mRNA levels predict survival for colorectal cancer patients receiving combination oxaliplatin and fluorouracil chemotherapy. AB - PURPOSE: To test the hypotheses of whether the relative mRNA expression of the thymidylate synthase (TS) gene and the excision cross-complementing (ERCC1) gene are associated with response to and survival of fluorouracil (5-FU)/oxaliplatin chemotherapy in metastatic colorectal cancer. PATIENTS AND METHODS: Patients had progressive stage IV disease after unsuccessful 5-FU and irinotecan chemotherapy. All patients were evaluated for eligibility for a compassionate 5-FU/oxaliplatin protocol. cDNA was derived from paraffin-embedded tumor specimens to determine TS and ERCC1 mRNA expression relative to the internal reference gene beta-actin using fluorescence-based, real-time reverse transcriptase polymerase chain reaction. RESULTS: The median TS gene expression level from 50 metastasized tumors was 3.4 x 10(-3) (minimum expression, 0.18 x 10(-3);maximum expression, 11.5 x 10(-3)), and the median ERCC1 gene expression level was 2.53 x 10(-3) (minimum, 0.0; maximum, 14.61 x 10(-3)). The gene expression cutoff values for chemotherapy nonresponse were 7.5 x 10(-3) for TS and 4.9 x 10(-3) for ERCC1. The median survival time for patients with TS 6 cm), or elevated serum lactate dehydrogenase (LDH) level. Patients were given three cycles of chemotherapy before radiotherapy. The chemotherapy comprised a 3-week cycle of mitomycin, epirubicin, and cisplatin on day 1 and fluorouracil and leucovorin on day 8 (MEPFL). RESULTS: From January 1994 to December 1997, 111 patients were recruited. The median follow-up period was 43 months. The actuarial 5-year overall survival rate was 70% (95% confidence interval [CI], 60% to 80%; n = 111). For patients having completed radiotherapy (n = 100), the 5-year locoregional control rate was 70% (95% CI, 55% to 84%) and the distant metastasis-free rate was 81% (95% CI, 73% to 89%). The 5-year distant metastasis-free rate of N3a and N3b disease of AJCC 1997 staging system were 79% (95% CI, 62% to 95%) and 74% (95% CI, 60% to 89%), respectively. By Cox multivariate analysis, high pretreatment serum LDH level (P = .04) and neck nodal enlargement before radiotherapy (P = .001) were adverse prognostic factors of survival. CONCLUSION: The good 5-year survival of N3 disease supports the effectiveness of induction MEPFL in the primary treatment of advanced NPC. Further investigation to incorporate concurrent chemoradiotherapy is warranted. PMID- 11731514 TI - Long-term results of blood and marrow transplantation for Hodgkin's lymphoma. AB - PURPOSE: To evaluate the long-term outcome after allogeneic (allo) and autologous (auto) blood or marrow transplantation (BMT) in patients with relapsed or refractory Hodgkin's lymphoma (HL). PATIENTS AND METHODS: We analyzed the outcome of 157 consecutive patients with relapsed or refractory HL, who underwent BMT between March 1985 and April 1998. Patients 93%) in beta3-/- granule cells, the local inhibitory interneurons of the OB. This was revealed by a large reduction of muscimol-evoked whole-cell current and the total current mediated by spontaneous, miniature inhibitory postsynaptic currents (mIPSCs). In beta3-/- mitral/tufted cells (principal cells), there was a two-fold increase in mIPSC amplitudes without any significant change in their kinetics or frequency. In parallel with the altered inhibition, there was a significant increase in the amplitude of theta (80% increase) and gamma (178% increase) frequency oscillations in beta3-/- OBs recorded in vivo from freely moving mice. In odor discrimination tests, we found beta3-/- mice to be initially the same as, but better with experience than beta3+/+ mice in distinguishing closely related monomolecular alcohols. However, beta3-/- mice were initially better and then worse with practice than control mice in distinguishing closely related mixtures of alcohols. Our results indicate that the disruption of GABA(A) receptor mediated synaptic inhibition of GABAergic interneurons and the augmentation of IPSCs in principal cells result in increased network oscillations in the OB with complex effects on olfactory discrimination, which can be explained by an increase in the size or effective power of oscillating neural cell assemblies among the mitral cells of beta3-/- mice. PMID- 11731540 TI - Putative feed-forward control of jaw-closing muscle activity during rhythmic jaw movements in the anesthetized rabbit. AB - When a thin plastic test strip of various hardness is placed between the upper and lower teeth during rhythmical jaw movements induced by electrical stimulation of the cortical masticatory area (CMA) in anesthetized rabbits, electromyographic (EMG) activity of the masseter muscle is facilitated in a hardness-dependent manner. This facilitatory masseteric response (FMR) often occurred prior to contact of the teeth to the strip, and thus preceded the onset of the masticatory force. Since this finding suggests involvement of a feed-forward mechanism in the induction of the FMR, the temporal relationship between the onset of the FMR and that of the masticatory force was analyzed in five sequential masticatory cycles after application of the strip. The FMR was found to precede the onset of masticatory force from the second masticatory cycle after application of the strip, but never did in the first cycle. This finding supports the concept of a feed-forward control mechanism that modulates FMR timing. Furthermore, the FMR preceding the force onset disappeared after making a lesion of the mesencephalic trigeminal nucleus (MesV) where the ganglion cells of the muscle spindle afferents from the jaw-closing muscles are located. In contrast, no such change occurred after blocking periodontal afferents by transection of both the maxillary and the inferior alveolar nerves. The putative feed-forward control of the FMR is therefore dependent mainly on sensory inputs from the muscle spindles, but little on those from the periodontal receptors, if any. We further examined the involvement of the CMA with the putative feed-forward control of the FMR via the transcortical loop. For this purpose, rhythmical jaw movements were induced by stimulation of the pyramidal tract. No significant change in the timing of the FMR occurred after the CMA ablation, which strongly suggests that the CMA is not involved in the putative feed-forward control of the FMR. The FMR was also noted to increase significantly in a hardness-dependent manner even after the MesV lesion, although the rate of increment decreased significantly. Contribution of muscle spindles and periodontal receptors to the hardness-dependent change of the FMR is discussed. PMID- 11731541 TI - Evidence that post-tetanic potentiation is mediated by neuropeptide release in Aplysia. AB - Many neuromuscular and central synapses exhibit activity-dependent plasticity. The sustained high-frequency firing needed to elicit some forms of plasticity are similar to those often required to release neuropeptides. We wanted to determine if neuropeptide release could contribute to post-tetanic potentiation (PTP) and chose neuromuscular synapses in buccal muscle I3a to explore this issue. This muscle is innervated by two motor neurons (termed B3 and B38) that show PTP in response to tetanic stimulation. B3 and B38 use glutamate as their fast transmitter but express different modulatory neuropeptides. B3 expresses FMRFamide, a neuropeptide that only slightly increases its own excitatory junction potentials (EJPs). B38 expresses the small cardioactive peptide (SCP), a neuropeptide that dramatically increases its own EJPs. It was our hypothesis that SCP released from B38's terminals during tetanic stimulation mediated a component of PTP for B38. Because no antagonist to SCP currently exists, we used several indirect approaches to test this hypothesis. First, we studied the effects of increasing stimulation frequency during the tetanus or lowering temperature on PTP. Both of these changes are known to dramatically increase SCP release. We found that increasing the frequency of stimulation increased PTP for both neurons; however, the effects were larger for B38. Decreasing the temperature tended to reduce PTP for B3, while increasing PTP for B38. These results were consistent with known properties of SCP release from B38. Next we selectively superfused the neuromuscular synapses with exogenous SCP to determine if this would occlude the effects of SCP released from B38 during a tetanus. We found that exogenous SCP dramatically reduced PTP for B38 but had little effect on PTP for B3. Thus our results support the hypothesis that physiological stimulation of B38 elicits PTP that is predominantly dependent on the release of SCP from its own terminals. They also demonstrate that the mechanisms underlying PTP can be very different for two motor neurons innervating the same target muscle. PMID- 11731542 TI - Integration of perspective and disparity cues in surface-orientation-selective neurons of area CIP. AB - We investigated the effects of linear perspective and binocular disparity, as monocular and binocular depth cues, respectively, on the response of surface orientation-selective (SOS) neurons in the caudal part of the lateral bank of the intraparietal sulcus (area CIP). During the single-unit recording, monkeys were required to perform the delayed-matching-to-sample (successive same/different discrimination) of discriminating surface orientation in stereoscopic computer graphics. Of 211 visually responsive neurons, 66 were intensively tested using the solid-figure stereogram (SFS) of a square plate with both disparity and perspective cues (D+P condition), and 62 of these were identified as SOS neurons for responding selectively to the orientation of stimuli. All these neurons were further tested using a solid figure with perspective cues alone (P-only condition), and 58% (36/62) of these showed selective response to the orientation of the stimuli. Of the 62 SOS neurons, 35 neurons were also tested using SFS with disparity cues alone (D-only condition) in addition to the D+P and P-only conditions. We classified these 35 neurons into four groups by comparing the response selectivity under the P-only and D-only conditions. More than one-half of these (19/35) were sensitive to both perspective and disparity cues (DP neurons), and nearly one-third (11/35) of these were sensitive to disparity cues alone (D neurons), but a few (2/35) were sensitive to perspective cues alone (P neurons). The remaining (3/35) neurons exhibited orientation selectivity only when both cues were present. In DP neurons, the preferred orientation under the D+P condition was correlated to those under the D-only and P-only conditions, and the response magnitude under the D+P condition was greater than those under the D only and P-only conditions, suggesting the integration of both cues for the perception of surface orientation. However, in these neurons, the orientation tuning sharpness under the D+P and D-only conditions was higher than that under the P-only condition, suggesting the dominance of disparity cues. After the single-unit recording experiments, muscimol was microinjected into the recording site to temporarily inactivate its function. In all three effective cases out of six microinjection experiments, discrimination of a three-dimensional (3D) surface orientation was impaired when disparity cues alone were present. In only one effective case, when a relatively large amount of muscimol was microinjected, discrimination of a 3D surface orientation was impaired even when both disparity and perspective cues were present. These results suggest that linear perspective is an important cue for representations of a 3D surface of SOS neurons in area CIP, although it is less effective than binocular disparity, and that both of these depth cues may be integrated in area CIP for the perception of surface orientation in depth. PMID- 11731543 TI - Alteration of medullary dorsal horn neuronal activity following inferior alveolar nerve transection in rats. AB - The effects of inferior alveolar nerve (IAN) transection on escape behavior and MDH neuronal activity to noxious and nonnoxious stimulation of the face were precisely analyzed. Relative thresholds for escape from mechanical stimulation applied to the whisker pad area ipsilateral to the transection were significantly lower than that for the contralateral and sham-operated whisker pad until 28 days after the transection, then returned to the preoperative level at 40 days after transection. A total of 540 neurons were recorded from the medullary dorsal horn (MDH) of the nontreated naive rats [low-threshold mechanoreceptive (LTM), 27; wide dynamic range (WDR), 31; nociceptive specific (NS), 11] and sham-operated rats with skin incision (LTM, 34; WDR, 30; NS, 23) and from the ipsilateral (LTM, 82; WDR, 82; NS, 31) and contralateral MDH relative to the IAN transection (LTM, 77; WDR, 82; NS, 33). The electrophysiological properties of these neurons were precisely analyzed. Background activity of WDR neurons on the ipsilateral side relative to the transection was significantly increased at 2-14 days after the operation as compared with that of naive rats. Innocuous and noxious mechanical evoked responses of LTM and WDR neurons were significantly enhanced at 2-14 days after IAN transection. The mean area of the receptive fields of WDR neurons was significantly larger on the ipsilateral MDH at 2-7 days after transection than that of naive rats. We could not observe any modulation of thermal responses of WDR and NS neurons following IAN transection. Also, no MDH neurons were significantly affected in the rats with sham operations. The present findings suggest that the increment of neuronal activity of WDR neurons in the MDH following IAN transection may play an important role in the development of the mechano-allodynia induced in the area adjacent to the area innervated by the injured nerve. PMID- 11731544 TI - Synaptic regulation of the slow Ca2+-activated K+ current in hippocampal CA1 pyramidal neurons: implication in epileptogenesis. AB - The slow Ca2+-activated K+ current (sI(AHP)) plays a critical role in regulating neuronal excitability, but its modulation during abnormal bursting activity, as in epilepsy, is unknown. Because synaptic transmission is enhanced during epilepsy, we investigated the synaptically mediated regulation of the sI(AHP) and its control of neuronal excitability during epileptiform activity induced by 4 aminopyridine (4AP) or 4AP+Mg2+-free treatment in rat hippocampal slices. We used electrophysiological and photometric Ca2+ techniques to analyze the sI(AHP) modifications that parallel epileptiform activity. Epileptiform activity was characterized by slow, repetitive, spontaneous depolarizations and action potential bursts and was associated with increased frequency and amplitude of spontaneous excitatory postsynaptic currents and a reduced sI(AHP.) The metabotropic glutamate receptor (mGluR) antagonist (S)-alpha-methyl-4 carboxyphenylglycine did not modify synaptic activity enhancement but did prevent sI(AHP) inhibition and epileptiform discharges. The mGluR-dependent regulation of the sI(AHP) was not caused by modulated intracellular Ca2+ signaling. Histamine, isoproterenol, and (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid reduced the sI(AHP) but did not increase synaptic activity and failed to evoke epileptiform activity. We conclude that 4AP or 4AP+Mg-free-induced enhancement of synaptic activity reduced the sI(AHP) via activation of postsynaptic group I/II mGluRs. The increased excitability caused by the lack of negative feedback provided by the sI(AHP) contributes to epileptiform activity, which requires the cooperative action of increased synaptic activity. PMID- 11731546 TI - Hand synergies during reach-to-grasp. AB - An emerging viewpoint is that the CNS uses synergies to simplify the control of the hand. Previous work has shown that static hand postures for mimed grasps can be described by a few principal components in which the higher order components explained only a small fraction of the variance yet provided meaningful information. Extending that earlier work, this study addressed whether the entire act of grasp can be described by a small number of postural synergies and whether these synergies are similar for different grasps. Five right-handed adults performed five types of reach-to-grasps including power grasp, power grasp with a lift, precision grasp, and mimed power grasp and mimed precision grasp of 16 different objects. The object shapes were cones, cylinders, and spindles, systematically varied in size to produce a large range of finger joint angle combinations. Three-dimensional reconstructions of 21 positions on the hand and wrist throughout the reach-to-grasp were obtained using a four-camera video system. Singular value decomposition on the temporal sequence of the marker positions was used to identify the common patterns ("eigenpostures") across the 16 objects for each task and their weightings as a function of time. The first eigenposture explained an average of 97.3 +/- 0.89% (mean +/- SD) of the variance of the hand shape, and the second another 1.9 +/- 0.85%. The first eigenposture was characterized by an open hand configuration that opens and closes during reach. The second eigenposture contributed to the control of the thumb and long fingers, particularly in the opening of the hand during the reach and the closing in preparation for object grasp. The eigenpostures and their temporal evolutions were similar across subjects and grasps. The higher order eigenpostures, although explaining only small amounts of the variance, contributed to the movements of the fingers and thumb. These findings suggest that much of reach-to-grasp is effected using a base posture with refinements in finger and thumb positions added in time to yield unique hand shapes. PMID- 11731545 TI - Cell-type-specific GABA responses and chloride homeostasis in the cortex and amygdala. AB - The GABA responses of fast-spiking (FS) interneurons and regular-spiking (RS) principal cells were studied using whole cell and perforated-patch recordings in slices of the basolateral amygdala, neo-, and perirhinal cortex. In these three areas, responses to exogenous and synaptically released GABA were abolished by GABA(A) receptor antagonists in FS cells but also included a GABA(B) component in RS cells. Moreover, E(GABA(A)) of FS and RS cells differed from the calculated E(Cl) (-61 mV), but in opposite direction (FS, -54 mV; RS, -72 mV). This was not due to a differential dialysis of FS and RS cells by the pipette solution because the discrepancy persisted when recordings were obtained with the perforated-patch clamp technique, using the cation-selective ionophore gramicidin. Moreover, pharmacological inhibition of cation-chloride cotransporters revealed that the differing E(GABA(A)) of FS and RS neurons arises from cell-type-specific chloride homeostatic mechanisms. Indeed, the prevalent regulators of the intracellular chloride concentration are cotransporters that accumulate chloride in FS cells and extrude chloride in RS neurons. Thus, our results suggest that in the basolateral amygdala as well as in the parietal and perirhinal cortices, FS interneurons are more excitable than principal cells not only by virtue of their dissimilar electroresponsive properties but also because they express a different complement of GABA receptors and chloride homeostatic mechanisms. PMID- 11731547 TI - Dopamine inhibition of evoked IPSCs in rat prefrontal cortex. AB - Rat prefrontal cortex (PFC) receives substantial dopamine (DA) input. This DA innervation appears critical for modulation of PFC cognitive functions. Clinical and experimental studies have also implicated DA in the pathogenesis of a number of neurological and psychiatric disorders including epilepsy and schizophrenia. However, the actions of DA at the cellular level are incompletely understood. Both inhibitory interneurons and pyramidal cells are targets of DA and may express different DA receptor types. Our recent findings suggest that DA can directly excite cortical interneurons and increase the frequency of spontaneous inhibitory postsynaptic currents (IPSCs). The present study was undertaken to determine the effect of specific DA receptor agonists on evoked (e) IPSCs. Visually identified pyramidal neurons were studied using whole cell voltage-clamp techniques. Bath application of DA 30 microM reduced IPSC amplitude to 80 +/- 4% (mean +/- SE) of control without any significant change in IPSC kinetics or passive membrane properties. The D1-like DA receptor agonist SKF 38393 reduced IPSC amplitude to 71.5 +/- 8%, whereas the D2-like specific agonist quinpirole has no effect on amplitude (94.5 +/- 5%). The D1-like receptor antagonist SCH 23390 prevented DA inhibition of IPSC amplitude (98.2 +/- 4%), whereas IPSCs were still reduced in amplitude (79.7 +/- 4%) by DA in the presence of the D2-like receptor antagonist sulpiride. DA increased significantly paired-pulse inhibition, whereas responses to puff applied GABA were unaffected. Addition of the PKA inhibitor H-8 blocked the effect of DA on IPSCs. These results suggest that DA can decrease IPSCs in layer II-III PFC neocortical pyramidal cells by activating presynaptic D1-like receptors. PMID- 11731548 TI - Direction tuning of inhibitory inputs to the turtle accessory optic system. AB - Neurons in turtle accessory optic system (basal optic nucleus, BON) were studied to compare excitatory and inhibitory visual inputs. Using a reduced in vitro brain stem preparation with the eyes attached, previous studies only showed a monosynaptic retinal input to the BON from direction-sensitive retinal ganglion cells that share a common preferred direction. Now using an intact brain stem preparation, not only did BON neurons display inhibitory postsynaptic potentials [IPSP(C)s] spontaneously, but IPSP(C)s were also evoked by visual pattern motion, they had their polarity reversed near the chloride equilibrium potential and they were blocked by the GABA(A) antagonist bicuculline. Because excitatory postsynaptic currents had reversal potentials >0 mV, BON cells were recorded using patch electrodes filled with QX-314 or Cs+ to measure the cell's direction tuning also at that higher reversal potential. For most of the BON neurons studied, their visual excitation and inhibition had a very similar preferred direction, indicating that both synaptic inputs were maximally active onto the same cell under the same stimulus conditions. These competing inputs may result from connections between the pretectum and accessory optic nuclei. Such synaptic interactions may serve a functional role in the visual processing necessary to create retinal slip signals for oculomotor control. PMID- 11731549 TI - Acute sensitization by NGF of the response of small-diameter sensory neurons to capsaicin. AB - We investigated acute sensitization by nerve growth factor (NGF) of the response of small-diameter (<30 microm) dissociated dorsal root ganglion (DRG) cells to brief repeated puffs of capsaicin as a model for thermal hyperalgesia induced by NGF. We have previously shown that placing NGF in the bath after an initial puff of capsaicin can completely overcome the tachyphylaxis normally observed in response to a second puff 10 min later, and this response is often substantially larger than the first. If tachyphylaxis is abolished by carrying out the experiment in Ca2+-free solution, NGF still elicits potentiation of the second puff. However, the amount of potentiation is considerably less than that observed when tachyphylaxis also takes place. Thus it is concluded that NGF has two effects: overcoming tachyphylaxis and potentiation. With three puffs of capsaicin separated by 10 min, we have found that the potentiation established after 10 min exposure to NGF is no longer evident 10 min after removal of NGF. In Ca2+-free solution the potentiation can last up to 1 h after removal of NGF. These results suggest that the initial behavioral sensitization elicited by NGF could result from a direct effect on the sensory neuron but that its later components most likely involve other mechanisms. We have also investigated the contribution of various second-messenger pathways in these actions of NGF by treating the cells with blockers of MAP kinase (PD98059), protein kinase A (PKA; PKAI14-22, H89), and PKC (Bisindolylmaleimide I). Surprisingly, PD98059, which previously has been shown to diminish the enhancement of capsaicin responses of dissociated neurons when exposed to NGF for several days, had no effect on the acute response to NGF; nor did the PKC inhibitor. However, PKA inhibitors reduced the capsaicin response of the cells to NGF (as determined from the NGF effect on tachyphylaxis). Consistent with these findings we confirmed that forskolin, a PKA activator, enhances the effect of NGF on the capsaicin response. The percentage of small cells sensitized by NGF under these conditions, as determined by its ability to reduce tachyphylaxis, was 64%. This suggests that about two-thirds of DRG cells <30 microm and sensitive to capsaicin express a functional trkA receptor. PMID- 11731550 TI - Neuronal bursting induced by NK3 receptor activation in the neonatal rat spinal cord in vitro. AB - Intracellular recording from lumbar motoneurons and extracellular recording from ventral roots of the neonatal rat isolated spinal cord were used to study the mechanisms responsible for the excitation mediated by NK3 tachykinin receptors. The selective NK3 agonists senktide or [MePhe7]neurokinin B induced a slow depolarization with superimposed oscillations (mean period +/- SD was 2.8 +/- 0.8 s) that, in the majority of cases, showed left-right alternation at segmental level and were synchronous between L2 and L5 of the same side. During agonist wash out (5-20 min) a delayed form of hyperexcitability emerged consisting of bursts lasting 8 +/- 2 s (average interburst interval 55 +/- 21 s) with superimposed oscillations usually with homosegmental alternation and heterosegmental synchronicity. Such bursting was accompanied by depression of GABAergic dorsal root potentials evoked by dorsal root stimulation and of the recurrent inhibitory postsynaptic potential recorded from motoneurons. Despite bursting, motoneuron membrane potential returned to baseline while input resistance was increased. Bursts were a network-dependent phenomenon triggered by previous NK3 receptor activation because bursting was suppressed by glutamate receptor antagonists and was insensitive to motoneuron membrane potential or subsequent application of an NK3 receptor antagonist. NK3 receptors operated synergistically with N-methyl-D-aspartate (NMDA) and 5-hydroxytryptamine (5-HT) to trigger fully alternating locomotor-like rhythms while NK3 receptor antagonism disrupted the same rhythm. In summary, in the neonatal rat spinal cord NK3 receptors could trigger rhythmic activity predominantly with alternation at segmental level but with synchronous coupling between ipsilateral motor pools. NK3 receptor activation could also facilitate fictive locomotor patterns induced by NMDA and 5-HT. PMID- 11731551 TI - Limits to the development of fast neuromuscular transmission in zebrafish. AB - Zebrafish embryos have small and slow miniature end-plate currents (mEPCs), whereas only a few days later larval mEPCs are an order of magnitude larger and faster, being among the fastest of all neuromuscular synapses. To identify the bases for these changes we compared, in embryos and larvae, the properties and distributions of acetylcholine (ACh) receptors (AChRs) and acetylcholinesterase (AChE) as well as the ultrastructure of the developing neuromuscular junctions (NMJs). To mimic synaptic release, patches of muscle membrane were exposed briefly (for 1 ms) to a saturating concentration (10 mM) of ACh. The AChR deactivation kinetics were twice as slow in embryos compared with larvae. In both embryos and larvae, AChRs demonstrated open channel block by millimolar ACh, and this was detected during mEPCs, indicating that a high concentration of ACh is released at immature and mature NMJs. AChR and AChE distributions were compared using the selective fluorescently conjugated labels alpha-bungarotoxin and fasciculin 2, respectively. In larvae, punctate AChR clusters were detected whereas junctional AChE staining was less intense than that found at adult NMJs. Transmission electron microscopy revealed immature nerve endings in embryos that were closely juxtaposed to the surrounding muscle cells, whereas mature larval NMJs had a wider synaptic cleft with a conspicuous basal lamina over a limited region of synaptic contact. Our results indicate that ACh is released at high concentrations at immature NMJs, but its clearance is prolonged and the AChRs are dispersed, resulting in a slow mEPC time course until a mature cleft appears with densely packed faster AChRs and abundant AChE. PMID- 11731552 TI - Amine modulation of the transient potassium current in identified cells of the lobster stomatogastric ganglion. AB - The pyloric network of the stomatogastric ganglion of the lobster Panulirus interruptus is a model system used to understand how motor networks change their output to produce a variety of behaviors. The transient potassium current (I(A)) shapes the activity of individual pyloric neurons by affecting their rate of postinhibitory rebound and spike frequency. We used two electrode voltage clamp to study the modulatory effects of dopamine (DA), octopamine (OCT), and serotonin (5-HT) on I(A) in the anterior burster (AB), inferior cardiac (IC), and ventricular dilator (VD) neurons of the pyloric circuit. DA significantly reduced I(A) in the AB and IC neurons and shifted their voltages of activation (V(act)) and inactivation (V(inact)) in a depolarized direction. These ionic changes contribute to the depolarization and increased firing rate of the AB and IC neurons produced by DA. Likewise, 5-HT significantly reduced I(A) and shifted V(inact) in the depolarized direction in the IC neuron, consistent with 5-HT's enhancement of IC firing. None of the amines evoked significant changes in I(A) in the VD neuron, suggesting that other currents mediate the amine effects on this neuron. PMID- 11731553 TI - Muscarine reduces calcium-dependent electrical activity in substantia nigra dopaminergic neurons. AB - The effect of muscarine on Ca2+ dependent electrical activity was studied in dopamine (DA) neurons located in the substantia nigra pars compacta (SNc) in brain slices from young rats, using sharp electrodes. In most DA neurons tested, muscarine (50 microM) reduced the amplitude of spontaneous oscillatory potentials and evoked Ca2+-dependent potentials recorded in the presence of TTX. Muscarine also reduced the amplitude of the slow afterhyperpolarization (sAHP) following action potentials in most DA neurons. These data suggest that muscarine reduces Ca2+ entry in SNc DA neurons. The reduction of the amplitude of the sAHP by muscarine in DA neurons may facilitate bursting initiated by glutamatergic input by increasing the frequency at which DA neurons can fire. The reduction of the sAHP via activation of muscarinic receptors in vivo may provide a mechanism whereby cholinergic inputs to DA neurons from the tegmental peduncular pontine nucleus could modulate dopamine release at dopaminergic targets in the brain. PMID- 11731554 TI - Properties of excitatory synaptic connections mediated by the corpus callosum in the developing rat neocortex. AB - Despite the major role of excitatory cortico-cortical connections in mediating neocortical activities, little is known about these synapses at the cellular level. Here we have characterized the synaptic properties of long-range excitatory-to-excitatory contacts between visually identified layer V pyramidal neurons of agranular frontal cortex in callosally connected neocortical slices from postnatal day 13 to 21 (P13-21) rats. Midline stimulation of the corpus callosum with a minimal stimulation paradigm evoked inward excitatory postsynaptic currents (EPSCs) with an averaged peak amplitude of 56.5 +/- 5 pA under conditions of whole cell voltage clamp at -70 mV. EPSCs had fixed latencies from stimulus onset and could follow stimulus trains (1-20 Hz) without changes in kinetic properties. Bath application of 2,3-dihydro-6-nitro-7-sulfamoyl benzo(F)quinoxaline (NBQX) abolished these responses completely, indicating that they were mediated by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors (AMPARs). Evoked responses were isolated in picrotoxin to yield purely excitatory PSCs, and a low concentration of NBQX (0.1 microM) was used to partially block AMPARs and prevent epileptiform activity in the tissue. Depolarization of the recorded pyramidal neurons revealed a late, slowly decaying component that reversed at approximately 0 mV and was blocked by D-2-amino-5 phosphonovaleric acid. Thus AMPA and N-methyl-D-aspartate receptors (NMDARs) coexist at callosal synapses and are likely to be activated monosynaptically. The peak amplitudes and decay time constants for EPSCs evoked using minimal stimulation (+/-40 mV) were similar to spontaneously occurring sEPSCs. Typical conductances associated with AMPA and NMDAR-mediated components, deduced from their respective current-voltage (I-V) relationships, were 525 +/- 168 and 966 +/ 281 pS, respectively. AMPAR-mediated responses showed age-dependent changes in the rectification properties of their I-V relationships. While I-Vs from animals >P15 were linear, those in the younger ( or =51-year-olds). The diagnostic codes used were based on the International Classification of Diseases (ICD)-6, 1956-1957; ICD-7, 1958-1967; ICD-8, 1968 1978; and ICD-9, 1979-1997. Over the period studied, this hospital was the primary inpatient provider for children in this area, and the only provider for uninsured children. Between 1960 and 1990, the racial makeup of the area remained stable, as did the percentage of blacks living at the poverty level. The raw number of asthma discharges, rate per 10 000 discharges of the same race, and rate per 100 000 population in Charleston County were tabulated for each age group and racial category. RESULTS: Over the time period examined, there has been a progressive increase in asthma hospitalizations in black individuals of all age groups and in whites under 18 years. The most striking increase has been in black children 0 to 18 years old (figure). The increase either as raw values or as a rate per 100 000 began around 1970, and was linear. This increase in black children discharged with asthma as a rate per 100 000 population was 20-fold: the rate increased from 18 in 1970 to 370 in 1997. Asthma discharges as a rate per 10 000 black children discharged increased by 24-fold from 1960 to 1997. Total discharges from the hospital increased from 49 000 to 128 000 per year over this period. Blacks made up only 28% of discharges in 1957, but that proportion increased to 56% in 1960 and remained relatively stable over the following 35 years. The increase seen in white children 0 to 18 years of age as a rate per 100 000 population was 5-fold and began around 1980. Both increases seem to be consistent over the time period studied, and continued to 1997. [figure: see text]. CONCLUSIONS: Among a predominantly poor black population living in a southern US city, there has been a steady increase in childhood asthma hospitalizations over the past 30 years. A significant although less dramatic rise has occurred in white children. Over this time period, although there have been many changes in lifestyle that could have contributed to this rise, there have been no major changes in housing conditions for poor patients. In addition, Medicaid coverage for children in South Carolina did not change significantly until 1999. The time course of these increases parallels increases reported in other Western populations, suggesting that there must be 1 or more common factors contributing to the rise. Many explanations have been offered for the changes in incidence and severity of asthma. The scale of the change, time course, and linearity of the increase in this study represent a challenge to many of the hypotheses proposed to explain this epidemic. PMID- 11731625 TI - Attitudes, practices, and preferences of pediatricians regarding initiation of hepatitis B immunization at birth. AB - OBJECTIVES: To explore practices and attitudes of pediatricians toward administration of the first dose of hepatitis B vaccine to infants, and to identify factors influencing the decision of pediatricians to initiate immunization at birth versus at 1 to 2 months of age. METHODS: A random sample of 600 pediatricians obtained from the American Academy of Pediatrics membership database was surveyed by mail. RESULTS: Three hundred eighty (68%) of the 563 pediatricians who were located responded to the survey. Of these 380 pediatricians, 279 provided routine immunizations to children. Of the 270 pediatricians who vaccinated children with hepatitis B vaccine and indicated their practice regarding the birth dose, 50% offered the first dose of hepatitis B vaccine at birth to all infants; the rest either offered the vaccine at birth only to infants of hepatitis B surface antigen-positive mothers and mothers whose serostatus is unknown, or did not offer the birth dose to any infants at all. Practicing in the inner city, working for a medical school or government hospital, and living in a state with universal immunization supply policies were associated with the respondent giving the birth dose. The strongest perceived barriers to giving the birth dose in the hospital were the difficulty tracking these vaccines (39%), the increased cost (27%), and the lack of reimbursement from insurance companies (26%). If a combination vaccine that includes hepatitis B; diphtheria, tetanus, pertussis (diphtheria and tetanus toxoids and acellular pertussis vaccine); and polio (inactivated poliovirus vaccine) antigens become available in the near future, then 38% of physicians who currently give the birth dose to all infants would prefer to wait until 2 months of age to initiate hepatitis B immunization. CONCLUSIONS: Efforts to achieve high implementation of hepatitis B birth dose administration may falter once a hepatitis B-containing pentavalent combination vaccine becomes available. Programmatic efforts should ensure prevention of perinatal hepatitis B virus transmission through universal prenatal hepatitis B surface antigen screening and immunoprophylaxis of high-risk newborn infants. PMID- 11731626 TI - Predictors of influenza virus vaccination status in hospitalized children. AB - OBJECTIVE: To determine predictors of influenza virus vaccination status in children who are hospitalized during the influenza season. METHODS: A cross sectional study was conducted among children who were hospitalized with fever between 6 months and 3 years of age or with respiratory symptoms between 6 months and 18 years of age. The 1999 to 2000 influenza vaccination status of hospitalized children and potential factors that influence decisions to vaccinate were obtained from a questionnaire administered to parents/guardians. RESULTS: Influenza vaccination rates for hospitalized children with and without high-risk medical conditions were 31% and 14%, respectively. For both groups of children, the vaccination status was strongly influenced by recommendations from physicians. More than 70% of children were vaccinated if a physician had recommended the influenza vaccine, whereas only 3% were vaccinated if a physician had not. Lack of awareness that children can receive the influenza vaccine was a commonly cited reason for nonvaccination. CONCLUSIONS: A minority of hospitalized children with high-risk conditions had received the influenza vaccine. However, parents' recalling that a clinician had recommended the vaccine had a positive impact on the vaccination status of children. PMID- 11731627 TI - Evaluation of administration of activated charcoal in the home. AB - OBJECTIVE: Activated charcoal (AC) is recognized as the treatment of choice for gastrointestinal decontamination after many ingestions. AC use in the home has been limited by concerns that parents would not administer it properly and that children would refuse to take AC. Previous descriptions of home administration have reported mixed results. METHODS: This was an 18-month consecutive case series of all patients for whom AC administration was recommended in the home. Data collected included AC availability in the home and/or a local pharmacy, success in administration, amount administered, time after ingestion to AC administration, difficulties in administration, adverse effects, age and gender of patient, substance involved in poisoning, and medical outcome. All cases were followed for at least 3 days after the ingestion. Patients who initially had home AC recommendation but who ultimately were treated in the emergency department (ED) served as a comparison group. RESULTS: Home administration of AC was recommended in 138 cases. A total of 115 individuals (83%) were treated with AC in the home, with no failures to administer AC. Reasons for failure to manage at home were 1) mother preferred ED (8 cases), 2) could not locate AC (7 cases), 3) pharmacy closed for the night (6 cases) and 4) no home telephone for follow-up (2 cases). Time to AC administration after ingestion was a mean of 38 minutes (+/ 18.3) for home treatment and 73 minutes (+/-18.1) for ED treatment. Ninety-five percent of home cases received AC in < or =60 minutes versus 33% for ED management. AC was in the home in 11 cases at the time of recommendation. The amount of AC administered was a mean of 12.1 g (standard deviation: 6.9) and a median of 12 g. Eight children (6.9%) who were treated at home vomited after AC versus 3 (13%) who received ED treatment. No aspirations or complications occurred. CONCLUSIONS: AC can be administered successfully by the lay public in the home. Home use of AC significantly reduces the time to AC administration. PMID- 11731628 TI - Newborns with suspected occult spinal dysraphism: a cost-effectiveness analysis of diagnostic strategies. AB - OBJECTIVE: To assess the clinical and economic consequences of different diagnostic strategies in newborns with suspected occult spinal dysraphism. METHODS: A decision-analytic model was constructed to project the cost and health outcomes of magnetic resonance imaging (MRI), ultrasound (US), plain radiographs, and no imaging in newborns with suspected occult spinal dysraphism. Morbidity and mortality rates of early versus late diagnosis of dysraphism and the sensitivity and specificity of MRI, US, and plain radiographs were obtained from the literature. Cost estimates were obtained from a hospital cost accounting database and from the Medicaid fee schedule. RESULTS: We found that the choice of imaging strategy depends on the underlying risk of occult spinal dysraphism. In low-risk children with intergluteal dimple or newborns of diabetic mothers (pretest probability: 0.3%-0.34%), US was the most effective strategy with an incremental cost-effectiveness ratio of $55 100 per quality-adjusted life year gained. For children with lumbosacral dimples, who have a higher pretest probability of 3.8%, US was less costly and more effective than the other 3 strategies considered. In intermediate-risk newborns with low anorectal malformation (pretest probability: 27%), US was more effective and less costly than radiographs and no imaging. However, MRI was more effective than US at an incremental cost-effectiveness of $1000 per quality-adjusted life year gained. In the high-risk group that included high anorectal malformation, cloacal malformation, and exstrophy (pretest probability: 44%-46%), MRI was actually cost-saving when compared with the other diagnostic strategies. For the intermediate-risk group, we found our analysis to be sensitive to the costs and diagnostic performances (sensitivity and specificity) of MRI and US. Lower MRI cost or greater MRI diagnostic performance improved the cost-effectiveness of the MRI strategy, whereas lower US cost or greater US diagnostic performance worsened the cost-effectiveness of the MRI strategy. Therefore, individual or institutional expertise with a specific diagnostic modality (MRI versus US) may influence the optimal diagnostic strategy. CONCLUSIONS: In newborns with suspected occult dysraphism, appropriate selection of patients and diagnostic strategy may increase quality-adjusted life expectancy and decrease cost of medical work-up. PMID- 11731629 TI - Economic impact of an infection control education program in a specialized preschool setting. AB - OBJECTIVE: To assess the economic impact, from a societal perspective, of a multidimensional infection control education program (ICEP) in a preschool for children with Down syndrome. METHODS: Krilov et al implemented a comprehensive ICEP in a specialized preschool setting and reported a significant decrease in medical resource utilization and days absent from school. Clinical and economic data from Krilov et al and other sources were incorporated into a health-state transition (Markov) decision analysis model that estimated annual expected costs for the baseline and intervention years. Procedure and diagnosis codes were assigned to all physician office visits, emergency department visits, hospitalizations, and laboratory and diagnostic tests. Cost estimates then were derived using 1999 national reimbursement schedules and other sources. Productivity losses for parents were estimated using national wage rates. The costs of the ICEP were compared with the reduction in the costs of illness (direct medical costs plus costs associated with lost parental working time). The outcomes measured were mean annual costs of illness per child, total annual ICEP costs, and net annual costs or savings. RESULTS: With a comprehensive ICEP, the mean costs of illness in the baseline year was $1235 per child, of which 68% and 14% were for productivity losses and physician visits, respectively. In the intervention year, the mean costs of illness per child was $615, of which 71% and 20% were for productivity losses and physician visits, respectively. The cost of the preexisting infection control (IC) practices in place at the onset of the study (baseline year) was $716. The comprehensive ICEP cost (intervention year) was $75 627, 92% of which was spent to hire a cleaning service to decontaminate toys 3 times per week. When a secondary analysis was performed to reflect a less intensive ICEP in a nonspecialized preschool setting, the mean costs of illness in the baseline and intervention years were $962 and $614 per child, respectively, representing a total annual cost-of-illness savings of $13 224 for the 38 children who participated in the study by Krilov et al. The annual incremental cost of the less intensive ICEP was $2371; therefore, the estimated net annual savings of the less intensive ICEP in a nonspecialized preschool was $10 853. CONCLUSIONS: This study suggests that the reduction in the costs of illness could more than offset the cost of implementing a multidimensional ICEP in a preschool setting. PMID- 11731631 TI - Cardiovascular anomalies in patients diagnosed with a chromosome 22q11 deletion beyond 6 months of age. AB - OBJECTIVE: Cardiovascular anomalies are present in 75% to 80% of patients with a chromosome 22q11 deletion. In the majority of cases, the cardiovascular defect becomes evident in the neonatal period and is often the initial manifestation of the chromosome 22q11 deletion syndrome. However, a 22q11 deletion may also be associated with cardiovascular defects that are less obvious, such as a vascular ring, which may not be diagnosed until the patient is older. The objective of this study was to determine the frequency and types of cardiovascular anomalies in patients diagnosed with a chromosome 22q11 deletion beyond 6 months of age. METHODS: We studied 29 patients diagnosed with a chromosome 22q11 deletion at a median age of 6.2 years (9 months to 45 years) who were subsequently referred for cardiovascular evaluation. Comprehensive cardiologic evaluation was performed, with transthoracic echocardiography (N = 28) and/or magnetic resonance imaging (N = 6), including imaging of the aortic arch. The frequency of cardiovascular anomalies diagnosed in these patients and the need for intervention were assessed. RESULTS: Cardiovascular anomalies were detected in 11 (38%) patients: 3 with a vascular ring formed by a right aortic arch with an aberrant left subclavian artery and left-sided ligamentum arteriosum, 3 with a right aortic arch with mirror-image branching of the brachiocephalic arteries (no vascular ring; 1 with a patent ductus arteriosus), 4 with a left aortic arch with an aberrant right subclavian artery (no vascular ring; 1 with a patent ductus), and 1 with a left superior vena cava draining to the coronary sinus. The median age at diagnosis in these 11 patients was 3 years (9 months to 28 years). The remaining 18 patients had normal cardiovascular anatomy. All 3 patients with vascular rings subsequently underwent surgical repair, and 1 patient with a ductus arteriosus underwent transcatheter coil occlusion. CONCLUSIONS: The frequency of cardiovascular anomalies necessitating intervention in patients referred for cardiovascular evaluation after diagnosis of a chromosome 22q11 deletion beyond 6 months of age is 14% in our experience. Routine screening for cardiovascular anomalies, including echocardiography and other imaging studies to identify the laterality and branching pattern of the aortic arch, is indicated in patients diagnosed with 22q11 deletion beyond 6 months of age and is particularly critical for patients with respiratory or feeding disorders. PMID- 11731630 TI - Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae infection in pediatric private practice. AB - BACKGROUND: Universal screening of sexually active adolescents for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) has been recommended in settings in which prevalence is 2% or greater. Although believed to be above 2%, the prevalence of CT and GC infection in private practice settings has not been clearly established and may affect screening practices. OBJECTIVES: To determine CT and GC infection prevalence in 2 pediatric private practices. DESIGN: Cross sectional study. SETTING: Two pediatric private practices in suburban North Carolina. PATIENTS: Convenience sample of patients aged 15 to 24 years who were seen from August 1998 through June 1999. MAIN OUTCOME MEASURES: Prevalence of CT and GC infection. RESULTS: Of 1114 eligible patients, 803 (72%) completed questionnaires and provided urine specimens tested for CT and GC infection using ligase chain reaction assays. Mean age was 17.1 years (standard deviation: 1.8). Most participants were female (63%), white (87%), and from highly educated families (64% of their mothers graduated from college). Sexual activity was reported by 41%. Prevalence of CT infection in all participants was 0.9% (confidence interval [CI]: 0.4%-1.8%); in sexually active participants, 2.1% (CI: 0.9%-4.3%); in sexually active females, 2.7% (CI: 1.0%-5.7%); and in sexually active males, 0.9% (CI: 0.0%-5.1%). One case of GC infection was found. CONCLUSIONS: The prevalence of CT and GC infection in this private practice population was much lower than reported in other settings. Screening recommendations may need to be reassessed if other low prevalence populations are found. PMID- 11731632 TI - Defining the impact of hemophilia: the Academic Achievement in Children with Hemophilia Study. AB - OBJECTIVES: We characterized a population-based cohort of school-aged children with severe hemophilia with respect to type of treatment, on-demand versus prophylaxis, and frequency of bleeding episodes in the year before enrollment. We also investigated the association between hemophilia-related morbidity, measured by number of bleeding episodes in the year before enrollment, and academic performance after adjustment for other factors known to have an effect on achievement. Finally, we explored the mechanisms for the association between bleeding episodes and academic achievement. STUDY DESIGN: This study was a multicenter investigation of boys 6 to 12 years old with severe factor VIII deficiency (clotting factor level <2%) receiving care in US hemophilia treatment centers. Children with a history of inhibitor, severe developmental disorder, significant psychiatric disorder, or insufficient fluency in English were excluded from the study. On-demand treatment was defined as administration of clotting factor on the occurrence of a bleeding episode. Prophylactic therapy was defined as a course of regular infusions for >2 months with a goal of preventing bleeding episodes. Academic achievement was measured by the Wechsler Individual Achievement Test. Quality of life was measured by the Child Health Questionnaire. Of particular interest was the Physical Summary (PhS) measure of the Child Health Questionnaire. The type of information captured by the PhS includes limitations in physical activity, limitations in the kind or amount of schoolwork or social activities the child engaged in, and presence of pain or discomfort. RESULTS: One hundred thirty-one children were enrolled, a median center recruitment rate of 77%. The mean age of the participants was 9.6 years, and approximately half of the participants had completed less than the fourth grade at the time of enrollment. Sixty-two percent of the children were on prophylaxis at enrollment, and 9% had previously been on prophylaxis but were currently on on-demand therapy. Two groups were defined: ever treated with prophylaxis and never treated with prophylaxis. For those ever treated, treatment duration ranged from 2.7 months to 7.7 years, with one half of the children treated with prophylaxis for >40% of their lifetimes; 29% had always been on on-demand therapy. Children in both treatment groups were similar with respect to age, clotting factor level, parents' education, and IQ. The median number of bleeding episodes experienced in the year before enrollment for the cohort as a whole was 12. The median number of bleeding episodes in children on prophylaxis at enrollment was significantly lower than in children on on-demand therapy (6 vs 25.5). The mean achievement scores were within the average range of academic performance: reading, 100.4; mathematics, 101.6; language, 108.1; writing, 95.4; and total achievement, 102.5. When children were categorized as above or below the study group median by number of bleeding episodes, those who had a low number of bleeding episodes (< or =11) had better total achievement (104.4 vs 100.6) and mathematics (103.6 vs 99.6) than children in the higher bleeding episode category (> or =12) after adjusting for child's IQ and parents' education. Treatment with prophylaxis per se was not associated with better test scores, but children who had been treated on a regimen of long-term prophylaxis (>40% of lifetime) and reported < or =11 bleeding episodes in the year before enrollment had significantly higher scores in total achievement (104.9 vs 100.6), mathematics (105.2 vs 99.6), and reading (104.0 vs 98.6) than all other children reporting > or =12 bleeding episodes in the same time period. Increased school absenteeism and hemophilia-related limitations in physical functioning among children with greater frequency of bleeding episodes were proposed as the mechanisms for lower scores. The number of bleeding episodes was positively correlated with school absenteeism (Spearman correlation = 0.23), and children with more school absences had lower scores in mathematics, reading, and total achievement, even after adjusting for the child's IQ and parents' education. Children with fewer bleeding episodes also had better PhS scores than children in the high bleeding episode category (48.4 vs 41.3). The mean PhS for children in the low bleeding episode group (48.4) was similar to that of the general US population (50), but the mean PhS for children in the higher bleeding episode group was almost a full standard deviation lower than the mean for the general US population. PhS scores were positively related to reading and total achievement scores after adjusting for IQ and parents' education. Of interest and concern was a group of children who were reportedly being treated with prophylaxis during the year before enrollment (N = 18) but whose bleeding events were not optimally suppressed. These children were 3 times as likely (33.3% vs 11.1%) to be receiving < or =2 infusions per week as children on prophylaxis who reported < or =11 bleeding episodes during the same period. A review of the sites of bleeding reported for the 18 children revealed that 12 (66.6%) experienced > or =25% of their bleeding episodes in the same joint. CONCLUSIONS: Each child should have the opportunity to achieve his or her potential. Control of a chronic disorder must include this important goal as well as the more commonly identified medical outcomes. This study has identified an important association between the number of bleeding episodes experienced and academic achievement in a cohort of school-aged children with severe hemophilia. The data support the assertion that therapeutic care programs in this population must not be evaluated only in terms of financial cost to achieve adequate musculoskeletal outcomes. Also significant are the individual and societal benefits of increased academic accomplishments if adequate suppression of hemorrhagic events can be attained. The number of bleeding episodes experienced, regardless of treatment regimen, should be followed to optimize the child's academic outcome. PMID- 11731633 TI - Swallowing function and medical diagnoses in infants suspected of Dysphagia. AB - OBJECTIVE: There has been an increase in infant swallowing disorders as a result of improved survival for infants born prematurely or with life-threatening medical disorders. These infants often have multiple health issues and an increased risk of respiratory complications. However, there is little understanding of the biomechanics of infant swallowing disorders. Therefore, the objectives of this study were to determine 1) the percentage of dysphagic infants who experience laryngeal penetration, aspiration, or nasopharyngeal backflow; 2) reasons for laryngeal penetration/aspiration; 3) whether infants with laryngeal penetration/aspiration clear their airway; and 4) the relationship between swallowing disorders and medical diagnoses. METHODS: Patients included 43 infants who were referred for videofluoroscopic swallowing studies in a university affiliated pediatric medical center. Medical charts were reviewed. The videofluoroscopic swallowing studies were recorded on videotape, and each swallow was analyzed for laryngeal penetration, aspiration, nasopharyngeal backflow, cough, airway clearance, and reason for penetration/aspiration. Statistics included chi2 for nonparametric data and measures of central tendency for numeric/timing data. RESULTS: More than half of the infants experienced laryngeal penetration, aspiration, or nasopharyngeal backflow; however, the first occurrence of these events was after multiple swallows. Only 3 infants experienced laryngeal penetration and aspiration on the first swallow and all 3 had an absent pharyngeal response. Premature infants experienced significantly more nasopharyngeal backflow. Material in the pyriform sinuses before pharyngeal swallowing was associated with penetration/aspiration. In episodes of laryngeal penetration, all patients were able to clear their airway during the swallow without a cough. Almost all infants (8 of 9) who aspirated did not cough or clear their airway. CONCLUSIONS: This study demonstrated that most infants suspected of dysphagia showed overt abnormalities: laryngeal penetration, aspiration, and/or nasopharyngeal backflow on the videofluoroscopic swallowing study. Most of these infants did not demonstrate abnormalities in the first few swallows but displayed deterioration in swallowing function as they continued to feed. Thus, radiographic assessments in infants must examine multiple swallows. The high incidence of silent aspiration demonstrates the necessity of a videofluoroscopic assessment to evaluate swallowing function in these infants. PMID- 11731634 TI - Analysis of the musculoskeletal system in children and adolescents receiving anticonvulsant monotherapy with valproic acid or carbamazepine. AB - OBJECTIVE: To examine bone development in children and adolescents who have uncomplicated idiopathic epilepsy and had received monotherapy with carbamazepine or valproic acid for at least 1 year. METHODS: Thirty-nine patients from 6 to 19 years of age (18 girls) were studied. Total bone mineral content (BMC) and trabecular volumetric bone mineral density were measured at the distal radius using peripheral quantitative computed tomography. Maximum isometric grip force was determined with a standard dynamometer. Alkaline phosphatase activity and deoxypyridinoline (a marker of bone resorption) were assessed in serum and urine, respectively. RESULTS: Trabecular volumetric bone mineral density was significantly decreased in the entire group (z score mean +/- standard deviation: -0.62 +/- 1.04) and in the subgroup using valproic acid (-0.75 +/- 1.18). In the carbamazepine subgroup, there was a similar but nonsignificant trend (-0.50 +/- 0.90). Total BMC and isometric maximum grip force were normal in the entire study population (0.10 +/- 1.22) and in the 2 subgroups. The relationship between BMC and grip force was similar between patients and healthy participants. Urinary levels of deoxypyridinoline were significantly elevated above normal in the whole study population (1.35 +/- 2.00) and in both the valproic acid and the carbamazepine subgroups. CONCLUSIONS: Bone turnover can be increased, but bone mass is adequate in children and adolescents who have uncomplicated idiopathic epilepsy and who receive monotherapy with carbamazepine or valproic acid. PMID- 11731635 TI - Comparison of body weight and height of Israeli schoolchildren with the Tanner and Centers for Disease Control and Prevention growth charts. AB - OBJECTIVE: To examine the suitability of the growth charts that are currently used in Israel and consider their replacement with a recent standard. In a sample of schoolchildren, the current Tanner and Whitehouse standards were compared with the new growth charts from the Centers for Disease Control and Prevention (CDC). METHOD: A total of 746 Israeli healthy children (368 boys and 378 girls) aged 6 to 14 years were sampled from 2 super-regional schools (from the Jewish population). Height was measured using the Leicester height measure, and weight was measured using scales calibrated by the researchers. RESULTS: In height, 75% of the boys and 81% of the girls fell in the 10th to 90th percentile range of the Tanner standards. In weight, 71% of the boys and 81% of the girls were in the 10th to 90th percentile range. Israeli boys were taller (chi2(1) = 30.53) and heavier (chi2(1) = 29.39), and girls were taller (chi2(1) = 13.81) than predicted by the Tanner standards. The CDC 10th to 90th percentile range included 81% of the boys and 83% of the girls for height and 78% of the boys and 81% of the girls for weight. The girls in our sample were slightly shorter (chi2(1) = 11.87) and lighter (chi2(1) = 9.52) than predicted from the CDC charts. Nevertheless, neither boys' nor girls' body mass index measures (kg/h2) differed from the CDC charts. CONCLUSIONS: Our data suggest that the CDC growth charts are adequate for assessing Israeli children and slightly better than the current standard. Thus, in the absence of local standards, we recommend the use of the revised CDC growth charts, which include the additional body mass index measure. PMID- 11731636 TI - Trends in booster seat use among young children in crashes. AB - INTRODUCTION: Booster seat use in the United States is extremely low among 4- to 8-year-old children, the group targeted for their use. However, more recent attention has been paid to the role of booster seats for children who have outgrown their forward-facing child safety seat. In particular, several states are currently considering upgrades to their child restraint laws to include the use of booster seats for children over 4 years of age. OBJECTIVE: To examine recent trends in booster seat use among children involved in automobile crashes in 3 large regions of the United States. DESIGN: This study was performed as part of the Partners for Child Passenger Safety project, an ongoing, child-specific crash surveillance system that links insurance claims data to telephone survey and crash investigation data. All crashes occurring between December 1, 1998, and November 30, 2000, involving a child occupant between 2 to 8 years of age riding in a model year 1990 or newer vehicle reported to State Farm Insurance Companies from 15 states and Washington, DC, were eligible for this study. A probability sample of eligible crashes was selected for a telephone survey with the driver of the vehicle using a previously validated instrument. The study sample was weighted according to each subject's probability of selection, with analyses conducted on the weighted sample. RESULTS: The weighted study sample consisted of 53 834 children between 2 to 8 years old, 11.5% of whom were using a booster seat at the time of the crash. Booster seat use peaked at age 3 and dropped dramatically after age 4. Over the period of study, booster seat use among 4- to 8-year-olds increased from 4% to 13%. Among 4-year-olds specifically, booster use increased from 14% to 34%. Among children using booster seats, approximately half used shield boosters and half used belt-positioning boosters. CONCLUSIONS: Although overall booster seat use among the targeted population of 4- to 8-year old children remains low, significant increases have been noted among specific age groups of children over the past 2 years. These data may be useful to pediatricians, legislators, and educators in efforts to target interventions designed to increase appropriate booster seat use in these children. PMID- 11731637 TI - Evaluation of resuscitation skills in new residents before and after pediatric advanced life support course. AB - OBJECTIVE: Acquisition of resuscitation skills taught in advanced resuscitation courses has not been evaluated. We sought to determine the interobserver reliability of a resuscitation performance skills checklist to assess resident performance of bag mask ventilation, intubation, defibrillation, and intraosseous vascular access, and to measure skill acquisition by entering residents after a pediatric advanced life support (PALS) course. DESIGN: The resuscitation skills of all 39 pediatric R1's in 2 university-based training programs were assessed immediately before and after completion of a PALS course just before starting residency. Independent observers scored and timed resident performance of bag mask ventilation, endotracheal intubation, intraosseous access, and defibrillation. Scores before and after the PALS course were compared. Four independent observers scored 4 residents' videotaped skills performance. Observers' scores for each resident were compared. RESULTS: Successful performance improved for bag mask ventilation from 62% to 97% after the PALS course; for successful endotracheal intubation, from 64% to 90%; for successful intraosseous needle placement, from 54% to 92%; and for successful defibrillation, from 77% to 97%. Interobserver reliability was high for continuous and noncontinuous variables. CONCLUSIONS: New residents demonstrated significant acquisition of pediatric resuscitation skills immediately after completion of the PALS course. The skills performance checklist has excellent interobserver reliability and is a useful tool for evaluation of other training venues. PMID- 11731638 TI - Delivery of pediatric after-hours care by call centers: a multicenter study of parental perceptions and compliance. AB - BACKGROUND: Despite the rapid growth of centralized call centers to provide after hours triage to patients of multiple providers, little is known about the perceptions of parents regarding this type of care and their compliance with triage disposition recommendations. DESIGN/METHODS: From August through September 1999, randomized samples of after-hours calls were selected each day from computerized records at 4 pediatric call centers at 1) Children's Hospital, Denver, Colorado; 2) Children's Hospital of Philadelphia, Philadelphia, Pennsylvania; 3) Rainbow Babies and Children's Hospital, Cleveland, Ohio; and 4) All Children's Hospital, St Petersburg, Florida. All participating call centers use the same triage software. Calls were randomly selected to yield at least 250 callers with nonurgent dispositions and 100 with urgent dispositions from each site. Telephone surveys to callers were conducted by an external survey unit 3 to 7 days after the call to the call center. RESULTS: Surveys were completed for 70.5% of those sampled (N = 1561). Parents indicated they were very satisfied or satisfied with aspects of care received from 92.6% (waiting time) to 99.4% (nurse courteousness) of the time. Satisfaction did not differ by site or by recommended disposition of the index call. Most parents (65.2%) reported no preference about speaking with a physician or nonphysician for after-hours care, whereas 27.7% preferred to speak with a physician. Usually speaking with a physician during office hours (odds ratio [OR]: 1.48), feeling it was important that provider knows child's medical history (OR: 3.47), and respondent having an educational level of college graduate or higher (OR: 1.30) were significant predictors of preferring to speak with a physician. Of the 37.0% (N = 723) of parents who reported any change in their relationship with their primary provider as a result of the after-hours call center, 95.7% (N = 691) assessed the change to be positive. Reported compliance with the call center disposition recommendation was 83.3% for urgent referral, 41.0% for next day, 4.5% for visit at a later time, and 78.2% for home care. The major reason given by parents for noncompliance was reporting that they heard a different disposition (76.9% for urgent to 100% for visit at a later time). CONCLUSIONS: Parental satisfaction with pediatric call centers was uniformly high in 4 different geographic locations, and almost all parents who reported any effect on their relationship with their primary provider assessed it as positive. Compliance with recommendations for urgent evaluation or home care was relatively high but for intermediary dispositions was low. In most cases in which noncompliance occurred, parents reported hearing a different disposition. Additional study is needed to clarify whether noncompliance, especially in cases in which an urgent recommendations was made, is attributable to poor nurse communication of the recommended disposition, parental misinterpretation, or parental difference of opinion. PMID- 11731639 TI - Childhood vaccinations, vaccination timing, and risk of type 1 diabetes mellitus. AB - OBJECTIVES: To evaluate suggested associations between childhood vaccinations, particularly against hepatitis B and Haemophilus influenzae type b, and risk of developing type 1 diabetes; and to determine whether timing of vaccination influences risk. METHODS: We conducted a case-control study within 4 health maintenance organizations (HMOs) that participate in the Vaccine Safety Datalink project of the Centers for Disease Control and Prevention. Study eligibility was restricted to children who met the following criteria: 1) born during 1988 through 1997; 2) HMO member since birth; 3) continuously enrolled for first 6 months of life; and 4) at least 12 months of HMO membership before diabetes incidence date (or index date for controls) unless incidence date was before 12 months of age. All 4 HMOs maintain registries of their members who have diabetes, and we used the registries to identify potential cases of diabetes. We conducted chart reviews to verify that potential cases met the World Health Organization epidemiologic case definition for type 1 diabetes mellitus (ie, a physician's diagnosis of diabetes plus treatment with daily insulin injections). We defined the incidence date of diabetes as the first date that the child received a diagnosis of diabetes. We attempted to match 3 controls to each case. Controls had the same eligibility criteria as cases and were matched to individual cases on HMO, sex, date of birth (within 7 days), and length of health plan enrollment (up to the incidence or index date). The index date for controls was defined as the incidence date of the case to which the control was matched. Chart abstraction was performed by trained chart abstractors using standardized forms. In addition to complete vaccination histories, the chart abstraction forms for both cases and controls included information on sociodemographic characteristics, selected medical conditions, history of breastfeeding, and family medical history. We used conditional logistic regression to estimate the odds ratio (OR) of diabetes associated with vaccination, with vaccine exposure defined as before the diabetes incidence date (or index date for controls). RESULTS: Two hundred fifty-two confirmed cases of diabetes and 768 matched controls met the study eligibility criteria. The OR (95% confidence interval) for the association with type 1 diabetes was 0.28 (0.07-1.06) for whole cell pertussis vaccine (predominantly in combination as diphtheria, tetanus toxoids and pertussis vaccine), 1.36 (0.70-2.63) for measles-mumps-rubella, 1.14 (0.51-2.57) for Haemophilus influenzae type b, 0.81 (0.52-1.27) for hepatitis B vaccine, 1.16 (0.72-1.89) for varicella vaccine, and 0.92 (0.53-1.57) for acellular pertussis containing vaccines. Compared with children who had not received hepatitis B vaccine, the OR of diabetes was 0.51 (0.23-1.15) for children vaccinated at birth and 0.86 (0.54-1.35) for those first vaccinated against hepatitis B at 2 months of age or later. Race and ethnicity and family history of diabetes were independently associated with risk of type 1 diabetes, but adjustment for these factors did not materially alter the ORs for any of the vaccines. CONCLUSIONS: In this large, population-based, case-control study, we did not find an increased risk of type 1 diabetes associated with any of the routinely recommended childhood vaccines. Our study adds to previous research by providing data on newer vaccines, including hepatitis B, acellular pertussis, and varicella vaccines. For the older vaccines, our results are generally in agreement with previous studies in not finding any increased risks. Ours is the first epidemiologic study to evaluate the possibility that timing of vaccination is related to risk of clinical diabetes in children. Our results on hepatitis B vaccine do not support the hypothesis; risk of type 1 diabetes was not different between infants vaccinated at birth and those who received their first vaccination later in life. The results of our study and the preponderance of epidemiologic evidence do not support an association between any of the recommended childhood vaccines and an increased risk of type 1 diabetes. Suggestions that diabetes risk in humans may be altered by changes in the timing of vaccinations also are unfounded. PMID- 11731640 TI - Eosinophilic cystitis in a 4-year-old boy: successful long-term treatment with cyclosporin A. AB - A 4-year-old Jewish boy presented with dysuria, urinary dribbling, increased urinary frequency, and new onset of diurnal enuresis. An infiltrating solid mass involving the entire bladder wall was found. Biopsy revealed "tumor-forming" eosinophilic cystitis, a rare bladder lesion of unclear cause. Antitoxocariasis treatment was unsuccessful. High-dose corticosteroids failed. The child's clinical condition and bladder sonographic findings continued to deteriorate. Treatment with cyclosporin A was given for 8 months, with a complete clinical, radiologic, and histopathologic cure and no side effects. Two years of follow-up showed a complete recovery. PMID- 11731641 TI - Neonatal aortic arch thrombosis as a result of congenital cytomegalovirus infection. AB - Thrombotic disease is rare in neonates. The main risk factors at this age are perinatal asphyxia, maternal diabetes, sepsis, polycythemia, dehydration, a low cardiac output, and in primis the catheterization of central lines. Another important risk factor is inherited thrombophilia. Arterial thrombosis is even more rare than venous thrombosis and less related to most of the risk factors listed above; it occurs more frequently in the iliac, femoral, and cerebral arteries but very rarely in the aorta. Most of the described cases of aortic thrombosis are associated with the catheterization of an umbilical artery and involve the descending tract and the renal arteries; very few relate to the ascending tract and the aortic arch. The possible role of virus-induced primary vascular endothelium damage in the etiopathogenesis of neonatal arterial thrombosis has been previously hypothesized. Herpesviruses, particularly human cytomegalovirus (HCMV), can infect endothelial cells and directly damage intact vascular endothelium, altering its thromboresistant surface as a result of procoagulant activity mediated by specific viral surface phospholipids, necessary for the coagulation enzyme complex assembly that leads to thrombin generation. We describe a case of congenital aortic arch thrombosis. The clinical, laboratory, and virologic pictures; the anatomopathologic findings (fully compatible with viral infection); the detection of HCMV in various tissues (including the aorta); and the absence of other causes of aortic thrombosis make it possible to attribute the case to a severe congenital HCMV infection with multiple organ involvement, after the primary infection of the mother. The hemostatic system disorders and hemodynamic disturbances related to viral cardiac damage explain the clinical features of the case and indicate that congenital HCMV infection should be included among the causes of neonatal aortic thrombosis. PMID- 11731642 TI - Immunologic and epidemiologic experience of vaccination with a monocomponent pertussis toxoid vaccine. AB - Pertussis re-emerged in Sweden with a cumulative incidence of about 60% during the first 10 years of life, when the locally produced cellular vaccine lost its efficacy around 1970 and general vaccination was discontinued in 1979. The epidemiology, clinical features, and immunology of pertussis and a monocomponent pertussis toxoid vaccine were studied in Goteborg, Sweden. After phase 1 and 2 studies, a randomized, double-blind, placebo-controlled trial of pertussis toxoid (PTox), compounded with diphtheria and tetanus toxoids, was administered to 3450 children according to the Swedish schedule at 3, 5, and 12 months of age. After a mean follow-up of 18 months, the efficacy was 71% overall and 75% in household contacts, respectively. A statistically significant correlation was found between the level of PTox-induced antibodies and protection against pertussis. As observed with cellular and with multicomponent acellular vaccines, PTox reduced the severity of disease and the percent of children with positive cultures. Furthermore, vaccination reduced the transmission of Bordetella pertussis to household contacts in the vaccinees compared with the controls who received only diphtheria and tetanus toxoids. Patients with culture-verified Bordetella parapertussis infection reacted with antibodies to pertactin and to filamentous hemagglutinin but not to pertussis toxin, and some subsequently developed pertussis. The antibody responses of patients with pertussis to the surface polysaccharides of B pertussis and to B parapertussis were cross-reactive serologically. Serosurveys showed that only antibodies to pertussis toxin were related to the occurrence of pertussis in the general population: antibodies to filamentous hemagglutinin and pertactin were probably stimulated by antigens of other bacteria as well as Bordetellae. Mass vaccination of Goteborg children born in the 1990s was started in 1995. In February 1999, about 55% had been vaccinated and both B pertussis and pertussis decreased significantly in individuals of all ages (herd immunity). Similar to diphtheria, PTox-induced immunity to pertussis occurs both on an individual and community basis. The apparent greater efficacy of multicomponent acellular pertussis vaccines compared with monocomponent PTox was proposed to be an artifact created when the diagnosis of pertussis was made by the serologic criteria of the World Health Organization only. Our conclusion is that PTox is both an essential and alone sufficient antigen in acellular pertussis vaccines. PMID- 11731643 TI - The Bloxsom air lock: a historical perspective. AB - In 1950, Allan P. Bloxsom (1901-1991), a pediatrician at the St Joseph Hospital in Houston, introduced his positive pressure oxygen air lock (AL) for the delivery room resuscitation of the asphyxiated newborn. The infant's entire body was placed into a cylindrical steel chamber that was tightly sealed and infused with warmed humidified 60% oxygen. The positive pressure within the AL was cycled between 1 and 3 lb/in(2) at 1-minute intervals to simulate the intrauterine pressures during the second stage of labor. Bloxsom developed the AL device in response to his hypothesis that the contractions of labor help to "condition: the infant for extrauterine survival. Parmalee said that the AL "certainly locks the infant up, safe from meddlesome and unintelligent treatment." When clear plastic versions of the AL became commercially available, it received widespread use in delivery rooms and newborn nurseries throughout the United States. In 1953, Apgar and Kreiselman produced apnea in adult dogs using pentobarbital and a muscle relaxant, and found that the AL device was unsuccessful with the oxygenation and ventilation of the animals. In 1954, Townsend in Rochester, New York, reported on his experience with the AL in 150 premature infants. He concluded that the AL should be "more accurately referred to as an oxygenator" and that, "the truly apneic infant cannot be maintained in a acyanotic state by the AL." The AL was finally subjected to the scrutiny of a randomized, controlled clinical trial that was published in 1956. Reichelderfer and Nitowski at Johns Hopkins randomized 171 infants to receive care in the AL or in an Isolette. Routine resuscitation, including positive pressure ventilation, was administered, as needed, to both study groups before placement into the AL or Isolette (Air Shields Inc, Hatboro, PA). They did not find any differences in the outcomes of the 2 study groups. By the mid 1950s, new information linking oxygen therapy and retrolental fibroplasia, led to a rapid decline in the use of the AL, even before the publication of the randomized trial. PMID- 11731644 TI - Annual summary of vital statistics: 2000. AB - The birth rate in 2000 (preliminary data) was 14.8 births per 1000 population, an increase of 2% from 1999 (14.5). The fertility rate, births per 1000 women aged 15 to 44 years, increased 3% to 67.6 in 2000, compared with 65.9 in 1999. The 2000 increases in births and the fertility rate were the third consecutive yearly increases, the largest in many years, halting the steady decline in the number of births and fertility rates in the 1990s. Fertility rates for total white, non Hispanic white, black, and Native American women each increased about 2% in 2000. The fertility rate for black women, which declined 19% from 1990 to 1996, has changed little since 1996. The rate for Hispanic women rose 4% in 2000 to reach the highest level since 1993. Birth rates for women 30 years or older continued to increase. The proportion of births to unmarried women remained about the same at one third, but the number of births rose 3%. The birth rate for teen mothers declined again for the ninth consecutive year. The use of timely prenatal care (83.2%) remained unchanged in 2000, and was essentially unchanged for non Hispanic white (88.5%), black (74.2%), and Hispanic (74.4%) mothers. The number and rate of multiple births continued their dramatic rise, but all of the increase was confined to twins; for the first time in more than a decade, the number of triplet and higher-order multiple births declined (4%) between 1998 and 1999 (multiple birth information is not available in preliminary 2000 data). The overall increases in multiple births account, in part, for the lack of improvement in the percentage of low birth weight (LBW) births. LBW remained at 7.6% in 2000. The infant mortality rate (IMR) dropped to 6.9 per 1000 live births (preliminary data) in 2000 (the rate was 7.1 in 1999). The ratio of the IMR among black infants to that for white infants was 2.5 in 2000, the same as in 1999. Racial differences in infant mortality remain a major public health concern. The role of low birth weight in infant mortality remains a major issue. Among all of the states, Utah and Maine had the lowest IMRs. State-by-state differences in IMR reflect racial composition, the percentage LBW, and birth weight-specific neonatal mortality rates for each state. The United States continues to rank poorly in international comparisons of infant mortality. Expectation of life at birth reached a record high of 76.9 years for all gender and race groups combined. Death rates in the United States continue to decline. The age-adjusted death rate for suicide declined 4% between 1999 and 2000; homicide declined 7%. Death rates for children 19 years of age or less declined for 3 of the 5 leading causes in 2000; cancer and suicide levels did not change for children as a group. A large proportion of childhood deaths, however, continue to occur as a result of preventable injuries. PMID- 11731645 TI - Does parental disapproval of smoking prevent adolescents from becoming established smokers? AB - OBJECTIVE: To evaluate the hypothesis that adolescents are less likely to smoke if their parents voice strong disapproval of smoking. DESIGN AND SETTING: Three wave school-based cohort study of rural Vermont adolescents attending 3 K-12 schools. We evaluate cross-sectional and longitudinal associations between perceived parental disapproval of smoking and the adoption of smoking behavior. OUTCOME MEASURES: Students' perceptions of their parents' reaction to their own smoking was ascertained by asking the following question for mothers and fathers: "How do you think your mother (father) would react if you were smoking cigarettes and she (he) knew about it?" A response of "S/he would tell me to stop and be very upset" was considered to indicate strong parental disapproval. Outcome measures include a 6-level smoking index for cross-sectional analyses and, for a longitudinal analysis of 372 never smokers at baseline, being an established smoker (smoked > or =100 cigarettes lifetime and within the past 30 days) by survey 3. RESULTS: The study samples for the cross-sectional analyses were 662 (baseline), 758 (year 2), and 730 (year 3). Students were equally distributed across grade (4th-11th grades) and gender. At baseline, most (65.9%) adolescents perceived both parents as disapproving of smoking, with 110 (16.6) perceiving 1 parent as disapproving, and 116 (17.5%) perceiving neither parent as disapproving. Perceived disapproval of smoking was inversely associated with adolescent smoking, grade in school, parental and sibling smoking, friend smoking, and ownership of tobacco promotional items. After controlling for confounding influences, adolescents who perceived strong parental disapproval of their smoking were less than half as likely to have higher smoking index levels compared with those who did not perceive strong parental disapproval. In the longitudinal sample of baseline never smokers, those who perceived strong disapproval in both parents at baseline were less than half as likely to become established smokers (adjusted odds ratio 0.4 [0.1, 1.0]). Those who perceived their parents becoming more lenient over time were significantly more likely to progress to established smokers. In all analyses, the effect of parental disapproval of smoking was stronger and more robust than the effect of parent smoking. In addition, the effect of parent disapproval was as strong for parents who smoked as it was for nonsmoking parents. An interaction analysis suggests that the peer smoking effect is attenuated when both parents strongly disapprove of smoking, suggesting that parent disapproval makes adolescents more resistant to the influence of peer smoking. CONCLUSIONS: These findings contrast with the widespread notion that there is little parents can do to prevent their adolescents from becoming smokers. Instead, adolescents who perceive that both parents would respond negatively and be upset by their smoking are less likely to smoke. Interventions that enhance parental self-efficacy in conveying and enforcing no-smoking policies for their children could reduce adolescent smoking. PMID- 11731646 TI - Comparison of continuity in a resident versus private practice. AB - BACKGROUND: There is widespread agreement among pediatric educators that continuity (following a panel of patients on a first contact basis for all their health care) is an important part of the education of pediatricians. OBJECTIVE: To measure continuity in a pediatric residency practice and to compare this continuity with 2 nearby private general pediatric group practices. We also examined measures of continuity suggested in the literature. DESIGN: Visit data were obtained from the practice billing system for a resident continuity practice and 2 nearby private practices for the 3-year period from July 1, 1992, to June 30, 1995. Visit data used were restricted to patients seen in the office of the practices during regular office hours. Continuity was measured using 5 different indices: 1) the usual provider of care index, visits by the usual clinician/total visits, 2) continuity for patient, the average proportion of visits that an individual patient was seen by his or her own physician, 3) continuity for physician (PHY), the average proportion of visits that an individual physician saw his or her own patients, 4) Continuity of Care Index (COC), and 5) the Modified, Modified Continuity Index. During the period examined, pediatric residents were present in the continuity practice for 1 half-day each week. The resident continuity practice (RCP) had 57 residents and saw 3386 patients for 18 955 visits. Private practice 1 (PP1) had 4 pediatricians who saw 4968 patients for 33 537 visits. Private practice 2 (PP2) had 5 pediatricians who saw 11 953 patients for 75 778 visits. RESULTS: For all visit types, continuity in the RCP was not as high as in the private practices, PHY-RCP versus PP1, PP2; 53% versus 70%, 77%. However, continuity in RCP was greater than 50% for all measures except the COC index, which precipitously decreases as the number of clinicians seen increases. Examining continuity for health maintenance visits (PHY-RCP, PP2 vs PP1; 96%, 96% vs 82%) RCP was equal to the best of the private practices. The percentage of patients not seen for a health maintenance visit during the study period was lowest in the resident practice (RCP/PP1/PP2, 15/22/30). CONCLUSIONS: Although continuity for all visits in this RCP was less than in private practice, it was surprisingly high, considering the limited time residents spend in clinic. In a particularly important area for continuity, health maintenance visits, continuity was identical to one and superior to the other private practice. PMID- 11731647 TI - Comparison of management strategies for extreme prematurity in New Jersey and the Netherlands: outcomes and resource expenditure. AB - OBJECTIVE: To quantify differences in resource expenditure in the perinatal period and long-term outcome of extremely premature infants who received systematically different approaches to neonatal intensive care. METHODS: Perinatal management, mortality, prevalence of disabling cerebral palsy (DCP), and resource expenditure of 2 population-based inception cohorts of extremely premature infants born in the mid-1980s were compared. Electronic fetal monitoring, tocolysis, cesarean section delivery, and assisted ventilation were used to characterize management approaches. Participants included all live births at 23 to 26 weeks' gestation in a 3-county area of central New Jersey (NJ) from 1984 to 1987 (N = 146) and throughout the Netherlands (NETH) in 1983 (N = 142). Mortality and the prevalence of DCP were the primary outcomes. Numbers of hospital days with and without assisted ventilation were the measures of resource expenditure. RESULTS: Electronic fetal monitoring (100% vs 38%), cesarean section (28% vs 6%), and assisted ventilation (95% vs 64%) were all more commonly used in NJ than in NETH. Ten percent of NJ deaths occurred without assisted ventilation, compared with 45% of Dutch deaths. A total of 1820 ventilator days were expended per 100 live births in NJ, compared with 448 in NETH. The increase in the number of nonventilator days (3174 vs 2265 days per 100 live births) did not reach statistical significance. Survival to age 2 (46 vs 22%) and the prevalence of DCP among survivors (17.2 vs 3.4%) were significantly greater in NJ at age 2 than in NETH at age 5. CONCLUSIONS: Near universal initiation of intensive care in NJ, compared with selective initiation of intensive care in NETH, was associated with 24.1 additional survivors per 100 live births, 7.2 additional cases of DCP per 100 live births, and a cost of 1372 additional ventilator days per 100 live births. PMID- 11731648 TI - C-reactive protein in febrile children 1 to 36 months of age with clinically undetectable serious bacterial infection. AB - OBJECTIVE: To determine the diagnostic properties of quantitative C-reactive protein (CRP) associated with clinically undetectable serious bacterial infection (SBI) in febrile children 1 to 36 months of age. METHODS: Febrile children presenting to a pediatric emergency department (ED) with ages ranging from 1 to 36 months, temperatures > or =39 degrees C, and clinically undetectable source of fever were enrolled in this prospective cohort study. Demographic information, ED temperature, duration of fever, and clinical evaluation using the Yale observation scale were recorded at the time of the initial evaluation. The white blood cell count (WBC), band count, absolute neutrophil count (ANC), and CRP concentration were measured at the same time. All patients received blood cultures and either a screening urinalysis or urine culture. A chest radiograph was obtained at the discretion of the ED physician. Patients with history of using antibiotics within 1 week of their presentation to the ED were excluded. The main outcome result was the presence of laboratory or radiographically proven SBI (bacteremia, meningitis, urinary tract infection, pneumonia, septic arthritis, and osteomyelitis). RESULTS: Seventy-seven patients were enrolled in the study. Fourteen (18%) had a SBI (6 urinary tract infection; 4 pneumonia, including 1 patient with Streptococcus pneumoniae bacteremia; and 4 occult S pneumoniae bacteremia), and 63 had no SBI. The 2 groups were indistinguishable in age, sex, ED temperature, duration of fever, and Yale observation scale. CRP concentration, WBC, and ANC were significantly different between the 2 groups. In a multivariate logistic regression analysis, only CRP remained as a predictor of SBI (Beta = 0.76, 95% confidence interval [CI]: 0.64, 0.89). Receiver-operating characteristic analysis demonstrated CRP (area under curve [AUC] 0.905, standard error [SE] 0.05, 95% CI: 0.808, 1.002) to be superior to ANC (AUC 0.805, SE 0.051, 95% CI: 0.705, 0.905) and to WBC (AUC 0.761, SE 0.068, 95% CI: 0.628, 0.895). A CRP cutoff point of 7 was determined to maximize both sensitivity and specificity (sensitivity 79%, specificity 91%, likelihood ratio 8.3, 95% CI: 3.8, 27.3). Multilevel likelihood ratios and posttest probabilities were calculated for a variety of CRP levels. A CRP concentration of <5 mg/dL effectively ruled out SBI (likelihood ratio 0.087, 95% CI: 0.02, 0.38, posttest probability of SBI 1.9%). CONCLUSIONS: Quantitative CRP concentration is a valuable laboratory test in the evaluation of febrile young children who are at risk for occult bacteremia and SBI, with a better predictive value than the WBC or ANC. PMID- 11731649 TI - Zinc supplementation in infants born small for gestational age reduces mortality: a prospective, randomized, controlled trial. AB - BACKGROUND: Low birth weight infants have been noted to have low zinc concentrations in cord blood, and zinc deficiency in childhood is associated with reduced immunocompetence and increased infectious disease morbidity. This study investigates whether zinc supplementation of infants born full term and small for gestational age affects mortality. METHODS: A randomized, double-blind, controlled trial with 2-by-2 factorial design enrolled 1154 full-term small for gestational age infants to receive in syrup 1 of the following: riboflavin; riboflavin and zinc (5 mg as sulfate); riboflavin, calcium, phosphorus, folate, and iron; or riboflavin, zinc, calcium, phosphorus, folate, and iron. A fixed dosage of 5 mL per child was given daily from 30 to 284 days of age. Household visits were made 6 days per week to provide the syrup and conduct surveillance for illness and death. When a child's death was reported, parental reports and medical records were used to ascertain the cause. The effects of zinc and of the combination of iron, folate, calcium, and phosphorus were analyzed by intent to treat. The mortality analysis was performed using a survival analytic approach that models time until death as the dependent variable; all models had 2 terms as independent variables: 1 for the zinc effect and 1 for the vitamin and mineral (calcium and phosphorus, folate and iron) effect. RESULTS: Zinc supplementation was associated with significantly lower mortality, with a rate ratio of 0.32 (95% confidence interval: 0.12-0.89). Calcium, phosphorus, folate, and iron supplementation was not associated with a mortality reduction, although a statistically nonsignificant trend toward reduction was observed with a rate ratio of 0.88 (95% confidence interval: 0.36-2.15). CONCLUSIONS: Zinc supplementation in small for gestational age infants can result in a substantial reduction in infectious disease mortality. PMID- 11731651 TI - Impact of early intervention on outcome after mild traumatic brain injury in children. AB - OBJECTIVE: The impact of mild head injury or mild traumatic brain injury (TBI) in children is variable, and determinants of outcome remain poorly understood. There have been no previous attempts to evaluate the impact of interventions to improve outcome. Results of previous intervention studies in adults have been mixed. This study aimed to evaluate the impact of providing information on outcome measured in terms of reported symptoms, cognitive performance, and psychological adjustment in children 3 months after injury. METHODS: A total of 61 children with mild TBI were assessed 1 week and 3 months after injury, and 58 children with mild TBI were assessed 3 months after injury only. They were compared with 2 control groups (N = 45 and 47) of children with minor injuries not involving the head. Participants completed measures of preinjury behavior and psychological adjustment, postconcussion symptoms, and tests of attention, speed of information processing, and memory. Children with mild TBI seen at 1 week were also given an information booklet outlining symptoms associated with mild TBI and suggested coping strategies. Those seen 3 months after injury only did not receive this booklet. RESULTS: Children with mild TBI reported more symptoms than controls at 1 week but demonstrated no impairment on neuropsychological measures. Initial symptoms had resolved for most children by 3 months after injury, but a small group of children who had previous head injury or a history of learning or behavioral difficulties reported ongoing problems. The group not seen at 1 week and not given the information booklet reported more symptoms overall and was more stressed 3 months after injury. CONCLUSIONS: Providing an information booklet reduces anxiety and thereby lowers the incidence of ongoing problems. PMID- 11731650 TI - Maternal and infant factors associated with failure to thrive in children with vertically transmitted human immunodeficiency virus-1 infection: the prospective, P2C2 human immunodeficiency virus multicenter study. AB - OBJECTIVE: Many children with human immunodeficiency virus-1 (HIV-1) have chronic problems with growth and nutrition, yet limited information is available to identify infected children at high risk for growth abnormalities. Using data from the prospective, multicenter P2C2 HIV study, we evaluated the relationships between maternal and infant clinical and laboratory factors and impaired growth in this cohort. METHODS: Children of HIV-1-infected women were enrolled prenatally or within the first 28 days of life. Failure to thrive (FTT) was defined as an age- and sex-adjusted weight z score < or =-2.0 SD. Maternal baseline covariates included age, race, illicit drug use, zidovudine use, CD4+ T cell count, and smoking. Infant baseline predictors included sex, race, CD4+ T cell count, Centers for Disease Control stage, HIV-1 RNA, antiretroviral therapy, pneumonia, heart rate, cytomegalovirus, and Epstein-Barr virus infection status. RESULTS: The study cohort included 92 HIV-1-infected and 439 uninfected children. Infected children had a lower mean gestational age, but birth weights, lengths, and head circumferences in the 2 groups were similar. Mothers of growth-delayed infants were more likely to have smoked tobacco and used illicit drugs during pregnancy. In repeated-measures analyses of weight and length or height z scores, the means of the HIV-1-infected group were significantly lower at 6 months of age (P <.001) and remained lower throughout the first 5 years of life. In a multivariable Cox regression analysis, FTT was associated with a history of pneumonia (relative risk [RR] = 8.78; 95% confidence interval [CI]: 3.59-21.44), maternal use of cocaine, crack, or heroin during pregnancy (RR = 3.17; 95% CI: 1.51-6.66), infant CD4+ T-cell count z score (RR = 2.13 per 1 SD decrease; 95% CI: 1.25-3.57), and any antiretroviral therapy by 3 months of age (RR = 2.77; 95% CI: 1.16-6.65). After adjustment for pneumonia and antiretroviral therapy, HIV-1 RNA load remained associated with FTT in the subset of children whose serum was available for viral load analysis. CONCLUSIONS: Clinical and laboratory factors associated with FTT among HIV-1-infected children include history of pneumonia, maternal illicit drug use during pregnancy, lower infant CD4+ T-cell count, exposure to antiretroviral therapy by 3 months of age (non-protease inhibitor), and HIV-1 RNA viral load. PMID- 11731652 TI - Cardiorespiratory events in preterm infants referred for apnea monitoring studies. AB - BACKGROUND: Episodes of apnea, desaturation, and bradycardia are common in preterm infants. Such infants who have persistent cardiorespiratory events detected by clinical bedside monitoring often are referred for overnight apnea monitoring studies. OBJECTIVE: To characterize apnea, bradycardia, and desaturation events in infants referred for an overnight apnea monitoring study and compare them with corresponding events in control infants of similar age and weight with no bedside monitor alarms. METHODS: Twelve-hour bedside apnea monitoring studies were performed on 68 preterm infants before hospital discharge. This population included 35 infants who were referred by their attending physicians because of persistent bedside monitor alarms (referral group) and 33 infants who had no documented cardiorespiratory events for at least 2 days before the study (control group). Each study monitored respiration via respiratory inductance plethysmography, oxygen saturation (Sao2), and heart rate. Events were defined as meeting 1 of the following criteria: apnea > or =20 seconds, bradycardia < or =80 beats per minute, or Sao2 < or =80%. RESULTS: The incidence of apnea > or =20 seconds was low, with no significant difference between infant groups. Referral infants exhibited a higher occurrence of desaturation episodes (20 +/- 6 vs 6 +/- 3 episodes/12-hour study) and a higher occurrence of bradycardia episodes (4.3 +/- 0.8 vs 1.1 +/- 0.3 episodes/12-hour study) than controls. These episodes of desaturation and bradycardia were always preceded by a respiratory pause, which was shorter in the referral infants (10.0 +/- 0.4 seconds vs 12.0 +/- 1.0 seconds). Baseline Sao2 was lower in referrals than controls (95 +/- 1% vs 98 +/- 1%), and the incidence of periodic breathing was significantly higher. CONCLUSIONS: Infants referred for apnea monitoring studies because of persistent bedside monitor alarms have very infrequent prolonged apnea but a higher frequency of desaturation and bradycardia in response to short respiratory pauses than infants without persistent bedside monitor alarms. Referral infants also exhibit a lower baseline Sao2. These abnormalities in oxygenation and cardiorespiratory control may be markers for subtle residual lung disease or functional central nervous system abnormalities. PMID- 11731653 TI - Intrauterine growth of full-term infants: impact of prenatal cocaine exposure. AB - OBJECTIVE: The objectives of this study were to estimate the effect of prenatal cocaine exposure on fetal growth and gestational age after controlling for exposure to alcohol, tobacco, and marijuana and other covariates; to evaluate whether prenatal cocaine exposure has a disproportionate adverse effect on head circumference compared with overall somatic growth; and to assess whether the effect of prenatal cocaine exposure on fetal growth is mediated by cocaine's suspected effect on gestational age. METHODS: The study population includes 476 neonates participating in the Miami Prenatal Cocaine Study, a longitudinal follow up of in utero cocaine exposure. The sample, restricted to full-term neonates born to African-American inner-city mothers, included 253 infants exposed prenatally to cocaine (with or without alcohol, tobacco, or marijuana exposure) and 223 non-cocaine-exposed infants, of whom 147 were drug-free and 76 were exposed to varying combinations of alcohol, tobacco, or marijuana. RESULTS: Evidence based on structural equations and multiple regression models supports a hypothesis of cocaine-associated fetal growth deficits (0.63 standard deviation) and an independent mild effect on gestational age (0.33 standard deviation). There was no evidence of a disproportionate adverse effect on birth head circumference once the impact on overall growth was estimated. There was evidence that some but not all of the cocaine effect on fetal growth was direct and some was indirect, acting via an intermediate influence of cocaine on gestational age. CONCLUSIONS: Cocaine-associated growth deficits, symmetrical and partially mediated by gestational age, were observed in this sample of inner-city African American full-term infants prospectively enrolled at birth. Long-term implications will be the subject of future reports from this longitudinal investigation. PMID- 11731654 TI - Newborn screening for congenital adrenal hyperplasia in the Netherlands. AB - OBJECTIVE: To evaluate whether congenital adrenal hyperplasia (CAH) patients can be detected by newborn screening before the occurrence of life-threatening salt wasting and whether the prevalence, specificity, and sensitivity are adequate enough for a routine screening procedure. DESIGN: From 1998, a 2-year regional pilot screening for CAH was performed. In 1998, cutoff levels for 17OHP were primarily based on birth weight, and in 1999 on gestational age. In addition, nationwide, all newly diagnosed patients with CAH were reported to the Dutch Pediatric Surveillance Unit to compare screened CAH patients with CAH patients in the area without screening. RESULTS: In 2 years, 176 684 newborns were screened and 15 CAH patients (7 males/8 females) were detected. Therapy was started at the median age of 7 days. In the area without screening, 223 307 infants were born and 19 CAH patients (10 males/9 females) were reported to the Dutch Pediatric Surveillance Unit. Therapy was started at the median age of 14 days. The mean (standard deviation) serum sodium concentration was 134.5 (3.4) mmol/L in the area of screening versus 124.5 (10.8) mmol/L in the area without screening. The overall prevalence was 1:11 764. In 1998 and 1999, the specificity was 99.76% and 99.97%, respectively. The positive predictive value was 4.5% and 16%, respectively. To date, no false-negative cases have been detected. CONCLUSION: Severe salt wasting can be prevented by neonatal screening. The prevalence, specificity, and sensitivity allowed addition of screening for CAH to the routinely performed national neonatal screening program. PMID- 11731655 TI - Functional limitations in young children with congenital heart defects after cardiac surgery. AB - With the recent dramatic decline in mortality rates of infants undergoing open heart surgery (OHS), there is growing concern regarding neurodevelopmental sequelae. Outcome studies have primarily focused on delineating developmental impairments; however, the impact on function and family burden has not been investigated. The objective of this study was to determine the prevalence of functional limitations and burden of care of young children with congenital heart defects (CHD) after OHS. STUDY DESIGN: One hundred thirty-one eligible infants with CHD undergoing their first OHS were recruited prospectively. Patients were assessed pre- and postoperatively, and again 12 to 18 months after surgery. Functional assessments included the WeeFIM (Functional Independence Measure) and the Vineland Adaptive Behavior Scale. RESULTS: For the WeeFIM, mean quotients were 84.3 +/- 23.8 (self-care), 77.2 +/- 30.0 (mobility), and 92.4 +/- 27.8 (cognition), with an overall quotient of 83.8 +/- 23.4. Only 21% of the cohort was functioning within their expected age range. Moderate disability was noted in 37%, while only 6% demonstrated a severe disability. For the Vineland scale, mean score for daily living skills was 84.4 +/- 17.6, and 80.3 +/- 15.9 for socialization. Functional difficulties in daily living skills were documented in 40%, whereas >1/2 had poor socialization skills. Factors enhancing risk for functional disabilities included perioperative neurodevelopmental status, microcephaly, length of deep hypothermic circulatory arrest, length of stay in the intensive care unit, age at surgery, and maternal education. CONCLUSIONS: The high prevalence of functional limitations and dependence in activities of daily living is currently underappreciated in the clinical setting, and deserves additional attention by pediatricians and developmental specialists. PMID- 11731656 TI - Patient self-management of acute asthma: adherence to national guidelines a decade later. AB - BACKGROUND AND OBJECTIVES: Children in the emergency department (ED) with acute asthma were enrolled to assess the impact of asthma on their activities of daily living and evaluate their access to care and preventive strategies, determine the proportion who adhered to the National Heart, Lung, and Blood Institute (NHLBI) guidelines for proper steps to take at home during an acute asthma exacerbation, and compare adherence rates for those with persistent and mild intermittent asthma. DESIGN AND METHODS: Children 2 to 18 years old who presented to the Children's Hospital of Philadelphia's ED with acute asthma exacerbations were enrolled prospectively. Parents and patients completed the 108-item Asthma Exacerbation Response Questionnaire with a focus on determining the home management steps they took both at the onset of the asthma exacerbation and just before coming to the ED. RESULTS: Among the 433 children studied, 76% had at least 1 doctor visit, 75% had at least 1 ED visit, and 43% had at least 1 hospitalization for asthma in the preceding 12 months. Overall, 64% had persistent asthma by NHLBI criteria, yet just 4% were cared for by an allergist or pulmonologist, 38% took daily anti-inflammatory therapy, and 18% received a daily inhaled corticosteroid. Also, 48% did not use a holding chamber with their metered-dose inhalers, and 66% did not use their peak flow meters. Regarding exacerbation response, 71% did not have a written action plan, and 89% did not maintain a symptom diary. Both at the onset of wheezing and just before coming to the ED, administration of a beta2-agonist was the only step that the majority of children performed. One-third or fewer followed the other steps recommended by the NHLBI, including using a peak flow meter, beginning oral corticosteroids, calling or going to see the doctor, or going to the ED. Children with persistent asthma were not more adherent to the guidelines than those with mild intermittent disease. CONCLUSIONS: Asthma has a significant adverse effect on the lives of these children. The NHLBI guidelines, first published a decade ago, were designed to reduce asthma's increasing morbidity and mortality, but this study uncovered a high rate of nonadherence with many aspects of the guidelines, including preventive strategies and home management of an exacerbation. PMID- 11731657 TI - Neurobehavioral deficits in premature graduates of intensive care--potential medical and neonatal environmental risk factors. AB - There is growing evidence that a large number of very low birth weight infants are exhibiting neurobehavioral problems in the absence of cerebral palsy at follow-up that has extended into school age and adolescence. Many clinical factors (ie, chronic lung disease, recurrent apnea and bradycardia, transient hypothyroxemia of prematurity, hyperbilirubinemia, nutritional deficiencies, glucocorticoid exposure), as well as stressful environmental conditions, including infant-provider interaction, constant noise, and bright light, may act in combination to impact on the developing brain, even in the absence of overt hemorrhage and/or ischemia. Any potential intervention strategy designed to prevent cognitive and behavioral problems has to account for the numerous biological and clinical conditions and/or interventions, as well as postdischarge social and environmental influences. PMID- 11731658 TI - Joseph W. St. Geme, Jr address 2001: can one have a successful academic career in 2001? PMID- 11731659 TI - Quality in pediatrics: a progress report for the American Academy of Pediatrics. PMID- 11731660 TI - Targeted screening for elevated blood lead levels: populations at high risk. PMID- 11731661 TI - Zinc supplementation saves the lives of children living in poverty. PMID- 11731662 TI - Management of tick bites and early Lyme disease: a survey of Connecticut physicians. AB - Practice guidelines have been published for the treatment of Lyme disease (LD). These guidelines have been challenged as inadequate. Two common LD management problems are antibiotic prophylaxis of deer tick bites (deer ticks may carry Borrelia burgdorferi, the spirochete that causes LD) and antibiotic treatment of erythema migrans, the pathognomonic rash of LD. A 1-page questionnaire was sent to a 13% (573/4300) sample of Connecticut physicians to define how they treat deer tick bites and erythema migrans. Questionnaires were returned by 320 (56%) of 573 physicians. Questionnaires were analyzed for the 267 physicians who saw patients with LD. Seventy (26%) of the 267 surveyed physicians prescribed antibiotic prophylaxis for patients with tick bites. B burgdorferi serology was ordered by 31% of physicians for patients with tick bites. Most surveyed physicians treated erythema migrans with doxycycline or amoxicillin for a mean of 21 days. Serology was ordered by 49% of physicians for patients with erythema migrans. Most physicians did not use prophylaxis for patients with deer tick bites. In addition, most of the physicians surveyed followed established guidelines for treating patients with erythema migrans. However, many of the physicians surveyed do serologic testing for patients with tick bites and/or erythema migrans. Serologic testing for these patients is usually not necessary. PMID- 11731663 TI - Fatty acids, alpha-fetoprotein, and cystic fibrosis. AB - Docosahexaenoic acid, found lacking in animal models of cystic fibrosis, has been proposed as a dietary supplement therapy for this genetic disorder. Alpha fetoprotein, which binds and transports docosahexaenoic acid, may be a useful marker to improve the management and follow-up in newborn screening programs for cystic fibrosis, because only 20% of such infants are diagnosed at birth. PMID- 11731664 TI - Sn-Mesoporphyrin interdiction of severe hyperbilirubinemia in Jehovah's Witness newborns as an alternative to exchange transfusion. AB - OBJECTIVE: The religious convictions of parents who are Jehovah's Witness adherents lead them to reject the use of exchange transfusions as therapy for severe hyperbilirubinemia in newborns in whom intensive phototherapy has failed to control this problem. Consequently, physicians caring for such infants may be obliged to initiate legal action to compel use of the procedure when severe hyperbilirubinemia not sufficiently responsive to phototherapy warrants an exchange transfusion. Our goal was to determine if we could use the potent inhibitor of bilirubin production, Sn-Mesoporphyrin (SnMP), to resolve the troubling medical-legal issues in such situations in 2 infants with hemolytic disease of the newborn who required exchange transfusions for severe hyperbilirubinemia but whose Jehovah's Witness parents rejected the procedure. SnMP was administered in a single dose, as in previous studies, at the time when exchange transfusion would have been initiated and plasma bilirubin levels were monitored at close intervals thereafter. METHODS: SnMP is a potent inhibitor of heme oxygenase, the rate-limiting enzyme in catabolism of heme to bilirubin. We found in earlier studies that in single doses of 6 micromol/kg birth weight, SnMP is extremely effective in moderating the course of hyperbilirubinemia and in eliminating the need for supplemental phototherapy in jaundiced newborns. In the 2 cases described, a single dose of SnMP (6 micromol/kg birth weight) was administered intramuscularly to severely jaundiced infants with immune hemolysis at a time when clinical circumstances dictated the need for exchange transfusion. CASE 1: This patient was a preterm male infant (gestational age: 35 5/7 weeks; birth weight: 2790 g) whose plasma bilirubin concentration (PBC) at 1 hour after birth was 5.0 mg/dL. Despite intensive phototherapy with 3 banks of lights and 1 biliblanket, the PBC increased steadily with no diminution in the rate of increase for 75 hours. In view of the problems of immune hemolysis, and prematurity, and the inability of phototherapy to stop progression of hyperbilirubinemia, a decision to carry out an exchange transfusion was made; the decision was, however, rejected by the Jehovah's Witness parents. Pending legal action to compel use of the procedure, a request to this (Rockefeller) laboratory for SnMP was made; its use was approved by the Food and Drug Administration; and the inhibitor was delivered to the physician-in-charge (D.P.M.) in Sioux Falls, South Dakota. The single dose of SnMP was administered to the infant at 75 hours after birth; the course of hyperbilirubinemia before and after the use of the inhibitor is shown in Fig 1. [figure: see text]. CASE 2: This female term infant (gestational age: 38-39 weeks; birth weight: 4140 g) with immune hemolysis was delivered by cesarean section and because of problems related to meconium aspiration required helicopter transfer to the Special Care Nursery in Abilene, Texas, where 10 hours after birth the first PBC was determined to be 18.0 mg/dL. Double-bank phototherapy plus a biliblanket was initiated; a third bank of lights was later ordered. The PBC fluctuated in the ensuing 2 days between 13.8 to 25.8 mg/dL during which suggestive clinical signs of possible bilirubin encephalopathy became manifest. In view of the clinical circumstances and the continued severe hyperbilirubinemia, permission for a double-exchange transfusion was requested. The parents, who were Jehovah's Witness adherents, refused the procedure. While preparing legal action to compel use of the exchange, a request was made to this (Rockefeller) laboratory for use of SnMP to attempt control of hyperbilirubinemia. With FDA approval, the SnMP was delivered to the attending neonatologist (J. R. M.) in Abilene and administered in a single dose (6 micromol/kg birth weight) at 56 hours after birth when the PBC was 19.5 mg/dL. The course of bilirubinemia before and after SnMP use is shown in Fig 2. [figure: see text]. RESULTS AND CONCLUSIONS: The use of SnMP to moderate or prevent the development of severe hyperbilirubinemia in newborns (preterm, near-term, term with high PBCs [15-18 mg/dL], ABO-incompatibility; glucose-6-phosphate dehydrogenase deficiency) has been extensively studied in carefully conducted clinical trials the results of which have been reported earlier. This inhibitor of bilirubin production has demonstrated marked efficacy in moderating the course of hyperbilirubinemia in all diagnostic groups of unconjugated neonatal jaundice. The 2 cases described in this report confirmed the efficacy of SnMP in terminating progression of hyperbilirubinemia in infants in whom phototherapy had failed to sufficiently control the problem and whose parents, for religious reasons, would not permit exchange transfusions. Interdiction of severe hyperbilirubinemia by inhibiting the production of bilirubin with SnMP can be an effective alternative to the use of exchange transfusion in the management of severe newborn jaundice that has not responded sufficiently to light treatment to ease concern about the development of bilirubin encephalopathy. PMID- 11731665 TI - Guiding principles, attributes, and process to review medical management guidelines. AB - Few issues are more central to the ongoing debate about health care in the United States than concerns about cost and quality of medical care. The recent development and implementation of medical management guidelines that include recommendations for diagnostic and therapeutic interventions, hospital length of stay, intensity of service, home care, and access to specialists have often focused this debate on the potential trade-off between cost reductions and quality of care. The American Academy of Pediatrics recognizes that cost and quality are integrally related and that it is possible to reduce costs while maintaining and improving quality. The purpose of this statement is to help pediatricians and other health care providers interpret, evaluate, and improve medical management guidelines. PMID- 11731666 TI - Continuous positive airway pressure: randomized, controlled trial in Australia. PMID- 11731667 TI - Caution about trial methodology and statistics. PMID- 11731668 TI - Health supervision for children with Down syndrome. PMID- 11731669 TI - Screening for retinopathy of prematurity. PMID- 11731670 TI - Antibiotics and sinusitis. PMID- 11731671 TI - Disturbed by questionnaire. PMID- 11731672 TI - Clinical auscultation skills in pediatric residents. PMID- 11731673 TI - Congenital rubella infection control problem. PMID- 11731674 TI - Iron supplementation during erythropoietin treatment. PMID- 11731675 TI - The AAP and gun control. American Academy of Pediatrics. PMID- 11731676 TI - Prevention of hepatitis B in adolescents. PMID- 11731677 TI - The FOPE II commentary. Future of Pediatric Education II. PMID- 11731678 TI - Head lice infestation: single drug versus combination therapy. PMID- 11731679 TI - Nocturnal enuresis: current concepts. PMID- 11731681 TI - Visual diagnosis: a 10-week-old infant who has jaundice. PMID- 11731682 TI - Index of suspicion. PMID- 11731683 TI - The immigrant, refugee, or internationally adopted child. PMID- 11731684 TI - Pre-operative intra-aortic balloon counterpulsation in stable patients with left main coronary disease. AB - OBJECTIVE: To determine whether prophylactic, pre-operative, intra-aortic balloon counterpulsation (IABC) improves clinical outcome in stable patients with severe left main coronary artery disease. METHODS: A post-hoc analysis of 457 prospectively tracked, non-randomized patients undergoing coronary artery bypass graft surgery (CABG) for left main stenoses 50% and multivessel coronary disease, but without any hemodynamic compromise or ongoing angina, was conducted. Patients with heart failure, shock, ongoing ischemia or previous CABG were excluded. In 287 patients, pre-operative IABC was not used (Group 1), while IABC was initiated in 170 patients for "prophylaxis" (Group 2). RESULTS: Groups 1 and 2 were similar in age (67 +/- 10 years versus 67 +/- 11 years, respectively), sex (72% male versus 71% male, respectively), and body mass index (28 +/- 5.5 versus 27 +/- 5.1, respectively). However, more Group 1 patients had peripheral vascular disease (PVD) (25% versus 11%), but more Group 2 patients had diabetes (37% versus 29%), and a lower left ventricular ejection fraction. The unadjusted 30 day mortality was significantly higher in Group 1 [16 (5.6%) versus 2 (1.2%); p = 0.02]. Cardiopulmonary bypass time and post-operative length of stay did not differ between the two groups. After adjusting for PVD in the multivariate analysis, the p-value for the no IABP versus IABP comparison in the presence of PVD was 0.10, even though 0/18 patients with PVD and IABC died. CONCLUSION: While unadjusted mortality appears lower with prophylactic IABC, confounding variables such as PVD mandate a larger, randomized clinical trial in order to establish the role of IABC in stable patients with left main disease. PMID- 11731685 TI - Dual antiplatelet therapy with clopidogrel and aspirin after carotid artery stenting. AB - BACKGROUND: Carotid artery stenting is being used as an alternative to carotid endarterectomy, both within the context of clinical trials and in non-surgical candidates. Though stenting is known to activate platelets, the role of antithrombotic therapy in carotid stenting has not been fully characterized. METHODS AND RESULTS: Consecutive patients (n = 162) were followed in a single center carotid stent registry. The cumulative rate of 30-day death, stroke, transient ischemic attack and myocardial infarction in those patients receiving a thienopyridine was determined, as were rates of stent thrombosis and intracranial hemorrhage. The mean age of the patients was 70.3 years and there was an extremely high prevalence of cardiovascular comorbidities, including 40% with unstable angina. The carotid lesion was symptomatic in 59% of patients. The average pre-treatment stenosis was 83%. The cumulative 30-day rate of death, stroke, transient ischemic attack and myocardial infarction was 5.6%. Specifically, in the patients who received ticlopidine (n = 23), the rate was 13%, versus 4.3% in the patients who received clopidogrel (n = 139) (p = 0.01). In this series, there were no cases of stent thrombosis and 1 intracranial hemorrhage. CONCLUSION: Dual antiplatelet therapy with clopidogrel plus aspirin in patients receiving carotid artery stents is associated with a low rate of ischemic events. Furthermore, clopidogrel appears superior to ticlopidine. Thus, our findings lend support to the dual antiplatelet strategy of clopidogrel plus aspirin for patients undergoing carotid artery stenting. PMID- 11731686 TI - Aspirin plus clopidogrel for everyone: panacea comes true in the 21st century. PMID- 11731687 TI - Controlled balloon inflation reduces long-term restenosis after percutaneous transluminal coronary angioplasty. AB - PURPOSE: The trauma induced by balloon angioplasty has an impact on the outcome of coronary interventions, such as stent procedures. However, balloon inflation for PTCA is not yet standardized even though procedural and long-term outcomes might be affected. METHODS: During routine PTCA, a total of 454 patients [mean age, 60.9 +/- 9.0 years; 162 (35.7%) with 1-vessel disease; 159 (35%) with 2 vessel disease; 133 (29.3%) with 3-vessel disease] were allotted to computer assisted dilatation (CAPS) with a pressure slope of 0.2 bar/s (CAPS 0.2; n = 149 patients), 1.0 bar/s (CAPS 1.0; n = 154 patients) or to standard inflation with a hand-driven pump (n = 151 patients). Angiographic follow-up rates after 4.1 +/- 3.2 months were 88.1% for the hand-driven pump, 94% for CAPS 0.2 and 87.7% for CAPS 1.0. RESULTS: Flow reducing (1.3-2.0%) and non-flow reducing (12.6-14.9%) dissections were equally distributed among all groups as were major adverse cardiac events (2.6-4.0%). The stent rate was 1.3% with the hand-driven pump, 0.7% with CAPS 0.2 and 1.3% with CAPS 1.0. Angiographic restenosis rate was 48.9% with the hand-driven pump, 44.3% with CAPS 0.2 and 32.6% with CAPS 1.0. (hand driven pump versus CAPS 1.0, p < 0.007; CAPS 0.2 versus CAPS 1.0, p < 0.049). CONCLUSIONS: The pressure slope during balloon inflation in PTCA has a significant impact on restenosis. The impact on stent procedures has yet to be determined. PMID- 11731688 TI - Treatment of long, diffuse, in-stent restenotic lesions with beta radiation using strontium 90 and sequential positioning "pullback" technique: procedural details and clinical outcomes. AB - BACKGROUND: Long, diffuse, in-stent restenotic lesions have been problematic for many patients, necessitating the need for multiple repeat percutaneous coronary interventions (PCI). The need for repeat PCI has been significantly reduced in patients since the advent of vascular brachytherapy. In-stent restenosis resulting in long lesions (> 30 mm) presents even more of a challenge. The interventional injury length created during PCI is usually greater than 30 mm and the vascular brachytherapy systems available in most hospitals are 30 mm or 40 mm in length. The purpose of this paper is to define "pullback technique" and to report the Montreal Heart Institute (MHI) data to show that the pullback technique using the Novoste system seems to be a safe and effective method for using vascular brachytherapy to treat long, diffuse, in-stent restenotic lesions. METHODS: We reviewed the database of patients enrolled in the Compassionate Use Registry between August 1999 and July 2000. The data are reported on 23 consecutive patients treated with the pullback technique. RESULTS: The mean lesion length was 49.4 +/- 19.8 mm. Three patients (13%) underwent target vessel revascularization. Angiographic follow-up was obtained in 18 patients (78%). Seven patients (38.8%) showed angiographic restenosis (> 50% luminal re narrowing). No aneurysms or zones of ectasia were noted. CONCLUSION: This angiographic and clinical evaluation of the MHI Compassionate Use Registry data show the pullback technique to be safe from both a dosimetric and clinical point of view. PMID- 11731689 TI - Reducing bedrest following arterial puncture for coronary interventional procedures--impact on vascular complications: the BAC Trial. AB - BACKGROUND: The optimal time for ambulating patients following femoral arterial puncture for percutaneous coronary intervention is unknown. This study was designed to test the hypothesis that reducing time to ambulation from 6 hours to 2 hours would not increase vascular complications. METHODS: In this randomized, controlled, single-center trial, patients were randomly assigned to receive either 2, 4 or 6 hours (control group) of bedrest after hemostasis was achieved at the femoral arterial puncture site. A total of 354 patients, enrolled from March 1997 to October 1998, participated in the study with 299 completing the full protocol. The primary endpoint was the incidence of vascular complications and any resulting interventions or outcomes including surgery, blood transfusion, prolonged length of stay or ultrasound compression. RESULTS: The 2 hour and 4 hour experimental groups were comprised of 99 patients each, while the 6 hour control group was comprised of 101 patients. There were no differences in vascular complications between the groups. A subgroup analysis of patients who received abciximab (n = 43) did not reveal any differences between groups. Only one patient developed a pseudoaneurysm (p = 0.36), requiring surgical repair and a prolonged length of stay. CONCLUSIONS: Patients undergoing femoral arterial puncture for coronary interventional procedures can safely ambulate 2 hours after hemostasis of the puncture site. Patients receiving abciximab as part of their coronary procedure may be able to ambulate as early as 2 hours following hemostasis. PMID- 11731690 TI - Early mobilization following percutaneous coronary intervention. PMID- 11731691 TI - Percutaneous mitral balloon valvotomy: six-year follow-up. AB - OBJECTIVE: Percutaneous mitral valvotomy (PMV) is an alternative to the surgical treatment of mitral stenosis. Results obtained with PMV appear to depend on the echocardiographical characteristics of the valvular apparatus. The purpose of this study was to report the immediate and late-term results with PMV. The incidence of late events (restenosis, mitral valve replacement and death), and their correlation with echocardiographic score (Wilkin's score) are also discussed. METHODS: Between December 1987 and August 1999, a total of 160 PMVs were performed at our institution. Ninety-six patients with a minimum of 6 months follow-up and echocardiographic evaluation of the mitral valve (Wilkin's score) before and after the procedure were selected for this study. Follow-up was available for 99% of the patients, with a mean follow-up of 33 +/- 22 months (range, 6 months to 11 years). Hazard ratio (HR) and Cox's regression were used for statistical analyses. RESULTS: PMV was successfully performed in 97% of the cases; in 84%, the result was considered optimal. The incidence of complications related to the procedure was 10%; no mortality was observed due to PMV. Severe mitral regurgitation was observed in 7% of the patients, but only 3% of the total group developed ventricular dysfunction or worsened their New York Heart Association functional class. Eight-four percent of the patients were free of late events at the end of the follow-up period. A restenosis rate of 34% was observed during follow-up; this rate did not correlate with age, functional class or atrial fibrillation. Restenosis was associated with pulmonary hypertension (HR 2.85; 95% confidence interval, 0.68-11.80). Also, Wilkin's score was not useful to predict the development of restenosis or clinical events in the mid- to long term. CONCLUSION: In our series, PMV had a high immediate success rate and a low incidence of complications due to the procedure. Incidence of late events was also low and was unrelated to the Wilkin's score; however, recurrence of stenosis was observed in one-third. Pulmonary hypertension should be considered to be an important clinical predictor of restenosis. PMID- 11731692 TI - Letting the air Out of the follow-up balloon. PMID- 11731693 TI - Initial clinical experience with implantable loop recorders. AB - The purpose of this study was to review the initial experience of a university hospital with Implantable Loop Recorders (ILR) for diagnosis of recurrent unexplained syncope or presyncope. Twelve patients with syncope or presyncope of unknown etiology (who had a negative tilt table test, electrophysiologic study, and neurologic work-up) underwent implantation of ILR. All implants were performed using the Reveal ILR (Medtronic AVE, Santa Rosa, California). The 8 cc device is 61 mm long, 19 mm wide, 8 mm thick and weighs 17 grams. It has 18 months of battery life and has 2 electrodes with 38.5 mm spacing. The device is nonvascular and is implanted approximately 2 fingerbreadths below the clavicle in a subcutaneous pocket (1.5 inches long) and is secured via polydacron suture to the pre-pectoral fascia/pectoralis muscle. Twelve patients with a mean age of 61 +/- 22 years received the ILR. Ten patients had syncope and 2 had presyncope. Three patients had coronary artery disease and 2 had dilated cardiomyopathy. ILRs were implanted for a mean follow-up period of 7.2 +/- 5.8 months (range, 1 day to 18 months). Two patients still continue to be monitored at the time of this report. The mean number of events prior to ILR was 6.0 +/- 5.4. Eight patients (66%) had recurrent syncope after implantation. One patient was not available for follow-up. There were no significant complications from the implant. In 5/12 patients (42%), the ILR helped to diagnose the etiology of the syncopal episode. Syncope was secondary to asystole in three patients, junctional bradycardia in another patient, and seizure activity in a fifth patient (high-frequency noise recorded on the electrocardiogram during sinus rhythm). The 4 patients with ILR documented bradyarrhythmias underwent permanent pacemaker implantation and are alive and well. ILR implantation is a simple, useful and safe method in assisting with the diagnosis of recurrent unexplained syncope or presyncope after an inconclusive electrophysiologic and neurologic evaluation. PMID- 11731694 TI - Negative remodeling at the ostium of the left circumflex artery. AB - We report an ostial lesion with negative remodeling. Coronary angiography revealed a 60% stenosis at the ostium of the left circumflex artery (LCX). Intravascular ultrasound (IVUS)-guided directional atherectomy followed by stenting was planned. However, IVUS images revealed no significant stenosis and negative remodeling at the ostium of the LCX. The lesion did not undergo intervention. PMID- 11731695 TI - Percutaneous coronary intervention of the left main trunk in congenitally anomalous single coronary artery. AB - Anomalous origin of the coronary arteries is infrequent and a single coronary artery is seen even less frequently. Accordingly, few reports have described percutaneous coronary interventions in this anomaly. We report successful balloon angioplasty and stenting of a left main trunk originating from the right coronary artery. PMID- 11731696 TI - Residual inferior atrial septal defect after surgical repair: closure under intracardiac echocardiographic guidance. PMID- 11731697 TI - The STRO-1+ marrow cell population is multipotential. AB - Human marrow-derived STRO-1+ cells were tested for their ability to differentiate into multiple mesenchymal phenotypes. STRO-1+ cells were isolated from long-term cultures of human marrow cells by magnetic immunobeads linked to the antibody STRO-1. Stromal layers generated from STRO-1+ cells, when cocultured without exogenous cytokines for several weeks with CD34+/CD38+ and CD34+/CD38low cells, were able to induce a 10- to 20-fold increase in colony-forming units, and to sustain cobblestone area colony-forming cells with a distinct time course for the different hematopoietic precursors. A myofibroblastic and adipocytic phenotype was observed in hematopoiesis-supportive stromal cells, as shown by electron microscopy and staining of lipids with Nile red. The adipocytic potential of STRO 1+ cells was further demonstrated by flow cytometry of Nile red O-stained cells cultured in adipocyte-induced medium. The same population of STRO-1+ cells cultured in osteoblast-inducing medium was shown to have osteogenic potential, as demonstrated in vitro by the formation of mineral (von Kossa staining), and in vivo by the formation of bone tissue within porous calcium phosphate ceramics implanted into athymic mice. This population of STRO-1+ cells was also shown to have chondrogenic potential in aggregate cultures which displayed a morphologic appearance of cartilage and stained positively with antibodies specific to type II and type X collagens. These results show that the subset of marrow cells that express the STRO-1 antigen is capable of differentiating into multiple mesenchymal lineages including hematopoiesis-supportive stromal cells with a vascular smooth muscle-like phenotype, adipocytes, osteoblasts and chondrocytes. PMID- 11731698 TI - Embryonic submandibular gland morphogenesis: stage-specific protein localization of FGFs, BMPs, Pax6 and Pax9 in normal mice and abnormal SMG phenotypes in FgfR2 IIIc(+/Delta), BMP7(-/-) and Pax6(-/-) mice. AB - Embryonic submandibular salivary gland (SMG) initiation and branching morphogenesis are dependent on cell-cell communications between and within epithelium and mesenchyme. Such communications are typically mediated in other organs (teeth, lung, lacrimal glands) by growth factors in such a way as to translate autocrine, juxtacrine and paracrine signals into specific gene responses regulating cell division and histodifferentiation. Using Wnt1-Cre/R26R transgenic mice, we demonstrate that embryonic SMG mesenchyme is derived exclusively from cranial neural crest. This origin contrasts to that known for tooth mesenchyme, previously shown to be derived from both neural crest and nonneural crest cells. Thus, although both SMGs and teeth are mandibular derivatives, we can expect overlap and differences in the details of their early inductive interactions. In addition, since embryonic SMG branching morphogenesis is analogous to that seen in other branching organs, we also expect similarities of expression regarding those molecules known to be ubiquitous regulators of morphogenesis. In this study, we performed an analysis of the distribution of specific fibroblast growth factors (FGFs), FGF receptors, bone morphogenetic proteins (BMPs) and Pax transcription factors, previously shown to be important for tooth development and/or branching morphogenesis, from the time of initiation of embryonic SMG development until early branching morphogenesis. In addition, we report abnormal SMG phenotypes in FgfR2- IIIc(+/Delta), BMP7(-/-) and Pax6(-/-) mice. Our results, in comparison with functional studies in other systems, suggest that FGF-2/FGFR-1, FGF-8/FGFR-2(IIIc) and FGF-10/FGFR-2(IIIb) signaling have different paracrine and juxtacrine functions during SMG initial bud formation and branching. Finally, our observations of abnormal SMGs in BMP7(-/-) and Pax6(-/-) indicate that both BMP7 and Pax6 play important roles during embryonic SMG branching morphogenesis. PMID- 11731699 TI - Energy-dependent types of cell death in MCF-7 breast cancer cell tumors implanted into nude mice. AB - The present study aims at verifying whether aponecrosis, a novel type of cell demise sharing the features of both apoptosis and necrosis previously identified in in vitro experiments, could represent a natural phenomenon that also occurs in vivo. The pathways of cell death were analyzed morphologically in MCF-7 breast cancer cell tumors implanted into nude mice. These tumors grew as encapsulated masses devoid of blood vessels, thus allowing for the creation of a gradient of oxygen and nutrients from the periphery inwards. Expression of wild-type p53 and DNA fragmentation were investigated as apoptotic markers. Further experiments were carried out on MCF-7 cells cultured in vitro in the presence and absence of oxygen and nutrients. In these cultures, the intracellular ATP levels were measured and related to the types of cell death recognized morphologically. Apoptotic cells were observed in the outer portion of the MCF-7 cell tumors close to blood vessels. In the inner portion of the tumors which lacks blood supply, several cells simultaneously showed signs of both apoptosis and necrosis, being identifiable as aponecrotic cells. Similarly, aponecrotic cells were also observed in the starved cultures, in concomitance with intracellular ATP depletion. These findings suggest that MCF-7 cells may accomplish the apoptotic process only when sufficient oxygen and energy supply are available. As the energy level decreases, aponecrosis may ensue as the result of an incomplete execution of the apoptotic program. PMID- 11731700 TI - Ultrastructural changes in the uterine luminal and glandular epithelium during the oestrous cycle of the marsupial Monodelphis domestica (grey short-tailed opossum). AB - Ultrastructural changes in the endometrium associated with the oestrous cycle were studied in the South American marsupial Monodelphis domestica. The most conspicuous changes include the height and the differentiation of the uterine luminal and glandular epithelium, which consists of ciliated and non-ciliated cells. The glandular epithelium attains its maximum development during oestrus, the luminal epithelium at postoestrus. A distinct increase in the number of ciliated cells can be observed during pro-oestrus, reaching a maximum number at oestrus; this is followed by a process of deciliation. The presence of solitary cilia on the apices of non-ciliated cells is very conspicuous during all oestrous stages and can best be seen on the luminal epithelium. These findings differ from the observations in eutherian mammals, where solitary cilia are only found in the immature uterus or after ovariectomy. The secretory activity of non-ciliated cells of the luminal epithelium is hardly noticeable along the apical membrane and stains only very faintly with Alcian blue. The glandular epithelium cells are filled apically with exocytotic vesicles at oestrus and early postoestrus. However, in contrast to the cervical gland cells, they hardly stain with Alcian blue, indicating that mucins of a different type must be present. Mechanisms for the remodelling of the luminal and glandular epithelium are especially conspicuous at metoestrus and early pro-oestrus and include the presence of autolysosomes, residual bodies and apoptotic bodies. In the endometrial stroma, around the uterine glands, macrophages accumulate and attain a typical oestrous stage-dependent appearance during their phagocytotic activities. PMID- 11731701 TI - Sieve plugs in fenestrae of glomerular capillaries--site of the filtration barrier? AB - The exact location of the filtration barrier of the glomerular capillary wall, which consists of an endothelium, a basement membrane and a visceral epithelium, has not yet been determined. Apparent discrepancies between different investigators in the past could be explained if postmortem artifactual tissue changes, due to subnormal blood pressure or anoxia, have taken place in the endothelium before the tissue and tracers have been sufficiently fixed and immobilized by the fixative. To test this supposition, a new method of fixation, which includes a technique to maintain a physiological perfusion pressure and a fixative composed of an oxygen-carrying blood substitute fluid containing glutaraldehyde, was employed combined with contrast enhancement. New observations of the glomerular capillary wall revealed that filamentous plugs (about 90 nm in height) filled the capillary fenestrae and a filamentous surface coat about 60 nm thick covered the interfenestral domains of the endothelial cell. Based on these purely morphological data, we dare to suggest that the fenestral plugs are the primary site of the glomerular filtration barrier - albeit highly speculative, nevertheless a logical location - and consequently that the glomerular filtration process is a 'tangential-flow' as opposed to a 'dead-end' filtration process. A tangential-flow filtration would minimize 'clogging' and 'concentration polarization' in the 'filter'. PMID- 11731702 TI - Ultrastructural changes of the neuromuscular junction in reperfusion injury. AB - Ischemia followed by reperfusion of skeletal muscle frequently takes place in trauma surgery. Anoxia followed by reoxygenation leads to reperfusion injury, which damages the involved tissues. However, no information is available about how the neuromuscular junction is affected by ischemia-reperfusion. Tourniquet ischemia of the left hind limb was applied in the anesthetized rat for 2 h. Reperfusion lasted for 2 and 24 h and for 1, 2 and 4 weeks. The extensor digitorum longus and the soleus muscles from both legs were prepared for electron microscopic analyses. Morphological changes of the neuromuscular junction were investigated. In all cases only the nerve endings (terminals) were affected. The postsynaptic structures were not affected. Changes can be grouped in two categories: degeneration and recovery. Degeneration consisted of the loss of synaptic vesicles, disruption of the presynaptic membrane, degeneration of the mitochondria and the development of vacuoles. It was most severe at 24 h and was still present at 4 weeks of reperfusion. Recovery started at 1 week of reperfusion and lasted at least for 4 weeks. It consisted of the slow reappearance of synaptic vesicles and mitochondria, and restoration of the presynaptic membrane with active zones. Ultrastructure of the skeletal muscle fibers did not show pathological changes. The recovery of the structures may be regulated by the Schwann cells and also by the postsynaptic membrane which is not affected by 2 h of ischemia followed by reperfusion. PMID- 11731703 TI - Sphincteric musculature of female canine urethra in comparison to woman including 3D reconstruction. AB - The circular arranged sphincteric musculature of bladder neck of female dogs and women were studied histomorphologically. 3D reconstructions of the anatomy of the bladder neck improve the understanding of construction principles of the musculature of the lower urinary tract and help to compare both species. Our own investigations based on 12 adult female canine and 15 female human autopsy preparations. The special feature of our study was the extensive en bloc preparation of all the organs of the lower urinary tract and surrounding organs. The organ blocks were reprocessed in complete serial sections from the bladder outlet down to the bulb of vestibuli. Despite different detailed construction, in both species the striated sphincter musculature of the urethra is an independent morphological unit. There is no continuation of pelvic floor muscles to the urethra. In humans, the urethral sphincter consists of a smooth muscular part (m. sphincter urethrae glaber) and a striated part (m. sphincter urethrae transversostriatus). In the female dog, striated muscle fibres encircle the urethra in the middle third exclusively. In the distal third of the urethra, it encircles the urethra and the vagina. In the female dog, the lamellae of detrusor continue directly to the urethra. Throughout the cranial and middle third of the urethra, smooth muscle cell bundles form a homogenous compact sphincteric muscle originating from the middle circular layer of detrusor. In that way, no true bladder neck sphincter according to the m. sphincter vesicae in women exists in dogs. According to the smooth muscular part of the m. sphincter urethrae in women, for this musculature the term m. sphincter urethrae glaber is suggested. Despite a superficial resemblance, this study revealed a considerable difference of circular sphincteric muscle components between female dog and woman suggesting that functional studies in respect to urinary continence obtained in dogs cannot be attributed without qualification to humans. PMID- 11731704 TI - In vivo mechanical properties of proximal and distal aponeuroses in human tibialis anterior muscle. AB - Load-strain characteristics of distal (deep) and proximal (superficial) aponeuroses were determined in vivo for human tibialis anterior muscle (TA). Seven male subjects exerted isometric dorsiflexion torque from relaxation to voluntary maximum while elongation of both aponeuroses of TA was determined by ultrasonography. Two positions (end of the muscle belly and a proximal part) and one position (distal part) were scanned for the deep and superficial aponeuroses, respectively, and tendinous movements of the respective positions were determined. Based on the tendinous movements, elongation of each aponeurosis was determined. Both aponeuroses were elongated significantly, and there was no significant difference in strain between the deep (3.3 +/- 0.8%) and superficial (3.0 +/- 0.5%) aponeuroses. In addition, there was a significant linear relationship between strain of deep and superficial aponeuroses. It was suggested that both aponeuroses similarly act as an elastic component in pennate muscles. PMID- 11731705 TI - Evidence for functional partitioning of the rabbit digastric muscle. AB - The rabbit digastric muscle has a single belly that opens and retracts the mandible. It does not contain connective tissue partitions, and all fibers arise from the same tendon and insert into a single broad site. Historically, it was assumed that the muscle functioned as a single unit. Since we had preliminary evidence that this might not be the case, we carried out five small studies in rabbits. First, we showed that electromyographic (EMG) activity varies between recording sites within the muscle during the masticatory cycle induced by repetitive stimulation of the sensorimotor cortex. We found that EMG activity in the caudal region sometimes began before the anterior EMG during mastication when the jaw swung to the side of the muscle, but the two regions became active at the same time during other patterns. We next showed that separate branches of the mylohyoid nerve enter the anterior, intermediate and caudal regions of the digastric. However, a separate study showed that the motor endplates were distributed across a continuous sheet, consistent with a single anatomical partition. We then stimulated single nerve branches to deplete glycogen. By comparing the optical density of fibers labeled by the periodic acid-Schiff method for glycogen, we were able to show that the three branches innervate separate regions of the muscle. Finally, we applied either FluoroGold or Fast Blue dyes to the central cut ends of the branches to label the cell bodies of the three pools of motoneurons. These were found within the middle and caudal thirds of the trigeminal motor nucleus, but there appeared to be no spatial separation of the three pools or double labeling of cells. We conclude that the digastric muscle contains two and possibly three functional subregions. The fact that the motoneurons are intermingled suggests that the distribution of motor commands to the three pools is not based on their location. PMID- 11731706 TI - Anatomical partitioning of three human forearm muscles. AB - Anatomical partitioning has been found in the human biceps brachii, extensor carpi radialis longus and flexor carpi radialis muscles. The purpose of this study was to determine if the human extensor carpi ulnaris, flexor carpi ulnaris and flexor digitorum profundus are anatomically partitioned. Evidence for or against anatomical partitioning was obtained by observation of the architectural and innervation characteristics of each of the investigated muscles. Twelve samples (11 were used for extensor carpi ulnaris) of each specific muscle type were harvested from perfused human cadavers. The architectural characteristics of tendinous boundaries, muscle fiber direction, and muscle fiber angle magnitude were observed, measured and documented. Microdissection technique was used to investigate the primary nerve branching pattern throughout each muscle. A primary nerve branch to a specific muscle region indicated possible partitioning by innervation. The extensor carpi ulnaris was found to have a variable number of primary nerve branches. The extensor carpi ulnaris may have four partitions by innervation alone or three congruent partitions by innervation and muscle fiber architecture. The nerve to the flexor carpi ulnaris clearly innervates two architectural partitions within the muscle. The innervation pattern to the flexor carpi ulnaris is congruent with muscle fiber architecture characteristics indicating consistent anatomical partitioning within the flexor carpi ulnaris. Two muscle nerves innervate the flexor digitorum profundus with branches innervating the medial and lateral regions of the muscle. Up to eight architectural partitions were found in a medial-to-lateral direction. PMID- 11731707 TI - Anthropometric analysis of the nose in the Japanese. AB - To quantify the shape of cleft lip noses and to evaluate the outcome following rhinoplasty objectively, a three-dimensional measuring system for the nasal shape was developed using a contact-type measuring apparatus. In this system, the surface data for a face could be obtained accurately, and facial landmarks could be identified semiautomatically. A new method for evaluating the nasal shape, which regards the nose as a polyhedron constructed of triangles which consist of facial landmarks, has been subsequently devised. Facial plaster models of 129 Japanese volunteers, 61 men and 68 women, were made. The nasal shapes were characterized by linear and angular measurements using semiautomatically identified landmarks, and the new polyhedral system of measurement was applied. The polyhedron was constructed of 10 triangular planes each consisting of three landmarks. The normal vector of each plane was calculated, and the angles described by these normal vectors were calculated. Consequently, the nasal shapes of males and females were characterized as follows: (1) the proportion of the size of the nasal ala to that of the nasal dorsum was larger in males than in females; (2) a more outward projection of the nasal ala was observed in males than in females; (3) the so-called 'nasal tip angle' was more acute in males than in females; (4) the transitional area from the nasal dorsum to the face was steeper in females than in males; (5) the upper compartments of the nasal ala were steeper in females than in males. PMID- 11731708 TI - Negligible effects of depression on verbal and spatial performance in Alzheimer's disease. AB - We examined whether a diagnosis of depression affects verbal and visuospatial performance in Alzheimer's disease (AD). Using data from a population-based study, persons with AD and depression (AD/D), AD alone and a control group of normal older adults were compared in two tests of verbal ability (category and letter fluency) and two tests of visuospatial skill (block design and clock drawing). As expected, there were clear AD-related deficits across all cognitive tasks. More importantly, the AD and AD/D groups were indistinguishable on all task variables. The lack of effects of depression was discussed relative to the view that those symptoms of this disease which are especially detrimental to cognitive functioning (e.g. concentration difficulties, lack of interest, loss of energy) may already be present in AD as a result of the neurodegenerative process. PMID- 11731709 TI - Prevalence and incidence of major depressive disorder in Alzheimer's disease: findings from two databases. AB - BACKGROUND: Reports on the frequency of major depression in Alzheimer's disease are conflicting, some suggesting that it is frequent, others suggesting that it is uncommon. METHODS: We examined the prevalence and incidence of symptom clusters meeting criteria for major depressive disorder (MDD) in two large series of cases diagnosed prospectively by NINCDS/ADRDA criteria as probable or possible Alzheimer's disease. Standard DSM-III-R criteria were employed at one site, the CERAD modification of DSM-III-R criteria at the other. Both sites required the presence of depressed mood on direct patient examination at the time of initial evaluation in order to meet criteria for MDD. RESULTS: At the center using standard DSM-III-R criteria (n = 329), there was a 0.9% prevalence of symptom clusters meeting criteria for MDD. At the center using the CERAD modification (n = 586), MDD prevalence was 4.8%. Incidence rates at both centers were below 2% per year over 3 years. CONCLUSIONS: We concluded that DSM-III-R MDD is not common in AD patients. PMID- 11731710 TI - The Hopkins Verbal Learning Test and screening for dementia. AB - The present study investigated the sensitivity and specificity of the Hopkins Verbal Learning Test (HVLT) for demented patients (n = 82, using NINCDS criteria) and 114 healthy controls--equivalent in age, years of education and gender-ratio- from the Oxford Project to Investigate Memory and Ageing. The HVLT 'Total Recall' score had 87% sensitivity and 98% specificity for dementia using a cut-off score of 14.5. Using a 'Memory' score (the sum of the 'Total Recall' and the 'Discrimination Index') with a cut-off score of 24.5 gave a 91% sensitivity and 98% specificity for Alzheimer's disease cases when compared to controls. Unlike the MMSE, the HVLT has no ceiling effects and does not have to be adjusted for education. We conclude that the HVLT is an easy to administer, quick and well tolerated tool for the screening of dementia. PMID- 11731711 TI - The Alzheimer's Disease Assessment Scale: findings from a low-education population. AB - We constructed a Chinese version of the cognitive component of the Alzheimer's Disease Assessment Scale (ADAS-Cog). In order to accommodate illiteracy, the Chinese version used pictures instead of words for assessing recall and recognition. The Chinese ADAS-Cog was administered to 125 individuals with no dementia, 127 with questionable dementia, and 77 with Alzheimer's disease (AD). Their age range was 51-92 years and their education range was 0-20 years. The Chinese ADAS-Cog had high internal consistency (Cronbach's alpha = 0.87) and very high interrater reliability (intraclass correlation coefficient, or ICC, = 0.99) and test-retest reliability (ICC = 0.96). It had high correlations with scores on the Clinical Dementia Rating Scale (Pearson's r = 0.85), the Cognitive Abilities Screening Instrument (CASI, Pearson's r = -0.88), and CASI-estimated scores on the Mini-Mental State Examination (Pearson's r = -0.85). Performance on the Chinese ADAS-Cog was uninfluenced by age or gender, nor by education level except within the low education range of 0-6 years. Its memory items were best for early detection of dementia; its language items were best for monitoring the progression of dementia. This study found that the Chinese ADAS-Cog is a good instrument for use with Chinese AD patients. PMID- 11731712 TI - Event-related evoked potential P300 in frontotemporal dementia. AB - There are no studies on event-related cognitive potentials in frontotemporal dementia (FTD). In order to evaluate the aptitude and usefulness of the event related P300 potential in this disease, we prospectively examined 60 cases: 11 patients with FTD diagnosed according to the Lund and Manchester criteria and Neary consensus criteria, 33 patients with a probable Alzheimer's disease diagnosis following NINCDS-ADRDA criteria, and 16 normal controls. P300 latency, amplitude and reaction time were recorded using an auditory oddball paradigm. In this sample, P300 potential could be reliably performed by 10/11 FTD patients, notwithstanding their language or executive function deficiencies. The FTD group P300 mean latency was midway between the normal controls and the Alzheimer's disease group (ANOVA F(2, 74199) = 16.5; p = 0.00003). The latency range of the FTD patients were within normal values (average plus 1.96 standard deviation of the values of the control group), except for one case with a latency of 448 ms. Post hoc Newman-Keuls analysis showed that the P300 latencies of the control and FTD groups did not differ significantly (p = 0.15) and that the Alzheimer's disease group had a delayed P300 latency that differed significantly from that of the FTD (p = 0.002) and control group (p = 0.0002). However, there was overlapping in P300 latency values of the three groups. Despite these differences in latencies, the reaction time was significantly increased in the FTD and the Alzheimer's disease groups. These findings indicate that the P300 potential is less affected in patients with FTD than those with Alzheimer's disease. This fact could aid in FTD diagnosis, differential diagnosis with Alzheimer's disease and possibly its clinical management. PMID- 11731713 TI - Economic evaluation of donepezil treatment for Alzheimer's disease in Japan. AB - To demonstrate the economic impact of donepezil treatment for patients with mild to moderate Alzheimer's disease (AD) in Japanese clinical settings, we conducted a cost-utility analysis using the Markov model. This analysis was made from a payer's perspective, and the cost was estimated based on the fee schedule of the Japanese national health insurance and the long-term care insurance. The results of the clinical trial of donepezil on AD in Japan were used to estimate the drug efficacy. Under the base-case analysis with a 2-year time horizon, donepezil treatment for patients with mild to moderate AD was considered to be an economically dominant strategy compared to conventional therapy. PMID- 11731714 TI - The effect of the living situation on the severity of dementia at diagnosis. AB - INTRODUCTION: Previous studies have shown that social determinants not directly involved in the disease process may be implicated in the timing of dementia diagnosis. This study explores the relationship between a patient's living situation and the severity of dementia at diagnosis. METHODS: Data were collected from the baseline interviews of 1,325 patients with cognitive decline enrolled in the Consortium for the Investigation of Vascular Impairment of Cognition study. Data collected included: age, sex, living situation and scores on the Mini-Mental State Examination (MMSE), Global Deterioration Scale (GDS), the Functional Rating Scale (FRS), the Disability Assessment for Dementia (DAD) scale and the Cumulative Illness Rating Scale (CIRS). Living situation was grouped as: (1) lives alone, (2) lives with spouse, (3) lives with child, relative or other and (4) lives in a nursing home. A general linear model univariate analysis was used to compare patients by their respective living situations for differences in mean scores on each of the 4 measures of dementia severity. RESULTS: Statistical analysis of both unadjusted data and data adjusted for age, sex and CIRS scores showed significant differences among the groups. Those who lived alone were diagnosed at an earlier stage (mean scores: MMSE 21.4, GDS 3.6, FRS 20.0, DAD 29.8) followed by those who lived with a spouse (mean scores: MMSE 20.5, GDS 3.7, FRS 20.4, DAD 28.0), those who lived with a child or other (mean scores: MMSE 19.3, GDS 3.9, FRS 22.5, DAD 24.9) and finally those who lived in a nursing home (mean scores: MMSE 15.2, GDS 4.8, FRS 27.5, DAD 16.9). CONCLUSION: Living situation is related to the severity of dementia at diagnosis. Primary care providers should have a low threshold for case-finding in older adults who live with family or friends. PMID- 11731715 TI - Age- and dementia-associated impairments in divided attention: psychological constructs, animal models, and underlying neuronal mechanisms. AB - Following a brief description of the psychological construct 'divided attention', impairments in divided attention and related executive functions are discussed as the major variable in the development and manifestation of age- and dementia associated cognitive impairments. Neuropsychological and functional imaging studies in humans have indicated that dorsolateral and ventrolateral prefrontal, cingulate, parietal and premotor cortical areas are involved in the mediation of dual task performance. Furthermore, reduced activity in these areas has been suggested to mediate age- and dementia-associated impairments in divided attention. Experimental studies in animals have provided strong support for the hypothesis that cholinergic projections terminating in all cortical areas and layers crucially mediate the performance in tasks that tax processing capacity and/or the allocation of processing resources to competing demands. This specification of the 'cholinergic hypothesis' is evaluated in light of recent critical accounts of the role of this system in the development of age- and dementia- related cognitive disorders. The converging animal experimental and human neuropathological, as well as neuropsychological, evidence indicates that decreases in the integrity of cortical cholinergic inputs represent a necessary, possibly even sufficient, neuronal process mediating the impairments in divided attention and the resulting, broad decline in cognitive functions. PMID- 11731716 TI - Progress in the search for ideal drugs. AB - The search for ideal drugs to improve patient care requires applications of modern scientific methods to both discovery and development. Using these modern methods, the pharmaceutical industry with strong academic and government collaboration has introduced in the last 25 years many new drugs that approach the ideal. After reviewing Bjornsson's classification of drug action and the notion of contributory causality, this commentary defines an ideal drug from the perspectives of pharmacodynamics, pharmacokinetics, and therapeutics. Examples of new drugs for hypertension, heart disease, stroke, osteoporosis, asthma, ulcer, and migraine headaches are described. Finally, the profound implications of progress in developing ideal drugs not only for the patient but also for the academic, educational, and regulatory establishments are briefly reviewed. PMID- 11731717 TI - Reduction of submissive behavior in rats: a test for antidepressant drug activity. AB - Randomly paired rats were food deprived overnight and placed in an apparatus compelling them to compete for a food reward. About half of these pairs developed a dominant-submissive relationship measured as a significant difference in time spent on the feeder by each rat. This relationship developed over a 2-week period and remained stable for at least the next 5 weeks. Treatment of the submissive subjects, for at least 2 weeks, with imipramine, desipramine, or fluoxetine (10 mg/kg) significantly reduced submissive behavior. The effect faded after cessation of treatment with desipramine. Fluoxetine was further tested at 2.5- and 5-mg/kg doses and showed a dose-dependent reduction of submissive behavior. Treatment of submissive rats with the anxiolytic diazepam (1 mg/kg) was ineffective. The prevalence of dominant-submissive relationships and the effect of desipramine and imipramine on submissive behavior were gender independent. The predictive, face, and construct validity of the behavioral test is discussed. PMID- 11731718 TI - Effects of silymarin MZ-80 on hepatic oxidative stress in rats with biliary obstruction. AB - This study was designed to evaluate the effects of three pharmaceutical forms of silymarin (silymarin MZ-80, silybinin-beta-cyclodextrin, and silybinin) on the liver oxidative status in vitro and after oral administration to rats with extrahepatic biliary obstruction (EBO) and sham-operated animals. We evaluated thiobarbituric acid-reactive substances (TBARS), glutathione (GSH + GSSG) and their related enzyme activities (GSH peroxidase, GSSG reductase and GSH transferase). All three compounds inhibited the in vitro production of TBARS (IC(50) 56-533 micromol/l). These compounds, mainly silymarin MZ-80, also increased GSH peroxidase and GSH transferase activities. In EBO rats we found increases in TBARS production which was inhibited by 50-70% after treatment. Glutathione was reduced by 55% and elevated by silymarin MZ-80. GSH transferase increased in the group given silymarin MZ-80. We conclude that all three derivatives of silymarin show a clear ability to reduce lipid peroxidation in the liver. Silymarin MZ-80 was the only compound that enhanced the glutathione antioxidant system. PMID- 11731719 TI - Effect of rebamipide on cell death induced by combined treatment of mild heat shock and quercetin in RGM-1 cells: a role for HSP70 induction. AB - The mechanism by which rebamipide inhibits cell death induced by combined treatment of mild heat shock and quercetin in RGM-1 cells was examined. Cells were incubated at 42 degrees C for 1 h with or without quercetin. Expression of HSP70 was detected by immunoblot analysis and cell viability was determined by the MTT method. Upon treatment of cells with heat shock in the presence of quercetin, cell viability was significantly decreased 24 h after heat shock, with strong inhibition of HSP70 expression. Rebamipide suppressed both cell death and a reduction in HSP70 induced by the combined treatment of heat shock and quercetin. The combination of heat shock and quercetin resulted in DNA fragmentation, but rebamipide managed to inhibit such fragmentation. The protective effect of rebamipide was eliminated by cycloheximide or actinomycin D. Based upon these findings, it was concluded that rebamipide inhibited cell death resulting from combined treatment of mild heat shock and quercetin. The protective action of rebamipide appears to be at least partially due to maintenance of the HSP70 level. PMID- 11731720 TI - Effects of HNS-32, a novel antiarrhythmic agent, on guinea-pig myocardium. AB - The electrophysiological and mechanical effects of HNS-32, a novel azulene-1 carboxamidine derivative with antiarrhythmic activity, were studied in isolated guinea-pig myocardial preparations. HNS-32 (10(-6)-10(-4) mol/l) concentration dependently decreased the maximum rate of rise (V(max)) of action potential in isolated papillary muscle; the potency was the same or slightly higher than that of disopyramide. At 10(-4) mol/l, HNS-32 also shortened the action potential duration (APD) and depolarized the resting membrane potential; these effects were similar to those of 10(-5) mol/l verapamil. HNS-32 (10(-7)-10(-4) mol/l), as well as verapamil (10(-8)-10(-5) mol/l) and disopyramide (10(-6)-10(-3) mol/l), had concentration-dependent negative chronotropic and negative inotropic effects on isolated right atrial and right ventricular papillary muscle preparations, respectively. The concentration-response relationship for the positive chronotropic effect of isoproterenol was not affected by HNS-32 (10(-5) mol/l). In isolated ventricular myocytes, HNS-32 (10(-6)-10(-4) mol/l) concentration dependently inhibited the peak amplitude of the L-type Ca(2+) current. These results suggest that NHS-32 has V(max) reducing activity on myocardial tissue, which may be responsible for antiarrhythmic effect. The drug may also have additional effect on the Ca(2+) channel at higher concentrations. PMID- 11731721 TI - Calcium-antagonistic activity of sumatriptan in the rat anococcygeus muscle. AB - The relaxant effect of sumatriptan, a 5-HT(1B/1D) receptor agonist, on the rat anococcygeus muscle was investigated. Sumatriptan induced concentration-dependent relaxations of the phenylephrine-precontracted rat anococcygeus muscle. N(G) nitro-L-arginine, methylene blue, glibenclamide, tetrodotoxin, indomethacin, GR 113808, GR 55562, methysergide, ketanserin, ICS 205930, clozapine, methiothepin, metergoline, mesulergine, and ritanserin did not inhibit the relaxations induced by sumatriptan. Sumatriptan converted calcium-induced contractions into relaxations in the preparations depolarized by a calcium-free, high-potassium solution. In membrane preparations obtained from the rat anococcygeus muscle, the basal rate of cAMP production by adenylate cyclase was 4.77 +/- 0.02 pmol/mg protein/min (n = 15). The enzyme activity was increased to 104 +/- 51.7 pmol/mg protein/min by forskolin (10(-4) mol/l), but did not change in the presence of sumatriptan. The mRNA expression of 5-HT(1B), 5-HT(1D), and 5-HT(7) receptors was not observed in the rat anococcygeus muscle. The results indicate that sumatriptan causes relaxation of the precontracted rat anococcygeus muscle by a calcium-antagonistic activity. PMID- 11731723 TI - The broad clinical activity of ECT should not be ignored. PMID- 11731722 TI - Acetylcholine attenuates cardiomyocyte oxidant stress during simulated ischemia and reoxygenation. AB - We wanted to determine whether oxygen radicals open the mitochondrial ATP dependent potassium channels (K(ATP)) during an ischemic period to reduce cell death and oxidant stress. Chick embryonic cardiomyocytes were used. Cell viability was quantified with propidium iodide (5 microM), and free radicals was measured using 2',7'-dichlorofluorescein diacetate. Preconditioning was produced by 10 min of simulated ischemia followed by 10 min of reoxygenation. Acetylcholine (1 mM), infused for 10 min instead of preconditioning, reduced cell death similarly (24 +/- 5%, n = 7 and 18 +/- 2%, n = 7, respectively, vs. controls, 49 +/- 6%, n = 8). In control series, 60 min of simulated ischemia and 3 h of reoxygenation generated free radicals more than 300% above the baseline (ischemia: 3.63 +/- 0.58, reoxygenation: 3.66 +/- 0.47, n = 8). Preconditioning and acetylcholine markedly attenuated the oxidant stress during simulated ischemia (1.18 +/- 0.41, n = 6 and 1.34 +/- 0.60, n = 7 vs. controls 3.63 +/- 0.58, n = 8) and re-oxygenation (1.23 +/- 0.36, n = 6 and 1.50 +/- 0.59, n = 7 vs. controls 3.66 +/- 0.47, n = 8). The protection of acetylcholine was abolished with pretreatment with the antioxidant thiol reductant 2-mercaptopropionyl glycine and posttreatment with 5-hydroxydecanoate, a selective mitochondrial K(ATP) channel antagonist (37 +/- 7%, n = 7). These results demonstrate that oxygen radicals open mitochondrial K(ATP) channels, which mediates the acetylcholine-induced preconditioning effect, and that stimulation of this signaling pathway attenuates oxidant stress. PMID- 11731724 TI - ECT in mental retardation: a review. AB - The use of electroconvulsive therapy (ECT) in patients with mental retardation and a psychiatric disorder seems to be infrequent, and controlled studies are not available. We reviewed the literature on the use of ECT in mental retardation, using Medline, Embase, and the Cochrane Library. References were checked and ECT experts consulted. The reported case studies were examined on patients' characteristics and illness, treatment, and outcome variables. Forty-four patients, mostly with a diagnosis of psychotic depression, were found. In 84% (n = 37) of them, ECT was effective and without important side effects. However, relapse occurred frequently, in 48% (n = 21) of the patients. In 16% (n = 7), side effects and/or no improvement were noted. The reported case studies suggest that ECT may be of value in treating severe psychiatric disorders in mentally retarded patients, with similar indications as in general psychiatry. However, the lack of strong scientific evidence, besides complicated psychiatric assessment as well as ethical and legal issues, probably cause an unnecessarily limited use of ECT in these patients. Further controlled trails are needed to firmly establish the efficacy and safety of ECT in mental retardation. PMID- 11731725 TI - ECT remission rates in psychotic versus nonpsychotic depressed patients: a report from CORE. AB - OBJECTIVE: To compare the relative efficacy of electroconvulsive therapy (ECT) in psychotic and nonpsychotic patients with unipolar major depression. METHODS: The outcome of an acute ECT course in 253 patients with nonpsychotic (n = 176) and psychotic (n = 77) unipolar major depression was assessed in the first phase of an ongoing National Institute of Mental Health-supported four-hospital collaborative study of continuation treatments after successful ECT courses. ECT was administered with bilateral electrode placement at 50% above the titrated seizure threshold. The remission criteria were rigorous: a score 7 days (OR 4.29, 95% CI 1.82-10.1), SAPS II score >30 (OR 3.34, 95% CI 1.0-11.18), total parenteral nutrition (OR 2.69, 95% CI 1.19-6.07) and tracheostomy (OR 1.88, 95% CI 0.84-4.20). CONCLUSIONS: Nosocomial infections are relatively frequent in Triveneto area ICUs. The predominance of pneumonia and bloodstream infection, and the high frequency of antibiotic-resistant pathogens indicate that resources had to be assigned towards the implementation of control programs of those infections, monitoring of antibiotic resistance and prescription, and antibiotic therapy guidelines. PMID- 11731757 TI - The treatment of Reflex Sympathetic Dystrophy in a 9 year-old boy with long standing symptoms. AB - Reflex sympathetic dystrophy is an uncommonly reported entity in children and it continues to be underdiagnosed. Compared with adult, childhood reflex sympathetic dystrophy is of unknown etiology and has a better prognosis. The most common therapy in children is progressive mobilization supported by antiphlogistic, analgesic drugs, psychological and physical therapy. We report an interesting case of reflex sympathetic dystrophy of the left knee joint of a nine years old child with symptoms insisting more than four years and recalcitrant to the above treatments. The use of intravenous regional anaesthesia with lidocaine 0.5% and methylprednisolone was successful. No other reports seem to exist on the use of lidocaine 0.5% and methylprednisolone for the therapy of reflex sympathetic dystrophy in children. The treatment is simple, safe and well tolerated by children. Psychological factors should not be underestimated. Early diagnosis and aggressive therapy are important factors for the full recovery of the patients. PMID- 11731758 TI - [The European certificate of anesthesia and intensive care]. PMID- 11731760 TI - Dreams and reality. PMID- 11731759 TI - Beaten out of submission. PMID- 11731761 TI - French loan arranger seeks biotech posse. PMID- 11731762 TI - Illegal Bt cotton in India haunts regulators. PMID- 11731763 TI - US anti-bioterror efforts swiftly expanding. PMID- 11731764 TI - New Zealand's compromise calms GMO lobby. PMID- 11731765 TI - Sleeker Incyte flirts with drug discovery. PMID- 11731767 TI - Realism in drug discovery-could Cassandra be right? PMID- 11731770 TI - Follow my leader. PMID- 11731771 TI - Plotting a course for GM forestry. PMID- 11731772 TI - Grafting for transgene containment. PMID- 11731773 TI - Regulation and exploitation of genetically modified crops. PMID- 11731774 TI - Can bioremediation bounce back? PMID- 11731775 TI - Stem cells with brainpower. PMID- 11731776 TI - Nuclear security breached. PMID- 11731777 TI - Pink water, green plants, and pink elephants. PMID- 11731778 TI - Multiplex metallica. PMID- 11731779 TI - Beyond comparison--antibiotics from genome data? PMID- 11731781 TI - In vitro differentiation of transplantable neural precursors from human embryonic stem cells. AB - The remarkable developmental potential and replicative capacity of human embryonic stem (ES) cells promise an almost unlimited supply of specific cell types for transplantation therapies. Here we describe the in vitro differentiation, enrichment, and transplantation of neural precursor cells from human ES cells. Upon aggregation to embryoid bodies, differentiating ES cells formed large numbers of neural tube-like structures in the presence of fibroblast growth factor 2 (FGF-2). Neural precursors within these formations were isolated by selective enzymatic digestion and further purified on the basis of differential adhesion. Following withdrawal of FGF-2, they differentiated into neurons, astrocytes, and oligodendrocytes. After transplantation into the neonatal mouse brain, human ES cell-derived neural precursors were incorporated into a variety of brain regions, where they differentiated into both neurons and astrocytes. No teratoma formation was observed in the transplant recipients. These results depict human ES cells as a source of transplantable neural precursors for possible nervous system repair. PMID- 11731782 TI - Neural progenitors from human embryonic stem cells. AB - The derivation of neural progenitor cells from human embryonic stem (ES) cells is of value both in the study of early human neurogenesis and in the creation of an unlimited source of donor cells for neural transplantation therapy. Here we report the generation of enriched and expandable preparations of proliferating neural progenitors from human ES cells. The neural progenitors could differentiate in vitro into the three neural lineages--astrocytes, oligodendrocytes, and mature neurons. When human neural progenitors were transplanted into the ventricles of newborn mouse brains, they incorporated in large numbers into the host brain parenchyma, demonstrated widespread distribution, and differentiated into progeny of the three neural lineages. The transplanted cells migrated along established brain migratory tracks in the host brain and differentiated in a region-specific manner, indicating that they could respond to local cues and participate in the processes of host brain development. Our observations set the stage for future developments that may allow the use of human ES cells for the treatment of neurological disorders. PMID- 11731783 TI - Magnetodendrimers allow endosomal magnetic labeling and in vivo tracking of stem cells. AB - Magnetic resonance (MR) tracking of magnetically labeled stem and progenitor cells is an emerging technology, leading to an urgent need for magnetic probes that can make cells highly magnetic during their normal expansion in culture. We have developed magnetodendrimers as a versatile class of magnetic tags that can efficiently label mammalian cells, including human neural stem cells (NSCs) and mesenchymal stem cells (MSCs), through a nonspecific membrane adsorption process with subsequent intracellular (non-nuclear) localization in endosomes. The superparamagnetic iron oxide nanocomposites have been optimized to exhibit superior magnetic properties and to induce sufficient MR cell contrast at incubated doses as low as 1 microg iron/ml culture medium. When containing between 9 and 14 pg iron/cell, labeled cells exhibit an ex vivo nuclear magnetic resonance (NMR) relaxation rate (1/T2) as high as 24-39 s-1/mM iron. Labeled cells are unaffected in their viability and proliferating capacity, and labeled human NSCs differentiate normally into neurons. Furthermore, we show here that NSC-derived (and LacZ-transfected), magnetically labeled oligodendroglial progenitors can be readily detected in vivo at least as long as six weeks after transplantation, with an excellent correlation between the obtained MR contrast and staining for beta-galactosidase expression. The availability of magnetodendrimers opens up the possibility of MR tracking of a wide variety of (stem) cell transplants. PMID- 11731784 TI - In vivo near-infrared fluorescence imaging of osteoblastic activity. AB - In vertebrates, the development and integrity of the skeleton requires hydroxyapatite (HA) deposition by osteoblasts. HA deposition is also a marker of, or a participant in, processes as diverse as cancer and atherosclerosis. At present, sites of osteoblastic activity can only be imaged in vivo using gamma emitting radioisotopes. The scan times required are long, and the resultant radioscintigraphic images suffer from relatively low resolution. We have synthesized a near-infrared (NIR) fluorescent bisphosphonate derivative that exhibits rapid and specific binding to HA in vitro and in vivo. We demonstrate NIR light-based detection of osteoblastic activity in the living animal, and discuss how this technology can be used to study skeletal development, osteoblastic metastasis, coronary atherosclerosis, and other human diseases. PMID- 11731785 TI - Selective enhancement of gene transfer by steroid-mediated gene delivery. AB - The incorporation of transgenes into the host cells' nuclei is problematic using conventional nonviral gene delivery technologies. Here we describe a strategy called steroid-mediated gene delivery (SMGD), which uses steroid receptors as shuttles to facilitate the uptake of transfected DNA into the nucleus. We use glucocorticoid receptors (GRs) as a model system with which to test the principle of SMGD. To this end, we synthesized and tested several bifunctional steroid derivatives, finally focusing on a compound named DR9NP, consisting of a dexamethasone backbone linked to a psoralen moiety using a nine-atom chemical spacer. DR9NP binds to the GR in either its free or DNA-crosslinked form, inducing the translocation of the GR to the nucleus. The expression of transfected DR9NP-decorated reporter plasmids is enhanced in dividing cells: expression of steroid-decorated reporter plasmids depends on the presence of the GR, is independent of the transactivation potential of the GR, and correlates with enhanced nuclear accumulation of the transgene in GR-positive cells. The SMGD effect is also observed in cells naturally expressing GRs and is significantly increased in nondividing cell cultures. We propose that SMGD could be used as a platform for selective targeting of transgenes in nonviral somatic gene transfer. PMID- 11731786 TI - Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria. AB - Mobile group II introns can be retargeted to insert into virtually any desired DNA target. Here we show that retargeted group II introns can be used for highly specific chromosomal gene disruption in Escherichia coli and other bacteria at frequencies of 0.1-22%. Furthermore, the introns can be used to introduce targeted chromosomal breaks, which can be repaired by transformation with a homologous DNA fragment, enabling the introduction of point mutations. Because of their wide host range, mobile group II introns should be useful for genetic engineering and functional genomics in a wide variety of bacteria. PMID- 11731787 TI - Phytodetoxification of TNT by transgenic plants expressing a bacterial nitroreductase. AB - There is major international concern over the wide-scale contamination of soil and associated ground water by persistent explosives residues. 2,4,6 Trinitrotoluene (TNT) is one of the most recalcitrant and toxic of all the military explosives. The lack of affordable and effective cleanup technologies for explosives contamination requires the development of better processes. Significant effort has recently been directed toward the use of plants to extract and detoxify TNT. To explore the possibility of overcoming the high phytotoxic effects of TNT, we expressed bacterial nitroreductase in tobacco plants. Nitroreductase catalyzes the reduction of TNT to hydroxyaminodinitrotoluene (HADNT), which is subsequently reduced to aminodinitrotoluene derivatives (ADNTs). Transgenic plants expressing nitroreductase show a striking increase in ability to tolerate, take up, and detoxify TNT. Our work suggests that expression of nitroreductase (NR) in plants suitable for phytoremediation could facilitate the effective cleanup of sites contaminated with high levels of explosives. PMID- 11731788 TI - A peptide carrier for the delivery of biologically active proteins into mammalian cells. AB - The development of peptide drugs and therapeutic proteins is limited by the poor permeability and the selectivity of the cell membrane. There is a growing effort to circumvent these problems by designing strategies to deliver full-length proteins into a large number of cells. A series of small protein domains, termed protein transduction domains (PTDs), have been shown to cross biological membranes efficiently and independently of transporters or specific receptors, and to promote the delivery of peptides and proteins into cells. TAT protein from human immunodeficiency virus (HIV-1) is able to deliver biologically active proteins in vivo and has been shown to be of considerable interest for protein therapeutics. Similarly, the third alpha-helix of Antennapedia homeodomain, and VP22 protein from herpes simplex virus promote the delivery of covalently linked peptides or proteins into cells. However, these PTD vectors display a certain number of limitations in that they all require crosslinking to the target peptide or protein. Moreover, protein transduction using PTD-TAT fusion protein systems may require denaturation of the protein before delivery to increase the accessibility of the TAT-PTD domain. This requirement introduces an additional delay between the time of delivery and intracellular activation of the protein. In this report, we propose a new strategy for protein delivery based on a short amphipathic peptide carrier, Pep-1. This peptide carrier is able to efficiently deliver a variety of peptides and proteins into several cell lines in a fully biologically active form, without the need for prior chemical covalent coupling or denaturation steps. In addition, this peptide carrier presents several advantages for protein therapy, including stability in physiological buffer, lack of toxicity, and lack of sensitivity to serum. Pep-1 technology should be extremely useful for targeting specific protein-protein interactions in living cells and for screening novel therapeutic proteins. PMID- 11731789 TI - Production of calves from G1 fibroblasts. AB - Since the landmark study of Wilmut et al. describing the birth of a cloned lamb derived from a somatic cell nucleus, there has been debate about the donor nucleus cell cycle stage required for somatic cell nuclear transfer (NT). Wilmut et al. suggested that induction of quiescence by serum starvation was critical in allowing donor somatic cells to support development of cloned embryos. In a subsequent report, Cibelli et al. proposed that G0 was unnecessary and that calves could be produced from actively dividing fibroblasts. Neither study conclusively documented the importance of donor cell cycle stage for development to term. Other laboratories have had success with NT in several species, and most have used a serum starvation treatment. Here we evaluate methods for producing G0 and G1 cell populations and compare development following NT. High confluence was more effective than serum starvation for arresting cells in G0. Pure G1 cell populations could be obtained using a "shake-off" procedure. No differences in in vitro development were observed between cells derived from the high-confluence treatment and from the "shake-off" treatment. However, when embryos from each treatment were transferred to 50 recipients, five calves were obtained from embryos derived from "shake-off" cells, whereas no embryos from confluent cells survived beyond 180 days of gestation. These results indicate that donor cell cycle stage is important for NT, particularly during late fetal development, and that actively dividing G1 cells support higher development rates than cells in G0. PMID- 11731790 TI - An intellectual property clearinghouse for agricultural biotechnology. PMID- 11731794 TI - Training needs in regulatory science for the biopharmaceutical industry. PMID- 11731795 TI - MLL translocations specify a distinct gene expression profile that distinguishes a unique leukemia. AB - Acute lymphoblastic leukemias carrying a chromosomal translocation involving the mixed-lineage leukemia gene (MLL, ALL1, HRX) have a particularly poor prognosis. Here we show that they have a characteristic, highly distinct gene expression profile that is consistent with an early hematopoietic progenitor expressing select multilineage markers and individual HOX genes. Clustering algorithms reveal that lymphoblastic leukemias with MLL translocations can clearly be separated from conventional acute lymphoblastic and acute myelogenous leukemias. We propose that they constitute a distinct disease, denoted here as MLL, and show that the differences in gene expression are robust enough to classify leukemias correctly as MLL, acute lymphoblastic leukemia or acute myelogenous leukemia. Establishing that MLL is a unique entity is critical, as it mandates the examination of selectively expressed genes for urgently needed molecular targets. PMID- 11731796 TI - Loss of the SKI proto-oncogene in individuals affected with 1p36 deletion syndrome is predicted by strain-dependent defects in Ski-/- mice. AB - Experiments involving overexpression of Ski have suggested that this gene is involved in neural tube development and muscle differentiation. In agreement with these findings, Ski-/- mice display a cranial neural tube defect that results in exencephaly and a marked reduction in skeletal muscle mass. Here we show that the penetrance and expressivity of the phenotype changes when the null mutation is backcrossed into the C57BL6/J background, with the principal change involving a switch from a neural tube defect to midline facial clefting. Other defects, including depressed nasal bridge, eye abnormalities, skeletal muscle defects and digit abnormalities, show increased penetrance in the C57BL6/J background. These phenotypes are interesting because they resemble some of the features observed in individuals diagnosed with 1p36 deletion syndrome, a disorder caused by monosomy of the short arm of human chromosome 1p (refs. 6-9). These similarities prompted us to re-examine the chromosomal location of human SKI and to determine whether SKI is included in the deletions of 1p36. We found that human SKI is located at distal 1p36.3 and is deleted in all of the individuals tested so far who have this syndrome. Thus, SKI may contribute to some of the phenotypes common in 1p36 deletion syndrome, and particularly to facial clefting. PMID- 11731797 TI - Merlin--rapid analysis of dense genetic maps using sparse gene flow trees. AB - Efforts to find disease genes using high-density single-nucleotide polymorphism (SNP) maps will produce data sets that exceed the limitations of current computational tools. Here we describe a new, efficient method for the analysis of dense genetic maps in pedigree data that provides extremely fast solutions to common problems such as allele-sharing analyses and haplotyping. We show that sparse binary trees represent patterns of gene flow in general pedigrees in a parsimonious manner, and derive a family of related algorithms for pedigree traversal. With these trees, exact likelihood calculations can be carried out efficiently for single markers or for multiple linked markers. Using an approximate multipoint calculation that ignores the unlikely possibility of a large number of recombinants further improves speed and provides accurate solutions in dense maps with thousands of markers. Our multipoint engine for rapid likelihood inference (Merlin) is a computer program that uses sparse inheritance trees for pedigree analysis; it performs rapid haplotyping, genotype error detection and affected pair linkage analyses and can handle more markers than other pedigree analysis packages. PMID- 11731798 TI - Multiple defects in antigen presentation and T cell development by mice expressing cytoplasmic tail-truncated CD1d. AB - For members of the CD1 family of beta(2)-microglobulin-associated lipid presenting molecules, tyrosine-based motifs in the cytoplasmic tail and invariant chain (Ii) govern glycoprotein trafficking through endosomal compartments. Little is known about the intracellular pathways of CD1 trafficking and antigen presentation. However, in vitro studies with cells transfected with mutant CD1 that had a truncated cytoplasmic tail have suggested a role for these tyrosine motifs in some, but not all, antigenic systems. By introducing a deletion of the tyrosine motif into the germ line, and through homologous recombination in embryonic stem cells, we now describe knock-in mice with the CD1d cytoplasmic tail deleted. Despite adequate surface CD1d expression and the presence of Ii, these mutant mice showed multiple and selective abnormalities in intracellular trafficking, antigen presentation and T cell development, demonstrating the critical functions of the CD1d cytoplasmic tail motif in vivo. PMID- 11731799 TI - Costimulation and endogenous MHC ligands contribute to T cell recognition. AB - To initiate an immune response, key receptor-ligand pairs must cluster in "immune synapses" at the T cell-antigen-presenting cell (APC) interface. We visualized the accumulation of a major histocompatibility complex (MHC) class II molecule, I E(k), at a T cell-B cell interface and found it was dependent on both antigen recognition and costimulation. This suggests that costimulation-driven active transport of T cell surface molecules helps to drive immunological synapse formation. Although only agonist peptide-MHC class II (agonist pMHC class II) complexes can initiate T cell activation, endogenous pMHC class II complexes also appeared to accumulate. To test this directly, we labeled a "null" pMHC class II complex and found that, although it lacked major TCR contact residues, it could be driven into the synapse in a TCR-dependent manner. Thus, low-affinity ligands can contribute to synapse formation and T cell signaling. PMID- 11731800 TI - T cells down-modulate peptide-MHC complexes on APCs in vivo. AB - T cells compete in the response to antigen in vivo and this competition may drive the affinity maturation of a secondary T cell response. Here we show that high affinity T cells out-competed lower affinity T cells during a response to antigenic challenge in vivo. Although competition between T cells specific for different peptide-major histocompatibility complexes (MHC) occurred, it was less efficient than competition between T cells of the same peptide-MHC specificity. In addition, high-affinity T cells efficiently induced antigen loss from the surface of antigen-presenting cells. Thus T cells that responded to the same peptide-MHC competed with each other by lowering the amount of ligand with which the cells could react. As a result, the activation of high-affinity cells was favored. This provides a mechanism for the affinity maturation of a secondary T cell response. PMID- 11731801 TI - Pattern-motion responses in human visual cortex. AB - Physiological models of visual motion processing posit that 'pattern-motion cells' represent the direction of moving objects independent of their particular spatial pattern. We performed fMRI experiments to identify neuronal activity in the human brain selective for pattern motion. A protocol using adaptation to moving 'plaid' stimuli allowed us to separate pattern-motion responses from other types of motion-related activity within the same brain structures, and revealed strong pattern-motion selectivity in human visual area MT+. Reducing the perceptual coherence of the plaids yielded a corresponding decrease in pattern motion responsivity, providing evidence that percepts of coherent motion are closely linked to the activity of pattern-motion cells in human MT+. PMID- 11731802 TI - Altered responsiveness to cocaine in rats exposed to methylphenidate during development. AB - Evidence in laboratory animals indicates that exposure to stimulants produces sensitization to their rewarding effects, a process that in humans would be expected to increase the risk of substance abuse. However, therapeutic administration of stimulants such as methylphenidate (MPH) in children with attention deficit hyperactivity disorder reportedly reduces the risk of substance abuse. Here we show in rats that exposure to MPH during pre-adolescence causes behavioral and neurobiological adaptations that endure into adulthood, and that are consistent with increased sensitivity to the aversive effects of cocaine. PMID- 11731803 TI - The antimalarial and cytotoxic drug cryptolepine intercalates into DNA at cytosine-cytosine sites. AB - Cryptolepine, a naturally occurring indoloquinoline alkaloid used as an antimalarial drug in Central and Western Africa, has been found to bind to DNA in a formerly unknown intercalation mode. Evidence from competition dialysis assays demonstrates that cryptolepine is able to bind CG-rich sequences containing nonalternating CC sites. Here we show that cryptolepine interacts with the CC sites of the DNA fragment d(CCTAGG)(2) in a base-stacking intercalation mode. This is the first DNA intercalator complex, from approximately 90 solved by X-ray crystallography, to bind a nonalternating (pyrimidine-pyrimidine) DNA sequence. The asymmetry of the drug induces a perfect stacking with the asymmetric site, allowing for the stability of the complex in the absence of hydrogen bonding interactions. The crystal structure of this antimalarial drug-DNA complex provides evidence for the first nonalternating intercalation and, as such, provides a basis for the design of new anticancer or antimalarial drugs. PMID- 11731805 TI - Domain alternation switches B(12)-dependent methionine synthase to the activation conformation. AB - B(12)-dependent methionine synthase (MetH) from Escherichia coli is a large modular protein that uses bound cobalamin as an intermediate methyl carrier. Major domain rearrangements have been postulated to explain how cobalamin reacts with three different substrates: homocysteine, methyltetrahydrofolate and S adenosylmethionine (AdoMet). Here we describe the 3.0 A structure of a 65 kDa C terminal fragment of MetH that spans the cobalamin- and AdoMet-binding domains, arranged in a conformation suitable for the methyl transfer from AdoMet to cobalamin that occurs during activation. In the conversion to the activation conformation, a helical domain that capped the cofactor moves 26 A and rotates by 63 degrees, allowing formation of a new interface between cobalamin and the AdoMet-binding (activation) domain. Interactions with the MetH activation domain drive the cobalamin away from its binding domain in a way that requires dissociation of the axial cobalt ligand and, thereby, provide a mechanism for control of the distribution of enzyme conformations. PMID- 11731806 TI - Our human genome--how can it serve us well? PMID- 11731804 TI - The crystal structure of spermidine synthase with a multisubstrate adduct inhibitor. AB - Polyamines are essential in all branches of life. Spermidine synthase (putrescine aminopropyltransferase, PAPT) catalyzes the biosynthesis of spermidine, a ubiquitous polyamine. The crystal structure of the PAPT from Thermotoga maritima (TmPAPT) has been solved to 1.5 A resolution in the presence and absence of AdoDATO (S-adenosyl-1,8-diamino-3-thiooctane), a compound containing both substrate and product moieties. This, the first structure of an aminopropyltransferase, reveals deep cavities for binding substrate and cofactor, and a loop that envelops the active site. The AdoDATO binding site is lined with residues conserved in PAPT enzymes from bacteria to humans, suggesting a universal catalytic mechanism. Other conserved residues act sterically to provide a structural basis for polyamine specificity. The enzyme is tetrameric; each monomer consists of a C-terminal domain with a Rossmann-like fold and an N terminal beta-stranded domain. The tetramer is assembled using a novel barrel type oligomerization motif. PMID- 11731807 TI - A national register for surveillance of inherited disorders: beta thalassaemia in the United Kingdom. AB - OBJECTIVE: To demonstrate the value of a national register for surveillance of services for an inherited disorder. METHODS: Data from the United Kingdom Thalassaemia Register and the United Kingdom Register of Prenatal Diagnosis for Haemoglobin Disorders were combined in a database; these registers include all fetuses known to have been diagnosed with beta thalassaemia major, beta thalassaemia intermedia, or haemoglobin E/beta thalassaemia in the United Kingdom. Data were extracted to show outcomes (selective abortion or live birth) of all fetuses and the status of those born with a disorder (alive, dead, successful bone marrow transplant, or lost to follow-up) by parents' region of residence and ethnicity. FINDINGS: At the end of 1999 the register included 1074 patients, 807 of whom were alive and residing in the United Kingdom. A successful bone marrow transplant has been performed for 117 out of 581 (20%) patients born since 1975. Residents of Pakistani origin are now the main group at risk in the United Kingdom, replacing residents of Cypriot origin. This has led to a marked shift in the need for services from the south-east of England to the Midlands and the north of England. Despite the acceptability of prenatal diagnosis, the proportion of affected births remains 50% higher than would be expected, reflecting a widespread failure to deliver timely screening and counselling to carriers. Even though effective treatment is available the annual number of deaths is rising, indicating that better tolerated treatments are needed. CONCLUSION: A national diagnosis register is a powerful instrument for monitoring the treatment and prevention of inherited disorders and for highlighting correctable shortcomings. In view of the increasing possibilities for genetic screening there is a strong case for central funding for such databases within modern health services. PMID- 11731808 TI - Malaria control in Bungoma District, Kenya: a survey of home treatment of children with fever, bednet use and attendance at antenatal clinics. AB - OBJECTIVE: To lay the basis for planning an improved malaria control programme in Bungoma District, Kenya. METHODS: By means of a cluster sample household survey an investigation was conducted into the home management of febrile children, the use of bednets, and attendance at antenatal clinics. FINDINGS: Female carers provided information on 314 recently febrile children under 5 years of age, of whom 43% received care at a health facility, 47% received an antimalarial drug at home, and 25% received neither. Of the antimalarial treatments given at home, 91% were started by the second day of fever and 92% were with chloroquine, the nationally recommended antimalarial at the time. The recommended dosage of chloroquine to be administered over three days was 25 mg/kg but the median chloroquine tablet or syrup dosage given over the first three days of treatment was 15 mg/kg. The total dosages ranged from 2.5 mg/kg to 82 mg/kg, administered over one to five days. The dosages were lower when syrup was administered than when tablets were used. Only 5% of children under 5 years of age slept under a bednet. No bednets had been treated with insecticide since purchase. At least two antenatal visits were made by 91% of pregnant women. CONCLUSIONS: Carers are major and prompt providers of antimalarial treatment. Home treatment practices should be strengthened and endorsed when prompt treatment at a health facility is impossible. The administration of incorrect dosages, which proved common with chloroquine, may occur less frequently with sulfadoxine-pyrimethamine, as its dosage regimen is simpler. High levels of utilization of antenatal clinics afford the opportunity to achieve good coverage with presumptive intermittent malaria treatments during pregnancy, and to reach the goal of widespread bednet use by pregnant women and children by distributing nets during antenatal clinic visits. PMID- 11731809 TI - Comparison of two training strategies for essential newborn care in Brazil. AB - OBJECTIVE: To compare the effectiveness of two training strategies for improving essential newborn care in the state of Pernambuco, Brazil. METHODS: Eight hospitals were selected, divided into two groups of four, and paired by geographical, structural, and functional characteristics. Doctors and nurses working at hospitals in Group 1 were given a conventional 5-day training course. Those in Group 2 were given the same manual used by Group 1 but the training course was organized as self-directed learning, with the participants having 5 weeks to complete the course. Participants' knowledge was tested at baseline, immediately after the course, and 3-6 months later. Participants' practices were observed before training and 3-6 months after training during 20 births and by interviewing 20 mothers before discharge at each hospital. FINDINGS: Not all participants completed all of the tests. The scores on the tests of knowledge improved more among those in Group 2 than those in Group 1 when the answers were classified as right or wrong, but there was no difference between groups when a scoring method was used that classified answers as correct, partially correct, incorrect, or missing. Practices related to thermal control after birth improved among those in Group 2 after training but practices related to thermal control on the ward worsened. The promotion of breastfeeding improved in both groups. CONCLUSION: There was no difference between the two training strategies, although self-directed learning was cheaper than conventional training. Neither strategy brought about the expected improvements in the quality of care. Other interventions in addition to training may be needed to improve care. PMID- 11731810 TI - User fees, health staff incentives, and service utilization in Kabarole District, Uganda. AB - OBJECTIVE: To determine the impact of user fees on the utilization of health services in a community-based cost-sharing scheme in Kabarole District, western Uganda. METHODS: Of the 38 government health units that had introduced user-fee financing schemes, 11 were included in the study. Outpatient utilization was assessed as the median number of visits per month before and after cost sharing began. FINDINGS: After the introduction of cost sharing, overall utilization of general outpatient services, assessed by combining the data from all the participating units, dropped by 21.3%. Utilization increased, however, in facilities located in remote areas, while it decreased in those located in urban or semi-urban areas. The increased utilization in remote facilities was considered to be largely attributable to health workers' incentive payments derived from cost-sharing revenues. CONCLUSIONS: Incentive payments led the health workers to offer improved services. Other factors may also have been influential, such as an improved drug supply to health facilities and increased public identification with community projects in remote areas. PMID- 11731811 TI - Immune response to measles vaccine in Peruvian children. AB - OBJECTIVE: To evaluate the immune response in Peruvian children following measles vaccination. METHODS: Fifty-five Peruvian children received Schwarz measles vaccine (about 10(3) plaque forming units) at about 9 months of age. Blood samples were taken before vaccination, then twice after vaccination: one sample at between 1 and 4 weeks after vaccination and the final sample 3 months post vaccination for evaluation of immune cell phenotype and lymphoproliferative responses to measles and non-measles antigens. Measles-specific antibodies were measured by plaque reduction neutralization. FINDINGS: The humoral response developed rapidly after vaccination; only 4 of the 55 children (7%) had plaque reduction neutralization titres <200 mlU/ml 3 months after vaccination. However, only 8 out of 35 children tested (23%) had lymphoproliferative responses to measles antigens 3-4 weeks after vaccination. Children with poor lymphoproliferative responses to measles antigens had readily detectable lymphoproliferative responses to other antigens. Flow cytometric analysis of peripheral blood mononuclear cells revealed diffuse immune system activation at the time of vaccination in most children. The capacity to mount a lymphoproliferative response to measles antigens was associated with expression of CD45RO on CD4+ T-cells. CONCLUSION: The 55 Peruvian children had excellent antibody responses after measles vaccination, but only 23% (8 out of 35) generated detectable lymphoproliferative responses to measles antigens (compared with 55-67% in children in the industrialized world). This difference may contribute to the less than uniform success of measles vaccination programmes in the developing world. PMID- 11731812 TI - Prevalence of impairments, disabilities, handicaps and quality of life in the general population: a review of recent literature. AB - OBJECTIVE: To determine the prevalence rates of morbidity in the general population through bibliographic research. METHODS: Articles relating to impairment, disability, handicap, quality of life and their prevalence in the general population, published between January 1990 and March 1998, were selected on the MEDLINE database. FINDINGS: The 20 articles retained out of 433 used 41 different indicators. Indicators of impairment, disability, handicap and low quality of life showed prevalence rates of 0.1-92%, 3.6-66%, 0.6-56% and 1.8-26% respectively, depending on age and the accuracy of indicators. The heterogeneity of the conceptual framework and insufficient recognition of the importance of indicator accuracy, the age factor and the socioeconomic characteristics of the studied populations impede reliable international comparison. CONCLUSION: Further standardization of indicators is therefore required. The revision of the International Classification of Impairments, Disabilities and Handicaps may make it possible to resolve some of the difficulties encountered. PMID- 11731813 TI - Antibody response to 17D yellow fever vaccine in Ghanaian infants. AB - OBJECTIVES: To assess the seroresponses to yellow fever vaccination at 6 and 9 months of age; assess any possible adverse effects of immunization with the 17D yellow fever vaccine in infants, particularly at 6 months of age. METHODS: Four hundred and twenty infants who had completed BCG, OPV and DPT immunizations were randomized to receive yellow fever immunization at either 6 or 9 months. A single dose of 0.5 ml of the reconstituted vaccine was administered to each infant by subcutaneous injection. To determine the yellow fever antibody levels of the infants, each donated 1 ml whole blood prior to immunization and 3 months post immunization. Each serum sample was titred on Vero cells against the vaccine virus. FINDINGS: The most common adverse reactions reported were fever, cough, diarrhoea and mild reactions at the inoculation site. The incidences of adverse reactions were not statistically different in both groups. None of the pre immunization sera in both age groups had detectable yellow fever antibodies. Infants immunized at 6 months recorded seroconversion of 98.6% and those immunized at 9 months recorded 98% seroconversion. The GMT of their antibodies were 158.5 and 129.8, respectively. CONCLUSIONS: The results indicate that seroresponses to yellow fever immunization at 6 and 9 months as determined by seroconversion and GMTs of antibodies are similar. The findings of good seroresponses at 6 months without significant adverse effects would suggest that the 17D yellow fever vaccine could be recommended for use in children at 6 months in outbreak situations or in high risk endemic areas. PMID- 11731814 TI - Malathion resistance and prevalence of the malathion carboxylesterase mechanism in populations of mosquito vectors of disease in Sri Lanka. AB - OBJECTIVE: To determine the levels of malathion resistance and prevalence of the malathion carboxylesterase mechanism among mosquitoes in Sri Lanka. METHODS: Bioassays were carried out using WHO-recommended methods on samples of the following Sri Lankan mosquito vectors: Culex quinquefasciatus, C. tritaeniorhynchus, C gelidus, Anopheles culicifacies B, A. subpictus, Aedes aegypti and A. albopictus. FINDINGS Malathion-specific carboxylesterase mechanisms were found in A. culicifaies and A. subpictus, both giving high rates of insecticide metabolism. In contrast, malathion resistance in C. quinquefasciatus and C. tritaeniorhynchus is linked to broad-spectrum resistance to organophosphorus compounds due to elevated levels of esterases that sequester malaoxon, but are unable to metabolize malathion. CONCLUSIONS: Resistance among the Anophelesspp. must have occurred as a direct result of antimalarial activities, since malathion use in Sri Lanka is limited to public health treatments. In contrast, resistance among Culex spp. has resulted from large scale use of the organophosphorus insecticide group as larvicides for filariasis control and on rice paddy, where C tritaeniorhynchus predominantly breeds, for agricultural purposes. PMID- 11731815 TI - The contribution of reproductive ill-health to the overall burden of perceived illness among women in southern India. AB - OBJECTIVE: To investigate women's perceptions of the overall burden of illness among a sample of women in southern India. METHODS: A community-based sample of 421 young married women in a subdistrict about 70 kilometres from Bangalore, Karnataka State, India, were interviewed monthly for one year. At each visit, information on the symptoms of all forms of illness they had experienced was elicited with the aid of a checklist. Details were obtained on the durations of episodes of illness and on health-seeking behaviour and costs. The symptoms were subsequently coded in accordance with the International Classification of Diseases (ICD-10). FINDINGS: Reproductive ill-health accounted for half of all illness-days and for 31% of total curative health expenditure. The 1990 Global Burden of Disease study estimated that 27.4% of disability-adjusted life years (DALYs) lost in Indian women aged 15-44 years were attributable to reproductive ill-health. CONCLUSIONS: Our study indicates that this dimension of morbidity, when measured in terms of women's subjective experiences, makes a larger contribution to the burden of illness than that suggested by the DALY approach. This lends justification to the high priority attached to reproductive ill-health in India. PMID- 11731816 TI - Syndromic treatment of gonococcal and chlamydial infections in women seeking primary care for the genital discharge syndrome: decision-making. AB - The syndromic treatment of gonococcal and chlamydial infections in women seeking primary care in clinics where resources are scarce, as recommended by WHO and implemented in many developing countries, necessitates a balance to be struck between overtreatment and undertreatment. The present paper identifies factors that are relevant to the selection of specific strategies for syndromic treatment in the above circumstances. Among them are the general aspects of decision-making and caveats concerning the rational decision-making approach. The positive and negative implications are outlined of providing or withholding treatment following a specific algorithm with a given accuracy to detect infection, i.e. sensitivity, specificity and predictive values. Other decision-making considerations that are identified are related to implementation and include the stability of risk factors with regard to time, space and the implementer, acceptability by stakeholders, and environmental constraints. There is a need to consider empirically developed treatment algorithms as a basis for policy discourse, to be evaluated together with the evidence, alternatives and arguments by the stakeholders. PMID- 11731817 TI - The burden of disease and injury in Australia. AB - An overview of the results of the Australian Burden of Disease (ABD) study is presented. The ABD study was the first to use methodology developed for the Global Burden of Disease study to measure the burden of disease and injury in a developed country. In 1996, mental disorders were the main causes of disability burden, responsible for nearly 30% of total years of life lost to disability (YLD), with depression accounting for 8% of the total YLD. Ischaemic heart disease and stroke were the main contributors to the disease burden disability adjusted life years (DALYs), together causing nearly 18% of the total disease burden. Risk factors such as smoking, alcohol consumption, physical inactivity, hypertension, high blood cholesterol, obesity and inadequate fruit and vegetable consumption were responsible for much of the overall disease burden in Australia. The lessons learnt from the ABD study are discussed, together with methodological issues that require further attention. PMID- 11731818 TI - Tobacco control in tobacco-producing countries. PMID- 11731819 TI - Continent-wide attack launched on African trypanosomiasis. PMID- 11731821 TI - WHO helps countries prepare for bioterror attacks. PMID- 11731820 TI - For Afghan polio eradication the show goes on. PMID- 11731822 TI - WHO finds governments "remiss" on mental disorders. PMID- 11731823 TI - Planet Earth, getting too hot for health? PMID- 11731824 TI - Dissolution of lead pellets in saliva: a source of lead exposure in children. PMID- 11731825 TI - Effects on growth and changes in organ distribution of bullfrog larvae exposed to lead throughout metamorphosis. PMID- 11731826 TI - Effect of oral exposure to cadmium on the levels of vitamin A and lipid peroxidation in the eye. PMID- 11731827 TI - Biological indices in formulators exposed to a combination of pesticides. PMID- 11731828 TI - Toxicity of stormwater runoff after dormant spray application of diazinon and esfenvalerate (Asana) in a French prune orchard, Glenn county, California, USA. PMID- 11731829 TI - Size dependent tolerance of Indian cat fish Heteropneustes fossilis (Bloch) to toxicity of cadmium and composted vegetation. PMID- 11731830 TI - Effects of metal exposures on juvenile clams, Mercenaria mercenaria. PMID- 11731831 TI - Copper and manganese influence the uptake of cadmium in marine macroalgae. PMID- 11731832 TI - Growth of flax plants (Linum usitatissimum) as affected by water and sludge from a sewage treatment plant. PMID- 11731833 TI - Foot transfer of lawn-applied pesticides from turf to carpet: comparison of semivolatile chlorpyrifos with nonvolatile chlorothalonil. PMID- 11731834 TI - Pesticides and polychlorinated biphenyl residues in human adipose tissue. PMID- 11731835 TI - Development of plant bioassay to detect herbicide contamination of compost at or below practical analytical detection limits. PMID- 11731836 TI - Pesticide residues in soil from the Jordan Valley, Jordan. PMID- 11731837 TI - Persistent organochlorine pollutants in soils from belgium, Italy, Greece, and Romania. PMID- 11731838 TI - Polycyclic aromatic hydrocarbons (PAHs) in the sediments and fish of the Mill River, New Haven, Connecticut, USA. PMID- 11731839 TI - Measuring TCDD equivalents in environmental samples with the micro-EROD assay: comparison with HRGC/HRMS data. PMID- 11731840 TI - Influence of leaching protocol regimes on losses of wood preservative biocides. PMID- 11731841 TI - 4-nonylphenol, 4-nonylphenol mono- and diethoxylates, and other 4-alkylphenols in water and shellfish from rivers flowing into lake Biwa. PMID- 11731842 TI - Arsenic and cadmium levels in imported fresh and frozen fish in Jordan. PMID- 11731843 TI - Levels of reactive mercury and silver in sediments from the port of Ensenada, Baja California, Mexico. PMID- 11731844 TI - Polycyclic aromatic hydrocarbons in leachate from a municipal solid waste dump of Niteroi City, RJ, Brazil. PMID- 11731845 TI - Mammary augmentation by cohesive silicone gel implants with anatomic shape: technical considerations. AB - Anatomically formed cohesive silicon gel implants for mammary augmentation or restoration were first introduced in 1993. They have been allowed for use in the most of European countries, but not in the Northern America for the past few years. These implants are appreciated by many patients, but they are still controversial, because the whole concept of treatment is different from the techniques used for implants filled with normal saline or liquid silicone. Cohesive gel implants can give us excellent results, but the margin for mistakes is near zero. Information about the new techniques necessary to use with these implants has so far been scarce. This paper gives detailed description of the preoperative preparations, the operation step-by-step, and the postoperative care for patients with the cohesive gel implants and is based on the four years experience with 440 patients. Our personal technique using the intracutaneous wound closure with totally buried resorbable suture and the implant of new design, with the marking string on the front and fixation plate on the posterior surface, is presented. PMID- 11731846 TI - Circumareolar dermo-glandular plication: a new concept for correction of breast ptosis. AB - Circumareolar dermo-glandular plication is the latest advancement of the periareolar dermopexy with a retromammary mastopexy technique I published in 1969. Rather than a technique, the new concept is a procedure which originates new techniques covering multiple indications, i.e. for all conditions combined with ptosis: for mastopexy in ptotic breasts, for hypertrophic or hypoplastic breasts with resection or implant augmentation, respectively; for subcutaneous mastectomy, gynecomastia, asymmetries, and tuberous breasts. It is useful for reoperations to correct secondary ptosis as well as to reduce the length of the scar in vertical techniques. The corresponding techniques are described. The procedure has proved to be safe and reliable in over 200 patients with the following advantages: no full thickness skin incision or excisions are performed; only the epidermis is excised. Except for hypertrophies, the skin is not dissected from the gland, nor the gland from the pectoralis fascia, which increases vascular safety and preserves NAC innervation; the dermoglandular unit of the breast through Cooper's ligaments is stabilized by a single or multiple plications. The scar is only circumareolar, reducing psychological stress and discomfort and achieving an early recovery and patient satisfaction. The inconveniences are puckering and some widening of the periareolar scar, which requires a secondary revision in approximately 50% of the cases, also frequently necessary in conventional techniques. There is a tendency to flattening of the NAC and periareolar bulging with tendency to a "tomato breast appearance." The prevention of the latter is described. PMID- 11731847 TI - Prospective study of clinical findings and changes in 56 Trilucent implant explantations. AB - Between 1995 and 1998 we implanted 88 Trilucent implants in 48 patients. The experience of 56 explantations in 30 patients are presented in this prospective study. Of 48 patients, 32 returned for review after we wrote to them. Twenty seven elected to have their implants exchanged immediately for a fourth generation cohesive silicon implant and three decided to have the implants removed and not replaced. In 14 patients it was clinically obvious that the volume of the implant had changed, although not all patients realized this. The absence of capsular contraction was notable (unanimously Baker II), so that most patients were asymptomatic and had to be convinced of the need for explantation. However, perioperatively, 55% of the implants had thickening or color change caused by the peroxidation of the triglyceride content. Typically the implant capsule was adherent to the surrounding tissues, especially pectoralis major. This prolonged operative time (184 min, on average) and hemostasis was a problem. During the study we developed a standardized operative technique, which enabled us to reduce operative times. Special attention had to be paid to the selection of the new implant volume, because many patients had become accustomed to the increase in the size of their breasts caused by the peroxidation of the implant content. Forty-three percent of patients preoperatively expressed the wish to have even bigger breasts than before. Nearly all of our patients at the three month postoperative follow-up were happier with the new implants than before. It became apparent that after only two to three years there were obvious oxidative changes in the implants in asymptomatic patients. Based on our study result, the recommendations regarding explantation of Trilucent implants seem justified. PMID- 11731848 TI - A different strategy in the surgical treatment of capsular contracture: leave capsule intact. AB - The authors present their experience with the surgical treatment of capsular contracture to achieve better results in a safe, predictable, and practical way, and discuss the possible treatment modalities. They simply advise leaving the capsule intact, even if it is calcified, and create another pocket, rarely in the front or, more typically, at the back of the capsule. If the breast tissue is also ptotic, a mastopexy procedure may be added to the procedure, in addition to augmentation, with a rather small prosthesis placed in the new pocket or, occasionally, in the old one. External, forceable massage is not advisable to treat the capsule. Open capsulotomy and/or partial capsulectomy can be applied to release the capsule. However, it is not advisable since recurrence is usually inevitable. The purpose of this paper is to present a series of surgical procedures to avoid the problems created by the capsule and present different cases with good results. PMID- 11731849 TI - Specimen radiography: an assessment method for reduction mammaplasty materials. AB - Although, one out of every eight women has a risk of developing breast cancer, the reported incidence of breast carcinoma detection in reduction mammaplasty materials is rather low. To our knowledge, specimen radiography, which is used for breast biopsies has not been used for the assessment of breast reduction materials. We investigated the applicability of specimen radiography and its potential benefits in detection of the breast pathologies, especially malignancies in reduction mammaplasty materials. Forty patients scheduled for reduction mammaplasty operation were included. In all cases an inferior pedicle reduction technique was preferred and the radiographs of the resected breast tissues were taken immediately. The radiographs were evaluated for any possible pathologic appearance and all abnormal findings were marked. For the histopathologic evaluation, in addition to the random sampling of the pathologist, any marked areas were also microscopically examined. In two cases fibrocystic changes were found in radiographs and the same results were obtained in the histological examination. No false negative mammogram was seen. Specimen radiography, which is applicable for breast reduction materials is an easy and cheap method and does not cause any patient discomfort. It seems that the radiographs of reduction mammaplasty materials are useful to provide guidance to the pathologist during tissue sampling for microscopic examination especially when large amounts of breast tissue is excised. PMID- 11731850 TI - Innovative new concepts in augmentative breast surgery. Part II: systematic and drawing. AB - This paper presents a simple protocol to define the anatomic parameters for creating aesthetic augmented breasts using round silicone implants. This study completes our previous publication on augmentative breast surgery. We discuss the need for correct submuscular dissection and explain how to obtain the correct positioning of the inframammary fold via measurements. We also explain how we use radiological images to show the continuity of the submuscular dissection in the inferior border so that we can create a new submammary fold and thus preserve the central position of the nipple-areola complex. PMID- 11731851 TI - An analysis of breast sensation following inferior pedicle mammaplasty and the effect of the volume of resected tissue. AB - The aim of our study was to evaluate if the amount of resected breast tissue has an influence on breast sensitivity after inferior pedicle mammaplasty. In the prospective study, 15 patients (30 breasts) were divided into two groups: group I (less than 400 g resection) and group II (more than 400 g resection). Preoperatively and six months postoperatively a touch test (Semmes-Weinstein monofilaments) and temperature (warm and cold) tests were performed. The study showed that all patients had increased touch sensitivity six months after inferior pedicle mammaplasty regardless of the amount of resection. In group I an average of 65% and in group II an average of 83% retained sensitivity for temperature postoperatively. These results suggests that this mammaplasty technique preserves intercostal nerves within the inferior pedicle and reliefs chronic nerve traction injury and improves thereby breast sensibility. PMID- 11731852 TI - Minimally invasive facial rejuvenation endolaser mid-face lift. AB - Endolaser mid-face lift was performed on patients in a multi-center study over a 36-month period (Feb. 1998 to Feb. 2001). It permits significant facial rejuvenation through small incisions. This technique achieves aesthetic results and wider rejuvenation while being less traumatic and creating minimal morbidity. Combined with other procedures, it rejuvenates the face by three strategic methods: soft tissue suspension, reversal of photo aging, and correction of the depletion of volume. To achieve this triple result, the mid-face lift is performed by endoscopic approach, and in every case is combined with the endoscopic lift of the frontal area. Laser resurfacing was used to reverse skin photo damage. The Ultrapulse CO2 laser and/or the Ultrafine Erbium YAG(Coherent, Inc, Palo Alto, CA) were used. The third combined procedure was the introduction of fat graft to compensate the atrophy/ptosis of fat and the depletion of bone mass (other filling materials besides fat may be used, depending on the preference of the surgeon). Our method of fixation using the Casagrande Needle (an evolution of Reverdin Needle) makes the mechanical purchase on the tissues to be suspended much easier, permitting the intra-oral and/or infra-orbital incisions to be eliminated. The present study of the technical evolution of the endolaser mid-face lift method allows us to conclude that a very satisfactory outcome has been reached, offering patients a minimally invasive procedure, which can be performed under local anesthesia, with low morbidity, imperceptible incisions, and an excellent long-term result. PMID- 11731853 TI - Transparent dressing for rhytidectomy. AB - Several dressings have been proposed for rhytidectomy. At present, the type most frequently used consists of flat cotton strips soaked in mineral oil placed on the incisions and along the mandibular line. Gauze sponges are then placed on the cheeks and an elastic net is wrapped around the head and left open anteriorly to expose the face and eyes. Despite its widespread use, this technique presents problems. It is uncomfortable for the patient, it is tight and confining, it obstructs hearing, complicates hygiene, attracts unnecessary attention, and, most importantly, does not permit visualization of the surgical site or of the complications that might arise, such as hematomas. Thus, we propose a more efficient dressing made of sterilized, transparent PVC film of the type used to protect food in the refrigerator (Saran Wrap-like). This dressing has proved to be more comfortable for the patient, is not tight or confining, does not cause changes in hearing, facilitates local hygiene, does not attract unnecessary attention on the part of the patient himself or of third parties and, most importantly, it is transparent and adheres to all points of the skin flap, permitting early visualization of complications, such as hematomas. PMID- 11731854 TI - Multiple atrophies following tooth extraction. AB - Following a molar tooth extraction, the patient developed a depression and atrophy of the right ear, the right half of the mentum, and the tongue. These atrophy deformities were successfully corrected with dermografts. PMID- 11731855 TI - Tissue paper for hemostasis of spot dermabrasion. AB - Dermabrasion remains one of the effective ways to treat scars and facial wrinkles. Spot dermabrasion is for a localized limited area. Bleeding and oozing from the abraded wound are cumbersome during the operation. Small pieces of tissue paper were used for the intraoperative hemostasis. The efficiency and safety of the tissue paper as a temporary dressing for hemostasis was discussed and presented. It is simple, cost-effective, and easy to use for hemostasis from spot dermabrasion. PMID- 11731856 TI - Endoscopic balloon dissection for removal of lipomas via transaxillary route. AB - Minimally invasive techniques and newer endoscopic surgical techniques are being introduced to aesthetic and reconstructive procedures at an accelerating pace. A new and modified technique of endoscopic excision of large encapsulated lipomas is presented and the versatility of creation of an optical cavity and incorporation of balloon dissection is discussed in two cases. Large encapsulated lipomas were removed endoscopically via transaxillary incisions and no immediate or late complication was observed. Endoscopic removal of lipomas offers several advantages over conventional direct excision and liposuction such as avoiding blind manipulation and fragmentation of lipoma mass, and unsightly scars. One drawback of this technique can be lengthy operation times, which have been near two hours for the presented cases. We conclude that endoscopic removal of tumors from a relatively remote site is beneficial in treating such encapsulated tumors and we speculate that endoscopy will prevent unsightly scarring in important areas while decreasing the operating time as experience is gained. PMID- 11731857 TI - The skin of the medial area of the arm: morphometric study of interest to liposuction. AB - The study of the skin of medial area of the arm in cadavers made it possible to evaluate its regional characteristics. Sex or race were not reported, but the age group most appropriate to liposuction was considered. In this way, authors believed that, as per standards established at the time, results as to skin retraction in liposuction, will be able to be analyzed in a comparative way in the future. PMID- 11731858 TI - Psychopathology and body image in cosmetic surgery patients. AB - The purpose of this study was to investigate the presence of psychiatric symptoms and evaluate the perceptive, cognitive, and behavioral aspects of body image in cosmetic surgery patients. These parameters of 20 cosmetic patients and of 20 control patients matched for age, gender, education, and marital status who attended the general surgery department for minor surgery were compared in a cross-sectional design. Symptom Check List-90 (SCL-90), Beck Depression Inventory (BDI), and The Multi-Dimensional Body Self Relations Questionnaire were administered to both groups. No significant difference was determined in the rates of psychopathology of the patient and control groups. Scales assessing self image did not indicate any significant difference between the groups. Four (20%) of the cosmetic patients, however, were diagnosed with body dysmorphic disorder according to DSM-IV. Cosmetic patients were usually defensive towards psychological evaluations. A wide range of diversity was determined in the psychiatric evaluation of the cosmetic patient group. While some patients exhibited healthy psychological traits, some had severe depressive disorder or nearly psychotic somatic preoccupations. PMID- 11731859 TI - Liposuction and scar placement. AB - A simple methodology is described that addresses liposuction procedures and the placement of scars. Natural body creases are employed to disguise scarring. PMID- 11731860 TI - Virtual MRI endoscopy: detection of anomalies of the ventricular anatomy and its possible role as a presurgical planning tool for endoscopic third ventriculostomy. AB - BACKGROUND: Many anatomical anomalies have the potential to impair the efficacy of endoscopic third ventriculostomy (ETV) and increase the surgical morbidity. By virtual magnetic resonance imaging (MRI) endoscopy, the real endoscopic view into the ventricular system can be simulated. It was the objective of the present study to investigate if this simulation is sensitive enough to detect anatomical anomalies of the ventricular system. METHOD: In 18 hydrocephalic patients, first neuronavigationally guided ETV, then virtual MRI endoscopy were performed. This study design allowed for selection of a path for virtual MRI endoscopy, which was identical to that used during surgery, making the real and the virtual view on anatomical structures of the ventricular system highly comparable. It was investigated whether the intra-operatively identified anatomical anomalies could likewise be depicted on virtual MR endoscopic images. FINDINGS: Seven anatomical variants (not enlarged interventricular foramen n=2, atrophic corpus callosum and split fornical bodies n=1, narrow retroclival space n=1, prominent basilar tip n=1, opaque and thick/atypically declining third ventricular floor n=2) were encountered in 5 of the 18 patients during surgery. The five variants of the non membraneous structures were identified by virtual MRI endoscopy (sensitivity 71%), whereas the anatomical variants of the third ventricular floor were missed. Both the normal as well as the variant third ventricular floor could not be visualised and appeared as a defect. Through this artefact, the anatomy of the major vessels in the interpeduncular cistern could be assessed. INTERPRETATION: The sensitivity of virtual MRI endoscopy for detection of anatomical variants of the ventricular system is low. Its potential usefulness as a presurgical planning tool inspite of this low sensitivity rate is discussed. PMID- 11731861 TI - The management of residual and recurrent intracranial aneurysms after previous endovascular or surgical treatment--a report of eighteen cases. AB - OBJECT: We wish to report our experience in the management of residual or recurrent intracranial aneurysm after previous endovascular or surgical treatment. METHODS: We performed a retrospective review of the clinical notes, operation records and cerebral angiograms of eighteen patients who were known to have undergone treatment for residual or recurrent aneurysms. RESULTS: During the period of April 1994 to May 1999, 210 patients were treated for an intracranial aneurysm either surgically or by endovascular methods. Eighteen of these patients (8.6%) were subsequently treated for residual or recurrent aneurysm. Thirteen achieved a complete occlusion. Complete occlusion was achieved in five of the eight patients who underwent endovascular treatment as a second procedure. Seven out of ten surgical cases achieved complete occlusion. Fifteen patients made a good recovery according to the Glasgow Outcome Score. Two patients who presented in a poor grade subarachnoid haemorrhage (SAH) were left severely disabled. One patient died after retreatment. CONCLUSIONS: The treatment of cerebral aneurysm remnants can be performed effectively using a variety of modalities. The original purpose of the treatment, which is total occlusion of the lesion, can thus be achieved. PMID- 11731862 TI - Determining optimal MRI follow-up after transsphenoidal surgery for pituitary adenoma: scan at 24 hours postsurgery provides reliable information. AB - There is no agreed-upon schedule for MRI follow-up after pituitary adenoma removal via the transsphenoidal approach. Our aim was to establish a plan for MRI follow-up after pituitary surgery. Eighty pituitary adenoma cases (25 microadenomas, 30 macroadenomas that did not infiltrate the cavernous sinus, and 25 macroadenomas with cavernous sinus infiltration) were prospectively studied with MRI following tumor resection via the transsphenoidal approach. Each patient was imaged at 24 hours, at 3, 6 and 9 months, and at 1 year or more postsurgery. The parameters studied were residual tumor, synthetic packing material (Gelfoam) versus fat graft, and normal pituitary, hypophyseal stalk and optic chiasma. The size of the pituitary structure as a whole was also measured. The latter was studied quantitatively, and the findings for the rest of the parameters were evaluated qualitatively, based on the examiners' confidence in their assessment. The final MRI study, done at least 1 year postsurgery in all cases, was considered the reference MRI for all scans. MRI performed 24 hours after surgery was diagnostically accurate for residual tumor and valuable for visualizing normal sellar structures. The findings also showed that fat packing takes longer to resorb than Gelfoam, but produces no potentially confounding contrast enhancement. An algorithm based on the results is presented for postsurgical MRI assessment of pituitary adenoma patients in which the scan at 24 hours postsurgery is the major factor that determines the timing of later rechecks. PMID- 11731863 TI - Initial experience with an ultrasound-integrated single-RACK neuronavigation system. AB - A prototype ultrasound-integrated neuronavigation system was tested in 34 operations as regards image quality, stability, and handling during daily use in the operating theatre. The system consists of a high-end ultrasound scanner, a navigation computer, and an active optical positioning and digitiser system, all integrated in a single rack. An Ethernet interface between the two hardware devices enables digital data transfer between the ultrasound scanner and the navigation device without loss of image quality. The integration of an ultrasound scanner and a navigation device offers the opportunity of navigating directly to an intracranial or intraspinal lesion using intra-operative 3D ultrasound images. The brainshift problem is thus avoided. The ability to directly compare MR images and 3D ultrasound simplifies the interpretation of ultrasound images. The single rack solution is an advantage in times of restricted space in the operating theatre caused by the increasing volume of technical equipment needed for a neurosurgical operation. In 30 cases the prototype system showed good reliability. In four cases the navigation system failed during the operation; however, the capacity of the ultrasound scanner was still available as a stand alone function. With the single-rack concept, the flexibility of the system is high and the complete device can easily be moved from one operating theatre to another. PMID- 11731864 TI - MRimaging findings after ventricular puncture in patients with SAH. AB - OBJECT: Using magnetic resonance (MR) imaging, we studied brain injury from ventricular puncture performed during craniotomy in the acute stage of subarachnoid hemorrhage (SAH). METHODS: 80 patients underwent craniotomy for aneurysm obliteration within 48 hr after SAH, ventricular puncture for drainage of cerebrospinal fluid (CSF) was performed to reduce intracranial pressure. MR imaging was performed within 3 days following surgery to measure the size of the lesion, and was repeated on postoperative days 14 and 30. CONCLUSIONS: Of the 80 patients with ventricular puncture preceding craniotomy, 65 (81%) showed MR evidence of brain injury from the puncture. Overall, 149 lesions were detected. According to coronal images, cortical injuries (54 cases), penetrating injury to tracts along the ventricular tube (55 cases), caudate injury (25 cases), and corpus callosum injury (15 cases). Brain injuries from ventricular puncture did not correlate significantly to patient outcome. While ventricular puncture and drainage of CSF can readily be performed to decrease brain volume at the time of craniotomy in acute-stage SAH, neurosurgeons should be aware of a surprisingly high incidence of brain injury complicating puncture. PMID- 11731865 TI - Linac radiosurgery for skull base meningiomas. AB - INTRODUCTION: Skull base meningiomas present a difficult surgical challenge because of the high potential morbidity of radical surgical extirpation and their low potential for incapacitating symptomatology. The focal character of meningiomas makes stereotactic radiosurgery an attractive adjuvant treatment modality to resection. The purpose of this study was to evaluate the local control rates and complications in 56 patients with base of skull meningiomas undergoing radiosurgery. METHODS: Patients underwent radiosurgery using the dedicated stereotactic linear accelerator at the Brigham and Women's Hospital. Minimal peripheral doses of radiosurgery ranged from 12 to 18.5 Gy (mean 15 Gy). Doses were designed to conform to the frequently irregular tumor volumes using the X-Knife treatment planning system. Multiple isocenters were used when required to increase conformality of dose. For 36 patients (64%), radiosurgery was used as an adjunct to surgery; for 20 patients (36%) it was the primary treatment. RESULTS: Median followup was five years. Nineteen patients (34%) were improved clinically at follow-up; 32 (57%) were unchanged; and 5 patients (9%) developed new or worsened neurologic deficits. Serial imaging studies after radiosurgery showed a reduction in tumor volume in 23 patients (41%); 30 (54%) showed stable disease; 3 patients (5%) had tumors which increased in size (2 being outside the radiosurgery treatment site). The actuarial freedom from progression rate (defined as further tumor growth) was thus 95%, with a median imaging follow-up of 26 months (range, 6-66 months). Although further follow-up is necessary, the results of this series clearly demonstrate that these lesions are feasible for treatment by modern radiosurgical techniques. Linac radiosurgery can stabilize skull base meningiomas, with decreased or unchanged tumor volumes on radiologic follow-up in approximately 95% of patients. Radiosurgery is a low morbidity, effective technique as adjunct and sometimes primary treatment of small to moderate-sized meningiomas of the skull base. PMID- 11731866 TI - Cerebral aneurysms in the perforating artery manifesting intracerebral and subarachnoid haemorrhage--report of two cases. AB - BACKGROUND: An arteriosclerotic aneurysm in the perforating artery has been focused on as a causative factor for hypertensive intracerebral haemorrhage. However, its pathogenesis remains unknown, and its existence is still a controversy. CASE DESCRIPTION: A 62-year-old female and a 70-year-old male with a history of hypertension suffered from intracerebral haemorrhage accompanied by subarachnoid haemorrhage. Cerebral angiograms demonstrated an aneurysm arising from the perforating artery at the central location of the haematoma in both cases. The aneurysms were confirmed as the cause of bleeding during microsurgery, and were resected. Histological examination of the surgical specimens revealed that the walls of the aneurysms lacked internal elastic lamina and consisted only of the adventitia. CONCLUSION: These findings demonstrate that the aneurysm in the perforating artery can be a causative factor for hypertensive intracerebral haemorrhage, and indicate that the loss of internal elastic lamina induced by hypertension may contribute to the formation of the aneurysm of the perforating artery. PMID- 11731867 TI - Adrenomedullin reduces ischemic brain injury after transient middle cerebral artery occlusion in rats. AB - BACKGROUND: The effect of adrenomedullin, a vasodilatory peptide on transient middle cerebral artery (MCA) occlusion was investigated in rats. METHODS: Transient MCA occlusion for 2 hours was made by using the intra-arterial suture method, followed by reperfusion. FINDINGS: An intravenous infusion of adrenomedullin (1 microg/kg/min) from one hour before ischemia to one hour after ischemia significantly reduced the infarct size and improved neurological deficits (p<0.05), without affecting systemic blood pressure or other physiological parameters. The infarct size was reduced with adrenomedullin by 25.4+/-12.7%, 31.3+/-5.8%, 31.6+/-6.1% respectively at the coronal level 6, 8 and 10 mm posterior from the frontal pole. Adrenomedullin also significantly inhibited the increase in myeloperoxidase (MPO) activity in the MCA area of the ischemic hemisphere after 22-hour reperfusion (control: 0.205+/-0.054 unit/g wet tissue, adrenomedullin group: 0.047+/-0.009 unit/g wet tissue, p<0.0001). INTERPRETATION: These data suggest that adrenomedullin reduces acute ischemic brain injury and one of is neuroprotective mechanisms may be derived from inhibition of the infiltration of neutrophils into the ischemic tissue. PMID- 11731868 TI - A new idea for a protective balloon system during carotid stenting. AB - Percutaneous transluminal angioplasty with stenting for high-grade carotid stenosis has been recently come into use. However, distal embolic events remain a problem with this procedure compared to results with established carotid endarterectomy. To counteract such problems, various blocking balloon system such as a simple distal blocking balloon system or a thrombi catching system have been used in some instances. This time, a double-balloon system was used as a proximal blocking system during predilation, and an existing distal blocking system was used during practical carotid stenting. These systems were applied to six cases. In addition, we used a modified conventional blood transfusion system for filtration and retrieval of the aspirated blood. Compared with the simple distal blocking balloon system, occurrence of distal emboli could be reduced with our new combined method although the number of cases is too small to reach any definite conclusions. PMID- 11731869 TI - Trapped third ventricle. AB - The authors report the case of a 29-year-old female who presented with symptoms of shunt dysfunction 11 years after first being shunted for an aqueductal stenosis. After numerous revisions she developed an isolated third ventricle, necessitating triventricular shunting to obtain a new equilibrium. An isolated third ventricle is a very rare phenomenon, usually seen in very complex hydrocephalus and only reported on twice before (Filler et al., 1995). Etiological factors postulated in the development of an isolated lateral or fourth ventricle, all seem to contribute also to the development of an isolated third ventricle. PMID- 11731871 TI - An angiographic lesion mimicking pseudo-aneurysm in cerebral arteriovenous malformation. AB - In cerebral arteriovenous malformations (AVMs), a pseudo-aneurysm represents rupture site, and its presence is known as a factor for rebleeding. We report a case of cerebral AVM presenting with intracerebral haemorrhage in which cerebral angiography showed a lesion mimicking pseudo-aneurysm. Although the patient needed urgent surgical decompression, it was delayed because early haematoma evacuation would induce rebleeding from the rupture site. The authors attempted to occlude the pseudoaneurysm interventionally before surgery. After surgical excision, the lesion that was believed to be a pseudo-aneurysm was revealed as a partially thrombosed venous sac having a thick fibrous wall. In this report, the authors discuss the pitfalls in the interpretation of pseudo-aneurysms in angiographic AVM architecture. PMID- 11731870 TI - Trigeminal neuralgia associated with achondroplasia. Case report with literature review. AB - A 59-year-old male with a history of 2 years of typical trigeminal neuralgia manifested the characteristics of achondroplasia. X-ray and magnetic resonance imaging demonstrated basilar impression, deformity of the posterior fossa with marked asymmetry of petrous bone and compression of pons and trigeminal nerve by the left vertebral artery and anterior inferior cerebellar artery. Microvascular decompression was performed through a suboccipital craniectomy. The neuralgia disappeared soon after surgery and remains completely resolved until today. This is the first reported case of trigeminal neuralgia in a patient with achondroplasia. The deformity of the skull base was considered to influence the development of the trigeminal neuralgia. PMID- 11731872 TI - Venous cerebral infarction due to simultaneous occurrence of dural arteriovenous fistula and developmental venous anomaly. AB - A case of a left frontal lobe infarction in a 31-year-old male patient is presented. This patient had bilateral frontal dural arteriovenous fistulae (DAVF) and a left frontal developmental venous anomaly (DVA). It is suggested that the simultaneous occurrence of these vascular anomalies was the cause of his infarction. PMID- 11731873 TI - The value of cortical stimulation applied to the surgery of malignant gliomas in language areas. PMID- 11731874 TI - The complex clinical and genetic classification of inherited ataxias. II. Autosomal recessive ataxias. AB - Autosomal recessive ataxias are a heterogeneous group of rare neurodegenerative diseases characterized by early onset cerebellar ataxia associated with various neurologic, ophthalmologic and systemic signs. In comparison with autosomal dominant ataxias, the group of recessive ataxias is less extensively characterized. In fact, only a few conditions have been genetically characterized. The pathogenesis of these forms is associated with a "loss of function" of specific cellular proteins involved in metabolic homeostasis, cell cycle, and DNA repair/protection processing. The two most common autosomal recessive ataxias, in European countries, are Friedreich's ataxia and ataxia telangiectasia. Other forms are much less frequent, and include ataxia with vitamin E deficiency, abetalipoproteinemia. Refsum's disease, spastic ataxia, infantile onset spinocerebellar ataxia, and ataxia with oculomotor apraxia. These pathological conditions, although extremely rare, have nevertheless to be carefully considered in differential diagnosis, not only for correct nosographical classification, but particularly, for specific prognostic and therapeutic implications. Some of these diseases exhibit a peculiar regional distribution. An updated review of the clinical, genetic, and pathogenic aspects of recessive ataxias is presented. Specific management problems with respect to diagnosis and genetic counseling are discussed. PMID- 11731875 TI - Modification of cortical excitability induced by gabapentin: a study by transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) was employed before and after a single dose of gabapentin to evaluate how this drug affects the activity of excitatory and inhibitory circuits within the motor cortex. Eleven healthy volunteers were studied. For the evaluation of cortical excitability, the following parameters were taken into account: resting and active motor threshold (RMT, AMT); cortical silent period (CSP); and intracortical inhibition (ICI) and facilitation (ICF). Peripheral silent period (PSP) was also detected. All parameters were measured before and 3 and 24 hours after 800 mg gabapentin was administered in a single oral dose. Gabapentin deepened the ICI and suppressed the ICF at 3 h but not at 5 h after dosing. We conclude that, in the normal human brain, gabapentin may act on intracortical excitability by shifting the balance towards less excitation and more inhibition. PMID- 11731876 TI - Adult brain low-grade astrocytomas: survival after surgery and radiotherapy. AB - In order to identify prognostic factors of survival, twelve elements of disease and treatment have been evaluated for a population of 49 patients with diffuse low-grade astrocytoma treated with surgical resection and radiotherapy. The survival values were inversely correlated with age and major residual portion. On the other hand, KPS, lobar site, grade II Daumas-Duport lesions, protoplasmatic variant, early epilepsy, hyperfractionated radiotherapy and extent of exeresis were prognostic factors correlated with survival. Tumor extent and radiation total dose were not correlated in a meaningful way. Only KPS was statistically significant when compared to all the prognostic factors. We believe that patient selection according to age, lesion site and histological features are not sufficient to generate a homogeneous tumoral population. The most appropriate therapy for treating low-grade astrocytomas is still an open subject. However, recent studies have shown that the prognostic value of a group of factors is useful to plan controlled studies that compare differentiated treatment protocols. PMID- 11731877 TI - Health-related quality of life in type 1 diabetic patients and influence of peripheral nerve involvement. AB - We examined the relationships between diabetic peripheral nerve involvement and patients' own perception of quality of life in a multidimensional study based on outcome research recommendations. The study enrolled 50 consecutive outpatients with type 1 diabetes mellitus (23 men, 27 women; mean age, 40.1 years). The patients were evaluated with validated clinical tests (e. g. Semmes-Weinstein, vibration perception threshold, muscle strength), neurophysiological tests (sural, peroneal and ulnar nerves) and patient-oriented (SF-36 and NASS) questionnaires. Patient-oriented physical scores were significantly related with: (1) neurophysiological findings of the lower limbs; (2) conventional measurements of sensitivity. Conversely, patient-oriented mental scores were not related with these findings. The patient-oriented questionnaires provided an important perspective of the severity of the disease, often closely related with the biological parameters. They offered new interpretations to conventional biological measurements. In particular, the peripheral nerve picture was strictly related to the physical aspects of the patients' quality of life, and not with the mental aspects. PMID- 11731878 TI - Comparison of natural histories of progressive supranuclear palsy and multiple system atrophy. AB - In order to identify early clinical features and survival predictors of supranuclear palsy (PSP) and multiple system atrophy (MSA), we compared the disease course of patients consecutively referred between 1987 and 1999 and followed to December 1999. Thirty-nine PSP and 74 MSA patients were diagnosed according to commonly accepted clinical criteria. Length of survival was ascertained from death certificates or by contacting relatives. Ten-year survival after disease onset was 29% for both disorders. Median survival was 7.0 years (PSP) and 7.5 (MSA). Neither age, symptoms at onset, or disability at diagnosis predicted survival. At diagnosis, all PSP patients had oculomotor palsy, whereas 89% of MSA patients had dysautonomia; bradykinesia and falls were the most frequent common signs. Distinctive early signs were palilalia, cognitive impairment and hyperreflexia in PSP; hypophonia, anterocollis and dysautonomia in MSA. MSA patients responded better to levodopa. Attention to early distinctive features can improve differential diagnosis and inform subsequent management. PMID- 11731879 TI - Cutis verticis gyrata, mental retardation and Lennox-Gastaut syndrome: a case report. AB - Cutis verticis gyrata (CVG) is an abnormality of the scalp characterized by the formation of furrows and folds which cannot be flattened by traction or pressure. Primary and secondary forms of CVG have been described. We report on a patient affected by cutis verticis gyrata, mental regression and Lennox-Gastaut syndrome (LGS). Serum hormonal levels, karyotype and X fragile studies were normal. Magnetic resonance imaging of the brain showed only atrophic changes. The etiology of primary CVG remains unknown as does its relation with LGS. PMID- 11731880 TI - Extraskeletal Ewing's sarcoma presenting with neurological symptoms. Report of two cases. AB - Extraosseous Ewing's sarcoma (EES) is an uncommon condition that may develop in soft tissues at any location. Neurological symptoms leading to diagnosis have been reported exceptionally. We report two cases of EES in patients aged 16 and 74 years, presenting with cauda equina syndrome and radicular ulnar involvement respectively. Radiological images showed a paraspinal mass in both cases and biopsy disclosed the histological features of Ewing's sarcoma. EES should be kept in mind in the differential diagnosis of paraspinal soft tissues tumours at any age. PMID- 11731881 TI - Phenytoin hypersensitivity syndrome. AB - Phenytoin hypersensitivity syndrome (PHS) is a rare, and important entity characterized by rash, fever, lymphadenopathy, leukocytosis with atypical lymphocytes, eosinophilia and associated hepatitis. In this article, we present the clinical, laboratory and histopathologic results of 5 cases of PHS. In therapy, pheyntoin was stopped and sodium valproate (10-20 mg/kg day) was started. Additionally, prednisolone was given in two patients who had not resolved eruption with conservative therapy. PMID- 11731882 TI - Malignant peripheral nerve sheath tumor of the cauda equina. AB - Only one case of malignant peripheral nerve sheath tumor (MPNST) affecting the cauda equina region has been reported earlier. A 32-year-old male with congenital multiple subcutaneous swellings presented with low back pain, progressive paraparesis and bladder-bowel dysfunction. Magnetic resonance imaging (MRI) demonstrated a heterogeneously enhancing intradural lesion at L2-L4. At operation, on opening the dura, multiple nodular, firm matted masses attached to the lumbosacral nerve roots were encountered. Peripheral lesions were partially excised. Histopathological exam revealed varied cellularity with necrosis and pleomorphic nuclei suggestive of MPNST. MRI features, pathophysiological characteristics and the literature are reviewed. PMID- 11731883 TI - Reversible central pain. AB - We report two cases of central pain which receded completely after treatment of the inciting lesion. These cases highlight the intrinsic reversibility of central pain and the focal nature of central pain mechanisms. PMID- 11731884 TI - Migraine and arterial dissection in a young woman. AB - Ischemic stroke in young adults is rare (5%-10% of all ischemic strokes) and, in absence of other risk factors, may be associated with migraine. We describe the case of a 34-year-old woman, with a history of migraine without aura, who presented a sudden onset of headache with Horner's syndrome, and in whom neuroimaging showed evidence compatible with fibromuscular dysplasia (FMD) and arterial dissection of the extracranial internal carotid artery (ICA) and the carotid siphon. In our opinion, in young women with a long history of migraine, a careful study of the extracranial and intracranial arteries would be useful, although the cost/benefit ratio does not at present justify such a procedure. Our aim in the future is, therefore, to study a larger sample of migraine patients in order to find those patients who are most at risk of arterial dissection and who should, consequently, be carefully studied. PMID- 11731885 TI - Rivastagmine-induced agitation following transient recollection of autobiographical memory. PMID- 11731886 TI - Parenchymatous neurosyphilis. PMID- 11731887 TI - Incoercible topiramate-related vomiting in a patient with epilepsy and mental retardation. PMID- 11731888 TI - EXIT procedure in a twin gestation and review of the literature. AB - Prenatal diagnosis can show masses of the fetal neck, mouth, and face that can potentially cause respiratory distress at birth. To prevent such an emergency, the EXIT (ex utero intrapartum technique) is performed: it is the intrapartum intubation of the fetus at term while still connected to the placenta. The EXIT procedure was first performed in a case of cervical teratoma. Up to now a total of 34 cases are described, mostly cervical teratomas (13 cases), lymphangiomas (7), epignathus (3); babies' outcome has been successful in 25 of them, with one death related to the procedure. Among the reported cases we are aware of only one where EXIT was performed in a twin gestation, in which the normal twin was delivered first. In our case the normal fetus was posterior to the twin with cervical malformation, requiring us to work on the latter while the former was still in the uterus. After having safely secured the airway in twin A, twin B was prompt delivered with excellent general conditions. Our limited experience enlarges the possibility to perform this prenatal procedure even in "nonstandard" conditions, such as a twin gestation, and may prove useful to those who are going to deal with such issues. PMID- 11731889 TI - Perinatal management of fetal cardiac anomalies in a specialized obstetric pediatrics center. AB - Perinatal teams dealing with fetal heart disease frequently wonder which pregnancies might be terminated, and when delivery should take place in a specialized surrounding. We present a retrospective study of 229 fetuses, in which prenatal ultrasound showed a cardiac anomaly not compatible with a standard maternity ward delivery. One hundred nineteen pregnancies were terminated (group I) while 110 pregnancies led to the birth of a live baby (group II). Pathology in group I was discovered earlier than in group II (24 vs. 29.3 weeks' gestation; p <0.01), and associated malformations or chromosomal anomalies were much more frequent in group I (80/119 vs. 9/110; p <0.001). Among live born babies, three infants with transposition of the great arteries underwent Rashkind atrioseptostomy in the delivery room. With a minimum follow-up of 12 months, 69 children (63%) have undergone surgery. Among 92 survivors (1 child is lost to follow-up), 78 (71%) are asymptomatic and 14 symptomatic. Early prenatal diagnosis of fetal heart anomalies significantly facilitates prenatal work-up and perinatal care. We present the types of pathology having led to termination and define the situations in which children are at risk of perinatal hemodynamic compromise. PMID- 11731890 TI - First-trimester transvaginal ultrasonographic diagnosis of Dandy-Walker malformation. AB - We present transvaginal ultrasonographic findings of a fetus with Dandy-Walker malformation and associated massive obstructive hydrocephalus at 13 weeks' gestation. First-trimester ultrasonographic diagnosis of Dandy-Walker malformation is uncommon with only two such occurrences having been reported previously. These cases and recent reports of single gene transmission of this condition in some families emphasize the importance of first-trimester transvaginal ultrasound assessment especially in women with previously affected fetuses. PMID- 11731891 TI - Response to hepatitis B vaccine in preterm infants: four-dose schedule. AB - The objective of this study is to test a four-dose Hepatitis B vaccine schedule in premature infants (PI) and assess the immunogenicity of the vaccine with this schedule. We studied 29 PI who received the vaccine against Hepatitis B at birth, 1, 5, and 9 months of age. Antibodies against surface antigen (Anti-HBs) were measured before the third and fourth doses and 12 weeks after the fourth dose. Levels higher than 10 mIU/mL were considered protective, whereas more than 100 mIU/mL was an excellent response. Twenty-nine PI were studied. The average weight at the time of the initial dose was 1398 g, gestational age of 32.5 weeks, and a postnatal age of 9 days. Since the initial measurement, protective levels were achieved in all patients. The response was excellent in 24.1, 75.9, and 89.7%, after the second, third, and fourth doses, respectively. No correlation was found between the type of response and the infants' weight or the postnatal age. The Anti-HBs geometrical levels were 51.9, 133, and 133 mIU/mL after the second, third, and fourth doses, respectively. The favorable results obtained might be due to: the four-dose schedule itself, the dose we used (10 microg), the time interval between the doses, and even a race factor. Our schedule seems to be useful for PI infants and probably is no longer necessary to delay the vaccination, although this should be confirmed by further studies. PMID- 11731892 TI - Detection of interleukin-6 in maternal plasma predicts neonatal and infectious complications in preterm premature rupture of membranes. AB - The objective of this study is to determine if the detection of interleukin-6 (IL 6) in maternal plasma prior to delivery predicts neonatal and/or infectious complications in patients with preterm premature rupture of membranes. Patients with preterm premature rupture of membranes between 24 and 35 weeks' gestation were asked to participate in the study. Maternal blood was obtained prior to delivery. All patients received Ampicillin-sulbactam and steroids. IL-6 concentrations were determined by enzyme-linked immunoadsorbent assay (ELISA) using 50 mL of plasma assayed in duplicate. ELISA sensitivity was 18 pg/mL. Neonatal and infectious complications examined were respiratory distress syndrome, necrotizing enterocolitis, intraventricular hemorrhage, intra-amniotic infection, presumed neonatal sepsis, neonatal sepsis, and congenital pneumonia. Fifty-seven patients' plasma was analyzed. Thirty-five had positive plasma IL-6 prior to delivery. Twenty-seven patients had at least one neonatal complication with 24 (89%) being positive for IL-6. Of the 30 patients without complications, only 11 (37%) were positive (p = 0.0001, OR 13.8. 95% CI, 2.93-74.7). A subanalysis of patients who received a course of corticosteroids was performed and significance was maintained. Ten of 13 patients (77%) with neonatal complications had positive IL-6 compared with 40% without complications (p 3.0 microM). The most active compound was 4,6,4'-triacetyl-3',5'-dimethoxy-2-aurone with IC(50) values of 0.007 microM and 0.18 microM for the P. falciparum strains K1 and NF54, respectively. Interestingly, the multiple drug-resistant P. falciparum strain K1 was more sensitive to tested aurones than the drug susceptible strain NF54. PMID- 11731913 TI - Evaluation of in vitro activity of aurones and related compounds against Cryptosporidium parvum. AB - The efficacy of a series of aurones, auronols and 4-methoxy-alpha-pyrones has been screened for the ability to inhibit the intracellular growth of the parasitic protist Cryptosporidium parvum using an in vitro enzyme linked immunosorbent assay (ELISA). All aurones of this series were active at 25 to 100 microM. 10 of 19 aurones inhibited the intracellular growth of C. parvum by > 90 % with moderate to no toxicity. The most active of these was 3',4',6-trihydroxy-2 [phenylmethylene]-3(2H)-benzofuranone. PMID- 11731914 TI - A mechanistic approach to the in vivo anti-inflammatory activity of sesquiterpenoid compounds isolated from Inula viscosa. AB - The present study was designed to examine the anti-inflammatory activity of the sesquiterpenoids ilicic acid and inuviscolide, isolated from Inula viscosa, on cell degranulation, leukotriene biosynthesis, neurogenic drive and glucocorticoid like interactions. Swiss female mice were used to measure the ear oedema induced by phorbol esters or ethyl phenylpropiolate (EPP), and the paw oedema induced by phospholipase A(2) (PLA(2)) or serotonin. Drug treatment consisted of one topically-applied dose in the ear models and a subcutaneous or intraperitoneal injection in the paw models. Quantitative analysis of leukotriene B(4) (LTB(4)) formation was performed on rat peritoneal neutrophils by high performance liquid chromatography (HPLC). The lactone inuviscolide reduced the PLA(2)-induced oedema (ID(50): 98 micromol/kg). The effect on serotonin-induced oedema was not changed by modifiers of the glucocorticoid response. Ilicic acid showed minor in vivo effects, but was slightly more potent than inuviscolide on the 12-O tetradecanoylphorbol 13-acetate (TPA) acute oedema test (ID(50): 0.650 micromol per ear). Inuviscolide reduced LTB(4) generation in intact cells, with an IC(50) value of 94 microM. On the basis of the reported results, inuviscolide is the main anti-inflammatory sesquiterpenoid from Inula viscosa, and may act by interfering with leukotriene synthesis and PLA(2)-induced mastocyte release of inflammatory mediators. PMID- 11731915 TI - Isolation from Cussonia barteri of 1'-O-chlorogenoylchlorogenic acid and 1'-O chlorogenoylneochlorogenic acid, a new type of quinic acid esters. AB - 1'-O-Chlorogenoylchlorogenic acid and 1'-O-chlorogenoylneochlorogenic acid, a new type of quinic acid esters, have been isolated, in addition to six known quinic acid esters, rutin, and a mixture of saponins, from the methanol extract of Cussonia barteri Seemann (Araliaceae) leaves collected in Cameroon. Structure determination was achieved by NMR, mass, IR, and UV spectroscopy. All compounds were tested for inhibitory activity on 5-lipoxygenase and cyclooxygenase-1, for antimicrobial activity against Bacillus subtilis, Pseudomonas fluorescens, and Cladosporium cucumerinum, and for haemolytic activity. PMID- 11731916 TI - Isolation of three high molecular weight polysaccharide preparations with potent immunostimulatory activity from Spirulina platensis, aphanizomenon flos-aquae and Chlorella pyrenoidosa. AB - This research describes the identification of three new high molecular weight polysaccharide preparations isolated from food-grade microalgae that are potent activators of human monocytes/macrophages: "Immulina" from Spirulina platensis, "Immunon" from Aphanizomenon flos-aquae, and "Immurella" from Chlorella pyrenoidosa. These polysaccharides are structurally complex and have estimated molecular weights above ten million daltons. All three polysaccharides are highly water soluble and comprise between 0.5 % and 2.0 % of microalgal dry weight. Immunostimulatory activity was measured using a transcription factor-based bioassay for nuclear factor kappa B (NF-kappa B) activation in THP-1 human monocytes/macrophages. Using this system the EC(50) values for these microalgal polysaccharides are between 20 and 110 ng/ml (about 10pM). THP-1 activation was confirmed by measuring immune cytokine mRNA induction using reverse transcriptase polymerase chain reaction (RT-PCR). Each polysaccharide substantially increased mRNA levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF alpha). These polysaccharides are between one hundred and one thousand times more active for in vitro monocyte activation than polysaccharide preparations that are currently used clinically for cancer immunotherapy. PMID- 11731917 TI - Phenolic compounds with anti-inflammatory activity from Eupatorium buniifolium. AB - Anti-inflammatory activity was detected in the CH(2)Cl(2) extract of the aerial parts of Eupatorium buniifolium using the TPA-mouse ear model. Three compounds isolated from this extract, by bioassay-guided fractionation, significantly inhibited the inflammatory response. The compounds were identified as 5,7,5' trihydroxy-3,6,2',4'-tetramethoxyflavone (1), scopoletin (2) and centaureidin (3) which inhibited the edema by 67.3 %, 59.8 % and 49.7 %, respectively, at a dose of 1 mg/ear. PMID- 11731918 TI - In vitro anti-inflammatory effects of quercetin 3-O-methyl ether and other constituents from Rhamnus species. AB - The anti-inflammatory activities of the isolated flavonoids, quercetin 3-O-methyl ether (1), kaempferol (2), and quercetin (3), of Rhamnus nakaharai, and anthraquinone, frangulin B (4), of Rhamnus formosana, were assessed in vitro by determining their inhibitory effects on the chemical mediators released from mast cells, neutrophils, macrophages, and microglial cells. Compounds 1 - 3 strongly inhibited the release of beta-glucuronidase and lysozyme from rat neutrophils stimulated with formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB). Compound 1 strongly inhibited superoxide anion formation in fMLP/CB or phorbol 12-myristate 13 acetate (PMA)-stimulated rat neutrophils. Compound 1 exhibited potent inhibitory effect on tumor-necrosis factor-alpha ( TNF-alpha) formation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells while 1 and 4 showed potent inhibitory effects on TNF-alpha formation in LPS/IFN-gamma (interferon-gamma) stimulated murine microglial cell lines N9. PMID- 11731919 TI - Osteoblastic proliferation stimulating activity of Psoralea corylifolia extracts and two of its flavonoids. AB - Psoralea corylifolia L. fruit extracts exhibited osteoblastic proliferation stimulating activity in UMR106 cell line cultured in vitro. The flavonoids of corylin and bavachin were isolated and identified as active principles by activity-guided fractionation. The results suggested that Psoralea corylifolia L. fruit extracts and corylin and bavachin might stimulate bone formation or have potential activity against osteoporosis. PMID- 11731920 TI - Bakuchiol: a hepatoprotective compound of Psoralea corylifolia on tacrine-induced cytotoxicity in Hep G2 cells. AB - Bioassay-guided fractionation of the H(2)O extract of the seeds of Psoralea corylifolia furnished one hepatoprotective compound, bakuchiol (1), together with two moderately active compounds, bakuchicin (2) and psoralen (3), on tacrine induced cytotoxicity in human liver-derived Hep G2 cells. The EC(50) values of compounds 1 - 3 are 1.0, 47.0, 50.0 microg/ml, respectively. Silymarin as a positive control showed the EC(50) value with 5.0 microg/ml. PMID- 11731921 TI - Dammarane triterpene saponin from Bacopa monniera as the superoxide inhibitor in polymorphonuclear cells. AB - The hydroalcoholic extract of the whole plant of Bacopa monniera Wettst. (Scrophulariaceae), exhibited an inhibitory effect on superoxide released from polymorphonuclear (PMN) cells in the nitroblue tetrazolium (NBT) assay. The major saponin bacoside A(3) was found to be responsible for this effect in the herb. This compound showed 85, 91.66, 91.66, and 83 % inhibitions of NBT reduction at the concentrations of 200, 100, 50, and 25 microg/ml, respectively, with an IC(50) value of 10.22 microg/ml. These inhibitory effects were compared with those of the standard positive controls, quercetin and ascorbic acid with IC(50) of 111 and 14.16 microg/ml, respectively. Another major saponin bacopasaponin C was found to be much less potent as compared to bacoside A(3) whereas the remaining two mixtures of saponins were found to be inactive. PMID- 11731922 TI - Apoptosis induced by isoliquiritigenin in human gastric cancer MGC-803 cells. AB - Isoliquiritigenin, which is possibly a principal anti-tumor constituent of licorice, a traditional Chinese herb, was examined for apoptosis-inducing activity in human gastric cancer MGC-803 cells. Typical morphological and biochemical features of apoptosis including cell shrinkage, chromatin condensation, DNA ladder formation, and appearance of apoptotic peaks (subG(1)) were observed in MGC-803 cells with isoliquiritigenin treatment. Using Fluo-3 and Rh123 as fluorescent probes, respectively, it was found that the intracellular free calcium concentration increased and the mitochondrial transmembrane potential (Deltapsi(m)) decreased in a dose-dependent manner in apoptotic cells. These results suggest that isoliquiritigenin induced apoptosis of MGC-803 cells through calcium- and Deltapsi(m)-dependent pathways, indicating that it is potentially useful as a natural anti-cancer agent. PMID- 11731923 TI - Hypotensive effect of chemical constituents from Aloe barbadensis. AB - Hypotensive effects of aloeemodin, aloin A, elgonica dimer A and bisbenzopyran from Aloe barbadensis have been studied. Aloeemodin has emerged as a potent hypotensive agent in current pharmacological investigations and caused 26 %, 52 %, and 79 % falls in mean arterial blood pressure at the corresponding doses of 0.5, 1, and 3 mg/kg in rats. The paper also describes the absolute configuration of elgonica dimer A (1). PMID- 11731924 TI - Cardioactive diterpenoids from the roots of Salvia amplexicaulis. AB - Five diterpenoids, three steroids, four triterpenoids and one flavonoid were isolated from the roots of Salvia amplexicaulis Lam. (Lamiaceae). Structures of these compounds were elucidated by spectroscopic analysis. The crude extract and the pure compounds were tested for cardiovascular parameters using Wistar Albino rats. The crude extract, and 7-oxo-abieta-9,12,14-triene, ferruginol, stigmast-4 en-3-one showed a vasodepressor effect. PMID- 11731925 TI - Effect of trans-dehydrocrotonin, a 19-nor-clerodane diterpene from Croton cajucara on experimental hypertriglyceridaemia and hypercholesterolaemia induced by Triton WR 1339 (tyloxapol) in mice. AB - The effect of trans-dehydrocrotonin (t-DCTN) from Croton cajucara Benth. was investigated in mice on Triton WR 1339 (tyloxapol)-induced hypercholesterolaemia and hypertriglyceridaemia. Mice treated with single application of tyloxapol (400 mg/kg, i.p.) demonstrated significantly increased blood levels of total cholesterol and triglycerides at 24 h and 48 h after its injection, compared to normal controls. These increases were found to be markedly suppressed in animals treated orally with 25 and 50 mg/kg t-DCTN or 100 mg/kg gemfibrozil, an established antihypercholesterolaemic drug. These results suggest that t-DCTN may be a suitable candidate for combating pathologies associated with hyperlipaemia. PMID- 11731926 TI - Anti-thrombosis effect of paeoniflorin: evaluated in a photochemical reaction thrombosis model in vivo. AB - In a photochemical reaction microvessel thrombosis model, the anti-thrombotic effect of PT was evaluated and the results showed that PF could significantly prolong thromobosis time. The anti-thrombotic effect of PF may relate to the inhibition of arachidonic acid metabolism, the increase of t-PA activity, and the protective effect against free radical. PMID- 11731927 TI - In vitro evaluation of the antibacterial activity of beta-triketones admixed to Melaleuca oils. AB - The in vitro antibacterial properties of mixtures of Australian tea tree oil and niaouli oil after adding the beta-triketone complex isolated from manuka oil were tested. MIC and MBC values for four different bacteria were determined applying the broth dilution method. Both Melaleuca oil mixtures showed good antimicrobial effects against Staphylococcus aureus and Moraxella catarrhalis, exceeding the effectiveness of myrtol, which is well established in the treatment of acute and chronic bronchitis and sinusitis. The death kinetics of S. aureus were determined to draw subtle comparisons between the mixtures. The kill rate data indicated that both Melaleuca oil mixtures achieved a complete kill within 240 min. PMID- 11731928 TI - Essential oils from Piper cernuum and Piper regnellii: antimicrobial activities and analysis by GC/MS and 13C-NMR. AB - The essential oils of Piper cernuum and Piper regnellii leaves were analyzed by gas chromatography-mass spectrometry (GC-MS) and the results were compared to that obtained by means of a program designed to analyse (13)C-NMR data of complex mixtures. Bicyclogermacrene (21.88 %)/beta-caryophyllene (20.69 %) and myrcene (52.60 %)/linalool (15.89 %) were the major constituents in essential oil from leaves of P. cernuum and P. regnellii, respectively. Both essential oils presented growth inhibitory activities against Staphylococcus aureus and Candida albicans. PMID- 11731929 TI - Two new sesquiterpene lactones from Montanoa tomentosa ssp. microcephala. AB - Two new sesquiterpene lactones: 8alpha-(4'-acetoxymethacryloyloxy)-3alpha,9beta dihydroxy-1(10)E,4Z,11(13)-germacratrien-12,6alpha-olide (1) and 8alpha-(2'E)-(2' acetoxymethyl-2'-butenoyloxy)-3alpha,9beta-dihydroxy-1(10)E,4Z,11(13) germacratrien-12,6alphaolide (2), together with the known zoapatanolide A were isolated from the aerial parts of Montanoa tomentosa Cerv. in La Llave et Lex ssp. microcephala (Sch. Bip. In K. Koch) V.A. Funk (Asteraceae). The structures of all compounds were established on the basis of 1D, 2D NMR, and EIMS analysis. PMID- 11731930 TI - A new lignan glycoside from Eleutherococcus senticosus. AB - A new lignan glycoside, named eleutheroside E(2) (1), has been isolated from the roots of Eleutherococcus senticosus (known as "Siberian ginseng"), along with isomaltol 3-O-alpha-D-glucopyranoside (2), eleutherosides B, E and E(1), and thymidine. The structure of 1 was established by spectral interpretations as episyringaresinol 4"-O-beta-D-glucopyranoside. Compound 2 is described here for the first time as a naturally occurring compound. PMID- 11731931 TI - A reversed phase high performance liquid chromatography method for the analysis of boswellic acids in Boswellia serrata. AB - An HPLC method for the separation of boswellic acids, the active constituents in Boswellia serrata resin has been developed. The first accurate determination of 6 individual acids was possible in the resin as well as in multi-component preparations. By using an acidic mobile phase, raised temperature and a 4 microm Synergi MAX-RP 80 A column the acids could be detected at levels as low as 0.9 microg/ml. The study of market products revealed significant variations in the content of these pharmacologically active compounds in commercial samples. PMID- 11731932 TI - Application of sequence characterized amplified region (SCAR) analysis to authenticate Panax species and their adulterants. AB - A 420-bp RAPD fragment from Panax quinquefolius was converted to a sequence characterized amplified region (SCAR) marker. The main difference between the SCAR of P. quinquefolius and its homolog in P. ginseng is the presence of a 25 bp insertion in the latter. Primers derived from this sequence were successfully used to authenticate six Panax species and two common adulterants. PMID- 11731933 TI - A molecular marker that is specific to medicinal rhubarb based on chloroplast trnL/trnF sequences. AB - "Da-Huang" (Radix et Rhizoma Rhei, medicinal rhubarb), a famous and important Traditional Chinese Medicine, has often been confused with the adulterant species in the same genus, Rheum. Through sequencing the trnL (UAA)/trnF (GAA) regions of chloroplast DNA of thirteen species of Rheum (three medicinal rhubarb species and ten adulterant ones), a molecular marker of the medicinal species was found. A pair of PCR primers based on the sequences, was thus designed, which amplified a highly specific DNA fragment in medicinal rhubarb exclusively, and absent in the adulterants at all under an optimized PCR condition. PMID- 11731934 TI - The dual origin and Siberian affinities of Native American Y chromosomes. AB - The Y chromosomes of 549 individuals from Siberia and the Americas were analyzed for 12 biallelic markers, which defined 15 haplogroups. The addition of four microsatellite markers increased the number of haplotypes to 111. The major Native American founding lineage, haplogroup M3, accounted for 66% of male Y chromosomes and was defined by the biallelic markers M89, M9, M45, and M3. The founder haplotype also harbored the microsatellite alleles DYS19 (10 repeats), DYS388 (11 repeats), DYS390 (11 repeats), and DYS391 (10 repeats). In Siberia, the M3 haplogroup was confined to the Chukotka peninsula, adjacent to Alaska. The second major group of Native American Y chromosomes, haplogroup M45, accounted for about one-quarter of male lineages. M45 was subdivided by the biallelic marker M173 and by the four microsatellite loci alleles into two major subdivisions: M45a, which is found throughout the Americas, and M45b, which incorporates the M173 variant and is concentrated in North and Central America. In Siberia, M45a haplotypes, including the direct ancestor of haplogroup M3, are concentrated in Middle Siberia, whereas M45b haplotypes are found in the Lower Amur River and Sea of Okhotsk regions of eastern Siberia. Among the remaining 5% of Native American Y chromosomes is haplogroup RPS4Y-T, found in North America. In Siberia, this haplogroup, along with haplogroup M45b, is concentrated in the Lower Amur River/Sea of Okhotsk region. These data suggest that Native American male lineages were derived from two major Siberian migrations. The first migration originated in southern Middle Siberia with the founding haplotype M45a (10-11-11-10). In Beringia, this gave rise to the predominant Native American lineage, M3 (10-11-11-10), which crossed into the New World. A later migration came from the Lower Amur/Sea of Okhkotsk region, bringing haplogroup RPS4Y-T and subhaplogroup M45b, with its associated M173 variant. This migration event contributed to the modern genetic pool of the Na-Dene and Amerinds of North and Central America. PMID- 11731935 TI - A cascade of complex subtelomeric duplications during the evolution of the hominoid and Old World monkey genomes. AB - Subtelomeric duplications of an obscure tubulin "genic" segment located near the telomere of human chromosome 4q35 have occurred at different evolutionary time points within the last 25 million years of the catarrhine (i.e., hominoid and Old World monkey) evolution. The analyses of these segments reported here indicate an exceptional level of evolutionary instability. Substantial intra- and interspecific differences in copy number and distribution are observed among cercopithecoid (Old World monkey) and hominoid genomes. Characterization of the hominoid duplicated segments reveals a strong positional bias within pericentromeric and subtelomeric regions of the genome. On the basis of phylogenetic analysis from predicted proteins and comparisons of nucleotide substitution rates, we present evidence of a conserved b-tubulin gene among the duplications. Remarkably, the evolutionary conservation has occurred in a nonorthologous fashion, such that the functional copy has shifted its positional context between hominoids and cercopithecoids. We propose that, in a chimpanzee human common ancestor, one of the paralogous copies assumed the original function, whereas the ancestral copy acquired mutations and eventually became silenced. Our analysis emphasizes the dynamic nature of duplication-mediated genome evolution and the delicate balance between gene acquisition and silencing. PMID- 11731936 TI - Human-specific duplication and mosaic transcripts: the recent paralogous structure of chromosome 22. AB - In recent decades, comparative chromosomal banding, chromosome painting, and gene order studies have shown strong conservation of gross chromosome structure and gene order in mammals. However, findings from the human genome sequence suggest an unprecedented degree of recent (<35 million years ago) segmental duplication. This dynamism of segmental duplications has important implications in disease and evolution. Here we present a chromosome-wide view of the structure and evolution of the most highly homologous duplications (> or = 1 kb and > or = 90%) on chromosome 22. Overall, 10.8% (3.7/33.8 Mb) of chromosome 22 is duplicated, with an average sequence identity of 95.4%. To organize the duplications into tractable units, intron-exon structure and well-defined duplication boundaries were used to define 78 duplicated modules (minimally shared evolutionary segments) with 157 copies on chromosome 22. Analysis of these modules provides evidence for the creation or modification of 11 novel transcripts. Comparative FISH analyses of human, chimpanzee, gorilla, orangutan, and macaque reveal qualitative and quantitative differences in the distribution of these duplications--consistent with their recent origin. Several duplications appear to be human specific, including a approximately 400-kb duplication (99.4%-99.8% sequence identity) that transposed from chromosome 14 to the most proximal pericentromeric region of chromosome 22. Experimental and in silico data further support a pericentromeric gradient of duplications where the most recent duplications transpose adjacent to the centromere. Taken together, these data suggest that segmental duplications have been an ongoing process of primate genome evolution, contributing to recent gene innovation and the dynamic transformation of genome architecture within and among closely related species. PMID- 11731938 TI - Herpes simplex virus lymphadenitis: case report and review of the literature. AB - Localized or regional necrotizing lymphadenitis is an extremely uncommon manifestation of herpes simplex virus (HSV) infection. We report a case of necrotizing HSV lymphadenitis in a patient with both common variable immunodeficiency and natural killer cell deficiency and review the literature on this unusual complication of HSV infection. PMID- 11731937 TI - Nucleotide diversity and haplotype structure of the human angiotensinogen gene in two populations. AB - Variation in the angiotensinogen gene, AGT, has been associated with variation in plasma angiotensinogen levels. In addition, the T235M polymorphism in the AGT product is associated with an increased risk of essential hypertension in multiple populations, making AGT a good example of a quantitative-trait locus underlying susceptibility to a common disease. To better understand genetic variation in AGT, we sequenced a 14.4-kb genomic region spanning the entire AGT and identified 44 single-nucleotide polymorphisms (SNPs). Forty-two SNPs were observed both in 88 white and in 77 Japanese unselected subjects. Six major haplotypes accounted for most of the variation in this region, indicating less allelic complexity than in many other genomic regions. Although the two populations were found to share all of the major AGT haplotypes, there were substantial differences in haplotype frequencies. Pairwise linkage disequilibrium (LD), measured by the D', r(2), and d(2) statistics, demonstrated a general pattern of decline with increasing distance, but, as expected in a small genomic region, individual LD values were highly variable. LD between T235M and each of the other 39 SNPs was assessed in order to model the usefulness of LD to detect a disease-associated mutation. Among the Japanese subjects, 13 (33%) of the SNPs had r(2) values >0.1, whereas this statistic was substantially higher for the white subjects (occurring in 35/39 [90%]). LD between a hypertension-associated promoter mutation, A-6G, and 39 SNPs was also measured. Similar results were obtained, with 33% of the SNPs showing r(2)>0.1 in the Japanese subjects and 92% of the SNPs showing r(2)>0.1 in the white subjects. This difference, which occurs despite an overall similarity in LD patterns in the two populations, reflects a much higher frequency of the M235-associated haplotype in the white sample. These results have important implications for the usefulness of LD approaches in the mapping of genes underlying susceptibility to complex diseases. PMID- 11731939 TI - Defining opportunistic invasive fungal infections in immunocompromised patients with cancer and hematopoietic stem cell transplants: an international consensus. AB - During the past several decades, there has been a steady increase in the frequency of opportunistic invasive fungal infections (IFIs) in immunocompromised patients. However, there is substantial controversy concerning optimal diagnostic criteria for these IFIs. Therefore, members of the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group formed a consensus committee to develop standard definitions for IFIs for clinical research. On the basis of a review of literature and an international consensus, a set of research-oriented definitions for the IFIs most often seen and studied in immunocompromised patients with cancer is proposed. Three levels of probability are proposed: "proven," "probable," and "possible." The definitions are intended for use in the context of clinical and/or epidemiological research, not for clinical decision making. PMID- 11731940 TI - Geographic, demographic, and seasonal differences in penicillin-resistant Streptococcus pneumoniae in Baltimore. AB - We examined the epidemiology of invasive penicillin-resistant Streptococcus pneumoniae (PRSP) infections among residents of the Baltimore metropolitan area from 1995 through 1997. During this period, the proportion PRSP cases increased 42%, from 5.7% to 8.1% of cases. PRSP rates were highest among persons aged <5 and > or =65 years, black patients, and urban dwellers. However, the proportion of PRSP cases was higher among white persons (10%) than it was among black persons (5%) and among residents of suburban counties (10%) versus urban counties (6%). PRSP cases were more common in November-April (8%) than they were in May October (5%), particularly for persons aged > or =65 years (10% vs. 1%). By use of logistic regression, white race, suburban residence, and winter respiratory season were found to be independent predictors of infection with PRSP. The incidence of PRSP is increasing in Baltimore, and the seasonality of PRSP suggests that recent antibiotic use, which is more common in winter months, may rapidly affect the prevalence of resistant pneumococcal infections. PMID- 11731941 TI - The serological response of patients infected with the agent of human granulocytic ehrlichiosis. AB - To characterize the serological response in humans to human granulocytic ehrlichiosis (HGE), we prospectively observed 152 patients for as long as 42 months. HGE was confirmed by detection of morulae in blood smears, polymerase chain reaction, blood culture, or a combination of these tests for 94 patients (62.3%), and 92 (97.8%) of the patients had specific serum antibodies thereafter. One hundred twenty-six (99.2%) of 127 patients tested at 1 month were seropositive (89 of 127 patients had seroconversion), and 150 (98.7%) of the 152 patients had become seropositive by 6 months. Eleven patients (7.3%) remained seropositive at 42 months. Neither antibiotic therapy initiated during the first week of illness nor preexisting immunosuppressive conditions abrogated a serological response. Indirect fluorescent antibody testing of acute-phase and convalescent-phase serum samples is a sensitive tool for laboratory confirmation of HGE. PMID- 11731942 TI - Extension of the Lancefield classification for group A streptococci by addition of 22 new M protein gene sequence types from clinical isolates: emm103 to emm124. AB - Classic M protein serotyping has been invaluable during the past 60 years for the determination of relationships between different group A streptococci (GAS) strains and the varied clinical manifestations inflicted by these organisms worldwide. Nonetheless, during the past 20 years, the difficulties of continued expansion of the serology-based Lancefield classification scheme for GAS have become increasingly apparent. By use of a less demanding sequence-based methodology that closely adheres to previously established strain criteria while being predictive of known M protein serotypes, we recently added types emm94 emm102 to the Lancefield scheme. Continued expansion by the addition of types emm103 to emm124 are now proposed. As with types emm94-emm102, each of these new emm types was represented by multiple independent isolates recovered from serious disease manifestations, each was M protein nontypeable with all typing sera stocks available to international GAS reference laboratories, and each demonstrated antiphagocytic properties in vitro by multiplying in normal human blood. PMID- 11731943 TI - Molecular detection of rifampin and ofloxacin resistance for patients who experience relapse of multibacillary leprosy. AB - Molecular detection of rifampin resistance (rpoB analysis) in Mycobacterium leprae was determined for 49 patients who experienced relapse of multibacillary leprosy and for 34 untreated patients. Molecular detection of ofloxacin resistance (gyrA analysis) was determined for the 12 patients who experienced relapse and who had received ofloxacin. Results of molecular tests were compared with the reference susceptibility test in the mouse footpad. Overall, the efficiency of molecular detection--that is, positive DNA amplification--was 95%, whereas that of the in vivo test was 55% (P<.001). Results of molecular detection and in vivo test were fully concordant when both were available--that is, for 35 rifampin--sensitive cases of leprosy (no rpoB mutation), 4 ofloxacin-sensitive cases (no gyrA mutation), 11 rifampin-resistant cases (rpoB missense mutations), and 1 ofloxacin-resistant case (gyrA mutation). rpoB and gyrA analysis appears to be an effective method for detection of rifampin and ofloxacin resistance in patients with leprosy. PMID- 11731944 TI - The value of suction drainage fluid culture during aseptic and septic orthopedic surgery: a prospective study of 901 patients. AB - There are no guidelines on the value of suction drainage fluid culture (SDC), and it is difficult to determine whether the organisms cultured from suction drainage fluid samples are pathogenic or simply contaminants. We performed 2989 cultures of suction drainage fluid samples obtained, during a 1-year period, from 901 patients who underwent aseptic or septic orthopedic surgery (946 operations). The culture results were analyzed to evaluate their ability to detect postoperative infection after aseptic operations or to detect either a persistent or new episode of sepsis in patients known to have infection. For aseptic operations, the sensitivity of SDC was 25%, the specificity was 99%, the positive predictive value was 25%, and the negative predictive value was 99%. For septic operations, the sensitivity of SDC was 81%, the specificity was 96%, the positive predictive value was 87%, and the negative predictive value was 94%. We conclude that, for aseptic orthopedic surgery, SDC is not useful in detecting postoperative infection. However, for septic orthopedic surgery, it is of clinical importance. PMID- 11731945 TI - Higher occurrence of hepatotoxicity and rash in patients treated with oxacillin, compared with those treated with nafcillin and other commonly used antimicrobials. AB - This study compared adverse drug reactions (ADRs) to oxacillin with those to nafcillin and other antibiotics. We reviewed the medical records of 222 children receiving outpatient parenteral antimicrobial therapy (OPAT) from February 1995 through June 1999. The diagnosis, antibiotics used, ADRs, action taken, and patient demographics were recorded. The most common ADRs were neutropenia (9.8%), rash (8.5%), and hepatotoxicity (3.8%). ADRs occurred more frequently in the oxacillin group (58.5%) than in the nafcillin group (29.3%; P=.004), the clindamycin group (12.5%; P<.001) and the "other" antibiotics group (14.4%; P<.001). Hepatotoxicity and rash occurred more frequently in the oxacillin group (22% and 31.7%, respectively) than in the nafcillin group (0% [P<.001] and 10.3% [P=.008]), the clindamycin group (1.4% [P<.001] and 8.3% [P=.001]), and the other antibiotics group (1.4% [P<.001] and 1.4% [P<.001]). On the basis of this retrospective analysis, oxacillin use in children was associated with a higher incidence of hepatotoxicity and rash, compared with the use of nafcillin and other intravenous antimicrobials. PMID- 11731946 TI - Effect of changes in antibiotic prescribing on patient outcomes in a community setting: a natural experiment in Australia. AB - This study examined whether a significant change in antibiotic use caused by an Australian government directive targeted at amoxicillin with clavulanic acid (AC) was associated with changes in prescription share, health care costs, and patient outcomes. We used an integrated database of computerized general practice medical records, which included data regarding 34,242 patients and 318,234 recorded patient visits. There were 15,303 antibiotic prescriptions provided to 9921 patients during a 4-year period, with AC prescribed for 1453 (14.6%) of these patients. A total of 5125 patient outcomes were identified. There was a shift away from best-practice antibiotic prescribing, and a significant association was identified between the rate and cost of process-of-care and patient outcomes and the decrease in AC-prescription share. This policy initiative created unintended changes in prescribing behavior, increased costs to the government, and a trend toward poorer patient outcomes. Detailed analyses are required before instigating initiatives aimed at changing clinicians' prescribing behavior. PMID- 11731947 TI - Postarthroscopy surgical site infections: review of the literature. AB - The use of arthroscopy for both diagnosis and operative intervention has been increasing steadily since its introduction in the 1970s. It is generally associated with fewer complications and shorter times to mobilization than are open procedures. Overall reported rates of complications are low (0.1%-0.6% of procedures). This review focuses on infectious complications of arthroscopy, which are rare (0.01%-0.48% of procedures) but result in significant morbidity for the patient when they occur. The most commonly reported causative organisms are staphylococci. Several outbreaks have been reported related to breaks in infection control or to contaminated instruments. Suggested risk factors include use of intra-articular corticosteroids, prolonged tourniquet time, patient's age >50 years, failure to prepare the surgical site again before conversion to arthrotomy, procedure complexity, and a history of previous procedures. However, most reports use variable and unclear definitions of infection, which makes it difficult to draw firm conclusions. PMID- 11731948 TI - "Endgame" issues for the global polio eradication initiative. AB - The polio eradication initiative, created after the World Health Assembly resolved, in 1988, to eradicate poliomyelitis globally by 2000, has made remarkable progress. From 1988 through 2000, the number of countries where polio was endemic decreased from >125 to 20, and the estimated number of polio cases decreased from 350,000 to <3500, for a percentage decrease of >99%. Wild-type 2 poliovirus has not been detected worldwide since October 1999, despite improving surveillance. The major focus of the eradication effort is to complete the task of stopping wild-type poliovirus transmission. Given the rapid progress made toward this goal, planning for the posteradication era has begun in earnest (1) to minimize the risk of reintroduction of virus into the population from laboratory stocks or long-term carriers, and (2) to prevent vaccine-derived polioviruses from circulating and causing outbreaks. This report summarizes the current thinking about these "endgame" issues, as put forth by the World Health Organization's technical advisory body for the initiative, the Technical Consultative Group on the Global Eradication of Poliomyelitis. PMID- 11731949 TI - Countering the posteradication threat of smallpox and polio. AB - After eradication, there is a small but finite risk that smallpox and/or poliomyelitis viruses could accidentally escape from a laboratory or be released intentionally. The reintroduction of either virus into a highly susceptible population could develop into a serious catastrophe. To counter such an occurrence will require the use of vaccine, perhaps in substantial quantities. In the United States, new stocks of smallpox vaccine are being procured and arrangements are being made for a standby production facility. Similar provisions need to be considered for polio. To counter an epidemic of polio will require the use of the oral vaccine, which is presently the World Health Organization recommended vaccine of choice for countries throughout the developing world. In these countries, its continued use is advised because of its ability to induce intestinal immunity, its ability to spread to other susceptible household members and to protect them, its ease of administration, and its low cost. PMID- 11731950 TI - Reservoirs of human immunodeficiency virus type 1: the main obstacles to viral eradication. AB - Highly active antiretroviral therapy (HAART) has led to profound decreases in morbidity and mortality rates in human immunodeficiency virus type 1 (HIV-1) infected persons, at least in the developed world. Many infected persons have plasma levels of HIV-1 RNA that are less than the limits of detection of most clinical assays as a result of combination antiretroviral therapy. Nonetheless, HIV-1 has not been eradicated by HAART. This has been shown to be because of latent HIV-1 replication-competent provirus in resting CD4+ T lymphocytes, cryptic viral replication below the limits of detection of most clinical assays, and, possibly, the presence of viral sanctuary sites. An understanding of these reservoirs for HIV-1 in the setting of virally suppressive HAART will be critical for the development of new approaches to induce HIV-1 remissions and for the exploration of the possibility of viral eradication in the future. PMID- 11731951 TI - Meningococcal disease and travel. AB - Invasive meningococcal disease, in both endemic and epidemic forms, is the cause of significant morbidity and mortality worldwide. Despite all advances in therapy, the fatality rate of meningococcal meningitis remains unacceptably high, between 5% and 10%, and a similar proportion suffers long-term neurological sequalae. Prevention of this rapidly fatal disease is of paramount importance. The use of the available internationally licensed meningococcal vaccines would be indicated for individuals with medical conditions that increase the risk of the disease and for travelers to high-risk countries. In the last 2 years, there has been a shift in the epidemic pattern of meningococcal disease during the Hajj (pilgrimage) season, with predominance of Neisseria meningitidis serogroup W135. Recent changes have been made in the policy issued by the Saudi Ministry of Health (Riyadh, Saudi Arabia), which requires visitors from all over the world arriving for purposes of umra and Hajj to show evidence of vaccination against meningitis with the quadrivalent meningococcal vaccine. PMID- 11731952 TI - Ischemic cardiovascular disease in persons with human immunodeficiency virus infection. AB - Persons with human immunodeficiency virus (HIV) infection might be at risk for ischemic cardiovascular disease (CVD). We reviewed the records of 16 HIV-infected persons with proven CVD (8 cases of angina and 8 cases of myocardial infarctions). This represents 1.7% of HIV-infected persons seen at our institution from 1 April 1999 through 25 April 2000. In comparison with 32 HIV infected age- and sex-matched controls, case patients had more risk factors for CVD (median number of risk factors for CVD, 3 versus 1; P<.001), lower nadir CD4+ lymphocyte counts (median, 101 cells/mm3 versus 278 cells/mm3; P=.02), and a longer duration of prior exposure to nucleoside analogs (median, 190 weeks versus 130 weeks; P=.02). There was no difference in the duration of exposure to protease inhibitors. Ischemic CVD occurs in HIV-infected persons and appears to be most closely associated with traditional risk factors for coronary artery disease (for example, hypertension and hypercholesterolemia). Lower CD4+ lymphocyte counts and duration of HIV infection might also be risk factors or markers for the development of ischemic CVD. PMID- 11731953 TI - Focal neurological disease in patients with acquired immunodeficiency syndrome. AB - Focal neurological disease in patients with acquired immunodeficiency syndrome may be caused by various opportunistic pathogens and malignancies, including Toxoplasma gondii, progressive multifocal leukoencephalopathy (PML), cytomegalovirus (CMV), and Epstein-Barr virus-related primary central nervous system (CNS) lymphoma. Diagnosis may be difficult, because the findings of lumbar puncture, computed tomography (CT), and magnetic resonance imaging are relatively nonspecific. Newer techniques have led to improved diagnostic accuracy of these conditions. Polymerase chain reaction (PCR) of cerebrospinal fluid specimens is useful for diagnosis of PML, CNS lymphoma, and CMV encephalitis. Recent studies have indicated the diagnostic utility of new neuroimaging techniques, such as single-photon emission CT and positron emission tomography. The combination of PCR and neuroimaging techniques may obviate the need for brain biopsy in selected cases. However, stereotactic brain biopsy, which is associated with relatively low morbidity rates, remains the reference standard for diagnosis. Highly active antiretroviral therapy has improved the prognosis of several focal CNS processes, most notably toxoplasmosis, PML, and CMV encephalitis. PMID- 11731954 TI - Detection of a drug-resistant human immunodeficiency virus variant in a newly infected heterosexual couple. AB - Although new infections with drug-resistant human immunodeficiency virus type 1 have been recently reported to occur consistently, this is the first documented case of secondary transmission of a drug-resistant variant from 1 untreated subject to his sexual partner. PMID- 11731955 TI - Chagasic meningoencephalitis in a patient with acquired immunodeficiency syndrome: diagnosis, follow-up, and genetic characterization of Trypanosoma cruzi. AB - Early diagnosis of the clinical reactivation of Chagas' disease in human immunodeficiency virus- and Trypanosoma cruzi-coinfected persons is fundamental for a good prognosis. Polymerase chain reaction rapidly and efficiently demonstrated the presence and elimination of T. cruzi in the cerebrospinal fluid of a patient with chagasic meningoencephalitis. Characterization of T. cruzi, directly and indirectly in blood and cerebrospinal fluid samples, demonstrated homogeneity of kinetoplast DNA and the presence of lineage 1 (T. cruzi II) in both parasite populations. PMID- 11731956 TI - Salmonella typhiin the past decade: learning to live with resistance. PMID- 11731958 TI - Schizophrenia and serological methods for diagnosis of toxoplasmosis. PMID- 11731959 TI - Rendering beef safe. PMID- 11731960 TI - Infectious disease pathology and the autopsy. PMID- 11731961 TI - Rotavirus cerebellitis? PMID- 11731962 TI - Kawasaki-like syndrome: abacavir hypersensitivity? PMID- 11731965 TI - Diagnosis of Streptococcus pneumoniae lower respiratory infection in hospitalized children by culture, polymerase chain reaction, serological testing, and urinary antigen detection. AB - A prospective study of 154 consecutive high-risk hospitalized children with lower respiratory infections was conducted to determine the clinical utility of a pneumolysin-based polymerase chain reaction (PCR) assay compared with blood and pleural fluid cultures and serological and urinary antigen tests to determine the incidence of Streptococcus pneumoniae. Whole blood, buffy coat, or plasma samples from 67 children (44%) tested positive by PCR. Sensitivity was 100% among 11 promptly tested culture-confirmed children and specificity was 95% among control subjects. Age, prior oral antibiotic therapy, and pneumococcal nasopharyngeal colonization did not influence PCR results, whereas several surrogates of disease severity were associated with positive tests. Although serological and urinary antigen tests had comparable sensitivity, specificity varied among infected children, and statistical agreement among all assays was limited. These findings support the use of PCR tests to evaluate the protective efficacy of pneumococcal conjugate vaccines and to identify promptly children with pretreated or nonbacteremic pneumococcal lower respiratory infections. PMID- 11731966 TI - Immediate hypersensitivity reactions after use of tuberculin skin testing. AB - This report describes the frequency and characteristics of serious or life threatening hypersensitivity reactions to the tuberculin skin test over an 11 year period through November 2000. There were 24 reports and no deaths, indicating that such reactions are rare (0.08 reported reactions per million doses of tuberculin). PMID- 11731967 TI - Esophageal ulceration due to cytomegalovirus infection in a patient with acute retroviral syndrome. AB - Opportunistic infections during primary infection with human immunodeficiency virus type 1 have occasionally been reported in the medical literature, and those caused by cytomegalovirus have tended to be severe and prolonged. We describe a 40-year-old man who had acute retroviral syndrome complicated by a severe cytomegalovirus-induced esophageal ulceration, which was successfully treated with total parenteral nutrition and ganciclovir in addition to highly active antiretroviral therapy. PMID- 11731968 TI - Mycoplasma pneumoniae pericarditis demonstrated by polymerase chain reaction and electron microscopy. AB - We report a case of pericarditis occurring in a 17-year-old girl who received a bone marrow transplant. The presence of Mycoplasma pneumoniae in the pericardial fluid was demonstrated by polymerase chain reaction, and Mycoplasma-like organisms were detected by electron microscopy. Because no other infectious agent could be found in the pericardial fluid, M. pneumoniae was presumed to be the etiologic agent of the pericarditis. PMID- 11731969 TI - African sleeping sickness in tourists returning from Tanzania: the first 2 Italian cases from a small outbreak among European travelers. AB - A recent cluster of cases of African trypanosomiasis in humans (HAT) has been reported in tourists (most of whom were European) returning from Tanzania; we describe the first 2 patients (both of whom were Italian travelers) with HAT, who have been treated successfully. Because neither vaccine nor drug prophylaxis is currently recommended and/or available for persons traveling to areas of endemicity, physicians should be alerted about this uncommon but potentially life threatening disease. PMID- 11731970 TI - The effect of thermal stress on the mating behavior of three Drosophila species. AB - Selection may act on the weakest link in fitness to change how a species adapts to an environmental stress. For many species, this limitation may be reproduction. After adult Drosophila melanogaster, Drosophila simulans, and Drosophila mojavensis males were exposed to varying levels of thermal stress well below those that endanger life, courtship and mating frequency declined. The regression coefficients of both courtship and mating success did not differ significantly between D. melanogaster and D. simulans males. In contrast, significant differences were present between the two cosmopolitan species and D. mojavensis. Courtship frequency decreased at a much slower rate in D. mojavensis than in D. melanogaster and D. simulans, and while heat-stressed D. mojavensis males continued to court, many did not mate. In the cosmopolitan species, courting males almost always mated successfully. Courtship behaviors, including wing waving, were observed in D. mojavensis at temperatures that prohibited flight, while flight, courtship, and mating were knocked out simultaneously in D. melanogaster. One possible explanation for decreased flight ability and courtship success may be the reduced heat shock response in the flight muscle tissue because Hsp70 expression was lowest in the thoracic tissue of both D. melanogaster and D. mojavensis. PMID- 11731971 TI - Torpor in free-ranging tawny frogmouths (Podargus strigoides). AB - Several small caprimulgiform birds (<80 g) are known to enter torpor, apparently to cope with a fluctuating supply of insect prey. Since the large Australian tawny frogmouth (Podargus strigoides; 381-556 g) is also insectivorous, we investigated its thermoregulatory behaviour and thermal biology to determine whether this species is also heterothermic. In an open woodland at approximately 1,000 m altitude, we equipped eight free-ranging birds with external temperature sensitive radio transmitters attached to an elastic harness to measure skin temperature (T(skin)). Core body temperature (T(b)) was measured in three of these birds fitted with an additional intraperitoneal transmitter. T(skin) was closely correlated with T(b), although T(skin) was usually several degrees below T(b). During the three coldest months of the year (June-August), shallow torpor with T(b) as low as 29.1 degrees C occurred frequently, whereas during spring and summer, torpor was not recorded. Torpor occurred either during the night and/or during the first half of the day. Night torpor bouts were initiated after a short activity period around dusk and lasted on average for about 7 h. Torpid birds always aroused before sunrise to either commence a second short foraging period or to fly directly to a day roost tree. After birds roosted, T(b) fell again around sunrise, and birds occasionally entered a second dawn torpor bout; however, in most cases, T(b) increased rapidly not long after entry, most likely due to passive heating by the sun. We conclude that despite their large body size and energetically conservative hunting strategy, tawny frogmouths, like several related caprimulgiform species, frequently enter shallow torpor when low T(a) demands high energetic costs for normothermic thermoregulation and likely reduces insect availability. PMID- 11731972 TI - On the mound of Macrotermes michaelseni as an organ of respiratory gas exchange. AB - Patterns and rates of air movements in the mounds and nests of Macrotermes michaelseni were studied using tracer methods. Wind is a significant source of energy for powering nest ventilation, despite the mound being a completely enclosed structure. Nests are ventilated by a tidal movement of air driven by temporal variation in wind speed and wind direction. Density gradients sufficiently steep to drive bulk flow by natural convection will be rare. However, metabolism-induced buoyant forces may interact with wind energy in a way that promotes homeostasis of the mound atmosphere. PMID- 11731973 TI - Comparison of methods for evaluating energy expenditure of incubating wandering albatrosses. AB - Measurements of incubation energetics can vary depending on the method used to measure metabolism of an incubating bird. Therefore, we evaluated the energy expenditure of six male and four female wandering albatrosses (Diomedea exulans Linnaeus) using doubly labeled water (DLW), the rate of mass loss, and estimates of metabolic water production derived from water influx rate (WIR). Incubation metabolic rates (IMR) determined with DLW (169+/-21 kJ x kg(-1) x d(-1) SD) were significantly lower than estimates derived from mass loss (277+/-46 kJ x kg(-1) x d(-1) SD) and WIR (males=289+/-60 kJ x kg(-1) x d(-1) vs. females=400+/-69 kJ x kg(-1) x d(-1) SD). Estimates of IMR from mass loss and WIR were similar to IMR (305+/-39 kJ x kg(-1) x d(-1) SD) determined by respirometry in a previous study, and IMR from DLW was similar to estimates based on heart rate (HR; 147+/-26 kJ x kg(-1) x d(-1) SD) determined in another study. Applying the different measurements of IMR to construct an energy budget, we estimate that a breeding pair of wandering albatrosses spends 124-234 MJ to incubate the egg for 78 d. Finally, IMRs determined with DLW and HR were similar to estimated basal metabolic rates derived from six different allometric equations, suggesting that heat production from adult maintenance metabolism is sufficient to incubate the egg. PMID- 11731974 TI - The energetics of endotrophic development in the frog Geocrinia vitellina (Anura: Myobatrachinae). AB - The energetics of endotrophic development, where the nutrition required to complete metamorphosis is provided solely by yolk, has seldom been quantified. The energy cost of development to metamorphosis of the endotrophic Australian frog Geocrinia vitellina was measured using bomb calorimetry and closed-system respirometry. Dry yolk had an energy density of 26.4 J x mg(-1), and an average 2.8-mm-diameter ovum contained 144 J. Incubation at 15 degrees C produced a froglet of 5.8 mm snout-vent length, containing 88 J in 87 d, with 11% of residual yolk in the gut, which is markedly less than the 50% recorded in another endotroph, Eleutherodactylus coqui. Geocrinia vitellina lost 56 J of metabolic energy during development to metamorphosis at 15 degrees C, and the total production efficiency was 61.0%. A review of published egg energy densities found a mean for amphibians of 25.1 kJ x g(-1), significantly lower than the mean of 27.1 kJ x g(-1) for reptiles. Moreover, available amphibian data suggest that endotrophic species have high yolk energy densities and low mass-specific rates of oxygen consumption relative to exotrophic species (with feeding larvae); consequently, large ovum size may not necessarily be prerequisite for endotrophic development. PMID- 11731975 TI - The influence of carotenoid acquisition and utilization on the maintenance of species-typical plumage pigmentation in male American goldfinches (Carduelis tristis) and northern cardinals (Cardinalis cardinalis). AB - Birds display a tremendous variety of carotenoid-based colors in their plumage, but the mechanisms underlying interspecific variability in carotenoid pigmentation remain poorly understood. Because vertebrates cannot synthesize carotenoids de novo, access to pigments in the diet is one proximate factor that may shape species differences in carotenoid-based plumage coloration. However, some birds metabolize ingested carotenoids and deposit pigments that differ in color from their dietary precursors, indicating that metabolic capabilities may also contribute to the diversity of plumage colors we see in nature. In this study, we investigated how the acquisition and utilization of carotenoids influence the maintenance of species-typical plumage pigmentation in male American goldfinches (Carduelis tristis) and northern cardinals (Cardinalis cardinalis). We supplemented the diet of captive goldfinches with red carotenoids to determine whether males, which are typically yellow in color, were capable of growing red plumage. We also deprived cardinals of red dietary pigments to determine whether they could manufacture red carotenoids from yellow precursors to grow species-typical red plumage. We found that American goldfinches were able to deposit novel pigments in their plumage and develop a striking orange appearance. Thus, dietary access to pigments plays a role in determining the degree to which goldfinches express carotenoid-based plumage coloration. We also found that northern cardinals grew pale red feathers in the absence of red dietary pigments, indicating that their ability to metabolize yellow carotenoids in the diet contributes to the bright red plumage that they display. PMID- 11731976 TI - Changes in the composition of the urine of yellow-vented bulbuls (Pycnonotus xanthopygos): the effects of ambient temperature, nitrogen, and water intake. AB - Uricotely (uric acid >50% of urinary nitrogen) in birds was once considered ubiquitous. However, Anna's hummingbirds (Calypte anna) have been shown to be an exception to this rule; under conditions of low ambient temperature (T(a)) and on a nitrogen-free diet, they increased their water intake and often became ammonotelic (ammonia >50% of urinary nitrogen). Our aim was to identify the effects of nitrogen intake, water intake, and T(a) on the ammonia excretion of yellow-vented bulbuls (Pycnonotus xanthopygos). We chose this predominantly frugivorous species because many of the characteristics of nectarivores that were used to explain increased ammonia excretion by C. anna are also characteristics of frugivorous birds. We assayed ureteral urine composition in eight yellow vented bulbuls (P. xanthopygos), each randomly allocated a diet of 20% (0.6 M) sucrose solution supplemented by either 1.03 g/L or 7.23 g/L soy protein and held at a T(a) of either 28 degrees C or 10 degrees C. Food, and therefore water, intake rates varied with nitrogen intake but not with T(a). Food intake increased significantly with decreased nitrogen intake, while concentrations of all the excretory compounds in the urine (P<0.05) decreased; yet their proportions in the urine did not change significantly. The lower T(a) had no significant effect on food intake or on the concentration of uric acid. However, at 10 degrees C, the ammonia and urea concentrations increased (P<0.05), and this led to a significant increase in the proportion of ammonia in the urine. Our results demonstrate that, when bulbuls are exposed to low T(a), they are able to save energy by increasing the proportion of the ammonia in their urine. PMID- 11731977 TI - Cortisol effects on aerobic and anaerobic metabolism, nitrogen excretion, and whole-body composition in juvenile rainbow trout. AB - The influence of chronic cortisol elevation on metabolism, body composition, and fuel use patterns was examined in juvenile rainbow trout (Oncorhynchus mykiss). Measurements were performed in a control group (day 0) and in two experimental groups at days 3, 10, and 30 after treatment with a cortisol implant or a sham implant. All fish were fed 1% daily ration. Measured plasma cortisol levels were highest at day 3 and returned close to normal values by day 30 in cortisol implanted fish. No plasma cortisol elevation was observed in the sham group. Growth was depressed in the cortisol-treated fish. Cortisol elevation resulted in increased plasma glucose concentrations during the entire experimental period, elevated CO2 production at day 3 and 30, and an elevated respiratory quotient (RQ) exceeding 1.0 on these days. Nitrogen excretion, estimated as the sum of ammonia-N plus urea-N excretion, and the nitrogen quotient exhibited small decreases at day 30. Total-N excretion, measured with a nitrogen oxidizer, was approximately twice the sum of ammonia-N plus urea-N excretion but exhibited a similar trend. Aerobic metabolism (routine O2 consumption) was higher on day 10 compared to sham-implanted fish, although not relative to day 0 control levels. Anaerobic metabolism increased substantially, as evidenced by pronounced plasma lactate elevations at days 3 and 10, a small increase in whole-body lactate on day 10, and the elevated RQ on days 3 and 30. Body composition exhibited an increase in total carbohydrate at days 3 and 10, mainly reflecting increased glycogen levels. Protein concentration was stable, indicating, in accord with the respirometry data, that protein usage did not fuel the increased metabolism or carbohydrate elevation. Redirection of nutrient uptake from food and/or mobilization of lipid stores (which decreased relative to the control group but not relative to shams) are suggested as possible energy sources for these actions of cortisol. PMID- 11731978 TI - Seasonal changes of thermoregulation in the mole vole Ellobius talpinus. AB - Seasonal changes of thermoregulation were studied in mole voles (Ellobius talpinus Pall.) from southern Siberia. Spontaneous fluctuations of body temperature were significantly higher in summer than in autumn. Standard deviations from average daily temperature of individuals were 0.95 (N=212) in summer and 0.57 (N=248) in autumn. Thermoregulatory response to cold exposure and to norepinephrine injection varied in different seasons of the year. In cold seasons, mole voles are able to maintain temperature homeostasis in a wide range of environmental conditions. This ability declined only in summer. Hence, the mole vole's adaptation to the continental climate is based on distinct seasonal variations of thermoregulation. In summer, thermoregulatory mechanisms are similar to those of desert eusocial fossorial species, but in spring and autumn, mole voles share the thermoregulatory strategies of boreal terrestrial rodents. PMID- 11731979 TI - Torpor characteristics and energy requirements of furless Siberian hamsters. AB - After approximately 10 wk of exposure to decreasing day lengths, Siberian hamsters (Phodopus sungorus) begin to display spontaneous torpor bouts several times each week. Torpor is associated with reduced daily energy expenditure and lower food consumption and ameliorates the thermoregulatory challenges of winter. We tested the extent to which the energy savings conferred by daily torpor depend on the presence of an insulative pelage. Female hamsters were housed in a winter day length (8L:16D) at 5 degrees C; daily food intake and torpor characteristics were recorded for 5 wk in shaved (furless) or normal hamsters. Torpor-bout incidence decreased by 62% in furless hamsters, but the duration of individual bouts and the minimum body temperature attained during torpor were unaffected by loss of pelage. Body temperature declined more rapidly during entry into torpor and increased more slowly during arousal from torpor in furless than in control hamsters. Energy savings per torpor bout, assessed by the amount of food consumed on days that included a torpor bout, was substantially greater in normal than in furless hamsters (16.0% vs. 3.3%); this difference likely reflects the increased cost of thermoregulation during torpor, as well as the increased caloric expenditure incurred by furless hamsters during arousal from torpor. An insulative pelage may be a prerequisite for the energetic benefits derived from heterothermy in this species. PMID- 11731980 TI - Maximum titers of vitellogenin and total hemolymph protein occur during the canalized phase of grasshopper egg production. AB - Many organisms exhibit developmental plasticity only in sensitive phases and cannot respond to environmental perturbations at other times. However, we know little about the physiological events that define plastic and canalized phases. During egg production in insects, vitellogenin (Vg) accumulates first in the hemolymph and then in the eggs. In addition, storage proteins may be important resources for egg production. Therefore, we tested hypotheses on the relationships of Vg and TP (total hemolymph protein minus Vg) titers to the transition from flexible to inflexible development during egg production. In lubber grasshoppers, approximately 70% of TP is contained in three proteins that range from 68 to 83 kDa. We maintained females on food treatments that produced defined plastic and canalized periods, collected hemolymph every approximately 4 d, and determined the ages at which oviposition and the maximum Vg and TP titers occurred. Both Vg(max) titer and especially TP(max) titer were predictors of the number of eggs produced. The time from eclosion to Vg(max) was significantly affected by diet, but the time from Vg(max) to oviposition was not. Similarly, the time from eclosion to TP(max) was significantly affected by diet, while the time from TP(max) to oviposition was not. Hence, Vg(max) and TP(max) are physiological landmarks that occur during the canalized phase of egg production. PMID- 11731981 TI - Embryo tolerance and maternal control of the marsupial environment in Armadillidium vulgare (Isopoda: Oniscidea). AB - Marsupial development in terrestrial isopods subjects embryos to potential physiological stresses, including desiccation, osmotic variation, and high ammonia concentrations. In this study, we investigated tolerance of osmotic extremes, total ammonia, and pH in developmental stages of Armadillidium vulgare cultured in vitro. Marsupial stages were classified as stage 1 (chorionated eggs), stage 2 (having shed the chorion), and stage 3 (mancas). All stages showed wide but differing tolerance ranges. Stage 1 eggs possess the greatest ammonia tolerance, with high 7-d survival in 150 mM total ammonia, and a wide pH tolerance range. Mancas show the widest osmotic tolerance (100-1,400 mosm x kg( 1)) and display proficient hemolymph osmoregulation over this range. Stage 2 eggs reveal the narrowest tolerance ranges for all three parameters but still qualify as eurytopic. Silver staining revealed two distinct ion-transporting tissues in the developmental stages: a median band on the vitelline membrane of stage 1 and stage 2 eggs, corresponding in location to the embryonic dorsal organ, and the posterior three pairs of pleopodal endopodites in mancas. Gravid females do not downregulate ammonia but show efficient regulation of marsupial fluid pH and downregulation of osmolality during dehydration, both of which will provide additional protection to the marsupial young. PMID- 11731982 TI - Plasma urea, creatinine, and urea: creatinine ratio in reindeer (Rangifer tarandus tarandus) and in Svalbard reindeer (Rangifer tarandus platyrhynchus) during defined feeding conditions and in the field. AB - Variation in plasma urea and creatinine concentration and plasma urea:creatinine ratio (U:C) were studied in semidomestic free-ranging reindeer (Rangifer tarandus tarandus) on the Norwegian mainland, in wild Svalbard reindeer (Rangifer tarandus platyrhynchus), and in captive reindeer maintained either on a lichen-based diet or a protein-rich concentrate to investigate whether these parameters could be used as indicators of the nutritional status of reindeer. In the mainland animals, plasma creatinine concentration was high in winter and early spring and decreased by two-thirds toward the summer. The overall range in mean plasma creatinine concentration (+/-SE) was from 90+/-1.26 to 280+/-2.88 micromol/L. Mean plasma urea concentration (+/-SE) varied from 2.46+/-0.10 in winter up to 17.44+/-0.29 mmol/L in summer and autumn. Month of sampling explained 65% and 90% of the variation in plasma urea and creatinine concentrations, respectively, indicating that seasonality in the diet had the greatest influence on these parameters. Reindeer given lichens as the only feed showed an increase in plasma creatinine and a decrease in plasma urea concentration. Food restriction caused a temporary elevation in urea level but had no significant effect on plasma creatinine concentration. The slight effect of energy intake on urea and creatinine levels was supported by the fact that severe undernutrition in the Svalbard reindeer population had only a small effect on plasma urea and creatinine levels. Protein-rich pellet feed increased plasma urea from around 3 mmol/L to above 10 mmol/L and reduced creatinine concentrations to less than 100 micromol/L, suggesting that the protein content of forage is an important determinant of these blood parameters. Mean U:C ratio (+/-SE) in plasma varied from 8.9+/-0.28 to 120.8+/-1.88. Ratios above 20 appeared when protein intake was low and energy intake was restricted or when protein intake was high. Low ratios occurred when protein intake was low but energy intake adequate. Plasma urea and creatinine concentrations and the U:C ratio showed complex dynamics that were affected by both season and the protein and feed intake. We conclude that they appear to be difficult to interpret as single measures of nutritional status of reindeer. PMID- 11731983 TI - Thermoregulation in juvenile red kangaroos (Macropus rufus) after pouch exit: higher metabolism and evaporative water requirements. AB - The population dynamics of red kangaroos (Macropus rufus) in the Australian arid zone is tightly linked with environmental factors, which partly operate via the survival of juvenile animals. A crucial stage is the young-at-foot (YAF) stage when kangaroos permanently exit the pouch. We have examined the thermal biology of YAF red kangaroos during ages from permanent pouch exit until weaning. Over a wide range of environmental temperatures (ambient temperature [T(a)] -5 degrees to 45 degrees C), YAF red kangaroos had a mass-specific metabolism that was generally twice that of adults, considerably higher than would be expected for an adult marsupial of their body size. The total energy requirements of YAF red kangaroos were 60%-70% of those of adult females, which were three times their size. Over the same range in T(a), YAF red kangaroos also had total evaporative water losses equal to those of adult females. At the highest T(a) (45 degrees C), differences were noted in patterns of dry heat loss (dry conductance) between YAF red kangaroos and adult females, which may partially explain the relatively high levels of evaporative cooling by YAF. By weaning age, young kangaroos showed little change in their basal energy and water requirements (at T(a) 25 degrees C) but did show reduced mass-specific costs in terms of energy and water use at extremes of T(a) (-5 degrees and 45 degrees C, respectively). In their arid environment, typified by unpredictable rainfall and extremes of T(a), young red kangaroos may need to remain close to water points, which, in turn, may restrict their ability to find the high-quality forage needed to meet their high energy demands. PMID- 11731984 TI - Transfer of Vitamins E and A from yolk to embryo during development of the king penguin (Aptenodytes patagonicus). AB - Since the yolk lipids of the king penguin (Aptenodytes patagonicus) are rich in n 3 fatty acids, which are potentially susceptible to peroxidative damage, the yolk contents and yolk-to-embryo transfer of antioxidants and lipid-soluble vitamins were investigated under conditions of natural incubation in the wild. The concentration of vitamin E in the unincubated egg was 155 microg/g wet yolk, of which 88% was alpha-tocopherol and the rest was gamma-tocopherol. Vitamin A (2.9 microg/g) was present in the yolk entirely as retinol; no retinyl esters were detected. Throughout the latter half of the incubation period, vitamins E and A were taken up from the yolk into the yolk sac membrane (YSM) and later accumulated in the liver, with vitamin A being transferred in advance of vitamin E. In the YSM, vitamin A was present almost entirely as retinyl ester, indicating that the free retinol of the yolk is rapidly esterified following uptake. Retinyl esters were also the predominant form in the liver. The retinyl esters of the liver and YSM displayed different fatty acid profiles. At hatching, the brain contained relatively little vitamin E (4.7 microg/g) compared to the much higher concentration in the liver (482.9 microg/g) at this stage. Ascorbic acid was not detected in the yolk but was present at a high concentration in the brain at day 27 (404.6 microg/g), decreasing to less than half this value by the time of hatching. This report is the first to delineate the yolk-to-embryo transfer of lipid-soluble vitamins for a free-living avian species. The yolk fatty acids of the king penguin provide an extreme example of potential oxidative susceptibility, forming a basis for comparative studies on embryonic antioxidant requirements among species of birds whose yolk lipids differ in their degree of unsaturation. PMID- 11731985 TI - Proximate causes of intraspecific variation in locomotor performance in the lizard Gallotia galloti. AB - To understand the evolution of biological traits, information on the degree and origins of intraspecific variation is essential. Because adaptation can take place only if the trait shows heritable variation, it is important to know whether (at least) part of the trait variation is genetically based. We describe intra- and interindividual variation in three performance measures (sprint speed, climbing, and clambering speed) in juvenile Gallotia galloti lizards from three populations and examine how genetic, environmental (incubation temperature), and ontogenetic (age, size) effects interact to cause performance variation. Moreover, we test whether the three performance traits are intercorrelated phenotypically and genetically. Sprint speed is highest in juveniles incubated at the lowest temperature (26 degrees C) irrespective of population. Climbing speed differs among populations, and the differences persist at least until the lizards are 30 wk old. This suggests that the three populations experience different selective pressures. Moreover, mass, snout-vent length, and hindlimb length seem to affect climbing performance differently in the three populations. The variation in sprinting and climbing ability appears to be genetically based. Moreover, the two performance traits are intercorrelated and thus will not evolve independently from each other. Clambering speed (i.e., capacity to climb up an inclined mesh) varies among individuals, but the origin of this variation remains obscure. PMID- 11731986 TI - [Psychosomatic dermatology - state of the art]. AB - Based on the current literature, the paper gives an overview of the state of knowledge in psychosomatic dermatology. The authors propose the following classification: (1.) Dermatologic problems as a consequence of psychic disorders (e.g. factitious disorder, trichotillomania, dysmorphophobia, delusions of parasitosis etc.); (2.) Multifactorial dermatologic disorders, which can be substantially influenced by psychological factors (e.g. atopic dermatitis, psoriasis, chronic urticaria etc.); (3.) Psychic disorders developing as a consequence of disfiguring or life-threatening dermatoses (e.g. adjustment disorder, depressive disorder, anxiety disorder); (4.) Comorbidity with other psychiatric disorders. Psychosomatic dermatology developed from an anecdotal initial stage to more systematic research with larger numbers of patients, employing control groups and psychometric instruments. During the last decade, research focused on psychophysiological and psychoneuroimmunological mechanisms which influence dermatologic disorders. This course of development and the current state of knowledge are presented for atopic dermatitis and psoriasis. PMID- 11731987 TI - [Coping with conflict as pathogenetic link between psychosocial adversities in childhood and psychic disorders in adulthood]. AB - OBJECTIVES: The present study explores the connection between neurotic or immature strategies for coping with conflict in the presence of specific psychosocial adversities during childhood and their cumulative effect. Here we test the hypothesis as to whether certain coping strategies can be viewed as a pathogenetic link between psychic illness in adulthood and adversities during childhood. METHODS: 407 consecutive patients referred to the outpatient unit of a psychosomatic university hospital were included in the study. The strategies for coping with conflict were assessed by means of two psychometric methods (FKBS, Hentschel et al., 1996; SBAK, Ehlers u. Peter, 1989), and the childhood adversities were assessed using the Mainz Structured Biographic Interview (Egle, 1993). RESULTS: Patients with sexual abuse and/or physical maltreatment during the first 14 years of life showed significantly higher values for immature coping mechanisms than patients without such traumatisation. Childhood adversities correlated negatively with mature coping strategies and positively with immature strategies. Significant differences also arise when patients with low risk are compared to those with a higher amount of childhood adversities. The latter group exhibits significantly higher scores for immature neurotic coping strategies, while the former exhibits mature ones. By means of a multivariate logistic regression analysis, 7 and 10% respectively of the variance of the predominant coping strategies within each group occurring in adulthood could be accounted for. CONCLUSIONS: The results suggest that in patients exposed to psychosocial stress during childhood an increased vulnerability for psychic disorders in adulthood is, in part, caused by the use of immature strategies for coping with conflict. PMID- 11731988 TI - [Relationship, resistance and insight as depicted in measures concerning form and content of speech - a textual analysis of the course of a female in-patient with eating disorder]. AB - OBJECTIVES: This multi-methodical single case study examines how relationship, resistance and insight, the important therapeutic factors, are conveyed in a therapeutic process. METHODS: For depicting interaction the textual parameters - direct speech, acknowledgement tokens, activity of speech and personal pronouns - of Formal Psycholinguistic Text Analysis (Overbeck et al., 1996) are applied. The activity of the unconscious theme is measured by the Interferenzindex (Argelander, 1984). The link of emotional experiences with words is depicted by the Computerized Referential Activity method (Mergenthaler and Bucci, 1999). The attainment of emotional insight in the Model of the Therapeutic Cycle is perceived via the Emotion-/Abstraction-Patterns (Mergenthaler, 1997). RESULTS: Three phases are discriminated accordingly: Resistance, characterised by defensive intellectualization, symbolic representation on the basis of the secure relationship and emotional insight and detachment. CONCLUSIONS: The validity of the results in this single case study is guaranteed by the multi-methodical strategy. PMID- 11731989 TI - [The effects of psychodynamic inpatient treatments on personality data of patients with anorexia nervosa and bulimia nervosa - results of the German multicenter study (TR-EAT)]. AB - OBJECTIVES: How are the effects of a psychodynamic inpatient treatment on personality data of patients with anorexia nervosa and bulimia nervosa? In how many patients can a successful change in the personality area be observed? METHODS: 732 patients were assessed at the beginning and the end of an inpatient treatment as well as 2 1/2 years after this treatment. Data were collected by means of the personality inventories "Freiburger Persoenlichkeitsinventar (FPI R)" and "Narzissmusinventar". A definition of success was constructed using the clinical significance concept. It is related to the scales life satisfaction, inhibition (FPI-R), powerless self and negative body self (Narzissmusinventar). RESULTS: The results in both questionnaires show the clinically well known psychopathology characteristics of eating disorder patients. During the inpatient treatment and also afterwards, there are improvements in the personality data, but not to the extent of the values obtained from a healthy control group. In the outcome measure, one third of the patients experienced remarkable improvements. The success in the personality area is positively correlated to the success in the eating disorder symptoms. This success could not be predicted by initial data or by therapy data. CONCLUSIONS: There are marked impairments of the patients in the personality data. Changes in this domain take time and are rather small. PMID- 11731990 TI - [Course of symptom severity and prediction of outcome in inpatient psychotherapy]. AB - OBJECTIVES: The study looks for typical curves of symptom development in inpatients and investigates the influence of initial remoralisation and terminal improvement on the global therapy outcome. METHODS: 71 psychotherapy inpatients (affective, anxiety, eating and personality disorders; average treatment duration 11.9 weeks) gave weekly self-reports on symptom severity (BSI, Derogatis, 1983) and had a complete outcome evaluation. Regression parameters of the symptom curves were taken for correlational and path analytic calculations. RESULTS: Initial symptom alleviation is able to predict better global outcome. However there is a relevant subgroup of patients which has a good outcome despite an initial deterioration. CONCLUSIONS: Initial remoralisation predicts better outcome but is not a necessary condition. The results partly call into question the group statistical "dose-response-curves". Further studies should test the influence of other process parameters (group cohesion, therapeutic alliance etc.) on the course of treatment. PMID- 11731991 TI - [Perspectives of psychoanalytic psychosomatics]. AB - The paper discusses a variety of perspectives of psychoanalytic psychosomatics in the past, the present and the future. An epigenetic model of scientific development is introduced and developmental strains in psychosomatic medicine are evaluated according to the claims of the bio-psycho-social model. In historical terms, the psychological dimension of psychoanalytic psychosomatics has been the first strain to be elaborated; it is being extended still. The biological, somatic and bodily dimension of psychosomatic medicine was the next to be explored; during the last decade, this strain has found increasing interest, especially neurobiological research. Though the social dimension has not been neglected, it will be the main task for psychoanalytic psychosomatics to consider in the future. Likewise, a mandatory future challenge will be a more intensive discussion of the epistemological basis of psychosomatic medicine and psychoanalytic psychosomatics. The historical development of psychosomatic medicine is highlighted by examples drawn mainly from the history of Heidelberg Psychosomatic University Clinic that has its 50th anniversary in 2000. PMID- 11731993 TI - Genes and cholestasis. PMID- 11731994 TI - Carcinoembryonic antigen-related cell adhesion molecule 1 is the 85-kilodalton pronase-resistant biliary glycoprotein in the cholesterol crystallization promoting low density protein-lipid complex. AB - A pronase resistant 85-kd glycoprotein in the Concanavalin A-binding fraction (CABF) of biliary glycoproteins has been reported to act as a promotor of cholesterol crystallization. De Bruijn et al. (Gastroenterology 1996;110:1936 1944) found this protein in a low-density protein-lipid complex (LDP) with potent cholesterol crystallization promoting activity. This study identifies and characterizes this protein. An LDP was prepared from CABF by discontinuous gradient ultracentrifugation. Proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), blotting and immunochemical staining with anti-carcinoembryonic antigen, CEA-related adhesion molecule 1 (CEACAM1) cross-reacting antibodies. Biliary concentrations of CEA cross-reacting proteins were determined in patients with and without gallstones. Two isoforms of CEACAM1 (85- and 115-kd bands), CEA and 2 CEA cross-reacting protein bands of 40 and 50 kd were found in human bile. All bands were also present in CABF, but only a subfraction of the 85-kd band found in the LDP was resistant to digestion with pronase. CEACAM1-85 exhibited potent cholesterol crystallization promoting activity in vitro and accounted for most of the activity in CABF. Total CEA cross reacting protein concentrations were the same in gallbladder biles from patients with cholesterol and pigment gallstones but only half of those in biles from nongallstone subjects. In conclusion, we have identified the protein component of the cholesterol crystallization promoting LDP to be CEACAM1-85. PMID- 11731995 TI - The geographical distribution of primary biliary cirrhosis in a well-defined cohort. AB - The incidence of primary biliary cirrhosis (PBC) varies widely between regions. However, little is known about variation within regions and the degree to which this may reflect environmental risk factors. The aim of this study was to describe the spatial distribution of cases of PBC in a defined region of Northeast England over a defined period, and to assess the magnitude of any departure from random spatial distribution. Seven hundred seventy patients with established PBC were identified in a previous comprehensive case finding study. A total of 3,044 control locations were randomly selected from postcode (zip code) data weighted for number of drop off points per postcode. Geographical analysis was performed by testing both for spatial variation in risk and local clustering by using previously described point process methods. Both tests used the same null hypothesis that risk of disease does not vary spatially and cases occur independently of each other. Statistically significant spatial variations in risk were found in the whole study region (P <.001) and in the major urban area within the region (P <.004). Risk was higher in the urban area of Tyneside than in the surrounding rural area. Within the rural area, spatial variation in risk was equivocal (P =.012), but there was significant (P =.001) clustering of cases (estimated average cluster effect approximately 10 excess cases within a 7-km radius). PBC occurred to a density of 10.7 cases/km(2) in the highest risk areas. In conclusion, PBC is unevenly distributed in Northeast England. This may reflect one or more environmental risk factors in its etiology. PMID- 11731996 TI - Impact of gastroenterology consultation on the outcomes of patients admitted to the hospital with decompensated cirrhosis. AB - Managed care has strongly discouraged generalists from referring patients to specialists in an effort to reduce the costs of health care. The aim of this study was to compare patient outcomes when generalists work together with gastroenterologists or alone in the management of patients admitted to the hospital with decompensated cirrhosis. Consecutive patients admitted to the hospital with decompensated cirrhosis over a 1-year period were identified. We compared the length of stay, cost of hospitalization, incidence of hospital readmission, and mortality for patients who did and those who did not have a gastroenterology (GI) consultation. A GI consultation was requested for 107 of the 197 patients (54.3%). Patients who had a GI consultation had a significantly shorter length of stay (5.6 +/- 3.5 vs. 10.1 +/- 5.8 days, P <.001) and a lower cost of hospitalization ($6,004 +/- $4,994 vs. $10,006 +/- $6,183, P <.001) than those patients who were managed by generalists alone. The 30-day incidence of readmission (13.3% vs. 27.8%, P =.01) and mortality (7.5% vs. 16.7%, P =.045) were significantly lower in the GI consultation group. During a median follow-up of 618 days (range, 2-970), patients who had a GI consultation during hospitalization had a significantly longer time to hospital readmission (P <.001) and improved survival (P =.02) compared with those who were managed by generalists alone. In conclusion, for patients admitted to the hospital with decompensated cirrhosis, individuals who were managed by generalists in conjunction with gastroenterologists had better outcomes than those who were managed by generalists alone. PMID- 11731997 TI - Propranolol for the prevention of first esophageal variceal hemorrhage: a lifetime commitment? AB - Although beta blockers have had significant impact in the treatment of portal hypertension, the question of how long they should be continued for prevention of variceal hemorrhage remains unknown. Prospective studies on beta blockers to prevent variceal hemorrhage lack long-term follow-up, and indefinite administration of beta blockers for primary prevention of variceal bleeding has become standard practice. The aim of this study was to determine the outcomes of patients in whom beta blocker therapy was discontinued. Patients completing a prospective, randomized, double-blind, placebo-controlled trial of propranolol for the primary prevention of variceal hemorrhage were tapered off of propranolol and placebo and followed prospectively for subsequent events. Of the 49 patients in the follow-up study (25 former propranolol, 24 former placebo), 9 experienced variceal hemorrhage (6 former propranolol, 3 former placebo). Following withdrawal of propranolol, the freedom from variceal bleeding was not significantly different between these 2 groups of patients, suggesting that the protective effect of propranolol against variceal hemorrhage, noted previously, was no longer present. Seventeen patients died (12 former propranolol, 5 former placebo) during the follow-up study. Cumulative survival was longer in the placebo group. These trends for EVH and survival were opposite to those observed in the original study population while patients were taking medication. When propranolol is withdrawn, the risk of variceal hemorrhage returns to what would be expected in an untreated population. Patients who discontinue beta blockers experience increased mortality compared with an untreated population. These observations support the current practice of indefinite prophylactic therapy. PMID- 11731998 TI - Hepatic and intestinal cytochrome P450 3A activity in cirrhosis: effects of transjugular intrahepatic portosystemic shunts. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is performed to treat some complications of cirrhosis. This study investigated the effects of cirrhosis and TIPS on intestinal and hepatic cytochrome P450 3A (CYP3A) activity. Nine volunteers were cirrhotic patients with TIPS, 9 were cirrhotic controls (matched for sex, age, etiology, and Child-Pugh class), and 9 were sex- and age-matched healthy volunteers. Simultaneous doses of midazolam were given intravenously (0.05 mg/kg) and orally (3 mg of [15N3]midazolam). Peripheral and portal venous blood samples were assayed for midazolam and [15N3]midazolam. The systemic clearance of midazolam was significantly greater (P <.05) in healthy volunteers (0.42 +/- 0.10 L x h(-1) x kg(-1)) compared with cirrhotic controls (0.20 +/- 0.05) and with cirrhotic patients with TIPS (0.21 +/- 0.09). Hepatic availability followed the same trend. The bioavailability of midazolam was significantly higher (P <.05) in cirrhotic patients with TIPS (0.76 +/- 0.20) compared with cirrhotic controls (0.27 +/- 0.14) and with healthy volunteers (0.30 +/- 0.10). The intestinal availability was significantly greater (P <.05) in cirrhotic patients with TIPS (0.83 +/- 0.17) compared with cirrhotic controls (0.32 +/- 0.16) and with healthy volunteers (0.42+/-0.15). As expected, hepatic CYP3A activity was reduced in cirrhosis. However, in cirrhotic patients with TIPS, there was a marked loss in first-pass metabolism of midazolam as a result of diminished intestinal CYP3A activity. PMID- 11731999 TI - Contrast-enhanced Doppler ultrasonography in the diagnosis of hepatocellular carcinoma and premalignant lesions in patients with cirrhosis. AB - Hepatocellular carcinogenesis in cirrhosis is a multistage process that includes large regenerative nodules, dysplastic nodules, and hepatocarcinoma. The aim of this study was to establish whether contrast-enhanced Doppler ultrasonography (US) is able to distinguish between early hepatocellular carcinoma (HCC) and small nonmalignant nodules in cirrhosis. Between January 1998 and December 1999, 500 cirrhotic patients with no previous history of HCC or evidence of hepatic focal lesions were enrolled and prospectively followed-up with US every 6 months until December 2000. Sixty-one patients developed focal lesions, 12 multifocal, and 49 monofocal. Biopsy of focal lesions, contrast-enhanced Doppler US, and spiral computed tomography (CT) were performed in 41 consecutive patients with small (<3 cm) monofocal lesions. Twenty nodules were diagnosed as HCC and 21 as nonmalignant (14 large regenerative nodules, 3 low-grade, and 4 high-grade dysplastic nodules) by liver biopsy. Intratumoral arterial blood flow was detected in 19 of 20 (95%) HCC and 6 of 21 (28%) nonmalignant nodules by contrast enhanced Doppler US (P<.0001). The mean peak resistance and pulsatility indices were 0.82 +/- 0.09 and 1.56 +/- 0.2 in HCC and 0.62 +/- 0.08 and 0.82 +/- 0.08 in dysplastic lesions (P =.002 and.0001), respectively. Spiral CT revealed arterial perfusion in 19 of 20 HCC and in 4 of 21 nonmalignant nodules (high-grade dysplastic nodules). Four of the apparently false-positive nodules at enhanced Doppler US were high-grade dysplastic nodules and 2 evolved to HCC during follow up. In conclusion, contrast-enhanced Doppler US is a noninvasive, very sensitive technique in differentiating malignant and premalignant lesions from nonmalignant focal lesions in the liver. PMID- 11732000 TI - Patterns of regional sympathetic nerve traffic in preascitic and ascitic cirrhosis. AB - Overactivity of the sympathetic nervous system and portal hypertension are key factors in the development of ascites in cirrhosis. The sympathoexcitation that characterizes the more advanced stages of liver diseases is less clearly defined in preascitic cirrhosis. We measured sympathetic nerve traffic to skeletal muscle (peroneal nerve) and to skin districts by microneurography in (1) 12 Child class A cirrhotic patients with clinically significant portal hypertension (portal pressure gradient > 10 mm Hg, 14.8 +/- 1.2 mm Hg, mean +/- SEM) but without actual or previous ascites, (2) 16 Child class C cirrhotic patients with tense ascites, and (3) 10 patients with mild congestive heart failure, a condition paradigmatic of a marked sympathetic activation. Muscle sympathetic nerve traffic was markedly increased in Child class C subjects as compared with controls (23.9 +/- 1.6 bursts/min, P <.01) and superimposable to that recorded in heart failure patients (52.9 +/- 4.7 vs. 60.3 +/- 2 bursts/min, P = not significant). Muscle sympathetic nerve traffic was also increased in Child class A subjects (41.6 +/- 2 bursts/min, P <.01 vs. controls) although to a lesser extent (P <.05 vs. Child class C patients). Skin sympathetic nerve traffic was within the normal range in all patients. Neurohormones were all markedly increased in Child class C subjects. Only norepinephrine was increased in Child class A patients. Our data show that sympathetic nerve traffic activation (1) is already detectable in Child class A cirrhosis when clinically significant portal hypertension is present but ascites never developed and (2) is not generalized because although muscle traffic is increased, skin traffic is within normal range. The role of drugs modulating sympathoactivation should be investigated in preascitic cirrhosis. PMID- 11732001 TI - Activation of caspase-8 during N-(4-hydroxyphenyl)retinamide-induced apoptosis in Fas-defective hepatoma cells. AB - We observed that N-(4-hydroxyphenyl)retinamide (4HPR), a chemopreventive and chemotherapeutic agent, effectively induced apoptosis in hepatoma cells. Interestingly, Fas-negative (Hep 3B and PLC/PRF/5) hepatoma cells were shown to be more susceptible to apoptosis induced by 4HPR than were Fas-positive (Hep G2 and SK-HEP-1) hepatoma cells. Thus, we explored the mechanisms underlying 4HPR induced apoptosis in Fas-defective hepatoma cells. Hep 3B cells stably expressing the dominant-negative Fas-associated death domain (dnFADD) showed no alteration in 4HPR drug susceptibility, but when stably expressing E1B19K, Crm A, or dominant-negative FLICE (dnFLICE), Hep 3B cells were resistant, suggesting that 4HPR-induced apoptosis was mediated by caspase-8 activation. Furthermore, apoptosis could be completely blocked by Z-VAD-FMK (a general caspase inhibitor) or by IETD-CHO (a caspase-8 inhibitor), but was only partially blocked by Ac-DEVD CMK (a caspase-3 inhibitor), by N-acetyl-L-cysteine (NAC) (an antioxidant), by N acetyl-leucyl-leucyl-norleucinal (ALLN) (a calpain inhibitor I), or by Z-LEHD-FMK (a caspase-9 inhibitor). Time-sequence analysis of the induction of apoptosis by 4HPR revealed that an initial caspase-8 activation was followed by late mitochondrial cytochrome c release and minor caspase-9 activation, which suggested that caspase-8 activation is the primary upstream regulatory point. Activation of Bid or induction of proapoptotic Bax was not observed during apoptosis. In contrast, Bcl-xL expression was decreased during 4HPR-induced apoptosis. Taken together, these results indicate that 4HPR may be a potential chemotherapeutic drug, which is able to induce apoptosis in Fas-defective hepatoma cells through caspase-8 activation. PMID- 11732002 TI - Immunohistologic study on the expressions of alpha-fetoprotein and protein induced by vitamin K absence or antagonist II in surgically resected small hepatocellular carcinoma. AB - Sixty-eight cases of single hepatocellular carcinoma (HCC) with less than 3 cm of diameter were immunohistochemically examined for the expressions of alpha fetoprotein (AFP) and protein induced by vitamin K absence or antagonist II (PIVKA-II). In cancerous tissues, the expression rate was significantly higher for PIVKA-II (34 cases [50%]) than AFP (21 cases [31%]) (P <.05), suggesting a higher specificity of PIVKA-II to small HCC. Sixteen of the 68 cases (24%) were positive to both AFP and PIVKA-II, and in 8 of the 16 cases, AFP and PIVKA-II expressing areas within a nodule were clearly divided by a fibrous septum. According to histologic grades, PIVKA-II expression was confirmed in 2 of the 15 well-differentiated HCCs, and in the well-differentiated component of 6 of the 12 "nodule-in-nodule"-type well-differentiated HCCs. AFP expression was not found in well-differentiated HCCs, but found in 16 of the 40 moderately differentiated HCCs (40%) and in the moderately differentiated component of 3 of the 12 "nodule in-nodule"-type well-differentiated HCCs. The positive rate in the tissues was correlated to the serum levels for both AFP and PIVKA-II. In addition, frequency of tissue-PIVKA-II expression was higher than tissue-AFP expression in the cases whose serum protein level was within the normal range. This indicates that AFP and PIVKA-II have different patterns of tissue expression and of secretion to the blood. In comparison with tissue-AFP-negative cases, tissue-AFP-positive HCCs had a larger tumor size, higher frequencies of portal vein invasion and intrahepatic metastasis, a high Ki-67 labeling index, and a lower rate of recurrence-free survival. Thus, tissue-AFP-positive HCCs are suggested to be biologically more malignant than those HCCs that are AFP-negative and PIVKA-II-positive. PMID- 11732003 TI - Spatiotemporal expression of angiogenesis growth factor receptors during the revascularization of regenerating rat liver. AB - Regenerating liver was evaluated for the spatiotemporal expression of angiogenic growth factor receptors on endothelial cell (EC) membranes during revascularization resulting from 70% partial hepatectomy (PHx). Fractions enriched in EC membranes were examined by Western blot for angiogenic growth factor receptor expression from 1 to 14 days after PHx. Increases in vascular endothelial growth factor (VEGF) receptors Flt-1 and Flk-1/KDR, angiopoietin receptors Tie-1, Tie-2, and platelet-derived growth factor receptor beta (PDGF Rbeta), modest increases in epidermal growth factor receptor (EGF-R), and no increase in hepatocyte growth factor receptor (c-Met) or fibroblast growth factor receptors (FGF-R) were observed in isolated membranes during EC proliferation. All receptors were tyrosine phosphorylated, and therefore activated, during peak expression. Immunofluorescence staining of regenerating liver identified populations with increased receptor expression, indicating cells receptive to ligand signaling. EGF-R was upregulated evenly throughout the sinusoidal membrane, whereas c-Met was observed on hepatocyte canaliculae, bile duct epithelium, and large vessel EC. Tie-2 and PDGF-Rbeta were increased on sinusoidal and large vessel EC, whereas Tie-1 was expressed in EC surrounding avascular hepatic islands. Flk-1/KDR was increased on large vessels with slight increases on sinusoidal EC, whereas Flt-1 was increased in arterioles, sinusoidal EC as well as in hepatocytes. Although Flt-1 was phosphorylated on isolated hepatocytes, vascular endothelial growth factor(165) (VEGF(165)) did not induce a proliferative or motogenic response. Proliferation assays on isolated EC indicated responsiveness to VEGF(165), but synergism among several growth factors including PDGF-BB was also observed. The data identify novel autocrine and paracrine interactions and indicate that each growth factor acts on a specific set of EC at specific times during revascularization of regenerating liver. PMID- 11732004 TI - Delivery of IkappaB superrepressor gene with adenovirus reduces early alcohol induced liver injury in rats. AB - Chronic alcohol administration increases gut-derived endotoxin in the portal blood, which activates Kupffer cells through nuclear factor kappaB (NF-kappaB) to produce toxic mediators such as proinflammatory cytokines, leading to liver injury. Therefore, a long-term intragastric ethanol feeding protocol was used here to test the hypothesis that NF-kappaB inhibition would prevent early alcohol induced liver injury. Adenoviral vectors encoding either the transgene for IkappaB superrepressor (AdIkappaB-SR) or the bacterial beta-galactosidase reporter gene (AdlacZ) were administered intravenously to Wistar rats. Animals were fed a high-fat liquid diet with either ethanol or isocaloric maltose-dextrin (control) for 3 weeks. There was no significant difference in mean urine alcohol concentrations between the groups fed ethanol. IkappaB-SR expression was increased for up to 2 weeks after injection, but was undetectable at 3 weeks. NF kappaB activation was increased by ethanol and associated with up-regulation of tumor necrosis factor alpha (TNF-alpha). These increases were blunted significantly up to 2 weeks by AdIkappaB-SR. Dietary alcohol significantly increased liver to body weight ratios and serum alanine transaminase (ALT) levels in AdlacZ-treated animals, effects that were blunted significantly in AdIkappaB SR-treated rats. Ethanol caused severe steatosis, inflammation, and focal necrosis in AdlacZ-treated animals. These pathologic changes were significantly decreased by AdIkappaB-SR. The protective effects of IkappaB-SR were significant 2 weeks after injection, but were lost at 3 weeks when IkappaB-SR was no longer expressed. Ethanol increased 4-hydroxynonenal as a maker of oxidative stress in both AdlacZ and AdIkappaB groups. These data support the hypothesis that NF kappaB inhibition prevents early alcohol-induced liver injury even in the presence of oxidative stress. PMID- 11732005 TI - Gene expression of tumor necrosis factor alpha and TNF-receptors, p55 and p75, in nonalcoholic steatohepatitis patients. AB - The main objective of this study was to analyze the pathogenic role of the tumor necrosis factor alpha (TNF-alpha) system in the development of nonalcoholic steatohepatitis (NASH). Fifty-two obese patients were studied. We investigated: (1) the expression of mRNA of TNF-alpha and their p55 and p75-receptors by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) in hepatic and adipose tissues; and (2) the relationship between TNF-alpha, p55, and p75 and the severity of NASH. Obese patients without NASH were the control group. A remarkable increase in the expression of mRNA of TNF-alpha was found in patients with NASH in hepatic tissue (0.65 +/- 0.54) and in peripheral fat (0.43 +/- 0.45); in the control samples, the mRNA expression was 0.28 +/- 0.32, P <.007, and 0.26 +/- 0.22, P <.018, respectively. Furthermore, we found a significant increase in the mRNA levels of p55 receptor (2.42 +/- 1.81 vs. 1.56 +/- 1.17; P <.05); however, the mRNA expression of the p75 receptor was similar in both patients. Those patients with NASH with significant fibrosis presented an increase in the expression of mRNA TNF-alpha in comparison with those with a slight or nonexistent fibrosis. An overexpression of TNF-alpha mRNA is found in the liver and in the adipose tissue of NASH patients. The levels of mRNA-p55 are increased in the liver tissue of NASH patients. This overexpression is more elevated in patients with more advanced NASH. These findings suggest that the TNF alpha system may be involved in the pathogenesis of NASH. PMID- 11732006 TI - Adenosine monophosphate-activated protein kinase mediates the protective effects of ischemic preconditioning on hepatic ischemia-reperfusion injury in the rat. AB - Hepatic ischemia-reperfusion (I/R) injury associated with liver transplantation and hepatic resections are an unresolved problem in the clinical practice. Preconditioning is known to preserve energy metabolism in liver during sustained ischemia, but the molecular mechanisms underlying this effect are still unclear. Different metabolic signals, including adenosine monophosphate (AMP) and nitric oxide (NO), have been implicated in preconditioning. AMP-activated protein kinase (AMPK) protects cells by acting as a low-fuel warning system, becoming switched on by adenosine triphosphate (ATP) depletion. NO synthesis is induced by AMPK in the heart during ischemia. The aim of this study was to investigate: 1) whether preconditioning induces AMPK activation; and 2) if AMPK activation leads to ATP preservation and reduced lactate accumulation during prolonged ischemia and its relationship with NO. Preconditioning activated AMPK and concomitantly reduced ATP degradation, lactate accumulation, and hepatic injury. The administration of an AMPK activator, AICAR, before ischemia simulated the benefits of preconditioning on energy metabolism and hepatic injury. The inhibition of AMPK abolished the protective effects of preconditioning. The effect of AMPK on energy metabolism was independent of NO because the inhibition of NO synthesis in the preconditioned group and the administration of the NO donor before ischemia, or to the preconditioned group with previous inhibition of AMPK, had no effect on energy metabolism. Both preconditioning and AICAR pretreatment, through AMPK activation, may be useful surgical and pharmacologic strategies aimed at reducing hepatic I/R injury. PMID- 11732007 TI - Disturbances in hepatic cell-cycle regulation in mice with assembly-deficient keratins 8/18. AB - Simple epithelial tissues such as liver and pancreas express keratins 8 (K8) and 18 (K18) as their major intermediate filament proteins. K8 and K18 null mice and transgenic mice that express mutant K18 (K18C) manifest several hepatocyte abnormalities and demonstrate that K8/18 are important in maintaining liver tissue and cell integrity, although other potential functions remain uncharacterized. Here, we report an additional abnormal liver phenotype, which is similar in K8 null, K18 null, and K18C mouse models. Liver histologic examination showed large polynuclear areas that lacked cell membranes, desmosomal structures, and filamentous actin. Similar, but less prominent, areas were observed in the pancreas. The parenchyma outside the polynuclear areas displayed irregular sinusoidal structures and markedly enlarged nuclei. Most K8 null hepatocytes were positive for the proliferating cell nuclear antigen (PCNA) with a doubled DNA content in comparison with the predominantly PCNA-negative wild-type hepatocytes. The distribution of the 14-3-3zeta protein was also altered in K8 null mice. Taken together, our results indicate that absence of keratin filaments causes disturbances in cell-cycle regulation, driving cells into the S-G2 phase and causing aberrant cytokinesis. These effects could stem from disturbed functions of K8/18-dependent cell-cycle regulators, such as the signaling integrator, 14-3 3. PMID- 11732008 TI - Notch receptor expression in adult human liver: a possible role in bile duct formation and hepatic neovascularization. AB - Notch signaling is an evolutionarily conserved mechanism, used to regulate cell fate decisions. Four Notch receptors have been identified in man (Notch-1 to -4). In this study, semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to examine the expression pattern of Notch receptor genes in whole adult human liver and isolated liver cell preparations. All 4 receptors were expressed in the adult liver, with no significant differences in the levels of Notch-1, -2, and -4 messenger RNA (mRNA) between normal and diseased liver. However, Notch-3 expression appeared to be increased in diseased tissue. The distribution of Notch-1 and -4 in normal tissue was similar, with Notch-1 also detectable at low levels in the sinusoidal endothelium. Notch-2 expression was more widely distributed, and detectable in hepatocytes, medium-sized bile ducts, and the sinusoidal endothelium. Notch-3 expression was seen on hepatocytes, with weaker expression detectable in portal veins, hepatic arteries, and the sinusoids. In normal liver tissue Notch-1, -2, and -3 were found to be coexpressed on bile duct epithelium; however, with the exception of Notch-3 in primary sclerosing cholangitis (PSC) livers, expression was absent on proliferating ductules in all disease states examined. Interestingly, the expression of Notch-2 and -3 was associated with numerous small vessels within the portal tract septa of diseased tissue. The absence of Notch receptor expression on proliferating bile ductules and its presence on neovessels suggests that Notch signaling may be important for normal bile duct formation and the aberrant neovascularization seen in diseased liver tissue. PMID- 11732009 TI - The influence of human immunodeficiency virus coinfection on chronic hepatitis C in injection drug users: a long-term retrospective cohort study. AB - In this study we analyzed the influence of human immunodeficiency virus (HIV) infection on the course of chronic hepatitis C through multivariate analysis including age, alcohol consumption, immune status, and hepatitis C virus (HCV) related virologic factors. Eighty HIV-positive and 80 HIV-negative injection drug users included between 1980 and 1995 were matched according to age, gender, and duration of HCV infection and followed-up during 52 months. The progression to cirrhosis was the primary outcome measure. The impact of HIV on HCV-RNA load, histologic activity index, response to interferon therapy, and liver-related death was also considered. In HIV-positive patients, chronic hepatitis C was characterized by higher serum HCV-RNA levels (P =.012), higher total Knodell score (P =.011), and poorer sustained response to interferon therapy (P =.009). High serum HCV-RNA level was associated with low CD4-lymphocyte count (P =.001). Necroinflamatory score was higher in HIV-positive patients (P =.023) independently of the CD4-lymphocyte count, whereas increased fibrosis was related to decreased CD4-lymphocyte count (P =.011). The progression to cirrhosis was accelerated in HIV-positive patients with low CD4 cell count (RR = 4.06, P =.024) and in interferon-untreated patients (RR = 4.76, P =.001), independently of age at HCV infection (P =.001). Cirrhosis caused death in 5 HIV-positive patients. The risk of death related to cirrhosis was increased in heavy drinkers (RR = 10.8, P =.001) and in HIV-positive patients with CD4 cell count less than 200/mm(3) (RR = 11.9, P =.007). In this retrospective cohort study, HIV coinfection worsened the outcome of chronic hepatitis C, increasing both serum HCV-RNA level and liver damage and decreasing sustained response to interferon therapy. Age and alcohol were cofactors associated with cirrhosis and mortality. Interferon therapy had a protective effect against HCV-related cirrhosis no matter what the patient's HIV status was. PMID- 11732010 TI - Prevalence and risk factors for hepatitis C virus infection at an urban Veterans Administration medical center. AB - This study was designed to determine the seroprevalence and risk factors for hepatitis C virus (HCV) infection in veterans. Anti-HCV testing was performed in 1,032 patients and a questionnaire regarding sociodemographic characteristics and potential risk factors was administered. Adjusted prevalence of unique HCV positive patients using outpatient services was 17.7% (95% confidence interval [CI] 17.2%, 18.2%). The following risk factors were associated with HCV infection: a history of injection drug use (IDU), receipt of blood transfusion prior to 1992, history of tattoo (odds ratio [OR], 2.93; 95% CI, 1.70-5.08), combat job as a medical worker (OR, 2.68; 95% CI, 1.25-5.60), history of incarceration over 48 hours (OR, 2.56; 95% CI, 1.52-4.32), greater than 15 lifetime sexual partners (OR, 1.61; 95% CI, 0.94-2.76) and sexual relations with a prostitute (OR, 0.46; 95% CI, 0.25-0.82). We concluded that HCV is common in veterans. Risk factors independently associated with infection are IDU, prior transfusion, prior tattoo, combat medical work, incarceration, and multiple opposite sex partners. Infection with HCV among veterans is strongly associated with traditional risk factors for infection and less strongly associated with combat-related risk. PMID- 11732011 TI - Different hepatitis C virus nonstructural protein 3 (Ns3)-DNA-expressing vaccines induce in HLA-A2.1 transgenic mice stable cytotoxic T lymphocytes that target one major epitope. AB - The immunogenicity of the Hepatitis C virus (HCV) nonstructural protein 3 (NS3) was investigated using different DNA-based strategies and a preclinical mouse model transgenic for the HLA-A2.1 molecule. Plasmids expressing NS3 either as a wild-type protein, as a fusion with murine lysosome-associated-membrane protein-1 specific sequences, or under the control of the Semliki Forest virus replicase were evaluated in vitro and in vivo. All plasmids were shown to express the expected size protein. These 3 NS3-expressing vaccines induced overall comparable levels of CTLs when measured at different times postvaccination although mice injected with the NS3-LAMP expressing plasmid showed a particularly homogeneous and overall vigorous response (specific lysis ranged from 60% to 90 % for an E:T ratio of 33.3:1 with a mean CTL precursor frequency of 1:2.10(5) cells). Out of the four HLA-A2.1-restricted NS3 epitopes previously described in HCV infected patients (aa 1073-1081, aa 1406-1415; aa 1169-1177 and aa 1287-1296), the NS3-DNA generated CTLs were predominantly targeted at the aa 1073-1081 epitope. Peptide based immunization showed that the mouse repertoire was intact for all epitopes tested except one (aa 1287-1296). In conclusion, the 3 NS3-DNA vaccines although based on different mode of action, shared a comparable efficacy at inducing CTL. Surprisingly, the breadth of such response was restricted to a single, major epitope. PMID- 11732012 TI - Hepatitis B virus X protein transactivates inducible nitric oxide synthase gene promoter through the proximal nuclear factor kappaB-binding site: evidence that cytoplasmic location of X protein is essential for gene transactivation. AB - Nitric oxide appears to play a central role in the pathogenesis of many inflammatory disorders. We have previously shown that there is an enhanced intrahepatic expression of the inducible nitric oxide synthase (iNOS) gene during chronic hepatitis B virus infection, and that viral X protein (HBx) transcriptionally activates this cellular gene, but the molecular basis for this activation remains to be defined. We aimed to explore the involvement of different cis-acting elements of the human iNOS promoter in the HBx-mediated up regulation as well as the effect of the intracellular distribution of the HBx on its transacting function. Cotransfection of human hepatocyte-derived cells with wild-type or mutated iNOS promoter and with an HBx expression vector showed that functional inactivation of the proximal nuclear factor kappaB (NF-kappaB)-binding site of the iNOS promoter markedly reduced the HBx-mediated transcriptional activity. Mobility shift assays showed increased DNA-protein complexes comprising mainly the p50 and p65 members of NF-kappaB family in the nuclear extracts from HBx-transfected cells. Transient transfection experiments with HBx-expressing plasmids containing distinct cellular localization sequences showed that cytoplasmic location of this viral protein activated the iNOS promoter but when HBx was targeted to the nucleus, the HBx-induced luciferase activity was almost completely abrogated. In conclusion, cytoplasmic location of HBx protein is essential for the transcriptional activation of the iNOS gene through the nuclear translocation of p50-p65 heterodimers. PMID- 11732013 TI - Chronic hepatitis B. PMID- 11732015 TI - International hepatology. PMID- 11732014 TI - New challenge of hepatorenal syndrome: prevention and treatment. AB - Hepatorenal syndrome (HRS) remains one of the major therapeutic challenges in hepatology today. The pathogenesis is complex, but the final common pathway seems to be that sinusoidal portal hypertension, in the presence of severe hepatic decompensation, leads to splanchnic and systemic vasodilatation and decreased effective arterial blood volume. Renal vasoconstriction increases concomitantly, renal hemodynamics worsens, and renal failure occurs. Renal failure was shown 15 years ago to be potentially reversible after liver transplantation. This potential reversibility together with increased understanding of the pathogenesis has led to successful preliminary attempts to reverse HRS nonsurgically with combinations of splanchnic vasoconstrictors and colloid volume expansion, insertion of transjugular intrahepatic portovenous shunt radiologically, and improved forms of dialysis. Recent classification of HRS into the acute onset or severe type 1 with virtually 100% mortality and the more insidious less severe type II promises to shed more light on the pathogenesis of HRS, especially on the currently unrecognized precipitating factors. It is hoped that this classification will be included in the necessary and carefully performed clinical trials, which should lead to clearer indications for the available therapies. The challenge now is to use all this information to improve our management of cirrhotic patients to prevent occurrence of HRS in the future. PMID- 11732016 TI - A new player in the team: SOCS-3 socks it to cytokine signaling in the regenerating liver. PMID- 11732018 TI - Repair of orbital floor fractures with bioactive glass implants. AB - PURPOSE: The ideal management of orbital floor fractures has been highly controversial. Many implants, both autogenous and alloplastic, have been used to span the defects. This study evaluated the use of bioactive glass implants (BAG implant, S53P4; Abmin Technologies Ltd, Turku, Finland) for the repair of orbital floor defects caused by blunt facial trauma. PATIENTS AND METHODS: This retrospective review of 36 patients was carried out from 1995 to 1999. All patients were diagnosed with an orbital floor fracture or a large orbital blowout fracture. The BAG-implant was placed over the defect, using a subciliary or transconjunctival approach. No screw fixation was used when the implant was the correct size. Follow-up examination was done at 1 and 3 months after surgery. Twenty-eight (82%) of the patients were also seen at one-year follow-up (21 men and 7 women). RESULTS: The implants did not cause a foreign body reaction in the bone or soft tissue. There was no sign of resorption or infection, nor postoperative extrusion, hemorrhage, or displacement of the implant. Diplopia was seen preoperatively in 17 cases (61%) and postoperatively in 5 cases (18%). In 1 patient, the implant was removed 3 months after operation because of diplopia. Infraorbital nerve paresthesia was seen preoperatively in 9 patients (32%) and postoperatively in 5 patients (18%). The functional and cosmetic results were good at the 1-year follow-up. CONCLUSION: The BAG-implant is a well-tolerated material in orbital floor reconstruction. It provides a favorable environment for an uncomplicated healing process because it is bioactive and biocompatible and because it causes new bone formation. PMID- 11732020 TI - Repair of traumatic inferior orbital wall defects with nasoseptal cartilage. AB - PURPOSE: This study evaluated the effectiveness of nasoseptal cartilage for repairing traumatic orbital floor defects. PATIENTS AND METHODS: Autogenous septal cartilage was used in 20 patients. They were evaluated for the presence or absence of diplopia, enophthalmus, infraorbital nerve paresthesia, and ocular motility disorders. Surgical indications for orbital exploration included entrapment of orbital tissues, large orbital defect (greater than 50% of the orbital floor or more than 8 mm), or orbital floor defects with involvement of other zygomaticofrontal complex fractures. RESULTS: All patients were successfully treated by restoration of the orbital wall continuity. Follow-up at 1 week to 6 months showed 1 patient with postoperative enophthalmos and 1 patient with lower lid edema. There were no donor site and graft infections or graft extrusion. CONCLUSIONS: Nasal septal cartilage is a readily accessible autogenous tissue that should be considered when an autogenous graft is needed for orbital floor defect reconstruction. PMID- 11732022 TI - The osseointegration of Mitek mini anchors in the mandibular condyle. AB - PURPOSE: This study examined the histomorphology of the bone around the Mitek mini anchors (Mitek Inc, Norwood, MA) after 3 to 59 months of implantation. MATERIALS AND METHODS: Thirteen condylar specimens from 8 patients were evaluated. Each of these patients had had previously placed Mitek mini anchors and braided polyester sutures for temporomandibular joint (TMJ) articular disc stabilization. Histologic evaluation of each specimen was performed to determine the degree of osseointegration. Each specimen was also evaluated for anchor configuration, fracture, and particulation and the presence or absence of intervening fibrous connective tissue or an inflammatory cell infiltrate. RESULTS: All anchors became osseointegrated without evidence of associated inflammation or fibrous connective tissue formation along the surface. There was no evidence of anchor fracture or particulation. CONCLUSIONS: The Mitek mini anchor can osseointegrate in the mandibular condyle. These suture anchors remain intact and biocompatible for as long as 59 months, the longest interval examined in this study. PMID- 11732024 TI - Speech outcome after closure of oronasal fistulas with bone grafts. AB - PURPOSE: The purpose of this prospective study was to evaluate the outcome of speech after complete closure of oronasal fistulas with bone grafts and to determine the possible relationship between outcome of speech and the size and location of the oronasal fistulas. PATIENTS AND METHODS: Ten unilateral cleft lip and palate patients with postoperative oronasal fistulas, ranging in age from 7 to 14 years, underwent secondary alveolar cleft repair and closure of the oronasal fistulas with an iliac bone graft. All patients underwent videofluoroscopic evaluation of the velopharyngeal valve, audiologic assessment, and speech evaluation (resonance, nasal emission, articulation, intelligibility, and nasalance) preoperatively. The examinations were repeated 3 months postoperatively. RESULTS: Six patients had preoperative velopharyngeal competency (60%). Of the 4 patients with slight to mild velopharyngeal incompetency preoperatively, 2 developed velopharyngeal competency postoperatively. All patients had satisfactory audiologic function preoperatively. Every patient also was intelligible before and after surgery. Eight patients (80%) showed nasal emission before surgery and 7 of these patients improved postoperatively (P <.01). Nine patients had articulation errors before surgery, with no significant improvement postoperatively. Nasalance was significantly improved in selected sequences. All patients had variable levels of nasality preoperatively; 8 showed a significant decrease of nasality postoperatively (P <.002). The results were not related to location or size of the oronasal fistulas. CONCLUSION: A significant improvement in speech is noticeable after closure of oronasal fistulas. Early oronasal fistula closure might prevent permanent speech distortions acquired by the cleft palate patients at an early age. PMID- 11732026 TI - Postoperative antibiotic prophylaxis in mandibular fractures: A preliminary randomized, double-blind, and placebo-controlled clinical study. AB - PURPOSE: This study evaluated the difference between the effect of a 5-day postoperative course of oral antibiotics and a placebo on the incidence of postoperative infection in uncomplicated fractures of the mandible. PATIENTS AND METHODS: A prospective, randomized, double-blind clinical study using a placebo control was carried out. Thirty patients were randomly assigned into 2 groups: group 1 (14 patients) and group 2 (16 patients). Each group received penicillin G, 2 million U intravenously, every 4 hours through the preoperative period, intraoperative period, and for 12 hours postoperatively. Group 1 then received penicillin VK, 500 mg every 6 hours for 5 days. Group 2 received oral placebo using the same schedule for the same duration as group 1. The patients were than evaluated for evidence of infection 1, 2, 4, and 6 weeks postoperatively. RESULTS: Two of 14 patients in group 1 (14.3%) and 2 of 16 patients in group 2 (12.5%) developed infections. No statistically significant difference in the incidence of infection was noted between the groups. CONCLUSIONS: In this preliminary study, the use of postoperative oral antibiotics in uncomplicated fractures of the mandible had no benefit in reducing the incidence of infections. PMID- 11732028 TI - Eagle's syndrome: a report of 4 patients treated using a modified extraoral approach. AB - PURPOSE: This study evaluated a slightly modified extraoral approach to removing the symptomatic elongated styloid process. A brief description of this surgical technique and its comparison to the intraoral approach is provided. PATIENTS AND METHODS: Four patients in whom an extraoral approach was used to expose and remove the elongated calcified stylohyoid complex were evaluated. Five sides were treated and reviewed individually. All patients were interviewed preoperatively and postoperatively, and their records were thoroughly examined. Patients were objectively evaluated for pain, function, sensory and motor nerve deficits, and aesthetics. RESULTS: Follow-up time ranged from 6 to 39 months with an average of 21 months. Improvement in the original symptoms was reported for 4 of the 5 operated sides. Three of 5 procedures resulted in more than an 80% reduction in pain and improvement in function. All patients had a satisfactory cosmetic result and no paresthesia. CONCLUSIONS: The advantages of the extraoral approach are reflected in this small retrospective study by the absence of perioperative complications and the minimal blood loss, surgical morbidity, and hospital stay. PMID- 11732029 TI - Radiographic stereophotogrammetric evaluation of intersegmental stability after mandibular sagittal split osteotomy and rigid fixation. AB - PURPOSE: The purpose of this study was to determine if rigid fixation with bicortical screws and/or miniplates with monocortical screws prevent mobility at the osteotomy site after bilateral mandibular sagittal split osteotomy. PATIENTS AND METHODS: Three metal bone markers were inserted in the proximal and the distal segments of the mandible during the sagittal split operation in 10 patients. These served as measurement points in postoperative follow-up by radiographic stereophotogrammetry. The patients were examined at intervals during the first postoperative year. At each examination, 2 sets of radiographic stereograms were obtained: 1 in rest position and 1 with stress applied to the osteotomy sites. The difference in the position of the proximal segment in relation to the distal segment between the 2 sets of stereograms was recorded. Findings greater than 0.4 degrees and 0.2 mm change indicated true displacement of the bone segments. RESULTS: Immediately after surgery, mobility at of the osteotomy site(s) was found in 8 of 10 patients, and after 1 year it was still present in 4 patients. CONCLUSIONS: Fixation with bicortical screws or miniplates and monocortical screws does not prevent mobility at the osteotomy site after sagittal split osteotomies. This mobility may remain as long as 1 year after surgery. The term "rigid fixation" is thus not a proper term for this kind of fixation. PMID- 11732031 TI - A computer-based method for calculation of orbital floor fractures from coronal computed tomography scans. AB - PURPOSE: A computer program recently developed for the calculation of the orbital floor and fracture areas from coronal computed tomography (CT) scans was used in a study to evaluate the accuracy and ability of this new method. MATERIAL AND METHODS: The size of orbital floors and fabricated fractures in 14 dried, anatomic specimens were measured in coronal CT scans by 3 independent observers. Based on this data set, the orbital floor and fracture regions were calculated with the newly developed computer program. These calculated regions were then compared with a direct measurement of the specimens that had been obtained by digital photography. The accuracy of the computer-based calculations was assessed using Lin's concordance correlation coefficient. RESULTS: The size of the orbital floor (mean +/- SD) was found to be 5.21 +/- 0.39 cm(2) by direct measurement of the specimens and 5.30 +/- 0.52 cm(2) by calculation with the computer program. The region of the fracture (mean +/- SD) was 1.05 +/- 0.64 cm(2) by direct measurement and 1.01 +/- 0.62 cm(2) by computer calculation. The between-method mean difference (direct measurement minus computer based calculation) was -0.09 cm(2) (or 1.7% of mean orbital floor region) for orbital floor region and 0.04 cm(2) (or 3.8% of mean fracture region) for fracture region. CONCLUSIONS: This accurate and time-saving method is practicable for determining the size and location of orbital floor fractures. This calculation program can be advantageously applied in the clinical management of blowout fractures of the orbit. PMID- 11732032 TI - A model for temporomandibular joint disc repositioning surgery. AB - PURPOSE: The present study establishes a model of reversible anterior disc displacement using the rabbit craniomandibular joint (CMJ) and examines the period required for successful disc repositioning surgery. MATERIALS AND METHODS: The left CMJ was exposed surgically in 27 animals and a suture positioned at the posterolateral portion of the disc was pulled in an anterior direction causing 7 mm of anterior disc displacement. The suture was then fixed to the zygomatic arch. The changes in the CMJ were observed microscopically in 3 rabbits each at 2, 3, 5, 8, and 24 weeks. Another 12 rabbits underwent secondary disc repositioning surgery either the second and third week after disc displacement, and then 3 rabbits from each group were assessed microscopically at 5 to 6 weeks and 21 to 22 weeks after the first operation. Ten rabbits had a sham operation, and 2 served as unoperated controls. Histomorphometric evaluation included changes in morphology, cartilage, and disc condition. RESULTS: Progressive osteoarthritic change was induced by disc displacement. Articular disc perforation at 5 weeks after the first operation was accompanied by subchondral bone exposure. The CMJ morphology was almost normal in rabbits that underwent secondary disc repositioning 2 weeks after disc displacement. However, osteoarthritic changes and perforated discs similar to those in rabbits that did not undergo secondary disc repositioning were seen in animals in which repositioning was done in the third week. CONCLUSION: Artificial disc displacement in the rabbit causes osteoarthritic changes are reversible when the displaced disc is restored within after 2 weeks but not at 3 weeks. PMID- 11732033 TI - Influence of distraction rates on the temporomandibular joint position and cartilage morphology in a rabbit model of mandibular lengthening. AB - PURPOSE: Although various aspects of bone formation during distraction osteogenesis have been studied extensively, there are only limited experimental data concerning the influence of mandibular distraction rates on structural alterations in the temporomandibular joint (TMJ). In this study, a rabbit model of unilateral mandibular distraction was used to test the effects of various strain schedules on the position and morphology of the TMJ. MATERIAL AND METHODS: Fifty-two immature white female rabbits were used. The distraction procedure was performed using physiologic (2,000 microstrains, 1 per day) and elevated strain magnitudes (20,000 microstrains, 1 per day), as well as high strain magnitudes (200,000 and 300,000 microstrains, 1 per day). The investigation of the TMJ included clinical, radiologic, and histologic aspects. RESULTS: Clinical and radiologic examinations at the end of the distraction period showed no evidence of joint luxation even at maximal distraction rates. Histologic and ultrastructural analyses revealed a positive correlation between the degree of mechanical loading and the development of degenerative alterations in the cartilage. In samples distracted at hyperphysiologic strain magnitudes, all cartilaginous layers were reduced in the regions of the TMJ that had been exposed to the higher pressure forces. The fibrous layer became nearly completely destroyed. CONCLUSIONS: These experimental data show that distraction schedules with single but hyperphysiologic loads may lead to degenerative or even early arthrotic changes in the condyle. These data support the principle that distraction protocols should be performed without extensive mechanical loading on the TMJ. PMID- 11732035 TI - Traumatic hyphema: a comprehensive review. PMID- 11732036 TI - Integration of the medical degree in oral and maxillofacial surgery: A 10-year follow-up. AB - PURPOSE: Ten years ago, the American Association of Oral and Maxillofacial Surgeons membership was surveyed to evaluate the influence that integration of a medical degree might have on oral and maxillofacial training. The intent of the current survey was to reassess the influence a medical degree has had on the specialty over the past 10 years. PATIENTS AND METHODS: The effects of a medical degree on privileges, referral patterns, and its role in the office, hospital, and academic settings were studied. Biographical data was collected and responses were evaluated for the following groups: 1) the nation as a whole, 2) MD versus non-MD oral and maxillofacial surgeons (OMS), 3) geographic regions of practice, 4) population, 5) number of years in practice, and 6) involvement in academic programs. RESULTS: The results of this survey were similar to the previous one. Substantially different responses were seen between the dual-degree and single degree OMS as well as differences between geographic locations, years in practice, and academic involvement. MD-DDS and academic OMS again possessed a broader spectrum of privileges than their colleagues. Recently trained OMS again possessed a greater number of privileges than more experienced surgeons. CONCLUSION: Overall there has been a trend toward increasing surgical privileges over the past 10 years for both single- and dual-degree OMS. The results presented emphasize the need to continually assess the influence that a medical degree has on the specialty. PMID- 11732039 TI - Endoscopic-assisted reduction and fixation of a mandibular subcondylar fracture: report of a case. PMID- 11732040 TI - Villonodular synovitis of the temporomandibular joint: case report. PMID- 11732041 TI - Spontaneous remission of a solitary eosinophilic granuloma of the mandible after biopsy: a case report. PMID- 11732042 TI - Myocardial ischemia and delayed recovery after anesthesia in a patient with Cockayne syndrome: a case report. PMID- 11732043 TI - Maxillary distraction osteogenesis for cleft lip and palate children using an external, adjustable, rigid distraction device: a report of 2 cases. PMID- 11732044 TI - Bilateral lingual nerve injury after laryngoscopy for intubation. PMID- 11732045 TI - The use of a pedicled buccal fat pad graft for bone coverage in primary palatorrhaphy: a case report. PMID- 11732046 TI - Epithelial-myoepithelial carcinoma of the palate: a case report. PMID- 11732047 TI - Variations in lower lip reconstruction. PMID- 11732048 TI - Intramuscular hemangioma with phleboliths or choristoma? PMID- 11732049 TI - Infected thyroglossal duct cyst or "the President's syndrome" (lingual abscess). PMID- 11732050 TI - Identification of a novel plant-specific kinesin-like protein that is highly expressed in interphase tobacco BY-2 cells. AB - Through reverse transcription-polymerase chain reaction and Northern blot analysis, we identified TBK5, a novel plant-specific kinesin-like protein (KLP) that is highly expressed in interphase tobacco BY-2 cells. TBK5 mRNA was present at a high level throughout the growth cycle, even in cells that had entered the stationary phase, where cell proliferation had ceased. However, transcripts for five other tobacco KLPs that we have identified were preferentially expressed in mitotic cells, and either not or only slightly accumulated in cells that had entered the stationary phase. Thus, TBK5 appears to be a KLP whose cellular function most closely relates to the cortical array of microtubules that plays a key role in plant cell morphogenesis. The predicted structure of TBK5 is characterized by a central motor domain that is phylogenetically distant from those of other reported KLPs, coiled-coil domains located on both sides of the motor domain, and a basic C-terminal domain. In addition, TBK5 has a putative neck domain which is closely related to the neck domain of KLPs with C-terminal motor domains, previously shown to control the direction of KLP movement towards the minus ends. Antibodies against truncated TBK5 recognized a polypeptide with a molecular mass of 74 kDa in cytoplasmic extracts of interphase cells, and this polypeptide cosedimented with microtubules assembled in the cytoplasmic extracts. The 74 kDa polypeptide corresponding to TBK5 dissociated from microtubules with high concentrations of NaCl but was not dissociated by MgATP. We hypothesize that TBK5 functions in the regulation of the arrangement of cortical microtubules. PMID- 11732051 TI - Morphology of rsw1, a cellulose-deficient mutant of Arabidopsis thaliana. AB - The rsw1 mutant of Arabidopsis thaliana is mutated in a gene encoding a cellulose synthase catalytic subunit. Mutant seedlings produce almost as much cellulose as the wild type at 21 degrees C but only about half as much as the wild type at 31 degrees C. We used this conditional phenotype to investigate how reduced cellulose production affects growth and morphogenesis in various parts of the plant. Roots swell in all tissues at 31 degrees C, and temperature changes can repeatedly switch them between swollen and slender growth patterns. Dark-grown hypocotyls also swell, whereas cotyledons and rosette leaf blades are smaller, their surfaces are more irregular and their petioles shorter. Leaf trichomes swell and branch abnormally. Plants readily initiate inflorescences at 31 degrees C which have shorter but not fatter bolts and stomata which bulge above the uneven surface of internodes. Bolts carry the normal number of flowers, but their stigmas protrude beyond the shortened sepals and petals. Anthers dehisce normally, but self-fertilisation is reduced because the stigma is well above the anthers. Anther filaments are short and show a crumpled surface. Viable pollen develops, but female reproductive competence and postpollination development are severely impaired. We conclude that the RSW1 gene is important for cellulose synthesis in many parts of the plant and that reduced cellulose synthesis suppresses organ expansion rather than organ initiation, causes radial swelling only in the root and hypocotyl, but makes the surfaces of many organs uneven. We discuss some possible reasons to explain why different organs vary in their responses. The morphological changes suggest that RSW1 contributes cellulose to primary walls but do not yet exclude a role during secondary-wall deposition. PMID- 11732052 TI - NaAGP4 is an arabinogalactan protein whose expression is suppressed by wounding and fungal infection in Nicotiana alata. AB - Arabinogalactan proteins (AGPs) are proteoglycans secreted by plant cells that have been implicated in plant growth and development. Most AGPs cloned to date possess highly labile glycosylphosphatidylinositol (GPI) lipid anchors. These anchors transiently attach AGPs to the plasma membrane before they are released into the cell wall following GPI anchor hydrolysis. We have isolated and partially sequenced the protein core of an AGP purified from styles of Nicotiana alata. The protein sequence data were utilised to clone the AGP's gene, NaAGP4. This AGP shares about 78% sequence identity with the tomato AGP LeAGP-1. RNA gel blot analyses of different plant organs indicate that NaAGP4 is expressed in the same tissues and at similar levels as LeAGP-1. Furthermore, NaAGP4 like LeAGP-1 is rapidly suppressed by tissue wounding and by pathogen infection. We believe NaAGP4 and LeAGP-1 are the first described examples of orthologous AGPs from different plant species. In contrast, another AGP from N. alata, NaAGP1, is comparatively unaffected by wounding and pathogen infection, although this AGP is expressed in similar tissues and at similar levels as NaAGP4. PMID- 11732053 TI - Cell biology and genetics of root hair formation in Arabidopsis thaliana. AB - In this review we integrate the information available on the cell biology of root hair formation with recent findings from the analysis of root hair mutants of Arabidopsis thaliana. The mature Arabidopsis root epidermis consists of root-hair producing cells and non-root-hair-producing cells. Root hair growth begins with a swelling of the outer epidermal wall. It has been postulated that this is due to a pH-mediated localised cell wall loosening. From the bulge a single root hair emerges which grows by tip growth. The root hair tip consists of a vesicle-rich zone and an organelle-rich subapical zone. The vesicles supply new plasma membrane and cell wall material for elongation. The cytoskeleton and its associated regulatory proteins such as profilin and spectrin are proposed to be involved in the targeting of vesicles. Ca2+ influxes and gradients are present in hair tips, but their function is still unclear. Mutants have been isolated with lesions in various parts of the root hair developmental pathway from bulge identity and initiation to control of tip diameter and extent and polarity of elongation. PMID- 11732054 TI - On the alignment of cellulose microfibrils by cortical microtubules: a review and a model. AB - The hypothesis that microtubules align microfibrils, termed the alignment hypothesis, states that there is a causal link between the orientation of cortical microtubules and the orientation of nascent microfibrils. I have assessed the generality of this hypothesis by reviewing what is known about the relation between microtubules and microfibrils in a wide group of examples: in algae of the family Characeae, Closterium acerosum, Oocystis solitaria, and certain genera of green coenocytes and in land plant tip-growing cells, xylem, diffusely growing cells, and protoplasts. The salient features about microfibril alignment to emerge are as follows. Cellulose microfibrils can be aligned by cortical microtubules, thus supporting the alignment hypothesis. Alignment of microfibrils can occur independently of microtubules, showing that an alternative to the alignment hypothesis must exist. Microfibril organization is often random, suggesting that self-assembly is insufficient. Microfibril organization differs on different faces of the same cell, suggesting that microfibrils are aligned locally, not with respect to the entire cell. Nascent microfibrils appear to associate tightly with the plasma membrane. To account for these observations, I present a model that posits alignment to be mediated through binding the nascent microfibril. The model, termed templated incorporation, postulates that the nascent microfibril is incorporated into the cell wall by binding to a scaffold that is oriented; further, the scaffold is built and oriented around either already incorporated microfibrils or plasma membrane proteins, or both. The role of cortical microtubules is to bind and orient components of the scaffold at the plasma membrane. In this way, spatial information to align the microfibrils may come from either the cell wall or the cell interior, and microfibril alignment with and without microtubules are subsets of a single mechanism. PMID- 11732056 TI - Wall architecture in the cellulose-deficient rsw1 mutant of Arabidopsis thaliana: microfibrils but not microtubules lose their transverse alignment before microfibrils become unrecognizable in the mitotic and elongation zones of roots. AB - The rsw1 mutant of Arabidopsis thaliana has a single amino acid substitution in a putative glycosyl transferase that causes a temperature-dependent reduction in cellulose production. We used recently described methods to examine root growth by surface marker particles, cell wall structure by field emission scanning electron microscopy and microtubule alignment by immunofluorescence after the mutant is transferred to its restrictive temperature. We find that raising the temperature quickly accelerates root elongation in both wild type and mutant, presumably as a result of general metabolic stimulation, but that in the mutant, the rate declines within 7-8 h and elongation almost ceases after 24 h. Radial swelling begins at about 6 h in the mutant and root diameter continues to increase until about 24 h. The normal transverse alignment of microfibrils is severely impaired in the mutant after 8 h, and chemical inhibition of cellulose synthesis by 2,6-dichlorobenzonitrile causes a similar loss of orientation. After 24 h, microfibrils are not clearly visible in the walls of cells that would have been in the mitotic and early-elongation zone of wild-type roots. Changes in older cells are less marked; loss of transverse microfibril orientation occurs without disruption to the transverse orientation of cortical microtubules. The wild type shows none of the changes except for acceleration of elongation, which in its case is sustained. We conclude that microfibril alignment requires the normal functioning of RSW1 and that, in view of the effects of dichlorobenzonitrile, there may be a more general linkage between the rate of cellulose production and its proper alignment. PMID- 11732055 TI - Pac-Man does not resolve the enduring problem of anaphase chromosome movement. AB - The Pac-Man hypothesis suggests that poleward movement of chromosomes during anaphase A is brought about by: disassembly of kinetochore microtubules (MTs) at the kinetochore; generation of the poleward force exclusively at or very close to the kinetochore; and the required energy coming from coupled disassembly of these MTs. This model has become widely accepted and cited as the sole or major mechanism of anaphase A. Rarely acknowledged are several significant phenomena that refute some or all of these postulates. We summarise these anomalies as follows: poleward movement of chromosomes occurring without insertion of any MTs at the kinetochore; "anaphase" shortening of kinetochore fibres in spindles entirely devoid of chromosomes and, presumably, kinetochores; continued movement of chromosomes while their severed kinetochore stub elongated poleward after treatment with UV microbeams; and fluxing of tubulin subunits through kinetochore MTs during anaphase A, indicating that during anaphase, kinetochore MTs disassemble partly or solely at the poles. PMID- 11732057 TI - Cytoplasmic acidification with butyric acid does not alter the ionic conductivity of plasmodesmata. AB - The effect of lowering cytoplasmic pH on the ionic conductivity of higher-plant plasmodesmata was investigated with corn (Zea mays L. cv. Black Mexican Sweet) suspension culture cells. Exposure to butyric acid decreased the cytoplasmic pH by 0.8 units. Intercellular communication was monitored by electrophysiological techniques that allowed the measurement of membrane resistances of sister cells and the electrical resistance of the plasmodesmata connecting them. The decrease in cytoplasmic pH did not affect the resistance of plasmodesmata, despite the fact that the butyric acid treatment more than doubled the concentration of cytoplasmic calcium. This is discussed in light of previous findings that increases in cytoplasmic calcium increase the electrical resistance of plasmodesmata. PMID- 11732058 TI - Wall ingrowth architecture in epidermal transfer cells of Vicia faba cotyledons. AB - We describe the use of scanning electron microscopy to provide novel views of the three-dimensional morphology of the ingrowth wall in epidermal transfer cells of cotyledons of developing Vicia faba seed. Wall ingrowth deposition in these cells amplifies the surface area of plasma membrane available for transport of solutes during cotyledon development. Despite the physiological importance of such amplification, little is known about wall ingrowth morphology and deposition in transfer cells. A detailed morphological analysis of wall deposition in this study clearly established for the first time that wall ingrowths are deposited at scattered, discrete loci as papillate ingrowth projections. The new views of the ingrowth wall revealed that these projections branch and fuse laterally, and fusion occurs by fine connections to form a fenestrated sheet or layer. This sheet of wall material then provides a base for further deposition of ingrowth projections to progressively build many interconnected, fenestrated layers. Consolidations, or filling-in, of the fenestrae in these layers appears to occur from small fingerlike protrusions of wall material which extend laterally from the most recently deposited surface of the fenestrae. We propose that deposition of fenestrated layers may provide a mechanism for maintaining continuous amplification of plasma membrane surface area in the face of turnover of the plasma membrane and transporter proteins associated with it. The techniques reported in this paper will provide new opportunities to investigate wall ingrowth deposition and its regulation in transfer cells. PMID- 11732059 TI - Haustoria in action: case studies of nitrogen acquisition by woody xylem-tapping hemiparasites from their hosts. AB - This review discusses studies conducted by the author and his colleagues on mistletoes and root hemiparasites native to Western Australia. Morphological characteristics of haustoria are described and their anatomical features are discussed in relation to uptake, transfer, and metabolism of xylem-borne nitrogenous solutes derived from a host. Experimental approaches used include comparisons of xylem sap composition of parasite and host(s), solute pool analyses and enzymatic properties of haustoria, host xylem feeding of 15N labelled solutes to follow the fate of label in haustoria and body of the parasite, and studies using species-specific nonprotein amino acids to validate successful uptake from hosts or occasional backflow of xylem-borne solutes to a host. Field studies on promiscuous root hemiparasites assess frequencies of exploitation of different hosts. 15N natural abundance assays of host and parasite dry matter demonstrate marked preference by Santalum acuminatum for N2 fixing as opposed to nonfixing hosts. The ability of Olax phyllanthi to continue to exploit deep-rooted hosts ranks of importance when xylem water potentials of other hosts go out of sucking range during periods of water stress. Comparisons of xylem sap composition of parasites feeding on different hosts indicate remarkable versatility by haustoria in uptake and utilization of the different major nitrogenous solutes received from these hosts. Solute pools in parasites partly reflect metabolic transformations accentuated by haustoria while also indicating direct throughput from xylem of a host. The review concludes by showing how empirically based modelling techniques can be used to estimate proportional gains of N by parasites from single hosts and repercussions on host growth which accompany such exploitation. PMID- 11732060 TI - Plasma membrane-cell wall connections: roles in mitosis and cytokinesis revealed by plasmolysis of Tradescantia virginiana leaf epidermal cells. AB - Tradescantia virginiana leaf epidermal cells were plasmolysed by sequential treatment with 0.8 M and 0.3 M sucrose. Plasmolysis revealed adhesion of the plasma membrane to the cell wall at sites coinciding with cytoskeletal arrays involved in the polarisation of cells undergoing asymmetric divisions--cortical actin patch--and in the establishment and maintenance of the division site- preprophase band of microtubules and filamentous (F) actin. The majority of cells retained adhesions at the actin patch throughout mitosis. However, only approximately 13% of cells formed or retained attachments at the site of the preprophase band. After the breakdown of the nuclear envelope, plasmolysis had a dramatic effect on spindle orientation, cell plate formation, and the plane of cytokinesis. Spindles were rotated at abnormal angles including tilted into the plane of the epidermis. Cell plates formed but were quickly replaced by vacuole like intercellular compartments containing no Tinopal-stainable cell wall material. This compartment usually opened to the apoplast at one side, and cytokinesis was completed by the furrow extending across the protoplast. This atypical cytokinesis was facilitated by a phragmoplast containing microtubules and F-actin. Progression of the furrow was unaffected by 25 micrograms of cytochalasin B per ml but inhibited by 10 microM oryzalin. Phragmoplasts were contorted and misguided and cytokinesis prolonged, indicating severe disruption to the guidance mechanisms controlling phragmoplast expansion. These results are discussed in terms of cytoskeleton-plasma membrane-cell wall connections that could be important to the localisation of plasma membrane molecules defining the cortical division site and hence providing positional information to the cytokinetic apparatus, and/or for providing an anchor for cytoplasmic F-actin necessary to generate tension on the phragmoplast and facilitate its directed, planar expansion. PMID- 11732061 TI - Motile tubular vacuoles in extramatrical mycelium and sheath hyphae of ectomycorrhizal systems. AB - Extramatrical mycelium and outer hyphae of the sheath of Eucalyptus pilularis Pisolithus tinctorius mycorrhizas contain abundant motile tubular vacuoles which accumulate the carboxyfluorescein analogue Oregon Green 488 carboxylic acid. The fluorochrome accumulates in a system of small vacuoles, tubules, and larger vacuoles, which are interlinked, motile, and pleiomorphic, in external hyphae, cords, and hyphae of the outer sheath. There is often a difference in fluorescence between two neighbouring cells, indicating that the dolipore septum exercises control on the movement of material between cells. Generally the motile tubular vacuole system in mycorrhizas resembles that previously found in isolated mycelium. The majority of fungal cells in the sheath contain no fluorochrome even after long exposure of the mycorrhiza to the solution, but with differential interference optics the cells are clearly seen to be alive and to contain vacuoles resembling those in the outer hyphae. In translocation experiments, long distance transport of the fluorochrome is slow and slight, or even nonexistent in some cases. PMID- 11732062 TI - Structure and expression of the genes encoding proteins resident in large peripheral vesicles of Phytophthora cinnamomi zoospores. AB - Zoospores of Phytophthora spp. contain several characteristic types of peripheral vesicles. One of these, large peripheral vesicles, has been proposed to act as a nutrient store and in P. cinnamomi has been shown to contain three immunologically related high-molecular-weight proteins, designated LPVs. We have used antibodies directed against P. cinnamomi zoospores and cysts to isolate several cDNA clones which are products of the Lpv genes and encode one or more of the LPV proteins present in large peripheral vesicles. Northern blot analysis demonstrated the presence of three large Lpv transcripts (11-14 kb) in RNA isolated from hyphae which had been induced to form sporangia. Coordinate accumulation of the three transcripts occurred after induction of sporangium formation: no transcript was observed in uninduced hyphae and maximum transcript levels of all three transcripts were seen 4-6 h after induction. Genomic Southern blots indicated that P. cinnamomi contains three Lpv genes, presumably corresponding to the three transcripts and proteins seen in Northern and Western blots, respectively. Partial genomic clones representing two of the Lpv genes were isolated and characterized by restriction mapping and partial DNA sequencing. In the regions sequenced, the genes were > 99% identical, the high degree of conservation extending at least 415 bp downstream of their polyadenylation sites. The Lpv coding regions contained a variable number (approximately 12-18) of highly conserved 534 bp repeats, flanked by apparently unique sequences. Variation in the number of repeats in the Lpv genes was responsible for the different sizes of the three transcripts and proteins. Database searches using the Lpv nucleotide and deduced amino acid sequences failed to detect any similar sequences. We discuss the molecular events which may have been involved in the evolution of the Lpv genes and the nature of the products of these genes. PMID- 11732063 TI - The cytoskeleton and spatial control of cytokinesis in the plant life cycle. AB - One of the intriguing aspects of development in plants is the precise control of division plane and subsequent placement of walls resulting in the specific architecture of tissues and organs. The placement of walls can be directed by either of two microtubule cycles. The better known microtubule cycle is associated with control of the future division plane in meristematic growth where new cells become part of tissues. The future daughter domains are determined before the nucleus enters prophase and the future site of cytokinesis is marked by a preprophase band (PPB) of cortical microtubules. The spindle axis is then organized in accordance with the PPB and, following chromosome movement, a phragmoplast is initiated in the interzone and expands to join with parental walls at the site previously occupied by the PPB. The alternative microtubule cycle lacks both the hooplike cortical microtubules of interphase and the PPB. Wall placement is determined by a radial microtubule system that defines a domain of cytoplasm either containing a nucleus or destined to contain a nucleus (the nuclear cytoplasmic domain) and controls wall placement at its perimeter. This more flexible system allows for cytoplasmic polarization and migration of nuclei in coenocytes prior to cellularization. The uncoupling of cytokinesis from karyokinesis is a regular feature of the reproductive phase in plants and results in specific, often unusual, patterns of cells which reflect the position of nuclei at the time of cellularization (e.g., the arrangement of spores in a tetrad, cells of the male and female gametophytes of angiosperms, and the distinctive cellularization of endosperm). Thus, both microtubule cycles are required for completion of plant life cycles from bryophytes to angiosperms. In angiosperm seed development, the two methods of determining the boundaries of domains where walls will be deposited are operative side by side. Whereas the PPB cycle drives embryo development, the radial-microtubule-system cycle drives the common nuclear type of endosperm development from the syncytial stage through cellularization. However, a switch to the PPB cycle can occur in endosperm, as it does in barley, when peripheral cells divide to produce a multilayered aleurone. The triggers for the switch between microtubule cycles, which are currently unknown, are key to understanding plant development. PMID- 11732064 TI - Membrane geometry of "open" prolamellar bodies. AB - The "open" type of prolamellar body in etiplasts was examined by electron microscopy to characterise its three-dimensional organisation. As in more compact forms of prolamellar body, its basic geometric unit is a tetrahedrally branched tubule. In the "open" type, these lie smoothly confluent with one another at the vertices of 5- and 6-membered rings which circumscribe the faces of three kinds of polyhedra: pentagonal dodecahedra (with 12 pentagonal faces), 14-hedra (2 opposite hexagonal faces joined by two circlets of six pentagonal faces), and 15 hedra (3 hexagonal and 12 pentagonal faces). These polyhedra join confluently in their turn, sharing faces with one another in at least one recognisable super structure which accounts for the appearance of "open" prolamellar bodies in many ultrathin sections. In this organisation, columns of pentagonal dodecahedra are arranged at 120 degrees to one another in the x-y-plane of the lattice. They do not fill the plane but intersect so as to delimit voids in the form of hexagonally arranged 14-hedra (with hexagonal rings in the x-y-plane). Strata of this type alternate with strata made of face-sharing 15-hedra (with their hexagonal rings normal to x-y), which also delimit 14-hedra. The 14-hedra thus lie in register in the z-axis in hexagonally arranged columns, normal to the alternating strata. Other possible organisations cannot be excluded and local variations and dislocations certainly occur, but many micrographs that display elements of symmetry in "open" prolamellar bodies can be matched to thin slices through such a model. Its geometry is like that of the cages of water molecules in type IV (sensu Jeffrey = type III sensu O'Keeffe) clathrate-hydrates, point group P6/mmm, but about two orders of magnitude larger. PMID- 11732065 TI - Autoreproductive cells and plant meristem construction: the case of the tomato cap meristem. AB - Root apical meristems are composed of two zones in which either formative or proliferative cell divisions occur. Within the formative zone, autoreproductive initial cells (a-cells) occupy distinctive locations. By means of graph-L systems, the behavior of one such type of a-cells has been investigated, with particular reference to root caps within the developing primordia of lateral roots of Lycopersicon esculentum cultivated in vitro. Here, the a-cells constitute the "protoderm initials", cells which are found also in the root cap of many angiosperm species. A set of cuboidal (i.e., six-sided) a-cells develops early in the ontogeny of a lateral-root primordium. Then, according to both anatomical observations and theoretical simulations obtained by the application of graph-L-systems, sequential production of descendents from each a-cell leads to the formation of a new autoreproductive cell (a), a cap columella initial (c), and two mother cells (e and f) whose respective descendents differentiate as root epidermis and cap flank cells. In this graph-L-system, there is specification of the location of sister cells with respect to the three orthogonal directions of a cuboidal. In the early stage of root cap formation, only a few rounds of these formative cell divisions by each a-cell and its four types of descendents are required to provide the basic set of cells necessary for full cap development. After the lateral root emerges from the parent root, there may be a temporary cessation of the formative divisions of the a-cells which give rise to columella initials. Columella production is then supported entirely by its own independent set of autoreproductive c-initials. At the same time, division of the autoreproductive protoderm initial cell is directed towards maintaining the cap flank and the epidermal cell files. The regulation of the types of formative division by the a-cell may be represented by means of a division counter which may be specific for a given species. PMID- 11732066 TI - Actin and pollen tube growth. AB - Actin microfilaments (MFs) are essential for the growth of the pollen tube. Although it is well known that MFs, together with myosin, deliver the vesicles required for cell elongation, it is becoming evident that the polymerization of new actin MFs, in a process that is independent of actomyosin-dependent vesicle translocation, is also necessary for cell elongation. Herein we review the recent literature that focuses on this subject, including brief discussions of the actin binding proteins in pollen, and their possible role in regulating actin MF activity. We promote the view that polymerization of new actin MFs polarizes the cytoplasm at the apex of the tube. This process is regulated in part by the apical calcium gradient and by different actin-binding proteins. For example, profilin binds actin monomers and gives the cell control over the initiation of polymerization. A more recently discovered actin-binding protein, villin, stimulates the formation of unipolar bundles of MFs. Villin may also respond to the apical calcium gradient, fragmenting MFs, and thus locally facilitating actin remodeling. While much remains to be discovered, it is nevertheless apparent that actin MFs play a fundamental role in controlling apical cell growth in pollen tubes. PMID- 11732067 TI - Cytoplasmic illuminations: in planta targeting of fluorescent proteins to cellular organelles. AB - Use of the jellyfish green-fluorescent protein as an in vivo reporter is in the process of revolutionising plant cell biology. By fusing the protein to specific targeting peptides or to sequences of complete proteins, it is now possible to observe the location, structure, and dynamics of a number of intracellular organelles over extended periods of time. In this review we discuss the most recent developments and unexpected results originating from the targeting of this unique protein and its derivatives to elements of the cytoskeleton and to membrane-bounded organelles in a range of plant cell types. PMID- 11732069 TI - Questionnaire in clinical research and practice. PMID- 11732070 TI - Clinical features of malaria in Orang Asli population in Pos Piah, Malaysia. AB - A malaria survey was conducted to examine the presence of common clinical features of malaria in individuals living in an endemic area of malaria. The overall infection rate was 11.0% with 7.5% and 3.5% infected with Plasmodium vivax and Plasmodium falciparum respectively. The mean parasitaemia level of both species was 2905.9 parasites/microliter blood, with the mean parasitaemia level of P. vivax and P. falciparum at 682.7 parasites/microliter blood and 6981.7 parasites/microliter blood respectively. The infection rates were higher in the younger age group. Hepatomegaly, hepatosplenomegaly and clinical anaemia were significantly associated with malaria. None of the patients were febrile. In conclusion, in low endemic areas, the presence of clinical anaemia, hepatomegaly and hepatosplenomegaly in afebrile individuals could be considered as useful criteria for the presence of asymptomatic parasitaemia. It is important to carry out laboratory diagnostic investigations, to ensure all the asymptomatic parasitaemia which act as reservoirs are detected and treated. PMID- 11732068 TI - Actin and actin-binding proteins in higher plants. AB - The actin cytoskeleton is a complex and dynamic structure that participates in diverse cellular events which contribute to plant morphogenesis and development. Plant actins and associated actin-binding proteins are encoded by large, differentially expressed gene families. The complexity of these gene families is thought to have been conserved to maintain a pool of protein isovariants with unique properties, thus providing a mechanistic basis for the observed diversity of plant actin functions. Plants contain actin-binding proteins which regulate the supramolecular organization and function of the actin cytoskeleton, including monomer-binding proteins (profilin), severing and dynamizing proteins (ADF/cofilin), and side-binding proteins (fimbrin, 135-ABP/villin, 115-ABP). Although significant progress in documenting the biochemical activities of many of these classes of proteins has been made, the precise roles of actin-binding proteins in vivo awaits clarification by detailed mutational analyses. PMID- 11732071 TI - A study on community acquired pneumonia in adults requiring hospital admission in Penang. AB - A study was carried out to determine the pattern of microbiological organisms causing community acquired pneumonia in adult patients admitted to Penang Hospital between November 1999 and August 2000. Altogether, 98 patients (64 males, 34 females) with a mean age (+/- S.D.) of 55.9 (+/- 19.0) (range 15 to 87) years were included in the study. Causative organisms were identified in 42 patients (42.9%). Mycobacterium tuberculosis was the commonest pathogen being identified in 15.3% of cases, followed by Klebsiella pneumoniae (7.2%), Pseudomonas aeruginosa (6.1%) and Staphylococcus aureus (5.1%). Streptococcus pneumoniae and Acinetobacter spp accounted for 3 cases each (3.1%) and Haemophilus influenzae, non-haemolytic Streptococcus, Mycoplasma pneumoniae, Salmonella typhi, Escherichia coli, Klebsiella spp and Pseudomonas spp for 1 case each (1.0%). Four patients (4.1%) had dual infections and no case of legionella pneumonia was found in this series. PMID- 11732072 TI - Beck Depression Inventory (BDI): a reliability and validity test in the Malaysian urological population. AB - This study aimed to validate the Beck Depression Inventory (BDI) in the Malaysian urological population. Reliability and internal consistency were evaluated using the test-retest method and internal consistency was assessed using Cronbach's alpha. Responsiveness was expressed as the effect size. Internal consistency was high (Cronbach's alpha value = 0.56 to 0.87). Test-retest correlation coefficient and intraclass correlation coefficient were significant (ICC = 0.56 to 0.87) and a high degree of sensitivity and specificity. The BDI is thus a reliable and a valid instrument to be used in Malaysia. PMID- 11732073 TI - A Malaysian Study on the reliability and validity of the Health-Related Quality of Life Questionnaire (HRQOL-20) in urological patients. AB - Main objective of this study is to validate the Health-Related Quality of Life (HRQOL-20) in the Malaysian population. Reliability and internal consistency were evaluated using the test-retest method and Cronbach's alpha. Responsiveness was expressed as the effect size. Internal consistency was excellent (Cronbach's alpha value = 0.68 to 0.87). Test-retest correlation coefficients and intraclass correlation coefficient were significant (ICC = 0.58 and 0.91) as well as the high degree of sensitivity and specificity. The HRQOL-20 is a reliable, valid and sensitive to clinical changes in the Malaysian urological population. PMID- 11732074 TI - Gender differences in the clinical and serological features of systemic lupus erythematosus in Malaysian patients. AB - An analysis of the clinical and serological features of 12 male and 122 female patients with SLE was done to determine whether sex related differences exist. We found a lower incidence of mucocutaneous symptoms and arthritis but an increased incidence of discoid lesions, pleuritis and pericarditis in males at disease onset. During the disease course, there was a lower incidence of arthritis, a similar prevalence of mucocutaneous symptoms but an increased incidence of pleuritis in males with a trend towards renal involvement. These findings were however not statistically significant except for the higher incidence of thrombosis among males. Serologically, both groups showed similar frequencies of autoantibodies and hypocomplementaemia. Although the study was small, it was shown that several sex-related differences in the clinical and serological features exist in Malaysian SLE patients. PMID- 11732075 TI - Mortality patterns in Malaysian systemic lupus erythematosus patients. AB - A retrospective analysis of the case records of 494 systemic lupus erythematosus (SLE) patients under follow-up at University Hospital, Kuala Lumpur during 1976 1990 was performed. Overall mortality was 20.2% (100 patients). The causes of death were infection (30%), renal (15%), respiratory (14%), neurological (5%), cardiovascular (7%), other causes (2%) and unknown (27%). Active SLE was a contributing factor in 19% of the deaths. The patients who died had significantly more renal disease, neurological disease, serositis or thrombocytopenia by the end of the first year of disease compared to the survivors. As in other series, infection and active SLE remain important causes of death. PMID- 11732076 TI - Three-Dimensional Dynamic Subtraction Contrast Enhanced Magnetic Resonance Angiography of the peripheral arteries--an initial experience. AB - This is a prospective study with the objective of comparing Three Dimensional (3D) Dynamic Subtraction Contrast Enhanced Magnetic Resonance Angiography (3DDSCEMRA) with Conventional Catheter Arteriography (CCA) as the gold standard, in the diagnosis of peripheral occlusive disease. Three patients scheduled for CCA in this hospital in between September and October 2000 were included in this study. Patients underwent 3DDSCEMRA before proceeding to CCA on the same day. The 3DDSCEMRA reader was blinded to the CCA results. The results showed good correlation in iliofemoral segment and poor correlation in the infrapopliteal segment due to artifacts in 3DDSCEMRA images. 3DDSCEMRA is a new, non-invasive and promising technique in the diagnosis of peripheral occlusive disease. PMID- 11732077 TI - A study of marketing and its effect on infant feeding practices. AB - A baby milk market was created in the late nineteenth and early twentieth centuries and was conceived through the mutual attraction of the manufacturers and doctors. This has partly attributed to the change from breast to artificial feeding. This study was conducted to determine whether marketing had any effect on infant feeding practices. Fifty Malay mothers whose last child was aged less than five years were asked, by questionnaire and by interview, to identify whether several marketing methods influenced their decision on the selection of infant feeding practice. It was found that these mothers felt milk advertisements on television were attractive (72%), were influenced by others to buy a particular brand of formula (38.4%), were influenced by free milk samples to buy a particular brand of milk formula (65.2%) and were not influenced by their doctors on the chosen method of feeding (68%). More studies should be conducted on a bigger sample in other settings and targeting other methods of marketing to substantiate the above results. Further, similar studies among Chinese and Indian mothers are also required. PMID- 11732078 TI - The timing and stability of choice of medical specialty among Malaysian doctors. AB - A total of 356 doctors responded to a survey on the timing and stability of choice of medical specialty. The majority of doctors made their final decision while working as a medical officer. One hundred (48.5%) of the doctors had made an earlier choice as medical students, 63 (30.6%) during their internship and 43 (20.9%) made their final choice while they were medical officers. Working experience in the specialty was the most important factor in determining final choice of specialty. Advice from consultants/seniors, better financial prospects and parental influence were more important for the male doctors while marriage and family considerations were more important for the female doctors in making their final choice. PMID- 11732079 TI - Use of the Griffiths Mental Scales in normal 2 year old Malaysian children. AB - The Griffiths Scales for Mental Development were used to assess a group of 60 normal 2-year old Malaysian children (25 Indian, 23 Malay and 12 Chinese). The mean GQ was 104.2 (SD 9.3). This was significantly higher than the test mean of 100, p < 0.001. The mean score for Malaysian children was significantly higher on the locomotor, personal social, performance and practical reasoning subscales while they were significantly lower on the hand eye subscale and did not differ from the test mean on the hearing and speech subscale. There was a significant correlation between GQ and social class, r = -0.39, p < 0.05. Scores were lower than those currently obtained on British children, p < 0.001. Minor difficulties due to language and cultural factors arose over the interpretation of several items but with standardisation of these items the test is useful in Malaysian children. PMID- 11732080 TI - Low-dose inhaled nitric oxide in term and near-term infants with hypoxic respiratory failure: a Malaysian experience. AB - Inhaled nitric oxide (iNO) improves oxygenation in term and near-term infants with persistent pulmonary hypertension of the newborn (PPHN) and decreases the need for treatment with extracorporeal membrane oxygenation (ECMO). This mode of treatment is currently being introduced in Malaysia. We report our preliminary experience using low dose inhaled nitric oxide (20 parts per million) in three newborn infants (meconium aspiration syndrome, primary PPHN and congenital diaphragmatic hernia) with severe PPHN who fulfilled criteria for ECMO with a mean oxygenation index (OI) of 40. Two of the infants showed rapid and sustained improvement in oxygenation with a reduction in oxygenation index (OI) over 24 hours. The infant with diaphragmatic hernia showed an initial improvement in OI, which was unsustained and subsequently died. All three infants did not show significant elevation of methemoglobin or nitrogen dioxide (NO2). Inhaled nitric oxide is an effective and safe treatment for severe PPHN that can be used in a developing country like Malaysia. PMID- 11732081 TI - Ocular biometry in the subtypes of primary angle closure glaucoma in University Malaya Medical Centre. AB - Thirty-seven consecutive patients (41 eyes) diagnosed with primary angle closure glaucoma (PACG) attending the Glaucoma Clinic in University Malaya Medical Centre, over a period of 6 months were categorized into acute, subacute and chronic PACG from their clinical presentation. Each case was subjected to automated refraction, A-scan biometry for anterior chamber depth, axial length and lens thickness, keratometry and corneal diameter measurement. Calculations for the relative lens position and the lens thickness: axial length index were performed. The data collected was analysed by the nonparametric test (Kruskal Wallis), one way analysis of variance (ANOVA), chi-square test, Spearman's nonparametric correlations and regression analysis. For controls 15 eyes from 15 normal subjects matched for age, sex, refractive error and race were chosen and subjected to the same examinations. Chronic PACG was the predominant subtype (53.6% of patients and 58.5% of eyes). The ocular biometric measurements of acute PACG eyes deviated most from normals in having the shallowest anterior chamber depth, shortest axial length, smallest corneal diameter, steepest corneal radius, thickest and most anteriorly situated lens, and the greatest lens thickness: axial length index. The subacute subtype was closest to normal and chronic PACG subtype fell in between in most of the biometric characteristics. These findings were not statistically significant. All PACG eyes as a group however showed statistically significant shallower anterior chamber depth (p < 0.05), and a more anterior relative lens position (p < 0.05) compared to normals. PMID- 11732082 TI - Quality of life--how do Malaysian asthmatics fare? AB - Asthma can place considerable restrictions on the physical, emotional and social aspects of the lives of patients. The assessment of quality of life aims to provide a means of measuring the impact of this disease on patients' lives, from the patients' perspective. A cross sectional multi-centre study was conducted in six government hospitals throughout the country. Self-administered SF-36 was used, and clinical information obtained through interviews and examination. 1612 asthmatics responded. Females constituted 63% of the respondents; mean age was 40.9 years; Malays were the majority ethnic group, while 70.8% had secondary level education and 53.7% were employed. Half had suffered from asthma for at least 13 years, while 46.8% and 23.6% have moderate and severe disease respectively. Quality of life was affected by severity of disease. Asthmatics, had a significantly poorer quality of life than the general US population. Severe asthma disease was associated with a compromised quality of life, similar to that of COPD. PMID- 11732083 TI - Lamotrigine as an add-on therapy in intractable paediatric epilepsy--the Kuala Lumpur Hospital experience. AB - An observational study of all children with intractable epilepsy at the Paediatric Institute prescribed Lamotrigine as an add-on therapy between January 1994 and November 1998 was conducted. A total of 30 children were recruited. Three had adverse effects to the drug and it was withdrawn. Of the remaining 27, there were 20 boys and 7 girls, ranging from 2 to 17 years. Fifteen children had generalised epilepsy, 6 had partial epilepsy, 2 had West syndrome and 4 had Lennox Gastaut syndrome. Six children (20%) became seizure free, and 14 (54%) had a greater than 50% reduction in seizure frequency. However 7 children (23%) did not respond and 3 experienced a deterioration in seizure severity. Nine children were noted to have an improvement in alertness and behaviour. Our small series suggests that Lamotrigine is useful as add-on therapy in childhood intractable epilepsy. PMID- 11732084 TI - In vitro activity of sulperazon against recent isolates of ceftazidime-resistant bacteria. AB - The in vitro activity of sulperazon (cefoperazone/sulbactam) was tested against 94 ceftazidime-resistant strains of bacteria isolated from mostly seriously ill patients in critical care units. Acinetobacter baumanii, Pseudomonas aeruginosa and Klebsiella pneumoniae made up 80% of the pathogens studied; 90% of the Klebsiella strains were producers of extended-spectrum beta-lactamases (ESBL). The MIC90 of sulperazon for Klebsiella was 12 mg/l (range 1.5-16 mg/l), indicating that this drug may be a useful alternative for the treatment of ceftazidime-resistant, ESBL-producing Klebsiella. PMID- 11732086 TI - Traumatic transection of aorta. AB - A 16 year-old man presented with fracture of both his femurs after a road traffic accident. Chest radiograph revealed mediastinal widening. Subsequent CT scan and arch aortogram confirmed the findings of traumatic aortic arch transection at the isthmus. He underwent successful surgical repair. High index of suspicion and prompt actions are important in managing this potentially fatal but treatable condition. PMID- 11732085 TI - Balloon endometrial thermoablation--an alternative management of adenomyosis with menorrhagia and dysmenorrhoea. AB - A 45 years old Chinese housewife presented with menorrhagia and dysmenorrhoea due to adenomyosis failed to respond to various medical treatments. She was treated with balloon thermoablation. The total menstrual blood loss (MBL) decreased from 96.94 ml before to 37.57 ml, six months after thermoablation. The pictorial blood loss chart (PBLC) showed similar decrease in blood loss. Dysmenorrhoea was also cured. At three year follow up, there was no recurrence. This is the first report which shows thermoablation decreases MBL objectively and can be tried to treat adenomyosis. PMID- 11732087 TI - Lithium neurotoxicity. AB - Inspite of the advent of newer antimanic drugs, lithium carbonate remains widely used in the treatment and prevention of manic-depressive illness. However care has to be exercised due to its low therapeutic index. The central nervous system and renal system are predominantly affected in acute lithium intoxication and is potentially lethal. The more common side effect involves the central nervous system. It occurs early and is preventable. We describe three cases of lithium toxicity admitted to Johor Bahru Hospital, with emphasis on its neurological preponderance. PMID- 11732088 TI - The use of endoscopic ultrasound and intravenous secretin as an investigative tool to aid diagnosis in pancreatic sphincter hypertension. AB - Pancreatic sphincter hypertension (PSH) is one of the causes of recurrent pancreatitis. The diagnosis can be established by direct measurement of pancreatic sphincter pressures at pancreatic sphincter manometry. This procedure is not without risks, and in cases with PSH, it certainly carries a higher risk of post procedure pancreatitis. The treatment of this disorder is pancreatic sphincterotomy, which on its own carries risk of acute pancreatitis. Therefore it is important to establish the diagnosis reliably before undertaking this procedure. In order to overcome the false positive readings that are possible in sphincter manometry, we proposed to use secretin stimulated endoscopic ultrasound (SSEUS) to measure pancreatic ductal response as an adjunctive method to aid and supplement the diagnosis. Here we describe 3 cases in which this was carried out to optimal effect. PMID- 11732089 TI - Understanding insomnia and its management. PMID- 11732090 TI - Rates of movement and distribution of transposable elements in Drosophila melanogaster: in situ hybridization vs Southern blotting data. AB - Genomic copy numbers and the rates of movement of nine families of transposable elements (TEs) of Drosophila melanogaster were estimated in two sets of mutation accumulation lines: Beltsville and Madrid. Southern blotting was used to screen a large number of samples from both genetic backgrounds for TEs. The Madrid lines were also screened by in situ hybridization of TEs to polytene chromosomes, in order to obtain more detailed information about the behaviour of TEs in the euchromatin. Southern blotting data provided evidence of insertions and excision events in both genetic backgrounds, occurring at rates of approximately 10(-5) and 10(-6) per element copy per generation, respectively. In contrast, in situ data from the Madrid background presented a completely different picture, with no evidence for excisions, and a significantly higher rate of transposition (1.01 x 10(-4)). Direct comparison of the two data sets suggests that the Southern blotting technique had serious deficiencies: (i) it underestimated element abundance; (ii) it revealed less than 30% of the new insertions detected by in situ hybridization; and (iii) changes in the size of restriction fragments from any source were spuriously identified as simultaneous insertion-excision events. Our in situ data are consistent with previous studies, and suggest that selection is the main force controlling element spread by transposition. PMID- 11732091 TI - Dominant modifiers of the polyhomeotic extra-sex-combs phenotype induced by marked P element insertional mutagenesis in Drosophila. AB - Members of the Polycomb group (Pc-G) and trithorax group (trx-G) of genes, as well as the enhancers of trx-G and Pc-G (ETP), function together to maintain segment identity during Drosophila development. In order to obtain new marked P mutations in these genes, we screened for dominant modifiers of the extra-sex combs phenotype displayed by males mutant for the polyhomeotic (ph) gene, a member of the Pc-G group. Five P(lacW) insertions in four different genes were found to stably suppress ph: two are allelic to trithorax, one is the first allele specific to the Minute(2)21C gene, and the remaining two define new trx-G genes, toutatis (tou) in 48A and taranis (tara) in 89B10-13. tou is predicted to encode a 3109 amino acid sequence protein (TOU), which contains a TAM DNA-binding domain, a WAKZ motif, two PHD zinc fingers and a C-terminal bromodomain, and as such is likely to be involved in regulation of chromatin structure as a subunit of a novel chromatin remodelling complex. In a previous study, we found that insertion of a P(ph) transposable element containing ph regulatory sequences creates a high frequency of mutations modifying ph homeotic phenotypes. One such insertion enhanced the ph phenotype and we show that it is a new allele of UbcD1/eff, a gene encoding a ubiquitin-conjugating enzyme that is involved in telomere association and potentially in chromatin remodelling. PMID- 11732092 TI - The speed of Muller's ratchet with background selection, and the degeneration of Y chromosomes. AB - The rate of accumulation of deleterious mutations by Muller's ratchet is investigated in large asexual haploid populations, for a range of parameters with potential biological relevance. The rate of this process is studied by considering a very simple model in which mutations can have two types of effect: either strongly deleterious or mildly deleterious. It is shown that the rate of accumulation of mildly deleterious mutations can be greatly increased by the presence of strongly deleterious mutations, and that this can be predicted from the associated reduction in effective population size (the background selection effect). We also examine the rate of the ratchet when there are two classes of mutation of similar but unequal effects on fitness. The accuracy of analytical approximations for the rate of this process is analysed. Its possible role in causing the degeneration of Y and neo-Y chromosomes is discussed in the light of our present knowledge of deleterious mutation rates and selection coefficients. PMID- 11732093 TI - Difficulties in parentage analysis: the probability that an offspring and parent have the same heterozygous genotype. AB - Parentage studies often estimate the number of parents contributing to half-sib progeny arrays by counting the number of alleles attributed to unshared parents. This approach is compromised when an offspring has the same heterozygous genotype as the shared parent, for then the contribution of the unshared parent cannot be unambiguously deduced. To determine how often such cases occur, formulae for co dominant markers with n alleles are derived here for Ph, the probability that a given heterozygous parent has an offspring with the same heterozygous genotype, and Pa, the probability that a randomly chosen offspring has the same heterozygous genotype as the shared parent. These formulae have been derived assuming Mendelian segregation with either (1) an arbitrary mating system, (2) random mating or (3) mixed mating. The maximum value of Pa under random mating is 0.25 and occurs with any two alleles each at a frequency of 0.5. The behaviour with partial selfing (where reproduction is by selfing with probability s, and random mating otherwise) is more complex. For n < or = 3 alleles, the maximum value of Pa occurs with any two alleles each at a frequency of 0.5 if s < 0.25, and with three equally frequent alleles otherwise. Numerically, the maximum value of Pa for n > or = 4 alleles occurs with n* < or = n alleles at equal frequencies, where the maximizing number of alleles n* is an increasing function of the selfing rate. Analytically, the maximum occurs with all n alleles present and equally frequent if s > or = 2/3. In addition, the potential applicability of these formulae for evolutionary studies is briefly discussed. PMID- 11732094 TI - Multiplicative versus additive selection in relation to genome evolution: a simulation study. AB - The evolution of molecular quantitative traits, such as codon usage bias or base frequencies, can be explained as the result of mutational biases alone, or as the result of mutation and selection. Whereas mutation models can be investigated easily, realistic modelling of selection-directed genome evolution is analytically intractable, and numerical calculations require substantial computer resources. We investigated the evolution of optimal codon frequency under additive and multiplicative effects of selected linked codons. We show that additive selective effects of many linked sites cannot be effective in genomes when the number of selected sites is greater than the effective population size, a realistic assumption according to current molecular data. We then discuss the implications of these results for isochore evolution in vertebrates. PMID- 11732095 TI - Selection and subsequent analysis of sib pair data for QTL detection. AB - Haseman and Elston (1972) developed a robust regression method for the detection of linkage between a marker and a quantitative trait locus (QTL) using sib pair data. The principle underlying this method is that the difference in phenotypes between pairs of sibs becomes larger as they share a decreasing number of alleles at a particular QTL identical by descent (IBD) from their parents. In this case, phenotypically very different sibs will also on average share a proportion of alleles IBD at any marker linked to the QTL that is lower than the expected value of 0.5. Thus, the deviation of the proportion of marker alleles IBD from the expected value in pairs of sibs selected to be phenotypically different (i.e. discordant) can provide a test for the presence of a QTL. A simple regression method for QTL detection in sib pairs selected for high phenotypic differences is presented here. The power of the analytical method was found to be greater than the power obtained using the standard analysis when samples of sib pairs with high phenotypic differences were used. However, the use of discordant sib pairs was found to be less powerful for QTL detection than alternative selective genotyping schemes based on the phenotypic values of the sibs except with intense selection, when its advantage was only marginal. The most effective selection scheme overall was the use of sib pairs from entire families selected on the basis of high within-family variance for the trait in question. There is little effect of selection on QTL position estimates, which are in good agreement with the simulated values. However, QTL variance estimates are biased to a greater or lesser degree, depending on the selection method. PMID- 11732096 TI - A comment on Xie and Xu: 'mapping quantitative trait loci in tetraploid species'. AB - Xie & Xu (2000) present a model for mapping quantitative trait loci in an autotetraploid population. However, one aspect of their model, namely gamete formation, does not properly represent the biological process in autotetraploid species. This paper gives a more realistic formulation for this part of the model, and discusses the consequences for multipoint mapping. PMID- 11732097 TI - [Indications and contraindications for immediate hospitalization]. PMID- 11732098 TI - [Spectrum of diagnoses in an internal medicine emergency unit]. PMID- 11732099 TI - [Urological emergencies in house calls by internists]. PMID- 11732100 TI - [Gynecologic emergencies]. PMID- 11732101 TI - [Functional (somatoform) disorders in house calls by internists]. PMID- 11732102 TI - [Legal aspects of house calls]. PMID- 11732103 TI - [Chronic fatigue]. PMID- 11732104 TI - [Recurrent hemoptysis in a 32-year old female patient with complications stemming from surgery for aortic isthmus stenosis in childhood]. PMID- 11732105 TI - [Twenty-seven year old female patient with sensation of coldness and burning pain in the left hand]. PMID- 11732107 TI - [Benefits and limitations of intensive care. Report from the 33. Annual Meeting of the German Association for Internal Medicine Intensive Care and Emergency Medicine, June 20-23, 2001 in Halle/Saale ]. PMID- 11732106 TI - [Insulin analogs: structure, properties and therapeutic indications. Part 1: rapid-acting insulin analogs]. PMID- 11732110 TI - [Agglutination phenomena of normal human blood]. PMID- 11732111 TI - [Karl Landsteiner]. AB - Karl Landsteiner received the Nobel Prize for Medicine in 1930. He worked in Vienna, The Hague and New York, making fundamental contributions in the field of immune hematology (blood groups and Rhesus factor), syphilis (visualising spirochetes in the dark field) and poliomyelitis (viral genesis). In particular, his findings in the general field of serology and immunology (haptens, specificity of antigens) have not lost any of their significance and are valid to this day. PMID- 11732112 TI - [Etiological research in medicine in Vienna circa 1900, at the time of Karl Landsteiner]. AB - By the end of the 19th century theoretical etiological research became more and more important in medical science. Anton Weichselbaum focused on bacteriology in the field of pathological anatomy and Rudolf Paltauf founded an Institute of Serotherapy, thus taking account of this new development. Progress made in laboratory medicine due to the work of a number of scientists in Vienna was of both fundamental and practical significance for the advancement of medicine. PMID- 11732113 TI - A century of human blood groups. AB - During the course of the twentieth century, at least 270 authenticated alloantigens have been recognised on the red cell surface. Most of these have been classified into 26 blood group systems, each of which represents a single gene or a cluster of two or three closely-linked homologous genes. Most blood group polymorphisms result from single nucleotide changes encoding amino acid substitutions in cell surface proteins. Many other mechanisms, including recombination between homologous genes, are also involved, especially in the complex Rh and MNS systems. There are at least three common molecular backgrounds to the RhD-negative phenotype. Some blood group antigens are carbohydrates, the polymorphisms resulting from mutations within genes encoding glycosyltransferases. Red cell surface proteins perform a variety of functions. For some the functions are well understood, but for most they can only be surmised from the structure of the protein. Putative functions include, membrane transport, cell adhesion, complement inactivation, binding chemokines, and anchoring the plasma membrane to the cytoskeleton. Some erythroid cell surface antigens may serve their primary purpose during erythropoiesis. Analysis of the development of these proteins on erythroid cells during erythropoiesis, ex vivo, has provided clues to their functions and a useful set of markers for the study of erythropoiesis. PMID- 11732114 TI - [Biochemical basis of the ABO system subgroups]. AB - This review article offers a survey of the biochemical and molecular biological basis of the ABO subgroups. In contrast to protein-based blood groups, the antigens of the ABO system are carbohydrate antigens. The determinant carbohydrate structures are synthesised stepwise by the action of glycosyltransferases which transfer single monosaccharide residues onto an appropriate precursor substance. The genes responsible for the formation of the antigens of the ABO system (viz. ABO, Hh, and Sese) each encode single glycosyltransferases. Sequence analyses have shown that mutations in the ABO gene affecting only two amino acids in the encoded glycosyltransferase decide whether A or B epitope is synthesised. In blood group O individuals several inactive alleles of the ABO locus have been detected, which encode inactive transferases. These mutilated enzymes are not able to synthesise a defined blood group specificity. Blood group O is therefore characterised by the presence of unchanged A and B precursor, i.e. H-substance. Similarly, inactive variants of the H gene are responsible for the rare 'Bombay' phenotype and inactive variants of the secretor gene (Se) are responsible for the non-secretor status. In both cases the mutant transferases are not able to synthesise H-substances. PMID- 11732115 TI - Granulocyte immunology. AB - Antibodies directed against antigens on the granulocyte (neutrophil) membrane can cause a variety of disorders including neonatal immune neutropenia, immune neutropenia after bone marrow transplantation, autoimmune neutropenia, and drug induced immune neutropenia. Since granulocyte alloantibodies can lead to severe pulmonary transfusion reactions (TRALI), febrile transfusion reactions and refractoriness to granulocyte transfusions, they also play an important part in blood transfusion. The implicated human neutrophil alloantigens (HNA) have been renamed in the recently introduced HNA nomenclature which is based on the antigen's glycoprotein location. The Fc gamma Receptor IIIb (CD16, HNA-1) and the NB1 glycoprotein (CD177, HNA-2) represent the major immunogenic molecules of the neutrophil membrane. They bear the clinically most important antigens HNA-1a,-1b, 1c (NA1, NA2, SH) and HNA-2a (NB1), respectively. For the detection of granulocyte antibodies, a combination of immunofluorescence and agglutination tests together with a panel of freshly isolated, typed test neutrophils has been shown to represent the best means of detection. The introduction of the glycoprotein-specific assay "MAIGA" has improved alloantibody identification considerably. To facilitate and improve neutrophil typing, PCR-SSP techniques have been established for HNA-1a,-1b, and -1c genotyping. PMID- 11732116 TI - Human platelet alloantigens. AB - Antibody formation against alloantigens of the human platelet membrane is responsible for clinical syndromes and transfusion related conditions as neonatal alloimmune thrombocytopenia (NAIT), post-transfusion purpura (PTP), platelet transfusion refractoriness (PTR) and passive alloimmune thrombocytopenia. Moreover, rare cases of alloimmune reactions involving platelets have been observed after transplantation of hematopoietic stem cells. Among alloantigens of the platelet membrane shared with other cells (type I alloantigens) are the glycoconjugates of the ABO system and class I human leukocyte antigen (HLA) antigens. Antibodies against these structures are responsible for PTR and for febrile nonhemolytic transfusion reactions. Antibodies against type II antigens (formerly termed "platelet specific antigens") have been observed in NAIT, PTP and passive alloimmune thrombocytopenia. ABH antigens have been identified on intrinsic platelet membrane glycoproteins. Moreover, it is now clear that HLA class I antigens are an integral part of the platelet membrane. The quantity of both HLA and ABH-antigen expression on the platelet membrane varies considerably. Single point mutations account for almost all platelet specific alloantigens, but most antigenic determinants seem to depend upon glycoprotein conformation: generally, platelet specific alloantibodies fail to recognize synthetic peptides encompassing the polymorphic residues. Restriction fragment polymorphism analysis and allele-specific PCR have been implemented for genotyping of platelet alloantigens in many laboratories. Antigen specific assays using monoclonal antibodies (MAIPA, immunobead assay) became de facto standard for diagnosis of platelet antibodies in serum/plasma samples. It can be expected that innovative techniques as human alloantibody fragments produced by phage display technique and the production of recombinant antigens will allow rapid and reliable phenotyping and antibody detection in the future. PMID- 11732117 TI - Molecular genetics of the HLA complex. AB - The Human Leukocyte Antigen (HLA) system can be defined as a set of glycoproteins which take up peptides intracellulary and become ligands for immune receptors once they are expressed on the cell membrane. Functional HLA molecules are trimers consisting of a heavy chain and a light chain which bind a peptide. The production of HLA molecules in the endoplasmic reticulum, their assembly and their trafficking to the cell membrane have been studied in great detail. A salient feature of HLA molecules is their polymorphism which is essentially a tremendous amount of variation in the amino acid sequence of mainly the heavy chain between different HLA types. Polymorphic chains of HLA molecules are encoded in the human major histocompatibility complex (MHC) on the short arm of chromosome 6. A map of the nucleotide sequence of the human MHC has been established. The polymorphism of HLA molecules, i.e. the HLA type, is intimately associated with the nature of the peptides bound. In principle, HLA molecules can bind a variety of different peptides. However, peptides with distinct biochemical features are preferentially bound according to the HLA-type. HLA-typing is applied clinically in transplantation and disease association. Recently, recombinant HLA molecules have been used as a tool to define the T cell receptor repertoire. PMID- 11732118 TI - IgG-Fc receptors and the clinical relevance of their polymorphisms. AB - Receptors for the Fc part of IgG form the bridge between immune complexes, antibodies and blood cells. Besides phagocytes, such as granulocytes and monocytes, also lymphocytes and platelets express members of the large family of IgG-Fc receptors or Fc gamma Rs. Three main classes of leukocyte Fc gamma Rs can be distinguished: Fc gamma RI, Fc gamma RII and Fc gamma RIII. Depending on the type of intracellular domain of the leukocyte Fc gamma Rs, interaction of a cell with an antibody-sensitized particle will induce either an activating signal (leading to phagocytosis, degranulation or toxic oxygen formation) or, in case of Fc gamma RIIb, an inhibiting signal. The inhibiting role of Fc gamma RIIb has long been thought to be only important for B-cell responses, however, recently it has been indicated that Fc gamma RIIb may have a much broader role in immune response, also regulating responses of phagocytes. In general, leukocyte Fc gamma Rs have high affinity for IgG1 and IgG3 subclasses. However, the affinity for the various IgG subclasses is influenced by receptor polymorphisms encoded by single nucleotide polymorphisms of the Fc gamma RIIa and Fc gamma RIIIa and Fc gamma RIIIb encoding genes. Because these subtle Fc gamma R changes determine the level of clearance of immune complexes, associations between Fc gamma R isoforms and disease risks have been investigated and also indicated as discussed in this review. Next to the leukocyte Fc gamma Rs, an IgG transport receptor, FcRn (neonatal Fc receptor) has been described. This receptor seems to be important for placental IgG transport and for IgG plasma level homeostasis. FcRn is expressed by endothelial cells and hepatocytes and functions in these cells as a receptor rescuing IgG from destruction, thereby increasing IgG half life. Also blood cells express FcRn and the exact role of FcRn in these cells is still to determined. Thus, although a lot is known, questions on the exact pattern of expression of the IgG receptors and their role in immune response still remain. PMID- 11732119 TI - Rejection mechanisms in transplantation. AB - While major improvements have been made in the prevention and treatment of hyperacute and acute transplant rejection, most grafts will succumb to chronic rejection: this reflects the extent of our knowledge of the mechanisms that drive these processes. Clinically, transplant rejection is classified according to timeframe and histology into hyperacute (minutes to hours), acute (days to months) and chronic rejection (months to years). Hyperacute and acute rejection are reasonably well understood and occur by immune mediated events whereas chronic rejection probably has immune and non-immune components. The trigger to cell-mediated rejection is allorecognition, where same-species, non-self antigens are detected by the host immune system. This occurs by two distinct mechanisms, called the direct and indirect pathways. The direct pathway results from the recognition of foreign major histocompatibility molecules, intact, on the surface of donor cells. Indirect allorecognition occurs when donor histocompatibility molecules are internalised, processed, and presented as peptides by host antigen presenting cells. Animal and human studies strongly suggest that acute rejection is predominantly triggered by the direct pathway although if the latter is blocked then the indirect pathway can suffice. Donor antigen presenting cells within the graft become depleted with time and the frequency of T cells reactive to the direct pathway diminishes irrespective of whether or not chronic rejection occurs. This implies that the direct pathway is unlikely to contribute to chronic rejection. Assays of T cell responses have, however, found an association between the indirect pathway and chronic rejection although it is unlikely that this is the whole story: there are numerous non-immunological risk factors for chronic rejection which probably interact with immune components causing gradual graft failure. Xenotransplantation, where tissue is transferred across species, causes rejection by processes analogous to those seen in allografts but they are faster and more vigorous. Novel approaches have overcome some early antibody mediated rejection events but then reveal a huge, intense, adaptive cellular response. We believe that by the careful study of the mechanisms of rejection, the problems of chronic rejection and xenograft rejection will be overcome, thus reversing the widening gap between organ demand and supply. PMID- 11732120 TI - [Worldwide eradication of poliomyelitis]. AB - Poliomyelitis, an infectious disease with acute and persistent flaccid paralysis is caused by poliovirus (types 1, 2 or 3), an enterovirus. The infection is asymptomatic in 95% of infected subjects. Most of the paralytic cases occur in adolescents or adults in the course of polio type 1 infection. In the prevaccination era, in countries with poor hygienic conditions, infection in early childhood was common, mostly asymptomatic, and immunity in the population prevailed. In developed countries polio often struck adolescents and adults taking its toll in paralytic disease. The introduction of vaccination with the Salk vaccine (IPV Inactivated Polio Vaccine) in the USA and in Europe in 1956 and with the Oral Polio Vaccine (OPV) developed by Sabin worldwide in the early sixties made it possible to control the epidemic in large geographic areas, but it could not eliminate the disease worldwide. Poliomyelitis is still endemic in Central Africa and in the Indian sub-continent. Acts of war led to the reduction in the vaccination rate in different geographic areas, and smaller epidemics with wild virus but also with reverted vaccine strains occurred. In some parts of the world the rate of vaccination also declined due to elimination of poliomyelitis, and it came to small epidemics of paralytic polio mainly caused by reverted vaccine strains circulating in the population. Reverted vaccine strains also remain a central problem in the eradication of poliomyelitis projected for 2005 by the World Health Organisation. A high vaccination rate, preferably with 3 doses of OPV in infancy or early childhood, and exact worldwide monitoring of cases is indispensable for the eradication. For the complete eradication of poliovirus the live vaccine OPV would have to be changed to an inactivated vaccine IPV worldwide. However, this is presently unachieveable, because of logistic problems and high costs. PMID- 11732121 TI - The pediatric genitourinary examination. Inguinal, urethral, and genital diseases. AB - This article covers a wide spectrum of genitourinary pathology. It is hoped that it will provide a point of reference for the genitourinary examination, guidance for effective referral, and a reference for initial parental counseling. PMID- 11732122 TI - Genitourinary imaging in children. AB - Genitourinary imaging in children has gone through a metamorphosis in the past 30 years, paralleling technical advances in sonography, CT scanning, and MR imaging. The imaging sequences for hydronephrosis and UTI are still controversial and require outcomes analysis to determine which studies are most beneficial and efficacious. PMID- 11732124 TI - Pediatric urinary tract infections. AB - Urinary tract infection in the pediatric population can lead to significant morbidity if not treated promptly and appropriately. All first infections may signify possible underlying anatomic or functional abnormality and require imaging of the lower and upper tracts. Accurate diagnosis of UTIs requires a properly collected quantitative urine culture. Treatment should be tailored to the pathogen as dictated by the urine culture sensitivities to minimize the development of multidrug-resistant organisms. Prophylactic agents should differ from the antibiotic used in the acute setting and preferably concentrated in the urinary tract, with minimal effects on the normal fecal flora. In the long term, patients with documented evidence of renal scarring should be followed up for signs of renal deterioration and hypertension. PMID- 11732123 TI - Pediatric urine testing. AB - Today, urinalysis is one of the most common clinical tests ordered for adult and pediatric patients. Because urine specimens are usually readily available and are obtained noninvasively, the urine testing is well suited for children. This article discusses the most common urine tests performed in children for screening purposes and also less common tests for diagnosis of specific disorders. Special considerations regarding urine specimen collection in children are discussed. Some simple tests that are underused by clinicians are mentioned, as are some exciting new molecular applications of urine testing. PMID- 11732125 TI - Enuresis. AB - The authors do not have all of the data about enuresis, and many children are subject to relapses or failure of treatment. There is no cause for despondency, however. Enuresis is no longer a mystery. Good data exist about the natural history, epidemiology, and etiology of enuresis. In addition, multiple treatment modalities are available to practitioners. This article has sought to review the scientific literature and to relate the authors' experience with enuresis. The authors recommend a treatment program for children with monosymptomatic nocturnal enuresis that includes removal of caffeine from the diet. Enuretic children do not consume enough fluid, and the authors recommend that the daily fluid requirement be divided during the day: 40% in the morning, 40% in the afternoon, and 20% in the evening, with no restriction of fluid. Normalization of small functional bladder capacities may help to cure enuresis and has an effect on the efficacy of other therapies. Treatment of enuretics with antibiotics is effective in children with UTI, bacteriuria, or cystitis cystica. DDAVP has been shown to be effective in the treatment of enuresis, especially in children who have achieved a normal functional bladder capacity. Bladder alarm systems also offer a potential cure of enuresis, are inexpensive, and show a low relapse rate. PMID- 11732126 TI - Dysfunctional elimination syndrome in children without obvious spinal cord diseases. AB - Elimination dysfunction is a functional disease that encompasses the urinary and GI tracts. Because the presentation is varied and ED is a disease of exclusion, a thorough history and physical examination are mandatory. Noninvasive management that includes the "fundamental four" and continuous engagement and encouragement of the patient and family are the pillars of treatment. For patients who are refractory to this approach, new research in children-directed biofeedback research shows promise. PMID- 11732127 TI - Ureteropelvic junction obstruction in infants and children. AB - An important question that needs to be answered in larger numbers of children is the potential recoverability of function in those followed up until function decreases. Most series involve small numbers. Some investigators claim that the function, once lost, does not return; others believe that early repair yields better improvement than does repair after 1 year. In the meantime, the authors continue to recommend a restrained interventional approach. The authors use surgical therapy early for infants with grade 3 or 4 dilatation and decreased function in the involved kidney or, overall, those with solitary kidneys or bilateral involvement. Those followed up undergo surgery if they show increased dilatation or loss of function, develop symptoms, or have dilatation that does not improve beyond the second year of life. The authors prefer to operate before there is loss of function, even at the risk of operating in a few infants that might have remained stable, because the long-term outcome of those who remain dilated is unknown and follow-up in some environments is not always reliable. Whether improved analysis of the excretory pattern on renography will prove to have prognostic value is unknown. PMID- 11732128 TI - Hematuria in children. AB - Children with hematuria require a thorough history and physical examination. Not all children with hematuria require the same investigations. The only laboratory test uniformly required for children with the various presentations of hematuria is a complete urinalysis with a microscopic examination. The rest of the evaluation is tailored according to the pertinent history, physical examination, and other abnormalities on the urinalysis. This article has provided a brief summary of the more common causes of pediatric hematuria and suggestions for tailoring the patient's evaluation according to the presentation. Most causes of hematuria in pediatrics represent medical conditions that often require referral to a pediatric nephrologist. Indications for referral to a urologist are more limited and include stones that do not pass spontaneously or are more than 5 mm in diameter, renal injury from trauma, anatomic abnormalities, or gross hematuria that seems to originate from the urinary tract and is without an identified cause. PMID- 11732129 TI - Neonatal circumcision. AB - The merits of neonatal circumcision continue to be debated hotly. Some argue that circumcision is a "uniquely American medical enigma." Most of the world's male population remains uncircumcised; however, most boys born in the United States continue to undergo neonatal circumcision. Review of existing literature supports that most children who are uncircumcised do well from a medical standpoint and, thus, the question of whether US health care practitioners are subjecting neonates to an unnecessary surgical procedure remains. The medical benefits of circumcision are multiple, but most are small. The clearest medical benefit of circumcision is the relative reduction in the risk for a UTI, especially in early infancy. Although this risk [figure: see text] is real, the absolute numbers are small (risk ranges from 1 in 100 to 1 in 1000), and one investigator has estimated that it may take approximately 80 neonatal circumcisions to prevent one UTI. In the case of a patient with known urologic abnormalities that predispose to UTI, neonatal circumcision has a clearer role in terms of medical benefit to the patient. Most of the other medical benefits of circumcision probably can be realized without circumcision as long as access to clean water and proper penile hygiene are achieved. Proper penile hygiene should all but eliminate the risk for foreskin-related medical problems that will require circumcision. Moreover, proper hygiene and access to clean water has been shown to reduce the rate of development of squamous cell carcinoma of the penis in the uncircumcised population. Proper techniques on the care of the foreskin are illustrated in the American Academy of Pediatrics pamphlet titled "How to care for the uncircumcised penis." Regarding the relationship between STDs and circumcision, patient education and the practice of low-risk sexual behavior make a far greater impact than does routine circumcision in hopes of reducing the spread of HIV and other STDs. Nevertheless, in areas where safe sexual practices are poorly adhered to, circumcision can have a relative protective effect against the transmission of HIV and other STDs. The medical harms of circumcision lie mainly in the 1% acute complication rate and the additional patients who require revision of their initial circumcision for cosmetic or medical reasons. Anecdotally, the authors see far fewer complications in the acute and long-term phase when the circumcision has been performed by someone with expertise and experience with the procedure. Thus, the authors routinely recommend to parents that, if they choose to have their newborns circumcised, they should seek out an experienced practitioner. A negative psychologic and sexual impact of circumcision has been argued, but solid, scientific data are lacking. Special interest groups have argued that perhaps the greatest harm of circumcision is in performing an operation without a clear indication. Many of these groups have claimed that performing a routine neonatal circumcision is akin to performing a surgical procedure without a clear medical benefit, and in an infant, that is akin to surgery without informed consent. Although this is an extreme posture, the clinician can understand the emphasis on trying to provide invasive medical services only when a clear medical benefit is expected, especially when treating an infant or child. Deciding whether or not to circumcise an infant continues to challenge many new parents. Clearly, the procedure provides potential medical benefits and potential risks. It is difficult to say whether the benefits outweigh the risks for all male infants. Further complicating the decision for many American parents is that, in some areas of the United States, there exists an unexplained positive cultural connotation with neonatal circumcision. For these reasons, parents who actively choose to keep their sons uncircumcised need to be encouraged to make this decision forthrightly. Parents who choose to have their children circumcised also should be encouraged to actively seek an experienced practitioner who can afford the child adequate local analgesia. PMID- 11732130 TI - Adolescent varicocele. AB - The evaluation and management of adolescents with varicoceles continue to evolve. Current recommendations for repair are based on the findings of impaired testicular growth or spermatogenesis; however, with early evaluation and selective treatment, clinicians should be able to reduce the potential for future fertility problems significantly in adolescents with varicoceles. PMID- 11732131 TI - Pediatric urolithiasis. AB - Pediatric stone disease is a frequently underestimated entity that can present unique problems in its management. The condition stems from any of a number of underlying, causative factors, and the significant possibility of recurrence must always be borne in mind. The primary care physician should be aware that the condition is readily treatable by an experienced pediatric urologist who has a range of treatment modalities at his or her disposal; however, overall success is governed by a timely referral and subsequent long-term follow-up to ensure the maintenance of a stone-free state. PMID- 11732132 TI - Dysfunction of the lower urinary and distal gastrointestinal tracts in pediatric patients with known spinal cord problems. AB - Destruction of the urinary tract in children with elimination, storage, and holding dysfunction of the lower urinary and the distal GI tracts is caused primarily by high intravesical pressure. UTI accelerates this process. The LPP and the status of the urethral control mechanism and its relationship to the detrusor are the primary determinants of intravesical pressure. Intravesical pressures of more than 40 cm H2O are dangerous because they cause a pressure gradient that is transmitted proximally to the renal papillae, which results in the cessation of renal blood flow and a loss of renal function over time. Hydroureteronephrosis, VUR, UTI, urinary incontinence, and calculi formation also may occur. If these dangerously high intravesical pressures remain untreated, renal failure is likely to occur over time. These children then require dialysis or renal transplantation to survive, which is tragic and represents an enormous economic cost to society. Renal failure and upper urinary tract damage is nearly 100% preventable with early and appropriate evaluation and treatment. CIC is a crucial part of the management of these children and has been shown to be safe and effective, even in newborn boys. The use of the Crede maneuver (i.e., manual compression) to empty the bladder is obsolete and should be abandoned. The distal GI tract is inseparable from the lower urinary tract and must be treated simultaneously. Failure to treat the distal GI tract yields poor clinical results and much patient dissatisfaction and makes it difficult or impossible to treat the child's urinary tract problem successfully. Bowel-management programs must include daily high water and fiber intake, together with digital perianal stimulation or fecal extraction. Neuropathic bladder and bowel problems that are intractable to conservative medical and mechanical (i.e., CIC and digital perianal stimulation or fecal extraction, respectively) management almost always can be corrected surgically with high success rates in cooperative patients. Finally, neuropathic bladder and bowel problems can be extremely isolating and debilitating problems. Psychologic counseling and emotional support must be provided as needed. The care that these patients receive must be organized, comprehensive, and correlated with these patients' lifestyles. If these children are evaluated and treated early, they have the potential to live long, healthy, and productive lives. PMID- 11732133 TI - [Dental--mental]. PMID- 11732134 TI - [Evaluation of patient's level of functioning in the dental practice]. AB - In order to improve the reliability of diagnostic judgements of complex or persisting dental complaints it is important to be aware of the role of emotional problems regarding the origin and the maintenance of dental complaints. In this paper the focus is on the collection of information about patients' history through an anamnesis, and the observation of patients behaviour and mood. PMID- 11732135 TI - [Patients with mental problems]. AB - Epidemiological studies show that one out of four Dutch individuals suffers from a mental disorder. Consequently, dentists are confronted with noticeable deviant or problematic behaviour. In this paper an overview is presented of several of psychological problems and their relevance to dental treatment. This is illustrated by a case description. For a proper diagnosis and adequate treatment planning it is important that dentists have some general knowledge of psychological functioning of patients, and the possible psychological determinants of illness and well-being. This would prevent unnecessary damage of dental tissues and needless suffering but also frustration, both for the patient and the dentist. PMID- 11732136 TI - [Dental complaints are found always between the ears. Somatization in the dental practice]. AB - The aim of the present study was to investigate the occurrence of somatization specific behaviour in the dental setting and its relationship with patients' report of both dental and psychological complaints. Somatization-specific behaviour was operationalized as an unexplained dental complaint, high dental attendance, a high treatment consumption or an unreasonable demand with regard to treatment. Of the 309 patients 8.7% fulfilled the criteria for somatization specific behaviour, which was mainly expressed as a high frequency of attendance (6.8%). Women showed significantly more often (73%) somatization-specific behaviour than men (27%). Further, a relation between depression and somatization specific behaviour was found. Particularly patients with a relatively high level of long-lasting dental complaints demonstrated somatization-specific behaviour. PMID- 11732137 TI - [Crying for attention: patients who mimic manifestations of disease]. AB - This article focuses on the different forms of factitious diseases. Patients may mimic or produce the manifestations of a disease in order to attract attention of doctors. Sometimes patients inflict such serious injuries on their own bodies that they need to undergo long-term treatment in a hospital such as in the case of Munchausen syndrome. Recognition of alarm signals associated with this type of condition may help to avoid a long lasting process of illness, screening and unnecessary treatments. PMID- 11732138 TI - [Satisfied patients?]. AB - This paper examines the difficult question how satisfied patients are with dental services. Scientific research shows that this is hard to measure. Five issues affect dental satisfaction, of which interpersonal and psychosocial factors are reported as being among the most important factors. Two case reports of dissatisfied patients are presented. It is emphasized that adequate counseling skills of the dentist, his evaluation of the expectations and wishes of the patient at the beginning of the treatment, and a sound dental, medical and psychosocial anamnesis are crucial for preventing dental dissatisfaction among patients. PMID- 11732139 TI - [Burnout among dentists: a question of coping]. AB - Work stress and burnout are considered to be serious professional risks in dentistry. This study investigated the relation between a dentist's coping style and burnout level. It was hypothesized that dentists characterized by an active coping style would show relatively low burnout levels, whereas dentists characterized by a passive coping style would show relatively high burnout levels. From a survey among 709 active general dental practitioners, who filled out the Maslach Burnout Inventory and the Utrecht Coping List, this hypothesis was supported. It was concluded that coping style could explain differences in burnout levels to a substantial degree. In the discussion the need for feedback on one's coping style, and the importance of an individual dentist's active attitude towards career planning, among other things, are emphasized. PMID- 11732140 TI - [Extremely anxious patients: does treatment achieve the desired effect?]. AB - The aim of the present study was to determine the outcome of treatment of several treatment modalities utilized at the Centre for Special Dental Care in Amsterdam, the Netherlands. Outcome was assessed in 341 patients in terms of dental anxiety after treatment (response 65%) and dental attendance one year later (response 74%). After treatment, patients treated with a behavioral management approach had lower dental anxiety scores than patients treated with intravenous sedation. Furthermore, one year after treatment 66.7% of the treated patients indicated to visit a general dental practitioner regularly. It is concluded that treatment was effective. However, it was felt that dental attendance level could be increased by a change in the referral procedure to general dental practitioners. It is recommended that a psychologist is involved in diagnosis and treatment planning in dental fear clinics. PMID- 11732142 TI - [Dental care of mentally retarded persons]. AB - To let the dental treatment of mentally handicapped people run as smoothly as possible a good cooperation between dentist and parents, or attendants, is of great importance. After the psychosocial, the medical and the dental anamneses a provisional treatment plan can be made in consultation with the patient and/or his attendants. The treatment plan and the treatment itself are regularly evaluated and, if necessary, adjusted. The cooperation, the nature of the handicap, the oral care and motivation play an important role in determining the aim of the treatment. In most cases an approach directed towards a change in behaviour will suffice. Sometimes, however, pharmacological support is required. For this the patient will usually be referred to a centre for special care dentistry. PMID- 11732141 TI - [Dental fear in children: dental experiences during childhood]. AB - This study aimed to examine the relative importance of invasive treatment experiences in the acquisition of dental fear in children. For this purpose, the complete dental history of 401 children (5-10 years) from 2 dental practices was studied. The level of dental fear in these children was assessed using the Dental Subscale of the Children's Fear Survey Schedule (CFSS-DS). Differences in treatment variables between high fearful and low fearful children were analysed, and regression analysis was performed to determine significant predictors of dental fear. A significant though weak relation of dental fear with the number of experienced extractions was found, while no relation with the number of experienced fillings was found. The results indicate that within the (direct) conditioning pathway, objective dental procedures seem to play a minor role in children's fear acquisition. Clinical support for the 'latent inhibition' theory was provided: a history of neutral or positive dental visits seems to serve as a defence against the development of dental fear in children. PMID- 11732143 TI - [Anxiety disorders]. PMID- 11732145 TI - Congenital heart disease in Papua New Guinean children. AB - The aim of the study was to analyse critically the programme for surgical management of children in Papua New Guinea (PNG) with congenital heart disease. A hospital record-based analysis was undertaken to document the pattern, management and short-term outcome of surgery in PNG children referred with a diagnosis of congenital heart disease to the Royal Alexandra Hospital for Children in Sydney, Australia. On admission, physical examination, chest radiogram, electrocardiogram, cross-sectional echocardiogram and, in most cases, cardiac catheterization were performed. Of the 170 children referred over the 17-year period, 1978-1994, 165 were confirmed to have congenital heart disease and were included in the study. Their ages ranged from 2 months to 16 years (median 5.5) and the male to female ratio was 1:1. One-sixth had delayed milestones and one fifth long-term wasting. A large number were tachypnoeic, in heart failure or had pulmonary hypertension on admission. Ventricular septal defect, 34%, tetralogy of Fallot, 23%, and patent ductus arteriosus, 16.4%, were the predominant defects. lesions such as aortic stenosis, coarctation of the aorta and transposition of the great arteries are under-represented. Altogether, 133 children (81%) had surgery; 75% were open- and 25% closed-heart operations. The complications were unremarkable and the mortality rate (6%) acceptable for the era. The programme was therefore very successful for a small proportion of children born in PNG with congenital heart disease. PMID- 11732146 TI - Respiratory rate as a predictor of positive lung aspirates in young Gambian children with lobar pneumonia. AB - Clinical predictors of a positive bacterial culture from lung aspirate or blood culture were investigated in 90 children under 5 years of age with lobar pneumonia on whom both lung aspiration and blood culture were performed. Of the 66 children with a respiratory rate of > or = 50 breaths/min, 35 (53%) had positive bacterial lung aspirates compared with only five (21.7%) of 23 children with a respiratory rate of < 50 breaths/min (odds ratio [OR] 4.06, 95% confidence interval [CI] 1.24-15.46, p = 0.02). Of the 41 children with positive lung aspirates, 31 (76%) had negative blood cultures. In contrast with children with positive lung aspirates, there were no clinical predictors of a positive blood culture. A respiratory rate of > or = 50 breaths/min in children with radiological evidence of lobar pneumonia would support lung aspiration as a positive result is significantly more likely than in children with a lower respiratory rate. PMID- 11732147 TI - The effect of adjuvant steroid treatment on serial cerebrospinal fluid changes in tuberculous meningitis. AB - Three recent studies found that corticosteroids improve clinical outcome and mortality in tuberculous meningitis (TBM), although the exact mechanism of action of the drug remains speculative. A number of reports on the effect of corticosteroids on cerebrospinal fluid (CSF) findings in TBM have been published, often with conflicting results regarding serial cell counts and protein levels. As part of a controlled, randomized trial on the effect of oral prednisone on outcome in childhood TBM at our institution, CSF was collected and analysed weekly during the 1st month of treatment. We found no significant difference in serial CSF cell counts between the steroid and non-steroid groups in the study. However, the steroid group had significantly lower CSF protein and globulin levels after the 1st month of treatment, and a more steady rise in CSF glucose levels than the non-steroid group. Knowledge of the different CSF responses during the course of anti-tuberculosis therapy is important in clinical decision making. PMID- 11732148 TI - Treatment of falciparum malaria in Vietnamese children: the need for combination therapy and optimized dosage regimens. AB - To assess the in vivo sensitivity of Plasmodium falciparum to mefloquine and artesunate in a hyperendemic area of southern Viet Nam, we studied 41 children and 21 adults from a remote commune who had uncomplicated falciparum malaria without previous treatment. Patients were randomly allocated to artesunate (4 mg/kg on day 0 and 2 mg/kg on days 1-4) or mefloquine (10 mg/kg followed by 5 mg/kg at 6 h). Serial assessments were performed over 28 days. Of 31 patients allocated artesunate, nine (29%) redeveloped parasitaemia during follow-up compared with 23% (seven of 30) who received mefloquine. Of the 41 children, 15 (37%) had recrudescence/re-infection compared with only one of 20 adults (5%; p < 0.001). Significantly more children than adults failed on mefloquine treatment (37% vs 0%; p = 0.021) and one case showed RIII resistance. There was no significant difference in the case of artesunate. In regression analysis, parasitaemia was an independent predictor of recrudescence/re-infection after mefloquine (p = 0.02). These data support the use of combination therapy such as artesunate plus mefloquine for falciparum malaria in a hyperendemic area of Viet Nam. Primarily because of their greater parasite densities, children should be given higher doses of mefloquine (e.g. 25 mg/kg). PMID- 11732149 TI - Seven years' experience with Cryptosporidium parvum in Guinea-Bissau, West Africa. AB - In community-based studies conducted from 1991 to 1997 in Guinea-Bissau, West Africa, stool specimens from children aged less than 5 years with diarrhoea were routinely examined for enteric parasites. Cryptosporidium parvum, found in 7.7% of 4,922 samples, was the second most common parasite, exceeded only by Giardia lamblia which was found in 14.8% of the samples. The highest prevalence of cryptosporidium was found in children aged 6-11 months, whereas the prevalence of other enteric parasites increased with age. Cryptosporidiosis showed a marked seasonal variation, with peak prevalences found consistently at the beginning of or just before the rainy seasons, May through July. By contrast, no seasonality was found for the enteric parasites Giardia lamblia or Entamoeba histolytica. We conclude that Cryptosporidium parvum is an important pathogen in children with diarrhoea. PMID- 11732150 TI - A randomized controlled trial of the effect of antihelminthic treatment and micronutrient fortification on health status and school performance of rural primary school children. AB - Single interventions for helminthic infections and micronutrient deficiencies are effective, but it is not clear whether combined interventions will provide equal, additive or synergistic effects to improve children's health. The study objective was to determine the impact of single and combined interventions on nutritional status and scholastic and cognitive performance of school children. A double blind, randomized, placebo-controlled trial in 11 rural South African primary school randomly allocated 579 children aged between 8 and 10 years into six study groups, half of whom received antihelminthic treatment at baseline. The de-wormed and non-de-wormed arms were further divided into three groups and given biscuits, either unfortified or fortified with vitamin A and iron or with vitamin A only, given daily for 16 weeks. The outcome measures were anthropometric, micronutrient and parasite status, and scholastic and cognitive test scores. There was a significant treatment effect of vitamin A on serum retinol (p < 0.01), and the suggestion of an additive effect between vitamin A fortification and de-worming. Fortified biscuits improved micronutrient status in rural primary school children; vitamin A with de-worming had a greater impact on micronutrient status than vitamin A fortification alone and antihelminthic treatment significantly reduced the overall prevalence of parasite infection. The burden of micronutrient deficiency (anaemia, iron and vitamin A) and stunting in this study population was low and, coupled with the restricted duration of the intervention (16 weeks), might have limited the impact of the interventions. PMID- 11732151 TI - Use of radioiodine in adolescent Graves' disease. AB - Childhood hyperthyroidism is most commonly due to Graves' disease and often proves a therapeutic challenge. The high risk of relapse following medical management frequently necessitates definitive treatment. Radioactive iodine has been used for 60 years to treat hyperthyroidism. It is efficacious, cheap and safe. However, there is still some reluctance to use it in children. We report on the use and excellent short-term outcome of radioiodine therapy in five children with Graves' disease. PMID- 11732152 TI - Hirschsprung's disease in the newborn: experience in Zaria, Nigeria. AB - In a 10-year retrospective review of 15 newborns aged < or = 42 days presenting with Hirschsprung's disease, there were 12 boys and three girls aged 4-42 days (median 18 days). Twelve babies presented with complete intestinal obstruction. In 12 babies, there was a history of delayed passage of meconium (after 2-6 days). One baby each developed caecal and sigmoid perforation. Barium enemas in three babies without complete intestinal obstruction suggested Hirschsprung's disease in two. Following resuscitation, the two infants who had perforated had caecostomy and sigmoid repair with right transverse colostomy, respectively. One infant had ileostomy for total colonic aganglionosis associated with ileal atresia. All the others had initial diversion colostomy. Rectal biopsies confirmed Hirschsprung's disease in all the babies. The ileum was injured during colostomy in one case, requiring repair. Postoperative anastomotic leakage occurred in the infant with ileal injury and colostomy necrosis occurred in another infant. Five babies (33%) died, three from overwhelming infection (caecal perforation, sigmoid perforation, ileal injury), one from hypokalaemia (ileostomy) and one from an unidentified cause. Few cases of Hirschsprung's disease present in the newborn period in our environment and, when they do, they usually present with complete intestinal obstruction with high morbidity and mortality. PMID- 11732153 TI - Post-transfusion hepatitis C seroprevalence in Tanzanian children. AB - In Tanzania, children with malaria-associated anaemia are frequently given blood transfusions, and donor blood is not screened for hepatitis C virus (HCV) infection. To determine the seroprevalence of HCV infection in Tanzanian children previously transfused with blood, 184 children (92 transfused, 92 not transfused) aged between 15 and 59 months matched for age and sex were screened for HCV antibodies by the particle agglutination test using Serodia anti-HCV (Fujirebio Inc., Japan). The overall prevalence of HCV infection was 7.1% (13/184). HCV seropositivity was 5.4% (5/92) among children with a history of blood transfusion and 8.6% (8/92) among the non-transfused. There was no significant difference in the prevalence of HCV infection between transfused and non-transfused children. None of the factors investigated, such as gender, the nutrition and HIV serostatus of the children and the marital and education status of their mothers, was associated with HCV seropositivity. Further studies are recommended to identify the factors associated with HCV infection in Tanzanian children. PMID- 11732154 TI - Neonatal brucellosis and blood transfusion: case report and review of the literature. AB - Neonatal brucellosis following blood transfusion has not previously been reported. A premature male infant born at 24 weeks gestation developed low grade fever and decreased activity and showed poor weight gain at 45 weeks post menstrual age. Blood culture grew Brucella melitensis and the brucella antibody titre was positive. He received a 6-week course of septrin and rifampicin and made a full recovery. The infant had received a blood transfusion 5 days prior to his illness. The blood donor had symptoms suggestive of brucellosis, and it was suspected that the blood transfusion was the source of infection but this could not be confirmed as the donor was not traceable. It is suggested that, in areas endemic for brucellosis, prospective blood donors should be questioned about symptoms of brucellosis, and serological tests to screen for brucellosis might be indicated. PMID- 11732155 TI - Fast trains, slow boats, and the ancestry of the Polynesian islanders. AB - The question of the origins of the Polynesians has, for over 200 years, been the subject of adventure science. Since Captain Cook's first speculations on these isolated Pacific islanders, their language affiliations have been seen as an essential clue to the solution. The geographic and numeric centre of gravity of the Austronesian language family is in island Southeast Asia, which was therefore originally seen as their dispersal homeland. However, another view has held sway for 15 years, the 'out of Taiwan' model, popularly known as the 'express train to Polynesia'. This model, based on the combined evidence of archaeology and linguistics, proposes a common origin for all Austronesian-speaking populations, in an expansion of rice agriculturalists from south China/Taiwan beginning around 6,000 years ago. However, it is becoming clear that there is, in fact, little supporting evidence in favour of this view. Alternative models suggest that the ancestors of the Polynesians achieved their maritime skills and horticultural Neolithic somewhere between island Southeast Asia and Melanesia, at an earlier date. Recent advances in human genetics now allow for an independent test of these models, lending support to the latter view rather than the former. Although local gene flow occurring between the bio-geographic regions may have been the means for the dramatic cultural spread out to the Pacific, the immediate genetic substrate for the Polynesian expansion came not from Taiwan, but from east of the Wallace line, probably in Wallacea itself. PMID- 11732156 TI - Investigating urban geochemistry using Geographical Information Systems. AB - Geographical Information System (GIS) is an interactive digital extension of the two-dimensional paper map. Customised maps are created by the selection and aggregation of data from independent sources to assist studies in urban geochemistry. The metropolitan area of Wolverhampton, in the West Midlands, UK is used to illustrate the types of output that can be generated. These include: geographic and geological feature; geochemical data and land use. Multi-layered maps can be used to investigate spatial relationships, for example, between elevated concentrations of metals in soils and industrial land use. Such maps can also be used to assist the assessment of potential exposure of groundwater, ecosystems and humans using maps incorporating guideline values for metals in soils. PMID- 11732157 TI - Extracellular sensing and signalling pheromones switch-on thermotolerance and other stress responses in Escherichia coli. AB - The findings reviewed here overturn a major tenet of bacterial physiology, namely that stimuli which switch-on inducible responses are always detected by intracellular sensors, with all other components and stages in induction also being intracellular. Such an induction mechanism even applies to quorum-sensed responses, and some others which involve functioning of extracellular components, and had previously been believed to occur in all cases. In contrast, for the stress responses reviewed here, triggering is by a quite distinct process, pairs of extracellular components being involved, with the stress sensing component (the extracellular sensing component, ESC) and the signalling component, which derives from it and induces the stress (the extracellular induction component, EIC), being extracellular and the stimulus detection occurring in the growth medium. The ESCs and EICs can also be referred to as extracellular sensing and signalling pheromones, since they are not only needed for induction in the stressed culture, but can act as pheromones in the same region activating other organisms which fail to produce the extracellular component (EC) pair. They can also diffuse to other regions and there act as pheromones influencing unstressed organisms or those which fail to produce such ECs. The cross-talk occurring due to such interactions, can then switch-on stress responses in such unstressed organisms and in those which cannot form the ESC/EIC pair. Accordingly, the ESC/EIC pairs can bring about a form of intercellular communication between organisms. If the unstressed organisms, which are induced to stress tolerance by such extracellular components, are facing impending stress challenge, then the pheromonal activities of the ECs provide an early warning system against stress. The specific ESC/EIC pairs switch-on numerous responses; often these pairs are proteins, but non-protein ECs also occur and for a few systems, full induction needs two ESC/EIC pairs. Most of the above ECs needed for response induction are highly resistant to irreversible inactivation by lethal agents and conditions and, accordingly, many killed cultures still contain ESCs or EICs. If these killed cultures come into contact with unstressed living organisms, the ECs again act pheromonally, altering the tolerance to stress of the living organisms. It has been claimed that bacteria sense increased temperature using ribosomes or the DnaK gene product. The work reviewed here shows that, for thermal triggering of thermotolerance and acid tolerance in E. coli, it is ESCs which act as thermometers. PMID- 11732158 TI - Bacterial biofilms and human disease. AB - The term biofilm is used to denote a polymer-encased community of microbes which accumulates at a surface. Biofilms are responsible for a number of diseases of man and, because of the intrinsic resistance of these structures to antibiotics and host defence systems, such diseases are very difficult to treat effectively. The application of new microscopic and molecular techniques to biofilms has revolutionised our understanding of their structure, composition, organisation and activities. This review will describe the role that biofilms play in human disease and will outline our new millennial view of these complex and fascinating bacterial communities. PMID- 11732159 TI - Anchorage-dependent chick embryo fibroblasts synthesise DNA and proliferate when cultured in multilayered sheets suspended among glass fibres. AB - Chick embryo fibroblasts (CEFs) spontaneously form multicellular and multilayered sheets suspended on the network of glass fibres which are stabilized by fibronectin containing protein deposits located at cell-to-cell contacts. The cells situated within the sheets are surrounded by the neighbouring cells and their mechanical equilibrium is stabilised by intercellular "parabaric" effects. It was found that CEFs in the sheets retain relatively high mitotic activity corresponding to that observed in sparse monolayer cultures. These cells grew up to much higher local density than in confluent and contact-inhibited monolayer cultures and developed an abundance of microfilament bundles that terminated at vinculin-containing protein complexes. The results presented demonstrate that direct contact with solid substratum, cell-to-cell contacts, local cell density, and intercellular exchange of humoral factors are not directly involved in the density-dependent inhibition of growth observed in monolayer cultures. They also support the concepts concerning the role of mechanical equilibrium of cell membrane and sub-membranous cytoskeleton in the regulation of proliferation of non-transformed cells. PMID- 11732160 TI - Habitat of Paramecium biaurelia in Italy, the Island of Elba. AB - The new habitat of Paramecium biaurelia was found on the Island Elba. PMID- 11732161 TI - Species of the Paramecium aurelia complex in Japan. AB - In Japan, the presence of Paramecium sexaurelia was revealed for the first time, and a new habitat of P. tetraurelia was found there. PMID- 11732162 TI - Worker genetic diversity and infection by Nosema apis in honey bee colonies. AB - The hypothesis that parasites and pathogens select for polyandry in eusocial Hymenoptera was tested, using the honey bee Apis mellifera and its microsporidian parasite Nosema apis. Five honey bee colonies with low and five with high worker genetic diversity were infected with N. apis spores. At 54-56 days after inoculation, parasite spores in the workers' midguts were counted to determine whether there was a greater variation of infection intensity (spore counts per worker) in high-diversity colonies than in low-diversity ones. In all colonies there were two discrete sets of workers, with few or many parasite spores. To compare the variations of infection intensity between two colony groups, coefficients of variation were calculated for all workers examined, and for the slightly infected and strongly infected workers. The percentages of slightly infected workers in the low- and high-diversity groups were also compared. None of the comparisons between low- and high-diversity colonies showed significant differences, therefore no relation was found between honey bee workers' genetic diversity and their infection with N. apis. PMID- 11732163 TI - Anomalies of autogamy in Paramecium tetraurelia temperature sensitive mutant induced by heat treatment. AB - The run of autogamy in temperature sensitive mutant ts401, carrying also the mutation of trichocysts nd3a, at permissive (26 degrees C) and non-permissive temperature (35 degrees C) was studied and compared with this process in wild d4 2 stock of Paramecium tetraurelia. The effect of heat treatment on programmed nuclear and cortical events was investigated using cytological silver impregnation Fernandez-Galiano method and immunofluorescence technique with the application of two anti-alpha-tubulin antibodies. The appearance of cells with some large macronuclear fragments and an excess number of micronuclei suggests that nuclear differentiation is inhibited at restrictive temperature and that macronuclear regeneration takes place in this thermosensitive mutant. In such cells impaired oral reorganization was observed. After slight refeeding at the late autogamy, cells stopped at first postautogamous division were induced by heat treatment. The most striking feature of these abnormal divisions was the lack of a basal body duplication wave, which suggests that production of gamma tubulin, structural protein being the essential component of the microtubule organizing centres, is disturbed at high temperature. PMID- 11732164 TI - Chromosome numbers, sex determining systems, and patterns of the C heterochromatin distribution in 13 species of lace bugs (Heteroptera, Tingidae). AB - The karyotypes, sex chromosome systems, and male meiotic patterns in 13 species belonging to 10 genera of the family Tingidae were studied. Data on eleven species, one subgenus, and 5 genera are presented for the first time, and the chromosome formula of Acalypta parvula is revised. Karyotypes of all species included six pairs of autosomes. Most of the species displayed an XY sex chromosome system, in four species, belonging to genera of Acalypta and Kalama, the X0 system was found. Male meiosis is chiasmatic for autosomes. Sex chromosomes are achiasmatic and undergo pre-reductional meiosis. Using C-banding technique, for the first time constitutive heterochromatin was localized on chromosomes in all the species studied. The heterochromatin was found either in telomeres or in some species in interstitial locations, evidencing that a quite substantial redistribution of chromosome material within chromosomes might occur without fragmentations or fusions. In two species, a supernumerary (B) chromosome was found. In addition, the male reproductive system of four species was examined and the number of testicular follicles was determined as two per testis. PMID- 11732165 TI - Ag-banded karyotypes in six species of jewel-beetles (Coleoptera, Buprestidae). AB - The male karyotypes of six species of the buprestid genera Sphenoptera (two species) and Acmaeoderella (four species) were studied after Ag-banding. The following karyotypes were found in individual species: n = 19-23, with predominance of 23, in Sphenoptera scovitzi Fald., 2n = 24 (22 + Xyp) in Sphenoptera mesopotamica Mars., 2n = 18 (16 + Xyr) in Acmaeoderella flavofasciata Pill. & Mitt., Acmaeoderella gibbulosa Men., Acmaeoderella boryi Brulle, Acmaeoderella vetusta Men. In Sph. scovitzi, two chromosome pairs were found to display stable argentophilous blocks. In Sph. mesopotamica, NOR-bearing is one of the large autosomal bivalents. In all species of the genus Acmaeoderella, a nucleolus-like argentophilous body is placed between the X and y chromosomes. PMID- 11732166 TI - Cytogenetic investigations on seven Palaearctic weevil species (Coleoptera, Curculionidae). AB - Cytogenetic investigations were carried out on seven species of bisexual weevils from five subfamilies. The following numbers of chromosomes were found in individual species: 2n = 22, n male = 10 + Xyp in Ptochus porcellus Boh. (Otiorhynchinae); 2n = 40, n male = 19 + Xyp in Larinodontes turbinatus Gyll. (Cleoninae); 2n = 26, n male = 12 + Xyp in Curculio pellitus (Boh.) (Curculioninae); 2n = 30, n male = 14 + Xyp in Acalles camelus (F.), Acalles echinatus (Germ.), Acalles hypocrita Boh. (Cryptorhynchinae), and 2n = 40, n male = 19 + Xyp in Cionus hortulanus (Foucr.) (Mecininae). The heterochromosomes of all the examined species form, in the first meiotic metaphase, a parachute bivalent. The chromosome number and sex determining system of six species and two genera are described for the first time. PMID- 11732167 TI - Standard and C-banded meiotic karyotypes of Psylloidea (Sternorrhyncha, Homoptera, Insecta). AB - Meiotic karyotypes were studied in males of Craspedolepta sonchi (Foerster, 1848), Diaphorina chobauti Puton, 1898, D. lamproptera Burckhardt, 1981, Psylla hartigii Flor, 1861, Cacopsylla palmeni (Loew, 1878), C. hippophaes (Foerster, 1848), C. melanoneura (Foerster, 1868), C. pyricola (Foerster 1848), C. moscovita (Andrianova, 1848), Bactericera salicivora (Reuter, 1876), Trioza abdominalis Flor, 1861, T. lauri = Lauritrioza alacris (Flor, 1861). Karyotypes were 2n = 25 (24 + XO) in all species except B. salicivora with 2n = 26 (24 + neo-XY). Testes consisted of two follicles each in all species but P. hartigii with four follicular testes in males. The discussion covers the problems of chromosome numbers, sex-determining chromosome systems, B-chromosomes, patterns of C banding, testis structure, and spermatid development in Psylloidea. PMID- 11732168 TI - B chromosomes, translocation between B and autosomes, and C-heterochromatin polymorphism of the grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae) in Hokkaido, northern Japan. AB - Seven categories of B chromosomes found in the brachypterus grasshopper Podisma sapporensis from Hokkaido populations differ in structure, size, and C-band content. The interchange between B and one autosome from M3 and sporadically M7 was observed in most of the populations examined. Such an interaction between standard and non-standard chromosomal set provides an insight into the integration of supernumerary chromosome. In addition, C-heterochromatin polymorphism was also identified in male karyotypes in some populations. These facts indicate P. sapporensis is a highly polymorphic species from the cytogenetic point of view. PMID- 11732169 TI - The effect of dorsal raphe nucleus (DRN) lesions on the locomotor activity rhythm in mice. AB - The study employed electrical lesions of dorsal raphe nucleus (DRN) to determine the functional significance of those nuclei in the regulation of wheel-running activity rhythm in mice in light/dark (LD 12:12), constant light (LL), and constant dark (DD) conditions. The wheel-running records showed that raphe nucleus lesions resulted in few days' decrease in common activity and amplitude in LD. The activity phase was not compact but in fragmentary form, especially in DD condition. In some animals an earlier onset of activity after DRN lesion in LD was observed. In LL extension of the rhythm period occurred. Destruction of DRN only slightly modulates the wheel-running circadian rhythm in mice. PMID- 11732170 TI - Do germ line cells in Allacma fusca (Insecta, Collembola, Symphypleona) have a higher metabolic rate than somatic cells. AB - Stereological analysis of the ultrastructure of primordial germ cells (PGCs) and the somatic (ectoderm) cells in two developmental stages of embryos and freshly hatched juveniles of Allacma fusca have shown great differences in mitochondria volume density (vd) between the two types of cells. In younger embryos (migration phase of the PGCs) the vd of mitochondria in the cytoplasm of the PGCs is 74.64% higher than in the ectoderm cells. In older embryos, (PGCs in the gonads) the vd of mitochondria is 123% higher than the corresponding value for the somatic cells cytoplasm. In the juvenile the vd of mitochondria in the ectoderm cells grows twice but is still only 2/3 of the value for the PGCs. On the basis of papers describing a direct relationship between stereological and physiological results the authors conclude that the metabolism of the primordial germ cells during embryonic development of Allacma fusca is much higher than that of the somatic ones. If the above conclusion is correct, the results presented here confirm the "disposable soma theory" (Kirkwood & Holliday 1979). PMID- 11732171 TI - Structure of ovaries and oogenesis in Corydalidae and Chauliodidae (Insecta, Megaloptera). I. Architecture of adult ovarioles and previtellogenesis. AB - The results of histological and EM studies on the ovaries of three representatives of Megaloptera: Chauliodes pectinicornis, Nigronia fasciata (Chauliodidae), and Corydalus peruvianus Corydalidae) are presented. It is shown that the ovaries of all 3 investigated species are panoistic (secondary panoistic, = neopanoistic) and consist of numerous (more than a hundred) ovarioles that are differentiated into 3 well-defined regions: the terminal filament, the germarium, and the vitellarium. The germaria of adult females are apparently non-functional and contain germ and somatic cells in various stages of degeneration. The vitellaria are composed of 12-15 developing ovarian follicles (= oocytes surrounded by follicular cells) in a linear arrangement. In adult females these follicles can be classified into early previtellogenic, late previtellogenic, vitellogenic, and choriogenic. During early previtellogenesis oocyte nuclei (= germinal vesicles) contain single nucleolar masses. Histochemical analyses indicate that within the masses DNA as well as AgNOR proteins are present. During subsequent stages of the previtellogenic growth nucleolar masses gradually break down into smaller aggregations of coarse granular material, i.e. multiple nucleoli. In chauliodids the nucleoli are distributed evenly throughout the nucleoplasm while in the corydalid, C. peruvianus, they form a characteristic ring. The presented results are discussed in a phylogenetic context. PMID- 11732172 TI - Zooplankton in the artificial pond of the Botanical Garden of the Jagiellonian University in Krakow. AB - Paramecium caudatum and the predominant zooplanton organisms were registered in the studied pond of the Botanical Garden of the Jagiellonian University in Krakow. The finding of Stentor coeruleus indicates an improvement in the environmental conditions in Krakow, connected with the partial liquidation of the huge steelworks. PMID- 11732173 TI - A model of microbial contamination of a water reservoir. AB - A three year record of daily fecal coliform counts in a Massachusetts water reservoir has the appearance of an irregular time series punctuated by outbursts of varying duration. The pattern is described in terms of a probabilistic model where the fluctuations in the 'regular' and 'explosive' regimes are governed by two sets of probabilities. It has been assumed that the random oscillations has a lognormal distribution, and that once an explosion threshold has been exceeded the increments or decrements in the population size have fixed probability distributions. The threshold for triggering an outburst was estimated by examining the randomness of the autocorrelation function of the record after it is filtered to eliminate peaks of progressively increasing magnitude. Once the threshold has been identified, the mean and standard deviation of the underlying lognormal distribution could be estimated directly from remains found in the record after all the peaks were removed. The probabilities of an increment and decrement during the outbursts and their relative magnitudes could also be estimated using simple formulas. These estimated parameter values were then used to generate realistic records with known threshold levels, which were subsequently used to assess the procedure's feasibility and sensitivity. PMID- 11732174 TI - One-vesicle hypothesis for neurotransmitter release: a possible molecular mechanism. AB - Neurotransmitter-containing vesicles are clustered in release sites. Although a given site can contain tens of vesicles, there is evidence that under a wide range of conditions, following an action potential, rarely is more than one vesicle released from each site. Such findings led to the one vesicle hypothesis, for which this paper suggests a molecular mechanism. The release of a vesicle from a site provides a transient high concentration of transmitter in that site. It is proposed here that the local high transmitter concentration interrupts further vesicle releases from the same release site. The suggested mechanism for this 'release interruption' is based on a theory of release control by the authors wherein inhibitory transmitter autoreceptors play a central role. (That transmitter binding to these autoreceptors can inhibit release on a fast time scale has recently been shown experimentally.) A detailed kinetic scheme is presented for the proposed mechanism. Stochastic simulations of this scheme demonstrate how the mechanism accounts for the one vesicle hypothesis. In agreement with recent experiments, the simulations also show that changes in conditions that affect the release process can cause frequent release of more than one vesicle per site. PMID- 11732175 TI - Modelling the natural history of HIV infection in individuals and its epidemiological implications. AB - The variation of viraemia in the natural course of HIV infection is expected to have major influence on the probability of transmission and, consequently, on the epidemiology of HIV/AIDS. In this paper we propose a model which takes into account the time evolution of HIV viraemia (measured as HIV-RNA copies per ml of blood) in an infected individual and its impact on the threshold for the establishment of an endemic level, and mainly on the relative contribution of each of the clinical phases of the infection to the total transmission of HIV per infected individual. We consider that an infected individual passes through three phases of viraemia. The first phase, which lasts for 6-7 weeks, is characterized by very high viraemia. In the second phase, which lasts about 10 years, the viraemia is much lower, increasing again in the last phase, which lasts up to two years, and ends in full-blown AIDS. We show that the relative contribution of each phase to the total transmission of HIV is very sensitive to the model we assume for the dependence of the transmissibility of HIV on the viral load. For instance, if we assume that transmissibility is proportional to the decimal logarithm of viraemia, then the second phase predominates always. Due to the epidemiological importance of this fact, it is clear that further improvement on virological research to better understand the dependence of HIV transmissibility on the viral concentration in biological fluids is necessary. PMID- 11732176 TI - Estimating the size of the olfactory repertoire. AB - The concept of shape space, which has been successfully implemented in immunology, is used here to construct a model for the discrimination power of the olfactory system. Using reasonable assumptions on the behaviour of the biological system, we are able to estimate the number of distinct olfactory receptor types. Our estimated value of around 1000 receptor types is in good agreement with experimental data. PMID- 11732177 TI - Model of droplet dynamics in the Argentine ant Linepithema Humile (Mayr). AB - The formation of droplets of ants Linepithema humile (Mayr) is observed under certain experimental conditions: a fluctuating aggregate forms at the end of a rod and a droplet containing up to 40 ants eventually falls down. When the flux of incoming ants is sufficient, this process can continue for several hours, leading to the formation and fall of tens of droplets. Previous work indicates that the time series of drop-to-drop intervals may result from a nonlinear low dimensional dynamics, and the interdrop increments exhibit long-range anticorrelations. A model of aggregation and droplet formation, based on experimental observations, is introduced and shown to reproduce these properties. PMID- 11732178 TI - Critical conditions for phytoplankton blooms. AB - We motivate and analyse a reaction-advection-diffusion model for the dynamics of a phytoplankton species. The reproductive rate of the phytoplankton is determined by the local light intensity. The light intensity decreases with depth due to absorption by water and phytoplankton. Phytoplankton is transported by turbulent diffusion in a water column of given depth. Furthermore, it might be sinking or buoyant depending on its specific density. Dimensional analysis allows the reduction of the full problem to a problem with four dimensionless parameters that is fully explored. We prove that the critical parameter regime for which a stationary phytoplankton bloom ceases to exist, can be analysed by a reduced linearized equation with particular boundary conditions. This problem is mapped exactly to a Bessel function problem, which is evaluated both numerically and by asymptotic expansions. A final transformation from dimensionless parameters back to laboratory parameters results in a complete set of predictions for the conditions that allow phytoplankton bloom development. Our results show that the conditions for phytoplankton bloom development can be captured by a critical depth, a compensation depth, and zero, one or two critical values of the vertical turbulent diffusion coefficient. These experimentally testable predictions take the form of similarity laws: every plankton-water-light-system characterized by the same dimensionless parameters will show the same dynamics. PMID- 11732179 TI - Two-category model of task allocation with application to ant societies. AB - In many network models of interacting units such as cells or insects, the coupling coefficients between units are independent of the state of the units. Here we analyze the temporal behavior of units that can switch between two 'category' states according to rules that involve category-dependent coupling coefficients. The behaviors of the category populations resulting from the asynchronous random updating of units are first classified according to the signs of the coupling coefficients using numerical simulations. They range from isolated fixed points to lines of fixed points and stochastic attractors. These behaviors are then explained analytically using iterated function systems and birth-death jump processes. The main inspiration for our work comes from studies of non-hierarchical task allocation in, e.g., harvester ant colonies where temporal fluctuations in the numbers of ants engaged in various tasks occur as circumstances require and depend on interactions between ants. We identify interaction types that produce quick recovery from perturbations to an asymptotic behavior whose characteristics are function of the coupling coefficients between ants as well as between ants and their environment. We also compute analytically the probability density of the population numbers, and show that perturbations in our model decay twice as fast as in a model with random switching dynamics. A subset of the interaction types between ants yields intrinsic stochastic asymptotic behaviors which could account for some of the experimentally observed fluctuations. Such noisy trajectories are shown to be random walks with state dependent biases in the 'category population' phase space. With an external stimulus, the parameters of the category-switching rules become time-dependent. Depending on the growth rate of the stimulus in comparison to its population dependent decay rate, the dynamics may qualitatively differ from the case without stimulus. Our simple two-category model provides a framework for understanding the rich variety of behaviors in network dynamics with state-dependent coupling coefficients, and especially in task allocation processes with many tasks. PMID- 11732180 TI - An algebraic-combinatorial model for the identification and mapping of biochemical pathways. AB - We develop the mathematical machinery for the construction of an algebraic combinatorial model using Petri nets to construct an oriented matroid representation of biochemical pathways. For demonstration purposes, we use a model metabolic pathway example from the literature to derive a general biochemical reaction network model. The biomolecular networks define a connectivity matrix that identifies a linear representation of a Petri net. The sub-circuits that span a reaction network are subject to flux conservation laws. The conservation laws correspond to algebraic-combinatorial dual invariants, that are called S- (state) and T- (transition) invariants. Each invariant has an associated minimum support. We show that every minimum support of a Petri net invariant defines a unique signed sub-circuit representation. We prove that the family of signed sub-circuits has an implicit order that defines an oriented matroid. The oriented matroid is then used to identify the feasible sub-circuit pathways that span the biochemical network as the positive cycles in a hyper digraph. PMID- 11732182 TI - Behavior of plastid nucleoids during male gametogenesis in Plumbago auriculata. AB - We characterized the behavior of plastid (pt) and mitochondrial (mt) nucleoids during male gametogenesis in Plumbago auriculata in three dimensions. The behavior of pt-nucleoids and mt-nucleoids differed throughout male gametogenesis. Pt-nucleoids were distributed in a characteristic manner in three stages: in the early microspore, pt-nucleoids assemble around cell nucleus; in the mid generative cell, pt-nucleoids gather at the internal side of the pollen; in the late-generative cell, pt-nucleoids aggregation turns its pole to the external side of the pollen. We also studied organelle nucleoids in the egg and the central cell by a method in which semi-thick sections of resin-embedded anthers and ovaries were observed by confocal laser scanning microscopy. The number of pt nucleoids in the sperm cell did not differ significantly from that in the egg. These results suggest that the behavior of DNA-containing organelles is regulated strictly during male gametogenesis in P. auriculata, and that a biparental inheritance of plastids in the Plumbago embryo is more favored than was previously thought. PMID- 11732183 TI - Immunogold localization of a transglutaminase related to grana development in different maize cell types. AB - A comparative study of the subcellular localization of a plant transglutaminase (TGase; EC 2.3.2.13) in various in vivo and in vitro maize cell types was carried out with a polyclonal antibody raised against a 58 kDa TGase purified from Helianthus tuberosus leaves. Immunocytochemical staining, followed by electron microscopy, showed that this enzyme was markedly present in the grana-appressed thylakoids of mature chloroplasts of the light-exposed cells. Moreover, during embryogenic callus chloroplast differentiation, the abundance of TGase in the grana-appressed thylakoids depended on the degree of grana development and was greater than in mature leaf chloroplasts. In addition to the 58 kDa form, two other forms of the protein (of 77 and 34 kDa) were obtained by Western blot. The 77 kDa form might correspond to the inactive form and was immunodetected in dense vesicles observed in dark-grown embryogenic callus cells. In adult leaves, the enzyme was also markedly present in the grana-appressed thylakoids of the mesophyll cell chloroplasts, though very scarce and dispersed in the bundle sheath cell chloroplasts (which do not contain grana). The concordance of these localizations with those described for the light-harvesting antenna proteins of the photosystem II suggests that it is possible that this TGase has a functional role in photosynthesis, perhaps modulating the photosynthetic efficiency and the absorption of excess light by means of polyamine conjugation to the antenna proteins. PMID- 11732181 TI - Cyclin/Cdk complexes: their involvement in cell cycle progression and mitotic division. AB - DNA replication and mitosis are dependent on the activity of cyclin-dependent protein kinase (CDK) enzymes, which are heterodimers of a catalytic subunit with a cyclin subunit. Cyclin binding to specific individual proteins is thought to provide potential substrates to Cdk. Protein binding by cyclins is assessed in terms of its mechanisms and biological significance, using evidence from diverse organisms including substrate specificity in animal Cdk enzymes containing D-, A , and B-type cyclins and extensive cyclin gene manipulations in yeasts. Assembly of protein complexes with cyclin/Cdk is noted and the capacity of the cyclin dependent kinase subunit Cks, in such complex, to extend the range of Cdk substrates is documented and discussed in terms of cell cycle regulation. Cell cycle progression involves changing abundance of individual cyclins, due to changing rates of their transcription or proteolysis, with consequent changes in the substrates of CDK through the cell cycle. Some overlap of the functions of individual cyclins in vivo has been identified by cyclin deletions and is suggested to follow a pattern in which cyclins can commonly complete functions initiated by the preceding cyclins well enough to preserve viability as groups of cyclins are removed by proteolysis. Cyclin accumulation is particularly important in terminating the G1 phase, when it raises CDK activity and starts events leading to DNA replication. It is suggested that plants share this mechanism. The distribution of cyclins and Cdk in maize root tip cells during mitosis and cytokinesis indicates the presence of Cdk1 (Cdc2a) and cyclin CycB1zm;2 at the mature and disassembling preprophase band and the presence of CycB1zm;2 at condensing and condensed chromosomes. Both observations correlate with the earlier-reported capacity of injected metaphase cyclin/CDK to accelerate preprophase band disassembly and chromosome condensation and with observations of the location of Cdk and cyclins in other laboratories. Additionally CycB1zm;2 is seen at the nuclear envelope during its breakdown, which correlates with an acceleration of the process by injected metaphase cyclin B/CDK. A phenomenon possibly unique to the plant kingdom is the persistence of mitotic cyclins after anaphase. Participation of cyclins in cytokinesis is indicated by the concentration of the mitotic cyclin CycA1;zm;1 at the phragmoplast. It is suggested that cyclins have a general function of spatially focusing Cdk activity and that in the plant cell the concentrations of cyclins are important mediators of CDK activity at the cytoskeleton, chromosomes, spindle, nuclear envelope, and phragmoplast. PMID- 11732184 TI - Day-to-night variations of cytoplasmic pH in a crassulacean acid metabolism plant. AB - In crassulacean acid metabolism (CAM) large amounts of malic acid are redistributed between vacuole and cytoplasm in the course of night-to-day transitions. The corresponding changes of the cytoplasmic pH (pHcyt) were monitored in mesophyll protoplasts from the CAM plant Kalanchoe daigremontiana Hamet et Perrier by ratiometric fluorimetry with the fluorescent dye 2',7'-bis-(2 carboxyethyl)-5-(and-6-)carboxyfluorescein as a pHcyt indicator. At the beginning of the light phase, pHcyt was slightly alkaline (about 7.5). It dropped during midday by about 0.3 pH units before recovering again in the late-day-to-early dark phase. In the physiological context the variation in pHcyt may be a component of CAM regulation. Due to its pH sensitivity, phosphoenolpyruvate carboxylase appears as a likely target enzyme. From monitoring delta pHcyt in response to loading the cytoplasm with the weak acid salt K-acetate a cytoplasmic H(+)-buffer capacity in the order of 65 mM H+ per pH unit was estimated at a pHcyt of about 7.5. With this value, an acid load of the cytoplasm by about 10 mM malic acid can be estimated as the cause of the observed drop in pHcyt. A diurnal oscillation in pHcyt and a quantitatively similar cytoplasmic malic acid is predicted from an established mathematical model which allows simulation of the CAM dynamics. The similarity of model predictions and experimental data supports the view put forward in this model that a phase transition of the tonoplast is an essential functional element in CAM dynamics. PMID- 11732185 TI - Pollen hydration status at dispersal: cytophysiological features and strategies. AB - The aim of this paper is to draw attention to partially hydrated pollen, namely, pollen grains having a high water content (> 30%); this type of pollen is more frequent than previously thought. Various cyto-physiological strategies are used to retain water during exposure and dispersal such as cytoplasm carbohydrates; in the absence of such strategies, fast pollination must be ensured, because uncontrolled loss of water leads to pollen death. On the other hand, a state of partial hydration allows a fast tube emission (even within 3-5 min). Several methods for determining the hydration status of pollen at anthesis are proposed. PMID- 11732187 TI - Distribution of splicing proteins and putative coiled bodies during pollen development and androgenesis in Brassica napus L. AB - Small nuclear ribonucleoprotein particles (snRNPs) are subunits of splicing complexes, which show a transcription-dependent localization pattern. We have analyzed the labelling pattern of snRNPs during pollen development and microspore and pollen embryogenesis in Brassica napus with an antibody which recognizes protein D of U1, U2, U4, U5, and U6 snRNPs. It was found that nuclei were labelled almost uniformly for snRNPs in microspores and young bicellular pollen. In the generative nuclei of late-bicellular pollen and in the vegetative nuclei and sperm nuclei of mature pollen no snRNPs could be detected. The snRNP-positive nuclei contained mostly one or two brightly labelled nuclear bodies, most likely coiled bodies, often closely related to the nucleolus. These nuclear bodies increased in size from 0.5 micron in nuclei of young microspores up to 2 microns in nuclei of late microspores and the vegetative nucleus of early-bicellular pollen. Also their number increased during these developmental stages. After induction of embryogenesis the size of the coiled bodies decreased to about 0.5 micron and in several occasions the coiled body was found free in the nucleoplasm, away from the nucleolus. The results support the idea that the size and number of coiled bodies coincide with changes in general nuclear activity. They also indicate that, in nuclei of Brassica napus, at least assembly and disassembly of coiled bodies takes place in the nucleoplasm, whereas mature coiled bodies are located adjacent to the nucleolus. PMID- 11732186 TI - cDNA isolation, characterization, and protein intracellular localization of a katanin-like p60 subunit from Arabidopsis thaliana. AB - Katanin, a heterodimeric protein with ATP-dependent microtubule-severing activity, localizes to the centrosome in animal cells. Widespread occurrence is suspected as several species contain homologs to the katanin p60 subunit. Recently we isolated an Arabidopsis thaliana cDNA with significant identity to the p60 subunit of sea urchin katanin. Like p60, the encoded protein is a member of the AAA superfamily of ATPases, containing the Walker ATP binding consensus and the signature AAA minimal consensus sequences within a single larger AAA/CAD amino acid motif. Phylogenetic analysis placed the encoded protein in the AAA subfamily of cytoskeleton-interactive proteins, where it formed a strongly supported clade with 4 other members identified as katanin p60 subunits. The clone was named AtKSS (Arabidopsis thaliana katanin-like protein small subunit). Western blots, performed using a polyclonal antibody raised against recombinant AtKSS, revealed AtKSS is present in protein extracts of all Arabidopsis organs examined. To evaluate potential interactions between AtKSS and the cytoskeleton, the intracellular localization of AtKSS was correlated with that of tubulin. AtKSS was found in perinuclear regions during interphase, surrounding the spindle poles during mitosis, but was absent from the preprophase band and phragmoplast microtubule arrays. These data support the thesis that AtKSS is an Arabidopsis homolog of the p60 subunit of katanin. Its cell cycle-dependent distribution is consistent with microtubule-severing activity, but additional studies will better define its role. PMID- 11732188 TI - Spatiotemporal relationships between growth and microtubule orientation as revealed in living root cells of Arabidopsis thaliana transformed with green fluorescent-protein gene construct GFP-MBD. AB - Arabidopsis thaliana plants were transformed with GFP-MBD (J. Marc et al., Plant Cell 10: 1927-1939, 1998) under the control of a constitutive (35S) or copper inducible promoter. GFP-specific fluorescence distributions, levels, and persistence were determined and found to vary with age, tissue type, transgenic line, and individual plant. With the exception of an increased frequency of abnormal roots of 35S GFP-MBD plants grown on kanamycin-containing media, expression of GFP-MBD does not appear to affect plant phenotype. The number of leaves, branches, bolts, and siliques as well as overall height, leaf size, and seed set are similar between wild-type and transgenic plants as is the rate of root growth. Thus, we conclude that the transgenic plants can serve as a living model system in which the dynamic behavior of microtubules can be visualized. Confocal microscopy was used to simultaneously monitor growth and microtubule behavior within individual cells as they passed through the elongation zone of the Arabidopsis root. Generally, microtubules reoriented from transverse to oblique or longitudinal orientations as growth declined. Microtubule reorientation initiated at the ends of the cell did not necessarily occur simultaneously in adjacent neighboring cells and did not involve complete disintegration and repolymerization of microtubule arrays. Although growth rates correlated with microtubule reorientation, the two processes were not tightly coupled in terms of their temporal relationships, suggesting that other factor(s) may be involved in regulating both events. Additionally, microtubule orientation was more defined in cells whose growth was accelerating and less stringent in cells whose growth was decelerating, indicating that microtubule-orienting factor(s) may be sensitive to growth acceleration, rather than growth per se. PMID- 11732189 TI - Localization of cell wall polysaccharides in nonarticulated laticifers of Asclepias speciosa Torr. AB - Asclepias speciosa Torr. has latex-containing cells known as nonarticulated laticifers. In stem sections of this species, we have analyzed the cell walls of nonarticulated laticifers and surrounding cells with various stains, lectins, and monoclonal antibodies. These analyses revealed that laticifer walls are rich in (1-->4) beta-D-glucans and pectin polymers. Immunolocalization of pectic epitopes with the antihomogalacturonan antibodies JIM5 and JIM7 produced distinct labeling patterns. JIM7 labeled all cells including laticifers, while JIM5 only labeled mature epidermal cells and xylem elements. Two antibodies, LM5 and LM6, which recognize rhamnogalacturonan I epitopes distinctly labeled laticifer walls. LM6, which binds to a (1-->5) alpha-arabinan epitope, labeled laticifer walls more intensely than walls of other cells. LM5, which recognizes a (1-->4) beta-D galactan epitope, did not label laticifer segments at the shoot apex but labeled more mature portions of laticifers. Also the LM5 antibody did not label cells at the shoot apical meristem, but as cells grew and matured the LM5 epitope was expressed in all cells. LM2, a monoclonal antibody that binds to beta-D glucuronic acid residues in arabinogalactan proteins, did not label laticifers but specifically labeled sieve tubes. Sieve tubes were also specifically labeled by Ricinus communis agglutinin, a lectin that binds to terminal beta-D-galactosyl residues. Taken together, the analyses conducted showed that laticifer walls have distinctive cytochemical properties and that these properties change along the length of laticifers. In addition, this study revealed differences in the expression of pectin and arabinogalactan protein epitopes during shoot development or among different cell types. PMID- 11732190 TI - Architecture of the Golgi apparatus of a scale-forming alga: biogenesis and transport of scales. AB - Mechanisms of transport of secretory products across the Golgi apparatus (GA) as well as of scale formation in prymnesiophytes have remained controversial. We have used a quantitative morphological approach to study formation and transport of scales across the GA in haploid cells of Pleurochrysis sp. The GA of these cells differs from the GA of higher plants in at least six morphological characteristics. Our results show that scales form in the trans-Golgi network (TGN) and transit the TGN in heretofore unrecognized prosecretory vesicles. Prosecretory vesicles differentiate into secretory vesicles prior to exocytosis of scales to the cell surface. Because prosecretory vesicles are only fragments of TGN cisternae, the classical model of cisternal progression is not a valid mechanism of transport in this alga. TGN transport vesicles are also involved in scale formation; however, the role of tubular connections between cisternae of a single stack-TGN unit is not clear. The relationship of two morphological types of cisternal dilations to a membrane-associated, bottlebrush-shaped macromolecule of novel morphology suggests a new hypothesis for the biogenesis of scales. PMID- 11732191 TI - Primary and secondary plasmodesmata: structure, origin, and functioning. AB - In the multicellular organisms of higher plants, plasmodesmata provide pathways for intimate symplasmic communication between neighboring cells. The arguments summarized in the present review demonstrate that plasmodesmata are diverse and highly dynamic structures. Differences in the plasmodesmal origin and modifications of the plasmodesmal structure and functioning at the various cell interfaces are the basic means which give rise to a complicated and flexibile symplasmic network. This complex communication system is discussed to serve a significant role in the coordinated development and in the concerted physiological functioning of the cells within the plant tissues, organs, and organisms. PMID- 11732192 TI - The vacuole as central element of the lytic system and sink for lipid droplets in maturing appressoria of Magnaporthe grisea. AB - Histochemical and ultrastructural studies were carried out on a wild-type strain (Guy11) and a melanin-deficient mutant (buf1) of the rice-blast pathogen, Magnaporthe grisea (= Pyricularia oryzae), in order to investigate the destination of lipid storage reserves during appressorium development. Lipid droplets were abundant in conidia and were mobilised upon germination, accumulating in the appressorial hook which developed at the tip of each germ tube. Following the formation of a septum at the base of the nascent appressorium, one or a few closely appressed central vacuoles became established and were observed to enlarge in the course of appressorium maturation. On unyielding artificial surfaces such as glass or plastic, appressoria matured to completion within 36-48 h, by which time the enlarged vacuole filled most of the inside volume of the appressorium. Light and transmission electron microscopical observations revealed that the lipid droplets entered the vacuole by autophagocytosis and were degraded therein. Histochemical approaches confirmed the vacuole as the key lytic element in maturing appressoria. Endocytosis of a vital dye, Neutral Red, progressed via endosomes which migrated into the vacuole and lysed there, releasing their dye content into the vacuolar lumen. Furthermore, activity of the lysosomal marker enzyme, acid phosphomonoesterase, was strongly localised in the vacuole at all stages of appressorium maturation. It is therefore envisaged that vacuoles are involved in the degradation of lipid storage reserves which may act as sources of energy and/or osmotically active metabolites such as glycerol, which generate the very high turgor pressure known to be crucial for penetration of hard surfaces. On softer surfaces such as onion epidermis, appressoria of M. grisea were able to penetrate before degradation of lipid droplets had been completed. PMID- 11732193 TI - Microtubule dynamics are involved in stomatal movement of Vicia faba L. AB - To obtain a full picture of microtubule (MT) behavior during the opening and closure of guard cells we have microinjected living guard cells of Vicia faba with fluorescent tubulin, examined fine detail by freeze shattering fixed cells, and used drug treatments to confirm aspects of MT dynamics. Cortical MTs in fully opened guard cells are transversely oriented from the ventral wall to the dorsal wall. When the stomatal aperture was decreased by darkness, these MTs became twisted and patched and broken down into diffuse fragments when stomata were closed. When the closed stomata were opened in response to light, the MTs in guard cells changed from the diffused, transitional pattern back to one in which MTs are transversely oriented from stomatal pore to dorsal wall. This observation indicates a linkage between these MT changes and stomatal movement. To confirm this, we used the MT-stabilizing agent taxol and the MT-depolymerizing herbicide oryzalin and observed their effects on the stomatal aperture and MT dynamics. Both drugs suppressed light-induced stomatal opening and dark-induced closure. MTs are known to be necessary for maintaining the static kidney shape of guard cells; the present data now show that the dynamic properties of polymeric tubulin accompany changes in shape with stomatal movement and may be functionally involved in stomatal movement. PMID- 11732194 TI - Tunicamycin and Brefeldin A induce in plant cells a programmed cell death showing apoptotic features. AB - The recent identification of DAD (defender against apoptotic death) gene in plants suggests that the N-linked glycosylation of proteins could be an important control point of plant programmed cell death. In this paper we describe the effects of Tunicamycin, an inhibitor of N-linked protein glycosylation, and Brefeldin A, an inhibitor of protein trafficking from the Golgi apparatus, on sycamore (Acer pseudoplatanus L.) cell cultures. These two chemicals proved able to induce a strong acceleration of the cell death; changes in cell and nucleus morphology; an increase in DNA fragmentation, detectable by a specific immunological reaction; and the presence of oligonucleosomal-size fragments (laddering) in DNA gel electrophoresis. Moreover, Brefeldin A, but not Tunicamycin, strongly stimulated the production of hydrogen peroxide. These results indicate that also in plants chemicals interfering with the activities of endoplasmic reticulum and of Golgi apparatus strongly induce a form of programmed cell death showing apoptotic features. PMID- 11732195 TI - Electroporation modulates the embryogenic responses of asparagus (Asparagus officinalis L.) microspores. AB - Microspores of three genotypes of Asparagus officinalis L. were mechanically isolated without affecting their viability and were submitted to electric fields in order to modulate their competence for embryogenesis. When a constant pulse length and various field strengths (250, 500, 750, 1000, 1500, and 2000 V/cm) were tested, the viability of electro-treated microspores decreased as the field strength increased, for all genotypes. Conversely, the embryogenic competence was genotype dependent and was enhanced by low voltages for two clones when microspores were cultured in the presence of auxin. When the effect of pulse duration was studied, despite a strong genotype effect on responses, a short pulse coupled with a low voltage appeared to improve the competence for proembryo formation compared with nonelectroporated microspores, while longer pulses significantly improved microspore division. PMID- 11732197 TI - Poly(A)+ RNA during vegetative development of Acetabularia peniculus. AB - In the juvenile stage, the diploid giant-celled green algae Acetabularia spp. are differentiated into an upright stalk and an irregularly branched rhizoid. Early amputation and grafting experiments as well as biochemical and molecular analyses have shown that mRNA (as poly(A)+ RNA) is continuously supplied from the primary nucleus in the rhizoid and accumulates in the stalk apex. In the present study, localization of poly(A)+ RNA in the juvenile stage of the Acetabularia peniculus was investigated by fluorescent in situ hybridization using oligo(dT) as a probe. The signal was localized in the apical cytoplasm and, in addition, multiple longitudinal striations throughout the stalk and rhizoid cytoplasm. A large portion of the poly(A)+ RNA striations exhibited structural polarity, broadened at one end and gradually thinned toward the other end. Some of the striations in the rhizoid cytoplasm were continuous with a zone of signal in the area of the perinuclear rim. The poly(A)+ RNA striations were associated with thick bands of longitudinal actin bundles which run through the entire length of the stalk. Cytochalasin D caused fragmentation of the actin bundles and irregular distribution of the fluorescent signal. We suggest that the poly(A)+ RNA striations constitute a hitherto unknown form of packaged mRNA that is transported over large distances along the actin cytoskeleton to be stored and expressed in the growing apex. PMID- 11732196 TI - Effects of tissue-preparation-induced callose synthesis on estimates of plasmodesma size exclusion limits. AB - Plasmodesmata are often characterised by their size exclusion limit (SEL), which is the molecular weight of the largest dye, introduced by microinjection, that will move from cell to cell. In this study, we investigated whether commonly used techniques for isolation and manipulation of tissues, and microinjection of fluorescent dyes, affected the SEL, and whether any such effects could be ameliorated by inhibiting callose deposition. We examined young root epidermal cells of Arabidopsis thaliana and staminal hair cells of Tradescantia virginiana, two tissues often used in experiments on symplastic transport. Transport in root tips dissected from the main plant body and in stamen hairs removed from the base of the stamen filament was compared with transport in undissected roots and stamen hairs attached to the base of the filament, respectively. Tissues were microinjected with fluorescent dyes (457 Da to > 3 kDa) with or without prior incubation in the callose deposition inhibitors 2-deoxy-D-glucose or aniline blue fluorochrome. In both tissues, dissection reduced the SEL, which was largely prevented by prior incubation in 2-deoxy-D-glucose but not by incubation in aniline blue fluorochrome. Thus, standard methods for tissue preparation can cause sufficient callose deposition to reduce cell-to-cell transport, and this needs to be considered in studies employing microinjection. Introduction of the dyes by pressure injection rather than iontophoresis decreased the SEL in A. thaliana but increased it in T. virginiana, showing that these two injection techniques do not necessarily give identical results and that plasmodesmata in different tissues may respond differently to similar experimental procedures. PMID- 11732198 TI - Chaotic behavior in the locomotion of Amoeba proteus. AB - The locomotion of Amoeba proteus has been investigated by algorithms evaluating correlation dimension and Lyapunov spectrum developed in the field of nonlinear science. It is presumed by these parameters whether the random behavior of the system is stochastic or deterministic. For the analysis of the nonlinear parameters, n-dimensional time-delayed vectors have been reconstructed from a time series of periphery and area of A. proteus images captured with a charge coupled-device camera, which characterize its random motion. The correlation dimension analyzed has shown the random motion of A. proteus is subjected only to 3-4 macrovariables, though the system is a complex system composed of many degrees of freedom. Furthermore, the analysis of the Lyapunov spectrum has shown its largest exponent takes positive values. These results indicate the random behavior of A. proteus is chaotic and deterministic motion on an attractor with low dimension. It may be important for the elucidation of the cell locomotion to take account of nonlinear interactions among a small number of dynamics such as the sol-gel transformation, the cytoplasmic streaming, and the relating chemical reaction occurring in the cell. PMID- 11732199 TI - Cellulose in the house of the appendicularian Oikopleura rufescens. AB - By electron diffraction analysis, highly crystalline cellulose I beta was found in the house (a special structure in which the tunicate lives) of the appendicularian Oikopleura rufescens. Cellulose microfibrils 20 nm in width were observed in a random array or highly organized with rectangular spacing of 2 to 10 microns in the house. The bundled cellulose microfibrils formed in the inlet filters, which are highly ordered meshwork structures. This paper provides the first account of the existence of cellulose in the house of an appendicularian. Our findings showed that the house and tunic are homologous tissues among the tunicates, and that the common ancestor of the tunicates (ascidians, thaliaceans, and appendicularians) already possessed cellulose-biosynthetic ability. PMID- 11732200 TI - A dinoflagellate mutant with higher frequency of multiple fission. AB - The dinoflagellate Crypthecodinium cohnii Biecheler propagates by both binary and multiple fission. By a newly developed mutagenesis protocol based on using ethyl methanesulfonate and a cell size screening method, a cell cycle mutant, mf2, was isolated with giant cells which predominantly divide by multiple fission. The average cell size of the mutant mf2 is larger than the control C. cohnii. Cell cycle synchronization experiments suggest that mutant mf2, when compared with the control strain, has a prolonged G1 phase with a corresponding delay of the G2 + M phase. PMID- 11732201 TI - Ultrastructural effects of cellulose biosynthesis inhibitor herbicides on developing cotton fibers. AB - Cotton fibers are often utilized as a model system to investigate cellulose biosynthesis and cell wall elongation. In this study, we grew cotton fibers in vitro, with ovules dissected at day zero post anthesis as the explant source, in the presence of three herbicides that inhibit cellulose biosynthesis. Cultures were sampled for electron microscopy and immunocytochemistry 1-2 days after beginning the treatments. After dichlobenil treatment, the fibers were much shorter than the controls and assumed a variety of abnormal shapes, from shortened versions of the control fiber to nearly spherical. The inner layers of the fiber wall often contained juxtaposed electron-translucent and -transparent areas; this layer reacted strongly with antibodies to callose. Cellulase-gold labeling in these newly developed fibers grown in the presence of dichlobenil was present at only about 3% of the control labeling. After treatment with either isoxaben or flupoxam, the fibers assumed spherical shapes and frequently (more than 60% of fibers) exhibited a new cell plate within the fiber, indicating that cell division had occurred, a process that rarely occurred in the controls. Unlike the dichlobenil-treated fibers, fibers grown in the presence of isoxaben or flupoxam contained an extensive accumulation of chiefly deesterified pectins, replacing the entire wall with an elaborated version of the pectin sheath found in control cotton fibers. These data indicate that all three herbicides are effective disrupters of cellulose biosynthesis and cause radical changes in cell wall structure and composition. Moreover, these data indicate that the composition of the walls may influence indirectly cell cycle kinetics, keeping these fiber cells in a more meristematic mode. PMID- 11732202 TI - Subcellular organization of N2-fixing nodules of cowpea (Vigna unguiculata) supplied with silicon. AB - Provision of silicon (0, 0.048, 0.096, 0.24, 0.48, and 0.96 g/l) in the form of silicic acid (H4SiO4) to nodulated cowpea plants (Vignia unguiculata [L.] Walp.) grown in liquid culture resulted in considerable changes in the internal organization of nodule structure. Compared to the control plants which received no added silicate, bacteroid numbers increased significantly (P < or = 0.05) at silicate concentrations of both 0.096 and 0.48 g/l. The number of symbiosomes also increased by 3.2-fold at the silicate concentration of 0.96 g/l compared to the control. In contrast, the size of bacteroids and symbiosomes decreased significantly (P < 0.05) inside nodules of silicate-treated plants. The peribacteroid space was also decreased considerably (P < 0.05) with the application of 0.096 and 0.96 g of silicate per liter to plants. However, the size of intercellular spaces adjacent to infected and uninfected interstitial cells within the nodule medulla increased significantly (P < or = 0.05) at 0.096 g of silicate per liter followed by a sharply marked (P < or = 0.05) decrease with each subsequent increase in silicate application. The result was a large decrease (P < 0.05) in the area of bacteria-infected tissue occupied by intercellular space at the highest silicate concentration, which was caused by a significant (P < or = 0.05) increase in cell wall thickness. Our findings show that the positive effects of silicon on N2 fixation might actually be due to an increased number of bacteroids and symbiosomes. PMID- 11732203 TI - Ultimobranchial glands in the teleost (Plecoglossus altivelis ryukyuensis): special references to changes of gland volume with maturation. AB - In lower vertebrates, the bone-mineralizing hormone "calcitonin" is secreted from ultimobranchial glands, which assume various shapes, e.g., follicles, cellular masses, or cell strands. Histological observations support the view that in some teleosts, the glands increase in size when females maturate. We determined the exact volume of the gland in juveniles and adult males and females of a teleost, Ryukyuayu (Plecoglossus altivelis ryukyuensis). Furthermore, we examined plasma Ca, Na and K levels. In this species, the gland was fundamentally composed of a single follicle. The gland volume was converted to numerical data under a certain condition. It thus became clear that the value of the follicle wall was significantly increased only in maturing and mature females with high plasma Ca levels and that the value of the lumen did not show any statistically significant changes during growth and maturation. PMID- 11732204 TI - Morphometric analysis of the human trigeminal nerve. AB - We evaluated the components of nerve fibers of the motor and sensory roots of the trigeminal nerve by morphometric analysis. Trigeminal nerves were obtained from 5 cadavers (males, aged 67-95) were stained by Masson-Goldner-Goto method and examined under the microscope using a morphometric image-analyzer. The area and perimeter of axons were larger in the motor root than in the sensory root. The size distribution of axons was wider in the motor root than in the sensory root and the distribution pattern was unimodal type. These findings suggested that nerve fibers of the human trigeminal nerve gave similar findings to those of other peripheral nerves, regarding axonal size distribution and relative size of motor and sensory nerve axons. PMID- 11732205 TI - Morphometric nerve fiber analysis and aging process of the human inferior alveolar nerve. AB - We studied morphometric nerve fiber analysis and the aging process of the inferior alveolar nerve (IAN). Human IANs were resected at the mandibular foramen. The preparation of sections involved fixation, washing, dehydration, embedding, sectioning and staining as described in our previous reports. We estimated the total number of myelinated axons in the IAN being average 26,200, the transverse area of those being average 37.1 microns 2, the average perimeter of those being 23.0 microns, the average circularity ratio of those being 0.85, respectively. According to these results, the IAN did not demonstrate notable age relation change in any measured item. The IAN differed from not only our previous data on the human motor peripheral nerves but also those on the human sensory peripheral nerves. With regard to the aging process, the IAN indicated morphological features compared with other peripheral nerves. PMID- 11732206 TI - Morphometric analysis of the human tibial nerve and the ageing process. AB - We analysed numbers and sizes of the human tibial nerve branch innervating the soleus muscle. The material was taken from 13 cadavers aged from 67 to 98 years. A linear regression analysis disclosed a significant age-related decrease in the mean number per unit area and the mean transverse area of axons. Such decreases with age may indicate atrophy and loss of motoneurons. Our results could help in understanding the correlation between morphology and function during the ageing process. PMID- 11732207 TI - Bilateral investigation of the anatomical relationships of the external branch of the superior laryngeal nerve and superior thyroid artery, and also the recurrent laryngeal nerve and inferior thyroid artery. AB - The relationships of both the external branch of superior laryngeal nerve with the superior thyroidal artery and the recurrent laryngeal nerve with the inferior thyroidal artery were examined on the fixed cadavers of 4 adult women and 26 adult men. A total of 32 external branches of superior laryngeal nerve, 16 on each side, were dissected. When left and right sides were assessed altogether, 71.9% were medial to the artery while 28.1% were in between the branches. No nerve was found to be lateral to the artery. Thus the type in which the nerve is exposed to surgical trauma was found to be present in 28.1% of the cases. Only 12 of the cadavers could be assessed for symmetry and three fourths were found to have bilaterally symmetric relationship. A total of 52 Recurrent Laryngeal Nerves, 27 on the left and 25 on the right side were dissected. When bilateral symmetry was assessed in 21 cadavers, one third of the Recurrent Laryngeal Nerves were found to be bilaterally symmetrical. Inferior Thyroidal Artery was missing in 4 sides; bilaterally in one cadaver and unilaterally in 2 cadavers. Thus, artery-nerve relationship was assessed on 48 sides, 25 on the left and 23 on the right. On the right, 39.2% were in between the branches while 30.4% were anterior to the artery and 30.4% were posterior. On the left, 52% were in between the branches whereas 44% were posterior and 4% was anterior to the artery. Thus the position of the nerve in between the branches had the highest incidence while the anterior position had the lowest, the differences being statistically insignificant. PMID- 11732208 TI - Ramification pattern and topographical relationship between the portal and hepatic veins in the left anatomical lobe of the human liver. AB - Sixty-one human livers obtained from donated Japanese adult cadavers were dissected to reveal the ramification pattern of the portal and hepatic veins, and their topographical relationship in the left anatomical lobe. The segmental portal vein supplying S2 (P2) tended to form a single stem, whereas that of S3 (P3) was usually double. An intermediate branch between P2 and P3 was observed in 23.0% of livers. In spite of variation between livers, definite P2 and P3 were identified in 47 specimens. One tributary of the left hepatic vein (LHV) was usually present for drainage of S2, and two tributaries were present for S3 (sometimes also for S2 and/or S4). The latter two tributaries of the LHV and the two subsegmental branches of S3 showed three patterns of three-dimensional interdigitations. From these results, the portal vein system did not seem to have a two segmental composition (i.e., S2 and S3) in 23.0% of specimens, whereas the hepatic vein system did not have an intersegmental course in 23.4%. Thus, there were obvious limitations in using each system to determine the liver segment. Taking the overlapping cases into consideration, the left anatomical lobe of 41.0% of specimens did not seem to fit the definition of Couinaud's liver segment. In addition, four patterns of fissure vein (or scissural vein), > 5 mm in diameter at its terminal, were identified: (1) middle hepatic vein type (left median vein, 9.8%); (2) LHV type (left medial vein, 41.0%); (3) true fissure vein (3.3%); and (4) absent cases (45.9%). The former two types also suggested limitations of the hepatic vein system as an indicator of the segmental border. PMID- 11732209 TI - Proximal focal femoral deficiency, contralateral hip dysplasia in association with contralateral ulnar hypoplasia and clefthand: a case report and review of literatures of PFFD and/or FFU. AB - A 14 year old boy with proximal focal femoral deficiency (PFFD) on left side, contralateral hip dysplasia in association with ulnar hypoplasia and cleft hand was seen our clinic one year ago. From our research, despite it is atypical presentation, this case in a broad context conforms with the femoral-fibula-ulna complex (FFU). This boy was born to healthy and nonconsanguineous parents. Pregnancy and delivery were uncomplicated, and no history of prenatal teratogen exposure, any drug ingestion, infection, diabetes mellitus or other conditions that could be associated with malformation. The patient refused any sort of surgical treatment. PMID- 11732210 TI - Possible role of K-ras oncogene in mutagenic activity of ethylnitrosourea on lymphocyte hyperproliferation in rat colon. AB - N-ethyl-N-nitrosourea (ENU) is a potential carcinogenic agent that is commonly used in industry. Therefore the present study aimed to find out the possible effects of this agent on the rat digestive tract especially on the colon. We have studied the complementary mutation activity of the exon 2 of K-ras oncogene by ENU treatment in rat colonal tissue. While the two experimental group rats were injected once a week with 20 mg/kg and 300 mg/kg body weight-body weight with ENU (i.p.), the last experimental group was administered only with PEG and the control group animals received no treatment. Following 45 weeks, all animals were sacrificed and colonal tissues were obtained. Tissues were processed for light and electron microscopy and also for molecular biological analyses. While no colonal tumour development was observed in the control and in the PEG treated group, an extensive tumour development was seen in a wide range of tissues in the high dose ENU treated group. The light and electron microscopical examination of the rat colonal tissue revealed a lymphocyte hyperproliferation in the submucosal region, an increased number of polymorphonuclear leukocytes (PMLs) and occasional epithelial lesions. The mutation analyses of exon 2 of K-ras by HpaII demonstrated more than one recognition sites in the ENU treated group whereas there was only one enzyme cognate site in the control group. PMID- 11732211 TI - Rosuvastatin--'the super statin'. PMID- 11732212 TI - Detection of beta-thalassaemia trait: a study of fifty families. AB - BACKGROUND: This study was carried out to detect beta-thalassaemia trait in the siblings of beta thalassaemia major children by Hemoglobin (Hb) electrophoresis. METHODS: Subjects were divided into two groups. Group 1 was the patients group comprising 100-subjects siblings of beta-thalassaemia major children and Group 2 was the control group with 25-subjects. RESULTS: It was observed that 58% of the siblings of beta-thalassaemia major children had beta-thalassaemia trait. Even in the control group 2 subjects had beta thalassaemia trait, which shows that thalassaemia gene is very common in Hazara division of Pakistan. PMID- 11732213 TI - Cancer urinary bladder--5 year experience at Cenar, Quetta. AB - BACKGROUND: Purpose of this study was to see the incidence, age, sex, geographical distribution, symptoms, personal habits, signs, histo-pathology, early diagnosis and management of cases of Cancer Urinary Bladder (Ca UB) in the patients coming to CENAR, Quetta, Pakistan. METHODS: A retrospective study was conducted at CENAR for a period of 5 years from 1st Jan. 1993 to 31st Dec. 97, in which about 100 cases of cancer of urinary bladder were included, out of which 82 patients were male and 12 were females. RESULTS: During our 5-year period of study, 3571 new cases of cancer were registered at CENAR, out of which 100 (2.8% of total No. of cases) were of Ca UB. Hence 20 new cases of Ca UB per year were registered at CENAR. The maximum number of cases was registered in 1996. CONCLUSION: Our study concluded that Ca UB occurs more in male with a male female ratio of 4.5:1 and a high incidence after 40 years of age. No patient below 20 was reported. Histopathologically, Transitional Cell Carcinoma was dominating (75%). Other histological types seen were squamous cell carcinoma (4%), Adenocarcinoma (3%), UD (5%) and HPNA (10%). A considerable number of patients were using different preparations of tobacco (cigarette smoking (6%), Hubble Bubble (5%) and Niswar (Snuff) (12%). The patients were mainly treated with Radiotherapy, because at the time of reporting they were already in stage II or beyond (97%). Some patients were also treated by surgery such as TUR, partial or radical cystectomy. A few patients (6%) also received chemotherapy. PMID- 11732214 TI - Rational prescription of medicines--a study of indoor patients at a tertiary care hospital. AB - BACKGROUND: A study was conducted at Ayub Teaching Hospital, Abbottabad, a tertiary hospital, to know whether drugs are prescribed rationally keeping in mind indications, interactions, contraindications, proper dosage and economy of the patient. METHODS: The study was a descriptive case study. Records of 200 patients admitted in various units of the hospital were analysed by a panel of pharmacologists and conclusions drawn. RESULTS: It was found that only 52% of patients receive prescriptions that were rationale in all aspects. Where as in rest of the patients the rationale could be challenged. CONCLUSION: This study stresses the need for more concentrated and dedicated effort towards prescription of medicines. PMID- 11732215 TI - Prevalence and management of ovarian tumours in Women and Children Hospital Abbottabad. AB - BACKGROUND: The purpose of study was to compare world incidence of ovarian tumours whether benign or malignant with that of our unit. We have also discussed management of all these patients at our center. RESULTS: In our unit out of total 1400 admissions in 24 months, 26 had ovarian tumours, out of these 6 were malignant. The over all malignancy in our unit was 23% as compared to 15% of over all world incidence, 24 of these patients, were admitted through the OPD with advanced size of tumours. None of them previously had any check up at proper Gynaecological Centers. All these 24 were admitted to the ward, investigated, operated and after receiving histopathology reports were further managed accordingly. Two patients had torsion of ovarian tumour and were admitted through emergency and operated as emergency cases. In our study group the majority, 77% had benign ovarian tumours in which simple salpingoophorectomies was performed. PMID- 11732216 TI - Birth weight percentiles by gestational age: a hospital based study. AB - BACKGROUND: There are no present or old large population-based birth weight normograms available for Pakistani population. The aim of the study was to develop birth weight for gestational age normograms for singleton and twin births, based on perinatal data collected prospectively. METHODS: Birth weight percentiles by gestational age were determined in women delivering at Agha Khan University Hospital Karachi from January 1992 to December 1994. This was a prospective cohort study and included all women who delivered from 28 weeks to 44 weeks of gestation. Singleton as well as twin pregnancies were included but women having intrauterine or intrapartum deaths were excluded. RESULTS: Data of 4041 live births were collected from 4041 deliveries. Total infants were 4112, there being 71 sets of twins. The male babies were heavier than female babies at each gestational week. There was a preponderance of male infants for each week. Out of 4041 deliveries, 414 babies were born before 37 completed weeks; percentage of preterm birth is 10.24%. Males were more likely to be born preterm then were females, although females were more likely to be of low birth weight. Out of 414 preterm births, 281 were male and 183 were female. Out of them data of 4030 live births was compiled and analysed; percentiles were formed and compared with other studies. PMID- 11732217 TI - Delay in the diagnosis and prognosis of caecal carcinoma--a study of 20 cases. AB - BACKGROUND: Caecum is the dilated part of the right colon situated in the right iliac fossa, therefore the etiology of this cancer is similar to those of the rest of the colon. Caecal carcinoma is more common in developed countries but it is not a rare disease in underdeveloped countries. This is more common in high socio-economic people who use less fibrous and purified diet. With improvement in health education and social status of the people of Pakistan, there is an emerging trend of Westernization in our society. This may increase the incidence of cancer in Pakistan. Carcinoma of caecum is curable disease is diagnosed early and treated. If we are aware of the pathogenesis, etiology, clinical presentation and management of the disease, we can offer a lot to these patients by diagnosing the caecal carcinoma at an earlier stage, this was the aim of this study. METHODS: 20 cases (15 males and 5 females) of Carcinoma of Caecum were studied from January 1997 to December 1999. Out of these six patients presented in emergency ward, 2 as acute appendicitis and 4 as intestinal obstruction. After routine non-specific investigations, our specific investigations included USG abdomen, barium contrast studies and colonoscopy, Barium study and USG remained of primary importance. RESULTS: Most of the patients were between age 45-65 years, oldest patient being 80 years old and the youngest was of only 30 years. Most of the patients presented as mass in the right iliac fossa and only some patients as intestinal obstruction. Right hemicolectomy with primary ileo transverse anastomosis was our standard surgical procedure and was done in about 14 cases. But in some cases ileo-transverse by-pass operation was done as the growth was locally advanced. However, only the omental biopsy was taken in the cases where carcinoma was too advanced. CONCLUSIONS: We think that our sociodemographic factors put a hindrance in detection of this carcinoma at early stages and therefore curative procedures are difficult to bear good results. PMID- 11732218 TI - Intraocular pressure changes after Nd-YAG laser capsulotomy. AB - BACKGROUND: 28-43% of the patients develop posterior capsular thickening after extra capsular cataract surgery. Nd-YAG laser capsulatomy is a method of choice to treat this complication. METHOD: We conducted a prospective clinical, randomized comparative study to evaluate the post YAG laser IOP rise in 54 patients at Ayub Teaching Hospital Complex. RESULT: There were 43% male and 57% female patients with a mean age of 63.6 years. The mean time for capsular opacification was 14.6 months. Postoperative visual improvement was 6/24 to 6/6. All patients showed post laser IOP rise that was controlled by Topical Beta blockers and steroids effectively. PMID- 11732219 TI - Biliary Ascariasis in children. AB - BACKGROUND: Ascariasis is very common in this part of the world. Biliary Ascariasis is rare but is the commonest extra-intestinal complication. METHODS: This study was conducted At Ayub Teaching Hospital, Abbottabad, a tertiary care hospital. Five cases of biliary Ascariasis were studied from December 1999 to January 2001. History of passage of worms in stool or vomiting and abdominal pain was taken. After clinical evaluation, Stool Examination, Serum amylase, LFTs, and complete blood picture (CP) were done. Ultrasound abdomen was done as a basic tool for diagnosis. All cases were given Piperazine. Spasmolytics were also given to relax sphincter of oddi to release the worms. Surgical opinions were also taken for each case. RESULTS: About 80% Children were above 10 years of age. One case was 1 1/2 years old. Most common symptom was right upper quadrant abdominal pain. Previous history of passage of worms in stool and vomitus was also present. Two children developed complications of Biliary Ascariasis i.e., Cholecystitis and Portal empyema and responded to antibiotics. All children became symptom free in about 48 hours of treatment. Ultrasound was found to be a reliable, non invasive, and quick tool for diagnosis and follow up. PMID- 11732220 TI - Lactation, breast-feeding supplementation and nutrition. AB - BACKGROUND: Breast-feeding and proper weaning methods are very important to cover the milestones of life and to achieve good nutritional status under five. The proper lactation, feeding and supplementation methods must be adopted to achieve this goal. In Pakistan a lot of agencies have been working on this. It is included in the daily effort of every doctor and medic to achieve this goal. METHODS: This study was done to know the results of our efforts in Oghi Pakistan. A Hospital based study on 10 children/mothers was conducted. The children were randomly selected from the best E.P.I. center of Oghi. The mothers were interviewed after explaining them the purpose of the study. The questionnaire was filled and questions were explained in local language for the convenience of mother. The parameters studied and questions asked included age, sex, weight, height, first feed, age at weaning, type of milk, and type of weaning food. RESULTS: The results were found satisfactory due to the hard work of workers of UNICEF, Health department of NWFP and better when compared to other areas of the province. PMID- 11732221 TI - Effect of maternal anaemia on fetal parameters. AB - BACKGROUND: A cohort study of anaemic pregnant women was carried out at the Department of Obstetrics and Gynecology, Women and Children Hospital, Abbottabad, from November 30, 1992 to March 30, 1993, to determine the effects of antenatal maternal anaemia on the newborn babies. METHODS: 34 pregnant women with anaemia and 39 non-anaemic pregnant women were included in the study. Parameters studied were Hemoglobin levels, RBC morphology, Foetal birth weights, Foetal gestational ages, and Foetal APGAR score (at birth and after 10 months). RESULTS: Out of 34 anaemic women, 6 delivered at 7 or 7+ months, 7 at 8 or 8+ months and 21 at 9 or 9+ months, as compared to 39 non-anaemic, one of whom delivered at 7 or 7+ months, 2 at 8 or 8+ months and 36 at 9 or 9+ months. Fetal mortality rate for anaemic women was 50% at 7 or 7+ months, 28.7% at 8 or 8+ months and 23.80% 9 or +9 months. For all non-anaemic women, it was 0%. Mean fetal birth weight in anaemic group was 1.4 +/- 0.66 kg at 7 or 7+ months, 1.93 +/- 0.53 kg at 8 or 8+ months and 2.61 +/- 0.31 kg at 9 or 9+ months. Among non-anaemic group, it was 2.8 kg at 7+ months, 3.75 +/- 0.55 kg at 8 or 8+ months and 3.65 +/- 0.66 kg at 9 or 9+ months. CONCLUSIONS: These findings show that mothers with nutritional or iron deficiency anaemias tend to deliver prematurely with low birth weight babies and a high mortality rate or stillbirths, as compared to non-anaemic mothers. PMID- 11732222 TI - FBG and TC/HDL ratios in type 2 diabetes mellitus. AB - BACKGROUND: Presence of dyslipidemia, i.e. raised Total Cholesterol (TC) and decreased High Density Lipoproteins (HDL) is an established phenomenon in type 2 diabetes mellitus. Its relationship to the fasting blood sugar (FBG) level in both diabetics and non-diabetics has yet to be established. The present study aims to show such a relationship. METHODS: 150 subjects were selected and divided into two groups, one of 50 controls (non-diabetics), the other of 100 patients with type 2 diabetes mellitus. Their FBG levels were determined by enzyme oxidase method, and their TC and HDL levels were determined using a standard kit method. RESULTS: FBG and TC:HDL ratios were increased in the patient group (p = < 0.001). Comparison of FBG and TC:HDL ratios revealed a highly significant rise (p = < 0.001) in the patient group. FBG with TC:HDL ratios between both groups showed a positive correlation (r = 0.554). CONCLUSION: The results of the present study suggest an association between FBG and TC:HDL ratios, which may be a contributory factor to the increased prevalence of coronary artery disease in patients with type 2 diabetes mellitus. PMID- 11732223 TI - Heterotopic pregnancy--a report of 2 cases in women without risk factors. AB - Two cases of heterotopic pregnancy are reported. The cases emphasize that co incident pregnancies may occur in women who are without risk of ectopic pregnancy or multiple gestation. The cases describe the complexities of diagnosis and management. PMID- 11732224 TI - Liver function tests in patients of pulmonary tuberculosis using four different drug regimens. AB - BACKGROUND: Chemotherapy is the basic approach to clinical tuberculosis control. Antituberculous therapy causes derangement of hepatic functions revealed by disturbed liver function tests. The incidence of side effects may vary depending upon a number of factors. The primary purpose of this study was to determine the relative and absolute hepatotoxicity of different antituberculous drug regimens in Pakistani population where majority of the tuberculous patients belong to poor socioeconomic status. METHODS: One hundred patients between 30 to 70 years of age with newly diagnosed pulmonary tuberculosis were selected and divided into four groups on the basis of different drug regimens. Blood and urine tests of these patients were made. Liver function tests were performed before therapy and then after one, two, three, six and nine months of treatment. RESULTS & CONCLUSIONS: Antituberculous therapy causes derangement of hepatic function to a variable extent in patients of four different antituberculous drug regimens under study. Drug combination of Streptomycin, Isoniazid and Myambutol seem to be best tolerated of all the four regimens. Monitoring of the liver function in patients on antituberculous therapy indicated that liver dysfunction most frequently occurs during first three months of therapy. There is a tendency for enzyme values to return to normal inspite of continuous treatment. The mechanism underlying this adaptation to injury to the liver is unknown. Biochemical tests in the patients presenting with jaundice yielded the pattern of acute hepatocellular necrosis with high transaminases and moderately elevated Alkaline Phosphatase. None of the patients had hepatitis associated antigen in their serum. PMID- 11732225 TI - [Osteoporosis and hormonal contraception]. AB - OBJECTIVE: To analyse the influence of the contraceptive treatment on bone mass. DESIGN: Literary review. SETTING: Department of Gynaecology and Obstetrics, 1st Faculty of Medicine, Charles University, Prague, Czech Republic. METHOD: Informations were collected from full-texts which were taken from database Medline. RESULTS AND CONCLUSION: The lowering of bone turnover under combined oral contraception was shown in all studies. It is favourable for bone mineral density after 30 years when bone loss starts. The positive influence is significantly dependent on the length of treatment. Only gestagen drugs have no positive effect on bone mass. The influence of contraception on mineral density in term of physiological bone density increasing before 20-25 year is not clear. Some of studies informed us about adverse effects of extremely-low dose oral contraception on mineral density in this group. PMID- 11732226 TI - [Polymorphism of angiotensin converting enzyme (ACE) and TNF-beta genes in men with disorders of spermatogenesis--pilot study]. AB - INTRODUCTION: The presence of components of the renin angiotensin system and tumour necrosis factor in a male reproductive tract supports the hypothesis that these substances may influence reproductive functions. It was proved that angiotensin II as a product of ACE has influence on sperm capacitation and motility. TNF-beta is released from T-lymphocytes and has the regulatory effect on steroidogenesis and spermatogenesis. Aberrations of these agents can result in infertility. OBJECTIVE: The aim of this study was to determine the allele frequency of ACE and TNF-beta genes in men with pathological sperm count and men with normal fertility. We examined the insertion/deletion (I/D) ACE and B1/B2 TNF beta gene polymorphic alleles and analyzed their frequency in patients and fertile men. DESIGN: A pilot study. SETTING: 1st Clinic of Gynaecology and Obstetrics and Institute of Pathologic Physiology, Masaryk University, Brno, CR. MATERIAL AND METHODS: The genomic DNA was isolated from peripheral blood leukocytes by a standard method according to Sambrook in a group of 46 patients (33.4 +/- 7.2 years) with pathological sperm count (9 azoospermia, 21 severe oligoasthenospermia, 16 moderate oligoasthenospermia) and in a control group of 88 healthy men (31.2 +/- 9.3 years) with normal fertility. Polymerase chain reaction (PCR) was used for genom analysis. The method according to Rigat was used for the I/D ACE polymorphism. B1/B2 TNF-beta genotype of each patient was determined after Nco I digestion of the amplified product and subsequent agrose gel electrophoresis. Fisher's exact test and chi square test were used for statistical analysis. RESULTS: In the study we found these differences of allel frequency and their combination: 1. Combinations of the genotype II (ACE) + B1B2 (TNF-beta) and genotype II (ACE) + B2B2 (TNF-beta) were less frequent in patients (8.7%) than in fertile men (28.4%), this difference was statistically significant (p = 0.021). 2. Allele B1 (TNF-beta) was more frequent among patients (40.2%) than in the control group (29.5%), this difference was near to the point of statistical significance (p = 0.05). 3. Allele D (ACE) frequency was higher in men with pathological sperm count (52.2%) than in fertile men (44.9%), this difference was not statistically significant (p = 0.15). CONCLUSION: The study has found different allele frequency of I/D ACE and B1/B2 TNF-beta genes polymorphism in men with pathological sperm count compared to men with normal fertility. These results could contribute to elucidate the genetic background of a male infertility. PMID- 11732227 TI - [A shortened exposure of oocytes to sperm in standard in vitro fertilization]. AB - OBJECTIVE: To find out if the shortened exposure of oocytes to sperm can influence the embryo quality and the implantation and pregnancy rate of the classical IVF also in Centre for Assisted Reproduction. DESIGN: A prospective study including 104 patients undergoing the classical IVF procedure. SETTING: Centre for Assisted Reproduction, Clinic of Gynaecology and Obstetrics, Palacky University Medical School and the University Hospital in Olomouc. METHODS: In the study group with the shortened exposure (54 patients) the oocytes were washed one hour after the insemination and transferred into the fresh medium. In the control group (50 patients) the classical procedure (e.g. transfer into the fresh medium 17 to 20 hours after insemination) has been used. The following parameters have been evaluated in both groups: the stimulation protocol, sterility cause, number of previous IVF cycles, age, serum estradiol level on the day of hCG application, number of retrieved oocytes, fertility rate (FR), embryo quality, pregnancy rate (PR) and the implantation rate (IR) expressed as the number of gestational sacs with heart action related to the number of transferred embryos. T-Test, Mann Whitney Test and chi 2 were used for statistical evaluation. RESULTS: There were no statistically significant differences between the study and the control group in any of the parameters compared. CONCLUSION: In our study group the shortened exposure of oocytes to sperm did not influence the results of the classical IVF. PMID- 11732228 TI - [Evaluation of success in assisted reproduction]. AB - OBJECTIVE: Summarize different ways of assisted reproduction performance evaluation. DESIGN: Review. SETTING: Centre for Assisted Reproduction, Department of Gynaecology and Obstetrics, 1st Faculty of Medicine, Charles University, Prague. METHODS: In databases of assisted reproduction centres, in national and international registers we evaluate data concerning in vitro fertilisation and related methods. Target of this evaluation is, first of all, to increase efficiency. RESULTS: Outputs of all the registers inform about absolute numbers of cycles, deliveries, born babies and about relative parameters (implantation rate, pregnancy rate), describing the efficiency of the treatment. CONCLUSION: The output evaluation must consider the influence of the age of female, number of transferred embryos and other factors. PMID- 11732229 TI - [DNA analysis of chromosome Y in the area of the azoospermia factor (AZF) in infertile men]. AB - OBJECTIVE: Establishment of investigation of sterile male DNA in AZF region- choice of loci and primers for investigation, optimization of PCR conditions (polymerase chain reaction). DESIGN: For practice. SETTING: Department of Medical Genetics and Foetal Medicine, Faculty of Medicine, Palacky University and Faculty Hospital Olomouc. METHODS: PCR amplification of DNA isolated from blood of sterile men and consequential electrophoresis of synthesized DNA fragments to appoint microdeletions in AZF. RESULTS: From January to June 2000 were detected the microdeletions in AZF of 3 out of 79 sterile men (3.80%) by means of the Experteam firm kit. From July to December 2000 were tested and optimized conditions of amplification of 10 AZF loci to substitute the kit and they were used for examination of the first 20 sterile men of our collection. CONCLUSION: In our laboratory was established routine examination male sterility related to microdeletions in AZF. With our collection of loci was substituted the original Experteam firm kit and was widened spectrum of observed loci. PMID- 11732230 TI - [Ascitic fluid puncture in the treatment of severe forms of ovarian hyperstimulation syndrome]. AB - OBJECTIVE: Evaluate the contribution and complications of ascitic fluid puncture in the treatment of severe ovarian hyperstimulation syndrome. DESIGN: Retrospective analysis of 18 patients. SETTING: Department of Gynaecology and Obstetrics, University Hospital, Palacky University, Olomouc. METHODS: 18 patients with the diagnosis of severe ovarian hyperstimulation syndrome admitted to intensive care unit from January 1996 to June 2000 were analysed. As a part of the therapy of severe OHSS 11 punctures of ascitic fluid was performed. In 3 patients the reinfusion of the ascites into the blood circulation was performed. RESULTS: The transabdominal paracentesis under the ultrasound control in 11 patients was performed. In 6 patients the puncture had to be repeated. The amount of removed ascitic fluid in single procedure was between 600-3750 ml. Total amount of fluid in different patients was between 1500-17,300 ml. Sterility of ascites and high contents of proteins was proved. In 3 patients with the refractory ascites the reinfusion of the ascitic fluid to the circulation was performed. No serious complications of the punctures were observed, 4 patients had a slight rise of body temperature, which fell down without antibiotics therapy. CONCLUSION: Puncture of the ascitic fluid is a save and effective part of the therapy of severe OHSS. In the treatment of the refractory ascites the reinfusion of the ascitic fluid should be used. PMID- 11732231 TI - [Ethical problems in human embryo research]. AB - The problem of human embryo research is closely connected with the moral status of preembryo, embryo and fetus. Nevertheless the moral consensus on the status of this human entities is not feasible. Research activities in this fields are divided into two subcategories: In vitro research and research on implanted fetuses. Both needs to clarify one significant ethical distinction: it is the situation when the embryo is planned to be born, or to be perished. It is concluded, that the public discussion on this topic is necessary. PMID- 11732232 TI - [Magnetic resonance in urogynecology]. AB - OBJECTIVE: The aim of the study was to review the use of the MRI in urogynaecology and to describe an original method for the construction of 3D computer models of pelvic floor. DESIGN: Pilot study. SETTING: Department of Obstetrics and Gynaecology, Institute of Sexuology, 1st Faculty of Medicine, Charles University, Prague, Department of Radiodiagnostics, Charles University, Hradec Kralove. METHODS: A review of the use of the MRI in urogynaecology was made. A new method was introduced for creation of computer 3D models of female pelvis based on the data obtained by the MRI. RESULTS: Five complex computer models of the female pelvic organs and pelvic floor were created. Many so far estimated details were visualized. CONCLUSION: The use of MRI in urogynaecology can bring important data about the functional anatomy of female pelvic floor. PMID- 11732233 TI - [Placental and umbilical cord changes in drug-addicted women]. AB - OBJECTIVE: The aim of the study was to gauge the impact of drugs on placental changes in heroin and pervitin addict pregnant women as one of the influence factors supporting the lower birth-weight of the neonate. DESIGN OF THE STUDY: Prospective study. SETTING: The Department of Gynaecology and Obstetrics and the Department of Pathology of the General Teaching Hospital and the 1st Medical Faculty of Charles University in Prague. METHODS: The drugs-addict pregnant women were included in the study in the period between January 1998 till the end of the year 2000. We succeeded to concentrate in total 39 drug abused pregnant women and 39 their placentas and umbilical cords were examined (19 heroin and 20 pervitin addict). The placentas were sent for the histological examination in total without the chemical fixation. There were 3 placental samples, 2 cord and 2 membranes samples examined. The controls groups were formed by non-abused pregnant women and their placentas by method of accidental choice. RESULTS: We found the statistically significant higher incidence of placental and cords abnormalities in among the addict women compared to their non abused controls (P < 0.05). In heroin addict group in 16 from 19 cases were the abnormalities present. We registrated increased intervillous and perivillous microfibrin deposits (6 times), an increased number of trophoblastic proliferation buds (7 times) increased vascularization of the villi (7 times) mikrovilli (6 times) and their necrosis (5 times). In umbilical cord we enregistered in 3 cases the only one artery, in two case the thrombosis of the vein. In 5 cases we also found the signs of infection. In pervitin addict group we registered placental changes in 9 from 20 cases. The majority of them presented as increased microfibrin deposits on the surface and also in the choriotic board (9 times), in five cases we enregistered the intervillous haemorrhage manifested in 2 cases clinically as abruption of placenta. The placental changes participate in low birth weight of neonates, IUGR and the abbreviation of the gestational duration. CONCLUSION: The microscopic changes in placenta is the expression of the circulatory disorder during the attack of the drug. They are not four-square specific, rather quantitatively expressed. In heroin abused women is the incidence of placental changes much more higher comparing to pervitin-addict. Heroin led more frequently to preterm labour, lower birth-weight and IUGR. The pervitin-addict mothers are higher risk of placental abruption. PMID- 11732234 TI - [Epidural obstetrical analgesia and cardiotocography]. AB - OBJECTIVE: The increase of cardiotocographic abnormalities, such as late decelerations, bradycardia and reduced variability have been reported with epidural analgesia (EDA) in some studies. The aim of our study was to assess the influence of epidural analgesia on the incidence of the pathological cardiotocographic patterns. DESIGN: A clinical study. SETTING: Department of Obstetrics and Gynaecology, Jessenius Medical Faculty of Comenius University, Martin, Slovak Republic. METHODS: Clinical group consists of 200 parturients, who borne in our department from 1997 to 2000. The epidural group of parturients (n = 100) borne under epidural analgesia, the control group of (n = 100) borne under other resp. no analgesic method. We compared BFHR, variability and the incidence of accelerations and decelerations in CTG between both groups. All cardiotocographic patterns were evaluated by Fischer score. For statistic evaluation was used Student t-test. RESULTS: We did not find any statistically significant differences in Fischer score between epidural and control group. CONCLUSION: The epidural labour analgesia is safe method of ease pain during labour. In our study we confirmed, that EDA is not responsible for the increase of pathological cardiotocographic patterns. PMID- 11732235 TI - [Ectopic omental pregnancy--laparoscopic diagnosis and therapy]. AB - OBJECTIVE: Description of a very rare case of primary omental pregnancy. DESIGN: Case report. SETTING: Institute for the Care of Mother and Child, Prenatal Diagnostic Centre, Prague, Institute for Postgraduate Medical Education, Prague. SUBJECT AND METHOD: Implanted product of spontaneous conception was found in the omentum. The gestational sac on omentum had the appearance of a haemorrhagic tumor 2 cm in diameter. CONCLUSION: After dissection, an intact embryo of 7 mm with a 4 mm yolk sac was discovered. PMID- 11732236 TI - [Preoperative ultrasound evaluation of myometrial invasion in patients with endometrial carcinoma]. AB - OBJECTIVE: To evaluate the diagnostic accuracy of preoperative transvaginal sonography in the detection of myometrial invasion in endometrial cancer patients classified by the grade of disease. DESIGN: Prospective multicenter study. SETTING: Department of Obstetrics and Gynaecology, Medical Faculty, Palacky University, Olomouc, Czech Republic. METHODS: In the period 1998-2001, 195 patients from 10 cooperating centers in the Czech Republic were enrolled in the RHOCE (risk of hysteroscopy in endometrial cancer) study. Sixteen patients did not meet the vaginal sonography study criteria (insufficient surgical staging, missing ultrasound examination). Sonographic results in 179 patients were categorized as M0--absence of infiltration (n = 97), M1--infiltration less than 1/2 myometrial depth (n = 61), M2--infiltration greater than 1/2 myometrial depth (n = 21). The final histopathology report of myoinvasion was used as a golden standard. RESULTS: We proved lower accuracy of vaginal sonography in detecting myometrial invasion in patients with higher grade of disease. CONCLUSION: Based on these data, the value of preoperative vaginal sonography as the sole criterion in the decision to perform extensive surgical procedures is questionable and warrants further evaluation. PMID- 11732237 TI - [Importance of hepatitis C virus infection in gynecology and obstetrics]. AB - OBJECTIVE: Diagnostic methods of hepatitis C-virus. SUBJECT: A review article. SETTING: 1st Internal Clinic, General Faculty Hospital in Prague, 1st Medical Faculty, Charles University. SUBJECT AND METHOD: Hepatitis C virus infection importance is increasing during the last few years, in concordance with this fact therapeutical options are being developed very rapidly, too. Majority of cases of HCV infection has a silent clinical course for a long time and patients are often diagnosed in the late stage of liver disease. This is the reason why the diagnosis should be established as early as possible. Gynecologist can be the first person who can observe the symptoms of the infection and therefore he may become the key person in the diagnostic process. CONCLUSION: The basic facts about the diagnostic methods and clinical course of the disease are mentioned in this review. The differences in clinical course of HCV infection in pregnancy and the vertical transmission is discussed in details. PMID- 11732238 TI - [Hormonal contraception and thromboembolic disease--pathophysiologic findings and practical recommendations]. AB - OBJECTIVE: Review the knowledge about changes in hemostatic system in combined oral contraceptives users (focused on studies published between 1997 and 2000). TYPE OF STUDY: Review of literature. SETTING: Department of Clinical Hematology, University Hospital Hradec Kralove, Czech Republic. METHODS: Summary of the results of substantial studies published on this topic. Studies were identified by Medline database search. RESULTS: Hemostatic changes were described in all subsystems (in both procoagulant and fibrinolytic systems, in natural inhibitors of coagulation) and could be seen also in molecular marks of coagulation and fibrinolysis activity. Substantial differences were not described for ethinylestradiol doses 20-50 micrograms and up to 1997 also not for different progestins. More expressed acquired resistance to activated protein C at 3rd generation progestins in comparison with levonorgestrel was described recently as well as greater inhibition of fibrinolysis at desogestrel users. CONCLUSION: The results of recent studies indicate the differences in hemostatic changes between users of 3rd generation progestins, respective desogestrel, and users of levonorgestrel. However, the practical recommendations (aiming at minimalization of VTE risk) are focused on personal and family history and on patient counseling. The choice of progestin could be important but it is not crucial. PMID- 11732239 TI - [Meconium and postnatal neurologic handicaps]. AB - OBJECTIVE: A review of meconium pathophysiology and its contribution to the incidence of postnatal neurological handicap. DESIGN: Reviewed article. SETTING: Department of Gynaecology and Obstetrics, Charles University and Faculty Hospital Plzen, Czech Republic. SUBJECT AND METHOD: Meconium can be a cause of infant neurological handicap. Two main pathogenetic pathways are mentioned. 1. Meconium (and its components: bile acids) may have a direct vasoconstrictive effect on umbilical and placental vessels. This way still remains controversial. 2. Meconium as a possible cause of intraamniotic infection results in a release of fetal cytokines (TNF alpha, IL-1 beta, IL-6), which can damage myelinogenesis in periventricular white matter. RESULTS: Meconium in premature labour is a higher risk factor compared to term delivery. 41% of premature infants were diagnosed as having CP when meconium was present compared to 10% in the same group with clear amniotic fluid. The incidence in term pregnancy with meconium present is 0.4% compared to 0.3% in a population without any obstetrical risk. CONCLUSION: Ultrasonographically found periventricular leukomalacia is the most reliable sign of subsequent cerebral palsy or other neurological sequelae. PMID- 11732240 TI - [Postcoital contraception]. PMID- 11732241 TI - [Recommendations for prescribing postcoital contraceptives]. PMID- 11732242 TI - Drinking patterns, health care utilization, and costs among HMO primary care patients. AB - A survey of 8,034 primary care patients in a health maintenance organization examined the relationship between alcohol consumption and health care costs and service use. Costs were estimated from service use data for 1 year before and 2 years after study enrollment. No strong, consistent relationships were identified between multiple indicators of drinking patterns and either health care costs or service use. Compared with total costs among very light drinkers, former drinkers were higher, lifetime abstainers were similar, and persons in the higher drinking levels tended to have lower but not significantly different costs. Drinking patterns did not appear to be an important predictor of short-term health care costs or service use in this setting. Further study of former drinkers is warranted to examine the role of alcohol-related illnesses in the decision to quit drinking. PMID- 11732243 TI - A confirmatory factor analysis of the BASIS-32 in racial and ethnic samples. AB - The present study performed confirmatory factor analysis across major racial and ethnic groups of the BASIS-32, a measure of functional status of persons receiving mental health treatment and suitable for routine assessment mental health care. The purpose was to perform a preliminary investigation of cross cultural equivalence in a county-level mental health program in a major metropolitan area. The results indicated a factor structure similar to that reported in the literature, and they suggested acceptable levels of agreement in structure between racial and ethnic minority groups and whites. The study revealed little reason to believe that the BASIS-32 varied in underlying structure across racial and ethnic boundaries, although further research is indicated. PMID- 11732244 TI - An analysis of stressors and co-morbid mental health problems that contribute to youth's paths to substance-specific services. AB - Substance-specific services are tailored to address the inappropriate use of chemicals such as alcohol and marijuana. Unfortunately, few teens ever access such need-based services. This article explores the paths to substance-specific service use in a sample of urban adolescents who are public service sector users: a direct path from substance misuse and an indirect path from general mental health service use. Results indicate that even though a significant percentage of youths frequently consume substances and display negative use-related behaviors, few actually received substance-specific services. Contributors to the substance misuse path included youths' family substance dependence and environmental stressors. Contributors to the mental health services path were comorbid depression and substance misuse. PMID- 11732245 TI - The involvement of a consumer council in organizational decision making in a public psychiatric hospital. AB - This article describes a consumer group within a public psychiatric hospital that serves primarily a forensic population. Some barriers to participation included the severity of some clients' mental illness, an organizational culture that does not fully support participation, the lack of clients' awareness of problems or alternative actions, and inherent power imbalances between clients and staff. Despite these barriers, the consumer group has made improvements for facility clients. Some factors associated with this success included strong administrative support, the allocation of a highly qualified staff liaison to work with the group, and the integration of the group into the facility's formal decision making structure. Lessons are offered for the development of similar groups within public psychiatric hospitals and community-based mental health agencies. PMID- 11732247 TI - The accuracy of medical record documentation in schizophrenia. AB - Medical records are commonly used to measure quality of care. However, little is known about how accurately they reflect patients' clinical condition. Even less is understood about what influences the accuracy of provider's documentation and whether patient characteristics impact documentation habits. Discrepancies between symptoms and side effects evaluated by direct assessment and medical records were examined for 224 patients with schizophrenia at two public mental health clinics. Multivariate regression was used to study the relationship between patient, provider, and treatment characteristics and documentation accuracy. Overall, documentation of symptoms and side effects was frequently absent. Documentation varied substantially between clinics, and it was generally less likely for patients who were severely ill, black, or perceived as noncompliant. The accuracy and consistency of medical record documentation should be demonstrated before using it to evaluate care at public mental health clinics. PMID- 11732246 TI - Does managed mental health care reallocate resources to those with greater need for services? AB - Evidence points to the existence of two coexisting inefficiencies in mental health care resource allocation: those with need receive too limited or no care while those with no apparent need receive services. In addition to reducing costs, managed mental health care is expected to reallocate treatment resources to those with greater need for services. However, there are no empirical findings regarding this issue. This study tests whether managed mental health care has had a differential impact by level of need. Data consist of three waves of a community sample with a control group. The study finds that managed care has not succeeded in reallocating resources from the unlikely to the definite "needers." PMID- 11732248 TI - Using a computerized patient database to evaluate guideline adherence and measure patterns of care for major depression. AB - This study examined the translation of recommendations from the Agency for Health Care Policy and Research (AHCPR) guidelines for major depression into measures derived from a computerized database to assess guideline conformance and patterns of care for major depression. Patients (n = 208) were identified who were hospitalized for major depression and had two or more outpatient mental health appointments within 6 months of discharge from an academically affiliated Veterans Affairs Medical Center. Measures were based on AHCPR guideline recommendations or developed independently. Conformance could be measured for three guideline recommendations. Of patients on single-agent antidepressant therapy, 87% received dosages within the recommended range. Sixty-nine percent received the recommended number of follow-up visits. Specific condition-related treatment interventions were identified in 32% of patients with concurrent alcoholism. Dual diagnoses of depression and drug or alcohol abuse were not deterrents to prescribing benzodiazepines. Despite its limitations, computerized database analyses provided efficient measures of guideline adherence. PMID- 11732249 TI - Understanding urban child mental health l service use: two studies of child, family, and environmental correlates. AB - The results of two studies identifying child, family, and environmental correlates of initial and ongoing mental health service use by urban minority children and their families are presented. In the first study, data from a sample of 405 adult caregivers of children revealed no predictive power of child demographic characteristics in relation to initial or ongoing service usage. Only parental ratings of child impulsive-hyperactive behavior were significantly related to ongoing involvement in services. In the second study, a new sample of 100 urban caregivers of children was interviewed. Parental discipline efficacy and attitudes about mental health services were found to relate significantly to initial attendance. Relative to ongoing service use, level of family stress, presence of another adult in the home, and parental discipline efficacy were significant. Implications for research and child mental health service organizations are highlighted. PMID- 11732250 TI - Raising our sites: dissemination of supported education. AB - In order to promote replication of supported education, an exemplary rehabilitation model for adults with psychiatric disabilities, funds were accessed through a Community Action Grant from the Center for Mental Health Services of the Substance Abuse and Mental Health Services Administration. Three communities in Michigan participated in a multistage process designed to maximize community ownership by encouraging local adaptations involving all stakeholder groups and providing technical assistance. The stages in the process were organizing the community for supported education development, acquiring knowledge about supported education basics, collecting information (needs assessment and barrier identification), and developing the plan. All three sites have begun implementation, providing services to adults with psychiatric disabilities who wish to pursue post-secondary education. The approach employed has applicability for other local communities. PMID- 11732251 TI - Comment on "Managed care in the public sector: lessons learned from the Los Angeles PARTNERS program". PMID- 11732252 TI - [Acetabular roof remodeling after varus derotation osteotomy in congenital hip dysplasia]. AB - The aim of this paper was to evaluate the acetabular roof remodelling process and changes of the Wiberg angle after proximal femur varus drotational osteotomy up to skeletal maturity. The analysis included 33 patients after 48 osteotomies (in 15 patients on both sides). The age at the time of surgery ranged from 1.0 to 9.8 years, mean 4.7 years. Follow-up time ranged from 7.6 to 15.9 years, mean 12.0 years. Statistical analysis showed significant differences in acetabular index, Sharp's angle, acetabular ratio and Wiberg's angle between early post-operative x rays and radiograms taken at the last follow-up. Comparison not only of the remodelling dynamics, but also final radiological measurements confirm the necessity for surgical of severely dysplastic acetabular roof, especially in older age groups. PMID- 11732253 TI - [The role of cytokines in aseptic loosening of total hip prosthesis]. AB - The aim of this study was estimation of the secretory activity of macrophages isolated from granulomatotic tissue in aseptic total hip prosthesis loosening (particularly in regard to IL-6, TNF-alpha and OSM secretion). A considerable increase of IL-6 and TNF-alpha (proosteolytic cytokines) content was noted. At the same time a prominent decrease in OSM (an osteoprotective cytokine) was noted. The obtained results seem to suggest that the changed immunobiology of macrophages consisting of increase osteolytic activity and decreased osteoprotective abilities could be responsible for the loosening of hip prosthesis. PMID- 11732254 TI - [Early results of high tibial osteotomy with the DERO system for osteoarthritis of the knee]. AB - The authors present the surgical technique and early results after 14 wedge "minus" osteotomies and 5 wedge "plus" osteotomies performed in 16 (age ranging from 45 to 71 years) patients with osteoarthritis of the knee. Use of the DERO system for external stabilization of the knee allows early mobilization of the joint. According to the HSS scale in 15 cases (18 knees) a good result was achieved. Self-assessment of the results of treatment by the patients w?yielded good results in 12 cases and satisfactory in 3. All patients report greater ease of ambulation on flat terrain, across stairs, an increase in ROM and a decrease of pain symptoms. In one case a bad result was caused by infection of the surgical wound. In the observed group no infections, no reaction d?around the pins nor loosening of the bars was noted. PMID- 11732255 TI - [The role of risk factors in exploitation of lower extremity prosthesis after amputation in adults]. AB - The author presents the influence of gender, age, main disease that lead to amputation, other concomitant diseases, side of amputation and number of years since amputation on prosthesis exploitation by adult patients. The aim of this paper was to assess which risk factors and to what degree influence the exploitation of prosthesis of the lower extremities. The analysis was based on the Yule Q correlation factor. The best prognosis was found for male patients less than 60 years old with below-the-knee traumatic amputations. The worst prognosis was for female patients with amputations at femur level with concomitant diabetes and smoking. PMID- 11732256 TI - [Residual clubfoot correction by means of lateral cuboid resection]. AB - The paper presents the late results (after 5-16 years; mean follow-up--9.5 years) of lateral cuboid resection performed in 55 children with pathologic foot adduction. The age of the children ranged from 18 months to 12 years (average age 41 months). This residual deformity resulted from the treatment of clubfoot (both conservative and surgical). The procedure was performed in cases with severe forefoot adduction (type III according to Simondsa). Late results of treatment of 54 feet (75.9%) of the cases were assessed according to the Magone classification yielding good and very good results in 41 (75.9% feet; satisfactory in 10 (18.6%) feet. In 95% of case correction of adduction deformity was achieved. The authors stress the fact, that the degree of correction is correlated to the extent of the primary clubfoot defect. This procedure allowed correction of talo-navicular relation. The described procedure may be performed with no regard to age, although the optimal age is 3 to 8 years. PMID- 11732257 TI - [Postero-medial release in surgical treatment of congenital clubfeet: a comment to Turco's method based on personal experience]. AB - Complete correction of congenital clubfeet by conservative treatment is often impossible. Surgical treatment plays a major role in treatment of this deformity. At the Department of Pediatric Orthopedics in Lublin between 1970 and 1999 1041 children (1253 feet) were treated surgically with Turco's method, in the authors' own modification. The paper presents the technically optimal procedure, the range of tendon elongation. The way the wound is closed is particularly stressed, as well as the need to achieve muscle balance, along with a description of proper post-op care. The material was analysed as a whole, although particular attention was given to three periods: 1970-1975, 1980-1985, 1990-1995. The results were assessed according to the Turco classification, the Magone classification in accordance with the injunctions of the Scientific Committee Meeting of the Pediatric Section of the Polish Orthopedic Society held in Poznan. Good and very good results were achieved in 65-67% of the cases, while satisfactory and bad results were found in 23-30% of the cases. PMID- 11732258 TI - [Evaluation of treatment outcome in congenital clubfoot by subtalar release]. AB - This paper presents an analysis of the results of congenital clubfoot treatment by partial or complete subtalar release performed through the Cincinnati approach. Of 116 patients (142 feet) who underwent surgery in the years 1995 1996, 33 (47 feet) came in for final follow-up. The type of primary deformity was defined in only in 36 feet. There were 25 type II deformity and 1 type III deformity. Total subtalar release was performed in 39 feet. Partial subtalar release was performed in 8 feet. During the release procedure, the calcaneo cuboid joint was not opened, but stabilized with a K-wire. Clinical results were assessed according to the Magone classification. Radiological results were assessed according to the modified criteria of Scientific Committee of the XXI Meeting of the Polish Orthopedic Society held in 1976. Talo-navicular reposition was assessed according to Napiontek. Follow-up time ranged from 40 to 54 months (mean: 48 months). The clinical assessment revealed that the analyzed group as a whole gave 82.2 points (yielding a good result). Very good results were noted in 12 feet (25%), good results in 21 (45%), satisfactory results in 7 (15%) and poor results in 7 (15%). Analysis of radiological results yielded on average 2.5 points (0-7 points). Good results were noted in 45 feet (96%) and satisfactory in 2 feet. A talo-calcaneal index of less than 55 degrees was noted in 23 feet. PMID- 11732259 TI - [Treatment of congenital clubfeet by the Goldner method]. AB - The authors present an overview of Goldner's concept of clubfeet pathology and treatment. Goldner stresses the role of the talus in this foot deformity. According to this author full correction of the deformity can be achieved only by posterior, medial, plantar and lateral release. PMID- 11732260 TI - [Strategy for surgical treatment of short stature]. AB - The paper presents the role of limb lengthening in the treatment of short stature. Basing on different etiologies and psychosocial aspects of short stature indications and contraindications for treatment are presented. Functional and cosmetic indications for treatment are discussed. Pros and cons of different strategies for short stature treatment are described. Basing on available literature the range of lengthening and optimal age for starting treatment are discussed. PMID- 11732261 TI - [A new method of thumb reconstruction using big and second toe elements]. AB - The author's modification of thumb reconstruction using the big and second toe elements is presented. Two separate parts were raised on one common pedicle to form future thumb. The main element including bones, tendons and soft tissues were taken from the second toe. The flap comprising nail radix, matrix and lateral fragment of the second toe pulp was prepared and left in situ. Similar flap was raised on the hallux. Winding of the hallux flap round the second toe part allowed to form nearly normal thumb. The nail radix and matrix were harvested subperiosteally without any bone fragments of distal phalanges. Harvesting of minimum of tissues from the hallux and covering the wound by similar flap from the second toe markedly limited the big toe deformation, typical for this type of procedure. The nail plate of the second toe looked acceptable on the slightly thinner hallux. Scares were localized out of the weight bearing surfaces. Best indications for this procedure include cases when the thumb is amputated on the proximal phalanx level. Rare anatomical variation of the toe vascularity may restrict application of this method. Individual length differences between the hallux and the second toe sometimes make it difficult to put the hallux flap in the proper position on the thumb. A small hallux valgity may appear when the second toe is too short. PMID- 11732262 TI - [Changes of regional cerebral blood flow in stroke patients assessed by SPECT]. AB - The aim of our work was to evaluate changes in regional cerebral blood flow (rCBF) in patients with TIA and/or completed stroke and localisation of fotopenic foci in basal conditions and, in some cases, after stimulation pharmacological trial with acetazolamide. One hundred and fifty three patients participated in the study--105 with completed stroke and 48 with TIA. Regional CBF (rCBF) was evaluated using a single-headed rotating gamma camera (Diacam, Siemens) with high resolution collimator, after injection of 740 MBq 99mTc-HMPAO (Ceretec, Amersham). Sixteen patients with TIA underwent SPECT at baseline and after injection of acetazolamide (ACZ). Relative regional perfusion was expressed as the ratio of cerebral/cerebellar activity. In the completed stroke population, the perfusion ratios were significantly lower than those of controls and TIA at baseline (p < 0.001) and TIA after ACZ (p < 0.01). CONCLUSIONS: This study shows a significant difference in rCBF in patients with completed stroke and/or TIA and controls. In patients with TIA and rCBF similar to that observed in controls, perfusion deficits may be visible after ACZ injection. PMID- 11732263 TI - [Neurological deficits in patients with primary and secondary anticardiolipin syndrome]. AB - 27 patients (22 women, 5 men); age 17 to 56 yr. (mean age 37 yr.) were included in this study, 4 had primary antiphospholipid syndrome and 18 secondary antiphospholipid syndrome in the course of systemic connective tissue disease and in 5 cases increased levels of anticardiolipid antibodies were found which did not meet the criteria necessary for diagnosis of secondary antiphospholipid syndrome. The mean duration of the disease was 8 yrs. Among primary antiphospholipid syndrome patients two had ischaemic stroke, one migraine-like headache and seizures. 18 patients had lupus erythematosus, two mixed connective tissue disease, one rheumatoid arthritis, one Sjogren syndrome, one Behcet disease. In 55% of patients migraine-like headache, polyneuropathies, encephalophaties, stroke, seizures and vision disturbances were present. In 18.5% of patients EEG exam revealed focal lesions with tendency for generalisation. On brain stem auditory evoked potentials examination, in 11.1% of patients conductivity lesions in mesencephalon and pons were found, visual evoked potentials, in 11.1% of patients in visual tracts. In 37% of patients, neuropathy was found on EMG exam. Neurological symptoms are one of the most frequent disorders in systemic connective tissue disease associated with the presence of anicardiolipin antibodies. PMID- 11732264 TI - [Vascular risk factors in demented elderly: analysis of Alzheimer Clinic materials]. AB - In recent years evidence is increasing that vascular disease is associated with cognitive impairment and dementia. Moreover, presence of cerebrovascular disease may intensify the clinical symptoms of Alzheimer's disease (AD). The aim of the study was to determine the prevalence of vascular risk factors in age and sex matched patients with dementia. We studied 109 patients with AD and 37 patients vascular dementia (VD). DSM-III-R test for dementia, NINCDS-ADRDA guidelines for AD and NINDS-ARIEN for VD were applied. RESULTS: Mean age of dementia onset in AD and VD was 65.8 SD 7.8 and 67.4 SD 7.0 years (p > 0.05), the duration of dementia, MMS and GDS for patients with AD and VD was not significantly different. Arterial hypertension was associated in 51.3% VD and 30.3% AD (p < 0.05), hypotension in 11.1 and 23.6% respectively (p > 0.05), atrial fibrillation was similar in AD and VD, coronary artery disease was presents 64.8% AD and 46.8 VD (p > 0.05) and type 2 diabetes in 21.6% and 10.1% (p > 0.05) respectively. No significant differences in serum lipid profile were found in both groups, except two times higher incidence of normal HDL-cholesterol concentration in AD compare to VD. The relation between alcohol consumption, cigarette smoking and head trauma was similar in both types of dementia. CONCLUSION: Vascular disease and AD have to some extent a shared aetiology, and risk factors that they have in common increase the risk of both disorders independently and vascular disease is perhaps involved in the aetiology of AD. PMID- 11732266 TI - [Preoperative administration of a slow releasing somatostatin analog (SR lanreotide, BIM 23014) in patients with acromegaly in the course of GH-releasing adenoma]. AB - To evaluate the therapeutic efficacy of slow releasing analogue of somatostatin (SR-Lanreotide) in the pretreatment for GH-releasing adenomas, especially macroadenomas. During the last four years (between January 1996 and December 1999) the authors carried out 382 transsphenoidal operations for to various lesions. There were 169 acromegalic patients in this group. 82 of them received, as pretreatment, the slow releasing analogue of somatostatin (SR-Lanreotide, BIM 23014) in a dose of 30 mg every 14 days for 3 months (6 injections). There were 55 women and 27 men (range 25-68, mean age 44.8 years, SD +/- 10 years) operated on by one experienced neurosurgeon. The concentrations of serum GH--70.5 micrograms/l (range 5.3-500 micrograms/l, SD +/- 83.9 micrograms/l) and IGF-I- 1302 micrograms/l (range 610-2030 micrograms/l, SD +/- 360.7 micrograms/l) were high. Out of these 82 patients 79 had macroadenomas with suprasellar and parasellar extension. The volume of the tumours was calculated according to the formula of Di Chiro-Nelson. The mean volume of the tumour was 4146.9 mm3 (range 213.5-38595.3 mm3, SD +/- 5675.9 mm3). The response to the pretreatment suppression of the serum GH, IGF-I level and shrinkage of the tumours--were evaluated before surgery. Second MR examination was performed in 38 pretreated patients. During the Lanreotide treatment mean serum GH level decreased from 70.5 to 15.6 micrograms/l (p < 0.0001), mean serum IGF-I concentration decreased from 1302 to 787 micrograms/l and mean volume of the tumour decreased from 5662 to 2326 mm3 (p < 0.0001). During surgery, tumours were observed to be softer, had liquid consistency and were easier removed. 57 patient (69.5%) who underwent surgery had GH below 5 micrograms/l and were cured. Transsphenoidal microsurgical resection of pituitary adenomas is the primary treatment for acromegaly. Lanreotide pretreatment significantly decreased mean serum GH and IGF-I level, shrinks the tumour and make it much softer and easier to be removed. PMID- 11732265 TI - [Damage to cranial and peripheral nerves following patency restoration of the internal carotid artery]. AB - The aim of the study was an assessment of the incidence of injury to cranial and peripheral nerves as complication of patency restoration of the internal carotid artery, and analysis of the effect of peripheral nerve injury on the results of carotid patency restoration. From Oct 1987 to Sept 1999 543 procedures were carried out for restoration of patency of the internal carotid artery. After the operation hypoglossus nerve injury was found in 7 cases (1.4%), vagus injury in 9 (1.8%). Signs of exclusively recurrent laryngeal nerve damage were found in 6 cases (1.2%). Glossopharyngeus nerve was damaged in 2 cases (0.4%), transient phrenic nerve palsy as a result of conduction anaesthesia was noted in 2 cases (0.4%). Damage to the transverse cervical nerve was found in 96 cases (60%). In 2 patients (1.2%) lower position of mouth angle was due to section of the mandibular ramus of the facial nerve. In another 2 cases skin sensation disturbances were a consequence of lesion of the auricularis magnus nerve and always they coexisted with signs of transverse cervical nerve damage. IN CONCLUSION: damage to the cranial nerves during operation for carotid patency restoration are frequent but mostly they are not connected with any health risks and often they regress spontaneously. PMID- 11732267 TI - [Idiopathic autonomic neuropathy (pandysautonomia)]. AB - On the basis of current literature, clinical and neuropathologic features of idiopathic autonomic neuropathy is presented. Idiopathic autonomic neuropathy is a disease characterized by acute or subacute onset, monophasic course over a period of several years, it is often preceded by an infection. The spectrum of autonomic changes ranges from cholinergic or adrenergic dysfunction to pandysautonomia, leading to heterogeneity of its clinical features. Possible sympathetic system abnormalities found in autonomic neuropathy are: poor pupillary response to light in darkness, orthostatic hypotension leading to syncope, hypotension without compensatory tachycardia, ejaculation disturbances and vasomotor instability. Possible parasympathetic dysfunctions are: salivation and lacrimation disturbances, absent pupillary constriction to light and near gaze, gastrointestinal tract immobility and impairment of gastrointestinal function, atonic bladder with large residual volume, erectile impotence. Pandysautonomia is thought to result from an immune mediated mechanism and responds well to plasmaferesis and intravenous immunoglobin therapy leading to gradual, sometimes not full, recovery. Moreover in this article we pay attention to the clinical value of many tests like cardiovascular or pharmacological studies in the diagnosis of pandysautonomia and in differentiation of pre- and postganglionic changes. In order to diagnose idiopathic autonomic neuropathy one has to rule out a large number of diseases with autonomic dysfunction e.g.: diabetes, malignant neoplasms, acute intermittent porphyria, Shy-Drager syndrome, Riley-Day's dysautonomia, Parkinson's disease, amyloidosis and others. PMID- 11732268 TI - [Superoxide dismutase-1 (SOD-1) gene mutation-dependent mechanisms of neural degeneration in amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease involving motor neuron degeneration, occurring in sporadic and familial forms. Mutations in Cu/Zn superoxide dismutase gene (SOD-1) play a key role in the pathogenesis of the familial form in which it is present in about 20%. The mechanisms by which the mutated enzyme produces the disease are not sufficiently know. The following hypothesis are considered: oxidative damage, disorganization of neurofilaments, toxic action of intracellular aggregates, disturbed mechanisms of protein synthesis or degradation, and increased glutamic acid toxicity due to damage of EAAT 2 mRNA, transporter of this acid. It is supposed that motor neuron death is due to various mechanisms caused by SOD-1 enzyme mutations. Pathological changes suggest that biochemical processes leading to neurodegeneration in familial ALS form related or unrelated to SOD-1 mutation, and in sporadic form may be very similar. PMID- 11732269 TI - [Present condition and prospects of cerebrospinal fluid diagnostics]. AB - Despite a significant reduction in the frequency of lumbar taps done for CSF examination this method is still an important diagnostic tool. In recent years important advances have been made in the studies of the cytology and biochemistry of CSF. The introduction of new simple methods of centrifugation has increased the index of cell recovery with better possibilities of differentiation of these cells. In the biochemistry of proteins in which the analysis of immunoglobulins, albumins and other proteins, such as 14-3-3 protein, S-100, tau protein, enzymes, such as neuron-specific enolase or matrix-metalproteinase, alkaline myelin protein, beta-2-microglobulin, various cytokines, has been introduced, it is becoming a routine analysis in many CSF laboratories. The role of determination of many antibodies is increasing. Particular advances have been achieved in genetic studies, and, similarly as in other medical disciplines, explanation is expected of many not yet sufficiently clear pathological mechanisms. PMID- 11732270 TI - [Vasospasm. Part II -- Onset time, localization, etiology. Proposed management algorithm in vasospasm]. AB - Time course of vasospasm in man has a big impact on clinical pisture, it also has an influence on the decision of surgery. The presence of acute stage of vasospasm is still under discussion. Many authors deny its existence. It is well known that positive decision about early operating has a crucial role in case of vasospasm Another problem is the aetiology of vasospasm. Inspite of many clinical and experimental investigations the aetiology of vasospasm remains unclear. The main problem is still the adequate therapy. This paper contains the updated knowledge about vasospasm, with the main accent on aetiology diagnosis and therapy. PMID- 11732271 TI - [Spontaneous intracranial hypotension: case report]. AB - Intracranial hypotension (IH) is a rare cause of acute headache. There are secondary IH (more frequently, after lumbar puncture, head and back trauma, brain and spine surgery) and essential (spontaneous intracranial hypotension SIH). SIH was first described by Schaltenbrand in 1938. This syndrome is characterized mainly by acute postural headache. The authors report a case of 45-year-old women with SIH. Clinical diagnosis was confirmed by lumbar puncture and characteristic MR. The symptoms completely regressed within 2 weeks. The treatment was based on bed rest, hypotonic solutions, steroids, caffeine. Pathophysiology, diagnosis and treatment of SIH are discussed. PMID- 11732272 TI - [Cranial epidural abscesses: a case report]. AB - OBJECTIVE AND IMPORTANCE: Cranial epidural abscess mostly follows sinusitis or mastoiditis, also open cranial injury or cranial surgery with iatrogenic infection. In our case the abscess possibly evolved from epidural haematoma and the process invaded frontal sinus and subgaleal space in the frontal region. CLINICAL PRESENTATION: A 22-year-old male after motorcycle accident 4 years ago, with signs of sinusitis frontalis and seizures, with CT confirmed epidural abscess. INTERVENTION: Craniotomy was performed and the abscess totally removed. Homogeneous bone was replaced at the operation. Good result, after 2 months without pathologic signs and correct postoperative tomography. CONCLUSION: We conclude on the basis of disease history and clinical evidence, that the most probable was transformation of the abscess from epidural haematoma end in long term period perforated through the frontal bone to the frontal sinus, and to the subgaleal space with signs of sinusitis at last. PMID- 11732273 TI - [Obituary: P. E. Becker (1918-2000): life and work]. PMID- 11732274 TI - [Report on the Neurosurgical Microvascular workshop in Ceske Budejovice, Czech Republic, October 26-27, 2000]. PMID- 11732275 TI - [The effect of selegiline and vitamin E in the treatment of ALS: an open randomized clinical trials]. AB - A role for oxidative stress in the etiology or progression of amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases has been recently proposed. We conducted the 18-month, randomized treatment trial with oral vitamin E (600 IU daily) and selegiline (10 mg daily) in 67 patients with sporadic ALS. Thirty five patients were randomly assigned to receive antioxidative therapy (vitamin E plus selegiline) and the remaining 32 patients were the ALS controls who received symptomatic treatment. The primary end point was survival and functional status. At the end of 18-month study, 13 patients in the treatment group and 14 in the control group died or were tracheostomized. A decline in functional disability was also similar in both groups. Long-term antioxidative treatment did not benefit patients with ALS. PMID- 11732276 TI - [Kennedy's disease: expansion of the CAG trinucleotide]. AB - Kennedy's disease is a rare X-linked spinal and bulbar muscular atrophy (SBMA). A degenerative process of the motor neurons is associated with an increase in the number of CAG repeats encoding a polyglutamine stretch within the androgen receptor. Despite a distinctive clinical phenotype, SBMA can be misdiagnosed, usually due to the lack of clear family history. Accurate diagnosis is important for genetic counseling and because alternative diagnosis of amyotrophic lateral sclerosis usually means much worse prognosis. We report 2 unrelated patients with Kennedy's disease in whom the clinical diagnosis was confirmed by showing the CAG repeat expansion. PMID- 11732277 TI - [Electrophysiological investigation s in diagnosis and evaluation of ALS progress]. AB - In diagnostics of ALS the electrophysiologic investigation should be able to evaluate two fundamental processes: the primary process of the loss of some motoneuros--denervation, and secondary process of reinnervation. The most important diagnostic methods include the quantitative electromyography (EMG) evaluating several parameters of the motor unit potential (MUP) and of the maximal effort pattern. The earliest features are the signs of spontaneous activity (denervation) and the elevation of MUP amplitude and area (reinnervation). Finding of spontaneous activity in the tongue muscles as well as in the paraspinal muscles is of great diagnostic value. SFEMG may early detect increased density of muscle fibres (FD) and jitter elongation as signs of recent reinnervation. Electroneurography is critical in differentiation against the multifocal motor neuropathy with conduction block and other polyneuropathies. The aim of the electrophysiological investigations is also the evaluation of the intensity of lesion in the muscle investigated and, indirectly, the evaluation of the progress of the disease in serial studies. It might play a great role in prognosing and in monitoring of therapeutical trials. The abnormalities in the maximal effort pattern presenting as decrease of recording density and amplitude as well as of bioelectric activity indicate in the quantitative EMG a high degree of lesion. Those signs are accompanied by altered MUP parameters: decrease of MUP amplitude and area as compared with the former stages of reinnervation. Those signs express decompensation and denervation predominating over reinnervation. In SFEMG they present as FD decrease and in Macro-EMG as decrease of amplitude as compared to the reinnervation period. Serial investigations of amplitude of the potential resulting from supermaximal stimulation (CMAP), quantitative evaluation of the maximal voluntary isometric contraction (MVIC) and the assessment of the number of motor units (MUNE) are further valuable methods in monitoring of ALS progress. PMID- 11732278 TI - The pathogenesis of ALS: implications for treatment strategies. AB - Besides the free radical hypothesis raised by the identification of Superoxide Dismutase I mutations in a subset of familiar Amyotrophic Lateral Sclerosis (ALS) patients, three etiopathogenic hypotheses for sporadic ALS, namely autoimmune, neurofilament, and glutamate toxicity, have attracted interest in the last few years. The role of autoimmunity in ALS has been seriously questioned. The excitotoxic hypothesis for ALS spurred two clinical trials with riluzole. The results of both studies showed a modest benefit in prolonging survival that was statistically significant. Riluzole was the first drug made available for ALS patients. It began a new era in both basic and clinical research. Various human recombinant neurotrophic molecules (CNTF, BDNF, IGF-I) were administered to ALS patients. IGF-I slowed the progression of functional impairment in patients with ALS with no adverse effects. The recent demonstration of the specific viral echovirus 7 RNA sequences in the spinal cord of ALS patients refocused research on the viral hypothesis of the disease and antiviral drugs are ready to be used in clinical settings. New treatment strategies are today under study: intrathecal infusion with BDNF, intrathecal capsules for neurotrophic factor secretion or in vivo gene therapy using viral vectors. New research findings are, more than for other diseases, immediately transferred to clinical ground for the desperate need of a curative treatment of the patients affected by ALS. PMID- 11732279 TI - Pharmacological treatment of ALS. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease resulting from chronic and selective loss of motor neurons in the brain and spinal cord. In 1993, the etiology of ALS has been clarified for a small sub-group of patients with an autosomal-dominant form of this disease. About 10 percent of familial ALS patients have been associated with more than 50 mutations of the gene of the Cu/Zn superoxide dismutase (SOD1). Mutations in the SOD1 gene account for 1 percent of all ALS patients and have therefore limited epidemological and clinical relevance; however, they are of fundamental importance for the understanding of the ALS pathogenesis, and the development of neuroprotective strategies. In two double-blind and placebo-controlled studies the membrane stabilisator riluzole has been shown to be the first neuroprotective compound with a significant effect on survival of ALS patients. The neuroprotective approach reduced therapeutic nihilism in ALS and is a first step in the treatment of this devastating disease. PMID- 11732280 TI - [Symptomatic treatment and palliative care of ALS]. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, affecting upper and lower motor neurons, which eventually progresses to respiratory deterioration and death in most of the patients. Only one drug, riluzole, has been approved for the treatment of ALS. The drug has a benefit, prolonging life by 3-6 months, but the disease progresses inexorably, with no better quality of life. The fundamental role of medicine is sometimes to cure, but always to bring comfort. In current situation, ALS patients need adequate palliative care more than anything else. Prognosis and treatment options should be discussed with the patient and the relatives, but full information about the prognosis may deprive the patient of hope. However, disclosure of the prognosis is necessary to obtain informed consent for management decisions such as tracheostomy and artificial ventilation. Nasal positive-pressure ventilation (BiPAP) is an alternative to tracheostomy, at least for some patients without advanced bulbar impairment. Nutritional status in patients who cannot swallow can be efficiently improved by a percutaneous endoscopic gastrostomy. (PEG). PMID- 11732281 TI - [Proton MR spectroscopy studies of the brain in ALS patients]. AB - Nuclear magnetic resonance spectroscopy (MRS) has an ability to measure brain metabolites noninvasively in vivo. The content of N-acetyl-aspartate (NAA) is used as a biochemical marker of neuronal integrity and viability. In amyotrophic lateral sclerosis (ALS) patients the degeneration and neuronal loss of motor cortex was reported. The presence of these changes can lead to the decrease of NAA. The aim of this study was to evaluate the neurochemical status of motor cortex (by using 1H-MRS) in 11 patients with clinically definite or probable forms of ALS (according to the El Escorial criteria). These data were compared with MRS results from 4 healthy controls. The mean NAA/(Cr + PCr) values were decreased by 19% (p < 0.05) when compared with controls. In 4 ALS patients the NAA/(Cr + PCr) ratio was decreased by 31% (p < 0.05) and these patients had rapidly progressing disease. In other 7 ALS patients, we found that NAA/(Cr + PCr) was decreased by 10% (p < 0.05) and they showed less advanced neurological symptoms. 1H-MRS of the motor cortex can be a new diagnostic tool in ALS and it might help to monitor the progress of the disease. PMID- 11732282 TI - [Corticospinal tract assessment in ALS: transcranial magnetic stimulation]. AB - Results obtained during transcranial magnetic stimulation (TMS) in 79 patients with ALS were compared to those obtained in 10 healthy volunteers. M and F responses evoked by peripheral stimulation of the ulnar and peroneal nerves and then motor evoked potentials (MEP) induced by TMS were recorded in the abductor digiti quinti (AV) and tibial anterior (TA) muscles bilaterally. Central motor conduction time (CMCT) was calculated using MEP and F response latencies. The MEP/M x 100% amplitude ratios were calculated. Mean CMCT was significantly (p < 0.0001) prolonged in patients with ALS (9.6 +/- 4.1 ms for AV and 20.4 +/- 9.6 ms for TA) and MEP/M x 100% amplitude ratios were higher (45 +/- 75% for AV-p < 0.001 and 27 +/- 32.7% for TA p = 0.2). In a sub-group of patients with predominant upper motor neuron (UMN) involvement, the CMCT was significantly prolonged (14.2 +/- 5.8 ms for AV and 28.5 +/- 9.1 ms for TA), but MEP/M x 100% amplitude ratio was 24.1 +/- 37.6% for AV and 11.4 +/- 12.9% TA. These results confirm usefulness of combined analysis of CMCT and MEP/M x 100% amplitude ratio as the method of assessment of pyramidal tract function in ALS. PMID- 11732283 TI - [Neurotoxic activity of serum and cerebrospinal fluid of amyotrophic lateral sclerosis patients against some enzymes of glutamate metabolism]. AB - One of the hypotheses in amyotrophic lateral sclerosis (ALS) indicates on excitatory amino acids as the cause of neuronal death. Changes in their concentration in the tissues and body fluids may be the consequence of a defect in their transport, as well as abnormal activities of glutamate metabolizing enzymes. Abnormal synthesis/degradation of these enzymes and/or influence of activators/inhibitors should be taken into account. The activity of enzymes of glutamate metabolism of rat spinal cord in vitro in the presence of serum and cerebrospinal fluid (CSF) of 20 patients with ALS and 20 healthy controls was tested. In the presence of serum of the ALS patients glutaminase was significantly stimulated, instead of being inhibited; the inhibition of GABA aminotransferase, glutamate decaboxylase and aspartate aminotransferase was less evident than in the controls, glutamate dehydrogenase lost its activity more than in control conditions, the inhibition of glutamine synthetase was comparable to that when normal serum was applied. The activity of the enzymes in the presence of CSF of ALS patients was generally similar to that of normal CSF, except of glutaminase which was stimulated and GABA aminotransferase, which was inhibited stronger than in the presence of normal CSF. This study indicates, that changes in glutamate concentration in tissues and body fluids in ALS may be caused, at least partly, by abnormalities in the activity of glutamate metabolism enzymes, which are in turn induced by neurotoxic agents present in body fluids of ALS patients. PMID- 11732285 TI - [Etiology of pleural effusion based on material from the Hospital of Lung Diseases and Tuberculosis]. AB - The aim of this study was to show the influence of the changes in incidence of tuberculosis and lung cancer on the etiology of pleural effusion among the patients of the Hospital of Lung Diseases and Tuberculosis. To this end 349 patients with pleural effusion were separated from the total number of 9786 patients treated in years 1990-1999. The most numerous group--180 patients (51.58%)--consisted of the cases of malignant neoplasms, 109 (31.25%)- tuberculosis and 60 (17.17%)--remaining diseases altogether. At the time, the statistically significant increase of the whole number of patients with pleural effusion as well as the increase of patients with pleural effusion caused by malignant tumors were recorded. It was shown that the increasing incidence of lung cancer in Silesia at that time caused the change of the profile of the Hospital of Lung Diseases and Tuberculosis. PMID- 11732284 TI - [The role of mitochondrial respiratory chain in the pathogenesis of ALS]. AB - Mitochondrial dysfunction and abnormal electron chain transport (ECT) may be involved in the pathogenesis of ALS. The aim of this study was to investigate the effect of cerebrospinal fluid (CSF) from ALS patients on the activity of ECT enzymes in mitochondrial cerebral crude preparations in the rats. We found that CSF inhibited the activity of complex I-III in 20%, complex II-III in 12% and complex IV in 33% of the ALS patients. CSF from the controls did not affect the activity of complex I-III and II-III. The effect of the CSF ultrafiltrates with cut off below 5000 daltons on the activity of ECT enzymes was also investigated. The CSF ultrafiltrates inhibited the activity of complex I-III, complex II-III and complex IV in 38%, 44% and 53% of the ALS patients, and in 80%, 53% and 43% of the controls, respectively. The results of this study and our previously reported experiments on the sera of ALS patients may indicate that neurotoxic effects of body fluids from ALS patients could be mediated by inhibition of the respiratory chain enzymes. This confirms an important role of mitochondrial dysfunction in the pathogenesis of ALS. PMID- 11732286 TI - [Telepathology in diagnostic consultations for controversial neoplastic growths in the lung--personal experience]. AB - The authors assessed the diagnostic accuracy of the static telepathology (sTP) for practical consultations in the controversial pulmonary oncology cases. The short characteristics of the diagnostic cases is included. We reported the results of 6 difficult clinico-pathological cases submitted to Dept Quantitative Pathology [DQP] for consultations (5F and 1M, age: 26-68 yrs). Digital images of histological or cytological samples were captured at DQP and transmitted to telepathologist (TPat) in Thoraxklinik, Heidelberg or AFIP, Washington DC. Simultaneously, the same slides (or with a paraffin block) were mailed to TPats for re-evaluation with the conventional microscope (dgn-zwPat). The controversial cases presented 2 types of diagnostic problems: 1/rare entities, 2/common difficulties in the routine work of pathologist, but with indefinitely proved diagnosis. The first group encounters TPat diagnoses as follows: * case A: pulmonary plasma cell granuloma with lymphoidal interstitial infiltrates [LIP]- preleukemia; * case B: microfoci of early metastases of benign uterine leiomyoma; * case D: small cell carcinoma spreading along pleura. The second group included: * case C: invasive epidermoidal carcinoma in bronchus; *case E: probably metastatic adenocarcinoma of colon; *case F: synchronous or metastasising 2 lung tumours sharing NE morphology or NE immunohistochemical features. There was very high concordance between referring pathologist (ref-Pat) diagnoses and TPat diagnoses as well as a consensus in zw-TPat diagnoses. In 2 cases the conceptual problem was solved by TPat. Telepathology offered a support or improved the quality of the final diagnosis. The implementation of sTP remarkably reduced the time of consultations and allowed to present the diagnostic problems to the international group of experts. PMID- 11732287 TI - [Montelukast in treatment mild chronic asthma]. AB - The efficacy of montelukast, a leukotriene receptor antagonist, in treatment of mild asthma was evaluated. METHODS: Thirty children aged 6 to 14 years with mild persistent asthma (asthma history more than 12 months and > or = 15% FEV1 improvement after inhaled beta 2-agonist in the past, good control of asthma with inhaled cromolyn or budesonide in the last three months) were enrolled. The study included three periods (2 week's each): washout, placebo, and montelukast. Asthma symptoms score (range 0-5) and PEF were estimated twice daily by children. Spirometric parameters FEV1 and MEF50 were measured during three consecutive visits: on the day of study inclusion, on the last day of the placebo and montelukast period. RESULTS: The mean value of asthma symptoms score was significantly lower during the montelukast period in comparison with placebo (p = 0.038). The mean PEF values were significantly higher during the montelukast vs. placebo period (p = 0.0091). Moreover, in the montelukast period, the mean PEF values in the second week were significantly higher than those in the first week (p = 0.003). The mean FEV1 predictive value in the last day of the montelukast period was higher, though not significantly, than on the day of study inclusion and on the last day of the placebo period. A similar change in mean MEF50 values was observed. CONCLUSION: In children aged 6-14 years with mild persistent asthma, montelukast treatment significantly diminishes asthma symptoms and increases mean PEF values comparing to placebo. PMID- 11732288 TI - [Molecular studies in a population of children with bronchial asthma. I. Polymorphism in the promotor region of gene CD14]. AB - Allergic asthma is associated with the recruitment of the inflammatory cells into the bronchial mucosa. Surface expression of CD14, a marker of activation and differentiation of macrophages/monocytes, was suggested to protect against Th-2 response. OBJECTIVE: To investigate clinical effects of genetic polymorphism of CD14 promoter in asthmatic children. METHODS: In ISAAC survey, 50 children with asthma were identified (wheezing in the last year, serum IgE level > 150 kIU/l, positive bronchial challenge test with aerolized hypertonic saline) and 73 children without the above signs. Age range of surveyed children was 13-14 years. Genotypic pattern of CD14 promoter -159 C to T transition was assessed by RFLP method. RESULTS: There was no difference in the allelic (0.36 vs. 0.38 for -159T) or genotype frequencies (0.12 vs. 0.15 for -159TT) of CD14 polymorphism between allergic asthmatics and controls. Moreover, there was no relationship between CD14 genotype and serum IgE level or bronchial hyper-responsiveness. CONCLUSION: Our results do not confirm the association of CD14 polymorphism (promoter -159 C to T transition) with asthma in Polish children. PMID- 11732289 TI - [Pulmonary Langerhans cell histiocytosis--clinicopathologic analysis of 7 cases]. AB - Langerhans cell histiocytosis with only pulmonary involvement is a rare, probably reactive, proliferating disease of young and smoking persons. During last 7 years 7 cases of Langerhans cell histiocytosis were diagnosed in our Department. There were 6 men and one woman. Mean age was about 32 years. All of them were smokers. Intensity and duration of smoking varied widely from 5 to 40 cigarettes/day and 5 to 30 years respectively. The most frequent symptoms were dry cough, dyspnea. Two patients had no complaints. Pneumothorax preceded diagnosis in 4 patients. Duration of symptoms varied from 3 months to 10 years--in a patient who was previously diagnosed as allergic alveolitis. All patients had typical radiological and HRCT appearance of disseminated nodular lesions with small cysts formation with sparing of costo-phrenic angles. Three of patients had very advanced disease with cystic lung destruction. Restriction was noticed in those three patients. Diminished values of compliance and diffusing capacity for CO was observed in four patients. Bronchoscopy examination of bronchial tree revealed only slight inflammation in all patients. BAL was assessed for presence of CD1a cells in 6 cases, but in no one patient elevated number of those cells was noticed (2-4.5%). In all patients open lung biopsy was performed, and typical histologic appearance was observed. In six patients also CD1a positive cells were revealed in lung specimens. PMID- 11732290 TI - [Pulmonary alveolar microlithiasis in a patient with urolithiasis and cholelithiasis]. AB - 49 years old woman was admitted to Pulmonary Ward because of respiratory and cardiac failure. Six years earlier she was surgically treated because of urolithiasis and at this time disseminated lesions in both lungs were revealed. Biopsy made during transbronchial procedure showed microlithiasis. She had'nt any respiratory symptoms during 6 years. Now she had PaO2 = 36 mmHg and massive infiltrations in whole lungs. Palliative therapy with diuresis, cardiac drugs and oxygen therapy diminished symptoms of respiratory and cardiac failure. PMID- 11732291 TI - [Primary lung hemangiopericytoma--a rare neoplasm with a long course of recurrence over many years]. AB - Hemangiopericytoma (HPC) is a rare neoplasm arising from pericytes that occur mostly around smaller vessels. Up to now only about 100 cases have been reported to arise primarily in the lung. The behavior of pulmonary hemangiopericytomas is difficult to predict and all tumors should be considered potentially malignant, even though the criteria for malignancy are not well developed. The diagnosis of HPC is known to confound even experienced pathologist. Pericytes lack readily identifiable morphologic features, therefore hemangiopericytomas are often confused with other soft tissue tumors that may have hemangiopericytoma--like pattern. We report a rare case of primary HPC of the lung with an asymptomatic, long course of the disease. The diagnosis of hamartoma was established after the first operation. Subsequently, seven years later, a chest radiograph revealed new lesions. Histological examination, including immunohistochemistry lead to the final diagnosis of hemangiopericytoma. The small number of cases of primary pulmonary hemangiopericytoma makes it difficult to define the correct histopathological diagnosis especially without modern methods. PMID- 11732292 TI - [A case of tracheal adenoid cystic carcinoma in a young woman]. AB - A case of an adenoid cystic carcinoma of trachea is presented. A 30-year-old non smoking woman with strong inspiratory dyspnea at rest was admitted to the Dept. of Pulmonary Diseases. At auscultation a respiratory murmur was more silent at right lung and stridor over trachea was heard. CT scan revealed tumor at the bifurcation of the trachea. Bronchoscopy was made and biopsy established the diagnosis: adenoid cystic carcinoma. The tumor was partially removed with rigid bronchoscope and radiotherapy was started. Clinical improvement occurred; in control CT scan tumor vanished. The trachea cancers are rare. Symptoms often mimic asthma or chronic bronchitis. Thus in every patient with chronic cough and dyspnea bronchoscopy should be made. A treatment of choice is primary resection and postoperative radiotherapy. PMID- 11732293 TI - [Surgical treatment of limited small cell lung cancer]. PMID- 11732294 TI - [Angiogenesis in non-small cell lung cancer--clinical implications]. PMID- 11732295 TI - Tuberculous ureteric stricture. AB - The management of a young woman with a ureteric stricture is a challenge to the urologist. Genitourinary tuberculosis can cause ureteric strictures, with minimal symptoms. Untreated, it can lead to impairment of renal function. Diagnosis of genitourinary tuberculosis may be delayed until histopathological assessment. Drug treatment and reconstructive surgery produce good results. PMID- 11732296 TI - A man with thyrotoxicosis, lymphoma and thymic hyperplasia. AB - We report a case of non-Hodgkin's lymphoma of Burkitt's type with thyrotoxicosis and thymic hyperplasia in a 27-year old man. As far as we are aware this is the first reported case of Burkitt's lymphoma, thymic hyperplasia and thyrotoxicosis occurring in the same patient. PMID- 11732297 TI - Congenital epulis in a neonate. PMID- 11732298 TI - Dengue fever associated with a haematoma of the rectus abdominis muscle. AB - We describe a patient with dengue fever with a haematoma of the rectus abdominis muscle, due to a tear in the inferior epigastric artery following episodes of retching. From the literature reviewed, we noted that there are no previous reports of dengue fever associated with muscle haematomas. PMID- 11732299 TI - Blood lead levels of children living near Borella junction. PMID- 11732300 TI - Advances in mother-to-child transmission of HIV infection. PMID- 11732301 TI - Antibodies to hepatitis C virus in patients who have had multiple transfusions in Sri Lanka. AB - OBJECTIVE: To determine the prevalence of antibodies to hepatitis C virus (HCV) in patients who have had multiple transfusions in Sri Lanka. SETTING: University Medical Unit at the National Hospital, Colombo, the Cancer Institute, Maharagama, and the Lady Ridgeway Children's Hospital, Colombo. PATIENTS: One to 5 ml of blood for serology was collected from 200 multiply transfused patients (those who have received five or more blood transfusions). METHOD: The sera were tested for HCV specific antibodies using a third generation anti-HCV enzyme immuno-assay (EIA) kit. All sera giving positive or intermediate EIA results were re-tested by a commercial HCV Western blot confirmatory test. RESULTS: Of the 200 patients, 10 (5%) were repeatedly positive and confirmed by the Western blot. 33% (7/21) of haemophiliacs and 10% (3/31) of thalassaemics were positive for antibodies to HCV. Antibodies were not detected in other groups of multiply transfused patients (haemolytic disease, aplastic anemias, chronic renal failure, haematological and other malignancies). Of the 200 patients, those who have had more than 80 blood transfusions had a significantly higher prevalence of antibodies to HCV. The frequency of HCV infection was also higher among those who had received factor concentrates. CONCLUSION: 33% of haemophiliacs and 10% of thalassaemics who have received multiple transfusions were infected with HCV. These findings warrant a larger study among blood donors, and justify screening and decontamination of blood and blood products given to haemophiliacs and thalassaemics in Sri Lanka. PMID- 11732302 TI - Breast examination of older women in a teaching hospital. AB - INTRODUCTION: Breast examination during routine physical examination may help detect breast cancer and effect early treatment. OBJECTIVE: To determine whether doctors routinely perform breast examination in older women, and to assess attitudes of patients and doctors to this examination. METHODS: A questionnaire based survey of 150 women over 65 years attending a teaching hospital, and 51 doctors working in this hospital. RESULTS: Very few women had a breast examination performed by a doctor. All thought breast examination was important, and would give consent for this examination. Although the great majority of doctors thought breast examination should be done routinely only very few do so. CONCLUSION: Older women have a positive attitude towards breast examination, but this is not reflected by the practice of doctors. There is a need for change in attitudes and training among doctors so that breast examination would be performed routinely. PMID- 11732303 TI - Prospective study of congenital heart disease in children. AB - OBJECTIVE: Descriptive analysis of patients with congenital heart disease. DESIGN: Prospective study. SETTING: Professorial Paediatric Unit, Lady Ridgeway Hospital, Colombo 8. METHOD: All patients with congenital heart disease (CHD) admitted to the Professorial Paediatric Unit at Lady Ridgeway Hospital from August 1998 to August 1999 were included in this study. Data were collected using a pretested questionnaire which was filled by one of the authors. Patients were followed up to date. RESULTS: 102 patients, 53 (52.0%) males with congenital heart disease were studied. All patients were evaluated with chest xray, ECG and 2 D echocardiography, except for two. Based on clinical and echocardiographic findings 69 (67.6%) were found to have acyanotic and 33 (32.4%) cyanotic heart disease; 22 (21.5%) had a ventricular septal defect. Tetralogy of Fallot was diagnosed in 10 (9.8%) patients and was the commonest cyanotic CHD. 52 (51.0%) cases were detected between the ages of 2 weeks to 12 months, and 39 (38.2%) were detected routinely at the postnatal examination of the newborn. Only 12 (11.7%) patients underwent corrective surgery, 10 in Sri Lanka, and 2 in India. 19 children died during the study period, a case fatality of 18.6%. CONCLUSIONS: Previous studies have shown that congenital heart disease accounted for between 16% to 30% of deaths in the Professorial Paediatric Unit at LRH. This is the second commonest cause of deaths in the unit. This study has revealed a high case fatality rate among the patients awaiting corrective surgery. Unless surgical care for patients with congenital heart disease in the public sector are improved, many such children will continue to die early due to lack of facilities. PMID- 11732304 TI - Fish bone migration. PMID- 11732305 TI - Central neurogenic hyperventilation in an awake patient. PMID- 11732306 TI - Sex chromatin and nucleolar analyses in Rumex acetosa L. AB - Rumex acetosa (sorrel) is a dioecious plant with a XX/XY1Y2 sex chromosome system. Both the Y chromosomes are nearly entirely heterochromatic and it has been hypothesised that they can persist as chromocenters in male interphase nuclei. Using specific antibodies against 5-methylcytosine and histone H4 acetylated at terminal lysine 5, global levels of DNA methylation and histone acetylation were studied on the sex chromosomes and autosomes of both sexes. The heterochromatic Y chromosomes did not display a higher methylation level compared to the autosomes. The only prominent hypermethylation signals were found at two nucleolar organising regions located on the autosome pair V, as confirmed by in situ hybridisation with 25S rDNA probe and staining. Immuno-analysis of DNA methylation on female and male interphase nuclei neither revealed any sex specific differences. Two active (silver-positive) nucleoli and two likely inactive nucleolar organising regions (displaying prominent methylation signals) were found in both sexes. In a fraction of nuclei isolated from leaf cells, two peripheral bodies strongly positive for 4',6-diamidino-2-phenylindole were observed only in males, never in females. These heterochromatin regions were depleted in histone H4 acetylation at terminal lysine 5 and corresponded, according to in situ hybridisation with a Y-chromosome-specific repetitive probe, to the two Y chromosomes. We conclude that the peripheral condensed bodies observed exclusively in male nuclei represent the constitutive heterochromatin of the Y chromosomes which is characterised by a substantial histone H4 underacetylation. PMID- 11732307 TI - Microtubule organization in root cells of Medicago truncatula during development of an arbuscular mycorrhizal symbiosis with Glomus versiforme. AB - The colonization of plants by arbuscular mycorrhizal fungi has been shown to induce changes in cytoplasmic organization and morphology of root cells. Because of their role in a variety of cellular functions in plants, it is likely that microtubules are involved either in the signaling events leading to the establishment of the symbiosis or in changes in host cell morphology and cytoplasmic architecture. Recent studies of the arbuscular mycorrhizal symbiosis have shown that root cortical cells reorganize their microtubules upon colonization. These studies, however, have focused primarily on the cells containing hyphal coils or arbuscules and did not include descriptions of microtubule changes in adjacent cells. To probe further into the potential role of the microtubule cytoskeleton in the establishment of arbuscular mycorrhizal symbiosis, we examined the three-dimensional arrangement of microtubules in roots of the model legume Medicago truncatula colonized by the arbuscular mycorrhizal fungus Glomus versiforme by indirect immunofluorescence and confocal microscopy. Our data show extensive remodeling of the microtubule cytoskeleton from the early stages of arbuscule development until arbuscule collapse and senescence. While confirming some of the microtubule patterns shown in other mycorrhizal systems, our results also reveal that cortical cells adjacent to those containing arbuscules or adjacent to intercellular hyphae reorganize their microtubules. This indicates that the cortical cells initiate the modification of their cytoskeleton prior to entry of the fungus and is consistent with signal exchange between the symbionts prior to fungal penetration of the cells. PMID- 11732308 TI - Phallacidin stains the kinetochore region in the mitotic spindle of the green algae Oedogonium spp. AB - We found previously that in living cells of Oedogonium cardiacum and O. donnellii, mitosis is blocked by the drug cytochalasin D (CD). We now report on the staining observed in these spindles with fluorescently actin-labeling reagents, particularly Bodipy FL phallacidin. Normal mitotic cells exhibited spots of staining associated with chromosomes; frequently the spots appeared in pairs during prometaphase-metaphase. During later anaphase and telophase, the staining was confined to the region between chromosomes and poles. The texture of the staining appeared to be somewhat dispersed by CD treatment but it was still present, particularly after shorter (< 2 h) exposure. Electron microscopy of CD treated cells revealed numerous spindle microtubules (MTs); many kinetochores had MTs associated with them, often laterally and some even terminating in the kinetochore as normal, but the usual bundle of kinetochore MTs was never present. As treatment with CD became prolonged, the kinetochores became shrunken and sunk into the chromosomes. These results support the possibility that actin is present in the kinetochore of Oedogonium spp. The previous observations on living cells suggest that it is a functional component of the kinetochore-MT complex involved in the correct attachment of chromosomes to the spindle. PMID- 11732309 TI - Characterization of intracellular bacteria in the freshwater dinoflagellate Peridinium cinctum. AB - Intracellular bacteria belonging to two phylogenetically different groups of eubacteria were found in cultures of the freshwater dinoflagellate Peridinium cinctum (O.F. Muller) Ehrenberg isolated from the eutrophic lake Plusssee (Federal Republic of Germany). The phylogenetic relationships of the bacteria were studied with fluorochrome-conjugated oligonucleotides specific for archaebacteria, eubacteria, alpha-, beta- and gamma-proteobacteria, complementary to 16S rRNA and 23S rRNA sequences, respectively. The bacteria are members of the eubacterial alpha- and gamma-subgroups of proteobacteria. PMID- 11732310 TI - Population differences in the expression of nucleolus organizer regions in the grasshopper Eyprepocnemis plorans. AB - Fluorescence in situ hybridization revealed the presence of ribosomal RNA genes in paracentromeric regions of all A chromosomes and in the distal half of B chromosomes in embryonic cells from Moroccan specimens of the grasshopper Eyprepocnemis plorans. The expression of these genes was monitored by the presence of nucleoli attached to each chromosome bivalent in diplotene cells from males collected from two different Moroccan populations and was compared to previous data of Spanish populations. Whereas only the nucleolus organizer regions (NORs) on S9-S11 and X chromosomes were active in the Spanish specimens, Moroccan individuals showed NOR activity in all chromosomes. The rRNA genes on the B chromosome were inactive in both populations. The S9 and S10 NORs were less active in Moroccan specimens than in Spanish specimen, which might be partly explained by the negative inter-dependence for expression of the S10 NOR with respect to those on L2 and X chromosomes. On the other hand, the X NOR was more active in Moroccan specimens than in Spanish specimens, and this might be partly due to the positive effect that the presence of B chromosomes has on the expression of this NOR. The implications of these observations on current models of NOR activity regulation are discussed. PMID- 11732311 TI - Suppression of spermatogenesis for cell transplantation in adult mice. AB - Spermatogenesis occurs within the testis of adult males by a complex and very well organized process. Breakthroughs in techniques such as cryopreservation and culture of spermatogenic cells and the maturation of these cells in exogenous testes after transplantation renewed the interest in this process. Transplantation of spermatogenic cells from a donor to a recipient animal needs a preparatory step that consists in the elimination of the endogenous population of spermatogenic cells. The most common method used to empty the seminiferous tubules is the treatment with busulfan (1,4-butanediol dimethanesulfonate). Busulfan partially eliminates stem cells because of its alkylating nature, but a residual component of stem cells survives the treatment and competes in the regeneration of the testis with transplanted cells. Estradiol has also been used as an agent that causes a delay in the process of spermatogenesis by altering its hormonal stimulation, although it does not affect the spermatogonia population. Therefore, we have tested different treatments with busulfan, estradiol benzoate, and also an agonist of the chorionic gonadotrophin-releasing hormone, leuprolide acetate, for the inhibition of endogenous spermatogenesis. We have found that a combination of estradiol, busulfan, and leuprolide can destroy the population of endogenous spermatogenic cells without altering Sertoli cells and maintains the optimal environment needed to allow the development of transplanted cells. PMID- 11732312 TI - Expression of different abscisic acid-responsive genes during somatic embryogenesis in sugarcane (Saccharum officinarum). AB - We have examined the expression of four genes during somatic embryogenesis in two cultivars of sugarcane, Saccharum officinarum, one drought resistant (JA-605) and the other one sensitive (C-8751), as well as in embryogenic and nonembryogenic tissues treated with abscisic acid (ABA) or after drought stress. Three of the cDNAs probed, from plasmids pMA1049, pMA2005 and CM1, were lea genes and the other one, from a barley hemoglobin gene, was induced in other species under stress conditions. Only transcripts homologous to the pMA2005 and CM1 clones were differentially accumulated during somatic embryogenesis; moreover, the CM1 clone only appeared in somatic embryos of the non-resistant variety. All the lea cDNAs were dramatically increased in the embryogenic tissues treated with ABA, but we observed differences among the accumulation of these mRNAs in the nonembryogenic tissues treated with ABA, and only the pMA2005 transcript appeared. The mRNA homologous to the barley hemoglobin gene appeared in the nonembryogenic tissue and the expression increased after ABA treatment. The desiccation treatments had different effects on the varieties, and the pMA2005 transcripts only appeared in the sensitive variety. PMID- 11732313 TI - Loading process of sugars into cabbage petiole and asparagus shoot apex cells by incubation with hypertonic sugar solutions. AB - The freezing tolerance of cabbage petioles and asparagus shoot apexes was increased by preincubation with 0.8 M sugar solutions. In cabbage petioles with an initial freezing tolerance of -3 degrees C (temperature for 50% cell survival), as determined by both electrolyte leakage and fluorescein diacetate vital staining, the freezing tolerance was increased to -13 degrees C by incubation with sorbitol solutions for 3 h. In meristematic cells of asparagus shoot apexes with an initial freezing tolerance of -7.5 degrees C, as determined by fluorescein diacetate vital staining, the freezing tolerance was increased to 30 degrees C by incubation with 0.8 M sugar solutions for 3 h, although other cells in the shoot apexes were killed by higher freezing temperatures. During incubation of both cabbage petioles and asparagus shoot apexes with sugar solutions, sugars were intracellularly taken up by osmotically induced fluid phase endocytotic vesicles, as indicated by comovement of Lucifer Yellows carbohydrazide (LYCH) observed with a confocal laser scanning microscope. The amounts of intracellularly taken up sugars increased concomitantly with the formation of endocytotic vesicles depending on the time of incubation in parallel with a gradual increase of freezing tolerance. However, the endocytotic vesicles and their contents were retained not only after prolonged incubation after maximum freezing tolerance had been achieved but also after recovery of these tissue cells to isotonic conditions or after freeze-thawing. These results suggest that although sugars are intracellularly taken up by endocytotic vesicles, they might be sequestered within vesicles, casting doubt on their protective role to the plasma membranes as a main site of freezing injury. The pretreatment with 1 mM p-chloromercuribenzenesulfonic acid (PCMBS), an inhibitor of sugar transport, reduced the amounts of intracellular sugar uptake without affecting the formation of endocytotic vesicles, suggesting that sugars were, at least partly, taken up by sugar transporters. In the pretreatment with PCMBS, the freezing tolerance of incubated tissues with sugar solutions was significantly reduced, although addition of PCMBS per se did not affect survival. These results suggest that sugars taken up by sugar transporters, rather than sugars taken up by endocytotic vesicles, are mainly responsible for the increased freezing tolerance of cabbage petioles and asparagus shoot apexes. Furthermore, we aimed to study the occurrence of fluid-phase endocytosis with LYCH in an isotonic condition. Our results indicated that uptake of LYCH by fluid-phase endocytotic vesicles was not detected microscopically in isotonic condition, although LYCH was spectrofluorimetrically taken up in isotonic condition. Spectrofluorimetric uptake of LYCH was inhibited by addition of probenecid, an anion transport inhibitor. These results suggest that in cabbage petioles and asparagus shoot apexes, LYCH is taken up by anion transport but not by fluid-phase endocytosis in isotonic condition, and uptake of LYCH by fluid-phase endocytosis is restricted to occur only in hypertonic condition. PMID- 11732315 TI - Immunolocalization of alkaloids and X-ray microanalysis of elements in lupin seeds. AB - Immunolocalization of alkaloids in lupin seeds (Lupinus spp.) has been performed by cryofixation and conventional methods. Alkaloids were localized in the protein bodies of the cotyledon cells. Some immunogold particles in the walls of these cells were also observed. There were no differences in the sites of localization between the two mentioned methods. X-ray microanalysis of elements showed the presence of P, Mg, S, and K in the protein bodies of cotyledon cells in lupin seeds. The role of K+ in alkaloids transport is discussed. PMID- 11732316 TI - Botrytis cinerea induces the formation of free radicals in fruits of Capsicum annuum at positions remote from the site of infection. AB - Free radical adducts of the spin trap alpha-(4-pyridyl-1-oxide)-N-tert butylnitrone have been observed by electron paramagnetic resonance spectroscopy in detached fruits of Capsicum annuum investigated 5 days after infection with Botrytis cinerea. The spectra of these adducts were at a maximum within the soft rot lesion, but they could also be detected at distances up to 50 mm from the edge of the lesion in samples following main vascular bundles. At distances greater than 40 mm, the spectrum of the ascorbate radical was also seen, and at greater distances from the lesion it was the only radical detected. With samples taken from parenchyma tissue adjacent to the vascular bundles there was little adduct formation and the ascorbate radical could be detected, albeit with reduced intensity compared to healthy tissue, at distances as small as 10 mm from the edge of the lesion. This observation of chemical changes at considerable distances from the infected tissue is in contrast to previous observations on the behaviour of other markers of oxidative stress (e.g., 4-hydroxynonenal, malondialdehyde, single-peak free radical, and Fe(III) (g = 4.27) electron paramagnetic resonance signals), where their levels decreased rapidly outside of the soft rot. PMID- 11732314 TI - Plant calcium signaling and monitoring: pros and cons and recent experimental approaches. AB - This review focusses on Ca(2+)-mediated plant cell signaling and optical methods for in vivo [Ca2+] monitoring and imaging in plants. The cytosolic free calcium concentration has long been considered the central cellular key in plants. However, more and more data are turning up that critically question this view. Conflicting arguments show that there are still many open questions. One conclusion is that the cytosolic free Ca2+ concentration is just one of many cellular network parameters orchestrating complex cellular signaling. Novel experimental strategies which unveil interference of cellular parameters and communication of transduction pathways are required to understand this network. To date only optical methods are able to provide both kinetic and spatial information about cellular key parameters simultaneously. Focussing on calcium there are currently three classes of calcium indicators employed (i.e., chemical fluorescent dyes, luminescent indicators, and green-fluorescent-protein-based indicators). Properties and capabilities as well as advantages and disadvantages of these indicators when used in plant systems are discussed. Finally, general experimental strategies are mentioned which are able to answer open questions raised here. PMID- 11732317 TI - Smooth muscle proteins as intracellular components of the chromatophores of the Antarctic fishes Pagothenia borchgrevinki and Trematomus bernacchii (Nototheniidae). AB - Melanophores, xanthophores, and iridophores from the skins of the two Antarctic fish species Pagothenia borchgrevinki and Trematomus bernacchii were tested immunocytochemically for the presence of a variety of muscle proteins. Actin, myosin, and calmodulin, not surprisingly, were confirmed for all three chromatophore types of the two fishes, but the presence of caldesmon and calponin, both characteristic proteins of smooth muscle fibers, represents a new discovery. It is not known at this stage whether these proteins occur also in the chromatophores of other fishes and are not restricted to Antarctic species. Since, however, motility control of particles in fish chromatophores and the regulation of smooth muscle tension both involve the sympathetic nervous system, the presence of similar target proteins should not come as a surprise. The fact that none of the chromatophores tested positive for troponin shows that there is no close relationship between pigment cells and striated muscle. The lack of alpha-actinin in iridophores, but its presence in melanophores and xanthrophores, is thought to be a reflection of the considerably greater pigment translocations within the latter two types of chromatophore cells. PMID- 11732318 TI - Dynamic changes in the frequency and architecture of plasmodesmata during the sink-source transition in tobacco leaves. AB - The sink-source transition in tobacco leaves was studied noninvasively using transgenic plants expressing the green-fluorescent protein (GFP) under control of the Arabidopsis thaliana SUC2 promoter, and also by imaging transgenic plants that constitutively expressed a tobacco mosaic virus movement protein (MP) fused to GFP (MP-GFP). The sink-source transition was measured on intact leaves and progressed basipetally at rates of up to 600 microns/h. The transition was most rapid on the largest sink leaves. However, leaf size was a poor indicator of the current position of the sink-source transition. A quantitative study of plasmodesmatal frequencies revealed the loss of enormous numbers of simple plasmodemata during the sink-source transition. In contrast, branched plasmodesmata increased in frequency during the sink-source transition, particularly between periclinal cell walls of the spongy mesophyll. The progression of plasmodesmal branching, as mapped by the labelling of plasmodesmata with MP-GFP fusion, occurred asynchronously in different cell layers, commencing in trichomes and appearing lastly in periclinal cell walls of the palisade layer. It appears that dividing cells retain simple plasmodesmata for longer periods than nondividing cells. The rapid conversion of simple to branched plasmodesmata is discussed in relation to the capacity for macromolecular trafficking in developing leaf tissues. PMID- 11732319 TI - Ultrastructure, distribution, and transmission of endosymbionts in the whitefly Aleurochiton aceris Modeer (Insecta, Hemiptera, Aleyrodinea). AB - The body of the whitefly Aleurochiton aceris contains specialized cells, termed mycetocytes, that enclose endosymbiotic microorganisms. The endosymbionts are transmitted from one generation to the next transovarially. In contrast to other insects, in whiteflies whole intact mycetocytes migrate into the ovaries, traverse the follicular epithelium, and reach the oocyte surface (i.e., perivitellin space). The migration of mycetocytes begins in the last instar, called puparium, from which imagines emerge. During this stage the cytoplasm of mycetocytes is tightly packed with pleomorphic bacteria and less numerous coccoid microorganisms. In adult females the mycetocytes gather extracellularly in the depression of the vitellarial oocyte. Till the end of oogenesis neither pleomorphic nor coccoid microorganisms are released from mycetocytes into the oocyte. PMID- 11732320 TI - Isolation and quantification of the heat shock protein 90 alpha and beta isoforms from rat liver. AB - Heat shock protein 90 (Hsp90) is an abundant cytosolic protein. In higher eukaryotes two isoforms of Hsp90 exist, Hsp90 alpha and Hsp90 beta. Hsp90 was purified from rat liver and after sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a double band at about 90 kDa. The two bands were separated and identified as the Hsp90 alpha and Hsp90 beta isoforms. There was no entry in the protein databases for the Hsp90 alpha isoform from rat. Furthermore, the ratio of the two Hsp90 isoforms was determined. PMID- 11732322 TI - Acclimative changes in root epidermal cell fate in response to Fe and P deficiency: a specific role for auxin? AB - Root hair formation and the development of transfer cells in the rhizodermis was investigated in various existing auxin-related mutants of Arabidopsis thaliana and in the tomato mutant diageotropica. Wild-type Arabidopsis plants showed increased formation of root hairs when the seedlings were cultivated in Fe- or P free medium. These extranumerary hairs were located in normal positions and in positions normally occupied by nonhair cells, e.g., over periclinal walls of underlying cortical cells. Defects in auxin transport or reduced auxin sensitivity inhibited the formation of root hairs in response to Fe deficiency completely but did only partly affect initiation and elongation of hairs in P deficient roots. Application of the ethylene precursor 1-aminocyclopropane-1 carboxylic acid or the auxin analog 2,4-dichlorophenoxyacetic acid did not rescue the phenotype of the auxin-resistant axr2 mutant under control and Fe-deficient conditions, indicating that functional AXR2 product is required for translating the Fe deficiency signal into the formation of extra hairs. The development of extra hairs in axr2 roots under P-replete conditions was not affected by auxin antagonists, suggesting that this process is independent of auxin signaling. In roots of tomato, growth under Fe-deficient conditions induced the formation of transfer cells in the root epidermis. Transfer cell frequency was enhanced by application of 2,4-dichlorophenoxyacetic acid but was not inhibited by the auxin transport inhibitor N-1-naphthylphthalamic acid. In the diageotropica mutant, which displays reduced sensitivity to auxin, transfer cells appeared to develop in both Fe-sufficient and Fe-deficient roots. Similar to the wild type, no reduction in transfer cell frequency was observed after application of the above auxin transport inhibitor. These data suggest that auxin has no primary function in inducing transfer cell development; the formation of transfer cells, however, appears to be affected by the hormonal balance of the plants. PMID- 11732321 TI - Evidence that soilborne wheat mosaic virus moves long distance through the xylem in wheat. AB - Soilborne wheat mosaic virus (SBWMV) is a member of the genus Furovirus of plant viruses. SBWMV is transmitted to wheat roots by the plasmodiophorid vector Polymyxa graminis. Experiments were conducted to determine the path for SBWMV transport from roots to leaves. The results of immunogold labeling suggest that SBWMV enters and moves long distance through the xylem. SBWMV may enter primary xylem elements before cell death occurs and then move upward in the plant after the xylem has matured into hollow vessels. There is also evidence for lateral movement between adjacent xylem vessels. PMID- 11732323 TI - Membrane orientation of droplets prepared from Chara corallina internodal cells. AB - It is generally accepted that the membrane surrounding droplets from characean cells originates from the tonoplast, but there is some uncertainty regarding droplet membrane sidedness. This issue was addressed directly by combining two different droplet isolation methods and the patch clamp technique. Neutral red accumulation was used to demonstrate the presence of H(+)-transport over the membrane and to predict membrane orientation. Two types of droplet populations with differently oriented membranes could be formed in an iso-osmotic bath solution. Cytoplasmic droplets (cytosolic side of the tonoplast inside) contained cytoplasm, while the second type of droplet population contained vacuolar sap (vacuolar droplets, vacuolar side of the tonoplast inside). Smaller vesicels also appeared inside the droplets, with an apparently inversely oriented membrane. Confocal laser scanning microscopy indirectly demonstrated that, at least with one of the droplet isolation methods, the plasma membrane entirely remains in the internodal cell after intracellular perfusion. Both types of droplet populations allowed the formation of excised patches and single-channel measurements by the patch clamp technique. Properties of anion channels in the tonoplast could be used to prove the predicted membrane orientation, knowing that Ca2+ can only activate these channels from the cytosolic side. These results provide useful data for studies addressing ligand-binding, block and modulation, organization and interaction of proteins within the membrane or with other regulatory factors, where it is important to control membrane orientation. PMID- 11732324 TI - Direct interaction between glyoxysomes and lipid bodies in cotyledons of the Arabidopsis thaliana ped1 mutant. AB - During germination and subsequent growth of fatty seeds, higher plants obtain energy from the glyconeogenic pathway in which fatty acids are converted to succinate in glyoxysomes, which contain enzymes for fatty acid beta-oxidation and the glyoxylate cycle. The Arabidopsis thaliana ped1 gene encodes a 3-ketoacyl-CoA thiolase (EC 2.3.1.16) involved in fatty acid beta-oxidation. The ped1 mutant shows normal germination and seedling growth under white light. However, etiolated cotyledons of the ped1 mutant grow poorly in the dark and have small cotyledons. To elucidate the mechanisms of lipid degradation during germination in the ped1 mutant, we examined the morphology of the ped1 mutant. The glyoxysomes in etiolated cotyledons of the ped1 mutant appeared abnormal, having tubular structures that contained many vesicles. Electron microscopic analysis revealed that the tubular structures in glyoxysomes are derived from invagination of the glyoxysomal membrane. By immunoelectron microscopic analysis, acyl-CoA synthetase (EC 6.2.1.3), which was located on the membrane of glyoxysomes in wild type plants, was located on the membranes of the tubular structures in the glyoxysomes in the ped1 mutant. These invagination sites were always in contact with lipid bodies. The tubular structure had many vesicles containing substances with the same electron density as those in the lipid bodies. From these results, we propose a model in which there is a direct mechanism of transporting lipids from the lipid bodies to glyoxysomes during fatty acid beta-oxidation. PMID- 11732326 TI - Plasma membrane redox systems. PMID- 11732325 TI - Production of a recombinant human basic fibroblast growth factor with a collagen binding domain. AB - Basic fibroblast growth factor (bFGF) is a potent in vitro mitogen for capillary endothelial cells, stimulates angiogenesis in vivo, and may participate in tissue repair. Basic FGF is found in abundance in tissues such as brain, kidney, and cartilage. This study reports the expression, purification, and renaturation of a biologically active human basic fibroblast growth factor fusion protein (hbFGF F1) from Escherichia coli. A prokaryotic expression vector was engineered to produce a tripartite fusion protein consisting of a purification tag, a protease sensitive linker and collagen binding domain, and a cDNA sequence encoding the active fragment of hbFGF. The expressed hbFGF-F1 and hbFGF-F2 (it contains the collagen binding domain), located in inclusion bodies, were solubilized with 6 M guanidine-HCl and renatured by a glutathione redox system and protracted dialysis under various experimental conditions. The purification of the recombinant proteins was achieved by binding the His-tag of the fusion protein on a nickel nitrilotriacetic acid metal chelate column. The biological activity of the recombinant growth factor was demonstrated by its ability to stimulate proliferation of human vein endothelial cells, monitored by [3H]thymidine incorporation, where commercial recombinant human bFGF (rhbFGF) served as a positive control. Purified rhbFGF-F1 and rhbFGF-F2 constructs exhibited proliferative activity comparable to commercial rhbFGF. The high-affinity binding was demonstrated by the binding of [3H]collagen to the rhbFGF-F2 protein immobilized on a Ni-nitrilotriacetic acid column. The rhbFGF-F2 fusion protein bound to collagen-coated surfaces with high affinity. Taken together, these results demonstrate that biologically active rhbFGF fusion proteins can be recovered from transformed bacteria by oxidative refolding; thus, providing a means for their high-yield production, purification, and renaturation from microorganisms. Furthermore, we demonstrate that the auxiliary collagen binding domain effectively targets the recombinant growth factor to type I collagen. These studies advance the technology necessary to generate large quantities of targeted bFGF fusion proteins for specific biomedical applications. PMID- 11732327 TI - Reactive oxygen intermediates involved in cellular regulation. AB - Reactive oxygen intermediates (ROIs) in low concentration, as released permanently by nonphagocytic cells, possess important functions in inter- and intracellular signalling. They lead to alterations in the phosphorylation pattern followed by gene activation, including the expression of proto-oncogenes. Redox sensitive sites in membrane molecules may trigger adhesion and chemotaxis or open ion channels and activate transport processes across the cytoplasma membrane. ROIs shift the ratio of cyclic GMP to cyclic AMP giving signals to proliferation and differentiation processes. Senescence, apoptosis, and cell death can also be modulated by ROIs, depending on concentration and cell type. PMID- 11732329 TI - Role of extracellular peroxidase in the superoxide production by wheat root cells. AB - Extracellular peroxidase has been shown to contribute to superoxide production in wounded wheat (Triticum aestivum L. cv. Ljuba) root cells. The superoxide synthesizing system of root cells was considerably inhibited by KCN and NaN3 and activated by MnCl2 and H2O2. Treatment of roots with salicylic acid and a range of di- and tri-carbonic acids (malic, citric, malonic, fumaric, and succinic acids) stimulated superoxide production in both root cells and extracellular solution. The H2O2-stimulated superoxide production in the extracellular solution was much higher when roots were preincubated with salicylic or succinic acid. Exogenous acids enhanced peroxidase activity in the extracellular solution. Pretreatment of root cells with the detergents trypsin and sodium dodecyl sulfate had similar effects on the peroxidase activity. Significant inhibition of both superoxide production and peroxidase activity by diphenylene iodonium suggests that the specificity of the latter as an inhibitor of NADPH oxidase is doubtful. Results obtained indicate that extra-cellular peroxidase is involved in the superoxide production in wheat root cells. The mobile form of peroxidase can be readily secreted to the apoplastic solution and serve as an emergency enzyme involved in plant wound response. PMID- 11732328 TI - Superoxide generation of phagocytes and nonphagocytic cells. AB - Phagocytes release high amounts of reactive oxygen intermediates upon contact with appropriate stimuli into the environment as an important function in the immune defence against infectious agents. On the other hand nonphagocytic cells release low amounts upon stimulation, which have important functions in the inter and intracellular signal transduction. Both systems represent a class of isoenzymes making a specific regulation possible. Fibroblasts, hepatocytes, and HeLa cells possess a superoxide system which shows higher activity with NADH than NADPH. The enzyme was purified to homogeneity in an active form, showed a molecular mass of 84 kDa and possessed a flavin and low-potential cytochrome b558. PMID- 11732331 TI - Ascorbate-independent electron transfer between cytochrome b561 and a 27 kDa ascorbate peroxidase of bean hypocotyls. AB - Cytochrome b561 (cyt b561) is a trans-membrane cytochrome probably ubiquitous in plant cells. In vitro, it is readily reduced by ascorbate or by juglonol, which in plasma membrane (PM) preparations from plant tissues is efficiently produced by a PM-associated NAD(P)H:quinone reductase activity. In bean hypocotyl PM, juglonol-reduced cyt b561 was not oxidized by hydrogen peroxide alone, but hydrogen peroxide led to complete oxidation of the cytochrome in the presence of a peroxidase found in apoplastic extracts of bean hypocotyls. This peroxidase active on cyt b561 was purified from the apoplastic extract and identified as an ascorbate peroxidase of the cytosolic type. The identification was based on several grounds, including the ascorbate peroxidase activity (albeit labile), the apparent molecular mass of the subunit of 27 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, the dimeric native structure, the typical spectral properties of a heme-containing peroxidase, and an N-terminal sequence strongly conserved with cytosolic ascorbate peroxidases of plants. Cyt b561 used in the experiments was purified from bean hypocotyl PM and juglonol was enzymatically produced by recombinant NAD(P)H:quinone reductase. It is shown that NADPH, NAD(P)H:quinone reductase, juglone, cyt b561, the peroxidase interacting with cyt b561, and H2O2, in this order, constitute an artificial electron transfer chain in which cyt b561 is indirectly reduced by NADPH and indirectly oxidized by H2O2. PMID- 11732330 TI - Cellular redox state and activating protein-1 are involved in ascorbate effect on calcitriol-induced differentiation. AB - Ascorbate has been related to the differentiation of several mesenchymal cells including haematopoietic cells. We have previously demonstrated that ascorbate enhances the activity of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) on monocytic differentiation of HL-60 cells. Here, we show that ascorbate-mediated modification of cellular redox state and AP-1 (activating protein-1) DNA binding during early phases are related to the enhancing effect of ascorbate on differentiation. Ascorbate, but not its fully oxidized form, dehydroascorbate, or an ascorbate analogue with a low rate of oxidation, ascorbate-2-phosphate, enhanced the differentiation induced by 1 alpha,25(OH)2D3, modified cytosolic reactive oxygen species levels and mitochondrial redox potential (delta psi m), and modulated AP-1 DNA binding in HL-60 cells. Ascorbate itself increased AP-1 binding to DNA in noninduced cells, whereas it inhibited AP-1 binding in 1 alpha,25(OH)2D3-induced cells. However, ascorbate increased the mRNA levels of c jun, junB, and c-fos in 1 alpha,25(OH)2D3-induced cells. Taken together, these results suggest that the enhancing effect of ascorbate on HL-60 differentiation induced by 1 alpha, 25(OH)2D3 is related to its effect on the cellular redox state and the modulation of AP-1 activity. PMID- 11732332 TI - Different diurnal cycles of expression of two nitrate reductase transcripts in tobacco roots. AB - In roots and leaves of tobacco (Nicotiana tabacum cv. Samsun) three functional transcripts (3.6 kb, 3.1 kb, and 1.8 kb) were found to at least partly represent nitrate reductase mRNA. With specific probes for the transcripts of the different domains of nitrate reductase it was shown that the smallest transcript was shortened in the region coding for the flavin adenine dinucleotide domain and might be the transcript coding for plasma-membrane-bound nitrate reductase. The expression of the 3.1 kb and 1.8 kb transcripts in roots was differently regulated during the day-night cycle with the maximum amount of the 3.1 kb transcript in the middle and of the 1.8 kb transcript at the end of the light period. PMID- 11732333 TI - Pathogenic infection and the oxidative defences in plant apoplast. AB - The structural and functional continuum of the plant apoplast is the first site of contact with a pathogen and plays a crucial role in initiation and coordination of many defence responses. In this paper, we present an overview of the involvement of the plant apoplast in plant-pathogen interactions. The process of infection of French bean (Phaseolus vulgaris L.) plants by Colletotrichum lindemuthianum is analysed. The ultrastructural features of plant defence responses to fungal infection are then compared with those observed in plants or cell suspensions treated with various elicitors. Changes in cell walls and in whole plant cells responding to infection seem to be highly similar in all systems used. Model systems of French bean and white lupin (Lupinus albus L.) are then utilised to provide some biochemical characteristics of oxidative reactions in the apoplast evoked by elicitor treatment. The species specificity of various mechanisms generating reactive oxygen species is discussed, and some details of pH-dependent H2O2-generating activity of peroxidases are demonstrated. As its exocellular nature is an important feature of the oxidative burst, the major consequence of this event, i.e., the oxidative cross-linking of wall components during the papilla formation and strengthening of the walls, is analysed. Finally, the possible involvement of other wall-associated and developmentally regulated H2O2-generating mechanisms, like amine and oxalate oxidases, in plant defence is demonstrated. It is concluded that under stress conditions, such apoplastic mechanisms might be employed to increase plants' chances of survival. PMID- 11732334 TI - Inhibition of trans-membrane hexacyanoferrate III reductase activity and proton secretion of maize (Zea mays L.) roots by thenoyltrifluoroacetone. AB - Intact plants can reduce external oxidants by an appearingly trans-membrane electron transport. In vivo an increase in net medium acidification accompanies the reduction of the apoplastic substrate. Up to now, several NAD(P)H dehydrogenases, b-type cytochromes, and a phylloquinone have been identified and partially purified from plant plasma membranes. The occurrence of a quinone in the plasma membrane of maize roots supports the hypothetical model of a proton transferring redox system, i.e., an electron transport chain with a quinone as mobile electron and proton carrier. In the present study the trans-membrane electron transport system of intact maize (Zea mays L.) roots was investigated. Flow-through and ionostat systems have been used to estimate the electron and proton transport activity of this material. Application of 4,4,4-trifluoro-1-(2 thienyl)-butane-1,3-dione (thenoyltrifluoroacetone) inhibited the reduction of ferricyanide in the incubation solution of intact maize roots up to 70%. This inhibition could not be washed off by rinsing the roots with fresh incubation medium. The acidification of the medium induced after ferricyanide application was inhibited to about 62%. The effects of thenoyltrifluoroacetone on proton fluxes in the absence of ferricyanide have been characterized in a pH-stat system. The net medium acidification by maize roots was inhibited up to 75% by thenoyltrifluoroacetone in the absence of ferricyanide, while dicumarol inhibited net acidification completely. The inhibition of H(+)-ATPase activity was estimated with plasma membrane vesicles isolated by phase partitioning and treated with 0.05% (w/v) Brij 58. ATP-dependent proton gradients and Pi release were measured after preincubation with the effectors. The proton pumping activity by those plasma membrane vesicles was inhibited by dicumarol (53.6%) and thenoyltrifluoroacctone (77.8%), while the release of Pi was unaffected by both inhibitors. PMID- 11732335 TI - How superoxide radical damages the cell. AB - Superoxide is considered to be poorly reactive, and cell damage has been attributed to HO. generated via the Haber-Weiss reaction. The function of O2- in this reaction is only to reduce Fe3+ to Fe2+. In vivo, however, superoxide could not out-compete cellular reductants such as glutathione, NADPH, and ascorbate, which makes the observed O2- toxicity rather puzzling. Little attention has been paid to the idea that, irrespective of its poor chemical reactivity, superoxide might be capable of interacting directly with specific intracellular targets; and that even the Haber-Weiss reaction might be a consequence of such direct interactions. This paper summarizes latest data that support the concept of such a mechanism. PMID- 11732336 TI - NADPH oxidase subunit gp91phox: a proton pathway. AB - The generation of superoxide by the NADPH oxidase is an electrogenic process resulting in a rapid depolarisation of the membrane potential of the cell. The efflux of H+ ions through an arachidonate-activatable, Zn(2+)-inhibitable H+ pathway accompanies the efflux of electrons and provides the necessary charge compensation. Inhibition of H+ flux leads to inhibition of superoxide generation. The protein gp91phox, a transmembrane component of the NADPH oxidase, was demonstrated to be capable of acting as the NADPH oxidase-associated H+ channel in a stable CHO cell line, CHO91. The N-terminal 230 amino acids contain all that is required for the protein to form an H+ channel and specifically histidine 115 is important to the ability of gp91phox to conduct H+ ions. The recording of outward currents from CHO91 cells, in the whole-cell configuration, demonstrated that gp91phox is also capable of functioning as a voltage-gated H+ conductance pathway. The similarity in properties between voltage-elicited outward currents, from both wild type and the mutations, and the arachidonate-activated H+ flux strongly suggests that these H+ pathways are one in the same. Among the recently identified homologues of gp91phox only NOH-1S has so far been demonstrated to also act as an H+ conductance pathway. PMID- 11732337 TI - Thioredoxin and glutathione system of malaria parasite Plasmodium falciparum. AB - Plasmodium falciparum is the causative agent of malaria tropica. Due to the increasing resistance towards the commonly used plasmodicidal drugs there is an urgent need to identify and assess new targets for the chemotherapeutic intervention of parasite development in the human host. It is established that P. falciparum-infected erythrocytes are vulnerable to oxidative stress, and therefore efficient antioxidative systems are required to ensure parasite development within the host cell. The thioredoxin and glutathione redox systems represent two powerful means to detoxify reactive oxygen species and this article summarizes some of the recent work which has led to a better understanding of these systems in the parasite and will help to assess them as potential targets for the development of new chemotherapeutics of malaria. PMID- 11732338 TI - b-type cytochromes in plasma membranes of Phaseolus vulgaris hypocotyls, Arabidopsis thaliana leaves, and Zea mays roots. AB - The plasma membrane of higher plants contains more than one kind of b-type cytochromes. One of these has a high redox potential and can be fully reduced by ascorbate. This component, the cytochrome b561 (cyt b561), has its characteristic alpha-band absorbance close to 561 nm wavelength at room temperature. Cyt b561 was first isolated from etiolated bean hook plasma membranes by two consecutive anion exchange chromatography steps. During the first step performed at pH 8, cyt b561 did not bind to the anion exchange column, but other b-type cytochromes did. In the second step performed at pH 9.9, cyt b561 was bound to the column and was eluted from the column at an ionic strength of about 100 mM KCl. However, when the same protocol was applied to the solubilized plasma membrane proteins from Arabidopsis thaliana leaves and maize roots, the ascorbate-reducible cyt b561 bound already to the first anion exchange column at pH 8 and was eluted also at an ionic strength of about 100 mM KCl. Other b-type cytochromes than the ascorbate-reducible cyt b561 from the plasma membranes of Arabidopsis leaves and maize roots showed similar chromatographic characteristics to that of bean hypocotyls. These results demonstrate particular differences in the chromatographic behavior of cyt b561 from different sources. PMID- 11732339 TI - Effect of dichlorophenolindophenol, dichlorophenolindophenol-sulfonate, and cytochrome c on redox capacity and simultaneous net H+/K+ fluxes in aeroponically grown seedling roots of sunflower (Helianthus annuus L.): new evidence for a plasma membrane CN(-)-resistant redox chain. AB - Excised roots from axenically grown sunflower seedlings reduced or oxidized exogenously added 2,6-dichlorophenolindophenol (DCIP), DCIP-sulfonate (DCIP-S), and cytochrome c, and affected simultaneous H+/K+ net fluxes. Experiments were performed with nonpretreated "living" and CN(-)-pretreated "poisoned" roots (control and CN(-)-roots). CN(-)-roots showed no H+/K+ net flux activity but still affected the redox state of the compounds tested. The hydrophobic electron acceptor DCIP decreased the rate of H+ efflux in control roots with extension of the maximum rate and optimal pH ranges, then the total net H+ efflux ([symbol: see text]H+) equalled that of the roots without DCIP. The simultaneously measured K+ influx rate was first inhibited, then inverted into efflux, and finally influx recovered to low rates. This effect could not be due to uptake of the negatively charged DCIP, but due to the lower H+ efflux and the transmembrane electron efflux caused by DCIP, which would depolarize the membrane and open outward K+ channels. The different H+ efflux kinetics characteristics, together with the small but significant DCIP reduction by CN(-)-roots were taken as evidence that an alternative CN(-)-resistant redox chain in the plasma membrane was involved in DCIP reduction. The hydrophilic electron acceptor DCIP-S enhanced both H+ and K+ flux rates by control roots. DCIP-S was not reduced, but slightly oxidized by control roots, after a lag, while CN(-)-roots did not significantly oxidize or reduce DCIP-S. Perhaps the hydrophobic DCIP could have access to and drain electrons from an intermediate carrier deep inside the membrane, to which the hydrophilic DCIP-S could not penetrate. Also cytochrome c enhanced [symbol: see text]H+ and [symbol: see text]K+, consistent with the involvement of the CN(-) resistant redox chain. Control roots did not reduce but oxidize cytochrome c after a 15 min lag, and CN(-)-roots doubled the rate of cytochrome c oxidation without any lag. NADH in the medium spontaneously reduced cytochrome c, but control or CN(-)-roots oxidized cytochrome c, despite of the presence of NADH. In this case CN(-)-roots were less efficient, while control roots doubled the rate of cytochrome c oxidation by CN(-)-roots, after a 10 min lag in which cytochrome c was reduced at the same rate as the medium plus NADH did. CN(-)-roots seemed to have a fully activated CN(-)-resistant branch. The described effects on K+ flux were consistent with the current hypothesis that redox compounds changed the electric membrane potential (de- or hyperpolarization), which induces the opening of voltage-gated in- or outward K+ channels. PMID- 11732340 TI - Changes in plasma membrane fluidity of corn (Zea mays L.) roots after Brij 58 treatment. AB - The detergent Brij 58 has been introduced to reverse plasma membrane (PM) vesicles from the right-side-out to the inside-out form. The aim of the present work was to investigate the effect of Brij 58 on the formation of an ATP dependent proton gradient and on the fluidity of the lipid phase of PM vesicles. PMs of corn (Zea mays L.) roots were isolated by phase-partitioning. The fluidity of PMs was estimated by measurement of fluorescence polarization with 1-(4 trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) and 1,6-diphenyl 1,3,5-hexatriene (DPH). The PMs of corn roots were relatively rigid. The hydrophobic part of the lipid bilayer was more fluid than the hydrophilic part. After intercalation of Brij 58 into the lipid bilayer the membrane fluidity changed in a concentration-dependent manner. Treatment with the detergent Brij 58 increased the degree of fluorescence polarization for TMA-DPH, while it decreased it for DPH. This effect was saturated at a detergent-to-protein ratio of 1:4 for both fluorescence probes. Although the biophysical characteristics of the membrane were changed after Brij 58 treatment, the formation of ATP-dependent proton gradients could still be measured with those vesicles. The generation of an ATP-dependent proton gradient with Brij 58-treated PM vesicles suggests that the detergent treatment indeed turned the originally right-side-out vesicles to sealed inside-out vesicles. The limits of the effect caused by Brij 58 in the context of PM enzyme activities are discussed. PMID- 11732341 TI - Interaction between electron transport at the plasma membrane and nitrate uptake by maize (Zea mays L.) roots. AB - In the present study nitrate uptake by maize (Zea mays L.) roots was investigated in the presence or absence of ferricyanide (hexacyanoferrate III) or dicumarol. Nitrate uptake caused an alkalization of the medium. Nitrate uptake of intact maize seedlings was inhibited by ferricyanide while the effect of dicumarol was not very pronounced. Nitrite was not detected in the incubation medium, neither with dicumarol-treated nor with control plants after application of 100 microM nitrate to the incubation solution. In a second set of experiments interactions between nitrate and ferricyanide were investigated in vivo and in vitro. Nitrate (1 or 3 mM) did neither influence ferricyanide reductase activity of intact maize roots nor NADH-ferricyanide oxidoreductase activity of isolated plasma membranes. Nitrate reductase activity of plasma-membrane-enriched fractions was slightly stimulated by 25 microM dicumarol but was not altered by 100 microM dicumarol, while NADH-ferricyanide oxidoreductase activity was inhibited in the presence of dicumarol. These data suggest that plasma-membrane-bound standard-ferricyanide reductase and nitrate reductase activities of maize roots may be different. A possible regulation of nitrate uptake by plasmalemma redox activity, as proposed by other groups, is discussed. PMID- 11732342 TI - Higher-plant plasma membrane cytochrome b561: a protein in search of a function. AB - During the past twenty years evidence has accumulated on the presence of a specific high-potential, ascorbate-reducible b-type cytochrome in the plasma membrane (PM) of higher plants. This cytochrome is named cytochrome b561 (cyt b561) according to the wavelength maximum of its alpha-band in the reduced form. More recent evidence suggests that this protein is homologous to a b-type cytochrome present in chromaffin granules of animal cells. The plant and animal cytochromes share a number of strikingly similar features, including the high redox potential, the ascorbate reducibility, and most importantly the capacity to transport electrons across the membrane they are located in. The PM cyt b561 is found in all plant species and in a variety of tissues tested so far. It thus appears to be a ubiquitous electron transport component of the PM. The cytochromes b561 probably constitute a novel class of transmembrane electron transport proteins present in a large variety of eukaryotic cells. Of particular interest is the recent discovery of a number of plant genes that show striking homologies to the genes coding for the mammalian cytochromes b561. A number of highly relevant structural features, including hydrophobic domains, heme ligation sites, and possible ascorbate and monodehydroascorbate binding sites are almost perfectly conserved in all these proteins. At the same time the plant gene products show interesting differences related to their specific location at the PM, such as potentially N-linked glycosylation sites. It is also clear that at least in several plants cyt b561 is represented by a multigene family. The current paper presents the first overview focusing exclusively on the plant PM cyt b561, compares it to the animal cyt b561, and discusses the possible physiological function of these proteins in plants. PMID- 11732343 TI - The existence and significance of redox-cycling ubiquinone in lysosomes. AB - Ubiquinone is inhomogeneously distributed in subcellular biomembranes. Apart from mitochondria, where ubiquinone was demonstrated to exert bioenergetic and pathophysiological functions, unusually high levels of ubiquinone were also reported to exist in Golgi vesicles and lysosomes. In lysosomes the interior differs from other organelles by the low pH value which is important not only to arrest proteins but also to ensure optimal activity of proteases. Since redox cycling of ubiquinone is associated with the acceptance and release of protons, we assumed that ubiquinone is a part of a redox chain contributing to unilateral proton distribution. A similar function of ubiquinone was earlier reported to exist in Golgi vesicles. Support for the involvement of ubiquinone in a presumed couple of redox carriers came from our observation that almost 70% of total lysosomal ubiquinone was in the divalently reduced state. Further reduction was seen in the presence of external NADH. Analysis of the components involved in the transfer of reducing equivalents from cytosolic NADH to ubiquinone revealed the existence of a flavin adenine dinucleotide-containing NADH dehydrogenase. The latter was found to reduce ubiquinone by means of a b-type cytochrome. Proton translocation into the interior was linked to the activity of the novel lysosomal redox chain. Oxygen was found to be the terminal electron acceptor thereby also regulating acidification of the lysosomal matrix. The role of the proton-pumping redox chain has to be elucidated. PMID- 11732345 TI - The cyber response to bioterrorism. PMID- 11732344 TI - The ascorbate: ascorbate free radical oxidoreductase from the erythrocyte membrane is not cytochrome b561. AB - Erythrocytes contain a plasma membrane redox system that can reduce extracellular ascorbate radicals by using intracellular ascorbate as an electron donor. In this study, the hypothesis was tested that cytochrome b561 was a component of this system. Spectroscopic analysis of erythrocyte membrane preparations revealed the presence of cytochrome b5 and hemoglobin but also of a cytochrome with properties similar to cytochrome b561, reducible by ascorbate and insensitive to CO. The presence of cytochrome b561 was studied further by reverse transcriptase-PCR analysis of erythrocyte progenitor cells, reticulocytes. However, no cytochrome b561 mRNA could be found. These results were corroborated by Western blot analysis with an anti-cytochrome b561 serum. No cytochrome b561 protein could be detected in extracts of erythrocyte membranes. It is therefore concluded that erythrocytes do not contain cytochrome b561 in their membranes. The possible involvement of other b-cytochromes in ascorbate-ascorbate free radical oxidoreductase activity is discussed. PMID- 11732346 TI - Covenants not to compete--what physicians need to know. PMID- 11732347 TI - Now is the time for HIPAA compliance. AB - HIPAA privacy regulations are complex and extensive. All those affected, including physician practices, must now begin the process of education and planning necessary to ensure compliance with these regulations by April 14, 2003. Starting now will ease the burden of implementation in the future. This article addresses specific issues discussed in the final privacy rules as they pertain to the establishment of compliance with these rules, especially regarding physician practices. PMID- 11732348 TI - Potential HIPAA provider benefits. PMID- 11732349 TI - First steps in HIPAA compliance for the physician's practice. PMID- 11732350 TI - Just another hemoptysis or a fluke? AB - Hemopytsis is commonly encountered in the daily practice of the pulmonary physician. Younger patients with normal chest x-rays frequently have acute or chronic bronchial disease accounting for their complaint. Occasionally parasitic disease is described as an unusual cause for a patient presenting with hemoptysis. Although pulmonary paragonimiasis is unusual in this country, because of the rapid growth in travel as well as immigration, physicians will need to be aware of this disorder. PMID- 11732351 TI - The Task Force on E. coli O157 Final Report: the view from here. PMID- 11732352 TI - On being prepared: the Influenza Pandemic Plan. PMID- 11732353 TI - Use of hygiene advice and active immunisation to control an outbreak of hepatitis A. AB - A community outbreak of at least 16 cases of hepatitis A is described. Salivary antibody testing of 126 children within a school in that community showed only one case of undiagnosed previous infection. Three other children had received hepatitis A vaccine previously, and this was reflected in their antibody titres. For the schools serving the community control measures included advice on good hygiene (supervised handwashing and additional cleaning in the schools). All close contacts of cases (household and childminding contacts and all staff in one school) were given active immunisation, though two subsequently developed hepatitis A, 17 and 27 days after vaccination. The use of human normal immunoglobulin (HNIG) was fully discussed with contacts, but was declined in all cases. The low prevalence of previous infection found in the schoolchildren implies that, in future, few adults will be immune. Public concern about blood products after the recent Department of Health decision to stop using British sourced plasma may be a factor in limiting the uptake of HNIG, especially as there is now a perceived safer alternative, namely hepatitis A vaccine. Lack of evidence of spread of disease within the schools involved demonstrates that good hygiene, especially handwashing, remains the most important element in the control of hepatitis A. PMID- 11732354 TI - An outbreak of hepatitis A among young men associated with having sex in public venues. AB - An increase in hepatitis A virus (HAV) infection was noted among young men in the former Thames regions during 1997. A retrospective case-control study, using a standardised questionnaire at interview, was conducted in the area most affected (London and East Sussex) to investigate the hypothesis that this increase was mainly among homosexual men and to establish the risk factors associated with transmission. Forty-eight cases and 161 controls completed questionnaires. Forty one cases (85%) described their sexuality as homosexual (p < 0.0001). Cases were more likely than controls to have eaten shellfish (Odds Ratio (OR) 2.4; 95% Confidence Interval (CI) 1.16, 5.04) during the two months before onset of illness. Cases had more sexual partners (p = 0.015), and more casual sexual partners (p = 0.007) than controls. Cases were more likely to have had sex in a gay sauna (OR 3.5; 95% CI 1.53, 8.30), or in a gay club, pub or disco (OR 2.9; 95 CI 1.29, 6.63) than controls. After adjusting for confounding factors, cases were more likely to have eaten shellfish (adjusted [adj] OR 3.0; 95% CI 1.33, 6.59) and to have had sex in a gay sauna (adj OR 3.9; 95% CI 1.42, 10.59). Public health messages need to inform homosexual men about recognised risk factors such as eating shellfish and travel abroad to endemic areas, as well as sexual risks. Homosexual men can benefit from hepatitis A vaccine. We would suggest that in an outbreak situation men who have multiple anonymous partners and have sex in public venues should be targeted as a priority for health education and immunisation. PMID- 11732355 TI - General outbreaks of infectious intestinal disease linked with poultry, England and Wales, 1992-1999. AB - Between 1992 and 1999, 1426 foodborne general outbreaks of infectious intestinal disease (IID) were reported to the Public Health Laboratory Service Communicable Disease Surveillance Centre. A fifth were associated with the consumption of poultry. Chicken was implicated in almost three quarters of these outbreaks, turkey in over a fifth and duck in 2% of outbreaks. The organisms most frequently reported were Salmonella (30% of outbreaks), Clostridium perfringens (21%) and Campylobacter (6%). Over 7000 people were affected, with 258 hospital admissions and 17 deaths. During the summer, outbreaks were mainly of salmonellosis and attributed to the consumption of chicken. In December, C. perfringens and turkey were the organism and vehicle most often implicated. Most outbreaks occurred on commercial catering premises (56%) or in private houses (21%). The highlight of this surveillance period was the fall in outbreaks of salmonellosis linked with poultry products, probably due, at least in part, to the vaccination of poultry flocks. PMID- 11732356 TI - Can calls to NHS Direct be used for syndromic surveillance? AB - This study assessed whether NHS Direct could be a useful source of surveillance data for communicable diseases, using influenza as a pilot condition. Data on the weekly total number of calls and the number from people reporting influenza-like symptoms to three pilot NHS Direct sites were collected between November 1999 and March 2000. NHS Direct data were compared with routinely available influenza surveillance data. The NHS Direct call rate peaked at 331 per 100,000 population in week 52 of 1999. The percentage of calls for 'influenza-like illness' (one site) peaked at 15% during week 51. Information about weekly call numbers to NHS Direct could be produced in a timely way. It was not clear whether the observed peak in calls reflected a true increase in influenza or whether it was the result of an increase in calls over the Christmas/Millennium holiday period due to more difficulty in accessing other services. The ability to assess the proportion of calls made directly by, or on behalf of, each age group will be of vital importance in interpreting seasonal respiratory disease. PMID- 11732357 TI - Improving influenza immunisation coverage in 2000-2001: a baseline survey, review of the evidence and sharing of best practice. AB - In May 2000 a new government target of 70% uptake in people aged 65 and over was introduced for influenza immunisation, with a minimum of 60% uptake in 2000-2001. A postal survey of influenza immunisation coverage in over 75 year olds during 1999-2000 was undertaken in our district's general practices. The evidence on interventions which improve coverage was compared to local practice. General Practitioners and their Primary Care Groups/Trust received feedback on their performance, and best practice was shared. The findings were used to inform the immunisation campaign for 2000-01. The survey response rate was 74%. The district coverage was 50%, ranging from 7% to 97% between practices. Practices achieving high coverage rates combined good patient identification with personalised patient invitation and well-organised clinics. The work that went on in the district achieved an overall influenza immunisation coverage of 62% in 2000-01. PMID- 11732358 TI - Surveillance of listeriosis in England and Wales, 1995-1999. AB - A total of 543 cases of listerIosis were ascertained by the Public Health Laboratory Service Communicable Disease Surveillance Centre (PHLS CDSC) in residents of England and Wales with onset of illness between 1st January 1995 and 31st December 1999. Of 452 non-pregnancy associated cases, 326 (72%) had major medical conditions, 140 (31%) had received immuno-suppressive therapy and 97 (21%) were aged 60 years or over. Serovar 4b predominated in both pregnancy and non-pregnancy associated cases. Of ninety-one pregnancy associated cases, five had underlying medical conditions. Foetal death or early neonatal death was reported in 24 (26%) of the cases giving a mean perinatal mortality rate of 7.5 per million conceptions per year. Two episodes of neonatal cross infection occurred. Two non-pregnancy associated cases in 1999 were linked by a common food; all other cases were unlinked. No trend was observed from 1995 to 1999. PMID- 11732359 TI - Campylobacter reporting at its peak year of 1998: don't count your chickens yet. AB - Infections due to Campylobacter spp. are the most commonly reported bacterial cause of human gastrointestinal illness, and reports of campylobacteriosis in the UK and other temperate countries have been increasing. An all-time peak of 58,059 cases of campylobacter infectious intestinal disease (IID) were reported to the Communicable Disease Surveillance Centre in 1998. An analysis of the age, gender and seasonal and geographical distribution of cases reported in 1998 is presented here, together with population-based estimates of reported incidence. The frequency of reports was highest in late Spring. Reported incidence was highest in males under 1 year of age (214/100,000 population), with a secondary peak in adults aged 25 to 34 years (140/100,000 population). Reported incidence was higher in males than females for all age groups. The highest reported incidence occurred in the South West region (140/100,000 population), while the lowest was observed in London (54/100,000 population). Campylobacter is a major cause of human gastrointestinal illness. Work towards its control must be a priority if the burden of human infectious intestinal disease is to be reduced. PMID- 11732360 TI - The 'real-world' impact of improved diagnostic techniques for Chlamydia trachomatis infection in Glasgow. AB - In April 1997, the main chlamydia laboratory in Glasgow introduced ligase chain reaction (LCR) as its standard diagnostic test. The diagnostic effectiveness and health economic impact of introduction of LCR testing was assessed. Between April 1996 to March 2000, results of all chlamydia detection tests on genital specimens sent from general practitioners and the two main sexual healthcare providers (Genitourinary Medicine and Family Planning services) were reviewed. A preliminary economic assessment, inclusive of staff, reagents, consumables and laboratory overheads was conducted. Overall, testing activity increased four and a half times between 1996-97 and 1999-2000; the proportionate rise was greatest in general practice. Although chlamydia testing in both genders increased over the review period, testing activity rose disproportionately in women (59%, compared with a 31% increase in men). The overall Chlamydia trachomatis detection rate rose from 4.8% in 1996-97 to 7.8% in 1999-2000. Following introduction of LCR testing, an estimated additional 331 men and 844 women were diagnosed during the study period. The cost per additional diagnosis made was estimated at 162 Pounds for men and 263 Pounds for women. Substantial health gains are likely to be achieved, at both an individual and public health level, as a result of introduction of LCR testing for genital chlamydial infection. PMID- 11732361 TI - Survey of Staphylococcus aureus contamination in a hospital's spa and hydrotherapy pools. AB - Twenty-seven percent of water samples taken from one spa and two hydrotherapy pools in one Welsh hospital over three months contained Staphylococcus aureus. Four per cent of samples were deemed unacceptable because they contained coliforms, E. coli or Pseudomonas aeruginosa. Aerobic colony counts varied between samples but no counts were > 100 cfu/ml. Disinfection and pH records indicated no significant problems with pool maintenance over the sampling period, and bather loads were generally low. Some samples positive for S. aureus were found after lengthy periods of pool inactivity, indicating persistence of the organism. PMID- 11732362 TI - Clinical and microbiological features of suspect sporadic food poisoning cases presenting to an accident and emergency department. AB - During a five-month period all patients presenting to an inner-city accident and emergency department with suspected food poisoning were assessed for relevant clinical and microbiological features. There were 102 patients in the study period (0.3% of all attendances). Fifty-two stool samples were processed and 11 were culture positive (21% of those sampled, 11% of all cases). The two commonest organisms were Campylobacter (6) and Salmonella (4). A logistic regression analysis showed that, of the continuous variables, a higher temperature (odds ratio [OR] 2.72, 95% confidence interval [CI] 1.36 to 5.46) was predictive of stool culture positivity, and age was negatively predictive (OR 0.88, 95% CI 0.78 to 0.98). Of the dichotomous variables only a history of fever (OR 3.94, 95% CI 1.07 to 14.5) was predictive of stool culture positivity. The presence of nausea (OR 0.11, 95% CI 0.03 to 0.43) or vomiting (OR 0.22, 95% CI 0.06 to 0.80), and the absence of abdominal tenderness, had a negative predictive value for stool culture positivity. No patient without a history of abdominal pain and diarrhoea had a positive stool culture. PMID- 11732364 TI - The importance of protecting injecting drug users against hepatitis A. PMID- 11732363 TI - Guidelines for the control of hepatitis A virus infection. AB - The PHLS Advisory Committee on Vaccination and Immunisation, following a review of the evidence on control measures for preventing hepatitis A virus (HAV) infection and widespread consultation, has prepared the following guidelines. They include a description of the current epidemiology of HAV infection in England and Wales, where most individuals are now susceptible to HAV. HAV infection is uncommon, with around 1000 infections notified per year in England and Wales. Clusters occur in families and in settings where potential for faecal/oral spread is high, e.g. day care centres, nurseries, primary schools. Larger outbreaks have been recorded in men who have sex with men and injecting drug users. Personal hygiene remains the cornerstone of measures for preventing HAV infection and its spread. Those with haemophilia, hepatitis B or C virus infection or liver cirrhosis, intravenous drug users and men who have sex with men should be offered HAV vaccination as a preventive measure. HAV vaccine should be used for preventing secondary cases and outbreaks provided that patients are informed that the latest date the vaccine is most likely to be effective in preventing disease in contacts is probably 7 days from onset of illness in the primary case. Human normal immunoglobulin (HNIG) should be offered in addition or in preference to vaccine for contacts who are more than 7 days from onset of illness in the primary case, and for those at risk of adverse outcome of HAV infection. Individuals at particular risk of an adverse outcome to infection include those more than 50 years old, with liver cirrhosis of any cause, or with pre-existing hepatitis B or C virus infection. HAV vaccine should be used to prevent infection for travellers to countries where HAV infection is a risk. HNIG is no longer indicated for travellers. Children travelling to such countries should be offered vaccine from 5 years and consideration should be given to vaccinating those aged 1-4 years. PMID- 11732365 TI - Selecting aptamers--the way to find a fit. PMID- 11732366 TI - Laboratory partnership with the Medical Devices Agency. AB - To improve the quality of the information and advice provided to the Health Service, the Medical Devices Agency (MDA) is actively seeking to increase the number of reports from hospital laboratories. The Medical Devices Agency relies heavily on laboratory reports of problems with in vitro diagnostic medical devices to investigate and take action where necessary. Laboratories are encouraged to report all suspected adverse incidents to the MDA. PMID- 11732367 TI - Clinical waste management. AB - Every producer of controlled waste in the UK has a duty of care under the Environmental Protection Act 1990 to dispose of the waste in a manner which takes appropriate care and consideration to ensure that it does not cause harm to human health or pollute the environment. This article looks at what is meant by 'duty of care' to ensure clinical waste produced by laboratories or in a health care setting is disposed of in the correct manner. PMID- 11732368 TI - [Geriatric wards in a psychiatric and a general hospital: a comparison of problems leading to admission of the patient]. AB - The aim of this investigation was the comparison and description of actual problems of patients admitted to a geriatric ward of a mental hospital and patients admitted to the geriatric ward of a general hospital. The study was conducted in the geriatric unit of the Vincent van Gogh Institute of Psychiatry, in Venray and the geriatric unit in St. Maartens Gasthuis in Venio, both in the Netherlands. The design was retrospective. Data were obtained from discharge letters. The relevant diagnoses and the aetiology of the problems of all patients admitted in 1994 were categorized and compared by means of classification systems and models (medical, synergistic and causal chain model). Approximately 80 percent of the patients admitted to the geriatric unit of the mental hospital turned out to have a physical diagnosis relevant to the problems and almost every patient had a relevant psychiatric diagnosis. Life events and relationship problems played an important part. More than half of the patients admitted to the geriatric unit of the general hospital had a relevant psychiatric diagnosis and almost every patient had a physical diagnosis relevant to the problems. Mostly the aetiology of the problems of the patients of the geriatric unit of the mental hospital can be described by means of the medical model. The aetiology of the problems of the patients of the geriatric unit of the general hospital can mainly be described by the synergistic model. The causal chain model is seldom found. An event is more frequently found in the geriatric unit of the mental hospital. Often the referring Doctor could not point out adequately the underlying cause of the problems. We conclude that the problems of the patients, admitted to the geriatric unit of a mental and of a general hospital, differ substantially and in both units the problems are complex, in terms of diagnoses, aetiology and/or treatment. PMID- 11732369 TI - [Three geriatric wards of a psychiatric hospital (GAPZ): a comparison of patient admissions]. AB - The aim of this investigation was the description and comparison of problems of patients admitted to three geriatric wards of a mental hospital. The study took place in the geriatric units at the Vincent van Gogh Institute of psychiatry in Venray, the Netherlands and was retrospective. Data obtained from discharge letters were analysed and the relevant diagnoses and the aetiology of the problems of all patients admitted in 1994 were categorised and compared. This was done bij means of classification systems and models of Fried et al. The problems of patients admitted to the geriatric unit of a mental hospital very frequently appeared to be combined problems of a psychiatric, somatic and/or social nature. In only 58% there was a simple connection between the cause(s) and the current problems. Differences between the three wards were small and non significant. In conclusion, the problems of the patients, admitted to the geriatric units of a mental hospital, did not differ significantly between the three wards. Furthermore, they are complex in terms of diagnosis, aetiology and/or treatment. PMID- 11732370 TI - [Validity of the tympanic infrared thermometer in geriatric inpatients]. AB - The purpose of this study was to assess the agreement between rectal temperature and infrared emission detection (IRED). The specificity and sensitivity, the positive predictive value and the negative predictive value were calculated. It was a prospective, observational and unblinded study. Thermoscan-pro-1 was used to collect pairs of data in 41 patients, all hospitalised on an acute geriatric ward during a 4-month period. 41% of the patients were febrile. The 'bias' between rectal and tympanic measurement (left ear) was -0.64 degree C (d) and the standard deviation 0.58 degree C (s). The '95% Confidence Interval' for the bias was -0.82 degree C and -0.46 degree C. The '95% limits of agreement' between the rectal measurement and the tympanic measurement (left ear) was -1.78 degrees C and 0.50 degree C. Regression analysis taught us that bias was increasing with increasing temperature. If we take into account one measurement with the IRED ear thermometry, the sensitivity was 41%, the specificity 83%. The positive predictive value was 64% en the negative predictive value 67%. We can conclude that the IRED ear thermometry is not a valuable alternative for the rectal, digital thermometry in the elderly, frail and febrile hospital inpatients. PMID- 11732371 TI - [Sensory stimulation: new apparatus for demented men]. AB - In order to study the feasibility of new sensorily stimulating material for elderly demented men, their behaviour was observed while the material was handed over to them. The newly developed materials consisted of five different constructions, i.e. a mirrorbox, a simple hand-operated stroboscope, a spring operated ball for ringing a bell, two rotating wooden cogwheels, and a box having a sandpaper bottom of different grains causing a tactile contrast. In their verbal and nonverbal responses the demented men showed clear signs of interest in the constructions and the materials. The seven participating men showed much interest in materials made of wood and metal, and in the working of the constructions. The different constructions and materials could be combined into one complete 'snoezelproduct'. PMID- 11732373 TI - [Dying among the younger aged, what is known? Aging of short duration also needs to be studied]. PMID- 11732372 TI - [Improvement of vision in nursing home patients through eyeglasses]. AB - Adequate refraction correction may contribute to the quality of life of elderly persons who will be less dependent on care in daily life and will be less prone to fall. In nearly 20% of 102 nursing home residents binocular visual acuity improved with at least one line on the Snellen Chart by adjustment of refractive correction. PMID- 11732374 TI - On a wish and a prayer: healing through distant intentionality. PMID- 11732375 TI - Integrative health and healing: "looking in" to "left out" medicine. PMID- 11732376 TI - Is there any evidence? PMID- 11732377 TI - Stroke and complementary medicine. PMID- 11732378 TI - Integrative medicine approach to obesity and diabetes. PMID- 11732379 TI - Complementary and alternative healing in paradise: the state of the state. PMID- 11732380 TI - Researching nutrition and breast cancer. PMID- 11732381 TI - About prizes, awards and grants. PMID- 11732383 TI - Transition between anesthesia and post-operative analgesia: relevance of intra operative administration of analgesics. AB - The rapid awakening without residual analgesic effect seen with the new "fast-in fast-out" anesthetic agents forces us to anticipate post-operative pain management. More then ever, "balanced analgesia" appears the key for successful transition between anesthesia and post-operative analgesia. This review focuses on practical aspects seeking to answer the following questions: which analgesics should be used?; how should they be given?; at what stage of the anesthetic time should they be injected? In the absence of a loco-regional technique, analgesics must be given before the end of surgery to obtain a quiet awakening. The transition in this delicate period may be facilitated by the intra-operative use of various adjuvant therapies such as ketamine, lidocaine infusion, clonidine, and magnesium, that reduce the needs for opioid and/or post-operative pain severity. All non-opioid analgesics (propacetamol, NSAIDs, tramadol) must be given according to their pharmacokinetic characteristics, indications and contraindications to ensure the required analgesic effect is effective at the time of awakening. If tramadol is not used, an initial dose of a long-acting opioid should be given 20-30 minutes before the end of surgery. Insufficient analgesia must be corrected by titrating an opioid intravenously before allowing the patient to control his/her analgesia using a PCA pump. PMID- 11732382 TI - Piroxicam versus tenoxicam in spine surgery: a placebo controlled study. AB - In double-blind trial 60 patients undergoing spine surgery were randomized to receive either placebo, tenoxicam 40 mg intravenously (i.v.), tenoxicam 40 mg intramuscularly (i.m.) or piroxicam 40 mg i.m., immediately following the induction of general anesthesia. As compared to placebo, the 24 hour morphine consumption was reduced in all groups. This reduction was only statistically significant (p = 0.023) in the i.v. group (21.7 +/- 11.27 versus 36.53 +/- 20.33 mg). Rest and dynamic pain scores were significantly lower in both tenoxicam groups but more consistently in the i.v. group. With piroxicam only rest pain scores at 24 hours were lower. Less urinary retention was noticed in the i.v. tenoxicam group. This study shows that, following spine surgery, i.v. tenoxicam induces a morphine sparing effect (41%) while offering lower rest and dynamic pain scores and a lower incidence of urinary retention. PMID- 11732385 TI - Miss-'n-mix and mimics. PMID- 11732384 TI - Liver and renal function after volatile induction and maintenance of anesthesia (VIMA) with sevoflurane versus TIVA with sufentanil-midazolam for CABG surgery. AB - We compared liver and renal function after volatile induction and maintenance of anesthesia (VIMA) with sevoflurane and minimal dose sufentanil versus total intravenous anesthesia (TIVA) with moderate dose of midazolam and sufentanil in patients undergoing CABG surgery. Eighty nine patients were studied retrospectively after VIMA (44 patients) or after TIVA (45 patients). Liver and renal function were measured before (T0), then 1 (T1), 2 (T2), 5 (T3) days and 6 weeks (T4) after the operation. Serum levels of aspartate aminotransferase (ASAT) and lactate dehydrogenase (LDH) increased in both groups at T1, T2 and T3 and the highest levels were at T1. Levels of total bilirubin (TBil) increased at T1 only in the TIVA group. Levels of g-glutamyl transpeptidase (GGT) and alanine aminotransferase (ALAT), increased in both groups at T3. Serum levels of creatinine (Cr) were high in both groups on T1 and T2. Blood urea nitrogen (BUN) was high in both group at T2, and T3 with the peak levels in T2. Six weeks after the operation all liver and renal functions were normal in both groups. We concluded that VIMA with sevoflurane during cardiac surgery has no untoward effects on liver or renal functions. The transient reversible elevation was comparable in the VIMA and TIVA groups which was most probably due to the effect of the operation itself. PMID- 11732386 TI - Recovery of neuromuscular block in morbidly obese patients following an infusion of rocuronium. PMID- 11732387 TI - Comparative study of the TOF-ratio measured by the ParaGraph versus the TOF Guard, with and without thumb repositioning. PMID- 11732388 TI - Comparison of the influence of desflurane nitrous oxide anesthesia to propofol nitrous oxide anesthesia on rocuronium bromide induced neuromuscular block. PMID- 11732389 TI - [Three of four diabetics die of coronary heart disease. What permits sweet heart to beat longer?]. PMID- 11732390 TI - [Moderate alcohol consumption. Already 10 grams daily raise the carcinoma risk by 30%]. PMID- 11732391 TI - [Ligament, meniscus and cartilage injuries. Knee repairs always more refined]. PMID- 11732392 TI - [Cycle disorders, polydipsia, decline of vision. Is it the fault of the hypophysis?]. AB - Apart from partial or complete insufficiency, further diseases of the pituitary with clinical impact are those associated with hormone overproduction such as acromegaly, Cushing's disease and prolactinoma. The cardinal symptom of pituitary insufficiency or disruption of the female cycle, loss of libido or sexual potency reflecting a lesion of the gonadotrophic axis. Hyperprolactinemia also results in a loss of gonadotrophic function. Macroadenomas of the pituitary gland often give rise to visual field defects, so that an ophthalmological work-up is a must. With the exception of hyperprolactinemia, measurement of basic hormone levels is not always sufficient to detect pituitary diseases. For the establishment of hormone deficiency, therefore, stimulation tests, and for detection of hypersecretion suppression tests, are carried out. Owing to the increasing utilization of cranial MRI, incidental detection of intrasellar tumors, the so-called incidentalomas, is becoming ever more common, and these lesions require at least an endocrinological investigation. PMID- 11732394 TI - [Medical students and their health. Afflictions of the Cand. med]. PMID- 11732393 TI - [Therapy of pituitary diseases. What can be achieved with medication and hormones?]. AB - Pharmacotherapy is available only for pituitary adenomas producing growth hormone and prolactin (acromegaly, hyperprolactinemia). While dopamine agonists and somatostatin analogues are usually administered to acromegaly patients only after unsuccessful surgery, dopamine agonists are the treatment of choice in patients with prolactinomas. Replacement treatment in patients with insufficiency of the anterior lobe of the hypophysis is oriented to the extent of the deficits. Theoretically, replacement treatment can be applied for any functional loss. Of essential importance is the replacement of lost adrenal cortical function by the regular administration of physiological doses of hydrocortisone, and of lost thyroid function by the administration of L-thyroxine. In high-stress situations, for example, due to severe illness, the hydrocortisone dose must be increased significantly, and if necessary given parenterally. Gonadal function is replaced either with gonadal steroid hormones or gonadotropin or GnRH in patients wishing to have children. In adults, replacement of growth hormone is also indicated to decrease cardiovascular risk. This, however is very expensive and requires s.c. injections. PMID- 11732395 TI - [Does resection of pituitary adenoma and radiotherapy cure the patient's symptoms?]. AB - Today, pituitary adenomas are readily amenable to surgical treatment, and complication rates are low. With the exception of prolactin-producing adenomas, transsphenoidal or transcranial surgery is usually the treatment of choice. Indications for surgery are considered to be visual field deficits, functional disorders of the optomotor cranial nerves, or hypo-/hyperfunction of the pituitary. If treatment leads to normalization of the secretory dynamics, the long-term prognosis is favorable. If residual tumor or persistent overproduction of pituitary hormones is detected postoperatively, radiotherapy is applied. Endoscopic surgical procedures, intra-operative imaging and peri-operative determination of hormone levels enable the radicality of tumor removal to be further increased. PMID- 11732397 TI - [Schroder intervenes in drug savings package. Sparing for the industry, more restrictions for us physicians?]. PMID- 11732396 TI - [Emergencies in the practice. Hazards of "mix syringe"]. PMID- 11732398 TI - [Drug manufacturer on the savings package of Ulla Schmidt. "Grip in the moth box of planned economics"]. PMID- 11732399 TI - [For the first time a health insurance company publishes drug data. Surprise: the thriftiest colleagues are in the East]. PMID- 11732400 TI - [Metabolism control in overweight type 2 diabetics. High metformin dosage brings more]. PMID- 11732401 TI - [Monoclonal antibodies as adjuvant breast cancer therapy. Tumor cells in continuing sleep?]. PMID- 11732402 TI - [Respiratory tract inflammations in children. Passive smoking increases the risk]. PMID- 11732403 TI - [Interference in the serotoninergic and noradrenergic system. Faster out of depression]. PMID- 11732404 TI - [Osteoporosis therapy with bisphosphonates. Once weekly is enough]. PMID- 11732405 TI - [PEG-interferon alfa 2a against hepatitis C. Better as the standard therapy]. PMID- 11732406 TI - [A new antibiotic against respiratory tract infections. More power against pneumococci]. PMID- 11732407 TI - [Tooth implantation. Robots prevent nerve injuries]. PMID- 11732408 TI - [60% of patients with coronary disease receive antilipemic drugs. Nevertheless cholesterol often is too high]. PMID- 11732409 TI - [Thus to avoid complications of diabetes. To combine early]. PMID- 11732411 TI - A conceptual framework for the economic analysis of factors influencing decision making of small-scale farmers in animal health management. AB - The delivery of veterinary services in most of sub-Saharan Africa has undergone substantial changes, with the private sector gaining increasing recognition as an alternative to state provision. Given this policy shift, the authors argue that a better understanding of the behaviour and decision-making processes of small scale farmers in animal health management is urgently required, to guide policy decisions regarding the delivery of animal health services. Whether the involvement of the private sector will improve overall efficiency in the delivery of veterinary services will depend greatly on the demand response of livestock producers who must make decisions about the health of livestock. The authors briefly review the decision-making process in small-scale farming systems, the economic nature of animal diseases and disease control and the models that have been used to guide resource allocation for disease control. To gain an improved understanding of the behaviour and decision-making processes of small-scale farmers, the authors propose a conceptual model including variables that relate to characteristics specific to small-scale farmers and farms, economic factors, institutional setting and biophysical factors. Two possible approaches are proposed for the economic analysis. Agricultural household modelling allows the derivation and testing of hypotheses regarding the demand elasticities for veterinary services, while qualitative choice models are better suited to the analysis of determinants behind the choices of farmers. The authors conclude that an urgent need exists for empirical research in this area. PMID- 11732410 TI - The expected economic impact of selected exotic diseases on the pig industry of Australia. AB - The authors assess the expected economic impact of three exotic diseases on the pig industry of Australia. An integrated epidemiological/economic approach was used to assess the effects of classical swine fever, Nipah virus and porcine reproductive and respiratory syndrome. Scenarios involving either an epidemic event, in which the outbreaks were confined to selected regions and were eradicated, or an endemic situation, in which the diseases became established in Australia, were studied. Based only on loss of sales and disposal costs, epidemics resulted in regional losses in income of the order of AUS$10 million AUS$30 million (16%-37%) depending on disease and region. If any of these diseases became established, opportunity losses in gross national pig income of 5%-11% per year would occur, with classical swine fever the most serious of the three diseases. Establishment of any of the diseases would lead to rapid structural change in the pig industry, with concomitant social and economic dislocation in regional Australia. PMID- 11732412 TI - Eradication of foot and mouth disease in Japan. AB - An outbreak of foot and mouth disease (FMD) was recorded in Japan in the spring of 2000, the first for ninety-two years. Between 25 March and 11 May, four farms were infected. However, the disease was eradicated without resorting to vaccination, through a campaign of culling, movement control of cloven-hoofed animals in areas surrounding infected premises, and intensive clinical and serological surveillance. Japan regained FMD-free status by the end of September 2000. The authors describe the nature of the outbreaks, the eradication measures implemented, and the clinical and serological surveillance methods used. The possible sources of infection are also examined. Finally, the direct and indirect economic losses are presented. PMID- 11732413 TI - Importation of beef from countries infected with foot and mouth disease: a review of risk mitigation measures. AB - Risk mitigation measures to reduce the risks associated with importing beef from countries affected by foot and mouth disease (FMD) consist of controls at the farm of origin, inspection of slaughterhouses and maturation and deboning of carcasses. This assessment evaluates the effect of these measures on the mitigation of the risks presented by meat from cattle with FMD, for each of the different stages of the disease. The four disease stages considered are the incubation period, the period of clinical signs, convalescence and the carrier stage. Efficient animal health systems, disease surveillance, and ante-mortem and post-mortem inspection of all cattle effectively reduce the risk of FMD transmission from cattle slaughtered during the period of clinical signs or convalescence. These measures fail if the cattle are slaughtered during the incubation period, because of the absence of clinical signs. Cattle in this stage of the infection are likely to be viraemic, with FMD virus present in the skeletal muscles. Maturation of the carcasses of viraemic cattle reduces the risk of virus presence in the beef. In addition, deboning and removal of the principal lymph nodes and large blood vessels eliminate a source of FMD contamination of the beef. However, the slaughter of viraemic cattle creates an additional hazard of gross environmental viral contamination of the slaughterhouse facilities. Therefore, the maturation process may create a false sense of security, and the emphasis should instead be placed on disease surveillance within the infected zone and on the farms of origin, to prevent the slaughter of herds that are incubating FMD. Cattle slaughtered during the carrier stage do not pose a risk for the international beef trade. PMID- 11732414 TI - A qualitative assessment of the risk of introducing foot and mouth disease into Russia and Europe from Georgia, Armenia and Azerbaijan. AB - A qualitative risk assessment was performed to evaluate the risk of introducing foot and mouth disease (FMD) virus into Russia and the rest of Europe from the countries of Transcaucasia (Armenia, Azerbaijan and Georgia). The assessment was based on data collected during a three-week mission to these countries by the Food and Agriculture Organization, the European Union and the Office International des Epizooties in March 1999. Due to the strong involvement of the local veterinary services, much information was obtained. Although the data were not sufficient to allow a quantitative risk assessment to be performed, the investigation served as a useful initial approach, prior to undertaking a quantitative risk assessment. The risk of FMD virus infection is a function of two elements, namely: the probability of the hazard (virus infection) occurring, and the magnitude of the consequences. The probability of the hazard occurring is the product of the probability of entry of the virus and the probability of exposure to the virus. These elements were assessed using the following parameters: prevalence of infection; volume of trade; capacity of the virus to survive; and potential for infection. The magnitude of the consequences is derived from the probability of transmission and spread. Combining these parameters, the probability of occurrence of the hazard was rated as 'moderate'. Economic consequences of potential transmission and spread of FMD, in the local context, were rated as 'negligible'. As a result, the overall risk of introducing FMD virus into Russia and the rest of Europe from Trancaucasia was rated as 'low' at the time of evaluation. The method and results are presented to serve as a basis for further discussion. PMID- 11732415 TI - Using climate data to map the potential distribution of Culicoides imicola (Diptera: Ceratopogonidae) in Europe. AB - Culicoides imicola, a vector of bluetongue virus and African horse sickness virus, is principally Afro-Asian in distribution, but has recently been found in parts of Europe. A logistic regression model based on climate data (temperature, saturation deficit, rainfall and altitude) and the published distribution of C. imicola in Iberia was developed and then applied to other countries in Europe, to identify locations where C. imicola could become established. The model identified three temperature variables as significant determinants of the distribution of C. imicola in Iberia (minimum of the monthly minimum temperatures, maximum of the monthly maximum temperatures and number of months per year with a mean temperature > or = 12.5 degrees C). The model indicated that under current conditions, the distribution of C. imicola in Spain, Greece and Italy could be extended and the vector could potentially invade parts of Albania, Yugoslavia, Bosnia and Croatia. To simulate the effect of global warming, temperature values in the model were increased by 2 degrees C. Under these conditions, the potential spread of C. imicola in Europe would be even more extensive. PMID- 11732416 TI - Comparison of the efficacy of Brucella abortus strain RB51 and Brucella melitensis Rev. 1 live vaccines against experimental infection with Brucella melitensis in pregnant ewes. AB - To test the efficacy of rough Brucella strain vaccines in sheep, a vaccine recently developed in cattle (Brucella abortus strain RB51) was assessed in comparison with the conventional Rev. 1 vaccine. Forty-five ewes from twelve to fourteen months of age, from brucellosis-free flocks, were allotted to three groups of fifteen ewes each. Group one was vaccinated by the conjunctival route with 1.73 x 10(8) colony forming units (CFU) of Rev. 1 vaccine. Group two was vaccinated subcutaneously with 11 x 10(9) CFU of RB51 vaccine and group three was considered as a control. All sheep were challenged at two to three months of gestation with 5 x 10(7) CFU of virulent B. melitensis H38. Vaccination with RB51 vaccine did not result in the production of any antibodies against the O-side chain of lipopolysaccharide, as measured by conventional serological tests (Rose Bengal plate test and complement fixation test). Protection of sheep against abortion and excretion of virulent Brucella strain in vaginal fluid, aborted foetuses and/or non viable lambs at parturition and abortion was significantly lower than that afforded by Rev. 1 vaccine. The difference compared to the control group was not significant. Data from this study suggest that the RB51 vaccine used for cattle vaccination does not provide effective protection of sheep against abortion induced by B. melitensis. PMID- 11732417 TI - Development and field validation of an avidin-biotin enzyme-linked immunosorbent assay kit for bovine brucellosis. AB - The avidin-biotin enzyme-linked immunosorbent assay (A-B ELISA), for use in surveillance for bovine brucellosis in India was developed and calibrated using the indirect brucellosis ELISA kit of the International Atomic Energy Agency (IAEA) as a reference. The reagents used in the A-B ELISA were as follows: the smooth lipopolysaccharide of Brucella abortus strain 99 (antigen); biotinylated anti-bovine immunoglobulin G (detection antibody); avidin-horseradish peroxidase (conjugate); and O-phenylenediamine dihydrochloride (chromogen). The test results were interpreted using the IAEA software EDI version 2.1.1, which was modified for use in the A-B ELISA. The cut-off percentage positivity value was established using 500 brucellosis-positive and 500 brucellosis-negative serum samples, confirmed with reference to the sample data using the indirect ELISA kit. The overall specificity of A-B ELISA was 98.8% and overall sensitivity was 98.2%. Field validation of the A-B ELISA kit was undertaken in six laboratories in India. Screening of 7,040 cattle and 678 buffalo serum samples from 12 states revealed serological evidence of brucellosis in 8.7% of cattle and 10.2% of buffalo. This kit proved to be robust and performed with a similar sensitivity and specificity to the indirect ELISA. The kit can be supplied at a lower cost than current commercial ELISA kits. PMID- 11732418 TI - [Epidemiologic surveillance and prevention of brucellosis in ruminants: the example of the north African region and the Near East]. AB - The author reviews the general principles and different strategies recommended for the epidemiological surveillance and control of brucellosis in cattle and small ruminants, with particular reference to the region of North Africa and the Near East. Three strategic options are proposed, the choice of which depends on the real prevalence of the disease, the socio-economic context, the state of advancement of the animal health surveillance system and the policy set by the competent authorities. In heavily infected countries, gradual changeover is recommended from strategy A (systematic vaccination) to strategy B (selective vaccination), and eventually to strategy C (control measures), concurrent with the establishment of an adequate veterinary infrastructure, in particular for epidemiological surveillance and the control of animal movements. The author stresses the relevance for the majority of countries in the region in question of implementing the guidelines drawn up by the Food and Agriculture Organization, the World Health Organization and the Office International des Epizooties to control brucellosis in the Middle East. PMID- 11732419 TI - Field investigations of brucellosis in cattle and small ruminants in Syria, 1990 1996. AB - The authors present the epidemiological status of brucellosis in cattle and small ruminants in Syria from 1990 to 1996, based on laboratory findings at the Brucellosis Centre, Damascus. Initial investigations using the Rose Bengal plate test, the complement fixation test and a miniaturised variant of the slow agglutination test were conducted throughout the country in 1990 and 1991, revealing an overall herd seroprevalence rate of 3.14% in cattle herds and 2.94% in small ruminant flocks. Although partially biased by previous vaccination of young female cattle with S19 vaccine, these figures indicate that brucellosis in cattle is widespread, particularly in the urban governorates (provinces) of Damascus, Aleppo and Suwaydah. Brucellosis seroprevalence in sheep and goats was relatively high in the governorates of Damascus, Aleppo and Dara'a. The results of a second series of investigations, performed between 1992 and 1996, show that herd seroprevalence in cattle decreased steadily from 17.48% in 1992, to 2.59% in 1996, in the Government-owned farms, while seroprevalence increased in the private sector during the same period. The difference may be explained by the restriction of brucellosis vaccination to public farms (although this was far from systematic), combined with partial application of a 'test-and-slaughter' policy. In sheep and goats, brucellosis seroprevalence fluctuated in the two sectors, but was higher in the private sector where husbandry is principally extensive. Bacteriological investigations led to the isolation of Brucella melitensis biovars 2 and 3 in sheep and B. abortus biovar 9 in cattle. Although no specific methodology was employed, particularly with regard to sampling, this study is significant as the first international report of the distribution of brucellosis in Syria. Further, well-structured studies are required, the results of which could be used to plan an appropriate national control programme for brucellosis. PMID- 11732420 TI - First report of an outbreak of ovine septicaemic listeriosis in Saudi Arabia. AB - Although a case of human listeriosis has recently been reported in Saudi Arabia, the disease has not been reported in animals to date. The authors describe an outbreak of septicaemic listeriosis in sheep, which occurred during winter. Adult animals and pregnant ewes were principally affected, with a morbidity rate of 7.1% and a mortality rate of 2.4%; no abortions were recorded during the outbreak. Clinical signs included inappetence, weakness, fever, respiratory distress, keratoconjunctivitis and compulsive circling. Listeria monocytogenes was isolated in pure culture from affected sheep. Pathological findings indicated septicaemic listeriosis with encephalitis. Hygienic measures and antibiotics were successful in treating the rest of the flock. Aspects of the outbreak and public health implications are discussed. PMID- 11732421 TI - Immune response to Newcastle disease virus in broilers: a useful model for the assessment of detoxification of ervil seeds. AB - Ervil (Vicia ervilia) seeds are produced in the Mediterranean region and used as a source of protein for cattle and poultry. The methods used to assess the toxic effects of legume seeds in the feed of poultry include the observation of one or more parameters, including abnormal signs, weight gain, feed consumption, gall bladder weight, pancreas weight, pancreas proteolytic and amylase activity, haemolysis of red blood cells, liver weight, liver glutathione level, liver and plasma lipid levels, and plasma lipid peroxide levels. The authors describe the use of quantitative determination of the immune response to Newcastle disease virus (NDV) vaccine in broilers as a model that can be used to assess different detoxification treatments of ervil seeds. Broiler chicks fed differently-treated ervil, supplemented as 25% of the diet, at one to four weeks of age and vaccinated intraocularly with live NDV vaccine at eight days of age, showed different immune responses at three weeks post vaccination. Immunosuppression with regards to NDV was apparent in the group of birds raised on untreated ervil supplement, resulting in a mean immune response (titre) of 798.5. Five of the six different treatments of ervil seeds resulted in different degrees of rectification of the immunosuppression, with some broilers reaching a mean NDV immune titre of 2070.6, similar to that obtained in control broilers raised on a basal diet with no ervil seed supplement (mean NDV immune titre of 2333.8; P > 0.05). The five successful treatments of ervil (in increasing order of rectification of immunosuppression in broilers, with mean NDV titres in parentheses) were: ground soaked dried ervil (971.6), ground autoclaved dried ervil (1223.1), soaked autoclaved dried ervil (1273.1), soaked dried ervil (1340.0), and ground-soaked autoclaved dried ervil (2070.6). PMID- 11732422 TI - Study of gastro-intestinal helminths of scavenging chickens in four rural districts of Amhara region, Ethiopia. AB - A total of 267 rural scavenging chickens were examined from October 1998 to August 1999 in four woredas (districts) of the Amhara Region, Ethiopia. Of these chickens, 243 (91.01%) were found to harbour one to nine different helminth parasites and 24 (8.99%) were free of helminth parasites. A significant difference (P < 0.01) was found between the prevalence rates of helminth parasites in the different agro-ecological zones; the highest prevalence was observed in the lowland areas. This suggests that agro-ecology has a major influence on the distribution of helminth parasites. Nematodes recovered included Heterakis gallinarum (17.28%), Subulura brumpti (17.60%), Ascaridia galli (35.58%), Cheilospirura hamulosa (0.75%) and Dyspharynx spiralis (2.62%). The principal cestode species encountered were Raillietina echinobothrida (25.84%), Raillietina tetragona (45.69%), Raillietina cesticillus (5.62%), Amoebotaenia sphenoides (40.45%), Davainea proglottina (1.12%) and Choanotaenia infundibulum (4.49%). PMID- 11732423 TI - Antimicrobial resistance: an overview. AB - Increased antimicrobial resistance in bacteria that are important pathogens of humans, and spread of resistance from the closed environment of hospitals into open communities are increasingly perceived as a threat to public health. Any antimicrobial use, whether in humans, animals, plants or food processing technology, could lead to bacterial resistance. Use of antimicrobials in livestock production is suspected to significantly contribute to this phenomenon in species of bacteria which are common to humans and animals. Further research is required into the specific use conditions that govern the selection and dissemination of resistant bacteria. International travel and trade in animals and food increase the risks of antimicrobial resistance world-wide. Countries are considering import restrictions for products deemed a risk to public health. The Office International des Epizooties, a World Trade Organization reference organisation for the Agreement on the Application of Sanitary and Phytosanitary Measures, develops international standards on antimicrobial resistance which, as is the case for national measures, must be based on risk analysis. The scientific background and problems of resistance in human medicine are reviewed. Current knowledge, missing information and actions to be taken are identified. PMID- 11732424 TI - Antimicrobial resistance: risk analysis methodology for the potential impact on public health of antimicrobial resistant bacteria of animal origin. AB - The Ad hoc Group of experts on antimicrobial resistance, appointed by the Office International des Epizooties, has developed an objective, transparent and defensible risk analysis process, providing a valid basis for risk management decisions in respect to antimicrobial resistance. The components of risk analysis and of different possible approaches in risk assessment (qualitative, semiquantitative and quantitative) are defined. The Ad hoc Group recommended the following: an independent risk assessment based on scientific data; an iterative risk analysis process; a qualitative risk assessment systematically undertaken before considering a quantitative approach; the establishment of a risk assessment policy; and the availability of technical assistance for developing countries. PMID- 11732425 TI - Antimicrobial resistance: responsible and prudent use of antimicrobial agents in veterinary medicine. AB - A guideline on the responsible and prudent use of antimicrobials in animal husbandry has been developed by the Ad hoc Group of experts on antimicrobial resistance, created by the Office International des Epizooties. The objectives of responsible use are to maintain antibiotic efficacy, to avoid the dissemination of resistant bacteria or resistance determinants and to avoid the exposure of humans to resistance through food. The guideline attributes a central role to the competent authorities responsible for granting marketing authorizations for antimicrobial substances. Requirements before and after granting of marketing authorizations are defined. Important aspects include the control of the pharmaceutical product quality and the therapeutic efficacy, the assessment of the selection pressure, the protection of the environment, specific and non specific antimicrobial resistance surveillance. The guideline is also addressed to the veterinary pharmaceutical industry, veterinary practitioners, dispensing pharmacists and farmers. The respective roles and responsibilities of these groups are defined. PMID- 11732426 TI - Antimicrobial resistance: monitoring the quantities of antimicrobials used in animal husbandry. AB - This guideline, developed by the Office International des Epizooties for the monitoring of the quantities of antimicrobials used in animal husbandry, provides the methodology required to assess the amounts of antimicrobials used, to supply data to be used for risk analysis and to improve guidance on the appropriate use of antimicrobials. Information may be gathered from a number of sources, such as the competent authorities, industry and users. The usefulness of different types of information is discussed and recommendations are given on how to collect detailed information, each year, on the antimicrobial quantities used per class and active substance. Information should also be collected on the route of administration (oral and parenteral) and the animal species. PMID- 11732427 TI - Antimicrobial resistance: standardisation and harmonisation of laboratory methodologies for the detection and quantification of antimicrobial resistance. AB - The Ad hoc Group of experts on antimicrobial resistance of the Office International des Epizooties has developed a guideline on the standardisation and harmonisation of laboratory methodologies used for the detection and quantification of antimicrobial resistance. The existing methods (disk diffusion [including concentration gradient strips], agar dilution and broth dilution) are reviewed, including a comparison of their advantages and disadvantages. The definitions of resistance characteristics of bacteria (susceptible, intermediate and resistant) are addressed and the criteria for the establishment of breakpoints are discussed. Due consideration has to be given to these aspects in the interpretation and comparison of resistance monitoring or surveillance data. The use of validated laboratory methods and the establishment of quality assurance (internal and external) for microbiological laboratory work and the reporting of quantitative test results is recommended. Equivalence of different methods and laboratory test results is also recommended to be established by external proficiency testing, which should be achieved by the means of a reference laboratory system. This approach allows the comparison of test results obtained using different methods generated by laboratories in different countries. PMID- 11732428 TI - Antimicrobial resistance: harmonisation of national antimicrobial resistance monitoring and surveillance programmes in animals and in animal-derived food. AB - A guideline on the harmonisation of national antimicrobial resistance monitoring and surveillance programmes in animals and animal-derived foods has been developed by the Ad hoc Group of experts on antimicrobial resistance of the Office International des Epizooties. The objective of the guideline is to allow the generation of comparable data from various national surveillance and monitoring systems in order to compare the situations in different regions or countries and to consolidate results at the national, regional and international level. Definitions of surveillance and monitoring are provided. National systems should be able to detect the emergence of resistance, and to determine the prevalence of resistant bacteria. The resulting data should be used in the assessment of risks to public health and should contribute to the establishment of a risk management policy. Specific factors identified for harmonisation include the animal species, food commodities, sampling plans, bacterial species, antimicrobials to be tested, laboratory methods, data reporting, database structure and the structure of reports. PMID- 11732429 TI - [New medical assistance for reproduction in France: welcoming the embryo ]. PMID- 11732430 TI - [Obstetrical vacuum: a modern instrument]. AB - Contrary to the forceps, the vacuum extractor has clearly progressed in the last years. The use of the vacuum extractor increases in every developed countries, certainly because of an easier learning than forceps. Furthermore, maternal after effects of the delivery like sphincters injuries and anal incontinence seem to be less frequent with vacuum extractor than with forceps. For these reasons the American College of Gynecologists and Obstetricians (ACOG) recommend a large use and a priority teaching of this fetal extraction instrument. The technical aspects of use of the vacuum extractor are developed in this article, and personnel results are added as commentaries. PMID- 11732431 TI - [Acceptability and value of autoinjection of gonadotropins for ovarian stimulation in intrauterine insemination]. AB - OBJECTIVES: To evaluate the acceptability, the personal and economic benefit of subcutaneous self-injections of recombinant FSH within mono-ovulating stimulation for Intra Uterine Insemination (IUI). PATIENTS AND METHODS: Women aged < 42 years' old, enrolled for a series of three IIU associated with ovarian stimulation with FSH. All had an infertility > 2 years, at least one patent fallopian tube, and normal FSH and estradiol levels at day three of the cycle. In the male partner, the migration test yielded > 10(6) mobile spermatozoa with survivals > 10% after 24 h. Once entered in the study, the patients (with or without their partners) were informed and shown how to administer the injection. After each cycle, they filled up a questionnaire evaluating the training and the possible difficulties they had with their infertility treatment. RESULTS: Forty women were treated. 103 cycles were achieved (including 40 first cycles, 35 second cycles and 28 third cycles) 99 IUI were performed and ten pregnancies were obtained. Four cycles were cancelled: Premature fall of estradiol levels (n = 2), multiple pregnancy risk (n = 1) and spontaneous ovulation (n = 1). Three pregnancies occurred spontaneously between treatment cycles. All the patients appreciate to perform subcutaneous self-injections (themselves or by the partner) without any deleterious effect on stimulation cycles results and they expressed their will to continue for the next treatment cycles. CONCLUSION: After a quick initiation to subcutaneous injections, and its use, women appreciated their active involvement in the treatment and the self-sufficiency so achieved. By planning an educational program, medical staff could participate to improve the stress experienced by the women and their partners through the infertility treatment. PMID- 11732433 TI - [Methods for laparoscopy: open laparoscopy or closed laparoscopy? Attitude of the French Central University Hospital]. AB - Standard technique of insufflation of the pneumoperitoneum includes the use of the Veress needle followed by the blind insertion of the umbilical trocar. To avoid blind trocar insertion, numerous techniques for the creation of the pneumoperitoneum have been reported: open-laparoscopy and micro-laparoscopy are the two major alternative methods used in France. The aim of this study was to determine the incidence of open-laparoscopy in the French departments of Gynecology of the University Hospitals. With this purpose, we send to each chief of the department of Gynecology in a University Hospital a simple questionnaire about the methods used in his division for the creation of pneumoperitoneum. Sixty-three chiefs of department have answered to the questionnaire. Thirteen (21%) answered they only used conventional technique. Ten (16%) departments perfonned always open-laparoscopy, and 40 University Hospital mostly used conventional technique combined sometimes when the initial procedure of insufflation failed or seems to be too dangerous (Obesity, previous laparotomy...). Five surgeons used a lateral rather than umbilical insertion for the openlaparoscopy in case of previous laparatomy. Fifteen departments used micro-laparoscopy since 1995, and mostly since 1998. We conclude that no simple technique can claim to be overwhelmingly superior. It seems tous dangerous to impose or to condemn any methods. Keeping the choice of the technique, learning security and organizing the survey of our results remain our recommendations for the approaches of abdominal entry for laparoscopy. PMID- 11732432 TI - [Twin birth after preimplantation diagnosis for cystic fibrosis]. AB - OBJECTIVE: Cystic fibrosis is a common autosomal recessive disease most often caused by a deletion (delta F508) in the CFTR gene. It is the most common indication for preimplantaion genetic diagnosis which allows genetic analysis of embryos obtained after in vitro fertilization and transfer of unaffected embryos into the patient's uterus. PATIENTS AND METHODS: We report the first preimplantation genetic diagnosis performed in Strasbourg for a couple at risk of having a child affected by severe cystic fibrosis due to a homozygous delta F508 mutation. Three days after fertilisation, embryos obtained after intra cytoplasmic testiculare sperm injection were biopsied and analysed. PCR amplification of the genomic fragment containing the delta F508 locus allowed detection of the delta F508 mutation and transfer only of the unaffected embryos. RESULTS: Three embryos were transferred after this preimplantation genetic diagnosis. A twin pregnancy was obtained and the babies born from this cycle are both exempt from the mutation. CONCLUSIONS: Preimplantation genetic diagnosis for the cystic fibrosis delta F508 mutation is now available in our centre. In this report, we could resolve both the problem of infertility and the risk of transmission of a severe form of cystic fibrosis. Preimplantation genetic diagnosis is also available for other mutations involved in cystic fibrosis and also for other genetic diseases. PMID- 11732434 TI - [The ovary and insulin resistance]. AB - Insulin resistance appears to be responsible of approximately half of the cases of polycystic ovaries, the other half being probably provoked by an anomaly of the stimulation of ovaries by an excess of LH. Nevertheless, it is likely that in most cases the two factors conjugate. The excess of androgen production by the ovarian stroma is one of the major symptoms of this disease. Today, however, the diagnosis is carried mainly with the assistance of ultra-sounds which, besides the increased ovarian volume, have permitted to discover an increased ovarian stromal vascularity. Two essential datas derive from the whole works: the increased frequency of ovarian hyperstimulation syndrome and the great number of metabolic complications which requires an endocrinological supervision. But the most recent works focus on the extension to all ages of this form of pathology: from the intra-uterine life to the post menopause; and on the hereditary character of this disease. The mystery remains concerning the mechanism of the favourable effect in clomifene resistant PCOS, of surgical and laparoscopic methods of ovulation induction to which it may be useful to resort after mature consideration. More recently the benefit at the administration of metformine has been confirmed by several works. PMID- 11732435 TI - [Disseminated peritoneal leiomyomatosis: apropos of a case with immunohistochemical study]. AB - We report one case of leiomyomatosis peritonealis disseminata in a 40-year-old woman, G4P4, who never used oral contraceptives. She had relapsing uterine leiomyoma resected twice by hysteroscopy. Clinical findings and imaging studies concluded to peritoneal carcinomatosis. The patient had hysterectomy, salpingo oophorectomy, omentectomy and resection of an intestinal segment which surface is studied with firm nodules ranging in size from 1 to 50 mm. Microscopic examination showed a smooth-muscle cell proliferation with no mitosis, no atypia and no necrosis. Immunohistochemical analysis showed diffuse and strong labeling for progesterone receptors and weak labeling for estrogen receptors. The patient is well 16 months later. In the literature, over than 100 cases are reported, eight were malignant. PMID- 11732436 TI - [The binomial oncologist-psychoanalyst: the patient's journey]. AB - In oncological medicine, professionals work with patients in very separate areas of specialization. This specialization does not always seem the best approach, however, this kind of specialized care can benefit the patients. We would like, in the following text, to share with you a daily experience of an oncologist and a psychoanalyst treating a patient. First, we will present the oncologist's point of view: his way of understanding the patient's journey, his role which is sometimes difficult to accept and maintain between taking action and speaking, the richness and benefits of multidisciplinary care, the oncologist/patient relationship which unfolds over a long-term and finally, his stance facing what wee call the 'end of life'. Secondly, the psychoanalyst will consider her place in a medical service, working with women facing breast cancer. Through oncological case studies, wee will follow in detail the psychological work which unfolds during regular sessions between patients and the psychoanalyst. We will try to analyse the dynamic in the triad: oncologist/patient/psychoanalyst, and the potential journey from this triad to a triangulation, rich in potential for psychological elaboration and evolution. Both, the oncologist and the psychoanalyst will illustrate their work's pecularities and will try to show how their care and intervention in a case come together in one common goal: to be listening to both the physical and the psychological distress of their patients. PMID- 11732437 TI - [Local spermicidal contraception]. PMID- 11732438 TI - [Urodynamic exploration]. PMID- 11732439 TI - [Breech delivery or cesarean? Must breech delivery be condemned in the case of breech presentation at term?]. PMID- 11732440 TI - [Breech delivery or cesarean? Breech presentation at term: the arguments for cesarean]. PMID- 11732441 TI - [Standards, options and recommendations for the initial management of patients with malignant ovarian epithelial tumors (abridged version) ]. PMID- 11732442 TI - Generic substitution of anti-arrhythmic drugs. PMID- 11732443 TI - The PBR system. PMID- 11732444 TI - Aorto-iliac occlusive disease in the various population groups of South Africa. PMID- 11732445 TI - Morphine and chronic cancer pain. PMID- 11732446 TI - Racist editorial. PMID- 11732447 TI - Effects of crime on psychiatrists. PMID- 11732448 TI - Arousal from a semi-comatose state on zolpidem. PMID- 11732449 TI - Antiretroviral therapy and drug interactions. PMID- 11732450 TI - Leucocyte depletion of blood components. PMID- 11732451 TI - Meeting the challenge of health for all through public health education: a response from the University of the Western Cape. PMID- 11732453 TI - Running, the kidneys and drinking too much--the hyponatraemia of exercise. PMID- 11732452 TI - Mass immunisation campaigns in South Africa--the case for judicious timing and spacing. PMID- 11732454 TI - Shoe size, stature and caesarean delivery. PMID- 11732455 TI - Plant sterol/sterolin supplement use in a cohort of South African HIV-infected patients--effects on immunological and virological surrogate markers. PMID- 11732456 TI - Use of alternative medicine by patients with neurological disease. PMID- 11732457 TI - Peak rates of diuresis in healthy humans during oral fluid overload. AB - OBJECTIVE: To determine whether rates of intestinal fluid absorption and renal diuresis can match high rates of fluid ingestion in healthy humans exposed to oral fluid overload, thereby preventing the development of hyponatraemia either by reverse sodium movement across the intestine (the Priestley-Haldane effect) or by expansion of the extracellular fluid volume. METHODS: Changes in renal function and in plasma chemical measurements in response to an oral fluid overload (0.9-1.8 l/h x 3 h) were investigated in 6 healthy control subjects at rest, and in a subject with a history of exercise-induced symptomatic hyponatraemia, during both prolonged (160-minute) exercise and at rest. FINDINGS: All control subjects gained weight (2.7 +/- 0.2 kg, mean +/- standard error of mean (SEM)) because the rate of oral fluid intake exceeded the peak rate of urine production (778 +/- 39 ml/h). Blood volume rose by 7.1 (+/- 0.5)% and plasma sodium concentrations fell progressively from 144 +/- 2.6 to 136 +/- 1.1 mmol/l (P < 0.05) in the control subjects. Plasma potassium and angiotensin II concentrations were unchanged and creatinine clearance was normal (approximately 125 ml/min). Free water clearance reached a maximum of 11.2 +/- 0.9 ml/min after 2 hours. The increase in body mass could be accounted for by calculated or measured changes in extra- and intracellular fluid volumes. Similar changes were measured in the subject with a previous history of symptomatic hyponatraemia. CONCLUSION: The rate of intestinal fluid absorption appeared to match the rate of oral fluid ingestion and there was no evidence of fluid accumulation in the intestine with reverse sodium movement from the extracellular space into intestinal fluid. The results of this study are therefore at variance with the Priestley-Haldane hypothesis and suggest that reverse sodium movement did not contribute to the hyponatraemia induced by oral fluid overload in these subjects. Rather it appears that humans may have a limited capacity to excrete fluid at rates in excess of approximately 900 ml/h in response to higher rates of oral fluid intake. When the rate of intestinal fluid absorption matches the rate of fluid ingestion and exceeds the kidneys' maximum capacity for fluid excretion, the excess fluid accumulates in the extra- and intracellular fluid compartments, inducing the dilutional hyponatraemia of water intoxication. These findings may have relevance to other clinical conditions in which hyponatraemia develops in response to high rates of oral or intravenous fluid provision. PMID- 11732458 TI - Is prophylaxis of early-onset group B streptococcal disease appropriate for South Africa? AB - BACKGROUND: Early-onset group B streptococcal (GBS) disease in neonates can be prevented by the use of intrapartum chemoprophylaxis. There are two prevention strategies, one based on risk factors and the other on culture screening for GBS. This study sought to establish whether GBS chemoprophylaxis is appropriate in a developing country such as South Africa. METHODS: All neonates with early-onset GBS disease born at Johannesburg Hospital between 1 January 1995 and 21 December 1997 were reviewed. Data were collected prospectively between 1 January and 31 October 1998. Data included demographic information, obstetric information, disease characteristics, admission details and mortality. The approximate cost of implementing both strategies was determined. RESULTS: The overall incidence of early-onset GBS was 1.16 per 1,000 live births. The rate was significantly greater in 1998 compared with the previous years. Most of the babies were born preterm (70%), and 60% required admission to the neonatal intensive care unit (ICU) (a total of 81 ICU days). Twelve of the babies died. Assuming that chemoprophylaxis would reduce the number of ICU days by half, this would save an amount of R52,000. Culture-based chemoprophylaxis would cost R10 million, whereas an approach based on risk factors would cost R31,140. CONCLUSION: In conclusion, we feel that early-onset GBS disease is sufficiently prevalent in our unit to justify the implementation of a chemoprophylaxis strategy based on risk factors. Whether other units should adopt a similar approach would depend on the local incidence of early-onset GBS. PMID- 11732459 TI - Efficacy and cost-effectiveness of bronchial arterial embolisation in the treatment of major haemoptysis. AB - OBJECTIVE: To determine the efficacy and cost-effectiveness of bronchial artery embolisation (BAE) in the treatment of major and massive haemoptysis in HIV positive and negative patients with pulmonary inflammatory disease. METHODS: A retrospective review of patients admitted over a period of 24 months to Wentworth Hospital with major haemoptysis treated using BAE. RESULTS: Eighty-seven patients were treated (77 males, 10 females). Bilateral disease was present in 50 patients (57%). Thirty-two patients were HIV-positive (37%). Embolisation was successfully performed in 77 patients (88.5%), and failed for technical reasons in 10 patients (11.5%). There was only one procedural complication. Fifty-seven patients had a successful outcome, with cessation of haemoptysis within 24 hours (66.5%). Haemoptysis continued in 30 patients (34%) (20 patients embolised and the 10 patients who had failed procedures). Fourteen of these patients (16%) required lobectomy or pneumonectomy as an emergency procedure. Five patients (5.7%) died from respiratory failure or pulmonary haemorrhage. Twenty-four HIV-positive patients were successfully embolised. Costing of BAE, including a 2-day ICU and 3 day ward stay, was R6,720; together with surgical resection the cost was R14,170. CONCLUSIONS: BAE is an effective treatment for major and/or massive haemoptysis in patients with pulmonary inflammatory disease who are not surgical candidates. Patients who are HIV-positive are able to tolerate the procedure well. PMID- 11732460 TI - A new combined DTP-HBV-Hib vaccine--strategy for incorporation of Hib vaccination into childhood immunisation programmes. AB - OBJECTIVES: To evaluate the immunogenicity and reactogenicity of a pentavalent vaccine prepared by extemporaneously mixing diphtheria-tetanus pertussis hepatitis B vaccine (DTP-HBV) and lyophilised Haemophilus influenzae type B (Hib) tetanus conjugate vaccines in the same syringe, compared with the same vaccines given as separate, concomitant administrations. DESIGN: Open, randomised comparative study. SETTING: Durban, South Africa. SUBJECTS: A total of 120 healthy male and female infants were enrolled in the trial and randomised into two groups; group 1 received the combined administration (DTP-HBV-Hib), and group 2 received separate administrations of DTP-HBV and Hib vaccines. Vaccines were given as a three-dose primary vaccination course at 2, 4 and 6 months [corrected] of age. OUTCOME MEASURES: Antibody levels were measured using standard techniques and local and general solicited symptoms were recorded using diary cards. RESULTS: All subjects had seroprotective titres against diphtheria and tetanus; and antipolyribose-ribitol phosphate (PRP) titres > or = 0.15 microgram/ml 1 month after the final dose. A vaccine response (defined as post-vaccination titres > or = 15 ELISA (EL).U/ml in initially seronegative subjects; and as post vaccination titres > or = pre-vaccination titres in initially seropositive subjects) against the pertussis component was seen in 83% and 85% of subjects in the groups receiving combined and separate administration. No differences were seen in any of the geometric mean titres (GMTs) between the two administrations either 2 months after the second dose or 1 month after the final dose. There was no observed increase in reactogenicity in the group receiving the mixed administration. CONCLUSIONS: The results demonstrate that combined DTP-HBV-Hib vaccine is well tolerated and immunogenic. PMID- 11732461 TI - Distribution of blood lead levels in schoolchildren in selected Cape Peninsula suburbs subsequent to reductions in petrol lead. AB - OBJECTIVE: To determine blood lead levels among children attending schools in selected Cape Peninsula suburbs, and to assess the impact of a reduction in the lead content of petrol. DESIGN: A cross-sectional analytical study of children's blood lead levels and associated risk factors. SETTING: Selected inner city, suburban, and peri-urban schools in the Cape Peninsula, expected to have differing levels of environmental exposure to lead. SUBJECTS: Grade 1 schoolchildren for whom prior written parental consent had been obtained, and who were present at school on the day of the study. OUTCOME MEASURES: Blood lead levels (microgram/dl), associated with a wide range of potential risk factors. RESULTS: Median blood lead levels in suburbs varied from 14 to 16 micrograms/dl, the lowest levels occurring in the peri-urban suburb and the highest in the inner city suburb. Within the inner city suburb of Woodstock, variations in mean blood lead concentrations among schools were substantial, varying from 13 to 19 micrograms/dl. Overall, no change occurred in blood lead levels in this suburb subsequent to the lowering of the lead content of petrol. CONCLUSION: Every effort should be made in South Africa to control sources of lead in the urban environment. The study will serve as a useful baseline against which to measure the impact on blood lead levels of further actions which have been taken to promote the use of lead-free petrol in South Africa. PMID- 11732462 TI - 1998 survey of cardiac pacing in South Africa--report of the working group on registries of the cardiac arrhythmia society of South Africa (CASSA). AB - BACKGROUND: Permanent pacemakers provide effective relief of symptoms and are life-saving in patients with symptomatic heart block. Since pacemakers are only implanted by cardiologists or cardiothoracic surgeons in tertiary hospitals, the rates of pacemaker implantation provide a readily auditable measure of tertiary health care. METHODS: A survey was conducted of pacemaker implantation in South Africa in 1998, using questionnaires completed by implanters and information on the total number of pacemakers sold during the period, supplied by pacemaker distributors. RESULTS: A total of 1,643 new pacemakers were implanted in 1998 by 112 doctors working in 31 institutions (9 public and 22 private). The annual implant rate per million population was 39, compared with 31/million in 1995, an increase of 25.8%. Public hospitals accounted for 31.7% of primary pacemaker implants in 1998 as opposed to 37% in 1995. Practice in the public sector differed from that in the private sector in that atrioventricular (AV) block was the major indication for pacing in 75.3% versus 45.3%, whereas sinus node dysfunction accounted for 34.9% of private patients, as opposed to 16.2% of public hospital patients. Simple VVI single-chamber pacemakers accounted for 49.5% of public hospital implants versus only 9.6% in private patients. The reverse was true for dual-chamber implants (12.1% v. 42.3%). Severe cost constraints in the public sector have resulted in increasing use of cheaper single-chamber pacemakers for more stringent indications (predominantly AV block). The overall implant rate remains low compared with a median of 283/million in Europe. Large discrepancies persist between race groups (232/million whites, as opposed to 8.8/million blacks) and regions (89.3/million in the Western Cape versus 10.8/million in the four provinces without pacemaker implanters). CONCLUSION: It is likely that socio-economic factors play a major role in the unequal distribution of this highly effective treatment for potentially lethal bradyarrhythmias. The changing trends in pacemaker implantations indicate a shift of tertiary health care resources from the public to the private sectors. PMID- 11732463 TI - [High or middle dose tinzaparin versus aspirin for the treatment of cerebral ischemic accident]. PMID- 11732464 TI - [Cognitive restraint in obesity. History of ideas, clinical description]. AB - AN EXPLANATION OF DISAPPOINTING RESULTS: Handling obesity and moderate overweight with dietary prescriptions or cognitive and behavioral therapies gives unsatisfying results. For some, such treatments even have the drawback of aggravating eating behavioral patterns, not to mention other psychopathological disorders. We believe, on the contrary, that such disappointing results and troubles may be explained by the cognitive restraint theory. COGNITIVE RESTRAINT THEORY: After the radical questioning of a psycho-genic origin to obesity, the efforts made to lose weight have gradually been held responsible for the psychopathological troubles observed in the cases of overweight patients. Herman and Polivy thus consider that slimming diets bring about a state of cognitive restraint, i.e. a way of eating ruled by beliefs of all kinds in the various consequences on the patients' weight of alimentary habits and choices, as opposed to inner criteria of hunger and satiety. DIFFERENT STATES: A person in a state of cognitive restraint can either experience inhibition without loss of control, or a loss both of inhibition and of control. In the former case, one can distinguish: a) a voluntarist stage in which the individual deliberately chooses not to heed his hunger and satiety sensations in order to privilege rules that are supposed to allow him/her to control his/her weight; b) an unconscious stage during which physiological sensations are blurred, and eating habits ruled by unconscious cognitive processes and emotions. The individual thus ends up organising his eating behaviour around his/her fear of lacking, the frustration/guilt doublet and troubles in the comforting pattern. Such a state of inhibition is frequently interspersed with losses of control, described as hyperphagic or bulimic bouts and compulsive eating. THERAPEUTIC STRATEGIES: Certain authors, facing the necessity to bring down their patients' weight, tend nevertheless to advocate restrictive methods, or seem to consider that in spite of the above-mentioned drawbacks, cognitive restraint remains the only extant method to lose weight. An alternative therapeutic strategy is definitely worth exploring: one that consists precisely in helping the overweight patient to deliver him/herself from cognitive restraint. The first target could thus be to restore eating habits in conformity with personal tastes and with the patient's own personal hunger and satiety physiological regulation systems. This implies a questioning of dysfunctioning cognitive processes, and a thorough investigation of upholding factors such as self-esteem or emotional and relational troubles. PMID- 11732466 TI - [Pneumothorax in distomatosis]. AB - INTRODUCTION: Distomatosis is a parasite infection following ingestion of food contaminated by Fasciola hepatica, the tropism of which is essentially hepatic. The mechanism of this exceptional complication is discussed in two cases of pneumothorax. OBSERVATIONS: Two 35 and 60 year-old patients presenting spontaneous pneumothorax during distomatosis, the first revealing the disease, the second suggesting the evolution of a severe form of the disease. The surgical biopsy in the first patient revealed a granulomatous pleuro-pneumopathy rich in giant cells. DISCUSSION: Pulmonary manifestations of distomatosis are frequent during the invasive phase. Their immunoallergic origin can be questioned and emphasis should be placed on lesions secondary to aberrant parasite migrations. PMID- 11732465 TI - [Predictive factors of response to buprenorphine in the substitutive treatment of heroin addicts. Results of a multicenter study of 73 patients]. AB - OBJECTIVE: Buprenorphine has a partial morphine-agonist pharmacological profile. It is proposed as alternative to methadone in opiate drug addicts with greater safety of use and less cost in terms of public health. The aim of this study was to determine the clinical factors of response to this molecule. METHOD: The study was conducted in 73 patients treated for 3 months with adaptable doses. Mean dose was 8.5 mg/d (range: 3 to 16 mg/d). Response to treatment was defined as: still in the study at 3 months and absence of opiates in 75% of urinary samples over the past month. RESULTS: Forty-eight patients responded and 25 did not. The determinating clinical variables of response were: opiate drug addiction less than 10 years, high score on the Addiction Severity Index (ASI), absence of depression according to the Minnesota Multiphasic Personality Inventory (MMPI) and low rate of disinhibition on Zukerman's sensation seeking scale. Conversely, the dose of buprenorphine within the limits specified in the Marketing Authorisation did not intervene in the response. CONCLUSION: In view of its partial agonist effect, administration of buprenorphine must be reserved for patients addicted to opiates for less than 10 years, non-depressive and with low disinhibition on Zukerman's scale. PMID- 11732467 TI - [Purpura fulminans and coxsackie B2 co-infection]. AB - INTRODUCTION: During septic shock in children, myocardial deficiency usually lasts 3 to 6 days. When prolonged, an other etiology should be looked for. OBSERVATION: A 29 month-old child presented with purpura fulminans, probably due to Neisseria meningitidis. Positivity of the serodiagnostic for coxsackie B2, performed because of persisting myocardial deficiency, evoked diagnosis of a coxsackie B2 co-infection. DISCUSSION: Abnormally prolonged myocardial deficiency during purpura fulminans must lead to the search for a congenital, metabolic or infectious cardiomyopathy. PMID- 11732468 TI - [Recommendations of "scholar societies" and their scientific bases]. PMID- 11732469 TI - [Anti-pneumococcal vaccination in splenectomized patients. Evaluation of its use from 1995 to 2000 at the CHU of Nice]. PMID- 11732470 TI - [Malaria in Haiti, an ever-present endemic disease]. PMID- 11732471 TI - [Symptoms notified by people living near cell phone relay stations]. PMID- 11732472 TI - [Ovarian hyperstimulation syndrome with cerebral arterial thrombosis treated with fibrinolysis]. PMID- 11732473 TI - [Risks associated with exposure to cytostatics in health care personnel]. AB - RISKS RELATED TO HANDLING: Cytostatic drugs (CS) destroy malignant cells. However, they also have deleterious effects on healthy cells. Manufacturing staff and nurses may suffer from side effects when handling these molecules. WHAT TYPE OF RISKS?: Laboratory staff and nurses handling these molecules are exposed to sub-therapeutic concentrations of these products, which induce little known biological effects. Cytostatics can provoke allergies, mutations and probably have carcinogenic and teratogenic effects. ASSESSMENT OF EXPOSURE: Exposure can be assessed by calculating the index of cytostatic contact. Medical surveillance includes measurement of he surrounding metrology and specific biometry. PREVENTION: Basic preventive measures include the wearing of masks, gloves and overalls and the installation of vertical laminar flow hoods. PMID- 11732474 TI - [Human parasitoses in French Guiana]. AB - MALARIA: This review summarises current knowledge of the epidemiological situation regarding parasitoses in French Guiana. Malaria is the most prevalent infection. The most common species is Plasmodium falciparum which is responsible for three quarters of all bouts of malaria. Plasmodium vivax is mostly present in endemic areas inhabited by Amerindian communities. Plasmodium malariae is rarer. CHAGAS' DISEASE, LEISHMAMIASIS AND TOXOPLASMOSIS: Four acute cases of cardiac Chagas' disease were observed between 1994 and 1996. These cases serve as a warning concerning the possible emergence of this type of parasitosis among the poor in Latin America. Cutaneous leishmaniasis is still highly endemic, with an incidence of about 2 cases per 1,000 inhabitants, which has remained essentially unchanged for 15 years. The identification of severe primary toxoplasmosis in immunocompetent individuals in recent years seems to be associated with an as yet poorly known, natural, Amazonian cycle. INTESTINAL PARASITOSES: Improvements in living conditions, hygiene and treatments are such that intestinal parasitoses, other than anguilluliasis, are no longer a major problem in the urban or coastal areas. The situation is very different in the central areas of the country and in the Amerindian and black half-cast communities, with high prevalences of ankylostomiasis (hookworm disease), anguilluliasis and amoebal disease (Entamoeba histolytica/E. dispar). OTHERS: Lymphatic filariasis is no longer endemic; tropical schistosomiasis and distomiasis have never been endemic. Parasitic pests (cutaneous larva migrans and insect or dust-mite ectoparasites) are more spectacular than severe, and constitute a classic "exotic" component of the spectrum of diseases in French Guiana. PMID- 11732475 TI - Negotiating the maze: problem based, collaborative distance learning in dentistry. PMID- 11732476 TI - Preface. Certain arts have long been associated with attendant diseases. PMID- 11732477 TI - Grow and go: make sure career-building moves help you and your practice. PMID- 11732478 TI - Can patients get through your practice's electronic 'front door'? PMID- 11732479 TI - Incentive programs that drive your employees to excel. PMID- 11732480 TI - Bad charts: the malpractice plaintiff attorney's best friend. PMID- 11732481 TI - Public relations on a shoestring budget. PMID- 11732482 TI - Want better returns, lower costs? Start tracking key data today. PMID- 11732483 TI - Defining the profession. "Just what do you think you're doing?". PMID- 11732484 TI - ACOG Practice Bulletin. The use of hormonal contraception in women with coexisting medical conditions. Number 18, July 2000. AB - Although numerous studies have addressed the safety and effectiveness of hormonal contraceptive use in healthy women, data are far less complete for women with underlying medical problems or other special circumstances. Because recommendations vary widely, substantial confusion exists with respect to contraceptive guidelines for women with coexisting medical conditions or other concerns. Using available scientific evidence, this Practice Bulletin will provide information to facilitate contraceptive counseling and selection for women with coexisting medical conditions. PMID- 11732485 TI - Anderson-Fabry disease: clinical manifestations and impact of disease in a cohort of 60 obligate carrier females. PMID- 11732486 TI - Thrombophilic polymorphisms in pre-eclampsia: altered frequency of the functional 98C>T polymorphism of glycoprotein IIIa. PMID- 11732488 TI - Maternal gene effect in neurofibromatosis 2: fact or artefact? PMID- 11732487 TI - Cystic fibrosis patients with the 3272-26A>G splicing mutation have milder disease than F508del homozygotes: a large European study. PMID- 11732489 TI - Familial medullary thyroid carcinoma and prominent corneal nerves associated with the germline V804M and V778I mutations on the same allele of RET. PMID- 11732490 TI - Comprehension of cancer risk one and 12 months after predictive genetic testing for hereditary non-polyposis colorectal cancer. PMID- 11732491 TI - Population prevalence and estimated birth incidence and mortality rate for people with Prader-Willi syndrome in one UK Health Region. PMID- 11732492 TI - Mosaicism for 45,X cell line may accentuate the severity of spermatogenic defects in men with AZFc deletion. PMID- 11732493 TI - Alternative centromeric inactivation in a pseudodicentric t(Y;13)(q12;p11.2) translocation chromosome associated with extreme oligozoospermia. PMID- 11732495 TI - Eliminating discrimination. PMID- 11732494 TI - Combating the labor shortage. PMID- 11732496 TI - Facilities upgrades improve bottom line. PMID- 11732497 TI - Recruiting online. PMID- 11732498 TI - Elements of a successful job search. PMID- 11732499 TI - Take the initiative to manage your career. PMID- 11732500 TI - Wait watcher. Lies, damned lies and spinners. PMID- 11732501 TI - [Distribution of Maori health services]. PMID- 11732502 TI - Bibliography of secondary sources on the history of dermatology. III. Books, monographs, and chapters in English supplemented through 2000. PMID- 11732503 TI - Inner visions. Waiting to exhale. PMID- 11732504 TI - Fetal exposure to tobacco smoke is common. PMID- 11732505 TI - Cross-contamination: HS-Sultan is not a myeloma but a Burkitt lymphoma cell line. PMID- 11732506 TI - Traumatic lumbar puncture at diagnosis and outcome in childhood acute lymphoblastic leukemia. PMID- 11732507 TI - Transcobalamin polymorphism and homocysteine. PMID- 11732508 TI - Dependence of age-specific incidence of acute myeloid leukemia on karyotype. PMID- 11732509 TI - Total eversion of the upper eyelids in a newborn. AB - A case of bilateral congenital eversion of the lids in a newborn is presented. The baby presented at the Eye Clinic of Otibhor Okhae Teaching Hospital about four hours after a spontaneous vaginal delivery. Surgical correction was carried out one eye at a time with absorbable sutures. Recovery was uneventful and normal anatomical configuration was regained 8-10 days post operatively. The aim of this case report is to create awareness among workers in maternity homes, primary health centres and other health institutions about the existence of this congenital anomaly. Prompt referral to the Ophthalmologist is advised, as the condition is very amenable to treatment. PMID- 11732510 TI - Conflicts of interest: are they a problem for anaesthesia journals? What should we do about them? PMID- 11732511 TI - Skin testing and the anaesthetist. PMID- 11732512 TI - Model-based automatic feedback control versus human control of end-tidal isoflurane concentration using low-flow anaesthesia. AB - We studied the clinical use of an automatic feedback control system to adjust the end-tidal anaesthetic concentration with a low-flow method. The end-tidal controller uses two input signals (the end-tidal and inspiratory concentrations) to control the isoflurane concentration in the fresh gas flow, using a model based algorithm. We studied 22 ASA I-III patients during elective surgery lasting more than 2 h. The anaesthetist was asked to make four step changes of the target end-tidal concentration (+0.3, +0.6, -0.3, -0.6 vol%), either manually (Group A) or by setting the target value for the feedback controller (Group B), and then the control was changed and the step changes were repeated, in a crossover design. Eighty step changes with each control method were compared in terms of response time, maximal overshoot and stability. The automatic control system was more accurate and stable than the human controller for step increases and step decreases, with less overshoot/undershoot and greater stability [e.g. maximal overshoot 14.7 (SD 3.7)% and 18 (8.1)% respectively for +0.6 vol% step changes, and 19.8 (3.7)% and 30.7 (13.2)% respectively for +0.3 vol% step changes]. However, the automatic control system showed a faster response time than the manual method only with large increasing steps (e.g. 149 (32) s and 205 (57) s respectively for +0.6 vol% step changes) and was not different from manual control for decreasing steps. Automatic control of the end-tidal isoflurane concentration can be better than human control in a clinical setting, and this task could be done automatically. PMID- 11732513 TI - Ropivacaine 0.2% versus bupivacaine 0.1% with fentanyl: a double blind comparison for analgesia during labour. AB - We have performed a randomized, double-blind comparison of two epidural drug regimens for analgesia in labour. In the bupivacaine group (BUPIV), 101 healthy parturients received 0.1% bupivacaine with fentanyl 2 microg ml(-1). In the ropivacaine group (ROPIV), 102 women received 0.2% ropivacaine. Both groups received an initial loading dose of 15 ml, a continuous infusion of 8 ml h(-1), and top-ups of 10 ml. Breakthrough pain not responding to a routine top-up was treated with an 'escape' top-up of 10 ml 0.25% bupivacaine. The two groups were compared for complete analgesia at 30 min, routine and 'escape' top-up requirements, midwife assessment of analgesic efficacy, delivery mode, patient visual analogue scores (VAS) for first and second stage analgesia, overall satisfaction, and patient assessment of motor blockade. Patients receiving ropivacaine received fewer routine top-ups (median 1.0 vs. 2.0, P=0.001) and fewer escape top-ups (9.8% vs. 21.8%, P=0.02). The ropivacaine group was more likely to be pain free in the first stage (51% vs. 33.7%, P=0.01). There were no significant differences in patients' assessment of motor block or mode of delivery between the groups. Pain relief and satisfaction scores from midwives and patients were consistently better in the ropivacaine group, but did not reach statistical significance. PMID- 11732514 TI - Ropivacaine plasma concentrations during 120-hour epidural infusion. AB - The pharmacokinetics of ropivacaine were evaluated during long-term continuous epidural analgesia (CEDA) for about 120 h. The total and free plasma concentrations of ropivacaine and the alpha1-acid glycoprotein (AAG) concentration were measured in 12 patients after total knee arthroplasty. The infusion rate was adjusted according to patients' analgesic needs or side effects. The mean (SD) rate of infusion of ropivacaine (Naropin 2 mg ml(-1)) was 14.6 (3.2) mg h(-1) on the day of surgery and was increased after surgery to 15.4 (4.4) mg h(-1) on days 1-5. This was equivalent to an absolute dose of 1786 (553) mg of ropivacaine over the entire infusion period. After an initial increase, the mean free ropivacaine plasma concentration nearly plateaued and than decreased slightly after approximately 70 h. The individual peak free plasma concentration was 0.096 (0.034) microg ml(-1). The highest individual free plasma concentration was 0.16 microg ml(-1). The individual peak total plasma concentration, 4.1 (1.2) microg ml(-1), was achieved after 67.7 (16.5) h, although the AAG concentration increased throughout the observation period. Our data support the safety and efficacy of long-term ropivacaine CEDA. PMID- 11732515 TI - Pain management after adenoidectomy with ketoprofen: comparison of rectal and intravenous routes. AB - We compared the efficacy of rectally and intravenously administered ketoprofen for pain management after day-case adenoidectomy. Patients (123 children aged 1-9 yr) were allocated randomly to receive on induction of anaesthesia ketoprofen 25 mg rectally with an i.v. placebo, ketoprofen 25 mg i.v. with a rectal placebo, or placebo both i.v. and rectally. The method of anaesthesia and the operative technique were standardized. Postoperative pain was assessed at rest and during swallowing using the Maunuksela pain scale (0=no pain, 10=worst possible pain). Fentanyl 0.5 microg kg(-1) was given as rescue analgesia. There was no significant difference between the two ketoprofen groups in their requirement for rescue analgesics. However, both the proportion of children needing rescue analgesics [55 of 84 children (65%) vs. 33 of 39 children (84%); difference 19%, 95% confidence interval 4-34%, P=0.029] and the number of rescue analgesic doses [mean 1.2 (SD 1.2) vs. 2.2 (1.4); mean difference 0.9, 95% confidence interval 0.4-1.4, P=0.001] were significantly lower among children receiving ketoprofen than in children receiving placebo. Adverse events, duration of operation, perioperative bleeding, pain scores and time of discharge were similar in the three groups. PMID- 11732516 TI - Sonographic measurement of needle insertion depth in paravertebral blocks in women. AB - Single-injection paravertebral block offers adequate unilateral analgesia for thoracic and upper abdominal surgery. This technique is easy to learn but there is a risk, albeit low, of pleural puncture. The aim of the study was to determine whether sonographic measurements of the distances from the skin to the transverse process and to the parietal pleura are useful for calculating the required depth of needle insertion. Before puncture of the paravertebral space, the distances from the skin to the transverse process and to the parietal pleura were measured by sonography. The deviation of the needle from the horizontal plane was measured and an angle correction for the insertion depth was calculated. Twenty-two women undergoing elective unilateral breast surgery were studied. Sonographic visualization of the transverse process and the parietal pleura and measurement of their distances from the skin was successful in all women. Puncture of the paravertebral space failed in one obese woman. There was a very close correlation between needle insertion depth from the skin to the transverse process and the distance measured by ultrasound if angle correction was used (adjusted r2=0.95). Similarly, there was excellent correlation between the angle-corrected ultrasound distance from the skin to the parietal pleura and the distance from the skin to the paravertebral space (adjusted r2=0.92). PMID- 11732517 TI - Weal and flare responses to intradermal rocuronium and cisatracurium in humans. AB - Thirty volunteers underwent intradermal skin testing with increasing concentrations of rocuronium and cisatracurium to evaluate weal and flare responses, and whether either agent would cause mast cell degranulation and sensitization upon re-exposure. We found that intradermal injection of rocuronium and cisatracurium at concentrations > 10(-4) M resulted in positive weal (>8 mm) responses, and positive flare responses at > 10(-4) and > 10(-5) M respectively. Only cisatracurium caused mild to moderate mast cell degranulation, and neither drug caused significant in vitro histamine release from whole blood collected from study subjects 4 weeks after skin testing. Skin testing with rocuronium and cisatracurium should be performed at concentrations < 10(-4) and < 10(-5) M respectively to avoid false-positive responses. The ability of these agents to produce positive weal and flare responses at relatively low concentrations may explain the high incidence of potential reactions reported. PMID- 11732518 TI - Pharmacokinetics and dynamics of atracurium infusions after paediatric orthotopic liver transplantation. AB - We examined the pharmacokinetics and pharmacodynamics of atracurium besylate and its metabolites in children after orthotopic liver transplantation (OLT), as a suitable model for critically ill children. Ten children were studied after OLT on return to the intensive care unit (ICU). The mean (range) age was 36 (7-78) months, and weight 6-24.2 kg. Atracurium was started at induction of anaesthesia and adjusted in the ICU according to clinical need. Neuromuscular block was measured using accelerometry (TOFguard) and the train-of-four (TOF) ratio or count. Arterial plasma samples for atracurium and metabolites taken before, 12 hourly during, and at frequent intervals after the infusion were analysed by HPLC. The mean (range) maximum infusion rate during steady-state conditions was 1.44 (0.48-3.13) mg kg(-1) h(-1) and the duration of infusion 36.9 (22.5-98.4) h. Tachyphylaxis was not observed. The mean terminal half-life (t1/2) for atracurium was 18.8 (12-32.3) min. The steady-state plasma clearance (CLss) was 13.9 (7.9 20.3) ml min(-1) kg(-1) and the terminal volume of distribution (Vz) 390 (124 551) ml kg(-1); both were higher than in adults after successful OLT. The maximum concentration (Cmax) of laudanosine was 1190 (400-1890) ng ml(-1) and t1/2 was 3.9 (1.1-6.7) h. The renal clearance of laudanosine was 0.9 (0.1-2.5) ml min(-1) kg(-1) and increased with urine flow, but there was no significant relationship with serum creatinine. EEG spikes were confirmed in one child only; the corresponding laudanosine Cmax was 720 ng ml(-1). Monoquaternary alcohol Cmax was 986 (330-1770) ng ml(-1) and t1/2 42.9 (30-57.7) min. Mean recovery time on stopping the atracurium infusion to a TOF ratio >0.75 was 23.6 (12-27) min. Atracurium is an effective and safe neuromuscular blocking agent in this population. Laudanosine concentrations are not excessive if graft function is satisfactory. PMID- 11732519 TI - Simultaneous determination of neuromuscular block at the larynx, diaphragm, adductor pollicis, orbicularis oculi and corrugator supercilii muscles. AB - We simultaneously determined the neuromuscular blocking effect of mivacurium 0.2 mg kg(-1) at five muscles in 20 women undergoing gynaecological surgery. Evoked electromyographic responses were obtained using surface electromyography (EMG) at the adducting laryngeal muscles, the diaphragm (lateral to vertebrae T12/L1 or L1/L2) and the adductor pollicis muscle and acceleromyographic (AMG) responses were measured at the orbicularis oculi and the corrugator supercilii muscle. Onset time and times for the first twitch response (T1/T0) to return to 25, 75 and 90% at the adducting laryngeal muscles and the diaphragm were significantly (P<0.005) shorter than at the adductor pollicis, the corrugator supercilii or the orbicularis oculi muscles (mean (SD) onset time: 89 (26) s and 78 (17) s to 202 (45) s, 152 (41) s, 194 (40) s; T1/T0=25%: 10.4 (1.5) and 11.4 (1.2) min versus 20.5 (3.9), 15.9 (3.3), 16.3 (3.7) min; T1/T0=90%: 15.5 (1.6) and 16.1 (1.6) min versus 27.4 (4.6), 21.5 (3.8), 23.3 (5.1) min). Onsetand clinical duration of neuromuscular block at the larynx and the diaphragm after mivacurium 0.2 mg kg( 1) are shorter than in the peripheral muscles. Monitoring of neuromuscular block in the diaphragm was successfully used in all patients. PMID- 11732520 TI - Pre-operative oral erythromycin reduces residual gastric volume and acidity. AB - We investigated whether low-dose erythromycin (200 mg) given as an oral premedicant altered the residual gastric volume and its acidity in fasted patients at induction of anaesthesia in a single-blinded study. Sixty patients were allocated randomly to receive either an erythromycin tablet (200 mg) or nothing with 10 ml water 3 h before induction of anaesthesia, and another 60 patients 1 h before induction of anaesthesia. Oral erythromycin significantly reduced residual gastric volume when it was given approximately 3 h (P<0.05; 95% CI for median difference: 0.1-17 ml) or 1 h (P<0.0005; 95% CI for median difference: 6-24 ml) before induction of anaesthesia. Erythromycin significantly reduced gastric acidity when it was given 1 h before induction of anaesthesia (P<0.02; 95% CI for median pH difference: 0.1-1.7). In contrast, when given 3 h before induction of anaesthesia, erythromycin did not significantly alter acidity. PMID- 11732521 TI - Dexamethasone for prophylaxis of nausea and vomiting after epidural morphine for post-Caesarean section analgesia: comparison of droperidol and saline. AB - We have evaluated the prophylactic effect of i.v. dexamethasone 8 mg in preventing nausea and vomiting during epidural morphine for post-Caesarean section analgesia. Droperidol 1.25 mg and saline served as the control. We studied 120 parturients (n=40 in each group) receiving epidural morphine for post Caesarean section analgesia, in a randomized, double-blind, placebo-controlled study. All parturients received epidural morphine 3 mg. Both dexamethasone and droperidol significantly decreased the total incidence of nausea and vomiting compared with saline, with incidences of 18, 21 and 51% for the three treatments respectively (P<0.01 and P<0.05 respectively). Parturients who received droperidol reported a more frequent incidence of restlessness (16%) than those who received dexamethasone (P<0.05). PMID- 11732522 TI - Central cholinergic depletion induced by 192 IgG-saporin alleviates the sedative effects of propofol in rats. AB - We examined the effect of central cholinergic depletion on the sedative potency of propofol in rats. Depletion was produced by intracerebroventricular administration of an immunotoxin specific to cholinergic neurones (192 IgG Saporin; 2 microg). As a result of this lesion, acetylcholine concentration was reduced by about 40% in the frontoparietal cortex and in the hippocampus but was essentially normal in the striatum and cerebellum. Sedation in rats was assessed as the decrease in locomotor activity. Sedative potency of propofol (30 mg kg(-1) i.p.) was reduced by about 50% in rats who received the injection of 192 IgG Saporin as compared to controls. These results show that a central cholinergic depletion alleviates the sedative effect of propofol, and indicates that basal forebrain cholinergic neurones might mediate part of the sedative/hypnotic effects of propofol. PMID- 11732523 TI - Inhibitory effects of intravenous anaesthetic agents on K+-evoked norepinephrine and dopamine release from rat striatal slices: possible involvement of P/Q-type voltage-sensitive Ca2+ channels. AB - The role of the voltage-sensitive Ca2+ channel (VSCC) as a target for anaesthetic action remains controversial. In this study we characterized the VSCC subtypes involved in K+-evoked norepinephrine and dopamine release from rat striatal slices and used this model system to examine the effects of a range of i.v. anaesthetics on release. Nifedipine (L-channel-selective), omega-conotoxin GVI(A) (N-channel-selective), omega-agatoxin IV(A) (P-channel-selective), omega conotoxin MVIIc (P/Q-channel-selective) and Cd2+ (non-selective), along with alphaxalone, propofol and ketamine, were used in various combinations. Omega Agatoxin IV(A), omega-conotoxin MVIIc and Cd2+ fully (100%) inhibited norepinephrine and dopamine release. Clinically achievable concentrations of alphaxalone inhibited norepinephrine and dopamine release, with concentrations producing 25 and 50% inhibition (IC25 and IC50) of the maximum of 2.1 and 7.8 microM respectively for norepinephrine and 2.9 and 7.2 microM for dopamine. The effects of propofol were observed at the top of the clinical range and those of ketamine exceeded this range. In addition, IC50 values for alphaxalone in the presence and absence of nifedipine and omega-conotoxin GVI(A) did not differ from the control. Our data suggest that clinically achievable concentrations of alphaxalone and propofol inhibit norepinephrine and dopamine release, which is mediated predominantly through P/Q-type VSCCs, suggesting a role for these channels in anaesthetic action. PMID- 11732524 TI - Spinal and supraspinal midazolam potentiates antinociceptive effects of isoflurane. AB - The effects of lumbar intrathecal (i.t.) and intracerebroventricular (i.c.v.) midazolam on nociception during isoflurane anaesthesia were studied in rats using the tail-flick test. Rats received i.t. midazolam 2 and 4 microg or i.c.v. midazolam 4 and 8 microg during 1.1, 1.2 and 1.3% isoflurane or without isoflurane. Neither i.t. nor i.c.v. midazolam alone at doses studied influenced nociceptive responses. 1.1% isoflurane showed a minimum antinociceptive effect which was not influenced by i.t. or i.c.v. midazolam. 1.2 and 1.3% isoflurane produced moderate antinociception which was markedly potentiated by both i.t. and i.c.v. midazolam. The effects of midazolam shown in the present study are different from the reported effects of midazolam on opioid-induced antinociception; where spinally administered midazolam potentiates and supraspinal midazolam inhibits the antinociceptive effects of morphine. The present results suggest that midazolam potentiates isoflurane-induced antinociception at doses where no effect is seen alone. PMID- 11732525 TI - Production of human albumin solution: a continually developing colloid. PMID- 11732526 TI - Increased pre-operative platelet counts are a possible predictor for reduced sensitivity to heparin. AB - We investigated a possible relationship between pre-operative platelet count and reduced sensitivity to heparin in 87 patients undergoing cardiac surgery with cardiopulmonary bypass (CPB). Sensitivity to heparin was determined by measuring the slope of the heparin dose response (HDR) before surgery. Pre-operative platelet counts were measured as part of routine analysis of the patients' coagulation status. Patients with an HDR slope of <80 s u. ml(-1), were considered to have a reduced sensitivity to heparin and activated clotting time data were collected from these patients before and after heparin administration before CPB. A significant correlation was determined between pre-operative platelet levels and HDR slope (P<0.001). Platelet counts were significantly greater in heparin-resistant patients compared with those who had the expected response to the anticoagulant (P<0.05). This could be caused by an increased capacity to produce platelet factor 4, which neutralizes heparin. PMID- 11732527 TI - Prevention of postoperative nausea and vomiting by continuous infusion of subhypnotic propofol in female patients receiving intravenous patient-controlled analgesia. AB - In this prospective, randomized, double-blind, placebo-controlled study, the use of continuous subhypnotic propofol infusion as an antiemetic in fentanyl intravenous patient-controlled analgesia (i.v. PCA) was investigated during the first 24 h after surgery. One hundred female patients, ASA I-II, aged 20-71 yr, undergoing major gynaecological or orthopaedic surgery, were included. Either propofol 10 mg or placebo (1 ml of Intralipid) was given and one of the following five regimens was maintained for 24 h: propofol 5, 10, 15 or 20 microg kg(-1) min(-1) or Intralipid 1 ml h(-1) as a placebo. Fentanyl i.v. PCA was started in the postanaesthesia care unit for postoperative analgesia. Significantly more of the patients given propofol 15 and 20 microg kg(-1) min(-1) experienced no nausea or vomiting compared with those given placebo (65% and 70% versus 25%; P<0.05). Patients given propofol 20 microg kg(-1) min(-1) reported more sedation than those in the other groups 4 h after surgery (P<0.05). PMID- 11732528 TI - Intravenous anaesthesia and the rat microcirculation: the dorsal microcirculatory chamber. AB - The use of the dorsal microcirculatory chamber in male Wistar rats (n=7) to study the effects of induction and maintenance of anaesthesia on the microcirculation is described. Different patterns of responses were observed. At induction, arteriolar dilation was found following propofol and thiopental but ketamine produced constriction. During maintenance, constriction of arterioles was seen with ketamine and thiopental but dilation persisted with propofol. The dorsal microcirculatory chamber appears to be a useful tool for the study of microcirculatory changes related to anaesthesia. PMID- 11732529 TI - Serotonergic receptor antagonists alter responses to general anaesthetics in rats. AB - Serotonergic neurotransmission is involved in controlling arousal levels in humans and other animals. Here, the effects of serotonergic receptor antagonists on the induction and depth of anaesthesia produced by three different general anaesthetics were investigated. Rats were pretreated (i.p.) with either methiothepin (1.5 mg kg(-1)), mianserin (5 mg kg(-1)), ketanserin (7 mg kg(-1)) or saline. Subsequently, successive, cumulative doses (i.p.) of either ketamine (final, cumulative dose of 350 mg kg(-1)), sodium pentobarbital (final dose 77 mg kg(-1)), or chloral hydrate (final dose 600 mg kg(-1)) were administered. The response to the anaesthetics was measured using a behavioural test battery assessing nocifensive reflexes and hypnotic state. Pre-treatment with methiothepin enhanced responses to all three anaesthetics; mianserin enhanced responses to chloral hydrate. These results show that some serotonergic receptor antagonists change anaesthetic requirements, resulting in enhanced anaesthesia to hypnotics with different mechanisms of action. PMID- 11732530 TI - Sex and oestrous cycle differences in visceromotor responses and vasopressin release in response to colonic distension in male and female rats anaesthetized with halothane. AB - Visceromotor responses and vasopressin release before and after colonic visceral distension were compared between male (n=5 (n=4 for vasopressin)) and female rats and between females during the oestrous cycle (proestrus n=6, oestrus n=5, metestrus n=5, diestrus n=6) at a controlled depth of anaesthesia. Pre stimulation vasopressin and blood pressures demonstrated oestrous cycle variability. The mean (SEM) colonic balloon pressure triggering visceromotor responses was significantly higher in males (64 (4) mm Hg) than females (41 (1) mm Hg), P=0.002 and within females, proestrus rats had the lowest thresholds, (29 (1) mm Hg, P<0.01). Post-stimulation, vasopressin concentrations increased significantly in all groups (males 1.34 (0.39) to 2.24 (0.74) pmol litre(-1); females 1.54 (0.24) to 2.88 (0.58) pmol litre(-1); P=0.002). Within groups statistically significant differences were measured in proestrus 2.06 (0.56) to 3.42 (1.12) and oestrus 1.16 (0.38) to 2.76 (0.60) pmol litre(-1) (P<0.05). High vasopressin concentrations coupled with low-pressure stimulation during proestrus shows sex-hormone dependent integration of the neuroendocrine response to noxious visceral stimulation. PMID- 11732531 TI - Survivors of childhood cancers: implications for obstetric anaesthesia. AB - Treatment of many childhood malignancies involves surgery, radiotherapy and chemotherapy. If the child survives, normal physical development can be impaired and abnormalities with anaesthetic implications may be present. We discuss two women with a range of problems who presented for obstetric anaesthesia, having survived childhood malignancies. Common features included anthracycline cardiotoxicity and short stature. Both patients received incremental spinal anaesthesia in order to titrate the dose of local anaesthetic required to produce an adequate block height and to minimize cardiovascular instability. PMID- 11732532 TI - Prolonged vecuronium neuromuscular blockade associated with Charcot Marie Tooth neuropathy. AB - Charcot Marie Tooth (CMT) disease comprises a group of disorders characterized by progressive distal muscle weakness and wasting. Review of the anaesthetic literature produced conflicting reports concerning the responses to neuromuscular blocking drugs in these patients. We describe a case in which vecuronium 0.11 mg kg(-1) produced prolonged neuromuscular blockade lasting 115 min in a patient with the condition. Conduction velocity in the facial nerve is usually less affected than the ulnar or peroneal nerve in CMT patients. This nerve may be more useful in monitoring neuromuscular blockade, both in titrating the dose of neuromuscular blocking drug and ensuring adequate reversal at the end of a procedure. Recent advances in molecular biology have enabled identification of the underlying genetic abnormalities and pathophysiology of CMT. These advances are reviewed and implications of CMT for the anaesthetist discussed. PMID- 11732533 TI - Severe unilateral bronchospasm mimicking inadvertent endobronchial intubation: a complication of the use of a topical lidocaine Laryngojet injector. AB - A healthy young woman is described in whom the left chest was unable to be inflated after intubation. The differential diagnosis and management are discussed. Severe unilateral bronchospasm was probably caused by topical lidocaine injected at the vocal cords and, inadvertently, into the left main bronchus with a Laryngojet device. PMID- 11732534 TI - Anaesthesia for Kartagener's syndrome. AB - Kartagener's syndrome is a hereditary syndrome involving a combination of dextrocardia (situs inversus), bronchiectasis and sinusitis, transmitted as an autosomal recessive trait. We describe a patient who had three anaesthetics over a period of a few months. Discussion relates to anaesthetic considerations in the syndrome and to recent findings relating to the molecular mechanisms of left right development. PMID- 11732535 TI - Acute right-to-left inter-atrial shunt; an important cause of profound hypoxia. AB - Three patients presented to our intensive care unit over a 3-yr period with profound hypoxia resulting from acute right-to-left inter-atrial shunt (RLIAS). Patient 1 was a 67-yr-old male with an atrial septal defect who became hypoxic and developed the rare sign of platypnoea following elective repair of an abdominal aortic aneurysm (breathlessness made worse when upright and relieved by lying flat). Patient 2 was a 38-yr-old female who developed platypnoea and hypoxia secondary to a patent foramen ovale (PFO) and pericardial effusion. Patient 3 was a 46-yr-old male with a PFO who developed hypoxia without platypnoea because of multiple pulmonary emboli following right hemicolectomy. These case reports illustrate the need to consider RLIAS as a cause of hypoxia of sudden onset. Early use of bubble contrast echocardiography is indicated. PMID- 11732536 TI - Air embolism during anaesthesia for shoulder arthroscopy. AB - We report a case of venous air embolism during an elective shoulder arthroscopy in which air was used as a joint distending agent. Venous air embolism was diagnosed by the sudden decrease in the end-tidal carbon dioxide concentration. The patient suffered no serious complications of venous air embolism and made a full recovery. We present this case to make surgeons and anaesthetists aware of the possibility of gas/air embolism during elective arthroscopy, when gas/air is used to distend the joint. This case also illustrates that the end-tidal carbon dioxide monitor, which is part of the standard anaesthetic monitoring system, is very sensitive in detecting venous air embolism. PMID- 11732537 TI - The inventions of John Blease. AB - Though he had no formal training in engineering, John Blease of Merseyside invented numerous devices that greatly benefited the practice of anaesthesia. Starting with the turning of component parts for simple anaesthetic machines in the 1930s, he was introduced to clinical anaesthesia and became skilled in the art of dental anaesthesia. In the early 1940s he developed the all-purpose Alfo Blease anaesthetic machine. In 1945 he designed an intermittent positive pressure ventilator, which was used successfully around Liverpool. After World War II he improved this into the Blease 'Pulmoflator', which was the first British positive pressure ventilator in commercial production. From then until the early 1960s he patented many other inventions, duly utilized in the manufacture of anaesthetic equipment, in which industry the Blease name survives in the company he founded. PMID- 11732538 TI - Serum S100 protein as a marker of cerebral damage during cardiac surgery. PMID- 11732539 TI - HIV exposure risk and management. PMID- 11732540 TI - Oral transmucosal fentanyl citrate as a premedicant. PMID- 11732541 TI - Effects of succinylcholine and rocuronium on the laryngeal neuromuscular junction. PMID- 11732542 TI - Fentanyl and midazolam anaesthesia for coronary bypass surgery. PMID- 11732543 TI - The management of major incidents. PMID- 11732544 TI - The management of major incidents. PMID- 11732545 TI - Assessing the reviewers of animal research. PMID- 11732546 TI - Assessing the reviewers of animal research. PMID- 11732547 TI - WHO ranking of health systems. PMID- 11732548 TI - Counting all species. PMID- 11732549 TI - But maybe counting is the easiest part. PMID- 11732550 TI - Research on resistance to cancer drug Gleevec. PMID- 11732551 TI - Propofol use in head-injury patients. PMID- 11732552 TI - Uses of error: and then there were three. PMID- 11732553 TI - Studies on the biosynthesis of the hydroxylysine-liked disaccharide unit of basement membranes and collagens. I. Kidney glucosyltransferase. PMID- 11732554 TI - Medicare program; changes to the hospital inpatient prospective payment systems and rates and costs of graduate medical education: fiscal year 2002 rates; provisions of the Balanced Budget Refinement Act of 1999; and provisions of the Medicare, Medicaid, and SCHIP Benefits Improvement and Protection Act of 2000. Final rules. AB - We are revising the Medicare hospital inpatient prospective payment systems for operating and capital costs to: implement applicable statutory requirements, including a number of provisions of the Medicare, Medicaid, and SCHIP [State Children's Health Insurance Program] Benefits Improvement and Protection Act of 2000 (Public Law 106-554); and implement changes arising from our continuing experience with these systems. In addition, in the Addendum to this final rule, we describe changes to the amounts and factors used to determine the rates for Medicare hospital inpatient services for operating costs and capital-related costs. These changes apply to discharges occurring on or after October 1, 2001. We also set forth the rate-of-increase limits as well as policy changes for hospitals and hospital units excluded from the prospective payment systems. We are making changes to the policies governing payments to hospitals for the direct costs of graduate medical education and critical access hospitals. Lastly, we are responding to public comments received on the following two related interim final rules that we published in the Federal Register and finalizing those interim rules: An August 1, 2000 interim final rule with comment period (65 FR 47026, HCFA-1131-IFC) that implemented, or conformed the regulations to, certain statutory provisions relating to Medicare payments to hospitals for inpatient services that were contained in the Medicare, Medicaid, and SCHIP Balanced Budget Refinement Act of 1999 (Public Law 106-113), and that were effective during FY 2000. These provisions related to reclassification of hospitals from urban to rural status, reclassification of certain hospitals for purposes of payment during fiscal year 2000, critical access hospitals, payments to hospitals excluded from the prospective payment system, and payments for indirect and direct graduate medical education costs. A June 13, 2001 interim final rule with comment period (66 FR 32172, HCFA-1178-IFC) that implemented, or conformed the regulations to, certain statutory provisions relating to Medicare payments to hospitals for inpatient services that were contained in Public Law 106- 554, and that were effective prior to passage of Public Law 106-554 on December 21, 2000; on April 1, 2001; or on July 1, 2001. Many of the provisions of Public Law 106 554 modified changes to the Social Security Act made by Public Law 106-113 or the Balanced Budget Act of 1997 (Public Law 105-33), or both. PMID- 11732555 TI - Child-resistant packaging for certain over-the-counter drug products. Final rule. AB - Pursuant to its 3-0 vote to do so, the Consumer Product Safety Commission (CPSC or Commission) is issuing a rule to require child-resistant (CR) packaging on drugs (OTC switched drugs) approved by the Food and Drug Administration (FDA) for over-the-counter (OTC) sale that contain active ingredients previously available only in prescription drugs. Current Commission regulations require CR packaging for most oral drug products containing prescription-only active ingredients.However, prior to issuance of this rule there was no general requirement to maintain CR packaging of such drug products in forms subsequently approved by the FDA for OTC sale. The Commission is also revoking the current prohibition on granting a petition for an exemption from a CR packaging requirement prior to FDA approval of the drug product in question. The Commission takes these actions under authority of the Poison Prevention Packaging Act of 1970, as amended. PMID- 11732556 TI - TRICARE; Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); eligibility and payment procedures for CHAMPUS beneficiaries age 65 and over. Interim final rule. AB - This interim final rule implements Section 712 of the Floyd D.Spence National Defense Authorization Act for Fiscal Year 2001. Section 712 extends TRICARE eligibility to persons age 65 and over who would otherwise have lost their TRICARE eligibility due to attainment of entitlement to hospital insurance benefits under Part A of Medicare. In order for these individuals to retain their TRICARE eligibility, they must be enrolled in the supplementary medical insurance program under Part B of Medicare. In general, in the case of medical or dental care provided to these individuals for which payment may be made under both Medicare and TRICARE, Medicare is the primary payer and TRICARE will normally pay the actual out-of-pocket costs incurred by the person. This rule prescribes TRICARE payment procedures and makes revisions to TRICARE rules to accommodate Medicare-eligible CHAMPUS beneficiaries. The Department is publishing this rule as an interim final rule in order to meet the statutorily required effective date. Public comments, however, are invited and will be considered when the rule is published as a final rule. PMID- 11732557 TI - Revisions to the requirements applicable to blood, blood components, and source plasma. Final rule. AB - The Food and Drug Administration (FDA) is amending the biologics regulations by removing, revising, or updating specific regulations applicable to blood, blood components, and Source Plasma to be more consistent with current practices in the blood industry and to remove unnecessary or outdated requirements. FDA is issuing this final rule as part of the agency's "Blood Initiative" in which FDA is reviewing and revising, when appropriate, its regulations, policies, guidance, and procedures related to blood, blood components, and Source Plasma. PMID- 11732558 TI - Statistical methods in epidemiology. VII. An overview of the chi2 test for 2 x 2 contingency table analysis. AB - PURPOSE: The odds ratio is an appropriate method of analysis for data in 2 x 2 contingency tables. However, other methods of analysis exist. One such method is based on the chi2 test of goodness-of-fit. Key players in the development of statistical theory include Pearson, Fisher and Yates. METHOD: Data are presented in the form of 2 x 2 contingency tables and a method of analysis based on the chi2 test is introduced. There are many variations of the basic test statistic, one of which is the chi2 test with Yates' continuity correction. The usefulness (or not) of Yates' continuity correction is discussed. Problems of interpretation when the method is applied to k x m tables are highlighted. RESULTS: Some properties of the chi2 the test are illustrated by taking examples from the author's teaching experiences. CONCLUSION: Journal editors should be encouraged to give both observed and expected cell frequencies so that better information comes out of the chi2 test statistic. PMID- 11732559 TI - Hemiplegic shoulder pain: defining the problem and its management. AB - PURPOSE: Hemiplegic shoulder pain can affect up to 70% of stroke patients and can have an adverse impact on rehabilitation outcomes. This article aims to review the literature on the suggested causes of hemiplegic shoulder pain and the therapeutic techniques that can be used to prevent or treat it. On the basis of this review, the components of an optimal management programme for hemiplegic shoulder pain are explored. METHOD: English language articles in the CINAHL and MEDLINE databases between 1990 and 2000 were reviewed. These were supplemented by citation tracking and manual searches. RESULTS: A management programme for hemiplegic shoulder pain could comprise the following components: provision of an external support for the affected upper limb when the patient is seated, careful positioning in bed, daily static positional stretches, motor retraining and strapping of the scapula to maintain postural tone and symmetry. CONCLUSIONS: Research is required to evaluate the effectiveness of the components of the proposed management programme for the prevention and treatment of hemiplegic shoulder pain and to determine in what combination they achieve the best outcomes. PMID- 11732560 TI - Living with arthritis--what is important? AB - PURPOSE: Demonstrating the effectiveness of health care interventions requires valid measurement of the impact of those interventions. However, outlining precisely what constitutes a 'good outcome' in the field of rehabilitation is no easy task and tends to rely on models proposed by 'experts' rather than people with the disabling conditions. This paper describes a study exploring outcomes that those people with a disabling condition (arthritis) consider important. METHOD: A qualitative study, interviewing 10 women with rheumatoid arthritis was carried out. The narratives were explored for categories and themes that encapsulated the perspective of the participants. RESULTS: A range of categories was identified and collated into five themes (personal/intrinsic factors, external/extrinsic factors, future issues, perceptions of normality and taking charge). CONCLUSIONS: The research supports in part, but also challenges more commonly used models of understanding the important consequences of disease and disability. The findings of the study may assist health professionals to reflect on current practice and reconsider processes used, and outcomes aimed for, in light of what patients/clients consider important. PMID- 11732561 TI - Predicting long-term care institution utilization among post-rehabilitation stroke patients in Taiwan: a medical centre-based study. AB - PURPOSE: Early identification of predictive factors relevant to the utilization of long-term care institution for stroke patients is important and thus investigated in this study on stroke patients receiving rehabilitation therapy. METHODS: This prospective follow-up investigation carried out during patients' clinical visits, at homes or long-term care institutions, was conducted at least 6 months after stroke on 151 stroke survivors. Functional ability was evaluated with the functional independence measure (FIM) instrument at discharge of the inpatient rehabilitation programme. Balance status was measured using the seven item balance scale of the Fugl-Meyer sensorimotor assessment (FMSA). Major medical, rehabilitative and sociodemographic factors were also examined during hospitalization period as independent variables. RESULTS: Of all the patients surveyed, 23 (15.2%) had been living in long-term care institutions. Univariate statistical analysis indicated that the significant factors related to long-term care institution utilization included recurrence of attack, bilateral involvement, impaired orientation, and functional and balance status at discharge. CONCLUSIONS: Basing on the significant predictors identified, analysis using the logistic regression model correctly classified three quarters of the subjects as long-term care institution residents. The strongest predictors of long-term care institution utilization for stroke patients following rehabilitation therapy were: bilaterally affected, impaired orientation and poor standing ability at discharge. PMID- 11732562 TI - Impairments, disabilities and handicap pattern in konzo--a non-progressive spastic para/tetraparesis of acute onset. AB - PURPOSE: To assess impairments, disabilities and handicap pattern in konzo. METHOD: The study included 17 konzo subjects, of which three were males and 14 females (mean age 21, median 18 years). A detailed neurological examination was performed on all subjects. Subsequently, an assessment of impairments, disabilities and handicap was done with a constructed rating scale partially based on the ICIDH-2 framework. RESULTS: The overall disablement picture in all subjects consisted of motor dysfunction in lower limbs leading to limitations in walking and movement activities, and restrictions in mobility. Hip mobility was severely impaired in most cases (15/17). Although konzo subjects showed normal muscle power in upper limbs (13/17), they had impaired fine motor function (10/17). CONCLUSION: Further studies are needed to assess the effectiveness of the WHO criteria for konzo in defining its forms. The applicability of the ICIDH 2 framework in this study demonstrates the possibility of its use as a common language among researchers in the field of motor disorders. However, a revision is suggested of its taxonomy, and a definition of operational criteria to clarify the content of different qualifiers provided to assess the level of functioning or disability. PMID- 11732563 TI - Screening for anxiety, depressive and somatoform disorders in rehabilitation- validity of HADS and GHQ-12 in patients with musculoskeletal disease. AB - PURPOSE: The detection of patients with comorbid mental illness is of high clinical importance in orthopaedic rehabilitation. To simplify detection of cases, screening instruments are recommended. The study investigated the discriminant validity of the Hospital Anxiety and Depression Scale (HADS) and the General Health Questionnaire (GHQ-12) to identify patients with comorbid mental disorders, specifically anxiety, depressive and somatoform disorders. METHOD: Two hundred and six patients with musculoskeletal disease from four orthopaedic rehabilitation clinics participated in a two-stage survey: (1) patients were assessed with the GHQ-12 and HADS; and (2) they were examined for DSM-IV mental disorders by clinical standardized interview (CIDI). Validity of the two instruments regarding the detection of mental disorders was compared using ROC analysis and CIDI-diagnoses as criteria. RESULTS: The HADS sumscale performed better in all analyses compared to the GHQ-12, specifically in detecting depressive and anxiety disorders. Best results are achieved for depressive disorders with an area under the curve (AUC) of 0.79, a sensitivity of 78% and a specificity of 71% (cut off point= 16). The positive predictive value (PPV) is best for the detection of any mental disorder with a cut-off point of 16 (46%). CONCLUSION: The HADS can be used as a screening instrument for the detection of comorbid depressive and anxiety disorders in patients with musculoskeletal disorders. Limitations in performance of screening instruments are due to: (1) different methodological approaches of tests (dimensional approach) and criterion (categorical approach); and (2) difficulties in diagnosing mental disorders in patients with prominent physical illness. PMID- 11732564 TI - Using a microswitch for vocalization responses with persons with multiple disabilities. AB - PURPOSE: This study was aimed at extending the evaluation of a special microswitch for vocalization responses with two persons with multiple disabilities. METHOD: The study was carried out according to an ABAB sequence (with A representing baseline and B treatment) with one person, and an AB sequence with the other person. Activation of the microswitch during treatment produced favourite environmental stimulation. RESULTS AND CONCLUSION: The results showed that both persons had an increase in the frequencies of vocalization responses during treatment. The importance of using these (fairly simple) responses within intervention programmes and some methodological issues were discussed. PMID- 11732565 TI - Apoptosis in preimplantation mammalian embryo and genetics. AB - The presence of dead cells in the preimplantation mammalian embryo has been well described. Since Kerr et al. (1972), it has become apparent that these cells die by apoptosis, a form of programmed cell death. This review analyses the recent morphological and biochemical evidence that apoptosis play a role in early mammalian embryo development. Normal and apoptotic (i.e. fragmented) embryos express several apoptosis-related genes during mammalian preimplantation embryo development, with severe changes when apoptosis is activated; these findings support a model in which mammalian preimplantation embryo development is regulated by the ratio of pro- and -anti- apoptotic genes. Apoptosis may be a normal feature in human preimplantation development, even in vivo, and may play an active role in the developing embryo through the removal of genetically abnormal cells. Contrary to these beneficial effects, apoptosis may have detrimental effects if either the number of apoptotic cells or ratio of these cells to the normal cells is elevated. According to this value, embryos could either continue to develop or arrest. PMID- 11732566 TI - Nucleolus formation in pre-implantation cattle and swine embryos. AB - Transcription of ribosomal RNA (rRNA) genes occurs in the nucleolus resulting in ribosome synthesis. In bovine and porcine embryos functional ribosome synthesizing nucleoli become structurally recognizable towards the end of the 4th and 3rd post-fertilization cell cycle, respectively. In cattle, a range of important nucleolar proteins become localized in the nucleolar anlage over several cell cycles and this localization is apparently completed towards the end of the 4th cell cycle. In swine, the localization of these proteins in the anlage is more synchronous and occurs towards the end of the 3rd cell cycle and is apparently completed at the onset of the 4th. The rRNA gene activation and the associated nucleolus formation may be used as a marker for the activation of the embryonic genome in mammalian embryos and, thus, serve to evaluate the developmental potential of embryos originating from different embryo technological procedures. PMID- 11732567 TI - Ultrastructural dynamic features of in vitro fertilization in humans. AB - The present study on in vitro fertilization in humans demonstrates that three dimensional (3-D) fine morphology by high resolution scanning electron microscopy (SEM) combined with parallel light and transmission electron microscopy (TEM) can reveal a number of new cellular detailed findings which cannot be detected with other methods. In this study the following aspects have been investigated in early human embryos. 1. Micro-topographical features of the zona pellucida (ZP), surface blastomeres and polar body. 2. Intracytoplasmic features of mature and healthy oocyte, in vitro fertilized (IVF) oocyte and early embryo development. 3. Comparison of general views of intracytoplasmic sperm injection (ICSI) and conventional IVF (C-IVF) of early embryos. 4. Presence of unusual large tubular smooth endoplasmic reticulum (SER) aggregates in 4, 5, and 6-cell embryos after ICSI. 5. Inside views of 3-D blastocysts such as inner cell mass and trophoblast. To our knowledge, this is the first time that such images are reported by using these techniques. PMID- 11732568 TI - Gene expression studies on human primordial germ cells and preimplantation embryos. AB - Research on the regulation of gene expression in human germ cells and preimplantation embryos is restricted due to the scarcity of samples and the requirement for highly sensitive molecular techniques to investigate the few cells available. To overcome these difficulties, we have developed a reliable procedure capable of generating amplified cDNA preparations from single cells. Using this procedure, we prepared cDNA from primordial germ cells (PGCs) isolated from the gonads of fetuses at 10 weeks gestation and from preimplantation embryos at the 1-cell, 4-cell, 8-cell and blastocyst stages. Our cDNA preparations allow us to investigate the expression profile of an almost unlimited number of different genes in the same sample preparation. This is of great advantage for studies of a panel of genes in a particular family or functional group, or with related mechanisms of regulation, e.g., developmental genes, oncogenes, cell cycle-control genes and imprinted genes. We have used these cDNA preparations in conjunction with differential display to identify genes specifically expressed in PGCs and preimplantation embryos in a sex- and developmental stage-specific manner. Genes specifically expressed in PGCs, oocytes and embryos were further analysed for their expression in embryonal carcinoma cells and in their differentiated derivatives following treatment by retinoic acid. Our strategy will disclose genes essential for gametogenesis and embryonic development which may only be expressed at certain stages of their development. The germ cell- and embryo-specific cDNA molecules, cDNA libraries and microarrays are a valuable resource for other researchers in this field. PMID- 11732569 TI - Experimental antigen modulation of blastocysts for mimicing dormant and invasive micrometastases. AB - The mouse preimplantation period can be prolonged experimentally by a delayed implantation, and this will keep the blastocyst in an inactive state. The dormancy can be interrupted by an oestrogen injection, which will make the blastocyst invasive. Thus the blastocyst mimics both dormant and invasive micrometastasis. Considering the similarities between blastocysts and micrometastases in cellular activation, we have developed methods for examining antigen modulations using the blastocyst as a model. Evaluating the total amount of antibody in the blastocyst and also the trophoblast antigen synthesis, shedding, and endocytosis, we found that dormant cells exposed to a surface specific antibody kept the immune complexes on the cell surface for a longer time than did the invasive cells. The rate of internalization of immune complexes was low in both dormant and invasive cells, but since the shedding activity was less active in the dormant cells they contained more antibodies totally than did the invasive ones under the same conditions. When exposing the cells to an anti cytoplamic antibody or to non-specific antibodies, the amount of antibodies in the cytoplasm of invasive cells was higher than in dormant cells. The methods used for examining the antigen modulation by the blastocyst should be useful also for studying the handling of antibodies by micrometastases. PMID- 11732570 TI - Ontogenesis of anti-oxidative enzymes in mouse embryos and fetuses: an immunohistochemical study. AB - The ontogenesis of two anti-oxidative enzymes, thioredoxin (TRX) and glutaredoxin (GRX), was examined immunohistochemically in mouse embryos and fetuses at various developmental stages. They were found to be localized in various tissues, with some tissue specificity and temporal sequence. Both TRX and GRX began to be expressed in many tissues at embryonic (E) day E10 or E11 and tended to increase as the developmental stage advanced. In the heart and neuroepithelium, however, their immunoreactivity was already positive at E8.5. In some fetal organs like the liver, pancreas and kidney, TRX and GRX showed heterogeneous localization, suggesting that their expression may reflect the variable functional states of the cell. These results suggest that TRX and GRX may be associated with tissue differentiation in embryos and fetuses are involved in the acquisition of the capacity of resistance against oxygen radicals. PMID- 11732571 TI - Is there a mesonephric cell contribution to the gonadal primordium before sexual differentiation in humans?: An ultrastructural study. AB - Differentiation of the intermediate mesoderm involves a complex series of events that result in the formation of the rudiment of the entire adult renal system, gonads and gonoducts. This work, using light, transmission, and scanning electron microscopy describes in human embryos of different ages, the development and spatiotemporal organization of the mesonephric nephron, and the development of the gonadal primordium, with the purpose of knowing if and how these two blastemas contribute to the origin of the non germinal cell content of the gonad primordium. Our results show that between Carnegie stage 13 and 20, the mesonephric nephrons facing the gonadal area, are separated by a band of mesenchymal tissue from the gonad primordium and they retain their structural integrities at the level of epithelial wall and their basement membrane. The morphological stability of basement membranes of different nephric structures, as well as the mesonephric duct during the period studied, confirm the previous opinion that the mesonephros is functional. During the same period of time, the structural events underlying gonad development show that primordial germ cells (PGCs) first invade the gonadal area, and thereafter interact both with epithelial coelomic cells and mesenchymal cells. Both types of cells surround PGCs, initiating the growth and differentiation of the gonadal primordium. Therefore, a mesonephric cell contribution to the genesis of the somatic cell components of the gonadal primordium should be discarded in humans. The present work emphasizes the need for more research in this field. PMID- 11732572 TI - The human embryo development through MMR. AB - This study evaluates the use of the Microscopic Magnetic Resonance (MMR) in the human prenatal development. Human embryos (8mm, 15mm, 18mm and 22mm in length) fixed in 4% paraformaldehyde were used. Results were compared with light microscopy (LM) images. The internal configuration of the embryos can be clearly observed as well as many organs such as liver, lungs, heart, including their spatial relationships. In general MMR sections are less clear and show minor details than those by LM. Neverthless, many advantages are provided by using this technique. For example it is possible: a) to make three-dimensional (3-D) surface and internal full or partial reconstructions; b) to evaluate the presence of developmental anomalies; c) to evaluate the tissular preservation degree of the specimens; and d) to apply morphometric techniques to unfixed specimens. In our opinion the advantages derived by using MMR are many and overcome the disadvantages. This study demonstrates that MMR can be incorporated into ordinary laboratory techniques in human development studies, being also an initial election technique opposite to others more aggressive. PMID- 11732573 TI - Ultrastructural morphodynamics of human Sertoli cells during testicular differentiation. AB - Our study reviews and ultrastructurally characterises human pre-Sertoli cells between the 6th and the 20th week of gestation by means of integrated light microscopy, transmission electron microscopy and high resolution scanning electron microscopy (standard or following ODO maceration). The morphofunctional differentiation of Sertoli cells defines testicular differentiation. These somatic cells are mostly of mesonephric origin and can be first morphologically recognised in 7 week-old embryos altogether with the formation of testicular cords. The latter organise as primordial germ cells surrounded by pre-Sertoli cells. Due to the great synthetic activity of pre-Sertoli cells the rough endoplasmic reticulum develops. The basal lamina of the cords becomes distinguishable at 7 to 8 weeks of gestation. Both prespermatogonia and pre Sertoli cells actively proliferate but the latter greatly outnumber prespermatogonia. Many interdigitations and cytoplasmic processes are observed between neighbouring pre-Sertoli cells. Due to cell proliferation a sort of compartmentalisation is established inside the cords in which pre-Sertoli cells tend to localise closer to the basal membrane embracing prespermatogonia with long and thin cytoplasmic processes. One of the main typical features of differentiating pre-Sertoli cells is the irregular nucleus and the prominent nucleolus. When the embryo is 14 to 20 weeks-old pre-Sertoli cells maintain their general morphology whereas the most significant change is the maximum development of Leydig cells. Testicular cords do not show any lumen at all so they cannot be termed "tubules". PMID- 11732575 TI - The sperm glycocalyx of Pezotettix giornai (Rossi) (Insecta: Orthoptera) after quick-freeze, deep-etching. AB - Mature spermatozoa of the catantopid orthopteran Pezotettix giornai are characterized by an elaborate external glycocalyx. This coating is removed during storage in spermatheca allowing preparation of sperm to their interaction with female gametes. We have studied this membrane coating in mature sperm by both conventional transmission electron microscopy and quick-freeze, deep-etching. The ultrastructural studies allowed visualization of three different domains in glycocalyx of testicular and deferent duct spermatozoa and only two in sperm cell isolated from seminal vesicles. These observations thus demonstrate that a remodeling of the male germ cell starts already during their storage in the seminal vesicle and is completed in the female genital tract. In this paper a three-dimensional (3D) model of sperm external coating is presented and discussed. PMID- 11732574 TI - Glial cell line-derived neurotrophic factor (GDNF) and its receptors GFRalpha-1 and GFRalpha-2 in the human testis. AB - Glial cell line-derived neurotrophic factor (GDNF) and its receptors GFRalpha-1 and GFRalpha-2 were found in the human testis during fetal development (15-34 weeks of gestation) and in adult men (51-86 years of age) by means of RT-PCR, immunohistochemistry and Western blot techniques. Gene expression of GDNF could be established in the human testis and immunoreactivity (IR) for GDNF was detectable in Leydig cells, Sertoli cells, some spermatocytes and round spermatids as well as in smooth muscle cells of the wall of arterioles and small arteries. In the adult human testis, Sertoli and Leydig cells showed GFRalpha-1 IR, whereas GFRalpha-2-IR was located exclusively in Leydig cells. Different to man, in the rat GDNF-IR in Sertoli cells was detectable only until postnatal day10, providing evidence for species related variability in the expression of GDNF. These findings suggest a critical role for GDNF during the differentiation of testicular structures and provide evidence for an additional important function in the adult human and rodent testis. PMID- 11732576 TI - Accumulation of transcripts in the mature human sperm nucleus: implication of the haploid genome in a functional role. AB - The existence of sperm-specific transcripts has been suggested by a number of studies performed both in man and rodents but their origin and role are not yet elucidated. For evaluating the functional significance of these mRNAs, transcripts coding for proteins expressed during spermiogenesis or potentially implicated in the early steps of zygote development, have been searched in human testis and sperm cells by RT-PCR. Furthermore their localization in spermatozoa has been checked by in situ hybridization. Our results confirm the presence of basic nucleoproteins (Transition proteins 1 and 2, Protamines 1 and 2) spermatozoal transcripts which probably represent remnants of previous transcription. They also reveal the existence of sperm specific mRNAs coding for the transcription factor Stat 4, the cyclin B1 and for the testicular isozyme of the angiotensin converting enzyme ACE. On the contrary, mRNAs coding for the heat shock protein Hsp 70 have been found in testis but not in spermatozoa. The possible roles of these transcripts either during the fertilization process or in zygote are discussed. PMID- 11732577 TI - Parathion induces mouse germ cells apoptosis. AB - Germ cell loss occurs in normal spermatogenesis at defined stages of the seminiferous epithelial cycle. The process has been known for over a century but only recently it was analyzed under the concept of apoptosis. This is a programmed cell death that occurs during development and also in the adult. It is believed to play a key role as quality control in sperm formation, avoiding the passage of genetic defects to future generations. Chemical toxicants may increase apoptosis, disturbing tissue homeostasis. The effect of the agropesticide parathion upon apoptosis in mouse seminiferous tubules was analyzed in young mice (onset of spermatogenesis) and in adult animals (full spermatogenesis). In both young and adult mice, the pesticide increases the rate of apoptosis, which takes place at stages where spermatogonial proliferation occurs, affects spermatocytes at the beginning of the meiotic process and spermatids at the elongating period. Basal apoptotic rates are greater in young mice. In adults, commercial parathion is more toxic than the pure organophosphoric compound. From these observations plus in vitro effects of parathion reported previously, it can be concluded that the pesticide affects DNA (and RNA and protein) synthesis. The effect is reversible with moderate doses of the chemical after acute intoxication. PMID- 11732578 TI - Microvasculature of the corpus epididymis of canis familiaris. A scanning electron microscopic study of microvascular corrosion casts. AB - The microvasculature of the corpus epididymis of adult domestic dogs (Canis familiaris) was studied using the technique of scanning electron microscopy of microvascular corrosion casts. The vascularization of the dog's corpus epididymis is accomplished by small arteries that course in the upper border of the organ. From these vessels arise first order arterioles (200-101 microm) which cross the surface of the epididymis and often show some coiling. The latter vessels supply second order arterioles (100-31 microm) which vascularize segments of the epididymal duct. The coiled and tight course of this duct is responsible for its peculiar microvascular arrangement. This is characterized by a 'bicycle-wheel' pattern and it is provided with precapillary or terminal arterioles (30-11 microm) nourishing a rich periductal network, mostly formed by anastomosing capillaries (10-3 microm). The periductal capillary network drains into postcapillary (11-30 microm), second order (31-100 microm) and first order (101 200 microm) venules, which empty into small calibre veins that course in the upper border of the epididymis. Anastomoses between vessels of the same type are often found. These are arteriovenous anastomoses mainly located at the periductal network. This study demonstrates that the epididymal microvasculature can be subdivided into a distribution microvascular network (DMN), formed by first and second order arterioles and venules, which presents a behaviour independent of the epididymal duct's sinuosities and a nutrition microvascular network (NMN), formed by precapillary arterioles, capillaries and post-capillary venules, characterised by the close relationship of the vessels with the loops of the epididymal duct. PMID- 11732579 TI - Multiple functions of mouse zona pellucida glycoprotein mZP3, the sperm receptor. AB - Zona pellucida glycoprotein mZP3 is a structural glycoprotein, a sperm receptor, and an acrosome reaction-inducer. The latter two functions are mediated, at least in part, by mZP3 oligosaccharides. mZP3 is unique to mammalian eggs, from mice to humans, although related glycoproteins are found in vitelline envelopes of a variety of eggs, from fish to birds. PMID- 11732581 TI - Estrogen as modulator of genes expression involved in control of reproduction. AB - The hypothalamic neurons, pituitary and their target cells, as genetically programmed to respond specifically to gonadal steroids (GS) and seasonal photoperiodic stimuli, play a crucial role in regulation of target genes transcription and regulatory ligands are conserved throughout evolution. Memory of these cells, acquired through the advanced evolution and programmed genes expression, may be modulated, transitory or permanently, by GS and estrogen (E) is a key hormone of these steroids. Pituitary chromophobes (Chr) and prolactin (PRL) cells reactions of birds and rats to GS and seasonal photoperiod were used as a parameter for elucidation of hypothalamic neurons and regulation of the animals reproduction. As a result of proliferative rate of chromophobes and hypertrophy of PRL cells, pituitary gonadotropic hormone (GTH) cells differentiation and activities are decreased, retarded or even inhibited and vice versa. The inverse relationship between PRL and GTH play a crucial role in regulation of reproduction. Light absorbed by rodopsin in the photoreceptor cells are transmitted via specific hypothalamic neurons to pars tuberalis cells, which are sensitive to photoperiodic stimuli and neurohormones as regulators of PRL cells activities. PMID- 11732580 TI - Expression of erythropoietin in human female reproductive organs. AB - Erythropoietin (Epo) is known to be a lineage specific cytokine which regulates the number of circulating erythrocytes. Most of it is produced in the kidney. Recently, Epo has been reported to be synthesized in the normal brain, placenta, and capillary endothelium. We also have found that uterine endometrium expresses Epo signals in an estrogen-dependent manner, and that Epo contributes to angiogenesis in the endometrium in mice. To clarify the functional activity of Epo in human reproductive organs, we examined Epo signaling in these organs by Southern analysis of RT-PCR products and studied the distribution of substances relevant to Epo signal transduction by immunohistochemistry and Western blotting. Epo mRNA is expressed in the normal human cervix, endometrium and ovary, but it is not always detected in the specimens. Immunohistochemical analysis revealed Epo-receptor (EpoR) protein in: a) the endothelium of vessels, in glandular and surface epithelial cells, in decidual cells of the endometrium, and b) in follicles at various stages including oocytes, granulosa, theca interna cells and lutein cells of the ovary. Moreover, co-expression of JAK2 and phosphotyrosine, which reflects tyrosine phosphorylation via JAK2, and co-expression of EpoR and STAT5, which is a transcriptional factor relevant to mitogenic activity, were seen at these Epo-responsive sites. Western blotting analysis of these organs confirmed the immunohistochemical results. These findings imply that female reproductive organs can produce Epo, and that signal transduction of Epo contributes to the cyclic changes in the female reproductive organs. PMID- 11732582 TI - Influence of estrogens and oxytocin on germ cells death in the neonatal mammalian ovary. AB - During mammalian oogenesis, some processes involve proliferation and others drastic reduction of germ cells. This study reports on the role played by two hormones, estradiol monobenzoate and oxytocin, in the control of the number of germ cells in the neonatal mouse ovary. Female neonatal mice were treated with doses ranging between 0.1 and 1 microg/mouse of estradiol monobenzoate or oxytocin and sacrificed at 5 days of postnatal age. The results showed that in the animals treated with estrogen, follicular development was more advanced than that of controls. Further the number of germ cells in apoptosis was drastically reduced. In the animals treated with oxytocin, the follicular development was arrested at the stage of primary follicles. In addition, the number of apoptotic germ cells increased if compared with that of the controls. PMID- 11732583 TI - Oocyte developmental competence after in vitro maturation depends on the persistence of cumulus-oocyte comunications which are linked to the intracellular concentration of cAMP. AB - Oocyte and cumulus cells are interconnected by an extensive network of gap junctions (GJ) formed by connexin 43. In the present study the functional and morphological status of GJ was investigated during in vitro maturation of bovine oocytes using media known to induce different rates of maturation and developmental competence as well as different levels of cumulus expansion and intracellular concentrations of cAMP. GJ functional condition was studied by microinjection of the fluorescent dye Lucyfer Yellow in cumulus-enclosed oocytes and the intracellular distribution of connexin 43 was examined by immunofluorescence immunocytochemistry. Both functional and morphological analysis of GJ between oocytes and cumulus cells indicated that high maturation and development rates are accompanied by the prolonged persistence of permeable communications, which, however, are independent from cumulus expansion. On the contrary, the premature interruption of such communications was linked to low maturation and development. When these results were correlated to the measurements of intracellular levels of cAMP it was observed that cumulus expansion, GJ permeability and good quality maturation require the highest levels of cAMP. Intermediate cAMP levels determine GJ permeability and good quality maturation in the absence of cumulus expansion while below a threshold level of cAMP, oocyte maturation is defective with a premature interruption of communications through GJ and lack of cumulus expansion which lead to poor development. PMID- 11732584 TI - Follicular development and ovulation in hypothyroid rdw rats. AB - In this paper, we summarized our recent studies on follicular development and ovulation in the presence or absence of thyroxine (T4) and equine chorionic gonadotrophin (eCG) treatment in infertile immature spontaneously hypothyroid rdw rats. T4 therapy once daily from postnatal day 21 to day 29 resulted in increases in ovarian weight on day 30 (P 0.01). Similar populations of both healthy and atretic antral follicles ranging in diameter from 101-400 microm were observed in control rdw and normal rats. T4 treatment markedly increased the numbers of healthy antral follicles of 101-400 microm or larger than 550 microm in diameter in the absence or presence of eCG, respectively, in rdw rats. On the other hand, T4 treatment did not affect the population of atretic antral follicles, but decreased the number of atretic large antral follicles (>400 microm) in the presence of eCG. Although a few (5+/-2) eggs were obtained from immature rdw rats treated with gonadotrophins alone, significantly more eggs (85+/-5) were ovulated from females treated with gonadotrophins and T4. As a control, normal littermates ovulated 45 eggs when treated with gonadotrophins only and 68 eggs when to them were also given T4. In conclusion, the results of the present study indicated that T4 treatment improved follicular development in the presence of eCG and significantly increased the number of ovulated eggs following hCG treatment in rdw rats. PMID- 11732585 TI - Histological characterization and in situ localization of apoptosis in the pig follicular atresia. AB - The aim of this study is the histological characterization of the atretic process of follicles in pig ovary. Two mechanisms have been found. In the atretic preantral follicles changes are produced in the oocyte, in the granulosa cells, an in the newly formed theca. Three histological phases can be distinguished during atresia in preantral follicles. The principal observed modification is the loss of the connections among granulosa cells that acquire a star-shaped aspect. Few apoptotic cells were observed. In the atretic antral follicles four histological phases were characterized. The presence of apoptotic cells increases progressively in the first three. A peculiar distribution of apoptotic cells in the granulosa as well as in the theca were observed in all phases. IN CONCLUSION: A) During atresia of antral follicles in porcine many apoptotic cells were observed in the granulosa and in the theca. B) Very few apoptotic cells were found during atresia in the preantral follicles. The possible existence of some other cell death mechanism different from apoptosis which may participate in the atresia of this type of follicles is proposed. PMID- 11732586 TI - Detailed morphology of human ovarian surface epithelium focusing on its metaplastic and neoplastic capability. AB - Ovarian epithelial tumors are thought to derive from the surface epithelium of the ovary, which differentiates after invagination of the coelomic mesothelium over the gonadal ridges during early embryonic development. On the putative mullerian potential of this epithelium, endometriosis, including ovarian endometriosis, can be explained by a metaplastic process from the peritoneum. In the present study, we assessed specific changes in the architecture and in the cytological characteristics of human ovarian epithelial cells on the surface and within the inclusion cyst. Grossly normal ovaries were retrieved from 215 patients undergoing incidental oophorectomy and 33 patients with contralateral ovarian tumor. In addition, 75 patients diagnosed with epithelial ovarian cancer and 26 ovarian endometriosis patients undergoing oophorectomy were selected. The incidence of cortical invagination, epithelial papillomatosis and pseudostratification was not different in normal ovaries between pre- and post menopausal women, whereas inclusion cysts were more frequently observed in post menopausal women (p<0.05). The occurrence rates of invagination and inclusion cysts were significantly higher in normal ovaries examined after incidental oophorectomy (p<0.05) and in those with contralateral ovarian tumor (p<0.05), respectively. Although serous metaplastic change with cilia was most common and it did not differ before and after menopause, the frequency of mucinous, endometrioid and transitional cell changes was low in the two groups of pre- and post-menopausal women. Mucinous metaplasia was more frequently observed in patients with contralateral ovarian tumor than those without ovarian tumor (12.1% vs. 1.9%; p<0.05). Four (5.3%) of 75 ovarian cancer patients revealed malignant transformed cells from a single layer of normal epithelium covering the ovarian surface or inclusion cyst. Ovarian or extraovarian endometriosis was identified in 16 (21.3%) of 75 epithelial ovarian cancer patients analyzed. An in situ carcinogenesis in the endometriotic cyst was seen in three epithelial ovarian cancers (4.0%). In seven (26.9%) of 26 ovarian endometriosis cases, epithelial cells on the ovarian surface or within the inclusion cyst were histologically changed to endometriotic gland cells. Careful and extensive observations of surgical specimens with ovarian carcinoma and ovarian endometriosis showed that they originated from the ovarian epithelium lining the surface and cortical small cysts. PMID- 11732587 TI - Histophysiology of the Fallopian tubes in relation to sperm binding, release, and completion of capacitation. AB - This paper is concerned with endosalpingeal specialisations that underlie stabilisation of unique secretions forming microenvironments within the Fallopian tube lumen, not least as they influence the final stages of sperm transport and maturation. In particular, a specific microenvironment at the ampullary-isthmic junction in the presence of the newly-shed oocyte(s), associated granulosa cells and their secretory products is thought to characterise the site of fertilisation. Attention is also drawn to the viscous glycoprotein in the caudal isthmus before ovulation: it isolates spermatozoa stored with suppressed motility from the metabolic stimulation caused by uterine or ampullary fluid. This viscous secretion strips male antigens from the sperm surface, facilitating their preovulatory binding reactions to organelles of the isthmus epithelium. Peri ovulatory activation of discrete numbers of viable spermatozoa is regulated by follicular progesterone secretion, but seemingly involves local mobilisation of Ca2+ ions into bound gametes of appropriate membranous maturity. Motility and progression to the site of fertilisation may be further heightened by an influence of catecholamines diffusing from the myosalpinx; receptors for such molecules are present on the sperm surface. There is tight control of sperm activation and release close to the time of ovulation, generating initial sperm:egg ratios at the ampullar-isthmic junction of close to unity. However, with establishment of the block to polyspermy in newly activated eggs and lapse of time after ovulation, control of sperm progression soon relaxes. Indeed, 2-4 cell embryos may contain large numbers of accessory spermatozoa on or in the zona pellucida. PMID- 11732588 TI - The regulatory function of the uterine epithelium for trophoblast attachment: experimental approaches. AB - Embryo implantation in the mammalian uterus is initiated by the formation of a direct cell-to-cell contact between the trophoblast of the blastocyst and the uterine epithelium. This process is far from trivial since apical plasma membranes of epithelial cells are normally non-adhesive. The uterine epithelium has the remarkable ability to enter, under steroid hormone control, a specific state ("receptivity") at which it can down-regulate this repellent property and can finally become apically adhesive for trophoblast (probably aided additionally by local paracrine signals). Experimental data from recent years are beginning to shed some light on the involved cell biological/molecular events. They will be discussed on the basis of concepts concerning the regulation of epithelial cell polarity and with side views on epithelial-mesenchymal transformation. Recently developed experimental in-vitro systems have allowed to detect a remarkable degree of selectivity in the interaction of trophoblast and uterine epithelium, in contrast to stroma invasion. A new approach enables us to determine actual adhesive forces between living trophoblast and uterine epithelial cells with a special modification of the atomic force microscope (force spectroscopy). The potential use of such an approach is discussed. PMID- 11732589 TI - Fertilization without spermatozoa. AB - Mammalian spermatozoa first acquire the ability to fertilize oocytes as they pass through the epididymis to mature. Due to recent advances in microinsemination techniques, not only mature spermatozoa, but also immature sperm cells at certain stages in the testis, have been used to construct diploid zygotes, some of which subsequently develop to normal offspring. Using round spermatids, the most youngest haploid male germ cells, normal births have been reported in the mouse, rabbit, and human. Furthermore, in the mouse, secondary and primary spermatocytes also support full term development after incorporation into immature or mature homologous oocytes. Spermatogenic cells of several species can be cryopreserved easily in simple cryoprotectant solutions. Thus, the microinsemination techniques using spermatogenic cells give us a way to treat infertility, and provide valuable information on gametogenesis, including spermatogenesis, meiosis, and genomic imprinting. PMID- 11732590 TI - The blood vasculature as the forming element of the uterus of the estrous donkey (Equus asinus). AB - Light, scanning electron microscopy of endometrial surface and vascular casts were used to study the vascular architecture of the donkey uterus during estrous. The arterial blood supply of the uterus comes from three arteries: the uterine branch of the ovarian artery, the uterine artery of the external iliac artery, and the uterine branch of the urogenital artery. All arteries enter the uterus at its mesometrial border and divide into smaller ones. Segmentally constricted arteries are seen to circumscribe large veins at the perimetrium which become highly convoluted in the intermuscular vascular layer of the myometrium. Small arteries and arterioles originate at the borderline between the myometrium and the endometrium and radiate to the surface of the endometrium to constitute a system of numerous ridges and grooves by a widely meshed plexus of subepithelial capillary network. The post-capillary venules of the endometrium arise from the subepithelial capillary plexus to form slightly larger veins than the concurrent arteries which join up to the large tortuous veins in the intermuscular vascular layer of the uterus. This arrangement of blood vessels in the donkey uterus and particularly in the endometrium provides the requirement for instant blood flow on the arterial side and for the slow flow rate on the venous side to ameliorate the process of substances exchange. PMID- 11732591 TI - Extracellular fibrillar matrix architecture of human placental villi at term. AB - The spatial organisation of the extracellular fibrillar matrix of normal human placental villi at term can be directly visualised by scanning electron microscopy after 2N-NaOH maceration technique. By these methods, the extracellular fibrillar matrix of placental villi appears as a continuous network of isolated collagen fibrils and/or small fibrillar bundles interwoven each other. This sort of "collagenous fibrillar skeleton" forms the axis of chorionic villi and connects them with the basal plates running through the whole villous system of the placenta. Significant variations in the spatial arrangement as well as in the quantity of the extracellular matrix is observed at different levels of the villous ramification. Within the stem villi, the fibrillar extracellular matrix are abundant and, whereas the fibrils near the villous surface run parallel to the longitudinal axis of the villous (outer fibrils), those located in the inner core of the villous are arranged circularly around the wall of the fetal vessels (inner fibrils). In mature intermediate and terminal villi, viceversa, the extracellular fibrillar matrix is significandy reduced and the fibrils are mainly organised in a thin circular layer around the capillaries and sinusoids. The present study demonstrated the existence of a diverse spatial architecture of the extracellular matrix that results to be peculiar to the various levels of the ramification of the villous tree. Therefore, these morphological data strongly suggest a "compartmentalisation" of the villous tree as suggested by previous immunohistochemical study. Such a highly organised "collagenous fibrillar skeleton" stresses the important mechanical role of the extracellular matrix in sustaining the chorionic fetal vessels and the trophoblastic layer. Furthermore, the fine reticular-meshed network observed within the terminal villi suggests that at this level an additional role ensuring a favourable milieu for active feto-maternal exchanges may exist. PMID- 11732592 TI - Syncytial units isolated from early human placentas. AB - This morphological study demonstrates that during the early phases of pregnancy, in the syncytium there are individual multinucleated units joined to one another by junctional complexes which probably are destined to flow together with the main syncytium by membrane breakdown. Normal human placentas from 6th to 12th week of pregnancy, derived from voluntary abortions, have been utilized. According to the method described by Tedde et al. (1991), clusters of villi have been isolated, washed in saline and subjected to a treatment with a 0.3% trypsin solution that caused the detachment of numerous cellular elements from the surface of the free villi. Such isolated elements have been subjected to centrifugation and separation in Percoll gradients. Then multinucleated syncytial units have been collected, attached to slides by cytocentrifugation, stained with toluidine blue, observed at the light microscope and utilized to count their nuclei: about 600 elements from different weeks of pregnancy have been considered and the results have been submitted to statistical analysis. The svncytial units are multinucleated and can often be different from one another, particularly in regard to the characteristics and the number of the nuclei. The evaluation of the number of nuclei contained in different syncytial elements shows that nuclei are less numerous in the early phases (6-8 weeks) and increase in number towards the end of the first trimester in a statistic significant way. In conclusion, the present study could confirm the existence of single syncytial units, joined to constitute the syncytial layer, probably destined to a complete fusion in the main syncytium and likely in functional relationship with the underlying Langhans cells. PMID- 11732593 TI - The human zona pellucida and scanning electron microscopy. Reality or artifacts? AB - The surface micro-morphology of the zona pellucida (ZP) was investigated in 158 inseminated but unfertilized mature human oocytes derived from assisted reproduction trials (ART) by means of traditional scanning electron microscopy (SEM) techniques (gold coating and conductive staining methods) and saponin ruthenium red-osmium tetroxide-thiocarbohydrazide method (Sap-RR-Os-TC). The main aspect of the ZP by traditional SEM (122 oocytes) consisted in a porous, net-like structure (97 oocytes), whereas a nearly smooth or compact structure of ZP was detected in 25 oocytes (79.5% vs 20.5%). Using Sap RR-Os-TC method on 36 oocytes, 31 oocytes showed ZP with alternating tight and large meshed networks, whereas 5 oocytes displayed only tight meshed network (86.1% vs 13.9%). Due to our well standardized procedures, to the stabilizing action of the conductive staining on the zona material and similar results obtained with the use of Sap RR-Os-TC method, we confidentially regard the ZP changes, occurring in oocytes of various groups, as genuine features, likely related to their actual maturation status, rather than as artifacts. In addition, we emphasize the concept that a modern view of the ZP surface implies the best evidence of crossing filaments' network. We think that the ZP "spongy" or "compact" appearance is only the result of microfilaments network collapse, not the true three-dimensional (3-D) representation of ZP structure. PMID- 11732594 TI - Regions responsible for organizing the microtubular cytoskeletons of extra embryonic membranes. AB - We have identified putative microtubule-organising centres in whole mounts and frozen sections of full term human placenta, amniochorion and first trimester chorionic villus tissue by indirect immunofluorescence confocal laser scanning microscopy. These sites may perform a crucial executive role in the morphogenesis of human extra-embryonic membranes, and also may establish polarity in the amniotic epithelium and mediate the branching pattern of growth essential to the production of the chorionic villus tree. PMID- 11732595 TI - Signal transduction pathways in activation of the mammalian egg. AB - The mature egg arrested at its second metaphase is a rather quiet cell, and possesses a pre-set developmental program. A sequence of biochemical events that leads to the rapid embryonic mitotic divisions initiates at fertilization when the spermatozoon overcomes this second cell cycle arrest. These events induce a transient rise in intracellular Ca2+ concentration ([Ca2+]i) that leads to the cortical granule exocytosis (CGE) and resumption of meiosis. Various treatments can induce parthenogenetic activation as manifested by the extrusion of the second polar body or CGE. Similar to somatic cells, recent studies in mammalian eggs suggest that signal transduction pathways mediate egg activation. The initial increase in ([Ca2+]i) appears to be critical for egg activation. Other messengers such as protein kinase C and protein tyrosine kinases, were suggested as possible inducers of some aspects of egg activation. PMID- 11732596 TI - Fertilization imaged in 2-, 3- and four dimensions: molecular insights for treating infertility. AB - Fertilization in humans follows a complex series of events including binding of the sperm to the oocyte plasma membrane, oocyte activation, the completion of meiotic maturation of the oocyte with the extrusion of the second polar body, the decondensation of the sperm nucleus and the maternal chromosomes into male and female pronuclei and the restoration of the sperm centrosome. This duplicates and separates, forming two mitotic spindle poles upon which the parental genomes can intermix to complete fertilization. The use of intracytoplasmic sperm injection (ICSI) has been highly effective as a treatment for severe male infertility and thousands of ICSI babies have been born world-wide. Working with rhesus monkey gametes, we have developed a preclinical animal model for understanding the cell biological basis of ICSI. Typically, ICSI results in abnormal nuclear remodeling during sperm decondensation due to the presence of the sperm acrosome and perinuclear structures normally removed at the oolemma during in vitro fertilization. These unusual modifications raise concerns that the ICSI procedure itself might lead to the observed increase in chromosome anomalies reported for PMID- 11732597 TI - Characterization of human gamete centrosomes for assisted reproduction. AB - Centrosomes of human gametes need to be characterised in more detail, since it was recently shown in 1991 that the human embryo inherits the dominant paternal centrosome at fertilization. Thus there has been a resurgence of centrosomal research in the last decade. The sperm centrosome, containing a single centriole, is preserved and dominant while the egg centrosome is reduced and inactivated during oogenesis, since there should be only one functional centrosome to ensure normal development. This presentation reviews the ultrastructure of gamete centrosomes and attempts to define their respective roles in assisted fertilization. Human testicular and ovarian tissues, sperm, eggs and zygotes were examined by routine TEM. Developing sperm cells and oogonia have two functional centrioles (diplosomes) in their centrosomes, showing the typical "9+0" organisation of microtubule triplets--common to somatic cells. The diplosomes are surrounded by pericentriolar material (PCM), which nucleate microtubules (MT) that organize the cytoskeleton and mitotic spindles. During spermiogenesis, when the spermatids transform into mature sperm, there is a partial reduction of the male centrosome, during which the proximal centriole (PC) is retained intact in the sperm neck, proximal to the nucleus, while the distal centriole (DC) which gave rise to the sperm flagellum, is partially reduced and merges distally with the sperm axoneme in the midpiece and tail. The proximal functional centriole is located in a "black box" in the neck, composed of the capitulum beneath the basal plate and flanked laterally by 9 segmented columns and shows the typical "9+0" organisation of triplets of MT. The PC contains dense material both within and outside the triplets, and the central doublet of axonemal MT terminates in a clump of dense material below its lower vault. The DC is disorganised proximally showing loss of triplets and cannot function as a typical centriole, since the central doublets of axonemal MT traverse through it. Oogonia present a pair of well-defined centrioles, which are involved in cell division. These are lost during oogenesis and the mature oocyte is devoid of centrioles, as in most mammals. Neither does the human oocyte have granular centrosomal material at meiotic spindle poles, in contrast to mouse oocytes which have a dominant maternal centrosome. Thus the oocyte centrosome is greatly reduced and inactivated. Functional centrosomal structure is, however, restored after fertilization in the zygote with some maternal input around the sperm centriole, which duplicates at the pronuclear stage, forms a sperm aster and proceeds to form the first mitotic spindle. This is the ancestor of centrosomes in embryonic, foetal and adult somatic cells. In 1991 (Sathananthan, 1991), we postulated that sperm centrosomal dysfunction could lead to aberrant embryonic development based on centriolar defects in sperm with impaired motility. This hypothesis has now gained acceptance and further evidence to support this theory of infertility is presented. PMID- 11732598 TI - In vitro fertilized human eggs. An electron microscopic study. AB - This paper describes by scanning and transmission electron microscopy the ultrastructure of the human fertilized egg and its vestments (cumulus oophorus and zona pellucida). Data are reported on the ultrastructure of a. conventional in vitro fertilized eggs (pronuclear eggs and cleaving eggs at two-to-four cell stage); b. eggs at the same developmental stage deriving fro intracytoplasmic sperm injection. The present results showed that: 1. The cumulus-enclosed fertilized egg is a highly dynamic structure in which egg vestments play a crucial role, positively affecting fertilization and healthy embryo development; 2. Intracytoplasmic sperm injection technique does not seem to significantly alter fertilized egg morphology. PMID- 11732599 TI - Regulatory systems in early mammalian development, with especial reference to polarity and totipotency. AB - Increasing evidence is now available to reveal some of the underlying regulatory systems regulating early mammalian embryogenesis. These include timing, integration, multipotency, polarities and other embryological systems. This paper deals mostly with polarities and axes, and their integration with other regulatory phenomena. Various morphogenetic aspects of ovulated oocytes and cleavage planes have long been suspected as displaying gradients, and events such as ooplasmic rotation at sperm entry, the imposition of nucleolar polarity and the highly regulated cleavage planes add to this evidence. Examples of proteins that form gradients according to the embryonic axes are increasing in number, and provide molecular evidence of their nature in establishing a persistent polarity throughout early cleavage stages. The same polarised systems are also involved in the blastocyst and implantation, and their characteristics influence implantation and the post-implantation growth of the embryo. More knowledge on each of these major regulatory systems is needed to clarify the exact nature of early mammalian differentiation and organogenesis. PMID- 11732600 TI - Robert G. Edwards and Ryuzo Yanagimachi and the development of modern embryology and human reproduction. PMID- 11732601 TI - Understanding evolution. PMID- 11732602 TI - The left-handed double helical nucleic acids. AB - The conversion of right-handed dsDNA and dsRNA to the left-handed Z-conformation involves a reorganization of the nucleotides relative to each other. This conversion can be facilitated by the tight binding of a Z-conformation-specific protein domain from the editing enzyme dsRNA adenosine deaminase. This may influence the modification of both pre-mRNAs as well as some replicating RNA viruses. PMID- 11732603 TI - Genomics and the evolution of aminoacyl-tRNA synthesis. AB - Translation is the process by which ribosomes direct protein synthesis using the genetic information contained in messenger RNA (mRNA). Transfer RNAs (tRNAs) are charged with an amino acid and brought to the ribosome, where they are paired with the corresponding trinucleotide codon in mRNA. The amino acid is attached to the nascent polypeptide and the ribosome moves on to the next codon. Thus, the sequential pairing of codons in mRNA with tRNA anticodons determines the order of amino acids in a protein. It is therefore imperative for accurate translation that tRNAs are only coupled to amino acids corresponding to the RNA anticodon. This is mostly, but not exclusively, achieved by the direct attachment of the appropriate amino acid to the 3'-end of the corresponding tRNA by the aminoacyl tRNA synthetases. To ensure the accurate translation of genetic information, the aminoacyl-tRNA synthetases must display an extremely high level of substrate specificity. Despite this highly conserved function, recent studies arising from the analysis of whole genomes have shown a significant degree of evolutionary diversity in aminoacyl-tRNA synthesis. For example, non-canonical routes have been identified for the synthesis of Asn-tRNA, Cys-tRNA, Gln-tRNA and Lys-tRNA. Characterization of non-canonical aminoacyl-tRNA synthesis has revealed an unexpected level of evolutionary divergence and has also provided new insights into the possible precursors of contemporary aminoacyl-tRNA synthetases. PMID- 11732604 TI - The fidelity of the translation of the genetic code. AB - Aminoacyl-tRNA synthetases play a central role in maintaining accuracy during the translation of the genetic code. To achieve this challenging task they have to discriminate against amino acids that are very closely related not only in structure but also in chemical nature. A 'double-sieve' editing model was proposed in the late seventies to explain how two closely related amino acids may be discriminated. However, a clear understanding of this mechanism required structural information on synthetases that are faced with such a problem of amino acid discrimination. The first structural basis for the editing model came recently from the crystal structure of isoleucyl-tRNA synthetase, a class I synthetase, which has to discriminate against valine. The structure showed the presence of two catalytic sites in the same enzyme, one for activation, a coarse sieve which binds both isoleucine and valine, and another for editing, a fine sieve which binds only valine and rejects isoleucine. Another structure of the enzyme in complex with tRNA showed that the tRNA is responsible for the translocation of the misactivated amino-acid substrate from the catalytic site to the editing site. These studies were mainly focused on class I synthetases and the situation was not clear about how class II enzymes discriminate against similar amino acids. The recent structural and enzymatic studies on threonyl-tRNA synthetase, a class II enzyme, reveal how this challenging task is achieved by using a unique zinc ion in the active site as well as by employing a separate domain for specific editing activity. These studies led us to propose a model which emphasizes the mirror symmetrical approach of the two classes of enzymes and highlights that tRNA is the key player in the evolution of these class of enzymes. PMID- 11732606 TI - MADS-box genes are involved in floral development and evolution. AB - MADS-box genes encode transcription factors in all eukaryotic organisms thus far studied. Plant MADS-box proteins contain a DNA-binding (M), an intervening (I), a Keratin-like (K) and a C-terminal C-domain, thus plant MADS-box proteins are of the MIKC type. In higher plants most of the well-characterized genes are involved in floral development. They control the transition from vegetative to generative growth and determine inflorescence meristem identity. They specify floral organ identity as outlined in the ABC model of floral development. Moreover, in Antirrhinum majus the MADS-box gene products DEF/GLO and PLE control cell proliferation in the developing flower bud. In this species the DEF/GLO and the SQUA proteins form a ternary complex which determines the overall "Bauplan" of the flower. Phylogenetic reconstructions of MADS-box sequences obtained from ferns, gymnosperms and higher eudicots reveal that, although ferns possess already MIKC type genes, these are not orthologous to the well characterized MADS box genes from gymnosperms or angiosperms. Putative orthologs of floral homeotic B- and C-function genes have been identified in different gymnosperms suggesting that these genes evolved some 300-400 million years ago. Both gymnosperms and angiosperms also contain a hitherto unknown sister clade of the B-genes, which we termed Bsister. A novel hypothesis will be described suggesting that B and Bsister might be involved in sex determination of male and female reproductive organs, respectively. PMID- 11732605 TI - Methionyl-tRNA synthetase. AB - Methionyl-tRNA synthetase (MetRS) belongs to the family of 20 enzymes essential for protein biosynthesis. It links covalently methionine with its cognate tRNA. Crystal structures solved for bacterial MetRSs have given a number of interesting insights into enzyme architecture and methionylation catalysis. A comparison of sequences of MetRSs belonging to all kingdoms of life, as well as numerous biochemical and genetic studies have revealed the presence of various additional domains appended to the catalytic core of synthetase. They are responsible for interactions with tRNA and proteins. Tertiary structure of C-terminal tRNA binding appendices can be deduced from those determined for their homologues: tRNA binding protein 111 and endothelial monocyte-activating polypeptide II. Contacts between MetRS and other proteins could be mediated not only by noncatalytic peptides but also by structural elements present in the catalytic core, e.g. Arg-Gly-Asp (RGD) motifs. Additional activities involve MetRS in the maintenance of translational fidelity and in coordination of ribosome biogenesis with protein synthesis. PMID- 11732607 TI - Nod genes and Nod signals and the evolution of the Rhizobium legume symbiosis. AB - The establishment of the nitrogen-fixing symbiosis between rhizobia and legumes requires an exchange of signals between the two partners. In response to flavonoids excreted by the host plant, rhizobia synthesize Nod factors (NFs) which elicit, at very low concentrations and in a specific manner, various symbiotic responses on the roots of the legume hosts. NFs from several rhizobial species have been characterized. They all are lipo-chitooligosaccharides, consisting of a backbone of generally four or five glucosamine residues N acylated at the non-reducing end, and carrying various O-substituents. The N-acyl chain and the other substituents are important determinants of the rhizobial host specificity. A number of nodulation genes which specify the synthesis of NFs have been identified. All rhizobia, in spite of their diversity, possess conserved nodABC genes responsible for the synthesis of the N-acylated oligosaccharide core of NFs, which suggests that these genes are of a monophyletic origin. Other genes, the host specific nod genes, specify the substitutions of NFs. The central role of NFs and nod genes in the Rhizobium-legume symbiosis suggests that these factors could be used as molecular markers to study the evolution of this symbiosis. We have studied a number of NFs which are N-acylated by alpha,beta unsaturated fatty acids. We found that the ability to synthesize such NFs does not correlate with taxonomic position of the rhizobia. However, all rhizobia that produce NFs such nodulate plants belonging to related tribes of legumes, the Trifolieae, Vicieae, and Galegeae, all of them being members of the so-called galegoid group. This suggests that the ability to recognize the NFs with alpha beta-unsaturated fatty acids is limited to this group of legumes, and thus might have appeared only once in the course of legume evolution, in the galegoid phylum. PMID- 11732608 TI - Reduction of bacterial genome size and expansion resulting from obligate intracellular lifestyle and adaptation to soil habitat. AB - Prokaryotic organisms are exposed in the course of evolution to various impacts, resulting often in drastic changes of their genome size. Depending on circumstances, the same lineage may diverge into species having substantially reduced genomes, or such whose genomes have undergone considerable enlargement. Genome reduction is a consequence of obligate intracellular lifestyle rendering numerous genes expendable. Another consequence of intracellular lifestyle is reduction of effective population size and limited possibility of gene acquirement via lateral transfer. This causes a state of relaxed selection resulting in accumulation of mildly deleterious mutations that can not be corrected by recombination with the wild type copy. Thus, gene loss is usually irreversible. Additionally, constant environment of the eukaryotic cell renders that some bacterial genes involved in DNA repair are expandable. The loss of these genes is a probable cause of mutational bias resulting in a high A+T content. While causes of genome reduction are rather indisputable, those resulting in genome expansion seem to be less obvious. Presumably, the genome enlargement is an indirect consequence of adaptation to changing environmental conditions and requires the acquisition and integration of numerous genes. It seems that the need for a great number of capabilities is common among soil bacteria irrespective of their phylogenetic relationship. However, this would not be possible if soil bacteria lacked indigenous abilities to exchange and accumulate genetic information. The latter are considerably facilitated when housekeeping genes are physically separated from adaptive loci which are useful only in certain circumstances. PMID- 11732609 TI - Editing of plant mitochondrial transfer RNAs. AB - Editing in plant mitochondria consists in C to U changes and mainly affects messenger RNAs, thus providing the correct genetic information for the biosynthesis of mitochondrial (mt) proteins. But editing can also affect some of the plant mt tRNAs encoded by the mt genome. In dicots, a C to U editing event corrects a C:A mismatch into a U:A base-pair in the acceptor stem of mt tRNAPhe (GAA). In larch mitochondria, three C to U editing events restore U:A base-pairs in the acceptor stem, D stem and anticodon stem, respectively, of mt tRNAHis (GUG). For both these mt tRNAs editing of the precursors is a prerequisite for their processing into mature tRNAs. In potato mt tRNACys (GCA), editing converts a C28:U42 mismatch in the anticodon stem into a U28:U42 non-canonical base-pair, and reverse transcriptase minisequencing has shown that the mature mt tRNACys is fully edited. In the bryophyte Marchantia polymorpha this U residue is encoded in the mt genome and evolutionary studies suggest that restoration of the U28 residue is necessary when it is not encoded in the gene. However, in vitro studies have shown that neither processing of the precursor nor aminoacylation of tRNACys requires C to U editing at this position. But sequencing of the purified mt tRNACys has shown that psi is present at position 28, indicating that C to U editing is a prerequisite for the subsequent isomerization of U into psi at position 28. PMID- 11732610 TI - How RNA viruses exchange their genetic material. AB - One of the most unusual features of RNA viruses is their enormous genetic variability. Among the different processes contributing to the continuous generation of new viral variants RNA recombination is of special importance. This process has been observed for human, animal, plant and bacterial viruses. The collected data reveal a great susceptibility of RNA viruses to recombination. They also indicate that genetic RNA recombination (especially the nonhomologous one) is a major factor responsible for the emergence of new viral strains or species. Although the formation and accumulation of viral recombinants was observed in numerous RNA viruses, the molecular basis of this phenomenon was studied in only a few viral species. Among them, brome mosaic virus (BMV), a model (+)RNA virus offers the best opportunities to investigate various aspects of genetic RNA recombination in vivo. Unlike any other, the BMV-based system enables homologous and nonhomologous recombination studies at both the protein and RNA levels. As a consequence, BMV is the virus for which the structural requirements for genetic RNA recombination have been most precisely established. Nevertheless, the previously proposed model of genetic recombination in BMV still had one weakness: it could not really explain the role of RNA structure in nonhomologous recombination. Recent discoveries concerning the latter problem give us a chance to fill this gap. That is why in this review we present and thoroughly discuss all results concerning nonhomologous recombination in BMV that have been obtained until now. PMID- 11732611 TI - Ribozymes of the hepatitis delta virus: recent findings on their structure, mechanism of catalysis and possible applications. AB - Although the delta ribozymes have been studied for more than ten years the most important information concerning their structure and mechanism of catalysis were only obtained very recently. The crystal structure of the genomic delta ribozyme turns out to be an excellent example of the extraordinary properties of RNA molecules to fold into uniquely compact structures. Details of the X-ray structure have greatly stimulated further studies on the folding of the ribozymes into functionally active molecules as well as on the mechanism of RNA catalysis. The ability of the delta ribozymes to carry out general acid-base catalysis by nucleotide side chains has been assumed in two proposed mechanisms of self cleavage. Recently, considerable progress has been also made in characterizing the catalytic properties of trans-acting ribozyme variants that are potentially attractive tools in the strategy of directed RNA degradation. PMID- 11732612 TI - Structure-function relationship of serine protease-protein inhibitor interaction. AB - We report our progress in understanding the structure-function relationship of the interaction between protein inhibitors and several serine proteases. Recently, we have determined high resolution solution structures of two inhibitors Apis mellifera chymotrypsin inhibitor-1 (AMCI-I) and Linum usitatissimum trypsin inhibitor (LUTI) in the free state and an ultra high resolution X-ray structure of BPTI. All three inhibitors, despite totally different scaffolds, contain a solvent exposed loop of similar conformation which is highly complementary to the enzyme active site. Isothermal calo- rimetry data show that the interaction between wild type BPTI and chymotrypsin is entropy driven and that the enthalpy component opposes complex formation. Our research is focused on extensive mutagenesis of the four positions from the protease binding loop of BPTI: P1, P1', P3, and P4. We mutated these residues to different amino acids and the variants were characterized by determination of the association constants, stability parameters and crystal structures of protease-inhibitor complexes. Accommodation of the P1 residue in the S1 pocket of four proteases: chymotrypsin, trypsin, neutrophil elastase and cathepsin G was probed with 18 P1 variants. High resolution X-ray structures of ten complexes between bovine trypsin and P1 variants of BPTI have been determined and compared with the cognate P1 Lys side chain. Mutations of the wild type Ala16 (P1') to larger side chains always caused a drop of the association constant. According to the crystal structure of the Leu16 BPTI-trypsin complex, introduction of the larger residue at the P1' position leads to steric conflicts in the vicinity of the mutation. Finally, mutations at the P4 site allowed an improvement of the association with several serine proteases involved in blood clotting. Conversely, introduction of Ser, Val, and Phe in place of Gly12 (P4) had invariably a destabilizing effect on the complex with these proteases. PMID- 11732613 TI - Nucleoside phosphate analogues of biological interest, and their synthesis via aryl nucleoside H-phosphonates as intermediates. AB - This review presents a brief account of the chemistry and mechanistic aspects of aryl H-phosphonates, and selected applications of this class of compounds as intermediates in the synthesis of a wide range of biologically important analogues of nucleoside phosphates, and oligonucleotides, in which the phosphate moieties are replaced by other structurally related groups. The aryl nucleoside H phosphonates, compounds of controlled reactivity, have proven to be more versatile and superior to various mixed anhydrides as synthetic intermediates, particularly for preparation of nucleotide analogues bearing P-N or P-S bonds in various configurational arrangements at the phosphate moiety. PMID- 11732614 TI - Organismal view of a plant and a plant cell. AB - Cell walls are at the basis of a structural, four-dimensional framework of plant form and growth time. Recent rapid progress of cell wall research has led to the situation where the old, long-lasting juxtaposition: "living" protoplast--"dead" cell wall, had to be dropped. Various attempts of re-interpretation cast, however, some doubts over the very nature of plant cell and the status of the walls within such a cell. Following a comparison of exocellular matrices of plants and animals, their position in relation to cells and organisms is analysed. A multitude of perspectives of the biological organisation of living beings is presented with particular attention paid to the cellular and organismal theories. Basic tenets and resulting corollaries of both theories are compared, and evolutionary and developmental implications are considered. Based on these data, "The Plant Body"--an organismal concept of plants and plant cells is described. PMID- 11732615 TI - The cell-free protein biosynthesis--applications and analysis of the system. AB - The in vitro protein biosynthesis has the potentials to become a powerful technology for biochemical research. Beside the determination of structure and function the in vitro evolution of proteins is also of great interest. The system described was used to produce bovine heart fatty acid binding protein (FABP) and bacterial chloramphenicol acetyltransferase (CAT) with and without fusion of the Strep-tag II affinity peptide. The proteins were purified after and during protein biosynthesis by using a StrepTactin Sepharose matrix. No significant influence of the Strep-tag and the conditions during the affinity chromatography on maturation or activity of the protein was observed. The in vitro evolution of proteins is feasible by means of ribosome display. The selection of a specific mRNA coding for a shortened FABP with a N-terminal His-tag via the accompanying protein property was shown. Goal of the selection was to bind the FABP via the His-tag on Ni(II)-IDA-agarose. After nine cycles of transcription, translation, affinity selection and RT-PCR the protein with the His-tag could be enriched 10(8)-fold. In order to correlate a possible relationship between changes in protein population and biological function studies were initiated in which 2 dimensional protein patterns of the total in vitro system were compared after 0 and 2 h reaction time. The very interesting findings are that a number of proteins disappear, while others are newly formed during protein synthesis. PMID- 11732616 TI - Genetic variability of potato spindle tuber viroid RNA replicon. AB - The genetic continuity of the potato spindle tuber viroid (PSTVd) genome was analysed after infection of tomato plants with cloned cDNAs of parental strains. During the six weeks of the experiment, several new sequence variants appeared. The sequence variants detected in the progeny population induced sequence specific disease symptoms. The PSTVd genome therefore follows the pattern expected for typical pseudo-strains propagating in plants as a population of similar sequences. Assessing further the replicon continuity, a PSTVd cDNA mutant with a deletion in the central conserved region was constructed and proven to be non-infectious. Surprisingly, in a sub-population of potato transformants expressing the same deleted PSTVd RNA an infectious viroid was detected. This suggests specific transcript conversion followed by recovery of the full-length pathogen genome. PMID- 11732617 TI - Sequence determination and analysis of S-adenosyl-L-homocysteine hydrolase from yellow lupine (Lupinus luteus). AB - The coding sequences of two S-adenosyl-L-homocysteine hydrolases (SAHases) were identified in yellow lupine by screenig of a cDNA library. One of them, corresponding to the complete protein, was sequenced and compared with 52 other SAHase sequences. Phylogenetic analysis of these proteins identified three groups of the enzymes. Group A comprises only bacterial sequences. Group B is subdivided into two subgroups, one of which (B1) is formed by animal sequences. Subgroup B2 consist of two distinct clusters, B2a and B2b. Cluster B2b comprises all known plant sequences, including the yellow lupine enzyme, which are distinguished by a 50-residue insert. Group C is heterogeneous and contains SAHases from Archaea as well as a new class of animal enzymes, distinctly different from those in group B1. PMID- 11732618 TI - Elements of thermodynamics in RNA evolution. AB - The paper presents some aspects correlating thermal stability of RNA folding and the occurrence of structural motifs in natural ribonucleic acids. Particularly, the thermodynamic stability of 2'-5' and 3'-5' linked RNA and the contribution of unpaired terminal nucleotides (dangling ends) in secondary (2D) and tertiary (3D) structures of RNA are discussed. Both examples suggest that during evolution nature selected sequences and structures of RNA which are the most thermally stable and efficient for their biological function. PMID- 11732619 TI - Mg2+ ions do not induce expansion of the melted DNA region in the open complex formed by Escherichia coli RNA polymerase at a cognate synthetic Pa promoter. A quantitative KMnO4 footprinting study. AB - Footprinting studies of prokaryotic open transcription complexes (RPO), based on oxidation of pyrimidine residues by KMnO4 and/or OsO4 at a single oxidant dose, have suggested that the extent of DNA melting in the transcription bubble region increases in the presence of Mg . In this work, quantitative KMnO4 footprinting in function of the oxidant dose of RPO, using Escherichia coli RNA polymerase (E(sigma)70) at a fully functional synthetic promoter Pa having -35 and -10 consensus hexamers, has been used to determine individual rate constants of oxidation of T residues in this region at 37degrees C in the absence of Mg2+ and in the presence of 10 mM MgCl2, and to evaluate therefrom the effect of Mg2+ on the extent of DNA melting. Population distributions of end-labeled DNA fragments corresponding to oxidized Ts were quantified and analyzed according to the single hit kinetic model. Pseudo-first order reactivity rate constants, ki, thus obtained demonstrated that Mg2+ ions bound to RPO merely enhanced the reactivity of all 11 oxidizable thymines between the +3 and -11 promoter sites by a position dependent factor: 3-4 for those located close to the transcription start point +1 in either DNA strand, and about 1.6 for those located more distantly therefrom. On the basis of these observations, we conclude that Mg2+ ions bound to RPO at Pa do not influence the length of the melted DNA region and propose that the higher reactivity of thymines results mainly from lower local repulsive electrostatic barriers to MnO4 diffusion around carboxylate binding sites in the catalytic center of RPO and promoter DNA phosphates. PMID- 11732620 TI - Effect of Mg2+ on kinetics of oxidation of pyrimidines in duplex DNA by potassium permanganate. AB - Potassium permanganate oxidation of pyrimidine bases is often used to probe single-stranded regions in functional DNA-protein complexes. However, so far reactivity of these bases in double-stranded DNA has not been studied quantitatively. We have investigated the kinetics of oxidation of pyrimidines in supercoiled pDS3 plasmid dsDNA by quantitative KMnO4 footprinting, in connection with parallel studies on the effect of Mg2+ on kinetics of oxidation of individual thymines in the single-stranded region of the open transcription complex of Escherichia coli RNA polymerase at a cognate Pa promoter contained in this plasmid. Rate constants of oxidation for pyrimidines, kj, in selected regions of pDS3 DNA, including Pa promoter, were determined under single-hit reaction conditions in the absence and presence of 10 mM MgCl2. Their values appeared to be sequence-dependent and were: (i) the largest for Ts in 5'TA3' and 5'TC3' steps, while 2-4 times smaller for 5'-adjacent ones in TT(A,G,C) and TTT(A) runs, (ii) for Cs in 5'TC3' steps 2-4 fold smaller than for adjacent Ts, and (iii) in the presence of Mg2+ generally larger by a sequence-dependent factor: in 5'TC3' steps of about 2 and 4 for Ts and Cs, respectively, in 5'TA3' steps of TTA and TTTA sequences for 3'-terminal Ts of about 3, while for their 5' neighbors o tinctly smaller value of about 2. Comparison of kj data for corresponding Ts located between +1 and -10 regions of Pa promoter in dsDNA and in ssDNA form in the open transcription complex, reported elsewhere, demonstrates that reactivity of pyrimidines in dsDNA is by 2-3 orders of magnitude smaller. The effect of Mg2+ in dsDNA is interpreted in terms of electrostatic barrier to diffusion of MnO4- on DNA surface, which is lowered by diffusive binding of these ions to backbone phosphates, involving also sequence-specific contacts with bases in the minor and major grooves of B-DNA. PMID- 11732621 TI - Repair of gamma-ray-induced base damage in L5178Y sublines is damage type dependent and unrelated to radiation sensitivity. AB - The L5178Y (LY) murine lymphoma sublines LY-R and LY-S are differentially sensitive to ionizing radiation. The high radiation sensitivity of LY-S cells is related to impaired rejoining of DNA double strand breaks. We found previously that the gamma-ray-induced base damage is higher in the more radiosensitive LY-S subline. Here, we examine the role of the repair of ionizing radiation induced base damage in relation to the radiosensitivity difference of these sublines. We used the GS/MS technique to estimate the repair rates of six types of base damage in gamma-irradiated LY cells. All modified DNA bases identified in the course of this study were typical for irradiated chromatin. The total amount of initial base damage was higher in the radiation sensitive LY-S subline than in the radiation resistant LY-R subline. The repair rates of 5-OHMeUra, 5-OHCyt, 8-OHAde were similar in both cell lines, the repair rates of FapyAde and 8-OHGua were higher in the radiosensitive LY-S cell line, whereas the repair of 5-OHUra was faster in its radioresistant counter, the LY-R. Altogether, the repair rates of the y-ray-induced DNA base damage in LY sublines are related neither to the initial amounts of the damaged bases nor to the differential lethal or mutagenic effects of ionizing radiation in these sublines. PMID- 11732622 TI - Background level of 8-oxo-2'-deoxyguanosine in lymphocyte DNA does not correlate with the concentration of antioxidant vitamins in blood plasma. AB - Antioxidant vitamins, being effective free radical scavengers, can protect cellular DNA from oxidative damage. Therefore, in the present study we report on the relationship between basal level of 8-oxo-2'-deoxyguanosine in human lymphocyte DNA and the concentration of antioxidant vitamins (A, C and E). The average level of 8-oxo-2'-deoxyguanosine in lymphocytes of the studied group (15 males and 20 females) was 9.57 per 10(6) dG molecules. The endogenous level of ascorbic acid (vitamin C) in the plasma was, on average, 56.78 microM, while the mean concentrations of retinol (vitamin A) and alpha-tocopherol (vitamin E) were 1.24 uM and 25.74,uM, respectively. No correlations were found between individual 8-oxo-2 micro-deoxyguanosine levels in lymphocyte DNA and endogenous concentration of the vitamins. PMID- 11732623 TI - Lipids and signal transduction in the nucleus. AB - During the last few years a growing amount of data has accumulated showing phospholipid participation in nuclear signal transduction. Very recent data strongly support the hypothesis that signal transduction in the nucleus is autonomic. Local production of inositol polyphosphates, beginning with the activation of phospholipase C is required for their specific function in the nucleus. Enzymes which modify polyphosphoinositols may control gene expression. Much less information is available about the role of other lipids in nuclear signal transduction. The aim of this minireview is to stress what is currently known about nuclear lipids with respect to nuclear signal transduction. PMID- 11732624 TI - Purification and functional reconstitution of intact ral-binding Gtpase activating protein, RLIP76, in artificial liposomes. AB - We have recently shown that RLIP76, a ral-binding GTPase activating protein, mediates ATP-dependent transport of glutathione-conjugates (GS-E) and doxorubicin (DOX) (S. Awasthi et al., Biochemistry 39,9327,2000). Transport function of RLIP76 was found to be intact despite considerable proteolytic fragmentation in preparations used for those studies, suggesting either that the residual intact RLIP76 was responsible for transport activity, or that the transport activity could be reconstituted by fragments of RLIP76. If the former were true, intact RLIP76 would have a much higher specific activity for ATP-hydrolysis than the fragmented protein. We have addressed this question by comparing transport properties of recombinant RLIP76 and human erythrocyte membrane RLIP76 purified in buffers treated with either 100 or 500 microM serine protease inhibitor, PMSF. The purity and identity of recombinant and human erythrocyte RLIP76 was established by SDS/PAGE and Western-blot analysis. These studies confirmed the origin of the 38 kDa protein, previously referred to as DNP-SG ATPase, from RLIP76. Higher PMSF concentration resulted in lower yield of the 38 kDa band and higher yield of intact RLIP76 from both human and recombinant source. In contrast, the substrate-stimulated ATPase activity in presence of DNP-SG, doxorubicin, daunorubicin, or colchicine were unaffected by increased PMSF; similarly, ATP-dependent transport of doxorubicin in proteoliposomes reconstituted with RLIP76 was unaffected by higher PMSF. These results indicated that limited proteolysis by serine proteases does not abrogate the transport function of RLIP76. Comparison of transport kinetics for daunorubicin between recombinant vs human erythrocyte RLIP76 revealed higher specific activity of transport for tissue purified RLIP76, indicating that additional factors present in tissue purified RLIP76 can modulate its transport activity. PMID- 11732625 TI - Effects of wheat germ agglutinin and concanavalin A on the accumulation of glycosaminoglycans in pericellular matrix of human dermal fibroblasts. A comparison with insulin. AB - The effect of insulin, wheat germ agglutinin (WGA), peanut agglutinin (PNA) and concanavalin A (ConA) on [3H]glucosamine incorporation into pericellular glycosaminoglycans (GAGs) was investigated in two lines of cultured human dermal fibroblasts. Insulin and WGA stimulated [3H]glucosamine incorporation into hyaluronic acid (HA) and heparan sulphate (HS) without any alteration of chondroitin sulphate (CS) and dermatan sulphate (DS) contents. ConA increased [3H]glucosamine incorporation into HS, CS and DS, but had no effect on [3Hglucosamine incorporation into HA. PNA affected neither the content, nor the composition of GAGs. In contrast to PNA, ConA and WGA stimulated glycolysis and demonstrated an evident antiproliferative effect on dermal fibroblasts. Thus, both the insulin-like action of WGA and ConA on cultured dermal fibroblasts and the differences between the effects of lectins on modulation of GAGs synthesis appear to be determined by their chemical structure. PMID- 11732626 TI - Kinetics of inactivation of glutamate decarboxylase by cysteine-specific reagents. AB - Mercuric chloride, p-chloromercuribenzoate and 5,5'-dithiobis(2-nitrobenzoic acid) irreversibly inhibited the activity of Escherichia coli glutamate decarboxylase. Their second order rate constants for inactivation are 0.463 microM(-1) min(-1), 0.034 microM(-1) min(-1), 0.018 microM(-1) min(-1), respectively. The characteristics of the inhibition by the three thiol-group reagents supports the idea that cysteinyl residues at the binding sites for the cofactor and/or the substrate are important for enzyme activity in E. coli. PMID- 11732627 TI - Search for polyprenols in leaves of evergreen and deciduous Ericaceae plants. AB - Various species and cultivars of Ericaceae family were checked for the presence of long-chain polyprenols in their leaves. In the genus Rhododendron no polyprenols were found in the ever-green species, while they were present in the deciduous type. The polyprenols were of chain-length of 14-20 isoprene residues and they occurred in the form of acetic acid esters. The polyprenol accumulation is discussed with respect to senescence of leaves. PMID- 11732628 TI - A new view on plant cell. PMID- 11732629 TI - Genomic organization and chromosomal distribution of rat ID elements. AB - Identifier (ID) elements are members of a family of short interspersed nuclear elements (SINEs) in rodents. We investigated the genomic organization and chromosomal distribution of the ID elements in the rat, mouse and Chinese hamster. Southern blot hybridization analysis revealed that the ID elements are widespread in the rat genome, but concentrated in the mouse and Chinese hamster genomes, and that the copy of ID elements in the rat is about 5 times and 50 times that in the mouse and Chinese hamster, respectively. FISH analysis showed that the ID elements are predominantly distributed in the R-band regions of rat chromosomes. In mouse and Chinese hamster chromosomes, no specific distribution pattern of the ID elements was found. Furthermore, we found a distinct group of derivative ID elements in the rat, which contain partially repeated ID core domains, by PCR amplification using an ID core sequence. Such derivatives were not found in either the mouse or Chinese hamster. These findings suggest that explosive amplification of the ID elements in the rat has been accompanied by the occurrence of derivative ID elements and a predominant localization to the R-band regions. Similar associations found in the Alu family, one of the human SINEs, allow us to speculate that the rat ID elements and the human Alu family have analogous functions in chromosomal organization. PMID- 11732630 TI - Mating system and genetic variance in a polygynous mustelid, the European polecat. AB - The population genetic implications of mating system were investigated in European polecat Mustela putorius populations from western France, combining radiotracking survey and allozyme variation analysis. Mating period occurred from February to June and polecats showed a strategy of successive polygyny, a male consorting with 1.44 females during a brief period (2.9 days). Relatedness was largely sex biased, females (21%) being almost twice more related than males (13%) suggesting a natal philopatry. Nonetheless, breeding dispersal pattern appeared relatively complex. Males were the sex dispersing but the main strategy for male polecats consisted of short-term mating excursions in adjacent females ranges whereas long-distance dispersal only constituted an alternative breeding strategy. Despite their allozymic polymorphism level reaching 24% at p<0.05 for 38 scored loci, populations showed a high heterozygote deficiency as revealed by the F(IS) index averaging F(IS) = 0.383. Thus the mating system of such solitary mustelids may be poorly efficient to prevent inbreeding within populations. PMID- 11732631 TI - Molecular phylogeny of butterflies Parnassius glacialis and P. stubbendorfii at various localities in East Asia. AB - The phylogeny of butterflies, Parnassius stubbendorfii and P. glacialis, collected at various localities in the Japan archipelago and the eastern part of the Asian continent was analyzed using mitochondrial DNA sequences coding for NADH dehydrogenase subunit 5 (805 bp). The molecular phylogenetic trees revealed that P. glacialis and P. stubbendorfii diverged from a common ancestor, and then the populations inhabiting the Japan archipelago and the Asian continent diverged in each species. The reliability of these divergences was supported by high bootstrap values. The divergences within the Japan archipelago and within the Asian continent in each species were unclear because of low bootstrap values. The genetic distance and a rough time-estimation in the UPGMA tree suggest that the both populations of P. glacialis and P. stubbendorfii may have been isolated in the Japan archipelago at the early time (about 1.7-2.0 Mya) of the glacial period in the Pleistocene. The genetic distance between the Japanese and the continental subspecies may be large enough that they can be classified as different species, in comparison with the genetic distances among some other parnassian species. PMID- 11732632 TI - Karyotypes and Giemsa C-banding patterns of Zebrina pendula, Z. purpusii and Setcreasea purpurea, compared with those of Tradescantia ohiensis. AB - It has been proposed that the genera Zebrina and Setcreasea of the family Commelinaceae should be united and reunited, respectively, with the genus Tradescantia, mainly based on morphological studies. In the present study, karyotypes and Giemsa C-banding patterns in the root-tip cells of three Zebrina and two Setcreasea clones were analyzed, and were compared with those of a triploid Tradescantia clone. Z. pendula and Z. purpusii (both 2n = 24) were found to have similar karyotypes (4 M + 6 ST + 14 T; M = meta-, ST = subtelo-, T = telocentric chromosomes), while Z. pendula cv Quadricolor (2n = 23) had a unique karyotype (6 M + 5 ST + 11 T + 1 SA; SA = short acrocentric chromosome). The only clear difference between Z. pendula and Z. purpusii was that one and two subtelocentric chromosomes, respectively, had satellites at the short arms. Two clones of S. purpurea (2n = 24) had karyotypes (8 M + 8 M' + 8 SM; M' = nearly meta-, SM = submetacentric chromosomes) similar to each other. T. ohiensis (2n = 18) had a symmetric karyotype (9 M + 9 SM) consisting of larger chromosomes than S. purpurea. Many clear Giemsa C-bands were detected, in addition to centromeric bands in all chromosomes of all clones. Z. pendula and Z. purpusii commonly had single clear interstitial bands in eight telocentric chromosomes each, but they also had unique telomeric and other interstitial bands, respectively. Z. pendula cv Quadricolor had a unique banding pattern, i.e., satellite bands in the unique short chromosome, telomeric bands at the long arms of all metacentric chromosomes, and single interstitial bands in six telocentric chromosomes. Two clones of S. purpurea had telomeric bands at many chromosome arms and satellite bands in two nearly metacentric and one submetacentric chromosomes, but some differences were found between them. On the other hand, all the chromosomes of T. ohiensis had telomeric bands at both arms, and three submetacentric chromosomes had satellite bands. These result prove structural differentiation of chromosomes occurred among the clones, especially in Zebrina, and show that S. purpurea is relatively close to T. ohiensis, while Zebrina is obviously distant from the other two genera. Therefore, there remains a question cytologically at least for uniting Zebrina with Tradescantia. PMID- 11732633 TI - Suitability of AFLP markers for the study of genetic relationships among Korean native dogs. AB - To determine the genetic relationships among domestic dog breeds, we performed both a sequence comparison of mitochondrial DNA (mtDNA) and an amplified fragment length polymorphisms (AFLP) analysis. Three of four regions of mtDNA, cytochrome b, cytochrome oxidase subunit II, and 16S rRNA genes were highly homogeneous among dog breeds, whereas the other region, the control region, showed relatively high polymorphisms with a maximum percentage difference of 3.18%. However, the control region showed extensive polymorphism even within breeds, and the relationship tree derived from the data could not clearly delimit distinct breeds. 19 EcoRI/MseI primer combinations were used to generate AFLP markers among 25 dogs from 11 breeds including three Korean native dogs. These amplification reactions allowed the detection of more than 1900 amplification products of which 408 were identified as polymorphic bands. Unrooted neighbor joining tree based on dissimilarity values showed that the Korean native dogs were clustered together with the Asian dogs and that the Asian originated dogs were clustered separately from Western originated dogs. A consensus tree using parsimony method also showed Korean native dogs were grouped separately from the other dogs with moderate bootstrap values. Taken together, it is concluded that AFLP analysis is a more informative tool for revealing genetic relationships among dog breeds than mtDNA sequence comparison. PMID- 11732634 TI - BAC FISH analysis in Allium cepa. AB - Onion (Allium cepa L.; 1C=15,000 Mb) is an agriculturally important plant. The genome of onion has been extensively studied at the conventional cytogenetic level, but molecular analyses have lagged behind due to its large genome size. To overcome this bottleneck, a partial bacterial artificial chromosome (BAC) library of onion was constructed. The average insert size of the BAC library was about 100 kb. A total of 48,000 clones, corresponding to 0.32 genome equivalent, were obtained. Fluorescent in situ hybridization (FISH) screening resulted in identification of BAC clones localized on centromeric, telomeric, or several limited interstitial chromosomal regions, although most of the clones hybridized with entire chromosomes. The partial BAC library proved to be a useful resource for molecular cytogenetic studies of onion, and should be useful for further mapping and sequencing studies of important genes of this plant. BAC FISH screening is a powerful method for identification of molecular cytogenetic markers in large-genome plants. PMID- 11732635 TI - Characterization of a fission yeast mutant which displays defects in cell wall integrity and cytokinesis. AB - The fission yeast cps6-153 mutant was originally isolated based on its hypersensitivity to the spindle poison isopropyl N-3-chlorophenyl carbamate (CIPC). The mutant also shows defects in both cell wall integrity and cytokinesis, resulting in the accumulation of unseparated cells with weakened cell walls. The arrested cells display a disoriented alignment of cytoplasmic microtubules. When the mutant cells are cultivated at high temperature (35 degrees C), both cell walls and septa become very thick. Electron microscopy revealed the disorganized structure of the thickened cell walls and septa, in which fibrillar components were not completely masked with an amorphous matrix. rad25+ was cloned from a genomic library by complementation of the mutant phenotypes, suggesting the involvement of Rad25p, one of two 14-3-3 proteins in S. pombe, in the pathway of cell wall integrity and cytokinesis. PMID- 11732636 TI - Working memory in ADHD: prefrontal/parietal connections. AB - Current theories of dopaminergic and noradrenergic mechanisms, which are thought to be of importance in the regulation of attention are reviewed. A biphasic model of dopaminergic function is described, in which tonic dopamine exerts a suppressive influence on subcortical dopamine systems by altering tonic/phasic dopaminergic relationships. Noradrenergic mechanisms are of importance in modulating sensory processing at the prefrontal cortical level. The work of Silberstein and colleagues utilizing Steady-State Visually Evoked Potential, during the course of an A-X Continuous Performance Task enables examination of the spatial distribution and dynamics of electrical brain activity during the task. The maintenance of activation in the interval between A and X provides a measure of working memory, thought to be related to prefrontal-parietal activation, which is facilitated by administration of methylphenidate to children with ADHD, suggesting that working memory may be a core deficiency in children with ADHD. While tonic dopamine activity in ventral striatum/accumbens gates inhibitory activity, dorsolateral prefrontal-parietal connections allow maintenance of working memory required for goal completion. PMID- 11732637 TI - Pharmacology of appetite suppression: implication for the treatment of obesity. AB - Given the current global epidemic of obesity there is a demand for new anti obesity drugs to overcome the problem. Many pharmacological agents reduce food intake and significantly decrease body mass when administered to animals but affect feeding behaviour in a profoundly different way indicating the variety of biological mechanisms by which such agents act (appetite verses non-appetite). More limited clinical data demonstrates that some of the same drugs produce decreases in food intake and weight loss in humans. A few of these drugs do so by modifying the functioning of the appetite system as measured by subjective changes in feelings of hunger and fullness (indices of satiety). These drugs that modify the daily flux of appetite could be considered as 'appetite suppressants' with clinical potential as anti-obesity agents. Drugs that can be considered suitable candidates for appetite suppressants are agents that enhance peripherally satiety peptide systems (such as CCK, Bombesin/GRP, Enterostatin and GLP-1), alter the CNS levels of various hypothalamic neuropeptides (NPY, Galanin, Orexin, CART and Melanocortins) or monoamine neurotransmitters (such as serotonin, nor-adrenaline and possibly dopamine). Recently, the hormone leptin has become regarded as a key hormonal signal linking adipose tissue status with a number of key central nervous system circuits (NPY, CART, CRF, Melanocortins and possibly Orexins). This tonic system may also provide drug targets for the control of appetite. Any changes induced by a potential appetite suppressant should be considered in terms of the (i) psychological experience and behavioural expression of appetite, (ii) metabolism and peripheral physiology, and (iii) functioning of CNS neural pathways. In humans, such modulation of appetite will involve changes in total caloric consumption, subjective changes in feelings of hunger and fullness, preferences for specific food items, and general macronutrient preferences. These may be expressed behaviourally as changes in meal patterns, snacking behaviour and food choice. Within the next 20 years it is certain that clinicians will have a new range of anti-obesity compounds available to choose from. Such novel compounds may act on a single component of the appetite system or target a combination of these components detailed in this review. Such compounds used in combination with life style changes and dietary intervention may be critical in dealing with the rising world epidemic of obesity. PMID- 11732638 TI - Gene therapy: optimising DNA delivery to the nucleus. AB - Gene therapy, the expression in cells of genetic material that has therapeutic activity, holds great promise for the treatment of a number of human diseases. A gene delivery vehicle, or vector, that may be of viral or non-viral origin, is generally used to carry the genetic material. Viral vectors have been developed that exclude immunogenic genes while taking advantage of the genes responsible for proficient integration of the viral genome into that of the host. In this way, viral vectors improve the probability of long-term expression of the therapeutic gene, whereas non-viral vectors, that are not as efficient at introducing and maintaining foreign gene expression, have the advantage of being non-pathogenic and non-immunogenic. Although thousands of patients have been involved in clinical trials for gene therapy, using hundreds of different protocols, true success has been limited. A major limitation of gene therapy approaches, especially when non-viral vectors are used, is the poor efficiency of DNA delivery to the nucleus; a crucial step to ensure ultimate expression of the therapeutic gene product. Here we review existing gene delivery approaches and, in particular, explore the possibility of enhancing non-viral gene delivery to the nucleus by incorporating specific nuclear targeting sequences in vectors, using a range of different strategies. PMID- 11732639 TI - The Ca2+-activated K+ channel of intermediate conductance: a molecular target for novel treatments? AB - This review discusses the Ca2+-activated K+ channels of intermediate conductance (IK channels), and their historical discovery in erythrocytes, their classical biophysical characteristics, physiological function, molecular biology as well as their role as possible molecular targets for pharmacological intervention in various diseases. The first described Ca2+-activated K+ channel ever - the so called Gard6s channel from human erythrocytes--is an IK channel. The "I" denominates the intermediate conductance that distinguishes the IK channels from the related Ca2+-activated K+ channels of small (SK) or large (BK) conductance. The recent cloning of the human IK channel gene (KCNN4) enabled a detailed mapping of the expression in various tissues. IK channel expression is found predominantly in cells of the blood, in epithelia and endothelia. An important physiological role of IK channels is to set the membrane potential at fairly negative values and thereby to build up large electrical gradients for the passive transport of ions such as Cl- efflux driving water and Na+ secretion from epithelia, and Ca2+ influx controlling T-lymphocyte proliferation. The molecular cloning of IK and SK channels has revealed that both channels gain their Ca2+ sensitivity from tightly bound calmodulin (CaM). The IK channel is potently blocked by the scorpion toxin charybdotoxin (ChTx) and the antimycotic clotrimazole (CLT). CLT has been in clinical trials for the treatment of sickle cell disease, diarrhea and ameliorates the symptoms of rheumatoid arthritis. However, inhibition of cytochrome P450 enzymes by CLT limits its therapeutic value, but new drug candidates are entering the stage. It is discussed whether pharmacological modulation of IK channels may be beneficial in sickle cell anemia, cystic fibrosis, secretory diarrhea, craft-versus-host disease and autoimmune diseases. PMID- 11732640 TI - Raloxifene. AB - Efforts to interfere with the initiation and promotion of breast and other cancers by endocrine manipulation are not new. It is of obvious benefit to cancer patients to administer substances that combine minimal general toxicity with maximal oestrogen inhibition. Raloxifene is a relatively recent addition to a group of compounds loosely designated as antioestrogens, which implies their ability to antagonize oestrogen effects via competitive binding to the various receptors. This is a reductionist simplification, since their effect varies and ranges from interaction with lipid transduction cascades, covalent binding to proteins and DNA, regulation of growth factors, erbB2, mdr1 and probably p53 expression, complexing with E-cadherin/catenin to active induction of apoptosis and many other effects on the genome. Also, the action of most antioestrogens is not solely antagonistic and different compounds do exert some agonistic effects in various tissues. Apart from some "pure" antioestrogens, the benzothiophene derivative Raloxifene has been found to combine a high degree of selective oestrogen suppression with several other desirable characteristics, such as reduction of bone demineralisation and antiatherogenic effects without endometrial stimulation. It is well tolerated, has been successfully tested as a chemopreventive agent for breast cancer in certain groups of the population and does not prevent ovulation in women with normal menstrual cycles. Certainly, Raloxifene is only another forerunner of upcoming "designer" oestrogen modulators, but it represents a welcome addition to the therapeutic choices available for the control of some menopausal problems as well as for the prevention and treatment of breast cancer, as outlined in the following brief review. PMID- 11732641 TI - An approach for the rational design of new antituberculosis agents. AB - Tuberculosis (TB) kills more youth and adults than any other infectious disease in the world today. The emergence of new strains of Mycobacterium tuberculosis resistant to some or all current antituberculosis drugs is a serious and crescent problem. The resistance is often a corollary to HIV infection and drug-resistant TB is more difficult and more expensive to treat, besides to be more likely fatal. Thus, it is still necessary to search for new antimycobacterial agents. The identification of novel targets need the identification of biochemical pathways specific to mycobacteria and related organisms. Many unique metabolic processes occur during the biosynthesis of mycobacterial cell wall components. In this report, we examine one of these attractive targets for the rational design of new antituberculosis agents--the mycolic acids. PMID- 11732642 TI - Which surrogate marker can be used to assess the effectiveness of the laboratory and its contribution to clinical outcome? AB - Assessment of the effectiveness of the clinical laboratory and its contribution to outcomes is gaining increasing emphasis, as part of the overall attempts at making clinical services more transparent and accountable. Tools traditionally used in the assessment of laboratory effectiveness and efficiency have included laboratory accreditation, Q-probes, performance in quality assurance programmes and staffing and cost issues. There is, however, a need to introduce different measures that highlight the laboratory efficiency and contribution to clinical effectiveness and outcomes. Such measures should, ideally, be quantifiable and evidence-based. The use of markers of efficiency and effectiveness could be used as tools to aid this process. Such markers could include incident reporting, the appropriateness of assay repertoire, adding value to reports, the quality of comments made, provision of information on the effect of analytical and biological variation on results, cascading requests to help making diagnoses and unearthing such diagnoses. We suggest that these measures contribute towards the implementation of the clinical governance agenda in relation to the laboratory, and could be used as indicators in laboratory accreditation. PMID- 11732643 TI - Endocrinology of gynaecomastia. AB - Gynaecomastia is the most common disorder of the male breast. It can occur at any age, and for this reason laboratory investigations may be requested by clinicians from many specialties. Gynaecomastia may occur transiently in neonates. It may also occur transiently during puberty, when it is common and generally benign. It must, however, be regarded as unusual in prepubertal children and all young and middle-aged men. Although iatrogenic and benign gynaecomastia are common in the elderly, further investigations may still be justified since breast cancer or other neoplasm must be ruled out. Biochemical investigations, when warranted, are aimed at establishing an underlying cause. Endocrine investigations might include serum oestradiol (or oestrone if available), testosterone, luteinizing hormone, sex-hormone-binding globulin, human chorionic gonadotrophin, prolactin and thyroid function tests. In this review, the source and role of oestrogens in men, the androgen oestrogen dynamics, the causes and clinical entities of gynaecomastia, and interpretation of laboratory tests are described. PMID- 11732644 TI - Bisphosphonates: an overview with special reference to alendronate. AB - Bisphosphonates, analogues of pyrophosphate, are potent inhibitors of osteoclast mediated bone resorption. They are used in the treatment of Paget's disease of bone, hypercalcaemia and osteolytic bone disease of malignancy, primary and secondary hyperparathyroidism, and in osteoporosis. Bisphosphonate treatment causes an early reduction in bone resorption followed by a later reduction in bone formation. The early inhibition of bone resorption induces a reduction in serum calcium which leads to increased parathyroid hormone (PTH), and subsequently an increase in 1,25-dihydroxyvitamin D. The secondary hyperparathyroidism of bisphosphonate treatment also leads to urinary calcium conservation and phosphaturia, and a reduction in serum phosphate. The increase in the PTH following bisphosphonate therapy is a response to the change in serum calcium and can occur even when there is hypercalcaemia, and this can cause confusion in the interpretation of PTH results. The hypocalcaemic response to bisphosphonates is occasionally severe, especially in patients with hypoparathyroidism. The recent elucidation of bisphosphonate action at the cellular level on the mevalonate pathway has led to interest in its effects on lipoprotein metabolism, which may prove to be of clinical significance. Newer and more potent bisphosphonates are currently undergoing clinical trials in malignant bone disease and osteoporosis, and will lead to further advances in the optimal management of these conditions. PMID- 11732645 TI - Assessment of homocysteine as a cardiovascular risk factor in clinical practice. AB - Elevated plasma total homocysteine concentrations are a marker of vitamin deficiency and a risk factor for cardiovascular disease. It is possible that vitamin supplementation with folic acid and other B vitamins, which lower plasma homocysteine concentrations, may reduce the risk of cardiovascular disease. Large scale clinical trials are currently underway to assess the homocysteine hypothesis of cardiovascular disease. Pending the outcome of such trials, measurement of plasma homocysteine concentrations in people at high risk of cardiovascular disease may help to identify patients who could benefit from more intensive treatment of classical cardiovascular risk factors. The introduction of immunoassays for homocysteine determination has made assessment of homocysteine status accessible to most routine hospital laboratories, and this review summarizes the evidence on why and how to assess homocysteine as a risk factor for cardiovascular disease in clinical practice. PMID- 11732646 TI - Prostate specific antigen: biology, biochemistry and available commercial assays. AB - Prostate specific antigen (PSA) is the marker of choice in the management of prostate cancer. However, PSA is not a simple molecule, existing in the serum in five isoforms and a number of molecular configurations and complexes. The elucidation of the biochemistry of PSA has increased the potential use of the marker in the diagnosis of prostate malignancy. This review summarizes the clinical use of PSA in the management of prostate disease and the assays available in the UK. Assay calibration in relation to the World Health Organization 1st International Standard for Prostate Specific Antigen (90:10) has increased conformity between the various commercial assay kits, and the non equimolar kits have largely been superseded or withdrawn. Special reference is made to evaluations performed on behalf of the Medical Devices Agency of the Department of Health. PMID- 11732647 TI - Biochemical detection and monitoring of alcohol abuse and abstinence. AB - The merits and limitations of traditional and new markers for alcohol abuse (and abstinence) are critically examined for detection and monitoring of alcoholics, hazardous drinkers and binge drinkers. The traditional markers discussed include gamma-glutamyltransferase (GGT), aspartate and alanine aminotransaminases (AST, ALT) and mean corpuscular volume (MCV); new markers include mitochondrial AST, carbohydrate-deficient transferrin (CDT), serum/urine 5-hydroxytryptophol, beta hexosaminidase and acetaldehyde adducts. The strengths and weaknesses of several of the self-reporting screening questionnaires are also explored. No laboratory test is reliable enough on its own to support a diagnosis of alcoholism. Sensitivities and specificities vary considerably and depend on the population concerned. GGT continues to remain the test that combines greatest convenience and sensitivity: its diagnostic accuracy can be enhanced by combination with other traditional markers (AST, ALT, MCV). None of the newer markers offers significant advantage, although CDT seems to be better at monitoring patients for increased alcohol consumption or progress towards abstinence. PMID- 11732648 TI - Determination of plasma free carnitine by electrospray tandem mass spectrometry. AB - We have evaluated a tandem mass spectrometry method for determining free carnitine concentrations in plasma and have compared its performance with that of an existing radioenzymatic assay. In this method, plasma was mixed with an internal standard, carnitine-d3 (600 nmol/L), and butylated before analysis on a Quattro II tandem mass spectrometer. The detection limit of the tandem mass spectrometric (MS) method was 4 micromol/L and carryover between samples was 2.2%. Precision of the method was 1.6-5.4% between injections and 4.0-5.6% between samples. Between-batch precision was 10-13%. The method was linear up to a free carnitine concentration of 300 micromol/L and good agreement was found with the existing radioenzymatic assay. We conclude that the tandem MS method is a precise and robust method for the determination of free carnitine concentrations in plasma that overcomes the disadvantages of the radiochemical method. PMID- 11732649 TI - Useful tests on pleural fluid that distinguish transudates from exudates. AB - We aimed to compare the classic Light's criteria with different testing strategies in an effort to improve the accuracy of pleural fluid (PF) categorization. Thirty-two patients with transudates and 140 with exudates on the basis of their clinical diagnosis were entered into the study. We examined the discriminative properties of 10 analytes in the identification of PF, both singly and in combination with an 'or' rule, to see which was best in distinguishing a transudate from an exudate. A combination of PF lactate dehydrogenase (LD) > 307 U/L (two-thirds of the upper limit of the serum LD reference range) with either PF cholesterol > 1.55 mmol/L or PF to serum protein ratio > 0.5 had a diagnostic accuracy similar to that of Light's criteria. We suggest the use of PF LD and cholesterol in combination as an alternative method for distinguishing pleural transudates from exudates. This test combination avoids the need for venepuncture and the simultaneous collection of a blood sample. PMID- 11732650 TI - Effect of ambient temperature on analytical and clinical performance of a blood glucose monitoring system: Omnitest Sensor glucose meter. AB - The effect of ambient temperature on the analytical and clinical performance of a glucose meter was examined. A total of 114 venous whole blood samples were analysed for glucose by a reference method, and by a glucose meter at 21-22 degrees C, room temperatures, 26-27 degrees C and 33-34 degrees C. Glucose meter readings at each temperature were compared with the reference values and evaluated by analysis of variance, Spearman's correlation, the percentage of glucose meter readings within +/- 10% of the reference value and error grid analysis. Analysis of covariance was used to determine the effect of temperature on glucose meter readings. There were no significant differences in the glucose meter readings and in accuracy of the meter readings between different temperatures. Temperature was not a significant independent determinant of the glucose meter readings. For each glucose concentration, the precision of the meter and clinical performance were comparable between the different temperatures. In conclusion, ambient temperature does not affect the accuracy, precision and clinical performance of the Omnitest Sensor. PMID- 11732651 TI - Performance of a novel spectrophotometric lithium assay on a routine biochemistry analyser. AB - A new spectrophotometric lithium assay was assessed after adaption to a Hitachi 917. Results on this analyser were in close agreement with lithium measured on a flame photometer (95% confidence interval+/-0.06 mmol/L, n = 120). Assay precision was <2.5% over a range of lithium concentrations from 0.5-3.0 mmol/L. The assay was linear to 3 mmol/L and interference from haemolysis, icterus, lipaemia and other metal ions was minimal. The assay is a useful substitute or "back-up" for traditional lithium assays. Improved "on-board" calibration stability (currently 24 h) is required by improving the stability of uncapped reagent. PMID- 11732652 TI - In vitro cold activation of complement shown by an overestimation of total complement 4: a study in patients with hepatitis C virus infection. AB - Total complement 4 (C4) levels, when analysed on the Beckman Array nephelometer, were found to increase number of serum specimens [predominantly from patients with hepatitis C virus (HCV) infection] after overnight storage at 4 degrees C. In order to investigate whether the phenomenon of in vitro cold-dependent activation of complement (CDAC) was the explanation for this increase, paired specimens were collected from 63 patients with HCV infection in tubes with no anticoagulant (serum) and in tubes containing EDTA (which inhibits complement activation). C4 levels increased after overnight storage at 4 degrees C in 33 serum specimens (52%). In contrast, no increase in C4 levels was observed in any of the 63 EDTA specimens. Immunofixation of intact and activated C4 products confirmed that complement activation had taken place in the serum specimens in which C4 levels had increased after storage. There was a higher frequency of hepatitis C viraemia (P<0.0001), HCV antibody positivity (P<0.05) and the presence of rheumatoid factor (P<0.05) in the group of patients in whose serum samples CDAC had occurred (n = 33) than in the other group (n = 30). As a result of our findings on C4 analysis in stored serum specimens, we would recommend potassium EDTA plasma as the specimen of choice for complement analysis on the Beckman Array. PMID- 11732654 TI - Determination of D-sorbitol in human erythrocytes by an enzymatic fluorometric method with an improved deproteinization procedure. AB - We developed an improved enzymatic assay of D-sorbitol in human erythrocytes by employing highly specific D-sorbitol dehydrogenase from Pseudomonas sp. (EC 1.1.1.14) and replacing perchloric acid (HClO4) and potassium carbonate (K2CO3), generally used for deproteinization, with sodium hydroxide (NaOH) and zinc sulphate (ZnSO4). In this assay, erythrocytes were separated from plasma by centrifugation and washed once with physiological saline. Subsequently, the erythrocytes were lysed with distilled water and proteins precipitated with NaOH and ZnSO4. After centrifugation, the resulting colourless supernatant was mixed with a glycine buffer (pH 9.0) containing NAD+ and D-sorbitol dehydrogenase. After incubation for 30 min at 37 degrees C, the NADH produced was measured fluorimetrically. The fluorescence intensities were corrected for sample blanks, and the values of D-sorbitol were normalized for haemoglobin content. The method had an analytical range of 1-180 micromol/L. The intra- and inter-assay precisions were < 3.3% and < 5.8%, respectively. The detection limit was 0.65 micromol/L. In terms of the linearity, precision and sensitivity, the improved method using NaOH and ZnSO4 was superior to the conventional method using HClO4 and K2CO3. PMID- 11732653 TI - Protection by metal complexes with SOD-mimetic activity against oxidative gastric injury induced by indomethacin and ethanol in rats. AB - We have investigated the protective effect of oral administration of copper and manganese complexes with superoxide dismutase (SOD)-mimetic activity against oxidative gastric mucosal injury induced by the non-steroidal anti-inflammatory drug indometacin with ethanol in the rat. The total area of the gastric lesions and lipid peroxidation were significantly increased 1 h after oral administration of indometacin (15 mg/kg) and ethanol, indicating an acute oxidative injury. The activities of SOD, catalase (CAT), glutathione-S-transferase (GST) and glutathione content were significantly decreased in the gastric mucosa by indometacin plus ethanol. Manganese or copper complexes showed SOD-mimetic activity. Pretreatment with these complexes protected against gastric mucosal lesions and decreased lipid peroxides, as well as attenuating the decrease in the activities of SOD, CAT and GST in gastric mucosa. These findings suggest that active oxygen species and lipid peroxidation play an important role in the pathogenesis of gastric mucosal injury induced by indometacin. In addition, we have shown that Mn and Cu complexes have gastroprotective properties against ulceration induced by indometacin plus ethanol. The present results suggest that appropriate copper or manganese complex supplementation may potentially provide prophylaxis or therapy for some pathologies associated with excessive free radical production and inhibited SOD activity. PMID- 11732655 TI - Monoclonal IgM: difficulties with correct measurement. AB - We describe three monoclonal IgM paraproteins for which nephelometric IgM quantification generated inaccurate results. PMID- 11732656 TI - Limitation of the Vitros dry-slide technique in measuring high concentrations of salicylate in plasma. AB - The linearity of the Vitros dry-slide method for plasma salicylate was assessed in two ways: serial concentrations of sodium salicylate were added to fresh lithium heparin plasma, and the salicylate was determined both neat and in dilution. Vitros salicylate results submitted to the Heathcontrol External Quality Assessment Scheme were compared to the spike value. Similar loss of linearity was observed in both cases. Serious salicylate overdosage requiring specific clinical treatment may have been underestimated. PMID- 11732657 TI - Appropriate design of biochemistry request cards can promote the use of protocols and reduce unnecessary investigations. AB - Attempts to control laboratory workload in the past always proved to be short lived. We designed a request card based on clinical conditions for a gynaecology out-patient clinic to help clinical staff to adhere to agreed investigation protocols. Each request card had a tick-box associated with a clinical condition. The tests performed for each condition were advertised on a poster in the clinic. The workload was assessed for a 2-year period before and after the introduction of these cards. The numbers of all endocrine tests, except progesterone and sex hormone binding globulin, fell by 19% (P<0.0001) whereas the clinical cases seen remained steady at 227 per month. This report demonstrates that agreed protocols can be implemented but that a persistent aide-memoire is necessary. PMID- 11732658 TI - Limitation of the Vitros dry-slide technique in measuring high concentrations of salicylate in plasma. PMID- 11732659 TI - Tomatoes and blood: changing dietary habits and faecal occult blood testing. PMID- 11732661 TI - New directions in the treatment and prevention of deep vein thrombosis. PMID- 11732660 TI - Evaluation of specific high-performance liquid-chromatographic determinations of urinary metanephrine and normetanephrine by comparison with isotope dilution mass spectrometry. AB - A method for the determination of metanephrine (MN; also known as metadrenaline), normetanephrine (NMN; also known as normetadrenaline) and 3-methoxytyramine (3 MT) in human urine using high-performance liquid chromatography followed by electrochemical detection (ECD) was validated primarily by comparing the results with those obtained by a gas chromatography mass spectrometry (GC-MS) reference method. Correlation coefficients of 0.93, 0.94 and 0.91 were obtained for MN, NMN and 3-MT, respectively, in a group of healthy controls consisting of 30 women and 30 men. A systematic difference was detected only for 3-MT (-16%). Further tests of accuracy (linearity and recovery) and precision demonstrated that the described method must be considered to be a reliable approach to assess urinary metanephrines in the diagnosis of phaeochromocytoma. At lower concentrations (MN, 248 nmol/L; NMN, 434 nmol/L; 3-MT, 402 nmol/L), within-assay coefficients of variation were close to 5% or less (5.3, 4.6 and 2.2%, respectively) and between assay coefficients of variation were 8.9, 11.2 and 12.3%, respectively, for the same low levels. This raises the possibility that this method can also be applied to assess urinary free, unconjugated metanephrines. Sex differences were detected for MN and NMN excretion when expressed in nmol per 24h and nmol/mmol creatinine, respectively, by both ECD and GC-MS methods. PMID- 11732662 TI - Treatment of deep vein thrombosis using low-molecular-weight heparins. AB - Because it eliminates the need for intravenous infusions and lengthy hospitalizations, use of low-molecular-weight heparins (LMWHs) for the treatment of deep vein thrombosis (DVT) can improve patient outcomes of safety and efficacy and reduce the cost of medical care. In a meta-analysis of randomized controlled trials involving more than 3,500 patients, treatment with LMWH versus unfractionated heparin was associated with lower mortality, a lower incidence of major bleeding episodes and thrombocytopenia, and an overall reduction of about 30% in morbidity from recurrent thromboembolism and bleeding. The meta-analysis also demonstrated cost savings of $1,900 per patient for those treated with LMWH. More recent studies evaluating several LMWHs in both hospital and outpatient settings have shown that the drugs are more effective in reducing risk for recurrent events and complications than unfractionated heparin. The use of LMWHs for outpatient treatment of DVT has, of necessity, given rise to specific protocols, inclusion and exclusion criteria, follow-up procedures, and multidisciplinary approaches to outpatient management. These guidelines are reviewed here, as are special considerations for patients who are obese or who have renal insufficiency. PMID- 11732663 TI - Prophylaxis strategies for patients with acute venous thromboembolism. AB - Pulmonary embolism is one of the most common causes of unexpected death in hospitalized patients and one of the top diseases leading to medical malpractice lawsuits. In order to effectively prevent venous thromboembolism (VTE), physicians must assess patients' risk factors and stratify their risk accordingly. Studies show general medical patients are most likely to suffer from deep vein thrombosis. Research also indicates that once-daily prophylaxis of such patients with 40 mg of the low-molecular-weight heparin (LMWH) enoxaparin is at least as effective or more effective as prophylaxis with unfractionated heparin, and may be preferable in some populations. It is now being recommended that hospitals develop formal strategies that address the prevention of thromboembolic complications and that general medical patients at risk of VTE receive unfractionated heparin or LMWH. PMID- 11732664 TI - Economic issues in the treatment and prevention of deep vein thrombosis from a managed care perspective. AB - Treatment and prevention of deep vein thrombosis (DVT) raise a number of important economic issues. Economic burden of the disease is a primary concern. Enrollment trends indicate that the patient population mix in managed care health plans is changing. Health plans are enrolling a growing number of members who are at higher risk for DVT. Accordingly, managed care health plans will increasingly need to focus attention on the development of high-quality and cost-effective strategies for treatment and prevention of DVT. It is widely agreed that the use of low-molecular-weight heparin (LMWH) is safe and effective, and there is also good evidence that its use is cost effective in selected indications. The shift from inpatient to outpatient treatment that is made possible by substitution of LMWH for conventional unfractionated heparin brings with it a need to restructure healthcare delivery resources. This may have substantial economic impact on both providers and health plan members. Growing recognition of the DVT risk associated with various disease states, as well as the costly long-term sequelae of DVT, is spurring a trend to broader use of LMWH for prophylaxis. The relative cost effectiveness of prevention depends on the tradeoff between an expenditure that is made today and an uncertain future benefit. What is needed now is better evidence as a basis for decisions regarding DVT prophylaxis and clinical studies that track the long-term clinical and economic impact of DVT. PMID- 11732665 TI - The use of pharmacoeconomics in formulary development: can this improve the way deep vein thrombosis is treated? AB - A study of the Prescription Solutions database revealed that in 2000 the average pharmacy plus medical costs per patient with deep vein thrombosis (DVT) were $60,019. The total charges for this group of patients in this organization were $389 million. The cost of treating these patients is relatively high, and there may be opportunities to significantly improve the quality and overall cost effectiveness of their care. One example of how this might be accomplished is by developing a pharmacy-based DVT predictive model that could be used proactively to identify patients who may be at risk for DVT and who might benefit from early intervention with low-molecular-weight heparin therapy prophylaxis. PMID- 11732666 TI - The use of a protocol for the treatment of deep vein thrombosis. PMID- 11732667 TI - Nutritive and economic values of high oil corn in layer diet. AB - Two layer feeding trials were conducted to demonstrate the nutritive and economic values of recently developed high oil corn (HOC) in Korea. A corn-soybean meal based commercial layer diet was chosen as the control diet. The yellow dent corn in the control diet was replaced with HOC to give an isocaloric diet, or replaced with HOC on a 1:1 basis to give a high energy diet. In Trial 1, 510 23-wk-old ISA Brown layers were allotted to three dietary treatments with five replicates per treatment. In Trial 2, 600 38-wk-old Hy-Line Brown layers were allotted to three dietary treatments, again with five replicates per treatment. Both trials were conducted for 15 wk. To measure the ME values of typical corn and HOC, two metabolism trials were performed with layers and adult roosters. The HOC used in this trial contained approximately 94% higher crude fat (6.60% as-fed basis) compared with typical corns. The gross energy, AMEn, and TME values of HOC, are 5.7 to 7.7% higher than those of typical corns, indicating that the energy use of each corn were similar. Oil from the HOC contains 6.5 to 8.3% more oleic acid and 6 to 7% less linoleic acid than oil from typical corns. HOC feeding, on an isocaloric basis or on 1:1 replacement with typical corn, did not exert any effect on various laying performances, including the physical quality of egg. This result reflects the quality of the commercial diet chosen as the control diet, which was already fairly good, such that the performance was already maximal. The polyunsaturated fatty acid content in yolk from hens fed HOC was higher than that from hens fed typical corns, reflecting higher linoleic acid content in the HOC. HOC feeding decreased the saturated fatty acid content in the yolk, due primarily to decreased palmitic acid. If used alone replacing typical corn completely in a layer diet, the acceptance price of HOC was estimated to be 154 won/kg when the price of typical corn was 131 won/kg (118:100). When both corns were allowed to be used, the acceptance price of HOC increased to 184 won/kg (140:100), indicating that a lot cheaper layer diet can be formulated when both HOC and typical corn are used in laying hen diet formulation. PMID- 11732668 TI - Growth and immunity of broiler chicks as affected by dietary arginine. AB - A dietary deficiency of Arg may suppress chick immune system functions; however, research evaluating immune function responsiveness of commercial broilers fed dietary Arg levels near NRC (1994) recommendations is sparse. Therefore, three experiments were conducted to evaluate growth and immunity of broilers fed varying Arg levels near NRC (1994) specifications. Because Arg and Lys are similar in structure and are known to compete in intestinal absorption, dietary Lys treatments [near NRC (1994) recommendations] were evaluated to determine if Arg and Lys interact to affect broiler immunity. There were four dietary treatments in Experiment 1 representing a 2 x 2 factorial design of additional Arg (120% of NRC) or additional Lys (120% of NRC) added to a control diet containing 100% of NRC Arg and Lys (six replications per treatment). Experiment 2 contained the following four treatments: the control diet; the control diet plus L-Arg (0.20% Arg of diet); the control diet plus L-Lys HCl (0.20% Lys of diet); and the control diet plus L-Arg-L-Glu (0.10% Arg of diet). Graduations of Arg were fed from 90 to 120% of NRC in 10% increments in Experiment 3. Also, half of the birds were exposed to vaccinations of Newcastle disease virus and infectious bronchitis virus in Experiment 3 to derive a 2 x 4 factorial design. Experiments 1 and 2 were conducted from Days 1 to 18 and Experiment 3 was conducted from Days 1 to 15 in Petersime battery brooders. No interactions occurred between dietary Lys and Arg in Experiment 1. Increasing dietary Arg, but not Lys, from 100 to 120% of the NRC recommendation increased (P < or = 0.05) Day 18 BW gain. Treatment differences in the cutaneous basophil hypersensitivity assay in Experiment 1 did not occur. In Experiment 2, treatment differences in growth responses, lymphoid organ development, and primary antibody titers to SRBC did not occur. Unvaccinated birds in Experiment 3 fed an Arg-deficient diet had lower (P < or = 0.05) feed conversion in comparison with vaccinated birds fed an Arg deficient diet. Vaccinated birds had lower (P < or = 0.05) Day 15 BW than unvaccinated birds, but higher (P < or = 0.05) titers to Newcastle disease virus. Increasing dietary Arg in Experiment 3 increased plasma Arg (P < or = 0.05), but did not affect plasma Lys. Although increased dietary Arg improved BW gain in Experiment 1, minimal effects were noted in growth and immune system parameters throughtout this study. A dietary Arg level near the NRC (1994) recommendation should support proper immune system functions in healthy chicks. PMID- 11732669 TI - Characterization of alkaline hydroxide-preserved whole poultry as a dry byproduct meal. AB - Studies were conducted to examine the chemical preservation of whole broiler carcasses by using aqueous alkaline hydroxide solutions. Conversion of the preserved carcasses and solutions into an acceptable poultry byproduct meal was examined. Carcasses and alkaline solutions at a 1:1 ratio were blended and freeze dried to produce a high fat whole poultry byproduct meal. The dry meal was analyzed for nutrient composition, true metabolizable energy, and amino acid content. Viable bacteria were not recovered after inoculation of the experimental meal with Salmonella enteritidis. The meal was incorporated at 5 and 10% of chick starter diets. Chicks found the meal-containing diets acceptable. Feed consumption, water consumption, BW, and mortality were not significantly different among the dietary treatments in either of the two feeding trials. Necropsy samples revealed no pathological or histological differences attributable to consumption of the alkaline poultry byproduct and blood serum evaluation found no variation in blood chemistry. Alkaline treatment of whole broiler carcasses was an effective preservation method and acceptable as a dry poultry byproduct meal. PMID- 11732670 TI - Eimeria tenella infection in chickens: effect on plasma and muscle 3 methylhistidine. AB - To assess muscle breakdown during avian coccidiosis, the level of the nonmetabolizable amino acid 3-methylhistidine (3MH) was determined in muscle and plasma from chickens infected with the cecal parasite Eimeria tenella. The change in 3MH level during infection was determined in birds, each inoculated with 0 to 200,000 sporulated oocysts. The effect of levels of parasitism was evaluated at 6 d postinoculation. The 3MH levels of plasma and muscle were determined by HPLC after derivatization with fluorescamine. Weight gains, packed cell volumes, and gross lesion scores were also determined. E. tenella infected birds with lesion scores of 3 or 4 had significantly elevated plasma and muscle 3MH, whereas infected birds with lesion scores of 0, 1, or 2 did not have elevated plasma and muscle 3MH; however, there was a linear inverse relationship between weight gain and both plasma and muscle 3MH. The results suggested that muscle breakdown, as assessed by plasma and muscle levels of 3MH, was elevated during the acute stage of E. tenella infection and was most likely associated with anorexia caused by infection. However, the correlation of 3MH levels with severity of infection was not as strong as that previously observed for E. acervulina infection, most likely due to the differences in pathology caused by the two species. PMID- 11732671 TI - Chicken major histocompatibility complex class I definition using antisera induced by cloned class I sequences. AB - Alloantisera directed against chicken class I MHC (BFIV) antigens were produced by using transfected cell lines expressing cloned BFIV sequences. The cloned BFIV sequences were from haplotypes *12, *13, and *21. Two laboratory-derived class I mutant sequences (BFIV13m126 and BFIV21m78) were developed to analyze cross reactive epitopes and to induce specific alloantisera. Antisera were tested in hemagglutination and flow cytometry assays. The antisera produced were highly specific and had minimal cross-reactivity. The antisera induced by the BF1V21m78 mutant confirmed the significance of amino acids 78 and 81 in cross-reactivity between haplotypes B*21 and B*5. The highly specific antisera were tested by hemagglutination on red blood cells of 31 different MHC haplotypes. The consistency of hemagglutination patterns and minimal cross-reactivity demonstrated that these BFIV antisera are extremely valuable in defining MHC haplotype in various chicken lines. Because of the extreme low level of recombination between the chicken class I and class II loci, identification of BFIV allele can be used to define MHC haplotype within a line. Complete identity between the transfected cell line and the chicken used to produce the antiserum is required to ensure the monospecificity. PMID- 11732672 TI - Development of a multipurpose feed enzyme analyzer to estimate and evaluate the profitability of using feed enzyme preparations for poultry. AB - Previous studies have demonstrated that a log-linear equation could accurately predict chick performance when a feed enzyme was added to a diet and that the slope of the equation provided a measure of the efficacy of different enzymes. The objective of the study was to develop a software package, a Multipurpose Feed Enzyme Analyzer (MPFEA), based on an equation designed to evaluate the profitability of using feed enzymes. A high correlation between the efficacy of different feed enzymes (B values, the slopes of the equations) and the maximal profits was obtained when feed enzymes were added to a barley-based diet (r2 = 0.99, P < 0.0005). In contrast, there was a low correlation between the B values and the maximal profits when a feed enzyme was added to different cereal-based diets (r2 = 0.61, P = 0.2171). It appeared that there is not always a close association between efficacy of an enzyme when added to different cereal-based diets and the corresponding profitability. The MPFEA was highly versatile, as any combination of inputs such as the amounts of a feed enzyme and a substituted cereal required to yield a profit level could be determined. In conclusion, the MPFEA can accurately evaluate profitability of using different feed enzymes; select the most profitable cereal for a given feed enzyme; determine the optimal amounts of a feed enzyme, a cereal, or both; and even estimate the alternate price for a feed enzyme and a cereal. It should provide a useful tool for nutritionists. PMID- 11732673 TI - Interactive effects of dietary sodium and chloride on the incidence of spontaneous cardiomyopathy (round heart) in turkeys. AB - Two pen trials were conducted in Utah, altitude 1,630 m, with Large White female turkey poults to evaluate effects of dietary Na and chloride (Cl) concentrations on live performance and deaths caused by spontaneous cardiomyopathy (STC, "round heart") from 0 to 35 d of age. A randomized block 2 x 2 factorial arrangement with four replicate pens per treatment was used. Pen size was 3.8 x 5.6 m. In each trial, four starter diets were formulated from a common basal diet (corn-soy meat) to obtain the following sodium and chloride concentrations: High Na (0.24%):High Cl (0.40%); High Na (0.24%):Low Cl (0.16%); Low Na (0.14%):High Cl (0.40%); and Low Na (0.14%):Low Cl (0.16%). Supplements were NaCl, NaHCO3, NH4Cl, or combinations. After laboratory analysis, true values of Na were slightly lower than expected, Cl was near expected values to slightly low, and K was 1.145%. Body weight and feed conversion ratio at 35 d of age were unaffected in both trials. In both experiments, STC mortality was significantly reduced by the Low Na:High Cl treatment compared to the other treatments. In conclusion, a diet containing 0.10 to 0.12% Na and 0.38 to 0.40% Cl, significantly improved poult livability by reducing STC mortality without adverse effects on poult weight or feed conversion ratio. PMID- 11732674 TI - Efficacy of phytase for increasing protein efficiency ratio values of feed ingredients. AB - Data previously reported in several studies have suggested that phytase improves amino acid digestibility in chicks. It was the objective of the current study to determine if phytase would increase the protein efficiency ratio (PER) values (g weight gain per g protein intake) for several feed ingredients fed to chicks. Six experiments were conducted and ingredients evaluated were casein, soybean meal, canola meal, cottonseed meal, peanut meal, wheat middlings, wheat bran, rice bran, defatted rice bran, meat and bone meal, and corn gluten meal. Casein was evaluated to serve as a phytate-free control ingredient. Each feedstuff was analyzed for CP and included in cornstarch:dextrose diets as the only source of protein to provide 10% dietary protein in Experiments 1 to 5 or 18% dietary protein (soybean meal) in Experiment 6. Dietary Ca and nonphytate P levels were varied among experiments and ranged from 0.95 to 1.5% Ca and 0.35 to 0.675% nonphytate P. The test ingredient diets were fed with 0 or 1,200 units of phytase/kg to New Hampshire x Columbian chicks from 8 to 17 or 20 d of age. The PER values varied greatly among ingredients, ranging from 1.4 for corn gluten meal to 4.2 for canola meal. Phytase addition had no significant effect (P > 0.10) on PER values for any of the ingredients evaluated, except for an increase for casein in one experiment. The results indicated that 1,200 U of phytase/kg did not significantly increase protein utilization of several feed ingredients as assessed by a PER chick growth assay. PMID- 11732675 TI - 2. the effect of feeding excess methionine on live performance, carcass traits, and ascitic mortality. AB - Appetite suppressant activity of dietary excess Met was investigated in a 7-wk trial for the potential to restrict feed intake and the early growth of broilers as a means of reducing the incidence of ascitic mortality. A basal starter diet meeting the minimum NRC (1994) requirements for broilers was compared with a diet providing Met above the requirement (1.86% of the diet). Excess dietary Met was fed to the broilers from 4 to 11 d. Grower and finisher diets were provided from 22 to 35 d and from 36 to 49 d. Birds in two rooms were exposed to cool temperatures to induce ascites. The temperature program for the cool rooms was as follows: 12 h at 24 C daily (0800 to 2000) and 12 h at 15 C nightly (2000 to 0800) from 14 to 49 d, whereas the other two rooms were maintained at 24 C (control rooms). Body weight gain was depressed by 20.4% when excess Met was fed from 4 to 11 d. Feed conversion was also negatively affected by the excess dietary Met. A brief recovery was achieved from 11 to 14 d upon refeeding the well-balanced basal starter diet. Full compensatory growth was achieved by 49 d when body weight was equal to that of birds fed the basal diet. Excess dietary Met did not affect the carcass traits with the exception of abdominal fat pad, which was reduced by the excess dietary Met in the control rooms, whereas the opposite was true in the cool rooms. Ascitic mortality was not altered by dietary Met, however, the total mortality was significantly reduced by excess Met in the control rooms. PMID- 11732676 TI - Relationship between the level of dietary vitamin E and the immune response of broiler chickens. AB - The relationship between the dietary level of vitamin E (VE) and the immune response of broilers was studied in three experiments. Immunity was assessed as antibody production to infectious bronchitis virus (IBV), SRBC, and Brucella abortus (BA) antigens, mitogenic response to phytohemagglutinin A (PHA) and concanavalin A (Con A), cutaneous basophil hypersensitivity (CBH) to PHA, and lipopolysaccharide induction of acute-phase proteins (APP) and heterophilia. A range of VE (0, 10, 17.5, 25, 37.5, 50, 100, and 200 IU/kg) levels were supplemented to a basal diet (corn-soy) containing 10.2 IU of VE/kg. We found a dose-dependent increase in antibody production in response to attenuated IBV between 0 and 25 IU/kg of supplemented VE and no further increase at higher levels. Antibody levels to SRBC were higher in birds supplemented with 50 IU of VE/kg compared to those supplemented with 0 or 200 IU/kg of VE. Antibody production in response to BA antigens was not influenced by VE. Mitogenic responses were suppressed by supplemented VE in Experiment 1 for PHA (25 IU/kg diet) and Con A (25 and 50 IU/kg diets). CBH and APP levels were not affected by VE. Heterophilia was lowest at 50 IU/kg 6 h after lipopolysaccharide injection (Experiment 1). Our study showed that moderate (25 to 50 IU/kg) levels of VE supplementation were most immunomodulatory and that high levels were less effective. PMID- 11732677 TI - Additivity and associative effects of metabolizable energy and amino acid digestibility in barley and canola meal for White Pekin ducks. AB - An experiment was conducted using the TMEn bioassay method to investigate the additivity and associative effects of metabolizable energy and amino acid digestibility in barley and canola meal for White Pekin ducks. Additivity was tested by comparing the difference between observed values determined in a complete diet and predicted values from measurements determined with individual ingredients (barley and canola meal). Six ducks each were assigned to diets of barley, canola meal, the complete diet, and dextrose. Dextrose-fed ducks were used for estimation of endogenous losses for calculation of true amino acid digestibility. The observed AME, TME, AMEn, and TMEn values in the complete diet were 0.065, 0.083, 0.016, and 0.023 (kcal/g), respectively, numerically higher than predicted values. Differences between observed and predicted values were not significant (P > 0.05), indicating that the AME, AMEn, TME, and TMEn in barley and canola meal were all additive. In general, observed values for apparent amino acid digestibility (AAAD) and true amino acid digestibility (TAAD) in the complete diet were higher than those predicted from individual ingredients. Observed AAAD for lysine, histidine, tryptophan, alanine, and aspartate were higher (P < 0.05) than predicted values, indicating that digestibilities of these amino acids were not additive. The mean of AAAD in canola meal (77.29%) was higher (P < 0.05) than the observed values of barley (52.2%) and complete diet (64.55%). For TAAD values, differences between observed and predicted values were significant for lysine, histidine, and tryptophan (P < 0.05). The mean of TAAD in canola meal, barley, and complete diet were 85.88, 80.87, and 81.33%, respectively. The average difference between observed and predicted values for TAAD (1.18 %) was smaller than that of AAAD (5.41%). These results indicated that ME values for barley and canola meal were additive in the complete diet but that digestibilities of some amino acids were not additive; they further suggested that there were some associative effects of amino acids in barley and canola meal for ducks. PMID- 11732678 TI - Taurine, cardiopulmonary hemodynamics, and pulmonary hypertension syndrome in broilers. AB - Previous studies have suggested cardiac taurine is released into the plasma in response to hypoxemia (low blood oxygen levels) during the pathogenesis of pulmonary hypertension syndrome (PHS, ascites). In the present study, broilers reared under cool temperature conditions (16 C) were provided tap water (control group), tap water supplemented with taurine, or tap water supplemented with the taurine transport antagonist beta-alanine. When compared with control values, taurine supplementation consistently elevated free taurine concentrations in the plasma but not in cardiac tissues, whereas beta-alanine supplementation consistently reduced free taurine concentrations in cardiac tissues but not in the plasma. Neither the incidence of PHS nor specific predictors of PHS susceptibility (electrocardiogram Lead II S-wave amplitude, % saturation of hemoglobin with oxygen, heart rate, right to total ventricular weight ratio) were affected by taurine or beta-alanine supplementation. Cardiopulmonary hemodynamic evaluations were conducted to compare control and beta-alanine supplemented broilers breathing room air or air containing 12% oxygen (low oxygen challenge). While breathing room air, the betaalanine-supplemented broilers had higher baseline values for cardiac output (186.2 vs. 146.9 mL/min/kg BW) and pulmonary arterial pressure (27.4 vs. 22.4 mm Hg), similar values for mean systemic arterial pressure (100 vs. 104 mm Hg) and pulmonary vascular resistance (0.062 vs. 0.064 resistance units), and lower values for total peripheral resistance (0.228 vs. 0.296 resistance units) when compared with control broilers breathing room air. During low oxygen challenges, the beta-alanine-supplemented broilers exhibited larger reductions in cardiac output, mean systemic arterial pressure, and pulmonary arterial pressure and greater increases in pulmonary vascular resistance than control broilers. These observations indicate that beta-alanine supplemented broilers breathing room air had a higher systemic demand for oxygen as evidenced by their lower total peripheral resistance (systemic vasodilation) and had a capacity sufficient to pump a higher cardiac output and, thereby, maintain a similar mean systemic arterial pressure when compared with control broilers. However, cardiac function rapidly deteriorated in beta-alanine supplemented broilers during low oxygen challenges, leading to substantially greater reductions in cardiac output, stroke volume, and mean systemic arterial pressure when compared with control broilers. Concurrent changes in pulmonary arterial pressure within the beta-alanine group reflect interactions between cardiac output and pulmonary vascular resistance. Overall, depleting cardiac taurine did not appear to initiate PHS, but systemic hypoxemia developing during the mid- to late-pathogenesis of PHS may expose and incipient cardiac weakness attributable to depleted taurine reserves. PMID- 11732679 TI - Uptake of DL-2-hydroxy-4-methylthio-butanoic acid (DL-HMB) in the broiler liver in vivo. AB - The methionine source DL-2-hydroxy-4methylthio-butanoic acid (DL-HMB; Alimet feed supplement) is widely used in the poultry industry. The purpose of this study was to determine the capacity of the broiler liver to remove DL-HMB from the circulation. Cannulae were implanted in the carotid artery and hepatic and hepatic portal veins in anesthetized male broilers (3.33 +/- 0.13 kg BW). In Experiment 1, birds (n = 5) were infused with DL-HMB solutions (diluted in saline, pH 7.2 to 7.4) into the hepatic portal vein at rates ranging from 4.4 to 22 mg/min per kg BW, whereas in Experiment 2, birds (n = 6) were infused with DL HMB at rates ranging from 2.2 to 4.4 mg/min per kg BW. Plasma samples from each vessel were obtained before and after each 10-min DL-HMB infusion period with a 10-min clearance period allowed between each DL-HMB infusion. Regression analysis revealed a highly significant correlation in the amount of DL-HMB entering the liver via afferent vessels (afferent DL-HMB) and DL-HMB removed by the liver (y = 0.86(x) - 173, r2 = 0.98). The slope of this regression indicates that 86% of DL HMB entering in afferent blood (i.e. from both the hepatic artery and hepatic portal vein) was removed or that the liver apparently metabolized 86% of the DL HMB that entered the liver. The results indicate that the broiler liver has the capacity to remove DL-HMB from the circulation far in excess of that needed to metabolize DL-HMB that would enter the liver following gastrointestinal absorption in birds fed a conventional poultry diet. In addition, present results implicate the liver as a major site of removal from circulation and further metabolism of DL-HMB in chickens. PMID- 11732680 TI - The use of near-infrared reflectance spectroscopy in the prediction of the chemical composition of goose fatty liver. AB - The use of near-infrared reflectance spectroscopy (NIRS) on a meat product is described in this report. The aim of the study was to develop calibration equations to predict the chemical composition of goose fatty liver (foie gras) with lipid contents greater than 40% of the fresh pate. Spectra of 52 foie gras samples were collected in the visible and NIR region (400 to 2,498 nm). Calibration equations were computed for DM, CP, lipids and fatty acids using modified partial least-squares regression. R2 values were high for the total lipid content (0.805) and DM (0.908) but were low for ash (0.151) and relatively low for protein content (0.255). For the major fatty acids, R2 ranged from 0.886 for palmitic acid to 0.988 for oleic acid. Oleic acid, the main fatty acid of the liver, and the stearic acid had higher R2 values than the less represented fatty acids. This study suggests that the NIRS technique can be used to predict lipid content and the fatty acid composition of goose fatty livers, but calibration must be built on a larger number of samples to generate accurate predictions. PMID- 11732682 TI - Unheated water in the first tank of a three-tank broiler scalder. AB - Scalding with unheated water in the first tank of a simulated three-tank scalder was tested to determine whether carcass bacteria, efficiency of feather removal, and cooked breast meat tenderness are affected as compared with carcasses scalded at the same temperature (57 C) in all tanks. This experiment was performed on 3 d using 6-wk-old broilers. On each day, eight birds per treatment were processed. During the first 40-s scalding period, one carcass was placed in approximately 24 C water. The other carcass was placed simultaneously in a scalder unit containing approximately 2,050 L of water at 57 C. Carcasses were then held out of the water for 15 s, after which both were placed for 40 s in opposite ends of the scalder containing water at 57 C. After the second scalding period, both carcasses were again removed from the water for 15 s, followed by another 40 s in the 57 C water. Total scald time was 2 min for each treatment. After picking, carcasses were rinsed with 200 mL of sterile 0.1% peptone water for 1 min. Aerobic bacteria and Escherichia coli were enumerated and incidence of salmonella was determined by standard methods. After rinsing, carcasses were eviscerated by hand and chilled for 30 min in ice slush. All carcasses were scored for the presence of feathers, and the appearance and condition of the skin were noted. Four hours postmortem, breast fillets were removed from carcasses and chilled overnight at 2 C. The next morning, breast fillets were cooked to an internal endpoint temperature of 75 to 80 C. Warner-Bratzler shear values were measured to determine tenderness. No differences were found in numbers of aerobic bacteria and E. coli, incidence of salmonellae, tenderness of cooked breast meat, or number of feathers left on carcasses. PMID- 11732681 TI - Oxidative stability of dark chicken meat through frozen storage: influence of dietary fat and alpha-tocopherol and ascorbic acid supplementation. AB - We used factorial design to ascertain the influence of dietary fat source (linseed, sunflower and oxidized sunflower oils, and beef tallow) and the dietary supplementation with alpha-tocopheryl acetate (alpha-TA) (225 mg/kg of feed) and ascorbic acid (AA) (110 mg/kg) on dark chicken meat oxidation (lipid hydroperoxide and TBA values and cholesterol oxidation product content). alpha-TA greatly protected ground and vacuum-packaged raw or cooked meat from fatty acid and cholesterol oxidation after 0, 3.5, or 7 mo of storage at -20 C. In contrast, AA provided no protection, and no synergism between alpha-TA and AA was observed. Polyunsaturated fatty acid-enriched diets (those containing linseed, sunflower, or oxidized sunflower oils) increased meat susceptibility to oxidation. Cooking always involved more oxidation, especially in samples from linseed oil diets. The values of all the oxidative parameters showed a highly significant negative correlation with the alpha-tocopherol content of meat. PMID- 11732683 TI - The effects of electrical and controlled atmosphere stunning methods on meat and liver quality of geese. AB - The objective of this research was to investigate the effects of electrical and gas stunning on the meat and liver quality characteristics of liver geese. Sixty birds were slaughtered at 12 wk of age, in groups of 20 at three different times. Thirty birds each were subjected to one of the two stunning methods. Deboned breast fillets and thigh muscles were evaluated for hemorrhaging, amount of blood in the veins, and color by using a visual grading system. Livers were removed from carcasses during evisceration and were subsequently graded. Besides external color and hemorrhaging, the degree of liver weight loss due to removal of blood vessels was also determined. The use of controlled atmosphere stunning significantly reduced the incidence of muscle hemorrhages when compared to electrical stunning. However, no significant difference was found for color or amount of blood remaining in the veins of muscles between geese receiving electrical or controlled atmosphere stunning. The hemorrhaging and color scores of livers from gas-stunned birds did not differ from those of electrically stunned birds. As for the liver weight loss caused by removal of veins and capillaries, stunning treatment had no significant effect on this parameter. These results suggest that controlled atmosphere stunning produced slightly better quality goose meat but did not improve liver quality when compared to the electrical stunning method used. PMID- 11732684 TI - Serine proteinase inhibitors from eggs and larvae of tick Boophilus microplus: purification and biochemical characterization. AB - The present study describes the purification, characterization, and comparison of serine proteinase inhibitors during the development of egg and larva phases of the tick Boophilus microplus. Samples were collected of eggs between the first day of hatching and the beginning of eclosion (defined as El, E2, and E3) and of larvae between the first day of eclosion and the infectant phase (defined as L1, L2, and L3). Crude extracts of the samples (2.5% w/v in Tris-HCI buffer) were analyzed by SDS-PAGE, and showed three major protein bands of 42, 62, and 85 kDa, differing in intensity, from E1 to L3 samples. The total protein of the larva extracts was 34% less than that of the egg extracts, while no differences in active protein were detected. The apparent dissociation constant Ki determined for trypsin was 10-fold lower from E1 to L3 samples. Serine proteinase inhibitors from tick eggs and larvae (BmTls) were purified on trypsin-Sepharose column and analyzed by SDS-PAGE. The results showed a slight difference in protein pattern, with a protein band of 20 kDa in the E1 and E2 samples which did not appear in the other samples. The Ki for neutrophil elastase was 10-fold lower in L3 than E1. BmTI reverse-phase chromatography showed two and one major peaks in egg and larva samples, respectively. The N-terminal amino acid sequence of the L3 main peak from a C8 column showed a mix of BmTIs with the major sequence AVDFDKGCVPTADPGPCKG. Changes indicated by molecular weight and inhibition activity suggest different roles for BmTIs during the development process. PMID- 11732685 TI - Mutation of lysine residues of the 78-kDa gastrin-binding protein reduces gastrin binding. AB - The 78-kDa gastrin-binding protein (GBP) is a likely target for the antiproliferative effects of gastrin/cholecystokinin receptor antagonists on colorectal carcinoma cell lines. Both the N- and C-terminal halves of the GBP bind gastrin, but the affinity of the N-terminal half for gastrin is 7.2-fold higher than the affinity of the C-terminal half. In order to define the gastrin binding sites of the GBP in greater detail, we have constructed a truncation mutant lacking residues 221-318 of the N-terminal domain and a series of point mutants in which the lysine residues in the first 220 residues of the N-terminal domain were mutated to arginine residues. The effect of these mutations on both the extent of covalent cross-linking of iodinated gastrin2,17 and on the affinity for gastrin17 was investigated. Deletion of residues 221-318 of the GBP decreased the affinity 5.5-fold and reduced, but did not abolish, the extent of covalent cross-linking. Mutation of the 17 lysines in residues 1-220 of the GBP decreased the affinity for gastrin between 1.7- and 3.5-fold and in some cases reduced, but did not abolish, the extent of covalent cross-linking. We conclude that one or more lysine residues are involved in binding of gastrin to the GBP, but that no single lysine residue is the preferred target for covalent cross-linking of iodinated gastrin2,17 to the GBP. PMID- 11732686 TI - Isolation of unguilin, a cyclophilin-like protein with anti-mitogenic, antiviral, and antifungal activities, from black-eyed pea. AB - A protein designated unguilin was isolated from seeds of the black-eyed pea (Vigna unguiculata). It possesses a molecular weight of 18 kDa and an N-terminal sequence resembling that of cyclophilins and the cyclophilin-like antifungal protein from mung beans, and was adsorbed on Affi-gel blue gel and CM-Sepharose. Unguilin exerted an antifungal effect toward fungi including Coprinus comatus, Mycosphaerella arachidicola, and Botrytis cinerea. In addition, unguilin was capable of inhibiting human immunodeficiency virus-1 reverse transcriptase and the glycohydrolases a- and beta-glucosidases which are involved in HIV infection. Unguilin was devoid of lectin and ribonuclease activities. It inhibited methyl-3H thymidine uptake by mouse splenocytes and it weakly inhibited translation in a rabbit reticulocyte lysate system. Unguilin resembles mungin in some aspects, but differs from it in others. PMID- 11732687 TI - A monomeric mannose-binding lectin from inner shoots of the edible chive (Allium tuberosum). AB - A mannose-binding lectin was isolated from the inner shoots of the chive Allium tuberosum. The procedure involved aqueous extraction, (NH4)2SO4 precipitation, dialysis to remove (NH4)2SO4, affinity chromatography on mannose-agarose, ion exchange chromatography on SP-Sepharose, gel filtration on Superdex 75, and ion exchange chromatography on Mono S. Lectin activity was adsorbed on mannose agarose, SP-Sepharose, and Mono S. The lectin demonstrated a molecular weight of 13 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, indicating that it is a single-chain protein. N-terminal sequence analysis revealed its remarkable homology to Allium cepa lectin and similarity to a lesser extent to lectins from members of the Amaryllidaceae, Orchidaceae, and Liliaceae. The lectin manifested mitogenic activity in murine splenocytes and inhibitory activity against human immunodeficiency virus type 1 reverse transcriptase. PMID- 11732688 TI - Isolation of a homodimeric lectin with antifungal and antiviral activities from red kidney bean (Phaseolus vulgaris) seeds. AB - A homodimeric lectin adsorbed on Affi-gel blue gel and CM-Sepharose and possessing a molecular weight of 67 kDa was isolated from red kidney beans. The hemagglutinating activity of this lectin was inhibited by glycoproteins but not by simple sugars. The lectin manifested inhibitory activity on human immunodeficiency virus-1 reverse transcriptase and alpha-glucosidase. The N terminal sequence of the lectin exhibited some differences from previously reported lectins from Phaseolus vulgaris but showed some similarity to chitinases. It exerted a suppressive effect on growth of the fungal species Fusarium oxysporum, Coprinus comatus, and Rhizoctonia solani. The lectin had low ribonuclease and negligible translation-inhibitory activities. PMID- 11732689 TI - Assignment of the disulfide bridges in bothropstoxin-I, a myonecrotic Lys49 PLA2 homolog from Bothrops jararacussu snake venom. AB - Bothropstoxin-I (BthTX-I), a Lys49 phospholipase A2 homolog with no apparent catalytic activity, was first isolated from Bothropsjararacussu snake venom and completely sequenced in this laboratory. It is a 121-amino-acid single polypeptide chain, highly myonecrotic, despite its inability to catalyze hydrolysis of egg yolk phospholipids, and has 14 half-cystine residues identified at positions 27, 29. 44. 45, 50, 51, 61, 84, 91, 96, 98, 105, 123, and 131 (numbering according to the conventional alignment including gaps, so that the last residue is Cys 131). In order to access its seven disulfide bridges, two strategies were followed: (1) Sequencing of isolated peptides from (tryptic + SV8) and chymotryptic digests by Edman-dansyl degradation; (2) crystallization of the protein and determination of the crystal structure so that at least two additional disulfide bridges could be identified in the final electron density map. Identification of the disulfide-containing peptides from the enzymatic digests was achieved following the disappearance of the original peptides from the HPLC profile after reduction and carboxymethylation of the digest. Following this procedure, four bridges were initially identified from the tryptic and SV8 digests: Cys5O-Cysl31, Cys51-Cys98, Cys61-Cys91, and Cys84-Cys96. From the chymotryptic digest other peptides were isolated either containing some of the above bridges, therefore confirming the results from the tryptic digest, or presenting a new bond between Cys27 and Cys123. The two remaining bridges were identified as Cys29-Cys45 and Cys44-Cys105 by determination of the crystal structure, showing that BthTX-I disulfide bonds follow the normal pattern of group II PLA2s. PMID- 11732690 TI - Purification and characterization of a variant of rhodocetin from Calloselasma rhodostoma (Malayan pit viper) venom. AB - Rhodocetin is a novel C-type lectin-related protein (CLP) purified from the venom of Calloselasma rhodostoma. Thus far, it is the only reported CLP whose alpha and beta subunits are not linked by an interdisulfide bond. We report here the isolation of a variant of rhodocetin from a different source of venom. This variant of rhodocetin exhibited a different elution profile in reverse-phase HPLC as compared to the rhodocetin reported in our original publication [Wang et al., (1999), Biochemistry 38, 7584-7593]. Specifically, the alpha subunit of the variant was eluted at a considerably lower percentage of acetonitrile, which suggested a less hydrophobic polypeptide chain as compared to the original rhodocetin. Using a combination of microcharacterization techniques such as peptide mapping, mass spectrometry, and amino acid sequencing, we identified an amino acid substitution, 163K, in the polypeptide chain that could account for the difference in elution behavior of the alpha subunit. In addition, we also found a conserved E88D substitution in the beta chain which was not apparent during reverse-phase HPLC. However, neither of these substitutions resulted in the alteration of the functional properties of the rhodocetin variant. PMID- 11732691 TI - Solution structures of casein peptides: NMR, FTIR, CD, and molecular modeling studies of alphas1-casein, 1-23. AB - To determine its potential for interacting with other components of the casein micelle, the N-terminal section of bovine alphas1-casein-B, residues 1-23, was investigated with nuclear magnetic resonance (NMR), Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopies, and molecular modeling. NMR data were not consistent with conventional alpha-helical or beta-sheet structures, but changes in N-H proton chemical shifts suggested thermostable structures. Both CD and FTIR predicted a range of secondary structures for the peptide (30-40% turns, 25-30% extended) that were highly stable from 5 degrees C to 25 degrees C. Other conformational elements, such as loops and polyproline II helix, were indicated by FTIR only. Molecular dynamics simulation of the peptide predicted 32% turns and 27% extended, in agreement with FTIR and CD predictions and consistent with NMR data. This information is interpreted in accord with recent spectroscopic evidence regarding the nature of unordered conformations, leading to a possible role of alphas1-casein (1-23) in facilitating casein-casein interactions. PMID- 11732692 TI - Amine oxidase from lentil seedlings: energetic domains and effect of temperature on activity. AB - Copper/TPQ amine oxidases from mammalian and plant sources have shown many differences in substrate specificity and molecular properties. In this work the activity of lentil seedling amine oxidase was followed at various temperatures in 100 mM potassium phosphate buffer, pH 7, using benzylamine as substrate. The discontinuous Arrhenius plot of lentil amine oxidase showed two distinct phases with a jump between them. Thermal denaturation of the enzyme, using differential scanning calorimetry under the same experimental conditions, showed a transition at the same temperature ranges in the absence of substrate, indicating the occurrence of conformational changes, with an enthalpy change of about 175.9 kJ/mole. The temperature-induced changes of the activity of lentil amine oxidase are compared with those of bovine serum amine oxidase (taken from the literature). PMID- 11732693 TI - Expression of A chain and B chain of beta-bungarotoxin from taiwan banded krait: the functional implication of the interchain disulfide bond between A chain and B chain. AB - beta-Bungarotoxin (beta-Bgt), the main presynaptic neurotoxin purified from the venom of Bungarus multicinctus, consists of two dissimilar polypeptide chains, the A chain and B chain, cross-linked by an interchain disulfide bond. The A and B chain cDNAs were subcloned into expression vectors pT7-7 and pET20b(+), respectively, and transformed into Escherichia coli strain BL21(DE3). The expressed protein was isolated from the inclusion bodies of E. coli and subjected to refolding into its folded structure. The yields of the refolded A and B chains increased markedly by at least 100-fold after substituting Ser for Cys1S of A chain and Cys55 of B chain, which formed an interchain disulfide bond. Either the A(C15) chain or B(C55S) chain alone or in combination cannot exhibit the phospholipase A2 activity or synaptosome binding activity of beta-Bgt. Nevertheless, the results of competitive enzyme-linked immunoassay, CD spectra, and fluorescence measurement revealed that the A(C15S) chain and B(C55S) chain possessed a native-like structure like the subunits of native beta-Bgt. Moreover, the interfacial interaction between the A and B chains explored by glutaraldehyde cross-linking revealed the essential aspects of the intact interchain disulfide bond in this interaction. This suggests that the formation of the interchain disulfide bond should not be a crucial step for the formation of folded A and B chains in the venom glands, and that the integrity of the interchain disulfide linkage favors the subunit interaction that consequently fulfills the functional mechanism of beta-Bgt. PMID- 11732694 TI - Crystal structure of the complex of concanavalin A and hexapeptide. AB - The X-ray structure analysis of a cross-linked crystal of concanavalin A soaked with a hexapeptide molecule as a probe molecule showed an electron density corresponding to full occupation in the binding pocket. The site lies on the surface of concanavalin A and is surrounded by three symmetry-related molecules. The crystal structure of the hexapeptide complex was refined at 1.93-A resolution, to an R-factor of 19% (Rfree factor of 25%), with an RMS deviation in bond distances of 0.01 A. The model includes all 237 residues of concanavalin A, one manganese ion, one calcium ion, 95 water molecules, one glutaraldehyde molecule, one isopropanol molecule, and one hexapeptide molecule. This X-ray structure analysis also provides an approach to mapping the binding surface of crystalline protein with a probe molecule that is dissolved in the mixture of organic solvent with water or in neat organic solvent but is hardly dissolved in aqueous solution. PMID- 11732695 TI - Urinary thioether excretion and erythrocyte glutathione S-transferase activities in smokers and patients with squamous-cell laryngeal cancer. AB - Various biomarkers exist for assessment of exposure to cigarette smoke. The aim of this study is to evaluate whether the urinary thioethers (UT) and erythrocyte glutathione S-transferase (GST) enzyme activities can be used as a biomarker or indicator of a higher risk of laryngeal cancer among smokers. In the present study, the concentration of UT and erythrocyte CST activities were measured in a sample of 84 subjects: controls, smokers, and smokers with squamous-cell carcinoma of the larynx (SLC). For this study, cases were restricted to men, since the number of women was not sufficient for statistical comparisons. Smoking significantly increased UT levels without a marked change in GST activity. However, in SLC smoking patients both UT levels and GST activity were significantly elevated. The use of nonspecific UT levels with GST activity determination seems to be a reliable indicator for the presence of laryngeal cancer in smokers. PMID- 11732696 TI - Characterization of age-related changes in body weight and organ weights from birth to adolescence in humans. AB - The pharmacokinetics and tissue dose of chemicals may differ among individuals of a population, particularly between adults and children. The adult-children differences in pharmacokinetics arise from age-related changes in the physiological, biochemical, and physicochemical determinants of uptake and disposition of chemicals. The objectives of this study were to review the published literature to assemble data on the human body weight and organ weights as a function of age (specifically between birth and 18 yr old) and to analyze these data, in order to develop regression equations for calculating body weight and organ weights of children using age as the dependent function. The specific organs/tissues for which the data on age-related weight were obtained and analyzed include blood, adipose tissues, liver, lungs, brains, heart, kidneys, spleen, the reproductive organs (male: prostate gland, seminal vesicle, testes, and epididymis; female: ovaries, uterus, and uterine tubes), glands (adrenal, pituitary, thymus, pancreas, and thyroid), bone marrow (total and red), intestinal tract, stomach, muscle, skin (epidermis and dermis), and skeleton. In both male and female children, the sum of these organs is systematically lower than the body weight, and this discrepancy may be resolved with the additional availability and consideration of data on hypodermis weight. The equations and data on body weight and organ weights presented in this article should be useful for constructing age-specific, physiologically based pharmacokinetic models for children. PMID- 11732697 TI - Betel quid chewing and risk of adverse birth outcomes among aborigines in eastern Taiwan. AB - It is known that substance abuse during pregnancy is associated with increased risk of adverse birth outcomes. The aim of this study was to determine the use of alcohol, cigarettes, betel quid, and drugs among pregnant aboriginal women and to assess the risk of adverse effects of betel quid use on birth outcomes in eastern Taiwan. Of a total of 229 women recruited into this study, 32 women with adverse birth outcomes constituted the case group. Analyses revealed that adverse birth outcomes were associated with maternal betel quid chewing and maternal age. After adjusting for maternal age, the risk of adverse birth outcome was five times higher among betel quid chewing women as compared to substance nonusers. Based on this finding, it is suggested health education, especially when concerned with the harmful effects of substance abuse, which includes betel quid use during pregnancy, should be stressed in concert with routine prenatal care. PMID- 11732698 TI - Analyses of micronuclei in exfoliated epithelial cells from individuals chronically exposed to arsenic via drinking water in inner Mongolia, China. AB - The groundwater in Bayingnormen (Ba Men), located in Central West Inner Mongolia, China, is naturally contaminated with arsenic at concentrations ranging from 50 microg/L to 1.8 mg/L. Various adverse health effects in this region, including cancer, have been linked to arsenic exposure via drinking water. A pilot study was undertaken to evaluate frequencies of micronuclei (MN), as measures of chromosomal alterations, in multiple exfoliated epithelial cell types from residents of Ba Men chronically exposed to arsenic via drinking water. Buccal mucosal cells, airway epithelial cells in sputum, and bladder urothelial cells were collected from 19 residents exposed to high levels of arsenic in drinking water (527.5 +/- 24 microg/l), and from 13 control residents exposed to relatively low levels of arsenic in drinking water (4.4 +/- microg/L). Analytical results from these individuals revealed that MN frequencies in the high-exposure group were significantly elevated to 3.4-fold over control levels for buccal and sputum cells, and to 2.7-fold over control for bladder cells (increases in MN frequency significant at p < .001 for buccal cells; p < .01 for sputum cells; p < .05 for bladder cells). When smokers were excluded from high-exposure and control groups the effects of arsenic were observed to be greater, although only in buccal and sputum cells; approximately 6-fold increases in MN frequency occurred in these tissues. The results indicate that residents of Ba Men chronically exposed to high levels of arsenic in drinking water reveal evidence of genotoxicity in multiple epithelial cell types; higher levels of induced MN were observed in buccal and sputum cells than in bladder cells. PMID- 11732699 TI - Polychlorinated biphenyls in blood plasma among Swedish female fish consumers in relation to time to pregnancy. AB - The purpose of this study was to assess the association between 2,2',4,4',5,5' hexachlorobiphenyl (CB-153) in plasma, a biomarker of exposure to polychlorinated biphenyls (PCB), and time to pregnancy (TTP) in a group of women with a varying dietary exposure to PCB. For 121 Swedish east coast fishermen's wives (median year of birth 1956, range 1945-1968), information on selt-reported TTP for the first planned pregnancy (median 2 mo, range 0-48) and CB-153 concentrations from blood samples drawn in 1995 (median 144 ng/g lipid, range 16-566) were available. Each woman's CB-153 concentration in plasma at the time immediately preceding her pregnancy was estimated, taking into account reduction of body burden of CB-153 due to lactation, the yearly reduction of PCB in Baltic Sea fish, as well as the biological half-life of CB-153. Based on the estimated CB-153 concentrations, subjects were categorized into tertiles as low (37-206 ng/g lipid), medium (207 330 ng/g lipid), and high (331-,1036 ng/g lipid) exposure groups. TTP in the medium- and high-exposure groups were then compared to TTP in the low-exposure group by estimating the corresponding success rate (i.e., the number of pregnancies per person month) ratios (SuRR) using discrete Cox regression, taking into account essential confounders. No obvious association between estimated CB 153 concentration and TTP was observed (medium vs. low: SuRR 0.77 [95% CI 0.47 1.28] and high vs. low: SuRR 0.95 10.74-1.23]). The present data give no support for a negative association between the plasma CB-153 concentrations observed in the present study and TTP. It should, however, be borne in mind that the study group was rather small and mainly included relatively young women, likely to have been only moderately exposed. PMID- 11732700 TI - Oxidative stress in heart tissue of hyperthyroid and iron supplemented rats. AB - This study was designed to investigate the effect of hyperthyroidism and/or iron supplementation or cardiac oxidative stress parameters--the lipid peroxidation end product glutathione (GSH), glutathione peroxidase (CSH-Px), and superoxide dismutase (CuZnSOD)--in rats. In plasma, ferritin as an indicator of iron status and glutamate oxaloacetate transaminase (GOT) as an indicator of damage to the heart tissue were analyzed. Our findings show that hyperthyroidism increased lipooxidative damage as reflected by higher lipid peroxidation end product levels and elevated antioxidant defense parameters-GSH and GSH-Px. Iron supplementation per se does not affect oxidative stress parameters studied in the euthyroid state. Although iron increased lipid peroxidation in the hyperthyroid state, this effect was less than that seen in euthyroidism. Iron supplementation to hyperthyroid rats significantly lowered plasma ferritin levels, suggesting increased iron elimination with consequently reduced oxidative stress. PMID- 11732701 TI - Critical role of reactive oxygen species formation in microcystin-induced cytoskeleton disruption in primary cultured hepatocytes. AB - Cyanobacteria (blue-green algae)-contaminated water is a worldwide public health problem. Microcystins are a group of liver-specific toxins generated by cyanobacteria. It is generally believed that the protein phosphorylation that leads to the disruption of intermediate filaments plays an important role in microcystin-induced hepatotoxicity. However, the mechanisms that contribute to the microcystin-induced alterations of microtubules and microfilaments are not fully understood. In the present study, the effects of microcystin-fR (M-LR), the most common microcystin, were examined on the organization of cellular microtubules and microfilaments in primary cultured rat hepatocytes. Our results indicate that M-LR initiated reactive oxygen species (ROS) formation followed by altering the cytoskeleton structures, which eventually led to significant LDH leakage. These effects were completely prevented by TEMPOL, a superoxide dismutase mimic, and also partially prevented by desferoxamine. These findings provide further evidence that ROS formation, especially superoxide radical, plays a crucial role in M-LR-induced disruption of cytoskeleton organization and consequent hepatotoxicity. PMID- 11732702 TI - GluR5,6,7 subunit immunoreactivity on apical pyramidal cell dendrites in hippocampus of schizophrenics and manic depressives. AB - Recent postmortem studies have suggested that changes in the regulation of kainate-sensitive glutamate receptors (kainate receptors) in the hippocampus may play a role in schizophrenia. To explore this possibility further, the distribution of immunoreactivity (IR) for the GluR5,6,7 subunits of the KR was assessed in a cohort consisting of 15 normal controls, 15 schizophrenics, and 9 manic depressives matched for age and postmortem interval (PMI). Cross sections of hippocampus showed abundant GluR5,6,7-IR on apical dendrites of pyramidal neurons in the stratum radiatum and stratum moleculare. In normal controls, both the numerical and length density of IR dendrites were much higher in sector CA2 than in sectors CA3 or CA1. When data for the individual groups were separately examined, the schizophrenics showed a 30-35% reduction in the density of GluR5,6,7-IR dendrites found in both stratum radiatum and stratum moleculare of sectors CA3 and CA2, as well as proximal and middle portions of CA1. In CA2, the magnitude of this decrease in schizophrenia was 2.5 times larger than that seen in any of the other sectors. For the manic depressive group, no significant differences were observed in any sectors or laminae examined. The potential confounding effects of either age, PMI, or neuroleptic exposure do not explain the reduced density of IR dendrites detected in the schizophrenic group. Taken together, the preferential reduction of GluR5,6,7-IR observed on apical dendrites of pyramidal neurons is consistent with a functional downregulation of the kainate receptor in the hippocampus of schizophrenic brain. PMID- 11732703 TI - Effects of pre- and postnatal corticosterone exposure on the rat hippocampal GABA system. AB - Several lines of evidence have implicated prenatal stress and the hippocampal GABA system in the pathophysiology of schizophrenia, and prenatal stress is believed to increase the risk for schizophrenia through alterations of this neurotransmitter. To explore this hypothesis, we treated male rats pre- and/or postnatally (P48 and P60) with either corticosterone (CORT) or vehicle to establish three study groups: VVV, receiving vehicle at all three time points; VCC, receiving vehicle prenatally and CORT at both postnatal timepoints; and CCC, receiving CORT at all three timepoints. Animals were sacrificed at either 24 h or 5 days after final injection and examined for mRNA levels of GAD65, GAD67, and the GABA(A) receptor subunits alpha2 and gamma2. At 24 h, GAD65 mRNA was decreased in CA1, CA2, CA4, and dentate gyrus (DG) of VCC rats; this effect was either decreased or reversed in CCC-treated animals. No effect was detected in GAD67 mRNA at 24 h. At 5 days, CORT treatment increased GAD67 mRNA levels in CA1, CA3, and DG. Prenatal treatment with CORT was associated with increased responsiveness only in CA3 and DG. For the GABAA receptor, alpha2 subunit mRNA did not show any change in response to CORT treatment, while that for the gamma2 subunit was decreased in CA2 of both VCC- and CCC-treated animals. Consistent with gamma2 subunit mRNA decreases, benzodiazepine (BZ) receptor binding activity was decreased in CA2 with CORT treatment. Prenatal CORT exposure neither increased nor decreased this effect. These results demonstrate that CORT administration is associated with a complex regulation of mRNA expression for pre and postnatal aspects of the hippocampal GABA system. Under these conditions, prenatal exposure to CORT may sensitize some of these effects, but does not fundamentally alter the nature of this response. PMID- 11732704 TI - Neuropathological studies of synaptic connectivity in the hippocampal formation in schizophrenia. AB - Cytoarchitectural changes in the hippocampal formation have been prominent among the various neuropathological abnormalities reported in schizophrenia. Replicated positive findings include decreased neuronal size and alterations in presynaptic and dendritic markers. These findings, in the absence of neurodegenerative changes, suggest that there are alterations in the neural circuitry in schizophrenia. These may represent the anatomical correlate of the aberrant functional connectivity described in neuroimaging studies, which in turn contributes to the psychotic and cognitive symptomatology of the disorder. The identity of the affected hippocampal circuits remains unclear; there is evidence for both glutamatergic and GABAergic involvement, and perhaps for a gradient of pathology in which changes are most apparent in CA4 and the subiculum, and least in CA1. The data, their interpretation, and their limitations are discussed, with particular emphasis upon molecular and immunological studies of synaptic protein gene expression. PMID- 11732705 TI - Neuroimaging studies of the hippocampus in schizophrenia. AB - Three neuroimaging techniques, morphometric neuroimaging, magnetic resonance spectroscopy, and functional neuroimaging, have provided evidence for abnormal hippocampal structure and function in schizophrenia. Hippocampal volume reduction is now one of the most consistent structural abnormalities found in schizophrenia: it is present at the onset of the illness and, to a lesser degree, in first-degree relatives of schizophrenic probands. Decreased levels of N-acetyl aspartate point towards a cellular basis of such volume changes. Functional neuroimaging studies have demonstrated abnormal levels of hippocampal activity at rest, during the experience of auditory hallucinations, and during the performance of memory retrieval tasks. These results of neuroimaging studies complement evidence from post-mortem and behavioral studies, which have found regionally specific abnormalities of the hippocampus and of memory function in schizophrenia. PMID- 11732706 TI - NMDA receptor regulation of memory and behavior in humans. AB - N-methyl-D-aspartate (NMDA) receptor hypofunction is associated with a range of effects on cognition and behavior in whole animal and human studies. NMDA receptor hypofunction within the brain, which can be induced experimentally in vivo using NMDA receptor antagonist drugs, produces adverse effects on memory function. The results suggest that NMDA receptor hypofunction can preferentially affect neural mechanisms regulating the efficiency of encoding and consolidation into longer-term storage. More pronounced NMDA receptor hypofunction can produce a clinical syndrome that includes core features of psychosis, as well as dissociation. Finally, sustained and severe underexcitation of NMDA receptors in the adult brain is associated with a neurotoxic process with well-characterized neuropathological features. Progressive increases in severity of NMDA receptor hypofunction within the brain can produce a range of effects on brain function, involving local and distributed circuitry, which may underlie the observed changes in behavior. As the brain ages, the NMDA receptor system becomes progressively hypofunctional, potentially contributing to further age-related decreases in memory and learning performance. Pharmacological and genomic methods for preventing NMDA receptor hypofunction, or for preventing the upstream or downstream consequences modeled by treatment with NMDA antagonists, may be applicable to the prevention and treatment of memory and behavioral dysfunction in a variety of neuropsychiatric disease conditions. PMID- 11732707 TI - Probing the human hippocampus using rCBF: contrasts in schizophrenia. AB - Regional cerebral blood flow (rCBF) data from two PET-15O water schizophrenia studies were analyzed using individually placed, magnetic resonance (MR)-guided hippocampal volumes of interest (VOI). In one study, normal (N = 10) and schizophrenic (N = 18) volunteers performed an overlearned auditory discrimination task in rest, control, and decision conditions. In the other study, schizophrenic and normal volunteers received the noncompetitive NMDA receptor antagonist ketamine and placebo and had sequential rCBF evaluations. Moreover, the schizophrenic volunteers were off drug in one study and on antipsychotic drug in the second study, allowing an additional comparison of medication status. VOIs were placed on anterior, middle, and posterior hippocampal areas in each PET image from both studies, redirected from an MR scan, and individually adjusted. While no hippocampal activation was apparent in either the normal or schizophrenic group in the task vs. condition comparison, rCBF was higher in the schizophrenic than in the normal hippocampus in both task and control conditions, independently. In addition, at rest rCBF was significantly higher in the unmedicated group of schizophrenics than in the group of medicated patient volunteers and higher than in the normal comparison group. This suggests that schizophrenia is associated with elevated rCBF in the hippocampus, which "normalizes" with antipsychotic drug treatment. Ketamine, the noncompetitive NMDA receptor antagonist, was more potent in reducing rCBF in the schizophrenic group compared to the normal volunteer group. These data are consistent with a previous report from our laboratory of reduced NMDA receptor NR1 subunit expression and possible abnormal NMDA receptor composition in schizophrenia. These data show an abnormality of hippocampal function in schizophrenia and suggest that this abnormality may be associated with the pathophysiology of the illness. PMID- 11732708 TI - Storage, recall, and novelty detection of sequences by the hippocampus: elaborating on the SOCRATIC model to account for normal and aberrant effects of dopamine. AB - In order to understand how the molecular or cellular defects that underlie a disease of the nervous system lead to the observable symptoms, it is necessary to develop a large-scale neural model. Such a model must specify how specific molecular processes contribute to neuronal function, how neurons contribute to network function, and how networks interact to produce behavior. This is a challenging undertaking, but some limited progress has been made in understanding the memory functions of the hippocampus with this degree of detail. There is increasing evidence that the hippocampus has a special role in the learning of sequences and the linkage of specific memories to context. In the first part of this paper, we review a model (the SOCRATIC model) that describes how the dentate and CA3 hippocampal regions could store and recall memory sequences in context. A major line of evidence for sequence recall is the "phase precession" of hippocampal place cells. In the second part of the paper, we review the evidence for theta-gamma phase coding. According to a framework that incorporates this form of coding, the phase precession is interpreted as cued recall of a discrete sequence of items from long-term memory. The third part of the paper deals with the issue of how the hippocampus could learn memory sequences. We show that if multiple items can be active within a theta cycle through the action of a short term "buffer," NMDA-dependent plasticity can lead to the learning of sequences presented at realistic item separation intervals. The evidence for such a buffer function is reviewed. An important underlying issue is whether the hippocampal circuitry is configured differently for learning and recall. We argue that there are indeed separate states for learning and recall, but that both involve theta oscillations, albeit in possibly different forms. This raises the question of how neuromodulatory input might switch the hippocampus between learning and recall states and more generally how different neuromodulatory inputs reconfigure the hippocampus for different functions. In the fifth part of this paper we review our studies of dopamine and dopamine/NMDA interactions in the control of synaptic function. Our results show that dopamine dramatically reduces the direct cortical input to CA1 (the perforant path input), while having little effect on the input from CA3. In order to interpret the functional consequences of this pathway specific modulation, it is necessary to understand the function of CA1 and the role of dopaminergic input from the ventral tegmental area (VTA). In the sixth part of this paper we consider several possibilities and address the issue of how dopamine hyperfunction or NMDA hypofunction, abnormalities that may underlie schizophrenia, might lead to the symptoms of the disease. Relevant to this issue is the demonstrated role of the hippocampus in novelty detection, a function that is likely to depend on sequence recall by the hippocampus. Novelty signals are generated when reality does not match the expectations generated by sequence recall. One possible site for computing mismatch is CA1, since it receives predictions from CA3 and sensory "reality" via the perforant path. Our data suggest that disruption of this comparison would be expected under conditions of dopamine hyperfunction or NMDA hypofunction. Also relevant is the fact that the VTA, which fires in response to novelty, may both depend on hippocampal-dependent novelty detection processes and, in turn, affect hippocampal function. Through large-scale modeling that considers both the processes performed by the hippocampus and the neuromodulatory loops in which the hippocampus is embedded, it is becoming possible to generate working hypotheses that relate synaptic function and malfunction to behavior. PMID- 11732709 TI - Circuit analysis of NMDAR hypofunction in the hippocampus, in vitro, and psychosis of schizophrenia. AB - NMDA antagonists provide the best pharmacological model of psychosis-related schizophrenia. Data from circuit analysis of the effects of the antagonism of NMDA receptors in the CA1 region of the hippocampus of rats in vitro suggest a hypothesis concerning cortical circuit dysfunction responsible for NMDA antagonist-dependent psychosis, relevant to the psychosis associated with schizophrenia. The NMDA antagonists may act by causing a selective, partial, disinhibition of cortical projection cells. The effects are partially due to the partial role of NMDA-dependent transmission in the excitatory glutamate drive of interneurons. Characterization of the selectivity is incomplete, but includes disinhibition of the recurrent inhibitory circuit and is concentration-sensitive. It may result from differences in NMDA receptors (NMDARs) on interneurons. At higher concentrations, antagonism of all NMDA-dependent transmission results in anesthesia. At low concentration, selective blockade of NMDA-dependent LTP of the recurrent inhibitory circuit may disrupt particular aspects of information processing involving learning and/or memory, consistent with the generation of abnormal associations. An endogenous peptide, NAAG, is shown to antagonize NMDARs in a manner similar to known psychotogenic agents like ketamine or phencyclidine. Finally, mechanisms that could enhance NMDAR function are discussed as possible therapeutic strategies for psychosis. PMID- 11732710 TI - Hippocampus as comparator: role of the two input and two output systems of the hippocampus in selection and registration of information. AB - Processing of multimodal sensory information by the morphological subdivisions of the hippocampus and its input and output structures was investigated in unanesthetized rabbits by extracellular recording of neuronal activity. Analysis shows principal differences between CA3 neurons with uniform multimodal, mainly inhibitory, rapidly habituating sensory responses, and CA1-subicular neurons, substantial parts of which have phasic reactions and patterned on-responses, depending on the characteristics of the stimuli. These differences result from the organization of the afferent inputs to CA1 and CA3. Analysis of neuronal responses in sources of hippocampal inputs, their electrical stimulation, and chronic disconnection show the greater functional significance of the brain-stem reticular input for tonic responses characteristic of CA3. This input signal before entering the hippocampus is additionally preprocessed at the MS-DB relay, where it becomes more uniform and frequency-modulated in the range of theta rhythm. It is shown that the new sensory stimuli produce inhibitory reset, after which synchronized theta-modulation is triggered. Other stimuli, appearing at the background of the ongoing theta, do not evoke any responses of the hippocampal neurons. Thus, theta-modulation can be regarded as a mechanism of attention, which prolongs response to a selected stimulus and simultaneously protects its processing against interference. The cortical input of the hippocampus introduces highly differentiated information analyzed at the highest levels of the neocortex through the intermediary of the entorhinal cortex and presubiculum. However, only CA1-subiculum receives this information directly; before its entrance into CA3, it is additionally preprocessed at the FD relay, where the secondary simplification of signals occurs. As a result, CA3 receives by its two inputs (MS DB and FD) messages just about the presence and level of input signals in each of them, and performs relatively simple functions of determination of match/mismatch of their weights. For this comparator system, the presence of signal only in the reticulo-septal input is equivalent to quality of novelty. The cortical signal appears with some delay, after its analysis in the neocortex and shaping in the prehippocampal structures; besides, it is gradually increased due to LTP-like incremental changes in PP and mossy fiber synapses. The CA3 neurons with potentiated synapses of cortical input do not respond to sensory stimuli; that is, the increased efficacy of the cortical signals can be regarded as "familiarity" of a signal, terminating the reactive state of the CA3 neurons. The integrity of both inputs is necessary for gradual habituation of sensory responses in the hippocampus. The output signals of CA3 following in the precommissural fornix to the output relay-LS nucleus and to the brain-stem structures have strong regulatory influence on the level of brain activity (arousal), which is an important condition for processing and registration of information. The primary targets of this output signal are raphe nuclei, which suppress activity of the ascending excitatory RF. In the background state, activity of the CA3 neurons through the intermediary of raphe keeps RF under tonic inhibitory control. Inhibition of the majority of CA3 pyramidal neurons during a novel stimulus action decreases the volume of its output signal to raphe and releases RF from tonic inhibition (increase in level of activity of the forebrain, arousal). When the responses of CA3 neurons habituate, the initial high background activity is reinstated, as well as tonic suppression of RF. Analysis of the second output of CA3 (by Schaffer's collaterals to CA1) shows that activity in this pathway can block access of cortical signals from PP to CA1 neurons by action upon the local system of inhibitory neurons, or by shunting the propagation of signals in apical dendrites. Thus, CA3 can act as a filter controlling the information transmission by CA1; such transmission at any given moment is allowed only in those CA1 neurons which receive SC from CA3 neurons, responding to the sensory stimulus by suppression of their activity. Disconnection of the CA3 output fibers results in disappearance of habituation in all its target structures (raphe, RF, CA1). The output signal of CA1-subiculum follows by postcommissural fornix to the chain of structures of the main limbic circuit: mammillary bodies (medial nucleus), anterior thalamic nuclei (mainly antero-ventral nucleus), and cingulate limbic cortex (mainly posterior area). In each of these links, the signal is additionally processed. Habituation is nearly absent in these structures; instead, st PMID- 11732711 TI - Hippocampal dysfunction in schizophrenia. AB - Although not necessarily primary to the disease, hippocampal dysfunction in schizophrenia is suggested by morphological changes in the hippocampal formation reported in schizophrenic patients. This notion receives additional support from studies showing that 1) similar behavioral deficits are exhibited by both schizophrenics and animals with hippocampal lesions, and 2) some of these behavioral deficits are reversed by neuroleptics in both schizophrenics and lesioned animals. A brain-mapped neural network model is used to explain how some impairments in attention can be caused by hippocampal dysfunction and ameliorated by dopaminergic blockers. PMID- 11732712 TI - Purification and characterization of an intracellular beta-glucosidase from a Candida sake strain isolated from fruit juices. AB - A yeast strain isolated in the laboratory from fruit juices was studied and classified as Candida sake. The strain produces an intracellular beta-glucosidase when grown with cellobiose as the carbon source. The enzyme was purified by ion exchange chromatography and gel filtration. The molecular mass of the purified intracellular beta-glucosidase, estimated by gel filtration, was 240 kDa. The tetrameric structure of the beta-glucosidase was determined following treatment of the purified enzyme with sodium dodecyl sulfate. The enzyme exhibited optimum activity at 52 degrees C and pH 4.25 with citrate-phosphate buffer. The enzyme was active against soluble glycosides with the (1-->4)-beta configuration, and from Lineweaver Burk plots, a Km value of 6.9 mmol/L was found for p-nitrophenyl beta-D-glucopyranoside. The beta-glucosidase was found to be tolerant to glucose inhibition with a Ki value of 0.2 mol/L. PMID- 11732713 TI - Fluorescence investigation of affinity interaction between bovine serum albumin and triazine dye in reversed micelles. AB - The effect of Cibacron Blue 3GA (CB) on the fluorescence emission spectra of bovine serum albumin (BSA) was investigated in cationic reversed micelles formed with cetyltrimethylammonium bromide (CTAB) compared with that in aqueous phase. The anionic CB had electrostatic interactions with cationic CTAB and affinity interactions with BSA in the reversed micelles. The addition of CB in the reversed micellar phase led to a great decrease in the fluorescence intensity of BSA and a remarkable red shift of the wavelength of emission maximum (lambda(max)). The fluorescence intensity of BSA decreased and the lambda(max) decreased 5 nm (blue shift) without the addition of CB in the reversed micellar phase. The fluorescence intensity of BSA with the addition of CB had the strongest value in the aqueous phase with the presence of CTAB, a less strong value in the reversed micellar phase, and a weak value in the aqueous phase without the presence of CTAB. The increase in lambda(max) of BSA with the addition of CB in the reversed micellar phase might indicate the decrease in the hydrophobic microenvironment of the Trp residue of BSA, contrary to those microenvironments in the absence of CB. PMID- 11732714 TI - Isolation of a bacterium from mangrove soil for degradation of sea sludge. AB - Sea sludge, which is sediment of fish excrement and sewage on the sea bottom, continues to be a serious environmental problem. It has the potential to cause eutrophication and red tide, resulting in the death of shellfish and leading to an offensive odor. Soil taken from a mangrove swamp was added to sea sludge, which promoted an initial fermentation of the sludge components. This article reports on the isolation of a bacterium from mangrove soil that is involved in that fermentation. Three bacteria were isolated on a marine agar plate after incubating for 12 h at 60 degrees C. One of these bacteria fermented sea sludge. 16S rDNA of this bacterium was sequenced, and it had a high homology with that of Bacillus fumarioli LMG17489 (AJ250056). PMID- 11732715 TI - Synthesis and characterization of a novel extracellular polysaccharide by Rhodotorula glutinis. AB - The aim of this work was to characterize an exopolysaccharide by Rhodotorula glutinis KCTC 7989 and to investigate the effect of the culture conditions on the production of this polymer. The extracellular polysaccharide (EPS) produced from this strain was a novel acidic heteropolysaccharide composed of neutral sugars (85%) and uronic acid (15%). The neutral sugar composition was identified by gas chromatography as mannose, fucose, glucose, and galactose in a 6.7:0.2:0.1:0.1 ratio. The molecular weight of purified EPS was estimated to be 1.0-3.8 x 10(5) Dalton, and the distribution of the molecular weight was very homogeneous (polydispersity index = 1.32). The EPS solution showed a characteristic of pseudoplastic non-Newtonian fluid at a concentration >2.0% in distilled water. The maximum EPS production was obtained when the strain was grown on glucose (30 g/L). Ammonium sulfate was the best suitable nitrogen source for EPS production. The highest yield of EPS was obtained at a carbon to nitrogen ratio of 15. The EPS synthesis was activated at the acidic range of pH 3.0-5.0 and increased when the pH of the culture broth decreased naturally to <2.0 during the fermentation. When the yeast was grown on glucose (30 g/L) and ammonium sulfate (2 g/L) at 22 degrees C at an initial pH of 4.0, EPS production was maximized (4.0 g/L), and the glucose-based production yield coefficient and carbon-based production yield coefficient were 0.30 g of EPS/g of glucose and 0.34 g (carbon of EPS)/g (carbon of glucose), respectively. PMID- 11732716 TI - Manufacture of xylose-based fermentation media from corncobs by posthydrolysis of autohydrolysis liquors. AB - Milled corncob samples were mixed with water and heated to obtain a liquid phase containing oligosaccharides, sugars, and acetic acid as main reaction products (autohydrolysis reaction). To hydrolyze the sugar oligomers to the correspondent monomers, sulfuric acid was added to the autohydrolysis liquors to reach 0.5-2 wt% of solution, and the reaction media were heated at 101.5-135 degrees C. With this operational procedure, sugar solutions suitable as fermentation media (containing xylose as the major component) were obtained. The kinetics of the posthydrolysis step was characterized on the basis of experimental data concerning the time courses of the concentrations of xylooligosaccharides, xylose, furfural, and acetic acid. The concentrations of other reaction byproducts (glucose or arabinose) were also measured. PMID- 11732717 TI - Characterization of yeast strains for wine production: effect of fermentation variables on quality of wine produced. AB - Sixteen yeast strains isolated from grapefruit (Citrus paradis), orange (Citrus sinensis) and pineapple (Ananas comosus) were characterized using standard microbiological procedures. The species were identified as Saccharomyces uvarum, S. cerevisiae, S. carlbergensis, and S. ellipsoideus. Their abilities for wine production were tested by using sugar and ethanol tolerance tests. The best biochemically active strain, S. ellipsoideus, was used along with commercially available baker's yeast (S. cerevisiae) to produce wine from grapefruit, orange, and pineapple juices. After fermentation for 14 d with S. cerevisiae and 21 d with S. ellipsoideus, wines produced were compared with Baron de Valls (standard). The highest (10.47% [v/v]) and lowest (7.68% [v/v]) alcohol concentrations with corresponding residual sugar concentrations of 1.88% (w/v) and 7.7% (w/v) were produced from orange after fermentation with S. cerevisiae and S. ellipsoideus, respectively. S. ellipsoideus was found to be the best yeast strain producing wine with the highest acceptable score of 7.41 from orange. The study revealed the possibility of producing wine from our locally available fruits using simple, cheap, and adaptable technology with biochemically characterized yeast strains. PMID- 11732718 TI - Production of native and recombinant lipases by Candida rugosa: a review. AB - The yeast Candida rugosa produces multiple lipase isoenzymes sharing high sequence homology but with some differences in their catalytic properties. The regulation of C. rugosa lipase (CRL) synthesis and secretion in C. rugosa obeys a complex pattern. Fermentation processes for both wild-type and mutant C. rugosa strains are available for lipase production. Native CRL preparations have been extensively used for biotransformations. However, their inherent mixture of isoforms with variable profiles complicates interpretation and brings into question the reproducibility achieved between preparations. Although heterologous CRLs gene expression had been hampered owing to a nonuniversal codon usage, recent advances have made heterologous CRLs available. This will expand and improve the industrial utility of CRLs even further. The purpose of this review is to provide a summary of the recent advances on the production of native and recombinant lipases by C. rugosa. PMID- 11732719 TI - Cutaneous wound healing: an update. AB - Our understanding of wound-healing mechanisms has progressed over the past decade. Wound healing is traditionally divided into three phases--the inflammatory phase, the proliferation phase, and the remodeling phase--and involves a well-orchestrated interaction among blood vessels (platelets, macrophages, neutrophils, endothelial cells, and smooth muscle cells), epidermis (keratinocytes, melanocytes, and Langerhans cells), adnexal structures (outer root sheath cells and hair dermal papilla cells), dermis (fibroblasts and myofibroblasts), nervous system (neurons), and subcutaneous fatty layers (adipocytes). We review recent discoveries of basic and clinical aspects of wound healing including several revolutions that occurred in wound management: occlusive dressing therapy, use of living skin equivalents, and topical administration of growth factors. As we previously proposed, the use of tissue substitutes and autologous epidermal sheets led to a new concept of skin grafting through the keratinocyte activation phase in the graft healing mechanism. In this review, we also discuss a representative patient who presented with plantar wounds caused by calcaneal osteomyelitis and healed by the coverage of epidermal grafting. PMID- 11732720 TI - Comparative efficacy of ketoconazole and fluconazole in the treatment of pityriasis versicolor: a one year follow-up study. AB - Pityriasis versicolor can be treated by a single or multiple dosage regime of ketoconazole as well as by fluconazole. The therapeutic efficacy of these two drugs has not been compared. One hundred and eighty patients with moderate to extensive pityriasis versicolor confirmed by KOH and Wood's lamp examination were randomly assigned to one of the four oral antifungal regimes: Ketoconazole 400 mg single dose (Category I), Ketoconazole 200 mg daily for 10 days (Category II), Fluconazole 400 mg single dose (Category III) or Fluconazole 150 mg per week for 4 weeks (Category IV). Follow up was done at 2 and 4 weeks and then at 3, 6 and 12 months after the treatment in each group. KOH and Wood's lamp examinations were repeated each time. After four weeks of treatment, clinical cure was observed in 66.6% (Category I), 73.3% (Category II), 80% (Category III) and 59.9% (Category IV) of patients. Mycological cure after four weeks of treatment was observed in 53.3% (Category I), 73.3% (Category II), 82.2% (Category III) and 64.4% (Category IV) of patients. After twelve months of follow-up, maximum relapses were observed with Category I. No relapse was seen in Category III patients. The time period of relapse varied from three to ten months. In conclusion, single dose 400 mg oral fluconazole provided the best clinical as well as mycological cure rate with no relapse during twelve months of follow-up. PMID- 11732721 TI - Repigmentation after autologous miniature punch grafting in segmental vitiligo in North Indian patients. AB - Segmental vitiligo is rather refractory to medical treatment. The paucity of literature on the results of autologous miniature punch grafting in segmental vitiligo in dark-skinned patients led us to conduct such a study in 15 dark skinned patients of North Indian origin to observe the repigmentation response to this technique at different sites. The age range was 16-42 years (mean 23.08 years). There were four males and eleven females (M:F ratio being 1:2.75) who had the disease for 3-15 years. After autologous miniature punch grafting, a remarkable repigmentation was obtained in 12 (80%) cases with 90% to 100% improvement. About 40-60% repigmentation was observed within 3-6 months in the majority (66.6%) of cases and near total to total repigmentation was observed within 2 years. The maximum pigment spread was observed over the face (cheeks) and neck, and the minimum pigment spread was observed over the left arm and right upper eyelid. Complications were minimal. The present study demonstrates that autologous miniature punch grafting is a safe and easy technique, that produces good cosmetic results in the unsightly depigmenting lesions of segmental vitiligo, especially on the face and exposed areas, even in dark-skinned races if patients are selected carefully. This can be of great benefit to patients who are psychologically disturbed by their disease. PMID- 11732722 TI - The occurrence of various collagen diseases in one family: a sister with ISSc, PBC, APS, and SS and a brother with systemic lupus erythematosus. AB - We encountered siblings who had collagen diseases and related symptoms. Case 1 was a 53-year-old woman who had limited cutaneous systemic sclerosis (ISSc) associated with primary biliary cirrhosis (PBC), antiphospholipid antibody syndrome (APS), and subclinical Sjogren's syndrome (SS). Case 2 was a 48-year-old man, her younger brother, with systemic lupus erythematosus (SLE) that developed at 32 years of age. Investigation of their family revealed that their mother had Raynaud's phenomenon, arthritis, and subclinical Sjogren's syndrome, and that another younger brother of Cases 1 and 2 had Raynaud's phenomenon and general fatigue. HLA analysis revealed that the sister and brother had some identical HLA antigens in common, including A2, A33 (19), B67, B44 (12), Cw7, DR2, DR6, DR52, and DQ1. The sister, brother and their mother had common HLA antigens including A2, B67, Cw7, DR2, and DQ1. Although Cases 1 and 2 shared the same HLA system, they presented different phenotypes of collagen disease. PMID- 11732723 TI - Contact leukoderma of the scalp or an unusual variant of vitiligo? AB - Contact leukoderma due to hair dyes is strongly suspected in patients presenting with depigmented patches sharply localized to the scalp. We describe three patients with a striking pattern of depigmentation, which stopped abruptly at the hair margins. However, no definite correlation with the use of hair dyes could be made in any of them. Our cases represent an unusual pattern of vitiligo of the scalp, which seems to have been previously attributed mainly to hair colorants. Careful examination of the lesions on the scalp may reveal more cases with this pattern. PMID- 11732724 TI - Staphylococcal scalded skin syndrome developing during burn treatment. AB - We report a case of staphylococcal scalded skin syndrome (SSSS) in a 5-year-old girl who had been hospitalized for burn treatment. When she developed an upper respiratory tract infection, she manifested extensive erythema and exfoliation. There was a purulent discharge from the ulcer caused by the burn, and exfoliation was observed in the surrounding area. Based on clinical symptoms and laboratory data, SSSS was diagnosed and treated with antibiotics, resulting in subsidence. Staphylococcus aureus was isolated from the posterior nasopharynx and the skin erosion and was proved to produce exfoliative toxin A (ETA). Infection from an asymptomatic carrier of an ET-producing strain was suspected, but we failed to identify the origin, in spite of a thorough inspection of the mother, nurses, and physician. SSSS is occasionally reported as a hospital-acquired infection. We should study the frequency of asymptomatic carriers of ET-producing strains so that we can formulate strategies to prevent such infections. PMID- 11732725 TI - An Indian case of keratoderma hereditarium mutilans (Vohwinkel's syndrome) associated with ichthyosiform dermatosis. AB - Keratoderma hereditarium mutilans (KHM) or Vohwinkel's syndrome (VS) is usually inherited as an autosomal dominant disease, but a recessive type has occasionally been described. We report a 19-year-old male born to consanguineous parents, who presented with KHM associated with generalized ichthyosiform dermatosis. On examination, his generalized ichthyosiform dermatosis was accompanied by severe 'honey comb' like palmoplantar keratoderma and marked hyperhidrosis. Bilateral fifth toes showed complete constricting bands just distal to the metatarsophalangeal joints leading to pseudoainhum. The histopathologic examination of a biopsy from the sole showed hyperkeratosis, marked parakeratosis, hypergranulosis, acanthosis, and a few mononuclear cells in the papillary dermis, consistent with the diagnosis of KHM. Several hereditary disorders may show varying degrees of hyperkeratosis and constriction of digits; their differentiating features are discussed. This case is reported for its rarity and interesting associated findings. PMID- 11732726 TI - A case of pedunculated malignant melanoma of the vaginal mucosa. AB - We report an 84-year-old Japanese woman who presented with a pedunculated malignant melanoma of the vaginal mucosa. Mucosal melanoma is believed to be more common in Japan than other countries, but such tumors of the vulvovaginal region are quite unusual. In our patient, three tumors were connected by a narrow pedicle. Three black tumors measuring 5-10 mm in diameter with a common pedicle were seen on the vaginal mucosa at five o'clock from the cervix. The tumors were hanging from the narrow pedicle. On histologic examination, they were diagnosed as malignant melanoma. Resection was done with a distal margin of 3 cm from the tumors and a margin of 1 cm from the cervix. The patient has had no evidence of local recurrence or distant metastasis. In our patient, the three main tumors had a common pedicle, which seems to be a unique finding. Since pedunculated malignant melanomas are rare, making a clinical diagnosis is difficult. Although pedunculated melanomas are recognized as having a high malignant potential because these lesions are generally thick, a relatively good outcome is sometimes reported. In our patient, there was no tumor infiltration into the dermis of the pedicle, and this may be one reason for the good outcome at present. There has been no previous report of a mucosal melanoma consisting of three tumors like those in the present patient. PMID- 11732727 TI - Papular elastorrhexis in childhood improved by intralesional injections of triamcinolone. AB - Papular elastorrhexis is a rare disease developing asymptomatic skin-colored small papules in adolescence with histopathological loss of elastic fibers. There has been no established treatment for this disease. A 4-year-old Korean boy had multiple, hard, whitish papules on his chest and back for one year. Histopathologic examination revealed focal loss of elastic fibers in the dermis, and X-ray examination showed no bony abnormalities. His skin lesions were improved by intralesional injections of triamcinolone but recurred after four months. PMID- 11732728 TI - A case of cutaneous bronchogenic cyst over the left scapula. AB - Bronchogenic cyst is noted shortly after birth or in early childhood and usually presents as a swelling or draining sinus in the presternal area. Its origin and pathogenesis can be explained as a developmental anomaly of the tracheobronchial buds from the primitive foregut. The patient was a 4-year-old boy with a child fist-sized soft mass over his left scapula, which had been detected at birth and had been gradually growing. Grossly, it appeared to be a simple cyst with clear mucoid fluid. Histopathological study demonstrated a unilocular cyst composed of ciliated pseudostratified columnar epithelia, interspersed goblet cells, smooth muscles, and mucous glands on the cyst wall, which are features compatible with cutaneous bronchogenic cyst. PMID- 11732729 TI - A case of generalized morphea and polymyositis accompanied by sick sinus syndrome. PMID- 11732731 TI - Propolis-induced erythroderma. PMID- 11732730 TI - Calcinosis cutis in a patient with primary hyperoxaluria due to hepatic enzyme deficiency. PMID- 11732732 TI - Profound skin infection with bone involvement due to Nocardia asteroides in a patient with myelodysplastic syndrome. PMID- 11732733 TI - Cutaneous findings in HIV-1-positive patients in Thailand. PMID- 11732734 TI - Nuclear and cellular volumetric alterations in oral lichen planus and lichenoid lesions: a histomorphometric study. AB - There is presently no line of distinction between oral lichen planus and other oral lichenoid lesions. The aim of this study is to determine using histomorphometry, the differences between these lesions. Paraffin sections from 7 normal buccal epithelium, 19 oral lichen planus (LP), 14 oral lichenoid lesions (LL) and 7 discoid lupus erythematosus-like lesions (DLE-ll) were selected. The nuclear volume (V(N)) and cellular-volume (V(CELL)) of the epithelium were assessed using an image analyser. The V(N) and V(CELL), derived for both basal and spinal strata in LP and DLE-ll were 2.3 times more than that of normal tissues. There was a significant difference between LP and LL (P < 0.005) and between LL and DLE-ll (P < 0.001), but not between LP and DLE-ll. In conclusion, there appears to be a difference between LP, LL and DLE-ll and V(N) and V(CELL) may serve as potential discriminators between these groups of lesions. PMID- 11732735 TI - Inhibition of hemolysis by antibody against the Porphyromonas gingivalis 130-kDa hemagglutinin domain. AB - Porphyromonas gingivalis is a gram-negative anaerobic bacterial species implicated as an important pathogen in the development of adult periodontitis. Hemagglutinin may mediate the adsorption and invasion of bacteria into host cells. Furthermore, the hemagglutinin plays a role in the agglutinate and lyse erythrocytes intake of heme which is an absolute requirement for this bacterial growth. We previously cloned the gene encoding the 130-kDa hemagglutinin protein domain (130-kDa HMGD) and identified the functional motifs of agglutination of erythrocytes. Bacterial cell attachment to erythrocytes is an important initial step in the expression of hemolytic activity. In this study, we highly purified recombinant r130-kDa HMGD and prepared the specific antiserum. Further, the effect of the antibody on the hemolytic activity of P. gingivalis cells was examined. The polyclonal antibody recognized 43,49-kDa major bands in P. gingivalis cells and r130-kDa HMGD, and significantly inhibited the hemagglutinating and hemolytic activities of P. gingivalis cells. The findings suggest that the antibody may be useful in the development of the passive immunization against periodontal diseases caused by P. gingivalis infection. PMID- 11732736 TI - Papillary squamous cell carcinoma of the oral mucosa: immunohistochemical comparison with other carcinomas of oral mucosal origin. AB - Papillary squamous cell carcinoma (PSCC) is a poorly described variant of squamous cell carcinoma, and may be confused with verrucous carcinoma of the head and neck. To add to existing knowledge of this rare tumor, we describe two cases of PSCC arising in the oral mucosa. The lesions were composed of exophytic proliferation of atypical to overtly malignant cells similar to those of conventional squamous cell carcinoma, and invasion into the superficial region of the underlying fibrous tissue was seen in the form of islands and cords of malignant cells. Immunohistochemical assessment of cellular proliferative activity showed a significantly high mean percentage of Ki-67 expression in comparison with verrucous carcinoma, but there was no significant difference of Ki-67 expression among PSCC, conventional squamous cell carcinoma and microinvasive squamous cell carcinoma. These results suggest that the biological behavior of PSCC is analogous to that of SCC. PMID- 11732737 TI - Interleukin-1 beta levels in gingival crevicular fluid in type 2 diabetes mellitus and adult periodontitis. AB - Interleukin-1 beta (IL-1beta) is a potent bone-resorptive cytokine that also mediates soft-tissue destruction by stimulating prostaglandin production and inducing collagenase and other protease activity. The literature suggests that this substance may be an important mediator of attachment loss in human periodontitis, and indicates that IL-1beta may be useful for locating sites of periodontal disease activity. There is some evidence that IL-1beta is produced by cells of the periodontium, and that it can be detected in gingival crevicular fluid (GCF). Many factors are known to contribute to the destruction of periodontal tissue. One of the most important is immune deficiency in diabetes. The aim of this study was to measure and compare the concentration of IL-1beta in the GCF of patients with non-insulin-dependent diabetes mellitus (Type 2 DM), otherwise healthy adults with periodontitis, and individuals with no periodontal disease in order to assess whether diabetes alters IL-1beta levels. We also examined relationships between GCF levels and the clinical parameters of pocket depth, plaque index, and bleeding index in each group. Seventeen patients with Type 2 DM, 17 adult periodontitis patients (AP), and 17 healthy controls were selected. The levels of IL-1beta in the GCF were quantified by ELISA. The mean IL 1beta concentrations in the Type 2 DM, AP, and control groups were 200.1 +/- 65.34 pg/microl, 131.35 +/- 67.66 pg/microl, and 80.0 +/- 36.08 pg/microl, respectively. The levels in the diabetic patients were significantly higher than those in the AP and control subjects. There were no significant correlations between IL-1beta level and any of the clinical data parameters for each group. We believe that the macrophages may over produce IL-beta in Type 2 DM and increased IL-1beta levels in diabetic patients could be linked to altered immune function. PMID- 11732738 TI - Gene expression and immunohistochemical localization of decorin and biglycan in association with early bone formation in the developing mandible. AB - We investigated the expression of the small proteoglycans, decorin and biglycan, which are associated with osteoblast differentiation, and how this relates to the expression of osteocalcin and bone sialoprotein (BSP) early in the formation of bone in the rat mandible by immunohistochemistry and in situ hybridization. The mandibles of rat fetuses were collected on embryonic days 14 (E14) to E18. In situ hybridization showed that gene expression of decorin, biglycan, osteocalcin and BSP was not apparent in the developing mandible at E 14, but was expressed by newly differentiated osteoblasts at E15. The expression of these mRNAs increased linearly as the number of osteoblasts increased in specimens from E16 to E18. Immunohistochemistry showed that newly differentiated osteoblasts expressed biglycan moderately, decorin weakly, and osteocalcin and BSP faintly. The unmineralized bone matrices among the osteoblasts showed prominent staining for decorin, weak staining for osteocalcin and BSP, and very weak staining for biglycan. When the intercellular matrix was mineralized at E16, the mineralized bone matrix showed more prominent staining for osteocalcin and BSP, but lacked staining for decorin and biglycan. The same staining profile was observed during the subsequent phases of bone formation at E17 and E18. These results indicate that decorin, biglycan, osteocalcin and BSP are expressed at the gene and protein level by newly differentiated osteoblasts before the onset of matrix mineralization and that they could play a role in the earliest stages of bone formation. Negative proteoglycan staining in the mineralized bone matrix suggests that a loss of, or a sharp decrease in proteoglycans occurs concomitant with bone matrix mineralization. PMID- 11732739 TI - A pilot study of measurement of the frequency of sounds emitted by high-speed dental air turbines. AB - Since the development and use of the high-speed dental air turbine some 45 years ago, concern has been expressed in the literature about a possible cause and effect relationship between use of the drill and hearing loss in dentists. The hearing threshold in humans varies with the frequency of sound. It is well known that dentists experience gradual hearing loss during their working life. The aim of this study was to measure the frequency of sounds emitted by high-speed dental air turbines under different working conditions. Five high-speed dental air turbines were used (2 x Trend TC-80 BC W&H Dentalwerk, Austria, 2 x Black Pearl Eco Bien-air, Switzerland, 1 x Trend TC-80 BC W&H Dentalwerk, Austria. Each turbine was tested under 8 different working conditions: under free working conditions the turbines were tested without burs, with fissure burs, with flare burs, with round burs and with inverted cone burs; under operation they were tested with fissure burs by application to a 3 x 3 x 10 mm amalgam block surface, a 3 x 3 x 10 mm composite block surface, and the occlusal surface of an extracted molar tooth. Forty sound recordings were made in total using a computer with a microphone (Shure 16 LC) located 30 cm away from the samples, at 10-s intervals using a mixer. Frequency analysis was done by a Cool Edit Pro 1.2 computer program. Data were analyzed by multi-variate analysis with the S.P.S.S 9.05 software program. The average measurement was 6860 Hz. According to the statistical analysis there was no significant difference in the frequencies recorded under different working conditions. There was also no significant difference among the different high-speed dental air turbines at alpha = 0,05, P > alpha /2 levels. These results indicate that under any working conditions, high speed dental air turbines emit frequencies which can cause hearing loss. PMID- 11732740 TI - Color stability of different denture teeth materials: an in vitro study. AB - The aim of this in vitro study was to compare the color stability of commercially available porcelain, reinforced acrylic, and conventional acrylic denture teeth materials used in removable prostheses. Two brands of porcelain (Unilux-Enta Lactona-Holland and Vivoperl-Ivoclar-Liechtenstein), 2 brands of reinforced acrylic (Optodent-Bayer-Germany and Ivolek-Ivoclar-Liechtenstein), and 2 brands of conventional acrylic (Isodent-Guney Dis Deposu-Turkey and Samed-Turkey), were made, for a total of 6 different denture teeth groups. Denture teeth were subjected to 3 staining solutions (filtered coffee, tea, and cola) and distilled water. From each group of denture teeth, 4 sets of maxillary anterior denture teeth were immersed in each of the 4 solutions. The color values of denture teeth were measured colorimetrically with the Gardner XL 20 Tristimulus Colorimeter (Gardner Lab. Inc., Bethesda, Maryland, USA). Color changes were characterized in the CIEL*a*b* color space. Color change values were determined after 1 day, 1 week, 2 weeks, and 4 weeks. The color difference values were calculated and then evaluated by two-way ANOVA statistically. The filtered coffee solution was found to be more chromogenic than the other 2 staining solutions, while porcelain denture teeth materials were more color stable. Assuming the color change of deltaE* < 1.0 as a discernible limit and deltaE* = 3.3 as an acceptable value, the filtered coffee, tea, and cola had slight staining effects on all 6 groups of denture teeth. PMID- 11732741 TI - Enhancement of plasminogen activator activity stimulated by LPS in gingival fibroblasts of individuals with Down syndrome. AB - Individuals with Down syndrome (DS) have a high prevalence of periodontal disease, which develops early and progressed rapidly and extensively, in comparison with healthy controls. The severe periodontal disease in individuals with DS has been considered to result from abnormal factors in their host responses. The mechanisms involved in the periodontal inflammatory processes in individuals with DS are not fully understood. Plasminogen activators (PA) are serine proteases that are well known for their part in the initiation of the fibrinolytic cascade leading to the generation of plasmin in periodontal homeostasis, including fibrinolysis and connective tissue remodeling. The PA plasmin system affects the progression of periodontal disease. In the present study, we examined the effects of the levels of PA activity stimulated with lipopolysaccharide (LPS) in the gingival fibroblasts from donors with DS (DGF). The levels of PA activity without LPS were low in the DGFs, the same as that in the gingival fibroblasts from donors of healthy controls (NDGF). In contrast, the levels of PA activity with LPS in DGFs were significantly higher than that in the NDGFs. These results suggested that PA plays an important role in inducing extensive and rapid inflammation in the periodontal disease in individuals with DS. PMID- 11732742 TI - An unusual staining of the tooth roots: a case report with histological and micro analytical studies. AB - The discoloration of tooth roots is rare. We report here a 22-year-old Japanese woman with blackish-brown staining of the roots of the upper and lower third molars. Staining was found in the dentin and cementum. Electron probe X-ray microanalysis showed no significant difference in the composing elements between the stained tooth root and control tooth. Fluorescent bands coincided with staining in the dentin of the root and cementum along the incremental lines under confocal laser-scanning microscope. PMID- 11732743 TI - Oral nodular fasciitis: a case report. AB - Nodular fasciitis of the cheek is an extremely rare lesion of the oral cavity. It should be considered in the differential diagnosis of swellings in the oral mucosa. We describe a case of nodular fasciitis and discuss the difficulties of histological and clinical diagnosis of this condition. PMID- 11732744 TI - Alstrom syndrome: a case report. AB - Alstrom syndrome is a rare disorder characterized by early obesity, loss of central vision, diabetes mellitus, hearing loss and short stature. Previous studies, have reported no information regarding oral findings. This article describes oral findings in two cases of Alstrom syndrome. In both cases, gingivitis was present and also light yellow-brown discolored enamel bands were observed on the anterior teeth. This staining may have resulted from discoloration of the preexisting slight band-like enamel hypoplasia. The gingiva was examined histologically by light and transmission electron microscopy. Irregular thickness of the basal lamina and delamination of the myelin sheath were detected by transmission electron microscopy. There is no information about pathological odontogenesis in Alstrom syndrome in previous reports. Oral present findings may contribute further information about the clinical manifestations of Alstrom syndrome. PMID- 11732745 TI - Growth factor release from tissue engineering scaffolds. AB - Synthetic scaffold materials are used in tissue engineering for a variety of applications, including physical supports for the creation of functional tissues, protective gels to aid in wound healing and to encapsulate cells for localized hormone-delivery therapies. In order to encourage successful tissue growth, these scaffold materials must incorporate vital growth factors that are released to control their development. A major challenge lies in the requirement for these growth factor delivery mechanisms to mimic the in-vivo release profiles of factors produced during natural tissue morphogenesis or repair. This review highlights some of the major strategies for creating scaffold constructs reported thus far, along with the approaches taken to incorporate growth factors within the materials and the benefits of combining tissue engineering and drug delivery expertise. PMID- 11732746 TI - Comparison of the lymphatic transport of a lipophilic drug from vehicles containing alpha-tocopherol and/or triglycerides in rats. AB - The applicability of alpha-tocopherol as a lymphotropic carrier for a highly lipophilic drug has been evaluated. Transport to the intestinal lymph of the highly lipophilic model drug, Lu28-179, in rats after administration to the stomach in an alpha-tocopherol emulsion was compared with lymphatic transport after administration of a sesame oil emulsion and an alpha-tocopherol/sesame oil emulsion. Lymphatic transport of the triglycerides and of alpha-tocopherol was determined. A conscious rat model was used, and the mesenteric lymph was collected. There was no significant difference between the cumulative masses of triglyceride from the two emulsions containing triglyceride 24 h after administration. Administration of an alpha-tocopherol emulsion seemed to induce mobilization of endogenous triglyceride. The lymphatic transport of alpha tocopherol was less than 1 mg 24 h after administration of both emulsions containing alpha-tocopherol. The absorption of Lu28-179 from the alpha-tocopherol emulsion was very low, with a lymphatic recovery of 0.05%. When administered in an alpha-tocopherol/sesame oil emulsion, the recovery of Lu28-179 increased sevenfold to 0.35%. However, after administration of Lu28-179 in a sesame oil emulsion, the lymphatic recovery increased a further 13-fold to 4.5%. In conclusion, the study showed that alpha-tocopherol did not promote lymphatic absorption of Lu28-179 and thus was not a good lymphotropic carrier, as compared with sesame oil. Alpha-tocopherol in combination with sesame oil was not a good lymphotropic carrier either. The non-absorbed alpha-tocopherol fraction in the intestine might be able to prevent the absorption of Lu28-179. PMID- 11732747 TI - In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies. AB - The aim of this study was to establish a collagen matrix-based nasal primary culture system for drug delivery studies. Nasal epithelial cells were cultured on derivatised (Cellagen membrane CD-24), polymerised (Vitrogen gel) and fibrillar (Vitrogen film) collagen substrata. Cell morphology was assessed by microscopy. The cells were further characterised by measurement of ciliary beat frequency (CBF), transepithelial resistance (TER), permeation of sodium fluorescein, mitochondrial dehydrogenase (MDH) activity and lactate dehydrogenase (LDH) release upon cell exposure to sodium tauro-24, 25 dihydrofusidate (STDHF). Among the three collagen substrata investigated, the best epithelial differentiated phenotype (monolayer with columnar/cuboidal morphology) occurred in cells grown on Cellagen membrane CD-24 between day 4 and day 11. Cell culture reproducibility was better with Cellagen membrane CD-24 (90%) in comparison with Vitrogen gel (70%) and Vitrogen film (< 10%). TER was higher in cells grown on Vitrogen gel than on Cellagen membrane CD-24 and Vitrogen film. The apparent permeability coefficient (Papp x 10(-7)cm s(-1)) of sodium fluorescein in these conditions was 0.45+/-0.08 (Vitrogen gel) and 1.91+/-0.00 (Cellagen membrane CD-24). Except for LDH release, CBF and cell viability were comparable for all the substrata. Based on MDH activity, LDH release, CBF, TER and permeation studies, Cellagen membrane CD-24- and Vitrogen gel-based cells were concluded to be functionally suitable for in-vitro nasal drug studies. Vitrogen film-based cultures may be limited to metabolism and cilio-toxicity studies. PMID- 11732748 TI - Ursodeoxycholic acid action on the transport function of the small intestine in normal and cystic fibrosis mice. AB - Ursodeoxycholic acid possesses choleretic and cytoprotective properties and in cystic fibrosis (CF) it is used to treat the hepatobiliary symptoms of the disease. This study investigated the effects of this bile acid on the transport function of the small intestine in normal and CF mice. The effects of ursodeoxycholic acid were monitored as changes in short-circuit current (SCC) in stripped sheets of small intestine from normal (Swiss MF1) and transgenic CF (Cftr(tm2Cam)) mice. In ileal sheets from Swiss MF1 mice, mucosal ursodeoxycholic acid caused a biphasic increase in SCC. The first phase was reduced by lowering the mucosal Na+ concentration, while the second phase was inhibited by (Cl-)-free conditions, serosal furosemide or mucosal diphenylamine-2-carboxylic acid (DPC), suggesting an initial Na+-dependent bile acid absorption followed by a stimulation of electrogenic Cl- secretion. Serosal application of ursodeoxycholic acid to the ileum and mucosal or serosal application to the mid-intestine and jejunum elicited a secretory response only. Secretion was Ca2+-dependent, but did not involve neural mechanisms. Mucosal mast cells, histamine and serotonin (5-HT) were implicated in the secretory response. Responses in tissues from transgenic wild-type mice were similar to those obtained with Swiss MF1 mice, but the secretory response to mucosal or serosal application of the bile acid was impaired in CF tissues. In ilea from CF mice the initial absorptive phase of the response to mucosal ursodeoxycholic acid was still observed. It is concluded that ursodeoxycholic acid induces secretion throughout the murine small intestine by a mechanism that involves degranulation of mucosal mast cells. In the ileum Na+ dependent absorption can also be detected. The secretory response is defective in CF intestine, but the absorptive effect is still present. PMID- 11732749 TI - In-vitro activity and tissue distribution of new fluorinated meso tetrahydroxyphenylporphyrin photosensitizers. AB - Tetra(hydroxyphenyl)porphyrins started to attract interest as potential photosensitizers for photodynamic therapy in the early eighties. Subsequently, a number of derivatives of these compounds have been studied. In 1997 we reported the synthesis of the fluorinated derivatives 5,10,15,20-tetrakis(2-fluoro-3 hydroxyphenyl)porphyrin (8), 5,10,15,20-tetrakis(2,4-difluoro-3 hydroxyphenyl)porphyrin (9), and 5,10,15,20-tetrakis(3,5-difluoro-4 hydroxyphenyl)porphyrin (10). We have measured their biological activity, using the MTT test, against cancer cell cultures in-vitro. The test showed that these compounds were as potent as 5,10,15,20-tetrakis(3-hydroxyphenyl)chlorin (5), one of the leading photosensitizers in photodynamic therapy. The highest photoactivity was shown by the meta-hydroxy compounds 8 and 9. The para-compound showed high toxicity in the dark. Distribution of these compounds between normal and cancer tissue was studied using 19F NMR spectroscopy. The highest cancer tissue localization was also shown by the meta-hydroxy compounds 8 and 9. The para compound showed poor localization in tumour tissue. This study has shown that 19F NMR spectroscopy can be used to estimate the tissue distribution of fluorinated tetrahydroxyphenylporphyrins in-vivo. PMID- 11732750 TI - Effects of cholesterol and model transmembrane proteins on drug partitioning into lipid bilayers as analysed by immobilized-liposome chromatography. AB - We have analysed how cholesterol and transmembrane proteins in phospholipid bilayers modulate drug partitioning into the bilayers. For this purpose we determined the chromatographic retention of drugs on liposomes or proteoliposomes entrapped in gel beads. The drug retention per phospholipid amount (the capacity factor Ks) reflects the drug partitioning. Cholesterol in the bilayers decreased the Ks value and hence the partitioning into the membrane in proportion to the cholesterol fraction. On average this cholesterol effect decreased with increasing temperature. Model transmembrane proteins, the glucose transporter GLUT1 and bacteriorhodopsin, interacted electrostatically with charged drugs to increase or decrease the drug partitioning into the bilayers. Bacteriorhodopsin proteoliposomes containing cholesterol combined the effects of the protein and the cholesterol and approached the partitioning properties of red blood cell membranes. For positively charged drugs the correlation between calculated intestinal permeability and log Ks was fair for both liposomes and bacteriorhodopsin-cholesterol proteoliposomes. Detailed modeling of solute partitioning into biological membranes may require an extensive knowledge of their structures. PMID- 11732751 TI - Synthesis and in-vitro/in-vivo evaluation of orally administered entacapone prodrugs. AB - Entacapone is a new inhibitor of catechol-O-methyltransferase (COMT) that is used as an adjunct to L-dopa therapy in the treatment of Parkinson's disease. The bioavailability of orally administered entacapone is, however, relatively low (29 46%). In this study we have prepared more lipophilic acyl and acyloxyacyl esters, an acyloxy alkyl ether and an alkyloxycarbonyl ester of entacapone, and we have evaluated them as potential prodrugs to enhance the oral bioavailability of entacapone. All the derivatives fulfilled prodrug criteria and released entacapone in human serum in-vitro. The oral bioavailability of monopivaloyl (1a) and dipivaloyl (1b) esters of entacapone were investigated further in rats. The lipophilicity of 1b was high (log Papp 4.0 at pH 7.4) but its oral bioavailability was low (F = 0.6%), most probably due to its low aqueous solubility. The monopivaloyl ester of entacapone (1a) had a higher lipophilicity (log Papp 0.80) than entacapone (log Papp 0.18) at pH 7.4 while maintaining an aqueous solubility equal to entacapone. However, oral bioavailability was not increased when compared with the parent drug entacapone (F = 7.0% and 10.4%, respectively). PMID- 11732752 TI - Comparison of vaginal aminopeptidase enzymatic activities in various animals and in humans. AB - The specific enzymatic activity of four different aminopeptidases (aminopeptidase N, leucine aminopeptidase, aminopeptidase A and aminopeptidase B) in vaginal homogenates from rabbit, rat, guinea-pig, sheep and humans was compared. The purpose of the study was to find an appropriate animal model that can be used in degradation studies of protein and peptide drugs. Different substrates were used as the relative specific substrates for the determination of aminopeptidase enzymatic activity: 4-methoxy-2-naphthylamide of L-alanine for aminopeptidase N, 4-methoxy-2-naphthylamide of L-leucine for leucine aminopeptidase, 4-methoxy-2 naphthylamide of L-glutamic acid for aminopeptidase A and 4-methoxy-2 naphthylamide of L-arginine for aminopeptidase B. The vaginal aminopeptidase enzymatic activity of different species was determined spectrofluorometrically. The inhibition of aminopeptidase activity in the presence of bestatin and puromycin inhibitors was also investigated. The results showed the presence of aminopeptidase enzymatic activity in all vaginal homogenates in the order: sheep > guinea-pig > rabbit > or = human > or = rat. Based on the results of the hydrolysis and inhibition of the 4-methoxy-2-naphthylamide substrates, it was difficult to have an exact decision on the aminopeptidase type in the vaginal homogenates from the species studied. It was found that the aminopeptidase activity in rat, rabbit and humans was not statistically different. Therefore, we suggest that rats and rabbits could be used as model animals for vaginal enzymatic activity studies and for determination of the degradation of protein and peptide drugs in the vagina. PMID- 11732753 TI - Design, synthesis and biological testing of a novel series of anti-inflammatory drugs. AB - Many of the non-steroidal anti-inflammatory drugs (NSAIDs) currently marketed produce severe gastro-toxic side effects. The benefits of producing NSAIDs without these side effects are obvious, particularly for patients requiring long term therapy. The aim of this investigation was to produce novel NSAIDs, based on paracetamol, that exhibit little or no gastro-toxicity. The work covers design, synthesis and testing of 13 drug candidates. The analgesic and anti-inflammatory potencies of the drug candidates were measured using the mouse abdominal constriction assay and the carrageenan-induced rat paw oedema assay, respectively. The stomachs of the rats were examined post-mortem, to assess the gastro-toxicity of the drugs. Of the 13 compounds described herein, 11 were shown to possess analgesic activity at 2-10 times the potency of aspirin, while 8 demonstrated anti-inflammatory activity at 3-10 times the potency of aspirin. Significantly, all of the compounds showed very low gastro-toxicity when compared with aspirin. The results of this study indicate that it is possible to develop novel, potent NSAIDs based on the structure of paracetamol. These compounds have the advantage of demonstrating much lower gastro-toxicity than NSAIDs currently available. Drugs of this type may, in future, provide effective treatments for inflammatory disorders. PMID- 11732754 TI - Immunomodulating activity of CVT-E002, a proprietary extract from North American ginseng (Panax quinquefolium). AB - The activity of CVT-E002, an aqueous extract containing mainly oligosaccharides and polysaccharides from North American ginseng (Panax quinquefolium), as an immunobooster on murine spleen cells and peritoneal macrophages, was studied in vitro. CVT-E002 stimulated the proliferation of normal mouse spleen cells, of which the major responding subpopulation was identified as B lymphocytes. CVT E002 also activated peritoneal exudate macrophages leading to enhanced interleukin-1 (IL-1), interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF alpha) and nitric oxide (NO) production. In addition, CVT-E002 stimulated in-vivo immunoglobulin G (IgG) production in treated mice. These results identify some of the immunomodulating activities of CVT-E002 and suggest its use clinically for the modulation of immune responses. PMID- 11732755 TI - Protective effects of cannabinoid receptor agonists against cocaine and other convulsant-induced toxic behavioural symptoms. AB - Based on the previously reported co-localization and relationship between cannabinoid and dopamine receptors, the effects of cannabinoid receptor agonists against cocaine-induced toxic behavioural symptoms, including convulsive seizures, were examined in mice. The anticonvulsant effect of several cannabimimetics against seizures induced by other convulsants was also compared. The cannabinoid receptor agonists CP 55940 ((-)-cis-3-[2-hydroxy-4-(1,1 dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)-cyclohexanol) and WIN 55212-2 ((R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4 benzoxazin-6-yl]-1-naphthalenylmethanone), and the endogenous cannabinoid anandamide were co-administered intraperitoneally with cocaine (75 mg kg(-1)) or other convulsants such as bicuculline, methyl 6,7-dimethoxy-4-ethyl-beta carboline-carboxylate (DMCM), L-glutamic acid and N-methyl-D-aspartate (NMDA). CP 55940 (2.5 mg kg(-1)) and anandamide (15 mg kg(-1)) significantly antagonized cocaine-induced lethality, and CP 55940 and WIN 55212-2 (2.5 mg kg(-1)) significantly attenuated the severity of cocaine-induced convulsive seizures. Furthermore, ataxic hyperactivity, which was observed only in the cocaine-treated group of mice and could be evaluated by their activity counts, was also depressed in the groups of mice co-treated with each of the three cannabinoid agonists. However, none of these agonists protected against bicuculline- or DMCM-induced lethality or convulsive seizures. In contrast, all of the cannabinoid agonists, most notably anandamide, antagonized both L-glutamic acid (2 g kg(-1))- and NMDA (200 mg kg(-1))-induced convulsive seizures. These data support the previously reported close correlation between dopamine and cannabinoid receptors, and between cannabinoid agonists, especially anandamide, and glutamate (NMDA) receptors. Furthermore, these results suggest a potential therapeutic role for cannabinoid agonists against cocaine- and other-convulsant-induced toxicities. PMID- 11732756 TI - New insights into the mechanism of action of the anti-inflammatory triterpene lupeol. AB - The pentacyclic triterpene lupeol has been studied for its inhibitory effects on murine models of inflammation and peritoneal macrophage functions in-vitro. Lupeol (0.5 and 1 mg/ear) administered topically suppressed the mouse ear oedema induced by 12-0-tetradecanoyl-phorbol acetate (TPA), being less effective on ear oedema induced by arachidonic acid. Quantitation of the neutrophil specific marker myeloperoxidase demonstrated that its topical activity was associated with reduction in cell infiltration into inflamed tissues. When tested in-vitro, lupeol significantly reduced prostaglandin E2 (PGE2) production from A23187 stimulated macrophages, but failed to affect leukotriene C4 release. It was a weak inhibitor of nitrite release, but dose-dependently suppressed PGE2. Cytokine production (tumour necrosis factor-alpha and interleukin-1beta) was inhibited in the range 10-100 microM in lipopolysaccharide-treated macrophages. This study demonstrated that lupeol possessed anti-inflammatory activity which was likely to depend on its ability to prevent the production of some pro-inflammatory mediators. PMID- 11732757 TI - Effect of hyperprolactinaemia as induced by pituitary homografts under kidney capsule on gastric and duodenal ulcers in rats. AB - The effect of hyperprolactinaemia, induced by two or four pituitary homografts under the kidney capsule, on gastric and duodenal ulcers has been studied. The acute gastric ulcer models used were pylorus ligation, indometacin-induced and ethanol-induced gastric ulcers. Chronic gastric ulcers were induced using acetic acid and duodenal ulcers by mercaptamine hydrochloride. After pylorus ligation, there was an approximate 30-40% increase in gastric secretion, a significant increase in total acidity (P < 0.01) and in the ulcer index (P < 0.01) in rats bearing pituitary homografts under the kidney capsule when compared with the sham operated control. Hyperprolactinaemia did not affect the formation of ethanol induced gastric ulcers but showed a 40% reduction in the development of indometacin-induced gastric ulcers. It also produced a 20% increase in the ulcer index in acetic acid-induced chronic gastric ulcers and a 30% increase in ulcer area in mercaptamine-induced duodenal ulcers. Our results showed that hyperprolactinaemia induced gastric acid secretion and thereby aggravated gastric and duodenal ulcers in rats. Hyperprolactinaemia did not affect gastric cytoprotection. PMID- 11732758 TI - Response of MCF-7 human breast cancer cells to some binary mixtures of oestrogenic compounds in-vitro. AB - The effects of simultaneous administration of some binary mixtures of seven natural and synthetic oestrogenic substances (17beta-estradiol, estrone, bisphenol A, butylbenzyl phthalate, endosulfan, methoxychlor and pentachlorophenol) on the cellular proliferation of human breast cancer MCF-7 cells in-vitro (a modified E-screen assay) have been measured. To assess the presence or absence of interactions of the two agents, the data were analysed on the basis of a graphical method in which the types and extents of interactions were described by response-surface diagrams. Of the nine combinations of the agents examined, synergistic interaction was evident for the combination of 17beta-estradiol and bisphenol A, whereas the remaining eight combinations were weakly synergistic, additive and/or weakly antagonistic in the dose-range tested. PMID- 11732759 TI - Riluzole suppresses post-sigh, but not spontaneous apnoeas during sleep in rats. AB - We conducted this experiment to determine the role of glutamate in the mechanism of sleep apnoeas by administering riluzole, a glutamate release inhibitor, to freely moving rats in which sleep-related apnoeas are physiological phenomena. Adult Sprague-Dawley rats were implanted with electrodes for electroencephalogram (EEG) and electromyogram (EMG) recording to monitor sleep and were placed inside a single-chamber plethysmograph to monitor respiration. Sleep and respiration were recorded for 6 h following intraperitoneal administration of 0.5, 5.0 and 10.0 mg kg(-1) riluzole. Riluzole dose-dependently suppressed post-sigh apnoeas during rapid eye movement (REM) sleep but had no effect on sleep-related spontaneous apnoeas. The drug (5.0 and 10.0 mg kg(-1)) also dose-dependently reduced wakefulness and increased sleep. It appears that glutamate, an excitatory neurotransmitter, plays a role in the genesis of the post-sigh apnoeas during REM sleep. PMID- 11732760 TI - Synthesis and preliminary behavioural evaluation in mice of new 3-aryl-3-pyrrol-1 ylpropanamides, analogues of FGIN-1-27 and FGIN-1-43. AB - The 2-aryl-3-indoleacetamides FGIN-1-27 and FGIN-1-43 have already been characterized in-vitro as potent and specific ligands for the mitochondrial DBI receptor. This affinity was associated with psychotropic properties in several rodent behavioural tasks (in particular anxiolytic action) via enhancement of GABA transmission through neurosteroid production. The synthesis of new 3-aryl-3 pyrrol-1-ylpropanamides 1a-i, analogues of FGIN-1-27 and FGIN-1-43, is described in four steps starting from the corresponding arylaldehydes. Preliminary evaluation of these compounds in behavioural studies (spontaneous locomotor activity and anxiolytic activity) in mice was also undertaken. PMID- 11732761 TI - Hepatoprotective activity of Trichilia roka on carbon tetrachloride-induced liver damage in rats. AB - Trichilia roka Chiov. (Meliaceae) is a tree widely distributed in tropical Africa. It has been used in Mali folk medicine for the treatment of various illnesses. A decoction of the roots is taken as a remedy for colds and pneumonia, and it is used as a diuretic and in hepatic disorders. We have evaluated the hepatoprotective effects of a decoction of Trichilia roka root on CCl4-induced acute liver damage in rats. Treatment with the decoction showed a significant protective action made evident by its effect on the levels of glutamate oxalacetate transaminase and glutamate pyruvate transaminase in the serum, on the protein content and lipid peroxidation levels in the liver homogenate. Histopathological changes produced by CCl4, such as necrosis, fatty change, ballooning degeneration and inflammatory infiltration of lymphocytes around the central veins, were clearly recovered by the treatment with Trichilia root decoction. On fractionating this extract into diethyl ether-soluble and water soluble fractions, the activity was retained in the diethyl ether-soluble fraction. Moreover, the administration of decoction prevented a preferential deposition of collagen around the sinusoidal cell layer, which is responsible for the perisinusoidal fibrosis in the early stage of CCl4 damage. This study showed that treatment with Trichilia roka extracts or silymarin (as reference) appeared to enhance the recovery from CCl4-induced hepatotoxicity. The hepatoprotective properties of Trichilia roka may be correlated to polyphenol content of the decoction and its diethyl ether-soluble fraction. PMID- 11732762 TI - Long-term developmental outcome of asphyxiated term neonates. AB - Asphyxia remains one of the main causes of later disability in term infants. Despite many publications identifying possible predictors of outcome in this population of interest, little is known of the long-term developmental outcome of asphyxiated term neonates. Observational studies have largely focused on short term outcomes, with an emphasis on significant neurologic sequelae and intellectual impairments. This article reviews the literature that has described the developmental outcome of asphyxiated term newborns. As part of this review, we have also highlighted the evolution of the definition of asphyxia and delineated appropriate markers that should be used in future research on this population. PMID- 11732763 TI - Genetic factors in athetoid cerebral palsy. AB - Within the cerebral palsy syndromes, athetosis is most commonly causally associated with serious perinatal complications. Genetic factors are thought to play a lesser role, although the risk of recurrence in siblings has been suggested to be as high as 10%. We have conducted a clinical study of 22 subjects with a diagnosis of athetoid cerebral palsy and a review of the literature aiming to identify instances of familial recurrence of athetoid cerebral palsy. The birth history, family history, and previous investigations of subjects with athetoid cerebral palsy were studied and subjects were clinically examined for evidence of an underlying genetic etiology. Factors suggesting a genetic cause were specifically sought, such as advanced paternal age, progression of symptoms, and associated congenital abnormalities. No subjects in the study group had similarly affected relatives, and additional features suggesting a genetic cause were not observed. A literature search identified 16 instances of familial recurrence of athetoid cerebral palsy. Familial cases were typically associated with significant spasticity, microcephaly, intellectual disability, seizures, and a lack of history of birth asphyxia, and most could be explained by either autosomal-recessive or X-linked-recessive inheritance. The genetic contribution to athetoid cerebral palsy is small, with an overall risk of recurrence in siblings of about 1%. This risk is lower than previously suggested in the literature. PMID- 11732764 TI - Patterns of cerebral glucose metabolism in early and late stages of Rasmussen's syndrome. AB - Rasmussen's syndrome is a chronic encephalitis characterized by intractable focal epilepsy and progressive neurologic deterioration with lateralized brain destruction. In the early stages of the disease, the diagnosis can be difficult to make, and brain biopsy is often performed. We evaluated the patterns of cerebral glucose metabolism using 2-deoxy-2-[18F]-fluoro-D-glucose positron emission tomography (PET) in 15 children (age range 2.9-15.4 years, mean age 8.7 +/- 4.3 years) with Rasmussen's syndrome. In 6 patients evaluated early (< or = 1 year of onset of seizures), the PET scan showed areas of abnormal metabolism restricted mostly to the frontal and temporal regions, whereas the posterior cortex was preserved. Pathologic changes seen in the resected cortex were more pronounced in cortical areas of abnormal metabolism than in regions showing normal metabolism. In 9 patients evaluated later (>1 year after onset of seizures), the PET scan showed more diffuse hemispheric metabolic abnormalities including the occipital cortex, but the abnormalities remained highly lateralized. These patterns of glucose metabolic abnormalities in the early and late stages of the disease may facilitate the diagnosis of Rasmussen's syndrome and assist guidance of biopsy in early cases, when structural neuroimaging is still normal. PMID- 11732765 TI - Clinical experience with topiramate dosing and serum levels in children 12 years or under with epilepsy. AB - Only a limited topiramate dosing range (5-9 mg/kg/day) is approved by the U.S. Food and Drug Administration (FDA). We reviewed our topiramate dosing (mg/kg/d) and corresponding serum levels (microg/mL) (n = 77) in 41 children who were treated to clinical response or tolerability. The patients were divided into older (6-12 years [n = 21]) and younger (< or = 5 years [n = 20]) groups. Topiramate was given as monotherapy (n = 9), with an enzyme-inducing antiepileptic drug (n = 16) (phebarbital, phenytoin, or carbamazepine), or as polytherapy (n =17) (another antiepileptic drug). In the older children, there was a good dosage to serum level correspondence. However, younger children on topiramate monotherapy or cotherapy with an enzyme-inducing antiepileptic drug had relatively lower serum levels, but the serum level was increased if they were on polytherapy without an enzyme-inducing drug. This study supports a wider dosing range (7-22 mg/kg/day) of topiramate and dosage escalation beyond the approved range. Serum levels are useful in guiding topiramate dosing, especially in young children. PMID- 11732766 TI - Etiology and pathophysiology of autistic behavior: clues from two cases with an unusual variant of neuroaxonal dystrophy. AB - Two unrelated individuals with autistic behavior had numerous swollen axon terminals (spheroids) located in specific brain regions relevant to their behavioral symptoms. Spheroids are characteristic of neuroaxonal dystrophy, but the clinical profile and anatomic distribution of the lesions in these two patients differed from those of previously described patients with neuroaxonal dystrophy. Spheroids were numerous in the sensory nuclei of the spinal cord and medulla, specific nuclei and the reticular formation of the brainstem tegmentum, hypothalamus, anterior and dorsomedial thalamus, hippocampus, and cingulate and orbitofrontal cortices. Spheroids were sparse in the primary and association cortices and basal ganglia and absent in the hemispheric white matter. Cerebellar atrophy was present in both cases but associated with spheroids in only one case. These cases represent a new variant of neuroaxonal dystrophy in which behavioral symptoms characteristic of autism dominated the clinical picture. Neuroaxonal dystrophy should be included in the list of diseases that may be found in persons with autism. PMID- 11732767 TI - Seizure control and educational outcome in childhood-onset epilepsy. AB - Patients with epilepsy are more prone to have learning disabilities. This study investigated the therapeutic and educational outcome of 102 epileptic children. Analyzed data included age at onset, etiology, presence of underlying brain lesions, seizure type, and electroencephalographic (EEG) patterns. Cryptogenic seizures, remote symptomatic seizures, and underlying brain lesions were found in 29, 26, and 14 patients, respectively, whereas 47 patients had idiopathic seizures. Eighty-three patients achieved seizure control (46 remained seizure free), and 19 patients remained poorly controlled. Sixty-five patients were in regular schools, and 37 required special education (17 with mental retardation). Predictors for poor seizure control were remote symptomatic seizures, underlying brain lesions, and (when grouped together) hypsarrhythmia and mixed EEG patterns (P < .001). Predictors for special education needs were young age at onset, remote symptomatic seizures, underlying brain lesions, hypsarrhythmia and mixed EEG patterns, and poor seizure control (P < .001). We conclude that in childhood epilepsy, the need for special education is substantial and more common than treatment failure. PMID- 11732768 TI - Muromonab-CD3-induced neurotoxicity: report of two siblings, one of whom had subsequent cyclosporin-induced neurotoxicity. AB - Muromonab-CD3 is widely used for immunosuppression in patients undergoing solid organ transplant. We report two siblings with oligomeganephronia and end-stage renal disease who developed encephalopathy and seizures from muromonab-CD3 following renal transplant. The first case is a 13-year-old girl who developed encephalopathy, seizure, and triparesis following renal transplant while muromonab-CD3 was used for immunosuppression. The second case was the 6-year-old sister of the first case, who also developed recurrent focal seizures while she was on muromonab-CD3 for renal transplant immunosuppression. In both cases, a sequential brain magnetic resonance image (MRI) showed progression of abnormalities from the cerebral cortex to the white matter. In the first case, the MRI normalized after muromonab-CD3 was discontinued. In the second case, the patient developed a leukoencephalopathy following cyclosporin administration. The pathophysiology of muromonab-CD3 encephalopathy is believed to be a disturbance to the blood-brain barrier mediated by cytokine release from lymphocyte stimulation by muromonab-CD3. Because the major histocompatibility complex genes are known to regulate cytokine responses, it is possible that the excessive production of cytokines that causes encephalopathy may occur in patients who share close major histocompatibility complex genes. Muromonab-CD3 in a patient whose sibling has developed cerebral complications from its use should be administered with caution. The second case suggests that muromonab-CD3 encephalopathy predisposes patients to develop cyclosporin neurotoxicity. Because the pathogenesis of muromonab-CD3 encephalopathy and cyclosporin-related cerebral complications are both potentially mediated through a disturbance of the blood brain barrier, it is possible that one agent may predispose a patient to the complication of the other. PMID- 11732769 TI - Study of a group of extremely preterm infants (25-27 weeks): how do they function at 1 year of age? AB - The functional outcome of 49 extremely preterm infants (gestational age: 25-27 weeks) was studied at the corrected age of 12 months. Apart from pediatric follow up, a full neurologic assessment and the Bayley Motor and Mental Scales of Infant Development was done. Emphasis was placed on postural control, spontaneous motility, hand function, and elicited infantile reactions. Special attention was given to symmetric development. The infants were then categorized as having optimal or nonoptimal or asymmetric outcome. Overall, an optimal outcome was found in 19 infants (39%) and nonoptimal outcome in 30 infants (61%), 7 of whom failed on all domains of function. Postural control had a significant influence on the different domains of development such as motility (P < or = .001) and persistent infantile reactions (P < or = .001) and slightly less on hand function (P = .08) and asymmetry (P = .06). The outcome on spontaneous motility was significantly related to the results on infantile reactions (P < or = 005) and hand function (P = .05). Also, the score on the motor scale of the Bayley Developmental test was clearly related to outcome on spontaneous motility (P < or = .001) and reactions (P< or = .02). Abnormal brain ultrasonograms were related to the asymmetry of the infantile reactions (P < or = .05). Poor coordination of gross motor function will have consequences for appropriate visuomotor and sensorimotor integration, thereby hampering motor learning and later cognitive function, as is often described in preterm infants. It is suggested that the poor postural control found in many infants born preterm is the result of both myogenic and neurogenic deviations caused by the preterm birth and its nursing consequences. PMID- 11732770 TI - Prenatal intracranial hemorrhage and neurologic complications in alloimmune thrombocytopenia. AB - Neonatal alloimmune thrombocytopenia results from platelet-antigen incompatibility between mother and fetus, leading to antibody-mediated destruction of fetal platelets. With a prevalence of 1 in 1000 births, approximately 4000 infants born in the United States each year develop neonatal alloimmune thrombocytopenia. Ten to 20% of affected neonates develop intracranial hemorrhage, with 25 to 50% occurring prenatally. We report three infants who developed prenatal hemorrhage. One died in utero, and the other two had cerebral porencephaly and neurologic deficits. Infants with neonatal alloimmune thrombocytopenia have elevated risks of perinatal death and neurologic complications, including cerebral palsy, hypotonia, cortical blindness, developmental delay, seizures, and psychomotor retardation. We also report our retrospective review of the New England Medical Center neonatal intensive care unit between 1990 and 1999. Using current management guidelines, including treatment of the mother with a weekly infusion of high-dose (1-2 g/kg) intravenous immunoglobulin and/or corticosteroids, all eight infants with neonatal alloimmune thrombocytopenia did well. PMID- 11732771 TI - Vagus nerve stimulation therapy in pediatric patients with refractory epilepsy: retrospective study. AB - This six-center, retrospective study evaluated the effectiveness, tolerability, and safety of vagus nerve stimulation in children. Data were available for 125 patients at baseline, 95 patients at 3 months, 56 patients at 6 months, and 12 patients at 12 months. The typical patient, aged 12 years, had onset of seizures at age 2 years and had tried nine anticonvulsants before implantation. Collected data included preimplant history, seizures, implant, device settings, quality of life, and adverse events. Average seizure reduction was 36.1% at 3 months and 44.7% at 6 months. Common adverse events included voice alteration and coughing during stimulation. Rare adverse events, unique to this age group, included increased drooling and increased hyperactivity. Quality of life improved in alertness, verbal communication, school performance, clustering of seizures, and postictal periods. We concluded that vagus nerve stimulation is an effective treatment for medically refractory epilepsy in children. PMID- 11732772 TI - NeuroD homologue expression during cortical development in the human brain. AB - Neurogenesis and neuronal differentiation are determined by the NeuroD homologues, transcription factors belonging to a family of basic helix-loop-helix proteins. The authors used in situ hybridization with full-length riboprobes for NeuroD1, NeuroD2, and NeuroD3 to describe the expression of the NeuroD homologues in a gestational sequence of human fetal brains. Acridine orange histofluorescence was used to differentiate neuronal from non-neuronal cell precursors. At the earliest gestational age examined (gestational week 16), signals for all three homologues could be identified but that for NeuroD3 was most intense. Peak expression of NeuroD1 and NeuroD2 followed at gestational weeks 19 and 20, respectively. Although similar to the expression of these homologues in the mouse cerebrum, notable differences were observed. Specifically, signals for all three homologues were detected in the marginal zone and the ventricular zone, including the ganglionic eminence. The temporal order of expression in the human is similar to that in the mouse, in spite of these anatomic differences. These data are consistent with NeuroD3 serving as a determination factor, which commits the post-mitotic progenitor cell to a neuronal fate, whereas NeuroD1 and NeuroD2 appear more likely to play a role in neuronal differentiation. PMID- 11732773 TI - Psychometric testing in bacterial meningitis: results of a long-term prospective study of infants and children treated between 1973 and 1977. PMID- 11732774 TI - Congenital hydranencephalic-hydrocephalic syndrome with proliferative vasculopathy: a possible relation with mitochondrial dysfunction. AB - We report the case of a fetus aborted at gestation week 20 because of hydranencephalic-hydrocephalic syndrome. The fetus was the third pregnancy of a nonconsanguineous couple whose first child exhibited congenital hydranencephalic hydrocephalic syndrome associated with muscle histology findings consistent with mitochondrial cytopathy and deficiency of complexes III and IV of the respiratory chain and whose second pregnancy had terminated in an elective abortion on detection of progressive hydrocephalus at gestation week 19. The third pregnancy had a normal course according to obstetric and ultrasonography examinations performed at gestation weeks 5, 10, and 15, and negative results were obtained in standard serologic and polymerase chain reaction (PCR) tests for prenatal infections of the mother. However, the ultrasonography examination at gestation week 18 revealed hydrocephalus, in response to which the parents requested an abortion, which was performed at gestation week 20; the fetus was male and with no evident external malformations. Histopathologic studies of the brain and medulla oblongata revealed proliferative vasculopathy (glomeruloid vessels, intracytoplasmic inclusions, and microcalcifications) and intracytoplasmic inclusions in the voluntary muscle. Microbiologic and PCR tests of hepatic and spleen tissue were negative for prenatal infections. In view of the precedent of a sister with mitochondrial dysfunction, these findings raise the pos sibility that at least some cases of familial syndrome of congenital hydranencephalic hydrocephalic syndrome with proliferative vasculopathy can be attributed to alterations in the mitochondrial respiratory chain. PMID- 11732776 TI - Enteroviral encephalitis leading to a locked-in state. AB - Millions of children are infected by enteroviruses each year, usually exhibiting only mild symptoms. Although enteroviruses are a common cause of community acquired aseptic meningitis, enteroviral meningitis usually has a benign course. We describe a 14-year-old patient with enteroviral meningoencephalitis diagnosed by polymerase chain reaction. Her level of consciousness declined rapidly after hospital admission and resulted in a locked-in state. Although she experienced slow neurologic improvement, residual neurologic deficits remain. Although there is a general awareness of the characteristics of enteroviralencephalitis, this case report is significant in presenting a case of unusual severity and sequelae. To our knowledge, this is the first published report of an enterovirus encephalitis leading to a locked-in-state. PMID- 11732775 TI - Activated protein C resistance and false type 2 protein C deficiency detected after multiple shunt failures in a patient with hydrocephalus. AB - A 2-year-old hydrocephalic boy who had suffered multiple shunt failures was evaluated for hypercoagulability after a thrombus was removed from his right atrium. The work-up revealed that the patient had the heterozygous form of activated protein C resistance and false type 2 protein C deficiency by the clotting method. His protein C activity was normal by the chromogenic method. We suggest that patients having hydrocephalus, shunt-associated thrombus formation, or both should be evaluated for thrombophilic disorders, and protein C activity should be measured by chromogenic assay in patients with documented activated protein C resistance. PMID- 11732777 TI - Transverse myelitis associated with herpes simplex virus infection. AB - The case of an otherwise healthy 6-year-old boy with symptoms and signs of acute transverse myelitis is presented. The diagnosis was confirmed by magnetic resonance imaging. An episode of gingivostomatitis had preceded, and serology indicated herpes simplex virus type 1 infection. The child recovered gradually, and no relapses were noted during a 30-month follow-up period. PMID- 11732778 TI - Cerebral mycotic aneurysm in a child with Down's syndrome: a unique association. AB - Mycotic aneurysms are rare complications in patients with infective endocarditis, particularly in the pediatric population. We report a case of mycotic aneurysm of the middle cerebral artery complicating bacterial endocarditis in a child with Down's syndrome. The patient was successfully treated medically without the need for surgical intervention. PMID- 11732779 TI - Prednisone therapy in Becker's muscular dystrophy. AB - Two boys with Becker's muscular dystrophy had a dramatic and sustained improvement in strength with therapeutic use of pred nisone. Both had documented Xp-21 defects on DNA testing. Concurrently with improvement, there was a decrease in their serum creatine kinase levels. One patient had two muscle biopsies, the first before prednisone treatment and the second during treatment. Both biopsies demonstrated an immunoblot decrease in the quantity of muscle dystrophin. Routine histology on the first biopsy was consistent with muscle dystrophy, and the second biopsy was normal. These two patients suggest that a small percentage of patients with Becker's muscular dystrophy have a dramatic and sus tained improvement in strength with the use of therapeutic corticosteroids. All patients with Becker's muscular dystrophy should be given a careful trial of prednisone to define those who benefit. PMID- 11732780 TI - Genotype analysis of methicillin-resistant Staphylococcus aureus with and without reduced susceptibility to vancomycin using pulsed-field gel electrophoresis. PMID- 11732781 TI - Requiem for reuse of single-use devices in US hospitals. PMID- 11732782 TI - Decontaminated single-use devices: an oxymoron that may be placing patients at risk for cross-contamination. AB - OBJECTIVE: To determine whether reprocessed single-use devices would meet regulatory standards for sterility and meet the same materials standards as a new device. DESIGN: The study included single-use and reusable biopsy forceps and papillotomes and a reusable stone retrieval basket. The suitability of these devices for cleaning and disinfection or sterilization was examined. METHODS: Testing of cleanability was conducted on devices contaminated with technetium 99 radiolabeled human blood. Instruments were cleaned using hospital recommended practices for manual cleaning. Gamma counts per second were determined before and after cleaning to localize contaminants, which were additionally visualized using light and scanning electron microscopy. X-ray photoelectron spectroscopy was used to quantify contamination elements on the materials tested. Residual bioburden testing on instruments contaminated with microorganisms suspended in coagulable sheep blood was carried out to establish the efficacy of disinfection and sterilization. RESULTS: All devices remained contaminated after cleaning, but single-use devices and the stone basket tended to be more heavily contaminated than reusable forceps and papillotomes. Cleaning procedures facilitated distribution of contaminants further into the lumens of the disposable forceps. Decreased concentrations of silicon and increased concentrations of carbon and nitrogen suggested that layers of silicon lubricant had been removed and contaminants were organic material. Reusable devices were effectively disinfected, but single-use devices were not. Sterilization could not eliminate the challenge microorganisms completely. CONCLUSIONS: None of the reprocessed single-use instruments were effectively cleaned, disinfected, or sterilized. This condition may provide an opportunity for the viability of nonresistant or nosocomial organisms and viruses. Additionally, reprocessing procedures may result in material destruction of fragile devices. Cost-saving initiatives that have inspired reprocessing of single-use devices, despite the absence of data establishing the efficacy of decontamination and the durability of materials throughout reprocessing, should be pushed into the background. PMID- 11732783 TI - Comparison of pulsed-field gel electrophoresis and amplified fragment-length polymorphism for epidemiological investigations of common nosocomial pathogens. AB - OBJECTIVE: To compare molecular typing by amplified fragment-length polymorphism (AFLP) analysis with pulsed-field gel electrophoresis (PFGE) with respect to the ability to differentiate between epidemiologically related and unrelated isolates of common nosocomial pathogens recovered during a period of endemicity. DESIGN: Retrospective laboratory analysis. SETTING: Tertiary-care institution. METHODS: 17 isolates of Acinetobacter baumannii, 22 isolates of Pseudomonas aeruginosa, and 22 vancomycin-resistant Enterococcusfaecium (VRE) were typed by both methods. RESULTS: AFLP generated comparable results to PFGE for A baumannii and P aeruginosa isolates; both methods identified epidemiologically related and unrelated isolates. However, strain typing of VRE isolates produced discordant results between the two methods. PFGE identified 10 different strain types and differentiated between all epidemiologically related and unrelated isolates. In contrast, AFLP generated only five different strain types, three of which contained both epidemiologically related and unrelated isolates. CONCLUSION: Molecular typing by AFLP is comparable to PFGE for A baumannii and P aeruginosa isolates. For VRE isolates, however, PFGE remains the method of choice. PMID- 11732784 TI - Infection control practices among correctional healthcare workers: effect of management attitudes and availabiity of protective equipment and engineering controls. AB - OBJECTIVES: To determine the relation of the availability of personal protective equipment (PPE) and engineering controls to infection control (IC) practices in a prison healthcare setting, and to explore the effect on IC practices of a perceived organizational commitment to safety. DESIGN: Cross-sectional survey. SETTING: The study population was drawn from the 28 regional Correctional Health Care Workers Facilities in Maryland. PARTICIPANTS: All full-time Maryland correctional healthcare workers (HCWs) were surveyed, and 225 (64%) of the 350 responded. METHOD: A confidential, self-administered questionnaire was mailed to all correctional HCWs employed in the 28 Maryland Correctional Health Care Facilities. The questionnaire was analyzed psychometrically and validated through extensive pilot testing. It included items on three major constructs: IC practices, safety climate (defined as the perception of organizational commitment to safety), and availability of IC equipment and supplies. RESULTS: A strong correlation was found between the availability of PPE and IC practices. Similarly, a strong correlation was found between IC practices and the presence of engineering controls. In addition, an equally strong association was seen between the adoption of IC practices and employee perception of management commitment to safety. Those employees who perceived a high level of management support for safety were more than twice as likely to adhere to recommended IC practices. IC practices were significantly more likely to be followed if PPE was always readily available. Similarly, IC practices were more likely to be followed if engineering controls were provided. CONCLUSION: These findings suggest that ready availability of PPE and the presence of engineering controls are crucial to help ensure their use in this high-risk environment. This is especially important because correctional HCWs are potentially at risk of exposure to bloodborne pathogens such as human immunodeficiency virus and hepatitis B and C viruses. Commitment to safety was found to be highly associated with the adoption of safe work practices. There is an inherent conflict of "custody versus care" in this setting; hence, it is especially important that we understand and appreciate the relation between safety climate and IC practices. Interventions designed to improve safety climate, as well as availability of necessary IC supplies and equipment, will most likely prove effective in improving employee compliance with IC practices in this healthcare setting. PMID- 11732785 TI - Contamination of gowns, gloves, and stethoscopes with vancomycin-resistant enterococci. AB - OBJECTIVE: [corrected] To measure directly the rate of contamination, during routine patient examination, of gowns, gloves, and stethoscopes with vancomycin resistant enterococci (VRE). SETTING: A large, academic, tertiary-care hospital. PATIENTS: Between January 1997 and December 1998, 49 patients colonized or infected with VRE were entered in the study. DESIGN: After routine examination, the examiner's glove fingertips, gown (the umbilical region and the cuffs), and stethoscope diaphragm were pressed onto Columbia colistin-nalidixic acid (CNA) agar plates with 5% sheep blood plus vancomycin 6 pg/mL. The stethoscope diaphragm was sampled again after cleaning with a 70% isopropanol wipe. RESULTS: VRE were isolated from at least 1 examiner site (gloves, gowns, or stethoscope) in 33 (67%) of 49 cases. Gloves were contaminated in 63%, gowns in 37%, and stethoscopes in 31%. All three items were positive for VRE in 24%. One case each had stethoscope and gown contamination without glove contamination. Only 1 (2%) of 49 stethoscopes was positive after wiping with an alcohol swab. Contamination at any site was more likely when the patient had a colostomy or ileostomy. Patients identified by rectal-swab culture alone were as likely to contaminate their examiners as were those identified by clinical specimens. CONCLUSIONS: Our study revealed a high rate of examiner contamination with VRE. The similar risk of contamination identified by surveillance and clinical cases reinforces concerns that patients not known to be colonized with VRE could serve as sources for dissemination. Wiping with alcohol is effective in decontaminating stethoscopes. PMID- 11732786 TI - Correlates of antibiotic use in Taiwan hospitals. AB - OBJECTIVE: To determine factors that correlate with increased antibiotic use among adult inpatients in Taiwan. DESIGN: Retrospective survey of medical records. SETTING: 14 acute-care hospitals (8 regional hospitals, 6 medical centers) in Taiwan. PARTICIPANTS: A systematic probability sample from each hospital, totaling 663 adult inpatients who were discharged or had died in early 1999. MEASUREMENTS: Infectious disease physicians at the 14 hospitals collected data from medical records regarding patient demographics, hospitalization, discharge diagnosis, and antibiotics received. RESULTS: A total of 447 (67%) patients received antibiotics for an overall rate of 813 antibiotic-days (number of days patients received each antibiotic)/1,000 patient-days. Both the proportion of beds in intensive care units ([ICUs] Pearson correlation coefficient [r], 0.67; 95% confidence interval [CI 95], 0.36-0.89; P<.01) and the proportion of patients admitted to surgical services (r, 0.66; CI 95, 0.20-0.88; P=.01) correlated with the mean patient rate of antibiotic-days/hospital-day (MPAUD). In contrast, we found no correlation between the proportion of patients who received antibiotics and the MPAUD. Using multiple linear regression, medical center status was the only independent predictor for increased MPAUD (regression coefficient [b], 0.15; CI 95, 0.05-0.24; P<.01). There was no correlation between pooled rates of antibiotic-days/hospital-day and any hospital demographic factors. First-generation cephalosporin (39%) and aminoglycoside (24%) use accounted for the majority of antibiotic-days. CONCLUSIONS: Antibiotic use is greater in medical centers than in regional hospitals and appears to be independent of surgical case mix or the proportion of ICU beds. Determination of multiple, independent measures of antibiotic use may be necessary to understand the relation between antibiotic use and resistance in hospital. PMID- 11732787 TI - Quinolone use as a risk factor for nosocomial Clostridium difficile-associated diarrhea. AB - OBJECTIVE: To determine modifiable risk factors for nosocomial Clostridium difficile-associated diarrhea (CDAD). DESIGN: Case-control study. SETTING: 300 bed tertiary-care hospital. PARTICIPANTS: Hospital inpatients present during the 3-month study period. METHODS: Case-patients identified with nosocomial CDAD over the study period were compared to two sets of control patients: inpatients matched by age, gender, and date of admission; and inpatients matched by duration of hospital stay. Variables including demographic data, comorbid illnesses, antibiotic exposure, and use of gastrointestinal medications were assessed for case- and control-patients. Conditional logistic regression was performed to identify risk factors for nosocomial CDAD. RESULTS: 27 case-patients were identified and were compared to the two sets of controls (1:1 match for each comparison set). For the first set of controls, use of ciprofloxacin (odds ratio [OR], 5.5; 95% confidence interval [CI 95], 1.2-24.8; P=.03) was the only variable that remained significant in the multivariable model. For the second set of controls, prior exposure to cephalosporins (OR, 6.7; CI 95, 1.3-33.7; P=.02) and to ciprofloxacin (OR, 9.5; CI 95, 1.01-88.4; P=.05) were kept in the final model. CONCLUSIONS: Along with cephalosporins, prior quinolone use predisposed hospitalized patients to nosocomial CDAD. Quinolones should be used judiciously in acute-care hospitals, particularly in those where CDAD is endemic. PMID- 11732788 TI - The prevalence of fecal colonization with VRE among residents of long-term-care facilities in Melbourne, Australia. AB - A point-prevalence survey performed among residents of eight nursing homes in Melbourne, Australia, found a rate of fecal VRE colonization of 3.1% (9/292; 95% confidence interval, 1.1-5.1), all vanB Enterococcusfaecium. This is a higher rate than in the general community (3.1% vs 0.2%). Many residents (16%) had been inpatients in acute-care hospitals in the previous 3 months. PMID- 11732789 TI - Topical mupirocin for eradication of MRSA colonization with mupirocin-resistant strains. AB - Topical mupirocin was able to interrupt colonization of 52% and 68% of methicillin-resistant Staphylococcus aureus (MRSA)-colonized patients carrying mupirocin-resistant and -sensitive strains, respectively, including 44.4% and 85.7% of those colonized only in the nares. Although a trend to decreased effectiveness was seen for clearing mupirocin-resistant MRSA, this agent can decolonize many patients with resistant strains. PMID- 11732790 TI - A shared vision of healthcare quality improvement. AB - SHEA and the American Society for Quality's Health Care Division have been collaborating in areas of common concern to improve healthcare quality. We each possess a heritage of different but complementary approaches and stand a better chance of success together than apart. This presentation describes rapid growth of our interdisciplinary, international, special interest group and progress made thus far, as well as challenges facing hospital epidemiologists and quality improvement professionals. PMID- 11732791 TI - Models and mechanisms for building and funding partnerships. AB - The Canadian Institutes of Health Research (CIHR) recently replaced two former federal agencies, the Medical Research Council and the National Health Research and Development Program. CIHR has attracted widespread international attention as a grand experimental model that emphasizes excellence, interdisciplinarity, networking, partnership, and a collective process for priority setting, alongside continuing support for the best ideas of the most talented investigators. CIHR identified four cross-cutting health research themes (biomedical research; clinical research; health service; systems research; and research into the societal, cultural, and environmental determinants of health) across its 13 virtual institutes. Each institute is required to bring together investigators working in specific aspects of health research, as they relate to the mandate of the institute, in order to stimulate novel interdisciplinary research approaches. The challenge to Canadian members of the Society for Healthcare Epidemiology of America and their collaborators from other countries is to engage in partnership with CIHR, so that together we can ensure unparalleled quality, safety, and efficacy of healthcare systems in Canada and around the world. PMID- 11732792 TI - Experience in the field: assessing quality of health care reform in Canada. AB - The Canadian Institute for Health Information (CIHI) maintains databases to report independently on the health of Canadians and the performance of their healthcare system. CIHI recently received funding to improve indicator development for tracking quality, because in Canada we really have very few data that allow us to make comparable observations down to a regional or institution specific interest level. We have a long way to go, and we recognize that collaboration and partnership is the only way that this development will move ahead. PMID- 11732793 TI - CD1--the pathology perspective. AB - CD1 molecules are a family of cell surface-associated glycoproteins now recognized as having a role in antigen presentation. These glycoproteins are distinct from yet have some similarities to classical major histocompatibility complex class I and class II molecules. The role of these molecules has been studied in detail over recent years, with an explosion of interest following the demonstration that they can present nonprotein antigens to certain subpopulations of T cells. The purpose of this paper is to provide an overview of current knowledge of the function of the CD1 family with specific emphasis on the potential role in the pathogenesis of certain diseases. Although much of the current research in this field has inevitably concentrated on mice and humans, this work also has potential significance for veterinary species. PMID- 11732794 TI - Granular cell tumor of the canine central nervous system: two cases. AB - Although pure granular cell tumors have been reported in various sites in the dog, only one tumor has been reported in the central nervous system. Two dogs presented with neurologic signs had brain lesions detected by magnetic resonance imaging in the area of the olfactory bulbs and frontal cortex. In both dogs, a clinical diagnosis of a granular cell tumor was made from tissues obtained from stereotactic biopsies guided by computed tomography. Surgical removal of the tumors was followed by histopathologic, ultrastructural, and immunocytochemical characterization. Although not conclusive, these studies indicated that the granular cells were not of leucocyte origin but may have been derived from the meninges. One dog died 12 months after surgery, and the other was alive 4 months later. PMID- 11732795 TI - Characterization of the inflammatory infiltrate in canine chronic hepatitis. AB - Canine chronic hepatitis (CCH) is a progressive inflammatory disease of unknown etiology. To characterize the inflammatory infiltrate, 16 dogs with CCH were selected and classified into three groups based on the stage of fibrosis, as evaluated with Masson's trichrome stain. The inflammatory infiltrate in each liver section was immunohistochemically characterized and evaluated using CD3, lysozyme, lamba and kappa light chain, and alpha-smooth muscle actin antibodies. Numerous breeds were affected, and middle-aged females predominated in this select group. Necroinflammatory activity progressively increased and then waned as the hepatitis progressed to cirrhosis. CD3+ lymphocytes were the most numerous lymphoid cells in dogs with CCH. Degenerate hepatocytes were occasionally surrounded by CD3+ lymphocytes. Necrosis was positively correlated with the number of CD3+ lymphocytes. The lamba and kappa light chain-positive cell infiltrate was variable but generally mild. A positive correlation between the lambda and kappa light chain-positive cells and the portal alpha-smooth muscle actin was found. The number of alpha-smooth muscle actin-positive cells (myofibroblasts) in portal triads and fibrous septa was positively correlated with the stage of fibrosis. In contrast, no correlation was found between the number of lysozyme-positive cells (Kupffer cells) and the stage of fibrosis. These results further support the idea of an immune-mediated process in CCH and suggest that periductular myofibroblasts play an important role in canine liver fibrogenesis. PMID- 11732796 TI - Immunohistochemical characterization of canine hyperplastic hepatic lesions and hepatocellular and biliary neoplasms with monoclonal antibody hepatocyte paraffin 1 and a monoclonal antibody to cytokeratin 7. AB - Immunostaining with monoclonal antibody (MoAb) hepatocyte paraffin 1 (Hep Par 1) and an MoAb to cytokeratin 7 (CK7) was performed on 105 formalin-fixed, paraffin embedded canine hyperplastic and neoplastic hepatic lesions. Hep Par 1 was detected in 12/12 hyperplastic nodules, 17/17 hepatocellular adenomas, and 37/40 hepatocellular carcinomas. The staining was disseminated, granular, and cytoplasmic. This antibody did not react with normal or neoplastic biliary epithelium. Other hepatic tumors or tumors metastatic to the liver did not bind Hep Par 1 except one metastatic intestinal carcinoma. MoAb to CK 7 stained all hyperplastic biliary epithelium and benign cholangiocellular tumors (5/5) and 14/18 cholangiocellular carcinomas. One hepatocellular carcinoma had cells positive for both Hep Par 1 and CK 7. Liver was the only normal tissue tested that reacted with MoAb Hep Par 1. Only five nonhepatic tumors (one adrenocortical carcinoma, one interstitial cell tumor of the testis, one melanoma, and two salivary adenocarcinomas) of 277 tumors tested had focal/multifocal staining for Hep Par 1. Prolonged fixation did not alter the staining with Hep Par 1. We conclude that Hep Par 1 is a specific and sensitive marker for canine hepatocellular tumors and allows distinction between hepatocellular and biliary neoplasms. PMID- 11732797 TI - Ovine adenovirus serotype 7-associated mortality in lambs in the United States. AB - Adenoviral infections were diagnosed in three neonatal lambs that died spontaneously, and no other etiologic agents were identified. Clinical signs were anorexia, weakness, abdominal distention, and sudden death. Microscopic lesions consisted of multifocal necrotizing hepatitis, multifocal subacute interstitial nephritis, and loss of enterocytes from intestinal villi. Adenovirus inclusions were identified by light microscopy in the kidneys only. Adenoviral antigen, however, was identified in the liver, kidney, and intestine of the lambs by immunohistochemical techniques. An ovine adenovirus serotype 7, not previously isolated from sheep in the United States, was characterized from these lambs. PMID- 11732798 TI - Localization of avian leukosis virus subgroup J in naturally infected chickens by RNA in situ hybridization. AB - The novel subgroup J of avian leukosis virus (ALV-J) has emerged as a significant cause of myeloid neoplasia and weight suppression in broiler chickens. We investigated viral tropism using RNA in situ hybridization (ISH) in naturally infected chickens. Formalin-fixed tissues were collected from 12-day-old embryos (seven infected, two control) and from 0-week-old (four infected, one control), 3 week-old (five infected, one control), 6-week-old (five infected, one control), and 9-week-old (10 infected, two control) chickens naturally infected with ALV-J in ovo. A 636-base antisense riboprobe complementary to the 3' and 5' ends of the pol and env viral genes, respectively, was constructed. Strong positive staining was present in cardiac myocytes, Purkinje fibers, vascular and pulmonary smooth muscle, renal glomeruli, distal tubules, and pituitary glands. Light staining was present in gastrointestinal smooth muscle, thyroid and adrenal glands, and follicular medullae in the cloacal bursa. Staining was not present in any hematopoietic precursors. Tissues from newly hatched chicks exhibited the strongest and most consistent staining, whereas staining in embryos was minimal. RNA ISH confirmed the presence of ALV-J-specific nucleic acid within cytoplasmic inclusions in cardiac myocytes, Purkinje fibers, pituitary glands, and renal glomeruli. Viral tropism for cardiac myocytes and Purkinje fibers may relate pathogenetically to the cardiomyopathy and congestive heart failure described in index chicken flocks infected with ALV-J. Viral tropism for endocrine organs may relate pathogenetically to the weight suppression associated with infection. PMID- 11732799 TI - Uroplakin expression in the urothelial tumors of cows. AB - Expression of uroplakins (UPs) was investigated in 20 bladder tumors from cows that had been suffering from chronic enzootic hematuria for several years. In dysplastic urothelium and papillomatous proliferations, UP expression was evident both as luminal and intercellular staining. UPs appeared to clearly define the plasma membrane of luminal cells and the borders of cells placed in deeper layers, whether or not these intermediate cells were adjacent to superficial ones. Occasionally, some intermediate cells showed a remarkable cytoplasmic immunoreactivity. The pattern of UPs in grade I tumors was characterized by an evident discontinuity of luminal staining and by the presence of numerous intermediate cells showing a diffuse intracytoplasmic positivity for UPs. In grade II tumors, there was a decrease of luminal and intermediate cells showing UP expression and an apparent increase of clusters of intermediate cells with intracytoplasmic reactivity for UPs. In grade III tumors, immunoreactivity was heterogeneously distributed and a severe loss of UP-positive luminal and intermediate cells could be seen. Focally, superficial and deeper cells showed strong membraneous immunoreactivity that marked and delimited single cells, with complete circumferential peripheral staining clearly evident. UP expression in bladder tumors of cows reported in this study is similar to the UP pattern of some urothelial tumors in humans. Although UP expression is remarkably changed in bladder carcinogenesis of cattle, the UP gene(s) remains expressed during cell transformation and tumor progression. PMID- 11732800 TI - Immunohistochemical detection of inhibin-alpha, -betaB, and -betaA chains and 3beta-hydroxysteroid dehydrogenase in canine testicular tumors and normal testes. AB - Immunohistochemical detection of inhibin-alpha, -betaA and -betaB chains and 3beta-hydroxysteroid dehydrogenase (HSD) was carried out on primary testicular tumors from 15 dogs and normal testes from three adult dogs. Histopathologically, the tumors were composed of three types: Leydig cell tumors in five dogs, Sertoli cell tumors in five dogs, and seminoma in five dogs. In normal testes, immunostaining against inhibin-alpha, -betaA, and -betaB chains and 3beta-HSD revealed positive reactivity in the cytoplasm of Leydig cells. In testicular tumors, immunoreactive cells against inhibin-alpha, -betaA, and -betaB chains and 3beta-HSD were localized in all Leydig cell tumors but not in any Sertoli cell tumors or seminomas. The results of radioimmunoassay for plasma inhibin in dogs with Leydig cell tumors showed higher concentrations than those in dogs with Sertoli cell tumors and seminomas and those in normal dogs. The concentration of inhibin in the plasma was markedly decreased by the surgical removal of the Leydig cell tumor in one dog. Our findings suggest that inhibin is synthesized by normal and neoplastic Leydig cells in the canine testis, and the secreted inhibin may be inhibin A and inhibin B. PMID- 11732801 TI - Helicobacter pylori-induced gastritis in experimentally infected conventional piglets. AB - A conventional nonmutant animal that could be experimentally infected with Helicobacter pylori isolates would be a useful animal model for human H. pylori associated gastritis. Gnotobiotic and barrier-born pigs are susceptible to H. pylori infection, but attempts to infect conventional pigs with this bacterium have been unsuccessful. In the present study, a litter of eight 20-day-old crossbreed piglets were purchased from a commercial farm. Six of them were orally challenged two to five times at different ages, between 29 and 49 days, with doses of H. pylori inoculum containing approximately 10(9) bacterial cells. Two animals served as controls. The inoculation program began 2 days postweaning when the piglets were 29 days of age. Prior to every inoculation, the piglets were fasted and pretreated with cimetidine, and prior to the first and second inoculation each piglet also was pretreated with dexamethasone. The challenged piglets were euthanasized between 36 and 76 days of age. H. pylori colonized all six inoculated piglets. The pathology of the experimentally induced gastritis was examined macroscopically and by light and electron microscopy. H. pylori induced a severe lymphocytic gastritis in the conventional piglets and reproduced the large majority of the pathologic features of the human disease. Therefore, the conventional piglet represents a promising new model for study of the various pathogenic mechanisms involved in the development of lesions of the human H. pylori-associated gastritis. PMID- 11732802 TI - Point mutation of neu oncogene in animal peripheral nerve sheath tumors. AB - Thirty-four peripheral nerve sheath tumors of four domesticated animal species were characterized and assayed for point mutation of the neu oncogene. Based on their morphoimmunophenotype, 32 tumors were classified as schwannomas. Schwannoma morphology was characterized by the presence of Antoni type A and B pattern and immunoreactivity for S-100 protein and vimentin. Two anaplastic and metastatic tumors originating from spinal cord root, immunonegative for S-100 protein and positive for vimentin, were classified as malignant peripheral nerve sheath tumors (MPNSTs). Four malignant schwannomas and two MPNSTs expressed a point mutation of the neu oncogene by the polymerase chain reaction-restriction fragment length polymorphism method. The finding of neu oncogene mutation could be a useful diagnostic genetic marker in the malignant form of peripheral nerve sheath tumors in animals. PMID- 11732803 TI - Gastrointestinal stromal tumors in equids. AB - Eleven gastrointestinal neoplasms from 10 aged horses and 1 pony were examined grossly, his tologically, immunohistochemically, and (in two cases) ultrastructurally. Clinical signs were associated with two neoplasms, and the other nine tumors were incidental findings at laparotomy or necropsy. The neoplasms were solitary (9/11) or multifocal (2/11), well demarcated, serosal or mural masses of stomach (1), jejunum (1), ileum (3), cecum (5), and/or colon (2). Microscopic examination revealed discrete spindle cells arranged in compact patterns with fascicles and whorls or cribriform pattern with fascicles and rare palisades, often with a myxoid interstitial matrix. Three tumors infiltrated between the muscularis interna and the muscularis externa at the myenteric plexi. All neoplasms were vimentin positive, 3/11 were S-100 positive, 2/11 were muscle actin positive, and no neoplasm was positive for glial fibrillary acid protein, desmin, factor VIII, chromogranin, or neuron-specific enolase. Of the two tumors studied ultrastructurally, one contained an admixture of smooth muscle cells and cells resembling Schwann cells, and the second was populated by homogeneous fusiform mesenchymal cells separated by homogeneous matrix. Gastrointestinal stromal tumors (GIST) have been recognized in humans, more recently in dogs and nonhuman primates, and now in equids. Most of these tumors are comprised of a loosely arranged network of spindled cells separated by myxoid matrix. GIST may be composed of myogenic, neurogenic, combined myogenic and neurogenic, and undifferentiated mesenchymal cells. PMID- 11732804 TI - Reduced expression of cyclin-dependent kinase inhibitor p27Kip1 in feline lymphoma. AB - Expression of p27Kip1 was identified in feline lymphoid tissues by immunohistochemistry. In normal lymphoid tissues, p27Kip1 was detected as a distinct nuclear stain in lymphocytes of the follicular mantle zone and interfollicular small lymphocytes, whereas activated lymphoblasts in the germinal center were negative. Lymphoid hyperplasia was similarly immunolabeled but with an expanded mantle zone and marginal zone of p27Kip1-reactive lymphocytes. Both T and B-cell lymphomas lacked p27Kip1 immunolabel and were determined to be proliferative based on immunohistochemical detection of the Ki-67 antigen. Scattered p27Kip1-immunolabeled lymphocytes were detected throughout the lamina propria of most specimens characterized as lymphoplasmacytic enteritis. The results of this study suggest that the antiproliferative effect of the cell cycle regulator p27Kip1 is abrogated in feline lymphoma, presumably allowing cells to bypass the G1-S checkpoint of the cell cycle. PMID- 11732805 TI - Expression of bone morphogenetic protein-6 (BMP-6) in myoepithelial cells in canine mammary gland tumors. AB - Seventy-three mammary tumors and three mammary tissue specimens were examined to elucidate the expression of bone morphogenetic protein (BMP)-6 in the myoepithelial cells of canine mammary gland tumors. Morphologically, the myoepithelial cells were classified into four types: resting and proliferating cells inside the basement membrane, and spindle- and star-shaped cells proliferating in the outer area of the basement membrane. The characteristics of these myoepithelial cells were confirmed by immunohistochemistry using antibodies raised against keratin, cytokeratin 19, alpha-smooth muscle actin, and vimentin. In simple adenoma, a small number of resting myoepithelial cells was immunopositive for BMP-6. In complex adenomas and benign mixed tumors, all types of myoepithelial cells, depending in some cases on their specific location within the tumor, were immunopositive for BMP-6, but almost all of the tubular epithelial cells were immunonegative. Foci consisting of a proliferation of BMP-6 positive star- and spindle-shaped cells had mucinous stroma with marked hyaline and chondroid changes. In contrast, the foci with BMP-6-negative spindle- and star-shaped cells tended to have mucinous stroma without chondroid change. Several types of mesenchymal cells including chondrocytes, osteoblasts, and fibroblastlike cells in the mixed tumors, showed an intense immunopositive reaction for the BMP-6 antibody, and were located close to the ectopic cartilage and bone matrix. No significant immunoreactivity for BMP-6 was observed in most of the malignant mammary tumors; only one malignant mixed tumor was examined. All of these findings indicate that BMP-6 expression in myoepithelial cells may increase in complex adenomas and benign mixed tumors in canine mammary glands, and that BMP-6 expression is most intense in the vicinity of chondroid matrix in these tumors. PMID- 11732806 TI - Piloleiomyosarcoma in seven ferrets. AB - In each of seven ferrets (Mustela putorius furo) with leiomyosarcoma, a single dermal mass was identified and biopsied. Each mass consisted of a well-demarcated but nonencapsulated proliferation of large spindle- to strap-shaped cells arranged in interwoven bundles. The cells resembled the smooth muscle cells of the adjacent arrector pili muscles, but with marked nuclear pleomorphism. Immunohistochemical staining for smooth muscle actin, desmin, and vimentin was positive and staining for myoglobin and cytokeratin was negative. Follow-up on three of the ferrets indicates that the prognosis is good following complete surgical excision. PMID- 11732807 TI - Mixed germ cell tumor in the eye of a dog. AB - A 3-year-old female neutered Staffordshire Bull Terrier presented with a mixed germ cell tumor involving the base of the iris and the ciliary body of the right eye. The tumor mass was composed primarily of packeted vacuolated, polygonal (hepatoid) cells and small round cells; epithelial cells lining tubuloacinar structures were a less prominent component. The hepatoid and round cells stained positively for alpha-fetoprotein and cytokeratin. The epithelial cells stained positively for cytokeratin only, and some contained cytoplasmic mucin droplets. The polygonal cells were interpreted as a hepatoid variant of yolk sac tumor, and the epithelial cells were considered a teratomatous component. Trabeculae of bone were observed within the mass and may have been metaplastic or a teratomatous element. Extragonadal germ cell tumors are rare in dogs and have previously been reported only in the suprasellar region. This is the first report of this tumor type in the eye of a nonhuman species. PMID- 11732808 TI - Streptozocin-induced pancreatic islet destruction in beef cows. AB - Streptozocin (STZ) induces diabetes mellitus in sheep and pigs. To test the effect of STZ in cattle, cows were given 75-150 mg STZ per kilogram of body weight. Cows receiving 150 mg/kg required euthanasia within 24 hours after infusion because of the severe systemic effects of STZ. Seven cows receiving doses of < or = 100 mg/kg had mild to marked decrease in islet immunoreactivity for insulin and in pancreatic islet density and mild to severe tubulointerstitial nephritis. Two cows receiving 75 and 85 mg/kg STZ regained their ability to produce insulin and return blood glucose to basal levels. One cow given 100 mg/kg STZ developed insulin insufficiency consistent with type I diabetes mellitus. These findings demonstrate the susceptibility of the bovine pancreas to STZ: however, severe systemic complications were encountered. Alternative dosages and methodologies should be considered in future attempts to induce diabetes in cattle using STZ. PMID- 11732809 TI - Electron microscopic observations of stratum corneum intercellular lipids in normal and atopic dogs. AB - The barrier function of mammalian skin is maintained by intercellular stratum corneum lipids. In human patients with atopic dermatitis, an abnormal lipid barrier results in dry skin and increased transepidermal water loss. At this time, it is not known if a defective lipid barrier is present in atopic dogs. Normal and atopic canine skin were postfixed in ruthenium tetroxide and studied using transmission electron microscopy to determine structural differences within stratum corneum lipids. Intercellular lipid lamellae were graded on a semiquantitative scale. The deposition of stratum corneum lipid lamellae in atopic canine skin appeared markedly heterogeneous compared with that seen in normal canine skin. When present, the lamellae often exhibited an abnormal structure. The continuity and thickness of the intercellular lipid lamellae were significantly less in nonlesional atopic than in normal canine skin. These preliminary observations suggest that the epidermal lipid barrier is defective in atopic canine skin. Additional studies are needed to further characterize the biochemical defect and to possibly correct it with nutritional and/or pharmacologic intervention. PMID- 11732810 TI - Morbilliviral dermatitis in seals. AB - A juvenile female hooded seal (Cystophora cristata) and a juvenile male harp seal (Phoca groenlandica) stranded separately on the New Jersey (USA) coast and were taken to a marine mammal rehabilitation center. Both were lethargic and emaciated, had dermatitis, and died. Histologic skin lesions in the seals were similar and consisted of epidermal and follicular epithelial hyperplasia, hyperkeratosis, degeneration, and necrosis. The most distinctive finding was extensive syncytial zones bounded superficially by hyperkeratosis and deeply by hyperplastic basal cells. Eosinophilic intracytoplasmic inclusion bodies were present in epithelial cells. Morbilliviral antigen was demonstrated in the skin lesions by immunohistochemistry. Phocine distemper virus was detected in the skin by reverse transcription polymerase chain reaction and a phocine distemper virus specific probe using the Southern blot technique. This is the first report of morbilliviral dermatitis in marine mammals. PMID- 11732811 TI - Uterine angiolipoleiomyoma in a dog. AB - An 11-year-old crossbred Pomeranian bitch displayed a large intramural, well delineated uterine mass in one horn, near the junction with the uterine body. The mass was composed largely of mature adipose tissue, smooth muscle cells singly and in small clusters, anomalous medium-size and large arteries, and multifocal islands of cartilaginous and osseous tissues. Smooth muscle cells stained positively for desmin, and adipocytes and chondrocytes were positive for S-100 protein. This tumor has histologic and immunohistochemical features compatible with human uterine angiolipoleiomyoma, a rare tumor that has never been reported in the veterinary literature. This benign tumor is believed to be of a choristomatous nature. PMID- 11732812 TI - Teratoma in a feline unilateral cryptorchid testis. AB - A 14- x 12- x 6-cm (approximately 600 g) cryptorchid testis was surgically removed with the omentum from the posterior portion of the abdominal cavity of a 2-year-old male American Shorthair cat. At the time of a previous orchiectomy, a unilateral testis had been discovered in the scrotum. The cut surface of the cryptorchid testis consisted of solid areas with many cysts containing fluid and spongy soft areas. Histologically, the cryptorchid testis was composed of neuroectodermal components and diffuse immature glial tissues, cystic, tubular, and papillary epithelial tissues, and immature cartilaginous tissues. A metastatic mass in the omentum had histologic structures similar to those of the cryptorchid testis. The present case was diagnosed as teratoma in a feline unilateral cryptorchid testis. PMID- 11732813 TI - Medical applications of electronic nose technology: review of current status. AB - Electronic noses, or artificial sensors of odorants. have been developed over the last ten years to perform a variety of identification tasks in various industries. This powerful technology is only beginning to be introduced in the field of medicine, but is promising in its potential to assist in diagnosis. This article reviews electronic nose technology and some initial investigations of potential applicability of the technology in the field of medicine. PMID- 11732814 TI - Cold-induced rhinitis in skiers--clinical aspects and treatment with ipratropium bromide nasal spray: a randomized controlled trial. AB - Cold-induced rhinitis (CIR) is common among skiers and is perceived as a troublesome disease. We studied the clinical characteristics of CIR in a population of skiers and we evaluated the effectiveness of ipratropium bromide nasal spray (IBNS) in relieving symptoms in a double-blind placebo-controlled fashion. By means of specific questionnaires, we evaluated 144 subjects (69% men; mean age, 42.2 years). The prevalence of CIR was 48.6% and the distinctive symptom was rhinorrhea (96%), often severe. The prevalence of atopy was higher in the CIR patients (chi2; p = 0.004). Twenty-eight CIR subjects participated in a double-blind placebo-controlled cross-over trial for evaluating the effectiveness of IBNS (80 microg twice per day [b.i.d.]). The severity of symptoms was assessed by a visual analog scale, and the number of cleaning tissues used also was evaluated. The actively treated group showed a significant improvement of rhinorrhea (p = 0.0007) and a reduction in the number of cleaning tissues used (p = 0.0023). Only four mild local side effects were reported. We conclude that IBNS could be regarded as an optimal therapeutic option for treating CIR symptoms in skiers. PMID- 11732815 TI - Use of a portable spirometer for studies on the nasal cycle. AB - Studies based on the nasal cycle are difficult because multiple measurements of nasal patency need to be made over many hours. There is a great need for a simple portable instrument that can be used away from the clinical laboratory for studies on the nasal cycle. Our aim was to investigate the usefulness of a small portable spirometer in studying the nasal cycle. The Mir Spirobank spirometer was fitted with a nasal adapter to measure the volume of air expired from each nasal passage during a slow vital capacity (VC). The spirometer was used to measure the fractions of the slow VC volume of air expired through the right and left nasal passage in turn. Hourly measurements were made over a 5-hour period in six healthy volunteers. The spirometry measurements of the volume of air expired from each nasal passage were compared with nasal conductance of each nasal passage (airflow at 75 Pa) obtained by posterior rhinomanometry. The spirometer was found easy to use by both the investigator and the patients. Simple regression analysis of the spirometer and rhinomanometer measures of airflow partitioning found a correlation coefficient of r = 0.827 (p < 0.0001: n = 36). These results indicate that the partitioning of nasal expired volume measured by spirometry is directly comparable with partitioning of nasal airflow obtained with rhinomanometry. Spirometry has considerable advantages over rhinomanometry for studies on the nasal cycle because of portability and ease of use. PMID- 11732816 TI - Role of nasal valve in the surgically corrected nasal respiratory obstruction: evaluation through rhinomanometry. AB - The aim of this study was to evaluate the improvement of nasal flow and the fall of nasal resistance in 50 patients that underwent rhinoseptoplasty in our department and discuss the relative importance of valvular and septal deformities in nasal airway obstruction. Fifty consecutive patients underwent rhinoseptoplasty to improve nasal obstruction caused by severe septal deviation, external or internal valvular incompetence, or any combination of the three. We excluded patients with minor septal curvatures, septal perforations, or turbinate hypertrophy. Preoperative and postoperative rhinomanometry was performed on all 50 patients. In all 50 patients, septal and/or valvular surgery lowered nasal resistance in 90% of cases. Septoplasty alone with medial and basal osteotomies did not improve nasal flow (p < 0.4), whereas the correction of valvular obstruction alone increased nasal airflow in a statistically significant way (p < 0.0001). Moreover, patients with both valvular incompetence and septal deviation represented the group in which the greatest preoperative obstruction and the greatest postoperative improvement occurred. Nasal valvular function should be assessed with rhinomanometry in all preoperative rhinoplasty patients with airway obstruction. In many cases, valvular effects may surpass septal deviation as the primary cause of nasal airflow obstruction. PMID- 11732817 TI - Treatment of hypoxemia in obstructive sleep apnea. AB - Many patients suffering from obstructive sleep apnea (OSA) have intermittent oxygen desaturation associated with periods of apnea or hypopnea. Oxygen saturation levels below 90% are considered harmful. Usually, treatment is directed at correcting the apnea, which will in turn prevent hypoxemia. Unfortunately, many patients fail or are not candidates for nasal continuous positive airway pressure (CPAP) or surgical correction of their OSA. Forty-three patients with persistent OSA and nocturnal hypoxemia below 90% who were not candidates for additional surgical or CPAP therapy were treated with nocturnal oxygen supplementation. Standard symptoms associated with OSA and the Epworth Sleepiness Scale (ESS) were recorded before treatment and 30 days after the start of the treatment. In 21 patients, polysomnography studies were performed to compare the Respiratory Disturbance Index (RDI) score and minimum oxygen saturation levels when the patients were breathing room air or breathing 4 L/minute of oxygen by nasal cannula. Subjective symptoms of obstructive sleep apnea improved, and the ESS score significantly decreased after a 30-night treatment with oxygen. Split-night polysomnography showed a significant increase in minimum oxygen saturation during oxygen administration. The RDI did not significantly change with treatment. Oxygen administration for the correction of OSA-related nocturnal hypoxemia was both safe and effective in alleviation of OSA related symptoms. It also appeared to have a beneficial effect on minimum oxygen saturation levels. Thus, oxygen therapy may be considered a treatment option in patients who fail to comply with CPAP and are not candidates for a surgical procedure. PMID- 11732818 TI - Postoperative antibiotic use of septoplasty: a survey of practice habits of the membership of the American Rhinologic Society. AB - Prophylactic antibiotics are used routinely in otolaryngology. Little objective data exist concerning their use in sinonasal procedures. We wished to determine the practice patterns of otolaryngologists regarding prophylactic antibiotics for septoplasty patients. A survey of 743 (60.3% responded) members of the American Rhinologic Society was used to get information about their practice habits and rationale for using antibiotics after septoplasty. Sixty-six percent of respondents routinely used antibiotics. Postoperative infection and toxic shock syndrome (TSS) were the primary indications. Forty-four percent of respondents used some form of packing and 38% used a splint. There was no relationship between the annual number of procedures and the reasons for using antibiotics. Doctors who performed more than 50 cases a year were less likely to use packing or splints (p < 0.001). Older surgeons used less younger surgeons a packing/splints (p < 0.001). Respondents who used packing/ splints were more likely to use antibiotics (p = 0.008). Older respondents used packing whereas younger respondents used splints (p = 0.002). The rationale for antibiotics varied by age; re more concerned with TSS and older physicians were more concerned with infection (p = 0.005). This survey shows great variability in the usage and rationale for antibiotics after septoplasty. PMID- 11732819 TI - Antrochoanal polyps in children. AB - Although relatively rare, antrochoanal polyps represent one of the most common types of polyp diagnosed in children without cystic fibrosis. In an attempt to better define this entity and discuss treatment options, the histories and operative reports of all 25 children (aged 17 years and younger) diagnosed with an antrochoanal polyp between 1970 and 1997 at our institution were reviewed. All 25 children complained of nasal obstruction on presentation; other presenting symptoms included rhinorrhea (48%), snoring (36%), and mouth breathing (32%). All 25 patients were noted to have a mass in the nose on examination, and 16 (64%) also had a mass noted in the nasopharynx. All but 1 patient underwent surgical removal of the polyp: intranasal avulsion only, 2 patients; Caldwell-Luc procedures, 10 patients; intranasal procedures, 8 patients; and endoscopic procedures, 4 patients. Mean time to first recurrence was 44.5 months. Seven patients (29%) who underwent excision at our institution experienced recurrence, 3 after endoscopic procedures and 4 after intranasal procedures (with or without Caldwell-Luc; 1 of these patients had a second recurrence). Complications were unusual and included bleeding after pack removal (8.3%) and facial paresthesias (10%). Follow-up ranged from 2 days to almost 27 years and was aided by telephone interviews. We conclude that surgical treatment of these lesions is safe and effective. Endoscopic removal may result in a higher recurrence rate. PMID- 11732820 TI - Treatment of rhinocerebral mucormycosis by combination of endoscopic sinus debridement and amphotericin B. AB - Rhinocerebral mucormycosis (RCM) is an aggressive fungal infection with a high mortality rate. It frequently develops in patients with uncontrolled diabetes mellitus or immunocompromised patients. RCM typically presents in a rapidly fulminant manner with headache, fever, mucosal necrosis, and ophthalmic symptoms. Although the definitive diagnosis is achieved by histopathological examination, computed tomography (CT) scanning and magnetic resonance (MR) are the best imaging procedures in early diagnosis to assess the extent of the disease. PMID- 11732821 TI - Endoscopic repair of cerebrospinal fluid rhinorrhea: learning from our failures. AB - Endoscopic repair of cerebrospinal fluid (CSF) rhinorrhea is becoming a common procedure. The purpose of this study was to perform a literature analysis centering cases of treatment failure and to review our 31 cases with a 1-year minimum follow-up. An extensive search of the literature was conducted, which focused on success rate, follow-up, diagnostic techniques, graft material used, failure rate, and comments on failures. A retrospective analysis of our 31 patients was carried out, and all cases were treated with the endoscopic approach with a 1-year minimum follow-up. From the literature analysis, the median success rate at the first endoscopic attempt is 90%. Our success rate was 87.1%. Failures were analyzed. A unique protocol for CSF leak diagnosis does not exist; we suggest our diagnostic algorithm. Graft material used depends on the authors' experience, and based on this review of cases to date, did not significantly influence the success rate. The analysis of cases of failure shows that the majority of authors omit details. More research is needed to improve prevention of failures. PMID- 11732822 TI - The association between sinusitis and survival among individuals infected with the human immunodeficiency virus. AB - The purpose of this study was to determine the association between sinusitis and survival among human immunodeficiency virus (HIV)-infected persons. All patients enrolled in the adult spectrum of disease data base from November 1, 1990 to November 1, 1999 were included. Patients were followed until death, loss to follow-up, or the end of the study on January 10, 2000. A Cox proportional hazard regression analysis was conducted to evaluate the association between sinusitis, various other cofactors, and survival. Of the 7513 HIV-infected patients followed, 57% were <35 years old, 59.5% were black, 78.5% were male, and 20.8% had an opportunistic infection (OI) at entry. The incidence of one or more diagnoses of sinusitis in the cohort was 14.5%. The mean entry CD4 count for the entire cohort was 347.8 (SD, 298.9) and the mean follow-up time was 33.2 months (SD, 25.7). The mean CD4 count at the time of sinusitis diagnosis was 391 (SD, 316). In the multivariate analysis, older age and lower CD4 cell count were associated with death. Sinusitis, gender, and race were not associated with survival. Sinusitis is frequent in individuals infected with HIV. After adjusting for level of immunodeficiency, age, gender, and race, sinusitis is not associated with an increased hazard of death. This may have implications for treatment, because a diagnosis of sinusitis does not portend a poor prognosis in individuals infected with HIV. PMID- 11732823 TI - Differentiation-related p53 protein expression in nondysplastic sinonasal inverted papillomas. AB - To clarify p53 protein expression in nondysplastic sinonasal inverted papillomas, archived surgical specimens from 19 patients were studied using immunohistochemistry. Staining results were compared between inverted papillomas and adjacent, nonpapillomatous nasal mucosa. Further, immunoreactivity was compared between columnar (respiratory), transitional (cuboidal), and squamous epithelium in inverted papillomas. Positive staining was found in 17 of 19 inverted papillomas (89%). Immunoreactivity involved predominantly basal and parabasal cells and was either comparable or higher in inverted papillomas compared with adjacent mucosa. In 65% of immunoreactive inverted papillomas comparable staining results were seen between columnar (respiratory), transitional (cuboidal), and squamous epithelium. In 35% of p53 protein-positive inverted papillomas, enhanced immunoreactivity was observed in transitional (cuboidal) and squamous epithelium compared with columnar (respiratory) epithelium. Within these cases, immunoreactivity was either comparable or higher in squamous compared with transitional (cuboidal) epithelium. Conclusively, the expression of p53 protein is present in 89% of nondysplastic sinonasal inverted papillomas and also involves the adjacent, nonpapillomatous nasal mucosa. A tendency toward increasing p53 protein expression from nonpapillomatous nasal mucosa to inverted papilloma as well as along the metaplastic process from columnar (respiratory) to transitional (cuboidal) and finally squamous epithelium within inverted papillomas can be postulated. PMID- 11732824 TI - The effects of tibial rotation on the patellofemoral joint: assessment of the changes in in situ strain in the peripatellar retinaculum and the patellofemoral contact pressures and areas. AB - The objective of this study was to determine the effects of tibial rotation on in situ strain in the peripatellar retinaculum and patellofemoral contact pressures and areas. Patellofemoral joint biomechanics demonstrate a strong correlation with the etiology of patellofemoral disorders, such as chondromalacia, and are significantly influenced by tibial rotation. Six human cadaveric knees were used along with a patellofemoral joint testing jig that permits physiological loading of the knee extensor muscles. Patellofemoral contact pressures and areas were measured with a Fuji pressure-sensitive film, and the changes in in situ strain in the peripatellar retinaculum were measured with four differential variable reluctance transducers. Tibial rotation had a significant effect on patellofemoral joint biomechanics. The data showed an inverse relationship between increasing knee flexion angles and the change in patellofemoral contact pressures and in situ strain with tibial rotation. At higher knee flexion angles, the patella is well-seated in the trochlear groove and the function of the peripatellar retinaculum is minimized and less affected by tibial rotations. PMID- 11732825 TI - Design and validation of an instrument package designed to increase the reliability of ankle range of motion measurements. AB - OBJECTIVE: To validate a novel device termed the equinometer, which is designed to accurately measure ankle dorsiflexion. DESIGN: Test retest reliability analysis using serial measurements of dorsiflexion endpoint in a group of normal individuals. SETTING: Motion analysis laboratory. PARTICIPANTS: Ten healthy individuals. PRIMARY OUTCOME MEASURE: The mean and standard deviation of the absolute difference in dorsiflexion endpoint for the group. RESULTS: The mean absolute change in dorsiflexion endpoint for the group was 0.45 degrees with a standard deviation of 0.43 degrees. CONCLUSIONS: With the use of the device described, the mean change in dorsiflexion endpoint was well within acceptable clinical limits. The reliability of measurements obtained with the equinometer exceeds that which has been published with other techniques and devices. PMID- 11732826 TI - Microstructural characteristics of human skin subjected to static versus cyclic pressures. AB - Pressure ulcers are generally associated with external pressures exceeding internal capillary pressures over bony prominences when the body cannot initiate motor movement to change positions. This investigation evaluated microstructural changes occurring in human skin, in vitro, exposed to static versus cyclic pressures, simulating those recorded for heels of human subjects on various pressure-relief mattresses. Morphological data are reported for tissues exposed to pressure in a bench-scale loading system. Dynamic (cyclic-relief) pressure induced parallel alignments of connective tissue collagen bundles, which themselves became differentially oriented to various degrees perpendicular to the surface of the tissue. Static pressure, with no relief, invariably produced alignment of the collagen bundles of the connective tissue parallel to both one another and to the compressed tissue surface. The precursor to pressure ulcer formation may be microstructural alignment in response to the pressure conditions on tissue. PMID- 11732827 TI - State-of-the-art methods for geometric and biomechanical assessments of residual limbs: a review. AB - A review is presented on the state-of-the-art methods for geometric and biomechanical assessments of residual limb tissues. Residual limb assessments are needed throughout different stages of prosthetic management, namely preamputation, residual limb maturation, prosthetic design, prosthetic fitting, and subsequent follow-ups. Geometric assessments include the monitoring of the volumetric change as the residual limb matures after amputation. Geometric and biomechanical assessments of the residual limb are important considerations in prosthetic design. Quantitative geometric and biomechanical assessments are becoming more important in the development of a computer-aided system for prosthetic socket design. It is noted that, except for the external shape measurements of residual limbs, most of the instruments used in residual limb assessment are not particularly cost-effective for clinical applications in prosthetics. Further developments are needed to facilitate the incorporation of the internal geometric assessments and biomechanical assessments into the current computer-aided design systems for clinical prosthetics. PMID- 11732828 TI - Influence of training on biomechanics of wheelchair propulsion. AB - A quasi-experimental longitudinal design was used to compare pre- and posttraining biomechanical and physiological characteristics of wheelchair propulsion in manual wheelchair users (MWCU) across fresh and fatigue states. An instrumented wheelchair ergometer, 3D motion analysis, and computerized open circuit spirometry were used to collect joint kinetics and kinematics, handrim kinetics, propulsion temporal characteristics, and oxygen uptake pre- and posttraining during a submaximal exercise test to exhaustion. Each subject (n = 19) participated in a specific intervention program of supervised therapeutic exercise (strengthening, stretching, and aerobic exercise) for 6 weeks. Pre- and posttraining measurements were compared with the use of ANOVA with repeated measures. Significant training effects included increased exercise loads for all strengthening activities, decreased stroke frequency, increased maximum elbow extension angle, increased trunk and shoulder flexion/extension range of motion (ROM), increased handrim propulsive moment, increased wrist extension moment, and increased power output. Results suggest that this training program increased biomechanical economy (as defined by propulsive moment) without increasing shoulder or elbow joint stresses. PMID- 11732829 TI - Performance of an intramuscular electrode during functional neuromuscular stimulation for gait training post stroke. AB - The goal of rehabilitation for stroke patients in this research was to improve the volitional coordination of the swing phase and stance phases of gait. Functional neuromuscular stimulation (FNS) is a promising rehabilitation tool for restoring motor control. For our gait training protocols, FNS systems with surface electrodes were impractical. For the rehabilitation protocols that we defined, available implantable electrode designs did not meet desired criteria regarding fracture rate, invasiveness of placement procedures, and maintenance of position at the motor point. The criteria for the new intramuscular (IM) electrode design included minimally invasive electrode placement technique, accurate placement of electrodes, good muscle selectivity, consistency of muscle activation, good position maintenance of the electrode at the motor point, comfortable stimulus, and practical donning time for the system. A percutaneous electrode was designed for placement beneath the skin at the motor point of seven paralyzed or paretic muscles in the lower limb. A single-helical coil lead, a double-helical coil electrode, and fine wire barbs were design features that enhanced the anchoring capability of the electrode. A polypropylene core enhanced electrode durability. Implantation tools were custom-designed to enable accurate electrode placement without incision. We studied 17 subjects with a total of 124 electrodes. With the use of IM electrodes, FNS was provided for 1,413.8 electrode months. During this time, no instances of infection occurred. The measure of electrode integrity showed a 99% electrode survival rate. Throughout the treatment protocols, 93% of the electrodes delivered a good muscle response; 7% (nine electrodes) moved from the motor point and delivered a poor muscle response during the treatment protocol. Anchoring performance was higher for electrodes implanted in muscles that moved the hip (96.0%) and ankle joints (97.45%) compared with electrodes implanted in muscles that moved the knee joint (88.5%). Ninety-seven percent of the electrodes delivered a comfortable stimulus. Three percent delivered a stimulus that was uncomfortable at therapeutic levels and therefore were not used. We achieved gains in subject impairment and disability measures. The system proved to be practical for use in both clinical and home environments. PMID- 11732830 TI - A reusable, self-adhesive electrode for intraoperative stimulation in the lower limbs. AB - A suction-based stimulating electrode was designed and fabricated to allow intraoperative testing of lower-limb muscles during routinely scheduled surgical procedures. The suction device can adhere to a small exposure of muscle surface with reproducible contact forces and can maintain its geometric relationship to the underlying tissue for sufficient time to grade the resulting muscle contraction before removal and repositioning. When operated with a 10-cc syringe, the device can generate between 0 and 23 N of contact force; correlation between measured contact forces and those analytically predicted was 0.989. Preliminary animal testing indicates that the reusable device maintains its position over the nerve entry point even during vigorous active contractions of the stimulated muscle. Thus, it may be a valuable useful tool for locating the optimal site for a permanent electrode for functional electrical stimulation (FES) applications, as well as an ideal means of providing accurate and repeatable stimulation in various locations. PMID- 11732831 TI - Selectivity of intramuscular stimulating electrodes in the lower limbs. AB - Intramuscular (IM) electrodes have been used safely and effectively for decades to activate paralyzed muscles in neuroprosthetic systems employing functional electrical stimulation (FES). However, the response to stimulation delivered by these and any type of electrode can be limited by a phenomenon known as spillover, in which the stimulus intended to produce a contraction in a particular muscle inadvertently activates another muscle, causes adverse sensation, or triggers undesired reflexes. The purpose of this retrospective study was to determine the selectivity of monopolar intramuscular stimulating electrodes implanted in the lower limbs of individuals with motor and sensory complete paraplegia secondary to spinal cord injury (SCI) and to catalog the most common electrode spillover patterns. The performance records of 602 electrodes from 10 subjects who participated in a program of standing and walking with FES in our laboratory over the past decade were examined. Sixty percent (358) of these electrodes were "stable" (i.e., stimulated responses were consistent during the first 6 months postimplant), and 32% of all stable electrodes (113) exhibited spillover as noted in clinical and laboratory records. Common spillover patterns for eight muscle groups were tabulated and analyzed in terms of their functional implications. The beneficial (activation of synergistic muscles) or deleterious (activation of compromising reflexes, antagonists, or adverse sensation) effects of spillover were highly context dependent, with several potentially useful spillover patterns in certain phases of gait becoming undesirable and limiting in others. Knowledge of the selectivity of intramuscular electrodes and the patterns of spillover they exhibit should guide surgeons and rehabilitationists installing lower-limb neuroprostheses during the implantation process and allow them to better predict the ultimate functional usefulness of the electrodes they choose. PMID- 11732832 TI - Effects of stimulated hip extension moment and position on upper-limb support forces during FNS-induced standing--a technical note. AB - This study explores the effects of active hip extension moment produced by electrical stimulation on the support forces the arms must exert through an assistive device during quiet erect standing with functional neuromuscular stimulation (FNS) in individuals with spinal cord injuries (SCI). A static sagittal plane biomechanical model of human standing was developed to predict the effects of stimulated hip extension moment and sagittal plane hip angle on the arm support necessary to maintain an upright posture. Two individuals with complete thoracic SCI were then tested while they stood with continuous stimulation to the knee and trunk extensors. The steady-state active extension moment exerted at the hip was varied by activating different combinations of hip extensor muscles with continuous stimulation while steady-state support forces applied to the arms and feet during standing were measured. The steady-state support forces imposed on the arms during quiet standing decrease with increased stimulated hip extension moment and are highly dependent upon hip flexion angle, as predicted by the biomechanical simulations. Experimentally, the combination of gluteus maximus and semimembranosus stimulation produced three times more steady state hip extension moment than did stimulation of the gluteus maximus and adductor magnus. This resulted in a ten-fold decrease in body weight supported on the arms. More vertical postures (smaller hip flexion angles) improve the effectiveness of the hip extensor muscles in reducing the support forces placed on the arms. A single Newton-meter of stimulated hip extension moment with the hips fixed at 5 degrees of flexion results in almost five times the reduction in arm support forces as with the hips at 20 degrees. To minimize the forces applied by the arms on an assistive device for support while standing with FNS, these preliminary results suggest that (1) efforts should be made to assume the most erect postures possible and (2) muscles and stimulation paradigms that maximize active hip extension moment should be chosen. PMID- 11732833 TI - Comparison of two computer-automated procedures for tinnitus pitch matching. AB - Clinical assessment of the perceptual characteristics of tinnitus usually includes an attempt to match the pitch of tinnitus to a pure tone. A standardized clinical protocol for tinnitus pitch matching does not yet exist, and there is a history of unsuccessful attempts to obtain such measures reliably. The present study was designed to evaluate new protocols for identifying the perceived pitch of tinnitus, with the objectives of reducing testing time and improving test retest reliability. Two protocols ("Octave" and "Binary") were developed, each of which was patterned after the testing procedure previously developed at the Oregon Tinnitus Clinic and used to assess thousands of tinnitus patients. Both protocols use computer-automation to conduct testing; the protocols differ according to their specific testing algorithms. Twenty subjects with nonfluctuating tinnitus were each tested over two sessions. Results of testing revealed that both protocols could obtain pitch matches within 20 to 25 min. Reliability of responses was good for some subjects but not others, and the Binary protocol generally provided more reliable results. PMID- 11732834 TI - Reliability of hearing thresholds: computer-automated testing with ER-4B Canal Phone earphones. AB - This study was conducted to document test-retest reliability of hearing thresholds using our computer-automated tinnitus matching technique and Etymotic ER-4B Canal Phone insert earphones. The research design involved repeated threshold measurements both within and between sessions, and testing to evaluate the potential effect of eartip removal and reinsertion. Twenty normal-hearing subjects were evaluated over two testing sessions with the use of a fully automated protocol for determining thresholds with 1-dB precision. Thresholds were first obtained at 0.5-16.0 kHz, in one-third octave frequency steps (16 test frequencies). The octave frequencies were then retested, first without removing the eartips, then after eartip removal and replacement. Responses between sessions differed by an average of 2.5 dB across all 16 test frequencies, and 91.5 percent of the repeated thresholds varied within +/-5 dB (98.1 percent within +/-10 dB). Reliability of within-sessions thresholds was also good, and there was no effect of eartip removal and replacement. PMID- 11732835 TI - Modified constraint induced therapy: a randomized feasibility and efficacy study. AB - This case series examined the feasibility and efficacy of a modified constraint induced therapy (CIT) protocol administered on an outpatient basis. The Fugl Meyer Assessment of Motor Recovery After Stroke (Fugl), Action Research Arm Test (ARA), Wolf Motor Function Test (WMFT), and Motor Activity Log (MAL) were administered to six patients between 2 and 6 months poststroke (CVA) exhibiting stable motor deficits and learned nonuse of the affected limb. Two patients then participated in half-hour physical and occupational therapy sessions three times/week for 10 weeks. During the same period, their unaffected arms and hands were restrained 5 days/week during 5 hours identified as times of frequent use. Two other patients received regular therapy and two control patients received no therapy. The ARA, Fugl, WMFT, and MAL were again administered after 10 weeks. Patients receiving modified CIT exhibited substantial improvements on the Fugl, ARA, and WMFT, as well as increases in amount and quality of use of the limb using the MAL. Patients receiving traditional or no therapy exhibited no improvements. Results suggest that modified CIT may be an efficacious method of improving function and use of the affected arms of patients exhibiting learned nonuse. PMID- 11732837 TI - Guest editorial: aesthetics of walking. PMID- 11732836 TI - Pulmonary function testing in spinal cord injury: effects of abdominal muscle stimulation. AB - The purpose of this study was to assess the effects of applying transcutaneous electrical stimulation to paralyzed abdominal muscles during pulmonary function testing (PFT) of individuals with spinal cord injury (SCI). Ten male subjects with anatomical level of SCI between C5-T7 were studied. Subjects performed PFTs with and without electrical stimulation delivered to the abdominal muscles. Subjects with the lowest percentage of predicted expiratory volumes and flows demonstrated the greatest improvement when electrical stimulation was delivered during forced expiration. The overall increases seen in percent of predicted for the study sample were 23 percent for forced vital capacity (FVC), 16 percent for forced expiratory flow in 1 s (FEV1), and 22 percent for peak expiratory flow rate (PEF). Contractions of paralyzed expiratory muscles in response to electrical stimulation during the performance of PFT maneuvers can significantly improve FVC, FEV1, and PEF in some individuals with SCI. PMID- 11732838 TI - Amphibian gastrulation: history and evolution of a 125 year-old concept. AB - The hypothetical gastraea concept, proposed by Haeckel (1874) to be an ancestral form common to all Metazoans, relied on the characterization of a gastrula stage in their embryonic development. The first steps that led to this characterization in Amphibian embryos fell into oblivion and deserve mention. Similarly, controversial debates about gastrula formation from the blastula, about simultaneous appearance of the three germ layers as opposed to a theoretical diploblastic embryo and about the occurrence of inward morphogenetic cell movements versus that of delamination processes, lasted for years. Following a half-century of polemic (1875-1925), Vogt's studies clearly and definitively established the reality and the complexity of morphogenetic movements, but this breakthrough long remained without further consequences. Holtfreter (1943,1944) illuminated unknown aspects of living gastrula cells and his observations helped to define many problems to be solved. During the second half of the 20th century, cell and molecular biology techniques, applied to the study of cell-cell and cell matrix interactions, have brought new insights into the mechanisms of gastrula cell movements. Gene expression during these phenomena still remains an open question, as shown by a few recent studies: this situation strikingly contrasts with the many achievements that have been accomplished during the last decade in the analysis of induction phenomena during gastrulation. PMID- 11732839 TI - Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. AB - Events after fertilisation have been carefully studied in the last decades. However, there are still several questions to be clarified in relation to the signalling pathway initiated by the sperm, the identification of proteins or factors involved in the activation of the arrested oocyte, and the inactivation of specific molecules involved in the meiotic arrest. The present state of knowledge in mammalian fertilisation allows the development of activation protocols that closely mimic the events initiated by the sperm according to certain major factors (MPF activity and MAPk activity). These protocols are successfully used for the activation of oocytes after NT giving rise to offspring. Few cloned animals have yet been produced. However, the pregnancy and the survival rates after birth are not significantly different when different activation protocols are compared. This fact argues fora major reason forthe low success in the efficiency of NT. Eventually, factors related to the recipient oocyte, the donor cell or the culture conditions are part of these major problems that the reconstructed embryo has to overcome to develop into a normal offspring. Nonetheless, the development of activation protocols that closely imitate the mechanism of activation initiated by the sperm are of special interest to improve the developmental potential of cloned embryos. PMID- 11732840 TI - How many blastomeres of the 4-cell embryo contribute cells to the mouse body? AB - The aim of this study was to estimate how many blastomeres of the 4-cell mouse embryo contribute cells to the embryo proper and finally to the animal. To this end, 4-cell embryos of pigmented and albino genotypes were disaggregated and single blastomeres (henceforth called '1/4' or 'quarter' blastomeres) were reaggregated in the following combinations: one 'pigmented' blastomere + three 'albino' blastomeres or vice versa (henceforth called '1+3') and two pigmented blastomeres + two albino blastomeres (henceforth called '2+2'). The aggregations were cultured in vitro and transferred as blastocysts either to the oviduct or uterus of pseudopregnant females. Recipients were allowed to litter naturally, or the foetuses were removed by Caesarian section and raised by lactating foster mothers. Chimaerism was assessed on the basis of coat (adults) or eye pigmentation (dead neonates). Among 28 '1+3' animals, there were 13 chimaeric and 15 non-chimaeric individuals. The pigmentation of non-chimaeras was always concordant with the genotype of the three 1/4 blastomeres and not with the genotype of the single blastomere in the given aggregation. These results make rather unlikely the possibility that the mouse is built of cells derived either from one or all four 1/4 blastomeres. Both two remaining options (2 or 3 1/4 blastomeres) are conceivable but the observed ratio of chimaeras to non-chimaeras among '1+3' animals (13:15) fits better the assumption of two 1/4 blastomeres contributing cells to the animal body. This assumption finds additional support in the observation that among '2+2' animals there were non-chimaeras (5 out of 7) and these would not have been expected should three 1/4 blastomeres contribute cells to the mouse body. PMID- 11732842 TI - Localization of type IV collagen a 1 to a 6 chains in basement membrane during mouse molar germ development. AB - The dental basement membrane (BM) putatively mediates epithelial-mesenchymal interactions during tooth morphogenesis and cytodifferentiation. Type IV collagen alpha chains, a major network-forming protein of the dental BM, was studied and results disclosed distinct expression patterns at different stages of mouse molar germ development. At the dental placode and bud stage, the BM of the oral epithelium expressed alpha 1, alpha 2, alpha 5 and alpha 6 chains while the gubernaculum dentis, in addition to the above four chains, also expressed a 4 chain. An asymmetrical expression for alpha 4, alpha 5 and alpha 6 chains was observed at the bud stage. At the early bell stage, the BM associated with the inner enamel epithelium (IEE) of molar germ expressed alpha 1, alpha 2 and alpha 4 chains while the BM of the outer enamel epithelium (OEE) expressed only alpha 1 and a 2 chains. With the onset of dentinogenesis, the collagen a chain profile of the IEE BM gradually disappeared. Howeverfrom the early to late bell stage, the gubernaculum dentis consistently expressed alpha 1, alpha 2, alpha 5 and a 6 chains resembling fetal oral mucosa. These findings suggest that stage- and position-specific distribution of type IV collagen alpha subunits occur during molar germ development and that these changes are essential for molar morphogenesis and cytodifferentiation. PMID- 11732841 TI - Isolation and characterization of a Xenopus gene (XMLP) encoding a MARCKS-like protein. AB - We have identified a cDNA coding for a Xenopus MARCKS-like protein (XMLP) from a cDNA library prepared from activin-treated ectoderm. Using whole-mount in situ hybridization and RT-PCR, we found XMLP maternal transcripts during the cleavage stages. After MBT, the signals were restricted to the neural plate. Subsequently XMLP was expressed predominantly in the brain, somites and pronephros. Ectopic expression of XMLP resulted in eye and axis defects and in a change of the expression pattern of Krox 20, a neural marker for rhombomeres 3 and 5. Injected XMLP caused apoptosis. It was characterized by loss of intercellular adhesion contacts, transient plasma membrane ruffling at gastrula, and epithelial disruption attailbud stage. Overexpression of mutant XMLPs showed that this phenotype was correlated with its putative PSD domain and glycine at position 2. The embryos injected with a morpholino oligo complementaryto XMLPmRNA showed malformations of the anterior axis and eye defects. Extirpation experiments indicated that the phenotypes might be correlated with disturbed morphorgenetic movements rather than an inhibition of induction process. Overexpression of XCYP26 resulted in a shift of the expression pattern of XMLP. In the early tailbud stage (stage 20) the signal stripe in the XCYP26 injected half of the embryo got diffuse or even disappeared. This observation suggests that retinoic acid plays an important role in the regulation of XMLP. Our results suggest that XMLP might participate in pattern formation of the embryonic axis and the central nervous system. PMID- 11732843 TI - A hormone-dependent post-translationally regulated mutant for investigating type I cadherin function. AB - Type I cadherins are Ca2+-dependent cell adhesion molecules. Their function in early Xenopus laevis development has been extensively studied in recent years, by injecting synthetic mRNAs encoding dominant negative mutants with deletions of the extracellular domain into embryos. However, studies at post-gastrula stages have been hampered by the inabilityto progress through post-gastrula development in embryos expressing these mutant proteins. This problem has been partly overcome by injecting into a few targeted blastomeres in stage 6 N.F. embryos, but only restricted studies are possible with this technique. Several studies have made use of the hormone-binding domain (HBD), which is activated by hormones. In this study, we used this method to analyze the activity of dominant negative cadherins. We generated a mutant E-cadherin (AE-Cad, consisting of the cytoplasmic domain and transmembrane domain) fused to the hormone-binding domain of estradiol receptor (HBDER) and we validated this technique with functional analyses. The function of the mutant deltaE-HBDER was strictly dependent on hormone induction. This conditional mutant had the same effects and exerted the same dominant negative function as the corresponding constitutive mutant. PMID- 11732844 TI - NUDC expression during amphibian development. AB - To identify gene products important for gastrulation in the amphibian Pleurodeles waltl, a screen for regional differences in new protein expression at the early gastrula stage was performed. A 45 kDa protein whose synthesis was specific for progenitor endodermal cells was identified. Microsequencing and cDNA cloning showed that P45 is highly homologous to rat NUDC, a protein suggested to play a role in nuclear migration. Although PNUDC can be detected in all regions of the embryo, its de novo synthesis is tightly regulated spatially and temporally throughout oogenesis and embryonic development. New PNUDC synthesis in the progenitor endodermal cells depends on induction by the mesodermal cells in the gastrula. During development, PNUDC is localized in the egg cortical cytoplasm, at the cleavage furrow during the first embryonic division, around the nuclei and cortical regions of bottle cells in the gastrula, and at the basal region of polarized tissues in the developing embryo. These results show for the first time the expression and compartmentalization of PNUDC at distinct stages during amphibian development. PMID- 11732845 TI - Variability, heritability and genetic advance in pheno-morphic and agronomic traits of tef [Eragrostis tef (Zucc.) Trotter] germplasm from eight regions of Ethiopia. AB - One hundred and twenty tef Eragrostis tef (Zucc.) Trotter, germplasm lines from eight regions of Ethiopia were field evaluated in two randomized complete blocks at Debre Zeit and Melkassa during the 1995/96, and at Debre Zeit and Alem Tena during the 1996/97 main seasons. The objectives were to assess the extent and pattern of their diversity with respect to 17 pheno-morphic and agronomic traits, and to obtain an insight into the broad sense heritability (H) and genetic advance (GA) expectations of the various traits. Discernible (p < or = 0.05) variability among regions and among genotypes within regions were observed in seven and nine of the traits, respectively. In the combined analysis of variance over environments, genotypes showed substantial (p < or = 0.05) variation in all traits, and genotypes and environments interacted significantly on nine of the traits. The phenotypic and genotypic coefficients of variation varied in that order from about 3% for days to maturity to 28% for grain yield/plant, and from less than 2% for days to maturity to 15% for number of fertile tillers/plant. Estimates of H were lowest for shoot phytomass/plant (17%) and highest for panicle length (75%). The minimum GA expectations were for days to maturity (2%) and the maximum was for number of fertile tillers/plant (23%). About 67% of the gross phenotypic diversity in the germplasm was explained on the basis of four principal components (PCs). Up to 28% of the variance extracted by the first PC was due mainly to variations in the second and first basal culm internode diameters, culm and panicle length, days to panicle emergence, and number of panicle branches and fertile tillers/plant. Largely variations in the length of the two basal culm internodes and grain yield/plant contributed to 19% of the whole variance accounted for by the second PC. Generally, the study showed that there is ample genetic variation in the genotypes evaluated and this presents a valuable base for use in the improvement work. PMID- 11732846 TI - Sources of wheat powdery mildew resistance from wheat-rye and wheat-Leymus hybrids. AB - Introgression of germplasm from rye and Leymus mollis has been used in the objective of broadening mildew resistance in wheat. The mildew resistant wheat lines were isolated from crosses of three hexaploid triticale lines, a mildew resistant double wheat-rye substitution line and a mildew resistant hexaploid wheat-L. mollis hybrid with three mildew susceptible breadwheats. Three different mildew isolates with a broad virulence have been used and lines with different resistance patterns, including resistance to all three mildew isolates, were selected. From a total of about 13,000 lines, eight lines from the triticale crosses, 58 from the double wheat-rye substitution line crosses and six from the L. mollis-hybrid crosses showed homozygous resistance to all three isolates. A differential tester set of wheat lines with known mildew resistance genes including earlier known resistance genes from rye showed a similarity between IRS cultivars and the largest category in the triticale populations, indicating a higher frequency of IR chromosomes. The results are discussed in relation to the differential tester set and in relation to broadening the genetic base of resistance in wheat by interspecific hybridization. PMID- 11732847 TI - Cytogenetics of eight Neotropical species of Chauliognathus Henzt, 1830: implications on the ancestral karyotype in Cantharidae (Coleoptera). AB - Meiotic chromosomes of eight Neotropical species of the genus Chauliognathus occurring in Southern Brazil were analysed from testis cells. The meioformula of all the species was n = 6 + X (with six pairs of autosomes and one X chromosome). This meioformula is also present in C. scriptus plus two other species of other genera of Cantharidae, suggesting an evolutionary tendency towards the compaction of the karyotype starting from the ancestral Polyphagan meioformula n = 9 + Xy(p). The presence of an XO sex chromosome mechanism in all cantharid species studied to date indicates that the reduction of the number of autosomes was probably paralleled by the loss of the small typical Polyphagan yp heterosome. Thus the meioformula n = 6 + X seems to be an apomorphy for Cantharidae and may have evolved soon after the separation of the ancestral cantharid from the other families of Polyphaga. PMID- 11732848 TI - The isolation of a mutation causing abnormal cytokinesis in male and split chromocenter in female meiosis in Drosophila melanogaster. AB - The genetic screen for the meiotic mutations showing chromosome non-disjunction in mosaic clones of female germ line generated by FLP-FRT mediated mitotic recombination was performed. The sterile meiotic mutation ff16 (69D1-70A2) was found among the mutants obtained. In the male germ line the mutation showed lack of meiosis 1 cytokinesis and other meiotic abnormalities. The sterility of the mutant is due to the lack of the sperm motility. In female germ line the morphological defects-decreased number of ovarioles and nurse cells in the egg chambers is visible. At the cell level the mutation showed karyosome fragmentation constituting to the gene participation in chromocenter formation/maintance. The cases of the spindle fragmentation revealed the processes acting in female meiotic metaphase. Premeiotic and mitotic defects of the mutation have also been detected. PMID- 11732849 TI - Steep variation in mitochondrial DNA and B chromosomes among natural populations of Eyprepocnemis plorans (Acrididae). AB - Restriction enzyme analysis of the mitochondrial DNA of Eyprepocnemis plorans with 9 restriction enzymes revealed low variability--only EcoRI revealed any variability with 3 distinguishable digestion patterns, here named types I, II and III. The samples studied were collected from regions where different types of B chromosome exist. The demes from the central area show the B1 type (as a relict of a probably once continuous distribution) and also mtDNA type II. These show a parallel substitution towards the SW neighbouring demes by other Bs and mtDNA type I. However, the latter mtDNA type had a wider distribution and is predominant, or even the only one found in most other samples where other Bs exist. Considering the mtDNA as a marker unlinked with the Bs, some genetic differences should exist among areas defined by the B chromosomes. Our results support the hypothesis that in the central region the B chromosomes and the mtDNA are involved in two different events of substitution, but these do not necessarily occur in the same way. PMID- 11732850 TI - Physical mapping of 45S and 5S rDNA on maize metaphase and sorted chromosomes by FISH. AB - Physical locations of 45S and 5S rDNA were detected in maize by fluorescence in situ hybridization (FISH). The FISH results on metaphase chromosome spreads showed the 45S rDNA was located just at the nucleolus organizer region (NOR) on 6S (the short arm) of chromosome 6) as expected, and 5S rDNA at the distal region of 2L (the long arm of chromosome 2). No signals were detected on the other maize chromosomes for these two probes. The precision of the chromosomal position of a hybridization site is related to the number of measurements. We also mapped the location of 5S rDNA at the same sites on 2L by FISH on sorted chromosomes. We could calculate more precisely the percentage distance of hybridization signals since we had large numbers of target chromosomes on a small spot on the slide by flow sorting. The percentage distance from centromere to the hybridization site was 85% for 5S rDNA on 2L. The physical location of 5S rDNA was inconsistent with its genetic site, which was positioned in the middle of genetic linkage group 2. A FISH procedure for mapping genes on sorted plant chromosomes is described and occurrence of only one 45S or 5S rDNA site on maize genome is discussed. PMID- 11732851 TI - A case of trisomy 28 in cattle revealed by both banding and FISH-mapping techniques. AB - A case of presumed trisomy 22 in cattle, as earlier reported, was investigated by using both chromosome banding and FISH-mapping techniques. A one year old Hereford female calf showed slow growth, brachygnathia superior, hyper salivation. strabisbusmus convergence, macroclitoris and supernumerary nipples. At necropsy, a duplication of the uterine cervix and an extra aortic cusp were also observed. The R-banding technique revealed that BTA28 (not BTA22, as erroneously earlier reported) was involved in the trisomy. This was confirmed by FISH-mapping with chromosome-specific markers. Phenotypic characteristics achieved in cattle carrier and those earlier reported in humans carrying partial trisomy 10 were discussed. PMID- 11732852 TI - Genetic analysis of mosaicism in 53 women with Turner syndrome. AB - Mosaicism involving the sex chromosomes is a common finding in women with Turner syndrome (TS). It is especially important to detect Y-chromosomal material, since this is a risk factor for the development of gonadoblastoma. Recent studies have also indicated that the frequency of 45,X cells may be used to predict prognosis. As part of an ongoing multi-disciplinary study, we have examined the extent of Y chromosomal material and sex chromosomal mosaicism and its tissue specificity in 53 women with TS. The results of lymphocyte karyotyping were compared with the use of interphase X/Y fluorescence in situ hybridisation (FISH) analysis of lymphocytes and buccal mucosal cells. As could be expected, an extended FISH analysis detected more Y-chromosomal material than karyotyping (in 15% vs. 11% of the women, respectively) and also detected more X-chromosomal mosaicism among the TS women (in 70% vs. 45 % of the women, respectively). In half of the women, tissue-specific differences between lymphocytes and buccal mucosal cells were found. Based on these results, we suggest the use of X/Y interphase FISH as a complement to karyotyping in order to obtain a more complete knowledge of the chromosome constitution of each individual with TS. PMID- 11732853 TI - Clines maintained by overdominant selection in hybrid zones. AB - Hybrid zone models often consider environment-independent selection to operate against all hybrids. However, empirical studies suggest that hybrids may be as fit or fitter than the hybridizing parental taxa in some environments. In this study we develop a novel mathematical model to explore the effects of one form of hybrid superiority on the genetic structure of hybrid zones. Our primary goals were to investigate the allele frequency clines at a locus experiencing overdominant selection and at a linked neutral or underdominant locus. Our results indicate that overdominant selection results in flat equilibrium allele frequency clines throughout the hybrid zone and an excess of heterozygosity relative to Hardy-Weinberg equilibrium. However, the genetic clines at linked neutral or underdominant loci tend not to reflect this overdominance even when the loci are tightly linked. Overall, we conclude that overdominance is unlikely to be detected in genetic surveys unless many loci are assayed. PMID- 11732854 TI - Two postglacial immigration lineages of the polyploid Cerastium alpinum (Caryophyllaceae). AB - The plant cover of Fennoscandia is young because of the recent glaciation. This study covers the early stages of diversification and the genetic consequences of postglacial migration of a hermaphroditic polyploid plant. Cerastium alpinum. It has a continuous distribution in the alpine region, where it grows on alpine heaths and serpentine soils that are rich in heavy metals. Within the boreal forest C. alpinum has a scattered distribution on serpentine, dolomite and steep slopes. Plants from 31 populations in Norway, Sweden and Finland were subjected to enzyme electrophoresis. Analyses of the enzyme phenotypes suggest that C. alpinum has colonized Fennoscandia through two postglacial immigration events resulting in a southeastern and a southwestern lineage. These two lineages seem to meet in a hybrid zone in northern Sweden. Large genetic differences were found among most populations in both the southeastern and the southwestern lineages. This suggests that the populations are effectively isolated from each other. PMID- 11732855 TI - The nature of apomixis in cassava (Manihot esculentum, Crantz). AB - Apomixis in cassava is formed by aposporic embryo sacs which arise from cells in nucellus or from cells in the sexual embryo sac. There is an association between sterility and apomixis. It seems that sterility caused by defected meioses triggers a specific gene for exhibiting apomixis. This apomictic mechanism is favoured by natural selection since it offers an escape from lethality and provides a perpetuation of the current genotype. PMID- 11732856 TI - Transformation and regeneration capacities for five Nordic barley elite cultivars -evaluation of tissue culture response and transient expression. AB - Using both biolistic and Agrobacterium-mediated DNA delivery, we have investigated the transformation and regeneration capacity for five Nordic elite cultivars of barley. Transformation was followed as transient expression of the uidA or gfp gene in barley callus. Callus formation and regeneration of transformed callus were evaluated based on callus induction frequency, growth rate, callus appearance, and shoot formation frequency. From the accumulated results, one of the elite cultivars has been selected for our ongoing work in molecular breeding of barley. PMID- 11732857 TI - IL-1beta-induced expression of matrix metalloproteinases and gliostatin/platelet derived endothelial cell growth factor (GLS/PD-ECGF) in a chondrosarcoma cell line (OUMS-27). AB - The purpose of this study was to examine how chondrocytes are involved in the molecular mechanism of inflammation in rheumatoid arthritis (RA). A chondrosarcoma cell line (OUMS-27) was cultured and treated with interleukin 1beta (IL-1beta). Changes in the expression levels of matrix metalloproteinase-1 (MMP-1), metalloproteinase-13 (MMP-13), and gliostatin/platelet-derived endothelial cell growth factor (GLS/PD-ECGF) were assessed by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assays. IL-1beta induced the expressions of MMP-1, MMP-13, and GLS mRNAs and proteins in a dose-dependent manner. Selective inhibition of the p38 mitogen activated protein kinase (p38 MAPK) pathway with SB 203580 and SB 202190 blocked the expression of MMP-1, MMP-13, and GLS more strongly than selective in hibition of the extracellular signal-regulated kinase 1 and 2 (ERK1/2) pathway by PD 98059. These findings suggest that chondrocytes may intensify cartilage destruction and inflammation in RA by the induction of MMP-1, MMP-13, and GLS by IL-1beta and that the p38 MAPK pathway plays an important role in these inductions. PMID- 11732858 TI - Retroviral activity in Behcet's syndrome and systemic lupus erythematosus detected by a PCR-based reverse transcriptase assay. AB - The objective of this study was to assess retroviral activity in Behcet's syndrome (BS) and systemic lupus erythematosus (SLE) patients. Serum and peripheral blood mononuclear cells (PBMC) were obtained from patients and normal volunteers and PBMC cultured with and without phytohaemagglutinin stimulation. Reverse transcriptase (RT) activity in serum and culture supernatants was measured using a sensitive polymerase chain reaction-based assay. An RT activity above the levels in normal controls was detected in a minority of patients with BS (2/15) and SLE (1/13) and was typically present in all three types of sample. Elevated levels of RT activity were not detected in follow-up samples from the two BS patients. Our findings indicate that elevated RT activity is present in only a minority of patients with BS and SLE. Simultaneously elevated activity in all three sample types implicates PBMC as the source of this retroviral activity. PMID- 11732859 TI - Association of T102C polymorphism of the 5-HT2A receptor gene with psychiatric status in fibromyalgia syndrome. AB - Serotonin (5-HT) is a key neurotransmitter in the central nervous system. It is suggested that serotonergic dysfunction may be involved in the pathophysiology of fibromyalgia syndrome (FS). In this study, we aimed to investigate T102C polymorphism of the 5-HT2A receptor gene in FS. Fifty-eight patients with FS and 58 unrelated healthy volunteer controls were included in the study. In both groups, the C/C, C/T, and T/T genotypes of the 5-HT gene were represented in 31% (22.4% in controls), 50% (53.4%), and 19% (24.1%), respectively. The 5-HT2A receptor gene polymorphism results were not significantly different between patients and controls (chi squared test, P>0.05). There was a significant correlation between patients with the T/T genotype and the subgroup according to the SCL-90-R test, (analysis of variance, P<0.05). We also saw that patients with the T/T genotype had the lowest pain threshold. CONCLUSION. T102C polymorphism of the 5-HT2A receptor gene is not associated with the etiology of FS. Our results also indicate that the T/T genotype may be responsible for psychiatric symptoms of FS. PMID- 11732860 TI - Psychosocial function, clinical status, and radiographic findings in a group of chronic low back pain patients. AB - Low back pain is considered a problem with multiple facets for which the underlying causative factors should be determined. The aim of this study was to evaluate the relationships between depression, clinical status, and radiographic findings in a group of fifty patients with low back pain for more than 6 months. The patients underwent clinical examination and they completed Beck depression inventory (BDI), Aberdeen back pain scale (ABPS) and research questionnaire. Radiographic evaluations were performed. Clinical score and duration of symptoms were found to be positively correlated. The BDI scores were not found to be correlated with the existing variables. The ABPS scores were positively correlated with clinical scores and number of medications used. PMID- 11732861 TI - Comparative analysis of psoriatic spondyloarthropathy between men and women. AB - This study analyzed gender-related differences in a cohort of patients with psoriatic spondyloarthropathy (SpA) We performed a retrospective cross-sectional study of 100 patients (mean age 48 +/- 14 years; 63 men, 37 women), diagnosed as having psoriatic SpA on the basis of ESSG criteria. All patients were studied according to a standard protocol, and HLA-B27 and Cw status were analyzed in the study population and their frequencies compared to that of 177 healthy blood donors. The clinical features of PsSpA were compared between men and women by univariate analyses. Twenty-three patients showed isolated axial disease (M:F ratio 3.6:1), 36 had polyaxial disease (M:F ratio 1:1), and 41 showed oligoaxial pattern (M:F ratio 1.7:1). HLA-B27 was correlated with male sex (P=0.002) and isolated axial disease (P=0.016). Univariate analysis showed female sex to be correlated with lower complement levels (P < 0.05), erosive disease (P = 0.05), higher swelling joint count (P = 0.002), and higher scores on the Health Assessment Questionnaire-Specific for SpA (P < 0.05). The HLA-B27 antigen seems to be a defined genetic risk factor only in men with psoriatic SpA. The extent of the spondylitic process is quite similar between the two, although women show poorer functional performance and more aggressive peripheral disease. PMID- 11732862 TI - Serum interleukin-12, interleukin-15, soluble CD26, and adenosine deaminase in patients with rheumatoid arthritis. AB - CD26, a transmembrane ectoenzyme, is overexpressed on rheumatoid arthritis (RA) peripheral blood T cells. As it has been recently described that IL-12 and IL-15 upregulate CD26 in vitro, we hypothesized that this CD26 overexpression might be interleukin dependent. The concentrations of IL-12 and IL-15, and soluble CD26 and adenosine deaminase (enzymes related to CD26) were analyzed in the serum of 35 patients with active and inactive RA and of healthy control subjects. IL-12 and IL-15 levels were significantly higher in patients' serum, independently of disease activity, even in patients on steroid therapy, i.e., the present therapies cannot eradicate their origin. Soluble CD26 was significantly reduced and related to the disease activity. In particular, it correlated inversely with the number of swollen joints. Although these data did not support our hypothesis, they support that interleukins not only initiate RA pathology but they can also participate in the maintenance of this immune response. PMID- 11732863 TI - Association of inclusion body myositis with subacute cutaneous lupus erythematosus. AB - We present the case of a 71-year-old man with inclusion body myositis combined with subacute cutaneous lupus erythematosus and dysphagia. Although inclusion body myositis is usually resistant to immunosuppressive therapy, this patient improved under treatment with corticosteroids. The presented case is discussed in the context of earlier reports of inclusion body myositis and lupus erythematosus. PMID- 11732864 TI - A case of ochronosis: upper extremity involvement. AB - We present an ochronotic patient with spondylosis and upper extremity involvement. We also evaluated radiologic findings of joints that were involved and MRI features of the lumbar spine. PMID- 11732865 TI - Carpal tunnel syndrome in HIV-infected patients treated with highly active antiretroviral therapy: other case reports. AB - A possible association between carpal tunnel syndrome and HIV infection has been suggested only once until now. Two patients with HIV infection are described who presented confirmed carpal tunnel syndrome while on prolonged protease inhibitor containing highly active antiretroviral therapy (HAART). Their disease course and long-term outcome after medical and/or surgical interventions is presented and discussed according to available evidence from the literature. In our patients, carpal tunnel syndrome occurred in apparent absence of all presumed risk factors of this disease and metabolic abnormalities potentially related to HIV protease inhibitor administration. PMID- 11732866 TI - Administration of plasmid DNA expressing human interleukin-6 significantly improves thrombocytopoiesis in irradiated mice. AB - When people are exposed to large doses of ionizing rays in a short time, hematopoiesis is impaired and hemorrhage is one of the major clinical features. Suddenly decreasing platelet counts are responsible for the life-threatening hemorrhagic complication. Therefore, some cytokines have been used to improve thrombocytopoiesis in various radiation-induced thrombocytopenia models. Current measures for this purpose involve repeated intravenous or subcutaneous injections of recombinant proteins, which are expensive and inconvenient, or gene therapy with viral vectors that could not obviate the risk of infection. We tried to determine the possibility of gene therapy with plasmid vectors for radiation induced hematopoietic injury, which could overcome the above-mentioned problem. In this study, we describe the enhanced efficiency of radiation on gene transfer with plasmid vector in vivo and the physiological role of expressed human interleukin-6 (hIL-6) in vivo on a radiation-induced thrombocytopenia model. After a single intramuscular injection of plasmid hIL-6 DNA on 6.5-Gy-irradiated mice, the hIL-6 protein level in mouse plasma was determined with enzyme-linked immunosorbent assay (ELISA). The level of hIL-6 began to increase from the 4th day, reached the peak value on about the 11th day, and remained at a higher level on the 28th day. Meanwhile, unirradiated mice injected with the same amount of plasmid DNA showed less hIL-6 on the 11th day after administration. Further experiments demonstrated that the hIL-6 level in 7.5-Gy-irradiated mice was about three times higher than that of 5.0-Gy-irradiated mice, suggesting radiation could improve gene transfer efficiency of plasmid DNA in vivo and might be dependent on radiation doses. The expression of hIL-6 in vivo showed a significant effect on hematopoietic recovery after radiation. Not only the platelet nadir in peripheral blood, but also the number of colony-forming cells in bone marrow rose. The increased platelet counts were partially due to the increase of reticulated platelet that reflected the activity of a given population of megakaryocyte in bone marrow. We conclude that radiation could significantly enhance the gene transfer efficiency of plasmid DNA and that gene therapy with plasmid vectors for radiation-induced hematopoietic injury might be more effective than other diseases without DNA repair. PMID- 11732867 TI - Abnormal platelet adhesion on abdominal vessels in asymptomatic patients with paroxysmal nocturnal hemoglobinuria. AB - Platelet kinetic studies were performed in eight patients with paroxysmal nocturnal hemoglobinuria (PNH) but without symptoms or signs related to thrombosis. Indium-111 tropolonate labeled autologous platelets were used to determine mean platelet life and platelet production rate; total body images were also acquired. Three of seven patients with a thrombocytopenia showed a shortened mean platelet life, and the other five demonstrated a (nearly) normal platelet survival. Four of the seven patients with thrombocytopenia showed a decreased platelet production rate, but bone marrow cellularity was normo- to hypercellular. These observations suggest ineffective thrombopoiesis in 57% of the patients with PNH and thrombocytopenia. Total body imaging was performed during the platelet kinetic study in order to study organ uptake. Imaging of the abdominal vessels was demonstrated in all patients, suggesting enhanced adherence of platelets to the endothelium in patients with PNH. Since thrombosis of especially the abdominal vessels is a major cause of morbidity and mortality, the use of antithrombotic drugs might be considered despite the absence of abdominal complaints. PMID- 11732868 TI - Activated peripheral blood and endothelial cells in thalassemia patients. AB - BACKGROUND AND OBJECTIVES: Thalassemia patients have alterations in the expression of some activation and adhesion molecules on peripheral blood lymphocytes. We studied cell surface antigens on peripheral blood cells associated with the activation of these cells and soluble molecules produced by activated endothelium. DESIGN AND METHODS: We investigated the expression of CD11b, CD18, CD35, CD43, CD44, and CD69 on the peripheral blood monocytes, Cd11b, CD18, CD35, CD43, CD44, CD67 on peripheral blood neutrophils and CD38 and CD69 on peripheral blood lymphocytes. We studied 68 transfusion-dependent thalassemics (group A), 10 transfusion non-dependent thalassemics (group B), 18 beta thalassemia carriers (group C), and 28 normal individuals. Relative fluorescence intensity was used to determine the antigen density. Analysis was performed with an EPICS ELITE flow cytometer. Furthermore, soluble intercelullar adhesion molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), and E selectin, tumor necrosis factor (TNF) alpha, and interleukin (IL) 1beta were measured in the plasma of patients by enzyme-linked immunometric assay. RESULTS: The expression of CD11b, CD18, and CD69 on the monocytes of group A was significantly greater than in groups B and C and in controls, while CD44 was significantly downregulated in group A. CD11b, CD18, CD35, CD44, and CD67 on the surface of neutrophils and CD38 and CD69 on the surface of lymphocytes were also overexpressed in group A. CD44 was downregulated on the monocytes and upregulated on the neutrophils of the patients compared to controls. The levels of sICAM-1, sVCAM-1, E-selectin, TNF-alpha, and IL-1beta in the serum of patients in groups A and B were higher than those in group C and the controls. CONCLUSION: Endothelial activation markers are significantly increased in thalassemia patients, and activated blood cells circulate in the peripheral blood. These may be related to the vascular complications in these patients and might be useful markers for the follow-up of the vascular disease. PMID- 11732869 TI - Autologous transplantation in acute myeloid leukemia: peripheral blood stem cell harvest after mobilization in steady state by granulocyte colony-stimulating factor alone. AB - In order to determine whether granulocyte colony-stimulating factor (G-CSF) alone initiated during steady state was able to mobilize peripheral blood stem cells (PBSC) in acute myeloid leukemia (AML) and to assess predictive factors for engraftment after autologous PBSC transplantation, we studied 49 successive adult AML patients for whom autologous transplantation was planned between July 1994 and November 1998. G-CSF was used as priming agent and was initiated at least 4 weeks after the last day of chemotherapy, while neutrophil count was >0.5 x 10(9)/l and platelet count was >30 x 10(9)/l. A median of three aphereses was performed resulting in a median collection of 14.8 x 10(8) nucleated cells/kg containing 7.7 x 10(8) mononuclear cells/kg, 47.1 x 10(4) CFU-GM/kg, and 3.8 x 10(6) CD34+ cells/kg. A significant correlation was observed between nucleated cell, mononuclear cell, and CFU-GM yields, while no correlation was found with CD34+ cell yield. Recruitment was not significantly different in patients with CD34+ leukemic cells at the time of initial diagnosis when compared to that of those presenting with CD34- blastic cells. Thirty-three patients actually underwent transplantation. Reasons for not autografting were inadequate stem cell harvest (ten patients), early relapse (two patients), prolonged neutropenia (one patient), organ failure (two patients), or patient refusal (one patient). Median time to achieve a neutrophil count greater than 0.5 x 10(9)/l and platelet count >50 x 10(9)/l untransfused was 13 and 36 days, respectively. A predictive factor for a shorter period neutropenia and a shorter thrombopenia was a higher count of harvested nucleated cells (p < 0.01 and p = 0.02, respectively). A higher count of harvested cells was also a predictive factor for less red cell and platelet transfusions (p=0.03 and p=0.02, respectively). The number of CD34+ harvested PBSC was not predictive for engraftment. We conclude that PBSC mobilization with G-CSF alone initiated in steady state is a feasible, safe, and suitable procedure for harvesting cells in sight of autologous transplantation in adult acute myeloid leukemia. PMID- 11732870 TI - Predictive factors for a successful mobilization of peripheral blood CD34+ cells in multiple myeloma. AB - We analyzed the prognostic factors for a successful mobilization and peripheral blood stem cell collection in a series of 57 consecutive patients with multiple myeloma (MM); a new scoring system to predict an adequate mobilization in this subset of patients was also constructed. A total of 221 aphereses were performed in 57 patients with MM. The median time from diagnosis to mobilization was 12 months (range 4-120). Only one line of chemotherapy was administered before mobilization to 36 patients and two or more to 21. The median number of alkylating chemotherapy cycles was 6 (2-33). Two patients were mobilized in complete remission, 32 in partial response, and 23 in stable/progressive disease. Significant adverse prognostic factors for collecting 2.5 x 10(6) CD34+cells/kg or more were: a period of at least 12 months from diagnosis, at least six cycles of alkylating agents, and a plasma cell infiltration of 20% or more prior to mobilization. Patients with three risk factors had a probability of only 0.38 (95% CI 0.3-0.9) for adequate mobilization. Ten patients failed to mobilize; a period from diagnosis of 12 months or more and female sex were unfavorable factors. Patients with two risk factors had a probability of 0.50 (95% CI 0.2 0.8) for failing the mobilization procedure. These findings indicate that MM patients must be mobilized early in the course of the disease, with minimal disease burden before severe hematopoietic progenitor cell injury due to cumulative therapy. PMID- 11732871 TI - Relationship between bone marrow angiogenesis and plasma cell infiltration and serum beta2-microglobulin levels in patients with multiple myeloma. AB - There is growing evidence that angiogenesis is important not only in solid tumors but also in hematological malignancies. Recently, we found that bone marrow angiogenesis is a prognostic factor for disease-related survival in patients with multiple myeloma. In this report, we addressed the question of whether the microvessel density in bone marrow biopsies is correlated to other myeloma parameters, e.g., serum beta2-microglobulin (beta2-MG) and plasma cell infiltration in the bone marrow. In 22 multiple myeloma patients, immunohistochemical, CD34-stained, paraffin-embedded bone marrow biopsies before and after chemotherapy were studied. Microvessels were counted in 400x magnification, and the mean number of vessels per area in each sample was noted as the microvessel density (MVD). Pretreatment bone marrow MVD (median: 44, range: 11-175 vessels/mm2) correlated significantly with the bone marrow plasma cell infiltration (median: 30%, range: 5-90%, r = 0.642, P=0.001) and beta2-MG (median: 2.74, range: 1.4-26.1 mg/l, r = 0.749, P < 0.0005). In contrast, there was no correlation between posttreatment MVD and plasma cell infiltration or beta2-MG (median: MVD 31, range: 0-221 vessels/mm2, median plasma cell infiltration: 15%, range: 5-80%, r = 0.229, P = 0.306 and median beta2-MG: 2.65, range: 1-27.6 mg/l, r = -0.042, P = 0.853). These findings show that the strong correlations between bone marrow MVD and plasma cell infiltration as well as serum beta2-MG levels disappear after chemotherapy. The underlying mechanisms need further investigations. PMID- 11732872 TI - Full-dose CHOP chemotherapy combined with granulocyte colony-stimulating factor for aggressive non-Hodgkin's lymphoma in elderly patients: a prospective study. AB - We assessed the efficacy and safety of full-dose CHOP regimen plus granulocyte colony-stimulating factor to treat aggressive non-Hodgkin's lymphoma in elderly patients. Forty-two patients with untreated disease were included in this study, aged 70-79 years, with stage II or higher disease and a performance status of 0 3, without severe organ dysfunction. Of the 40 patients who could be evaluated 87.5% achieved complete remission, with a 4-year survival rate of 69% and a 3 year progression-free survival rate of 49%. When stratified by the International prognostic Index, the 4-year survival rate was 90.9% for the low and low intermediate risk group and 41.3% for the high-intermediate and high risk group, whereas the 3-year progression survival rate was 87.7% and 11.3%, respectively. Grade 3 or 4 hematological toxicity was found in 31 instances of granulocytopenia (77.5%) and 7 of anemia (17.5%). Nonhematological toxicity of grade 3 or 4 included pneumonia in two patients, heart failure in one, and gastrointestinal bleeding in one. Full-dose CHOP regimen with granulocyte colony-stimulating factor support could achieve a high-dose intensity in elderly patients whose general physical condition was good and hence achieved a high complete remission rate, but the disease often recurred within 2 years. Consequently, a new therapeutic strategy needs to be established, particularly for patients with high intermediate or high risk. PMID- 11732873 TI - Primary natural killer cell lymphoma of the lacrimal sac. AB - Primary lymphoid tumors of the lacrimal sac are quite rare, and all reported cases are of B-cell tumors with good prognosis. To our knowledge, this is the first case of primary natural killer (NK) cell lymphoma of the lacrimal sac. A 55 year-old woman presented with a lacrimal sac tumor, and histological diagnosis of NK cell lymphoma was made. Although disease was initially localized to the right lacrimal sac, it invaded into the adjacent ethmoidal sinus before chemotherapy was initiated (clinical stage IIE). Epstein-Barr virus (EBV)-encoded small RNA (EBER) was detected in lymphoma cells by in situ hybridization. Systemic chemotherapy combined with intrathecal chemotherapy followed by local radiotherapy was performed, and the patient achieved complete remission. However, shortly after completion of chemoradiotherapy, the lymphoma relapsed with rapid systemic dissemination. The disease was refractory to chemotherapy, and the patient eventually succumbed due to sepsis. PMID- 11732874 TI - Small bowel obstruction in acute myelogenous leukemia: stenosis or paralysis? AB - We describe a patient with acute myelogenous leukemia who suffered a small bowel obstruction on the second day of chemotherapy. The patient had to be operated immediately, and the terminal ileum and a part of the colon was removed. The resected specimen showed leukemic infiltration (chloroma) of the bowel with marked atrophy of the muscular layer. However, there was no complete stenosis. For this reason we believe that the reason for the acute abdominal symptoms on the second day of chemotherapy could be paralysis of the bowel due to muscular atrophy. PMID- 11732875 TI - Ogilvie's syndrome in acute myeloid leukemia: pharmacological approach with neostigmine. AB - Acute colonic pseudo-obstruction, the so-called Ogilvie's syndrome, is a rare and life-threatening digestive complication usually observed in critically ill patients. It is characterized by signs of large-bowel obstruction, without a mechanical cause, and has been reported in various settings, including acute leukemias as a complication of neutropenic enterocolitis after intensive chemotherapy. We describe the case of a young woman who, during the neutropenic phase following autologous bone marrow transplantation for relapsed acute myeloid leukemia, developed neutropenic enterocolitis complicated by an acute pseudo obstruction of descendent colon and sigma. This process was associated with sepsis and resolved with conservative therapy of the underlying condition, using granulocyte colony-stimulating factor and intravenous neostigmine. We discuss the management of this rare syndrome. PMID- 11732876 TI - Cytomegalovirus enteritis after autologous peripheral blood stem cell transplantation. AB - A 61-year-old male with non-Hodgkin's lymphoma (peripheral T-cell lymphoma, unspecified, clinical stage IVb) received autologous peripheral blood stem cell transplantation (PBSCT) during first remission. He was seropositive for cytomegalovirus (CMV) prior to autologous PBSCT. His posttransplant clinical course was complicated by refractory CMV enteritis, which manifested persistent abdominal pain, diarrhea, and bloody stool. Generally, gastrointestinal CMV disease is relatively rare after autologous PBSCT. However, our case indicates that CMV infection must be considered as a differential diagnosis in cases of unexplained hemorrhagic enteritis following autologous PBSCT. PMID- 11732877 TI - Sea-blue histiocytosis secondary to Niemann-Pick disease type B: a case report. AB - Sea-blue histiocytosis is a morphological finding that can be associated both with acquired conditions of increased cellular turnover and inborn errors of lipid metabolism. We report a rare case of sea-blue histiocytosis associated with a mild phenotype of Niemann-Pick disease (NPD) type B in a 44-year-old man who presented with splenomegaly and mild thrombocytopenia. Diagnosis was guided by the morphological finding in bone marrow smears of foamy and sea-blue histiocytes and confirmed by the measurement of acid lysosomal sphingomyelinase activity below normal values. NPD type B is a rare inborn error of metabolism, with a benign course and prognosis, while types A and C are always associated with severe neurological involvement. In our patient diagnosis was confirmed by the specific enzyme assay of leukocytes (deficiency in sphingomyelinase activity). This is a simple and noninvasive method that is useful whenever clinical and morphological finding are relevant, and a primary hematological disorder has been ruled out. PMID- 11732878 TI - Pregnancy induced hemolytic anemia: an unexplained entity. AB - We present the case of a young primigravida who developed severe life threatening hemolytic anemia in the last trimester of three successive pregnancies with spontaneous recovery after each delivery and remained normal during the entire nongravid state. Corticosteroid and high-dose intravenous immunoglobulin therapy, although reported as useful, was ineffective in our case. She was managed only with the support with top-up blood transfusions. Extensive investigations were carried out to determine the cause of hemolysis, but these proved unfruitful. Fewer than two scores of such cases have been described in the literature. The paucity of such cases merits its presentation. It is suggested that this anemia should be referred to as "pregnancy-induced hemolytic anemia." PMID- 11732879 TI - Ocular tear film bubble counts after recreational compressed air diving. AB - Other authors have demonstrated an increase in tear film bubble counts following dry, compressed air dives. We examined the lower tear film meniscus for the presence of bubbles in 42 divers after compressed air dives on a single day and in 11 divers undergoing repetitive, multi-day diving exposures over 5 days. After diving, bubble counts increased significantly (P < 0.01) from predive values. From a predive median (inter-quartile range) of 0 (0-0.33) bubbles/eye, single day divers reached a maximum bubble count at 48 h after diving of 1 (0-2.25) bubbles/eye. Similarly, from a predive count of 0.33 (0-1) bubbles/eye, multi-day divers had increased bubble counts from 24 h following their first dive until 24 h following their final dive when counts were 1.67 (0.92-3.08) bubbles/eye. Bubble counts were not significantly correlated with inert gas load, body mass index, age, or diving experience. We confirm that tear film bubble counts are raised after wet compressed air diving as previously described following dry diving. PMID- 11732880 TI - Increased lung compliance in response to a moderate hyperoxic exposure. AB - With deep saturation diving a reduction in vital capacity caused by oxygen toxicity may be opposed by a training effect of respiratory muscles due to increased gas density and work of breathing. We measured lung and chest wall mechanics before and after a 28-day saturation dive to a pressure of 0.25 MPa with the same profile of oxygen exposure as in a deep dive to a pressure of 3.7 MPa; 40 kPa during the isopression phase and 50 kPa during the decompression phase. Eight males aged 22-28 yr served as subjects. The measurements included dynamic lung volumes, static lung compliance, lung recoil pressure, and maximal respiratory pressures. Only one subject had decreased lung compliance and increased recoil pressure after the dive. The others had an increase in compliance and decrease in recoil pressure. There was a significant increase in inspiratory lung compliance (P = 0.041) and a trend for a decrease in lung recoil pressure (P = 0.061). We found no change in forced vital capacity, but decreases in forced expired volume in 1 s (P = 0.049) and forced midexpiratory flow rate (P = 0.009) were noted. There were no changes in maximal respiratory pressures. These findings are opposite to the classical findings associated with pulmonary oxygen toxicity. The results may reflect an increase in surfactant production and turnover as an early adaptive response to hyperoxic stress. PMID- 11732881 TI - Personality characteristics of South African navy divers. AB - The first study aimed to determine the extent to which the 16 Personality Factor Questionnaire (16PF) can be used to describe successful divers in the South African Navy. The 16PF profiles of 100 divers were analyzed to describe the personality characteristics of naval divers. Results indicate that four personality factors were most descriptive of the sample. They were Group orientation, Enthusiasm, Adventurousness, and Confidence. These factors seemed appropriate when discussed from an environmental demand perspective. The second study aimed to determine the extent to which the 16 PF can be used to differentiate between divers and submariners in the South African Navy. The two groups differed significantly on two factors--divers scored higher on Enthusiasm and submariners scored higher on Conservatism. PMID- 11732882 TI - Orbital hemorrhage following face-mask barotrauma. AB - A 41-yr-old female recreational diver suffered an orbital hemorrhage following face-mask barotrauma This case illustrates the potential for significant orbital injury to occur as a result of face-mask barotrauma Barotraumatic orbital hemorrhage is an unusual disorder, but recognition of this entity is important for diving physicians. The signs and symptoms should not be mistaken for decompression sickness or arterial gas embolism. Patients with barotraumatic orbital hemorrhage require emergency referral to an ophthalmologist so that potentially vision-threatening ocular complications may be identified and treated. Barotrauma may result in several different types of cranial neuropathies and should be recognized as another possible cause for neurologic deficits following a dive. PMID- 11732883 TI - Management of ischemic hemorrhagic cystitis with hyperbaric oxygen therapy. AB - We report a case of a T8 paraplegic woman who experienced necrosis of the bladder due to inadvertent chronic over distention. After bladder repair, the patient suffered from severe ischemic hemorrhagic cystitis. Despite many attempts to control the bleeding with standard treatments, the hematuria continued. Although hyperbaric oxygen (HBO2) therapy has not been proven to be helpful in patients with hemorrhagic cystitis not related to radiation or chemotherapy, the physiopathologic mechanisms of the cysttis, involving ischemia, led us to believe that outpatient would benefit from HBO2 therapy in analogy to the approved use of HBO2 therapy in radiation cystitis. Since the use of HBO2 therapy has not been previously reported in this situation, we met with some hesitation from the HBO2 therapy department After reviewing the literature and discussing the case, a medical panel reached a consensus to allow us to commence the treatment The patient had a remarkable response, and remains without hematuria 6 mo. after treatment. PMID- 11732884 TI - Recreational scuba divers' aversion to low-frequency underwater sound. AB - Increasing use of active low-frequency sonar by submarines and ships raises the risk of accidental exposure of recreational divers to low-frequency underwater sound (LFS). This study aimed to characterize the subjective responses of recreational scuba divers to LFS to ascertain the extent to which LFS may impact their enjoyment, comfort, or time spent underwater. Seventeen male and nine female recreational scuba divers participated. Diving was conducted in an acoustically transparent tank located within a larger anechoic pool. Subjects wore scuba gear and were positioned I m below the surface in a prone position. The sound transducer was located 4 m directly below the diver's head. Sound exposures consisted of three signal types (pure tone, 30 Hz hyperbolic sweep up, and 30 Hz hyperbolic sweep down) each presented at six center frequencies from 100 to 500 Hz and six sound pressure levels(SPL) ranging from 130 to 157 dB re 1 microPa. The duration of each sound exposure was 7 s. Subjects responded via an underwater console to rate aversion to LFS on a category-ratio scale, and to indicate the presence or absence of vibration of any body part. Aversion to LFS and the percent incidence of vibration increased as the SPL increased. The percent incidence of vibration decreased linearly with increasing frequency. At the highest SPL the probability that an aversion rating would exceed Very Severe (7 on the category-ratio scale) was predicted to be 19%. There was no significant difference in aversion among signal types. The 100 Hz frequency was the most aversive frequency (P < 0.05). A plot of aversion vs. frequency showed a U-shaped function with minimum aversion at 250 Hz. In conclusion, diver aversion to LFS is dependent upon SPL and center frequency. The highest aversion rating was given for 100 Hz, this frequency corresponded with the greatest probability of detecting vibration. Factors other than vibration seem to account for aversion to the highest frequencies. Our data suggest that LFS exposures up to 145 dB re 1 microPa at frequencies between 100 and 500 Hz will have minimal impact on the recreational diver. PMID- 11732885 TI - Gastrointestinal tract as a part of immune defence. AB - The paper is devoted to reviewing the function of gastrointestinal mucosal immune system. Gut-associated lymphoid tissue (GALT) provides the host with protective mechanisms against invasion by potential pathogens across the mucosal surface, and on the other hand, it plays a role in the development of induced tolerance against harmless products of digestion and the normal intestinal flora, all of which are potentially immunogenic. First the organization of GALT, its afferent and efferent limbs are described, then the normal B cell homeostasis and finally the induced oral tolerance are discussed. Knowledge of mechanisms of mucosal immune response is essential to complete understanding of the pathogenesis and treatment of inflammatory gastrointestinal diseases as well. PMID- 11732886 TI - Gastrointestinal immunology: cell types in the lamina propria--a morphological review. AB - Defense mechanisms--including immune responses--of the gastrointestinal (GI) system rely on a delicate balance of multidirectional interactions of different components of the GI mucosa. The majority of the cells involved in immune reactions are in the lamina propria (LP) and in the submucosa. Several biologically active substances (enzymes, neurotransmitters, humoral mediators) and their receptors have been reported to be present in LP cells. These cells are in close morphological connections with the surface epithelial cells and with the surrounding vessels and nerve fibers, suggesting a functional association with them. In this paper cell types of the LP will be reviewed from a morphological aspect. In summary, LP cells can be classified as basic structural elements (fibroblasts, fibrocytes, vascular endothelial and smooth muscle cells), blood cells (granulocytes, mast cells, macrophages, T and B lymphocytes, plasma cells), and occasional epithelial and endocrine cells of the surface epithelium. Nerve fibers and terminals, but not neuronal perikarya, can also be seen in the LP. The appearance and the proportion of LP cells strongly depend on the functional activity of the GI system at any given time. Their number and distribution might be significantly altered in certain pathological conditions (infections, inflammations, ulcerations, and other GI disorders). We hope, this review may help clinicians, pathologists, and researchers in the recognition of LP cell types, and in demonstrating their activation, migration and proliferation in different physiological and pathological conditions. PMID- 11732887 TI - Immunological aspects of Helicobacter pylori infection. AB - Host defence against Helicobacter pylori infection is a complex system of both specific and non-specific reactions. Among the non-specific defense mechanisms acting on bacteria before they reach the mucus layer in the stomach are digestive enzymes, lyzozyme, lactoferrin and other components with antimicrobal activity. The mucus layer is the final non-specific barrier against the bacteria reaching the gastric mucosa cells. On reaching the gastric mucosa Helicobacter pylori adheres to epithelial cells and bacterial antigens, chemotaxins and other components are liberated. Helicobacter pylori antigens are presented to immunate B lymphocytes, which interact with T-helper lymphocytes to become mature IgA-, IgD-, IgE-, IgG and Ig-M producing plasma cells. IgA dimers of secretory IgA are secreted through the gastric epithelium. IgE antibodies bind to basophils, which mature to histamine-producing mast cells. Histamine activates the acid production in the stomach and contributes to the chronic inflammation and tissue destruction. In addition, T lymphocytes are possible activated by Helicobacter pylori and contribute to the chronic inflammation. PMID- 11732888 TI - Genetic associations and immunopathogenesis of coeliac disease. AB - Coeliac disease is the most common disorder with malabsorption of the small instestine, caused by the gluten fraction of cereals in genetically predisposed individuals. Gluten peptides are efficiently presented by coeliac disease specific HLA-DQ2- and HLA-DQ8-positive antigen-presenting cells, and thus drive the antigen-presenting cells, predominantly in the connective tissue of the lamina propria. The studying of the recently explored autoantibodies against tissue transglutaminase brought us further in the understanding of the pathophysiology of coeliac disease. The spreading of reliable serologic methods modified our knowledge on the clinical picture and prevalence of the disease. Long-standing untreated coeliac disease, even if clinically silent, predisposes for other autoimmune diseases. Therefore, population screening for immunoglobulin A antibodies to tissue transglutaminase seems justified. PMID- 11732889 TI - Immunology of inflammatory bowel diseases. AB - The pathogenesis of inflammatory diseases involves complex interactions among genetic, environmental and immunologic factors, where the immune system plays a crucial role. The initiating factor may be a commonly present environmental antigen, but the defective mucosal immune response can't downregulate the inflammatory process, the resulting in chronic inflammation and tissue damage. The abnormal amplification of the immune system seems more likely to be a secondary and not a primary factor in the disease. IBD patients exhibit abnormalities in epithelial antigen presentation, and in the cellular and humoral immune system. The role of autoimmunity is unclear in IBD. The inflammatory mediator production and balance is also altered in IBD. Crohn's disease is characterised by a Th1 cytokine profile, while ulcerative colitis is characterised rather by a Th2 cytokine profile. Recent advances in the understanding of the pathogenesis of IBD led to major changes in the treatment of the disease. The newer agents target the key mediators involved in the inflammatory process. PMID- 11732890 TI - Pathogenesis and treatment of autoimmune hepatitis and chronic viral hepatitis B and C. AB - Immune mechanisms play a role in autoimmune hepatitis which is considered as "idiopathic" inflammatory liver disease of unknown etiology. However, even chronic viral hepatitis B and C have also features suggesting the importance of immunopathogenesis in their development. This paper discusses the major genetical and immunological factors in the above-mentioned chronic liver diseases and briefly summarizes their therapeutic modalities. PMID- 11732891 TI - An efficient high-throughput resonance assignment procedure for structural genomics and protein folding research by NMR. AB - Sequence specific resonance assignment is the primary requirement for all investigations of proteins by NMR methods. In the present postgenomic era where structural genomics and protein folding have occupied the center stage of NMR research, there is a high demand on the speed of resonance assignment, whereas the presently available methods based either on NOESY or on some triple-resonance experiments are rather slow. They also have limited success with unfolded proteins because of the lack of NOEs, and poor dispersion of amide and carbon chemical shifts. This paper describes an efficient approach to rapid resonance assignment that is suitable for both folded and unfolded proteins, making use of the triple-resonance experiments described recently [HNN and HN(C)N]. It has three underlying principles. First, the experiments exploit the (15)N chemical shift dispersions which are generally very good for both folded and unfolded proteins, along two of the three dimensions; second, they directly display sequential amide and (15)N correlations along the polypeptide chain, and third, the sign patterns of the diagonal and the sequential peaks originating from any residue are dependent on the nature of the adjacent residues, especially the glycines and the prolines. These lead to so-called "triplet fixed points" which serve as starting points and/or check points during the course of sequential walks, and explicit side chains assignment becomes less crucial for unambiguous backbone assignment. These features significantly enhance the speed of data analysis, reduce the amount of experimentation required, and thus result in a substantially faster and unambiguous assignment. Following the amide and (15)N assignments, the other proton and carbon assignments can be obtained in a straightforward manner, from the well-established three-dimensional triple resonance experiments. We have successfully tested the new approach with different proteins in the molecular mass range of 10-22 kDa, and for illustration, we present here the backbone results on the HIV-1 protease-tethered dimer (molecular mass approximately 22 kDa), both in the folded and in the unfolded forms, the two ends of the folding funnel. We believe that the new assignment approach will be of great value for both structural genomics and protein folding research by NMR. PMID- 11732892 TI - Spontaneous in vitro formation of supramolecular beta-amyloid structures, "betaamy balls", by beta-amyloid 1-40 peptide. AB - The concentration of beta-amyloid peptide (Abeta), x-42 or x-40 amino acids long, increases in brain with the progression Alzheimer's disease (AD). These peptides are deposited extracellularly as highly insoluble fibrils that form densities of amyloid plaques. Abeta fibrillization is a complex polymerization process preceded by the formation of oligomeric and prefibrillar Abeta intermediates. In some of our in vitro studies, in which the kinetics of intermediate steps of fibril formation were examined, we used concentrations of synthetic Abeta that exceed what is normally employed in fibrillization studies, 300-600 microM. At these concentrations, in a cell free system and under physiological conditions, Abeta 1-40 peptide (Abeta40) forms fibrils that spontaneously assemble into clearly defined spheres, "betaamy balls", with diameters of approximately 20-200 microm. These supramolecular structures show weak birefringence with Congo red staining and high stability with prolonged incubation times (at least 2 weeks) at 30 degrees C, freezing, and dilution in H(2)O. At 600 microM, they are detected after incubation for approximately 20 h. Abeta peptide 1-42 (Abeta42) lacks the ability to form betaamy balls but accelerates Abeta40 betaamy ball formation at low stoichiometric levels (1:20 Abeta42:Abeta40 ratio). Abeta42 levels above this (=10-50% w/w) impede Abeta40 betaamy ball formation. Using light (LM) and electron microscopy (EM), this study examines the gross morphology and ultrastructure of Abeta40 betaamy balls and their time course of formation, in the absence and presence of Abeta42, along with some stability measures. As spheres of a misfolded protein, betaamy balls resemble both AD Abeta senile plaques and neuronal inclusion bodies associated with other neurodegenerative diseases. PMID- 11732893 TI - Main chain and side chain dynamics of oxidized flavodoxin from Cyanobacterium anabaena. AB - Oxidized flavodoxin from Cyanobacterium anabaena PCC 7119 is used as a model system to investigate the fast internal dynamics of a flavin-bearing protein. Virtually complete backbone and side chain resonance NMR assignments of an oxidized flavodoxin point mutant (C55A) have been determined. Backbone and side chain dynamics in flavodoxin (C55A) were investigated using (15)N amide and deuterium methyl NMR relaxation methods. The squared generalized order parameters (S(NH)(2)) for backbone amide N-H bonds are found to be uniformly high ( approximately 0.923 over 109 residues in regular secondary structure), indicating considerable restriction of motion in the backbone of the protein. In contrast, methyl-bearing side chains are considerably heterogeneous in their amplitude of motion, as indicated by obtained symmetry axis squared generalized order parameters (S(axis)(2)). However, in comparison to nonprosthetic group-bearing proteins studied with these NMR relaxation methods, the side chains of oxidized flavodoxin are unusually rigid. PMID- 11732894 TI - Site-directed tryptophan fluorescence reveals the solution structure of tear lipocalin: evidence for features that confer promiscuity in ligand binding. AB - The solution structure of human TL was deduced from the position of the emission peaks after site-directed tryptophan fluorescence (SDTF). The fluorescent amino acid tryptophan was sequentially substituted for each native amino acid in the sequence. Characteristic periodicities for eight beta-strands that comprise the beta-barrel and three alpha-helices were identified. The putative beta-strand I was relatively exposed to solvent, suggesting it does not participate in the formation of the beta-barrel. The beta-strands A and F contain beta-bulges. The average lambda(max) of emission maxima reveals that strand D is at the edge of the barrel and beta-strand H interacts with the main alpha-helical domain. On the basis of the SDTF data, a 3D homology model was constructed for TL and compared to the known crystallographic structures of RBP and beta-lactoglobulin. The small size and splayed open configuration of the E-F hairpin facilitate access of ligands into the cavity mouth of TL as compared to that of RBP with a long overhanging loop that restricts access. In the model of TL, four alanine residues are positioned in the binding site as compared to bulkier residues in the corresponding positions of beta-lactoglobulin. Substitution of A51, A66, A86 to Trp results in a 3-4-fold decrease in binding affinity. The data suggest that the smaller side chains of Ala provide more capacity in the cavity of TL than the bulkier side chains (I56, I71, V92) in the cavity of beta-lactoglobulin. The structural features provide an explanation for the promiscuous binding characteristics exhibited by TL. SDTF provides a general approach for determining the solution structure of many proteins and enhances homology modeling in the absence of high sequence identity. PMID- 11732895 TI - The structure of L-ribulose-5-phosphate 4-epimerase: an aldolase-like platform for epimerization. AB - The structure of L-ribulose-5-phosphate 4-epimerase from E. coli has been solved to 2.4 A resolution using X-ray diffraction data. The structure is homo tetrameric and displays C(4) symmetry. Each subunit has a single domain comprised of a central beta-sheet flanked on either side by layers of alpha-helices. The active site is identified by the position of the catalytic zinc residue and is located at the interface between two adjacent subunits. A remarkable feature of the structure is that it shows a very close resemblance to that of L-fuculose-1 phosphate aldolase. This is consistent with the notion that both enzymes belong to a superfamily of epimerases/aldolases that catalyze carbon-carbon bond cleavage reactions via a metal-stabilized enolate intermediate. Detailed inspection of the epimerase structure, however, indicates that despite the close overall structural similarity to class II aldolases, the enzyme has evolved distinct active site features that promote its particular chemistry. PMID- 11732896 TI - Catalysis and binding in L-ribulose-5-phosphate 4-epimerase: a comparison with L fuculose-1-phosphate aldolase. AB - L-Ribulose-5-phosphate (L-Ru5P) 4-epimerase and L-fuculose-1-phosphate (L-Fuc1P) aldolase are evolutionarily related enzymes that display 26% sequence identity and a very high degree of structural similarity. They both employ a divalent cation in the formation and stabilization of an enolate during catalysis, and both are able to deprotonate the C-4 hydroxyl group of a phosphoketose substrate. Despite these many similarities, subtle distinctions must be present which allow the enzymes to catalyze two seemingly different reactions and to accommodate substrates differing greatly in the position of the phosphate (C-5 vs C-1). Asp76 of the epimerase corresponds to the key catalytic acid/base residue Glu73 of the aldolase. The D76N mutant of the epimerase retained considerable activity, indicating it is not a key catalytic residue in this enzyme. In addition, the D76E mutant did not show enhanced levels of background aldolase activity. Mutations of residues in the putative phosphate-binding pocket of the epimerase (N28A and K42M) showed dramatically higher values of K(M) for L-Ru5P. This indicates that both enzymes utilize the same phosphate recognition pocket, and since the phosphates are positioned at opposite ends of the respective substrates, the two enzymes must bind their substrates in a reversed or "flipped" orientation. The epimerase mutant D120N displays a 3000-fold decrease in the value of k(cat), suggesting that Asp120' provides a key catalytic acid/base residue in this enzyme. Analysis of the D120N mutant by X-ray crystallography shows that its structure is indistinguishable from that of the wild-type enzyme and that the decrease in activity was not simply due to a structural perturbation of the active site. Previous work [Lee, L. V., Poyner, R. R., Vu, M. V., and Cleland, W. W. (2000) Biochemistry 39, 4821-4830] has indicated that Tyr229' likely provides the other catalytic acid/base residue. Both of these residues are supplied by an adjacent subunit. Modeling of L-Ru5P into the active site of the epimerase structure suggests that Tyr229' is responsible for deprotonating L-Ru5P and Asp120' is responsible for deprotonating its epimer, D-Xu5P. PMID- 11732897 TI - A structural view of the action of Escherichia coli (lacZ) beta-galactosidase. AB - The structures of a series of complexes designed to mimic intermediates along the reaction coordinate for beta-galactosidase are presented. These complexes clarify and enhance previous proposals regarding the catalytic mechanism. The nucleophile, Glu537, is seen to covalently bind to the galactosyl moiety. Of the two potential acids, Mg(2+) and Glu461, the latter is in better position to directly assist in leaving group departure, suggesting that the metal ion acts in a secondary role. A sodium ion plays a part in substrate binding by directly ligating the galactosyl 6-hydroxyl. The proposed reaction coordinate involves the movement of the galactosyl moiety deep into the active site pocket. For those ligands that do bind deeply there is an associated conformational change in which residues within loop 794-804 move up to 10 A closer to the site of binding. In some cases this can be inhibited by the binding of additional ligands. The resulting restricted access to the intermediate helps to explain why allolactose, the natural inducer for the lac operon, is the preferred product of transglycosylation. PMID- 11732898 TI - Restricting the ligand-linked heme movement in Scapharca dimeric hemoglobin reveals tight coupling between distal and proximal contributions to cooperativity. AB - Cooperative ligand binding in the dimeric hemoglobin from the blood clam Scapharca inaequivalvis results primarily from tertiary, rather than quaternary, structural changes. Ligand binding is coupled with conformational changes of key residues, including Phe 97, which is extruded from the proximal heme pocket, and the heme group, which moves deeper into the heme pocket. We have tested the role of the heme movement in cooperative function by mutating Ile 114, at the base of the heme pocket. Replacement of this residue with a Met did not disturb the hemoglobin structure or significantly alter equilibrium ligand binding properties. In contrast, substitution with a Phe at position 114 inhibits the ligand-linked movement of the heme group, and substantially reduces oxygen affinity and cooperativity. As the extent of heme movement to the normal position of the ligated state is diminished, Phe 97 is inhibited from its movement into the interface upon ligand binding. These results indicate a tight coupling between these two key cooperative transitions and suggest that the heme movement may be an obligatory trigger for expulsion of Phe 97 from the heme pocket. PMID- 11732899 TI - 9-Anthroylnitrile binding to serine-181 in myosin subfragment 1 as revealed by FRET spectroscopy and molecular modeling. AB - It has been shown that one of the 12 serine residues within the 23 kDa segment of myosin subfragment 1 can be covalently modified with a fluorescent probe 9 anthroylnitrile (ANN) [Hiratsuka, T. (1989) J. Biol. Chem. 264 (30), 18188 18194]. To identify the exact binding site of the probe, the distances between the bound ANN as donor and acceptors in known positions (Lys-553 or Cys-707) of the myosin head were determined by using fluorescence resonance energy transfer. Comparison of the spectroscopic results with distances obtained from the atomic model of subfragment 1 revealed that ANN binds to Ser-181. The result was in good agreement with the assumptions of Andreev and co-workers [Andreev, O. A., et al. (1995) J. Muscle Res. Cell Motil. 16 (4), 353-367]. This conclusion was further supported by protein modeling calculations. The results presented herein might bring ANN into the focus when the molecular mechanism and effects of the binding of ATP and its subsequent hydrolysis are studied. PMID- 11732900 TI - Steric hindrance as a basis for structure-based design of selective inhibitors of protein-tyrosine phosphatases. AB - Utilizing structure-based design, we have previously demonstrated that it is possible to obtain selective inhibitors of protein-tyrosine phosphatase 1B (PTP1B). A basic nitrogen was introduced into a general PTP inhibitor to form a salt bridge to Asp48 in PTP1B and simultaneously cause repulsion in PTPs containing an asparagine in the equivalent position [Iversen, L. F., et al. (2000) J. Biol. Chem. 275, 10300-10307]. Further, we have recently demonstrated that Gly259 in PTP1B forms the bottom of a gateway that allows easy access to the active site for a broad range of substrates, while bulky residues in the same position in other PTPs cause steric hindrance and reduced substrate recognition capacity [Peters, G. H., et al. (2000) J. Biol. Chem. 275, 18201-18209]. The current study was undertaken to investigate the feasibility of structure-based design, utilizing these differences in accessibility to the active site among various PTPs. We show that a general, low-molecular weight PTP inhibitor can be developed into a highly selective inhibitor for PTP1B and TC-PTP by introducing a substituent, which is designed to address the region around residues 258 and 259. Detailed enzyme kinetic analysis with a set of wild-type and mutant PTPs, X-ray protein crystallography, and molecular modeling studies confirmed that selectivity for PTP1B and TC-PTP was achieved due to steric hindrance imposed by bulky position 259 residues in other PTPs. PMID- 11732901 TI - Substrate deactivation of phenylalanine-sensitive 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase by erythrose 4-phosphate. AB - 3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAH7PS, EC 4.1.2.15) catalyzes the condensation of phosphoenolpyruvate (PEP) with erythrose 4 phosphate (E4P) to give DAH7P via an ordered sequential mechanism. In the absence of PEP (the first substrate to bind), E4P binds covalently to the phenylalanine sensitive DAH7PS of Escherichia coli, DAH7PS(Phe), deactivating the enzyme. Activity is restored on addition of excess PEP but not if deactivation was carried out in the presence of sodium cyanoborohydride. Electrospray mass spectrometry indicates that a single E4P is bound to the protein. These data are consistent with a slow, reversible Schiff base reaction of the aldehydic functionality of E4P with a buried lysine. Molecular modeling indicates that Lys186, a residue at the base of the substrate-binding cavity involved in hydrogen bonding with PEP, is well placed to react with E4P forming an imine linkage that is substantially protected from solvent water. PMID- 11732902 TI - Structure-guided programming of polyketide chain-length determination in chalcone synthase. AB - Chalcone synthase (CHS) belongs to the family of type III polyketide synthases (PKS) that catalyze formation of structurally diverse polyketides. CHS synthesizes a tetraketide by sequential condensation of three acetyl anions derived from malonyl-CoA decarboxylation to a p-coumaroyl moiety attached to an active site cysteine. Gly256 resides on the surface of the CHS active site that is in direct contact with the polyketide chain derived from malonyl-CoA. Thus, position 256 serves as an ideal target to probe the link between cavity volume and polyketide chain-length determination in type III PKS. Functional examination of CHS G256A, G256V, G256L, and G256F mutants reveals altered product profiles from that of wild-type CHS. With p-coumaroyl-CoA as a starter molecule, the G256A and G256V mutants produce notably more tetraketide lactone. Further restrictions in cavity volume such as that seen in the G256L and G256F mutants yield increasing levels of the styrylpyrone bis-noryangonin from a triketide intermediate. X-ray crystallographic structures of the CHS G256A, G256V, G256L, and G256F mutants establish that these substitutions reduce the size of the active site cavity without significant alterations in the conformations of the polypeptide backbones. The side chain volume of position 256 influences both the number of condensation reactions during polyketide chain extension and the conformation of the triketide and tetraketide intermediates during the cyclization reaction. These results viewed in conjunction with the natural sequence variation of residue 256 suggest that rapid diversification of product specificity without concomitant loss of substantial catalytic activity in related CHS-like enzymes can occur by site-specific evolution of side chain volume at position 256. PMID- 11732903 TI - Loss of serotonin oxidation as a component of the altered substrate specificity in the Y444F mutant of recombinant human liver MAO A. AB - To investigate the roles of tyrosyl residues located near the covalent 8alpha-S cysteinyl FAD in monoamine oxidase A (MAO A) and to test the suggestion that MAO A and plant polyamine oxidase may have structural homology, tyrosyl to phenylalanyl mutants of MAO A at positions 377, 402, 407, 410, 419, and 444 were constructed and expressed in Saccharomyces cerevisiae. All mutant enzymes were expressed and exhibited lower specific activities as compared to WT MAO A using kynuramine as substrate. The lowest specific activities in this assay are exhibited by the Y407F and Y444F mutant enzymes. On purification and further characterization, these two mutants were found to each contain covalent FAD. Both mutant enzymes are irreversibly inhibited by the MAO A inhibitor clorgyline and exhibit binding stoichiometries of 0.54 (Y407F) and 0.95 (Y444F) as compared to 1.05 for WT MAO A. Y444F MAO A oxidizes kynuramine with a k(cat) <2% of WT enzyme and is greater than 100-fold slower in catalyzing the oxidation of phenylethylamine or of serotonin. In contrast, Y444F MAO A oxidizes p-CF(3) benzylamine at a rate 25% that of WT enzyme. Steady state and reductive half reaction stopped-flow data using a series of para-substituted benzylamine analogues show Y444F MAO A exhibits quantitative structure activity relationships (QSAR) properties on analogue binding and rates of substrate oxidation very similar to that exhibited by the WT enzyme (Miller and Edmondson (1999) Biochemistry 38, 13670): log K(d) = -(0.37 +/- ()()0.07)V(W)(x0.1) - 4.5 +/- 0.1; log k(red) = +(2.43 +/- 0.19)sigma + 0.17 +/- 0.05. The Y444F MAO A mutant also exhibits similar QSAR properties on the binding of phenylalkyl side chain amine analogues as WT enzyme: log K(i) = (4.37 +/- 0.51)E(S) + 1.21 +/- 0.77. These data show that mutation of Y444F in MAO A results in a mutant that has lost its ability to efficiently oxidize serotonin (its physiological substrate) but, however, exhibits unaltered quantitative structure-activity parameters in the binding and rate of benzylamine analogues. The mechanism of C-H abstraction is therefore unaltered. The suggestion that polyamine oxidase and monoamine oxidase may have structural homology appears to be valid as regards Y444 in MAO A and Y439 in plant polyamine oxidase. PMID- 11732905 TI - In vitro reconstitution and analysis of the chain initiating enzymes of the R1128 polyketide synthase. AB - Biosynthesis of the carbon chain backbone of the R1128 substances is believed to involve the activity of a ketosynthase/chain length factor (ZhuB/ZhuA), an additional ketosynthase (ZhuH), an acyl transferase (ZhuC), and two acyl carrier proteins (ACPs; ZhuG and ZhuN). A subset of these proteins initiate chain synthesis via decarboxylative condensation between an acetyl-, propionyl-, isobutyryl-, or butyryl-CoA derived primer unit and a malonyl-CoA derived extender unit to yield an acetoacetyl-, beta-ketopentanoyl-, 3-oxo-4 methylpentanoyl-, or beta-ketohexanoyl-ACP product, respectively. To investigate the precise roles of ZhuH, ZhuC, ZhuG, and ZhuN, each protein was expressed in Escherichia coli and purified to homogeneity. Although earlier reports had proposed that ZhuC and its homologues played a role in primer unit selection, direct in vitro analysis of ZhuC showed that it was in fact a malonyl-CoA:ACP malonyltransferase (MAT). The enzyme could catalyze malonyl transfer but not acetyl- or propionyl-transfer onto R1128 ACPs or onto ACPs from other biosynthetic pathways, suggesting that ZhuC has broad substrate specificity with respect to the holo-ACP substrate but is specific for malonyl-CoA. Thus, ZhuC supplies extender units to both the initiating and elongating ketosynthases from this pathway. To interrogate the primer unit specificity of ZhuH, the kinetics of beta-ketoacyl-ACP formation in the presence of various acyl-CoAs and malonyl-ZhuG were measured. Propionyl-CoA and isobutyryl-CoA were the two most preferred substrates of ZhuH, although acetyl-CoA and butyryl-CoA could also be accepted and elongated. This specificity is not only consistent with earlier reports demonstrating that R1128B and R1128C are the major products of the R1128 pathway in vivo, but is also in good agreement with the properties of the ZhuH substrate binding pocket, as deduced from a recently solved crystal structure of the enzyme. Finally, to investigate the molecular logic for the occurrence of not one but two ACP genes within the R1128 gene cluster, the inhibition of ZhuH-catalyzed formation of beta-ketopentanoyl-ACP was quantified in the presence of apo-ZhuG or apo-ZhuN. Both apo-proteins were comparable inhibitors of the ZhuH catalyzed reaction, suggesting that the corresponding apo-proteins can be used interchangeably during chain initiation. Together, these results provide direct biochemical insights into the mechanism of chain initiation of an unusual bacterial aromatic PKS. PMID- 11732904 TI - Geranylgeranylglyceryl phosphate synthase. Characterization of the recombinant enzyme from Methanobacterium thermoautotrophicum. AB - Geranylgeranylglyceryl diphosphate synthase (GGGP synthase) catalyzes alkylation of (S)-glyceryl phosphate [(S)-GP] by geranylgeranyl diphosphate (GGPP) to produce (S)-geranylgeranylglyceryl phosphate [(S)-GGGP]. This reaction is the first committed step in the biosynthesis of ether-linked membrane lipids in Archaea. The gene encoding GGGP synthase from Methanobacterium thermoautotrophicum was cloned using probes designed from the N-terminal sequence determined from the purified enzyme. The open reading frame, which encoded a protein of 245 amino acids, was inserted into a pET expression vector and expressed in Escherichia coli. The recombinant GGGP synthase was purified to homogeneity. The enzyme is active as a homopentamer, as determined by size exclusion chromatography and equilibrium sedimentation experiments. GGGP synthase has optimal activity at 55 degrees C in pH 8.0 buffer containing 1 mM MgCl(2). V(max) = 4.0 +/- 0.1 micromol min(-1) mg(-1) (k(cat) = 0.34 +/- 0.03 s(-1) for pentameric GGGP synthase assuming all subunits are fully active), K(m)((S)-GP) = 13.5 +/- 1.0 microM, and K(m)(GGPP) = 506 +/- 47 nM. These steady-state catalytic constants were identical to those for enzyme isolated from cell extracts of M. thermoautotrophicum [Chen, A., Zhang, D., and Poulter, C. D. (1993) J. Biol. Chem. 268, 21701-21705]. Alignment of seven putative archaeal GGGP synthase sequences revealed a number of highly conserved residues consisting of five aspartate/glutamates, three serine/threonines, two prolines, and five glycines, including a conserved GGG motif. PMID- 11732906 TI - Inhibition of tyrosine phenol-lyase from Citrobacter freundii by 2-azatyrosine and 3-azatyrosine. AB - The interactions of 2-azatyrosine and 3-azatyrosine with tyrosine phenol-lyase (TPL) from Citrobacter freundii have been examined. 2-Aza-DL-tyrosine and 3-aza DL-tyrosine were synthesized by standard methods of amino acid synthesis, while the L-isomers were prepared from 3-hydroxypyridine and 2-hydroxypyridine, respectively, with TPL (Watkins, E. B., and Phillips, R. S. (2001) Bioorg. Med. Chem. Lett. 11, 2099-2100). 3-Azatyrosine was examined as a potential transition state analogue inhibitor of TPL. Both compounds were found to be competitive inhibitors of TPL, with K(i) values of 3.4 mM and 135 microM for 3- and 2-aza-L tyrosine, respectively. Thus, 3-azatyrosine does not act as a transition state analogue, possibly due to the lack of tetrahedral geometry at C-1. However, 2-aza L-tyrosine is the most potent competitive inhibitor of TPL found to date. The K(i) value of 2-aza-L-tyrosine is half that of 2-aza-DL-tyrosine, indicating that the D-enantiomer is inactive as an inhibitor. Neither azatyrosine isomer was shown to be a substrate for beta-elimination, based on coupled assays with lactate dehydrogenase and on HPLC measurements. Both isomers of azatyrosine form equilibrium mixtures of external aldimine and quinonoid intermediates when they bind to TPL. However, 2-azatyrosine reacts about 10-fold faster to form a quinonoid intermediate than does 3-azatyrosine. Since 2-azatyrosine is in the zwitterion or phenolate ion form at all the pH values examined, the strong binding of this compound suggests that L-tyrosine may be bound to the active site of TPL as the phenolate anion. PMID- 11732907 TI - Specific surface association of avidin with N-biotinylphosphatidylethanolamine membrane assemblies: effect on lipid phase behavior and acyl-chain dynamics. AB - The interaction of avidin with aqueous dispersions of N biotinylphosphatidylethanolamines, of acyl chain lengths C(14:0), C(16:0), and C(18:0), was studied by using spin-label electron spin resonance (ESR) spectroscopy, (31)P nuclear magnetic resonance ((31)P NMR) spectroscopy, differential scanning calorimetry, and chemical binding assays. In neutral buffer containing 1 M NaCl, binding of avidin is due to specific interaction with the biotinyl lipid headgroup because avidin presaturated with biotin does not bind. Saturation binding of the protein corresponds to a ratio of 50 lipid molecules per tetrameric avidin. Phospholipid probes spin-labeled at various positions between C-4 and C-14 in the sn-2 chain were used to characterize the effects of avidin binding on the lipid chain dynamics. In the fluid phase, protein binding results in a decrease of chain mobility at all positions of labeling while the flexibility gradient characteristic of a liquid-crystalline lipid phase is maintained. There is no evidence from the spin-label ESR spectra for penetration of the protein into the hydrophobic interior of the membrane. At temperatures corresponding to the gel phase, the lipid chain mobility increases on binding protein. The near constancy in mobility found with chain position, however, suggests that in the gel phase the lipid chains remain interdigitated upon binding avidin. Binding of increasing amounts of avidin results in a gradual decrease of the lipid chain-melting transition enthalpy with only small change in the transition temperature. At saturation binding, the calorimetric enthalpy is reduced to zero. (31)P NMR spectroscopy indicates that protein binding increases the surface curvature of dispersions of all three biotin lipids. The C(14:0) biotin lipid yields isotropic (31)P NMR spectra in the presence of avidin at all temperatures between 10 and 70 degrees C, in contrast to dispersions of the lipid alone, which give lamellar spectra at low temperature that become isotropic at the chain-melting temperature. In the presence of avidin, the C(16:0) and C(18:0) biotin lipids yield primarily lamellar (31)P NMR spectra at low temperature with a small isotropic component; the intensity of the isotropic component increases with temperature, and the spectra narrow and become totally isotropic at high temperature, in contrast to dispersions of the lipids alone, which give lamellar spectra in the fluid phase. The binding of avidin therefore reduces the cooperativity of the biotin lipid packing, regulates the mobility of the lipid chains, and enhances the surface curvature of the lipid aggregates. These effects may be important for both lateral and transbilayer communication in the membrane. PMID- 11732908 TI - Dual effect of ATP in the activation mechanism of brain Ca(2+)/calmodulin dependent protein kinase II by Ca(2+)/calmodulin. AB - The activation mechanism of Ca(2+)/calmodulin-dependent protein kinase II (alphaCaMKII) is investigated by steady-state and stopped-flow fluorescence spectroscopies. Lys(75)-labeled TA-cal [Torok, K., and Trentham, D. R. (1994) Biochemistry 33, 12807-12820] is used to measure binding events, and double labeled AEDANS,DDP-T34C/T110/C-calmodulin [Drum et al. (2000) J. Biol. Chem. 275, 36334-36340] (DA-cal) is used to detect changes in calmodulin conformation. Fluorescence quenching of DA-cal attributed to resonance energy transfer is related to the compactness of the calmodulin molecule. Interprobe distances are estimated by lifetime measurements of Ca(2+)/DA-cal in complexes with unphosphorylated nucleotide-free, nucleotide-bound, and Thr(286)-phospho alphaCaMKII as well as with alphaCaMKII-derived calmodulin-binding peptides in the presence of Ca(2+). These measurements show that calmodulin can assume at least two spectrally distinct conformations when bound to alphaCaMKII with estimated interprobe distances of 40 and 22-26 A. Incubation with ATP facilitates the assumption of the most compact conformation. Nonhydrolyzable ATP analogues partially replicate the effects of ATP, suggesting that while the binding of ATP induces a conformational change, Thr(286)-autophosphorylation is probably required for the transition of calmodulin into its most compact conformer. The rate constant for the association of Ca(2+)/TA-cal with alphaCaMKII is estimated as 2 x 10(7) M(-1) s(-1) and is not substantially affected by the presence of ATP. The rate of net calmodulin compaction measured by Ca(2+)/DA-cal is markedly slower, occurring with a rate constant of 2.5 x 10(6) M(-1) s(-1), suggesting that unproductive complexes may play a role in the activation mechanism. PMID- 11732909 TI - Effects of phospholipid headgroup and phase on the activity of diacylglycerol kinase of Escherichia coli. AB - Diacylglycerol kinase (DGK) of Escherichia coli has been reconstituted into a variety of phospholipid bilayers and its activity determined as a function of lipid headgroup structure and phase preference. The anionic phospholipids dioleoylphosphatidic acid, dioleoylphosphatidylserine, and cardiolipin were all found to support activities lower than that supported by dioleoylphosphatidylcholine. In mixtures of dioleoylphosphatidylcholine and 20 mol % anionic phospholipids, the presence of anionic phospholipids all resulted in lower activities than in dioleoylphosphatidylcholine, except for dioleoylphosphatidylglycerol whose presence had little effect on activity. In some cases, the low activity in the presence of anionic phospholipid followed from a decrease in v(max); in some cases, it followed from an increase in the K(m) for diacylglycerol, and in the case of dioleoylphosphatidic acid, it followed from both. Activities in mixtures containing 80 mol % dioleoylphosphatidylethanolamine were lower than in dioleoylphosphatidylcholine at temperatures where both lipids adopted a bilayer phase; at higher temperatures where dioleoylphosphatidylethanolamine preferred a hexagonal H(II) phase, the differences in activity were greater. These experiments suggest that the presence of lipids preferring a hexagonal H(II) phase leads to low activities. Activities of DGK are low in a gel phase lipid. PMID- 11732910 TI - Ultrastructural and biochemical localization of N-RAP at the interface between myofibrils and intercalated disks in the mouse heart. AB - N-RAP is a recently discovered muscle-specific protein found at cardiac intercalated disks. Double immunogold labeling of mouse cardiac muscle reveals that vinculin is located immediately adjacent to the fascia adherens region of the intercalated disk membrane, while N-RAP extends approximately 100 nm further toward the interior of the cell. We partially purified cardiac intercalated disks using low- and high-salt extractions followed by density gradient centrifugation. Immunoblots show that this preparation is highly enriched in desmin and junctional proteins, including N-RAP, talin, vinculin, beta1-integrin, N cadherin, and connexin 43. Electron microscopy and immunolabeling demonstrate that N-RAP and vinculin are associated with the large fragments of intercalated disks that are present in this preparation, which also contains numerous membrane vesicles. Detergent treatment of the partially purified intercalated disks removed the membrane vesicles and extracted vinculin and beta1-integrin. Further separation on a sucrose gradient removed residual actin and myosin and yielded a fraction morphologically similar to fasciae adherentes that was highly enriched in N-RAP, N-cadherin, connexin 43, talin, desmin, and alpha-actinin. The finding that N-RAP copurifies with detergent-extracted intercalated disk fragments even though beta-integrin and vinculin have been completely removed suggests that N RAP association with the adherens junction region is mediated by the cadherin system. Consistent with this hypothesis, we found that recombinant N-RAP fragments bind alpha-actinin in a gel overlay assay. In addition, immunofluorescence shows that N-RAP remains bound at the ends of isolated, detergent-treated cardiac myofibrils. These results demonstrate that N-RAP remains tightly bound to myofibrils and fasciae adherentes during biochemical purification and may be a key constituent in the mechanical link between these two structures. PMID- 11732911 TI - Specific photoaffinity-labeling of Tyr-50 on the heavy chain and of Tyr-32 on the light chain in the steroid combining site of a mouse monoclonal anti-estradiol antibody using C3-, C6-, and C7-linked 5-azido-2-nitrobenzoylamidoestradiol photoreagents. AB - A mouse monoclonal anti-7-(O-carboxymethyl)oximinoestradiol antibody 9D3, raised against the same immunogen as that employed for generating the reported anti estradiol antibody 15H11 [Rousselot, P., et al. (1997) Biochemistry 36, 7860 7868], was found to exhibit an opposite specificity profile with a much stronger recognition of the D-ring than of the A-ring extremity of the steroid, but a similar lack of specificity for both 6- and 7-positions of the B-ring. This antibody was photoaffinity-labeled with five (5-azido-2-nitrobenzoyl)amido (ANBA) derivatives of [17alpha-(3)H]estradiol, synthesized from 3-aminoethyloxy, 3 (aminoethylamido)carboxymethyloxy, 6alpha- and 6beta-amino, and 7-[O (aminoethylamido)carboxymethyl]oximino precursors. After tryptic digestion, the radioactive peptides on L and H chains were immunopurified with the immobilized antibody 9D3, separated by reversed-phase liquid chromatography, sequenced, and characterized by mass spectrometry, including post-source decay-matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The long 3-(ANBA ethylamido)carboxymethyl ether photoreagent was found to label TyrL-32 (on CDR L1), whereas no labeling was observed with the shorter 3-derivative, a result in agreement with a binding pocket large enough to explain the high cross-reactivity with estradiol 3-conjugates. The two 6alpha- and 6beta-ANBA-estradiol isomers, as well as the 7-[O-(ANBA-ethylamido)carboxymethyl]oximinoestradiol photoreagent derived from the steroid hapten, labeled the same TyrL-32 residue. The 6beta-ANBA epimer also labeled TyrH-50 (at the basis of CDR H2). These experiments indicate that TyrL-32 is freely accessible from the three C3, C6, and C7 positions, all presumed to be exposed to solvent, while TyrH-50 is probably located on the beta face of estradiol. These results, obtained in solution, provide experimental data useful for molecular modeling of the steroid-antibody complex. PMID- 11732912 TI - Endothelial cell PAF synthesis following thrombin stimulation utilizes Ca(2+) independent phospholipase A(2). AB - Platelet activating factor (PAF) is a potent lipid autocoid that is rapidly synthesized and presented on the surface of endothelial cells following thrombin stimulation. PAF production may occur via de novo synthesis or by the combined direct action of phospholipase A(2) (PLA(2)) and acetyl-CoA:lyso-PAF acetyltransferase or via the remodeling pathway. This study was undertaken to define the role of PLA(2) and plasmalogen phospholipid hydrolysis in PAF synthesis in thrombin-treated human umbilical artery endothelial cells (HUAEC). Basal PLA(2) activity in HUAEC was primarily found to be Ca(2+)-independent (iPLA(2)), membrane-associated, and selective for arachidonylated plasmenylcholine substrate. Thrombin stimulation of HUAEC resulted in a preferential 3-fold increase in membrane-associated iPLA(2) activity utilizing plasmenylcholine substrates with a minimal increase in activity with alkylacyl glycerophospholipids. No change in cystolic iPLA(2) activity in thrombin stimulated HUAEC was observed. The thrombin-stimulated activation of iPLA(2) and associated hydrolysis of plasmalogen phospholipids was accompanied by increased levels of arachidonic acid (from 1.1 +/- 0.1 to 2.8 +/- 0.1%) and prostacyclin release (from 38 +/- 12 to 512 +/- 24%) as well as an increased level of production of lysoplasmenylcholine (from 0.6 +/- 0.1 to 2.1 +/- 0.3 nmol/mg of protein), lysophosphatidylcholine (from 0.3 +/- 0.1 to 0.6 +/- 0.1 nmol/mg of protein), and PAF (from 790 +/- 108 to 3380 +/- 306 dpm). Inhibition of iPLA(2) with bromoenol lactone resulted in inhibition of iPLA(2) activity, plasmalogen phospholipid hydrolysis, production of choline lysophospholipids, and PAF synthesis. These data indicate that PAF production requires iPLA(2) activation in thrombin-stimulated HUAEC and may occur through the CoA-independent transacylase remodeling pathway rather than as a direct result of the PLA(2)-catalyzed hydrolysis of membrane alkylacyl glycerophosphocholine. PMID- 11732913 TI - Electron-transfer reactions of the reductase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath). AB - Soluble methane monooxygenase (sMMO) catalyzes the hydroxylation of methane by dioxygen to afford methanol and water, the first step of carbon assimilation in methanotrophic bacteria. This enzyme comprises three protein components: a hydroxylase (MMOH) that contains a dinuclear nonheme iron active site; a reductase (MMOR) that facilitates electron transfer from NADH to the diiron site of MMOH; and a coupling protein (MMOB). MMOR uses a noncovalently bound FAD cofactor and a [2Fe-2S] cluster to mediate electron transfer. The gene encoding MMOR was cloned from Methylococcus capsulatus (Bath) and expressed in Escherichia coli in high yield. Purified recombinant MMOR was indistinguishable from the native protein in all aspects examined, including activity, mass, cofactor content, and EPR spectrum of the [2Fe-2S] cluster. Redox potentials for the FAD and [2Fe-2S] cofactors, determined by reductive titrations in the presence of indicator dyes, are FAD(ox/sq), -176 +/- 7 mV; FAD(sq/hq), -266 +/- 15 mV; and [2Fe-2S](ox/red), -209 +/- 14 mV. The midpoint potentials of MMOR are not altered by the addition of MMOH, MMOB, or both MMOH and MMOB. The reaction of MMOR with NADH was investigated by stopped-flow UV-visible spectroscopy, and the kinetic and spectral properties of intermediates are described. The effects of pH on the redox properties of MMOR are described and exploited in pH jump kinetic studies to measure the rate constant of 130 +/- 17 s(-)(1) for electron transfer between the FAD and [2Fe-2S] cofactors in two-electron-reduced MMOR. The thermodynamic and kinetic parameters determined significantly extend our understanding of the sMMO system. PMID- 11732914 TI - The role of redox-active amino acids on compound I stability, substrate oxidation, and protein cross-linking in yeast cytochrome C peroxidase. AB - The role of two tryptophans (Trp51 and Trp191) and six tyrosines (Tyr36, Tyr39, Tyr42, Tyr187, Tyr229, and Tyr236) in yeast cytochrome c peroxidase (CcP) has been probed by site-directed mutagenesis. A series of sequential mutations of these redox-active amino acid residues to the corresponding, less oxidizable residues in lignin peroxidase (LiP) resulted in an increasingly more stable compound I, with rate constants for compound I decay decreasing from 57 s(-1) for CcP(MI, W191F) to 7 s(-1) for CcP(MI, W191F,W51F,Y187F,Y229F,Y236F,Y36F,Y39E,Y42F). These results provide experimental support for the proposal that the stability of compound I depends on the number of endogenous oxidizable amino acids in proteins. The higher stability of compound I in the variant proteins also makes it possible to observe its visible absorption spectroscopic features more clearly. The effects of the mutations on oxidation of ferrocytochrome c and 2,6-dimethoxyphenol were also examined. Since the first mutation in the series involved the change of Trp191, a residue that plays a critical role in the electron transfer pathway between CcP and cyt c, the ability to oxidize cyt c was negligible for all mutant proteins. On the other hand, the W191F mutation had little effect on the proteins' ability to oxidize 2,6-dimethoxyphenol. Instead, the W51F mutation resulted in the largest increase in the k(cat)/K(M), from 2.1 x 10(2) to 5.0 x 10(3) M(-1) s(-1), yielding an efficiency that is comparable to that of manganese peroxidase (MnP). The effect in W51F mutation can be attributed to the residue's influence on the stability and thus reactivity of the ferryl oxygen of compound II, whose substrate oxidation is the rate-determining step in the reaction mechanism. Finally, out of all mutant proteins in this study, only the variant containing the Y36F, Y39E, and Y42F mutations was found to prevent covalent protein cross-links in the presence of excess hydrogen peroxide and in the absence of exogenous reductants. This finding marks the first time a CcP variant is incapable of forming protein cross-links and confirms that one of the three tyrosines must be involved in the protein cross-linking. PMID- 11732915 TI - FTIR studies of phytochrome photoreactions reveal the C=O bands of the chromophore: consequences for its protonation states, conformation, and protein interaction. AB - The molecular changes of phytochrome during red --> far-red and reverse photoreactions have been monitored by static infrared difference spectroscopy using the recombinant 65 kDa N-terminal fragment assembled with a chromophore chemically modified at ring D or with a chromophore isotopically labeled with (18)O at the carbonyl group of ring A. This allows the identification of the C=O stretching vibrations of rings D and A. We exclude the formation of an iminoether in Pfr. The positions of both these modes show that the chromophore always remains protonated. The upshift of the C=O stretch of ring D in the first photoproducts is explained by a twisted methine bridge connecting rings C and D. The changes in the vibrational pattern during the red --> far-red conversion show that the backreaction is not just the reversal of the forward reaction. The infrared difference spectra of the fragment deviate very little from those of the full-length protein. The differences which are related to the lack of the C terminal half of the protein constituting the signaling domain are possibly important for the understanding of the signaling mechanism. PMID- 11732916 TI - Specific DNA recognition by the type II restriction endonuclease MunI: the effect of pH. AB - To investigate the effect of pH on sequence-specific binding, a thermodynamic characterization of the interaction of the protein MunI with a specific, and a nonspecific, oligonucleotide was performed. MunI is a type II restriction endonuclease which is able to bind specifically, but loses its enzymatic activity in the absence of magnesium ions. Comparison of the specific and nonspecific interactions at 10 and 25 degrees C shows that the latter is accompanied by a small change in enthalpy, and a negligible change in constant pressure heat capacity. On going through the pH range 5.75-9.0 at 25 degrees C, the affinity of specific complex formation is reduced by 20-fold. The interaction is accompanied by the protonation of groups assumed to be on the protein. Based on the simplest model that will fit the data, two distinct protonation events are observed. At low pH, two groups per protein molecule undergo protonation with a pK(a) of 6.0 and 6.9 in the free and bound forms, respectively. At high pH, a further independent protonation occurs involving two groups with pK(a) values of 8.9 and approximately 10.7 in the free and bound forms, respectively. The change in heat capacity ranges from -2.7 to -1.7 kJ mol(-1) K(-1) in going from pH 6.5 to 8.5. This range of variation of change in heat capacity can be accounted for by the effects of protonation of the interacting molecules. The change in heat capacity, calculated from surface area burial using a previously established relationship (1.15 kJ mol(-1) K(-1)), does not correlate well with the experimentally determined values. PMID- 11732917 TI - Effect of sequence context on O(6)-methylguanine repair and replication in vivo. AB - Understanding the origins of mutational hotspots is complicated by the intertwining of several variables. The selective formation, repair, and replication of a DNA lesion, such as O(6)-methylguanine (m(6)G), can, in principle, be influenced by the surrounding nucleotide environment. A nearest neighbor analysis was used to address the contribution of sequence context on m(6)G repair by the Escherichia coli methyltransferases Ada or Ogt, and on DNA polymerase infidelity in vivo. Sixteen M13 viral genomes with m(6)G flanked by all permutations of G, A, T, and C were constructed and individually transformed into repair-deficient and repair-proficient isogenic cell strains. The 16 genomes were introduced in duplicate into 5 different cellular backgrounds for a total of 160 independent experiments, for which mutations were scored using a recently developed assay. The Ada methyltransferase demonstrated strong 5' and 3' sequence specific repair of m(6)G in vivo. The Ada 5' preference decreased in the general order: GXN > CXN > TXN > AXN (X = m(6)G, N = any base), while the Ada 3' preference decreased in the order: NX(T/C) > NX(G/A), with mutation frequencies (MFs) ranging from 35% to 90%. The Ogt methyltransferase provided MFs ranging from 10% to 25%. As was demonstrated by Ada, the Ogt methyltransferase repaired m(6)G poorly in an AXN context. When both methyltransferases were removed, the MF was nearly 100% for all sequence contexts, consistent with the view that the replicative DNA polymerase places T opposite m(6)G during replication irrespective of the local sequence environment. PMID- 11732918 TI - Intrinsic double-stranded-RNA processing activity of Escherichia coli ribonuclease III lacking the dsRNA-binding domain. AB - The ribonuclease III superfamily represents a structurally related group of double-strand (ds) specific endoribonucleases which play key roles in diverse prokaryotic and eukaryotic RNA maturation and degradation pathways. A dsRNA binding domain (dsRBD) is a conserved feature of the superfamily and is important for substrate recognition. RNase III family members also exhibit a "catalytic" domain, in part defined by a set of highly conserved amino acids, of which at least one (a glutamic acid) is important for cleavage but not for substrate binding. However, it is not known whether the catalytic domain requires the dsRBD for activity. This report shows that a truncated form of Escherichia coli RNase III lacking the dsRBD (RNase III[DeltadsRBD]) can accurately cleave small processing substrates in vitro. Optimal activity of RNase III[DeltadsRBD] is observed at low salt concentrations (<60 mM Na(+)), either in the presence of Mg(2+) (>25 mM) or Mn(2+) ( approximately 5 mM). At 60 mM Na(+) and 5 mM Mn(2+) the catalytic efficiency of RNase III[DeltadsRBD] is similar to that of RNase III at physiological salt concentrations and Mg(2+). In the presence of Mg(2+) RNase III[DeltadsRBD] is less efficient than the wild-type enzyme, due to a higher K(m). Similar to RNase III, RNase III[DeltadsRBD] is inhibited by high concentrations of Mn(2+), which is due to metal ion occupancy of an inhibitory site on the enzyme. RNase III[DeltadsRBD] retains strict specificity for dsRNA, as indicated by its inability to cleave (rA)(25), (rU)(25), or (rC)(25). Moreover, dsDNA, ssDNA, or an RNA-DNA hybrid are not cleaved. Low (micromolar) concentrations of ethidium bromide block RNase III[DeltadsRBD] cleavage of substrate, which is similar to the inhibition seen with RNase III and is indicative of an intercalative mode of inhibition. Finally, RNase III[DeltadsRBD] is sensitive to specific Watson-Crick base-pair substitutions which also inhibit RNase III. These findings support an RNase III mechanism of action in which the catalytic domain (i) can function independently of the dsRBD, (ii) is dsRNA specific, and (iii) participates in cleavage site selection. PMID- 11732919 TI - Zinc fingers and thiol-disulfide oxidoreductase activities of chaperone DnaJ. AB - Chaperone DnaJ is a homodimer with each subunit containing 10 cysteine residues and two Zn(II) ions, which have been identified to form two zinc fingers, C(144)DVC(147)Zn(II)C(197)NKC(200) (Zn1) and C(161)PTC(164)Zn(II)C(183)PHC(186) (Zn2), with C(265) and C(323) in reduced form. Guanidine hydrochloride at 6.4 M destroys only Zn1, which does not reform after refolding. p Hydroxymercuriphenylsulfonate acid, but not ethylenediaminetetraacetic acid (EDTA) even at high concentrations, can remove two Zn(II) ions from DnaJ, but only Zn2 can be reconstituted. After removal of Zn(II) ions, only C(144) and C(147) in Zn1 are oxidation-resistant, and the other six cysteines are easily oxidizable. DnaJ shows reductase activity and oxidase activity but little, if any, isomerase activity. The reductase activity is reversibly inhibited by EDTA. Zn2 is important for the enzymatic activity, and only -C(183)PHC(186)- among the four motifs of -CXXC- functions as the active site of the enzyme. A C-terminal (Q(181)-R(376)) fragment shows a zinc finger of C(183)PHC(186)Zn(II)C(197)NKC(200) and full enzymatic activity of DnaJ. The N terminal half sequence (M(1)-Q(180)) and Zn1 are not required for the enzymatic activity but are important for the chaperone activity of DnaJ. PMID- 11732920 TI - Melatonin reverses the profibrillogenic activity of apolipoprotein E4 on the Alzheimer amyloid Abeta peptide. AB - Inheritance of apoE4 is a strong risk factor for the development of late-onset sporadic Alzheimer's disease (AD). Several lines of evidence suggest that apoE4 binds to the Alzheimer Abeta protein and, under certain experimental conditions, promotes formation of beta-sheet structures and amyloid fibrils. Deposition of amyloid fibrils is a critical step in the development of AD. We report here that addition of melatonin to Abeta in the presence of apoE resulted in a potent isoform-specific inhibition of fibril formation, the extent of which was far greater than that of the inhibition produced by melatonin alone. This effect was structure-dependent and unrelated to the antioxidant properties of melatonin, since it could be reproduced neither with the structurally related indole N acetyl-5-hydroxytryptamine nor with the antioxidants ascorbate, alpha-tocophenol, and PBN. The enhanced inhibitory effects of melatonin and apoE were lost when bovine serum albumin was substituted for apoE. In addition, Abeta in combination with apoE was highly neurotoxic (apoE4 > apoE3) to neuronal cells in culture, and this activity was also prevented by melatonin. These findings suggest that reductions in brain melatonin, which occur during aging, may contribute to a proamyloidogenic microenvironment in the aging brain. PMID- 11732921 TI - Folding of a disulfide-bonded protein species with free thiol(s): competition between conformational folding and disulfide reshuffling in an intermediate of bovine pancreatic ribonuclease A. AB - The conformational folding of the nativelike intermediate des-[40-95] on the major oxidative folding pathway of bovine pancreatic ribonuclease A (RNase A) has been examined at various pHs and temperatures in the absence of a redox reagent. Des-[40-95] has three of the four disulfide bonds of native RNase A and lacks the bond between Cys40 and Cys95. This three-disulfide species was unfolded at low pH to inhibit any disulfide reshuffling and was refolded at higher pH, allowing both conformational folding and disulfide-reshuffling reactions to take place. As a result of this competition, 15-85% of des-[40-95], depending on the experimental conditions, undergoes intramolecular disulfide-reshuffling reactions. That portion of the des-[40-95] population which has native isomers of essential proline residues appears to fold faster than the disulfide reaction can occur. However, when the folding is retarded, conceivably by the presence of non-native isomers of essential proline residues, des-[40-95] may reshuffle before completing the conformational folding process. These results enable us to distinguish among current models for the critical structure-forming step in oxidative folding and reveal a new model for coupling proline isomerization to disulfide-bond formation. These experiments also demonstrate that the reshuffling folding competition assay is a useful tool for detecting structured populations in conformational folding intermediates. PMID- 11732922 TI - Product release during the first turnover of the ATP sulfurylase-GTPase. AB - ATP sulfurylase, from Escherichia coli Kappa-12, is a GTPase target complex that catalyzes and couples the chemical potentials of two reactions: GTP hydrolysis and activated sulfate (APS) synthesis. Previous work suggested that the product release branch of the GTPase mechanism might include rate-determining release and/or isomerization step(s). Such steps are known to couple chemical potentials in other energy transducing systems. Rate-determining, product release step(s) were confirmed in the ATP sulfurylase-GTPase reaction by a burst of product in pre-steady-state, rapid-quench experiments. Classical rapid-quench experiments, which measure total product formation, do not allow the slow steps to be assigned to the release of a specific product, or to slow isomerization, because they do not distinguish solution-phase from enzyme-bound product. Assay systems that exclusively monitor solution-phase P(i) and GDP were used to obtain free product progress curves during the first turnover of ATP sulfurylase. Together, the free and total product data describe how the products partition between the enzyme surface and solution during the first turnover. In combination, the data provide the time dependence of the concentrations of specific product intermediates, AMP.PP(i).E.GDP.P(i) and AMP.PP(i).E.GDP, the rate constants for the release of P(i) (4.2 s(-1)) and GDP (4.8 s(-1)) from these complexes, respectively, and the equilibrium constant for the enzyme-bound, beta,gamma-bond cleavage reaction: [AMP.PP(i).E.GTP']/[AMP.PP(i).E.GDP.P(i)] = 0.7. The data are fit, using global analysis, to obtain a complete kinetic and energetic description of this GTPase reaction. PMID- 11732923 TI - Probing the general base catalysis in the first step of BamHI action by computer simulations. AB - BamHI is a type II restriction endonuclease that catalyzes the scission of the phoshodiester bond in the GAGTCC cognate sequence in the presence of two divalent metal ions. The first step of the reaction is the preparation of water for nucleophilic attack by Glu-113, which has been proposed to abstract the proton from the attacking water molecule. Alternatively, the 3'-phosphate group to the susceptible phosphodiester bond has been suggested to play a role as the general base. The two hypotheses have been tested by computer simulations using the semiempirical protein dipoles Langevin dipoles (PDLD/S) method. Deprotonation of water by Glu-113 has been found to be less favorable by 5.7 kcal/mol than metal catalyzed deprotonation with a concomitant proton transfer to bulk solvent. The preparation of the nucleophile by the 3'-phosphate group is less favorable by 12.3 kcal/mol. These results suggest that both the general base and the substrate assisted mechanisms in the first step of BamHI action are less likely than the metal-catalyzed reaction. The metal ions in the active site of BamHI make the largest contributions to the reduction of the free energy of hydroxide ion formation. On the basis of these findings we propose that the first step of endonuclease catalysis does not require a general base; rather, the essential attacking nucleophile in BamHI catalytic action is stabilized by the metal ions. PMID- 11732924 TI - Characterization of the tryptophan residues of Escherechia coli alkaline phosphatase by phosphorescence and optically detected magnetic resonance spectroscopy. AB - The phosphorescence and zero field optically detected magnetic resonance (ODMR) of the tryptophan (Trp) residues of alkaline phosphatase from Escherechia coli are examined. Each Trp is resolved optically and identified with the aid of the W220Y mutant and the terbium complex of the apoenzyme. Trp(109), known from earlier work to be the source of room-temperature phosphorescence (RTP), emits a highly resolved low-temperature phosphorescence (LTP) spectrum and has the narrowest ODMR bands observed thus far from any protein site, revealing a uniquely homogeneous local environment. The decay kinetics of Trp(109) at 1.2 K reveals that the major triplet population (70%) undergoes inefficient crystallike spin-lattice relaxation by direct interaction with lattice phonons, the remainder being relaxed efficiently by local disorder modes. The latter population is smaller than is typical for protein sites, suggesting an unusual degree of local rigidity and order consistent with the long-lived RTP. Trp(220) emits a broader LTP spectrum originating to the blue of Trp(109). It has typically broad ODMR bands consistent with local heterogeneity. The LTP of Trp(268) has an ill-defined origin blue shifted relative to Trp(220) and ODMR frequencies consistent with a greater degree of solvent exposure. Trp(268) has noticeable dispersion of its decay kinetics, consistent with quenching at the triplet level by a nearby disulfide residue. PMID- 11732925 TI - Serine 232 of the alpha(2A)-adrenergic receptor is a protein kinase C-sensitive effector coupling switch. AB - alpha(2)-adrenergic receptors (alpha(2)AR) couple to multiple effectors including adenylyl cyclase and phospholipase C. We hypothesized that signaling selectivity to these effectors is dynamically directed by kinase-sensitive domains within the third intracellular loop of the receptor. Substitution of Ala for Ser232, which is in the N-terminal region of this loop in the alpha(2A)AR, resulted in a receptor that was markedly uncoupled ( approximately 82% impairment) from stimulation of inositol phosphate accumulation while the capacity to inhibit adenylyl cyclase remained relatively intact. In S232A alpha(2A)AR transfected cell membranes, agonist-promoted [(35)S]GTPgammaS binding was reduced by approximately 50%. Coexpression of modified G proteins rendered insensitive to pertussis toxin revealed that the S232A receptor was uncoupled from both G(i) and G(o). S232 is a potential PKC phosphorylation site, and whole cell phosphorylation studies showed that the mutant had depressed phosphorylation compared to wild type (1.3- vs 2.1-fold/basal). Consistent with S232 directing coupling to phospholipase C, PMA exposure resulted in approximately 67% desensitization of agonist-promoted inositol phosphate accumulation without significantly affecting inhibition of adenylyl cyclase. The dominant effect of mutation or phosphorylation at this site on inositol phosphate as compared to cAMP signaling was found to most likely be due to the low efficiency of signal transduction via phospholipase C vs adenylyl cyclase. Taken together, these results indicate that S232 acts as a selective, PKC-sensitive, modulator of effector coupling of the alpha(2A)AR to inositol phosphate stimulation. This represents one mechanism by which cells route stimuli directed to multifunctional receptors to selected effectors so as to attain finely targeted signaling. PMID- 11732926 TI - Activation of protein kinase C alpha by lipid mixtures containing different proportions of diacylglycerols. AB - Lipid activation of protein kinase C alpha (PKC alpha) was studied using a model mixture containing POPC/POPS (molar ratio 4:1) and different proportions of either DPG or POG. The lipid mixtures containing DPG were physically characterized by using different physical techniques, and a phase diagram was constructed by keeping a constant POPC/POPS molar ratio of 4:1 and changing the concentration of 1,2-DPG. The phase diagram displayed three regions delimited by two compounds: compound 1 (CO(1)) with 35 mol % of 1,2-DPG and compound 2 (CO(2)) with 65 mol % of 1,2-DPG. PKC alpha activity was assayed at increasing concentrations of 1,2-DPG, maximum activity being reached at 30 mol % 1,2-DPG, which decreased at higher concentrations. Maximum activity occurred, then, at concentrations of 1,2-DPG which corresponded to the transition from region 1 to region 2 of the phase diagram. It was interesting that this protein was maximally bound to the membrane at all DPG concentrations. Similar results were observed when the enzyme was activated by POG, when a maximum was reached at about 10 mol %. This remained practically constant up to 50 mol %, about which it decreased, the binding level remaining maximal and constant at all POG concentrations. The fact that in the assay conditions used maximal binding was already reached even in the absence of diacylglycerol was attributed to the interaction of the C2 domain with the POPS present in the membrane through the Ca(2+) ions also present. To confirm this, the isolated C2 domain was used, and it was also found to be maximally bound at all DPG concentrations and even in its absence. Since the intriguing interaction patterns observed seemed to be due then to the C1 domain, the PKC alpha mutant D246/248N was used. This mutant has a decreased Ca(2+)-binding capacity through the C2 domain and was not activated nor bound to membranes by increasing concentrations of DPG. However, POG was able to activate the mutant, which showed a similar dependence on POG concentration with respect to activity and binding to membranes. These data underline the importance of unsaturation in one of the fatty acyl chains of the diacylglycerol. PMID- 11732927 TI - FTIR and resonance Raman studies of nitric oxide binding to H93G cavity mutants of myoglobin. AB - Nitric oxide (NO) binds to the myoglobin (Mb) cavity mutant, H93G, forming either a five- or six-coordinate Fe-NO complex. The H93G mutation eliminates the covalent attachment between the protein and the proximal ligand, allowing NO to bind H93G possibly from the proximal side of the heme rather than the typical diatomic binding pocket on the distal side. The question of whether NO binds on the distal or proximal side was addressed by FTIR spectroscopy of the N-O vibrational frequency nuN(-O) for a set of Mb mutants that perturb the electrostatic environment of the heme pocket. Vibrational spectra of five- and six-coordinate MbNO complexes indicate that nu(N-O) shifts (by as much as 26 cm( 1)) to higher energies for the distal mutants H64V and H64V/H93G relative to the energies of wild-type and H93G MbNO, while nu(N-O) is not affected by the proximal side mutation S92A/H93G. This result suggests that NO binds on the distal side of heme in the five- and six-coordinate MbNO complexes of H93G. Additionally, values of the Fe-NO vibrational frequency nu(Fe-NO) as measured by resonance Raman spectroscopy are reported for the distal and proximal double mutants of H93G. These results suggest that nu(Fe-NO) is not very sensitive to mutations that perturb the electrostatic environment of the heme pocket, leading to the observation that nu(N-O) and nu(Fe-NO) are not quantitatively correlated for the MbNO complexes presented here. Furthermore, nu(N-O) and nu(Fe-NO) do not correlate well with equilibrium constants for imidazole binding to the five coordinate MbNO complexes of the H93G double mutants. The data presented here do not appear to support the presence of pi-back-bonding or an inverse trans effect of NO binding in Mb mutants that alter the electrostatic environment of the heme pocket. PMID- 11732928 TI - White-coat hypertension--to treat or not to treat: a clinical dilemma. PMID- 11732929 TI - The real contribution of the major risk factors to the coronary epidemics: time to end the "only-50%" myth. PMID- 11732930 TI - A practical approach to achieving recommended blood pressure goals in diabetic patients. AB - Approximately 11 million Americans have both hypertension and diabetes mellitus. This double diagnosis places such patients at high risk for renal damage, especially end-stage renal disease. The sixth report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure recommends a blood pressure goal of less than 130/85 mm Hg to reduce or slow the onset of renal disease and cardiovascular events in patients with hypertension and diabetes mellitus. Recent data, however, now suggest that an even lower diastolic blood pressure goal (ie, <80 mm Hg) may be necessary. Studies have shown that use of angiotensin-converting enzyme inhibitors can prevent the progression of microalbuminuria to overt proteinuria, reduce proteinuria in patients with overt diabetic nephropathy, slow the deterioration of the glomerular filtration rate, delay progression to end-stage renal disease, and lower blood pressure. Thus, all diabetic patients with blood pressure greater than 130/80 mm Hg should begin angiotensin-converting enzyme inhibitor treatment and be titrated to moderate or high doses until the blood pressure goal is achieved. However, monotherapy still may not control blood pressure to the recommended target. Studies have shown that use of multiple antihypertensive agents is necessary and successful in helping patients reach their target blood pressure, and this may offer more renoprotection than one agent used singly. A case study that applies these concepts in outpatient practice is included. PMID- 11732931 TI - Septic thrombosis of the cavernous sinuses. AB - Septic thrombosis of the cavernous sinuses (or cavernous sinus thrombophlebitis [CST]) is a dramatic and potentially lethal illness, which is still occasionally seen by clinicians. Before the availability of antimicrobial agents, mortality from CST was near 100%, but it markedly decreased to approximately 20% to 30% during the antibiotic era. Nevertheless, the threat of death and serious morbidity continues to necessitate early recognition, diagnosis, and treatment of CST to minimize risks to the patient. Accordingly, we reviewed the salient clinical features of this illness, with emphasis on newer aspects of diagnosis and treatment. PMID- 11732932 TI - Left ventricular changes in isolated office hypertension: a blood pressure matched comparison with normotension and sustained hypertension. AB - BACKGROUND: Isolated office (IO) hypertension is a benign condition according to some researchers, whereas others believe it is associated with cardiovascular abnormalities and increased cardiovascular risk. The aim of this study is to compare morphofunctional characteristics of the left ventricle (LV) in IO hypertensive subjects, normotensive subjects (hereafter, hypertensives and normotensives), and never-treated sustained hypertensives. The 3 groups were matched not only by age, sex, and body mass index but also by clinic blood pressure (BP) (IO hypertensives and sustained hypertensives) and daytime BP (IO hypertensives and normotensives). METHODS: We enrolled 42 IO hypertensives (clinic BP > 140 and/or 90 mm Hg and daytime BP < or = 130/80 mm Hg), 42 sustained hypertensives (clinic BP > 140 and/or 90 mm Hg and daytime BP > or = 140 and/or 90 mm Hg) and 42 normotensives (clinic BP < 135 and/or 85 mm Hg and daytime BP < or = 130/80 mm Hg). Left ventricular morphologic features and function were assessed using digitized M-mode echocardiography. RESULTS: Compared with normotensives, IO hypertensives had significantly thicker LV walls, increased LV mass, reduced diastolic function, increased prevalence of LV hypertrophy, and preclinical diastolic dysfunction. Sustained hypertensives, compared with IO hypertensives, had significantly thicker LV wall, higher LV mass, and lower diastolic function, whereas the prevalence of LV hypertrophy and preclinical diastolic dysfunction was greater than in IO hypertensives, but the difference did not reach statistical significance (P = .29). CONCLUSIONS: Comparing matched BP groups, IO hypertensives have LV morphofunctional characteristics considerably different from normotensives and qualitatively similar to sustained hypertensives. Therefore, our results support the hypothesis that IO hypertension should not be considered as simply a benign condition. PMID- 11732933 TI - Total cholesterol/HDL cholesterol ratio vs LDL cholesterol/HDL cholesterol ratio as indices of ischemic heart disease risk in men: the Quebec Cardiovascular Study. AB - BACKGROUND: Total cholesterol (TC)/high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C)/HDL-C ratios are used to predict ischemic heart disease risk. There is, however, no consensus on which of these 2 indices is superior. The objective of the present study was to present evidence that the LDL-C/HDL-C ratio may underestimate ischemic heart disease risk in overweight hyperinsulinemic patients with high triglyceride (TG)-low HDL-C dyslipidemia. METHODS: A total of 2103 middle-aged men in whom measurements of the metabolic profile were performed in the fasting state were recruited from 7 suburbs of the Quebec metropolitan area. RESULTS: The relationship of LDL-C/HDL-C to TC/HDL-C ratios was examined among men in the Quebec Cardiovascular Study classified into tertiles of fasting TG levels. For any given LDL-C/HDL-C ratio, the TC/HDL-C ratio was higher among men in the top TG tertile (>168 mg/dL [>1.9 mmol/L]) than in men in the first and second TG tertiles. Adjustment of the TC/HDL-C ratio for LDL-C/HDL-C by covariance analysis generated significant differences in average TC/HDL-C ratios among TG tertiles (P<.001). Greater differences in features of the insulin resistance syndrome (insulinemia, apolipoprotein B, and LDL size) were noted across tertiles of the TC/HDL-C ratio than tertiles of the LDL-C/HDL-C ratio. CONCLUSION: Variation in the TC/HDL-C ratio may be associated with more substantial alterations in metabolic indices predictive of ischemic heart disease risk and related to the insulin resistance syndrome than variation in the LDL-C/HDL-C ratio. PMID- 11732934 TI - Successful aging in the oldest old: Who can be characterized as successfully aged? AB - BACKGROUND: Successful aging is a worldwide aim, but it is less clear which indicators characterize elderly persons as successfully aged. We explored the meaning of successful aging from 2 perspectives. METHODS: Analysis of data from the first cross-sectional part of the longitudinal Leiden 85-plus Study, conducted in Leiden, the Netherlands. All inhabitants of Leiden aged 85 years were eligible. Data were obtained from 599 participants (response rate, 87%). Successful aging from a public health perspective was defined as a state of being. All participants were classified as successful or not successful based on optimal scores for physical, social, and psychocognitive functioning and on feelings of well-being, using validated quantitative instruments. Qualitative indepth interviews on the perspectives of elderly persons were held with a representative group of 27 participants. RESULTS: Although 45% (267/599) of the participants had optimal scores for well-being, only 13% (79/599) had optimal scores for overall functioning. In total, 10% (58/599) of the participants satisfied all the criteria and could be classified as successfully aged. The qualitative interviews showed that most elderly persons viewed success as a process of adaptation rather than a state of being. They recognized the various domains of successful aging, but valued well-being and social functioning more than physical and psychocognitive functioning. CONCLUSIONS: If successful aging is defined as an optimal state of overall functioning and well-being, only a happy few meet the criteria. However, elderly persons view successful aging as a process of adaptation. Using this perspective, many more persons could be considered to be successfully aged. PMID- 11732935 TI - Generalist and subspecialist physicians' knowledge, attitudes, and practices regarding influenza and pneumococcal vaccinations for elderly and other high-risk patients: a nationwide survey. AB - BACKGROUND: Influenza and pneumococcal vaccination rates remain too low. This survey assessed generalist and subspecialist physicians' knowledge, attitudes, and practices regarding influenza and pneumococcal vaccinations for high-risk patients. METHODS: A self-administered questionnaire was mailed to 6000 physicians randomly selected from a national database. RESULTS: After 3 mailings, 1874 physicians (32%) of the 5858 eligible responded. Although most physicians thought that it was very important for their high-risk patients be current on influenza and pneumococcal vaccinations, only 86% and 75% of generalists and subspecialists, respectively, very strongly recommended influenza vaccinations to their elderly patients and only 81% and 64%, respectively, very strongly recommended pneumococcal vaccinations to their elderly patients (P<.001 for both). After multivariate logistic regression, factors significantly associated with strongly recommending vaccinations to elderly patients in the influenza and pneumococcal vaccination models included female sex of provider, the provider having received an influenza vaccination, the provider's beliefs about vaccine effectiveness and cost-effectiveness, a patient's risk for illness, and ease of targeting patients. In addition, generalists were more likely than subspecialists to strongly recommend pneumococcal vaccinations to their patients. Patient reminders, special clinics, and standing orders were each used by fewer than 30% of respondents, although generalists were more likely than subspecialists to use such strategies. CONCLUSIONS: Nontrivial proportions of generalist and subspecialist physicians fail to strongly recommend influenza and pneumococcal vaccinations to their elderly and high-risk patients. Other effective strategies for promoting vaccine delivery are also used relatively infrequently. These findings suggest areas for improvement if vaccination rates are to reach national goals. PMID- 11732936 TI - Severity assessment of lower respiratory tract infection in elderly patients in primary care. AB - BACKGROUND: Simple markers for evaluating the severity of lower respiratory tract infections (LRTI) in primary care are lacking. It is of value to examine whether the information available to the primary care physician during a patient's initial visit can be used to assess the severity of LRTI. METHODS: The associations between different baseline variables and outcomes (survival within or more than 30 days) were investigated prospectively in a series of 950 home living patients 65 years or older with severe LRTI that their primary care physicians suspected to be pneumonia. RESULTS: Twenty-one men and 17 women died (4.1%) within 30 days. According to univariate analysis, the following parameters differed (P < .01) between the fatalities and survivors: acute aggravation of a coexisting illness, age, respiratory rate, white blood cell count, and C-reactive protein (CRP) level. According to Cox forward stepwise regression analysis (P = .01 for entry and.05 for removal), acute aggravation of a concurrent illness, respiratory rate (> or = 25/min), and CRP concentration (> or = 100 mg/L) were independently associated with death. The mortality rate was 2.2% if the patients had none or only 1 of the independent risk factors and 20% if they had all 3 risk factors. CONCLUSIONS: Preceding aggravation of a concurrent illness and respiratory rate of 25/min or higher, together with an elevated serum CRP level (> or = 100 mg/L), can be used as simple markers for identifying patients with the highest risk for LRTI and improve management decisions among elderly people in primary care. PMID- 11732937 TI - The effects of estrogen replacement therapy on airway function in postmenopausal, asthmatic women. AB - BACKGROUND: Data from multiple clinical, epidemiologic, and in vitro studies are conflicting regarding the effect of estrogen replacement therapy (ERT) on airway function in postmenopausal women with asthma. OBJECTIVE: To determine the impact of withdrawal of estrogen administration in postmenopausal, asthmatic women. METHODS: Twenty asthmatic women who were postmenopausal for at least 2 years and undergoing ERT were recruited for this prospective crossover study. Subjects continued taking baseline estrogen for 28 days, stopped taking estrogen for 28 days, and then resumed taking the medication for 14 days. Objective measurements were obtained by recording daily peak flows in the morning and evening and formal spirometry at days 14, 28, 42, 56, and 70. Compliance was measured by evaluating serum estradiol levels at days 28 and 56. Daily use of short-acting beta-agonist bronchodilators was also recorded. RESULTS: Differences in estradiol levels indicated compliance with the medication regimen. The combined day 14 and 28 (taking estrogen) mean percent predicted forced expiratory volume in 1 second (FEV(1)) was 77% compared with the combined day 42 and 56 (not taking estrogen) mean FEV(1) of 78% and the day 70 (taking estrogen again) FEV(1) of 76% (P>.05). Average peak flow measurements were 295.5 L/min for the duration of ERT, 293.9 L/min while not undergoing ERT, and 291.8 L/min when ERT was restarted for the final 2 weeks of the study (P>.05). Use of short-acting beta-agonist bronchodilators did not differ between study periods. CONCLUSION: These data indicate that neither the discontinuation nor reinitiation of ERT in postmenopausal, asthmatic women has any effect on objective measures of airway obstruction. PMID- 11732938 TI - Pain management in frail, community-living elderly patients. AB - BACKGROUND: Pain is a common problem among older people living in different community settings. As indicated by the World Health Organization (WHO), pain can be relieved using pharmacologic agents. However, pain continues to be addressed inadequately. OBJECTIVES: To describe the prevalence of pain in frail elderly people living in the community and to evaluate the adequacy of pain management. METHODS: We analyzed data from a large collaborative observational study group, the Italian Silver Network Home Care project, that collected data on patients admitted to home health care programs. Twelve home health care agencies participated in the project evaluating the implementation of the Minimum Data Set for Home Care instrument. We enrolled 3046 patients, 65 years and older, in the present study. The main outcome measures were the prevalence of daily pain and analgesic treatment. RESULTS: A total of 1341 individuals (39%, 49%, and 41% of those aged 65-74, 75-84, and > or = 85 years, respectively) reported daily pain. Of patients with daily pain, 25% received a WHO level 1 drug; 6%, a WHO level 2 drug; and 3%, a WHO level 3 drug (eg, morphine sulfate). Patients 85 years or older were less likely to receive analgesics compared with the younger patients (univariate odds ratio, 0.73; 95% confidence interval [CI], 0.60-0.89). Another independent predictor of failing to receive any analgesic was low cognitive performance (adjusted odds ratio, 0.80; 95% CI, 0.69-0.93). CONCLUSIONS: Daily pain is prevalent among frail elderly patients living in the community and is often untreated, particularly among older and demented patients. PMID- 11732939 TI - Economic burden of pneumonia in an employed population. AB - OBJECTIVE: To estimate the overall economic burden of pneumonia from an employer perspective. METHODS: The annual, per capita cost of pneumonia was determined for beneficiaries of a major employer by analyzing medical, pharmaceutical, and disability claims data. The incremental costs of 4036 patients with a diagnosis of pneumonia identified in a health claims database of a national Fortune 100 company were compared with a 10% random sample of beneficiaries in the employer overall population. RESULTS: Total annual, per capita, employer costs were approximately 5 times higher for patients with pneumonia ($11 544) than among typical beneficiaries in the employer overall population ($2368). The increases in costs were for all components (eg, medical care, prescription drug, disability, and particularly for inpatient services). A small proportion (10%) of pneumonia patients (almost all of whom were hospitalized) accounted for most (59%) of the costs. CONCLUSIONS: Patients with pneumonia present an important financial burden to employers. These patients use more medical care services, particularly inpatient services, than the average beneficiary in the employer overall population. In addition to direct health care costs related to medical utilization and the use of prescription drugs, indirect costs due to disability and absenteeism also contribute to the high cost of pneumonia to an employer. PMID- 11732940 TI - Malignant ascites: new concepts in pathophysiology, diagnosis, and management. AB - Malignant ascites is a manifestation of advanced malignant disease that is associated with significant morbidity. Mainstays of treatment include diuretics and recurrent large volume paracentesis. Although lymphatic obstruction has been considered the major pathophysiologic mechanism behind its formation, recent evidence suggests that immune modulators, vascular permeability factors, and metalloproteinases are contributing significantly to the process. These new observations offer the opportunity for development of new, more targeted therapies for the treatment of malignant ascites. This article uses a clinical case to highlight the problem, then reviews these new concepts in the pathophysiology of malignant ascites formation. The diagnosis and management of this challenging medical problem are subsequently discussed, with emphasis on how these new pathophysiologic insights are being applied to the development of novel therapies that may soon change how we manage this troubling clinical condition. PMID- 11732941 TI - Vertical transmission of HIV-1 variants resistant to reverse transcriptase and protease inhibitors. PMID- 11732942 TI - Evaluation of adherence to highly active antiretroviral therapy. PMID- 11732944 TI - Child development in pediatrics: beyond rhetoric. PMID- 11732945 TI - The impact of medical school loans and the promise of loan repayment on entry into a pediatric academic career. PMID- 11732946 TI - To be rather than not to be--that is the problem with the questions we ask adolescents about their childbearing intentions. AB - OBJECTIVE: To demonstrate that rephrasing the questions used to assess childbearing intentions to quantify the strength of the intent to remain nonpregnant, rather than the strength of the intent to become pregnant, would make teenagers' responses more useful to health care providers, family planning counselors, and health policy makers. METHODS: Examples from the teen pregnancy prevention literature are used to support the recommendations for change. RESULTS: Teenagers rarely plan their pregnancies. However, because those who are having sexual intercourse must actively try not to become pregnant or they will likely conceive, teenagers often become pregnant because they lack a firm commitment not to do so. Thus, to accurately profile the antecedents of adolescent pregnancy, (1) the questions used to assess childbearing intentions must be rephrased so that teenagers who intend to remain nonpregnant can be distinguished from those who do not and (2) separate differential diagnoses must be developed for inconsistent contraceptive use within these 2 groups of teenagers who are at risk for unintended pregnancy. CONCLUSION: Asking sexually active teenagers about the strength of their intent to remain nonpregnant will make the results of office interviews and national surveys more useful because the responses such questions elicit will enable health care providers and policy makers to target common, modifiable antecedents of inconsistent contraceptive use for interventions. PMID- 11732947 TI - Predictors of bacterial meningitis in the era after Haemophilus influenzae. AB - OBJECTIVE: To determine if, in the era after Haemophilus influenzae type b, the cerebrospinal fluid (CSF) white blood cell (WBC) count can be safely used to stratify children suspected of having bacterial meningitis into low- and high risk groups. DESIGN: Retrospective analysis of CSF samples. SETTING: Tertiary care pediatric center in Toronto, Ontario, between January 1, 1992, and October 1, 1996. PATIENTS: All CSF samples collected on children aged 2 months to 17 years were included. The final database consisted of 1617 atraumatic samples from children without prior neurologic or immunologic disease who underwent a lumbar puncture to assess the possibility of community-acquired bacterial meningitis. MAIN OUTCOME MEASURES: The predictive values of CSF WBC count, differential, protein, and glucose. RESULTS: There were 44 cases of bacterial meningitis. Five had 3 CSF WBCs per microliter or less, and 6 had 4 to 30 CSF WBCs per microliter. The negative predictive value of CSF specimens with 30 WBCs per microliter or less for bacterial meningitis was 99.3%. Cerebrospinal fluid samples with greater than 30 WBCs per microliter had a likelihood ratio for bacterial meningitis of 10.3 (95% confidence interval, 8.0-13.1) and a positive predictive value of 22.3%. Other significant predictors of bacterial meningitis included age, CSF glucose, protein, gram stain, CSF-serum glucose ratio, and peripheral blood band count. CONCLUSIONS: Given the occurrence of bacterial meningitis in children in the absence of CSF pleocytosis, other factors should be considered when managing children with suspected bacterial meningitis. Children older than 6 months with 30 CSF WBCs per microliter or less are at low risk for bacterial meningitis. If clinically stable and without other laboratory markers of bacterial meningitis, hospital admission and empiric antibiotic therapy may be unwarranted. PMID- 11732948 TI - Diagnosing bacterial meningitis after the Haemophilus influenzae vaccine: still a challenge. PMID- 11732949 TI - Primary care services promoting optimal child development from birth to age 3 years: review of the literature. AB - OBJECTIVE: To examine the evidence base for primary health care services promoting the optimal development of typically developing children aged birth to 3 years. DATA SOURCES: Peer-reviewed publications addressing clinical evaluations of primary care services from the MEDLINE and PsychINFO databases. STUDY SELECTION: Criteria for selection were as follows: (1) publication between 1979 and 1999; (2) evaluation of efficacy or effectiveness of education, intervention, and care coordination services or validation of assessment approaches; (3) services applicable to an office practice setting; (4) target population including children aged birth to 3 years; and (5) publication in English. DATA EXTRACTION: A total of 312 publications were abstracted and reviewed by both of us; 47 were selected for inclusion. DATA SYNTHESIS: Assessments of parental concerns and psychosocial risk factors using validated approaches seem to be more accurate in identifying developmental problems than clinicians' appraisals. In general, study results support the efficacy of (1) primary care educational efforts toward promoting optimal parent-child interaction, parents' understanding of child temperament, book-sharing activities, and approaches to healthy sleep habits and (2) office interventions such as counseling for the management of excessive infant crying and sleep problems. CONCLUSIONS: The literature suggests that many primary care activities promoting the optimal development of children are efficacious. Evaluations of developmental assessment and services in primary care should be expanded in depth and breadth. The implications for clinical practice, training, and health care policy are discussed. PMID- 11732950 TI - Inpatient care for uncomplicated bronchiolitis: comparison with Milliman and Robertson guidelines. AB - CONTEXT: Bronchiolitis is the most common lower respiratory tract infection in infancy. A recent Centers for Disease Control and Prevention report confirmed that hospitalization rates for bronchiolitis have increased 2.4-fold from 1980 to 1996. Controversies exist about optimal treatment plans. Milliman and Robertson recommend ambulatory care management; in case of hospitalization, the recommended length of stay is 1 day. OBJECTIVES: To relate actual practice variation for infants admitted with uncomplicated bronchiolitis to Milliman and Robertson's recommendations. DESIGN: Prospective observational study. SETTING: General care wards of 8 pediatric hospitals of the Child Health Accountability Initiative during the winter of 1998-1999. PATIENTS: First-time admissions for uncomplicated bronchiolitis in patients not previously diagnosed as having asthma and who were younger than 1 year. MAIN OUTCOME MEASURES: Respiratory rate, monitored interventions, attainment of discharge criteria goals, and length of stay. RESULTS: Eight hundred forty-six patients were included in the final analysis: 85.7% were younger than 6 months, 48.5% were nonwhite, and 64.1% were Medicaid recipients or self-pay. On admission to the hospital, 18.3% of the infants had respiratory rates higher than higher than 80 breaths per minute, 53.8% received supplemental oxygen therapy, and 52.6% received intravenous fluids. These proportions decreased to 1.9%, 33.8%, and 20.3%, respectively, 1 day after admission, and to 0.7%, 20.1%, and 8.6%, respectively, 2 days after admission. The average length of stay was 2.8 days (SD, 2.3 days). CONCLUSIONS: Milliman and Robertson's recommendations do not correspond to practice patterns observed at the hospitals participating in this study; no hospital met the Milliman and Robertson recommended 1-day goal length of stay. Administration of monitored intervention persisted past the second day of hospitalization. PMID- 11732951 TI - Randomized trial of the addition of ipratropium bromide to albuterol and corticosteroid therapy in children hospitalized because of an acute asthma exacerbation. AB - OBJECTIVE: To determine whether the addition of inhaled ipratropium bromide to inhaled albuterol and systemic corticosteroid therapy was more efficacious than inhaled albuterol and systemic corticosteroids alone in the inpatient treatment of acute asthma exacerbations in children. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: Pediatric inpatient unit of a tertiary urban hospital. PARTICIPANTS: Eighty children (aged 1-18 years) hospitalized because of an acute asthma exacerbation. INTERVENTION: Children were randomized to receive either nebulized ipratropium bromide, 250 microg, or nebulized isotonic sodium chloride solution, 1 mL. All children received albuterol and systemic corticosteroids. MAIN OUTCOME MEASURES: The primary outcome variable was a validated clinical asthma score, measured at baseline and every 6 hours for 36 hours. Secondary outcome measures included the forced expiratory volume in 1 second, the oxygen saturation, the number of doses of inhaled study drug, the time to an inhaled drug-dosing interval of 4 hours, and the length of the hospital stay. RESULTS: There were no differences between groups on baseline characteristics. The intention-to-treat analysis, using repeated-measures analysis of variance, showed no significant (P =.07) difference between the groups in the clinical asthma score over time. There were also no significant differences between groups on secondary outcomes. CONCLUSION: The addition of nebulized ipratropium bromide to nebulized beta(2)-agonist and corticosteroid therapy in the treatment of children hospitalized because of asthma (following intensive emergency department treatment) confers no extra benefit. PMID- 11732952 TI - Metered-dose inhaler: the emergency department orphan. AB - OBJECTIVE: Physicians providing emergency department care to children primarily use nebulizers for the delivery of bronchodilators and these physicians have misconceptions regarding the advantages and disadvantages of using metered-dose inhalers (MDIs) with a spacer (MDI + S) for acute asthma exacerbations. DESIGN: Self-administered mail survey. SETTING: Emergency department. PARTICIPANTS: Emergency medicine section members of the American Academy of Pediatrics and Canadian Pediatric Society. INTERVENTIONS: Bronchodilator delivery methods in acute pediatric asthma. MAIN OUTCOME MEASURES: The 2 principal outcomes for bivariate analysis were self-reported nebulizer use in all patients and MDI + S use in patients with mild acute asthma. RESULTS: Of eligible physicians, 333 (51%) of 567 responded. The majority were dual trained in pediatrics and pediatric emergency medicine (72%) and practiced full time (83%) in an urban (83%) pediatric emergency department (80%). The most commonly cited advantages of MDIs were their cost (33%) and speed of use (28%). The most commonly cited disadvantages were patient or parent dissatisfaction (24%) and relative ease of nebulizer use (23%). Only 10% to 21% of participants used MDIs in the emergency department and reserved this delivery method for children with mild asthma exacerbations. There were no significant associations between selected respondent demographic variables and the use of MDIs. CONCLUSIONS: Misconceptions regarding the efficacy and safety of MDI + S for the treatment of acute asthma exacerbations exist but are limited to a minority of surveyed emergency medicine physicians caring for children. Nebulizers remain the preferred method of routine bronchodilator delivery by physicians providing care to pediatric asthmatics in the emergency department. PMID- 11732953 TI - Effectiveness of oral or nebulized dexamethasone for children with mild croup. AB - OBJECTIVE: To assess the efficacy of oral dexamethasone or nebulized dexamethasone sodium phosphate in children with mild croup. METHODS: Double blind, placebo-controlled study of 264 children between 6 months and 6 years of age with symptoms of croup for fewer than 48 hours. Patients were excluded if they received racemic epinephrine or corticosteroid treatment. Other exclusion criteria included corticosteroid treatment during the 14 days prior to enrollment or complicating medical condition. Subjects randomly received oral dexamethasone (0.6 mg/kg), nebulized dexamethasone sodium phosphate (160 microg), or placebo. Telephone follow-up was obtained on days 1, 2, 3, 4, and 7. MAIN OUTCOME MEASURES: The primary outcome measure was treatment failure, defined as receiving corticosteroid or racemic epinephrine treatment during the 7 days after enrollment in the study. Secondary outcome measures included seeking additional care and the parental assessments of the patients' condition obtained during follow-up (worse, same, better, or gone). RESULTS: Eighty-five patients received oral dexamethasone, 91 received nebulized dexamethasone, and 88 received placebo. There were 3 treatment failures in the oral dexamethasone-treated group, 12 in the nebulized dexamethasone-treated group, and 10 in the placebo-treated group (P =.05). Ten children in the oral dexamethasone-treated group sought additional care compared with 27 and 29 in the nebulized dexamethasone-treated and placebo treated groups, respectively (P =.002). Parents of children in the oral dexamethasone-treated group reported greater improvement on day 1 (P<.001) compared with the nebulized dexamethasone-treated and placebo-treated groups. CONCLUSIONS: Children with mild croup who receive oral dexamethasone treatment are less likely to seek subsequent medical care and demonstrate more rapid symptom resolution compared with children who receive nebulized dexamethasone or placebo treatment. PMID- 11732954 TI - Child health status and parental employment. AB - OBJECTIVE: To understand the relationship between several measures of child health status and the employment of parents. DESIGN: A cross-sectional study using 1994 National Health Interview Survey on Disability data. PARTICIPANTS: A nationally representative sample of children and their parents. OUTCOME MEASURES: Maternal and paternal employment (measured separately). INTERVENTION: We use a series of logistic regression models with maternal and paternal employment as the dependent variables and the health status of the child with the poorest health status in the family as the primary independent variable. Models additionally include sociodemographic correlates of employment. RESULTS: Having a child with poor health status, as measured by general reported health, hospitalizations, activity limitations, and chronic condition or disability status, is associated with reduced employment of mothers and fathers. For example, the odds ratios of being employed for having a child with an activity limitation are 0.75 for mothers (95% confidence interval, 0.67-0.85) and 0.66 for fathers (95% confidence interval, 0.53-0.82). CONCLUSIONS: Having a child with poor health status is associated with reduced maternal and paternal employment. Further studies are needed to determine whether poor child health status causes reductions in parental labor force participation. If such a causal relationship exists, it has important implications for social policy, employment policy, and clinical anticipatory guidance. PMID- 11732955 TI - Exposure to violence: psychological and academic correlates in child witnesses. AB - BACKGROUND: Inner-city children are frequently exposed to violence; however, there are few data regarding the psychological and academic correlates of such exposure in young children at school entry. OBJECTIVES: To document exposure to violence in inner-city children aged 7 years; assess their feelings of distress; and evaluate the relationships of exposure to violence with school performance, behavior, and self-esteem. SETTING: A study center in an inner-city hospital. PARTICIPANTS: One hundred nineteen inner-city children evaluated at age 7 years; 119 caregivers (biological and foster). DESIGN: As part of a longitudinal study, children were administered the following by a masked examiner: Things I Have Seen and Heard (TISH) to assess exposure to violence; Levonn, a cartoon-based interview for assessing children's distress symptoms; and the Culture-Free Self Esteem Inventory, Second Edition. School performance was assessed by school reports and child behavior by the Child Behavior Checklist (CBCL), the Parent Report Form, and the Teacher Report Form. Caregivers for children were administered the parent report version of the Checklist of Children's Distress Symptoms (CCDS-PRV) as well as the CBCL Parent Report Form. MAIN OUTCOME MEASURES: Exposure to violence (TISH); feelings of distress (Levonn); school performance; behavior (CBCL Parent Report Form and CBCL Teacher Report Form); and self-esteem (Culture-Free Self-Esteem Inventory). RESULTS: We found that these children were frequently exposed to violence. For example, 75% had heard gun shots, 60% had seen drug deals, 18% had seen a dead body outside, and 10% had seen a shooting or stabbing in the home (TISH). Many showed signs of depression and anxiety; eg, 61% worried some or a lot of the time that they might get killed or die and 19% sometimes wished they were dead (Levonn). Higher exposure to violence (TISH Total Violence score) was correlated with higher Levonn composite scores for depression and anxiety and with lower self-esteem (P< or =.04), and was also associated with lower grade point average and more days of school absence (P< or =.02). Caregiver assessment of child anxiety correlated poorly with child report of anxiety (P =.58). CONCLUSIONS: Young inner-city children have a high exposure to violence by age 7 years; many show signs of distress that frequently are not recognized by caregivers. Further, higher exposure to violence in children correlates with poorer performance in school, symptoms of anxiety and depression, and lower self-esteem. PMID- 11732956 TI - Persistence of maternal concerns surrounding neonatal jaundice: an exploratory study. AB - OBJECTIVES: To explore whether mothers currently express concerns about neonatal jaundice and perceive it as a serious condition; if so, to identify factors influencing these perceptions; and to elicit maternal recommendations for improved health care interactions. DESIGN: Ethnographic interviews using grounded theory methods. Audiotaped data were transcribed and analyzed for themes using a qualitative data analysis software program. SETTING: University and community hospitals. PARTICIPANTS: Forty-seven Spanish- and English-speaking breastfeeding mothers of otherwise healthy infants with a diagnosis of neonatal jaundice and treated in inpatient and/or outpatient settings. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Qualitative descriptions of maternal experiences with neonatal jaundice. RESULTS: Mothers continued to voice concerns about jaundice and perceive it as serious. They expressed misconceptions, wished to have jaundice explained further, and offered suggestions to improve communications with medical staff. Guilt was common, with mothers believing that they had caused the jaundice. Mothers voiced alarm about the yellow skin and discomfort about jaundice management and worried about perceived short- and long-term effects. Maternal perceptions were exacerbated by cultural differences, language barriers, and subtlety of language and its meaning. Key factors in creating perceptions of jaundice as serious included unexpectedness of and lack of knowledge about jaundice, quality of information received, levels of intervention, and prolonged duration of illness and yellow color. Interactions with health care professionals and other mothers with personal experience with jaundice were important mediators in the way mothers reacted to information. CONCLUSION: Practitioners need to address these persisting misconceptions and concerns about neonatal jaundice with mothers. PMID- 11732958 TI - Collaboration with school nurses: improving the effectiveness of tuberculosis screening. AB - OBJECTIVE: To compare tuberculosis skin test (TST) reading rates between children whose tests were read by school nurses following specific requests by physicians and those who relied on their parents to get their tests read, either at school or at the physician's office. DESIGN: A randomized controlled trial. SETTING: An urban hospital-based pediatric practice. PARTICIPANTS: Healthy low-income Hispanic and African American children aged 5 to 17 years whose physicians ordered TSTs at their routine physical examinations. Subjects attended 1 of 68 public schools. Nurses at these schools were willing to read student TSTs, and received instructions about how to read and report the results back to the physician's office. INTERVENTION: Subjects were randomized to a control group (routine TST placement, with no physician-to-school nurse communication) or to an intervention group (routine TST placement, with physician-to-school nurse communication). MAIN OUTCOME MEASURES: Tuberculosis skin test reading rates between the 2 groups were compared. Impediments to TST reading and reporting were investigated. RESULTS: One hundred thirty-four children were enrolled, 54 (40%) in the control group and 80 (60%) in the intervention group. More patients in the intervention group had their TSTs read by 72 hours compared with those in the control group (74 [92%] vs 30 [56%]; P<.001). The low reading rate in the control group was best attributed to communication failures. CONCLUSION: Systematic collaboration with school nurses can increase TST reading rates. PMID- 11732957 TI - Incidence and circumstances of nonfatal firearm-related injuries among children and adolescents. AB - OBJECTIVE: To describe the incidence and circumstances of nonfatal firearm related injuries among children and adolescents treated in US emergency departments. DESIGN: Data were obtained from the Firearm Injury Surveillance Study, 1993-1997; data were collected through medical record review at hospitals participating in the National Electronic Injury Surveillance System. SETTING: The hospitals participating in National Electronic Injury Surveillance System are a stratified probability sample of all US hospitals. MAIN OUTCOME MEASURES: Numbers and population rates for nonfatal firearm-related injuries among children and adolescents younger than 20 years old. RESULTS: An estimated 115,131 (95% confidence interval, 76,769-153,493) children and adolescents were treated for a nonfatal gunshot wound during the study period. The estimated annual rates of injury (per 100,000) were 2.0 (children 0-4 years old), 2.2 (children 5-9 years old), 15.4 (children 10-14 years old), and 106.5 (adolescents 15-19 years old). The ratios of nonfatal to fatal firearm-related injuries were 4.0 (children 0-4 years old), 4.4 (children 5-9 years old), 5.0 (children 10-14 years old), and 4.4 (adolescents 15-19 years old). An additional estimated 103,814 children (95% confidence interval, 69,223-138,405) were shot with a nonpowder firearm (BB or pellet gun). Boys 5 to 9 and 10 to 14 years old had the highest rates of injury related to nonpowder firearms, an estimated 36.2 and 99.8 per 100,000, respectively. Fifty-six percent of those 15 to 19 years old were assault victims. An estimated 48% of children and adolescents with powder firearm-related gunshot wounds and an estimated 4% with nonpowder firearm injuries were admitted to the hospital. CONCLUSIONS: Nonfatal injuries related to powder firearms and nonpowder firearms (BB or pellet guns) are an important source of injury among US children and adolescents. Ongoing surveillance of nonfatal firearm-related injury among children and adolescents is needed. PMID- 11732959 TI - Barriers that impede the adoption of pediatric information technology. AB - BACKGROUND: Information technology (IT) is a critical but underused component of health care. Many factors contribute to the inconsistent adoption of IT. OBJECTIVE: To review the literature to better elucidate barriers that are likely to affect the adoption of IT by pediatric professionals. DATA SOURCES: Manuscripts were found using a MEDLINE search combining the terms medicine, information systems, and technology transfer. I also obtained references cited by relevant articles. Finally, I explored the Internet using http://www.google.com and http://www.northernlight.com. STUDY SELECTION: Articles discussing barriers or factors affecting the adoption of IT were considered for inclusion. Articles unrelated to clinical IT were excluded. DATA SYNTHESIS: A variety of barriers exist that affect the adoption of useful technologies. Situational barriers include challenges imposed by the current national health environment, financial and legal risks associated with technology purchasing and use, and access to technology. The most significant barrier is that pediatric health care practitioners may lack the knowledge or training to use IT effectively. CONCLUSIONS: Although some barriers exist that may be challenging to overcome, other barriers, such as the lack of knowledge about the uses of IT, are imminently solvable. Efforts to overcome these barriers should begin in earnest and should include educating stakeholders in the care of children and adolescents, as well as improving the knowledge about various technologies available to support pediatric and adolescent health care. PMID- 11732960 TI - Radiological case of the month. Subcutaneous fat necrosis of the newborn. PMID- 11732961 TI - Pathological case of the month. Goodpasture disease. PMID- 11732962 TI - Weight charts and well-child care: how the pediatrician became the expert in child health. AB - Weighing children became popular in the 1910s, when public health workers hoped to identify malnourished children based on weight. They measured tens of thousands of children in school halls and church basements, compared their results with standard weight charts, and reported evidence of widespread malnutrition. In the 1920s, physicians argued that a complete medical history and a physical examination, not just weight, were necessary to diagnose malnutrition. By the 1930s, the weight chart had become merely one of the many diagnostic tools used to monitor the health of well children in the physician's office. Weight charts remain an essential part of pediatric practice, but their history is more than a simple tale of scientific progress. This article explores how pediatrics emerged as a primary care specialty in the midst of conflict over the meaning of weight, the professional role of women in medicine, and the pediatrician's preeminence as a child health expert. PMID- 11732963 TI - Pediatric advocacy: more lessons. PMID- 11732964 TI - Sexual orientation is not the issue. PMID- 11732965 TI - Toleration for the Boy Scouts. PMID- 11732966 TI - The World Health Organization oral rehydration solution in pediatric practice: a comment on the methodology. PMID- 11732969 TI - Predicting duration of posttraumatic amnesia (PTA) from early PTA measurements. AB - OBJECTIVES: To determine a set of variables that would reliably predict duration of posttraumatic amnesia (PTA) in patients with traumatic brain injury and to test the efficacy of the model. DESIGN: Simultaneous standard multiple regression analyses. PARTICIPANTS: Two independent samples of patients with traumatic brain injury who were in the early stages of PTA: a test sample (n = 61) and a cross validation sample (n = 25). MAIN OUTCOME MEASURE: The Modified Oxford PTA Scale (MOPTAS) is a 12-item test measuring orientation (8 items) and anterograde memory (4 items). The Galveston Orientation and Amnesia Test (GOAT) was also used on a subset of the test sample. PROCEDURE: Patients were examined daily until they emerged from PTA. RESULTS: A statistically significant model, using three predictor variables, was derived that reliably predicted duration of PTA, accounting for 89% of the variance. A second model, using two predictor variables readily available to the clinician (day posttrauma on which PTA testing began and aggregate PTA scores over the first 5 days of testing) had comparable predictive accuracy. A third model, using GOAT data, was also statistically significant and successfully accounted for 72% of the variance. The MOPTAS model showed excellent application to an independent (validation) sample, with an intraclass correlation coefficient between observed and predicted durations of PTA of 0.95. Regression equations for all three models are provided to enable calculation of the predicted duration of PTA. CONCLUSIONS: These models can be readily applied in clinical practice and will provide clinically useful estimates of the duration of PTA within the first week of testing after admission to rehabilitation. This information will be important in terms of family counseling and planning of rehabilitation programs. PMID- 11732970 TI - Life satisfaction after traumatic brain injury. AB - OBJECTIVE: To investigate correlates of life satisfaction after traumatic brain injury (TBI). DESIGN: Prospective, longitudinal study of patients with TBI studied 1 and 2 years after injury. SETTING: A specialized inpatient TBI rehabilitation unit in a midwestern academic medical center. SUBJECTS: Two hundred eighteen consecutive patients admitted for rehabilitation, at least 14 years of age, with a primary diagnosis of TBI, consented to participate, and interviewed 1 and/or 2 years after injury (112 interviewed both years, 58 at year 1 only, 48 at year 2 only). MAIN OUTCOME MEASURES: Satisfaction With Life Scale. RESULTS: Stepwise multiple regressions accounted for statistically significant, but small, proportions of variance. Not having a preinjury history of substance abuse and having gainful employment at the time of follow-up were associated with higher life satisfaction both 1 and 2 years after injury. Motor independence at rehabilitation discharge was also associated at 1 year. Current social integration and the absence of depressed mood were associated at 2 years. Life satisfaction was relatively stable between years. Change that did occur was associated with marital status and depressed mood 2 years after injury. CONCLUSIONS: Life satisfaction after TBI seems to be related to attaining healthy and productive lifestyles. Future research should investigate other factors that affect life satisfaction to increase prediction and appreciate all influences on subjective well being after TBI. PMID- 11732971 TI - Social, neuroradiologic, medical, and neuropsychologic correlates of sexually aberrant behavior after traumatic brain injury: a controlled study. AB - OBJECTIVE: To identify social, neuroradiological, medical, and neuropsychological correlates of sexually aberrant behavior (SAB) after traumatic brain injury (TBI). DESIGN: A controlled study using a retrospective file review. SETTING: A brain injury unit providing inpatient and outpatient rehabilitation services. PARTICIPANTS: A sample of males (n = 25) exhibiting SABs and a control group (n = 25) matched for gender, severity of injury, age at injury, and time after injury. MAIN OUTCOME MEASURES: A protocol that recorded data on demographic, injury, radiological, medical, and neuropsychological variables. RESULTS: The SAB group had a significantly higher incidence of postinjury psychosocial disturbance in areas of nonsexual crime and failure to return to work than the matched TBI group. There were no significant differences between the two groups in the incidence of premorbid psychosocial disturbance or postinjury radiological, medical, or neuropsychological variables. CONCLUSIONS: The study results caution against simplistic explanations of SAB as the product of damage to the frontal lobe systems or premorbid psychosocial disturbance. Furthermore, the results suggest that a wide-ranging assessment of people with TBI who exhibit SABs is required, because results of neuropsychological examination alone cannot be considered conclusive. Future research into the etiology of SABs could examine additional factors such as lack of insight, lack of empathy, and premorbid history of family dysfunction. PMID- 11732972 TI - Spontaneous facial expressivity in children with acquired brain injury. AB - OBJECTIVES: To investigate spontaneous facial behavior during social interaction in children with acquired brain injury (ABI), and to explore possible links between ABI, facial expressivity, and caregiver-reported social competence. DESIGN: Between-group comparisons using t tests, within-group comparisons using correlational analyses, correlational analyses of independent variables and outcome measures. PARTICIPANTS: Sixteen school-aged children with ABI and 32 normally developing children of comparable age and gender. MAIN OUTCOME MEASURE: Aspects of facial expressivity derived from coding scheme designed for this study. RESULTS: Children with ABI were less expressive overall and tended to shift expressions less often than normally developing children. Several measures of expressivity were correlated with measures of social competence. CONCLUSIONS: Brain injury inhibits production of some aspects of spontaneous facial expression. Results are suggestive of links between facial behavior and social skills. PMID- 11732973 TI - Sibling adjustment to pediatric traumatic brain injury: a case-controlled pilot study. AB - OBJECTIVE: To assess depressive symptoms, self-concept, and behavior in non affected siblings of children with severe pediatric traumatic brain injury (TBI). DESIGN: Cross-sectional study with case controls. SETTING: Children's hospital tertiary care center. PARTICIPANTS: Twelve siblings of children consecutively admitted to an inpatient rehabilitation unit after a severe TBI. Case controls were randomly selected from the sibling's classmates. MAIN OUTCOME MEASURES: The Child Behavior Checklist (CBCL), the Teacher's Report Form of the CBCL (TRF CBCL), the Self-Perception Profile for Children and the Children's Depression Inventory (CDI). RESULTS: No statistical differences were found in depressive symptoms, self-concept, or behavior between the siblings and their classmates 3 to 18 months after injury. Poorer functional outcomes in the child with a TBI were found to correlate significantly with lower self-concept and more symptoms of depression in the siblings. CONCLUSIONS: Further research is needed to evaluate the potential impact on sibling adjustment after pediatric TBI. PMID- 11732974 TI - Balance findings in a child before and after a mild head injury. AB - OBJECTIVE: This case study reviews the preinjury and postinjury balance performance of an 11-year-old child who sustained a mild head injury. DESIGN: A prospective design was used to document balance 5 days before injury and 1, 4, and 12 weeks after injury. OUTCOME MEASURES: The assessments used were the balance subtest of the Bruininks-Oseretsky Test of Motor Proficiency, the Pediatric Clinical Test of Sensory Interaction for Balance, and the Postural Stress Test. RESULTS: One week after the trauma, balance deficits were observed in the three tests. The deficits improved 4 weeks after the injury, and performance remained stable over the following 2 months for two of the evaluations except for the Postural Stress Test performance, which failed to improve in the 3-month period after trauma. CONCLUSIONS: The good recovery of balance skills observed over the first month after injury seems to validate the current activity restrictions imposed on children after a mild head injury. However, the persistence of poor performance in the area of reactions to external perturbations indicates that some deficits may persist beyond what is expected with this population. PMID- 11732975 TI - Ginkgo biloba: applications in traumatic brain injury. PMID- 11732976 TI - The Congressional Brain Injury Task Force. PMID- 11732978 TI - Oocyte control of ovarian follicular development and function in mammals. AB - A new perspective on ovarian follicular development has emerged over the last decade. Whereas the oocyte was previously considered only a passive recipient of developmental signals from oocyte-associated granulosa cells, it is now clear that communication between oocytes and granulosa cells is bidirectional. A complex interplay of regulatory factors governs the development of both types of cell. This interplay is essential not only for oocyte development but also for follicular development, beginning with the initial assembly of the primordial follicle and continuing throughout ovulation. The existence of an oocyte granulosa cell regulatory loop, essential for normal follicular differentiation as well as for the production of an oocyte competent to undergo fertilization and embryogenesis, is proposed. Although gonadotrophins are essential for driving the differentiation of granulosa cell phenotypes, within its sphere of influence, the oocyte is probably the dominant factor determining the direction of differentiation and the function of the granulosa cells associated with it. PMID- 11732979 TI - Potential role of a sperm-derived phospholipase C in triggering the egg activating Ca2+ signal at fertilization. AB - An increase in intracellular Ca2+ at fertilization is the trigger for egg activation in all species that have been studied. Exactly how sperm-egg interaction leads to this Ca2+ increase has not been established. There is increasing support for the hypothesis that the spermatozoon introduces a Ca2+ releasing protein into the egg cytoplasm after gamete membrane fusion. This review discusses the merits of this 'sperm factor' hypothesis and presents evidence indicating that the sperm factor, at least in mammals, consists of a phospholipase C with distinctive properties. This evidence leads us to propose that, after gamete fusion, a sperm-derived phospholipase C causes production of inositol 1,4,5- trisphosphate, which then generates Ca2+ waves from within the egg cytoplasm. PMID- 11732980 TI - Ligand-independent activation of progestin receptors: relevance for female sexual behaviour. AB - Traditionally, steroid receptors were believed to be activated only by ligand binding; however, recent studies indicate that steroid receptors can also be activated by mechanisms that do not require ligand, referred to as ligand independent activation. Specifically, progestin receptors can be activated in vitro and in vivo after treatment with neurotransmitters, such as dopamine, in the absence of progesterone. Furthermore, mating-related stimuli can also lead to ligand-independent activation of progestin receptors in female rat brain. This finding indicates that environmental stimuli can influence steroid receptor dependent processes in the absence of circulating hormone. This review focuses on studies indicating that progestin receptors can be activated in the absence of progesterone to influence neuronal response and sexual behaviour in rodents. PMID- 11732981 TI - Alterations in development of reproductive and endocrine systems of wildlife populations exposed to endocrine-disrupting contaminants. AB - Wildlife and human populations are affected by contaminants in natural settings. This problem has been a growing concern over the last decade with the realization that various environmental chemicals can alter the development and functioning of endocrine organs, cells and target tissues. Documented disruptions or alterations in reproductive activity, morphology or physiology in wildlife populations have been correlated with contaminant-induced modifications in endocrine system functioning. Alterations of the endocrine system are complex, and not limited to a particular organ or molecular mechanism. For instance, contaminants have been shown to (1) act as hormone receptor agonists or antagonists, (2) alter hormone production at its endocrine source, (3) alter the release of stimulatory or inhibitory hormones from the pituitary or hypothalamus, (4) alter hepatic enzymatic biotransformation of hormones, and (5) alter the concentration or functioning of serum-binding proteins, altering free hormone concentrations in the serum. This review focuses on two of these alterations, altered hormone synthesis and hepatic biotransformation, as a number of recent studies indicate that these actions are important components of endocrine disruption in developing organisms. The possible role of contaminants in altering sex determination mechanisms is also examined. PMID- 11732982 TI - Expression of vascular endothelial growth factor (VEGF) receptors in rat corpus luteum: regulation by oestradiol during mid-pregnancy. AB - The aim of this study was to investigate the expression of vascular endothelial growth factor (VEGF) receptors, the fms-like tyrosine kinase (flt-1) and kinase insert domain-containing region (KDR), in corpora lutea obtained at different stages of the oestrous cycle and during pregnancy in rats. Immunohistochemistry revealed that both flt-1 and KDR were localized in luteal cells in addition to vascular endothelial cells, and that the intensity of staining was stronger in pregnant rats than in cyclic rats. Rats undergoing hypophysectomy-hysterectomy on day 12 of pregnancy were treated with oestradiol until day 15 of pregnancy to determine whether oestradiol is involved in expression of flt-1 and KDR mRNA in the corpus luteum during mid-pregnancy. The flt-1 and KDR mRNA contents in the corpus luteum were decreased significantly by hypophysectomy-hysterectomy, and these decreases recovered significantly after oestradiol treatment. Changes in the mass of the corpus luteum and serum progesterone concentrations paralleled the changes in expression of flt-1 and KDR mRNA. Developmental studies indicated that flt-1 and KDR mRNA contents in the corpus luteum were constant until day 15 of pregnancy but decreased significantly on day 21 of pregnancy. In conclusion, both flt-1 and KDR were expressed in luteal cells in addition to vascular endothelial cells, and expression was upregulated by oestradiol during mid pregnancy. flt-1 and KDR may play a role in development of the corpus luteum and in production of progesterone during mid-pregnancy in rats. PMID- 11732983 TI - Embryotoxicity of regressing corpora lutea in ewes. AB - Experiments were performed to test the null hypotheses that embryonic survival is not affected by the presence of regressing corpora lutea in progestogen supplemented ewes, and that the embryotoxic effects of regressing corpora lutea do not act locally on embryos in the uterine horn adjacent to the regressing corpora lutea. In Expt 1, laparotomies were performed on day 4 after mating, and progestogen supplementation was initiated and continued until pregnancy was diagnosed by ultrasonography on day 25. On day 4 after mating, ewes were lutectomized (n = 17) or sham lutectomized (n = 15), and injected (i.m.) with 5 mg PGF2alpha at 8 h intervals for 2 or 3 days. Controls (n = 14) were sham lutectomized and injected with saline as described above. Pregnancy rates did not differ in ewes treated with PGF2alpha for 2 rather than 3 days. Pregnancy rates were lower in ewes treated with PGF 2a compared with controls (P < 0.01). In ewes treated with PGF2alpha, lutectomy resulted in an increase in pregnancy rates (59%) compared with ewes subjected to sham lutectomy (33%; P < 0.05). In Expt 2, progestogen supplementation was initiated in the morning of day 4 after mating and continued until pregnancy diagnosis on day 25. In the afternoon of day 4, one ovary selected at random was lutectomized in ewes (n = 34) with at least one corpus luteum on each ovary, and the uterine horns were isolated by ligation to impede intraluminal transfer of luteal or uterine products that might initiate embryonic death. On days 5-8 after mating, equal numbers of ewes were injected i.m. with either saline or 5 mg PGF2alpha at 8 h intervals. Pregnancy rates did not differ between isolated uterine horns contralateral and ipsilateral to the regressing corpus luteum; however, pregnancy rates were lower in PGF2alpha treated ewes than in saline-treated ewes (34 and 77%; P < 0.05). In conclusion, regressing corpora lutea exert an embryotoxic effect; however, there is no evidence that this effect occurs through systemic pathways. PMID- 11732984 TI - Comparison of the capacitation-like state of cooled boar spermatozoa with true capacitation. AB - Cryopreserved spermatozoa demonstrate reduced conception rates compared with fresh spermatozoa when used for artificial insemination. The preliminary stage of cryopreservation of spermatozoa involves cooling to 5 degrees C, during which spermatozoa experience a capacitation-like change, which may be partially responsible for the reduced conception rate observed. The aim of this study was to determine the nature of these capacitation-like changes and how much this process resembles true capacitation. Boar spermatozoa, cooled to 5 degrees C and re-warmed to physiological temperatures (39 degrees C), were compared with spermatozoa capacitated in Tyrode's complete medium (TALP) for 2 h at 39 degrees C. Fluorescent probes, and SDS-PAGE and western blotting were used to visualize events known to occur during capacitation in vitro. Chlortetracycline staining of membrane domains and Fluo-3 detection of changes in intracellular free calcium by flow cytometry in cooled and re-warmed spermatozoa showed similarities to those of capacitated spermatozoa. Alterations to lipid bilayer fluidity assessed by merocyanine fluorescence staining and intracellular signalling pathways detected by tyrosine phosphorylation of cooled and re-warmed spermatozoa, did not completely reflect the changes detected during capacitation in vitro. Thus, cooling spermatozoa to 5 degrees C results in a similar endpoint to that observed in capacitated cells in terms of reactive membranes and changes in intracellular ion concentrations, which may account for their comparable functionality. However, these modifications are not completely analogous and should not be considered true capacitation, but rather a by-passing of the capacitation process. PMID- 11732985 TI - Relaxin-like factor (RLF) serum concentrations and gubernaculum RLF receptor display in relation to pre- and neonatal development of rats. AB - Deletion of the relaxin-like factor (RLF) gene in mice causes retention of testicles and infertility. The development of a synthetic RLF has made it possible to investigate the events that connect the genomic event and the basic biological responses that cause gonadal positioning. Anti-RLF antibodies were raised against synthetic RLF, allowing determination of RLF concentrations during the critical period, testing for RLF receptors on the gubernaculum and exploration of the temporal relationship between receptor display and migration of the testes in developing rats. In male rat pups, serum RLF concentrations were high at day 2 before parturition (2.4 ng ml(-1)) and decreased sharply just before parturition. Thereafter, males and females had the same low serum concentrations until RLF concen-trations began to increase in males only, starting at day 10 after parturition and continuing until adult RLF concentrations (0.6 ng ml(-1)) were reached on day 39 after parturition. The testicles are descending into the scrotum during this phase of increasing RLF concentrations and are descended fully by day 19-21 after parturition, before adult hormone concentrations are established. The high prenatal serum RLF concentration coincides with high expression of RLF receptors in the gubernaculum tissue. Competitive binding of RLF per mg of membrane protein prepared from rat gubernacula at various developmental stages showed no increase in receptor density as sexual maturity was reached. Gubernaculum cells in primary culture showed an increased uptake of 5-bromo-2'-deoxyuridine in the presence of RLF compared with controls. These studies demonstrate that the synthetic RLF is biologically active and indicate that the cryptorchid phenotype INSL3(-/-) is a direct consequence of defective gubernaculum growth, caused by the absence of RLF during early phases of development. PMID- 11732986 TI - Increased expression of major histocompatibility complex (MHC) class I transplantation antigens in bovine trophoblast cells before fusion with maternal cells. AB - The mammalian fetus is potentially at risk from maternal immune attack because it can express paternally inherited polymorphic antigens, including those encoded by the major histocompatibility complex (MHC). The aim of this study was to investigate in more detail MHC class I upregulation by binucleate trophoblast cells in the bovine placenta. A method was developed to isolate binucleate cells by enzymatic disaggregation and density gradient centrifugation of bovine placental cotyledons. In cytospin preparations, 25-30% of purified binucleate cells stained positively with antibodies that recognize bovine MHC class I. The same antibodies were used to immunoprecipitate radiolabelled class I molecules from lysates of binucleate cells and fetal peripheral blood mononuclear cells. The protein species isolated from the two types of cell were similar in size and degree of glycosylation. PCR amplification of cDNA generated from binucleate cells and subsequent sequence analysis demonstrated transcription of MHC class I mRNA species similar to those found in fetal peripheral blood mononuclear cells, and there was no evidence of genetic imprinting of paternally inherited alleles. These results indicate that binucleate cells upregulate expression of MHC class I as they differentiate from MHC-negative uninucleate trophoblast cells. This finding has important implications for the immunological status of the fetus, as binucleate trophoblast cells are destined to cross to the maternal side of the placenta where they fuse with maternal cells. The immunological function of the resulting antigenically mixed fetomaternal hybrid minisyncytia is unknown. PMID- 11732987 TI - Effect of maternal undernutrition during pregnancy on early ovarian development and subsequent follicular development in sheep fetuses. AB - Gonad development in female sheep fetuses is thought to occur in a number of key stages. The aim of this study was to determine the effects of maternal undernutrition, applied at one or more of these critical stages, on fetal ovarian development. Groups of ewes (n = 11-19) were fed rations providing either 100% (high; H) or 50% (low; L) of energy requirements for live weight maintenance during selected 'windows' during gestation. Control ewes (HH and HHH) were fed the H ration from mating until they were killed at days 50, 65 (HH) or 110 (HHH) of gestation, whereas ewes of other groups were fed the L ration for the periods between day 0 and day 30 of gestation (LH and LHH), day 31 and day 50 or 65 of gestation (HL and HLH), day 65 and day 110 of gestation (HHL) or day 0 of gestation until the animals were killed (LL and LLL). At day 50 of gestation, there was no effect of nutritional treatment on mean fetal mass but compared with HH animals, mean fetal ovarian mass was significantly lower in HL (P < 0.05) and LL (P < 0.001) animals. At day 65 of gestation, there were significantly fewer germ cells (P < 0.05) at the resting, diplotene stage of initial meiosis in LL animals than there were in HH animals, indicating delayed germ cell maturation and onset of meiosis. Qualitative assessment of proliferative cell nuclear antigen immunostaining indicated that, at day 50 of gestation, staining was located predominantly in the germ cells, whereas by day 65 of gestation, staining was confined predominantly to somatic cells. Undernutrition in each one of these windows was associated with delayed ovarian follicular development (P < 0.05 0.001) as measured by development of the granulosa cell layer at day 110 of gestation. This study demonstrates that undernutrition before and during folliculogenesis can delay fetal follicular development. PMID- 11732988 TI - Reduction in reproductive lifespan of tissue inhibitor of metalloproteinase 1 (TIMP-1)-deficient female mice. AB - Tissue inhibitor of metalloproteinase 1 (TIMP-1) is a multifunctional protein expressed in the ovary and uterus of females of several species including mice, rats and humans. Mice deficient in TIMP-1 protein display altered reproductive parameters, which include reduced serum progesterone concentrations and abnormal uterine morphology. As these abnormalities could lead to altered reproductive performance, the objective of this study was to examine whether TIMP-1 deficiency is associated with a reduction in the reproductive lifespan of female mice in which the TIMP-1 gene is disrupted. Wild-type (n = 50) and TIMP-1 null mice (n = 90) were mated with males of proven fertility from the same genotype for 12 months during which the number of litters delivered and the number of pups per litter were determined. Significantly fewer TIMP-1 null females achieved pregnancy (47 of 90 or 52%) compared with the wild-type mice (39 of 50 or 78%; P < 0.05). TIMP-1-deficient female mice that achieved and maintained pregnancy had significantly fewer litters during the 12 months (2.9 +/- 0.8 versus 3.5 +/- 0.8; P < 0.01) and significantly fewer pups per litter (5.7 +/- 1.3 versus 4.7 +/- 1.1; P < 0.05) than did wild-type mice. Females of both genotypes produced two consecutive litters, after which significantly fewer TIMP-1 null mice became pregnant. Mating of females with males of proven fertility from the other genotype confirmed that these abnormalities were not due to the inability of TIMP 1 null males to produce offspring. These data indicate that the absence of TIMP-1 is associated with a reduction in the reproductive lifespan of female mice, which may be manifested at the ovary, uterus or both organs. PMID- 11732989 TI - Calcium-binding proteins from the outer acrosomal membrane of ram spermatozoa: potential candidates for involvement in the acrosome reaction. AB - After an intracellular calcium influx, fusion of the sperm plasma membrane and outer acrosomal membrane (the acrosome reaction) precedes mammalian fertilization in vivo. This study describes the isolation of outer acrosomal membrane from ram spermatozoa and the subsequent characterization of calcium-binding proteins. Pooled ejaculates were diluted, cooled slowly and washed. Incubation with Hyamine 1622 (benzethenium chloride) and subsequent slow centrifugation gently dislodged and concentrated acrosomal membranes, the fragments of which were isolated on a two-step discontinuous sucrose gradient. The acrosomal membrane material stained with Giemsa, whereas spermatozoa from the gradient pellet stained intensely only in the equatorial segment. The acrosomal fraction showed a limited number of polypeptides by SDS-PAGE. Incubation with 45Ca2+ revealed two radioactive bands at 34 and 39 kDa. Extraction in the presence of EGTA implied that these proteins are not peripheral proteins associated with the membrane only in the presence of calcium ions, but are integral membrane proteins. Polyclonal antisera raised to the two bands showed specific binding to the anterior acrosomal region and demonstrated the intracellular location of the proteins. Sequence data of protein A revealed 83% homology with calnexin homologue precursor and 70% homology with annexin XI. Protein B showed 68% homology with protein SP-10 precursor and 64-72% homology with various annexins. However, crossreactivity with a range of commercial annexin antibodies and a specific antibody to a synthetic motif encompassing the annexin calcium-binding site was not demonstrable. It is concluded that the isolated proteins are unlikely to be annexins, but are possibly novel calcium-binding proteins. PMID- 11732990 TI - Effect of nutritional supplementation on quantities of glucose transporters 1 and 4 in sheep granulosa and theca cells. AB - The stimulatory effect of nutritional supplementation on ovarian activity in sheep has been linked to an increase in glucose availability that, with insulin, directly decreases follicular steroidogenesis. Glucose uptake occurs by glucose transporters, but it is not known which glucose transporters are present in the sheep ovary or whether they are affected by nutritional stimulation. The aim of this study was to determine whether widely distributed glucose transporter 1 (GLUT1) or insulin-responsive GLUT4 are present in the granulosa or theca cells of sheep ovarian follicles, and whether their concentrations are affected by nutritional stimulation. Merino ewes (n = 49-51 per group) were stimulated nutritionally for 5 days before luteolysis with lupin grain or with one of two regimens of a glucogenic mixture, administered orally, which increases blood glucose concentrations towards the upper end of the normal range. Water was used as a control. Ovaries (n = 3 per group) were dissected and the granulosa cells and thecal shell from individual follicles were examined for glucose transporters using western blotting. GLUT1 concentration was 7-18 times higher in the granulosa than in the theca cells. GLUT4 was detected at a similar concentration in both types of cell. Nutritional treatment had no effect on the concentration of GLUT1 or GLUT4 in either tissue, and did not increase ovulation rate, despite increased concentrations of glucose and insulin. Concentrations of glucose transporters were not correlated with follicular concentrations of oestradiol or androstenedione. The presence of GLUT1 and GLUT4 in the granulosa and theca of sheep follicles indicates that the transporters have a role within the ovary in the modulation of follicular function. PMID- 11732991 TI - Localization of nitric oxide synthase activity in unfertilized oocytes and fertilized embryos during preimplantation development in mice. AB - Changes in the activities of nitric oxide synthase (NOS) during embryonic development, and the distribution of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) isoforms were examined in unfertilized mouse oocytes at the second meiotic metaphase (MII) stage and in fertilized mouse embryos during preimplantation development. In addition, the effects of NOS inhibitors on mouse preimplantation development in vitro were investigated. The activities of NOS in MII oocytes and fertilized embryos during the preimplantation period were determined by NADPH-diaphorase staining. Although NOS activity was detected in unfertilized MII oocytes, the intensity of staining was much weaker than that of fertilized embryos at the one-cell stage. There was a decrease in NOS activity in embryos from the four-cell to the eight-cell stage; however, NOS activity increased again in embryos at the morula stage, particularly in the inner cell population. In the expanded blastocysts, staining was confined to the inner cell mass. Immuno-cytochemical staining showed that eNOS and iNOS were expressed in the cytoplasm of oocytes and embryos during the preimplantation period, and eNOS was also distributed in the nuclei of the embryos. When one-cell embryos were treated with 1 mmol N(omega)-nitro-L-arginine methyl ester (L-NAME) l(-1), their development in vitro was arrested at the two cell stage. This inhibition of development was overcome by the addition of 1 mmol L-arginine l(-1) to the medium. These observations indicate that nitric oxide plays an important role as a diffusible regulator of cell proliferation and differentiation, especially at the developmental transition from the two-cell to the four-cell stage during preimplantation development of mice. PMID- 11732992 TI - Expression of oxytocin, oestrogen and progesterone receptors in uterine biopsy samples throughout the oestrous cycle and early pregnancy in cows. AB - This study examined the expression patterns of oxytocin and steroid receptors in the bovine endometrium during the oestrous cycle and early pregnancy to elucidate their respective roles in the regulation of luteolysis and the maternal recognition of pregnancy. In Expt 1, uterine biopsies were collected from four cows throughout three oestrous cycles each, to provide daily samples. In Expt 2, uterine tissue was collected on days 12, 14, 16 and 18 of the oestrous cycle (n = 20) or early pregnancy (n = 16). Oxytocin receptor, oestrogen receptor alpha and progesterone receptor mRNAs were localized by in situ hybridization, and localization of oestrogen receptor and progesterone receptor was confirmed by immunocytochemistry. All three receptors showed time- and cell-specific expression patterns. Oestrogen receptor alpha increased in all regions at oestrus but high concentrations were also found in the luminal epithelium during the mid luteal phase and in the deep glands throughout the oestrous cycle. Progesterone receptor expression was higher in the stroma than it was in the types of epithelial cell, and increased expression was observed at oestrus and during the early luteal phase. The cyclical upregulation of oxytocin receptors in the luminal epithelium on about day 16 was not related to preceding changes in the endometrial expression of either oestradiol alpha or progesterone receptors. During early pregnancy, oxytocin receptor expression was suppressed. Oestrogen receptor a concentrations increased in the non-pregnant cows and decreased in the pregnant cows between days 16 and 18, but these changes followed rather than preceded the upregulation of oxytocin receptors in the non-pregnant cows. It is concluded that the initial upregulation of oxytocin receptors in the luminal epithelium, which triggers luteolysis, is not associated directly with changes in expression of oestrogen receptor alpha. PMID- 11732993 TI - Purification and characterization of three galactose specific lectins from mulberry seeds (Morus sp.). AB - Three lectins were extracted and purified from mulberry seeds by gel filtration of 100% ammonium sulfate saturated crude protein extract followed by ion-exchange chromatography on DEAE and CM-cellulose. The lectins were found to be homogeneous as judged by polyacrylamide disc gel electrophoresis. The molecular masses of the lectins as determined by gel filtration were 175 000 for MSL-1, 120 000 for MSL-2 and 89 500 for MSL-3. MSL-1 is dimer in nature, with the two monomers held together by disulfide bond(s), while MSL-2 and MSL-3 contain four nonidentical subunits that are held together by nonionic hydrophobic interactions. The lectins agglutinated rat red blood cells and this agglutination was inhibited specifically by galactose, methyl-alpha-d-galactopyranoside, methyl-beta-d galactopyranoside, lactose and raffinose. The lectins MSL-1, MSL-2 and MSL-3 contained 5.7, 5.4 and 4.5% neutral sugars, respectively, and the sugar composition of the lectins was glucose and mannose for MSL-1 and galactose for both MSL-2 and MSL-3. The lectins exhibited strong cytotoxic effect in brine shrimp lethality bioassay. PMID- 11732994 TI - Characterization of the active site of histidine ammonia-lyase from Pseudomonas putida. AB - Elucidation of the 3D structure of histidine ammonia-lyase (HAL, EC 4.3.1.3) from Pseudomonas putida by X-ray crystallography revealed that the electrophilic prosthetic group at the active site is 3,5-dihydro-5-methylidene-4H-imidazol-4 one (MIO) [Schwede, T.F., Retey, J., Schulz, G.E. (1999) Biochemistry, 38, 5355 5361]. To evaluate the importance of several amino-acid residues at the active site for substrate binding and catalysis, we mutated the following amino-acid codons in the HAL gene: R283, Y53, Y280, E414, Q277, F329, N195 and H83. Kinetic measurements with the overexpressed mutants showed that all mutations resulted in a decrease of catalytic activity. The mutants R283I, R283K and N195A were approximately 1640, 20 and 1000 times less active, respectively, compared to the single mutant C273A, into which all mutations were introduced. Mutants Y280F, F329A and Q277A exhibited approximately 55, 100 and 125 times lower activity, respectively. The greatest loss of activity shown was in the HAL mutants Y53F, E414Q, H83L and E414A, the last being more than 20 900-fold less active than the single mutant C273A, while H83L was 18 000-fold less active than mutant C273A. We propose that the carboxylate group of E414 plays an important role as a base in catalysis. To investigate a possible participation of active site amino acids in the formation of MIO, we used the chromophore formation upon treatment of HAL with l-cysteine and dioxygen at pH 10.5 as an indicator. All mutants, except F329A showed the formation of a 338-nm chromophore arising from a modified MIO group. The UV difference spectra of HAL mutant F329A with the MIO-free mutant S143A provide evidence for the presence of a MIO group in HAL mutant F329A also. For modelling of the substrate arrangement within the active site and protonation state of MIO, theoretical calculations were performed. PMID- 11732995 TI - Supermolecular organization of photosystem II and its associated light-harvesting antenna in Arabidopsis thaliana. AB - The organization of Arabidopsis thaliana photosystem II (PSII) and its associated light-harvesting antenna (LHCII) was studied in isolated PSII-LHCII supercomplexes and native membrane-bound crystals by transmission electron microscopy and image analysis. Over 4000 single-particle projections of PSII LHCII supercomplexes were analyzed. In comparison to spinach supercomplexes [Boekema, E.J., van Roon, H., van Breemen, J.F.L. & Dekker, J.P. (1999) Eur. J. Biochem. 266, 444-452] some striking differences were revealed: a much larger number of supercomplexes from Arabidopsis contain copies of M-type LHCII trimers. M-type trimers can also bind in the absence of the more common S-type trimers. No binding of l-type trimers could be detected. Analysis of native membrane-bound PSII crystals revealed a novel type of crystal with a unit cell of 25.6 x 21.4 nm (angle 77 degrees ), which is larger than any of the PSII lattices observed before. The data show that the unit cell is built up from C2S2M2 supercomplexes, rather than from C2S2M supercomplexes observed in native membrane crystals from spinach [Boekema, E.J., Van Breemen, J.F.L., Van Roon, H. & Dekker, J.P. (2000) J. Mol. Biol. 301, 1123-1133]. It is concluded from both the single particle analysis and the crystal analysis that the M-type trimers bind more strongly to PSII core complexes in Arabidopsis than in spinach. PMID- 11732996 TI - Calcium-binding by p26olf, an S100-like protein in the frog olfactory epithelium. AB - Frog p26olf is a novel S100-like Ca2+-binding protein found in olfactory cilia. It consists of two S100-like domains aligned sequentially, and has a total of four Ca2+-binding sites (known as EF-hands). In this study, to elucidate the mechanism of Ca2+-binding to each EF-hand (named EF-A, -B, -C and -D from the N terminus of p26olf), we examined Ca2+-binding in wild-type p26olf and also in its mutants in which a glutamate at the -z coordinate position within each Ca2+ binding loop was substituted for a glutamine. Flow dialysis experiments showed that the wild-type binds nearly four Ca2+ per molecule maximally, while all the mutants bind approximately three Ca2+. Although EF-B and -D are p26olf-specific EF-hands and their role in Ca2+-binding is not known, the result unequivocally showed that they actually bind Ca2+. The overall Ca2+-binding affinity decreased in the three mutants. The decrease was very large in the mutants of EF-A and -B, which suggested that the Ca2+-affinities are high in EF-A and -B in the wild type. Assuming the presence of four steps of Ca2+-binding, we determined the dissociation constant of each step in wild-type p26olf. To assign which step takes place at which EF-hand, we measured the antagonistic effect of K+ on each step, as the effect of K+ is thought to be a function of the number of the carboxyl groups in an EF-hand. Although the actual Ca2+-binding mechanism may not be so simple, this study together with the mutation study suggested a tentative Ca2+-binding model of p26olf: the order of Ca2+-binding to p26olf is EF-B, EF-A, EF-C and EF-D. Based on these results, we speculate that similar Ca2+-binding takes place in an S100 dimer. PMID- 11732997 TI - Identification of a critical lysine residue at the active site in glyceraldehyde 3-phosphate dehydrogenase of Ehrlich ascites carcinoma cell. Comparison with the rabbit muscle enzyme. AB - The involvement of the lysine residue present at the active site of Ehrlich ascites carcinoma (EAC) cell glyceraldehyde-3-phosphate dehydrogenase (Gra3PDH) was investigated by using the lysine specific reagents trinitrobenzenesulfonic acid (TNBS) and pyridoxal phosphate (PP). Both TNBS and PP inactivated EAC cell Gra3PDH with pseudo-first-order kinetics with the rate dependent on modifier concentration. Kinetic analysis, including a Tsou plot, indicated that both TNBS and PP apparently react with one lysine residue per enzyme molecule. Two of the substrates, d-glyceraldehyde-3-phosphate and NAD, and also NADH, the product and competitive inhibitor, almost completely protected the enzyme from inactivation by TNBS. A comparative study of Gra3PDH of EAC cell and rabbit muscle indicates that the nature of active site of the enzyme is significantly different in these two cells. A double inhibition study using 5,5'-dithiobis(2-nitrobenzoic acid) and TNBS and subsequent reactivation of only the rabbit muscle enzyme by dithiothreitol suggested that a cysteine residue of this enzyme possibly reacts with TNBS. These studies on the other hand, confirm that an essential lysine residue is involved in the catalytic activity of the EAC cell enzyme. This difference in the nature of the active site of EAC cell Gra3PDH that may be related to the high glycolysis of malignant cells has been discussed. PMID- 11732998 TI - Synthesis, conformational analysis and biological activity of cyclic analogs of the octadecaneuropeptide ODN. Design of a potent endozepine antagonist. AB - The octadecaneuropeptide (ODN; QATVGDVNTDRPGLLDLK) and its C-terminal octapeptide (OP; RPGLLDLK), which exert anxiogenic activity, have been previously shown to increase intracellular calcium concentration ([Ca2+]i) in cultured rat astrocytes through activation of a metabotropic receptor positively coupled to phospholipase C. It has also been found that the [d-Leu5]OP analog possesses a weak antagonistic activity. The aim of the present study was to synthesize and characterize cyclic analogs of OP and [d-Leu5]OP. On-resin homodetic backbone cyclization of OP yielded an analog, cyclo1-8 OP, which was three times more potent and 1.4-times more efficacious than OP to increase [Ca2+]i in cultured rat astrocytes. Cyclo1-8 OP also mimicked the effect of both OP and ODN on polyphosphoinositide turnover. Conversely, the cyclo1-8 [d-Leu5]OP analog was totally devoid of agonistic activity but suppressed the effect of OP and ODN on [Ca2+]i and phosphoinositide metabolism in astrocytes. The structure of these cyclic analogs has been determined by two-dimensional 1H-NMR and molecular dynamics. Cyclo1-8 OP exhibited a single conformation characterized by a gamma turn comprising residues Pro2-Leu4 and a type III beta turn encompassing residues Leu5-Lys8. Cyclo1-8 [d-Leu5]OP was present as two equimolar conformers resulting from cis/trans isomerization of the Arg-Pro peptide bond. These pharmacological and structural data should prove useful for the rational design of non peptidic ODN analogs. PMID- 11732999 TI - CTGF/Hcs24 induces chondrocyte differentiation through a p38 mitogen-activated protein kinase (p38MAPK), and proliferation through a p44/42 MAPK/extracellular signal regulated kinase (ERK). AB - Connective tissue growth factor/hypertrophic chondrocyte specific gene product 24 (CTGF/Hcs24) promotes proliferation and differentiation of chondrocytes in culture. We investigated the roles of two major types of mitogen activated protein kinase (MAPK) in the promotion of proliferation and differentiation by CTGF/Hcs24. Here we report the effects of the MAPKK/MEK 1/2 inhibitor, PD098059, and p38 MAPK inhibitor, SB203580, in a human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) and rabbit growth cartilage (RGC) cells treated with CTGF/Hcs24. In the proliferation phase, CTGF/Hcs24 induced a approximately fivefold increase in the phosphorylation of p44/42 MAPK/ERK and a approximately twofold increase in that of p38 MAPK in an in vivo kinase assay. These inhibitors of MAPKK and MAPK suppressed phosphorylation of ets-like gene-1 (Elk-1) and nuclear activating transcription factor-2 (Atf-2) induced by CTGF/Hcs24 in a dose dependent manner, respectively. Western blot analysis showed that phosphorylation of ERK was induced from 30 to 60 min and phosphorylation of p38 MAPK from 10 to 15 min after the addition of CTGF/Hcs24 in confluence HCS-2/8 cells. PD098059 suppressed the DNA synthesis of HCS-2/8 cells and RGC cells, while SB203580 did not. On the other hand, the p38 MAPK inhibitor, SB203580, completely inhibited the CTGF/Hcs24-induced synthesis of proteoglycans in HCS-2/8 cells and RGC cells but the MEK1/2 inhibitor, PD098059, did not. These results suggest that ERK mediates the CTGF/Hcs24-induced proliferation of chondrocytes, and that p38 MAPK mediates the CTGF/Hcs24-induced differentiation of chondrocytes. PMID- 11733000 TI - Molecular interactions with nuclear factor kappaB (NF-kappaB) transcription factors of a PNA-DNA chimera mimicking NF-kappaB binding sites. AB - The decoy approach against nuclear factor kappaB (NF-kappaB) is a useful tool to alter NF-kappaB dependent gene expression using synthetic oligonucleotides (ODNs) carrying NF-kappaB specific cis-elements. Unfortunately, ODNs are not stable and need to be be extensively modified to be used in vivo or ex vivo. We have previously evaluated the possible use of peptide nucleic acids (PNAs) as decoy molecules. The backbone of PNAs is composed of N-(2-aminoethyl)glycine units, rendering these molecules resistant to both nucleases and proteases. We found that the binding of NF-kappaB transcription factors to PNAs was either very low (binding to PNA-PNA hybrids) or exhibited low stability (binding to PNA-DNA hybrids). The main consideration of the present paper was to determine whether PNA-DNA chimeras mimicking NF-kappaB binding sites are capable of stable interactions with proteins belonging to the NF-kappaB family. Molecular modeling was employed for the design of PNA-DNA chimeras; prediction of molecular interactions between chimeras and NF-kappaB nuclear proteins were investigated by molecular dynamics simulations, and interactions between PNA-DNA chimeras and NF kappaB proteins were studied by gel shifts. We found significant differences between the structure of duplex NF-kappaB PNA-DNA chimera and duplex NF-kappaB DNA-DNA. However, it was found that these differences do not prevent the duplex PNA-DNA chimera from binding to NF-kappaB transcription factors, being able to suppress the molecular interactions between HIV-1 LTR and p50, p52 and nuclear factors from B-lymphoid cells. Therefore, these results demonstrate that the designed NF-kappaB DNA-PNA chimeras could be used for a decoy approach in gene therapy. PMID- 11733001 TI - ik3-1/Cables is a substrate for cyclin-dependent kinase 3 (cdk 3). AB - p70ik3-1 (a 70-kDa protein) contains a cyclin box, and binds to p35cdk3 in vivo and in vitro [Matsuoka, M., Matsuura, Y., Semba, K. & Nishimoto, I. (2000) Biochem. Biophys. Res. Commun. 273, 442-447]. In spite of its structural similarity to cyclins, p70ik3-1 does not activate cyclin-dependent kinase 3 (cdk3)-mediated phosphorylation of pRb, histone H1, or the C-terminal domain of RNA polymerase II. Here, we report that Ser274 of p70ik3-1 is phosphorylated by cdk2 or cdk3 bound to cyclin A and to cyclin E in vitro. We also found that in COS7 cells in which cyclin E and cdk3 were ectopically overexpressed, the phosphorylation level of Ser274 in coexpressed p70ik3-1 is upregulated. We therefore conclude that p70ik3-1 is a substrate for cdk3-mediated phosphorylation. PMID- 11733002 TI - A new siglec family member, siglec-10, is expressed in cells of the immune system and has signaling properties similar to CD33. AB - The siglecs (sialic acid-binding Ig-like lectins) are a distinct subset of the Ig superfamily with adhesion-molecule-like structure. We describe here a novel member of the siglec protein family that shares a similar structure including five Ig-like domains, a transmembrane domain, and a cytoplasmic tail containing two ITIM-signaling motifs. Siglec-10 was identified through database mining of an asthmatic eosinophil EST library. Using the Stanford G3 radiation hybrid panel we were able to localize the genomic sequence of siglec-10 within the cluster of genes on chromosome 19q13.3-4 that encode other siglec family members. We have demonstrated that siglec-10 is an immune system-restricted membrane-bound protein that is highly expressed in peripheral blood leukocytes as demonstrated by Northern, RT-PCR and flow cytometry. Binding assays determined that the extracellular domain of siglec-10 was capable of binding to peripheral blood leukocytes. The cytoplasmic tail of siglec-10 contains four tyrosines, two of which are embedded in ITIM-signaling motifs (Y597 and Y667) and are likely involved in intracellular signaling. The ability of tyrosine kinases to phosphorylate the cytoplasmic tyrosines was evaluated by kinase assay using wild type siglec-10 cytoplasmic domain and Y-->F mutants. The majority of the phosphorylation could be attributed to Y597 andY667. Further experiments with cell extracts suggest that SHP-1 interacts with Y667 and SHP-2 interacts with Y667 in addition to another tyrosine. This is very similar to CD33, which also binds the phosphatases SHP-1 and SHP-2, therefore siglec-10, as CD33, may be characterized as an inhibitory receptor. PMID- 11733003 TI - Expression of V-ATPase proteolipid subunit of Acetabularia acetabulum in a VMA3 deficient strain of Saccharomyces cerevisiae and its complementation study. AB - The function of the translation products of six different cDNAs for Acetabularia V-ATPase proteolipid subunit (AACEVAPD1 to AACEVAPD6) was examined using a Saccharomyces cerevisiae VMA3-deficient strain that lacked its own gene for one of the proteolipid subunits of V-ATPase. Expression of the cDNAs in the strain revealed that four cDNAs from the six complemented the proton transport activity into the vacuole, visualized by fluorescence microscopy. The vacuolar-membrane enriched fractions from the four transformants showed cross-reactivity with antibodies against the subunits a and A of S. cerevisiae V-ATPase. Two translation products from the other two cDNAs were demonstrated not to be localized in vacuolar membranes, and thus could not complement the function of the VMA3-deficient strain. As the primary structures deduced from the former four cDNAs are similar but clearly different from those of the latter two, the latter two translation products may not be able to substitute for theVMA3 gene product. PMID- 11733004 TI - Immunohistochemical localization of guinea-pig leukotriene B4 12 hydroxydehydrogenase/15-ketoprostaglandin 13-reductase. AB - We have cloned cDNA for leukotriene B4 12-hydroxydehydrogenase (LTB4 12-HD)/15 ketoprostaglandin 13-reductase (PGR) from guinea-pig liver. LTB4 12-HD catalyzes the conversion of LTB4 into 12-keto-LTB4 in the presence of NADP+, and plays an important role in inactivating LTB4. The cDNA contained an ORF of 987 bp that encodes a protein of 329 amino-acid residues with a 78% identity with porcine LTB4 12-HD. The amino acids in the putative NAD+/NADP+ binding domain are well conserved among the pig, guinea-pig, human, rat, and rabbit enzymes. The guinea pig LTB4 12-HD (gpLTB4 12-HD) was expressed as a glutathione S-transferase (GST) fusion protein in Escherichia coli, which exhibited similar enzyme activities to porcine LTB4 12-HD. We examined the 15-ketoprostaglandin 13-reductase (PGR) activity of recombinant gpLTB4 12-HD, and confirmed that the Kcat of the PGR activity is higher than that of LTB4 12-HD activity by 200-fold. Northern and Western blot analyses revealed that gpLTB4 12-HD/PGR is widely expressed in guinea-pig tissues such as liver, kidney, small intestine, spleen, and stomach. We carried out immunohistochemical analyses of this enzyme in various guinea-pig tissues. Epithelial cells of calyx and collecting tubules in kidney, epithelial cells of airway, alveoli, epithelial cells in small intestine and stomach, and hepatocytes were found to express the enzyme. These findings will lead to the identification of the unrevealed roles of PGs and LTs in these tissues. PMID- 11733005 TI - Production and chemiluminescent free radical reactions of glyoxal in lipid peroxidation of linoleic acid by the ligninolytic enzyme, manganese peroxidase. AB - Glyoxal is a key compound involved in glyoxal oxidase (GLOX)-dependent production of glyoxylate, oxalate and H2O2 by lignin-degrading basidiomycetes. In this paper, we report that glyoxal was produced from a metabolite of ligninolytic fungi, linoleic acid, by manganese peroxidase (MnP)-dependent lipid peroxidation. In the absence of the parent substrate of linoleic acid, the dialdehyde was oxidized by MnP and Mn(III) chelate to start free radical reactions with emission of chemiluminescence at 700-710 nm. The spectroscopic profile of the light emission is distinguishable from (a) singlet oxygen, (b) triplet carbonyls from dioxetane and alpha-hydroxyperoxyl radicals, and (c) biacyl triplet formed by the coupling of two acyl radicals. The photon emission of glyoxal by MnP was activated by co-oxidation of tartrate. The MnP-dependent oxidation of glyoxal in tartrate buffers continued for 10 days without addition of exogenous H2O2. The importance of these results is discussed in relation to the free radical chemistry of lignin biodegradation by wood rot fungi. PMID- 11733006 TI - Isolation, enzymatic properties, and mode of action of an exo-1,3-beta-glucanase from Trichoderma viride. AB - An exo-1,3-beta-glucanase has been isolated from cultural filtrate of T. viride AZ36. The N-terminal sequence of the purified enzyme (m = 61 +/- 1 kDa) showed no significant homology to other known glucanases. The 1,3-beta-glucanase displayed high activity against laminarins, curdlan, and 1,3-beta-oligoglucosides, but acted slowly on 1,3-1,4-beta-oligoglucosides. No significant activity was detected against high molecular mass 1,3-1,4-beta-glucans. The enzyme carried out hydrolysis with inversion of the anomeric configuration. Whereas only glucose was released from the nonreducing terminus during hydrolysis of 1,3-beta oligoglucosides, transient accumulation of gentiobiose was observed during hydrolysis of laminarins. The gentiobiose was subsequently degraded to glucose. The Michaelis constants Km and Vmax have been determined for the hydrolysis of 1,3-beta-oligoglucosides with degrees of polymerization ranging from 2 to 6. Based on these data, binding affinities for subsites were calculated. Substrate binding site contained at least five binding sites for sugar residues. PMID- 11733007 TI - Ontogeny and subcellular localization of rat liver mitochondrial branched chain amino-acid aminotransferase. AB - Branched chain amino-acid aminotransferase (BCAT) activity is present in fetal liver but the developmental pattern of mitochondrial BCAT (BCATm) expression in rat liver has not been studied. The aim of this study was to determine the activity, protein and mRNA concentration of BCATm in fetal and postnatal rat liver, and to localize this enzyme at the cellular and subcellular levels at both developmental stages. Maximal BCAT activity and BCATm mRNA expression occurred at 17 days' gestation in fetal rat liver and then declined significantly immediately after birth. This pattern was observed only in liver; rat heart showed a different developmental pattern. Fetal liver showed intense immunostaining to BCATm in the nuclei and mitochondria of hepatic cells and blood cell precursors; in contrast, adult liver showed mild immunoreactivity located only in the mitochondria of hepatocytes. BCAT activity in isolated fetal liver nuclei was 0.64 mU x mg(-1) protein whereas it was undetectable in adult liver nuclei. By Western blot analysis the BCATm antibody recognized a 41-kDa protein in fetal liver nuclei, and proteins of 41 and 43 kDa in fetal liver supernatant. In adult rat liver supernatant, the BCATm antibody recognized only a 43-kDa protein; however, neither protein was detected in adult rat liver nuclei. The appearance of the 41-kDa protein was associated with the presence of the highly active form of BCATm. These results suggest the existence of active and inactive forms of BCAT in rat liver. PMID- 11733008 TI - Comparison of starch branching enzyme I and II from potato. AB - The in vitro activities of purified potato starch branching enzyme (SBE) I and II expressed in Escherichia coli were compared using several assay methods. With the starch-iodine method, it was found that SBE I was more active than SBE II on an amylose substrate, whereas SBE II was more active than SBE I on an amylopectin substrate. Both enzymes were stimulated by the presence of phosphate. On a substrate consisting of linear dextrins (chain length 8-200 glucose residues), no significant net increase in molecular mass was seen on gel-permeation chromatography after incubation with the enzymes. This indicates intrachain branching of the substrate. After debranching of the products, the majority of dextrins with a degree of polymerization (dp) greater than 60 were absent for SBE I and those with a dp greater than 70 for SBE II. To study the shorter chains, the debranched samples were also analysed by high-performance anion-exchange chromatography. The products of SBE I showed distinct populations at dp 11-12 and dp 29-30, whereas SBE II products had one, broader, population with a peak at dp 13-14. An accumulation of dp 6-7 chains was seen with both isoforms. PMID- 11733009 TI - A neuropeptide Y receptor Y1-subfamily gene from an agnathan, the European river lamprey. A potential ancestral gene. AB - We report here the isolation and functional expression of a neuropeptide Y (NPY) receptor from the river lamprey, Lampetra fluviatilis. The receptor displays approximately 50% amino-acid sequence identity to all previously cloned Y1 subfamily receptors including Y1, Y4, and y6 and the teleost subtypes Ya, Yb and Yc. Phylogenetic analyses point to a closer relationship with Y4 and Ya/b/c suggesting that the lamprey receptor could possibly represent a pro-orthologue of some or all of those gnathostome receptors. Our results support the notion that the Y1 subfamily increased in number by genome or large-scale chromosome duplications, one of which may have taken place prior to the divergence of lampreys and gnathostomes whereas the second duplication probably occurred in the gnathostome lineage after this split. Functional expression of the lamprey receptor in a cell line facilitated specific binding of the three endogenous lamprey peptides NPY, peptide YY and peptide MY with picomolar affinities. Binding studies with a large panel of NPY analogues revealed indiscriminate binding properties similar to those of another nonselective Y1-subfamily receptor, zebrafish Ya. RT-PCR detected receptor mRNA in the central nervous system as well as in several peripheral organs suggesting diverse functions. This lamprey receptor is evolutionarily the most distant NPY receptor that clearly belongs to the Y1 subfamily as defined in mammals, which shows that subtypes Y2 and Y5 arose even earlier in evolution. PMID- 11733010 TI - Toxicity of novel C-terminal prion protein fragments and peptides harbouring disease-related C-terminal mutations. AB - Mice expressing a C-terminal fragment of the prion protein instead of wild-type prion protein die from massive neuronal degeneration within weeks of birth. The C terminal region of PrPc (PrP121-231) expressed in these mice has an intrinsic neurotoxicity to cultured neurones. Unlike PrPSc, which is not neurotoxic to neurones lacking PrPc expression, PrP121-231 was more neurotoxic to PrPc deficient cells. Human mutations E200K and F198S were found to enhance toxicity of PrP121-231 to PrP-knockout neurones and E200K enhanced toxicity to wild-type neurones. The normal metabolic cleavage point of PrPc is approximately amino-acid residue 113. A fragment of PrPc corresponding to the whole C-terminus of PrPc (PrP113-231), which is eight amino acids longer than PrP121-231, lacked any toxicity. This suggests the first eight amino residues of PrP113-121 suppress toxicity of the toxic domain in PrP121-231. Addition to cultures of a peptide (PrP112-125) corresponding to this region, in parallel with PrP121-231, suppressed the toxicity of PrP121-231. These results suggest that the prion protein contains two domains that are toxic on their own but which neutralize each other's toxicity in the intact protein. Point mutations in the inherited forms of disease might have their effects by diminishing this inhibition. PMID- 11733011 TI - Binding of gelsolin domain 2 to actin. An actin interface distinct from that of gelsolin domain 1 and from ADF/cofilin. AB - It is generally assumed that of the six domains that comprise gelsolin, domain 2 is primarily responsible for the initial contact with the actin filament that will ultimately result in the filament being severed. Other actin-binding regions within domains 1 and 4 are involved in gelsolin's severing and subsequent capping activity. The overall fold of all gelsolin repeated domains are similar to the actin depolymerizing factor (ADF)/cofilin family of actin-binding proteins and it has been proposed that there is a similarity in the actin-binding interface. Gelsolin domains 1 and 4 bind G-actin in a similar manner and compete with each other, whereas domain 2 binds F-actin at physiological salt concentrations, and does not compete with domain 1. Here we investigate the domain 2 : actin interface and compare this to our recent studies of the cofilin : actin interface. We conclude that important differences exist between the interfaces of actin with gelsolin domains 1 and 2, and with ADF/cofilin. We present a model for F-actin binding of domain 2 with respect to the F-actin severing and capping activity of the whole gelsolin molecule. PMID- 11733012 TI - Identification of an element within the promoter of human selenoprotein P responsive to transforming growth factor-beta. AB - Selenoprotein P (SeP) is a plasma protein that contains up to 10 selenocysteine residues and accounts for about 50% of total selenium in human plasma. We have previously shown that SeP expression in the human liver cell line HepG2 is inhibited by transforming growth factor (TGF)-beta1 on a transcriptional level. Smad proteins are the transcriptional mediators of TGF-beta signalling and putative Smad-binding elements (SBE) comprising the core sequence CAGACA are present at two positions in the SeP promoter. The aim of our study was to investigate whether Smad molecules are involved in inhibition of SeP expression by TGF-beta1 and to locate the promoter region critical for this effect. As seen in electrophoretic-mobility-shift assays, TGF-beta1 treatment led to enhanced binding of nuclear proteins to a putative SBE from the SeP promoter. Overexpression of Smad 3 and 4, but not of Smad 2, resulted in a marked down regulation of SeP mRNA expression. Similar effects were observed for luciferase expression under control of a human SeP-promoter construct. Deletion as well as point-mutation of putative SBEs led to a loss of promoter sensitivity towards TGF beta1 treatment. Hence, we demonstrated an involvement of Smad 3 and 4 in transcriptional regulation of SeP by TGF-beta1 and we were able to identify the TGF-beta-responsive element in the SeP promoter. PMID- 11733013 TI - Progressive apoptosis in chorion laeve trophoblast cells of human fetal membrane tissues during in vitro incubation is suppressed by antioxidative reagents. AB - Previously, we demonstrated apoptotic cell death in the chorion laeve trophoblast layer of human fetal membrane tissues during the late stages of pregnancy, the progression of apoptosis during incubation in vitro, and its suppression by a low concentration of glucocorticoid hormones. We now report examination of mRNA expression of inflammatory cytokines [interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha] and antioxidative enzyme genes [heme oxygenase 1, catalase, Mn-superoxide dismutase (SOD), Cu/Zn-SOD, glutathione S-transferase, glutathione reductase and glutathione peroxidase] and apoptosis-related genes during in vitro progression of apoptosis with or without glucocorticoid by a reverse transcription/PCR method. It was shown that the mRNA levels increased in chorion laeve tissue for each cytokine examined and for catalase, heme oxygenase 1 and Mn-SOD in direct correlation with the in vitro incubation period. By Western blotting the existence of Mn-SOD protein, and its slight increase with incubation time, was also shown. The investigation of the influence of antioxidative reagents [pyrrolidine dithiocarbamate (PDTC), N-acetyl-l-cysteine (NAC) and nordihydroguaiaretic acid (NDGA)] on DNA fragmentation showed that DNA fragmentation in chorion laeve tissues was inhibited by approximately 50% in the presence of 1 mm PDTC, 30 mm NAC and 1 mm NDGA. These results suggest that apoptotic cell death of the trophoblast layer of chorion tissues may be induced through intracellular oxidative stress at the stage of parturition. PMID- 11733014 TI - Arginine 121 is a crucial residue for the specific cytotoxic activity of the ribotoxin alpha-sarcin. AB - Alpha-sarcin, a cyclizing ribonuclease secreted by the mould Aspergillus giganteus, is one of the best characterized members of a family of fungal ribotoxins. This protein induces apoptosis in tumour cells due to its highly specific activity on ribosomes. Fungal ribotoxins display a three-dimensional protein fold similar to those of a larger group of microbial noncytotoxic RNases, represented by RNases T1 and U2. This similarity involves the three catalytic residues and also the Arg121 residue, whose counterpart in RNase T1, Arg77, is located in the vicinity of the substrate phosphate moiety although its potential functional role is not known. In this work, Arg121 of alpha-sarcin has been replaced by Gln or Lys. These two mutations do not modify the conformation of the protein but abolish the ribosome-inactivating activity of alpha-sarcin. In addition, the loss of the positive charge at that position produces dramatic changes on the interaction of alpha-sarcin with phospholipid membranes. It is concluded that Arg121 is a crucial residue for the characteristic cytotoxicity of alpha-sarcin and presumably of the other fungal ribotoxins. PMID- 11733015 TI - The regulatory subunit of a cGMP-regulated protein kinase A of Trypanosoma brucei. AB - This study reports the identification and characterization of the regulatory subunit, TbRSU, of protein kinase A of the parasitic protozoon Trypanosoma brucei. TbRSU is coded for by a single copy gene. The protein contains an unusually long N-terminal domain, the pseudosubstrate site involved in binding and inactivation of the catalytic subunit, and two C-terminally located, closely spaced cyclic nucleotide binding domains. Immunoprecipitation of TbRSU coprecipitates a protein kinase activity with the characteristics of protein kinase A: it phosphorylates a protein kinase specific substrate, and it is strongly inhibited by a synthetic protein kinase inhibitor peptide. Unexpectedly, this kinase activity could not be stimulated by cAMP, but by cGMP only. Binding studies with recombinant cyclic nucleotide binding domains of TbRSU confirmed that both domains bind cGMP with Kd values in the lower micromolar range, and that up to a 100-fold excess of cAMP does not compete with cGMP binding. PMID- 11733016 TI - Shifted positioning of the anticodon nucleotide residues of amber suppressor tRNA species by Escherichia coli arginyl-tRNA synthetase. AB - Cytidine in the anticodon second position (position 35) and G or U in position 36 of tRNAArg are required for aminoacylation by arginyl-tRNA synthetase (ArgRS) from Escherichia coli. Nevertheless, an arginine-accepting amber suppressor tRNA with a CUA anticodon (FTOR1Delta26) exhibits suppression activity in vivo [McClain, W.H. & Foss, K. (1988) Science, 241, 1804-1807]. By an in vitro kinetic study with mutagenized tRNAs, we showed that the arginylation of FTOR1Delta26 involves C34 and U35, and that U35 can be replaced by G without affecting the activity. Thus, the positioning of the essential nucleotides for the arginylation is shifted to the 5' side, by one residue, in the suppressor tRNAArg. We found that the shifted positioning does not depend on the tRNA sequence outside the anticodon. Furthermore, by a genetic method, we isolated a mutant ArgRS that aminoacylates FTOR1Delta26 more efficiently than the wild-type ArgRS. The isolated mutant has mutations at two nonsurface amino-acid residues that interact with each other near the anticodon-binding site. PMID- 11733017 TI - Effect of ibuprofen and warfarin on the allosteric properties of haem-human serum albumin. A spectroscopic study. AB - Haem binding to human serum albumin (HSA) endows the protein with peculiar spectroscopic properties. Here, the effect of ibuprofen and warfarin on the spectroscopic properties of ferric haem-human serum albumin (ferric HSA-haem) and of ferrous nitrosylated haem-human serum albumin (ferrous HSA-haem-NO) is reported. Ferric HSA-haem is hexa-coordinated, the haem-iron atom being bonded to His105 and Tyr148. Upon drug binding to the warfarin primary site, the displacement of water molecules--buried in close proximity to the haem binding pocket--induces perturbation of the electronic absorbance properties of the chromophore without affecting the coordination number or the spin state of the haem-iron, and the quenching of the 1H-NMR relaxivity. Values of Kd for ibuprofen and warfarin binding to the warfarin primary site of ferric HSA-haem, corresponding to the ibuprofen secondary cleft, are 5.4 +/- 1.1 x 10(-4) m and 2.1 +/- 0.4 x 10(-5) m, respectively. The affinity of ibuprofen and warfarin for the warfarin primary cleft of ferric HSA-haem is lower than that reported for drug binding to haem-free HSA. Accordingly, the Kd value for haem binding to HSA increases from 1.3 +/- 0.2 x 10(-8) m in the absence of drugs to 1.5 +/- 0.2 x 10(-7) m in the presence of ibuprofen and warfarin. Ferrous HSA-haem-NO is a five coordinated haem-iron system. Drug binding to the warfarin primary site of ferrous HSA-haem-NO induces the transition towards the six-coordinated haem-iron species, the haem-iron atom being bonded to His105. Remarkably, the ibuprofen primary cleft appears to be functionally and spectroscopically uncoupled from the haem site of HSA. Present results represent a clear-cut evidence for the drug induced shift of allosteric equilibrium(a) of HSA. PMID- 11733018 TI - Inhibition of glycosyl-phosphatidylinositol biosynthesis in Plasmodium falciparum by C-2 substituted mannose analogues. AB - Mannose analogues (2-deoxy-D-glucose, 2-deoxy-2-fluoro-D-glucose and 2-amino-2 deoxy-D-mannose) have been used to study glycosylphosphatidylinositol (GPtdIns) biosynthesis and GPtdIns protein anchoring in protozoal and mammalian systems. The effects of these analogues on GPtdIns biosynthesis and GPtdIns-protein anchoring of the human malaria parasite Plasmodium falciparum were evaluated in this study. At lower concentrations of 2-deoxy-D-glucose and 2-deoxy-2-fluoro-D glucose (0.2 and 0.1 mm, respectively), GPtdIns biosynthesis is inhibited without significant effects on total protein biosynthesis. At higher concentrations of 2 deoxy-D-glucose and 2-deoxy-2-fluoro-D-glucose (1.5 and 0.8 mm, respectively), the incorporation of [3H]glucosamine into glycolipids was inhibited by 90%, and the attachment of GPtdIns anchor to merozoite surface protein-1 (MSP-1) was prevented. However, at these concentrations, both sugar analogues inhibit MSP-1 synthesis and total protein biosynthesis. In contrast to 2-deoxy-2-fluoro-D glucose and 2-amino-2-deoxy-D-mannose (mannosamine), the formation of new glycolipids was observed only in the presence of tritiated or nonradiolabelled 2 deoxy-D-glucose. Mannosamine inhibits GPtdIns biosynthesis at a concentration of 5 mm, but neither an accumulation of aberrant intermediates nor significant inhibition of total protein biosynthesis was observed in the presence of this analogue. Furthermore, the [3H]mannosamine-labelled glycolipid spectrum resembled the one described for [3H]glucosamine labelling. Total hydrolysis of mannosamine labelled glycolipids showed that half of the tritiated mannosamine incorporated into glycolipids was converted to glucosamine. This high rate of conversion led us to suggest that no actual inhibition from GPtdIns biosynthesis is achieved with the treatment with mannosamine, which is different to what has been observed for mammalian cells and other parasitic protozoa. PMID- 11733019 TI - Structural and biochemical characterization of neuronal calretinin domain I-II (residues 1-100). Comparison to homologous calbindin D28k domain I-II (residues 1 93). AB - This study characterizes the calcium-bound CR I-II domain (residues 1-100) of rat calretinin (CR). CR, with six EF-hand motifs, is believed to function as a neuronal intracellular calcium-buffer and/or calcium-sensor. The secondary structure of CR I-II, defined by standard NMR methods on 13C,15N-labeled protein, contains four helices and two short interacting segments of extended structure between the calcium-binding loops. The linker between the two helix-loop-helix, EF-hand motifs is 12 residues long. Limited trypsinolysis at K60 (there are 10 other K/R residues in CR I-II) confirms that the linker of CR I-II is solvent exposed and that other potential sites are protected by regular secondary structure. 45Ca-overlay of glutathione S-transferase (GST)-CR(1-60) and GST-CR(61 100) fusion proteins confirm that both EF-hands of CR I-II have intrinsic calcium binding properties. The primary sequence and NMR chemical shifts, including calcium-sensitive glycine residues, also suggest that both EF-hand loops of CR I II bind calcium. NMR relaxation, analytical ultracentrifugation, chemical cross linking and NMR translation diffusion measurements indicate that CR I-II exists as a monomer. Calb I-II (the homologous domain of calbindin D28k) has the same EF hand secondary structures as CR I-II, except that helix B is three residues longer and the linker has only four residues [Klaus, W., Grzesiek, S., Labhardt, A. M., Buckwald, P., Hunziker, W., Gross, M. D. & Kallick, D. A. (1999) Eur. J. Biochem. 262, 933-938]. In contrast, Calb I-II binds one calcium cation per monomeric unit and exists as a dimer. Despite close homology and similar secondary structures, CR I-II and Calb I-II probably have distinct tertiary structure features that suggest different cellular functions for the full-length proteins. PMID- 11733020 TI - Structural determinants of the half-life and cleavage site preference in the autolytic inactivation of chymotrypsin. AB - The molecular mechanism of the autolysis of rat alpha-chymotrypsin B was investigated. In addition to the two already known autolytic sites, Tyr146 and Asn147, a new site formed by Phe114 was identified. The former two sites and the latter one are located in the autolysis and the interdomain loops, respectively. By eliminating these sites by site-directed mutagenesis, their involvement in the autolysis and autolytic inactivation processes was studied. Mutants Phe114-->Ile and Tyr146-->His/Asn147-->Ser, that had the same enzymatic activity and molecular stability as the wild-type enzyme, displayed altered routes of autolytic degradation. The Phe114-->Ile mutant also exhibited a significantly slower autolytic inactivation (its half-life was 27-fold longer in the absence and sixfold longer in the presence of Ca2+ ions) that obeyed a first order kinetics instead of the second order displayed by wild-type chymotrypsin inactivation. The comparison of autolysis and autolytic inactivation data showed that: (a) the preferential cleavage of sites followed the order of Tyr146-Asn147 --> Phe114 --> other sites; (b) the cleavage rates at sites Phe114 and Tyr146-Asn147 were independent from each other; and (c) the hydrolysis of the Phe114-Ser115 bond was the rate determining step in autolytic inactivation. Thus, it is the cleavage of the interdomain loop and not of the autolysis or other loops that determines the half-life of chymotrypsin activity. PMID- 11733021 TI - Solution structure of the mEGF/TGFalpha44-50 chimeric growth factor. AB - The solution structure of the growth factor chimera mEGF/TGFalpha44-50 has been determined using an extended version of the dyana procedure for calculating structures from NMR data. The backbone fold and preferred orientation of the domains of the chimera are similar to those found in previous studies of EGF structures, and several H-bonds used as input constraints in those studies were found independently in the chimera. This shows that the modified activity of the chimera does not result from a major structural change. However, the improved precision of the structure presented here allows the origin of some unusual chemical shifts found in all of these compounds to be explained, as well as the results obtained from some site-specific mutants. Further studies of the properties of this chimeric growth factor should help to elucidate the mechanism(s) of hetero- and homodimerization of the c-erbB receptors. PMID- 11733022 TI - Use of site-specific recombination as a probe of nucleoprotein complex formation in chromatin. AB - DNA transactions in eukaryotes require that proteins gain access to target sequences packaged in chromatin. Further, interactions between distinct nucleoprotein complexes are often required to generate higher-order structures. Here, we employed two prokaryotic site-specific recombination systems to investigate how chromatin packaging affects the assembly of nucleoprotein structures of different complexities at more than 30 genomic loci. The dynamic nature of chromatin permitted protein-DNA and DNA-DNA interactions for sites of at least 34 bp in length. However, the assembly of higher-order nucleoprotein structures on targets spanning 114 bp was impaired. This impediment was maintained over at least 72 h and was not affected by the transcriptional status of chromatin nor by inhibitors of histone deacetylases and topoisomerases. Our findings suggest that nucleosomal linker-sized DNA segments become accessible within hours for protein binding due to the dynamic nature of chromatin. Longer segments, however, appear refractory for complete occupancy by sequence-specific DNA-binding proteins. The results thus also provide an explanation why simple recombination systems such as Cre and Flp are proficient in eukaryotic chromatin. PMID- 11733023 TI - Purification, characterization, immunolocalization and structural analysis of the abundant cytoplasmic beta-amylase from Calystegia sepium (hedge bindweed) rhizomes. AB - An abundant catalytically active beta-amylase (EC 3.2.1.2) was isolated from resting rhizomes of hedge bindweed (Calystegia sepium). Biochemical analysis of the purified protein, molecular modeling, and cloning of the corresponding gene indicated that this enzyme resembles previously characterized plant beta-amylases with regard to its amino-acid sequence, molecular structure and catalytic activities. Immunolocalization demonstrated that the beta-amylase is exclusively located in the cytoplasm. It is suggested that the hedge bindweed rhizome beta amylase is a cytoplasmic vegetative storage protein. PMID- 11733024 TI - Importance of the amino-acid composition of the shutter region of plasminogen activator inhibitor-1 for its transitions to latent and substrate forms. AB - The serpins are of general protein chemical interest due to their ability to undergo a large conformational change consisting of the insertion of the reactive centre loop (RCL), which becomes strand 4, into the central beta sheet A. To make space for the incoming RCL, the 'shutter region' opens by the beta strands 3A and 5A sliding apart over the underlying alpha helix B. Loop insertion occurs during the formation of complexes of serpins with their target serine proteinases and during latency transition. This type of loop insertion is unique to plasminogen activator inhibitor-1 (PAI-1). We report here that amino-acid substitutions in a buried cluster of three residues forming a hydrogen bonding network in the shutter region drastically accelerate PAI-1 latency transition; that the rate was in all cases normalized by the PAI-1 binding protein vitronectin; and that substitution of an adjacent beta strand 5A Lys residue, believed to anchor beta strand 5A to other secondary structural elements, had differential effects on the rates of latency transition in the absence and the presence of vitronectin, respectively. An overlapping, but not identical set of substitutions resulted in an increased tendency to substrate behaviour of PAI-1 at reaction with its target proteinases. These findings show that vitronectin regulates the movements of the RCL through conformational changes of the shutter region and beta strand 5A, are in agreement with RCL insertion proceeding by different routes during latency transition and complex formation, and contribute to the biochemical basis for the potential use of PAI-1 as a therapeutic target in cancer and cardiovascular diseases. PMID- 11733025 TI - Coordination structures of Ca2+ and Mg2+ in Akazara scallop troponin C in solution. FTIR spectroscopy of side-chain COO- groups. AB - FTIR spectroscopy has been applied to study the coordination structures of Mg2+ and Ca2+ ions bound in Akazara scallop troponin C (TnC), which contains only a single Ca2+ binding site. The region of the COO- antisymmetric stretch provides information about the coordination modes of COO- groups to the metal ions: bidentate, unidentate, or pseudo-bridging. Two bands were observed at 1584 and 1567 cm-1 in the apo state, whereas additional bands were observed at 1543 and 1601 cm-1 in the Ca2+-bound and Mg2+-bound states, respectively. The intensity of the band at 1567 cm-1 in the Mg2+-bound state was identical to that in the apo state. Therefore, the side-chain COO- group of Glu142 at the 12th position in the Ca2+-binding site coordinates to Ca2+ in the bidentate mode but does not interact with Mg2+ directly. A slight upshift of COO- antisymmetric stretch due to Asp side-chains was also observed upon Mg2+ and Ca2+ binding. This indicates that the COO- groups of Asp131 and Asp133 interact with both Ca2+ and Mg2+ in the pseudo bridging mode. Therefore, the present study directly demonstrated that the coordination structure of Mg2+ was different from that of Ca2+ in the Ca2+ binding site. In contrast to vertebrate TnC, most of the secondary structures remained unchanged among apo, Mg2+-bound and Ca2+-bound states of Akazara scallop TnC, as spectral changes upon either Ca2+ or Mg2+ binding were very small in the infrared amide-I' region as well as in the CD spectra. Fluorescence spectroscopy indicated that the spectral changes upon Ca2+ binding were larger than that upon Mg2+ binding. Moreover, gel-filtration experiments indicated that the molecular sizes of TnC had the order apo TnC > Mg2+-bound TnC > Ca2+-bound TnC. These results suggest that the tertiary structures are different in the Ca2+- and Mg2+ bound states. The present study may provide direct evidence that the side-chain COO- groups in the Ca2+-binding site are directly involved in the functional on/off mechanism of the activation of Akazara scallop TnC. PMID- 11733026 TI - Bivalent cations and amino-acid composition contribute to the thermostability of Bacillus licheniformis xylose isomerase. AB - Comparative analysis of genome sequence data from mesophilic and hyperthermophilic micro-organisms has revealed a strong bias against specific thermolabile amino-acid residues (i.e. N and Q) in hyperthermophilic proteins. The N + Q content of class II xylose isomerases (XIs) from mesophiles, moderate thermophiles, and hyperthermophiles was examined. It was found to correlate inversely with the growth temperature of the source organism in all cases examined, except for the previously uncharacterized XI from Bacillus licheniformis DSM13 (BLXI), which had an N + Q content comparable to that of homologs from much more thermophilic sources. To determine whether BLXI behaves as a thermostable enzyme, it was expressed in Escherichia coli, and the thermostability and activity properties of the recombinant enzyme were studied. Indeed, it was optimally active at 70-72 degrees C, which is significantly higher than the optimal growth temperature (37 degrees C) of B. licheniformis. The kinetic properties of BLXI, determined at 60 degrees C with glucose and xylose as substrates, were comparable to those of other class II XIs. The stability of BLXI was dependent on the metallic cation present in its two metal-binding sites. The enzyme thermostability increased in the order apoenzyme < Mg2+-enzyme < Co2+ enzyme approximately Mn2+-enzyme, with melting temperatures of 50.3 degrees C, 53.3 degrees C, 73.4 degrees C, and 73.6 degrees C. BLXI inactivation was first order in all conditions examined. The energy of activation for irreversible inactivation was also strongly influenced by the metal present, ranging from 342 kJ x mol(-1) (apoenzyme) to 604 kJ x mol(-1) (Mg2+-enzyme) to 1166 kJ x mol(-1) (Co2+-enzyme). These results suggest that the first irreversible event in BLXI unfolding is the release of one or both of its metals from the active site. Although N + Q content was an indicator of thermostability for class II XIs, this pattern may not hold for other sets of homologous enzymes. In fact, the extremely thermostable alpha-amylase from B. licheniformis was found to have an average N + Q content compared with homologous enzymes from a variety of mesophilic and thermophilic sources. Thus, it would appear that protein thermostability is a function of more complex molecular determinants than amino-acid content alone. PMID- 11733027 TI - Studies on the nonmevalonate pathway of terpene biosynthesis. The role of 2C methyl-D-erythritol 2,4-cyclodiphosphate in plants. AB - 2C-methyl-D-erythritol 2,4-cyclodiphosphate was recently shown to be formed from 2C-methyl-D-erythritol 4-phosphate by the consecutive action of IspD, IspE, and IspF proteins in the nonmevalonate pathway of terpenoid biosynthesis. To complement previous work with radiolabelled precursors, we have now demonstrated that [U-13C5]2C-methyl-D-erythritol 4-phosphate affords [U-13C5]2C-methyl-D erythritol 2,4-cyclodiphosphate in isolated chromoplasts of Capsicum annuum and Narcissus pseudonarcissus. Moreover, chromoplasts are shown to efficiently convert 2C-methyl-D-erythritol 4-phosphate as well as 2C-methyl-D-erythritol 2,4 cyclodiphosphate into the carotene precursor phytoene. The bulk of the kinetic data collected in competition experiments with radiolabeled substrates is consistent with the notion that the cyclodiphosphate is an obligatory intermediate in the nonmevalonate pathway to terpenes. Studies with [2,2'-13C2]2C methyl-D-erythritol 2,4-cyclodiphosphate afforded phytoene characterized by pairs of jointly transferred 13C atoms in the positions 17/1, 18/5, 19/9, and 20/13 and, at a lower abundance, in positions 16/1, 4/5, 8/9, and 12/13. A detailed scheme is presented for correlating the observed partial scrambling of label with the known lack of fidelity of the isopentenyl diphosphate/dimethylethyl diphosphate isomerase. PMID- 11733028 TI - Molecular cloning and characterization of isomultiflorenol synthase, a new triterpene synthase from Luffa cylindrica, involved in biosynthesis of bryonolic acid. AB - An oxidosqualene cyclase cDNA, LcIMS1, was isolated from cultured cells of Luffa cylindrica Roem. by heterologous hybridization with cDNA of Glycyrrhiza glabra beta-amyrin synthase. Expression of LcIMS1 in yeast lacking endogenous oxidosqualene cyclase activity resulted in the accumulation of isomultiflorenol, a triterpene. This is consistent with LcIMS1 encoding isomultiflorenol synthase, an oxidosqualene cyclase involved in bryonolic acid biosynthesis in cultured Luffa cells. The deduced amino-acid sequence of LcIMS1 shows relatively low identity with other triterpene synthases, suggesting that isomultiflorenol synthase should be classified into a new group of triterpene synthases. The levels of isomultiflorenol synthase and cycloartenol synthase mRNAs, which were measured with gene-specific probes, correlated with the accumulation of bryonolic acid and phytosterols over a growth cycle of the Luffa cell cultures. Isomultiflorenol synthase mRNA was low during the early stages of cell growth and accumulated to relatively high levels in the late stages. Induction of this mRNA preceded accumulation of bryonolic acid. In contrast, cycloartenol synthase mRNA accumulated in the early stages of the culture cycle, whereas phytosterols accumulated at the same relative rate throughout the whole growth cycle. These results suggest independent regulation of these two genes and of the accumulation of bryonolic acid and phytosterols. PMID- 11733029 TI - Inhibition of the SERCA Ca2+ pumps by curcumin. Curcumin putatively stabilizes the interaction between the nucleotide-binding and phosphorylation domains in the absence of ATP. AB - Curcumin is a compound derived from the spice, tumeric. It is a potent inhibitor of the SERCA Ca2+ pumps (all isoforms), inhibiting Ca2+-dependent ATPase activity with IC50 values of between 7 and 15 microm. It also inhibits ATP-dependent Ca2+ uptake in a variety of microsomal membranes, although for cerebellar and platelet microsomes, a stimulation in Ca2+ uptake is observed at low curcumin concentrations (<10 microm). For the skeletal muscle isoform of the Ca2+ pump (SERCA1), the inhibition of curcumin is noncompetitive with respect to Ca2+, and competitive with respect to ATP at high curcumin concentrations ( approximately 10-25 microm). This was confirmed by ATP binding studies that showed inhibition in the presence of curcumin: ATP-dependent phosphorylation was also reduced. Experiments with fluorescein 5'-isothiocyanate (FITC)-labelled ATPase also suggest that curcumin stabilizes the E1 conformational state. The fact that FITC labels the nucleotide binding site of the ATPase (precluding ATP from binding), and the fact that curcumin affects FITC fluorescence indicate that curcumin must be binding to another site within the ATPase that induces a conformational change to prevent ATP from binding. This observation is interpreted, with the aid of recent structural information, as curcumin stabilizing the interaction between the nucleotide-binding and phosphorylation domains, precluding ATP binding. PMID- 11733030 TI - Roles of nodal-lefty regulatory loops in embryonic patterning of vertebrates. AB - Nodal is a signalling molecule that belongs to the transforming growth factor beta superfamily of proteins, and Lefty proteins are antagonists of Nodal signalling. The nodal and lefty genes form positive and negative regulatory loops that resemble the reaction-diffusion system. As a pair, these genes control various events of vertebrate embryonic patterning, including left-right specification and mesoderm formation. In this review, we will focus on recent studies that have addressed the roles of nodal and lefty in mouse development. PMID- 11733031 TI - Cell culture analysis of the regulatory frameshift event required for the expression of mammalian antizymes. AB - BACKGROUND: Antizyme is a critical regulator of cellular polyamine levels due to its effect on polyamine transport and its ability to target ornithine decarboxylase for degradation. Antizyme expression is autoregulatory, through dependence on an unusual +1 translational frameshift mechanism that responds to polyamine levels. RESULTS: HEK293 cells were depleted of polyamines by treatment with an ornithine decarboxylase inhibitor, difluoromethylornithine (DFMO), and grown in the presence or absence of exogenous polyamines prior to the analysis of ribosomal frameshifting levels. Results obtained using an optimized dual luciferase assay system reveal a 10-fold dynamic range of frameshifting, which correlates positively with polyamine addition. Polyamine addition to cells, which have not been pre-treated with DFMO, also resulted in an increase in antizyme frameshifting but to a lesser degree (1.3 to 1.5-fold). In addition, the constructs with the 3' deletion were more responsive to stimulation by polyamine addition than those with the 5' deletion. CONCLUSIONS: The observed regulation of antizyme frameshifting demonstrates the efficiency of a polyamine homeostatic mechanism, and illustrates the utility of a quantifiable cell-based assay for the analysis of polyamines or their analogues on translational frameshifting. PMID- 11733032 TI - Expression of human and mouse genes encoding polkappa: testis-specific developmental regulation and AhR-dependent inducible transcription. AB - BACKGROUND: Human polkappa is a newly identified low-fidelity DNA polymerase. While the enzyme bypasses an abasic site and acetylaminofluorene-adduct in an error-prone manner, it bypasses benzo[a]pyrene-N2-dG lesions in a mostly error free manner by incorporating predominantly dC opposite the bulky lesions. Benzo[a]pyrene (B[a]P) is activated through intracellular process mediated by the arylhydrocarbon receptor (AhR, also called the dioxin receptor), which is a ligand-activated transcription factor with high affinities for aromatic compounds such as B[a]P and dioxin. RESULTS: We examined promoter structures of the human POLK and mouse Polk genes to study how their expressions are regulated. The mouse Polk gene is developmentally regulated in testis and utilizes two transcription start sites during spermatogenesis, while it utilizes only one site in tissues other than testis. Both of the mouse Polk and the human POLK genes have two AhR binding sites in the promoter regions and the expression of the mouse Polk gene is indeed enhanced upon AhR-activation. CONCLUSIONS: The AhR activation increases expression of the mouse Polk gene and probably the human POLK gene, the product of which bypasses benzo[a]pyrene-N2-dG lesions in a mostly accurate manner. Thus, polkappa seems to function to reduce mutagenesis at benzo[a]pyrene-adducts, although it may also have a role related to spermatogenesis. PMID- 11733033 TI - Herpes simplex virus encodes a virion-associated protein which promotes long cellular processes in over-expressing cells. AB - BACKGROUND: Herpes simplex virus (HSV) possesses a number of accessory genes which are dispensable for replication in cell culture. A previous study showed that the UL21 gene product of HSV type 1 is a virion component that is not necessary for viral replication. The function of the gene product remains unknown. RESULTS: We found that the HSV-1 UL21 gene product, a capsid-associated tegument protein with an apparent molecular mass of 62 kDa, promotes the outgrowth of long cellular processes when it is over-expressed in non-neural cells. The UL21 protein co-localizes and physically associates with microtubules in the long processes. Analysis using mutant proteins implicates a proline-rich region in promotion of the processes. CONCLUSIONS: The results suggest that the UL21 protein, like tau and other MAPs, promotes the process by directly or indirectly interacting with microtubules and facilitates the intracellular transport of the virus. PMID- 11733034 TI - The mouse Snrpn minimal promoter and its human orthologue: activity and imprinting. AB - BACKGROUND: Microdeletions in chromosome 15q13-15 of Prader-Willi (PWS) and Angelman Syndrome (AS) patients suggested that SNRPN promoter/exon 1, together with a short sequence located approximately 35 kb upstream, constitute an imprinting control centre that regulates the entire 2 Mb PWS/AS imprinted domain. We have recently shown that a minitransgene composed of the human upstream sequence and mouse Snrpn promoter/exon 1 harbours all the elements necessary for establishing and maintaining an imprinted state. RESULTS: Here we describe, using transfection experiments, the Snrpn minimal promoter (SMP), being composed of the entire 76 bp exon 1 and 84 bp of upstream sequence. A 7 bp element (SBE) within SMP that, in its unmethylated state binds a specific protein, is absolutely required for promoter activity. The orthologous human sequence, in spite of the fact that it possesses an identical SBE, failed to display promoter activity in transfection experiments and failed to create a methylated state of the maternal allele. Transgenic experiments reveal that a mutation in SBE of the mouse sequence did not completely abolish methylation of the maternal allele, indicating that sequences outside SBE participate in this process. Replacement of human exon 1 with the mouse orthologue replenished promoter activity, but left the maternal allele in the transgenic experiment unmethylated. The reciprocal chimera, in which mouse exon 1 was replaced by the human orthologue resulted in loss of promoter activity and did not support differential methylation. CONCLUSIONS: The observations made by in vitro and in vivo experiments suggest that several cis elements which are involved in Snrpn promoter activity and the imprinting process are present in the mouse promoter and absent in the human orthologous sequence. PMID- 11733036 TI - Mammalian homologue of E. coli Ras-like GTPase (ERA) is a possible apoptosis regulator with RNA binding activity. AB - BACKGROUND: ERA (Escherichia coli Ras-like protein) is an E. coli GTP binding protein that is essential for proliferation. A DNA database search suggests that homologous sequences with ERA exist in various organisms including human, mouse, Drosophila, Caenorhabditis elegans and Antirrhinum majus. However, the physiological function of eukaryotic ERA-like proteins is not known. RESULTS: We have cloned cDNAs encoding the entire coding region of a human homologue (H-ERA) and a mouse homologue (M-ERA) of ERA. The mammalian homologue of ERA consists of a typical GTPase/GTP-binding domain and a putative K homology (KH) domain, which is known as an RNA binding domain. We performed transfection experiments with wild-type H-ERA or various H-ERA mutants. H-ERA possessing the amino acid substitution mutation into the GTPase domain induced apoptosis of HeLa cells, which was blocked by Bcl-2 expression. Deletion of the C-terminus, which contains a part of the KH domain, alleviated apoptosis by the H-ERA mutant, suggesting the importance of this domain in the function of H-ERA. We have also shown the RNA binding activity of H-ERA by pull-down experiments using RNA homopolymer immobilized on beads or recombinant H-ERA proteins. CONCLUSION: Our data suggest that H-ERA plays an important role in the regulation of apoptotic signalling with its GTPase/GTP binding domain. PMID- 11733035 TI - Math6, a bHLH gene expressed in the developing nervous system, regulates neuronal versus glial differentiation. AB - BACKGROUND: Whereas multiple basic helix-loop-helix (bHLH) genes are expressed in the developing nervous system, they account for the differentiation of only subsets of neurones, suggesting that there may be as-yet unidentified bHLH genes. RESULTS: We have isolated a novel bHLH gene, designated Math6, a distant mammalian homologue of the Drosophila proneural gene atonal. Structural analysis of the Math6 gene demonstrated that the coding region is divided into three exons, whereas that of other atonal homologues is present in a single exon, indicating that the genomic structure of Math6 is unique among the atonal homologues. Math6 is initially expressed by neural precursor cells in the ventricular zone, but later by subsets of differentiating and mature neurones such as hippocampal neurones and cerebellar Purkinje cells. Mis-expression of Math6 with retrovirus in the developing retina induced neurogenesis, while inhibiting gliogenesis, without affecting cell proliferation and death. CONCLUSIONS: These results show that cells which would normally differentiate into glia adopted the neuronal fate by mis-expression of Math6, indicating that Math6 promotes neuronal vs. glial fate determination in the nervous system. PMID- 11733037 TI - Distinct regulatory mechanism for p70 S6 kinase beta from that for p70 S6 kinase alpha. AB - BACKGROUND: A novel ribosomal S6 kinase, termed p70 S6 kinase beta (p70beta), has a highly homologous amino acid sequence to that of p70/p85 S6 kinase (p70alpha). This includes the critical phosphorylation sites, Thr252, Ser394 and Thr412 in p70alpha1, which correspond to Thr241, Ser383 and Thr401 in p70beta1, respectively. However, the regulatory mechanism for p70beta remains to be elucidated. RESULTS: We report here the expression and the mechanism of in vivo regulation of p70beta. Two isoforms, p70beta1 and p70beta2, were expressed in a variety of tissues at a different level. p70beta1 was mainly targeted to the nucleus, whereas p70beta2 dispersed throughout the cytoplasm including nucleoplasm. The kinase activity of p70beta1 was less sensitive to the inhibition induced by rapamycin, wortmannin and amino acid withdrawal than that of p70alpha. The portion of p70beta activity inhibited by rapamycin was rescued by the rapamycin-resistant mutant of the mammalian target of rapamycin (mTOR). Mutational analysis revealed that the phosphorylation of Thr241 and Thr401 in p70beta1 was indispensable for the kinase activity. In contrast, a p70beta1 mutant in which Ser383 was substituted with Gly (S383G) still retained nearly the half maximal activity. Sequential phosphorylation of wild-type and S383G mutant of p70beta1 with mTOR and 3-phosphoinositide-dependent protein kinase 1 (PDK1) in vitro synergistically activated their kinase activities. CONCLUSION: These results indicate that p70beta is regulated by the mTOR- and PDK1-signalling pathways through a synergistic interaction between phosphorylated Thr241 and Thr401, while Ser383 plays minor role in their activation mechanism. Activated p70beta may be less sensitive to dephosphorylation mediated by putative phosphatases activated by rapamycin, amino acid withdrawal, and probably wortmannin. PMID- 11733040 TI - Probing the machinery of intracellular trafficking with the atomic force microscope. AB - Atomic force microscopy has emerged as a powerful tool for characterizing single biological macromolecules, macromolecular assemblies, and whole cells in aqueous buffer, in real time, and at molecular-scale spatial and force resolution. Many of the central elements of intracellular transport are tens to hundreds of nanometers in size and highly dynamic. Thus, atomic force microscopy provides a valuable means of addressing questions of structure and mechanism in intracellular transport. We begin this review of recent efforts to apply atomic force microscopy to problems in intracellular transport by discussing the technical principles behind atomic force microscopy. We then turn to three specific areas in which atomic force microscopy has been applied to problems with direct implications for intracellular trafficking: cytoskeletal structure and dynamics, vesicular transport, and receptor-ligand interactions. In each case, we discuss studies which use both intact cellular elements and reconstituted models. While many technical challenges remain, these studies point to several areas where atomic force microscopy can be used to provide valuable insight into intracellular transport at exquisite spatial and energetic resolution. PMID- 11733041 TI - Using optics to measure biological forces and mechanics. AB - Spanning all size levels, regulating biological forces and transport are fundamental life processes. Used by various investigators over the last dozen years, optical techniques offer unique advantages for studying biological forces. The most mature of these techniques, optical tweezers, or the single-beam optical trap, is commercially available and is used by numerous investigators. Although technical innovations have improved the versatility of optical tweezers, simple optical tweezers continue to provide insights into cell biology. Two new, promising optical technologies, laser-tracking microrheology and the optical stretcher, allow mechanical measurements that are not possible with optical tweezers. Here, I review these various optical technologies and their roles in understanding mechanical forces in cell biology. PMID- 11733042 TI - Total internal reflection fluorescence microscopy in cell biology. AB - Key events in cellular trafficking occur at the cell surface, and it is desirable to visualize these events without interference from other regions deeper within. This review describes a microscopy technique based on total internal reflection fluorescence which is well suited for optical sectioning at cell-substrate regions with an unusually thin region of fluorescence excitation. The technique has many other applications as well, most notably for studying biochemical kinetics and single biomolecule dynamics at surfaces. A brief summary of these applications is provided, followed by presentations of the physical basis for the technique and the various ways to implement total internal reflection fluorescence in a standard fluorescence microscope. PMID- 11733043 TI - Applying multiphoton imaging to the study of membrane dynamics in living cells. AB - The endomembrane system of a cell is a highly dynamic, ephemeral structure that is difficult to visualize. Reconstructions from sections of fixed material can provide high-resolution information on intercellular membrane architecture, but such techniques are fraught with artifacts and are of little help in understanding the dynamics of intracellular membrane traffic. Recently, the availability of fluorescent membrane probes and the development of techniques for optically sectioning intact specimens have allowed glimpses of membrane dynamics to be visualized in living tissue. In this review we discuss the potential of a new optical sectioning technique, multiphoton imaging, for visualizing membrane dynamics in living cells. Multiphoton microscopy offers an unparalleled ability to obtain images from deep within specimens while minimizing the effects of phototoxicity. PMID- 11733044 TI - Achieving molecular selectivity in imaging using multiphoton Raman spectroscopy techniques. AB - In the case of most optical imaging methods, contrast is generated either by physical properties of the sample (Differential Image Contrast, Phase Contrast), or by fluorescent labels that are localized to a particular protein or organelle. Standard Raman and infrared methods for obtaining images are based upon the intrinsic vibrational properties of molecules, and thus obviate the need for attached fluorophores. Unfortunately, they have significant limitations for live cell imaging. However, an active Raman method, called Coherent Anti-Stokes Raman Scattering (CARS), is well suited for microscopy, and provides a new means for imaging specific molecules. Vibrational imaging techniques, such as CARS, avoid problems associated with photobleaching and photo-induced toxicity often associated with the use of fluorescent labels with live cells. Because the laser configuration needed to implement CARS technology is similar to that used in other multiphoton microscopy methods, such as two-photon fluorescence and harmonic generation, it is possible to combine imaging modalities, thus generating simultaneous CARS and fluorescence images. A particularly powerful aspect of CARS microscopy is its ability to selectively image deuterated compounds, thus allowing the visualization of molecules, such as lipids, that are chemically indistinguishable from the native species. PMID- 11733045 TI - Fluorescence correlation spectroscopy measures molecular transport in cells. AB - Fluorescence correlation spectroscopy (FCS) can measure dynamics of fluorescent molecules in cells. FCS measures the fluctuations in the number of fluorescent molecules in a small volume illuminated by a thin beam of excitation light. These fluctuations are processed statistically to yield an autocorrelation function from which rates of diffusion, convection, chemical reaction, and other processes can be extracted. The advantages of this approach include the ability to measure the mobility of a very small number of molecules, even down to the single molecule level, over a wide range of rates in very small regions of a cell. In addition to rates of diffusion and convection, FCS also provides unique information about the local concentration, states of aggregation and molecular interaction using fluctuation amplitude and cross-correlation methods. Recent advances in technology have rendered these once difficult measurements accessible to routine use in cell biology and biochemistry. This review provides a summary of the FCS method and describes current areas in which the FCS approach is being extended beyond its original scope. PMID- 11733046 TI - Near-field scanning optical microscopy, a siren call to biology. AB - Near-field illumination of a sample with visible light can resolve features well beyond the resolution of conventional, far-field microscopes. Near-field scanning optical microscopy (NSOM) then has the potential of extending the resolution of techniques such as fluorescent labeling, yielding images of cell structures and molecules on the nanoscale. However, major problems remain to be solved before NSOM can be easily used for wet biological samples. The most significant of these is control of the distance between near-field aperture and the sample surface. Hence, while NSOM promises much, its application to biology is about where electron microscopy was 40 or 50 years ago. PMID- 11733047 TI - Identification of a conserved loop in Mog1 that releases GTP from Ran. AB - Ran regulates nuclear import and export pathways by coordinating the assembly and disassembly of transport complexes. These transport reactions are linked to the GTPase cycle and subcellular distribution of Ran. Mog1 is an evolutionarily conserved nuclear protein that binds RanGTP and stimulates guanine nucleotide release, suggesting Mog1 regulates the nuclear transport functions of Ran. In the present study, we have characterized the nuclear import pathway of Mog1, and we have defined the domain in Mog1 that stimulates GTP release from Ran. In permeabilized cells, nuclear import of Mog1 is independent of exogenously added factors, and is inhibited by wheat germ agglutinin, indicating that translocation of Mog1 involves physical interactions with the nuclear pore complex. In contrast to RanGEF, which is restricted to the nucleus, Mog1 shuttles between the nucleus and the cytoplasm. Single-point mutations in acidic residues of Mog1 (Asp25, Asp34, Glu37) dramatically reduce GTP release and Ran binding activity, whereas mutation of a single basic residue (Arg30) renders Mog1 hyperactive for GTP release. These mutations map within a conserved, solvent-exposed loop in Mog1 that is functionally similar to the beta-wedge used by RanGEF to promote nucleotide release from Ran. These data suggest that Mog1 and RanGEF use similar mechanisms to facilitate guanine nucleotide release from Ran. PMID- 11733048 TI - Dynamics of leptomycin B-sensitive nucleocytoplasmic flux of parathyroid hormone related protein. AB - Parathyroid hormone-related protein is responsible for hypercalcemia induced by various tumors. The similarity of its N-terminus to that of parathyroid hormone enables parathyroid hormone-related protein to share parathyroid hormone's signaling properties, but the rest of the molecule possesses distinct functions including a role in the nucleus/nucleolus in reducing apoptosis and enhancing cell proliferation. We have previously shown that parathyroid hormone-related protein nuclear import is mediated by importin beta1. Here we use fluorescence recovery after photobleaching for the first time to show that, in living cells, parathyroid hormone-related protein is exported from the nucleus in a leptomycin B-sensitive manner, implicating CRM1 as the parathyroid hormone-related protein nuclear export receptor. Leptomycin B treatment significantly reduced the rate of nuclear export 4 -10-fold, thereby increasing parathyroid hormone-related protein concentration in the nucleus/nucleolus about 2-fold. Intriguingly, this also led to a 2-fold reduced nuclear import rate. Inhibiting the nuclear export of a protein able to shuttle between nucleus and cytoplasm through distinct receptors thus can also affect nuclear import, indicating that the subcellular localization of a protein containing distinct nuclear import and export signals is the product of an integrated system. Although there have been several recent studies examining the dynamics of intranuclear transport using fluorescence recovery after photobleaching, this represents, to our knowledge, the first use of the technique to examine the kinetics of nucleocytoplasmic flux in living cells. PMID- 11733049 TI - Sgf1p, a new component of the Sec34p/Sec35p complex. AB - Here we report the identification of SGF1 as a high-copy suppressor of the sec35 1 mutant. SGF1 encodes an essential hydrophilic protein of approximately 100 kDa. Using the yeast two-hybrid system and coprecipitation studies, we demonstrate that Sgf1p is a new subunit of the multiprotein Sec34p/Sec35p complex. Reduced levels of Sgf1p lead to the accumulation of a variety of membranes as well as a kinetic block in endoplasmic reticulum to Golgi traffic. Immunofluorescence studies demonstrate that Sec34p is found throughout the Golgi, with a high concentration on early Golgi. Although an earlier study suggested that Sec34p (Grd20p) is not required for protein secretion, we show here that the sec34-2 and sec35-1 mutations lead to a pleiotropic block in the secretion of all proteins into the growth medium. PMID- 11733050 TI - The ER to Golgi interface is the major concentration site of secretory proteins in the exocrine pancreatic cell. AB - By using quantitative immuno-electron microscopy of two-sided labeled resin sections of rat exocrine pancreatic cells, we have established the relative concentrations of the secretory proteins amylase and chymotrypsinogen in the compartments of the secretory pathway. Their total concentration over the entire pathway was approximately 11 and approximately 460 times, respectively. Both proteins exhibited their largest increase in concentration between the endoplasmic reticulum and cis-Golgi, where they were concentrated 3-4 and 50-70 times, respectively. Over the further pathway, increases in concentration were moderate, albeit two times higher for chymotrypsinogen than for amylase. From trans-Golgi to secretory granules, where the main secretory protein concentration is often thought to occur, relatively small concentration increases were observed. Additional observations on a third secretory protein, procarboxypeptidase A, showed a concentration profile very similar to chymotrypsinogen. The relatively high concentration of amylase in the early compartments of the secretory route is consistent with its exceptionally slow intracellular transport. Our data demonstrate that secretory proteins undergo their main concentration between the endoplasmic reticulum and cis-Golgi, where we have previously found concentration activity associated with vesicular tubular clusters (Martinez-Menarguez JA, Geuze HJ, Slot JW, Klumperman J. Cell 1999; 98: 81-90). PMID- 11733051 TI - Drosophila Amphiphysin is a post-synaptic protein required for normal locomotion but not endocytosis. AB - Clathrin-mediated endocytosis is required to recycle synaptic vesicles for fast and efficient neurotransmission. Amphiphysins are thought to be multiprotein adaptors that may contribute to this process by bringing together many of the proteins required for endocytosis. Their in vivo function, however, has yet to be determined. Here, we show that the Drosophila genome encodes a single amphiphysin gene that is broadly expressed during development. We also show that, unlike its vertebrate counterparts, Drosophila Amphiphysin is enriched postsynaptically at the larval neuromuscular junction. To determine the role of Drosophila Amphiphysin, we also generated null mutants which are viable but give rise to larvae and adults with pronounced locomotory defects. Surprisingly, the locomotory defects cannot be accounted for by alterations in the morphology or physiology of the neuromuscular junction. Moreover, using stimulus protocols designed to test endocytosis under moderate and extreme vesicle cycling, we could not detect any defect in the neuromuscular junction of the amphiphysin mutant. Taken together, our findings suggest that Amphiphysin is not required for viability, nor is it absolutely required for clathrin-mediated endocytosis. However, Drosophila Amphiphysin function is required in both larvae and adults for normal locomotion. PMID- 11733052 TI - Clathrin hub expression dissociates the actin-binding protein Hip1R from coated pits and disrupts their alignment with the actin cytoskeleton. AB - The actin cytoskeleton has been implicated in the maintenance of discrete sites for clathrin-coated pit formation during receptor-mediated endocytosis in mammalian cells, and its function is intimately linked to the endocytic pathway in yeast. Here we demonstrate that staining for mammalian endocytic clathrin coated pits using a monoclonal antibody against the AP2 adaptor complex revealed a linear pattern that correlates with the organization of the actin cytoskeleton. This vesicle organization was disrupted by treatment of cells with cytochalasin D, which disassembles actin, or with 2,3-butanedione monoxime, which prevents myosin association with actin. The linear AP2 staining pattern was also disrupted in HeLa cells that were induced to express the Hub fragment of the clathrin heavy chain, which acts as a dominant-negative inhibitor of receptor-mediated endocytosis by direct interference with clathrin function. Additionally, Hub expression caused the actin-binding protein Hip1R to dissociate from coated pits. These findings indicate that proper function of clathrin is required for coated pit alignment with the actin cytoskeleton and suggest that the clathrin-Hip1R interaction is involved in the cytoskeletal organization of coated pits. PMID- 11733053 TI - The fuss about Mus81. AB - Endonucleolytic cleavage of Holliday junctions is important in recombination and replication. Mus81 proteins in yeasts and humans appear to have many, but not all, of the expected properties of eukaryotic Holliday junction resolvases, with intriguing connections to DNA replication checkpoints. PMID- 11733054 TI - RNA targets of the fragile X protein. AB - Three papers published recently in Cell bring the power of human genetics, Drosophila genetics, and genomics to bear on the understanding of fragile X syndrome. They provide further support for the importance of local protein synthesis within a neuron as a determinant of proper synaptogenesis and the development of cognitive abilities. PMID- 11733055 TI - Recognizing phosphatidylinositol 3-phosphate. AB - Phosphatidylinositol 3-phosphate directs the endosomal localization of regulatory proteins by binding to FYVE and PX domains. New structures of these domains complexed with the phosphoinositide headgroup show how interactions with phosphate and hydroxyl groups differentiate this lipid from all others. PMID- 11733056 TI - New glimpses of an old machine. AB - Two recent studies illustrate the ability of the Mre11/Rad50 DNA repair complex to bind and connect DNA ends. Specific stimulation of DNA ligase IV-mediated end joining by Mre11 complexes from S. cerevisiae suggests the possibility of a direct role in nonhomologous end-joining in eukaryotic cells. PMID- 11733057 TI - Microarray analysis and organization of circadian gene expression in Drosophila. AB - We have used high-density oligonucleotide arrays to study global circadian gene expression in Drosophila melanogaster. Coupled with an analysis of clock mutant (Clk) flies, a cell line designed to identify direct targets of the CLOCK (CLK) transcription factor and differential display, we uncovered several striking features of circadian gene networks. These include the identification of 134 cycling genes, which contribute to a wide range of diverse processes. Many of these clock or clock-regulated genes are located in gene clusters, which appear subject to transcriptional coregulation. All oscillating gene expression is under clk control, indicating that Drosophila has no clk-independent circadian systems. An even larger number of genes is affected in Clk flies, suggesting that clk affects other genetic networks. As we identified a small number of direct target genes, the data suggest that most of the circadian gene network is indirectly regulated by clk. PMID- 11733058 TI - Comprehensive analysis of photoreceptor gene expression and the identification of candidate retinal disease genes. AB - To identify the full set of genes expressed by mammalian rods, we conducted serial analysis of gene expression (SAGE) by using libraries generated from mature and developing mouse retina. We identified 264 uncharacterized genes that were specific to or highly enriched in rods. Nearly half of all cloned human retinal disease genes are selectively expressed in rod photoreceptors. In silico mapping of the human orthologs of genes identified in our screen revealed that 86 map within intervals containing uncloned retinal disease genes, representing 37 different loci. We expect these data will allow identification of many disease genes, and that this approach may be useful for cloning genes involved in classes of disease where cell type-specific expression of disease genes is observed. PMID- 11733059 TI - Drosophila fragile X-related gene regulates the MAP1B homolog Futsch to control synaptic structure and function. AB - Fragile X mental retardation gene (FMR1) encodes an RNA binding protein that acts as a negative translational regulator. We have developed a Drosophila fragile X syndrome model using loss-of-function mutants and overexpression of the FMR1 homolog (dfxr). dfxr nulls display enlarged synaptic terminals, whereas neuronal overexpression results in fewer and larger synaptic boutons. Synaptic structural defects are accompanied by altered neurotransmission, with synapse type-specific regulation in central and peripheral synapses. These phenotypes mimic those observed in mutants of microtubule-associated Futsch. Immunoprecipitation of dFXR shows association with futsch mRNA, and Western analyses demonstrate that dFXR inversely regulates Futsch expression. dfxr futsch double mutants restore normal synaptic structure and function. We propose that dFXR acts as a translational repressor of Futsch to regulate microtubule-dependent synaptic growth and function. PMID- 11733060 TI - Remodeling of synaptic actin induced by photoconductive stimulation. AB - Use-dependent synapse remodeling is thought to provide a cellular mechanism for encoding durable memories, yet whether activity triggers an actual structural change has remained controversial. We use photoconductive stimulation to demonstrate activity-dependent morphological synaptic plasticity by video imaging of GFP-actin at individual synapses. A single tetanus transiently moves presynaptic actin toward and postsynaptic actin away from the synaptic junction. Repetitive spaced tetani induce glutamate receptor-dependent stable restructuring of synapses. Presynaptic actin redistributes and forms new puncta that label for an active synapse marker FM5-95 within 2 hr. Postsynaptic actin sprouts projections toward the new presynaptic actin puncta, resembling the axon-dendrite interaction during synaptogenesis. Our results indicate that activity-dependent presynaptic structural plasticity facilitates the formation of new active presynaptic terminals. PMID- 11733061 TI - Forebrain-specific calcineurin knockout selectively impairs bidirectional synaptic plasticity and working/episodic-like memory. AB - Calcineurin is a calcium-dependent protein phosphatase that has been implicated in various aspects of synaptic plasticity. By using conditional gene-targeting techniques, we created mice in which calcineurin activity is disrupted specifically in the adult forebrain. At hippocampal Schaffer collateral-CA1 synapses, LTD was significantly diminished, and there was a significant shift in the LTD/LTP modification threshold in mutant mice. Strikingly, although performance was normal in hippocampus-dependent reference memory tasks, including contextual fear conditioning and the Morris water maze, the mutant mice were impaired in hippocampus-dependent working and episodic-like memory tasks, including the delayed matching-to-place task and the radial maze task. Our results define a critical role for calcineurin in bidirectional synaptic plasticity and suggest a novel mechanistic distinction between working/episodic like memory and reference memory. PMID- 11733062 TI - The overall pattern of cardiac contraction depends on a spatial gradient of myosin regulatory light chain phosphorylation. AB - Evolution of the human heart has incorporated a variety of successful strategies for motion used throughout the animal kingdom. One such strategy is to add the efficiency of torsion to compression so that blood is wrung, as well as pumped, out of the heart. Models of cardiac torsion have assumed uniform contractile properties of muscle fibers throughout the heart. Here, we show how a spatial gradient of myosin light chain phosphorylation across the heart facilitates torsion by inversely altering tension production and the stretch activation response. To demonstrate the importance of cardiac light chain phosphorylation, we cloned a myosin light chain kinase from a human heart and have identified a gain-in-function mutation in two individuals with cardiac hypertrophy. PMID- 11733063 TI - A signaling adapter function for alpha6beta4 integrin in the control of HGF dependent invasive growth. AB - alpha6beta4 integrin and the Met receptor for HGF have been shown independently to promote invasive growth. We demonstrate here that Met selectively associates with alpha6beta4. In carcinoma cells expressing Met alone, HGF does not exert significant biological effects. Ectopic expression of alpha6beta4 restores HGF regulated processes. Following Met activation, alpha6beta4 is tyrosine phosphorylated and combines with Shc and PI3K, generating an additional signaling platform that potentiates HGF-triggered activation of Ras- and PI3K-dependent pathways. In the presence of an alpha6beta4 mutant defective for Shc recruitment, Met cannot sustain HGF-mediated responses. Surprisingly, a truncated beta4 unable to bind laminins retains the activity of wild-type alpha6beta4. Such findings invoke an unexpected role for alpha6beta4 in cancer invasion as a functional amplifier of biochemical outputs rather than a mechanical adhesive device. PMID- 11733064 TI - Regulation of the Bub2/Bfa1 GAP complex by Cdc5 and cell cycle checkpoints. AB - During mitosis, a ras-related GTPase (Tem1) binds GTP and activates a signal transduction pathway to allow mitotic exit. During most of the cell cycle, Tem1 function is antagonized by a GTPase-activating protein complex, Bfa1/Bub2. How the Bfa1/Bub2 complex is regulated is not well understood. We find that Polo/Cdc5 kinase acts upstream of Bfa1/Bub2 in the mitotic exit network. Cdc5 phosphorylates Bfa1 and acts to antagonize Bfa1 function to promote mitotic exit. Bfa1 is regulated by multiple cell cycle checkpoints. The spindle assembly and spindle orientation checkpoints inhibit Bfa1 phosphorylation. DNA damage does not inhibit Bfa1 phosphorylation and instead causes a Rad53- and Dun1-dependent modification of Bfa1. Regulation of Bfa1 may therefore be a key step controlled by multiple checkpoint pathways to ensure a mitotic arrest. PMID- 11733065 TI - Mobilization of processed, membrane-tethered SPT23 transcription factor by CDC48(UFD1/NPL4), a ubiquitin-selective chaperone. AB - The OLE pathway of yeast regulates the level of the ER-bound enzyme Delta9-fatty acid desaturase OLE1, thereby controlling membrane fluidity. A central component of this regulon is the transcription factor SPT23, a homolog of mammalian NF kappaB. SPT23 is synthesized as an inactive, ER membrane-anchored precursor that is activated by regulated ubiquitin/proteasome-dependent processing (RUP). We now show that SPT23 dimerizes prior to processing and that the processed molecule, p90, retains its ubiquitin modification and initially remains tethered to its unprocessed, membrane-bound SPT23 partner. Subsequently, p90 is liberated from its partner for nuclear targeting by the activity of the chaperone-like CDC48(UFD1/NPL4) complex. Remarkably, this enzyme binds preferentially ubiquitinated substrates, suggesting that CDC48(UFD1/NPL4) is qualified to selectively remove ubiquitin conjugates from protein complexes. PMID- 11733066 TI - High resolution structure of the large ribosomal subunit from a mesophilic eubacterium. AB - We describe the high resolution structure of the large ribosomal subunit from Deinococcus radiodurans (D50S), a gram-positive mesophile suitable for binding of antibiotics and functionally relevant ligands. The over-all structure of D50S is similar to that from the archae bacterium Haloarcula marismortui (H50S); however, a detailed comparison revealed significant differences, for example, in the orientation of nucleotides in peptidyl transferase center and in the structures of many ribosomal proteins. Analysis of ribosomal features involved in dynamic aspects of protein biosynthesis that are partially or fully disordered in H50S revealed the conformations of intersubunit bridges in unbound subunits, suggesting how they may change upon subunit association and how movements of the L1-stalk may facilitate the exit of tRNA. PMID- 11733068 TI - Detection of DNA adducts using a quantitative long PCR technique and the fluorogenic 5' nuclease assay (TaqMan). AB - The detection of DNA adducts is an important component in assessing the mutagenic potential of exogenous and endogenous compounds. Here, we report an in vitro quantitative long PCR (XL-PCR) assay to measure DNA adducts in human genomic DNA based on their ability to block and inhibit PCR amplification. Human genomic DNA was exposed to test compounds and then a target sequence was amplified by XL-PCR. The amplified sequence was then quantified using fluorogenic 5' nuclease PCR (TaqMan) and normalized to a solvent-treated control. The extent of DNA adduction was determined based on the reduction in amplification of the target sequence in the treated sample. A 17.7kb beta-globin fragment was chosen as the target sequence for these studies, since preliminary experiments revealed a two-fold increased sensitivity of this target compared to a 10.4kb HPRT fragment for detecting hydrogen peroxide-induced DNA damage. Validation of the XL-PCR assay with various compounds demonstrated the versatility of the assay for detecting a wide range of adducts formed by direct acting or S9-activated mutagens. The same DNA samples were also analyzed using 32P-postlabeling techniques (thin-layer chromatography or high-performance liquid chromatography) to confirm the presence of DNA adducts and estimate their levels. Whereas 32P-postlabeling with nuclease P(1) enrichment was more sensitive for detecting bulky adducts induced by the compounds benzo[a]pyrene, dimethylbenzanthracene, 3-methylindole, indole 3 carbinol, or 2-acetylaminofluorene, the XL-PCR procedure was more sensitive for detecting smaller or labile DNA adducts formed by the compounds methyl methanesulfonate, diethyl nitrosamine, ethylnitrosourea, diepoxybutane, ICR-191, styrene oxide, or aflatoxin B(1). Compounds not expected to form adducts in DNA, such as clofibrate, phenobarbital, chloroform or acetone, did not produce a positive response in the XL-PCR assay. Thus, quantitative XL-PCR provides a rapid, high-throughput assay for detecting DNA damage that complements the existing 32P-postlabeling assay with nuclease P(1) enrichment. PMID- 11733069 TI - Cellular determinants of the mutational specificity of 1-nitroso-6-nitropyrene and 1-nitroso-8-nitropyrene in the lacI gene of Escherichia coli. AB - We have characterized 202 lacI(-) mutations, and 158 dominant lacI(-d) mutations following treatment of Escherichia coli strains NR6112 and EE125 with 1-nitroso-6 nitropyrene (1,6-NONP), an activated metabolite of the carcinogen 1,6 dinitropyrene. In all, 91% of the induced point mutations occurred at G:C residues. The -(G:C) frameshifts were the dominant mutational class in the lacI( ) collections of both NR6112 and EE125, and in the lacI(-d) collection of NR6112. Frameshift mutations occurred preferentially in runs of guanine residues, and their frequency increased with the length of the reiterated sequence. In strain EE125, which contained the plasmid pKM101, there was a marked stimulation in the frequency of base substitution mutations that was particularly apparent in the lacI(-d) collection. This study completes a comprehensive analysis of 1194 lacI( ) and 348 lacI(-d) mutations induced by either 1,6-NONP or its positional isomer 1-nitroso-8-nitropyrene (1,8-NONP) in strains of E. coli that differ with regard to their ability to carry out nucleotide excision repair and/or their ability to express the translesion synthesis DNA polymerase RI (MucAB) encoded by plasmid pKM101. Among the mutations are 763 frameshift mutations, 367 base substitutions and 47 deletions; these mutations have been characterized at more than 300 distinct sites in the lacI gene. Our studies provide detailed insight into the DNA sequence alterations and mutational mechanisms associated with dinitropyrene mutagenesis. We review the mutational spectra, and discuss cellular lesion repair or tolerance mechanisms that modulate the observed mutational specificity. PMID- 11733070 TI - Prevalence of mutagens in the environment: experimental data versus simulations. AB - The recent availability of experimental data on the mutagenicity in Salmonella of a subset of high production volume chemicals allowed a comparison with the estimate derived from computer-based simulations. The prevalence of mutagens in the subsets was not significantly different: 20% versus 19.5% based upon experimental and simulation data, respectively. This provides support for the use of the rapid and low-cost simulation approach. PMID- 11733071 TI - Alteration of a sequence with homology to human endogenous retrovirus (HERV-K) in primary human glioma: implications for viral repeat mediated rearrangement. AB - We had earlier demonstrated that a comparison of DNA fingerprinting profiles of tumor and corresponding normal DNA from the same patient by random amplified polymorphic DNA (RAPD) analysis can readily demonstrate alterations in tumor DNA [Gene 206 (1998) 45 and J. Neuro Oncol. 48 (2000) 1]. These alterations could be used to identify changes in tumor DNA where the prior identity of the locus was not known. In this study, we report the identification, cloning and characterization of a RAPD amplified fragment which was lost in a glioma, a grade IV glioblastoma multiforme (GBM). Comparison of the RAPD profile of tumor and corresponding leucocyte DNA revealed several differences between the two. These included a band of 443 bases, which was demonstrated in the normal, but not in tumor DNA. On sequencing, this band was found to be homologous with a group of SINE sequences, which are probably derived from the human endogenous retrovirus-K (HERV-K). Homology search also reveals that HERV-K-derived sequences are interspersed, amongst others, in the tumor suppressor gene BRCA2 and the DNA repair gene XRCC1. Of particular interest is the inverted repeat pattern of HERV derived sequences in the genes. While not demonstrating a cause effect relationship, this highlights the possible role of such virus-derived sequences in gene inactivation by recombination during tumorigenesis. PMID- 11733072 TI - Cytochrome b(5) coexpression increases the CYP2E1-dependent mutagenicity of dialkylnitrosamines in methyltransferase-deficient strains of Salmonella typhimurium. AB - Addition of cytochrome b(5) to recombinant cytochrome P450 2E1 systems has been shown to enhance the metabolism of dialkylnitrosamines in vitro. To determine if this effect could be observed with recombinant expression systems in vivo, we have constructed mutagenicity tester strains that coexpress full-length human cytochrome P450 2E1 (CYP2E1), rat cytochrome P450 reductase, and human cytochrome b(5) in Salmonella typhimurium lacking ogt and ada methyltransferases (YG7104, ogt(-); and YG7108, ogt(-), ada(-)). These new recombinant strains exhibit a four to five-fold greater mutagenic response to dimethylnitrosamine, diethylnitrosamine, and dipropylnitrosamine than strains that contain only CYP2E1 and reductase, and are over 100-fold more sensitive to nitrosamines than the parental strains in the presence of an exogenous activating system (S9 fraction). The four-fold increase in mutagenicity in the presence of cytochrome b(5) was consistent with increasing alkyl chain length up to dibutylnitrosamine, which was poorly activated by CYP2E1. The greatest enhancement was obtained with a tricistronic construct in which the b(5) cDNA preceded the P450 and reductase cDNAs; placing the b(5) cDNA after the reductase cDNA was substantially less effective. These new, highly sensitive strains may prove useful in the detection of nitrosamine contamination of food and environmental samples. PMID- 11733073 TI - Analysis of lacI mutations in Big Blue transgenic mice subjected to parasite induced inflammation. AB - Parasite infections have long been associated with specific types of human cancers. Schistosoma hematobium is an inducer of urinary bladder cancer, Helicobacter pylori is a gastric carcinogen, and hepatitis B virus and Opisthorchis viverrini are causative agents of hepatocellular carcinoma. Another liver fluke, Fasciola hepatica, has also been identified as a neoplastic risk agent, primarily in animals. We used F. hepatica-induced inflammation in mice to determine if the presence of an aggressive liver fluke could induce mutagenic events in mammalian tissue. This provides a perspective on the relationship between chronic inflammation and cancer and may be a model for future studies on this complex association. In previous studies using the Big Blue transgenic mouse assay, we demonstrated an increase in lacI mutations in liver cells harvested from mice harboring F. hepatica flukes when compared to uninfected control animals. In these studies, we report on the types of mutations associated with this parasite infection. The observed mutational spectrum roughly corresponded to the spectrum of spontaneous mutations in liver cells when compared to control (uninfected) animals. However, the spectrum of mutations from parasitized animals showed a significant increase in complex changes and multiple mutations (18.2%) when compared to what would be expected from control animals (2.8%). PMID- 11733074 TI - Induction of lacI mutations in Big Blue Rat-2 cells treated with 1-(2 hydroxyethyl)-1-nitrosourea: a model system for the analysis of mutagenic potential of the hydroxyethyl adducts produced by 1,3-bis (2-chloroethyl)-1 nitrosourea. AB - We have investigated the genotoxic effects of 1-(2-hydroxyethyl)-1-nitrosourea (HENU). We have chosen this agent because of its demonstrated ability to produce N7-(2-hydroxyethyl) guanine (N7-HOEtG) and O(6)-(2-hydroxyethyl) 2' deoxyguanosine (O(6)-HOEtdG); two of the DNA alkylation products produced by 1,3 bis (2-chloroethyl)-1-nitrosourea (BCNU). For these studies, we have used the Big Blue Rat-2 cell line that contains a lambda/lacI shuttle vector. Treatment of these cells with HENU produced a dose dependent increase in the levels of N7 HOEtG and O(6)-HOEtdG as quantified by HPLC with electrochemical detection. Treatment of Big Blue Rat-2 cells with either 0, 1 or 5mM HENU resulted in mutation frequencies of 7.2+/-2.2x10(-5), 45.2+/-2.9x10(-5) and 120.3+/-24.4x10( 5), respectively. Comparison of the mutation frequencies demonstrates that 1 and 5mM HENU treatments have increased the mutation frequency by 6- and 16-fold, respectively. This increase in mutation frequency was statistically significant (P<0.001). Sequence analysis of HENU-induced mutations have revealed primarily G:C-->A:T transitions (52%) and a significant number of A:T-->T:A transversions (16%). We propose that the observed G:C-->A:T transitions are produced by the DNA alkylation product O(6)-HOEtdG. These results suggest that the formation of O(6) HOEtdG by BCNU treatment contributes to its observed mutagenic properties. PMID- 11733075 TI - Activation of programmed cell death (apoptosis) by adriamycin in human neoplastic cells. AB - We have studied the occurrence of the apoptosis phenomenon in the action of adriamycin (ADR) on human melanoma cells sensitive (ME18) and resistant (ME18/R) to ADR. The study has shown that the intensity of apoptotic morphological changes noted in melanoma cells depended on the duration of the ADR treatment. We have not observed any positive correlation between the induction of apoptosis and sensitivity to ADR. We have used a fluorescence microscope and flow cytometer to evaluate apoptotic events in cells treated with ADR. PMID- 11733076 TI - Study of adaptive mutations in Salmonella typhimurium by using a super-repressing mutant of a trans regulatory gene purR. AB - Salmonella typhimurium purR encodes a transcriptional repressor regulating gene expression of de novo purine nucleotide biosynthesis. It represses purD gene transcription by binding to the 16-base pair purD operator (PUR box). A S. typhimurium strain carrying a super-repressing mutant of purR, purR(s), has been used as an experimental system to study adaptive mutation. Escherichia coli lac genes were genetically engineered into S. typhimurium chromosome and repressed by purR(s) so that they could be used as an indicator of adaptive mutations in purR(s) or in the purD operator. Mutations in purR(s) or in the purD operator accumulated when the mutant strain was placed on a minimal lactose plate supplemented with 10 microg/ml of adenine during prolonged incubation. These specific mutations reverted the mutant strain from lac(-) to lac(+) phenotype. The lac(+) strains were categorized into the early- and late-arising mutants according to the time for colony appearance. Our genetic studies indicate that (i) Poisson distributed mutations accumulated in the chromosomal regulatory gene purR or the purD operator in very slowly dividing cells under selection; (ii) after about 8 days of selection, the frequency of mutations in purD operator reached the high value of about two mutations per 10(8) cells; (iii) the mutational spectrum in the purD operator during growth was not significantly different from that during selection; (iv) defects in mutL or mutS appeared to have a stronger effect on growth-dependent mutations than on adaptive mutations. PMID- 11733077 TI - Loss of heterozygosity (LOH) in 15q26.2-->ter in breast cancer. PMID- 11733079 TI - PNA for rapid microbiology. AB - The acceptance of rRNA sequence diversity as a criterion for phylogenetic discrimination heralds the transition from microbiological identification methods based on phenotypic markers to assays employing molecular techniques. Robust amplification assays and sensitive direct detection methods are rapidly becoming the standard protocols of microbiology laboratories. The emergence of peptide nucleic acid (PNA) from its status as an academic curiosity to that of a promising and powerful molecular tool, coincides with, and complements, the transition to rapid molecular tests. The unique properties of PNA enable the development of assay formats, which go above and beyond the possibilities of DNA probes. PNA probes targeting specific rRNA sequences of yeast and bacteria with clinical, environmental, and industrial value have recently been developed and applied to a variety of rapid assay formats. Some simply incorporate the sensitivity and specificity of PNA probes into traditional methods, such as membrane filtration and microscopic analysis; others involve recent techniques such as real-time and end-point analysis of amplification reactions. PMID- 11733080 TI - Attachment of Staphylococcus aureus to eukaryotic cells and experimental pitfalls in staphylococcal adherence assays: a critical appraisal. AB - Staphylococcus aureus is a bacterial species with pathogenic potential to both humans and animals. The primary natural niche is said to be the human vestibulum nasi from where bacterial cells may spread to the environment or additional anatomical sites such as the perineum or the hands, where residence is usually transient. Apparently, S. aureus is capable of a precise and balanced interaction with specific types of eukaryotic nasal cells. Although a wide variety of important bacterial ligands and possible eukaryote receptors have been described, the precise mechanisms leading to persistent bacterial colonization and, even more importantly, associated infection have not yet been elucidated in detail. This may be a consequence of the fact that most of the adherence factors have been studied individually in simplified in vitro systems, not taking the complexity of multi-factorial in vivo cell-cell interactions into account. An overall scheme of the initial and sequential interactions leading to S. aureus colonization of eukaryotic cell surfaces has not yet emerged. This review concisely describes the current state of affairs in the multi-disciplinary field of staphylococcal adherence research. Specific emphasis is placed upon the pros and cons of the various artificial, mostly in vitro models employed to study the interaction between bacterial and human or animal cells. PMID- 11733081 TI - Green fluorescent protein-based biosensor for detecting SOS-inducing activity of genotoxic compounds. AB - Increasing levels of environmental pollution demand specific and sensitive methods for detection of genotoxic agents in water, food products and environmental samples. Tests for genotoxicity assessment are often based on biosensor strains that respond to DNA damage induced by chemicals. In the present study, fluorescent reporter Escherichia coli strains have been developed, which contain a plasmid-borne transcriptional fusion between the DNA-damage inducible recA promoter and the green fluorescent protein gene (gfp) or a gene encoding a red-shifted, higher intensity GFP variant (mutant 3). GFP-based biosensors allowed the detection of a dose-dependent response to genotoxic agents such as mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and nalidixic acid (NA). A reporter strain carrying recA'-gfp mutant 3 fusion gave more dramatic and sensitive response than a strain containing the wild-type gfp. These results indicate that recA'-gfp mutant 3-based biosensor is potentially useful for detection of genotoxins. PMID- 11733082 TI - Automated image analysis and in situ hybridization as tools to study bacterial populations in food resources, gut and cast of Lumbricus terrestris L. AB - An image analysis procedure was developed for bacterial cells after staining with the DNA-intercalating dye 4'-6-diamidino-2-phenylindole (DAPI), and after in situ hybridization with Cy3-labeled, rRNA-targeted oligonucleotide probes. DAPI- and Cy3-images were captured separately from the same scenery with a cooled digital video camera with three CCD chips for the basic colors red (R), green (G) and blue (B). Using the appropriate filter sets, images of DAPI-stained cells were captured with the red channel shut down, while Cy3-stained cells were captured with the green and blue channels shut down. DAPI images and Cy3 images were subsequently merged to produce virtual color (RGB)-images. Processing of all color channels allowed to specifically enumerate DAPI-stained and hybridized bacteria, to measure their cell sizes, to subsequently calculate their biovolumes and to estimate their biomass. Using this procedure, significant differences were detected in bacterial populations in food resources, digestive tract and cast of the earthworm L. terrestris L. In leaves, bacteria were on average ten times more abundant and two times larger than in soil. In the digestive tract of L. terrestris, however, numbers and volumes of bacteria were comparable to those in soil indicating the disruption of cells originating from leaves before arriving in the foregut. Passage through the digestive tract of L. terrestris significantly reduced bacterial populations belonging to the alpha-, beta- and gamma-subdivisions of Proteobacteria. While these populations did not recover during incubation of cast, populations of the delta-subdivision of Proteobacteria and the Cytophaga-Flavobacterium cluster of the CFB phylum increased in cast. These results suggest a large impact of passage through the digestive tract of L. terrestris on bacterial community structure and demonstrate the usefulness of our image analysis procedure for the determination of cell sizes and biovolumes and thus biomass of specific bacterial populations in different terrestrial habitats. PMID- 11733083 TI - A polyphasic approach for studying the interaction between Ralstonia solanacearum and potential control agents in the tomato phytosphere. AB - Ralstonia solanacearum biovar 2, the causative agent of brown rot in potato, has been responsible for large crop losses in Northwest Europe during the last decade. Knowledge on the ecological behaviour of R. solanacearum and its antagonists is required to develop sound procedures for its control and eradication in infested fields.A polyphasic approach was used to study the invasion of plants by a selected R. solanacearum biovar 2 strain, denoted 1609, either or not in combination with the antagonistic strains Pseudomonas corrugata IDV1 and P. fluorescens UA5-40. Thus, this study combined plating (spread and drop plate methods), reporter gene technology (gfp mutants) and serological (imunofluorescence colony staining [IFC]) and molecular techniques (fluorescent in situ hybridization [FISH], PCR with R. solanacearum specific primers and PCR DGGE on plant DNA extracts). The behaviour of R. solanacearum 1609 and the two control strains was studied in bulk and (tomato) rhizosphere soil and the rhizoplane and stems of tomato plants. The results showed that an interaction between the pathogen and the control strains at the root surface was likely. In particular, R. solanacearum 1609 CFU numbers were significantly reduced on tomato roots treated with P. corrugata IDV1(chr:gfp1) cells as compared to those on untreated roots. Concomitant with the presence of P. corrugata IDV1(chr:gfp1), plant invasion by the pathogen was hampered, but not abolished.PCR-DGGE analyses of the tomato rhizoplane supported the evidence for antagonistic activity against the pathogen; as only weak R. solanacearum 1609 specific bands were detected in profiles derived from mixed systems versus strong bands in profiles from systems containing only the pathogen. Using FISH, a difference in root colonization was demonstrated between the pathogen and one of the two antagonists, i.e. P. corrugata IDV1(chr:gfp1); R. solanacearum strain 1609 was clearly detected in the vascular cylinder of tomato plants, whereas strain IDV1 was absent.R. solanacearum 1609 cells were also detected in stems of plants that had developed in soils treated with this strain, even in cases in which disease symptoms were absent, indicating the occurrence of symptomless infection. In contrast, strain 1609 cells were not found in stems of several plants treated with either one of the two antagonists. The polyphasic analysis is valuable for testing antagonistic strains for approval as biocontrol agents in agricultural practice. PMID- 11733084 TI - Development of the sexual reproductive cycle of Xanthophyllomyces dendrorhous. AB - Conditions inducing the development of holobasidia with terminal basidiospores in wild-type and astaxanthin mutant strains of Xanthophyllomyces dendrorhous were reexamined. Important factors for the development of holobasidia were the incubation temperature and the medium composition. A temperature of 9 degrees C was demonstrated to enhance holobasidia formation. Minimal growth medium with glucose as sole carbon source at concentrations between 80 and 120 mM, and ammonium nitrate with concentrations of 28 mM gave optimal results. A period of 20 or more days was needed for the formation of holobasidia with basidiospores. Additionally, mutant strains of X. dendrorhous were observed to have different abilities to produce holobasidia and strains obtained after protoplast fusion, which have been called fusant in this study, to have an increased capacity to form holobasidia. PMID- 11733085 TI - Local delivery of mithramycin restores vascular reactivity and inhibits neointimal formation in injured arteries and vascular grafts. AB - Arterial restenosis is responsible for the high failure rates of vascular reconstruction procedures. Local sustained drug delivery has shown promise in the prevention of restenosis. The drug release rate from mithramycin-loaded EVA matrices (0.1%) was evaluated, and their antirestenotic effect was studied in the rat carotid model and rabbit model of vascular grafts. The modulation of c-myc expression by mithramycin treatment was examined by immunohistochemistry in the rat carotid model. The proliferative response of injured rat arteries was studied by bromdeoxyuridine (BrdU) immunostaining. The impact of mithramycin treatment on vasomotor responses of the venous segments grafted into arterial circulation was studied ex vivo using vasoreactive compounds. Mithramycin was released exponentially from EVA matrices in PBS. Matrices co-formulated with PEG-4600 revealed enhanced release kinetics. The perivascular implantation of drug-loaded EVA-PEG matrices led to 50% reduction of neointimal formation, and reduced the c myc expression and BrdU labeling in comparison to control implants. Decreased sensitivity of mithramycin-treated grafts to serotonin-induced vasoconstriction was observed. Local perivascular mithramycin treatment limits the functional alteration caused by the grafting of venous segments in high-pressure arterial environment, and potently inhibits stenosis secondary to grafting and angioplasty injury. The antirestenotic effect is associated with reduced c-myc expression and with subsequent decrease in SMC proliferation. PMID- 11733086 TI - Different pharmacokinetics of nicotine following intravenous administration of nicotine base and nicotine hydrogen tartarate in rats. AB - Pharmacokinetics of nicotine was studied in rats following intravenous (i.v.) administration of nicotine base (NB) and nicotine hydrogen tartarate salt (NS) at a nicotine dose of 1 mg/kg. The area under the plasma concentration-time curve (AUC), mean residence time (MRT), systemic clearance (CL), distribution volume at steady state (V(ss)) and terminal plasma half-life (T(1/2,beta)) of nicotine were compared between NB and NS. Compared to NS, NB exhibited higher and sustained plasma nicotine levels, thereby yielding significantly (P<0.05) larger AUC (66.3 vs. 27.7 microg ml/min), MRT (165.7 vs. 58.3 min), T(1/2,beta) (144.2 vs. 51.4 min) and a lower CL (18.3 vs. 46.3 ml/min per kg). The V(ss) was comparable between the two compounds. The metabolic conversion to cotinine from NS was threefold larger than that from NB. The plasma protein binding and distribution to blood cells were comparable between the compounds. The apparent partition coefficient (APC) of NS decreased as a function of its concentration, while that of NB remained nearly constant. Particles of different mean sizes were observed for the 1% (w/v) aqueous solutions of NS (388.6 nm) and NB (123.8 nm). Different metabolism and/or elimination between NB and NS appear to be mainly responsible for their different pharmacokinetics. PMID- 11733087 TI - Surface hydrophobicity of particles is not necessarily the most important determinant in their in vivo disposition after intravenous administration in rats. AB - The in vivo disposition of polystyrene microsphere (MS) with the particle size of 50 nm (MS-50) and lecithin-coated MS-50 (LMS-50) after intravenous administration to rats was characterized. While a rapid elimination from the systemic circulation was observed for MS-50, much more prolonged circulating property was observed for LMS-50. In addition, this in vivo disposition property of LMS-50 was suggested to be ascribed to its lower affinity to the liver, which is the determining organ of the in vivo disposition of MS-50. The evaluation of surface hydrophobicity of MS-50 and LMS-50 in buffer solution revealed that the surface of MS-50 is more hydrophobic than that of LMS-50. However, LMS-50 was oppositely found to be more hydrophobic than that of MS-50 in rat serum. The profiles of serum proteins associated with MS-50 and LMS-50 were also examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that the amounts of some adsorbed proteins are greatly different between MS-50 and LMS-50. From these findings, it was suggested that the substantial difference in the in vivo disposition between MS-50 and LMS-50 would not be attributed to the difference in their surface hydrophobicity in the blood, but the difference in the type of serum proteins associated with them. PMID- 11733088 TI - Targeted PEG-based bioconjugates enhance the cellular uptake and transport of a HIV-1 TAT nonapeptide. AB - We previously described the enhanced cell uptake and transport of R.I-K(biotin) Tat9, a large ( approximately 1500 Da) peptidic inhibitor of HIV-1 Tat protein, via SMVT, the intestinal biotin transporter. The aim of the present study was to investigate the feasibility of targeting biotinylated PEG-based conjugates to SMVT in order to enhance cell uptake and transport of Tat9. The 29 kDa peptide loaded bioconjugate (PEG:(R.I-Cys-K(biotin)-Tat9)8) used in these studies contained eight copies of R.I-K(biotin)-Tat9 appended to PEG by means of a cysteine linkage. The absorptive transport of biotin-PEG-3400 (0.6-100 microM) and the bioconjugate (0.1-30 microM) was studied using Caco-2 cell monolayers. Inhibition of biotin-PEG-3400 by positive controls (biotin, biocytin, and desthiobiotin) was also determined. Uptake of these two compounds was also determined in CHO cells transfected with human SMVT (CHO/hSMVT) and control cells (CHO/pSPORT) over the concentration ranges of 0.05-12.5 microM and 0.003-30 microM, respectively. Nonbiotinylated forms of these two compounds, PEG-3350 and PEG:(R.I-Cys-K-Tat9)8, were used in the control studies. Biotin-PEG-3400 transport was found to be concentration-dependent and saturable in Caco-2 cells (K(m)=6.61 microM) and CHO/hSMVT cells (K(m)=1.26 microM). Transport/uptake was significantly inhibited by positive control substrates of SMVT. PEG:(R.I-Cys K(biotin)Tat9)8 also showed saturable transport kinetics in Caco-2 cells (K(m)=6.13 microM) and CHO/hSMVT cells (K(m)=8.19 microM). Maximal uptake in molar equivalents of R.I-Cys-K(biotin)Tat9 was 5.7 times greater using the conjugate versus the biotinylated peptide alone. Transport of the nonbiotinylated forms was significantly lower (P<0.001) in all cases. The present results demonstrate that biotin-PEG-3400 and PEG:(R.I-Cys-K(biotin)Tat9)8 interact with human SMVT to enhance the cellular uptake and transport of these larger molecules and that targeted bioconjugates may have potential for enhancing the cellular uptake and transport of small peptide therapeutic agents. PMID- 11733089 TI - A study of doxorubicin loading onto and release from sulfopropyl dextran ion exchange microspheres. AB - The objective of this study was to investigate various factors that influence doxorubicin (Dox) loading onto and release from sulfopropyl dextran ion-exchange microspheres (MS), and to evaluate the anticancer activity of the released drug in vitro. Dox was incorporated into the MS by incubating the MS with aqueous solutions of Dox at room temperature. The drug release was carried out at 37 degrees C in aqueous solutions containing NaCl with or without CaCl2. The kinetics of drug absorption and release, the amount of Dox released, and the stability of Dox after loading, freeze-drying, and release were determined by spectrophotometry. The cytotoxicity of Dox (the original drug or that released from MS) against murine EMT6 breast cancer cells was assessed using a clonogenic assay. An increase in the MS to drug ratio resulted in a higher absorption rate and a higher fraction of the drug extracted from the solution. The release rate and the equilibrium fraction of Dox released increased with a decrease in the initial amount of Dox loaded or an increase in the salt concentration. The addition of divalent ions (Ca2+) promoted drug release compared to NaCl alone. The percent loss of colony forming ability of the cells, a measure of cytotoxicity of the released Dox, was the same as parent Dox solutions, indicating that the drug bioactivity was fully preserved after the drug loading and release cycle. This work demonstrated that various drug release rates were achieved by varying the drug loading and that the MS-delivered Dox was effective against the cancer cells in vitro. PMID- 11733090 TI - Poly(amidoamine)-mediated intracytoplasmic delivery of ricin A-chain and gelonin. AB - Poly(amidoamine)s (PAAs) are water-soluble synthetic polymers designed to be biodegradable and biocompatible. Moreover, they display membrane disruptive properties in response to a decrease in pH. This attribute confers PAAs with endosomolytic properties in vitro and in vivo. A model system was developed to quantify their ability to promote the endosomal escape of macromolecules that may be interesting as therapeutic agents. Here, two PAAs (ISA 1 and 4) were incubated with B16F10 cells in vitro together with two non-permeant toxins: either ricin A chain (RTA) or gelonin. The relatively non-toxic PAAs ISA 1 and 4 (IC50>1.5 mg/ml) restored activity to the inherently inert toxins. The IC50 values for the ISA 1/RTA and ISA 1/gelonin combinations were 0.65+/-0.05 and 0.55+/-0.12 mg/ml, respectively. Similarly, when ISA 4 was incubated with a non-toxic combination of RTA and gelonin the IC50 value decreased to 0.57+/-0.03 and 0.43+/-0.26 mg/ml, respectively. In contrast, the neutral polymer dextran and the PAA ISA 22 were unable to mediate this effect. These observations suggest that specific PAA-toxin combinations warrant further development as novel therapeutics. PMID- 11733091 TI - N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers for targeted delivery of 8 aminoquinoline antileishmanial drugs. AB - A challenge to successful chemotherapy of visceral leishmaniasis is the dose limiting toxicity of antileishmanial agents. One approach to increase the efficacy and reduce the toxicity of these agents is to direct the drug to the phagolysosomes of the reticuloendothelial system (RES) where the leishmanial parasites reside. In this work a series of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-antileishmanial drug conjugates containing lysosomally degradable side chains and with or without sugar targeting moieties were synthesized, characterized and investigated for their in vivo efficacy in mice infected with Leishmania. An 8-aminoquinoline analog, namely 8-[(4-amino-1 methylbutyl)amino]-5-[3,4-dichlorophenoxy]-6-methoxy-4-methylquinoline (NPC1161) was used as a model antileishmanial agent. At 5 mg/kg body weight drug equivalent dose, all HPMA copolymer-drug conjugates which contained lysosomally degradable side chains showed significant in vivo antileishmanial activity (>99% inhibition), comparable to the activity of the free drug. At 2 mg dose, the same conjugates were significantly more effective (84-90% inhibition) than the free drug (67% inhibition). These results indicate the potential of lysosomotropic HPMA copolymers for the targeted delivery of antileishmanial compounds in the treatment of visceral leishmaniasis. PMID- 11733093 TI - Development of polymer film dosage forms of lidocaine for buccal administration. I. Penetration rate and release rate. AB - We examined the penetration rate of lidocaine (LC) through excised oral mucosa from hamster cheek pouch and the in vitro release rate of LC from film dosage forms with hydroxypropylcellulose (HPC) as a film base. Addition of glycyrrhizic acid (GL) to the HPC films increased the LC release rate almost GL-content dependently, while an optimum GL content was observed for the LC penetration rate. No LC penetration was observed from an acidic aqueous solution (pH 3.4) of LC, suggesting only unionized LC can substantially penetrate through the mucosa. A significant relationship between the penetration rate of LC and the release rate of unionized LC was found, suggesting that the in vitro dissolution study is a useful tool to predict the penetration rate taking the unionized drug fraction into consideration. PMID- 11733092 TI - An investigation of pulsatile release tablets with ethylcellulose and Eudragit L as film coating materials and cross-linked polyvinylpyrrolidone in the core tablets. AB - To develop new pulsatile release tablets, which can suppress drug release in stomach and release the drug rapidly after a predetermined lag time of about 3 h in intestine, the use of tablets with ethylcellulose/Eudragit L as a coating film and cross-linked polyvinylpyrrolidone in the core tablets was investigated. The release of diltiazem hydrochloride (DIL) as a model drug in the core tablets was investigated in vitro. The lag time (t10) was prolonged with an increase of the coating level, whereas the drug release rate was almost constant, irrespective of the coating level. The water-uptake study and electron microscope photographs suggested the mechanism of pulsatile release of drug. Pulsatile release tablets containing 60 mg DIL with 4.4 h of lag time (t10) in vitro were administrated to eight volunteers. The mean plasma concentration curves showed 4.9 h of lag time (tlag), 8.0 h of time to maximum concentration (tmax) and 3.1 h of time between tmax and tlag (t(psi)) in vivo. Relative bioavailability was 1.05 for pulsatile release tablets compared to conventional tablets. PMID- 11733094 TI - Sustained release of human growth hormone (hGH) from collagen film and evaluation of effect on wound healing in db/db mice. AB - Collagen films containing human growth hormone (hGH) were prepared and the release of hGH from these films and their effect on healing of full-thickness wounds in db/db mice were evaluated. The release profiles of hGH from the collagen films varied with composition and preparation conditions. The film prepared by air-drying of the mixture of hGH and collagen solution released hGH continuously over 3 days both in vitro and in vivo. By application of collagen film containing 3 mg of hGH twice at an interval of 6 days to wounds, area of wounds on day 21 was significantly reduced compared with that of non-treated wounds. Application of hGH alone at the same dose had no significant effect on wound healing. The maximum serum hGH concentration after single administration of the hGH collagen film was lower than that with hGH alone, and hGH persisted in serum over 3 days. These results suggest that hGH collagen film may be a useful topical formulation for the treatment of wounds. PMID- 11733095 TI - On the optimization of drug release from multi-laminated polymer matrix devices. AB - This work presents a systematic optimization framework to achieve desired release rates in drug delivery devices using multi-laminated layers. A simple mathematical model is used to describe the transient mass transfer between successive layers, laminated together to form matrices with different initial concentrations, drug diffusivities and thickness. First, an efficient analytical based optimization approach is investigated to define the optimal nonuniform initial drug distribution for constant diffusivity profile. The results obtained are in a good agreement with relevant work from the literature resorting to advanced optimal control techniques. Then, a formal dynamic optimization approach is employed, to systematically explore the synergistic benefits when all the available controllable parameters are simultaneously optimized, in order to achieve a drug release profile as close to a desired profile as possible for the entire period of operation. The optimization results lead to significantly improved constant release profiles. PMID- 11733096 TI - Enzyme loaded biodegradable microspheres in vitro ex vivo evaluation. AB - Prolidase is a naturally occurring enzyme involved in the final stage of protein catabolism. Deficient enzyme activity causes prolidase deficiency (PD), a rare autosomal recessive inherited disorder whose main manifestations are chronic, intractable ulcerations of the skin, particularly of lower limbs. Although several attempts have been made towards the treatment of this pathology, a cure for this disease has yet to be found. The purpose of this work is to evaluate the possibility of enzyme replacement therapy through prolidase microencapsulation in biodegradable microspheres. The poly(D,L-lactide-co-glycolide) (PLGA) prolidase loaded microparticulate systems have been prepared utilizing the w-o-w double emulsion solvent evaporation method. They have been characterized "in vitro" by morphological analysis, total protein content and an in vitro dissolution test of active protein. "Ex vivo" evaluation of prolidase activity from the microspheres has been performed on cellular extracts of cultured skin fibroblasts from healthy subjects (controls) and from patients affected by PD. The results reported in this work on prolidase from pig kidney (available on the market) demonstrate the positive role of microencapsulation as a process of enzymatic activity stabilization inside PLGA microspheres achieving both in vitro and ex vivo active enzyme release. This formulation can be proposed as a parenteral depot drug delivery system. PMID- 11733097 TI - Factors affecting protein release from alginate-chitosan coacervate microcapsules during production and gastric/intestinal simulation. AB - A series of experiments was performed to evaluate the influence of a number of physico-chemical factors on the diffusion of a model protein, bovine serum albumin (BSA), from dried chitosan-coated alginate microcapsules. Diffusion of BSA was quantified during the microcapsule manufacture processes (gelation, washing, rinsing) and during incubation in conditions simulating the pH encountered during the gastric (0.1 N HCl; pH 1.5) and intestinal (200 mM Tris HCl; pH 7.5) phases of digestion. Factors tested included alginate and chitosan concentration, calcium chloride (CaCl2) concentration in the gelation medium, loading rate, chitosan molecular mass and pH of the gelation medium. Microcapsule size and gelation time were altered in order to determine their effects on protein retention. Alginate and chitosan concentration significantly influenced BSA retention during microcapsule manufacture and acid incubation, as did calcium chloride concentration in the gelation medium (P<0.05). BSA retention during manufacture was not significantly altered by protein loading rate or pH of the encapsulation medium, however, protein retention during acid incubation decreased significantly with increasing protein loading rate and encapsulation medium pH (P<0.05). Microcapsules that were washed with acetone following manufacture demonstrated significantly increased protein retention during acid incubation (P<0.05). In microcapsules that had been acetone-dried to a point whereby their mass was reduced to 10% of that immediately following encapsulation, protein retention was over 80% following 24-h acid incubation vs. only 20% protein retention from non acetone-dried microcapsules. The presence of calcium in the neutral buffer medium significantly reduced BSA diffusion in a concentration dependent manner (P<0.05). PMID- 11733098 TI - Synthesis and characterization of poly(ethyl-2-cyanoacrylate) nanoparticles with a magnetic core. AB - A method is described to prepare composite colloidal nanoparticles, consisting of a magnetic core (magnetite) and a biodegradable polymeric shell (poly(ethyl-2 cyanoacrylate) or PE-2-CA). The method is based on the so-called anionic polymerization procedure, often used in the synthesis of PE-2-CA nanospheres designed for drug delivery. In the present work, the heterogeneous structure of the particles can confer both magnetic-field responsiveness and potential applicability as a drug carrier. In order to investigate to what extent this target is achieved, we compare the structure, chemical composition, and surface properties of the core/shell particles with those of both the nucleus and the coating material. This preliminary study shows that the synthetic new material displays an intermediate behavior between that of magnetite and PE-2-CA spheres. Thus, electrophoresis measurements as a function of pH and as a function of KNO3 concentration, show great similarity between the core/shell and pure polymer nanoparticles. A similar conclusion is reached when a surface thermodynamic study is performed on the three types of particles: the electron-donor component of the surface free energy of the solids is the quantity that appears to be most sensitive to the surface composition. The fact that PE-2-CA is close to being a non-polar material gives rise to a measurable decrease in the electron-donor component of the surface free energy of core/shell particles as compared to magnetite. PMID- 11733099 TI - Development and in vitro evaluation of a mucoadhesive vaginal delivery system for progesterone. AB - The purpose of the present study was to design a novel carrier system based on a mucoadhesive polymer exhibiting improved properties concerning drug delivery to the vaginal mucosa. This was reached by the covalent attachment of L-cysteine to commercially available polyacrylic acid (Carbopol 974P). Mediated by a carbodiimide, increasing amounts of L-cysteine were covalently linked to the polymer. The resulting thiolated polyacrylic acid conjugates (NaC974P-Cys) displayed between 24.8 and 45.8 micromol thiol groups per gram of polymer. Because of the formation of intra- and/or intermolecular disulfide bonds, the viscosity of an aqueous thiolated polymer gel (3%) increased about 50% at pH 7.0 within 1 h. In oscillatory rheological measurements, it was shown that this increase in viscosity is mainly due to the increase in elasticity. Tensile studies carried out on freshly excised cow vagina demonstrated a significant (P<0.05) increase in the total work of adhesion (TWA) compared to the unmodified polymer. An amount of 24.8 micromol thiol groups per gram of polymer resulted in a 1.45-fold increase in the TWA, whereas an amount of 45.8 micromol showed an even 2.28-fold increase. These improved mucoadhesive properties can be explained by the formation of disulfide bonds between the thiolated polymer and cysteine rich subdomaines of the mucus layer. The release rate of the model drug progesterone from tablets based on microcrystalline cellulose serving as the reference was approximately 1% per hour, whereas it was 0.58% per hour for the unmodified polymer (NaC974P) and 0.12% per hour for the thiolated polymer (NaC974P-Cys). Therefore, this thiolated polymer is a promising carrier for progesterone providing a prolonged residence time and a controlled drug release. PMID- 11733100 TI - Evaluation of a mucoadhesive tablet for ocular use. AB - The present study investigates the use of a polymer mixture containing Carbopol 974P and drum dried waxy maize starch to obtain prolonged drug release to the anterior eye segment. Two dosage forms with this composition are compared: a hydrated polymer dispersion and a minitablet. A model fluorescent tracer is used to study the ocular release and diffusion from the two dosage forms in humans. To evaluate the prolongation in the cornea/tearfilm compartment, the Apparent Fluorescein TurnOver (%/min) is calculated. The parameters Cmax, tmax, and C9h are used to characterize the pharmacokinetics of Na-fluorescein in the anterior chamber. Furthermore, the swelling behavior of the minitablet is evaluated macroscopically, while the degree of interaction with mucin is characterized by rheological measurements. Calculation of an acceptability score and a slug irritation potential is performed to evaluate user acceptability. In contrast to the hydrated dispersion, the minitablet significantly decreases the Apparent Fluorescein TurnOver (%/min) (P<0.05) and increases the apparent fluorescence in the anterior chamber 9 h after application of the preparation. Rheological data demonstrate the presence of elastic interactions between the polymer and mucin. The dry core of the minitablet becomes fully hydrated after approximately 2 h and is subsequently transformed into a highly concentrated gel. The acceptability of the minitablet is comparable to that of the polymer dispersion. Prolonging the release of Na-fluorescein to the anterior eye segment is only feasible with the dry preparation. PMID- 11733101 TI - Cationic solid-lipid nanoparticles can efficiently bind and transfect plasmid DNA. AB - The suitability of cationically modified solid-lipid nanoparticles (SLN) as a novel transfection agent was investigated. SLN were produced by hot homogenisation using either Compritol ATO 888 or paraffin as matrix lipid, a mixture of Tween 80 and Span 85 as tenside and either EQ1 (N,N-di-(beta steaorylethyl)-N,N-dimethylammonium chloride) or cetylpyridinium chloride as charge carrier. The resulting particles were approximately 100 nm in size and showed zeta potentials around +40 mV at pH 7.4. DNA binding was tested by agarose gel electrophoresis. The resulting SLN-DNA complexes were further characterised by AFM and zeta potential measurements. Only the SLN batch SII-13, composed of 4% Compritol, 4% Tween/Span and 1% EQ1, was able to form stable complexes with DNA. Typical complexes were 300 to 800 nm in size. Cytotoxicity and transfection efficiency was tested in vitro on Cos-1 cells. Cationic SLN produced by modification with EQ1 were well tolerated, with LD50 values >3 mg/ml in the LDH release assay and >0.6 mg/ml in the WST-1 assay. Further, SLN-DNA complexes containing between 10 and 200 weight equivalents of SII-13 (matrix lipid) efficiently transfected the galactosidase expression plasmid pCMVbeta in the absence and presence of the endosomolytic agent chloroquine. PMID- 11733102 TI - Effect of cholesteryl side chain and complexing with cholic acid on gene transfection by cationic poly(ethylene glycol) derivatives. AB - A new class of poly(ethylene glycol) derivative, Chol-PEG-A, having both cholesteryl- and amino-pendant groups (2.89 and 5.39 groups per polymer molecule, respectively) was synthesized. This amphiphilic PEG derivative forms a cationic polymer assembly in water. Chol-PEG-A expressed high transfection efficiency in the serum-free medium at relatively low amine/phosphate (N/P) ratios (1.3-1.5), whereas PEG-A, having only amino pendants without the cholesteryl side chain could transfect the cells only at a very high N/P ratio (83). The efficiency remarkably decreased by addition of fetal bovine serum (FBS) to the medium. Mixing of cholic acid to Chol-PEG-A gave a neutralized polyion complex, and the interaction with serum proteins was evidently suppressed. This Chol-PEG-A-cholic acid system showed higher gene expression even in the FBS-containing medium and as high a transfection efficiency as a Superfect. PMID- 11733104 TI - Mitochondrial transcription factor A (mtTFA) and diabetes. AB - Mitochondrial DNA (mtDNA) content decreased in an age-dependent manner and may be one of the causal factors in age-related type 2 diabetes. Mitochondrial transcription factor A (mtTFA), which provides the replication primer, plays a key role for the regulation of mtDNA replication and its level is proportional to mtDNA. Here, we studied on the regulatory mechanism of mtTFA expression and the factors affecting the transcriptional activity of the mtTFA promoter. The promoter of human mtTFA contains 67 CpG dinucleotides. When the plasmids bearing the mtTFA promoter (2378 bp) linked to luciferase were transiently transfected into HepG2 cells, in vitro methylation of NRF-1 site by HhaI methylase abolished the mtTFA promoter activity up to 90%, implying that the CpG methylation of NRF-1 site inactivate mtTFA promoter-driven transcriptional activity. Besides the promoter methylation, the exogenous hydrogen peroxide or glucose also modulates the promoter activity of mtTFA. The bacterially overexpressed mtTFA protein exhibits a strong binding affinity to circular DNA (perhaps to mtDNA in mitochondria in vivo) and the protection of the DNA from cleavage by a hydrogen peroxide attack. Taken all these results together, age-related alterations of oxidative stress may affect mtDNA replication via regulating mtTFA activity. Furthermore, a vicious cycle may be present between mtTFA protein level and oxidative stress in the sense of DNA damage. Further studies were necessary to prove the presence of methyl cytosine in the mtTFA promoter of either an aged or a diabetic person and the effect of oxidative stress on the mtTFA function and expression resulting in a change of the mtDNA copy number. PMID- 11733105 TI - Role of vitamin A in mitochondrial gene expression. AB - Diabetes-prone BHE/Cdb and Sprague-Dawley (SD) rats were studied with respect to mitochondrial (mt) function and mt gene expression. The BHE/Cdb rats carry mutations in the mt ATPase 6 gene that phenotype as decreased OXPHOS efficiency with subsequent development of impaired glucose tolerance. The base substitutions result in amino acid substitutions in the proton channel and this, in turn, affects the efficiency of energy capture in the ATP molecule. Feeding studies showed that BHE/Cdb rats required 10 times more vitamin E and three times more vitamin A in their diets than do normal SD rats. Vitamin A supplementation 'normalized' mt OXPHOS as well as increased the amount of ATPase subunit a protein in the mt compartment. Western blot analysis of retinoic acid receptors in the mitochondrial and nuclear compartments showed that these proteins were present in the mt compartment. The effect of the vitamin A supplementation plus the observation of retinoic acid receptors suggest that vitamin A functions to enhance the transcription of the ATPase 6 gene. Work with primary cultures of hepatocytes showed that not only does retinoic acid increase mitochondrial ATPase 6 gene expression but so too does the steroid hormone intermediate, dehydroepiandrosterone (DHEA). Triiodothyronine also plays a role in this process but not as an independent factor. Rather, this hormone potentiates the effects of retinoic acid and DHEA on ATPase gene expression. These results suggest that mt gene expression requires more than just the mt transcription factor A. More than likely the process requires a number of factors in much the same way as does nuclear gene expression. PMID- 11733106 TI - Oxidative stress markers in Korean subjects with insulin resistance syndrome. AB - Insulin resistance syndrome (IRS) is a cluster of prevalent conditions including glucose intolerance, hypertension and dyslipidemia, which commonly predispose to cardiovascular disease. However, the mechanism by which IRS is related with cardiovascular disease is not yet settled. Recently, it has been hypothesized that atherosclerosis is an inflammatory disease and that an increase in oxidative stress plays a key role in causing endothelial dysfunction associated with atherosclerosis. There has been, however, no study directly relating IRS with oxidative stress in human subjects. We measured various markers of oxidative stress among subjects who participated in a population-based epidemiological study performed in 1996. IRS was defined as non-diabetic subjects having more than two of three salient features of the syndrome (glucose intolerance, hypertriglyceridemia/low high density lipoprotein (HDL)-cholesterol and hypertension). The subjects with IRS (n=70) showed higher plasma malondialdehyde (MDA; 2.10+/-1.43 vs. 1.63+/-1.21 micromol/ml, P=0.009), homocysteine (16.32+/ 8.34 vs. 13.06+/-6.49 micromol/l, P=0.002) and ceruloplasmin concentrations (29.80+/-5.28 vs. 27.39+/-5.10 mg/dl, P=0.002) than control subjects (n=196). Plasma MDA concentration was positively correlated with waist-to-hip ratio (r=0.124, P=0.044), and with plasma triglyceride (TG; r=0.163, P=0.008), ferritin (r=0.200, P=0.002) and homocysteine concentrations (r=0.136, P=0.032). These results suggest that increase in oxidative stress may contribute to the development of cardiovascular disease in IRS. PMID- 11733107 TI - Prevalence and clinical characteristics of mitochondrial tRNAleu(UUR) nt 3243 A- >G and nt 3316 G-->A mutations in Chinese patients with type 2 diabetes. AB - Seven hundred and sixteen randomly selected, unrelated patients with type 2 diabetes were screened for mutations using a PCR-RFLP technique to assess the prevalence of the A-G mutation at position 3243 of the mitochondrial (mt) tRNAleu(UUR) gene in type 2 diabetes in the Chinese population. Three individuals with this mutation were identified, representing approximately 0.4% of the type 2 diabetes patients screened. Further screening of the first-degree relatives of these three patients identified another four affected carriers. In comparison with type 2 diabetic patients without the mutation, these seven carriers of the mt 3243 mutation had; (1) had an earlier onset of diabetes (38.0+/-10.1 vs. 53.4+/-10.0 year, P<0.001); (2) a lower body mass index (BMI) (19.5+/-2.0 vs. 24.9+/-10.9, P<0.0001); and (3) and lower post-challenge insulin levels (area under the curve of insulin levels during the OGTT, 2946+/-1647.2 vs. 7469+/ 6647.7, P<0.01). In addition, the same 716 patients with type 2 diabetes, as well as 181 controls with normal glucose tolerance, were screened for a newly described mt 3316 G-A mutation. This mutation was found in 16 patients with type 2 diabetes (2.2%) and five controls (2.7%). Therefore, the frequency of the mutation was not significantly different in the patients and controls. Moreover, the clinical characteristics such as the age of the onset of diabetes, the BMI, and insulin levels were not significantly different between the diabetic patients with the mt 3316 G-A mutation and those without. This shows that the mt 3316 G-A mutation is a polymorphism unrelated to diabetes. PMID- 11733108 TI - Molecular analysis of diabetes mellitus-associated A3243G mitochondrial DNA mutation in Taiwanese cases. AB - Investigation of the clinical manifestations of MELAS-specific A3243G mitochondrial DNA (mtDNA) point mutation has suggested that the A3243G mutation of mtDNA can cause certain subtypes of diabetes mellitus (DM) and contributes about 0.15% of the overall incidence of diabetes. However, a relationship between the diabetic syndrome and the proportion of mutant mtDNA in affected tissues remains unclear. In this article, we report the results of our investigation of 14 diabetic and 23 non-diabetic patients who had the A3243G mutant mtDNA. The proportions of mutant mtDNA in different tissues were noted to change variably and neither heteroplasmy of mutant mtDNA in various tissues nor the proportion of mutated mtDNA in a specific tissue showed a correlation with the clinical phenotype of DM. Generation of a diabetic syndrome was not predictable from either the content of mutant mtDNA in leukocytes, hair follicles, or in muscle tissues. Further study showed that muscle tissue has the highest proportion of mutant mtDNA followed by hair follicles and by blood cells. Moreover, we observed that as the patient's age increased, all tissue showed a declining proportion of mutant mtDNA. These findings suggest that age may play a role in the manifestation of diabetes in patients with A3243G mutation of mtDNA. PMID- 11733109 TI - Enhanced oxidative damage in human cells harboring A3243G mutation of mitochondrial DNA: implication of oxidative stress in the pathogenesis of mitochondrial diabetes. AB - Mitochondrial oxidative phosphorylation and the ATP production in pancreatic beta cells play significant roles in insulin secretion in response to glucose and other nutrients. An A to G mutation in the tRNA(Leu(UUR)) gene at nucleotide position (np) 3243 of mitochondrial DNA (mtDNA) has been observed in patients with MELAS syndrome and mitochondrial diabetes. Recently, some patients with mitochondrial diabetes associated with the A3243G mtDNA mutation were found to respond to coenzyme Q10 therapy. Thus, we investigated oxidative stress and peroxidative damage in a series of cybrids carrying either the wild-type adenine or the mutant-type guanine at np 3243 but having otherwise identical mtDNA sequence. The cybrids harboring >90% of the A3243G mutant mtDNA were found to have significantly lower oxygen consumption rate and electron transfer activities, and thereby had lower ATP/ADP ratios and declined energy charge. Importantly, the defective respiratory function elicited by the A3243G mtDNA mutation caused an increased oxidative stress as indicated by the decreased GSH/GSSG ratio and enhanced oxidative damage to lipids. Moreover, the cybrids harboring high proportions of the A3243G mtDNA mutation were found to be much more vulnerable to an exogenous oxidant, tert-butylhydroperoxide. We thus suggest that enhanced oxidative damage and elevated oxidative stress contribute to the decline of mitochondrial function and may be involved in the initiation and progression of the MELAS syndrome and mitochondrial diabetes. PMID- 11733110 TI - Method of proof and evidences for the concept that mitochondrial genome is a thrifty genome. AB - A hypothesis that mitochondrial genome itself is the thrifty genome has been advanced, as both its mutation, or quality, and amount in the peripheral blood, or quantity, are found to have close relationship to diabetes mellitus, or more specifically insulin resistance. Mere presence of relationship, however, does not establish cause-effect relation. Usually a genetic 'cause' is established, when a knockout of putative gene produces disease phenotype. However, absence of mitochondrial genome is incompatible with life. Here the argument advanced by Bradford Hill was applied to the proof of a hypothesis, as most of the data are from epidemiological observations; it is biologically plausible that both mtDNA quality and quantity are related to insulin resistance. The relationship had enough strength of association, dose-response relation, and consistency of relationship. Temporally association, decrease of mtDNA content preceding onset of diabetes, was also observed. Good hypothesis generates many predictions, which could be verified experimentally and which is biologically plausible. PMID- 11733111 TI - Endothelial dysfunction associated with oxidative stress in human. AB - The imbalance in pro- and anti-oxidant activity causes endothelial dysfunction, which precedes and exacerbates atherosclerosis. LDL-apheresis improves ischemic symptom in patients with hypercholesterolemia and regresses atherosclerosis. Whereas, patients who undergo hemodialysis experience accelerated atherosclerosis and premature death even though their cholesterol level is known to be normal. We investigated the association between oxidative modification and endothelial function during a single session of blood purification therapies, LDL-apheresis and hemodialysis. To examine the effect of a single session of LDL-apheresis on endothelial function in patients with hypercholesterolemia, we measured forearm blood flow (FBF) by strain gauge plethysmography before and after LDL-apheresis, while infusing acetylcholine (ACh) and sodium nitroprusside (SNP). The single session of LDL-apheresis reduced oxidized-LDL. Although ACh and SNP increased FBF dose-dependently before and after LDL-apheresis, ACh response was significantly augmented without changes in responses to SNP. The plasma level of oxidized LDL correlated with the degree of ACh-induced vasodilatation. To examine the effect of a single session of hemodialysis on endothelial function in patients with end stage renal disease, we estimated flow-mediated vasodilation (FMD) during reactive hyperemia using a high-resolution ultrasonography, before and after a single session of hemodialysis. We also investigated the effect of anti-oxidative modification during hemodialysis using an a-tocopherol acetate-coated or a non coated cellulose dialyzer. Non-specific endothelium independent vasodilation was measured after sublingual glyceryltrinitrate spray (GTN; 0.3 mg) administration in each session. A single session of hemodialysis increased plasma level of oxidized LDL. Although hemodialysis treatment did not affect GTN-induced vasodilation, a single session of hemodialysis impaired the endothelium-dependent vasodilation. Treatment with vitamin E-coated-dialyzer restored endothelium dependent vasodilation, which was accompanied by decrease in oxidized LDL. Thus, endothelial function was impaired by acute increase in oxidative stress and was restored by anti-oxidant therapy. Our results suggest the importance of controlling oxidative stress in addition to classical risk factors to preserve human endothelial function. PMID- 11733112 TI - Oxidative stress and vascular aging. AB - In attempt to meet tissue demands for proper blood flow, the vasculature alters its structure, simultaneously responding to both physical and chemical stresses. Substantial information has emerged in this field of study, particularly concerning the roles of the endothelium and smooth muscle cells in relation to signaling pathways for mechanotransduction. As a first line of defense upon exposure to various stressors, the endothelium and smooth muscle cells respond with adaptive cellular modifications. One prime example of these modifications is the cellular response to oxidative stress as evidenced by accumulated data. A recent proposal of the inflammatory hypothesis of vascular aging emphasized that stress-induced vascular aging may be the primary event that underlies the general aging phenomenon of systemic dysfunction. PMID- 11733114 TI - Polymeric materials for advanced drug delivery. PMID- 11733113 TI - Effect of estrogen on endothelial dysfunction in postmenopausal women with diabetes. AB - BACKGROUND: Treatment with estrogen causes increased release of nitric oxide and this appears to be an important mechanism involved in the cardioprotective effect of estrogen. Previous studies have clearly shown that estrogen has an acute vasodilatory effect. Recently, abnormality of flow mediated endothelial dilation, which is defined as an endothelial dysfunction, has been proposed as an early manifestation of atherogenesis. However, its effect in type 2 diabetes is unknown. Stimulus-provoked endothelium dependent dilatation, using high resolution external vascular ultrasound, is a useful method in the measurement of endothelial function. The main advantages of this method are that it is completely non-invasive, accurate and reproducible. This classical method for the measurement of endothelial dysfunction, utilizing non-invasive, high-resolution ultrasonography, has some limitations especially in terms of reproducibility. To improve the accuracy and reproducibility of this method, measurements made for this study involved the use of an additional attachment. This device was an ultrasound probe that was attached to the arm of patient. In addition, this study evaluated not only a change in diameter in terms of flow mediated vasodilation, but also initial and peak response times during this test. The aims of this study are: (1) to evaluate the flow mediated peak vasoreactivity (FMD), the lag time from base line to initiation of vasodilation (IRT) and the peak response of endothelial dependent vasodilation in a control group and in young diabetic patients; and (2) to evaluate the effect of estrogen treatment on flow mediated vasodilation in postmenopausal women with diabetes. METHOD: Measurements were taken for flow mediated vasodilation (endothelial dependent vasodilation: FMD), endothelial independent vasodilation (EID) and lag time from base line to initial reaction of FMD. The aforementioned non-invasive method and attachment were used. Subjects for the experiment were 12 young people with diabetes (six male and six female, mean age 26.3) and 12 age-matched healthy controls (six male and six female, mean age 25.6). In addition, measurements were taken for FMD, EID and lag time in the brachial artery before and after estrogen supplementation (Premarin 0.625 mg for 7 days). Subjects for the experiment were 16 normal postmenopausal women and 18 age-matched postmenopausal women with type 2 diabetes. RESULT: There is no significant difference in BMI (body mass index), mean age, or blood pressure between the control group and the young diabetic group. There were no differences in age, total and LDL cholesterol levels or body mass index between the groups of postmenopausal women (P<0.05). However, the HDL cholesterol level was found to be significantly lower in postmenopausal women with diabetes than in normal postmenopausal women (respectively, a mean+/-SD; 38.95+/-11.96 mg/dl vs. 52.20+/-9.55 mg/dl, P<0.05). The FMD of young diabetic patients is slightly lower than that of age-matched young healthy controls (healthy control: 8.9+/-2.7%, young diabetic patients: 6.3+/-2.1%, P<0.05). There was no difference in endothelial independent vasodilation between the control group and the young diabetic group. The IRT of FMD was significantly shorter in the healthy control group than in the young diabetic group (healthy control: 20.4+/-2.8 s, diabetic group: 29.5+/-5.4 s, P<0.0001). In contrast, the IRT of EID was no different between the two groups. The IRT of FMD showed a significant negative correlation with FMD (r=-0.61, P<0.01) and a significant negative correlation with high density lipoprotein (HDL) cholesterol (r=-0.68, P<0.0001). The basal endothelium dependent vascular reactivity was significantly decreased in postmenopausal women with diabetes compared with normal postmenopausal women (8.0+/-3.9 vs. 13.7+/ 6.2%, P<0.05). Estrogen supplementation increased endothelium dependent vasodilation not only in postmenopausal women with diabetes (from 8.0+/-3.9 to 15.1+/-4.0%, P<0.05) but also in normal postmenopausal women (from 13.7+/-6.2 to 20.1+/-4.7%, P<0.05). In contrast, the responses to sublingual nitroglycerin were comparable between postmenopausal women with diabetes (from 21.1+/-6.0 to 22.1+/ 4.1%, P>0.05) and normal postmenopausal women (from 25.8+/-7.8 to 25.2+/-4.5%, P>0.05), both before and after estrogen supplementation. CONCLUSION: These findings indicate that in addition to the percentage change in FMD, lag time from base line to initial response time (IRT) of FMD is a good indicator for evaluation of endothelial dysfunction. Also, this new parameter may be a more sensitive parameter for differentiation between the diseased state and the normal state of the endothelium than percentage changes in FMD. In terms of postmenopausal women, endothelial dysfunction was prominent in women with diabetes and was significantly improved by estrogen but not reversed. These results suggest that other factors in addition to estrogen deficiency play a role in endothelial dysfunction in postmenopausal women with diabetes mellitus. PMID- 11733115 TI - Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoate. AB - Polyhydroxyalkanoates (PHAs) comprise a large class of polyesters that are synthesized by many bacteria as an intracellular carbon and energy compound. Analysis of isolated PHAs reveal interesting properties such as biodegradability and biocompatibility. Research was focused only recently on the application of PHA in implants, scaffolds in tissue engineering, or as drug carriers. Such applications require that PHA be produced at a constant and reproducible quality. To date this can be achieved best through bacterial production in continuous culture where growth conditions are kept constant (chemostat). Recently, it was found that PHA producing bacteria are able to grow simultaneously limited by carbon and nitrogen substrates. Thus, it became possible to produce PHA at high yields on toxic substrate and also control its composition accurately (tailor made synthesis). Finally, applications of PHA in medicine are discussed. PMID- 11733116 TI - Biodegradable block copolymers. AB - Recently, block copolymers have got tremendous impetus on the ongoing research in the area of drug delivery technology, due to their capability to provide a biomaterial having a broad range of amphiphilic characteristics, as well as targeting the drugs to specific site. This article is an attempt to review applications of block copolymers in surface modification, drug targeting, nano and microparticles, hydrogels, micelles etc. The physicochemical properties of block copolymers and various synthetic routes for block copolymers are also discussed. PMID- 11733117 TI - Therapeutic applications of viscous and injectable poly(ortho esters). AB - Poly(ortho esters) (POE) are hydrophobic and bioerodible polymers that have been investigated for pharmaceutical use since the early 1970s. Among the four described generations of POE, the third (POE III) and fourth (POE IV) are promising viscous and injectable materials which have been investigated in numerous biomedical applications. POE III has been extensively studied for ophthalmic drug delivery, it presents an excellent biocompatibility and is currently being investigated as a vehicle for sustained drug delivery to treat diseases of the posterior segment of the eye. POE IV is distinguishable by a highly reproducible and controlled synthesis, a higher hydrophobicity, and an excellent biocompatibility. It is currently under development for a variety of applications, such as ocular delivery, periodontal disease treatment and applications in veterinary medicine. This review will also focus on new perspectives for this promising family of polymers, such as guided tissue regeneration, treatment of osteoarthritis, as well as peptide and protein delivery. PMID- 11733118 TI - Histidine-rich peptides and polymers for nucleic acids delivery. AB - Nucleic acids transfer into mammalian cells requires devices to improve their escape from endocytic vesicles where they are mainly confined following cellular uptake. In this review, we describe histidine-rich molecules that enable the transfer of plasmid and oligonucleotides (ODN) in human and non-human cultured cells. An histidine-rich peptide which permeabilizes biological membrane at pH 6.4, favored the transfection mediated by lactosylated polylysine/pDNA complexes. Histidylated polylysine forms cationic particles of 100 nm with a plasmid and yielded a transfection of 3-4.5 orders of magnitude higher than polylysine. The biological activity of antisense ODN was increased more than 20-fold when it was complexed with highly histidylated oligolysine into small cationic spherical particles of 35 nm. Evidence that imidazole protonation mediates the effect of these molecules in endosomes are provided. We also describe a disulfide containing polylysine conjugate capable of mediating DNA unpackaging in a reductive medium and to increase the transfection efficiency. Overall, these molecules constitute interesting devices for developing non-viral gene delivery systems. PMID- 11733119 TI - Advanced drug delivery devices via self-assembly of amphiphilic block copolymers. AB - Amphiphilic block copolymers are well established as building blocks for the preparation of micellar drug carriers. Over the past decade, the effectiveness of such self-assembled drug delivery devices has been demonstrated numerous times. This review will discuss two approaches that can be used to further improve the effectiveness of amphiphilic block copolymer-based drug delivery systems. The first approach involves the chemical modification of the block copolymer building blocks. Several examples will be discussed of amphiphilic block copolymers modified with crosslinkable groups in order to increase the stability of the micellar drug carriers, or of block copolymers containing specific ligands that could ultimately allow targeted drug delivery. The second approach to improve the performance of micellar drug carriers is the addition of auxiliary agents. To illustrate this approach, the feasibility of channel proteins and metal (nano)particles to improve temporal control over the drug release process is discussed. PMID- 11733120 TI - Responsive hydrophobically associating polymers: a review of structure and properties. AB - Responsive hydrophobically associating polymers can in many ways be considered to be analogous to proteins in their ability to form compact molecules with a defined secondary structure, and hence, functionality. These molecules are characterized by the presence of alternating charged and hydrophobic groups. The balance between charge repulsion and hydrophobic interactions is sensitive to environmental pH and therefore changes in pH produce controllable conformational changes. The change from a charged extended chain to a collapsed uncharged coil structure is sometimes referred to as hypercoiling behaviour and enables the polymer to act as a simple switch between an 'on' and 'off' state. The purpose of this review is to illustrate the structure and behaviour of polymers that exhibit hypercoiling behaviour and to highlight their potential pharmaceutical applications, which in terms of drug delivery is likely to be related to their surface behaviour and solubilizing activity. PMID- 11733121 TI - Combinatorial chemistry and biomedical polymer development. AB - Polymers are ubiquitous components of products manufactured for medical and pharmaceutical applications. Widely used commodity polymers were the first polymers to be utilised in biomedical applications. These polymers were not developed with biocompatibility established at the onset and many speciality polymers have been developed in recent years to begin to meet the multifaceted demands for medical development, the optimisation of structure-property correlations and ultimately, clinical use. In the broader area of materials research, combinatorial or high throughput strategies used for drug development are recognised to have potential for discovery and process development. Much of the application of combinatorial chemistry in drugs research has been dependent on the use of polymeric reagents, substrates and supports. The chemistry of the reactions on polymers in solid and liquid phases have also played a major role in combinatorial drugs research. There is considerable interest in combinatorial materials research and this review outlines how this research may be applied for biomedical polymer development. PMID- 11733123 TI - Biomechanical examination of a commonly used measure of spasticity. AB - BACKGROUND: An increase in the prevalence of neurological disability puts pressure on service providers to restrict costs associated with rehabilitation. Spasticity is an important neurological impairment for which many novel and expensive treatment options now exist. The antispastic effects of these techniques remain unexplored due to a paucity of valid outcome measures. AIM: To develop a biomechanical measure of resistance to passive movement, which could be used in routine clinical practice, and to examine the validity of the modified Ashworth scale. STUDY DESIGN: Repeated measure cross-section study on 16 subjects who had a unilateral stroke one-week previously and had no elbow contractures. OUTCOME MEASURES: Simultaneous measurement of resistance to passive movement using a custom built measuring device and the modified Ashworth scale. Passive range of movement and velocity were also measured. The "catch", a phenomenon associated with the modified Ashworth scale, was identified by the assessor using a horizontal visual analogue scale and biomechanically quantified using the residual calculated from a linear regression technique. RESULTS: Half the study population had a modified Ashworth score greater than zero. The association between the two measures was poor (kappa=0.366). The speed and range of passive movement were greater in subjects with modified Ashworth score "0" (P<0.05). Resistance to passive movement was higher in the impaired arm (P<0.05) and tended to decrease with repeated measures and increasing speeds. CONCLUSIONS: A device to measure resistance to passive movement at the elbow was developed. The modified Ashworth scale may not provide a valid measure of spasticity but a measure of resistance to passive movement in an acute stroke population. RELEVANCE: Spasticity is an important neurological impairment for which many novel and expensive treatment options are being made available. There is a paucity of clinically usable outcomes to measure spasticity. A device to measure resistance to passive movement at the elbow, which was more reliable than the modified Ashworth scale was developed. This device may provide a much needed objective clinical measure to evaluate the efficacy of antispasticity treatment. PMID- 11733124 TI - Dynamic loads on the upper extremities during two different floor cleaning methods. AB - BACKGROUND: Biomechanical models have been used frequently to estimate the load on the low back and the lower extremities during occupational work tasks. The shoulder region has received much less attention although this area is a common site of work-related musculoskeletal disorders. OBJECTIVE: The aim of the present study was to evaluate the mechanical loading of the upper extremities during floor cleaning performed with different techniques. DESIGN: A comparative study of six subjects performing floor cleaning with two different techniques (mopping and scrubbing) in a laboratory setting. METHODS: A "force handle" equipped with two force dynamometers was used for continuous measurement of three-dimensional forces on each hand, separately, during this two handed asymmetrical task. A link segment model of the upper extremities was used to calculate the mechanical loading of the elbows and shoulders. Electromyograms were recorded from six shoulder muscles. RESULTS: Despite differences in movement patterns between mopping and scrubbing, only small differences were found in the moments of force. In accordance similar levels of electromyograms were found. CONCLUSION: In preventive strategies, changing the cleaning tool may not be sufficient to change the load on the shoulder muscles. RELEVANCE: Combined three-dimensional video recording and force measurements on the hands during a two-handed asymmetric work tasks allow calculation of the mechanical load on the elbow and shoulder. Thus it is possible to evaluate physiologically, new equipment designed to prevent work related musculoskeletal disorders. Before introducing such equipment in large scale interventions it may be evaluated if the changes in force requirements are sufficient to predict prevention of musculoskeletal disorders. PMID- 11733125 TI - The anatomic features of the radial head and their implication for prosthesis design. AB - OBJECTIVE: To establish the anatomical features of the radial head of an average normal human being and to verify the hypothesis that no significant difference exists between the geometry of the left and right normal radial heads. DESIGN: 17 proximal ends of the radius from the left and right forearms of fresh male (average age 50 and range 40-70) cadavers were measured. BACKGROUND: A reconstruction of anatomical features of the normal bone is important for prosthesis design. METHODS: A morphologic study of the radial head was performed using a co-ordinate measuring machine integrated with a computer aided design system. For comparative purposes, a statistical analysis including linear regression and correlation was performed. RESULTS: The maximum diameter (mean 23.36 mm (SD, 1.14 mm)) and height (mean 10.14 mm (SD, 1.38 mm)) of the radial head as well as the depth (mean 1.92 mm (SD, 0.32 mm)) and maximum radius (mean 20.27 mm (SD, 4.61 mm)) of the concave articulate surface are the most important anatomical features, which should be implicated in prosthesis design. The inclinations mean 2.50 degrees (SD, 0.41 degrees ) and mean 9.50 degrees (SD, 0.52 degrees ) and shift (mean 1.71 mm (SD, 0.35 mm)) of the radial head relative to its neck should also be taken into account in prosthetic design. CONCLUSIONS: The results of the study showed that "left is equal to right" (no significant differences between sides were obtained, for probability value P>0.05). RELEVANCE: This paper describes a morphological study of the proximal radius. The results can be used to reconstruct the geometry of the injured radial head based on the obtained geometric features of the contra-lateral side. These results can be also used to design radial head prosthesis. PMID- 11733126 TI - Effect of elbow flexion on upper extremity impact forces during a fall. AB - OBJECTIVE: The overall objectives are to develop a biomechanical model for a simulated fall with outstretched hand. DESIGN: Cross-sectional study involving young healthy volunteers in a university research laboratory setting. BACKGROUND: Little is known about the factors which influence fracture risk during a fall on outstretched hand. METHODS: A group of 11 male subjects volunteered for this investigation. A set of eight reflective markers was placed bilaterally on selected anatomic landmarks. Subjects were suspended with both elbows extended and wrists dorsiflexed, preparing to impact the ground and force plates from two different fall heights: 3 and 6 cm. Two different postures for the elbows were employed. In the elbow extension experiment, the elbows were extended at all times. In the elbow flexion experiment, the elbows were extended at impact, but then flexed immediately, as though in the initial downward phase of a push-up exercise. RESULT: Increasing the fall height significantly increased the upper extremity axial forces by 10% and 5%. No significantly different differences were found in the axial forces applied to the wrist, elbow or shoulder between the elbow flexion and elbow extension trials, but the elbow mediolateral shear force was 68% larger (P=0.002) in the extension trials. CONCLUSIONS: Performing an elbow flexion movement at impact reduces the first peak impact force value and postpones the maximum peak value. Although changing the fall arrest strategy from elbow extension to elbow flexion did not affect the peak impact force on the hand, it did require substantially greater elbow and shoulder muscle strengths. RELEVANCE: This paper yields insights into how the physical demands of arresting mild falls may relate to upper extremity muscle capacity, joint dislocation and bony fracture. PMID- 11733127 TI - A minimally invasive method for the determination of force in the interosseous ligament. AB - OBJECTIVE: The objective was to develop and utilize a minimally invasive testing system to determine the force in the interosseous ligament under axial compressive loads across the range of motion of the human forearm. DESIGN: Eleven fresh frozen human cadaveric forearms were used (51-72 years). BACKGROUND: Current studies investigating interosseous ligament forces altered the structure of the forearm by implanting load cells into the radius and ulna. This may affect load transfer through the forearm. Little information was available on interosseous ligament function over the entire flexion range of the elbow. METHODS: A robotic joint testing system was used to apply a 100 N compressive load to the forearm and measure the resulting displacement. Each forearm was tested with no disruption of the bones and soft tissues of the forearm. The principle of superposition was used to calculate the forces in the interosseous ligament and was indirectly validated using fluoroscopy. RESULTS: The force in the interosseous ligament ranged from a minimum of 8 N in neutral forearm rotation at full extension to a maximum of 43 N in supination at 30 degrees of flexion. The largest force was found in supination at all flexion angles. CONCLUSIONS: The interosseous ligament is an important structure in the stability of the forearm. The force in the interosseous ligament depends on the elbow flexion angle and forearm rotation. RELEVANCE: This study suggests that radial head fractures are best treated with the forearm in supination, since the interosseous ligament takes the largest load in this position. Complex injuries which have a poor prognosis, may require interosseous ligament reconstruction to improve clinical outcomes. PMID- 11733128 TI - Heel pad stiffness in runners with plantar heel pain. AB - OBJECTIVE: To evaluate significant differences in heel pad stiffness within a cohort of runners with diagnosed plantar heel pain and to explore the clinical importance of maximum heel pad stiffness values. DESIGN: A cross-sectional design was used to quantify the heel pad stiffness of 166 runners with 33 diagnosed with plantar heel pain. BACKGROUND: Palpation is still widely used to evaluate heel pad stiffness subjectively in everyday clinical practice. However, there is limited quantifiable data pertaining to heel pad stiffness measurements in runners and those with heel pain. METHODS: A portable hand-held device measured force applied by a metal probe, and its displacement into the plantar surface of the heel pad. Non-linear modelling allowed curve coefficients b0 and b1 to be evaluated and was described by an exponential function using a non-linear regression equation. Exploratory analysis was used to describe a single-point approximation for clinical use. RESULTS: An independent t-test demonstrated a statistically significant difference between the curve coefficient b1 (p<0.05). No significant difference was found for coefficient b0 between the plantar heel pain group and the non-plantar heel pain group (p>0.05). Exploratory analysis demonstrated maximum mean stiffness of 3.22 N/mm for the non-plantar heel pain group and 2.87 N/mm for the plantar heel pain-group, an 11% mean difference. CONCLUSION: The results suggested that heel pad stiffness may be associated with plantar heel pain subjects. RELEVANCE: Heel pad stiffness measurements may give a better insight into the mechanical properties of the heel pad in subjects with plantar heel pain. PMID- 11733129 TI - Lower extremity kinetics during stair ambulation in patients with and without patellofemoral pain. AB - OBJECTIVE: To compare lower extremity kinetics during stair ascent and descent in subjects with and without patellofemoral pain. DESIGN: A cross-sectional study utilizing a control group. BACKGROUND: The patellofemoral joint reaction force (the resultant force between the quadriceps muscle force and patellar ligament force) increases with quadriceps force and knee flexion angle. Consequently, patients with patellofemoral pain may employ compensatory strategies to minimize pain and reduce patellofemoral joint reaction forces during activity. METHODS: 10 individuals with a diagnosis of patellofemoral pain and 10 individuals without pain participated. Subject groups were matched on sex, age, height, and body mass. Anthropometric data, three dimensional kinematics, and ground reaction forces were used to calculate lower extremity sagittal plane moments (inverse dynamics) while subjects ascended and descended stairs at a self-selected pace. Differences in kinetic variables between groups were assessed using 2x2 (group x stair condition) analysis of variance. RESULTS: Subjects with patellofemoral pain had decreased peak knee extensor moments during stair ascent and descent. There were no group differences in peak hip, ankle, or support moments, however, subjects with patellofemoral pain had decreased cadence (descent) compared to controls. CONCLUSION: Subjects with patellofemoral pain had reduced peak knee extensor moments, suggesting that quadriceps avoidance was employed to reduce patellofemoral joint reaction forces. The lack of group differences in peak moments at the hip and ankle suggests that secondary compensation did not occur exclusively at the hip or ankle in this group of subjects with patellofemoral pain. RELEVANCE STATEMENT: Because stair ambulation is often used to evaluate the reproducibility of symptoms and to identify abnormal movement patterns indicative of patellofemoral pain, knowledge of lower extremity mechanics during stair negotiation is necessary to better characterize compensatory behavior in this population. PMID- 11733130 TI - Preliminary comparison of the rupture of human and rabbit anterior cruciate ligaments. AB - OBJECTIVE: This study was aimed at examining ruptures of the human anterior cruciate ligaments by scanning electron microscopy and video imaging and comparing the appearance of the rupture surfaces with those from rabbit anterior cruciate ligaments. DESIGN: The specimens were tested to failure as femur anterior cruciate ligament-tibia complexes using an Instron 8511 materials testing machine. BACKGROUND: Rupture of the anterior cruciate ligament is a major clinical problem, leading to instability of the knee joint. Due to the frequency and potential severity of the injuries, a need still exists for information on the biomechanical properties of ligaments under loading conditions, which occur at the time of trauma. METHODS: Four human femur-anterior cruciate ligament-tibia complexes were loaded to failure at a displacement rate of 0.008 m/s. Video recordings of the tests were used to study the progress of the ruptures and to compare the modes of failure of the ligaments. Scanning electron microscopy was employed to study the appearance of collagen fibres at the rupture surfaces. RESULTS: The modes of failure of the rabbit anterior cruciate ligament and appearance of the rupture surfaces were similar to those of the human anterior cruciate ligaments. CONCLUSION: The rabbit anterior cruciate ligament provides a model for investigating failure of the human ligament during trauma. RELEVANCE: The results will be of significance since most studies on ligaments are carried out on animal models with the intention of applying the deductions from the results to human ligaments. Examining the appearance of collagen fibres at these surfaces may help us to understand more about what actually happens during and after ligament rupture. PMID- 11733131 TI - A transducer to measure isometric elbow moments. AB - OBJECTIVE: The purpose of this study was to design and implement a transducer to measure accurately the isometric elbow moments produced by individuals with tetraplegia. DESIGN: The device needed to be insensitive to off-axis moments and proximal joint motions and be capable of being used over a wide range of elbow and shoulder positions in an outpatient clinic setting. BACKGROUND: Measurement of the smaller isometric moments produced by individuals with tetraplegia is especially sensitive to the errors that can be introduced by inaccurate lever arm determination, off-axis loads, and proximal joint motions. Devices traditionally utilized for quantifying isometric strength are difficult to implement for the spinal cord injured population. METHODS: The elbow moment transducer consists of two four-bar parallelogram linkages joined by a lockable pivot. Strain gauges mounted on one beam of the parallelogram produce an output proportional to the elbow moment. RESULTS: Calibration of the device indicates that it accurately quantifies isometric elbow moments over a range that is appropriate for evaluating elbow extension strength in individuals with tetraplegia. CONCLUSIONS: A device was developed and implemented that accurately quantifies isometric elbow moments over a range that is appropriate for evaluating elbow extension strength in individuals with tetraplegia. RELEVANCE: The ability to quantitatively evaluate elbow strength in persons with tetraplegia is useful for understanding and improving the clinical outcomes of rehabilitative interventions that involve the elbow. PMID- 11733132 TI - Integration of plantar soft tissue stiffness measurements in routine MRI of the diabetic foot. AB - OBJECTIVE: A method of measuring mechanical properties of plantar soft tissues in vivo by integrating a photoelastic technique into an MRI system is presented. BACKGROUND: Plantar tissue stiffening in the diabetic foot is associated with development of ulcers at the highest-pressure sites. MRI is used as an effective means of demonstrating tissue damage caused by diabetic ulceration. DESIGN: The new measurement method uses the indentation of a sphere into the plantar tissue, which is simultaneously scanned by MRI. The indentation force is obtained using the photoelastic technique while tissue deflection is measured on the MRI scans, to yield a characteristic load-displacement curve. METHODS: Applicability of the proposed method in characterizing the plantar tissue stiffness and indentation patterns of two young normal subjects and two older diabetic patients was tested. RESULTS: Significantly higher effective shear and elastic moduli were measured for the diabetic plantar tissue compared with that of normal. For the diabetic patients, local stiffness in proximity of the 1st metatarsal head was substantially larger than in other sites, and this could be associated with initial tissue ulceration at this location. RELEVANCE: The present method seems applicable as a practical means of identifying regions in the diabetic plantar tissue that are vulnerable to ulceration, and can be easily integrated into routine MRI scans for assessing tissue damage in these patients. PMID- 11733133 TI - Can interventional MRI provide an insight into the mechanics of a posterior anterior mobilisation? AB - OBJECTIVES: A pilot study aimed to investigate if interventional MRI could detect changes in the spine and surrounding soft tissues as a result of a posteroanterior mobilisation, and to determine if such an effect was specific to the level of application. DESIGN: A prospective pilot study to investigate the kinematics of cervical spine mobilisation. BACKGROUND: Posteroanterior mobilisation is a manual physiotherapy technique that is commonly used as an examination tool and a form of conservative treatment. The efficacy of this technique is controversial and little is known regarding the underlying mechanisms. METHODS: Five control subjects were recruited and written informed consent obtained. Subjects were scanned, using a General Electric Signa SP10 Open Interventional MRI scanner, in the prone position with their necks in either a neutral or flexed position. In each position a posteroanterior mobilisation to the 2nd and 6th cervical vertebrae was performed. Sagittal images of the spine were obtained prior to, during and following the mobilisation. RESULTS: Clear images of vertebral position could be obtained during the application of a posteroanterior mobilisation. Measurements of intervertebral angulation and translation demonstrated little if any motion during the application of a posteroanterior mobilisation, in both a flexed and neutral position. Changes were seen in terms of soft tissue compression and in some instances overall angulation of the cervical spine. CONCLUSIONS: Interventional MRI can produce images of posteroanterior mobilisation. Dynamic imaging of the procedure produced images of unsuitable quality for analysis. The quasi-static images of suggested that the application of a posteroanterior mobilisation to the cervical spine produces minimal if any intervertebral motion, but significant soft tissue deformation of the overlying structures. RELEVANCE STATEMENT: Posteroanterior mobilisation is frequently used in the assessment and management of spinal problems, but the efficacy of the technique remains controversial. This is hampered by the fact that little is known about the mechanisms involved during such a procedure. PMID- 11733134 TI - ACL research retreat: the gender bias. April 6-7, 2001. Meeting report and abstracts. PMID- 11733135 TI - The SMN genes are subject to transcriptional regulation during cellular differentiation. AB - Proximal spinal muscular atrophy (SMA) is an autosomal recessive disease characterized by degeneration of alpha-motor neurons and muscular atrophy. The causal survival motor neuron (SMN) gene maps to a complex region of chromosome 5q13 harbouring an inverted duplication. Thus, there are two SMN genes, SMN1 and SMN2, but SMN1-deficiency alone causes SMA. In this study we demonstrate, for the first time, down-regulation of SMN promoter activity during cellular differentiation. Specifically, the minimal SMN promoter is four times more active in undifferentiated embryonal carcinoma P19 cells compared to cells treated with retinoic acid (RA) to initiate neuronal differentiation. This effect is mediated by sequences contained within the minimal core promoter that we have confined to the 257 nucleotides upstream of exon 1. We have identified seven regions that are highly conserved between the mouse and human SMN core promoters and this region contains the consensus sequence for a number of transcription factors. Most notably, AhR, HNF-3 and N-Oct3 have already been shown to respond to RA treatment of EC cells, while E47, HNF-3, MAZ, N-Oct3 and Pit-1a have been implicated in embryonic, muscle or neural development. In addition, we have mapped two strong transcription initiation sites upstream of SMN exon 1. The novel -79 site identified in this study is preferentially utilized during human foetal development. Furthermore, analysis of RNA from SMA patients with deletions of the entire SMN1 gene or chimpanzees that lack SMN2 suggests that the level of transcription initiation at these sites may be different for the SMN1 and SMN2 genes. Taken together, this work provides the first demonstration of transcriptional regulation of these genes during cellular differentiation and development. Deciphering the underlying mechanisms responsible for regulating SMN transcription may provide important clues towards enhancing SMN2 gene expression, one target for the treatment of SMA. PMID- 11733136 TI - The murine Wilms tumor suppressor gene (wt1) locus. AB - The Wilms tumor suppressor gene WT1 plays a crucial role in the etiology of various human diseases as well as in the development of specific organs including the kidneys, gonads and the spleen. At present the human as well as the Fugu wt1 locus have been characterized. We have used a PAC clone to analyze the murine wt1 locus and report here the structure of the wt1 gene as well as a characterization of the nine wt1 introns regarding their size and sequence at the exon/intron and intron/exon boundaries. In addition we provide a restriction map of the murine wt1 locus which should prove useful for the cloning of various constructs designed for the generation of mouse models. Prompted by the existence of a WT1 antisense transcript in humans we also examined strand-specific transcription at the murine wt1 locus. Our analysis suggests that there is no detectable antisense transcription of sequences within or immediately downstream of wt1 exon 1. We find, however, evidence for a divergent transcript which encompasses sequences at and around minor transcriptional initiation sites of wt1 and which is transcribed in the opposite direction. Despite the very high degree of similarity between the human and the murine wt1 sequence and expression as well as the presence of divergent transcripts in both cases, the existence of antisense transcription does not seem to be conserved between the two species. PMID- 11733137 TI - TaCRK3 encodes a novel Theileria annulata protein kinase with motifs characteristic of the family of eukaryotic cyclin dependent kinases: a comparative analysis of its expression with TaCRK2 during the parasite life cycle. AB - The TaCRK3 gene from the bovine apicomplexan parasite Theileria annulata, encodes a 46 kDa polypeptide with strong homology to the eukaryotic family of cyclin dependent kinases. TaCRK3 does not show significant alignment with any particular CDK group, other than the Pfmrk kinases from the related apicomplexans Plasmodium falciparum and Plasmodium yoelii. It has a putative bipartite nuclear localization signal and is located to parasite nuclei by IFAT. Protein levels are constitutive throughout differentiation of the intra-lymphocytic macroschizont. This contrasts with the expression pattern of TaCRK2 (Kinnaird et al., 1996, Mol. Microbiol., 22, 293-302) which is closely related to the eukaryotic CDK1 /2 families involved in regulation of cell cycle progression. TaCRK2 is also located to the parasite nuclei but has no nuclear localization signal and exhibits transient up-regulation in protein levels during mid-merogony. However compared to TaCRK3, it shows down-regulation near the end of merogony. We predict that TaCRK3 may have a role in regulation of gene transcription while TaCRK2 is more likely to be involved in control of parasite nuclear division. PMID- 11733138 TI - kin-18, a C. elegans protein kinase involved in feeding. AB - TAO1 and TAO2 are recently described protein kinases whose initial characterization has placed them at the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase kinase (MEKK) level of stress-responsive MAPK pathways. Because their physiological roles have not been identified, we sought to study their C. elegans homolog to learn more about their functions. kin-18 encodes a previously uncharacterized protein in C. elegans whose catalytic domain shares over 60% identity with TAO1 and TAO2. We demonstrate that KIN-18 is a protein of 120 kDa whose promoter is active in the pharynx and intestine of C. elegans. To learn more about TAO/KIN-18 function, we studied how expression of constitutively active forms of TAO1 or KIN-18 would affect the physiology of intact worms. Strains of C. elegans expressing active forms of TAO1 or KIN-18 exhibit altered pharyngeal electrophysiology as measured by electropharyngeogram. These worms grow more slowly and lay fewer eggs, phenotypes that could result from reduced feeding. We have also identified a C. elegans gene that encodes a protein kinase similar to mammalian MAPK/ERK Kinase (MEK) 4 whose promoter is active in the pharynx. It is phosphorylated by TAO1 in vitro and physically interacts with TAO1. PMID- 11733139 TI - Expression of a novel member of the ATP1G1/PLM/MAT8 family, phospholemman-like protein (PLP) gene, in the developmental process of mouse cerebellum. AB - We have identified a new member of the ATP1G1/PLM/MAT8 family, named phospholemman-like protein (PLP), from a mouse cerebellum cDNA library. The family consists of small transmembrane proteins that modulate the activities of some ion channels. The deduced amino acid sequence of PLP consists of 93 residues that contain the ATP1G1/PLM/MAT8 motif and a single transmembrane domain, and is most similar to the sequence of mouse phospholemman. In situ hybridization analysis showed that the PLP gene is highly expressed in cerebellar granule cells. PLP expression is elevated in the postnatal developing cerebellum. Thus, it may be implicated in the proliferation, differentiation, and axon elongation of granule cells as they mature and migrate to the internal granule layer. PMID- 11733140 TI - Characterization of the amphioxus presenilin gene in a high gene-density genomic region illustrates duplication during the vertebrate lineage. AB - Molecular studies on the pathology of Alzheimer's disease (AD) have revealed that mutations in presenilin genes (PS) account for some familial cases. Although the contribution of these genes to the etiology is clear, their biological function remains obscure. Approaches using model organisms have been hampered by the fact that rodents contain two PS copies in the genome and do not develop the hallmark features associated with AD upon aging. To understand PS function and evolution, we have searched for PS homologous sequences in the genome of a lower chordate, Branchiostoma floridae. We report the structure of a single copy Branchiostoma floridae PS gene, named BfPS, and describe new features at the molecular level. Moreover, molecular phylogenetic analysis suggests that BfPS is orthologous to the vertebrate PS-1 and PS-2 forms. Finally, the analysis of more than 16 kb of genomic DNA encompassing BfPS identified three novel genes, which cluster with BfPS in a high gene-density region. PMID- 11733141 TI - A repressor-response regulator gene pair controlling jadomycin B production in Streptomyces venezuelae ISP5230. AB - A second regulatory gene (jadR(1)) is located immediately upstream of the putative repressor gene (jadR(2)) in the jad cluster for biosynthesis of the antibiotic jadomycin B in Streptomyces venezuelae ISP5230. It encodes a 234-amino acid polypeptide with a sequence resembling those of response regulator proteins in two-component control systems. Features in the conserved C-terminal domain of JadR(1) place the protein in the OmpR-PhoB subfamily of response regulators. In mutants where jadR(1) was deleted or disrupted, jadomycin B was not produced, implying that the gene has an essential role in biosynthesis of the antibiotic. Cloning jadR(1) from S. venezuelae in pJV73A, and introducing additional copies of the gene into the wild-type parent by plasmid transformation gave unstable strains with pJV73A integrated into the chromosome. The transformants initially showed increased production of jadomycin B but gave lower titers as excess copies of jadR(1) were lost; mature cultures stabilized with a wild-type level of antibiotic production. The mutant from which jadR(1) had been deleted could not be transformed with pJV73A. Altering the composition of jadR genes in the chromosome by integration of vectors carrying intact and disrupted copies of jadR(1) and jadR(2) provided evidence that the two genes form a regulatory pair different in function from previously reported two-component systems controlling antibiotic biosynthesis in streptomycetes. PMID- 11733142 TI - A novel strategy for regulated expression of a cytotoxic gene. AB - The tetracycline (Tet) transactivator system is a powerful promoter system to control gene expression. However, expression of a cytotoxic gene in this system has been limited due to the lethal effect caused by low levels of basal expression of the toxic gene. In this report, we describe a novel strategy to express a toxic gene using the Tet system. The barstar gene is placed downstream of a minimal promoter and the barnase gene downstream of the tetracycline responsive element minimal promoter. When barnase is expressed at a basal level, its toxicity in human cell culture is offset by the similar basal level expression of barstar. However, when the barnase expression is induced with the transactivator protein, its overproduction leads to cell death. Therefore, this strategy allows cytotoxicity to be effectively regulated by tetracycline. PMID- 11733143 TI - The human tuftelin gene: cloning and characterization. AB - Tuftelin has been suggested to play an important role during the development and mineralization of enamel. We isolated the full-length human tuftelin cDNA using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (5' RACE and 3' RACE) methods. Sequence analysis of the tuftelin cDNA revealed an open reading frame of 1170 bp encoding a 390 amino acid protein with a molecular mass of 44.3 kDa and an isoelectric point of 5.7. The human tuftelin protein shares 89 and 88% amino acid sequence identity with the bovine and mouse tuftelin, respectively. It contains a coiled-coil region, recently reported to be involved with tuftelin self-assembly and with the interaction of tuftelin with TIP39 (a novel tuftelin interacting protein). Detailed DNA analysis of the cloned genomic DNA revealed that the human tuftelin gene contains 13 exons and is larger than 26 kb. Two alternatively spliced tuftelin mRNA transcripts have now been identified in the human tooth bud, one lacking exon 2, and the other lacking exon 2 and exon 3. Primer extension analysis, corroborated by RT PCR and DNA sequencing, revealed multiple transcription initiation sites. The cloned 1.6 kb promoter region contained several GC boxes and several transcription factor binding sites such as those for activator protein 1 and stimulatory protein 1. Our blast search of the human and mouse expressed sequence tag data bases, as well as our RT-PCR and DNA sequencing results, and a previous study using Northern blot analysis revealed that tuftelin cDNA sequences are also expressed in normal and cancerous non-mineralizing soft tissues, suggesting that tuftelin has a universal function. We have now identified and characterized different alternatively spliced mouse tuftelin mRNAs in several non-mineralizing tissues. These results provide an important baseline for future understanding of the biological role of tuftelin. PMID- 11733144 TI - Molecular characterization of NSD1, a human homologue of the mouse Nsd1 gene. AB - NR-binding SET-domain-containing protein (NSD1) is a mouse nuclear protein containing su(var)3-9, enhancer-of-zeste, trithorax (SET), proline-tryptophan tryptophan-proline (PWWP) and plant homeodomain protein (PHD)-finger domains (Huang et al., EMBO J. 17 (1998) 3398). This protein also has two other distinct nuclear receptor (NR)-interaction domains, called NID(-L) and NID(+L), and acts as both a NR corepressor and a coactivator by interacting directly with the ligand-binding domain of several NRs. Thus, NSD1 is a bifunctional, transcriptional, intermediary factor. We isolated the human homologue (NSD1) of the mouse NSD1 gene (Nsd1), mapped it to human chromosome 5q35, and characterized its genomic structure. NSD1 consists of at least 23 exons. Its cDNA is 8552 bp long, has an 8088 bp open reading frame, contains at least six functional domains (SET, PWWP-I, PWWP-II, PHD-I, PHD-II, and PHD-III) and ten putative nuclear localization signals, and encodes 2696 amino acids. NSD1 shows 86% identity with the mouse Nsd1 at the nucleotide level, and 83% at the amino acid level. NSD1 is expressed in the fetal/adult brain, kidney, skeletal muscle, spleen, and the thymus, and faintly in the lung. Two different transcripts (9.0 and 10.0 kb) were consistently observed in various fetal and adult tissues examined. These findings favor the character of NSD1 as a nucleus-localized, basic transcriptional factor and also a bifunctional transcriptional regulator, such as that of the mouse Nsd1. It remains to be investigated whether mutations of NSD1 lead to a specific phenotype in man. PMID- 11733145 TI - Structural organization of lower marine nonvertebrate calmodulin genes. AB - The troponin C (TnC) superfamily genes generally possess five introns, and the positions where they are inserted are well conserved except for the fourth intron. Based on a structural comparison of TnC genes, we proposed that the common ancestor of TnC or TnC superfamily genes had no intron corresponding to the modern fourth intron, and therefore members of the superfamily have gained the fourth intron independently within each lineage. Here, we cloned calmodulin (CaM, one of the members of the TnC superfamily) cDNAs from two lower marine nonvertebrates, the sea anemone, Metridium senile, belonging to the Cnidaria, and the sponge, Halichondria okadai, belonging to the Porifera, and also determined their genomic organization. Chordate CaM genes generally possess five introns, but neither sea anemone nor sponge CaM has anything corresponding to the fourth intron of chordate CaMs, suggesting that the early metazoan CaM must have had only four introns. The modern fourth intron of chordate CaMs was acquired within the chordate lineage after nonvertebrate/chordate divergence. This notion concurs with our proposal explaining the evolution of the TnC superfamily genes. PMID- 11733146 TI - Identification of human FEM1A, the ortholog of a C. elegans sex-differentiation gene. AB - We report the isolation, genomic structure, chromosomal location, and expression pattern of the FEM1A gene, the human ortholog of the Caenorhabditis elegans fem-1 and mouse Fem1a genes. The coding sequence is 1851 bp and encodes a 617 amino acid protein. The human FEM1A protein has 65% identity with the mouse Fem1a protein and 34% identity with the C. elegans fem-1 protein, indicating conservation of this protein. The N-terminal region of the encoded protein contains six ankyrin repeat elements, a motif found in signaling and transcriptional regulatory molecules such as Notch and glp1. The gene was highly expressed in human kidney and cardiac tissue, and was expressed at lower levels in multiple tissues, including cartilage. FEM1A was localized to chromosome 5q23.1, a region of conserved synteny with a portion of mouse chromosome 17 that contains Fem1a. In C. elegans, fem-1 is involved in a pathway necessary for sex determination. The identification of a human homolog of this conserved gene suggests a potential role for this sex-determining molecule in humans. PMID- 11733148 TI - Tissue harmonic imaging sonography of breast lesions: improved margin analysis, conspicuity, and image quality compared to conventional ultrasound. AB - The purpose of this preference study is to determine if tissue harmonic imaging (THI) is preferred over conventional sonography for imaging breast masses. A prospective evaluation of 73 identical image pairs (one obtained with conventional sonography, one with THI sonography) was performed, examining 25 cysts, 36 solid masses, and 12 indeterminate lesions. Each image was evaluated for lesion contrast, margins, and overall image quality using a graduated score. Statistical analysis was performed using a modified t test. For cystic and solid lesions, THI was preferred for lesion conspicuity, margin, and overall quality (P<.001). For indeterminate lesions, THI was significantly preferred for lesion conspicuity and overall quality (P<.05), but the preference for margins was not significant. Overall, THI of breast lesions was significantly preferred for lesion contrast and margin evaluation compared to conventional sonography. This modality deserves further evaluation and may improve detection and evaluation of breast lesions. PMID- 11733147 TI - The RUNX3 gene--sequence, structure and regulated expression. AB - The RUNX3 gene belongs to the runt domain family of transcription factors that act as master regulators of gene expression in major developmental pathways. In mammals the family includes three genes, RUNX1, RUNX2 and RUNX3. Here, we describe a comparative analysis of the human chromosome 1p36.1 encoded RUNX3 and mouse chromosome 4 encoded Runx3 genomic regions. The analysis revealed high similarities between the two genes in the overall size and organization and showed that RUNX3/Runx3 is the smallest in the family, but nevertheless exhibits all the structural elements characterizing the RUNX family. It also revealed that RUNX3/Runx3 bears a high content of the ancient mammalian repeat MIR. Together, these data delineate RUNX3/Runx3 as the evolutionary founder of the mammalian RUNX family. Detailed sequence analysis placed the two genes at a GC-rich H3 isochore with a sharp transition of GC content between the gene sequence and the downstream intergenic region. Two large conserved CpG islands were found within both genes, one around exon 2 and the other at the beginning of exon 6. RUNX1, RUNX2 and RUNX3 gene products bind to the same DNA motif, hence their temporal and spatial expression during development should be tightly regulated. Structure/function analysis showed that two promoter regions, designated P1 and P2, regulate RUNX3 expression in a cell type-specific manner. Transfection experiments demonstrated that both promoters were highly active in the GM1500 B cell line, which endogenously expresses RUNX3, but were inactive in the K562 myeloid cell line, which does not express RUNX3. PMID- 11733149 TI - Supraclinoid carotid dissection in a pediatric patient. AB - The authors present the case of a 13-year-old boy with a left temporal lobe infarction that developed during a 400-m run. Magnetic resonance (MR) angiography showed segmental narrowing of the left supraclinoid internal carotid artery (ICA) and a duplicated left middle cerebral artery (MCA). MR angiographic source images revealed a crescent-shaped left carotid lumen, indicative of a supraclinoid carotid dissection. PMID- 11733150 TI - Frontal sinus mucocele: a rare complication of craniofacial fibrous dysplasia. AB - We present plain radiographic, computed tomographic and magnetic resonance imaging (MRI) findings in a 25-year-old female patient with craniofacial fibrous dysplasia (FD). Although FD has a tendency to involve craniofacial bones in a unilateral fashion, the involvement was bilateral and extensive in this case. An additional feature was the presence of a frontal sinus mucocele, presumably due to the involvement of the sinus recess by the dysplastic process. This complication of the craniofacial FD has been reported very infrequently in the literature. PMID- 11733151 TI - Basal cell adenoma in the parapharyngeal space: MR findings. AB - We report MR findings of a case of basal cell adenoma arising in the parapharyngeal space. On MR imaging, the mass was a well-circumscribed cystic and solid tumor with a thin fibrous capsule and intratumoral hemorrhage. It was confined to the parapharyngeal space, separating from the deep lobe of the parotid gland. The solid portion of the tumor was slightly hypointense/hyperintense to the muscle on T1-/T2-weighted images and was relatively well enhanced on Gd-enhanced T1-weighted images. The cystic contents of the mass were hyperintense on both T1- and T2-weighted images and were proven to be a hemorrhage. PMID- 11733152 TI - Internal mammary artery pseudoaneurysms complicating chest wall infection in children: diagnosis and endovascular therapy. AB - Mycotic internal mammary artery (IMA) pseudoaneurysms are sparsely reported in medical literature. We report imaging findings of IMA pseudoaneurysms secondary to chest wall abscesses (staphylococcal and tuberculous) in two children. Both children were successfully treated by endovascular method thus obviating the need for surgery. PMID- 11733153 TI - Focal Castleman disease of the lung: MRI findings. AB - Castleman disease is an uncommon benign disorder mainly affecting mediastinal lymph nodes and rarely visceral organs. The most typical structural finding is hypervascularity which can be well demonstrated both by CT and MRI. We report MR findings of an unusual case of solitary parenchymal lung involvement. PMID- 11733154 TI - Echinococcal cysts of the liver: a retrospective analysis of clinico-radiological findings and different therapeutic modalities. AB - The clinico-radiological findings and management of 61 patients with proven hepatic echinococcal cysts (HEC) examined over the past 5 years were retrospectively analyzed. The sonography and computed tomography (CT) scan findings were studied before and after therapy. The indications, healing, and complications rates for each therapeutic modality were recorded. There is a predominance of HEC in adult females (female to male ratio, 1.77:1). The majority of patients complained of abdominal pain (39/61; 64.4%), and the majority of cysts were solitary (43/61; 70.5%), localized in the right lobe (47/61; 77.0%), and superficial (57/61; 93.4%). Few cysts were complicated by rupture, intraperitoneal (2/61; 3.2%), or intrabiliary (5/61; 8.2%). Medical treatment consisted of antihelmintic chemotherapy alone in eight patients with an adequate response in seven patients (7/8; 87.5%). Thirty-two patients had open or laparoscopic surgery with a cure rate of 50%. The other 50% had major complications requiring a further adjuvant therapy for a complete cure. Nine patients underwent percutaneous catheter ablation combined with adjuvant chemotherapy; healing was observed in eight patients (8/9; 88.8%). HEC are best treated by nonsurgical minimally invasive techniques combined with adjuvant antihelmintic chemotherapy, while surgery should be reserved for complicated HEC by intraperitoneal rupture. PMID- 11733155 TI - Hepatocellular carcinoma after radiofrequency ablation therapy: dynamic CT evaluation of treatment. AB - The aim of this study is to analyze the dynamic CT findings after radiofrequency (RF) ablation for hypervascular hepatocellular carcinoma (HCC). RF ablation was used for 22 tumors in 20 patients. Peripheral enhancement was noted in 89% of regions within 1 month in the arterial phase. In 1-3 months, peripheral enhancement remained at 56% and was reduced to 22% by 3-6 months. It is difficult to determine the therapeutic result within 3 months due to continued peripheral enhancement in the arterial phase. PMID- 11733156 TI - Ultrasonographic analysis of bladder tumors. AB - To analyze the ultrasonographic features of bladder tumors during transabdominal ultrasonography. Ultrasonograms of consecutive patients diagnosed to have bladder tumors from August 1992 to May 1999 were reviewed retrospectively and correlated with the final histological diagnoses. Various features, including the location, size, multiplicity, shape, surface and presence of calcification, of the bladder tumors were analyzed. Of 109 patients reviewed, 104 had transitional cell carcinoma, 3 adenocarcinoma, 1 carcinosarcoma and 1 prostatic carcinoma. A total of 130 tumors (including 13 multifocal tumors) were detected with 30 (27.5%) patients having more than one tumor in the bladder. Tumor size ranged from 0.5 to 9.7 cm (mean 2.7 cm). The tumor involved the trigone (63, 48.5%), lateral wall (32, 24.6%), posterior wall (17, 13.1%), anterior wall (5, 3.8%) or was multifocal (13, 10%). Sixty-seven (51.5%) tumors were polypoidal, 47 (36.2%) were sessile and 16 (12.3%) plaque-like. The surface was regular in 50 (38.5%) tumors and irregular in 80 (61.5%) tumors. Calcified foci were noted in 54 (41.5%) tumors. There was no statistically significant correlation between ultrasonographic features. The most common appearance is a polypoid lesion arising from the trigone, but there was much variation in the ultrasonographic features of bladder tumors. PMID- 11733157 TI - Brucellar spondylodiscitis: MRI diagnosis. AB - Early diagnosis of brucellar spondylodiscitis is often difficult because of the long latent period. Radiographs of the spine, bone scan, and computed tomography (CT) scan provide insufficient data. Among 25 patients with brucellar spondylodiscitis studied by magnetic resonance imaging (MRI), 9 were in the acute stage and 16 were in the chronic stage. MRI is the investigation method of choice in diagnosing brucellar spondylodiscitis. PMID- 11733158 TI - Intraosseous lipoma of the humeral head: MR appearance. AB - Intraosseous lipoma is the rarest benign primary bone tumor. We report a case of juxtaarticular intraosseous lipoma in the humeral head of a 50-year-old man. Roentgenographic, computed tomographic (CT), magnetic resonance (MR), scintigraphic, and histologic findings of this case are presented. PMID- 11733159 TI - Periosteal chondroma presenting as a subcutaneous mass in the thumb. AB - We describe an unusual case of periosteal chondroma of the distal phalanx in the left thumb presenting as a subcutaneous mass in an 81-year-old woman. Magnetic resonance (MR) imaging demonstrated a subcutaneous lesion with hypointensity on T1-weighted images and marked hyperintensity on T2-weighted images, eroding the underlying cortical bone. Computed tomographic (CT) scans showed an erosive change in the dorsal cortex of the distal phalanx. Bone tumors arising from bone surface should be considered in the differential diagnosis of subcutaneous finger masses. PMID- 11733161 TI - Introduction to the symposium--'Learning about the comparative biomechanics of locomotion and feeding'. PMID- 11733162 TI - Mandible movements in ants. AB - Ants use their mandibles for almost any task, including prey-catching, fighting, leaf-cutting, brood care and communication. The key to the versatility of mandible functions is the mandible closer muscle. In ants, this muscle is generally composed of distinct muscle fiber types that differ in morphology and contractile properties. Fast contracting fibers have short sarcomeres (2-3 microm) and attach directly to the closer apodeme, that conveys the muscle power to the mandible joint. Slow but forceful contracting fibers have long sarcomeres (5-6 microm) and attach to the apodeme either directly or via thin thread-like filaments. Volume proportions of the fiber types are species-specific and correlate with feeding habits. Two biomechanical models explain why species that rely on fast mandible strikes, such as predatory ants, have elongated head capsules that accommodate long muscle fibers directly attached to the apodeme at small angles, whereas species that depend on forceful movements, like leaf cutting ants, have broader heads and many filament-attached fibers. Trap-jaw ants feature highly specialized catapult mechanisms. Their mandible closing is known as one of the fastest movements in the animal kingdom. The relatively large number of motor neurons that control the mandible closer reflects the importance of this muscle for the behavior of ants as well as other insects. PMID- 11733163 TI - Comparison of contractile characteristics of muscle from Holstein and double muscled Belgian Blue foetuses. AB - The aim of the present study was to precise the origin of the particular muscle characteristics of double-muscled cattle by comparing muscle properties of Holstein and double-muscled Belgian Blue (BB) foetuses. Ten 100-day-old foetuses of each genotype were studied. The weight and length of foetuses and the length, weight and area of the Semitendinosus (ST) muscle were analysed. Contractile differentiation of the different fibre types was studied by immunohistochemistry using several monoclonal antibodies raised against different myosin heavy chain isoforms (MHC slow, fast, foetal) and by electrophoresis. Proliferation phase of myoblasts from each genotype was analysed in primary culture. On 100 days of foetal life, the foetuses of both genotypes did not show any significant differences in their weight and length. However, BB cattle already present muscle hypertrophy, which seems to originate from a higher myoblast proliferation observed in primary culture. The use of anti-MHC antibodies shows that ST muscle of BB contained a smaller proportion of primary fibres and a higher proportion of secondary fibres which will give principally fast fibres in adult muscle. Electrophoresis analysis confirms a lower proportion of slow MHC in ST of BB. PMID- 11733164 TI - Biomechanics of fast-start swimming in fish. AB - Fast-starts are high acceleration manoeuvres used by fish. Fast-start escape responses can be initiated by a nervous reflex mediated by a Mauthner neurone and result in the simultaneous activation of muscle along one side of the body. Such starts have traditionally been termed 'C' starts where the fish initially bends to a tight C shape and then subsequently gains propulsion away from the stimulus. However, close examination reveals that propulsion occurs even during the initial phase of body bending. In order to generate this forward propulsion the fish must flex with a rearward travelling wave of body bending, and power generated by the muscles in the central region of the trunk must be delivered to the water through the caudal region of the body. Despite simultaneous muscle activation along the length of the fish, a wave of body bending can result from the interaction of the muscle torque acting to bend the fish and the inertia of the body and water acting to resist this bending. The Mauthner neurone causes the muscle along one side of the fish to be activated immediately prior to shortening and so there is no difference in timing between muscle activation and onset of shortening for this initial contraction along the whole length of the body. Although the fish flexes to tighter curvatures towards its caudal region, the white muscle strain is reduced in this region due to the effect of the geometry of the body and muscle. The myotomal muscle generates greater mass-specific power outputs in the central region of the fish, and greater stresses towards the caudal region and this difference in muscle function can be entirely explained by the change in muscle strain. This change in muscle function enables the high muscle powers to be delivered to the water through the caudal region of the fish where it will cause the fast-start acceleration. PMID- 11733165 TI - Critical swimming speed: its ecological relevance. AB - Critical swimming speed (U(crit)) is a standard measurement to assess swimming capabilities of fishes. To conduct this measurement a fish is introduced into a water tunnel in which the current velocity can be controlled by the investigator. At the beginning of the measurement water velocity is low, approximately 1 body length (BL) s(-1), and is then incrementally increased at prescribed intervals. Fishes tend to maintain their position in the water tunnel against the current until fatigue sets in. The time and velocity at which the fish fatigue are used to calculate the critical swimming speed. This procedure is widely used to assess the effects of environmental conditions and pollutants on fish performance. Since the procedure is conducted in conditions that are far from representing most natural environment experienced by fishes, doubts have been raised about its ecological and ecophysiological relevance. Few studies examined correlations between critical swimming speed and traits that seem to be more ecologically relevant. Positive correlations were found between U(crit) and routine activity, metabolic rates and body size of open water, planktivorous fishes, metabolic rates and body size. These data indirectly suggest ecological relevancy of U(crit), but direct measurements relating U(crit) to reproductive success or survival are required to assess such relevancy. PMID- 11733166 TI - Computational hydrodynamics of animal swimming: boundary element method and three dimensional vortex wake structure. AB - The slender body theory, lifting surface theories, and more recently panel methods and Navier-Stokes solvers have been used to study the hydrodynamics of fish swimming. This paper presents progress on swimming hydrodynamics using a boundary integral equation method (or boundary element method) based on potential flow model. The unsteady three-dimensional BEM code 3DynaFS that we developed and used is able to model realistic body geometries, arbitrary movements, and resulting wake evolution. Pressure distribution over the body surface, vorticity in the wake, and the velocity field around the body can be computed. The structure and dynamic behavior of the vortex wakes generated by the swimming body are responsible for the underlying fluid dynamic mechanisms to realize the high efficiency propulsion and high-agility maneuvering. Three-dimensional vortex wake structures are not well known, although two-dimensional structures termed 'reverse Karman Vortex Street' have been observed and studied. In this paper, simulations about a swimming saithe (Pollachius virens) using our BEM code have demonstrated that undulatory swimming reduces three-dimensional effects due to substantially weakened tail tip vortex, resulting in a reverse Karman Vortex Street as the major flow pattern in the three-dimensional wake of an undulating swimming fish. PMID- 11733167 TI - How muscles accommodate movement in different physical environments: aquatic vs. terrestrial locomotion in vertebrates. AB - Representatives of nearly all vertebrate classes are capable of coordinated movement through aquatic and terrestrial environments. Though there are good data from a variety of species on basic patterns of muscle recruitment during locomotion in a single environment, we know much less about how vertebrates use the same musculoskeletal structures to accommodate locomotion in physically distinct environments. To address this issue, we have gathered data from a broad range of vertebrates that move successfully through water and across land, including eels, toads, turtles and rats. Using high-speed video in combination with electromyography and sonomicrometry, we have quantified and compared the activity and strain of individual muscles and the movements they generate during aquatic vs. terrestrial locomotion. In each focal species, transitions in environment consistently elicit alterations in motor output by major locomotor muscles, including changes in the intensity and duration of muscle activity and shifts in the timing of activity with respect to muscle length change. In many cases, these alterations likely change the functional roles played by muscles between aquatic and terrestrial locomotion. Thus, a variety of forms of motor plasticity appear to underlie the ability of many species to move successfully through different physical environments and produce diverse behaviors in nature. PMID- 11733168 TI - The efficiency of aerodynamic force production in Drosophila. AB - Total efficiency of aerodynamic force production in insect flight depends on both the efficiency with which flight muscles turn metabolic energy into muscle mechanical power and the efficiency with which this power is converted into aerodynamic flight force by the flapping wings. Total efficiency has been estimated in tethered flying fruit flies Drosophila by modulating their power expenditures in a virtual reality flight simulator while simultaneously measuring stroke kinematics, locomotor performance and metabolic costs. During flight, muscle efficiency increases with increasing flight force production, whereas aerodynamic efficiency of lift production decreases with increasing forces. As a consequence of these opposite trends, total flight efficiency in Drosophila remains approximately constant within the kinematic working range of the flight motor. Total efficiency is broadly independent of different profile power estimates and typically amounts to 2-3%. The animal achieves maximum total efficiency near hovering flight conditions, when the beating wings produce flight forces that are equal to the body weight of the insect. It remains uncertain whether this small advantage in total efficiency during hovering flight was shaped by evolutionary factors or results from functional constraints on both the production of mechanical power by the indirect flight muscles and the unsteady aerodynamic mechanisms in flapping flight. PMID- 11733169 TI - Biomechanics and kinematics of limb-based locomotion in lizards: review, synthesis and prospectus. AB - The sprawling pattern of locomotion in lizards is kinematically intriguing and is underpinned by a distinctive pattern of appendicular morphology. The statics of the sprawling posture dictate fundamental design principles, and these place constraints on the three-dimensional kinematics of the limbs and body axis as locomotion is effected. The fore and hind limbs accommodate these constraints and dictates in fundamentally similar, but positionally different ways, resulting in different kinematic profiles for these two appendages. Recent kinematic investigations have helped to clarify earlier generalizations about lizard locomotion and have revealed that kinematic patterns are more variable than was previously supposed. Such analyses, and attendant detailed studies of the anatomy of the locomotor system, promise a new synthesis and enhanced understanding of evolutionary patterns of locomotion of lizards and adjustment to various locomotor substrata and modes of progression. PMID- 11733170 TI - Kinematic parameters of terrestrial locomotion in cursorial (ratites), swimming (ducks), and striding birds (quail and guinea fowl). AB - The importance of size, functional features and morphological features in adaptation for walking in birds were studied. The time and space kinematic parameters of locomotion were compared in two running birds, the ratites (rhea, kiwi, Paleognatiforms), in two swimming birds, (ducks) and two striding birds, (quail and Guinea fowl). The results showed that in the two phases, stance and swing, the time and space parameters worked in opposite ways: the duration of the swing was constant, but its length increased with speed. In contrast, the duration of the stance was correlated to speed, while its length was not (except in ducks). In all the birds, a higher speed was achieved by a decrease of the stance duration, and an increase of the swing length. The kinematic parameters were not used in the same way in all species: There is a size effect and large birds increase their speed mainly by increasing the frequency of their movements and the small species increase mainly their amplitude. Nevertheless, it is not the main factor and morphology, such as swimming adaptation features of the ducks, and behaviour, are important because they modify the mechanical constraints and influence the kinematics parameters. PMID- 11733171 TI - Comparative aspects of gait, scaling and mechanics in mammals. AB - In phylogenetically based systematics, Mammalia is the nomenclatural term which designates the clade stemming from the most recent common ancestry of monotremes and theria [, Sys. Biol. 43 (1994) 497]. Considering that locomotor performance is a prevalent function to provide the necessary conditions to survive and transmit genes, it may be questioned if the diverse types of locomotion exhibited by extant mammals could have played a role in their evolution, or have only followed it. We may look after the structural and behavioural features which are involved in mammal locomotion compared to other tetrapods and test if they fit with the proposed phylogeny. Several factors may be checked: scaling effect in relation to gravitational constraints; geometrical distribution of masses in the body, and relative mechanical role of the limbs in the production of the external forces necessary to forward motion. Classically, it was thought that the fastest gaits used by terrestrial mammals were based upon a unique kind of limb motion co ordination, called asymmetrical gaits, which in turn may be thought to be related to a peculiar neuronal wiring. Kinematic analysis brings an insight to this topic. Is the search for an ancestral mammalian locomotor pattern judicious? Notice the small size of many of the first mammals and their probable locomotor plasticity. (relation between grain size of the elements within the substrate and the organism scale). At a small size, the gravitational constraint is less important, and the distinction between terrestrial and arboreal has probably no sense when the limbs are the principal motor elements. There remains the importance of the geometrical distribution of body elements, the proportions of the limbs and of the head-neck complex, the tail merely as an appendix, a set of factors which may have generated the frame of constraints within which diverse locomotor modes have evolved. PMID- 11733172 TI - Modelisation of an unspecialized quadruped walking mammal. AB - Kinematics and structural analyses were used as basic data to elaborate a dynamic quadruped model that may represent an unspecialized mammal. Hedgehogs were filmed on a treadmill with a cinefluorographic system providing trajectories of skeletal elements during locomotion. Body parameters such as limb segments mass and length, and segments centre of mass were checked from cadavers. These biological parameters were compiled in order to build a virtual quadruped robot. The robot locomotor behaviour was compared with the actual hedgehog to improve the model and to disclose the necessary changes. Apart from use in robotics, the resulting model may be useful to simulate the locomotion of extinct mammals. PMID- 11733173 TI - Modeling three-dimensional sculptures of australopithecines (Australopithecus afarensis) for the Museum of Natural History of Vienna (Austria): the post cranial hypothesis. AB - In March 1999, E. Daynes, a sculptor specializing in fossil hominid reconstruction, asked C. Berge to take over the scientific supervision of the reconstruction of two australopithecine post-crania. The heads had been modeled from two skulls found in Hadar (AL 444-2, AL 417). The sculptures were to be represented in a walking stance. The female proportions (AL 417) are estimated from the skeleton of 'Lucy' (AL 288), and the male proportions (AL 444-2) extrapolated from the female ones. Biomechanical and anatomical data (comparison with great apes and humans) are used to reconstruct both dynamic equilibrium and muscular systems. The reconstruction suggests that the fossils moved the pelvis and shoulders extensively when they walked. The hindlimb muscles (such as adductors, gluteal muscles and calf) are fleshy and not or very little tendinous. As indicated by the Laetoli step prints (belonging to a close and contemporaneous species), the foot is adducted during the walk and the support is internal just before take off. In spite of inevitable approximations, such a reconstruction appears to be particularly helpful to bring out morphological and functional traits of the first hominids which are both close to and different from modern humans. PMID- 11733174 TI - Avian long bones, flight and bipedalism. AB - Morphological, geometrical, chemical and mechanical characteristics of avian long bones are reviewed. Important differences exist between long bones of birds and mammals. Differences are also present in appendicular bones of birds, either between wing bones and leg bones or proximal (stylopodial) long bones and distal (zeugopodial) long bones. Special emphasis is put on pneumatization, in terms of both phylogenetic origin and geometrical and mechanical characteristics linked to it. Cortical thickness, bending strength and flexural Young's modulus were significantly lower in pneumatized bones than in marrow-filled bones. Possible adaptive reasons for the differences shown are discussed. PMID- 11733175 TI - The scaling of locomotor performance in predator-prey encounters: from fish to killer whales. AB - During predator-prey encounters, a high locomotor performance in unsteady manoeuvres (i.e. acceleration, turning) is desirable for both predators and prey. While speed increases with size in fish and other aquatic vertebrates in continuous swimming, the speed achieved within a given time, a relevant parameter in predator-prey encounters, is size independent. In addition, most parameters indicating high performance in unsteady swimming decrease with size. Both theoretical considerations and data on acceleration suggest a decrease with body size. Small turning radii and high turning rates are indices of maneuverability in space and in time, respectively. Maneuverability decreases with body length, as minimum turning radii and maximum turning rates increase and decrease with body length, respectively. In addition, the scaling of linear performance in fish locomotion may be modulated by turning behaviour, which is an essential component of the escape response. In angelfish, for example, the speed of large fish is inversely related to their turning angle, i.e. fish escaping at large turning angles show lower speed than fish escaping at small turning angles. The scaling of unsteady locomotor performance makes it difficult for large aquatic vertebrates to capture elusive prey by using whole-body attacks, since the overall maneuverability and acceleration of small prey is likely to be superior to that of large predators. Feeding strategies in vertebrate predators can be related to the predator-prey length ratios. At prey-predator ratios higher than approximately 10(-2), vertebrate predators are particulate feeders, while at smaller ratios, they tend to be filter feeders. At intermediate ratios, large aquatic predators may use a variety of feeding methods that aid, or do not involve, whole body attacks. Among these are bubble curtains used by humpback whales to trap fish schools, and tail-slapping of fish by delphinids. Tail slapping by killer whales is discussed as an example of these strategies. The speed and acceleration achieved by the flukes of killer whales during tail slaps are higher and comparable, respectively, to those that can be expected in their prey, making tail-slapping an effective predator behaviour. PMID- 11733176 TI - Plasticity and constraints on feeding kinematics in anuran larvae. AB - Tadpoles of the majority of anuran species have tiny, anatomically complex mouths. In most species the larval jaws are keratinized sheaths (beaks) overlying infrarostral cartilages. Surrounding the beak is a flexible oral disc and transverse rows of small, keratinized denticles. We used high-speed videography (250, 500 and 1000 frames per second) of Rana catesbeiana tadpoles to observe the kinematics of these mouthparts in feeding and breathing. Tadpoles can protract and retract their jaws as well as make them wider and narrower with each gape cycle. We demonstrate that during air-breathing, movement of the oral disc helps surfacing tadpoles to capture air quickly by preventing water from coming into the mouth. For our feeding study, we observed tadpoles as they grazed on both clean and algal covered glass surfaces. As the jaws close, the lower beak narrows to a greater degree when it encounters resistance. The denticle rows are used to both anchor the mouth and rasp surfaces during feeding. The hyperkinetic mouth parts of tadpoles permit grazing on non-planar surfaces of variable resistance. A trade-off in having such mobile jaws is loss of stability; no generalized tadpoles can generate great forces with their jaws, which would be necessary to subdue and dismember large tough prey. The feeding system of tadpoles is built out of soft tissues (such as cartilage and keratin) that can be shed (the keratinized sheaths) or remodeled (the underlying infrarostral cartilage) quickly, thus facilitating metamorphosis. PMID- 11733177 TI - The role of cranial kinesis in birds. AB - In birds, the ability to move the upper beak relative to the braincase has been the subject of many functional morphological investigations, but in many instances the adaptive significance of cranial kinesis remains unclear. Alternatively, cranial kinesis may be considered a consequence of the general design of the skull, rather than an adaptive trait as such. The present study reviews some results related to the mechanism and functional significance of cranial kinesis in birds. Quantitative three-dimensional X-ray has shown that in skulls morphologically as divers as paleognaths and neognaths the mechanism for elevation of the upper beak is very similar. One of the mechanisms proposed for avian jaw movement is a mechanical coupling of the upper and the lower jaw movement by the postorbital ligament. Such a mechanical coupling would necessitate upper beak elevation. However, independent control of upper and lower jaw has been shown to occur during beak movements in birds. Moreover, kinematic modeling and force measurements suggests that the maximum extensibility of collagen, in combination with the short distance of the insertion of the postorbital ligament to the quadrato-mandibular articulation do not constitute a block to lower jaw depression. The lower jaw ligaments serve to limit the maximal extension of the mandibula. It is suggested here that cranial kinesis in avian feeding may have evolved as a consequence of an increase in eye size. This increase in size led to a reduction of bony bars in the lateral aspect of the skull enabling the transfer of quadrate movement to the upper jaw. The selective forces favoring the development of a kinetic upper beak in birds may be subtle and act in different ecological contexts. Simultaneous movement of the upper and lower jaw not only increases the velocity of beak movements, but with elevated upper beak also less force is required to open the lower jaw. However, the penalty of increased mobility of elements in a lightweight skull and a large eye is potential instability of skull elements during biting, smaller bite forces and limitations on joint reaction forces. Such a lightly built, kinetic skull may have evolved in animals that feed on small plant material or insects. This type of food does not require the resistance of large external forces on the jaws as in carnivores eating large prey. PMID- 11733178 TI - Jaw muscles and the skull in mammals: the biomechanics of mastication. AB - Among non-mammalian vertebrates, rigid skulls with tight sutural junctions are associated with high levels of cranial loading. The rigid skulls of mammals presumably act to resist the stresses of mastication. The pig, Sus scrofa, is a generalized ungulate with a diet rich in resistant foods. This report synthesizes previous work using strain gages bonded to the bones and sutures of the braincase, zygomatic arch, jaw joint, and mandible with new studies on the maxilla. Strains were recorded during unrestrained mastication and/or in anesthetized pigs during muscle stimulation. Bone strains were 100-1000 micro epsilon, except in the braincase, but sutural strains were higher, regardless of region. Strain regimes were specific to different regions, indicating that theoretical treatment of the skull as a unitary structure is probably incorrect. Muscle contraction, especially the masseter, caused strain patterns by four mechanisms: (1) direct loading of muscle attachment areas; (2) a compressive reaction force at the jaw joint; (3) bite force loading on the snout and mandible; and (4) movement causing new points of contact between mandible and cranium. Some expected patterns of loading were not seen. Most notably, strains did not differ for right and left chewing, perhaps because pigs have bilateral occlusion and masseter activity. PMID- 11733179 TI - Tumor necrosis factor-alpha converting enzyme. AB - Tumor necrosis factor-alpha converting enzyme (TACE/ADAM17/CD156q) is a member of the 'A Disintegrin And Metalloprotease', or ADAM, family. It is a multi-domain, type I transmembrane protein that includes an extracellular zinc-dependent protease domain. TACE expression is largely constitutive, but the surface pool is downregulated following cell activation. Cleavage by TACE generates the soluble forms of tumor necrosis factor, transforming growth factor-alpha, and other proteins from their membrane-bound precursors (a phenomenon termed 'shedding'). The recognition of substrates by TACE is poorly understood, but sites distal to the active site are probably involved, and in at least some cases both enzyme and substrate must be membrane-anchored. Cell-activators increase the rate of shedding. Activator-induced shedding is mediated by intracellular kinase cascades, but how these cascades affect the shedding machinery is unknown. The pharmaceutical industry is attempting to design specific TACE inhibitors to treat inflammatory diseases. PMID- 11733180 TI - p63. AB - The p63 gene, a member of the p53 gene family, is expressed into at least six protein isoforms which are divided into two groups, those containing the transcription activation domain (TA isoforms) and those that do not (Delta N isoforms). The TA isoforms are similar to p53 in that they are able to activate transcription of specific target genes and induce cell cycle arrest and apoptosis. The Delta N isoforms are unable to activate transcription, and act in a dominant negative manner, inhibiting transcription activation by both p53 and TA isoforms. p63 knock-out studies in mice have shown that p63 plays an important role in development rather than in tumour suppression. In humans, mutations in the p63 gene have been linked with several developmental abnormalities. Studies on human tumours suggest an oncogenic function for Delta N isoforms rather than a tumour suppressor function for the TA forms. PMID- 11733181 TI - Low-density lipoprotein receptors in the uptake of tumour photosensitizers by human and rat transformed fibroblasts. AB - Low-density lipoproteins (LDL) increase the selectivity of tumour targeting by drugs, including sensitisers for photodynamic therapy, because of the enhanced expression of specific LDL receptors in many types of transformed as compared with non-transformed cells. This investigation aims at gaining more information on the role of LDL receptors in the accumulation of photosensitizer-LDL complexes by human and rat transformed fibroblasts, and the interference of the photosensitizer with LDL recognition by the specific receptors. Both an amphiphilic hematoporphyrin IX (Hp) and a hydrophobic Zn(II)-phthalocyanine (ZnPc) photosensitizers bind to human LDL with molar ratios of 5-6:1 and 10-12:1, respectively. The hematoporphyrin-LDL complex is accumulated by human HT1080 fibroblasts mainly through the high affinity LDL receptors, while the Zn phthalocyanine-LDL complex is internalised through non specific endocytosis because of changes in the apoB LDL structure induced by phthalocyanine association, as suggested by spectroscopic studies. The uptake of LDL-delivered hematoporphyrin, but not Zn-phthalocyanine, is about 4-fold higher in HT1080 cells stimulated for maximal expression of LDL receptors as compared with non stimulated cells. This difference is abolished by LDL acetylation. Human LDL bound hematoporphyrin and Zn-phthalocyanine are up taken by stimulated and non stimulated 4R rat fibroblasts with similar efficiency. Scatchard plot analysis of human (125)I-LDL binding to 4R cells shows the presence of only low affinity receptors while 350,000 high affinity receptors are expressed per HT1080 cell. It is concluded that a careful evaluation of the lack of conformational changes of LDL is critical for guaranteeing the selectivity and efficiency of photosensitizer delivery to tumour cells. PMID- 11733182 TI - Preeclampsia-associated decrease of potential collagenolytic and gelatinolytic activities in the wall of the umbilical cord vein. AB - Preeclampsia is the most common pathological syndrome associated with pregnancy. It is accompanied by remodelling of the extracellular matrix of the umbilical cord. A decrease of collagen content in the umbilical cord vein was described. This decrease may result from reduced collagen biosynthesis or enhanced collagen degradation. It was decided to evaluate whether or not this phenomenon is associated with alterations in the activities of collagenolytic, gelatinolytic and non-specific proteolytic enzymes that may be involved in collagen degradation, as well as the activity of prolidase which provides proline as a substrate for collagen biosynthesis. Studies were performed on the umbilical cord veins of newborns delivered by healthy mothers and those with preeclampsia. The control vein extract, activated with trypsin, degraded reconstituted collagen fibres (64.4+/-2.9 nmol Hyp x mg(-1) protein), whereas the preeclamptic material demonstrated only a trace activity. The venous wall extract contained a latent form of gelatinase that might have been activated by trypsin and 4 aminophenylmercuric acetate. A decrease in the gelatinolytic and proteolytic activities of preeclamptic vein extract at neutral pH was found. Prolidase activity was almost 3-fold lower in the preeclamptic extract (240.6+/-29.3 nmol Pro x min(-1) x mg(-1) protein) in comparison to the control (608.2+/-63.7 nmol Pro x min(-1) x mg(-1)protein). It was concluded that the umbilical cord vein contains a latent form of gelatinase A. The decrease in prolidase activity may reduce collagen biosynthesis, resulting in a decrease of this protein in the preeclamptic umbilical cord vein. PMID- 11733183 TI - Dimerization of Rhizobium meliloti NifH protein in Saccharomyces cerevisiae cells requires simultaneous expression of NifM protein. AB - Compared to free living diazotrophs, the nitrogenase system of symbiotic microorganisms, like Rhizobium (Synorhizobium) meliloti, was poorly studied. The aim of our research was to investigate whether (by analogy with Klebsiella pneumoniae) the NifM product is required and sufficient to obtain active R. meliloti Fe-protein. We cloned nifH gene of R. meliloti and nifM gene of K. pneumoniae in suitable yeast vectors. When introduced into Saccharomyces cerevisiae cells, both genes were effectively expressed to proteins similar to the native products in its immunoreactivity and apparent molecular mass. The association of R. meliloti NifH protein into dimer structure required co expression of NifM that also conferred stability of NifH polypeptide. However, the NifH protein synthesized in yeast did not show enzyme activity, suggesting that the NifM of K. pneumoniae is incapable of activating the NifH protein of R. meliloti. PMID- 11733184 TI - Identification and characterization of nuclear CD38 in the rat spleen. AB - CD38 is an ectoenzyme, which can produce metabolites with intracellular Ca(2+) mobilizing properties and has multiple immunological functions. However, we have recently shown that CD38 is also localized to the nucleus of rat hepatocyte whereby its metabolite cADPR, is able to mobilize nuclear Ca(2+) stores. In this study, we further characterize the localization of nuclear CD38 in the spleen, an important immune organ. We managed to detect the presence of ADP-ribosyl cyclase activity in the nuclear fraction. With Western blotting, we managed to characterize a 42-45 kDa protein band that is typical of CD38 under reducing and non-reducing conditions. However, as a comparison, other nuclear fractions from tissues like thymus, cardiac muscle and cerebellum yielded an additional 85 kDa protein band under non-reducing conditions. Both protein bands could be blocked with a CD38 blocking peptide. Immunohistochemical studies revealed the expression of CD38 in the marginal zone and in the red pulp. In contrast, the germinal center remained largely immunonegative for CD38. This is the first report of a functionally active ADP-ribosyl cyclase/CD38 in the spleen nuclear fraction. The results here suggest that the presence of CD38 in the nuclear environment might have a corollary to functional and regulatory roles in the nucleus. PMID- 11733185 TI - Rate of endothelial expansion is controlled by cell:cell adhesion. AB - Procedures used to alleviate blood vessel occlusion result in varying degrees of damage to the vascular wall and endothelial denudation. The presence of intact, functioning endothelium is thought to be important in controlling smooth muscle cell growth, and limiting the intimal thickening which results from damage to the vessel wall. Recovery of the endothelium is commonly slow and incomplete, due in part to endothelial lateral cell:cell adhesion, which limits cell migration and proliferation. We have investigated the effect of fibroblast growth factor 2 and vascular/endothelial growth factor on the relationship between the temporal distribution of the junctional adhesion proteins, platelet/endothelial cell adhesion molecule, vascular/endothelial cadherin and plakoglobin, and cellular migration and proliferation in an in vitro model of endothelial expansion. We found that whereas cell:cell junctions were initially disturbed to similar extents by single applications of the growth factors, outward cell migration and proliferation rates were inversely correlated with the speed at which cell:cell junctions were re-established. This occurred very rapidly with vascular/endothelial growth factor treatment and more slowly with fibroblast growth factor-2, resulting in more extensive outward migration and proliferation in response to the latter. Platelet/endothelial cell adhesion molecule and vascular/endothelial cadherin appeared to be associated with cell:cell junctional control of migration and proliferation, while plakoglobin did not contribute. It was concluded that the rate of endothelial expansion in response to growth factors, is limited by the rate of re-association of junctional complexes following initial disruption. PMID- 11733186 TI - Inhibition of camel lens zeta-crystallin by aspirin and aspirin-like analgesics. AB - Camel lens zeta-crystallin was reversibly inhibited to various degrees by aspirin (acetyl salicylic acid) and the aspirin-like analgesics: paracetamol (acetaminophen) and ibuprofen (2-(4-isobutyl phenyl)-propionic acid). Among these, aspirin was the most potent inhibitor, causing nearly complete inhibition in a dose-dependent, but time-independent manner. Analysis of inhibition kinetics revealed that aspirin was uncompetitive inhibitor (K(i) 0.64 mM) with respect to NADPH and non-competitive inhibitor (K(i) 1.6 mM) with respect to the substrate, 9,10-phenanthrenequinone (PQ). Multiple-inhibition analysis showed that aspirin and pyridoxal 5' phosphate (PAL-P), a lysine specific reagent, simultaneously bound to a critical lysine residue located towards the NADPH binding region. Consistent with this, NADPH was able to substantially protect zeta-crystallin against aspirin, whereas PQ did not provide any protection. The results suggested that an essential lysine residue was the locus of aspirin binding. The inhibition of zeta-crystallin by aspirin and aspirin-like analgesics was reversible thus eliminating acetylation as a mechanism for inhibition. Reversible binding of aspirin to this lysine may cause steric hindrance resulting in uncompetitive inhibition with respect to NADPH. PMID- 11733187 TI - Cloning and characterization of a novel human transcription factor AP-2 beta like gene (TFAP2BL1). AB - The AP-2 transcription factor has been shown to play an important role in development, morphogenesis, apoptosis, cell-cycle control and has also been implicated in mammary oncogenesis. Here we report the cloning and characterization of a novel human transcription factor AP-2 like gene (TFAP2BL1), which is located on human chromosome 6p12.1-21.1. The TFAP2BL1 cDNA is 2076 base pairs in length, encoding a 452-amino acid polypeptide related to human Ap 2protein. TFAP2BL1 gene has significantly high homology to transcription factor AP-2 gene of human, mouse, chicken, sheep, fruit fly, and C. elegans at amino acid level. RT-PCR analysis shows its relatively high expression level in adult thymus, prostate, small intestine, skeletal muscle, placenta, brain, and testis tissues. PMID- 11733188 TI - The binding affinity of HMG1 protein to DNA modified by cis-platin and its analogs correlates with their antitumor activity. AB - The antitumor activity of cis-platin is believed to result from its interaction with cellular DNA and subsequent processing of DNA adducts by damage recognition proteins. Among them are the high mobility group (HMG) proteins 1 and 2, which have been hypothesized to mediate the effect of cis-platin. One possibility suggests that the tight binding of HMG1 to DNA adducts blocks the repair of damaged DNA. In order to further evaluate such a mechanism, several cis-platinum complexes with known antitumor activity have been used to treat DNA and the affinity of HMG1 to the DNA adduct induced by each drug was determined. The dissociation constants for the complexes of HMG1 with the platinated probe were obtained by gel mobility shift assays. The antitumor activity of the tested platinum compounds was found to correlate with the binding affinity of HMG1 to the respective drug-DNA adduct. These findings support the view that HMG1 contributes to cytotoxicity of cis-platin by shielding damaged DNA from repair. In addition, they offer a fast test for screening new platinum compounds for antitumor activity. PMID- 11733189 TI - Identification and characterization of AGTRAP, a human homolog of murine Angiotensin II Receptor-Associated Protein (Agtrap). AB - Several classes of cytoplasmic proteins have been found to interact specifically with the carboxyl-terminal cytoplasmic region of the angiotensin II type 1 (AT(1)) receptor to regulate different aspects of AT(1) receptor physiology. The murine Angiotensin II Receptor-Associated Protein (Agtrap) is a new member of them. We have recently cloned a new human gene cDNA that codes for a homolog of the murine Agtrap protein from a human fetal brain cDNA library. The deduced polypeptide product of the cDNA is 22 kDa in size, and its DNA and amino acid sequences are 85 and 77% identical to those of the mouse Agtrap gene, respectively. Hence we have named it the human Angiotensin II Receptor-Associated Protein (AGTRAP) gene. The mRNA of AGTRAP was most abundantly expressed in kidney, heart, pancreas and thyroid. Using the yeast two-hybrid screening of a human fetal brain cDNA library, we have identified a new interaction partner of the human AGTRAP protein, RACK1 (Receptor of Activated Protein C Kinase). The AGTRAP-RACK1 interaction was confirmed by GST fusion protein pull-down assays, co immunoprecipitation and surface plasmon resonance. We suggest that the AGTRAP RACK1 interaction may help to recruit signaling complex to the AT(1) receptor to affect AT(1) receptor signaling. PMID- 11733190 TI - A method for the identification of pseudorabies virus protein and angiotensin AT(1A) receptor mRNA expression in the same CNS neurons. AB - Neurotropic viruses have been used over the last 10 years to map the distribution of chains of synaptically connected neurons in the CNS. The peptide content of infected neurons has been determined in a number of cases immunohistochemically. However, it has been unclear whether specific mRNA can be assessed in virus infected neurons. We have established a technique which enables the identification of viral protein and mRNA in the same neuron. In the present study pseudorabies virus retrogradely transported from the kidney was localised using immunohistochemistry and mRNA for the angiotensin II AT(1A) receptor was detected by hybridisation histochemistry. Virus protein was visualised using an immunohistochemical procedure with diaminobenzidine as the chromogen and the same sections were exposed to radioactively labelled ((35)S) riboprobes, hybridising the angiotensin II AT(1A) receptor. The combination of these two approaches resulted in the identification of neurons shown to project polysynaptically to the kidney and express AT(1A) mRNA. These data provide neuroanatomical support for previous physiological observations that ablation of the lamina terminalis and administration of losartan, the AT(1) receptor antagonist, blocks the inhibition of renal sympathetic nerve activity following centrally injected Ang II in rats and sheep [5]. PMID- 11733191 TI - Investigation of the Ca(2+)-independent form of Ca(2+)/calmodulin-dependent protein kinase II in neurite outgrowth. AB - Neuronal Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) plays important roles in the control of nerve functions in response to intracellular Ca(2+) (for reviews [Annu. Rev. Physiol. 57 (1995) 417-445; Trends Neurosci. 17 (1994) 406-412]). Brief Ca(2+) signals activate CaM kinase II, and stimulate an autophosphorylation of Thr-286 which allows the kinase to maintain its activated state even after the Ca(2+) concentration has returned to basal levels [J. Biol. Chem. 264 (1989) 16759-16763; Neuron 3 (1989) 59-70; J. Biochem. 109 (1991) 137 143]. Autophosphorylation of CaM kinase II occurs in situ, but it occurs relatively quickly, within just a few minutes [Endocrinology 134 (1994) 2245 2250; J. Biol. Chem. 268 (1993) 7863-7867; J. Biol. Chem. 265 (1990) 18055 18058]. In the present study, we investigated the involvement of the autophosphorylated/Ca(2+)-independent form of CaM kinase II in neurite outgrowth. When neuroblastoma Neruo2a (Nb2a) cells expressing the alpha isoform of CaM kinase II (Nb2a/alpha cells) were stimulated by plating, they formed neurites. The autophosphorylation of Thr-286 and appearance of Ca(2+)-independent activity preceded the neurite formation. The effect of mutating of the kinase autophosphorylation site replacing Thr-286 with Ala (alpha T286A kinase) or Asp (alpha T286D kinase) was examined. alpha T286A kinase was not converted to a Ca(2+)-independent form, and alpha T286D kinase had Ca(2+)-independent activity significantly as an autophosphorylated kinase. Cells expressing alpha T286D kinase had much longer neurites than Nb2a/alpha cells, whereas cells with alpha T286A kinase did not form neurites. These results indicated that the Ca(2+) independent form of CaM kinase II autophosphorylated at Thr-286 is involved in neurite outgrowth. PMID- 11733192 TI - Development of a transportable incubator for autoradiographic experiments with positron emitter-labeled tracers in living brain tissues. AB - For autoradiography using positron emitter-labeled tracers in living human brain tissues, samples have to be transported to a positron emission tomography (PET) facility. We have developed a transportable apparatus in which slices are incubated in Krebs-Ringer medium with 95% O(2)/5% CO(2) at 34 degrees C. The incubator comprises a polystyrene foam container, inner chamber, heater, thermostat and battery. The container is sealed and oxygen gas is supplied through a filter (pore size 0.2 microm). The gas is evacuated from the container through the filter, to avoid bacterial contamination. Slices (330 microm) of rat brain were arranged on the nylon net of the inner chamber, and lightly fixed in place by covering them with a fine nylon net stretched over a stainless steel ring. The incubation was carried out at a temperature of 34 degrees C maintained by a heater and thermostat, and the medium was bubbled with 95% O(2)/5% CO(2) from a portable gas cylinder. The temperature of the medium in the container was well controlled (34.0+/-0.5 degrees C) for up to 200 min with no positional differences. In the dynamic autoradiographic experiment, the rate of uptake of [(18)F]2-fluoro-2-deoxy-D-glucose (FDG) in sections preincubated for 45 min in the transportable apparatus was no different from that in the sections kept in the usual incubating apparatus. This apparatus may enable autoradiography using positron emitter-labeled tracers in living human brain tissues. PMID- 11733193 TI - Isolation and characterization of intact mitochondria from neonatal rat brain. AB - Poor outcome after neonatal brain injury may be associated with alterations in mitochondrial function. Thus, isolated mitochondria have been a useful tool in understanding the underlying mechanisms of mitochondrial dysfunction. However, isolation and characterization of mitochondria from neonatal rat brain are not fully described. Thus, the aim of this study was to develop a rapid method for the isolation and characterization of functional mitochondria from neonatal rat brain. Mitochondria were isolated from 7-day-old rat brain weighing approximately 500 mg using a discontinuous Percoll density gradient. Brains were homogenized in 12% Percoll/sucrose buffer and layered onto a 26% Percoll/40% Percoll gradient followed by centrifugation. Four methods were used for assessing mitochondrial integrity and function: (1) electron microscopy to assess the morphology of the mitochondria and to determine the relative purity of the preparation; (2) fluorescence of chloromethyl-X-rosamine (Mito Tracker Red) in mitochondria as an indicator of mitochondrial membrane potential (Delta psi(m)); (3) state 3 and 4 respiration; and (4) protein import into mitochondria using an in vitro synthesized mitochondrial malate dehydrogenase (mMDH). These studies demonstrated that the morphology of mitochondria is maintained with intact outer membranes and well-developed cristae, and Delta psi(m) is preserved. Respiration measurements revealed tightly coupled mitochondria with a respiration control ratio (RCR) of 4.1+/-0.18 (n=6). Import of precursor mMDH into mitochondria increased in a time dependent manner maximizing at 15 min. The results indicate that neonatal brain mitochondria isolated using this method are well coupled, morphologically intact and are capable of protein import across the outer and inner mitochondrial membranes. PMID- 11733194 TI - Rapid PCR-mediated synthesis of competitor molecules for accurate quantification of beta(2) GABA(A) receptor subunit mRNA. AB - We describe a fast and easy method for the synthesis of competitor molecules based on non-specific conditions of PCR. RT-competitive PCR is a sensitive technique that allows quantification of very low quantities of mRNA molecules in small tissue samples. This technique is based on the competition established between the native and standard templates for nucleotides, primers or other factors during PCR. Thus, the most critical parameter is the use of good internal standards to generate a standard curve from which the amount of native sequences can be properly estimated. At the present time different types of internal standards and methods for their synthesis have been described. Normally, most of these methods are time-consuming and require the use of different sets of primers, different rounds of PCR or specific modifications, such as site-directed mutagenesis, that need subsequent analysis of the PCR products. Using our method, we obtained in a single round of PCR and with the same primer pair, competitor molecules that were successfully used in RT-competitive PCR experiments. The principal advantage of this method is high versatility and economy. Theoretically it is possible to synthesize a specific competitor molecule for each primer pair used. Finally, using this method we have been able to quantify the increase in the expression of the beta(2) GABA(A) receptor subunit mRNA that occurs during rat hippocampus development. PMID- 11733195 TI - An in vivo binding assay to determine central alpha(1)-adrenoceptor occupancy using [(3)H]prazosin. AB - An alpha(1) adrenoceptor (alpha(1)-AdR) assay using [(3)H]prazosin binding in mouse brain is described which allows in vivo determination of central alpha(1) AdR occupancy for ligands with alpha(1)-AdR affinity. Binding of [3H]prazosin in rat and mouse brain membranes in vitro was used to characterise the pharmacological profile of alpha(1)-AdRs in order to determine any potential species variations. Saturation and displacement studies yielded comparable affinity and pharmacological profile for [(3)H]prazosin binding in mouse and rat brain homogenates. These studies confirmed the absence of species variation for ligands in central alpha(1)-AdR pharmacology which is in good agreement with previous studies in rat brain. Subsequently, in vivo binding of [(3)H]prazosin in mouse whole brain was used to measure the occupancy of a number of AdR ligands. Timecourse studies revealed that a [3H]prazosin (5 mu Ci/mouse) pretreatment time of at least 20 min following intravenous (i.v.) administration was required for optimal specific binding. Ligands were administered systemically 40 min prior to i.v. administration of radiolabel. The alpha(1)-adrenoceptor ligands prazosin (ED(50)=0.15 mg/kg i.p.), benoxathian (0.52 mg/kg i.p.) and phentolamine (51 mg/kg i.p.) were all able to block in vivo [(3)H]prazosin binding from mouse brain. In addition, receptor occupancy values for a number of compounds including haloperidol (ED(50)=0.83 mg/kg s.c.), clozapine (2.2 mg/kg s.c.) and MDL-100907 [R(+)-alpha-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine methanol], (10 mg/kg s.c.)], which possess high to moderate affinity at alpha(1) adrenoceptors, were also determined. These results suggest that in the mouse, [(3)H]prazosin binding can be used to measure in vivo receptor occupancy of ligands with affinity at central alpha(1)-adrenoceptors. PMID- 11733196 TI - Studying semantics in the brain: the rapid stream stimulation paradigm. AB - Event-related potentials (ERPs) provide information about the temporal course of cognitive processes in the brain. They have proved to be a valuable tool in order to explore semantic aspects of word processing. However, to date, research in this field has been mostly concerned with the study of post-lexical features by means of the N400-paradigm. We introduce here the rapid stream stimulation paradigm, in which stimuli reflecting different levels of linguistic information are presented to subjects at a high rate of stimulation. The present protocol shows in detail how this paradigm can be applied. The application of the rapid stream stimulation paradigm evokes the recognition potential (RP), an ERP component that peaks at around 260 ms after stimuli onset and seems to be reflecting lexical selection processes. Results of studies that revealed the sensibility of the RP to visual-semantic aspects and the location of its neural generators within basal extrastriate areas are reported. Although some research has been conducted with the rapid stream stimulation paradigm much remains still to be done. Some of the possibilities that this paradigm offers are further discussed. PMID- 11733197 TI - Measurement of Ca(2+) currents in intact slow skeletal muscle fibers of the frog by the three-microelectrode technique. AB - The recording of currents passing through calcium channels in intact skeletal muscle fibers presents several difficulties. However, use of the three microelectrode voltage-clamp technique at the end of the fiber provided us with a good approximation of current values in such fibers. Using this technique, we were able to measure the calcium-channel current in slow skeletal muscle fibers of the frog (Rana pipiens) and to quantify the effects of denervation on this current. PMID- 11733198 TI - High stability of mRNAs postmortem and protocols for their assessment by RT-PCR. AB - Measurement of gene expression is a major area of brain research. We report on the remarkable postmortem stability of a selection of brain mRNAs in both fresh and frozen brain tissue. We describe techniques for extracting total RNA, synthesizing cDNAs from the mRNAs, amplifying specific cDNAs by the polymerase chain reaction technique, and quantitating the products. We chose five genes to study: the housekeeping gene cyclophilin; the complement components C3 and C4; the microtubule associated protein-2 (MAP-2); and the strongly inducible cyclooxygenase COX-2. We found little deterioration in total RNA or in any of the mRNAs in postmortem tissue up to 96 h. When tissue was frozen, stored at -70 degrees C for 15 years and then thawed, there was no evidence of deterioration from storage, but there was gradual deterioration post thawing. All the mRNAs were stable for 1-2 h at 4 degrees C following thawing. Cyclophilin, C3 and C4 mRNAs were still stable after 8 h, MAP-2 and COX-2 mRNAs showed significant deterioration between 2 and 4 h, and COX-2 mRNA showed drastic deterioration between 4 and 8 h. The data give no indication of rapid postmortem degeneration of RNA. Reliable mRNA values may be obtained from postmortem brain with long autolysis times provided the tissue has been kept in the cold, and from frozen tissues for 1-2 h after thawing. PMID- 11733199 TI - Analysis of gene expression of aldose reductase and sorbitol dehydrogenase in rat Schwann cells by competitive RT-PCR method using non-homologous DNA standards. AB - Aldose reductase (AR) and sorbitol dehydrogenase (SDH) are the enzymes constituting the polyol pathway, an alternate route of glucose metabolism. A wealth of experimental data has indicated the involvement of the polyol pathway in the pathogenesis of diabetic complications. However, there has been surprisingly little research on the relative abundance of SDH to AR in the tissues affected in diabetes. We therefore developed a competitive RT-PCR system to simultaneously determine the mRNA levels of these two enzymes in small amounts of samples, and studied their expression in Schwann cells isolated from adult rat sciatic nerves. Although both AR and SDH mRNA were expressed in the Schwann cells, the levels of SDH cDNA were much lower than those of AR cDNA. The induction of AR mRNA expression in the Schwann cells under hyperosmotic conditions was similarly detected by Northern blot analysis and our competitive RT-PCR method. The RT-PCR system developed in this study may be a useful tool in ascertaining the relative contributions of AR and SDH to the metabolic derangements resulting from the acceleration of polyol pathway activity in the target organ of diabetic complications. PMID- 11733200 TI - Simultaneous video analysis of the kinematics of opercular movement and electromyographic activity during agonistic display in Siamese fighting fish. AB - Neuroethology seeks to uncover the neural mechanisms underlying natural behaviour. One of the major challenges in this field is the need to correlate directly neural activity and behavioural output. In most cases, recording of neural activity in freely moving animals is extremely difficult. However, electromyographic recording can often be used in lieu of neural recording to gain an understanding of the motor output program underlying a well-defined behaviour. Electromyographic recording is less invasive than most other recording methods, and does not impede the performance of most natural tasks. Using the opercular display of the Siamese fighting fish as a model, we developed a protocol for correlating directly electromyographic activity and kinematics of opercular movement: electromyographic activity was recorded in the audio channel of a video cassette recorder while video taping the display behaviour. By combining computer assisted, quantitative video analysis and spike analysis, the kinematics of opercular movement are linked to the motor output program. Since the muscle that mediates opercular abduction in this fish, the dilator operculi, is a relatively small muscle with several subdivisions, we also describe methods for recording from small muscles and marking the precise recording site with electrolytic corrosion. The protocol described here is applicable to studies of a variety of natural behaviour that can be performed in a relatively confined space. It is also useful for analyzing complex or rapidly changing behaviour in which a precise correlation between kinematics and electromyography is required. PMID- 11733201 TI - Tumor environment: a potent driving force in colorectal cancer? PMID- 11733202 TI - Mind the phenotype gap. PMID- 11733203 TI - Genetic susceptibility for panic and phobic disorders. PMID- 11733204 TI - Cleft lip and palate: new genetic clues. PMID- 11733216 TI - The molecular basis and potential role of survivin in cancer diagnosis and therapy. AB - Despite its genetic complexity and multifactoriality, two processes appear almost universally compromised in cancer: the control of cell proliferation and the regulation of cell lifespan. Survivin is a recently described molecule that has been implicated in both processes, and is overexpressed in most human cancers. The exploitation of the survivin signaling pathway might provide important predictive and prognostic clues in cancer diagnosis, and offer new therapeutic alternatives for cancer treatment. PMID- 11733217 TI - Evidence of oxidative damage in Alzheimer's disease brain: central role for amyloid beta-peptide. AB - Amyloid beta-peptide (Abeta) is heavily deposited in the brains of Alzheimer's disease (AD) patients. Free-radical oxidative stress, particularly of neuronal lipids, proteins and DNA, is extensive in those AD brain areas in which Abeta is abundant. Recent research suggests that these observations might be linked, and it is postulated that Abeta-induced oxidative stress leads to neurodegeneration in AD brain. Consonant with this postulate, Abeta leads to neuronal lipid peroxidation, protein oxidation and DNA oxidation by means that are inhibited by free-radical antioxidants. Here, we summarize current research on phospholipid peroxidation, as well as protein and DNA oxidation, in AD brain, and discuss the potential role of Abeta in this oxidative stress. PMID- 11733218 TI - Chromosomal translocations in sarcomas: prospects for therapy. AB - In the past two decades, cytogenetic and molecular genetic investigations have revealed that sarcomas are frequently characterized by specific chromosomal translocations, often involving genes encoding transcription factors. These translocations result in the expression of chimeric oncoproteins that contain functional domains contributed by each of the parental genes. Functioning as deregulated transcription and signaling factors, these novel proteins contribute to the malignant phenotype of the tumor cell by disrupting the tightly regulated process of target gene expression. Several therapeutic strategies that exploit the tumor-specific nature of these oncogenes are currently being investigated. These targeted approaches seek to manipulate the specific biology of these gene fusions, and along with more traditional therapeutic modalities, could augment current approaches to cancer management. PMID- 11733219 TI - Chromosomal breakage syndromes and the BRCA1 genome surveillance complex. AB - Chromosomal instability can occur when the DNA damage response and repair process fails, resulting in syndromes characterized by growth abnormalities, hematopoietic defects, mutagen sensitivity, and cancer predisposition. Mutations in ATM, NBS1, MRE11, BLM, WRN, and FANCD2 are responsible for ataxia telangiectasia (AT), Nijmegen breakage syndrome, AT-like disorder, Bloom and Werner syndrome, and Fanconi anemia group D2, respectively. This diverse group of disorders is thought to be linked through protein interactions with the breast cancer tumor susceptibility gene product, BRCA1. BRCA1 forms a multi-subunit protein complex referred to as the BRCA1-associated genome surveillance complex (BASC), which includes DNA damage repair proteins such as MSH2-MSH6 and MLH1, as well as ATM, NBS1, MRE11, and BLM. Although still controversial, this finding suggests similarities in the pathogenesis of the human chromosome breakage syndromes and a complementary role for each protein in DNA structure surveillance or damage repair. PMID- 11733220 TI - DNA fusion vaccines against B-cell tumors. AB - DNA vaccination is currently being explored as a potential strategy for combatting cancer. However, tumor antigens are often weak and the immune system of patients may be compromised. For B-cell tumors, immunoglobulin idiotypic antigens provide defined targets but are poorly immunogenic. Fusion of a sequence derived from tetanus toxin to the genes encoding idiotypic determinants has proved highly effective in activating protective anti-tumor immunity. DNA fusion vaccines containing immuno-enhancing sequences can augment and direct immune attack on a range of target antigens. Gene-based fusion vaccines offer ease of manipulation and flexible design to activate effective attack on cancer. PMID- 11733221 TI - The role of the nuclear envelope in Emery-Dreifuss muscular dystrophy. AB - The X-linked form of Emery-Dreifuss muscular dystrophy (X-EDMD) is caused by absence, or greatly reduced amounts, of the inner nuclear-membrane protein, emerin. The autosomal dominant form (AD-EDMD) is caused by missense mutations in lamins A and C, two components of the nuclear lamina that interact directly with emerin. Lamin A/C mutations also cause one form of dilated cardiomyopathy (CMD1A) and one form of limb-girdle muscular dystrophy (LGMD1B), both of which have clinical features in common with EDMD, as well as a rare, unrelated form of lipodystrophy (FPLD). Evidence is now emerging that defective assembly of the nuclear lamina is a feature of all these diseases, although not necessarily the direct cause. Why only heart and skeletal muscle, and possibly connective tissue, are affected in EDMD and why expression of the disease is so extremely variable between individuals remains to be explained. PMID- 11733222 TI - Animal models for respiratory chain disease. AB - Elucidation of the pathogenesis in respiratory chain diseases is of great importance for developing specific treatments. The limitations inherent to the use of patient material make studies of human tissues often difficult and the mouse has therefore emerged as a suitable model organism for studies of respiratory chain diseases. In this review, we present an overview of the field and discuss in depth a few examples of animal models reproducing pathology of human disease with primary and secondary respiratory chain involvement. PMID- 11733223 TI - Visualizing the growth of Alzheimer's A beta amyloid-like fibrils. PMID- 11733225 TI - New approaches to surgery for breast cancer. AB - The surgical management of breast cancer is rapidly evolving towards less invasive procedures. Alternative biopsy techniques, including fine-needle aspiration and core needle biopsy, are replacing excisional biopsy as the treatment standard. Breast conservation therapy is now widely used in place of mastectomy, both for small tumors and for larger tumors that have been downstaged through induction chemotherapy. Less invasive procedures for axillary treatment such as lymphatic mapping and sentinel lymph-node biopsy are being explored in an effort to avoid the morbidity associated with axillary lymph-node dissection. For women who still prefer or need to receive a mastectomy, immediate breast reconstruction with autologous tissue provides an excellent cosmetic outcome that is oncologically sound. This is especially appealing to high-risk women who opt to have a prophylactic mastectomy. High-risk women are also being offered the option of receiving chemopreventive treatment that may reduce their lifetime risk of cancer by almost 50%. These new, less invasive approaches require the close cooperation of a team of physicians,including surgeons, pathologists, radiologists, and medical and radiation oncologists. PMID- 11733226 TI - Pituitary tumors: pathophysiology, clinical manifestations and management. AB - Pituitary tumors are frequently encountered intracranial neoplasms. They present with a variety of clinical manifestations that include symptoms and signs of excessive hormone secretion by the tumor, signs of hormone deficits by the normal pituitary gland and others related to expansion of the tumor mass and the resulting compression of surrounding structures such as the optic chiasm and cranial nerves. Advances in molecular biology, immunocytochemical staining and imaging, and the introduction of new treatment options have improved our understanding of the natural history of these adenomas and their management. Available treatments include surgical, medical and radiation therapy. Although the primary treatment for each tumor type may vary, it is important to consider all available options and select the most applicable for that patient. The interaction of all members of management team, including the primary care provider, the endocrinologist and the neurosurgeon in selecting the treatment course can only improve therapeutic outcome. Regardless of the initial choice of treatment,follow-up of all patients should be maintained indefinitely. The managing physician should be familiar with the natural history and long-term complications of pituitary adenomas, and with the side effects of treatments given over the years. PMID- 11733227 TI - Oncogenic co-operation in beta-cell tumorigenesis. AB - Pancreatic islet neoplasms are rare endocrine tumours. The most common type is of beta-cell origin and is known as insulinoma, which can be either benign or malignant. The majority of insulinomas arise sporadically, but a small proportion develop as part of the hereditary multiple endocrine neoplasia type 1 (MEN1) syndrome. As for many human tumours, the genetic events that occur during the initiation and progression of insulinoma are poorly known. The men1 gene product, menin, is deficient in most hereditary cases, but is not obviously affected in the majority of sporadic tumours. Activation of the proto-oncogenes c-myc and ras has been observed during malignant progression, but their role in tumour initiation remains unproven. To address these questions, transgenic mouse models have been increasingly used to explore molecular and genetic events that might also precipitate human neoplasia. Transgenic mice expressing SV40 large T-antigen (Tag) oncogene in beta-cells develop tumours in a multi-stage progression from hyperplasia, angiogenesis, to solid encapsulated tumours. However, Tag, which inactivates the key tumour suppressors p53 and Rb, is not known to be involved in the pathogenesis of human insulinoma. The proto-oncogene, c-myc is implicated in beta-cell growth in both diabetes and tumorigenesis. Activation of Myc appears to be an early event in progression of human insulinoma. The effect of deregulated Myc expression on adult beta-cells in vivo has recently been investigated by developing transgenic mouse models in which the activity of Myc can be regulated ectopically. Although Myc activation initially promotes both proliferation and apoptosis in pancreatic beta-cells, apoptosis is the predominant outcome, giving rise to islet involution and diabetes. Importantly, inhibiting Myc-induced apoptosis (by co-expression of Bcl-x(L)) leads to significantly enlarged islets, many becoming highly vascularized, hyperplastic and invasive. These results suggest that, in the pancreatic beta-cells, early suppression of apoptosis is essential for the survival of Myc-activated beta-cells and islet neoplasia. PMID- 11733228 TI - An adenoviral vector expressing functional heterogeneous proteins herpes simplex viral thymidine kinase and human interleukin-2 has enhanced in vivo antitumor activity against medullary thyroid carcinoma. AB - To explore a more efficient multi-gene antitumor treatment, we developed an adenoviral vector expressing both herpes simplex virus thymidine kinase (HSVtk) and human interleukin-2 (hIL-2) (AdCMVTKhIL2). Production of hIL-2 is expected to augment antitumor T cell and natural killer cell activity. Two separate cassettes expressing HSVtk and hIL-2, each under the control of the human cytomegalovirus (CMV) immediate early gene promoter, were inserted into the early 1 region of adenovirus type 5. This vector showed similar direct cytotoxicity towards infected rat medullary thyroid carcinoma (rMTC) cells as did the single gene vector, AdCMVtk. rMTC cells infected with the virus in vitro showed high sensitivity to ganciclovir. After infection with AdCMVTKhIL2 at 100 m.o.i. for 1 h, more than 20 000 U hIL-2 were produced during 24 h by 1 × 10(6) rMTC cells on day 2 and day 3. hIL-2 was also detected in the supernatants of primary cultures from tumors treated in vivo by the AdCMVTKhIL2 vector. Infected cells lost their tumorigenicity when transplanted subcutaneously into syngeneic rats, whereas all control animals developed tumors. More than 63% of tumors (19 out of 30 treated tumors) were destroyed when AdCMVTKhIL2 was injected intratumorally, compared with 38% when tumors were treated with AdCMVIL2, and 12% when treated with AdCMVtk, indicating an antitumor effect superior to that of each single vector given alone at the same dosage. These results indicate that the AdCMVTKhIL2 vector efficiently produces both HSVtk and hIL-2, and provides an enhanced antitumor activity. PMID- 11733230 TI - Lung adenocarcinoma and malignant uveitis masquerade syndrome. PMID- 11733232 TI - Topical ketorolac after cataract surgery. PMID- 11733233 TI - Nonpenetrating deep sclerectomy for Sturge-Weber syndrome. PMID- 11733234 TI - Corneal effects of latanoprost and timolol. PMID- 11733235 TI - Silicone punctal plug extrusion. PMID- 11733237 TI - Silicone punctal plug extrusion. PMID- 11733239 TI - Transnasal orbital decompression in severe Graves' ophthalmopathy. PMID- 11733243 TI - Surgery for full-thickness macular holes. PMID- 11733245 TI - Visual sensation in cataract surgery. PMID- 11733246 TI - Retina and vitreous pathology after LASIK. PMID- 11733247 TI - Hydroxychloroquine retinopathy. PMID- 11733249 TI - Floppy eyelid syndrome and mental retardation. PMID- 11733251 TI - Scleral expansion surgery for presbyopia. PMID- 11733252 TI - Trachoma control: the end of the beginning? PMID- 11733253 TI - Scleral expansion bands for presbyopia. AB - OBJECTIVE: To evaluate the safety and effectiveness of surgery using scleral expansion bands. DESIGN: Prospective, noncomparative, small case series. PARTICIPANTS: Six subjects were enrolled. Four subjects received implants in one eye and two subjects received implants in both eyes. METHODS: Implantation of the scleral expansion bands (SEB) was performed using Schachar's standard technique. Subjects were observed at six postoperative examination intervals through 1 year. MAIN OUTCOME MEASURES: The key efficacy measures were distance-corrected near visual acuity (at 40 cm and 60 cm) and subjective amplitude of accommodation (push-up, push-down, and minus lens procedures). RESULTS: Distance visual acuity was similar before and after surgery. The near visual acuity and the subjective amplitude of accommodation were temporarily improved in three eyes. However, in the other five eyes, we did not note any improvement of accommodation or near vision after the scleral expansion surgery. CONCLUSIONS: In this study, the outcome of the SEB intervention was characterized by inconsistent and unpredictable results with a low level of patient satisfaction. PMID- 11733254 TI - Randomized clinical trial of Surodex steroid drug delivery system for cataract surgery: anterior versus posterior placement of two Surodex in the eye. AB - OBJECTIVE: To evaluate safety and antiinflammatory efficacy of placing two Surodex (Oculex Pharmaceuticals, Inc., Sunnyvale, CA) in the eye after cataract surgery in comparison with steroid eyedrops and to compare anterior versus posterior chamber placement. DESIGN: Randomized, masked, controlled trial. PARTICIPANTS: One hundred four eyes of 104 Asian patients undergoing extracapsular cataract extraction with intraocular lens implantation were examined. Of these, 33 eyes of 33 patients served as control eyes (group A). INTERVENTION: Two Surodex pellets were inserted in the anterior chamber (AC) of 35 eyes (group B), and two Surodex pellets were inserted in the ciliary sulcus of 36 eyes (group C) at the conclusion of surgery. Control eyes received neither Surodex nor a placebo implant, but were prescribed conventional 0.1% dexamethasone eyedrops four times daily for 4 weeks. MAIN OUTCOME MEASURES: Anterior chamber flare and cells were graded clinically at the slit lamp. Anterior chamber flare was assessed objectively with the Kowa FC500 Laser Flare Meter (Kowa Co. Ltd, Tokyo, Japan). Intraocular pressure and corneal endothelial specular microscopy with morphometric cell analysis were performed for up to 1 year after surgery. RESULTS: Lower flare meter readings occurred in both Surodex groups at all postoperative visits, as compared with the dexamethasone eyedrop group, with statistical significance at days 4 (P = 0.001), 8 (P = 0.001), and 15 (P = 0.02). No difference in flare occurred between AC and ciliary sulcus placement. Clinical slit-lamp assessment of anterior chamber flare and cells showed no difference between Surodex-treated eyes and dexamethasone-treated eyes. Nine of 33 eyes (27.3%) in group A required steroid augmentation, as opposed to 4 of 71 eyes (5.6%) in groups B and C. Inflammatory symptoms were reduced in the Surodex-treated eyes, with statistical significance for ocular discomfort (P = 0.001), photophobia (P = 0.04), and lacrimation (P = 0.01). No complications occurred with Surodex-treated eyes, and no significant difference in endothelial cell loss was noted between Surodex-treated eyes and dexamethasone-treated eyes up to 1 year after surgery. CONCLUSIONS: Intraocular placement of two Surodex is a safe and effective treatment method to reduce intraocular inflammation after cataract surgery and clearly is superior to eyedrops in reducing inflammatory symptoms and aqueous flare as measured with the laser flare meter. No difference in efficacy between AC placement and ciliary sulcus placement of Surodex was detected in this study. PMID- 11733256 TI - Intraoperative management of posterior capsule tears in phacoemulsification and intraocular lens implantation. AB - OBJECTIVE: This paper presents the incidence, causes, and management of posterior capsule (PC) tears and their postoperative outcomes in a large series of eyes that underwent cataract removal and intraocular lens (IOL) implantation, operated on by one surgeon. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Eighteen thousand four hundred seventy consecutive eyes (1992-1999) were reviewed with the assistance of electronic medical records for incidence of PC tears in patients seeking treatment at an outpatient cataract surgery clinic. INTERVENTION: Eyes in this series underwent continuous curvilinear capsulorrhexis (CCC), nucleofractis phacoemulsification, and IOL implantation under topical anesthesia. Different intraoperative surgical strategies such as posterior continuous curvilinear capsulorrhexis (PCCC) were used in the management of PC tears. MAIN OUTCOME MEASURES: Routine preoperative and postoperative visual and refractive outcomes were recorded, including incidence of lens capsule tears, IOL position, and postoperative complications. RESULTS: Posterior capsule tears occurred in 83 of 18,470 eyes, resulting in an overall incidence of 0.45%. Of these 83 eyes with PC tears, 49% received vitrectomy. Fifty-one of 83 PC tears (61.4%) were amenable to be converted to PCCC. Of these 51 procedures, 50 eyes had PC IOL in-the-bag fixation. One eye had PCCC with optic capture. There were no eyes with dropped nuclei or nuclear fragments requiring pars plana vitrectomy. Seventeen eyes (20.5%) had Neuhann anterior capsule rhexis fixation with the haptics placed in the sulcus and IOL securely in the bag. Ten eyes (12.1%) had the IOL placed in the sulcus, and 5 eyes (6.0%) required anterior chamber IOL fixation. None of the 83 PC tears resulted in clinically evident cystoid macular edema, retinal detachment, or endophthalmitis. CONCLUSIONS: With an intact CCC and with conversion of PC tears to PCCC, in-the-bag fixation of IOLs can be achieved in most eyes. PMID- 11733255 TI - Mechanical detachment of the anterior hyaloid membrane from the posterior lens capsule. AB - OBJECTIVE: To investigate the incidence of spontaneous detachment of the anterior hyaloid membrane from the posterior lens capsule and to explore the possibility of surgical separation of the anterior hyaloid membrane. DESIGN: Prospective nonrandomized interventional comparative trial and comparative human tissue study. PARTICIPANTS: Thirty-eight eyes of 32 patients undergoing primary pars plana vitrectomy combined with cataract extraction and intraocular lens implantation and 3 eye bank eyes for histopathologic examination. METHODS: The anterior hyaloid membrane and posterior lens capsule were observed with a fiberoptic endoscope. Surgical separation of the anterior hyaloid membrane was attempted by either the hydrodissection method before cataract extraction (18 eyes) or the direct suction method after cataract extraction (20 eyes). In eye bank eyes, histopathologic examination was performed with (one eye) and without (two eyes) surgical separation of the anterior hyaloid membrane. MAIN OUTCOME MEASURES: Incidence of spontaneous detachment and success rate of surgical detachment of the anterior hyaloid membrane. RESULTS: No eye displayed spontaneous detachment of the anterior hyaloid membrane. Using the hydrodissection method, complete detachment of the anterior hyaloid membrane was achieved in 10 eyes (55.6%), partial detachment was attained in 2 eyes (11.1%), and detachment was not induced at all in 6 eyes (33.3%). With the direct suction method, the anterior hyaloid membrane was completely separated from the posterior lens capsule in 10 eyes (50%), partially detached in 10 eyes (50%), and not detached at all in any eye. The direct suction method had a significantly higher success rate than the hydrodissection method (P < 0.001, chi-square test). In eye bank eyes, the surgical manipulation induced complete separation of the anterior hyaloid membrane from the lens capsule. CONCLUSIONS: Spontaneous detachment of the anterior hyaloid membrane from the posterior lens capsule is very rare. Under endoscopic observation, surgical detachment of the anterior hyaloid membrane was possible in half of the patients. PMID- 11733258 TI - Otago glaucoma surgery outcome study: long-term follow-up of cases of primary glaucoma with additional risk factors drained by Molteno implants. AB - OBJECTIVE: This study was undertaken to provide data on the long-term results of cases of primary open-angle glaucoma with additional risk factors drained by Molteno implants in the province of Otago, New Zealand. DESIGN: Prospective, noncomparative case series PARTICIPANTS: One hundred thirty eyes of 103 patients drained by Molteno implants as the first surgery between 1984 and 1999. INTERVENTION: Insertion of Molteno implant (Ethicon Inc., Johnson & Johnson, Somerville, NJ) using the Vicryl-tie technique. MAIN OUTCOME MEASURES: Intraocular pressure (IOP), visual acuity, visual field. RESULTS: Insertion of a Molteno implant controlled the IOP at 21 mmHg or less with a probability of 1.0 (95% confidence interval [CI], 0.90, 1.0) at up to 7 or more years after surgery. The mean preoperative visual acuity of 20/160 improved to 20/50 after operation and then declined to 20/55 at 5 years and 20/80 after 10 years. The probability of retaining useful vision (visual acuity, >20/400; visual field, >5 degrees radius) was 0.86 (95% CI, 0.77, 0.94) and 0.78 (95% CI, 0.67, 0.89) at 5 and 7 or more years, respectively, after surgery. CONCLUSIONS: The insertion of Molteno implants using the Vicryl-tie technique in this series of 130 cases of primary open-angle glaucoma with additional risk factors controlled the IOP in all cases. PMID- 11733259 TI - Mycobacterium interface keratitis after laser in situ keratomileusis. AB - PURPOSE: To report the clinical course, management, and outcome of infectious interface keratitis caused by mycobacterium species after laser in situ keratomileusis (LASIK). DESIGN: A small noncomparative interventional case series. PARTICIPANTS: Five eyes in four patients who underwent LASIK in different locations around the world and had culture-positive mycobacterium keratitis develop. INTERVENTION: The LASIK flap was lifted or amputated, samples were submitted for Ziehl-Neelsen acid-fast stain and Lowenstein-Jensen's agar cultures for diagnosis; topical treatment with fortified clarithromycin and amikacin was administered until clinical resolution. MAIN OUTCOME MEASURES: Time periods from onset to diagnosis and from diagnosis to clinical resolution, and the final visual acuity. RESULTS: Onset of symptoms of infection occurred after a mean of 20 days (range, 11 days-6 weeks) after LASIK or an enhancement procedure. Definitive diagnosis was obtained after a mean period of 4.5 weeks (range, 12 days-8 weeks) from onset. Keratitis resolved within 8.4 weeks (range, 1-18 weeks) of treatment with fortified clarithromycin and amikacin. Corticosteroids were found to worsen and prolong the course of disease. In four of five eyes the LASIK flap was amputated. CONCLUSIONS: Mycobacterial keratitis is a potentially vision threatening complication after LASIK, characterized by a long latent period, delayed diagnosis, and a protracted course even under intensive specific antibiotic therapy. Inclusion of specific culture media and staining protocols for mycobacteria, along with aggressive treatment on diagnosis, including lifting or amputating the LASIK flap, culturing, topical fortified clarithromycin and amikacin, while avoiding corticosteroids, may significantly improve resolution of the infection and potentially improve the visual outcome. PMID- 11733260 TI - Corneal irregular astigmatism and contrast sensitivity after photorefractive keratectomy. AB - OBJECTIVE: To investigate the relation between contrast sensitivity and corneal irregular astigmatism in eyes after photorefractive keratectomy (PRK). DESIGN: Prospective observational case series. PARTICIPANTS AND CONTROLS: Seventy-nine eyes without corneal haze 6 months or more after PRK and 30 normal control eyes. METHODS: Videokeratography was taken with TMS-1 (Computed Anatomy Inc. New York, NY), and contrast sensitivity for day and night vision was measured using a Multivision Contrast Tester 8000 (Vistech Consultants, Inc., Dayton, OH). MAIN OUTCOME MEASURES: Regular and irregular astigmatism (asymmetry and higher order irregularity components) were computed using Fourier series harmonic analysis of the videokeratography data. Area under the log contrast sensitivity function (AULCSF) was calculated from the contrast sensitivity data taken at five spatial frequencies. RESULTS: After PRK, AULCSFs were significantly smaller for day vision (P = 0.007, unpaired t test) and night vision (P = 0.020) compared with normal controls. AULCSFs for day vision were significantly negatively correlated with asymmetry (Pearson's correlation coefficient, R = -0.23, P = 0.049) and higher order irregularity (R = -0.33, P = 0.004) components but not with the regular astigmatism component (P > 0.3). AULCSF for night vision correlated significantly with asymmetry (R = -0.31, P = 0.013) and higher order irregularity (R = -0.30, P = 0.016) components but not with the regular astigmatism component (P > 0.3). CONCLUSIONS: Corneal irregular astigmatism after PRK significantly influences contrast sensitivity. PMID- 11733261 TI - Abnormal activation in the visual cortex after corneal refractive surgery for myopia: demonstration by functional magnetic resonance imaging. AB - OBJECTIVE: To try to correlate subjective photophobic symptoms with visual pathway modifications (from the retinal image to the visual cortex) after refractive surgery by exploring brain activation on photic stimulation. DESIGN: Noncomparative case series. PARTICIPANTS: Four subjects reporting discomfort produced by luminance (glare, halos, starbursts, or a combination thereof) in one eye after laser in situ keratomileusis (LASIK) were enrolled. The contralateral myopic eye (control) had no visual impairment and had undergone LASIK without complications or had not had previous surgery. METHODS: Functional magnetic resonance imaging was performed during photic stimulation, delivered by an optical fiber, of the affected and unaffected eyes. RESULTS: Functional magnetic resonance imaging provided evidence that most subjective visual symptoms correlated with anatomic flap abnormalities are associated with a higher signal increase in the visual association cortices compared with a nonsymptomatic eye. CONCLUSIONS: Functional magnetic resonance imaging of the visual cortex may help in exploring the mechanisms involved in glare effects after refractive surgery. PMID- 11733262 TI - Natural history of trachomatous scarring in The Gambia: results of a 12-year longitudinal follow-up. AB - PURPOSE: The sight-threatening complications of trachoma are trichiasis and corneal opacity, and these remain the world's most common cause of preventable blindness. The aim of the study was to investigate the rate of progression of trachomatous conjunctival scarring to trichiasis and corneal opacity and to investigate risk factors for such progression. DESIGN: A 12-year, longitudinal study of the incidence and risk factors for the development of trichiasis and corneal opacity in a cohort of patients initially identified with trachomatous scarring in The Gambia. PARTICIPANTS: Six hundred thirty-nine subjects with some degree of trachomatous scarring were identified from the 1986 survey, and attempts were made to trace all these subjects. METHODS: Successfully traced subjects were interviewed, examined, and graded for trachoma. MAIN OUTCOME MEASURES: (1) Twelve-year rates of progression. (2) Risk factors for progression of disease. RESULTS: Three hundred twenty-six of six hundred thirty-nine (51%) subjects were traced and examined, 108 (17%) had died, and 205 (32%) were lost to follow-up. After 12 years, 6.4% (95% confidence interval [CI], 4.0-9.97) of scarred subjects had trichiasis develop, 5.96% (95% CI, 3.67-9.42) had corneal opacity develop, 16.51% (95% CI, 12.71-21.13) had visual impairment/blindness develop, and 2.5% (95% CI, 1.2-5.0) had corneal visual impairment/blindness develop. Mandinka ethnicity was a risk factor for trichiasis (odds ratio [OR], 4.3; 95% CI, 1.3-14.4), and trichiasis at baseline was a risk factor for corneal opacity (OR, 8.4; 95% CI, 1.8-39.2). History of lid surgery for trichiasis was associated with corneal opacity at follow-up (OR, 4.4; 95%CI, 1.4-14.0). Older age was a significant risk factor for development of trichiasis, corneal opacity, and visual loss (OR, 1.07; 95% CI, 1.01-1.12). Bilateral cataract was present in 40% of traced subjects and was associated with the incidence of visual impairment/blindness (OR, 9.4; 95%CI, 4.5-19.6) CONCLUSIONS: This is the first study to demonstrate the link between trichiasis and future corneal opacity, and it provides the rationale for performing lid rotation surgery on patients with trichiasis who do not yet have corneal opacity. The association between corneal opacity at follow-up and previous surgery among trichiasis patients suggests late presentation as a problem. The planning of surgical services will be aided by the incidence figures generated by this study. PMID- 11733263 TI - Ocular Whipple's disease: earlier definitive diagnosis. AB - PURPOSE: Whipple's disease is a rare, chronic, and multiorgan bacterial disease that predominantly involves the gut and its lymphatic drainage in middle-aged Caucasian men but may involve the eye. It is often difficult to diagnose and treat. We report results of one more polymerase chain reaction (PCR) analysis diagnosed ocular Whipple's disease (OWD) case and the care of three new patients with ocular inflammation caused by OWD. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Three patients. METHODS: Diagnostic PCR was performed on a vitreous sample, medical records of three patients with OWD were reviewed, and a literature review (1907 to present) located 74 OWD cases. Analysis of published OWD cases to seek out factors that contribute to earlier, definitive diagnosis and treatment of OWD is made. MAIN OUTCOME MEASURES: Response of ocular inflammation and vision to treatment. RESULTS: One of 3 new and 4 of 74 published OWD cases reviewed were diagnosed with PCR on vitreous samples and subsequently successfully treated with antibiotics without devastating central nervous system (CNS) sequelae. CONCLUSION: Diagnostic PCR on vitreous samples and recognition of key presenting symptoms in patients with OWD allows earlier definitive diagnosis, when the disease may be more amenable to antibiotic treatment, compared with "late" CNS Whipple's disease in the cases reviewed. PMID- 11733264 TI - Chorioretinitis after keratitis caused by Acanthamoeba: case report and review of the literature. AB - OBJECTIVE: A clinicopathologic study to evaluate the histopathologic features associated with Acanthamoeba keratitis and chorioretinitis. DESIGN: Retrospective observational case report. METHODS: On the basis of the clinical history and histologic appearance, the enucleated eye and native corneal button were examined using hematoxylin-eosin stains and special periodic acid-Schiff and Gomori methenamine silver stains. RESULTS: Results of histologic examination of the cornea and retina showed numerous Acanthamoeba cysts in the cornea stromal layers, the necrotic retina, and preretinal and subretinal spaces. CONCLUSIONS: To the authors' knowledge, this is the first proven histologic case of ipsilateral chorioretinitis secondary to primary chronic keratitis caused by Acanthamoeba. The patient had a 30-month history of recurrent keratitis requiring four penetrating keratoplasties. We believe the chorioretinitis resulted from direct spread of the corneal amebic infection. The spread of the Acanthamoeba may have been facilitated by a combined keratoplasty, extracapsular cataract extraction, and intraocular lens insertion. In both specimens, the native corneal button and the enucleated eye with a corneal transplant, the general pathologists overlooked the presence of Acanthamoeba. PMID- 11733265 TI - Circumscribed choroidal hemangioma: clinical manifestations and factors predictive of visual outcome in 200 consecutive cases. AB - PURPOSE: To review the clinical features and management of circumscribed choroidal hemangioma and determine factors predictive of poor visual outcome. DESIGN: Retrospective consecutive noncomparative interventional case series. PARTICIPANTS: Two hundred consecutive patients with circumscribed choroidal hemangioma. MAIN OUTCOME MEASURES: The main outcome measures were analyzed in 155 patients with follow-up of at least 3 months and included complete resolution of subretinal fluid, worsening of visual acuity (more than 2 Snellen lines), and poor final visual acuity (20/200 or worse). RESULTS: The patients were seen at a mean age of 45 years with symptoms of decreased visual acuity (81%), visual field defect (7%), metamorphopsia (3%), floaters (2%), progressive hypermetropia (1%), photopsia (1%), pain (1%), and no symptoms (6%). The referring diagnoses were choroidal hemangioma (29%), choroidal melanoma (29%), choroidal metastasis (9%), retinal detachment (6%), central serous chorioretinopathy (5%), and others. The tumor had a median base of 6.0 mm and median thickness of 3.0 mm. Secondary retinal detachment in the foveal region was present in 81% of the patients. Initial treatment included observation (51%), laser photocoagulation (44%), plaque radiotherapy (4%), external beam radiotherapy (1%), surgical repair of retinal detachment (1%), and enucleation for painful neovascular glaucoma (1%). Kaplan-Meier estimates revealed complete resolution of subretinal fluid in 60% patients at 5 years and 76% patients at 10 years follow-up. By multivariable analysis, clinical factors predictive of complete resolution of subretinal fluid included shorter duration of symptoms (P = 0.03) and inferior quadrant location of tumor (P = 0.001). At initial presentation, 82 of 155 (53%) patients had poor visual acuity (20/200 or worse), and 73 of 155 (47%) patients had good to moderate visual acuity (20/100 or better). Of those 82 patients with poor initial vision, poor final vision was found in 54% at 5 years and 80% at 10 years. Of the 73 patients with good to moderate initial vision, poor final vision was found in 12% at 5 years and 43% at 10 years. By multivariable analysis, clinical factors predictive of poor final visual acuity included poor initial visual acuity (P < 0.001), failure of previous laser photocoagulation before referral (P = 0.01), and tumor management with observation after referral (P = 0.02). Worsening of visual acuity (by more than 2 Snellen lines) was observed in 8% at 5 years and 28% at 10 years of those 82 patients who were initially seen with poor vision. Worsening of visual acuity was found in 10% at 5 years and 30% at 10 years of those 73 patients who initially were seen with good to moderate vision. CONCLUSIONS: Circumscribed choroidal hemangioma is a rare intraocular tumor. In 38% of cases, this tumor is initially misinterpreted before referral as choroidal melanoma or metastasis. Visual acuity is poor in more than 60% of patients at 10 years, despite successful control of associated subretinal fluid in 76% patients. PMID- 11733266 TI - Injection of tissue plasminogen activator into a branch retinal vein in eyes with central retinal vein occlusion. AB - PURPOSE: Central retinal vein occlusion (CRVO) often produces significant and permanent loss of vision in the affected eye. The purpose of this study was to determine if patients with vision loss secondary to CRVO treated with retinal vein cannulation and infusion of tissue plasminogen activator (t-PA) experienced recovery of visual acuity. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Thirty eyes of 30 consecutive patients with CRVO underwent the procedure, but two were subsequently excluded. The remaining 28 eyes of 28 patients with CRVO for an average of 4.9 months before intervention (range, 0.25-30 months) and best-corrected visual acuity 20/63 or worse were included in the study. INTERVENTION: All patients underwent pars plana vitrectomy with cannulation and infusion of t-PA into a branch retinal vein. MAIN OUTCOME MEASURES: Change in visual acuity and the development of complications such as vitreous hemorrhage and neovascular glaucoma were monitored. RESULTS: Twenty-two of 28 patients (79%) experienced at least one line of visual improvement during the follow-up period (average, 11.8 months; range, 3-24 months), and the same number had this level of improvement at the last follow-up examination. Fifteen patients (54%) gained 3 or more lines of acuity within 6 months after the procedure, and 14 (50%) had acuity at last follow-up at least 3 lines better than baseline acuity (average, 6.8 lines). Seven patients had postoperative vitreous hemorrhages ranging from 1 week to 11 months after the procedure; two cleared spontaneously. One patient had a postoperative retinal detachment from a peripheral retinal break that was repaired successfully with pneumatic retinopexy. No other serious intraoperative or early postoperative complications were noted. CONCLUSIONS: Vitrectomy with retinal vein cannulation and infusion of t-PA is a relatively safe procedure that may improve vision in eyes with CRVO. PMID- 11733267 TI - Evaluation of primary surgical procedures for retinal detachment with macular hole in highly myopic eyes: a comparison [corrected] of vitrectomy versus posterior episcleral buckling surgery. AB - PURPOSE: To evaluate postoperative outcomes between pars plana vitrectomy (PPV) and posterior episcleral buckle procedure (PEBP) in myopic eyes with retinal detachment (RD) and macular hole (MH). DESIGN: Retrospective nonrandomized comparative interventional trial. PARTICIPANTS AND INTERVENTION: The study included 30 phakic, highly myopic eyes (from 19-30 negative diopters) of 30 patients with an RD and MH. The patients were divided into two groups: They were assigned to PPV (15 eyes, group A) or to PEBP (15 eyes, group B). No significant (P > 0.01) differences in preoperative visual acuity (VA) between group A and group B eyes were found. Follow-up was 12 months. MAIN OUTCOME MEASURES: Anatomic attachment of the retina was determined, and VA was measured. RESULTS: Retinal reattachment was obtained on 11 of 15 (73.3%) eyes of group A and on 14 of 15 (93.3%) eyes of group B. In group A eyes the VA was substantially unmodified after surgical treatment, whereas in group B eyes the VA observed after surgical treatment increased significantly (P < 0.001) with respect to the preoperative values. CONCLUSIONS: PEBP resulted in better postoperative anatomic and functional results compared with PPV in eyes with extreme degrees of myopia, pronounced posterior staphyloma, and posterior vitreous schisis affected with RD caused by MH. PMID- 11733269 TI - The effect of a growth hormone receptor antagonist drug on proliferative diabetic retinopathy. AB - OBJECTIVE: This study investigated whether the growth hormone receptor antagonist pegvisomant could produce regression of diabetic retinal neovascularization. DESIGN: A prospective, single-group, open-label, phase IIa multicenter trial was conducted. PARTICIPANTS: Twenty-five patients with diabetes mellitus (13 with type 1 and 12 with type 2) and proliferative diabetic retinopathy aged 21 years or older were enrolled. METHODS: Patients received an initial loading dose of 100 mg pegvisomant by subcutaneous injection, followed by self-administered injections of 20 mg daily for 12 weeks. The treatment period was followed by a 12 week period during which patients were observed off treatment. MAIN OUTCOME MEASURE: The effect of treatment on diabetic retinopathy was evaluated from fundus photographs that were graded at a central reading center. RESULTS: Regression of retinopathy was not observed in any patient. At the end of the treatment period, the extent of neovascularization in the study eye was unchanged in 16 patients (9 of 13 with type 1 and 7 of 12 with type 2) and had progressed in 9 patients (4 of 13 with type 1 and 5 of 12 with type 2). The maximum reduction from baseline per patient in the insulin-like growth factor-I (IGF-I) serum level averaged 55%; 17 (68%) of the 25 patients had a greater than 50% reduction in IGF-I. After treatment was discontinued, IGF-I levels returned to baseline. CONCLUSIONS: This study did not find evidence that pegvisomant could produce regression of diabetic retinal neovascularization. PMID- 11733270 TI - Incidence of retinal detachment and visual outcome in eyes presenting with posterior vitreous separation and dense fundus-obscuring vitreous hemorrhage. AB - PURPOSE: To determine visual outcomes and the incidence of retinal detachment in eyes presenting with posterior vitreous separation and dense fundus-obscuring vitreous hemorrhage. DESIGN: Retrospective consecutive noncomparative interventional case series. PARTICIPANTS: Thirty-six eyes (15 right eyes and 21 left eyes) of 34 patients (18 female and 16 male) ranging in age from 42 to 94 years. Mean follow-up was 14 months. METHODS: A comparison of the best-corrected initial visual acuities versus final visual acuities after spontaneous resolution of vitreous hemorrhage or surgical intervention. The number of eyes that were found to have retinal tears or that had a rhegmatogenous retinal detachment develop was documented. Logarithm of the minimum angle of resolution-converted visual acuities was used for comparison. Categorical data were analyzed by Fisher's exact test, and population means were compared by Student's t test. MAIN OUTCOME MEASURES: Final mean visual acuities, number of eyes with at least one retinal tear, location of retinal tears, number of eyes that had retinal detachment develop, and the number of eyes repaired with scleral buckling surgery and/or pars plana vitrectomy. RESULTS: Twenty-four of 36 eyes (67%) were found to have at least one retinal break (range, 0-4 breaks), with 88% of breaks located in the superior retina. Eleven eyes (31%) had more than one retinal break. Fourteen of 36 eyes (39%) had a rhegmatogenous retinal detachment develop that was repaired with pars plana vitrectomy and scleral buckling. An additional 14 eyes (39%) underwent vitrectomy for nonclearing vitreous hemorrhage. The incidence of retinal detachment in eyes with a history of retinal detachment in the contralateral eye was 75% (P = 0.04). Seven of 14 eyes (50%) with retinal detachment had coexisting proliferative vitreoretinopathy. Most retinal breaks and detachments occurred in emmetropic or myopic eyes. For all 36 eyes the mean preoperative visual acuity was 20/1233, and the mean final visual acuity was 20/62 (P < 0.0001). Eyes that had a macula-off retinal detachment develop had worse final visual outcomes (20/264; P = 0.01), as did eyes that had proliferative vitreoretinopathy develop (20/129; P = 0.04). CONCLUSIONS: Acute, spontaneous, nontraumatic posterior vitreous separation with dense fundus obscuring vitreous hemorrhage is associated with a high incidence of retinal tears and detachment. Close follow-up with clinical examination and ultrasonography is necessary, because many of these eyes may eventually require surgical intervention. Aggressive management with early vitrectomy should be considered when there is a history of retinal detachment in the contralateral eye. PMID- 11733271 TI - Dissociated optic nerve fiber layer appearance of the fundus after idiopathic epiretinal membrane removal. AB - PURPOSE: To report the appearance of the fundus, that is seen frequently after removal of an idiopathic epiretinal membrane and which we refer to as the dissociated optic nerve fiber layer appearance. DESIGN: Interventional, noncomparative retrospective case series. PARTICIPANTS: One hundred consecutive patients with an epiretinal membrane who underwent pars plana vitrectomy and epiretinal membrane peeling in one eye. METHODS: Only patients with an idiopathic epiretinal membrane or a membrane associated with a peripheral retinal tear, but without retinal detachment, were considered for this study. Sixty-one patients met these criteria for one eye. Preoperative and postoperative best-corrected visual acuity and preoperative and postoperative blue filter fundus photographs were reviewed. Histopathologic specimens of epiretinal membranes were available for 14 eyes. MAIN OUTCOME MEASURES: The postoperative incidence of the dissociated optic nerve fiber layer appearance on blue filter photographs, visual acuity changes, and the presence of internal limiting membrane in epiretinal membrane specimens. RESULTS: The postoperative incidence of this feature on blue filter fundus photographs was 43%. No difference was found between eyes with or without this feature concerning the average preoperative and postoperative best corrected visual acuity or the average change in visual acuity. Internal limiting membrane was present in all 14 epiretinal membrane specimens available. Five of these 14 patients concerned exhibited a dissociated optic nerve fiber layer appearance and 9 did not. CONCLUSIONS: The dissociated optic nerve fiber layer appearance occurred frequently after removal of an epiretinal membrane. As far as we know, this feature has not been previously reported. It consisted of numerous arcuate striae within the posterior pole in the direction of the optic nerve fibers and slightly darker than the surrounding retina. This feature had no functional effect noticeable by the patient and did not preclude good visual recovery. The small number of histologic samples and the impossibility of quantifying the area of internal limiting membrane peeled off did not allow us to supply proof that this feature is due to the extensive peeling of the internal limiting membrane, although this is the most likely hypothesis. PMID- 11733272 TI - Clinical spectrum of primary ophthalmic rhabdomyosarcoma. AB - PURPOSE: To review the clinical presentation, histopathologic analysis, management, and ocular and systemic outcome of rhabdomyosarcoma affecting the ocular region. DESIGN: Retrospective, noncomparative, consecutive, interventional case series. PARTICIPANTS: Thirty-three consecutive patients with primary ophthalmic involvement of rhabdomyosarcoma from a single tertiary care center specializing in ocular oncology. MAIN OUTCOME MEASURES: Final visual acuity, treatment complications, local recurrence, and distant metastasis. RESULTS: The mean age at presentation was 10 years (median, 7 years; range, 1 month-68 years). At presentation, 8 patients (24%) were older than age 10 years and 4 patients (12%) were older than 20 years. The rhabdomyosarcoma was primarily located in the orbit in 25 cases (76%), conjunctiva in 4 cases (12%), eyelid in 1 case (3%), and uveal tract in 3 cases (9%). Symptoms or signs related to the tumor were present for a mean of 5 weeks and included proptosis in 10 patients (30%), eyelid swelling in 7 patients (21%), and blepharoptosis in 6 patients (18%). The initial diagnosis before referral to us included rhabdomyosarcoma in 8 cases (24%), conjunctivitis in 5 cases (15%), orbital or preseptal cellulitis in 5 cases (15%), idiopathic orbital inflammatory pseudotumor in 4 cases (12%), and others. Using the Intergroup Rhabdomyosarcoma Study Group staging and treatment protocols, the tumor was classified as group I in 4 cases (12%), group II in 12 cases (36%), group III in 16 cases (48%), and group IV in 1 case (3%). Treatment included surgical debulking and various regimens of chemotherapy and radiotherapy for the periocular tumors and enucleation for the three intraocular tumors. Local tumor recurrence was detected in 6 patients (18%). Orbital exenteration was necessary for tumor recurrence in 2 cases (6%). Long-term visual outcome of the 28 patients who maintained their globe was 20/20 to 20/40 in 11 patients (39%), 20/50 to 20/100 in 5 patients (18%), and 20/200 to no light perception in 12 patients (43%). Regional lymph node metastasis was detected in 2 patients (6%), one at initial visit and one after therapy. Distant metastasis occurred in 2 patients (6%), one detected at initial visit and one after therapy. With mean follow-up of 8.3 years, tumor-related death occurred in 1 patient (3%). CONCLUSIONS: Rhabdomyosarcoma can present in the orbit, eyelid, conjunctiva, and uveal tract. After treatment, local tumor recurrence occurs in 18%, metastasis in 6%, and death in 3%. PMID- 11733274 TI - Onset of oscillopsia after visual maturation in patients with congenital nystagmus. AB - PURPOSE: To describe the clinical and oculographic characteristics of a cohort of five patients with congenital nystagmus (CN) and late-onset oscillopsia caused by a coincidental decline in other visual and/or ocular motor functions. DESIGN: Retrospective, observational, case series. PARTICIPANTS: Five visually mature patients with CN and recent-onset oscillopsia were evaluated clinically and with motility recordings. INTERVENTION: Eye movement analysis was performed off-line by computer analysis of digitized data. Nystagmus was analyzed for null-zone characteristics, waveforms, frequency, amplitudes, and slow-phase drift velocity during foveation. Surgical and medical treatment of associated ocular conditions in four of five patients. MAIN OUTCOME MEASURES: Presence of symptomatic oscillopsia and average time during foveation periods of slow-phase drift velocity less than 10 degrees /second. RESULTS: One of the five patients had associated rod-cone dystrophy, and another had recurrence of childhood head posturing with return of an eccentric null zone. The remaining three patients had decompensated strabismus associated with their oscillopsia. All five patients complained of oscillopsia in primary position that was relieved in the four who received treatment. Treatment included prismatic correction in one patient and surgery in three. Recordings in primary position after treatment showed increased duration during foveation periods of slow-phase drift velocity less than 10 degrees /second and an overall decreased intensity (amplitude/frequency) of the nystagmus. CONCLUSIONS: Symptomatic oscillopsia in patients with CN is unusual. This visually disturbing symptom can be precipitated by new or changing associated visual sensory conditions (e.g., decompensating strabismus, retinal degeneration). If the associated conditions can be treated, then accompanying oscillopsia may be relieved. PMID- 11733273 TI - Ocular manifestations of leukemia: leukemic infiltration versus infectious process. AB - OBJECTIVE: To determine whether specific guidelines can be developed to distinguish whether retinal infiltration in leukemia patients represents infection or neoplasia. DESIGN: Retrospective noncomparative interventional case series. PARTICIPANTS: Six patients recently seen at University of California San Francisco with retinal infiltrates in a setting of leukemia, for which adequate written and photographic information of disease course was available. INTERVENTION: Observation consisted of retrospective review of clinic charts, hospital medical records, and fundus photographs. MAIN OUTCOME MEASURES: Determination of whether retinal infiltrates represented neoplasia or infection was made by review of medical records. RESULTS: In this series, neoplastic retinal infiltrates were found in patients who had newly diagnosed leukemia and those who were in blast crisis. In contrast, the two patients who were in complete remission, but had undergone bone marrow transplantation, had retinal infiltrates attributable to infection. CONCLUSIONS: Every patient with retinal infiltrates in the setting of newly or previously diagnosed leukemia requires a systemic and central nervous system workup before the initiation of ophthalmologic treatment. The systemic status of the patient is highly informative in determining whether infection or neoplasia is responsible for the infiltration. PMID- 11733276 TI - Age-based refinement of referral criteria for photoscreening. AB - OBJECTIVE: To evaluate examination results from preschool children referred from photoscreening, and to adjust referral criteria for suspected astigmatism. DESIGN: Cross-sectional study and noncomparative case series. PARTICIPANTS: Thirty-one thousand fifty-three preschool children. METHODS: Analysis of (1) referral rate and unreadable photograph rate for all children screened, (2) examination results and treatment plan for all children referred for suspected astigmatism, and (3) examination results and treatment for all referred children aged less than 1 year. MAIN OUTCOME MEASURES: Referral rate, unreadable photograph rate, predictive value positive, treatment plan. RESULTS: The referral rate dropped from 7.8% for children 6 to 11 months to 5.3% for all other ages. The unreadable photograph rate declined exponentially from 12.1% for children aged 6 to 11 months to 1.1% for children aged 4 years. The predictive value positive of a photoscreen referral for all children in the 6- to 11-month age group was 30%, and only 12 of the 94 referred children were treated. The predictive value positive for children less than 1 year of age referred with suspected astigmatism was even lower (25%), and only one child in this age group was treated. The predictive value positive increased with age, and a higher percentage of older children were treated. For children at least 3 years old referred for suspected astigmatism, the predictive value positive was 67% when the examination was performed by a pediatric ophthalmologist. Strabismus, anisometropia, and high hypermetropia were diagnosed in such patients age 2 and older but never in younger children. CONCLUSIONS: Children less than 1 year of age have a much lower pass rate from photoscreening than do older children because of a higher referral rate and higher unreadable rate in this age group. When these children are examined, significant pathosis is usually absent, and intervention is rarely initiated. Most children age 2 and older who are referred for suspected astigmatism have a high likelihood of significant pathosis. It is probably unnecessary to examine children less than age 2 when their photoscreening suggests only astigmatism; conversely, referrals should still be provided for these children when their screening suggests other potentially amblyogenic factors. PMID- 11733278 TI - Neonatal orbital abscess. AB - PURPOSE: To describe two cases of orbital abscess in neonates and to review the literature of orbital cellulitis in neonates. DESIGN: Two interventional case reports. METHODS: Photographs, orbital computed tomography scans, and full pediatric examination were obtained in two cases of orbital abscess in neonates. RESULTS: Acute ethmoiditis with orbital abscess formation was found in both infants. In one of them, Staphylococcus aureus was identified as the source of infection. CONCLUSIONS: The clinical findings of our cases concur with the literature (eight cases) indicating that orbital abscess caused by Staphylococcal ethmoiditis is the most common form of orbital cellulitis in neonates. PMID- 11733279 TI - Lower eyelid medial canthal tendon laxity grading: an interobserver study of normal subjects. AB - OBJECTIVE: To describe a simple grading system for medial canthal tendon (MCT) laxity and measure its reproducibility. STUDY DESIGN: Observational case series and interobserver variability study. PARTICIPANTS: Fifty subjects (100 lower eyelids) without eyelid disease. The age range was 19 to 98 years. METHODS: Subjects without eyelid pathology or previous surgery were selected. Two ophthalmologists assessed the position of the inferior punctum in relation to the cornea with the patient in primary gaze. Two measurements were made, the first with the lower eyelid at rest and then with the lateral distraction test. Both observers were masked to the other observer's measurements. MAIN OUTCOME MEASURES: Kappa statistics to show the strength of agreement between the two observers for the resting and lateral distraction eyelid positions. RESULTS: High kappa values were found for resting position (agreement in 92 of 100 eyelids) and for lateral distraction testing (agreement in 85 of 100 eyelids). The range of resting positions was from position -1 to position 1. The range of laterally distracted positions was from position 0 to position 5. Overall, a very high level of agreement was achieved between observers. CONCLUSIONS: This grading system for MCT laxity is simple, easy to learn and reproducible, easier than measuring in millimeters, and has an advantage over merely saying the tendon is "lax" or "not lax." We propose this standardized grading system in the evaluation of patients with entropion and ectropion to identify those patients who may benefit from MCT stabilization/plication. PMID- 11733280 TI - Sensitivity of spiral computed tomography scanning for detecting intraocular foreign bodies. AB - OBJECTIVE: The purpose of this study is to determine whether 3-mm computed tomography (CT) cuts are equivalent to 1-mm CT cuts for detecting small intraocular foreign bodies (IOFBs). DESIGN: Experimental instrument validation study. PARTICIPANTS: Seventy-two porcine eyes were divided into three groups. Each group had 0.5 ml of human blood injected into the vitreous cavity and were surgically implanted with IOFBs. INTERVENTION: Each eye was surgically implanted with 0.5-mm metallic, 0.5-mm glass, or 0.5-mm stone IOFBs. Two additional eyes were left without IOFBs to serve as negative controls. MAIN OUTCOME MEASURES: Axial CT images of both 3-mm and 1-mm thickness were obtained and analyzed in masked fashion by two separate neuroradiologists. RESULTS: The sensitivity for detecting 0.5-mm metallic, 0.5-mm glass, and 0.5-mm stone IOFBs with 3-mm CT images was 100%. The sensitivity for detecting 0.5-mm metallic, 0.5-mm glass, and 0.5-mm stone IOFBs with 1-mm CT images was also 100% (confidence interval, 95%; range, 0.88-1.00). Two negative control eyes without IOFBs were also correctly evaluated by the neuroradiologists. CONCLUSIONS: With modern spiral CT scanning, 3-mm cuts are as sensitive as 1-mm cuts for detecting small metallic, glass, and stone IOFBs. PMID- 11733281 TI - New insights into the pathophysiology of primary acquired dacryostenosis. AB - OBJECTIVE: To obtain new insights into the pathophysiology of primary acquired dacryostenosis. DESIGN: Comparative autopsy tissue study with histopathologic correlations. MATERIALS: Tissue specimens from the human nasolacrimal ducts of 36 patients undergoing endonasal dacryocystorhinostomy within a framework of primary acquired dacryostenosis were analyzed by histologic studies and electron microscopic examination. Six lacrimal systems of body donors served as controls. TESTING: One group of tissue specimens from each lacrimal system was prepared and processed with paraffin, sectioned, stained by different methods, and finally examined by light microscopy. The other group was processed with araldite after preparation, sectioned semithin and ultrathin, and examined by transmission electron microscopy. MAIN OUTCOME MEASURES: The degree of dacryostenosis was scored in each tissue specimen by grading the histologic sections as mild (active chronic inflammation), moderate (proliferative sclerotic forms of chronic fibrosis), or severe (total subepithelial fibrosis). RESULTS: Of 36 patients with epiphora, 13 had functional obstruction with a patent lacrimal system on syringing; in 23 cases, the lacrimal passage was completely obstructed. Different pathologic stages correlating to duration of symptoms were found ranging from active chronic inflammation to proliferative sclerotic forms and total subepithelial fibrosis. CONCLUSIONS: Descending inflammation from the eye or ascending inflammation from the nose initiates swelling of the mucous membrane, remodeling of the helical arrangement of connective tissue fibers, malfunctions in the subepithelial cavernous body with reactive hyperemia, and temporary occlusion of the lacrimal passage. In the follow-up, repeated isolated occurrence of dacryocystitis leads to structural epithelial and subepithelial changes, which may lead either to a total fibrous closure of the lumen of the efferent tear duct or to a nonfunctional segment in the lacrimal passage that is manifest on syringing. PMID- 11733282 TI - Endoscopic-guided trephination dacryocystorhinostomy (Hesham DCR): technique and pilot trial. AB - OBJECTIVES: (1) Test the feasibility and the safety of guided transnasal trephination in creating a nasolacrimal fistula. (2) Develop an appropriate lacrimal maintainer and test its value in modulating healing at the fistula site. DESIGN: Prospective, noncomparative interventional case series. PARTICIPANTS: Five cadavers and 19 patients. METHOD: A transcanalicular lacrimal probe penetrated the lacrimal fossa to guide the passage of a flexible trephine up the nose, which created the nasolacrimal communication. A special wide-caliber lacrimal maintainer was inserted along lacrimal tubes within the created passage. OUTCOME MEASURES: For cadaveric study, direct inspection after dissection of the facial flap was performed. For the clinical trial, subjective improvement in watery eye, dye testing, lacrimal probing, lacrimal irrigation, and endoscopic nasal examination. RESULTS: The technique resulted in the creation of a regular fistula of reproducible diameter into which a standard-shaped wide caliber maintainer could be inserted. Three months after removal of the maintainer and 6 months after surgery, a patent ostium was achieved in 17 of 18 (94%) patients who had a completed procedure. Relief of symptoms was achieved in 83%. CONCLUSIONS: Guided endoscopic dacryocystorhinostomy provides a simple and safe option for the treatment of nasolacrimal duct obstruction. The lacrimal maintainer is a useful device to achieve a large patent nasolacrimal communication. PMID- 11733284 TI - Prevalence and causes of visual impairment according to World Health Organization and United States criteria in an aged, urban Scandinavian population: the Copenhagen City Eye Study. AB - PURPOSE: To evaluate the prevalence and causes of visual impairment in an epidemiologic study of aged, urban individuals in Denmark. DESIGN: Cross sectional study. PARTICIPANTS: The study population consisted of 1000 randomly selected residents aged 60 to 80 years in Copenhagen, Denmark. Of 976 eligible persons, 946 (96.9%) could be examined. Information about best-corrected visual acuity (VA) was obtained from 944 cooperative persons (96.7%). METHODS: Data from the Copenhagen City Eye Study were used to assess the cause-specific prevalence of visual impairment as defined by the World Health Organization (WHO) (VA worse than 20/60-20/400 in the better eye) and the criteria used most commonly in the United States (VA worse than 20/40 but better than 20/200 in the better eye). Eligible subjects underwent an extensive ophthalmologic examination at The National University Hospital of Denmark. MAIN OUTCOME MEASURES: Best-corrected VA and primary causes of visual impairment. RESULTS: The prevalence of low vision according to the WHO definition ranged from 2.6% in subjects aged 70 to 74 years to 4.8% in subjects 75 to 80 years of age, with an age-adjusted relative prevalence of 1.58%. Using the U.S. definition, the overall age-adjusted prevalence of visual impairment was 2.9%. The causes of visual impairment according to the WHO criteria were age-related macular degeneration (AMD) (44.4%), cataract (33.3%), glaucoma in combination with cataract (11.1%), myopic macular degeneration (5.6%), and diabetic retinopathy (5.6%). However, according to the U.S. criteria, cataract was the most frequent primary cause (50.0%) and AMD was the second most frequent primary cause (34.4%) of visual impairment. Furthermore, using the U.S. criteria diabetic retinopathy was revealed as equally important as AMD and cataract as a cause of visual impairment among persons aged 65 to 69 years (33.3%). CONCLUSIONS: Increasing age was an independent predictor of visual impairment. Cataract and AMD were the leading causes. Adequate implementation of surgery to treat cataract could reduce visual impairment by 33.3% according to the WHO criteria and by 50% according to the U.S. criteria. PMID- 11733285 TI - Nonlinear behavior of certain optic nerve head parameters and their determinants in normal subjects. AB - OBJECTIVE: To investigate the relationship between the absolute area of certain optic disc parameters and a number of ocular dimensional characteristics and demographic factors in young, healthy subjects. DESIGN: Cross-sectional study. PARTICIPANTS: One hundred twenty-one subjects, aged 16.5 to 35.4 years participated, encompassing a wide range of refractive error from +4 diopters (D) to -25.75 D. One eye from each subject was randomly selected for examination. MAIN OUTCOME MEASURES: Absolute size of the optic disc parameters: optic disc area (DA), neuroretinal rim area (NRA), optic cup area (CA), and peripapillary atrophy area (PA). METHODS: Absolute sizes of optic disc parameters were determined from digitized photographic color transparencies, taking into consideration the magnification of the fundus camera and human eye. Relationships between the absolute size of the optic disc features and axial length, crystalline lens thickness, anterior chamber depth, ocular refraction, front surface keratometry, age, and gender were examined. RESULTS: Axial length, lens thickness, front surface keratometry, and age demonstrated positive associations with all optic disc parameters, and anterior chamber depth and ocular refraction demonstrated negative associations in all cases. Multiple regression analysis revealed that the associations observed univariately for age, anterior chamber depth, lens thickness, front surface keratometry, and ocular refraction were confounded by axial length for all disc parameters. After taking logs, DA and NRA exhibit an exponential relationship with axial length, whereas PA and CA exhibit a linear relationship with axial length. DA and NRA increase by approximately 3% per millimeter increase in axial length at 20 mm and up to 40% for DA and 50% for NRA at an axial length of 35 mm. PA and CA are estimated to increase by 26% (10% 44%) and 10% (5%-16%), respectively, per millimeter increase in axial length. Females have approximately 5% smaller DA and 7% smaller NRA than males, but these effects are not statistically significant (P > 0.2). From the multiple regression analysis, females have 31% larger optic cup areas (1%-57%, P = 0.03) and 24% smaller PA (42% smaller to 5% larger P = 0.06) than males, but these effects are of borderline statistical significance at 5%. CONCLUSIONS: Of the ocular biometric factors considered, axial length seems to be the most important predictor of the absolute area of the optic disc parameters. No association exists between any disc parameter and age in this sample of subjects less than 40 years of age. Females exhibited smaller values for DA, NRA, and PA than did males, differences that were not statistically significant, and larger CA (P = 0.03). PMID- 11733286 TI - Endonasal dacryocystorhinostomy: a report by the American Academy of Ophthalmology. AB - OBJECTIVE: This document describes endonasal dacryocystorhinostomy (DCR) and examines the evidence to answer key questions about the effectiveness of the procedure compared with external DCR; the relative indications, contraindications, advantages, and limitations of the procedure; and patient selection, surgical technique, postoperative care, and complications. METHODS: A literature search conducted for the years 1968 to 2000 retrieved 93 citations. The panel members reviewed 71 of these articles and selected 64 for the panel methodologist to review and rate according to the strength of evidence. A level I rating is assigned to properly conducted, well-designed, randomized clinical trials; a level II rating is assigned to well-designed cohort and case-control studies; and a level III rating is assigned to case series and poorly designed prospective and retrospective studies, including case-control studies. RESULTS: The published literature includes two reports that describe clinical trials comparing endonasal with external DCR with a 1-year follow-up. The success rate was 91% for the external DCR group in both reports and 63% and 75% for the endonasal DCR groups, defined by patency to irrigation. A longitudinal cohort study included a control group of age-matched external DCR patients who were compared with a study group of endonasal DCR patients. The reported success rate at 9 months following surgery for the endonasal cohort was 90% and was not statistically significantly different from the 94% success rate noted in the external DCR control group. Remaining data on reported success rates of primary and revision endonasal DCR were obtained from a collection of uncontrolled observational case studies with varying periods of follow-up and success rates ranging from 59% to 100%. CONCLUSIONS: It is difficult to make definite evidence based determinations about the relative efficacy of endonasal and external DCR because of the deficiencies in the reported literature. Based on level III evidence, the available data suggest that endonasal DCR may be a viable option for the correction of acquired nasolacrimal duct obstruction and complex forms of congenital dacryostenosis in selected patients. This procedure may be indicated on a primary basis or as revisional surgery following failed external or endonasal DCR. Some studies comparing endonasal DCR with external DCR suggested lower success rates in the endonasal group; other studies yielded success rates comparable with or exceeding those of external surgery. Reported complications of endonasal DCR do not generally appear to be greater in frequency or magnitude than those associated with external DCR. Disadvantages of endonasal DCR include the preferred use of general anesthesia by many surgeons, the high cost of expensive equipment and instrumentation, and the relatively steep learning curve for this procedure. Depending on the preference of the surgeon, more postoperative care may be required for patients undergoing endonasal DCR than external DCR. Both the advantages and the limitations of endonasal DCR relative to external DCR should be carefully discussed with patients who are contemplating endonasal surgery. PMID- 11733287 TI - Neurotensin analog NT69L induces rapid and prolonged hypothermia after hypoxic ischemia. AB - OBJECTIVE: To determine whether the neurotensin analog NT69L, administered systemically, could induce mild brain hypothermia after asphyxial cardiac arrest (ACA) in rats. METHODS: The study design was experimental, blinded, randomized, and approved by the animal use committee. All rats had continuous monitoring of brain temperature and sustained 8 minutes of ACA, resuscitation, and either saline or NT69L intravenously after return of spontaneous circulation (ROSC). Rats surviving 14 days after ACA had a neurological deficit score (NDS) and a Morris Water Maze (MWM) test. RESULTS: Seven of eight rats in each group survived 14 days. Brain temperature was less than 35 degrees C 13.1 +/- 3 minutes (mean +/ standard deviation) after NT69L vs controls that remained 37.5 degrees C at the same ambient temperature (p < 0.05 ANOVA). The NT69L group remained below 35 degrees C for 300 +/- 100 minutes while the controls remained at 37.5 +/- 0.5 degrees C. The NDS in the NT69L rats was 3 +/- 3% vs controls 26 +/- 8% (p < 0.05, Kruskal-Wallis, 0% = normal, 100% = brain dead). The NT69L rats performed better on the MWM vs the controls (22 +/- 8 sec vs 45 +/- 26 sec, respectively, p < 0.05 ANOVA). CONCLUSIONS: NT69L induced rapid and prolonged mild brain hypothermia after ACA in this rat model and reduced neurological deficits. PMID- 11733288 TI - Tricyclic antidepressants directly depress human myocardial mechanical function independent of effects on the conduction system. AB - OBJECTIVES: To measure the effect of tricyclic antidepressant drugs (TCAs) on human myocardial contractility. METHODS: Human atrial tissue was obtained during cardiac bypass surgery. The tissue was harvested, suspended in a Tyrode buffer at 37 degrees C, and perfused with a 95%/5% oxygen-carbon dioxide mixture. Developed force was continuously measured using a force transducer and recorded by computer. After an equilibration period, escalating doses of amitriptyline or desipramine were added to the bath. All strips were exposed to the following five concentrations of each drug: 0 (control) 0.4, 4, 40, and 400 microM. The results for each experiment were expressed as the difference between the developed force measured prior to the addition of each concentration of drug and the developed force measured after a 30-minute exposure to the drug. RESULTS: Desipramine decreased the developed force by 27%, 49%, and 74% at concentrations of 0.4, 40, and 400 microM, respectively. Amitriptyline decreased the developed force by 38% at the 40-microM concentration and by 89% at the 400-microM concentration. Untreated strips retained 94% of baseline developed force at 150 minutes. CONCLUSIONS: Tricyclic antidepressants depress human myocardial function in a dose-dependent fashion independent of the effects on the cardiac conduction system. While previous work has demonstrated the effect of therapies for the reversal of impaired cardiac conduction following TCA poisoning, to the best of the authors' knowledge, no reports have documented the effects of therapy on direct myocardial depression. Additional therapies targeted at reversing the direct cardiodepressive effects of TCA may improve outcome following TCA poisoning. PMID- 11733290 TI - Identification of prearrest clinical factors associated with outpatient fatal pulmonary embolism. AB - Massive pulmonary embolism (MPE) is an important cause of outpatient sudden death, and description of these patients is critical for identification and treatment. OBJECTIVE: To test whether MPE patients can be distinguished from patients suffering sudden death from other causes based on clinical, demographic, and historical data. The hypothesis was that MPE cases would be more likely to manifest components of a clinical triad of 1) overt dyspnea, 2) alteration of mental status or syncope, and 3) shock index (pulse divided by systolic blood pressure) >0.8. METHODS: Retrospective case-control study of medical examiner data from 1992 to 1999 including all patients with nontraumatic death, aged 18-65 years, transported to an emergency department, with autopsy-determined cause of death. Analysis was done by 95% confidence interval (95% CI) for difference in proportions and multivariate logistic regression for odds ratios. RESULTS: The MPE patients (n = 37) were younger than the control subjects (n = 347) (40.2 vs 46.5 years, unpaired t-test p < 0.001). At least two of the three components of the triad were present in 56.8% of MPE cases vs 3.5% of controls (95% CI for difference in proportions = 37.3% to 68.0%). Significant variables (and odds ratios) for MPE after multivariate analysis included: dyspnea (13.8), shock index >0.8 (23.6), immobilization or fracture (14.6), seen by a physician within two weeks (5.1), and African American female status (6.4). CONCLUSIONS: Patients in this community with fatal MPE were young with risk factors for pulmonary embolism, and commonly manifested components of a triad including: dyspnea, alteration of mental status/syncope, and shock prior to death. PMID- 11733289 TI - Ethical dilemmas in a randomized trial of asthma treatment: can Bayesian statistical analysis explain the results? AB - OBJECTIVES: The original objective was to determine whether the use of bilevel positive airway pressure (BiPAP) ventilation would reduce the need for endotracheal intubation, the length of hospital stay, and hospital charges in patients with status asthmaticus. The development of physician treatment bias made patient enrollment difficult. The article subsequently describes the use of Bayesian statistics to explain study results when this bias occurs. METHODS: This study was a prospective, randomized controlled clinical trial conducted over a 34.5-month period at an urban university hospital with an emergency department census of 94,000 annual visits. Patients remaining in status asthmaticus after initial standard treatment with inhaled beta-agonists and steroids were randomized to receive BiPAP ventilation plus standard treatment versus standard treatment alone (non-BiPAP), with intubation for either group as needed. Patients with concurrent cardiac or other pulmonary diseases were excluded. The primary outcome measures were endotracheal intubation rate and length of hospital stay. Secondary outcome measures included vital signs (respiratory rate, pulse rate, blood pressure), changes in expiratory peak flow, changes in pulse oximetry values, and hospital charges. Data were analyzed using Fisher's exact test, Mann Whitney tests, and Bayesian statistics. For patients enrolled in the study more than once, data analysis was performed on the first enrollment only. RESULTS: Nineteen patients were enrolled in the BiPAP group and 16 patients in the non BiPAP group. Patients were frequently enrolled more than once and the data from the subsequent enrollments were excluded from the analysis. A marked decrease in enrollment, due to physician treatment bias, led to a premature termination of the study. Demographics showed that the groups were similar in age, sex, initial peak flow rate, and arterial blood gas measurements. There was a 7.3% increase (95% CI = -22 to +45) in the intubation rate in the non-BiPAP group (n = 2) compared with that for the BiPAP group (n = 1). No significant difference was seen in length of hospital stay or hospital charges, although there was a favorable trend toward the BiPAP group. Complications encountered in the BiPAP group included one patient with discomfort associated with the nasal BiPAP mask. Bayesian analysis demonstrated that in order for the collected data to be convincing at the 95% confidence level, the prior conviction among treating physicians that BiPAP was a successful treatment modality would have had to be 98.9%. CONCLUSIONS: In this study, BiPAP appeared to have no deleterious effects in patients with status asthmaticus, with a trend toward decreased endotracheal intubation rate, decreased length of hospital stay, and decreased hospital charges. Although further study with more patients is needed to determine the clinical and statistical significance of this intervention, ethical concerns regarding withholding BiPAP treatment from the patients in the control group forced a premature termination of the study in the authors' institution. PMID- 11733291 TI - Incidence of pericardial effusion in patients presenting to the emergency department with unexplained dyspnea. AB - OBJECTIVE: To evaluate the frequency of pericardial effusion in patients presenting to the emergency department (ED) with unexplained, new onset dyspnea. METHODS: This prospective observational study took place at an urban community hospital ED with a residency program and an annual census of 65,000 visits. Patients presenting between May 1999 and January 2000 with new-onset dyspnea were eligible if they lacked any pulmonary, infectious, hematological, traumatic, psychiatric, cardiovascular, or neuromuscular explanation for their dyspnea after ED evaluation. Patients received a focused echocardiogram by certified emergency physicians. Data were recorded on standardized data sheets and studies were taped for review. Effusions were categorized as small when the fluid stripe measured less than 10 mm. Moderate-sized effusions measured 10 to 15 mm. Large effusions measured more than 15 mm. RESULTS: One hundred three patients were enrolled. Median age was 56 years (IQR 44, 95% CI = 32 to 67). Fourteen patients (13.6%, 95% CI = 8% to 23%) had effusions. Four had large effusions that explained their dyspnea and were admitted to cardiology; two of these effusions were hemorrhagic, and two were viral in origin. Seven patients with small effusions were treated conservatively at home. Three patients had moderate-sized effusions; all were admitted but treated conservatively. CONCLUSIONS: While limited by small numbers, these preliminary data suggest that patients with unexplained dyspnea should be checked for pericardial effusion when bedside ED ultrasound is available. PMID- 11733293 TI - Reliability of the visual analog scale for measurement of acute pain. AB - OBJECTIVE: Reliable and valid measures of pain are needed to advance research initiatives on appropriate and effective use of analgesia in the emergency department (ED). The reliability of visual analog scale (VAS) scores has not been demonstrated in the acute setting where pain fluctuation might be greater than for chronic pain. The objective of the study was to assess the reliability of the VAS for measurement of acute pain. METHODS: This was a prospective convenience sample of adults with acute pain presenting to two EDs. Intraclass correlation coefficients (ICCs) with 95% confidence intervals (95% CIs) and a Bland-Altman analysis were used to assess reliability of paired VAS measurements obtained 1 minute apart every 30 minutes over two hours. RESULTS: The summary ICC for all paired VAS scores was 0.97 [95% CI = 0.96 to 0.98]. The Bland-Altman analysis showed that 50% of the paired measurements were within 2 mm of one another, 90% were within 9 mm, and 95% were within 16 mm. The paired measurements were more reproducible at the extremes of pain intensity than at moderate levels of pain. CONCLUSIONS: Reliability of the VAS for acute pain measurement as assessed by the ICC appears to be high. Ninety percent of the pain ratings were reproducible within 9 mm. These data suggest that the VAS is sufficiently reliable to be used to assess acute pain. PMID- 11733292 TI - Provider adherence to a clinical practice guideline for acute asthma in a pediatric emergency department. AB - Critics of the use of clinical practice guidelines (CPGs) in an emergency department (ED) setting believe that they are too cumbersome and time-consuming, but to the best of the authors' knowledge, potential barriers to CPG adherence in the ED have not been prospectively evaluated. OBJECTIVES: To measure provider adherence to an ED CPG based on National Asthma Education and Prevention Program (NAEPP) recommendations, and to determine factors associated with provider nonadherence. METHODS: Prospective, cohort study of children aged 1-18 years with the diagnosis of an acute exacerbation of asthma who were seen in a pediatric ED and requiring admission, as well as a random selection of children discharged to home following pediatric ED care. The following adherence parameters were assessed: at least three nebulized albuterol treatments in the first hour; early steroid administration (after the first nebulizer treatment); clinical assessments using pulse oximetry and peak expiratory flow (PEF) (for children >6 years old); and use of a clinical score to assess acute illness severity (Asthma Severity Score). Nonadherence was defined as any deviation of the above parameters. RESULTS: Between July 1, 1998, and June 30, 1999, 369 patients were studied. Of these, 38% (139) were discharged to home, 38% (140) were admitted to the observation unit, and 24% (90) were admitted to the inpatient unit. Illness severities at initial presentation to the ED were: 24% (86) had mild exacerbations, 59% (212) had moderate exacerbations, and 17% (62) had severe exacerbations. Sixty-eight percent (95% CI = 63% to 73%) of the patients were managed with complete adherence to the CPG. Of the 32% with some form of nonadherence, most (63%) were children older than 6 years; in this group 64% (48/75) were nonadherent due to lack of PEF assessment. When PEF assessment was disregarded, an 83% (95% CI = 79% to 87%) adherence to the CPG was achieved. Other nonadherence factors included: lack of at least three nebulized albuterol treatments provided timely within the first hour (5%); delay in steroid administration (6%); lack of pulse oximeter use (0.5%); and failure to record clinical score to assess severity (1.1%). Patient age, illness severity (acute and chronic), first episode of wheezing, and high ED volume periods (evenings and weekends) did not worsen adherence. CONCLUSIONS: Clinical practice guidelines can be used successfully in the pediatric ED and provide a more efficient management and treatment approach to acute exacerbations of childhood asthma. With a systematic and concise CPG, barriers to adherence in a pediatric ED appear to be minimal, with the exception of using PEF in the routine ED assessment. PMID- 11733294 TI - Advance directives in skilled nursing facility residents transferred to emergency departments. AB - OBJECTIVE: Ten years have passed since Congress enacted the Patient Self Determination Act to promote the use of advance directives (ADs). This study was performed to determine the frequency, type, demographic distribution, and utility of ADs that accompany residents of skilled nursing facilities (SNFs) transferred to emergency departments (EDs). METHODS: This was an observational, cross sectional cohort of SNF residents, transferred to two urban, academic EDs. Chart review and physician interviews were conducted on consecutive patients arriving during 12-hour data collection shifts. RESULTS: Among 715 patients entered, 315 [44%, 95% confidence interval (95% CI) = 40% to 48%] had an AD. Advance directives were significantly more prevalent among white (50%) than African American (34%) or Hispanic (39%) patients (p < 0.001), and varied from 0% to 94% among SNFs. Of the 315 patients with ADs, do-not-resuscitate (DNR) orders were the most prevalent (65%, 95% CI = 58% to 69%). Although 75% (95% CI = 69% to 81%) of the DNR orders addressed cardiopulmonary resuscitation (CPR), only 12% (95% CI = 8% to 16%) addressed intubation. Among 39 patients who required intubation or CPR, 44% had ADs, 82% (95% CI = 57% to 96%) of which were deemed useful. CONCLUSIONS: Despite a decade of legislation promoting their use, ADs are lacking in most SNF residents transferred to EDs for evaluation and in most settings in which a clinical indication exists for intubation or CPR. Variation in their prevalence appears to be associated with both ethnicity and SNF origin. Although about three-fourths of DNR ADs addressed CPR, only about one in ten offered guidance regarding intubation. When available, ADs are used in most instances to guide emergency care. PMID- 11733295 TI - Risk factors for cardiovascular and cerebrovascular death among African Americans and Hispanics in Los Angeles, California. AB - OBJECTIVE: To describe the risk factors associated with cardiovascular mortality in the African American (AA) and Hispanic populations in Los Angeles County in an effort to define causes for the excess mortality seen in AAs. METHODS: This was a longitudinal analysis of all-cause, cardiovascular, and cerebrovascular mortality in a large, prospective multiethnic cohort of individuals aged 45-74 years. Death rates between AA and Hispanic men and women during the six-year period from 1993 to 1998 due to hypertension, cardiomyopathy, acute myocardial infarction (AMI), ischemic heart disease, and stroke were compared. RESULTS: There were 1,157 deaths due to cardiovascular disease (CVD) or cerebrovascular disease among the 71,798 eligible members of the cohort included in these analyses. Age-adjusted mortality rates were two to five times higher in AAs as compared with Hispanics (e.g., 373.15 in AAs for hypertensive disease vs 50.37 in Hispanics). A history of hypertension was the most common significant risk factor for CVD; other risk factors significantly associated with CVD mortality included cigarette smoking and a past history of diabetes and stroke. Adjusting for these factors did not remove the significance of AA ethnicity as a risk factor for CVD mortality in either subjects reporting or subjects not reporting hypertension at baseline. CONCLUSIONS: The evidence for both higher relative severity and higher incidence of hypertensive disease among AAs, and the consistency of the effect across gender, suggests that a major determinant of risk may be a gene environment interaction. PMID- 11733297 TI - Clinical pearls: palmar rash. PMID- 11733296 TI - Snowmobile injuries in Utah. AB - OBJECTIVE: To describe the epidemiology of snowmobile injuries in Utah. METHODS: Analysis of probabilistically linked statewide emergency department (ED), hospital admission, and death certificate data for 1996 and 1997. RESULTS: There were 625 cases of snowmobile-related injuries. The majority (83%) were evaluated in the ED only. Median ED patient age was 29 years (range 3-74 years), and 66% were male. The leading diagnoses were open wounds to the head (7.8%), back strains (5.4%), and contusions of the trunk and lower extremities (5.2% and 5.0%, respectively). An Injury Severity Score (ISS) of > or = 4 (range 1-75) was found in 37% of the ED patients. The median charge was $373 per patient, with two-year cumulative charges of $266,283. One hundred seven patients required inpatient hospital care. Median inpatient age was 32 years (range 4-92 years), and 60% were male. Leading inpatient diagnoses were fracture of the vertebral column (9.3%), lower extremity fracture (9.3%), upper extremity fracture (6.5%), and pelvis fracture (3.7%). An ISS of > or = 4 (range 1-38) was found in 70% of the hospitalized patients. Average length of stay was 3 days, with a range of 1 to 68 days. Median inpatient charge was $6,003 per patient, with two-year cumulative charges of $1,333,218. Ten inpatients required transfer for rehabilitation or skilled nursing care. There were a total of six fatalities, three of which occurred in the ED, one in the inpatient population, and two identified from the death certificate database. CONCLUSIONS: By combining ED, inpatient, and death certificate data sets, probabilistic linkage provides a comprehensive description of snowmobile-related injuries and a baseline evaluation of morbidity, mortality, and financial burden. PMID- 11733298 TI - Are clinician-investigators Bayesian? PMID- 11733299 TI - Rising to the challenge of family-centered care in emergency medicine. PMID- 11733300 TI - Mental health aspects of emergency medical services for children: summary of a consensus conference. AB - OBJECTIVE: To address the mental health needs of children involved in emergency medical services (EMS). METHODS: A multidisciplinary consensus conference convened to identify mental health needs of children and their families related to pediatric medical emergencies, to examine the impact of psychological aspects of emergencies on recovery and satisfaction with care, and to delineate research questions related to mental health aspects of medical emergencies involving children. RESULTS: The consensus group found that psychological and behavioral factors affect physical as well as emotional recovery after medical emergencies. Children's reactions are critically affected by age and developmental level, characteristics of the emergency medical event, and parent reactions. As frontline health care providers, EMS staff members are in a pivotal position to recognize and effectively manage the mental health needs of patients and their families. CONCLUSIONS: Ecological changes in emergency departments, such as linkages to mental health follow-up services, training of EMS providers and mental health professionals, and focused research that provides an empirical basis for practice, are necessary components for improving current standards of health care. PMID- 11733301 TI - Ethics seminars: HIV testing, consent, and physician responsibilities. AB - Emergency physicians constantly have multiple ethical obligations in the emergency department. They must understand these sometimes conflicting obligations and learn to prioritize. A case discussion is presented that exemplifies the conflict between patient privacy and society's right to know. Specific aspects of HIV testing and obtaining patient consent are presented. Teaching physicians are encouraged to use such common cases for "ethics case discussion." PMID- 11733302 TI - Emergency medicine resident patient care documentation using a hand-held computerized device. AB - OBJECTIVE: To determine whether emergency medicine (EM) resident documentation of procedures, patient encounters, and patient follow-ups improved after implementation of a personal digital assistant (PDA) hand-held recording system. METHODS: All first-year EM residents were provided a PalmV (Palm, Inc., Santa Clara, CA) PDA. A customized patient procedure and encounter program was constructed using Pendragon Forms (Pendragon Software Corporation, Libertyville, IL) and loaded into each PDA. Residents were instructed to enter information on patients who had any of 21 procedures performed or were considered to be clinically unstable. These data were downloaded to the residency coordinator's desktop computer. The mean number of procedures, encounters, and follow-ups performed per resident were then compared with those of a group of 36 historical controls from the three previous first-year resident classes who recorded the same information using a handwritten card system. Data from the historical controls were combined and the means of each group were compared by Student's t test. RESULTS: Mean documentation of three procedures was significantly increased in the PDA group versus the index card system: conscious sedation 5.8 vs. 0.03 (p < 0.000005), thoracentesis 2.2 vs. 0.0 (p = 0.002), ultrasound 6.3 vs. 0.0 (p = 0.002). The mean numbers of pericardiocenteses and unstable pediatric surgical patient evaluations were significantly decreased in the hand-held group [from 1.2 to 0.4 (p = 0.03) and from 9.1 to 2.2 (p = 0.02), respectively]. Patient follow up documentations were not statistically different between the two groups. CONCLUSIONS: Use of a hand-held PDA was associated with an increase in first-year EM resident documentation in three of 20 procedures and a decrease in one procedure and the number of unstable surgical pediatric patient resuscitations. The overall time savings in constructing a resident procedure database, as well as the other uses of the PDAs, may make transition to a hand-held computer-based procedure log an attractive option for EM residencies. PMID- 11733305 TI - Technical aspects of neuroangiography: are risks and safeguards understood in the same way? PMID- 11733306 TI - Device-mediated effect versus the healing response in interventional neuroradiology. PMID- 11733307 TI - Functional MR imaging evaluation of spinal cord function by use of neurophysiological stimuli. PMID- 11733308 TI - Pictorial review of glutamate excitotoxicity: fundamental concepts for neuroimaging. PMID- 11733309 TI - Aneurysmal rupture during embolization with Guglielmi detachable coils: causes, management, and outcome. AB - BACKGROUND AND PURPOSE: Aneurysmal rupture during endovascular treatment is one of the most feared complications of endovascular aneurysm therapy. The purpose of this study was to determine the frequency, causes, management, and outcome of aneurysmal rupture that occurred during treatment with Guglielmi detachable coils (GDCs) in an unselected series of patients with ruptured cerebral aneurysms. METHODS: Between July 1997 and December 2000, we treated 164 acutely ruptured cerebral aneurysms with GDCs. All charts were reviewed, and patients with aneurysmal rupture occurring during embolization were identified. RESULTS: Five patients had an intraprocedural aneurysmal rupture. In one patient, rupture was due to guidewire perforation of the wall. In two patients, the microcatheter itself perforated the aneurysm. In another two patients, rupture occurred during placement of the first coil. Endovascular packing was continued in all patients. One patient died as a result of the aneurysmal rupture. No negative long-term effects were observed in the remaining four patients. In summary, we observed intraprocedural aneurysmal rupture in 3% of our patients, with a mortality rate of 20% and no long-term morbidity. CONCLUSION: Aneurysmal rupture during endovascular treatment with GDCs is a rare event; clinical severity may be variable. Embolization of the aneurysm can be continued in most cases, and most patients with treatment-related subarachnoid hemorrhage survive without serious sequelae. PMID- 11733310 TI - Elastase-induced saccular aneurysms in rabbits: comparison of geometric features with those of human aneurysms. AB - BACKGROUND AND PURPOSE: The development of more effective intracranial aneurysm therapy depends on the ability to test various intravascular occlusion devices and techniques in preclinical animal models. This requires the creation of experimental aneurysms, which, ideally, should mimic the size and geometric features of human intracranial aneurysms. The purpose of this study was to characterize the morphologic features of elastase-induced saccular aneurysms in rabbits to determine whether the morphology of such aneurysms mimics that of human intracranial aneurysms. METHODS: Elastase-induced saccular aneurysms were created in 40 New Zealand white rabbits. Intravenous digital subtraction angiography was performed 14 days after surgery. Relative to an external sizing device, the following dimensions were determined: aneurysm dome (height and width), aneurysm neck diameter, and parent artery diameter. Based on maximal diameter, aneurysms were categorized as small (2.0-4.9 mm), medium-sized (5.0-9.9 mm), or large (10-16 mm), and as narrow-necked (<4.0 mm neck width) or wide necked (>4.0 mm neck width). Mean dome-neck ratio was calculated and compared with that of human aneurysms. RESULTS: All aneurysm cavities were angiographically patent. Widths of the cavities ranged from 2.5 to 7.1 mm (mean, 4.1 +/- 1.2 mm); heights ranged from 3.0 to 15.6 mm (mean, 8.8 +/- 2.6 mm). Three (7.5%) of 40 aneurysms were small, 20 (50%) were medium-sized, and 17 (42.5%) were large. Twenty-two (55%) of 40 aneurysms were small-necked, and 18 (45%) were wide-necked. Mean dome-neck ratio was 1.13 +/- 0.54. Mean parent artery diameter was 4.3 +/- 1.4 mm. CONCLUSION: Saccular aneurysms of sizes similar to that of human intracranial aneurysms were reliably created using a simple method of vessel ligation and elastase injury. Neck sizes varied with both large and small necked aneurysms created. PMID- 11733311 TI - Injection rates for neuroangiography: results of a survey. AB - BACKGROUND AND PURPOSE: Injection rates have attracted scrutiny because of an incident of an aneurysm rupturing during arteriography. We sought to determine the current injection rates for neuroangiography in the setting of aneurysm evaluation. METHODS: An e-mail survey was distributed to 90 neuroradiology program directors within the United States and Canada. The injection rates and total volumes of contrast material injected for the common carotid, internal carotid, and vertebral arteries were provided for an "average" adult individual evaluated for intracranial aneurysms. RESULTS: Sixty-three (70.0%) program directors replied to the survey. Of these, five perform hand injections only and provided approximate values. The mean injection rates (SD) and total volumes (SD) for common carotid arteries were 7.2 cm(3)/s (1.8) and 9.9 cm(3) (2.0), respectively; for internal carotid arteries, 5.8 cm(3)/s (1.4) and 7.9 cm(3) (1.5); and for vertebral arteries, 5.4 cm(3)/s (1.2) and 7.8 cm(3) (1.7). The modes (rate/total) for the common carotid, internal carotid, and vertebral arteries were 7/12, 6/8, and 5/8, respectively. Forty-eight (81.4%) of 59 respondents did not believe a reduction in current injection rates would lead to a diminution in complications of arteriography. CONCLUSION: The rates of injection of contrast material in the United States for neuroradiologic studies show great variability. It does not seem that reducing arteriographic complications is an impetus to reduce injection rates. The values in this survey can provide "industry norms" for injections in the common carotid, internal carotid, and vertebral arteries if these rates are challenged. PMID- 11733312 TI - Delayed stroke secondary to increasing mass effect after endovascular treatment of a giant aneurysm by parent vessel occlusion. AB - A 47-year-old woman, who had lost vision in her left eye because of a giant left supraclinoid internal carotid artery aneurysm, was referred for endovascular treatment. Parent-vessel occlusion was performed to obtain circulatory exclusion of the aneurysm. Eight days after treatment, she became hemiparetic and dysphasic. Repeat angiography showed compression of the left middle cerebral artery by the swelling giant aneurysm. Preventive measures should be taken to avert worsening of mass effect when giant aneurysms become thrombotic. PMID- 11733313 TI - Treatment of a ruptured dissecting vertebral artery aneurysm with double stent placement: case report. AB - A ruptured dissecting right vertebral artery aneurysm was treated by means of double stent placement with two overlapping stents. Control angiography performed 3 d after stent placement revealed beginning aneurysmal thrombosis. Substantial reduction in aneurysmal size was observed after 4 wk, whereas total occlusion was observed after 3 mo. The reduced stent porosity caused by the overlapping stents, which result in significant hemodynamic changes inside the aneurysmal sac, may accelerate intraaneurysmal thrombosis and may be helpful in achieving a more rapid complete occlusion compared with that achieved by single stent placement. PMID- 11733314 TI - Carbon dioxide column angioscopy: a new endovascular imaging technique. AB - A new angioscopic technique with a CO(2) gas medium for prolonged viewing sessions in the carotid artery is described. A stationary column of CO(2) gas, angled 17-30 degrees subhorizontally and buoyed against a balloon catheter, can be safely maintained. During 10-20-min sessions in dogs, endothelia, thrombi, stent filaments, coils, and an intimal flap were visualized. This technique eliminates the need for continuous saline infusion, which has prevented the application of angioscopy in the carotid artery. PMID- 11733315 TI - Functional MR imaging of the cervical spinal cord by use of median nerve stimulation and fist clenching. AB - BACKGROUND AND PURPOSE: Findings of blood oxygen level-dependent (BOLD) functional MR (fMR) imaging of the cervical spinal cord, obtained by using a fist clenching motor task, have been sporadically reported. Because spinal activation by sensory stimuli has a potential at least equal to that of fist clenching, its feasibility was assessed. Whether stimulation of the median nerve could evoke an fMR imaging response at 1.5 T in the cervical spinal cord was investigated, and the response pattern was compared with that obtained by fist clenching. METHODS: A dynamic cardiac-gated T2*-weighted imaging sequence was used to quantify cervical spinal cord activation under two paradigms with different numbers of subjects. Seven subjects underwent electrical median nerve stimulation at the elbow sufficient to elicit a maximal compound muscle action potential in the flexor carpi radialis muscle. Eleven subjects performed self-paced fist clenching. Cord activation was measured in the sagittal and transverse imaging planes. RESULTS: In the sagittal view, five of seven subjects had an fMR imaging response in the lower cervical spinal cord upon median nerve stimulation, whereas seven of 11 subjects showed activation with the fist-clenching task. Within the cord, the measured fMR imaging response level was approximately 8-15% with respect to the baseline signal level. In the transverse imaging plane, significant fMR imaging responses could be measured in only two of six and six of nine subjects with median nerve stimulation or fist clenching, respectively. A consistent cross-sectional localization of the activity measured in the spinal cord was not detected, either in terms of the right and left sides or in terms of the posterior and anterior directions. CONCLUSION: In the sagittal plane, median nerve stimulation at the elbow can evoke an fMR imaging response in the lower cervical spinal cord. The activation pattern was comparable with that obtained by fist clenching. The localization of the segmental fMR imaging activation (C4 through T1) is consistent with the known functional neuroanatomy for both paradigms. In the transverse plane, reliable fMR imaging responses were obtained much less frequently, and assignment of distinct areas of the spinal cord to the stimulation methods used was not possible. PMID- 11733317 TI - MR findings of a primary intramedullary malignant melanoma: case report and literature review. AB - A case of primary malignant melanoma of the conus medullaris depicted at MR imaging is presented. Tumoral histoimmunologic analysis revealed features of malignant melanoma. Because findings for primary melanoma outside the spinal cord were negative, the diagnosis of primary intramedullary malignant melanoma was established. This rare tumor should be suspected when T1-weighted images show signal hyperintensity and T2-weighted images show signal iso- or hypointensity, with mild contrast enhancement of the lesion. However, these features may vary depending on intratumoral bleeding and melanin content. PMID- 11733316 TI - Age of fracture and clinical outcomes of percutaneous vertebroplasty. AB - BACKGROUND AND PURPOSE: The patient populations that are most likely to benefit from percutaneous vertebroplasty (PVP) are uncertain. Our purpose was to evaluate the effect of the age of vertebral compression fracture (VCF) on clinical improvement after PVP. METHODS: We performed a retrospective review of charts of patients who had undergone PVP for painful osteoporotic VCFs at our institution. The preprocedural and postprocedural outcome measurements of pain, mobility, and analgesic use were compared for 80 treatment sessions in 75 patients (122 total vertebrae treated). We assessed the association between the duration of pain before PVP and postprocedural outcomes by using multivariable analysis. RESULTS: Age of fracture at time of PVP was not independently associated with postprocedural pain or activity. Increasing age of fracture was independently associated with slightly greater postprocedural analgesic requirement, at least for patients who required narcotics at baseline before PVP. Greater preprocedural analgesic requirement was independently associated with greater postprocedural analgesic requirement. Reduced preprocedural mobility was independently associated with reduced postprocedural mobility. CONCLUSION: PVP is a highly efficacious therapy for relief of pain and improvement in mobility, regardless of fracture age. PVP also is efficacious in reducing analgesic requirement, although this effect may be slightly blunted in patients who require narcotics before the procedure and in those who have older fractures. PMID- 11733318 TI - Petrous apex cephaloceles. AB - BACKGROUND AND PURPOSE: Petrous apex cephaloceles (PACs) are uncommon lesions that are usually incidental but may be symptomatic. We reviewed MR and CT studies in 10 patients with PACs to identify characteristic imaging features that facilitate their diagnosis. METHODS: MR and CT studies from 10 patients with PACs were reviewed retrospectively. In each case the PAC was characterized by lesion center, signal intensity or attenuation, adjacent petrous apex pneumatization, and its relationship to Meckel's cave. Intraoperative findings were reviewed in the three cases in which surgery was performed. RESULTS: All 10 patients had lobulated expansile cystic petrous apex lesions centered along the posterolateral margin of Meckel's cave. All cysts were contiguous with Meckel's cave. Three patients had bilateral PACs. Four patients had symptoms that could potentially be explained by the PAC, while findings in the other six were incidental observations. Three patients underwent surgery, during which two lesions were diagnosed as meningoceles while the third was diagnosed as an arachnoid cyst protruding through a dural defect. CONCLUSION: PACs represent a protrusion of meninges and CSF from the posterolateral portion of Meckel's cave into the petrous apex, which is their characteristic imaging appearance. PACs are usually incidental but may be symptomatic. Surgical intervention should be approached cautiously and undertaken only when symptoms are clearly linked to the presence of this lesion. PMID- 11733319 TI - Normal laryngeal CT findings after supracricoid partial laryngectomy. AB - BACKGROUND AND PURPOSE: Supracricoid horizontal partial laryngectomy (SCPL) is increasingly used to treat endolaryngeal carcinoma. However, few radiologic reports of these procedures exist. Our purpose was to evaluate the normal CT appearance of the neolarynx after surgery. METHODS: SCPL includes cricohyoidopexy (CHP), cricohyoidoepiglottopexy (CHEP), and tracheocricohyoidoepiglottopexy (TCHEP). We examined CT scans obtained from 18 patients without local superficial recurrence who underwent SCPL: 10, CHEP; seven, CHP; and one, TCHEP. Three reference sections were used to analyze the main surgical reconstruction: an upper section through the hyoid bone, a lower section through the cricoid cartilage, and a middle section in between. The distance between the hyoid bone and cricoid cartilage was measured. RESULTS: The epiglottis and valleculae were visible in the upper section in seven of 10 patients who underwent CHEP; this finding allowed distinction between CHEP and CHP. The arytenoids were depicted in 13 of 18 cases and reflected neolaryngeal shortening. The lower section showed the empty cricoid lumen lined by a thin mucosa; the anterior arch of the cricoid was amputated at TCHEP. The middle section showed the neovestibule, the lateral boundaries of which were the hypertrophic neoaryepiglottic folds; the anterior limit was the epiglottis for CHEP or the base of the tongue for CHP. The average distance between the hyoid bone and cricoid cartilage was 11 mm. CONCLUSION: Normal CT anatomy of the larynx after SCPL is defined. Three key sections may accurately distinguish the various types of SCPL. CT is a valuable tool for depicting tumor recurrence, especially when the tumor is submucosal. PMID- 11733320 TI - High-sensitivity coil array for head and neck imaging: technical note. AB - The purpose of this study was to develop coils for MR imaging of the head and neck region, with the aim of improving sensitivity and coverage. A head and neck phased array coil was constructed and compared with volume and temporomandibular joint surface coils for sensitivity and coverage in phantom studies. An algorithm was implemented to correct for the nonuniformity in the surface coil reception profile. Its application to high-resolution T2-weighted imaging in healthy volunteers was investigated. PMID- 11733321 TI - Unusual CT appearance in an odontogenic keratocyst of the mandible: case report. AB - An expansile lesion in the body of the left mandible had high attenuation (225 HU) on nonenhanced CT scans. Histologic examination revealed an odontogenic keratocyst with no evidence of mineralization or calcification within the lesion. The high attenuation was considered to be due to highly concentrated protein of thick, viscous keratin in the lumen of the keratocyst. PMID- 11733322 TI - Solitary fibrous tumor in the buccal space: MR findings with pathologic correlation. AB - We report MR findings in a case of a solitary fibrous tumor involving the buccal space in a middle-aged man. On MR images, most of the mass was isointense and hyperintense to the muscle on T1- and T2-weighted images, respectively, and showed homogeneously strong enhancement on contrast-enhanced T1-weighted images. The medial peripheral portion, which was isointense on T2-weighted images and showed less enhancement on contrast-enhanced T1-weighted images, corresponded to the hypocellular and collagenous sclerotic area on pathologic correlation. PMID- 11733323 TI - Analysis of normal-appearing white matter in multiple sclerosis: comparison of diffusion tensor MR imaging and magnetization transfer imaging. AB - BACKGROUND AND PURPOSE: Our purpose was to compare diffusion tensor MR and magnetization transfer imaging in assessing normal-appearing white matter (WM) regions in multiple sclerosis (MS). METHODS: Diffusion tensor, magnetization transfer, and conventional MR imaging were performed in 12 patients with MS. Fractional anisotropy, apparent diffusion coefficients (ADCs), and magnetization transfer ratios (MTRs) were measured in plaques, normal-appearing periplaque WM (PWM) regions, and normal-appearing WM regions remote from plaques. Mean fractional anisotropy, ADCs, and MTRs were calculated and compared in WM regions. RESULTS: Fractional anisotropy was lower in normal-appearing PWM regions than in remote WM regions (P <.001) but higher than in plaques (P <.001). MTRs were lower (not significantly, P =.19) in normal-appearing PWM regions than in remote regions. MTRs were higher in normal-appearing PWM regions than in plaques (P <.001). ADCs were higher in normal-appearing PWM regions than in remote regions (P =.008) but lower than in plaques (P =.001). Correlation between fractional anisotropy and MTRs of individual lesions was poor (r = 0.18) and between fractional anisotropy and ADC, modest (r = -0.39). CONCLUSION: In MS, diffusion tensor MR imaging can depict differences between WM regions that are not apparent on conventional MR images. Anisotropy measurements may be more sensitive than those of MTRs in detecting subtle abnormalities in PWM. PMID- 11733324 TI - Etiology of cortical and white matter lesions in cyclosporin-A and FK-506 neurotoxicity. AB - BACKGROUND AND PURPOSE: The etiology of the neurotoxicity associated with cyclosporin-A (CsA) and FK-506 treatment is not fully understood. At our institution, we noticed a distinct, abrupt change in the imaging characteristics of CsA and FK-506 neurotoxicity, which consisted of a shift in lesion morphology from a white matter abnormality to a mixed cortical and white matter pattern. The purpose of this study was to assess clinical parameters that might explain this change. METHODS: Twenty-two patients had a neurotoxic reaction and brain imaging changes while receiving CsA or FK-506. Nineteen patients received allogeneic bone marrow transplants, and three had aplastic marrow disorders. Fifty-one imaging studies (CT or MR imaging) were obtained, and lesion characteristics, locations, and time courses were evaluated along with relevant clinical data. RESULTS: Nine patients who had been conditioned for transplantation with cyclophosphamide and chemotherapy (busulfan or thiotepa) had a mixed pattern of cortical and white matter involvement (57 lesions). Isolated white matter involvement (62 lesions) developed in three nontransplant patients and 10 transplant patients conditioned with cyclophosphamide and total-body irradiation. All lesions occurred at typical brain watershed zones. Lesion enhancement was noted in two patients conditioned with chemotherapy. Initial images demonstrated characteristic lesions in 15 patients (68%). Initial images were normal in four patients (18%) and nonspecific in three patients (14%). CONCLUSION: Lesion location in CsA and FK-506 neurotoxicity may depend on the presence or type of conditioning used before bone marrow transplantation. Nontransplant patients or those conditioned with total body irradiation develop white matter lesions, whereas those conditioned with chemotherapy develop mixed cortical and white matter lesions. PMID- 11733325 TI - Blood volume of gliomas determined by double-echo dynamic perfusion-weighted MR imaging: a preliminary study. AB - BACKGROUND AND PURPOSE: After bolus injection, gadopentetate dimeglumine causes a T2* rate change in permeable tissue that is contaminated by the T1 shortening effect due to the leakage of contrast agent. Therefore, tumor vascularity as reported in previous single-echo perfusion-weighted MR imaging studies has been underestimated. Our aim was to quantitatively and qualitatively evaluate the degree of blood volume of glioblastoma multiformes (GBMs) underestimated by this T1 shortening effect. METHODS: We used double-echo dynamic MR imaging after a bolus injection of gadopentetate dimeglumine (double-echo perfusion-weighted MR imaging) to simultaneously determine tumor blood volume without (V(T1U)) and with (V(T1C)) T1 shortening correction. MR imaging was performed in five consecutive patients with GBMs. The ratios of V(T1U) and V(T1C) were calculated and compared by means of quantitative analysis. The degree of tumor blood volume as determined by V(T1U) and V(T1C) maps were qualitatively compared using a three-point scale. RESULTS: All GBMs showed contrast enhancement on postcontrast T1-weighted images. In all subjects, the values of V(T1U) were significantly lower than those of V(T1C) (mean +/- SD, 2.05 +/- 1.01 vs. 3.62 +/- 1.40, respectively [P <.05]), indicating that tumor blood volume obtained by double-echo perfusion-weighted MR imaging was significantly higher than that by single-echo imaging. In the qualitative analysis, tumor blood volume on the V(T1U) map was less conspicuous than that on the V(T1C) map. CONCLUSION: Careful attention should be paid to the underestimation of tumor blood volume resulting from T1 shortening effects when using single-echo perfusion-weighted MR imaging. Double-echo imaging may be more suitable for the analysis of blood volume in GBMs. PMID- 11733326 TI - Proton MR spectroscopy of cortical tubers in adults with tuberous sclerosis complex. AB - BACKGROUND AND PURPOSE: Significant advances have been made in understanding the origin of brain manifestations associated with tuberous sclerosis complex (TSC), most recently the idea that cortical tubers and subependymal nodules are a disorder of neocortical formation. The present study was designed to test the hypothesis that MR proton spectra of cortical tubers are abnormal because of the developmental immaturity of the tubers' neurons and glia. METHODS: Twenty-six adults with TSC were studied. Proton spectroscopy was performed with a single voxel point-resolved spectroscopy technique in two 8-mL regions: one over a cortical tuber and the other over a corresponding anatomic area of normal appearing brain in the contralateral cerebral hemisphere. The results were expressed in terms of the ratio of the area under the three prominent resonances of choline (Cho), creatine + phosphocreatine (Cr), and N-acetyl groups (NA). RESULTS: Cortical tubers showed statistically significantly lower NA/Cr (1.54 +/- 0.24 vs. 1.72 +/- 0.23), NA/(Cho+Cr) (0.80 +/- 0.14 vs. 0.91 +/- 0.12), and NA/Cho (1.69 +/- 0.36 vs. 1.97 +/- 0.38) than did normal-appearing brain. No statistically significant difference was found in the Cho/Cr ratios between the tubers and the contralateral brain (0.93 +/- 0.15 vs. 0.90 +/- 0.12). No lactate was identified in any cortical tubers. CONCLUSION: Proton spectroscopy can show differences between cortical tubers and normal-appearing white matter in patients with TSC. The reduced NA/Cr ratio is probably due to reduced levels of NA, most likely caused by the presence of immature neurons and glia in tubers that do not express NA, or by the presence of gliosis, a known histologic component of tubers. PMID- 11733327 TI - Sequential MR imaging and proton MR spectroscopy in patients who underwent recent detoxification for chronic alcoholism: correlation with clinical and neuropsychological data. AB - BACKGROUND AND PURPOSE: Chronic alcohol abuse may cause neuropsychological disorders and result in brain atrophy. The purpose of this study was to evaluate the metabolic, morphologic, and functional cerebral changes in the early stage of abstinence from chronic alcoholism. METHODS: Seventeen alcohol-dependent patients underwent MR imaging and MR spectroscopy on days 1 through 3 and days 36 through 39 of abstinence. In addition, psychological performance measures testing intelligence, concentration, attention, and memory were applied. Neuropsychological data were correlated with spectroscopic and volumetric results by using a Pearson's product moment correlation. The same measurements were also performed in 12 healthy, age-matched control subjects. Peak integral values for N acetylaspartate (NAA) and choline (Cho) were referred to the peak integral value of creatine (Cr) as the internal reference. RESULTS: NAA/Cr was decreased in the patients in both the frontal lobes and cerebellum immediately after cessation of drinking (days 1 through 3). After 36 to 39 days of abstinence, NAA/Cr had significantly increased in the patients and corresponded to performance on psychological tests. The Cho/Cr ratio was decreased in the cerebellum during early abstinence but was recovered on days 36 through 39. The patients had enlarged CSF spaces 1 to 3 days after detoxification, which decreased during sobriety. The extent of brain atrophy did not correspond to performance on psychological performance tests. CONCLUSION: Regression of brain atrophy and metabolic recovery occurs at an early stage after abstinence from chronic alcohol abuse. MR spectroscopy findings return to normal metabolic levels within weeks after detoxification. The recovery of NAA/Cr is associated with improved performance on neuropsychological tests. PMID- 11733328 TI - In vivo detection of postictal perturbations of cerebral metabolism by use of proton MR spectroscopy: preliminary results in a canine model of prolonged generalized seizures. AB - BACKGROUND AND PURPOSE: Biochemical studies of seizures in patients and laboratory animals have monitored postictal perturbations in cerebral metabolism with either invasive techniques or with such noninvasive techniques as nuclear medicine, MR imaging, in vivo phosphorus MR spectroscopy (MRS), and in vivo proton MRS at field strengths of 1.5 T or above. We investigated postictal metabolic changes in a generalized seizure model with in vivo proton MRS at 0.5 T, in which the combination of glutamate and glutamine resonances (denoted glx) can be modeled as a singlet. METHODS: Five adult mongrel dogs underwent control and postictal experiments in which single-voxel proton MR spectra were obtained from the right frontal lobe cortex with a point-resolved spectroscopy technique approximately every 20 minutes for 3 hours. N-acetylaspartate (NAA), glx, and creatine (Cr) were quantified in absolute millimolar units with a cerebral water referenced algorithm. Inter- and intrasubject differences in mean metabolite concentrations collected throughout the 3-hour period were compared using an unpaired, two-tailed Student's t test at a.05 level of significance. RESULTS: We found a significant increase (15.4%) in the postictal intersubject mean glx concentration, as well as a 23.7% postictal decrease in the intersubject mean Cr concentration. A trend toward a subtle decrease in postictal intersubject mean NAA concentration was not statistically significant. We also observed a substantial qualitative increase in the combination of postictal lactate and free fatty acid peaks. CONCLUSIONS: The glx, NAA, lactate, and free fatty acid results are in general agreement with previous studies of postictal perturbations in cerebral metabolism measured with invasive biochemical or noninvasive high-field strength in vivo MRS detection assays. Given a high sensitivity for glx at 0.5 T relative to 1.5 T, further studies of postictal mesial temporal lobe structures are warranted in chronic animal preparations that model temporal lobe epilepsy. PMID- 11733329 TI - Focal enhancement of cranial nerve V after radiosurgery with the Leksell gamma knife: experience in 15 patients with medically refractory trigeminal neuralgia. AB - BACKGROUND AND PURPOSE: Gamma knife radiosurgery is an alternative for the treatment of medically refractory trigeminal neuralgia. Few reports of posttreatment MR imaging appearance of cranial nerve V exist. Our purpose was to define MR imaging characteristics in cranial nerve V after gamma knife radiosurgery. METHODS: We retrospectively reviewed MR images of 15 patients who underwent gamma knife radiosurgery for trigeminal neuralgia. Radiation doses were 35-45 Gy at the 50% isodose line. Thin-section T2-weighted images and contrast enhanced and nonenhanced T1-weighted images were obtained the day of radiosurgery and within the next 5 mo. Images were scored for enhancement and hyperintensity on T2-weighted images. Time to follow-up imaging and radiation dose were recorded. RESULTS: Mean time to follow-up imaging was 61 +/- 29 d. Posttreatment T2-weighted images showed stable signal intensity in all cases, with radiosurgical target site enhancement in 10. All five patients whose images did not show treatment-related enhancement received radiation doses of 35 Gy. The data suggested a correlation between enhancement with radiation dose (P =.06). No correlation of enhancement with treatment response or time to follow-up existed (P >.05). CONCLUSION: The trigeminal nerve often enhances at the target site after radiosurgery. Lack of trigeminal nerve enhancement occurred only with lower doses (35 Gy at 50%). MR imaging may be useful to confirm the presence and location of the treatment site after gamma knife radiosurgery for trigeminal neuralgia. PMID- 11733330 TI - Diffusion-weighted MR imaging of an acute venous stroke: case report. AB - A patient with a superior sagittal sinus thrombosis had progressively worsening symptoms and signs that resolved after IV heparin therapy. MR imaging revealed abnormalities in diffusion, similar to those seen with acute arterial stroke. Abnormalities shown on a T2-weighted fast spin-echo and fluid-attenuated inversion recovery images resolved completely. The findings in this report contradict those from previous reports that suggest diffusion-weighted imaging with quantitative apparent diffusion coefficients may be used in selecting patients for dural venous sinus thrombolysis. PMID- 11733331 TI - Congenital absence of the internal carotid artery: case reports and review of the collateral circulation. AB - Absence of the internal carotid artery (ICA) is a rare congenital anomaly. The embryology of the ICA and the common collateral pathways associated with its congenital absence are reviewed, with four new cases provided for illustration. While collateral blood flow may allow these patients to remain asymptomatic, two of our patients presented with transient ischemic attacks. Recognition of this anomaly has important implications during planned carotid or transsphenoidal surgery, in thromboembolic disease, and in the surveillance and detection of associated cerebral aneurysms. PMID- 11733332 TI - Intradural primary chondroblastic osteosarcoma: case report. AB - We report a rare case of intradural primary osteosarcoma (IPOS) in a 74-year-old man with aphasia and right-sided hemiparesis. Radiologic workup revealed a large, partially calcified, left-sided frontotemporal intracranial mass lesion. At surgery, the tumor was found to be entirely intradural; it involved the brain and subarachnoid space of the left sylvian fissure. The adjacent dura was uninvolved. Neuropathologic findings confirmed the diagnosis of chondroblastic osteosarcoma. To our knowledge, this is the sixth reported case of IPOS and the first reported case of the chondroblastic subtype. PMID- 11733333 TI - Optic pathway glioma: correlation of imaging findings with the presence of neurofibromatosis. AB - BACKGROUND AND PURPOSE: Despite the benign histology of optic pathway glioma (OPG) (low-grade astrocytoma), its biological behavior is unpredictable, and it is unclear whether specific morphologic or anatomic patterns may be predictive of prognosis. It is also unclear whether OPG associated with neurofibromatosis (NF) is a distinct entity from non-NF-OPG. Our purpose was to describe the MR imaging features of OPG, compare the findings between patients with and those without NF, and identify prognostic imaging signs. METHODS: MR examinations of 91 patients with OPG (47 with NF and 44 without) were reviewed at presentation and during follow-up. The images were evaluated for size and extension of tumor, and imaging parameters. Statistical bivariate analysis was used to compare the patients with and those without NF, and Pearson correlation was used to evaluate the correlation between the different imaging parameters and prognosis. Kappa values were calculated to determine intraobserver and interobserver variability. RESULTS: The most common site of involvement in the NF group was the orbital nerve (66%), followed by the chiasm (62%). In the non-NF group, the chiasm was the most common site of involvement (91%); the orbital nerves were involved in only 32%. Extension beyond the optic pathway at diagnosis was uncommon in the NF group (2%) but frequent in the non-NF group (68%). In the NF group, the tumor was smaller and the original shape of the optic pathways was preserved (91% vs. 27% in the non-NF group). The presence of cystic components was significantly more common in the non-NF patients (66% vs. 9% in the NF group). During follow-up, half the NF patients remained stable, in contrast to 5% of the non-NF group. No statistical correlation was found between imaging features and biological behavior of the tumor. CONCLUSION: NF-OPG is a separate entity from non-NF-OPG, with different imaging features and prognosis, thereby warranting a specific diagnostic, clinical, and therapeutic approach. PMID- 11733334 TI - Unusual imaging appearance of an intracranial dermoid cyst. AB - Intracranial dermoid cysts have characteristic CT and MR imaging findings that generally make preoperative diagnosis straightforward. Enhancement of uncomplicated intradural dermoid cysts on CT or MR studies has been reported but is rare. We present a case of a posterior fossa dermoid cyst that was not only hyperattenuating on CT scans but also contained a mural nodule with clear evidence of enhancement on MR images. PMID- 11733337 TI - Thirty-ninth annual meeting of the American Society of Neuroradiology. PMID- 11733339 TI - Thallum-201 single-proton emission CT versus CT for the detection of recurrent squamous cell carcinoma of the head and neck. PMID- 11733340 TI - Spinal cord involvement in CNS Whipple disease: an ongoing experience in the magnetic resonance imaging era. PMID- 11733341 TI - MR images of postmortem fetuses. PMID- 11733346 TI - Translocations in malignant tumors. PMID- 11733347 TI - Aluminum chloride pretreatment of elastin inhibits elastolysis by matrix metalloproteinases and leads to inhibition of elastin-oriented calcification. AB - Calcification of elastin occurs in many pathological cardiovascular diseases including atherosclerosis. We have previously shown that purified elastin when subdermally implanted in rats undergoes severe calcification and aluminum chloride (AlCl(3)) pretreatment of elastin inhibits calcification. In the present study we investigated whether matrix metalloproteinase (MMP) binding to elastin and elastin degradation is prevented by AlCl(3) pretreatment. Subdermal implantation of AlCl(3)-pretreated elastin showed significantly lower MMP-9 and MMP-2 activity surrounding the implant as compared to the control implants. AlCl(3) pretreatment also significantly inhibited elastin implant calcification at the seven-day implant period (AlCl(3)-pretreated 4.07 +/- 1.27, control 23.82 +/- 2.24 microg/mg; p<0.0001). Moreover, elastin gel zymography studies showed that gel pretreatment with AlCl(3) inhibited elastolysis by MMP-9. We also demonstrate significant suppression of MMP-2 activity in aortic wall segments of AlCl(3)-pretreated porcine bioprosthetic heart valve implants as compared to control valve implants in sheep mitral valve replacement studies. AlCl(3) pretreatment also significantly inhibited calcification of elastin in this model. Thus, we conclude that aluminum ion binding to elastin prevents MMP-mediated elastolysis and thus prevents elastin calcification. PMID- 11733348 TI - BRD4 bromodomain gene rearrangement in aggressive carcinoma with translocation t(15;19). AB - Translocation t(15;19)(q13;p13.1) defines a lethal midline carcinoma arising adjacent to respiratory tract in young people. To characterize molecular alterations responsible for the distinctly aggressive biological behavior of this cancer, we mapped the chromosome 15 and 19 translocation breakpoints by fluorescence in situ hybridization (FISH) and Southern blotting. To evaluate preliminarily the frequency, anatomical distribution, and histological features of t(15;19) cancer, we developed a FISH assay for paraffin sections. Our findings reveal a novel oncogenic mechanism in which the chromosome 19 translocation breakpoint interrupts the coding sequence of a bromodomain gene, BRD4. These studies implicate BRD4 as a potential partner in a t(15;19)-associated fusion oncogene. In addition, we localized the chromosome 15 breakpoint to a 9-kb region in each of two cases, thereby identifying several candidate oncogenes which might represent the BRD4 fusion partner. FISH evaluation of 13 pediatric carcinomas revealed t(15;19) in one of four sinonasal carcinomas, whereas this translocation was not detected in thymic (n = 3), mucoepidermoid (n = 3), laryngeal (n = 2), or nasopharyngeal (n = 1) carcinomas. Our studies shed light on the oncogenic mechanism underlying t(15;19) and provide further evidence that this highly lethal cancer arises from respiratory mucosa. PMID- 11733349 TI - Deposition of transthyretin in early stages of familial amyloidotic polyneuropathy: evidence for toxicity of nonfibrillar aggregates. AB - Familial amyloidotic polyneuropathy (FAP) is a neurodegenerative disorder characterized by extracellular deposition of transthyretin (TTR) amyloid fibrils, particularly in the peripheral nervous system. No systematic immunohistochemical data exists relating TTR deposition with FAP progression. We assessed nerves from FAP patients in different stages of disease progression (FAP 0 to FAP 3) for TTR deposition by immunohistochemistry, and for the presence of amyloid fibrils by Congo Red staining. The nature of the deposited material was further studied by electron microscopy. We observed that early in FAP (FAP 0), TTR is already deposited in an aggregated nonfibrillar form, negative by Congo Red staining. This suggested that in vivo, preamyloidogenic forms of TTR exist in the nerve, in a stage before fibril formation. Cytotoxicity of nonfibrillar TTR was assessed in nerves of different FAP stages by immunohistochemistry for macrophage colony stimulating factor. FAP 0 patients already presented increased axonal expression of macrophage colony-stimulating factor that was maintained in all other stages, in sites related to TTR deposition. Toxicity of synthetic TTR fibrils formed in vitro at physiological pH was studied on a Schwannoma cell line by caspase-3 activation assays and showed that early aggregates but not mature fibrils are toxic to cells. Taken together, these results show that nonfibrillar cytotoxic deposits occur in early stages of FAP. PMID- 11733350 TI - Neutrophil transmigration in inflammatory bowel disease is associated with differential expression of epithelial intercellular junction proteins. AB - Inflammatory bowel disease (IBD) consisting of ulcerative colitis (UC) and Crohn's (CD) typically displays a waxing and waning course punctuated by disease flares that are characterized by transepithelial migration of neutrophils (PMN) and altered barrier function. Since epithelial barrier function is primarily regulated by the apical most intercellular junction referred to as the tight junction (TJ), our aim was to examine expression of TJ and adherens junction (AJ) proteins in relation to PMN infiltration in mucosal tissue samples from patients with active IBD. Expression of epithelial intercellular TJ proteins (occludin, ZO 1, claudin-1, and JAM) and subjacent AJ (beta-catenin and E-cadherin) proteins were examined by immunoflourescence/confocal microscopy, immunohistochemistry, and Western blotting. Colonic mucosa from patients with UC revealed dramatic, global down-regulation of the key TJ transmembrane protein occludin in regions of actively transmigrating PMN and in quiescent areas in the biopsy samples. Significant decreases in occludin expression were observed at the protein and mRNA levels by Western and Northern blotting. In contrast, expression of other TJ and AJ proteins such as ZO-1, claudin-1, JAM, beta-catenin, and E-cadherin were down-regulated only in epithelial cells immediately adjacent to transmigrating PMN. Analysis of inflamed mucosa from Crohn's disease patients mirrored the results obtained with UC patients. No change in TJ and AJ protein expression was observed in colonic epithelium from patients with collagenous colitis or lymphocytic colitis that are respectively characterized by a thickened subepithelial collagen plate and increased intraepithelial lymphocytes. These results suggest that occludin expression is diminished in IBD by mechanisms distinct from those regulating expression of other intercellular junction proteins. We speculate that down-regulation of epithelial occludin may play a role in enhanced paracellular permeability and PMN transmigration that is observed in active inflammatory bowel disease. PMID- 11733351 TI - Granulocytic sarcoma in MLL-positive infant acute myelogenous leukemia: fluorescence in situ hybridization study of childhood acute myelogenous leukemia for detecting MLL rearrangement. AB - Granulocytic sarcoma is considered to be rare and its frequent occurrence is associated with specific genetic changes such as t(8;21). To investigate an association between MLL (mixed lineage leukemia or myeloid-lymphoid leukemia) rearrangement and granulocytic sarcoma, we applied fluorescence in situ hybridization for detection of the 11q23/MLL rearrangements on the bone marrow cells of 40 patients with childhood acute myelogenous leukemia (AML). Nine (22.5%) of 40 patients exhibited MLL rearrangements. Three (33.3%) of these nine patients had granulocytic sarcoma and were younger than 12 months of age. Of these three patients one presented as granulocytic sarcoma of both testes with cerebrospinal fluid involvement, the second case presented in the form of an abdominal mass, and the third as a periorbital granulocytic sarcoma. On the other hand, no granulocytic sarcomas were found among MLL-negative patients. It is likely that MLL-positive infant AML may predispose granulocytic sarcoma. Regarding the findings of our study and those of other reports, we would guess that the incidence of granulocytic sarcoma in pediatric MLL-positive AML may be equal to or greater than the 18 to 24% described in AML with t(8;21). Further investigations designed to identify 11q23/MLL abnormalities of leukemic cells or extramedullary tumor may be helpful for the precise diagnosis of granulocytic sarcoma. PMID- 11733352 TI - STK11/LKB1 Peutz-Jeghers gene inactivation in intraductal papillary-mucinous neoplasms of the pancreas. AB - Despite the growing awareness of intraductal papillary-mucinous neoplasms (IPMNs) of the pancreas among clinicians, the molecular features of IPMNs have not been well characterized. Previous reports suggest that inactivation of the STK11/LKB1, a tumor-suppressor gene responsible for Peutz-Jeghers syndrome (PJS), plays a role in the pathogenesis of gastrointestinal hamartomas as well as several cancers, including pancreatic adenocarcinoma. Using polymerase chain reaction amplification of five microsatellite markers from the 19p13.3 region harboring the STK11/LKB1 gene, we analyzed DNA from 22 IPMNs for loss of heterozygosity (LOH). LOH at 19p13.3 was identified in 2 of 2 (100%) IPMNs from patients with PJS and 5 of 20 (25%) from patients lacking features of PJS (7 of 22, 32% overall). Sequencing analysis of the STK11/LKB1 gene in these IPMNs with LOH revealed a germline mutation in one IPMN that arose in a patient with PJS and a somatic mutation in 1 of the 20 sporadic IPMNs. None of the 22 IPMNs showed hypermethylation of the STK11/LKB1 gene. These results suggest that the STK11/LKB1 gene is involved in the pathogenesis of some IPMNs. PMID- 11733353 TI - Demonstration of light chain restricted clonal B-lymphoid infiltrates in archival bone marrow trephines by quantitative real-time polymerase chain reaction. AB - Assessment of clonality either by demonstrating light chain restriction or showing monoclonal immunoglobulin gene rearrangement is a valuable and indispensable adjunct to diagnosis in hematopathology. The study of light chain restriction by immunohistochemistry on archival material is hampered by a very low sensitivity especially regarding low grade lymphomas of B cell origin. DNA rearrangement studies of the immunoglobulin locus do improve sensitivity markedly but for lymphomas of follicle center origin they are prone to false negative results due to hypermutations. Therefore we developed a new clonality assay based on the quantification of immunoglobulin light chain transcripts using real-time polymerase chain reaction technology, which is also suitable for the analysis of archival bone marrow trephines. We tested the reproducibility and sensitivity of this approach by comparatively analyzing a series of bone marrow trephines with multiple myeloma (n = 26), reactive lymphoid hyperplasia (n = 37), and focal infiltration by low grade B cell lymphoma (n = 29). We could raise the detection rate of clonality from an average of 17% by immunohistochemistry and 66% as assessed by polymerase chain reaction rearrangement studies to 83% by this new technique. Despite false negative results due to light chain hypermutation in some cases, the detection rate of clonality could be improved even for B cell lymphomas of follicle center origin (follicular lymphoma or marginal zone lymphoma) thus making this novel approach a valuable additional tool for the hematopathologist. PMID- 11733354 TI - Simultaneous evaluation of T- and B-cell clonality, t(11;14) and t(14;18), in a single reaction by a four-color multiplex polymerase chain reaction assay and automated high-resolution fragment analysis: a method for the rapid molecular diagnosis of lymphoproliferative disorders applicable to fresh frozen and formalin-fixed, paraffin-embedded tissues, blood, and bone marrow aspirates. AB - Current polymerase chain reaction (PCR) methods for the molecular diagnosis of B- and T-cell lymphomas by determination of clonality of immunoglobulin heavy chain (IgH) and T-cell receptor-gamma rearrangements and by detection of the chromosomal translocations t(14;18) and t(11;14), require several laborious and costly PCR assays for each of these diagnostic tests. We have developed a multiplex PCR assay for the simultaneous determination of B- and T-cell clonality and the detection of the chromosomal translocations t(14;18) and t(11;14) in a single reaction, using four-color fluorescence and automated high-resolution fragment analysis. The 26 primers combined in the multiplex PCR correspond to the sequences of >90% of the 69 variables and 6 join IgH genes and 100% of the T-cell receptor-gamma variables and join genes that could participate in the respective rearrangements. In addition, they detect the major and the minor breakpoint regions of the t(14;18) and the major breakpoint region of the t(11;14), and amplify the beta-globin gene as an internal control. The specificity of the multiplex PCR was confirmed by analysis of 39 T-cell lymphomas and 58 B-cell lymphomas, including 11 mantle cell lymphomas bearing the t(11;14) and 25 follicular lymphomas bearing the t(14;18), with known rearrangements and/or translocations. Fifteen samples of reactive lymphadenitis remained negative. PMID- 11733355 TI - Cathepsin B knockout mice are resistant to tumor necrosis factor-alpha-mediated hepatocyte apoptosis and liver injury: implications for therapeutic applications. AB - Tumor necrosis factor-alpha (TNF-alpha) contributes to liver injury by inducing hepatocyte apoptosis. Recent evidence suggests that cathepsin B (cat B) contributes to TNF-alpha-induced apoptosis in vitro. The aim of the present study was to determine whether cat B contributes to TNF-alpha-induced hepatocyte apoptosis and liver injury in vivo. Cat B knockout (catB(-/-)) and wild-type (catB(+/+)) mice were first infected with the adenovirus Ad5I kappa B expressing the I kappa B superrepressor to inhibit nuclear factor-kappa B-induced survival signals and then treated with murine recombinant TNF-alpha. Massive hepatocyte apoptosis with mitochondrial release of cytochrome c and activation of caspases 9 and 3 was detected in catB(+/+) mice 2 hours after the injection of TNF-alpha. In contrast, significantly less hepatocyte apoptosis and no detectable release of cytochrome c or caspase activation occurred in the livers of catB(-/-) mice. By 4 hours after TNF-alpha injection, only 20% of the catB(+/+) mice were alive as compared to 85% of catB(-/-) mice. Pharmacological inhibition of cat B in catB(+/+) mice with L-3-trans-(propylcarbamoyl)oxirane-2-carbonyl-L-isoleucyl-L proline (CA-074 Me) also reduced TNF-alpha-induced liver damage. The present data demonstrate that a cat B-mitochondrial apoptotic pathway plays a pivotal role in TNF-alpha-induced hepatocyte apoptosis and liver injury. PMID- 11733356 TI - Effects of paramyxoviral infection on airway epithelial cell Foxj1 expression, ciliogenesis, and mucociliary function. AB - To elucidate molecular mechanisms underlying the association between respiratory viral infection and predisposition to subsequent bacterial infection, we used in vivo and in vitro models and human samples to characterize respiratory virus induced changes in airway epithelial cell morphology, gene expression, and mucociliary function. Mouse paramyxoviral bronchitis resulted in airway epithelial cell infection and a distinct pattern of epithelial cell morphology changes and altered expression of the differentiation markers beta-tubulin-IV, Clara cell secretory protein, and Foxj1. Furthermore, changes in gene expression were recapitulated using an in vitro epithelial cell culture system and progressed independent of the host inflammatory response. Restoration of mature airway epithelium occurred in a pattern similar to epithelial cell differentiation and ciliogenesis in embryonic lung development characterized by sequential proliferation of undifferentiated cells, basal body production, Foxj1 expression, and beta-tubulin-IV expression. The effects of virus-induced alterations in morphology and gene expression on epithelial cell function were illustrated by decreased airway mucociliary velocity and impaired bacterial clearance. Similar changes in epithelial cell Foxj1 expression were also observed in human paramyxoviral respiratory infection. Taken together, these model systems of paramyxoviral respiratory infection mimic human pathology and identify epithelial cell Foxj1 expression as an early marker of epithelial cell differentiation, recovery, and function. PMID- 11733357 TI - Beta 1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids. AB - Ovarian carcinoma multicellular spheroids are an in vitro model of micrometastasis whose adhesive abilities have not been elucidated. In this study, we identified adhesion molecules that mediate the formation of ovarian carcinoma spheroids and their subsequent adhesion to extracellular matrix proteins. The NIH:OVCAR5, but not the SKOV3, ovarian carcinoma cell line formed spheroids similar to multicellular aggregates isolated from patient ascitic fluid. NIH:OVCAR5 spheroid formation was augmented by a beta 1-integrin-stimulating monoclonal antibody or exogenous fibronectin, but was inhibited by blocking monoclonal antibodies against the alpha 5- or beta 1-integrin subunits. By immunohistochemical staining, alpha 2-, alpha 3-, alpha 5-, alpha 6-, and beta 1 integrin subunits, CD44, and fibronectin were detected in NIH:OVCAR5 spheroids. NIH:OVCAR5 spheroids adhered to fibronectin, laminin, and type IV collagen, and this adhesion was partially inhibited by blocking antibodies against the alpha 5 , alpha 6-, and alpha 2-integrin subunits, respectively. A blocking monoclonal antibody against the beta 1-integrin subunit completely inhibited adhesion of the spheroids to all three proteins. These results suggest that interactions between the alpha 5 beta 1-integrin and fibronectin mediate the formation of ovarian carcinoma spheroids and that their adhesion to extracellular matrix proteins at sites of secondary tumor growth may be mediated by a complex interaction between multiple integrins and their ligands. PMID- 11733359 TI - Amplification of EIF3S3 gene is associated with advanced stage in prostate cancer. AB - Gain of the long arm of chromosome 8 (8q) is one of the most common gains found in the advanced prostate cancer by comparative genomic hybridization. We have previously identified a putative target gene for the 8q gain, EIF3S3, that encodes a p40 subunit of eukaryotic translation initiation factor 3 (eIF3). Here, we studied the frequency of the EIF3S3 amplification in different stages of prostate cancer and co-amplification of EIF3S3 and oncogene MYC. In addition, prognostic utility of the EIF3S3 copy number alteration was evaluated. The analyses were done with fluorescence in situ hybridization and tissue microarray technology. High-level amplification of EIF3S3 was found in 11 of 125 (9%) of pT1/pT2 tumors, 12 of 44 (27%) of pT3/pT4 tumors, and 8 of 37 (22%) of lymph node metastases as well as in 26 of 78 (33%) and 15 of 30 (50%) of hormone refractory locally recurrent tumors and metastases, respectively. The amplification was associated with high Gleason score (P < 0.001). One of the 79 tumors with EIF3S3 amplification had only two copies of MYC, whereas all tumors with amplification of MYC had also amplification of EIF3S3 indicating common co-amplification of the genes. Gain of EIF3S3 was associated with poor cancer-specific survival in incidentally found prostate carcinomas (P = 0.023). In the analyses of prostatectomy-treated patients, the amplification was not statistically significantly associated with progression-free time. In conclusion, amplification of EIF3S3 gene is common in late-stage prostate cancer suggesting that it may be functionally involved in the progression of the disease. PMID- 11733358 TI - Immunohistochemical detection of myeloperoxidase and its oxidation products in Kupffer cells of human liver. AB - Oxidative damage to tissue proteins has been implicated in the pathogenesis of liver disease, but the mechanisms that promote oxidation in vivo are unclear. Hydrogen peroxide is transformed into an array of potentially damaging reactants by the heme protein myeloperoxidase. This proinflammatory enzyme is expressed by circulating neutrophils and monocytes but is generally thought to be absent from tissue macrophages. To determine whether myeloperoxidase is present in Kupffer cells, the fixed-tissue macrophages of liver, Western blot analysis, and immunohistochemistry were performed. Two different antibodies monospecific for myeloperoxidase identified a 60-kd protein, the predicted molecular mass of myeloperoxidase, in human liver extracts. Immunostaining detected the enzyme in sinusoidal lining cells of normal and diseased human livers. Immunofluorescence confocal microscopy demonstrated co-localization of myeloperoxidase and CD68, a monocyte/macrophage marker, in sinusoidal lining cells. Numerous myeloperoxidase expressing cells were also evident in the fibrous septa of cirrhotic livers. Immunostaining with an antibody to proteins modified by hypochlorous acid, a characteristic product of the enzyme, indicated that myeloperoxidase is enzymatically active in cases of acute liver injury and cirrhosis. These findings identify myeloperoxidase as a component of human Kupffer cells. Oxidative damage resulting from the action of myeloperoxidase may contribute to acute liver injury and hepatic fibrogenesis. PMID- 11733360 TI - p53 Mutations in nasal natural killer/T-cell lymphoma from Mexico: association with large cell morphology and advanced disease. AB - Nasal NK/T-cell lymphoma is a unique form of lymphoma highly associated with Epstein-Barr virus, and with a characteristic geographic distribution. Recently, we showed that p53 is overexpressed in a high percentage of nasal NK/T-cell lymphomas. The aim of this study was to analyze the status of the p53 gene, and correlate it with the expression of p53 protein and its downstream target, the cyclin-dependent kinase inhibitor p21, in a series of 25 cases of well characterized nasal NK/T-cell lymphoma from Mexico. The highly conserved exons 5 to 8 of the p53 gene were amplified by polymerase chain reaction and screened for mutations by denaturing high-pressure liquid chromatography. Abnormal polymerase chain reaction products detected by denaturing high-pressure liquid chromatography and additional selected cases were sequenced. In addition, the incidence of loss of heterozygosity at the p53 locus was analyzed in 12 cases. Of the 25 patients, 17 were male and 8 female (M:F ratio, 2.1:1), with a median age of 43 years (range, 21 to 93 years). Morphologically, most of the cases were composed of a mixture of medium-sized cells and large transformed cells (21 cases), and four cases were composed exclusively of large transformed cells. Three different groups determined by p53 gene status and expression of p53 protein were identified: group 1 was p53 +/p53 mutated (five cases, all with p53 missense mutations). Morphologically, three of the five cases were composed of large cells. All five cases revealed overexpression of p53 in the majority of the tumor cells with a mean of 86%. Unexpectedly, three of these cases also showed overexpression of p21. Four of the five patients presented with clinical stage IVB and died with disease. Group 2 was p53+/p53 wild-type (10 cases). Histologically, nine cases were of the mixed type, and one of the large cell type. The percentage of p53 overexpressing cells was lower than in the previous group with a mean of 23%. p21 was positive in 7 of the 10 cases. Six patients in this group presented with clinical stages I to II and four patients with advanced disease (stage III and IV). Five patients are alive 12 to 120 months later (mean, 24 months), three with no evidence of disease. Group 3 was p53-/p53 wild-type (10 cases). All cases showed mixed cell morphology. p21 was positive in 5 of 10 cases. Four patients presented with clinical stage I to II and six patients with advanced disease. Four patients are alive with no evidence of disease 9 to 60 months later (mean, 10 months). Overall, p53 mutations were present in 24% (5 of 21) of the evaluable cases, all of them overexpressing p53 in the majority of tumor cells. Cases with p53 mutations were associated with large cell morphology (P = 0.0162) and presented more often with advanced stage disease. Loss of heterozygosity at chromosome 17p was found only in 2 of the 12 (17%) cases investigated, both cases showed p53 mutations of the remaining allele. P21 overexpression (60% of cases) is frequent in nasal NK/T-cell lymphoma and seems to be independent of p53 gene status. The overexpression of p53 and p21, independent of p53 mutations, although as yet not clear, might be the result of Epstein-Barr virus infection, and warrants further investigation. PMID- 11733361 TI - Features of colorectal cancers with high-level microsatellite instability occurring in familial and sporadic settings: parallel pathways of tumorigenesis. AB - High-level microsatellite instability (MSI-H) is demonstrated in 10 to 15% of sporadic colorectal cancers and in most cancers presenting in the inherited condition hereditary nonpolyposis colorectal cancer (HNPCC). Distinction between these categories of MSI-H cancer is of clinical importance and the aim of this study was to assess clinical, pathological, and molecular features that might be discriminatory. One hundred and twelve MSI-H colorectal cancers from families fulfilling the Bethesda criteria were compared with 57 sporadic MSI-H colorectal cancers. HNPCC cancers presented at a lower age (P < 0.001) with no sporadic MSI H cancer being diagnosed before the age of 57 years. MSI was less extensive in HNPCC cancers with 72% microsatellite markers showing band shifts compared with 87% in sporadic tumors (P < 0.001). Absent immunostaining for hMSH2 was only found in HNPCC tumors. Methylation of hMLH1 was observed in 87% of sporadic cancers but also in 55% of HNPCC tumors that showed loss of expression of hMLH1 (P = 0.02). HNPCC cancers were more frequently characterized by aberrant beta catenin immunostaining as evidenced by nuclear positivity (P < 0.001). Aberrant p53 immunostaining was infrequent in both groups. There were no differences with respect to 5q loss of heterozygosity or codon 12 K-ras mutation, which were infrequent in both groups. Sporadic MSI-H cancers were more frequently heterogeneous (P < 0.001), poorly differentiated (P = 0.02), mucinous (P = 0.02), and proximally located (P = 0.04) than HNPCC tumors. In sporadic MSI-H cancers, contiguous adenomas were likely to be serrated whereas traditional adenomas were dominant in HNPCC. Lymphocytic infiltration was more pronounced in HNPCC but the results did not reach statistical significance. Overall, HNPCC cancers were more like common colorectal cancer in terms of morphology and expression of beta catenin whereas sporadic MSI-H cancers displayed features consistent with a different morphogenesis. No individual feature was discriminatory for all HNPCC cancers. However, a model based on four features was able to classify 94.5% of tumors as sporadic or HNPCC. The finding of multiple differences between sporadic and familial MSI-H colorectal cancer with respect to both genotype and phenotype is consistent with tumorigenesis through parallel evolutionary pathways and emphasizes the importance of studying the two groups separately. PMID- 11733362 TI - E-cadherin gene mutations frequently occur in synovial sarcoma as a determinant of histological features. AB - Synovial sarcoma is a mesenchymal tumor that has an epithelial character and two major histological subtypes, the biphasic type and the monophasic fibrous type. However, the mechanisms involved in its epithelial differentiation are unknown, and furthermore, the determinants for histological subtype in synovial sarcoma remain unclear. In this study, we immunohistochemically examined E-cadherin expression and screened for genetic alterations in the E-cadherin gene from exon 4 to exon 9 in 49 cases of synovial sarcoma. In addition, we also examined the mRNA expressions of E-cadherin and Snail, a direct repressor of E-cadherin gene expression, by reverse transcriptase-polymerase chain reaction in 20 samples of frozen material. Immunohistochemical E-cadherin membranous expression was observed in 12 cases (24.5%), and was predominant in biphasic tumors. Single strand conformation polymorphism analysis followed by DNA direct sequencing revealed 15 missense E-cadherin mutations in 12 cases (24.5%: monophasic, 11 of 42; biphasic, 1 of 6; poorly, 0 of 1) and 7 silent mutations (14.3%) in 7 cases. Ten of the 12 cases with E-cadherin missense mutations did not show E-cadherin membranous expression. Reverse transcriptase-polymerase chain reaction demonstrated E-cadherin and Snail mRNA expressions in 14 cases (70%) and in all cases, respectively. E-cadherin gene expression was inactivated by missense mutations in three of the eight cases (37.5%) of monophasic fibrous tumors that showed E-cadherin mRNA expressions. The E-cadherin gene was potentially inactivated in a significant number of synovial sarcomas. E-cadherin dysfunction because of its mutation in the central region of the molecule was associated with its decreased immunohistochemical expression and histological fibroblastic and spindle-shaped features of monophasic tumors. Thus, E-cadherin gene mutation may be one of the determinants of histological subtype in synovial sarcoma. PMID- 11733363 TI - Expression of matrix metalloproteinases in invasive pulmonary adenocarcinoma with bronchioloalveolar component and atypical adenomatous hyperplasia. AB - To investigate the association between the expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) and the clinicopathological features in lepidic and invasive components of adenocarcinoma of the lung, we performed immunostaining for type IV collagen, various MMPs, and TIMPs in 27 cases of invasive adenocarcinomas and 5 cases of atypical adenomatous hyperplasia of alveolar epithelial cells (AAH). Mean extent of lepidic growth was 61% and the survival was significantly better in cases with 50% or more lepidic component. The preservation of type IV collagen in lepidic areas correlated inversely with lymphatic or vascular invasion (P = 0.02 and 0.002, respectively). Five-year survival was reduced in cases showing destruction of type IV collagen (P = 0.004) or expression of MMP-2 (P = 0.008) in lepidic areas. MMP-2 co localized with MT-1-MMP (its activating enzyme) and TIMP-2 in neoplastic cells. Reactivity for other MMPs and TIMPs did not correlate with destruction of type IV collagen or prognosis. Type IV collagen was preserved in all cases of AAH. MMP-2, but not MT-1-MMP, was expressed in two of the five cases of AAH. Immunostaining for type IV collagen MMP-2 is useful in evaluating the prognosis of the lung. PMID- 11733364 TI - Neutralization of Gro alpha and macrophage inflammatory protein-2 attenuates renal ischemia/reperfusion injury. AB - Previous studies have provided strong evidence for a role for neutrophils in mediating pathology during reperfusion of ischemic tissues. CXC chemokines including interleukin-8, KC/Gro alpha, and macrophage inflammatory protein (MIP) 2, direct neutrophils to tissue sites of inflammation. In the current study we tested the efficacy of antibodies to KC/Gro alpha and MIP-2 in inhibiting neutrophil infiltration into kidneys during reperfusion after 1 hour of warm ischemia using a mouse model. KC mRNA and protein were produced within 3 hours after reperfusion of the ischemic kidneys. MIP-2 mRNA and protein were twofold to fourfold lower than KC and were at low levels until 9 hours after reperfusion. Only 60% of mice subjected to ischemia/reperfusion injury survived to day 3 after reperfusion. Treatment with rabbit neutralizing antibodies to both KC and MIP-2 inhibited neutrophil infiltration into ischemic kidneys during reperfusion, restored renal function as assessed by decreased serum creatinine and urea nitrogen levels to near normal levels, and resulted in complete survival of treated animals. Finally, treatment with both antibodies significantly reduced histologically graded pathology of kidneys subjected to ischemia/reperfusion injury. Collectively, the results indicate the efficacy of neutralizing the chemokines directing neutrophils into ischemic kidneys during reperfusion to inhibit this infiltration and attenuate the resulting pathology. PMID- 11733366 TI - Interleukin-6 is an autocrine growth factor in human prostate cancer. AB - Prostate cancer is the most common cancer in American men and the second leading cause of cancer deaths in this group. We have found that interleukin (IL)-6 protein concentrations are increased approximately 18-fold in clinically localized prostate cancers when compared to normal prostate tissue. Normal and neoplastic prostatic epithelial cells in culture, with the exception of LNCaP cells, secrete IL-6. Addition of exogenous IL-6 to primary epithelial cells in culture or the LNCaP prostate cancer cell line leads to phosphorylation of Stat-3 and increases in net cell proliferation. The concentration of IL-6 receptor is increased eightfold in the prostate cancer tissues and is increased in the cancer cells by immunohistochemistry. The increased expression of IL-6 receptor is correlated with increased proliferation of prostate cancer cells in vivo as assessed by Ki67 immunohistochemistry. These findings strongly support the hypothesis that IL-6 acts as a significant autocrine growth factor in vivo for primary, androgen-dependent prostate cancers. PMID- 11733365 TI - Healing of burn wounds in transgenic mice overexpressing transforming growth factor-beta 1 in the epidermis. AB - Transforming growth factor-beta (TGF-beta) isoforms are multifunctional cytokines that play an important role in wound healing. Transgenic mice overexpressing TGF beta in the skin under control of epidermal-specific promoters have provided models to study the effects of increased TGF-beta on epidermal cell growth and cutaneous wound repair. To date, most of these studies used transgenic mice that overexpress active TGF-beta in the skin by modulating the latency-associated peptide to prevent its association with active TGF-beta. The present study is the first to use transgenic mice that overexpress the natural form of latent TGF-beta 1 in the epidermis, driven by the keratin 14 gene promoter to investigate the effects of locally elevated TGF-beta 1 on the healing of partial-thickness burn wounds made on the back of the mice using a CO(2) laser. Using this model, we demonstrated activation of latent TGF-beta after wounding and determined the phenotypes of burn wound healing. We found that introduction of the latent TGF beta1 gene into keratinocytes markedly increases the release and activation of TGF-beta after burn injury. Elevated local TGF-beta significantly inhibited wound re-epithelialization in heterozygous (42% closed versus 92% in controls, P < 0.05) and homozygous (25% versus 92%, P < 0.01) animals at day 12 after wounding. Interestingly, expression of type I collagen mRNA and hydroxyproline significantly increased in the wounds of transgenic mice, probably as a result of a paracrine effect of the transgene. PMID- 11733367 TI - Cathepsin k is a critical protease in synovial fibroblast-mediated collagen degradation. AB - Synovial fibroblasts (SFs) play a critical role in the pathogenesis of rheumatoid arthritis (RA) and are directly involved in joint destruction. Both SF-resident matrix metalloproteases and cathepsins have been implicated in cartilage degradation although their identities and individual contributions remain unclear. The aims of this study were to investigate the expression of cathepsin K in SFs, the correlation between cathepsin K expression and disease severity, and the contribution of cathepsin K to fibroblast-mediated collagen degradation. Immunostaining of joint specimens of 21 patients revealed high expression of cathepsin K in SFs in the synovial lining and the stroma of synovial villi, and to a lesser extent in CD68-positive cells of the synovial lining. Cathepsin K positive SFs were consistently observed at sites of cartilage and bone degradation. Expression levels of cathepsin K in the sublining and vascularized areas of inflamed synovia showed a highly significant negative correlation with results derived from the Hannover Functional Capacity Questionnaire (r = 0.78, P = 0.003; and r = 0.70, P = 0.012, respectively) as a measure of the severity of RA in individual patients. For comparison, there was no correlation between Hannover Functional Capacity Questionnaire and cathepsin S whose expression is limited to CD-68-positive macrophage-like synoviocytes. The expression of cathepsin K was also demonstrated in primary cell cultures of RA-SFs. Co-cultures of SFs on cartilage disks revealed the ability of fibroblast-like cells to phagocytose collagen fibrils whose intralysosomal hydrolysis was prevented in the presence of a potent cathepsin K inhibitor but not by an inhibitor effective against cathepsins L, B, and S. The selective and critical role of cathepsin K in articular cartilage and subchondral bone erosion was further corroborated by the finding that cathepsin K has a potent aggrecan-degrading activity and that cathepsin K-generated aggrecan cleavage products specifically potentiate the collagenolytic activity of cathepsin K toward type I and II collagens. This study demonstrates for the first time a critical role of cathepsin K in cartilage degradation by SFs in RA that is comparable to its well-known activity in osteoclasts. PMID- 11733368 TI - Plasminogen deficiency leads to impaired lobular reorganization and matrix accumulation after chronic liver injury. AB - To determine the regulatory role of plasminogen in hepatic repair following a chronic liver injury, we injected carbon tetrachloride (CCl(4)) biweekly into mice lacking plasminogen (Plg(0)) and plasminogen-sufficient littermates (Plg(+)). On gross examination, we found that Plg(0) livers became enlarged and pale with foci of red nodules as early as 4 weeks after CCl(4) injection, while Plg(+) livers appeared minimally affected by 6 weeks. Microscopically, Plg(0) livers had a pronounced pericentral linking, with accumulation of centrilobular eosinophilic material in injured areas, which resulted in a significant increase in liver mass and total protein. Immunohistochemistry revealed that fibrin accumulated progressively in injured regions. However, the combined genetic loss of plasminogen and fibrinogen did not correct the abnormal phenotype. Mason's trichrome staining revealed intense signal in centrilobular regions and electron microscopy showed a marked increase in fibrillary material demonstrating an excessive accumulation of extracellular matrix in Plg(0) mice. The zone-specific increase in matrix components was associated with an increase in the number of activated hepatic stellate cells within injured sites of Plg(0) livers. Taken together, these data suggest that the progressive accumulation of fibrin unrelated matrix substrates in Plg(0) livers after a chronic injury results from the combined effects of impaired proteolysis and increased matrix production. PMID- 11733369 TI - Rapid response of identified resident endoneurial macrophages to nerve injury. AB - Macrophages play a central role in the pathogenesis of peripheral neuropathy but the role of resident endoneurial macrophages is undefined because no discriminating markers exist to distinguish them from infiltrating hematogenous macrophages. We identified and characterized resident endoneurial macrophages during Wallerian degeneration in radiation bone marrow chimeric rats created by transplanting wild-type Lewis rat bone marrow into irradiated TK-tsa transgenic Lewis rats. In such animals, resident cells carry the transgene, whereas hematogenous cells do not. As early as 2 days after sciatic nerve crush and before the influx of hematogenous macrophages, resident transgene-positive endoneurial macrophages underwent morphological and immunophenotypic signs of activation. At the same time, resident macrophages phagocytosing myelin were found, and proliferation was detected by bromodeoxyuridine incorporation. Continuous bromodeoxyuridine feeding revealed that resident endoneurial macrophages sequentially retracted their processes, proliferated, and expressed the ED1 antigen, rendering them morphologically indistinguishable from hematogenous macrophages. Resident endoneurial macrophages thus play an early and active role in the cellular events after nerve lesion before hematogenous macrophages enter the nerve. They may thus be critically involved in the pathogenesis of peripheral neuropathy particularly at early stages of the disease and may act as sensors of pathology much like their central nervous system counterparts, the microglial cells. PMID- 11733370 TI - Chemokine ligand and receptor expression in the pregnant uterus: reciprocal patterns in complementary cell subsets suggest functional roles. AB - During human pregnancy, the uterus is infiltrated by a population of maternal leukocytes that co-exist with fetal cytotrophoblasts occupying the decidua and uterine blood vessels. These immune cells, termed "decidual granulated leukocytes," are composed predominantly (70%) of the CD56(bright) subset of natural killer cells, accompanied by T cells (15%) and macrophages (15%). The mechanisms underlying the recruitment of these cells are unknown, but by analogy to other systems, chemokines are likely to be involved. We examined the expression patterns of 14 chemokines in the decidualized uterine wall by in situ hybridization, and the expression of chemokine receptors on decidual leukocytes by RNase protection. The striking concordance between the expression of chemokines in the uterus and their receptors on decidual leukocytes allowed us to identify numerous receptor-ligand pairs that may recruit the latter cells to the uterus during pregnancy. Additionally, chemokine expression patterns suggested other, nonimmune functions for these molecules, including a role in cytotrophoblast differentiation. Together, our results imply that chemokine networks serve important functions at the maternal-fetal interface. PMID- 11733371 TI - Selective insolubility of alpha-synuclein in human Lewy body diseases is recapitulated in a transgenic mouse model. AB - alpha-Synuclein (alpha-SYN) is deposited in intraneuronal cytoplasmic inclusions (Lewy bodies, LBs) characteristic for Parkinson's disease (PD) and LB dementias. alpha-SYN forms LB-like fibrils in vitro, in contrast to its homologue beta-SYN. Here we have investigated the solubility of SYNs in human LB diseases and in transgenic mice expressing human wild-type and PD-associated mutant [A30P]alpha SYN driven by the brain neuron-specific promoter, Thy1. Distinct alpha-SYN species were detected in the detergent-insoluble fractions from brains of patients with PD, dementia with LBs, and neurodegeneration with brain iron accumulation type 1 (formerly known as Hallervorden-Spatz disease). Using the same extraction method, detergent-insolubility of human alpha-SYN was observed in brains of transgenic mice. In contrast, neither endogenous mouse alpha-SYN nor beta-SYN were detected in detergent-insoluble fractions from transgenic mouse brains. The nonamyloidogenic beta-SYN was incapable of forming insoluble fibrils because amino acids 73 to 83 in the central region of alpha-SYN are absent in beta-SYN. In conclusion, the specific accumulation of detergent-insoluble alpha SYN in transgenic mice recapitulates a pivotal feature of human LB diseases. PMID- 11733372 TI - Coordinated induction of extracellular proteolysis systems during experimental autoimmune encephalomyelitis in mice. AB - Plasminogen activators (PAs) and matrix metalloproteinases (MMPs) are considered to play an important role in the pathogenesis of multiple sclerosis. Experimental autoimmune encephalomyelitis (EAE) is widely used as an animal model of multiple sclerosis. Whereas several studies have addressed the expression of various MMPs and their inhibitors in the pathogenesis of EAE, the expression of the molecules of the PA system during EAE has not been reported previously. The present study was undertaken to investigate the expression of the molecules of the PA system (tPA, uPA, PAI-1, uPAR, LRP), as well as several members of the MMP family and their inhibitors in the course of actively induced EAE in BALB/c mice. During clinical EAE, the PA system was up-regulated in the central nervous system at several levels. Induction of expression of tPA and PAI-1 transcripts was detected in activated astrocytes in the white matter. Inflammatory cells expressed uPA receptor, uPAR. In situ zymography demonstrated the presence of increased tPA and uPA activities in the areas of the inflammatory damage. Accumulation of fibrin, fibronectin, and vitronectin immunoreactivity was seen in perivascular matrices of symptomatic animals. In addition, transcription of MT1-MMP and metalloelastase (in inflammatory cells), and TIMP-1 (in activated astrocytes) was induced during EAE. Increased gelatinolytic activity was detected at the sites of inflammatory cell accumulation by in situ zymography of fluorescently labeled gelatin; substrate gel zymography identified the up-regulated gelatinolytic activity as gelatinase B. Overall, our study demonstrates concurrent induction of PA and MMP systems during active EAE, supporting further the concept that the neuroinflammatory damage in EAE involves altered balance between multiple extracellular proteases and their inhibitors. PMID- 11733374 TI - Tissue microarrays for rapid linking of molecular changes to clinical endpoints. AB - Advances in genomics and proteomics are dramatically increasing the need to evaluate large numbers of molecular targets for their diagnostic, predictive or prognostic value in clinical oncology. Conventional molecular pathology techniques are often tedious, time-consuming, and require a lot of tissue, thereby limiting both the number of tissues and the number of targets that can be evaluated. Here, we demonstrate the power of our recently described tissue microarray (TMA) technology in analyzing prognostic markers in a series of 553 breast carcinomas. Four independent TMAs were constructed by acquiring 0.6 mm biopsies from one central and from three peripheral regions of each of the formalin-fixed paraffin embedded tumors. Immunostaining of TMA sections and conventional "large" sections were performed for two well- established prognostic markers, estrogen receptor (ER) and progesterone receptor (PR), as well as for p53, another frequently examined protein for which the data on prognostic utility in breast cancer are less unequivocal. Compared with conventional large section analysis, a single sample from each tumor identified about 95% of the information for ER, 75 to 81% for PR, and 70 to 74% for p53. However, all 12 TMA analyses (three antibodies on four different arrays) yielded as significant or more significant associations with tumor-specific survival than large section analyses (p < 0.0015 for each of the 12 comparisons). A single sample from each tumor was sufficient to identify associations between molecular alterations and clinical outcome. It is concluded that, contrary to expectations, tissue heterogeneity did not negatively influence the predictive power of the TMA results. TMA technology will be of substantial value in rapidly translating genomic and proteomics information to clinical applications. PMID- 11733373 TI - Loss of CDX2 expression and microsatellite instability are prominent features of large cell minimally differentiated carcinomas of the colon. AB - Most large bowel cancers are moderately to well-differentiated adenocarcinomas comprised chiefly or entirely of glands lined by tall columnar cells. We have identified a subset of poorly differentiated colon carcinomas with a distinctive histopathological appearance that we term large cell minimally differentiated carcinomas (LCMDCs). These tumors likely include a group of poorly differentiated carcinomas previously described by others as medullary adenocarcinomas. To better understand the pathogenesis of these uncommon neoplasms, we compared molecular features of 15 LCMDCs to those present in 25 differentiated adenocarcinomas (DACs) of the colon. Tumors were examined for alterations commonly seen in typical colorectal carcinomas, including increased p53 and beta-catenin immunoreactivity, K-ras gene mutations, microsatellite instability, and loss of heterozygosity of markers on chromosomes 5q, 17p, and 18q. In addition, tumors were evaluated by immunohistochemistry for CDX2, a homeobox protein whose expression in normal adult tissues is restricted to intestinal and colonic epithelium. Markedly reduced or absent CDX2 expression was noted in 13 of 15 (87%) LCMDCs, whereas only 1 of the 25 (4%) DACs showed reduced CDX2 expression (P < 0.001). Nine of 15 (60%) LCMDCs had the high-frequency microsatellite instability phenotype, but only 2 of 25 (8%) DACs had the high-frequency microsatellite instability phenotype (P = 0.002). Our findings provide support for the hypothesis that the molecular pathogenesis of LCMDCs is distinct from that of most DACs. CDX2 alterations and DNA mismatch repair defects have particularly prominent roles in the development of LCMDCs. PMID- 11733375 TI - A dominant interference collagen X mutation disrupts hypertrophic chondrocyte pericellular matrix and glycosaminoglycan and proteoglycan distribution in transgenic mice. AB - Collagen X transgenic (Tg) mice displayed skeleto-hematopoietic defects in tissues derived by endochondral skeletogenesis.(1) Here we demonstrate that co expression of the transgene product containing truncated chicken collagen X with full-length mouse collagen X in a cell-free translation system yielded chicken mouse hybrid trimers and truncated chicken homotrimers; this indicated that the mutant could assemble with endogenous collagen X and thus had potential for dominant interference. Moreover, species-specific collagen X antibodies co localized the transgene product with endogenous collagen X to hypertrophic cartilage in growth plates and ossification centers; proliferative chondrocytes also stained diffusely. Electron microscopy revealed a disrupted hexagonal lattice network in the hypertrophic chondrocyte pericellular matrix in Tg growth plates, as well as altered mineral deposition. Ruthenium hexamine trichloride positive aggregates, likely glycosaminoglycans (GAGs)/proteoglycans (PGs), were also dispersed throughout the chondro-osseous junction. These defects likely resulted from transgene co-localization and dominant interference with endogenous collagen X. Moreover, altered GAG/PG distribution in growth plates of both collagen X Tg and null mice was confirmed by a paucity of staining for hyaluronan and heparan sulfate PG. A provocative hypothesis links the disruption of the collagen X pericellular network and GAG/PG decompartmentalization to the potential locus for hematopoietic failure in the collagen X mice. PMID- 11733376 TI - Increased Tie2 expression, enhanced response to angiopoietin-1, and dysregulated angiopoietin-2 expression in hemangioma-derived endothelial cells. AB - Infantile hemangiomas are endothelial tumors that grow rapidly in the first year of life and regress slowly during early childhood. Although hemangiomas are well known vascular lesions, little is known about the mechanisms that cause the excessive endothelial cell proliferation in these most common tumors of infancy. To investigate the molecular basis of hemangioma, we isolated endothelial cells from several proliferative-phase lesions and showed that these cells are clonal and exhibit abnormal properties in vitro (E. Boye, Y. Yu, G. Paranya, J. B. Mulliken, B. R. Olsen, J. Bischoff: Clonality and altered behavior of endothelial cells from hemangiomas. J Clin Invest 2001, 107:745-752). Here, we analyzed mRNA expression patterns of genes required for angiogenesis, including members of the vascular endothelial growth factor (VEGF)/VEGF receptor family and the angiopoietin/Tie family, in hemangioma-derived and normal endothelial cells. KDR, Flt-1, Tie1, Tie2, and angiopoietin-2 (Ang2) were strongly expressed in cultured hemangioma-derived endothelial cells and in hemangioma tissue. In contrast, there was little expression of angiopoietin-1 (Ang1) or VEGF. We found Tie2 mRNA and protein up-regulated with a concomitant increase in cellular responsiveness to Ang1 in most hemangioma-derived endothelial cells. Ang2 mRNA was down-regulated in response to serum in hemangioma-derived endothelial cells, but not in normal endothelial cells, suggesting altered regulation. These findings implicate Tie2 and its ligands Ang1 and Ang2 in the pathogenesis of hemangioma. PMID- 11733377 TI - Reduced expression of junctional adhesion molecule and platelet/endothelial cell adhesion molecule-1 (CD31) at human vascular endothelial junctions by cytokines tumor necrosis factor-alpha plus interferon-gamma Does not reduce leukocyte transmigration under flow. AB - The combination of tumor necrosis factor (TNF)-alpha plus interferon (IFN)-gamma has been shown previously to promote redistribution of platelet/endothelial cell adhesion molecule-1 (PECAM-1) (CD31), junctional adhesion molecule (JAM), and VE cadherin away from lateral junctions of human umbilical vein endothelial cell monolayers. In parallel, neutrophil transmigration was significantly reduced. Because PECAM-1 and JAM have been implicated in leukocyte transmigration, the observed redistribution by cytokine activation was presumed to represent the mechanism causing decreased transmigration under static conditions. The current results confirm that culture of human umbilical vein endothelial cells with TNF alpha plus IFN-gamma caused a decrease in surface-expressed and junctional localized JAM and PECAM-1, but did not cause decreased leukocyte transmigration in an in vitro flow assay. Furthermore, blocking monoclonal antibody to PECAM-1 still significantly reduced monocyte transmigration, demonstrating that it retains a functional role even though its levels were reduced and redistributed away from junctions, whereas a panel of monoclonal antibodies to JAM failed to reduce leukocyte transmigration. Given the alterations in junction protein location, permeability function was assessed. IFN-gamma alone or TNF-alpha plus IFN-gamma significantly increased permeability, but TNF-alpha alone did not, suggesting lack of correlation between transmigration and loss of permeability. In conclusion, cytokine activation induced loss and redistribution of PECAM-1 and JAM away from lateral junctions, but per se does not negatively regulate either neutrophil or monocyte transmigration under flow. PMID- 11733378 TI - Identification of receptor-binding sites of monocyte chemotactic S19 ribosomal protein dimer. AB - The S19 ribosomal protein (RP S19) cross-linked homo-dimer attracts monocyte migration by binding to C5a receptor on monocytes (H Nishiura, Y Shibuya, T Yamamoto, Laboratory Investigation, 1998, 78:1615-1623). Using site-directed mutants of recombinant RP S19 and synthetic peptides mimicking RP S19 molecular regions, we currently identified the binding sites of the RP S19 dimer to the C5a receptor. The RP S19 dimer activated the receptor by a two-step binding mechanism as in the case of C5a. The first binding site was a basic cluster region containing a -Lys41-His42-Lys43- sequence. The second one was the -Leu131-Asp132 Arg133- moiety, localized 12 residues upstream from the COOH-terminal. The second binding triggered the chemotactic response. The first binding would have a role in achieving a high-binding affinity between the ligand and receptor. The first and second ligand-binding sites of C5a receptor seem to be shared by C5a and the RP S19 dimer, although overall homology between the amino acid sequences of these ligands is only 4%. PMID- 11733379 TI - CD2AP localizes to the slit diaphragm and binds to nephrin via a novel C-terminal domain. AB - CD2AP, an adapter protein containing multiple SH3 domains, plays a critical role in kidney function. Mice lacking CD2AP die soon after birth because of kidney failure. In the kidney, CD2AP is expressed in glomerular podocytes, which suggests that it may play a role in a specialized adhesion complex known as the slit diaphragm. One of the major components of the slit diaphragm is nephrin, a podocyte-specific protein. Here we demonstrate that CD2AP localizes to the slit diaphragm in podocytes using immunoelectron microscopy and that nephrin and CD2AP co-immunoprecipitate from a podocyte cell line. The specificity of this interaction was verified by mapping studies, which demonstrated that a novel domain at the C terminus of CD2AP interacts with the C-terminal portion of the nephrin cytoplasmic domain. These studies lend further support to the idea that CD2AP plays a role in the structural integrity of the slit diaphragm. PMID- 11733380 TI - Role of activatory Fc gamma RI and Fc gamma RIII and inhibitory Fc gamma RII in inflammation and cartilage destruction during experimental antigen-induced arthritis. AB - IgG-containing immune complexes, which are found in most RA joints, communicate with hematopoietic cells using three classes of Fc receptors(Fc gamma RI, -II, III). In a previous study we found that if a chronic T-cell-mediated antigen induced arthritis (AIA) was elicited in knee joints of FcR gamma-chain-deficient mice that lack functional Fc gamma RI and Fc gamma RIII, joint inflammation was comparable but severe cartilage destruction was absent. We now examined the individual role of the stimulatory Fc gamma RI and Fc gamma RIII and inhibitory Fc gamma RII in inflammation and functional cartilage damage in knee joints with AIA using Fc gamma RI-, Fc gamma RII-, and Fc gamma RIII-deficient mice. Three weeks after immunization with the antigen-methylated bovine serum albumin (BSA), cellular (T-cell responses as measured by lymphocyte proliferation) immunity raised against mBSA was comparable in all groups examined. Humoral (total IgG, IgG1, IgG2a, and IgG2b levels) immunity against mBSA was comparable in Fc gamma RI-/- and Fc gamma RIII-/- but higher in Fc gamma RII-/- if compared to controls. Joint swelling as measured by (99m)Tc uptake at days 1, 3, and 7 was similar in Fc gamma RI-/- and Fc gamma RIII-/- mice and significantly higher in Fc gamma RII /-. Chronic inflammation and cartilage damage (depletion of proteoglycans, metalloproteinase (MMP)-induced neoepitopes, and matrix erosion) was studied histologically in total knee joint sections stained with hematoxylin or safranin O. Histologically, at day 7 after AIA induction, exudate and infiltrate in the knee joint was similar in Fc gamma RI-/- and Fc gamma RIII-/- and significantly higher (230% and 340%) in Fc gamma RII-/- mice if compared to controls. Aggrecan breakdown in cartilage caused by MMPs and, which is related to severe irreversible cartilage erosion, was further studied by immunolocalization of MMP mediated neoepitopes (VDIPEN) and image analysis. MMP-induced neoepitopes determined in various cartilage layers (tibia and femur) were primarily inhibited in Fc gamma RI-/- (79 to 87% and 87 to 88%, respectively) and comparable in Fc gamma RIII-/-. VDIPEN neoepitopes were much higher (82 to 122% and 200 to 250%, respectively) in Fc gamma RII-/- mice. Initial depletion of proteoglycans was similar (60 to 100%) in all groups. In the chronic phase, cartilage matrix erosion in the lateral and medial tibia was significantly elevated in Fc gamma RII-/- (222% and 186%, respectively) but not in Fc gamma RI-/- or Fc gamma RIII-/ mice. These results suggest that during T-cell-mediated AIA, Fc gamma RI and Fc gamma RIII act in concert in acute and chronic inflammation whereas Fc gamma RI is the dominant FcR involved in severe cartilage destruction. Fc gamma RII is a crucial inhibiting factor in acute and chronic inflammation and cartilage erosion. PMID- 11733381 TI - Embryonic gut anomalies in a mouse model of retinoic Acid-induced caudal regression syndrome: delayed gut looping, rudimentary cecum, and anorectal anomalies. AB - Vitamin A and its derivatives such as retinoic acid (RA) are important signaling molecules for morphogenesis of vertebrate embryos. Little is known, however, about morphogenetic factors controlling the development of the gastrointestinal tract and RA is likely to be involved. In the mouse, teratogenic doses of RA cause truncation of the embryonic caudal body axis that parallel the caudal regression syndrome as described in humans. These changes are often associated with anomalies of the lower digestive tract. Overlapping spatiotemporal expression of retinoic acid receptor-beta (RAR beta) and cellular retinol-binding protein I, CRBPI, with Hoxb5 and c-ret in the gut mesoderm imply possible cooperation required for proper neuromuscular development. To determine susceptibility and responsiveness of the developing gut and its neuromusculature to exogenous retinoids we used a mouse model of RA-induced caudal regression syndrome. The results showed that stage-specific RA treatment both in vivo and in vitro affected gut looping/rotation morphogenesis and growth of asymmetrical structures such as the cecum together with delayed differentiation of the gut mesoderm and colonization of the postcecal gut by neural crest-derived enteric neuronal precursors. These observations demonstrate that RA has a direct effect on gut morphogenesis and innervation. PMID- 11733382 TI - Immunological characterization of human vaginal xenografts in immunocompromised mice: development of a small animal model for the study of human immunodeficiency virus-1 infection. AB - A small animal model for the in vivo study of human immunodeficiency virus-1 and other fastidious infectious agents in human host target tissues is critical for the advancement of therapeutic and preventative strategies. Our laboratory has developed a human vaginal xenograft model that histologically recapitulates features of the human vaginal epithelial barrier. Vaginal xenografts were surgically implanted into C.B.-Igh-1(b)/IcrTac-Prkdc(scid) (SCID) and NOD/LtSz scid/scid (NOD/SCID) mice, with and without human peripheral blood mononuclear cell reconstitution. Immunohistochemical staining of vaginal xenografts demonstrated that in the SCID strain healed vaginal xenografts did not retain intrinsic human immune cells at baseline levels, whereas the NOD/SCID strain supported retention of intrinsic human immune cell populations within the xenografts for at least 2 months after engraftment. In peripheral blood mononuclear cell-reconstituted NOD/SCID mice with vaginal xenografts, flow cytometric analyses detected human immune cell populations in the peripheral blood and immunohistochemical methods detected infiltration of human CD45+ cells in the mouse spleens and vaginal xenografts for at least 2 months after reconstitution. This optimized NOD/SCID human vaginal xenograft model may provide a unique small animal in vivo system for the study of human immunodeficiency virus-1 transmission and infection. PMID- 11733383 TI - Failure of ureteric bud invasion: a new model of renal agenesis in mice. AB - FUBI (failure of ureteric bud invasion) is a highly inbred strain of mouse with a high spontaneous incidence of uni- or bilateral renal agenesis (60%). Bilateral renal agenesis is lethal within 2 days after birth. The primary defect of FUBI is failure of the ureteric bud to penetrate into the metanephric mesenchyme at around embryonic day 11, resulting in apoptosis of metanephric cells and leading to renal agenesis on the affected side. The metanephros seemed to be normal because co-culturing of the FUBI metanephros with homologous spinal cord induced differentiation of the rudiment, but co-culturing with the homologous ureteric bud frequently did not. Genetic analysis revealed that more than two genes were involved in this malformation and we mapped one of the modifier loci, fubi1, on chromosome 2, at approximately 65 cM from the centromere. In this region, there are two possible candidate genes, Wilms' tumor 1 and formin, that play important roles in kidney development. Some of formin mutants shared a similar phenotype with FUBI; however, there was no difference in the expression of formin in embryonic kidneys between FUBI and control NFS/N mice. Studies of fubi1 congenic mice indicated that interaction of two or more loci is essential for the FUBI phenotype. PMID- 11733384 TI - Cryoglobulinemic glomerulonephritis in thymic stromal lymphopoietin transgenic mice. AB - Mixed cryoglobulins are complexes of immunoglobulins that reversibly precipitate in the cold and lead to a systemic disease in humans. Renal involvement usually manifests as a membranoproliferative glomerulonephritis with marked monocyte infiltration and, at times, intracapillary thrombi. Thymic stromal lymphopoietin (TSLP) is a recently cloned cytokine that supports differentiation and long-term growth of B cells. Here we report that TSLP overexpression in mice results in the development of mixed cryoglobulins, with renal involvement closely resembling cryoglobulinemic glomerulonephritis as it occurs in humans. One hundred twenty three mice were sacrificed at monthly intervals, with at least five TSLP transgenic mice and five controls in each group. Blood, kidneys, spleen, liver, lung, and ear were collected and studied by routine microscopy, immunofluorescence, immunohistochemistry, and electron microscopy. TSLP transgenic animals developed polyclonal mixed cryoglobulinemia (type III) and a systemic inflammatory disease involving the kidney, spleen, liver, lung, and ears. Renal involvement was of a membranoproliferative type demonstrating thickened capillary walls with cellular interposition and double contours of the basement membrane, expansion of the mesangium because of increased matrix and accumulation of immune-deposits, subendothelial immune-deposits, focal occlusion of capillary loops, and monocyte/macrophage influx. In contrast to the severe glomerular lesions, the tubulointerstitium was not involved in the disease process. The renal lesions and the disease course were more severe in females when compared to males. We describe a mouse strain in which a B-cell-promoting cytokine leads to formation of large amounts of mixed cryoglobulins and a systemic inflammatory injury that resembles important aspects of human cryoglobulinemia. This is the first reproducible mouse model of renal involvement in mixed cryoglobulinemia, which enables detailed studies of a membranoproliferative pattern of glomerular injury. PMID- 11733385 TI - Immunological consequences of topical bovine thrombin. PMID- 11733390 TI - Will diabetes save the platelet blockers? PMID- 11733391 TI - Protection against oxidized low-density lipoprotein-induced vascular endothelial cell death by integrin-linked kinase. AB - BACKGROUND: Integrin-linked kinase (ILK) is a protein that plays important roles in extracellular matrix-mediated signaling. It has been shown that ILK is expressed preferentially in cardiac and skeletal muscles. Evidence points to its role as an upstream regulator of protein kinase B, a critical player in apoptosis. Because oxidized LDL (oxLDL) is thought to promote atherogenesis by causing the apoptosis of endothelial cells, we investigated the potential roles that ILK may play in oxLDL-induced apoptosis in vascular endothelial cells. METHODS AND RESULTS: Transcriptional and translational levels of ILK were investigated with reverse-transcriptase polymerase chain reaction and Western analysis. oxLDL treatment induced both the transcription and the translation of the ILK gene in endothelial cells. A recombinant adenovirus vector encoding the ILK gene was constructed to investigate its potential role in oxLDL-induced apoptosis in human umbilical vein endothelial cells and mouse lymphoid vein endothelial cells transformed by simian virus 40. In both types of cells, overexpression of the ILK gene significantly prevented oxLDL-induced apoptosis or cell death, as evaluated by 2 independent assay methods. Furthermore, we showed that ILK could inhibit oxLDL-induced upregulation of the kinase activity of p38 mitogen-activated protein kinase, which is often associated with stress-induced pro-apoptotic signal transduction. Finally, examination of other factors, such as bcl-2, bcl-xl, caspase 3, and caspase 9, demonstrated significant changes that were correlated with oxLDL treatment and ILK overexpression. CONCLUSIONS: ILK may be an important factor involved in the regulation of oxLDL-induced apoptosis in vascular endothelial cells. Modifying its activity may be a useful approach for prevention of endothelial cell injury in oxLDL-induced atherosclerosis. PMID- 11733392 TI - Platelet glycoprotein IIb/IIIa inhibitors reduce mortality in diabetic patients with non-ST-segment-elevation acute coronary syndromes. AB - BACKGROUND: Diabetes mellitus is a major risk factor for adverse outcomes after acute coronary syndromes (ACS). Because this disease may be associated with increased platelet aggregation, we investigated whether diabetic patients with ACS derive particular benefit from platelet glycoprotein (GP) IIb/IIIa receptor inhibition. METHODS AND RESULTS: We performed a meta-analysis of the diabetic populations enrolled in the 6 large-scale platelet GP IIb/IIIa inhibitor ACS trials: PRISM, PRISM-PLUS, PARAGON A, PARAGON B, PURSUIT, and GUSTO IV. Among 6458 diabetic patients, platelet GP IIb/IIIa inhibition was associated with a significant mortality reduction at 30 days, from 6.2% to 4.6% (OR 0.74; 95% CI 0.59 to 0.92; P=0.007). Conversely, 23 072 nondiabetic patients had no survival benefit (3.0% versus 3.0%). The interaction between platelet GP IIb/IIIa inhibition and diabetic status was statistically significant (P=0.036). Among 1279 diabetic patients undergoing percutaneous coronary intervention (PCI) during index hospitalization, the use of these agents was associated with a mortality reduction at 30 days from 4.0% to 1.2% (OR 0.30; 95% CI 0.14 to 0.69; P=0.002). CONCLUSIONS: This meta-analysis, including the entire large-scale trial experience of intravenous platelet GP IIb/IIIa inhibitors for the medical management of non-ST-segment-elevation ACS, shows that these agents may significantly reduce mortality at 30 days in diabetic patients. Although not based on a randomized assessment, the survival benefit appears to be of greater magnitude in patients undergoing PCI. Therefore, the use of platelet GP IIb/IIIa inhibitors should be strongly considered in diabetic patients with ACS. PMID- 11733393 TI - Attenuation of rebound ischemia after discontinuation of heparin therapy by glycoprotein IIb/IIIa inhibition with eptifibatide in patients with acute coronary syndromes: observations from the platelet IIb/IIIa in unstable angina: receptor suppression using integrilin therapy (PURSUIT) trial. AB - Background- A reactivation of ischemia after the discontinuation of intravenous heparin in acute coronary syndromes has been described. The effect of glycoprotein IIb/IIIa blockade on heparin rebound is unknown. Methods and Results Patients with acute coronary syndromes who received heparin therapy but not initial revascularization in the Platelet IIb/IIIa in Unstable angina: Receptor Suppression Using Integrilin Therapy (PURSUIT) trial were analyzed. Rates of death or myocardial (re)infarction while on heparin therapy and in 12-hour periods in the 2 days after heparin discontinuation were compared between eptifibatide and placebo. There was no difference between study groups in event rates during heparin infusion. In the 12 hours after heparin discontinuation, there was a 2.5-fold increase in all events, an 8-fold increase in death, and a 2 fold increase in myocardial infarction. However, in the 12 hours after heparin discontinuation, there was a significantly lower rate of events (1.68% versus 2.53%, P=0.03) and death (0.77% versus 0.21%, P=0.002) in the eptifibatide group compared with the placebo group. When only considering patients who were on study drug at the time of heparin discontinuation, the reduction in the combined end point was marginally significant, but the difference in the rate of death remained significant (0.68% versus 0.06%, P=0.004). In logistic regression analyses, the multivariate predictors of rebound events were the duration of heparin therapy, age, North American site, and lack of eptifibatide treatment. Conclusions- An increase in death or myocardial infarction occurs in the 12 hours after heparin discontinuation in patients with acute coronary syndromes. This rebound is attenuated by glycoprotein IIb/IIIa inhibition with eptifibatide. PMID- 11733394 TI - Double-blind, randomized trial of an anti-CD18 antibody in conjunction with recombinant tissue plasminogen activator for acute myocardial infarction: limitation of myocardial infarction following thrombolysis in acute myocardial infarction (LIMIT AMI) study. AB - BACKGROUND: Inhibition of leukocyte adhesion can reduce myocardial infarct size in animals. This study was designed to define the safety and efficacy of a recombinant, humanized, monoclonal antibody to the CD18 subunit of the beta2 integrin adhesion receptors (rhuMAb CD18), in reducing infarct size in patients treated with a thrombolytic agent. METHODS AND RESULTS: The Limitation of Myocardial Infarction following Thrombolysis in Acute Myocardial Infarction Study (LIMIT AMI) was a randomized, double-blind, placebo-controlled, multicenter study conducted in 60 centers in the United States and Canada. A total of 394 subjects who presented within 12 hours of symptom onset with ECG findings (ST-segment elevation) consistent with AMI were treated with recombinant tissue plasminogen activator and were also given an intravenous bolus of 0.5 or 2.0 mg/kg rhuMAb CD18 or placebo. Coronary angiography was performed at 90 minutes, 12-lead ECGs were obtained at baseline, 90, and 180 minutes, and resting sestamibi scans were performed at >/=120 hours. Adjunctive angioplasty and use of glycoprotein IIb/IIIa antiplatelet agents at the time of angiography were discretionary. There were no treatment effects on coronary blood flow, infarct size, or the rate of ECG ST-segment elevation resolution, despite the expected induction of peripheral leukocytosis. A slight trend toward an increase in bacterial infections was observed with rhuMAb CD18 (P=0.33). CONCLUSIONS: RhuMAb CD18 was well tolerated but not effective in modifying cardiac end points. PMID- 11733395 TI - Collateral function in chronic total coronary occlusions is related to regional myocardial function and duration of occlusion. AB - BACKGROUND: Collateral circulation can maintain myocardial function and viability in chronic total coronary occlusion (TCO). The present study evaluates the relation of myocardial function and duration of occlusion to collateral function. METHODS AND RESULTS: A total of 50 patients underwent a successful recanalization of a TCO (>4 weeks' duration). Collateral function was assessed by intracoronary Doppler and pressure recordings before the first balloon inflation and after PTCA had been completed. Collateral function was assessed by Doppler- (CFI(D)) and pressure-derived collateral flow indices (CFI(P)), as well as indices of collateral (R(Coll)) and peripheral resistance (R(P)). Patients with normokinesia had lower R(Coll) (4.9+/-2.5 versus 11.8+/-8.2 mm Hg. cm(-1). s(-1); P=0.033) and lower R(P) (3.8+/-1.9 versus 6.1+/-4.1 mm Hg. cm(-1). s(-1); P=0.031) than those with akinesia. Patients with akinesia and a TCO duration of /=10 000 micrometer(2)) were found in 48 aspirates (83%). The median number of particles, their maximum diameter, and their maximum area were all significantly higher in the aspirates obtained during procedures associated with PNCs than in aspirates obtained during procedures not associated with PNCs. However, pronounced overlap in the distributions (PNCs versus no PNCs) of the number and maximum diameter of particles precluded any predictive inferences. In contrast, a maximum particle area >800 000 micrometer(2) (>0.8 mm(2)) was associated with a 60% chance of having a PNC. CONCLUSIONS: Despite balloon protection, PNCs occurred in 5.2% of patients who underwent CAS procedures. The maximum area of aspirated particles seems to be an indicator of increased risk for PNCs. PMID- 11733397 TI - Clinical correlates and reference intervals for pulmonary artery systolic pressure among echocardiographically normal subjects. AB - BACKGROUND: Data in normal human subjects on the factors affecting pulmonary artery systolic pressure (PASP) are limited. We determined the correlates of and established a reference range for PASP as determined by Doppler transthoracic echocardiography (TTE) from a clinical echocardiographic database of 102 818 patients, of whom 15 596 (15%) had a normal Doppler TTE study. METHODS AND RESULTS: A normal TTE was based on normal cardiac structure and function during complete Doppler TTE studies. The PASP was calculated by use of the modified Bernoulli equation, with right atrial pressure assumed to be 10 mm Hg. Among TTE normal subjects, 3790 subjects (2432 women, 1358 men) from 1 to 89 years old had a measured PASP. The mean PASP was 28.3+/-4.9 mm Hg (range 15 to 57 mm Hg). PASP was independently associated with age, body mass index (BMI), male sex, left ventricular posterior wall thickness, and left ventricular ejection fraction (P<0.001). The estimated upper 95% limit for PASP among lower-risk subjects was 37.2 mm Hg. A PASP >40 mm Hg was found in 6% of those >50 years old and 5% of those with a BMI >30 kg/m(2). CONCLUSIONS: Among 3790 echocardiographically normal subjects, PASP was associated with age, BMI, sex, wall thickness, and ejection fraction. Of these subjects, 28% had a PASP >30 mm Hg, and the expected upper limit of PASP may include 40 mm Hg in older or obese subjects. These findings support the use of age- and BMI-corrected values in establishing the expected normal range for PASP. PMID- 11733398 TI - Radiofrequency catheter ablation in infants 60% were overestimating their physical activity level. Furthermore, respondents who were not aware of their inadequate physical activity level had a less positive intention to increase their level of physical activity than those who rated their own physical activity level as low. Respondents in the pre-contemplation and maintenance stages had a less positive attitude, perceived less social support and had lower self-efficacy expectations towards increasing physical activity than those in the contemplation, preparation and action stages. The results suggest that interventions aimed at increasing physical activity in the studied population should be aimed at increasing awareness of personal activity levels and should be stage-matched. PMID- 11733450 TI - Different worlds, different tasks for health promotion: comparisons of health risk profiles in Chinese and Finnish rural people. AB - The aim of this study was to compare cardiovascular risk factors of working-aged people in Chinese and Finnish rural villages. The surveys were carried out in 1989 in Tianjin, China, and in Kuopio, Finland. Altogether, 897 Chinese inhabitants and 795 Finnish subjects participated in the surveys. Health behaviours were recorded, and height, weight, blood pressure, heart rate and serum lipids were measured. Generally Finns had a significantly higher mean body mass index, systolic and diastolic blood pressures, and serum total cholesterol, low-density lipoprotein cholesterol, and total cholesterol/high-density lipoprotein ratio than the Chinese. However, no difference was seen between Chinese and Finnish women in diastolic blood pressure and serum triglycerides. Lower high-density lipoprotein cholesterol levels were observed in Finnish men than in Chinese men, whereas a higher mean level was shown in Finnish women than in Chinese women. There were significantly higher mean heart rates and prevalence of smoking in Chinese than in Finnish populations. More people who were overweight, obese and hypertensive were found in the Finnish than in the Chinese populations. Most of the Finns had two or more cardiovascular risk factors compared with the Chinese, the majority of whom were in the group with less than two risk factors. In conclusion, the risk profiles are clearly somewhat different in these two countries. A major task for the Chinese health policy and health care system is to decrease smoking and to prevent obesity and hypertension. In Finland, the biggest task seems to be the reduction of weight and lipid abnormalities, and the prevention of hypertension. PMID- 11733451 TI - School and anaemia prevention: current reality and opportunities--a Tanzanian case study. AB - Iron deficiency anaemia is highly endemic in rural areas of Tanzania and in many developing countries. Its prevention among school children requires greater dissemination of knowledge of anaemia among children, teachers, parents and the general community. Associated improvements in the hygienic status of domestic and school environments are also often required. One-hundred-and-thirty-one anaemic children, 90 parents and 76 teachers were interviewed to ascertain their understanding of anaemia. Most children and parents had little knowledge of the symptoms, causes and prevention of anaemia. In addition to their iron-deficient diets, more than half of the children went to school without something to eat at breakfast and during school hours. However, parents and teachers were willing to work together to provide meals for the children. Poor sanitation in the children's homes and in schools was a little recognized factor which could pose a serious risk of anaemia. In addition, inadequate sanitation facilities and poor quality of physical environment prevailed both in the children's homes and in schools. The findings suggest the need for the establishment of a health promoting schools network to provide a comprehensive framework for health promotion in schools as well as in homes in Tanzania and in other developing countries. Schools can be an ideal setting to positively influence a community's health status. Partnerships among teachers, parents and the wider community are required to identify, prioritize and ameliorate health problems. PMID- 11733452 TI - Paid celebrity endorsement in health promotion: a case study from Australia. AB - In late 1998, Australian cricketer Shane Warne was allegedly paid A$200 000 ( pound78 060, US$123 000) by a pharmaceutical company to publicize his attempt to stop smoking. Warne failed to stop, and his continuing smoking remained newsworthy more than a year later. The arrangement caused considerable media controversy about the ethics of payment for charitable or socially worthy actions. This paper explores the community's reaction to payment for modelling a healthy behaviour, discussing the values that Warne transgressed and whether these mattered, given that the campaign caused an unprecedented rise in the use of nicotine replacement therapy. PMID- 11733453 TI - 'Settings' based health promotion: a review. AB - Over the past 10 years, 'settings' based health promotion has become a central feature of efforts to promote health that recognize the significance of context. Emerging in part from a perception of an over-reliance on individualistic methods, the approach was built on a profound belief in its value and deployed a range of novel theoretical resources, mainly from organizational sociology and psychology. This initial enthusiasm has been maintained within policy directives, in the published literature and, from our own experience, amongst health promotion practitioners. At the same time, with the maturing of the approach, has come a healthy element of critical review. Drawing upon the literature and based upon our experiences within the Health Education Board for Scotland, this paper seeks to bring together a range of perspectives, casting a critical yet constructive eye on current settings theory and practice. The paper first reviews the nature of settings based work, highlighting the varied bases and expectations that underpin it. Similarly, the many factors that influence the ability of health promoters to deliver such activities are considered. In relation to the construction and delivery of such activity, the paper suggests that there needs to be an explicit and detailed assessment of the nature of the setting, the skills of the health promoter and the associated expectations. PMID- 11733454 TI - Determinants of smoking and cessation during and after pregnancy. AB - Smoking during pregnancy is harmful to both the foetus and the woman herself. However, in spite of educational efforts, a substantial proportion of pregnant women continue to smoke and many women who do stop smoking during pregnancy resume smoking following childbirth. To foster successful maternal smoking cessation, public health professionals need to focus on the major determinants of smoking and cessation during and after pregnancy, and then to address these with their intervention efforts. It is important to review contemporary epidemiological evidence on this significant public health issue. We have identified nine cohort studies, published in international peer-reviewed journals, that have examined determinants of smoking and cessation in pregnant women. The results indicate that the determinants of pregnant smoking and cessation include maternal age, dose and duration of smoking, partner's smoking habit, socioeconomic status, level of education, age to start smoking, level of addiction, parity and passive smoking. However, many other psychosocial factors, which may affect smoking status among pregnant women, remain to be identified. Evidence reviewed here suggests that a more focused, integrated approach and a more comprehensive assessment of major determinants of smoking and cessation during pregnancy will be required as part of any future intervention effort. PMID- 11733455 TI - Setting standards in the evaluation of community-based health promotion programmes--a unifying approach. AB - Community-based health promotion often emphasizes elements of empowerment, participation, multidisciplinary collaboration, capacity building, equity and sustainable development. Such an emphasis may be viewed as being in opposition to equally powerful notions of evidence-based decision making and accountability, and with funders' and government decision-makers' preoccupation with measuring outcomes. These tensions may be fuelled when community practitioners and lay participants feel evaluations are imposed upon them in a manner that fails to appreciate the uniqueness of their community, its programme, and practitioners' skills and experience. This paper attempts to provide an approach that depicts evaluation as being mutually beneficial to both funders/government and practitioners. First, a values stance for health promotion, termed a 'salutogenic' orientation, is proposed as a foundation for the evaluation of community-based health promotion. Secondly, we discuss possible objects of interest, the first component of an evaluation. We then discuss the spirit of the times and its implications for community-based health promotion. Finally, we address the key question of setting standards. A typology of standards is presented. Arbitrary, experiential and utility standards are based on perceived needs and priorities of practitioners, lay participants or professional decision makers. Historical, scientific and normative standards are driven by empirical, objective data. Propriety and feasibility standards are those wherein the primary concern is for consideration of resources, policies, legislation and administrative factors. The 'model' standards approach is presented as an exemplar of a combined approach that incorporates elements of each of the other standards. We argue that the 'optimal' standard for community-based health promotion depends on the setting and the circumstances. There is no 'magic bullet', 'one-size-fits-all' or 'best' standard. Further, we argue that standards should be set from an inclusive, salutogenic orientation. This approach offers a means of creating a situation in which policy-makers and funders are more supportive of evaluation designs that fit with community realities, and community stakeholders are more capable and consistent in rigorously evaluating community based health promotion programmes and policies. PMID- 11733456 TI - Opportunities to improve maternal health literacy through antenatal education: an exploratory study. AB - This paper examines the concept of maternal health literacy, defined as the cognitive and social skills that determine the motivation and ability of women to gain access to, understand, and use information in ways that promote and maintain their health and that of their children. Specifically, it investigates the feasibility of using the concept of health literacy to guide the content and process of antenatal classes. The paper reports on the results of focus groups and interviews conducted with a range of health care providers, pregnant women and new mothers to obtain different perspectives on the issues surrounding antenatal education and parenting. The results give us a realistic look at what women are learning from existing antenatal education and how it can be improved. Comparing the results from the educators and the women, the same basic issues surface. Both recognize that there are serious time limitations in antenatal classes. These limitations, combined with natural anxiety and curiosity about childbirth, generally ensure that the content of classes is confined to pregnancy and childbirth. The limitations of time are also cited as a reason for the teaching methods being heavily weighted towards the transfer of factual information, as distinct from the development of decision-making skills, and practical skills for childbirth and parenting The results indicate clearly that antenatal classes cannot possibly cover all there is to know about pregnancy, childbirth and parenting. If the purpose of antenatal classes is to improve maternal health literacy, then women need to leave a class with the skills and confidence to take a range of actions that contribute to a successful pregnancy, childbirth and early parenting. This includes knowing where to go for further information, and the ability to analyse information critically. The authors conclude that this would represent a very challenging change in orientation for both the educators and pregnant women included in this study. Work continues on the development of the tools that will be needed to support this change. PMID- 11733457 TI - Imiquimod and resiquimod as novel immunomodulators. AB - Augmenting the host's natural immune response to viruses by the administration of exogenous cytokines such as interferon-alpha (IFN-alpha) is a strategy increasingly employed in antiviral therapeutics. Enhancing the release of endogenous cytokines is, however, an alternative approach. The imidazoquinolinamines imiquimod and resiquimod have demonstrated potency as inducers of IFN-alpha and other cytokines both in vitro and in vivo. Cytokine gene activation is mediated via the signal transducer and activator of transcription 1 (STAT-1) and involves the transcription factors NFkappaB and alpha4F1. Antiviral activity has been demonstrated against a variety of viruses, and clinical efficacy has been demonstrated against genital warts, herpes genitalis and molluscum contagiosum. Imiquimod is administered as a 5% cream (Aldara) and has been licensed for the treatment of anogenital warts in immunocompetent patients. Complete clearance of warts has been observed in up to half of treated patients with only local side effects reported. Resiquimod can be administered topically but also exists as an oral formulation. The range of potential infections for which these agents may have clinical utility includes chronic hepatitis C virus infection and Kaposi's sarcoma. In addition, the imidazoquinolinamines may find roles in the therapy of cancers and as vaccine adjuvants. PMID- 11733458 TI - Novel non-nucleoside inhibitors of cytomegaloviruses (BAY 38-4766): in vitro and in vivo antiviral activity and mechanism of action. AB - For two decades it has been impossible to develop drugs with novel mechanisms of action against herpesviruses, and treatment has been confined largely to the use of inhibitors of viral DNA polymerase. As a representative of a novel inhibitory approach, the non-nucleosidic BAY 38-4766 was identified as a highly selective inhibitor of human cytomegalovirus (HCMV). The compound selectively inhibits not only HCMV strains, including ganciclovir-resistant, ganciclovir/foscarnet and ganciclovir/cidofovir double-resistant clinical isolates, but also a number of monkey and rodent cytomegaloviruses. In a murine cytomegalovirus (MCMV) pathogenicity model in mice, antiviral efficacy and excellent tolerability were demonstrated. BAY 38-4766-resistant HCMV and MCMV strains are not cross-resistant to the nucleoside analogues ganciclovir and cidofovir or the pyrophosphate analogue foscarnet, indicating a different mode of action. Mechanistic studies demonstrated that the high selectivity of this drug class is most likely due to the inhibition of a late stage of the viral replication cycle. Sequence analyses of resistant HCMV and MCMV strains revealed mutations in UL89 and UL104, proteins known to be involved in viral DNA cleavage and packaging. Consequently, the drug is highly specific for the viral as opposed to cellular functions, since UL89 is related to a bacteriophage terminase and no human equivalent exists. In addition, because some of the genes of the viral DNA cleavage and packaging complex are highly conserved among herpesviruses, development of broad-spectrum agents covering additional human herpesviruses might be possible using this approach. PMID- 11733459 TI - Activity of the new antifungal triazole, posaconazole, against Cryptococcus neoformans. AB - The new antifungal derivative posaconazole was tested against three clinical isolates of Cryptococcus neoformans var. neoformans using a broth microdilution procedure performed according to the guidelines established by the NCCLS. Posaconazole MICs were 0.125, 0.25 and 1.0 mg/L for isolates 491, 2337 and 486, respectively. To investigate the in vivo activity of this new compound, we established an experimental model of systemic cryptococcosis in CD1 mice by iv injection of cells of each strain of C. neoformans. Low (3 mg/kg/day) and high (10 mg/kg/day) doses of posaconazole were compared with amphotericin B given at 0.3 mg/kg/day for 10 consecutive days. Survival studies showed that all treatment regimens were effective in prolonging the survival of mice infected with C. neoformans 486 (P < 0.001). Only posaconazole at 10 mg/kg and amphotericin B were effective in prolonging the survival in mice infected with C. neoformans 2337 (P from <0.01 to <0.001), while neither agent was effective in mice infected with C. neoformans 491. Tissue burden experiments performed 24 h after the end of therapy revealed that posaconazole at 10 mg/kg was effective at reducing the fungal burden in both lung and brain tissues of all three strains of C. neoformans. In particular, for C. neoformans 491 and 2337 posaconazole was superior to amphotericin B at reducing the fungal burden in the brain (P < 0.05). The efficacy of posaconazole was also confirmed by determining the capsular antigen serum levels of treated mice versus untreated mice. Our study underlines the excellent activity of posaconazole against this pathogenic yeast. PMID- 11733460 TI - Antifungal properties of selective serotonin reuptake inhibitors against Aspergillus species in vitro. AB - This study investigated the fungicidal activity of selective serotonin reuptake inhibitors (SSRIs) against clinical isolates of Aspergillus fumigatus (n = 11), Aspergillus flavus (n = 9), Aspergillus terreus (n = 10) and Candida parapsilosis (ATCC 22019). The common drugs fluoxetine, seroxate, sertraline, paroxetine and reboxetine were applied in a broth microdilution test. In addition, we examined whether a post-antibiotic effect occurs following short exposure to the drugs. The various SSRIs showed time- and dose-dependent effects and were fungicidal towards the organisms tested. Sertraline and fluoxetine were the most active drugs, yet there were differences in the susceptibility of the various isolates tested. A lag of regrowth was dependent on the various SSRIs tested and their concentration. Treatment for 4 h at concentrations of sertraline below and equipotent to the minimal fungicidal concentration resulted in a lag of regrowth of 8-24 h for isolates of A. fumigatus and A. flavus. In conclusion, our in vitro studies clearly demonstrate antifungal effects of SSRIs. Animal studies are needed to evaluate the potential role of these psychotropic drugs in the management of fungal infections. PMID- 11733461 TI - Bumetanide annihilation of amphotericin B-induced apoptosis and cytotoxicity is due to its effect on cellular K+ flux. AB - The antifungal antibiotic amphotericin B causes considerable toxic effects during clinical therapy. We have shown previously that amphotericin B-induced cytotoxicity and apoptosis were eradicated by the Na+, K+, 2Cl- cotransport inhibitor bumetanide. To elucidate the role of K+ flux and the activity of Na+, K+ ATPase and Na+, K+, 2Cl- cotransport in apoptosis and cytotoxicity induced by amphotericin B alone and combined with bumetanide, we quantified the influx and efflux of K+ of mesothelioma cells (P31) using the K+ analogue 86Rb+ with ouabain (100 micromol/L) as the K+ influx probe. To determine the susceptibility of Candida albicans to amphotericin B when combined with bumetanide we used a plate diffusion method. Amphotericin B or bumetanide alone significantly stimulated 86Rb+ efflux during the first 15 min. However, when added simultaneously, the cellular 86Rb+ efflux was markedly decreased. Amphotericin B (3 mg/L) had no effect on immediate (15 min) total 86Rb+ influx. When bumetanide (100 micromol/L) was added, the total 86Rb+ influx was markedly reduced due to inhibition of augmented Na+, K+, 2Cl- cotransport and low Na+, K+ ATPase activity. Bumetanide did not affect the susceptibility of C. albicans to amphotericin B, which suggests that bumetanide or related drugs could be used in antifungal therapy to increase amphotericin B effectiveness without increasing its adverse effects. We suggest that bumetanide hampering of amphotericin B-induced cytotoxicity and apoptosis could be due to an immediate reduction of cellular K+ efflux as well as disordered K+ influx. PMID- 11733462 TI - Accumulation of five fluoroquinolones by Mycobacterium tuberculosis H37Rv. AB - The accumulation of ciprofloxacin, norfloxacin, moxifloxacin, levofloxacin and ofloxacin by Mycobacterium tuberculosis H37Rv was determined with a modified fluorescence method. The time to achieve a steady-state concentration (SSC) of each agent in M. tuberculosis was 60-240 s. Moxifloxacin was accumulated to the lowest concentration and ciprofloxacin to the highest. However, ciprofloxacin took longer to achieve an SSC than the other four agents; levofloxacin reached steady state in the shortest time. Larger fluoroquinolones accumulated to the lowest concentration and more slowly. Although all five agents had low hydrophobicity values (P(app) < or =0.11), those with the lowest values accumulated to the higher concentrations. PMID- 11733463 TI - Synergy of different antibiotic combinations in biofilms of Staphylococcus epidermidis. AB - The in vitro effect of nine antibiotic combinations was investigated in Staphylococcus epidermidis biofilms using ATP-bioluminescence for viable bacterial cell quantification. Four slime-producing (SP) strains were used to form biofilms 6, 24 and 48 h old. These biofilms were exposed for 24 h to antibiotics at 4 x, 2 x, 1 x and 0.5 x MIC. Combinations involving tetracycline together with another antibiotic were the most efficient at the biofilm age and concentration range under study. The combination vancomycin-rifampicin produced the highest bactericidal effect on 6 h biofilms at 4 x MIC, but this effect decreased dramatically in older biofilms. To detect possible antibiotic synergy in combinations that had a significant killing effect, antibiotics were studied not only in combination but also individually. Synergic effects were observed in all the combinations tested. Differences between the effect in combination and the sum of individual antibiotic effects (degree of synergy) were significant (mostly P< 0.001) and exceeded 1 log10 cfu/mL in the majority of cases. In 48 h biofilms, antibiotics caused a significant bactericidal effect when applied in combination, but never when used individually. These results indicate that the biofilm test applied allows the detection of synergy between antibiotics and suggests that this assay could be useful in clinical and extensive synergy studies on S. epidermidis biofilms. PMID- 11733464 TI - Natural antimicrobial susceptibilities of Plesiomonas shigelloides strains. AB - The natural susceptibility of 74 Plesiomonas shigelloides strains isolated from humans (n = 50), water (n = 22) and animals (n = 2) to 71 antibiotics was examined. MICs were performed using a microdilution procedure with Mueller-Hinton broth and an inoculum of 1 x 10(6) cfu/mL. Plesiomonas strains were naturally susceptible or naturally susceptible and intermediate to tetracyclines, several aminoglycosides, aminopenicillins in combination with beta-lactamase inhibitors, all cephalosporins except cefoperazone, ceftazidime and cefepime, carbapenems, aztreonam, quinolones, trimethoprim, sulfamethoxazole, azithromycin, chloramphenicol, nitrofurantoin and fosfomycin. Uniform natural resistance was found to all penicillins tested, roxithromycin, clarithromycin, lincosamides, streptogramins, glycopeptides and fusidic acid. Plesiomonas strains were naturally resistant and intermediate to streptomycin, erythromycin and rifampicin. There were two susceptibility patterns to piperacillin/tazobactam, several cephalosporins and aztreonam. In contrast to a previous study with beta lactam antibiotics, susceptibility testing of non-beta-lactams revealed no alterations of the MICs of most antibiotics, using different inocula and media. A database is described of the natural susceptibility of P. shigelloides strains to a wide range of antibiotics. It can be used for the validation of susceptibility test results of these bacteria. PMID- 11733465 TI - A pharmacokinetic/pharmacodynamic approach to show that not all fluoroquinolones exhibit similar sensitivity toward the proconvulsant effect of biphenyl acetic acid in rats. AB - The proconvulsant effect of biphenyl acetic acid (BPAA) on several fluoroquinolones (FQs) was investigated in vivo, by measuring drug concentrations in the biophase at the onset of convulsions. Male Sprague-Dawley rats (n = 134) were given BPAA orally, at various doses 1 h before starting FQ infusion, which was maintained until the onset of maximal seizures, when cerebrospinal fluid (CSF) and plasma samples were collected for drug concentration determination. The FQ-BPAA interactions in the biophase (CSF) were adequately described on most occasions by an inhibitory Emax effect model with a baseline effect parameter. The efficacy of the proconvulsant effect was characterized by the ratio of the CSF concentrations of FQs at the onset of convulsant activity when BPAA was absent (CCSF0, FQs) and as BPAA CSF concentrations tended toward infinity (CCSFbase, FQs). This ratio varied from 15 for enoxacin to 1.9 for sparfloxacin. The potency of the proconvulsant effect was characterized by the CSF concentration of BPAA corresponding to a proconvulsant effect half of its maximum. This parameter varied between 0.18 +/- 0.06 micromol/L with enoxacin and 15.0 +/- 12.1 micromol/L with sparfloxacin. The CSF diffusion of all FQs was apparently non-linear, as well as the plasma protein binding of BPAA, complicating interpretation of plasma data. The important variability in the proconvulsant effect of BPAA demonstrated in this study between various FQs suggests that in vitro gamma-aminobutyric acid (GABA) binding experiments conducted in the presence of BPAA are unlikely to be good predictors of FQ convulsant risk in clinical practice. PMID- 11733466 TI - The pharmacodynamics of gatifloxacin and ciprofloxacin for pneumococci in an in vitro dynamic model: prediction of equiefficient doses. AB - Enhanced activity against Streptococcus pneumoniae is one of the putative advantages of gatifloxacin over older fluoroquinolones such as ciprofloxacin. This study examined ciprofloxacin and gatifloxacin pharmacodynamics against two differentially susceptible clinical isolates of S. pneumoniae (gatifloxacin MIC, 0.125 and 2 mg/L; ciprofloxacin MIC, 1 and 32 mg/L). The pharmacokinetics of gatifloxacin (single dose) and ciprofloxacin (two 12 hourly doses) with half lives of 6 and 5 h, respectively, were simulated using a two-compartment dynamic model. The AUC/MIC ratios in the peripheral compartments that contain bacterial cultures varied over a four- to five-fold range, from 11 to 48 h with ciprofloxacin and from 15 to 78 h with gatifloxacin. The intensity of the antimicrobial effect (IE) increased with increasing AUC/MIC ratios in a strain independent fashion, although different relationships of IE to log AUC/MIC were inherent for each drug (r2 0.73 for gatifloxacin and r2 0.94 for ciprofloxacin). Subsequently, the respective dose-response relationships of gatifloxacin and ciprofloxacin for a hypothetical strain of S. pneumoniae with MIC equal to the MIC50 were modelled. Based on these relationships, the equiefficient doses of gatifloxacin and ciprofloxacin were predicted for MIC50S of 0.4 and 1 mg/L, respectively. Gatifloxacin 400 mg was predicted to be equiefficient to ciprofloxacin 1400 mg. To provide the same anti-pneumococcal effect as the usual 1000 mg daily dose of ciprofloxacin, the respective daily dose of gatifloxacin could be as low as 180 mg. This in vitro study demonstrates advantages of gatifloxacin relative to ciprofloxacin in terms of the dose-dependent total antimicrobial effect. PMID- 11733467 TI - Vibrio cholerae O1 outbreak isolates in Mozambique and South Africa in 1998 are multiple-drug resistant, contain the SXT element and the aadA2 gene located on class 1 integrons. AB - The characteristics of Vibrio cholerae O1 biotype El Tor, serotype Ogawa isolates from outbreaks of cholera in 1998 amongst migrant workers in the South African provinces of Gauteng and Mpumalanga, on the border of Mozambique, are reported. The isolates seem to have originated from the same clone since they are of two closely related BglI ribotypes. These ribotypes had a high similarity to ribotypes of V. cholerae O1 recently found in three South-east Asian countries. Isolates were resistant to furazolidone, streptomycin, sulfamethoxazole, trimethoprim and tetracycline. Only two isolates contained plasmids of 54 and 63 kb in size. PCR and DNA sequencing revealed that the chromosomally located resistance determinants present included an aadA2 gene cassette contained in a class 1 integron; the SXT element, which is a transposon-like element containing resistance genes; and the tetA gene. A co-transfer of chromosomal closely located genes encoding the SXT element and tetA was shown by mating experiments, PCR and pulsed-field gel electrophoresis analyses. Our study shows for the first time that multiple-resistant V. cholerae O1 isolates containing class 1 integrons and the SXT element were responsible for cholera outbreaks in Southern Africa. Studies are needed to determine the spread of this multiple-resistant O1 strain and further genetic details of the association of the SXT element, tetA and class 1 integrons, including their means of transfer. PMID- 11733468 TI - SHV-12, SHV-5, SHV-2a and VEB-1 extended-spectrum beta-lactamases in Gram negative bacteria isolated in a university hospital in Thailand. AB - Sixty-one extended-spectrum beta-lactamase (ESBL)-producing isolates were collected from Srinagarind Hospital, Thailand. These included 43 Enterobacteriaceae and 18 Pseudomonadaceae. The 43 Enterobacteriaceae were found to produce the following ESBLs: 26 (60.5%) SHV-12, 13 (30.2%) SHV-5, two (4.7%) SHV-2a, one (2.3%) VEB-1 and one (2.3%) unidentified. Twenty-four isolates (55.8%) also carried bla(TEM-1B), as well as bla(SHV) or bla(VEB-1). Plasmid DNA from transconjugants carrying the bla(SHV-12) gene showed various restriction patterns, indicating the distribution of the bla(SHV-12) gene among different antibiotic resistance plasmids. In contrast, bla(SHV-5) in 13 isolates was found on a single plasmid of c. 130 kb. Pulsed-field gel electrophoresis (PFGE) analysis of genomic DNA from these isolates revealed that nine of 11 Klebsiella pneumoniae gave the same pattern, indicating clonal spread of the strain within the hospital, together with the occasional spread of the plasmid to other strains. Among the pseudomonad isolates, 16 Pseudomonas aeruginosa and one Pseudomonas putida had bla(VEB-like) and one P. aeruginosa had bla(SHV-12). Nine of the 16 isolates carrying bla(VEB-like) (56.3%) had identical PFGE patterns, suggesting the dissemination of this gene, also by clonal spread. At least six different bla(VEB-like-)containing integrons were found among the 18 isolates. This is the first report of bacteria producing SHV-12 and SHV-2a in Thailand and the first report of SHV-12 in P. aeruginosa, of VEB-1 in Citrobacter freundii and a VEB-1-like beta-lactamase in P. putida. These findings indicate that ESBL genes in the Far East are part of a gene pool capable of broad horizontal gene transfer, in that these genes can transfer between different families of Gram negative bacilli. PMID- 11733469 TI - Evolution in the trends of antimicrobial resistance in Neisseria gonorrhoeae isolated in Durban over a 5 year period: impact of the introduction of syndromic management. AB - Antimicrobial susceptibility testing was performed on isolates of Neisseria gonorrhoeae obtained from patients attending the City Health STD clinic in Durban, KwaZuluNatal, using the following drugs: penicillin, tetracycline, ciprofloxacin, ofloxacin, ceftriaxone, spectinomycin, erythromycin and azithromycin. These isolates were collected over a 6 year period from 1995 to 2000. Four hundred and fifteen strains were tested: 61 in 1995, 198 in 1997, 98 in 1998/99 and 58 in 1999/2000. A shift to the right is observed in the susceptibilities of N. gonorrhoeae to the currently recommended drugs in the syndromic management guidelines viz. penicillin, tetracycline, ceftriaxone, ciprofloxacin, spectinomycin and erythromycin. The prevalence of penicillinase producing N. gonorrhoeae is currently c. 30%, whereas that of plasmid-mediated tetracycline-resistant N. gonorrhoeae is c. 50%. There is a definite association between the MICs of strains falling within the penicillin and tetracycline chromosomally resistant group, and strains exhibiting a decreased susceptibility to ciprofloxacin and ceftriaxone. The MICs of azithromycin showed a similar distribution when compared with erythromycin for 1999/2000 isolates. We postulate that the presence of efflux pumps might play a role in the increasing MICs that we observe among structurally unrelated groups of drugs. Furthermore, widespread use of these antimicrobials in the community may offer a selective advantage to the development of resistance. The implications of this are far reaching and the local susceptibility trends of N. gonorrhoeae need to be monitored constantly to direct therapy. PMID- 11733470 TI - Tetracycline delivery from fibrin controls peritoneal infection without measurable systemic antibiotic. AB - The addition of antibiotics to an adhesive haemostat results in an ideal system for the treatment of a localized infectious disease. Fibrin sealant (FS) is a biocompatible, resorbable, adherent haemostat that can deliver antibiotics. Previous use of fibrin to deliver antibiotics resulted in rapid release and limited bioactivity. We have reported previously that poorly soluble antibiotics significantly retard release from FS, resulting in extended delivery in vitro, and overcome antibiotic-resistant infection. We now report that localized antibiotic delivery from FS controls peritoneal infection without measurable systemic antibiotic. Rats and mice were implanted with preformed FS discs containing tetracycline free-base to evaluate control of peritoneal sepsis and to measure serum tetracycline levels. Infection was initiated with Staphylococcus aureus. Morbidity and mortality were evaluated for 14 days. Serum was isolated from jugular vein blood with subsequent evaluation for antimicrobial activity. Mice prophylactically treated with FS-tetracycline (FS-TET) 500 mg/kg 2 days before infection cleared the S. aureus infection, resulting in 100% survival. Mice treated with FS-TET 500 mg/kg 7 days before infection survived. Mice treated with FS-TET 1750 mg/kg 35 days before infection also survived. Rats treated with FS-TET 500 mg/kg had undetectable serum tetracycline levels, whereas in vitro release of tetracycline from FS-TET pellets in rat serum was readily detected. We conclude that fibrin is an excellent vehicle for extended delivery of low solubility tetracycline. Tetracycline delivered from FS is an appropriate chemotherapy for S. aureus peritonitis. FS-TET controls localized infection without a measurable concentration of systemic tetracycline. PMID- 11733472 TI - The early bactericidal activity of a low-clearance liposomal amikacin in pulmonary tuberculosis. AB - The early bactericidal activity (EBA) of a liposomal preparation of amikacin (MiKasome) with a long plasma half-life of 120-200 h was examined in seven patients with newly diagnosed, smear-positive pulmonary tuberculosis. Liposomal amikacin was given in slow iv infusions of 30 mg total amikacin/kg body weight on three successive days. Cfu counts were set up on 16 h sputum collections preceding the first dose and following each dose and were used for calculating the EBA. Despite the high concentrations of total amikacin, >1000 mg/L, obtainable in plasma, no evidence of EBA was obtained. In view of the considerable activity of liposomal amikacin in experimental murine tuberculosis, this finding indicates that liberation of amikacin from the long-life liposomes occurs only in macrophages that are not usually present in the vicinity of the large extracellular clumps of bacilli in the cavity caseum. PMID- 11733471 TI - The activity of low-clearance liposomal amikacin in experimental murine tuberculosis. AB - Most of the amikacin in low-clearance liposomal amikacin is excreted very slowly, offering the possibility of maintaining effective treatment of pulmonary tuberculosis with widely separated supervised doses. As a preliminary to explorations in humans, its efficacy was assessed in acute experimental murine tuberculosis by weekly counts of viable bacilli in spleen and lungs over a 4 week period. Liposomal amikacin in dosages of 160, 80 and 40 mg/kg given iv three times a week was 2.4-5.0 times more active than free amikacin and 6.6-6.7 times more active than streptomycin with the non-liposomal drugs given im five times a week. When the free amikacin and the streptomycin were also given iv three times a week, liposomal amikacin was 2.7-2.9 times more active than free amikacin and 3.7-5.6 more active than streptomycin. In a model of chronic tuberculosis, initial BCG vaccination was followed by challenge with virulent Mycobacterium tuberculosis and a 2 week stabilization period. Thereafter, treatment with liposomal amikacin 160 and 80 mg/kg three times a week for the first 4 weeks and then once a week for a further 4 weeks, had greater initial bactericidal activity than free amikacin 160 mg/kg five times a week, but had less eventual sterilizing activity than five times a week oral isoniazid 25 mg/kg or rifampicin 15 mg/kg. Although low-clearance liposomes increased the safety, potency and dosing interval of amikacin in these models, all aminoglycosides, including liposomal amikacin, were only bactericidal in the presence of bacillary metabolism and growth. PMID- 11733473 TI - Elimination of the piperacillin/tazobactam combination during continuous venovenous haemofiltration and haemodiafiltration in patients with acute renal failure. AB - The elimination of the piperacillin/tazobactam combination was studied in six patients with acute renal failure undergoing either continuous venovenous haemofiltration (CVVH) or continuous venovenous haemodiafiltration (CVVHDF) at 1 L/h and 2 L/h for 12 h. Piperacillin 4 g/tazobactam 0.5 g was given iv on three successive treatment periods and their concentrations in plasma, ultrafiltrate/dialysate and urine were determined for 12 h after each dose. The elimination half-life of piperacillin during CVVH (7.7 +/- 2.3 h; mean +/- s.d.) was significantly longer than during CVVHDF 1 L/h (6.7 +/- 1.9 h) or 2 L/h (6.1 +/- 2.0 h) (P< 0.05). Corresponding values for tazobactam were 13.9 +/- 3.9, 11.6 +/- 3.3 and 9.4 +/- 2.4 h, respectively (P< 0.05). Total piperacillin clearance during CVVH (3.89 +/- 1.23 L/h) was significantly lower than during CVVHDF 1 L/h (5.06 +/- 1.68 L/h) or 2 L/h (5.48 +/- 2.11 L/h) (P< 0.05). The corresponding tazobactam clearance values were 2.42 +/- 0.75, 3.13 +/- 0.66 and 3.75 +/- 1.43 L/h, respectively. The mean 12 h elimination of piperacillin and tazobactam in ultrafiltrate/dialysate was 29% and 37% during CVVH, 42% and 57% during CVVHDF (1 L/h), and 46% and 69% during CVVHDF (2 L/h). We recommend 8 hourly dosing of patients with renal failure on CVVH or CVVHDF with dialysis flow rates of 1 or 2 L/h treated with piperacillin 4 g/tazobactam 0.5 g. PMID- 11733474 TI - An extended interval dosing method for gentamicin in neonates. AB - Traditional gentamicin dosing every 8-24 h depending on age and weight in neonates does not provide the ideal concentration-time profile to both optimize the concentration-dependent killing by aminoglycosides and minimize toxicity. Fifty-three neonates were audited prospectively while receiving gentamicin 2.5 mg/kg every 8-24 h, aiming for peak concentrations (Cmax) of 6-10 mg/L and trough concentrations (Cmin) <2 mg/L. After the first dose, the mean (+/- s.d.) Cmax was 5.5 +/- 0.7 mg/L with sub-therapeutic concentrations (<6 mg/L) in 62% of patients, while the mean Cmin was >2 mg/L in 15% of the neonates. After the third dose the Cmax was 7.5 +/- 1.5 mg/L, with 17% <6 mg/L, whereas the mean Cmin was 2.2 +/- 1.1 mg/L with 49% of values >2 mg/L. An extended interval dosing method (24, 36 and 48 h) for infant weights of 0.75-5 kg was developed by simulation, and audited prospectively in 51 neonates. Prospective analysis of the extended interval dosing method showed a mean Cmax after the first dose of 13.1 +/- 3.6 mg/L, while the mean Cmin was 0.7 +/- 0.6 mg/L. Seventy-eight per cent had Cmax of >10 mg/L after the first dose. The mean area under the concentration versus time curve AUC0-24 was 93 mg*h/L (target = 100 mg*h/L). The extended interval dosing achieved higher Cmax values while ensuring that overall exposure per 24 h was acceptable. Prospective testing of the method demonstrated good predictive ability. PMID- 11733475 TI - Antibiotic failure in the treatment of urinary tract infections in young women. AB - Urinary tract infections (UTIs) are a common problem in young women. The aim of this study was to describe the pattern of antibiotic prescribing to young women presenting with new UTIs and to investigate the proportion who required further treatment if prescribed antibiotics. A secondary aim was to investigate whether the likelihood of treatment failure varied between different antibiotics and, in the case of trimethoprim (the antibiotic most frequently prescribed for UTIs) between prescriptions of different duration. The study included all women aged 15 44 years registered on the UK General Practice Research Database. All diagnoses of UTI or cystitis with an associated prescription for an antibiotic were identified. A further prescription of an antibiotic within 28 days was taken to indicate failure of the initial treatment. Overall, 14% of 75045 newly treated patients with UTI received a second antibiotic within 28 days. Older women, aged 35-44, pregnant patients and those with diabetes were significantly more likely to require further treatment. With trimethoprim as the reference antibiotic, after 28 days patients prescribed amoxicillin were significantly more likely to require a second course of antibiotics. Those prescribed co-trimoxazole were significantly less likely to require further treatment. In each case the difference in failure rate was small and may be of little clinical significance. There was no significant difference between trimethoprim and nitrofurantoin, norfloxacin, ciprofloxacin or the cephalosporins. Three-day prescriptions for trimethoprim appeared as effective as those for 5 or 7 days. This study gives some observational evidence of the effectiveness of antibiotic prescribing in young women with UTIs and shows that between 12% and 16% of patients will return within 28 days for further treatment, irrespective of the antibiotic prescribed initially. PMID- 11733476 TI - Effect of increased inoculum of Salmonella typhi on MIC of azithromycin and resultant growth characteristics. AB - This study was designed to investigate the effect of an increased inoculum on the MIC of azithromycin for Salmonella typhi. Growth curves of nine strains were obtained in cation-adjusted Mueller-Hinton broth containing azithromycin at concentrations of 0-32 mg/L, and comparisons made between inoculation with large inocula, estimated as 10(7) cfu/mL, and with small inocula of c. 10(3) cfu/mL. Turbidity developed only with large inocula after 4-8 h in the presence of 8 or 16 mg/L of azithromycin, thus correlating with microscopic appearance of elongated, curved and widened bacteria. Bacteria that had survived exposure to 16 mg/L for 48 h showed low-grade resistance in comparison with those not exposed to antibiotic. Thus, the mechanism of the inoculum effect was expressed as an enlarging bacterial mass during bacteriostasis, with survival of bacterial populations with low-grade resistance of about two-fold increase in MIC. PMID- 11733477 TI - Influence of the decrease in ciprofloxacin susceptibility and the presence of human serum on the in vitro susceptibility of Streptococcus pneumoniae to five new quinolones. AB - Sixty recent Streptococcus pneumoniae isolates with different susceptibilities to ciprofloxacin (14 with MIC 0.5 mg/L, 10 with MIC 1 mg/L, eight with MIC 2 mg/L, 11 with MIC 4 mg/L and 17 with MIC > or =8 mg/L) were tested against five new quinolones using Todd-Hewitt broth with and without 80% serum. The final inoculum was 5 x 10(5) cfu/mL. Gemifloxacin and clinafloxacin exhibited the lowest MIC90 values and resistance rates (percentage above and defined breakpoint) with and without serum for strains with a ciprofloxacin MIC of > or =4 mg/L. Other quinolones tested were less active against strains with reduced ciprofloxacin susceptibility. The presence of serum did not affect susceptibility to moxifloxacin, but increased the resistance rates to other new quinolones for strains with high ciprofloxacin MICs. PMID- 11733478 TI - Comparative activity of linezolid and other new agents against methicillin resistant Staphylococcus aureus and teicoplanin-intermediate coagulase-negative staphylococci. AB - The activity of linezolid was determined against 225 recently isolated methicillin-resistant Staphylococcus aureus (MRSA) and 20 methicillin-resistant coagulase-negative staphylococci (CoNS) with decreased levels of susceptibility to teicoplanin. Linezolid activity was compared with other new agents (quinupristin-dalfopristin, trovafloxacin, moxifloxacin, levofloxacin and telithromycin) and six other antimicrobials (erythromycin, clindamycin, gentamicin, vancomycin, teicoplanin and rifampicin). The in vitro activity of linezolid was similar to that of vancomycin. Linezolid inhibited all MRSA strains at between 0.1 and 2 mg/L and all CoNS strains tested at between 0.2 and 0.5 mg/L. These results suggest that linezolid would be useful for the treatment of infections involving these organisms. PMID- 11733479 TI - Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and beta-lactam resistance, and to detect common macrolide resistance determinants. AB - Pneumococcal isolates (n = 148) from various countries (mostly from the USA) were tested by a primer set for PCR. Thirty-eight (86.4%) of the 44 penicillin G susceptible isolates (MIC < or = 0.06 mg/L) had unaltered pbps, while six isolates (13.6%) had either one or two alterations in pbps. Of 47 penicillin G resistant strains (MIC > or = 2 mg/L), 41 isolates (87.2%) had all three pbps altered, six isolates (12.8%) had altered pbp1a + 2x. Various combinations of altered pbp were seen in penicillin G-intermediate isolates. Prevalence of macrolide resistance genes mef(A) and erm(B) in isolates was clearly reflected by their MICs. All isolates were positive for lytA. The primers were useful for screening for Streptococcus pneumoniae and beta-lactam resistance, and for detection of common macrolide resistance determinants. PMID- 11733480 TI - In vitro susceptibility of clinical isolates of Zygomycota to amphotericin B, flucytosine, itraconazole and voriconazole. AB - The in vitro susceptibility of 29 clinical isolates of fungi belonging to the division Zygomycota was reviewed. A broth microdilution test was performed by following the NCCLS reference method, with minor modifications. The species of Zygomycota tested were resistant in vitro to flucytosine, itraconazole and voriconazole. Amphotericin B showed the best activity against most of the strains evaluated. PMID- 11733481 TI - Localization of [14C]amoxicillin in rat gastric tissue when administered with lansoprazole and clarithromycin. AB - The gastric mucosal distribution of [14C]amoxicillin when administered to rats with or without lansoprazole and clarithromycin was investigated. After oral administration, the amount found in the gastric mucosa was higher than after iv administration. Co-administration of lansoprazole and clarithromycin had no apparent effect on the distribution pattern of [14C]amoxicillin within the deeper stomach layers. About 50-60% of the radioactivity in the gastric tissue was present in the mucosal layer, irrespective of the route of administration. Microautoradiograms of the gastric mucosa indicated that [14C]amoxicillin was distributed in both the mucous layer and surface epithelial cells following oral administration. [14C]amoxicillin was secreted mainly by surface epithelial cells after iv administration, although only in small quantities. PMID- 11733482 TI - Rifampicin-ofloxacin oral regimen for the treatment of mild to moderate diabetic foot osteomyelitis. AB - Seventeen diabetic patients with moderate to mild foot lesions associated with 20 osteomyelitic bones diagnosed by both bone scan and bone biopsy received rifampicin plus ofloxacin for a median duration of 6 months. Cure was defined as disappearance of all signs and symptoms of infection at the end of the treatment and absence of relapse during follow up. At the end of the treatment period, cure was achieved in 15 patients (88.2%) and was maintained in 13 patients (76.5%) at the end of an average post-treatment follow-up of 22 months. No serious drug related adverse events were recorded. PMID- 11733483 TI - Linezolid levels in pancreatic secretions. PMID- 11733484 TI - Determination of MICs in the routine laboratory. PMID- 11733485 TI - In vitro activity of linezolid against Gram-positive cocci isolated in Poland. PMID- 11733486 TI - Resistance to trimethoprim-sulfamethoxazole and modifications in genes coding for dihydrofolate reductase and dihydropteroate synthase in European Streptococcus pneumoniae isolates. PMID- 11733487 TI - Identification of reduced fluoroquinolone susceptibility in Escherichia coli: a herald for emerging resistance. PMID- 11733488 TI - Characterization of the translational attenuator of 20 methicillin-resistant, quinupristin/dalfopristin-resistant Staphylococcus aureus isolates with reduced susceptibility to glycopeptides. PMID- 11733489 TI - Glycopeptide resistance in an epidemic strain of methicillin-resistant Staphylococcus aureus. PMID- 11733490 TI - Peroxisome proliferator-activated receptor gamma coactivator 1beta (PGC-1beta ), a novel PGC-1-related transcription coactivator associated with host cell factor. AB - Peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1) plays a critical role in regulating multiple aspects of energy metabolism, including adaptive thermogenesis, mitochondrial biogenesis, and fatty acid beta-oxidation. Recently, this coactivator of nuclear receptors/transcription factors has been shown to control hepatic gluconeogenesis, an important component of the pathogenesis of both type-1 and type-2 diabetes. We described here the cloning of a novel bona fide homologue of PGC-1, PGC-1beta (PGC-1 was renamed as PGC 1alpha), first identified through searches of new data base entries. Despite the fact that PGC-1alpha and -1beta share similar tissue distributions with highest levels of expression in brown fat and heart, their mRNAs are differentially regulated in the brown adipose tissue upon cold exposure and during brown fat cell differentiation. Like PGC-1alpha, PGC-1beta mRNA levels are increased significantly in the liver during fasting, suggesting a possible role for this factor in the regulation of hepatic gluconeogenesis and/or fatty acid oxidation. Consistent with this, PGC-1beta was shown to physically interact and potently coactivate hepatic nuclear factor 4 and peroxisome proliferator-activated receptor alpha, nuclear receptors that are essential for hepatic adaptation to fasting. Finally, using sequence comparisons between PGC-1alpha and -1beta, we have identified a conserved amino acid motif that serves as a docking site for host cell factor, a cellular protein implicated in cell cycle regulation and viral infection. HCF is shown to bind to both PGC-1alpha and -1beta and augment their transcriptional activity. PMID- 11733491 TI - Characterization of the H-kininogen-binding site on factor XI: a comparison of factor XI and plasma prekallikrein. AB - Factor XI (FXI), the zymogen of the blood coagulation protease FXIa, and the structurally homologous protein plasma prekallikrein circulate in plasma in noncovalent complexes with H-kininogen (HK). HK binds to the heavy chains of FXI and of prekallikrein. Each chain contains four apple domains (F1-F4 for FXI and P1-P4 for prekallikrein). Previous studies indicated that the HK-binding site on FXI is located in F1, whereas the major HK-binding site on prekallikrein is in P2. To determine the contribution of each FXI apple domain to HK-FXI complex formation, we examined binding of recombinant single apple domain-tissue plasminogen activator fusion proteins to HK. The order of affinity from highest to lowest is F2 F4 > F1 F3. Monoclonal antibodies against F2 are superior to F4 or F1 antibodies as inhibitors of HK binding to FXI. Antibody alphaP2, raised against prekallikrein, cross-reacts with FXI F2 and inhibits FXI-HK binding with an IC(50) of 8 nm. HK binding to a platelet-specific FXI variant lacking the N terminal half of F2 is reduced > 5-fold compared with full-length FXI. A chimeric FXI molecule in which F2 is replaced by P2 is cleaved within P2 during activation by factor XIIa, resulting in greatly reduced HK binding capacity. In contrast, wild-type FXI is not cleaved within F2, and its binding capacity for HK is unaffected by factor XIIa. Our data show that HK binding to FXI involves multiple apple domains, with F2 being most important. The findings demonstrate a similarity in mechanism for FXI and prekallikrein binding to HK. PMID- 11733492 TI - Crystal structure of the extracellular domain of mouse RANK ligand at 2.2-A resolution. AB - Bone remodeling involves the resorption of bone by osteoclasts and the synthesis of bone matrix by osteoblasts. Receptor activator of NF-kappa B ligand (RANKL, also known as ODF and OPGL), a member of the tumor necrosis factor (TNF) family, triggers osteoclastogenesis by forming a complex with its receptor, RANK. We have determined the crystal structure of the extracellular domain of mouse RANKL at 2.2-A resolution. The structure reveals that the RANKL extracellular domain is trimeric, which was also shown by analytical ultracentrifugation, and each subunit has a beta-strand jellyroll topology like the other members of the TNF family. A comparison of RANKL with TNF beta and TNF-related apoptosis-inducing ligand (TRAIL), whose structures were determined to be in the complex form with their respective receptor, reveals conserved and specific features of RANKL in the TNF superfamily and suggests the presence of key residues of RANKL for receptor binding. PMID- 11733493 TI - Mitochondrial protein import: molecular basis of the ATP-dependent interaction of MtHsp70 with Tim44. AB - Protein translocation across the mitochondrial inner membrane is driven by cycles of binding and release of mitochondrial heat shock protein 70 (mtHsp70) in the matrix. The peripheral inner membrane protein, Tim44, recruits mtHsp70 in an ATP dependent manner to the import sites. We show that DnaK, the closely related Hsp70 of Escherichia coli, when targeted to the matrix of yeast mitochondria, interacts in a specific manner with Tim44. The interaction is, however, not regulated by ATP, and DnaK cannot support protein translocation. We used truncated mtHsp70s and chimeric proteins consisting of segments of mtHsp70 and DnaK to analyze which portions of mtHsp70 bind and functionally interact with Tim44. We show that Tim44 interacts with the beta-stranded core of the peptide binding domain of mtHsp70 and of DnaK. The alpha-helices A and B of the peptide binding domain of mtHsp70 are required to transmit the nucleotide state of the ATPase domain to the peptide binding domain. Tim44, by interacting in this way with the peptide binding domain, is proposed to coordinate the binding of mtHsp70 to the incoming preprotein and the subsequent release of the mtHsp70-preprotein complex from the TIM23 complex, the translocase of the inner membrane. PMID- 11733494 TI - Activation of large conductance sodium channels upon expression of amiloride sensitive sodium channel in Sf9 insect cells. AB - The amiloride-sensitive epithelial sodium channels (ENaC) mediate Na(+) reabsorption in epithelial tissues including distal nephron, colon, lung, and secretory glands and plays a critical role in pathophysiology of hypertension and cystic fibrosis. The ENaC is a multimeric protein composed of alpha-ENaC, beta ENaC, and gamma-ENaC subunits. To study the biochemical properties of the channel, the subunit cDNAs of rat colon ENaC (rENaC) were subcloned into baculoviruses, and the corresponding proteins were expressed in Sf9 insect cells. The functional characteristics of the expressed rENaC were studied in planar lipid bilayers. The results show that expression of alpha-rENaC and alphabetagamma-rENaC in Sf9 insect cells results in the generation of cation selective large conductance channels. Although the large conductance channels observed in the alpha-rENaC-containing membranes were unaffected by amiloride, the large conductance channels found in alphabetagamma-rENaC complex-containing membranes exhibited voltage-dependent flickering in the presence of micromolar amiloride. Possible implications of these observations are discussed. PMID- 11733495 TI - Interferon-gamma-mediated activation and ubiquitin-proteasome-dependent degradation of PPARgamma in adipocytes. AB - Interferon-gamma (IFNgamma) treatment of adipocytes results in a down-regulation of the peroxisome proliferator-activated receptor gamma (PPARgamma). The decrease in PPARgamma expression is mediated by inhibition of PPARgamma synthesis and increased degradation of PPARgamma. In this study, we demonstrate that both PPARgamma1 and PPARgamma2 are targeted to the proteasome under basal conditions and that PPARgamma1 is more labile than PPARgamma2. The IFNgamma-induced increase in PPARgamma turnover is blocked by proteasome inhibition and is accompanied by an increase in PPARgamma-polyubiquitin conjugates. In addition, IFNgamma treatment results in the transcriptional activation of PPARgamma. Similar to ligand-dependent activation of PPARgamma, IFNgamma-induced activation was greater in the phosphorylation-deficient S112A form of PPARgamma when compared with wild type PPARgamma. Moreover, the inhibition of ERKs 1 and 2 with a MEK inhibitor, U1026, lead to an inhibition in the decay of PPARgamma proteins, indicating that serine phosphorylation influences the degradation of PPARgamma in fat cells. Our results also demonstrate that the proteasome-dependent degradation of PPARgamma does not require nuclear export. Taken together, these results indicate that PPARgamma is targeted to the ubiquitin-proteasome pathway for degradation under basal conditions and that IFNgamma leads to an increased targeting of PPARgamma to the ubiquitin-proteasome system in a process that is affected by ERK-regulated serine phosphorylation of PPARgamma proteins. PMID- 11733496 TI - Regulation of cytosolic phospholipase A2 activity in macrophages stimulated with receptor-recognized forms of alpha 2-macroglobulin: role in mitogenesis and cell proliferation. AB - Macrophages exposed to receptor-recognized forms of alpha(2)-macroglobulin (alpha(2)M*) demonstrate increased DNA synthesis and cell division. In the current study, we have probed the role of cytosolic phospholipase A(2) (cPLA(2)) activity in the cellular response to alpha(2)M*. Ligation of the alpha(2)M* signaling receptor by alpha(2)M*, or its receptor binding fragment, increased cPLA(2) activity 2-3-fold in a concentration and time-dependent manner. This activation required a pertussis toxin-insensitive G protein. Cellular binding of alpha(2)M* also induced transient translocation of cPLA(2) activity to nuclei and membrane fractions. Inhibition of protein kinase C activity or chelation of Ca(2+) inhibited alpha(2)M*-induced increased cPLA(2) activity. Binding of alpha(2)M* to macrophages, moreover, increased phosphorylation of MEK 1/2, ERK 1/2, p38 MAPK, and JNK. Incubation of macrophages with inhibitors of MEK 1/2 or p38 MAPK before stimulation with alpha(2)M* profoundly decreased phosphorylation of MAPKs, blocking cPLA(2) activation. alpha(2)M*-induced increase in [(3)H]thymidine uptake and cell proliferation was completely abolished if activation of cPLA(2) was prevented. The response of macrophages to alpha(2)M* requires transcription factors nuclear factor kappaB, and cAMP-responsive element binding protein as well as expression of the proto-oncogenes c-fos and c-myc. These studies indicate that the activation of cPLA(2) plays a crucial role in alpha(2)M*-induced mitogenesis and cell proliferation. PMID- 11733497 TI - A novel leucine zipper targets AKAP15 and cyclic AMP-dependent protein kinase to the C terminus of the skeletal muscle Ca2+ channel and modulates its function. AB - In skeletal muscle, voltage-dependent potentiation of L-type Ca(2+) channel (Ca(V)1.1) activity requires phosphorylation by cyclic AMP-dependent protein kinase (PKA) anchored via an A kinase-anchoring protein (AKAP15). However, the mechanism by which AKAP15 targets PKA to L-type Ca(2+) channels has not been elucidated. Here we report that AKAP15 directly interacts with the C-terminal domain of the alpha(1) subunit of Ca(V)1.1 via a leucine zipper (LZ) motif. Disruption of the LZ interaction effectively inhibits voltage-dependent potentiation of L-type Ca(2+) channels in skeletal muscle cells. Our results reveal a novel mechanism whereby anchoring of PKA to Ca(2+) channels via LZ interactions ensures rapid and efficient phosphorylation of Ca(2+) channels in response to local signals such as cAMP and depolarization. PMID- 11733498 TI - Interaction between active Pak1 and Raf-1 is necessary for phosphorylation and activation of Raf-1. AB - Activation of Raf-1 is a complex process in which phosphorylation of Ser(338) Tyr(341) is a critical step. Previous studies have shown that Pak1/2 is implicated in both Ras-dependent and -independent activation of Raf-1 by phosphorylating Raf Ser(338). The present study explores the structural basis of Raf-1 phosphorylation by Pak1. We found that Pak directly associates with Raf-1 under both physiological and overexpressed conditions. The association is greatly stimulated by 4beta-12-O-tetradecanoylphorbol-13-acetate and nocodazole and by expression of the active mutants of Rac and Ras. The active forms of Pak generated by mutation of Thr(423) to Glu or truncation of the amino-terminal moiety exhibit a greater binding to Raf than the wild type, whereas the kinase dead mutant Pak barely binds Raf. The extent of binding to Raf-1 is correlated with the ability of Pak to phosphorylate Raf and induce mitogen-activated protein kinase activation. Furthermore, the Raf-1 binding site is defined to the carboxyl terminus of the Pak catalytic domain. In addition, our results suggest that the amino-terminal regulatory region of Raf inhibits the interaction. Taken together, the results indicate that the interaction depends on the active conformations of Pak and Raf. They also argue that Pak1 is a physiological candidate for phosphorylation of Raf Ser(338) during the course of Raf activation. PMID- 11733499 TI - Accurate chromatin organization of the mouse mammary tumor virus promoter determines the nature of the synergism between transcription factors. AB - The mechanism underlying the synergism between transcription factors in eukaryotic gene expression is not fully understood. In minichromosomes assembled in vitro the synergism between steroid hormone receptors (SHRs) and nuclear factor 1 (NF1) on the mouse mammary tumor virus (MMTV) promoter does not require the proline-rich transactivation domain (PRD) of NF1. Here we show that similar results are obtained in yeast. In contrast, replacing the native hormone responsive elements (HREs) by a single HRE results in a more accessible chromatin and makes the synergism with SHR dependent on the PRD of NF1. Following hormone induction, in addition to glucocorticoid receptor, the DNA binding domain of NF1 is needed and sufficient for establishing an open chromatin conformation on the wild type MMTV promoter. Thus, NF1 acts as a classical transcription factor in a relaxed chromatin context, whereas in the context of the wild type chromatin DNA binding of NF1 is sufficient to cooperate with SHRs by stabilizing an open chromatin conformation. PMID- 11733500 TI - Enterocyte expression of the eotaxin and interleukin-5 transgenes induces compartmentalized dysregulation of eosinophil trafficking. AB - Eosinophils accumulate in the gastrointestinal tract in a number of medical disorders, but the mechanisms involved are largely unknown. To understand the significance of cytokine expression by enterocytes, enterocyte transgenic mice that overexpressed the eosinophil-selective cytokines eotaxin and interleukin (IL)-5 were generated. Transgenic mice, generated by utilizing the rat intestinal fatty acid-binding protein promoter (Fabpi), overexpressed the mRNA for these cytokines in the small intestine. Overexpression of IL-5 resulted in marked increases of eosinophils in the bone marrow and blood, whereas eotaxin overexpression resulted in similar levels compared with nontransgenic control mice. In contrast, both IL-5 and eotaxin transgenic mice had significant accumulation of eosinophils in the gastrointestinal mucosa compared with control mice. Eotaxin-induced gastrointestinal eosinophilia was substantially higher than that induced by IL-5 and was especially prominent within the lamina propria of the villi. Interestingly, genetic rescue of eotaxin deficiency (by transgenic overexpression of eotaxin in eotaxin gene-targeted mice) resulted in significant restoration of gastrointestinal eosinophil levels. Finally, the intestinal eosinophilia induced by the eotaxin transgene was beta(7) integrin-dependent. Taken together, these results demonstrate that expression of eotaxin and IL-5 in intestinal epithelium induces compartmentalized dysregulation of eosinophil trafficking and the important role of the beta(7) integrin in gastrointestinal allergic responses. PMID- 11733501 TI - Stereo-specific substrate recognition by phosphatidylinositol phosphate kinases is swapped by changing a single amino acid residue. AB - Type I and type II phosphatidylinositol phosphate (PIP) kinases generate the lipid second messenger phosphatidylinositol (PtdIns) 4,5-bisphosphate and thus play fundamental roles in the regulation of many cellular processes. Although the two kinase families are highly homologous, they phosphorylate distinct substrates and are functionally non-redundant. Type I PIP kinases phosphorylate PtdIns 4 phosphate at the D-5 hydroxyl group and are consequently PtdIns 4-phosphate 5 kinases. By contrast, type II PIP kinases are PtdIns 5-phosphate 4-kinases that phosphorylate PtdIns 5-phosphate at the D-4 position. Type I PIP kinases, in addition, also phosphorylate other phosphoinositides in vitro and in vivo and thus have the potential to generate multiple lipid second messengers. To understand how these enzymes differentiate between stereoisomeric substrates, we used a site-directed mutagenesis approach. We show that a single amino acid substitution in the activation loop, A381E in IIbeta and the corresponding mutation E362A in Ibeta, is sufficient to swap substrate specificity between these PIP kinases. In addition to its role in substrate specificity, the type I activation loop is also key in subcellular targeting. The Ibeta(E362A) mutant and other mutants with reduced PtdIns 4-phosphate binding affinity were largely cytosolic when expressed in mammalian cells in contrast to wild-type Ibeta which targets to the plasma membrane. These results clearly establish the role of the activation loop in determining both signaling specificity and plasma membrane targeting of type I PIP kinases. PMID- 11733502 TI - Ku represses the HIV-1 transcription: identification of a putative Ku binding site homologous to the mouse mammary tumor virus NRE1 sequence in the HIV-1 long terminal repeat. AB - Ku has been implicated in nuclear processes, including DNA break repair, transcription, V(D)J recombination, and telomere maintenance. Its mode of action involves two distinct mechanisms: one in which a nonspecific binding occurs to DNA ends and a second that involves a specific binding to negative regulatory elements involved in transcription repression. Such elements were identified in mouse mammary tumor virus and human T cell leukemia virus retroviruses. The purpose of this study was to investigate a role for Ku in the regulation of human immunodeficiency virus (HIV)-1 transcription. First, HIV-1 LTR activity was studied in CHO-K1 cells and in CH0-derived xrs-6 cells, which are devoid of Ku80. LTR-driven expression of a reporter gene was significantly increased in xrs-6 cells. This enhancement was suppressed after re-expression of Ku80. Second, transcription of HIV-1 was followed in U1 human cells that were depleted in Ku by using a Ku80 antisense RNA. Ku depletion led to a increase of both HIV-1 mRNA synthesis and viral production compared with the parent cells. These results demonstrate that Ku acts as a transcriptional repressor of HIV-1 expression. Finally, a putative Ku-specific binding site was identified within the negative regulatory region of the HIV-1 long terminal repeat, which may account for this repression of transcription. PMID- 11733503 TI - Effects of B-Myb on gene transcription: phosphorylation-dependent activity ans acetylation by p300. AB - The transcription factor B-Myb is a cell-cycle regulated phosphoprotein involved in cell cycle progression through the transcriptional regulation of many genes. In this study, we show that the promoter of the fibroblast growth factor-4 (FGF 4) gene is strongly activated by B-Myb in HeLa cells and it can serve as a novel diagnostic tool for assessing B-Myb activity. Specifically, B-Myb deletion mutants were examined and domains of B-Myb required for activation of the FGF-4 promoter were identified. Using phosphorylation-deficient mutant forms of B-Myb, we also show that phosphorylation is essential for B-Myb activity. Moreover, a mutant form of B-Myb, which lacks all identified phosphorylation sites and which has little activity, can function as a dominant-negative and suppress wild-type B Myb activity. Acetylation is another post-translational modification known to affect the activity of other Myb family members. We show that B-Myb is acetylated by the co-activator p300. We also show that the bromo and histone acetyltransferase domains of p300 are sufficient to interact with and acetylate B Myb. These data indicate that phosphorylation of B-Myb is an essential modification for activity and that acetylation of B-Myb may play a role in B-Myb activity. PMID- 11733504 TI - Interaction with the small subunit of geranyl diphosphate synthase modifies the chain length specificity of geranylgeranyl diphosphate synthase to produce geranyl diphosphate. AB - Geranyl diphosphate synthase belongs to a subgroup of prenyltransferases, including farnesyl diphosphate synthase and geranylgeranyl diphosphate synthase, that catalyzes the specific formation, from C(5) units, of the respective C(10), C(15), and C(20) precursors of monoterpenes, sesquiterpenes, and diterpenes. Unlike farnesyl diphosphate synthase and geranylgeranyl diphosphate synthase, which are homodimers, geranyl diphosphate synthase from Mentha is a heterotetramer in which the large subunit shares functional motifs and a high level of amino acid sequence identity (56-75%) with geranylgeranyl diphosphate synthases of plant origin. The small subunit, however, shares little sequence identity with other isoprenyl diphosphate synthases; yet it is absolutely required for geranyl diphosphate synthase catalysis. Coexpression in Escherichia coli of the Mentha geranyl diphosphate synthase small subunit with the phylogenetically distant geranylgeranyl diphosphate synthases from Taxus canadensis and Abies grandis yielded a functional hybrid heterodimer that generated geranyl diphosphate as product in each case. These results indicate that the geranyl diphosphate synthase small subunit is capable of modifying the chain length specificity of geranylgeranyl diphosphate synthase (but not, apparently, farnesyl diphosphate synthase) to favor the production of C(10) chains. Comparison of the kinetic behavior of the parent prenyltransferases with that of the hybrid enzyme revealed that the hybrid possesses characteristics of both geranyl diphosphate synthase and geranylgeranyl diphosphate synthase. PMID- 11733505 TI - Chlorination of bacterial and neutrophil proteins during phagocytosis and killing of Staphylococcus aureus. AB - Myeloperoxidase is proposed to play a central role in bacterial killing by generating hypochlorous acid within neutrophil phagosomes. However, it has yet to be demonstrated that these inflammatory cells target hypochlorous acid against bacteria inside phagosomes. In this investigation, we treated Staphylococcus aureus with varying concentrations of reagent hypochlorous acid and found that even at sublethal doses, it converted some tyrosine residues in their proteins to 3-chlorotyrosine and 3,5-dichlorotyrosine. To determine whether or not ingested bacteria were exposed to hypochlorous acid in neutrophil phagosomes, we labeled their proteins with [(13)C(6)]tyrosine and used gas chromatography with mass spectrometry to identify the corresponding chlorinated isotopes after the bacteria had been phagocytosed. Chlorinated tyrosines were detected in bacterial proteins 5 min after phagocytosis and reached levels of approximately 2.5/1000 mol of tyrosine at 60 min. Inhibitor studies revealed that chlorination was dependent on myeloperoxidase. Chlorinated neutrophil proteins were also detected and accounted for 94% of total chlorinated tyrosine residues formed during phagocytosis. We conclude that hypochlorous acid is a major intracellular product of the respiratory burst. Although some reacts with the bacteria, most reacts with neutrophil components. PMID- 11733506 TI - A novel binding protein composed of homophilic tetramer exhibits unique properties for the small GTPase Rab5. AB - The small GTPase Rab family, which cycles between GTP-bound active and GDP-bound inactive states, plays an important role in membrane trafficking. Among them, Rab5 is involved in early endocytic pathway, and several Rab5-binding proteins have been identified as regulators or effectors to coordinate the docking and fusion processes of endocytic vesicles. We describe a novel binding protein exhibiting unique biochemical properties for Rab5. The Rab5-binding protein enhances GDP-GTP exchange reaction on Rab5 but preferentially interacts with its GTP-bound form. Gel filtration and immunoprecipitation analyses indicate that the Rab5-binding protein functions as a tetramer composed of anti-parallel linkage of two parallel dimers. These results suggest that the newly identified protein may function as an upstream activator and/or downstream effector for Rab5 in endocytic pathway. Possible roles of the quaternary structure have been discussed in terms of the Rab5-mediated signaling. PMID- 11733507 TI - Isolated plant mitochondria import chloroplast precursor proteins in vitro with the same efficiency as chloroplasts. AB - Most chloroplast and mitochondrial proteins are synthesized with N-terminal presequences that direct their import into the appropriate organelle. In this report we have analyzed the specificity of standard in vitro assays for import into isolated pea chloroplasts and mitochondria. We find that chloroplast protein import is highly specific because mitochondrial proteins are not imported to any detectable levels. Surprisingly, however, pea mitochondria import a range of chloroplast protein precursors with the same efficiency as chloroplasts, including those of plastocyanin, the 33-kDa photosystem II protein, Hcf136, and coproporphyrinogen III oxidase. These import reactions are dependent on the Deltaphi across the inner mitochondrial membrane, and furthermore, marker enzyme assays and Western blotting studies exclude any import by contaminating chloroplasts in the preparation. The pea mitochondria specifically recognize information in the chloroplast-targeting presequences, because they also import a fusion comprising the presequence of coproporphyrinogen III oxidase linked to green fluorescent protein. However, the same construct is targeted exclusively into chloroplasts in vivo indicating that the in vitro mitochondrial import reactions are unphysiological, possibly because essential specificity factors are absent in these assays. Finally, we show that disruption of potential amphipathic helices in one presequence does not block import into pea mitochondria, indicating that other features are recognized. PMID- 11733508 TI - The conserved active site asparagine in class I ribonucleotide reductase is essential for catalysis. AB - The active site residue Asn-437 in protein R1 of the Escherichia coli ribonucleotide reductase makes a hydrogen bond to the 2'-OH group of the substrate. To elucidate its role(s) during catalysis, Asn-437 was engineered by site-directed mutagenesis to several other side chains (Ala, Ser, Asp, Gln). All mutant proteins were incapable of enzymatic turnover but promoted rapid protein R2 tyrosyl radical decay in the presence of the k(cat) inhibitor 2'-azido-2' deoxy-CDP with similar decay rate constants as the wild-type R1. These results show that all Asn-437 mutants can perform 3'-H abstraction, the first substrate related step in the reaction mechanism. The most interesting observation was that three of the mutant proteins (N437A/S/D) behaved as suicidal enzymes by catalyzing a rapid tyrosyl radical decay also in reaction mixtures containing the natural substrate CDP. The suicidal CDP-dependent reaction was interpreted to suggest elimination of the substrate's protonated 2'-OH group in the form of water, a step that has been proposed to drive the 3'-H abstraction step. A furanone-related chromophore was formed in the N437D reaction, which is indicative of stalling of the reaction mechanism at the reduction step. We conclude that Asn-437 is essential for catalysis but not for 3'-H abstraction. We propose that the suicidal N437A, N437S, and N437D mutants can also catalyze the water elimination step, whereas the inert N437Q mutant cannot. Our results suggest that Asn-437, apart from hydrogen bonding to the substrate, also participates in the reduction steps of catalysis by class I ribonucleotide reductase. PMID- 11733509 TI - Formation of functional heterodimers between the TASK-1 and TASK-3 two-pore domain potassium channel subunits. AB - The potassium channels in the two-pore domain family are widely expressed and regulate the excitability of neurons and other excitable cells. These channels have been shown to function as dimers, but heteromerization between the various channel subunits has not yet been reported. Here we demonstrate that two members of the TASK subfamily of potassium channels, TASK-1 and TASK-3, can form functional heterodimers when expressed in Xenopus laevis oocytes. To recognize the two TASK channel types, we took advantage of the higher sensitivity of TASK-1 over TASK-3 to physiological pH changes and the discriminating sensitivity of TASK-3 to the cationic dye ruthenium red. These features were clearly observed when the channels were expressed individually. However, when TASK-1 and TASK-3 were expressed together, the resulting current showed intermediate pH sensitivity and ruthenium red insensitivity (characteristic of TASK-1), indicating the formation of TASK-1/TASK-3 heterodimers. Expression of a tandem construct in which TASK-3 and TASK-1 were linked together yielded currents with features very similar to those observed when coexpressing the two channels. The tandem construct also responded to AT(1a) angiotensin II receptor stimulation with an inhibition that was weaker than the inhibition of homodimeric TASK-1 and greater than that shown by TASK-3. Expression of epitope-tagged channels in mammalian cells showed their primary presence in the plasma membrane consistent with their function in this location. Heteromerization of two-pore domain potassium channels may provide a greater functional diversity and additional means by which they can be regulated in their native tissues. PMID- 11733510 TI - Functional expression of heteromeric calcitonin gene-related peptide and adrenomedullin receptors in yeast. AB - The ability of G protein-coupled receptors (GPCRs) to form homo- and heteromeric complexes has important implications for the regulation of cellular events. A notable example of heteromer formation is the interaction of the calcitonin receptor-like receptor (CRLR) with different members of the receptor activity modifying protein (RAMP) family, which results in the formation of two different receptors, a calcitonin gene-related peptide (CGRP) receptor and an adrenomedullin receptor. To analyze the role of RAMPs in determining ligand specificity, we have co-expressed CRLR and RAMP proteins in the yeast Saccharomyces cerevisiae, which provides a null system to study the function of mammalian receptors. Co-expression of RAMP1 and CRLR reconstituted a CGRP receptor that was able to activate the pheromone-signaling pathway with pharmacological properties similar to those observed previously in mammalian cells. Co-expression of CRLR with RAMP2 or RAMP3 resulted in a response with the pharmacological properties of an adrenomedullin receptor. These data indicate that RAMPs are necessary and sufficient to determine ligand specificity of CRLR. Contrary to observations in mammalian cells, the glycosylation of CRLR was not affected by the presence of RAMPs in yeast, indicating that glycosylation of CRLR is not the prime determinant of ligand specificity. The first functional reconstitution of a heteromeric seven transmembrane receptor in yeast suggests this organism as a useful research tool to study the molecular nature of other heteromeric receptors. PMID- 11733511 TI - Bile salt-stimulated carboxyl ester lipase influences lipoprotein assembly and secretion in intestine: a process mediated via ceramide hydrolysis. AB - Bile salt-stimulated carboxyl ester lipase (CEL), also called cholesterol esterase, is one of the major proteins secreted by the pancreas. The physiological role of CEL was originally thought to be its mediation of dietary cholesterol absorption. However, recent studies showed no difference between wild type and CEL knockout mice in the total amount of cholesterol absorbed in a single meal. The current study tests the hypothesis that CEL in the intestinal lumen may influence the type of lipoproteins produced. A lipid emulsion containing 4 mm phospholipid, 13.33 mm [(3)H]triolein, and 2.6 mm [(14)C]cholesterol in 19 mm taurocholate was infused into the duodenum of lymph fistula CEL(+/+) and CEL(-/-) mice at a rate of 0.3 ml/h. Results showed no difference between CEL(+/+) and CEL(-/-) mice in the rate of cholesterol and triglyceride transport from the intestinal lumen to the lymph. However, CEL(-/-) mice produced predominantly smaller lipoproteins, whereas the CEL(+/+) mice produced primarily large chylomicrons and very low density lipoprotein. The proximal intestine of CEL(-/-) mice was also found to possess significantly less ceramide hydrolytic activity than that present in CEL(+/+) mice. By using Caco2 cells grown on Transwell membranes as a model, sphingomyelinase treatment inhibited the secretion of larger chylomicron-like lipoproteins without affecting total cholesterol secretion. In contrast, the addition of CEL to the apical medium increased the amount of large lipoproteins produced and alleviated the inhibition induced by sphingomyelinase. Taken together, this study identified a novel and physiologically significant role for CEL, namely the promotion of large chylomicron production in the intestine. The mechanism appears to be mediated through CEL hydrolysis of ceramide generated during the lipid absorption process. PMID- 11733512 TI - GATA-6 and thyroid transcription factor-1 directly interact and regulate surfactant protein-C gene expression. AB - GATA-6, a member of the GATA family of zinc finger domain containing transcription factors, is expressed in endodermally derived tissues including the lung, where GATA-6 influences the transcription of target genes, TTF-1, and surfactant proteins. Whereas GATA-6 did not directly alter expression of sp-C constructs in HeLa cells, GATA-6 synergistically activated sp-C gene transcription when co-expressed with TTF-1, supporting the concept that GATA-6 and TTF-1 might directly interact to influence target gene expression. GST-GATA-6 directly co-precipitated with TTF-1 after in vitro translation and directly interacted with the TTF-1-binding element in the sp-C promoter. Binding of TTF-1 to GATA-6 required the homeodomain of TTF-1, but optimal interactions with GATA-6 required the homeodomain and either carboxyl- or amino-terminal domains of TTF-1. Interactions between TTF-1 and GATA-6 required the amino-terminal and zinc finger domains of GATA-6. Although GATA-4 also interacted with TTF-1 in two-hybrid assays, GATA-4 was not as active as GATA-6 in the activation of the sp-C promoter with TTF-1. Deletion and substitution mutations between GATA-4 and GATA-6 demonstrated that the carboxyl-terminal zinc finger domain of GATA-6 contributed to its synergistic activation of the sp-C promoter with TTF-1. GATA-6 influenced the activity of the sp-C promoter, binding and acting synergistically with TTF-1. PMID- 11733513 TI - Inhibition of T cell antigen receptor signaling by VHR-related MKPX (VHX), a new dual specificity phosphatase related to VH1 related (VHR). AB - A cDNA encoding a novel, human, dual-specific protein phosphatase was identified in the Incyte data base. The open reading frame predicted a protein of 184 amino acids related to the Vaccinia virus VH1 and human VH1-related (VHR) phosphatases. Expression VHR-related MKPX (VHX) was highest in thymus, but also detectable in monocytes and lymphocytes. A VHX-specific antiserum detected a protein with an apparent molecular mass of 19 kDa in many cells, including T lymphocytes and monocytes. VHX expression was not induced by T cell activation, but decreased somewhat at later time points. In vitro, VHX dephosphorylated the Erk2 mitogen activated protein kinase with faster kinetics than did VHR, which is thought to be specific for Erk1 and 2. When expressed in Jurkat T cells, VHX had the capacity to suppress T cell antigen receptor-induced activation of Erk2 and of an NFAT/AP-1 luciferase reporter, but not an NF-kappaB reporter. Thus, VHX is a new member of the VH1/VHR group of small dual-specific phosphatases that act in mitogen-activated protein kinase signaling pathways. PMID- 11733514 TI - Collaboration of JNKs and ERKs in nerve growth factor regulation of the neurofilament light chain promoter in PC12 cells. AB - Nerve growth factor (NGF) induces transcription-dependent neural differentiation of PC12 cells, and the ERK family of MAPKs has been implicated as the dominant signal pathway that mediates this response. We employed a neurofilament light chain (NFLC) promoter-luciferase (NFLC-Luc) reporter to define the role of the ERKs as well as additional MAPK pathways in NGF induction of this neural specific gene. Constitutive active forms of c-Raf-1, MEKK1 and MKK6, proximal regulators of the ERKs, JNKs, and p38 MAPKs, respectively, all stimulated NFLC-Luc activity. NFLC-Luc activity stimulated by NGF, however, was partially (approximately 50%) inhibited by the MEK inhibitor, PD098059, or by co-transfection of kinase inactive MEK1 but not by the p38 MAPK inhibitor, SB203580, indicating a role for the ERKs, but not the p38 MAPKs, in NGF regulation of the NFLC promoter. Importantly, a gain-of-function MKK7-JNK3 fusion protein stimulated NFLC-Luc and synergized with gain-of-function c-Raf-1 to activate the NFLC promoter. In addition, transfection of kinase-inactive forms of MEK1 and MKK7 produced an additive inhibition of NGF-stimulated NFLC-Luc relative to either inhibitor alone. These findings indicate that the ERK and JNK pathways collaborate downstream of the NGF receptor for regulation of the NFLC promoter. Truncation analysis and electromobility shift assays established the requirement for a cAMP response element/activating transcription factor-like site in the NFLC promoter that minimally interacts with constitutively expressed cAMP-response element binding protein and JunD as well as c-Jun which is induced by NGF in an ERK dependent manner. Cumulatively, these findings indicate that the ERK pathway requires collaboration with the JNK pathway for maximal activation of the NFLC gene in PC12 cells through the integrated control of c-Jun function. PMID- 11733516 TI - CCAAT/enhancer-binding protein beta plays a regulatory role in differentiation and apoptosis of neuroblastoma cells. AB - The C/EBPbeta (CCAAT/enhancer-binding protein beta) is a transcription factor that belongs to basic region-leucine zipper class DNA-binding proteins. There is a significant body of evidence that suggests that this protein plays a central role in adipocytic and eosinophilic differentiation. However, there is no information available regarding the role of this transcription factor in the development of mammalian neuronal tissues. In this study, we have examined the effect of C/EBPbeta overexpression on the differentiation and survival of mouse Neuro2A cells. We found that C/EBPbeta induces neuronal differentiation and that this process is inhibited by transfection with the C/EBP homologous protein 10 (CHOP), strongly suggesting that the extension of neurites is indeed due to the C/EBPbeta transcriptional activity. As it has been suggested in adipocyte differentiation, here we show that C/EBPbeta induces the expression of the endogenous C/EBPalpha gene and that this protein by itself is also able to induce a differentiated phenotype in Neuro2A cells. Neuronal differentiation induced by C/EBPbeta requires activation of the phosphatidylinositol 3-kinase signaling pathway, whereas inhibition of the mitogen-activated protein kinase signaling does not have any effect. In addition, we show that C/EBPbeta is expressed in the brain of neonatal rats, suggesting that this protein could play an important role in neuronal maturation. Finally, cell death was also induced by C/EBPbeta through activation of the p53 protein and the cdk inhibitor p21. PMID- 11733515 TI - Regulation of Fas expression by STAT3 and c-Jun is mediated by phosphatidylinositol 3-kinase-AKT signaling. AB - Cooperation between STAT3 and c-Jun results in suppression of Fas Receptor (FasR) transcription, which is often seen in advanced human tumors. To identify requirements for STAT3-Jun cooperation, we elucidated the role of protein kinases that affect both transcription factors. The phosphatidylinositol 3-kinase (PI3K) AKT signaling pathway was found capable of down-regulating both STAT3- and c-Jun dependent transcription, resulting in derepression of FasR transcription. Conversely, inhibition of PI3K-AKT signaling via the specific pharmacological inhibitor LY294002 up-regulated AP1/Jun- and STAT-dependent transcriptional activities, resulting in suppression of the FasR promoter activities and decreased FasR surface expression. PI3K-AKT's ability to affect FasR transcription was not observed in c-jun null fibroblasts, suggesting that c-Jun is required for PI3K/AKT-mediated regulation of FasR transcription. Interestingly, the dominant negative form of Rac1 (RacN17) was also efficient in relieving FasR expression, suggesting that the increase in FasR expression following AKT stimuli could be mediated via AKT ability to elicit suppression of Rac1, which in turn decreases JNK activities and c-Jun phosphorylation. Overall, our findings demonstrate that through its negative effects on both c-Jun and STAT3, the PI3K-AKT pathway disrupts cooperation between c-Jun and STAT3, which is required for silencing the FasR promoter, resulting in increased expression of surface FasR and concomitant sensitization to FasL-mediated programmed cell death. PMID- 11733517 TI - Inactivation of caspase-8 on mitochondria of Bcl-xL-expressing MCF7-Fas cells: role for the bifunctional apoptosis regulator protein. AB - Apoptosis induction through CD95 (APO-1/Fas) critically depends on generation of active caspase-8 at the death-inducing signaling complex (DISC). Depending on the cell type, active caspase-8 either directly activates caspase-3 (type I cells) or relies on mitochondrial signal amplification (type II cells). In MCF7-Fas cells that are deficient for pro-caspase-3, even high amounts of caspase-8 produced at the DISC cannot directly activate downstream effector caspases without mitochondrial help. Overexpression of Bcl-x(L) in these cells renders them resistant to CD95-mediated apoptosis. However, activation of caspase-8 in control (vector) and Bcl-x(L) transfectants of MCF7-Fas cells proceeds with similar kinetics, resulting in a complete processing of cellular caspase-8. Most of the cytosolic caspase-8 substrates are not cleaved in the Bcl-x(L) protected cells, raising the question of how Bcl-x(L)-expressing MCF7-Fas cells survive large amounts of potentially cytotoxic caspase-8. We now demonstrate that active caspase-8 is initially generated at the DISC of both MCF7-Fas-Vec and MCF7-Fas Bcl-x(L) cells and that the early steps of CD95 signaling such as caspase-8 dependent cleavage of DISC bound c-FLIP(L), caspase-8-dependent clustering, and internalization of CD95, as well as processing of pro-caspase-8 bound to mitochondria are very similar in both transfectants. However, events downstream of mitochondria, such as release of cytochrome c, only occur in the vector transfected MCF7-Fas cells, and no in vivo caspase-8 activity can be detected in the Bcl-x(L)-expressing cells. Our data suggest that, in Bcl-x(L)-expressing MCF7 Fas cells, active caspase-8 is sequestered on the outer mitochondrial surface presumably by association with the protein "bifunctional apoptosis regulator" in a way that does not allow substrates to be cleaved, identifying a novel mechanism of regulation of apoptosis sensitivity by mitochondrial Bcl-x(L). PMID- 11733518 TI - Use of indirect site-directed mutagenesis to alter the substrate specificity of methylamine dehydrogenase. AB - Methylamine dehydrogenase (MADH) is a tryptophan tryptophylquinone-dependent enzyme that catalyzes the oxidative deamination of primary amines. Native MADH exhibits a strong preference for methylamine over longer carbon chain amines. Residue alphaPhe(55) controls this substrate specificity. When alphaPhe(55) is replaced with Ala, this preference is reversed with alphaF55A MADH preferring long-chain amines with at least seven carbons (Zhu, Z., Sun, D., and Davidson, V. L. (2000) Biochemistry 39, 11184-11186). To further modulate the substrate specificity of MADH, the side-chain of alphaPhe(55) was repositioned by site directed mutagenesis of residue betaIle(107). This residue makes close contact with alphaPhe(55) and restricts its movement. betaI107V MADH exhibits a strong preference for propylamine, and betaI107N MADH exhibits a preference for 1 aminopentane. Thus, it has been possible to create forms of MADH that exhibit a preference for amines with carbon chain lengths of one, three, five, or seven carbons. The ability to discriminate between amines of different chain length was essentially abolished by an alphaF55I mutation. Molecular modeling studies with the known crystal structure are described that provide an explanation for these results. These results provide an example of a design-based approach to protein engineering in which site-directed mutagenesis on one residue can be used to reposition another residue to specifically alter enzyme specificity. PMID- 11733519 TI - Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-protein complex. AB - We previously demonstrated that in Chinese hamster ovary cells scavenger receptor, class B, type I-dependent selective cholesteryl ester uptake occurs in caveolae. In the present study we hypothesized that cholesteryl ester is transported from caveolae through the cytosol to an internal membrane by a caveolin chaperone complex similar to the one we originally described for the transport of newly synthesized cholesterol. To test this hypothesis we incubated Chinese hamster ovary cells expressing scavenger receptor, class B, type I with [(3)H]cholesteryl ester-labeled high density lipoprotein, subfractionated the cells and looked for a cytosolic pool of [(3)H]cholesteryl ester. The radiolabeled sterol initially appeared in the caveolae fraction, then in the cytosol, and finally in the internal membrane fraction. Caveolin IgG precipitated all of the [(3)H]cholesteryl ester associated with the cytosol. Co immunoprecipitation studies demonstrated that in the presence of high density lipoprotein, but not low density lipoprotein or lipoprotein-deficient serum, caveolin IgG precipitated four proteins: annexin II, cyclophilin 40, caveolin, and cyclophilin A. Caveolin acylation-deficient mutants were used to demonstrate that acylation of cysteine 133 but not cysteine 143 or 156 is required for annexin II association with caveolin and the rapid transport of cholesteryl esters out of caveolae. We conclude that a caveolin-annexin II lipid-protein complex facilitates the rapid internalization of cholesteryl esters from caveolae. PMID- 11733520 TI - Activation of Paneth cell alpha-defensins in mouse small intestine. AB - Paneth cells in small intestine crypts secrete microbicidal alpha-defensins, termed cryptdins, as components of enteric innate immunity. The bactericidal activity of cryptdins requires proteolytic activation of precursors by matrix metalloproteinase-7 (MMP-7; matrilysin) (Wilson, C. L., Ouellette, A. J., Satchell, D. P., Ayabe, T., Lopez-Boado, Y. S., Stratman, J. L., Hultgren, S. J., Matrisian, L. M., and Parks, W. C. (1999) Science 286, 113-117). Here, we report on the intracellular processing of cryptdin proforms in mouse Paneth cells. Peptide sequencing of MMP-7 digests of purified natural procryptdins identified conserved cleavage sites in the proregion between Ser(43) and Val(44) as well as at the cryptdin peptide N terminus between Ser(58) and Leu(59). Immunostaining co localized precursor prosegments and mature cryptdin peptides to Paneth cell granules, providing evidence of their secretion. Extensive MMP-7-dependent procryptdin processing occurs in Paneth cells, as shown by Western blot analyses of intestinal crypt proteins and proteins from granule-enriched subcellular fractions. The addition of soluble prosegments to in vitro antimicrobial peptide assays inhibited the bactericidal activities of cryptdin-3 and -4 in trans, suggesting possible cytoprotective effects by prosegments prior to secretion. Levels of activated cryptdins were normal in small bowel of germ-free mice and in sterile implants of fetal mouse small intestine grown subcutaneously. Thus, the initiation of procryptdin processing by MMP-7 does not require direct bacterial exposure, and the basal MMP-7 content of germ-free Paneth cells is sufficient to process and activate alpha-defensin precursors. MMP-7-dependent procryptdin activation in vivo provides mouse Paneth cells with functional peptides for apical secretion into the small intestine lumen. PMID- 11733521 TI - Diversity of neuron-specific K+-Cl- cotransporter expression and inhibitory postsynaptic potential depression in rat motoneurons. AB - Motoneurons receive a robust recurrent synaptic inhibition by gamma-aminobutyric acid and glycine, which activate Cl(-) channels. Thus, Cl(-) homeostasis determines the efficacy of synaptic inhibition in the motoneurons. In situ hybridization reveals that the neuronal K(+)-Cl(-) cotransporter isoform 2 (KCC2), a major mechanism in maintaining a low Cl(-) concentration in neurons, is abundantly expressed in the facial, hypoglossal (XII), and spinal motoneurons innervating striated muscle, whereas the dorsal vagal motoneurons (DMVs) controlling smooth muscle exhibited little expression of KCC2. This raises a general interest in the correlation between KCC2 expression and inhibitory postsynaptic potential (IPSP) performance in the native circuits. Intracellular and whole-cell patch recordings revealed that an activity-dependent depression of IPSPs and positive shift of IPSP reversal potentials were more prominent in the DMV than in the XII. Cl(-) influx through Cl(-) channels was extruded more potently in the XII than in the DMV, suggesting that differences in Cl(-) extrusion account for these dynamic differences of IPSP. Cl(-) extrusion was inhibited by either furosemide or an increase in extracellular potassium concentrations. Thus, the rigid maintenance of IPSP and rapid Cl(-) extrusion in the XII reflects an intense expression of KCC2. KCC2 expression may strongly influence the IPSP depression and functional properties of the motoneurons innervating striated muscles. PMID- 11733522 TI - Mapping of functional domains in the p22(phox) subunit of flavocytochrome b(559) participating in the assembly of the NADPH oxidase complex by "peptide walking". AB - The superoxide-generating NADPH oxidase complex of phagocytes consists of a membranal heterodimeric flavocytochrome (cytochrome b(559)), composed of gp91(phox) and p22(phox) subunits, and four cytosolic proteins, p47(phox), p67(phox), p40(phox), and the small GTPase Rac (1 or 2). All redox stations involved in electron transport from NADPH to oxygen are located in gp91(phox). NADPH oxidase activation is the consequence of assembly of cytochrome b(559) with cytosolic proteins, a process reproducible in a cell-free system, consisting of phagocyte membranes, and recombinant cytosolic components, activated by an anionic amphiphile. p22(phox) is believed to act as a linker between the cytosolic components and gp91(phox). We applied "peptide walking" to mapping of domains in p22(phox) participating in NADPH oxidase assembly. Ninety one synthetic overlapping pentadecapeptides, spanning the p22(phox) sequence, were tested for the ability to inhibit NADPH oxidase activation in the cell-free system and to bind individual cytosolic NADPH oxidase components. We conclude the following. 1) The p22(phox) subunit of cytochrome b(559) serves as an anchor for both p47(phox) and p67(phox). 2) p47(phox) binds not only to the proline-rich region, located at residues 151-160 in the cytosolic C terminus of p22(phox), but also to a domain (residues 51-63) located on a loop exposed to the cytosol. 3) p67(phox) shares with p47(phox) the ability to bind to the proline-rich region (residues 151-160) and also binds to two additional domains, in the cytosolic loop (residues 81-91) and at the start of the cytosolic tail (residues 111-115). 4) The binding affinity of p67(phox) for p22(phox) peptides is lower than that of p47(phox). 5) Binding of both p47(phox) and p67(phox) to proline-rich p22(phox) peptides occurs in the absence of an anionic amphiphile. A revised membrane topology model of p22(phox) is proposed, the core of which is the presence of a functionally important cytosolic loop (residues 51-91). PMID- 11733523 TI - Axonin-1/TAG-1 mediates cell-cell adhesion by a cis-assisted trans-interaction. AB - The neural cell adhesion molecule axonin-1/TAG-1 mediates cell-cell interactions via homophilic and heterophilic contacts. It consists of six Ig and four fibronectin type III domains anchored to the membrane by glycosylphosphatidylinositol. The recently solved crystal structure indicates a module composed of the four N-terminal Ig domains as the contact site between trans-interacting axonin-1 molecules from apposed membranes. Here, we have tested domain-specific monoclonal antibodies for their capacity to interfere with homophilic binding in a cell aggregation assay. The results confirmed the existence of a binding region within the N-terminal Ig domains and identified a second region contributing to homophilic binding on the third and fourth fibronectin domains near the C terminus. The perturbation of each region alone resulted in a complete loss of cell aggregation, suggesting that axonin-1 mediated cell-cell contact results from a cooperative action of two homophilic binding regions. The data support that axonin-1-mediated cell-cell contact is formed by cis-assisted trans-binding. The N-terminal binding regions of axonin-1 establish a linear zipper-like string of trans-interacting axonin-1 molecules alternately provided by the two apposed membranes. The C-terminal binding regions strengthen the cell-cell contact by enhancing the expansion of the linear string into a two-dimensional array via cis-interactions. Cis-assisted trans-binding may be a basic binding mechanism common to many cell adhesion molecules. PMID- 11733524 TI - Methylation-mediated silencing of TMS1/ASC is accompanied by histone hypoacetylation and CpG island-localized changes in chromatin architecture. AB - Aberrant methylation of CpG-dense islands in the promoter regions of genes is an acquired epigenetic alteration associated with the silencing of tumor suppressor genes in human cancers. In a screen for endogenous targets of methylation mediated gene silencing, we identified a novel CpG island-associated gene, TMS1, which is aberrantly methylated and silenced in response to the ectopic expression of DNA methyltransferase-1. TMS1 functions in the regulation of apoptosis and is frequently methylated and silenced in human breast cancers. In this study, we characterized the methylation pattern and chromatin architecture of the TMS1 locus in normal fibroblasts and determined the changes associated with its progressive methylation. In normal fibroblasts expressing TMS1, the CpG island is defined by an unmethylated domain that is separated from densely methylated flanking DNA by distinct 5' and 3' boundaries. Analysis of the nucleoprotein architecture of the locus in intact nuclei revealed three DNase I-hypersensitive sites that map within the CpG island. Strikingly, two of these sites coincided with the 5'- and 3'-methylation boundaries. Methylation of the TMS1 CpG island was accompanied by loss of hypersensitive site formation, hypoacetylation of histones H3 and H4, and gene silencing. This altered chromatin structure was confined to the CpG island and occurred without significant changes in methylation, histone acetylation, or hypersensitive site formation at a fourth DNase I-hypersensitive site 2 kb downstream of the TMS1 CpG island. The data indicate that there are sites of protein binding and/or structural transitions that define the boundaries of the unmethylated CpG island in normal cells and that aberrant methylation overcomes these boundaries to direct a local change in chromatin structure, resulting in gene silencing. PMID- 11733525 TI - Reversible and specific extracellular antagonism of receptor-histidine kinase signaling. AB - Staphylococcal pathogenesis is regulated by a two-component quorum-sensing system, agr, activated by a self-coded autoinducing peptide (AIP). The agr system is widely divergent and is unique in that variant AIPs cross-inhibit agr activation in heterologous combinations. Cross-inhibition, but not self activation, is widely tolerant of structural diversity in the AIPs so that these two processes must involve different mechanisms of interaction with the respective receptors. Herein, we have utilized this naturally occurring antagonism to demonstrate that both activation and inhibition are reversible and that activators and inhibitors interact at a common site on the receptor. These results suggest that molecules designed to compete with natural agonists for binding at receptor-histidine kinase sensor domains could represent a general approach to the inhibition of receptor-histidine kinase signaling. PMID- 11733526 TI - Resolving a fidelity paradox: why Escherichia coli DNA polymerase II makes more base substitution errors in AT- compared with GC-rich DNA. AB - The activity of DNA polymerase-associated proofreading 3'-exonucleases is generally enhanced in less stable DNA regions leading to a reduction in base substitution error frequencies in AT- versus GC-rich sequences. Unexpectedly, however, the opposite result was found for Escherichia coli DNA polymerase II (pol II). Nucleotide misincorporation frequencies for pol II were found to be 3-5 fold higher in AT- compared with GC-rich DNA, both in the presence and absence of polymerase processivity subunits, beta dimer and gamma complex. In contrast, E. coli pol III holoenzyme, behaving "as expected," exhibited 3-5-fold lower misincorporation frequencies in AT-rich DNA. A reduction in fidelity in AT-rich regions occurred for pol II despite having an associated 3'-exonuclease proofreading activity that preferentially degrades AT-rich compared with GC-rich DNA primer-template in the absence of DNA synthesis. Concomitant with a reduction in fidelity, pol II polymerization efficiencies were 2-6-fold higher in AT-rich DNA, depending on sequence context. Pol II paradoxical fidelity behavior can be accounted for by the enzyme's preference for forward polymerization in AT-rich sequences. The more efficient polymerization suppresses proofreading thereby causing a significant increase in base substitution error rates in AT-rich regions. PMID- 11733527 TI - Changing the substrate reactivity of 2-hydroxybiphenyl 3-monooxygenase from Pseudomonas azelaica HBP1 by directed evolution. AB - The substrate reactivity of the flavoenzyme 2-hydroxybiphenyl 3-monooxygenase (EC, HbpA) was changed by directed evolution using error-prone PCR. In situ screening of mutant libraries resulted in the identification of proteins with increased activity towards 2-tert-butylphenol and guaiacol (2-methoxyphenol). One enzyme variant contained amino acid substitutions V368A/L417F, which were inserted by two rounds of mutagenesis. The double replacement improved the efficiency of substrate hydroxylation by reducing the uncoupled oxidation of NADH. With guaiacol as substrate, the two substitutions increased V(max) from 0.22 to 0.43 units mg(-1) protein and decreased the K'(m) from 588 to 143 microm, improving k'(cat)/K'(m) by a factor of 8.2. With 2-tert-butylphenol as the substrate, k'(cat) was increased more than 5-fold. Another selected enzyme variant contained amino acid substitution I244V and had a 30% higher specific activity with 2-sec-butylphenol, guaiacol, and the "natural" substrate 2 hydroxybiphenyl. The K'(m) for guaiacol decreased with this mutant, but the K'(m) for 2-hydroxybiphenyl increased. The primary structure of HbpA shares 20.1% sequence identity with phenol 2-monooxygenase from Trichosporon cutaneum. Structure homology modeling with this three-domain enzyme suggests that Ile(244) of HbpA is located in the substrate binding pocket and is involved in accommodating the phenyl substituent of the phenol. In contrast, Val(368) and Leu(417) are not close to the active site and would not have been obvious candidates for modification by rational design. PMID- 11733528 TI - The conserved Mynd domain of BS69 binds cellular and oncoviral proteins through a common PXLXP motif. AB - BS69 is a transcriptional co-repressor protein and a potential tumor suppressor that binds to the adenoviral oncoprotein E1A. We show that the C-terminal Mynd domain of BS69 (amino acids 516-561) or the closely related Mynd domains of the Caenorhabditis elegans proteins Bra-1 and Bra-2 bind not only to E1A but also to the Epstein-Barr virus EBNA2 oncoprotein and the Myc-related cellular protein MGA. Interaction depends on intact PXLXP motifs present in all three proteins. Moreover, viral proteins compete for binding of BS69 to MGA in a PXLXP-dependent fashion. Because deletions in E1A or EBNA2 that cover the PXLXP motifs are non transforming, our observations suggest a role for BS69 in cell growth control that is reminiscent of abrogation of the Rb function by various oncoproteins. PMID- 11733529 TI - Signal transduction through the B cell antigen receptor is normal in ataxia telangiectasia B lymphocytes. AB - The rare human genetic disorder ataxia-telangiectasia (A-T) has multiple consequences including a variable degree of immunodeficiency. Khanna and co workers (Khanna, K. K., Yan, J., Watters, D., Hobson, K., Beamish, H., Spring, K., Shiloh, Y., Gatti, R. A., and Lavin, M. F. (1997) J. Biol. Chem. 272, 9489 9495) evaluated signaling in Epstein-Barr virus (EBV) immortalized A-T lymphoblastoid cell lines (LCLs), derived from the B cells of A-T patients. They showed that A-T lymphoblastoid cells lack signaling through the B cell antigen receptor and concluded that the fault in A-T encompasses intracellular signaling in B cells. However, it is established that EBV latent membrane protein 2A (LMP2A) blocks signaling in EBV-bearing cells by interaction with cellular tyrosine kinases. To test whether the reported fault in A-T B cells was not inherent in A-T but the result of influence of wild-type EBV, we derived A-T LCLs with wild-type or LMP2A-deleted EBV and studied signaling in these cells in response to cross-linking the B cell antigen receptor. We report that intracellular calcium mobilization and tyrosine phosphorylation in LMP2A-depleted LCLs derived from A-T patients is indistinguishable from that in LMP2A-depleted LCLs derived from normal controls. Further, signaling is blocked similarly in A-T and normal lymphoblastoid cells bearing wild-type EBV. In conclusion there is no evidence of any defect in B cell receptor signal transduction in A-T B cells. PMID- 11733530 TI - The domain organization and properties of individual domains of DNA topoisomerase V, a type 1B topoisomerase with DNA repair activities. AB - Topoisomerase V (Topo V) is a type IB (eukaryotic-like) DNA topoisomerase. It was discovered in the hyperthermophilic prokaryote Methanopyrus kandleri and is the only topoisomerase with associated apurinic/apyrimidinic (AP) site-processing activities. The structure of Topo V in the free and DNA-bound states was probed by limited proteolysis at 37 degrees C and 80 degrees C. The Topo V protein is comprised of (i) a 44-kDa NH(2)-terminal core subdomain, which contains the active site tyrosine residue for topoisomerase activity, (ii) an immediately adjacent 16-kDa subdomain that contains degenerate helix-hairpin-helix (HhH) motifs, (iii) a protease-sensitive 18-kDa HhH "hinge" region, and (iv) a 34-kDa COOH-terminal HhH domain. Three truncated Topo V polypeptides comprising the NH(2)-terminal 44-kDa and 16-kDa domains (Topo61), the 44-, 16-, and 18-kDa domains (Topo78), and the COOH-terminal 34-kDa domain (Topo34) were cloned, purified, and characterized. Both Topo61 and Topo78 are active topoisomerases, but in contrast to Topo V these enzymes are inhibited by high salt concentrations. Topo34 has strong DNA-binding ability but shows no topoisomerase activity. Finally, we demonstrate that Topo78 and Topo34 possess AP lyase activities that are important in base excision DNA repair. Thus, Topo V has at least two active sites capable of processing AP DNA. The significance of multiple HhH motifs for the Topo V processivity is discussed. PMID- 11733531 TI - JEAP, a novel component of tight junctions in exocrine cells. AB - Tight junctions (TJs) consist of transmembrane proteins and many peripheral membrane proteins. To further characterize the molecular organization of TJs, we attempted here to screen for novel TJ proteins by the fluorescence localization based expression cloning method. We identified a novel peripheral membrane protein at TJs and named it junction-enriched and -associated protein (JEAP). JEAP consists of 882 amino acids with a calculated molecular weight of 98,444. JEAP contained a polyglutamic acid repeat at the N-terminal region, a coiled-coil domain at the middle region, and a consensus motif for binding to PDZ domains at the C-terminal region. Exogenously expressed JEAP co-localized with ZO-1 and occludin at TJs in polarized Madin-Darby canine kidney cells, but not with claudin-1, JAM, or ZO-1 in L cells. Endogenous JEAP localized at TJs of exocrine cells including pancreas, submandibular gland, lacrimal gland, parotid gland, and sublingual gland, but not at TJs of epithelial cells of small intestine or endothelial cells of blood vessels. The present results indicate that JEAP is a novel component of TJs, which is specifically expressed in exocrine cells. PMID- 11733532 TI - The HET-s prion protein of the filamentous fungus Podospora anserina aggregates in vitro into amyloid-like fibrils. AB - The HET-s protein of Podospora anserina is a fungal prion. This protein behaves as an infectious cytoplasmic element that is transmitted horizontally from one strain to another. Under the prion form, the HET-s protein forms aggregates in vivo. The specificity of this prion model compared with the yeast prions resides in the fact that under the prion form HET-s causes a growth inhibition and cell death reaction when co-expressed with the HET-S protein from which it differs by 13 residues. Herein we describe the purification and initial characterization of recombinant HET-s protein expressed in Escherichia coli. The HET-s protein self associates over time into high molecular weight aggregates. These aggregates greatly accelerate precipitation of the soluble form. HET-s aggregates appear as amyloid-like fibrils using electron microscopy. They bind Congo Red and show birefringence under polarized light. In the aggregated form, a HET-s fragment of approximately 7 kDa is resistant to proteinase K digestion. CD and FTIR analyses indicate that upon transition to the aggregated state, the HET-s protein undergoes a structural rearrangement characterized by an increase in antiparallel beta-sheet structure content. These results suggest that the [Het-s] prion element propagates in vivo as an infectious amyloid. PMID- 11733533 TI - Strain-based genetic differences regulate the efficiency of systemic gene delivery as well as expression. AB - We have characterized the impact of strain-based genetic differences on the efficiency of the intravenous cationic liposome-DNA complex (CLDC)-based gene transfer and expression in mice. We also investigated what steps in the gene delivery and expression pathway appeared responsible for these strain-related differences and whether such differences could be compensated for either by agents that alter host pathways important in CLDC-mediated gene transfer and expression, or by changes in CLDC formulation. We found that different mouse strains can exhibit different expression levels and/or differences in the amount of plasmid DNA delivered to the organs where the DNA is expressed. Furthermore, drug pretreatment or reformulation of the CLDC could improve DNA delivery and/or gene expression in a strain-specific fashion. We conclude that genetic factors critically modify both the tissue deposition and the expression of genetic materials delivered by CLDC. Because manipulation of either the host or the CLDC could at least partially compensate for these strain-related differences, such strategies may be required to effectively use non-viral gene transfer approaches in genetically diverse populations. PMID- 11733534 TI - Expression of full-length polyglutamine-expanded Huntingtin disrupts growth factor receptor signaling in rat pheochromocytoma (PC12) cells. AB - We reported previously that normal Huntingtin is associated with epidermal growth factor receptor (EGF) signaling complex (Liu, Y. F., Deth, C. R., and Devys, D. (1997) J. Biol. Chem. 272, 8121-8124). To investigate the potential role of normal and polyglutamine-expanded Huntingtin in the regulation of growth factor receptor-mediated cellular signaling and biological function, we stably transfected full-length Huntingtin containing 16, 48, or 89 polyglutamine repeats into PC12 cells where cellular signaling mechanisms, mediated by nerve growth factor (NGF) or EGF receptors, are well characterized. Expression of polyglutamine-expanded Huntingtin, but not normal Huntingtin, leads to a dramatic morphological change. In clones carrying the mutated Huntingtin, both NGF and EGF receptor-mediated activation of mitogen-activated protein kinase, c-Jun N terminal kinase, and Akt are significantly attenuated, and NGF receptor-mediated neurite outgrowth is blocked. Co-immunoprecipitation studies show that the associations of NGF or EGF receptors with growth factor receptor-binding protein 2 (Grb2) and phosphoinositide 3-kinase are significantly inhibited. NGF-induced tyrosine phosphorylation of NGF receptors (TrkA) is also consistently suppressed. Our data demonstrate that polyglutamine-expanded Huntingtin disrupts cellular signaling mediated by both EGF and NGF receptors in PC12 cells. It is known that Huntington's disease patients exhibit an extremely low incidence of a variety of cancers and are deficient in glucose metabolism. Thus, our results may reflect an important molecular mechanism for the pathogenesis of the disease. PMID- 11733535 TI - Active site mutations in DNA topoisomerase I distinguish the cytotoxic activities of camptothecin and the indolocarbazole, rebeccamycin. AB - DNA topoisomerase I (Top1p) catalyzes topological changes in DNA and is the cellular target of the antitumor agent camptothecin (CPT). Non-CPT drugs that target Top1p, such as indolocarbazoles, are under clinical development. However, whether the cytotoxicity of indolocarbazoles derives from Top1p poisoning remains unclear. To further investigate indolocarbazole mechanism, rebeccamycin R-3 activity was examined in vitro and in yeast. Using a series of Top1p mutants, where substitution of residues around the active site tyrosine has well-defined effects on enzyme catalysis, we show that catalytically active, CPT-resistant enzymes remain sensitive to R-3. This indolocarbazole did not inhibit yeast Top1p activity, yet was effective in stabilizing Top1p-DNA complexes. Similar results were obtained with human Top1p, when Ser or His were substituted for Asn-722. The mutations altered enzyme function and sensitivity to CPT, yet R-3 poisoning of Top1p was unaffected. Moreover, top1delta, rad52delta yeast cells expressing human Top1p, but not catalytically inactive Top1Y723Fp, were sensitive to R-3. These data support hTop1p as the cellular target of R-3 and indicate that distinct drug-enzyme interactions at the active site are required for efficient poisoning by R-3 or CPT. Furthermore, resistance to one poison may potentiate cell sensitivity to structurally distinct compounds that also target Top1p. PMID- 11733536 TI - Promoter sequences targeting tissue-specific gene expression of hypothalamic and ovarian gonadotropin-releasing hormone in vivo. AB - Molecular mechanisms directing tissue-specific expression of gonadotropin releasing hormone (GnRH) are difficult to study due to the paucity and scattered distribution of GnRH neurons. To identify regions of the mouse GnRH (mGnRH) promoter that are critical for appropriate tissue-specific gene expression, we generated transgenic mice with fragments (-3446/+23 bp, -2078/+23 bp, and 1005/+28 bp) of mGnRH promoter fused to the luciferase reporter gene. The pattern of mGnRH promoter activity was assessed by measuring luciferase activity in tissue homogenates. All three 5'-fragments of mGnRH promoter targeted hypothalamic expression of the luciferase transgene, but with the exception of the ovary, luciferase expression was absent in non-neural tissues. High levels of ovarian luciferase activity were observed in mice generated with both -2078 and 1005 bp of promoter. Our study is the first to define a region of the GnRH gene promoter that directs expression to both neural and non-neural tissues in vivo. We demonstrate that DNA sequences contained within the proximal -1005 bp of the mGnRH promoter are sufficient to direct mGnRH gene expression to both the ovary and hypothalamus. Our results also suggest that DNA sequences distal to -2078 bp mediate the repression of ovarian GnRH. PMID- 11733537 TI - Distinct roles of the Ikappa B kinase alpha and beta subunits in liberating nuclear factor kappa B (NF-kappa B) from Ikappa B and in phosphorylating the p65 subunit of NF-kappa B. AB - Phosphatidylinositol 3'-kinase (PI3K) and the serine/threonine kinase AKT have critical roles in phosphorylating and transactivating the p65 subunit of nuclear factor kappaB (NF-kappaB) in response to the pro-inflammatory cytokines interleukin-1 (IL-1) and tumor necrosis factor (TNF). Mouse embryo fibroblasts (MEFs) lacking either the alpha or beta subunit of IkappaB kinase (IKK) were deficient in NF-kappaB-dependent transcription following treatment with IL-1 or TNF. However, in contrast to IKKbeta-null MEFs, IKKalpha-null MEFs were not substantially defective in the cytokine-stimulated degradation of Ikappabetaalpha or in the nuclear translocation of NF-kappaB. The IKK complexes from IKKalpha- or IKKbeta-null MEFs were both deficient in PI3K-mediated phosphorylation of the transactivation domain of the p65 subunit of NF-kappaB in response to IL-1 and TNF, and constitutively activated forms of PI3K or AKT did not potentiate cytokine-stimulated activation of NF-kappaB in either IKKalpha- or IKKbeta-null MEFs. Collectively, these data indicate that, in contrast to IKKbeta, which is required for both NF-kappaB liberation and p65 phosphorylation, IKKalpha is required solely for the cytokine-induced phosphorylation and activation of the p65 subunit of NF-kappaB that are mediated by the PI3K/AKT pathway. PMID- 11733538 TI - Ym1 is a neutrophil granule protein that crystallizes in p47phox-deficient mice. AB - Crystals were discovered within the aged lung and at sites of chronic inflammation in a mouse model of chronic granulomatous disease. Following re crystallization at neutral pH, the crystals were identified as the chitinase-like protein Ym1, expressed in organs of the lymphoreticular system, the lung, and distal stomach. Ym1 was shown to be a neutrophil granule protein and to have weak beta-N-acetylglucosaminidase activity, indicating that it might contribute to the digestion of glycosaminoglycans. Crystal formation is likely to be a function of excess neutrophil turnover at sites of inflammation in the chronic granulomatous disease mouse. Failure to remove subcutaneous Ym1 crystals injected into knockout mice indicates that a failure of digestion may also contribute to crystallization. PMID- 11733539 TI - High molecular weight neurofilament proteins are physiological substrates of adduction by the lipid peroxidation product hydroxynonenal. AB - Protein adducts of the lipid peroxidation product trans-4-hydroxy-2-nonenal (HNE) are features of oxidative damage in neuronal cell bodies in Alzheimer's disease but are also seen in axons of normal as well as diseased individuals. In this study, focusing on the axons of the mouse sciatic nerve, we found that HNE adducts characterize axons of mice from birth to senility. Immunoblots of axonal proteins showed that HNE adducts are only detected in neurofilament heavy subunit (NFH) and, to a lesser extent, neurofilament medium subunit (NFM), both lysine rich proteins, consistent with the adducts being limited to lysine residues. In vitro, HNE treatment of permeabilized sciatic nerve showed the same specificity, i.e. NFH and NFM are the only proteins that reacted with HNE, providing they are phosphorylated. Quantitative immunoblot analysis of two strains of mice ages 1-33 months showed that the levels of HNE adducts on NFH are consistent throughout life. Additionally, mice transgenic for human superoxide dismutase-1 with G85R mutation show no difference in HNE adduction to NFH compared with controls. Taken together, these studies indicate that HNE adduction to NFH is physiological, and its constancy from birth to senility as well as its dependence on phosphorylation argues that NFH and NFM modification may play a role in protecting the membrane rich axon from toxic aldehydes resulting from oxidative damage. PMID- 11733540 TI - Altered thymidine metabolism due to defects of thymidine phosphorylase. AB - Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive human disease due to mutations in the thymidine phosphorylase (TP) gene. TP enzyme catalyzes the reversible phosphorolysis of thymidine to thymine and 2-deoxy-D-ribose 1-phosphate. We present evidence that thymidine metabolism is altered in MNGIE. TP activities in buffy coats were reduced drastically in all 27 MNGIE patients compared with 19 controls. All MNGIE patients had much higher plasma levels of thymidine than normal individuals and asymptomatic TP mutation carriers. In two patients, the renal clearance of thymidine was approximately 20% that of creatinine, and because hemodialysis demonstrated that thymidine is ultrafiltratable, most of the filtered thymidine is likely to be reabsorbed by the kidney. In vitro, fibroblasts from controls catabolized thymidine in medium; by contrast, MNGIE fibroblasts released thymidine. In MNGIE, severe impairment of TP enzyme activity leads to increased plasma thymidine. In patients who are suspected of having MNGIE, determination of TP activity in buffy coats and thymidine levels in plasma are diagnostic. We hypothesize that excess thymidine alters mitochondrial nucleoside and nucleotide pools leading to impaired mitochondrial DNA replication, repair, or both. Therapies to reduce thymidine levels may be beneficial to MNGIE patients. PMID- 11733541 TI - Myotubularin and MTMR2, phosphatidylinositol 3-phosphatases mutated in myotubular myopathy and type 4B Charcot-Marie-Tooth disease. AB - Myotubularin is the archetype of a family of highly conserved protein-tyrosine phosphatase-like enzymes. The myotubularin gene, MTM1, is mutated in the genetic disorder, X-linked myotubular myopathy. We and others have previously shown that myotubularin utilizes the lipid second messenger, phosphatidylinositol 3 phosphate (PI(3)P), as a physiologic substrate. We demonstrate here that the myotubularin-related protein MTMR2, which is mutated in the neurodegenerative disorder, type 4B Charcot-Marie-Tooth disease, is also highly specific for PI(3)P as a substrate. Furthermore, the MTM-related phosphatases MTMR1, MTMR3, and MTMR6 also dephosphorylate PI(3)P, suggesting that activity toward this substrate is common to all myotubularin family enzymes. A direct comparison of the lipid phosphatase activities of recombinant myotubularin and MTMR2 demonstrates that their enzymatic properties are indistinguishable, indicating that the lack of functional redundancy between these proteins is likely to be due to factors other than the utilization of different physiologic substrates. To this end, we have analyzed myotubularin and MTMR2 transcripts during induced differentiation of cultured murine C2C12 myoblasts and find that their expression is divergently regulated. In addition, myotubularin and MTMR2 enhanced green fluorescent protein fusion proteins exhibit overlapping but distinct patterns of subcellular localization. Finally, we provide evidence that myotubularin, but not MTMR2, can modulate the levels of endosomal PI(3)P. From these data, we conclude that the developmental expression and subcellular localization of myotubularin and MTMR2 are differentially regulated, resulting in their utilization of specific cellular pools of PI(3)P. PMID- 11733542 TI - Control of cholesterol turnover in the mouse. PMID- 11733543 TI - A novel p21-activated kinase binds the actin and microtubule networks and induces microtubule stabilization. AB - Coordination of the different cytoskeleton networks in the cell is of central importance for morphogenesis, organelle transport, and motility. The Rho family proteins are well characterized for their effects on the actin cytoskeleton, but increasing evidence indicates that they may also control microtubule (MT) dynamics. Here, we demonstrate that a novel Cdc42/Rac effector, X-p21-activated kinase (PAK)5, colocalizes and binds to both the actin and MT networks and that its subcellular localization is regulated during cell cycle progression. In transfected cells, X-PAK5 promotes the formation of stabilized MTs that are associated in bundles and interferes with MTs dynamics, slowing both the elongation and shrinkage rates and inducing long paused periods. X-PAK5 subcellular localization is regulated tightly, since coexpression with active Rac or Cdc42 induces its shuttling to actin-rich structures. Thus, X-PAK5 is a novel MT-associated protein that may communicate between the actin and MT networks during cellular responses to environmental conditions. PMID- 11733544 TI - Vesicular and nonvesicular transport of ceramide from ER to the Golgi apparatus in yeast. AB - Transport and sorting of lipids must occur with specific mechanisms because the membranes of intracellular organelles differ in lipid composition even though most lipid biosynthesis begins in the ER. In yeast, ceramide is synthesized in the ER and transferred to the Golgi apparatus where inositolphosphorylceramide (IPC) is formed. These two facts imply that ceramide can be transported to the Golgi independent of vesicular traffic because IPC synthesis still continues when vesicular transport is blocked in sec mutants. Nonvesicular IPC synthesis in intact cells is not affected by ATP depletion. Using an in vitro assay that reconstitutes the nonvesicular pathway for transport of ceramide, we found that transport is temperature and cytosol dependent but energy independent. Preincubation of ER and Golgi fractions together at 4 degrees C, where ceramide transport does not occur, rendered the transport reaction membrane concentration independent, providing biochemical evidence that ER-Golgi membrane contacts stimulate ceramide transport. A cytosolic protease-sensitive factor is required after establishment of ER-Golgi contacts. PMID- 11733545 TI - A role for Vps1p, actin, and the Myo2p motor in peroxisome abundance and inheritance in Saccharomyces cerevisiae. AB - In vivo time-lapse microscopy reveals that the number of peroxisomes in Saccharomyces cerevisiae cells is fairly constant and that a subset of the organelles are targeted and segregated to the bud in a highly ordered, vectorial process. The dynamin-like protein Vps1p controls the number of peroxisomes, since in a vps1Delta mutant only one or two giant peroxisomes remain. Analogous to the function of other dynamin-related proteins, Vps1p may be involved in a membrane fission event that is required for the regulation of peroxisome abundance. We found that efficient segregation of peroxisomes from mother to bud is dependent on the actin cytoskeleton, and active movement of peroxisomes along actin filaments is driven by the class V myosin motor protein, Myo2p: (a) peroxisomal dynamics always paralleled the polarity of the actin cytoskeleton, (b) double labeling of peroxisomes and actin cables revealed a close association between both, (c) depolymerization of the actin cytoskeleton abolished all peroxisomal movements, and (d) in cells containing thermosensitive alleles of MYO2, all peroxisome movement immediately stopped at the nonpermissive temperature. In addition, time-lapse videos showing peroxisome movement in wild-type and vps1Delta cells suggest the existence of various levels of control involved in the partitioning of peroxisomes. PMID- 11733546 TI - Microtubule-associated protein 1B: a neuronal binding partner for myelin associated glycoprotein. AB - Myelin-associated glycoprotein (MAG) is expressed in periaxonal membranes of myelinating glia where it is believed to function in glia-axon interactions by binding to a component of the axolemma. Experiments involving Western blot overlay and coimmunoprecipitation demonstrated that MAG binds to a phosphorylated neuronal isoform of microtubule-associated protein 1B (MAP1B) expressed in dorsal root ganglion neurons (DRGNs) and axolemma-enriched fractions from myelinated axons of brain, but not to the isoform of MAP1B expressed by glial cells. The expression of some MAP1B as a neuronal plasma membrane glycoprotein (Tanner, S.L., R. Franzen, H. Jaffe, and R.H. Quarles. 2000. J. Neurochem. 75:553-562.), further documented here by its immunostaining without cell permeabilization, is consistent with it being a binding partner for MAG on the axonal surface. Binding sites for a MAG-Fc chimera on DRGNs colocalized with MAP1B on neuronal varicosities, and MAG and MAP1B also colocalized in the periaxonal region of myelinated axons. In addition, expression of the phosphorylated isoform of MAP1B was increased significantly when DRGNs were cocultured with MAG-transfected COS cells. The interaction of MAG with MAP1B is relevant to the known role of MAG in affecting the cytoskeletal structure and stability of myelinated axons. PMID- 11733547 TI - Caveolin-deficient mice: insights into caveolar function human disease. PMID- 11733548 TI - Series introduction. The immuno-neuroendocrine interface. PMID- 11733549 TI - The maternal-neonatal neuro-immune interface: are there long-term implications for inflammatory or stress-related disease? PMID- 11733550 TI - A kinase in the life of the beta cell. PMID- 11733551 TI - It infects me, it infects me not: phenotypic switching in the fungal pathogen Cryptococcus neoformans. PMID- 11733552 TI - The clinical relevance of glycobiology. PMID- 11733553 TI - Caught flat-footed: podocyte damage and the molecular bases of focal glomerulosclerosis. PMID- 11733554 TI - Organ-specific autoimmunity in mice whose T cell repertoire is shaped by a single antigenic peptide. AB - Organ-specific autoimmune diseases have been postulated to be the result of T cell response against organ-specific self-peptides bound to MHC molecules. Contrary to this paradigm, we report here that transgenic mice lacking MHC class I expression and expressing an MHC class II I-A(b) molecule that presents only a single peptide (E alpha 52-68) spontaneously develops peripheral nervous system specific autoimmune disease with many of the histopathological features found in experimental allergic neuritis. Reciprocal bone marrow chimeras produced using susceptible and resistant lines revealed that bone marrow-derived cells determined disease susceptibility. While the expression of the I-A(b)-E alpha 52 68 complex in the periphery was readily detectable in both lines, its expression on thymic dendritic cells responsible for tolerance induction was markedly lower in the susceptible line than in the resistant line. Consistent with this, CD4(+) T cells that can be activated by the I-A(b)-E alpha 52-68 complex were found in the susceptible line, but not in the resistant line. Such CD4(+) T cells conferred the disease to the resistant line by adoptive transfer, and administration of Ab specific for the I-A(b)-E alpha 52-68 complex inhibited disease manifestation in the susceptible line. These results indicate that disease development involves systemic T cell reactivity to I-A(b)-E alpha 52-68 complex, probably caused by incomplete negative thymocyte selection. PMID- 11733555 TI - Supramaximal cholecystokinin displaces Munc18c from the pancreatic acinar basal surface, redirecting apical exocytosis to the basal membrane. AB - Exocytosis at the apical surface of pancreatic acinar cells occurs in the presence of physiological concentrations of cholecystokinin (CCK) but is inhibited at high concentrations. Here we show that Munc18c is localized predominantly to the basal membranes of acinar cells. Supramaximal but not submaximal CCK stimulation caused Munc18c to dissociate from the plasma membrane, and this displacement was blocked by protein kinase C (PKC) inhibitors. Conversely, whereas the CCK analog CCK-OPE alone failed to displace Munc18c from the membrane, this agent caused Munc18c displacement following minimal PKC activation. To determine the physiological significance of this displacement, we used the fluorescent dye FM1-43 to visualize individual exocytosis events in real time from rat acinar cells in culture. We showed that supramaximal CCK inhibition of secretion resulted from impaired apical secretion and a redirection of exocytic events to restricted basal membrane sites. In contrast, CCK-OPE evoked apical exocytosis and could only induce basolateral exocytosis following activation of PKC. Infusion of supraphysiological concentrations of CCK in rats, a treatment that induced tissue changes reminiscent of mild acute pancreatitis, likewise resulted in rapid displacement of Munc18c from the basal membrane in vivo. PMID- 11733556 TI - A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If). AB - We describe a new congenital disorder of glycosylation, CDG-If. The patient has severe psychomotor retardation, seizures, failure to thrive, dry skin and scaling with erythroderma, and impaired vision. CDG-If is caused by a defect in the gene MPDU1, the human homologue of hamster Lec35, and is the first disorder to affect the use, rather than the biosynthesis, of donor substrates for lipid-linked oligosaccharides. This leads to the synthesis of incomplete and poorly transferred precursor oligosaccharides lacking both mannose and glucose residues. The patient has a homozygous point mutation (221T-->C, L74S) in a semiconserved amino acid of MPDU1. Chinese hamster ovary Lec35 cells lack a functional Lec35 gene and synthesize truncated lipid-linked oligosaccharides similar to the patient's. They lack glucose and mannose residues donated by Glc-P-Dol and Man-P Dol. Transfection with the normal human MPDU1 allele nearly completely restores normal glycosylation, whereas transfection with the patient's MPDU1 allele only weakly restores normal glycosylation. This work provides a new clinical picture for another CDG that may involve synthesis of multiple types of glycoconjugates. PMID- 11733557 TI - Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. AB - NPHS2 was recently identified as a gene whose mutations cause autosomal recessive steroid-resistant nephrotic syndrome. Its product, podocin, is a new member of the stomatin family, which consists of hairpin-like integral membrane proteins with intracellular NH(2)- and COOH-termini. Podocin is expressed in glomerular podocytes, but its subcellular distribution and interaction with other proteins are unknown. Here we show, by immunoelectron microscopy, that podocin localizes to the podocyte foot process membrane, at the insertion site of the slit diaphragm. Podocin accumulates in an oligomeric form in lipid rafts of the slit diaphragm. Moreover, GST pull-down experiments reveal that podocin associates via its COOH-terminal domain with CD2AP, a cytoplasmic binding partner of nephrin, and with nephrin itself. That podocin interacts with CD2AP and nephrin in vivo is shown by coimmunoprecipitation of these proteins from glomerular extracts. Furthermore, in vitro studies reveal direct interaction of podocin and CD2AP. Hence, as with the erythrocyte lipid raft protein stomatin, podocin is present in high-order oligomers and may serve a scaffolding function. We postulate that podocin serves in the structural organization of the slit diaphragm and the regulation of its filtration function. PMID- 11733558 TI - Islet beta cell expression of constitutively active Akt1/PKB alpha induces striking hypertrophy, hyperplasia, and hyperinsulinemia. AB - The phosphoinositide 3-kinase-Akt/PKB pathway mediates the mitogenic effects various nutrients and growth factors in cultured cells. To study its effects in vivo in pancreatic islet beta cells, we created transgenic mice that expressed a constitutively active Akt1/PKB alpha linked to an Insulin gene promoter. Transgenic mice exhibited a grossly visible increase in islet mass, largely due to proliferation of insulin-containing beta cells. Morphometric analysis verified a six-fold increase in beta cell mass/pancreas, a two-fold increase in 5-bromo-2' deoxyuridine incorporation, a four-fold increase in the number of beta cells per pancreas area, and a two-fold increase in cell size in transgenic compared with wild-type mice at 5 weeks. At least part of the increase in beta cell number may be accounted for by neogenesis, defined by criteria that include beta cells proliferating from ductular epithelium, and by a six-fold increase in the number of single and doublet beta cells scattered throughout the exocrine pancreas of the transgenic mice. Glucose tolerance was improved, and fasting as well as fed insulin was greater compared with wild-type mice. Glucose-stimulated insulin secretion was maintained in transgenic mice, which were resistant to streptozotocin-induced diabetes. We conclude that activation of the Akt1/PKB alpha pathway affects islet beta cell mass by alteration of size and number. PMID- 11733559 TI - Phenotypic switching of Cryptococcus neoformans occurs in vivo and influences the outcome of infection. AB - Phenotypic switching has been linked to the virulence of many pathogens, including fungi. However, it has not been conclusively shown to occur in vivo or to influence the outcome of infection. Cryptococcus neoformans undergoes phenotypic switching in vitro to colony types that differ in their virulence in mice. In this study, we asked whether C. neoformans undergoes phenotypic switching in vivo and whether this phenomenon contributes to virulence. By using a small inoculum to preclude the introduction of variants that had already switched during in vitro propagation, we demonstrated that in vivo switching to a mucoid phenotype occurred in two mice strains and was associated with a lethal outcome. Phenotypic switching resulted in changes of the capsular polysaccharide that inhibited phagocytosis by alveolar macrophages. This promoted a more vigorous inflammatory response and rapid demise. These data document in vivo switching in a fungus and associate this phenomenon with enhanced virulence and a lethal outcome. The importance of this finding is underscored by the increased likelihood of phenotypic switching in chronic cryptococcosis; thus this mechanism may account for the inability to eradicate the organism in immunocompromised hosts. PMID- 11733560 TI - Glomerulosclerosis is transmitted by bone marrow-derived mesangial cell progenitors. AB - We found that ROP Os/+ (Os/+) mice had diffuse glomerulosclerosis and glomerular hypertrophy and that their mesangial cells (the vascular smooth muscle cells of the glomerulus) displayed an apparent sclerosing phenotype. Since mesangial cells are the major source of scar tissue in glomerulosclerosis, we postulated that the sclerosis phenotype was carried by mesangial cell progenitors and that this phenotype could be derived from the bone marrow (BM). Therefore, we transplanted BM from Os/+ mice into congenic ROP +/+ mice (+/+ mice), which have normal glomeruli. We found that glomeruli of +/+ recipients of Os/+ marrow contained the Os/+ genotype, were hypertrophied, and contained increased extracellular matrix. Clones of recipient glomerular mesangial cells with the donor genotype were found in all +/+ recipients that developed mesangial sclerosis and glomerular hypertrophy, whereas +/+ recipients of +/+ BM had normal glomeruli. Thus, the sclerotic (Os/+) or normal (+/+) genotype and phenotype were present in, and transmitted by, BM-derived progenitors. These data show that glomerular mesangial cell progenitors are derived from the BM and can deliver a disease phenotype to normal glomeruli. Glomerular lesions may therefore be perpetuated or aggravated, rather than resolved, by newly arriving progenitor cells exhibiting a disease phenotype. PMID- 11733561 TI - Altered expression of the mRNA stability factor HuR promotes cyclooxygenase-2 expression in colon cancer cells. AB - Cyclooxygenase-2 (COX-2) expression is normally tightly regulated. However, constitutive overexpression plays a key role in colon carcinogenesis. To understand the molecular nature of enhanced COX-2 expression detected in colon cancer, we examined the ability of the AU-rich element-containing (ARE containing) 3' untranslated region (3'UTR) of COX-2 mRNA to regulate rapid mRNA decay in human colon cancer cells. In tumor cells displaying enhanced growth and tumorigenicity that is correlated with elevated COX-2, vascular endothelial growth factor (VEGF), and IL-8 protein levels, the corresponding mRNAs were transcribed constitutively and turned over slowly. The observed mRNA stabilization is owing to defective recognition of class II-type AREs present within the COX-2, VEGF, and IL-8 3'UTRs; c-myc mRNA, containing a class I ARE decayed rapidly in the same cells. Correlating with cellular defects in mRNA stability, the RNA-binding of trans-acting cellular factors was altered. In particular, we found that the RNA-stability factor HuR binds to the COX-2 ARE, and overexpression of HuR, as detected in tumors, results in elevated expression of COX-2, VEGF, and IL-8. These findings demonstrate the functional significance rapid mRNA decay plays in controlling gene expression and show that dysregulation of these trans-acting factors can lead to overexpression of COX-2 and other angiogenic proteins, as detected in neoplasia. PMID- 11733562 TI - Peroxisome proliferator-activated receptor-gamma haploinsufficiency enhances B cell proliferative responses and exacerbates experimentally induced arthritis. AB - Peroxisome proliferator-activated receptor-gamma (PPAR gamma) controls adipogenesis and glucose metabolism. It was reported recently that PPAR gamma activation by its agonistic ligands modifies lymphocyte function. Since synthetic ligands are known to exert their effect via PPAR gamma-dependent and -independent pathways, we examined the physiological role of PPAR gamma in lymphocytes by using heterozygote mutant mice in which one allele of PPAR gamma is deleted (PPAR gamma(+/-)). In contrast to T cells, which did not exhibit a significant difference, B cells from PPAR gamma(+/-) showed an enhanced proliferative response to stimulation by either lipopolysaccharide or cross-linking of antigen receptors. Dysregulation of the NF-kappa B pathway in B cells from PPAR gamma(+/ ) was indicated by spontaneous NF-kappa B activation, as well as increased I kappa B alpha phosphorylation and gel-shift activity following LPS stimulation. Mice primed with either ovalbumin or methylated BSA also showed enhanced antigen specific immune response of both T and B cells, an immunological abnormality that exacerbated antigen-induced arthritis. These findings indicate that PPAR gamma plays a critical role in the control of B cell response and imply a role in diseases in which B cell hyperreactivity is involved, such as arthritis and autoimmunity. PMID- 11733563 TI - High-affinity T helper epitope induces complementary helper and APC polarization, increased CTL, and protection against viral infection. AB - Natural viral proteins do not always make optimal vaccines. We have found that sequence modification to increase epitope affinity for class II MHC molecules (epitope enhancement) can improve immunogenicity. Here we show first that a higher-affinity helper epitope-enhanced HIV vaccine not only induces more cytotoxic T lymphocytes (CTLs), but also skews helper cells toward Th1 cytokine production and protects against HIV-1 recombinant vaccinia viral challenge. Furthermore, we elucidate a novel mechanism in which the higher-affinity vaccine induces dramatically more effective helper cells with a higher level of CD40L per helper cell and more positive cells, which in turn more effectively conditions dendritic cells (DCs) for CTL activation in a second culture. The improved helper cells also induce much greater IL-12 production by DCs, accounting for the reciprocal T helper polarization to Th1, and increase costimulatory molecule expression. Thus, increasing affinity for class II MHC results in a complementary interaction in which T helper and antigen-presenting cells polarize each other, as well as increase CTL, and provide greater vaccine efficacy against viral infection. PMID- 11733564 TI - MPDU1 mutations underlie a novel human congenital disorder of glycosylation, designated type If. AB - Deficiencies in the pathway of N-glycan biosynthesis lead to severe multisystem diseases, known as congenital disorders of glycosylation (CDG). The clinical appearance of CDG is variable, and different types can be distinguished according to the gene that is altered. In this report, we describe the molecular basis of a novel type of the disease in three unrelated patients diagnosed with CDG-I. Serum transferrin was hypoglycosylated and patients' fibroblasts accumulated incomplete lipid-linked oligosaccharide precursors for N-linked protein glycosylation. Transfer of incomplete oligosaccharides to protein was detected. Sequence analysis of the Lec35/MPDU1 gene, known to be involved in the use of dolichylphosphomannose and dolichylphosphoglucose, revealed mutations in all three patients. Retroviral-based expression of the normal Lec35 cDNA in primary fibroblasts of patients restored normal lipid-linked oligosaccharide biosynthesis. We concluded that mutations in the Lec35/MPDU1 gene cause CDG. This novel type was termed CDG-If. PMID- 11733565 TI - Bone marrow stromal cells mediate androgenic suppression of B lymphocyte development. AB - Castration of normal male mice induces expansion of the bone marrow B cell population, an effect that can be reversed by androgen replacement. We employed in vitro cultures and two in vivo models to investigate whether androgens exert these effects directly on marrow lymphoid precursors or whether actions on marrow stromal elements are required. Immature B cells from normal mouse bone marrow were not responsive to the suppressive effect of androgens unless they were cocultured with marrow stromal cells or with supernatants from androgen-treated stromal cells, suggesting that the androgen effects are exerted through marrow stromal elements by production of a diffusible mediator. Further experiments revealed that bone marrow stromal cells produced TGF-beta in response to dihydrotestosterone (DHT), and neutralization of TGF-beta in the DHT-treated stromal cells reversed the suppressive effects. The stromal cell requirement for androgen-mediated effects was confirmed in vivo by experiments using chimeric animals created by bone marrow transplantation in which androgen receptor expression was restricted to either the stromal or lymphoid cells of the bone marrow. Androgens only affected B cell development in chimeric mice with androgen sensitive stromal cells. These experiments suggest that effects of androgens on developing B cells are mediated through androgen receptors in bone marrow stromal cells. TGF-beta is a candidate mediator for these hormonal effects. PMID- 11733566 TI - Chloride conductance and genetic background modulate the cystic fibrosis phenotype of Delta F508 homozygous twins and siblings. AB - To investigate the impact of chloride (Cl(-)) permeability, mediated by residual activity of the cystic fibrosis transmembrane conductance regulator (CFTR) or by other Cl(-) channels, on the manifestations of cystic fibrosis (CF), we determined Cl(-) transport properties of the respiratory and intestinal tracts in Delta F508 homozygous twins and siblings. In the majority of patients, cAMP and/or Ca(2+)-regulated Cl(-) conductance was detected in the airways and intestine. Our finding of cAMP-mediated Cl(-) conductance suggests that, in vivo, at least some Delta F508 CFTR can reach the plasma membrane and affect Cl(-) permeability. In respiratory tissue, the expression of basal CFTR-mediated Cl(-) conductance, demonstrated by 30% of Delta F508 homozygotes, was identified as a positive predictor of milder CF disease. In intestinal tissue, 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid-insensitive (DIDS-insensitive) Cl( ) secretion, which is indicative of functional CFTR channels, correlated with a milder phenotype, whereas DIDS-sensitive Cl(-) secretion was observed mainly in more severely affected patients. The more concordant Cl(-) secretory patterns within monozygous twins compared with dizygous pairs imply that genes other than CFTR significantly influence the manifestation of the basic defect. PMID- 11733567 TI - Abnormal lipoprotein metabolism and reversible female infertility in HDL receptor (SR-BI)-deficient mice. AB - Mammalian female fertility depends on complex interactions between the ovary and the extraovarian environment (e.g., the hypothalamic-hypophyseal ovarian axis). The role of plasma lipoproteins in fertility was examined using HDL-receptor SR BI knockout (KO) mice. SR-BI KO females have abnormal HDLs, ovulate dysfunctional oocytes, and are infertile. Fertility was restored when the structure and/or quantity of abnormal HDL was altered by inactivating the apoAI gene or administering the cholesterol-lowering drug probucol. This suggests that abnormal lipoprotein metabolism can cause murine infertility--implying a functional hepatic-ovarian axis--and may contribute to some forms of human female infertility. PMID- 11733568 TI - The effect of salicylates on insulin sensitivity. PMID- 11733570 TI - Tumor rejection by disturbing tumor stroma cell interactions. AB - The stroma of solid tumors is a complex network of different cell types. We analyzed stroma cell interactions in two tumor models during cyclophosphamide (Cy)-induced tumor rejection. In growing tumors, tumor infiltrating macrophages (TIMs) produced interleukin (IL)-10. Beginning 6 h after Cy-treatment T cells in the tumor were inactivated and TIMs switched to interferon (IFN)-gamma production. Both, IL-10 production before and IFN-gamma production after Cy treatment by TIMs required T cells. With the same kinetics as TIMs started to produce IFN-gamma the tumor vasculature was destroyed which required IFN-gamma receptor expression on host but not tumor cells. These events preceded hemorrhagic necrosis and residual tumor cell elimination by T cells. Together, T cells regulate the function of TIMs and tumor rejection can be induced by disturbing the stroma network. PMID- 11733571 TI - Essential role of the prosurvival bcl-2 homologue A1 in mast cell survival after allergic activation. AB - Mast cells reside in tissues, where upon activation through the high-affinity-IgE receptor (FcepsilonRI) they degranulate and orchestrate the allergic reaction. Mast cells survive this activation and can thus be reactivated. In this study we demonstrate that this process depends on the pro-survival gene A1. Activation of mast cells through FcepsilonRI resulted in degranulation, strong induction of A1 mRNA and protein, and cell survival. In contrast, A1-deficient mast cells released granule mediators similar to the wild-type control, but the cells did not survive an allergic activation. Furthermore, A1(-/-) mice that had been sensitized and provoked with allergen exhibited a lower number of mast cell compared with littermate controls. The induction of A1 was dependent on calcium, as EDTA prevented A1 expression. The calcium ionophore, ionomycin, induced A1 expression and mast cell survival, whereas compound 48/80, a well-known mast cell secretagogue, did not. This study uncovers the importance of A1 for mast cell survival in allergic reactions, and it proposes A1 as a potential target for the treatment of allergic diseases. PMID- 11733572 TI - A Plasmodium falciparum homologue of Plasmodium vivax reticulocyte binding protein (PvRBP1) defines a trypsin-resistant erythrocyte invasion pathway. AB - Invasion of erythrocytes by Plasmodium merozoites is an intricate process involving multiple receptor-ligand interactions. The glycophorins and an unknown trypsin sensitive factor are all erythrocyte receptors used during invasion by the major human pathogen Plasmodium falciparum. However, only one erythrocyte receptor, Glycophorin A, has a well-established cognate parasite ligand, the merozoite protein erythrocyte binding antigen-175 (EBA-175). The involvement of several other parasite proteins during invasion have been proposed, but no direct evidence links them with a specific invasion pathway. Here we report the identification and characterization of P. falciparum normocyte binding protein 1 (PfNBP1), an ortholog of Plasmodium vivax reticulocyte binding protein-1. PfNBP1 binds to a sialic acid dependent trypsin-resistant receptor on the erythrocyte surface that appears to be distinct from known invasion receptors. Antibodies against PfNBP1 can inhibit invasion of trypsinized erythrocytes and two P. falciparum strains that express truncated PfNBP1 are unable to invade trypsinized erythrocytes. One of these strain, 7G8, also does not invade Glycophorin B negative erythrocytes. PfNBP1 therefore defines a novel trypsin-resistant invasion pathway and adds a level of complexity to current models for P. falciparum erythrocyte invasion. PMID- 11733573 TI - Ligand-independent signaling functions for the B lymphocyte antigen receptor and their role in positive selection during B lymphopoiesis. AB - Signal transduction through the B cell antigen receptor (BCR) is determined by a balance of positive and negative regulators. This balance is shifted by aggregation that results from binding to extracellular ligand. Aggregation of the BCR is necessary for eliciting negative selection or activation by BCR-expressing B cells. However, ligand-independent signaling through intermediate and mature forms of the BCR has been postulated to regulate B cell development and peripheral homeostasis. To address the importance of ligand-independent BCR signaling functions and their regulation during B cell development, we have designed a model that allows us to isolate the basal signaling functions of immunoglobulin (Ig)alpha/Igbeta-containing BCR complexes from those that are dependent upon ligand-mediated aggregation. In vivo, we find that basal signaling is sufficient to facilitate pro-B --> pre-B cell transition and to generate immature/mature peripheral B cells. The ability to generate basal signals and to drive developmental progression were both dependent on plasma membrane association of Igalpha/Igbeta complexes and intact immunoregulatory tyrosine activation motifs (ITAM), thereby establishing a correlation between these processes. We believe that these studies are the first to directly demonstrate biologically relevant basal signaling through the BCR where the ability to interact with both conventional as well as nonconventional extracellular ligands is eliminated. PMID- 11733574 TI - Isotype-specific selection of high affinity memory B cells in nasal-associated lymphoid tissue. AB - Mucosal immunoglobulin (Ig)A dominance has been proposed to be associated with preferential class switch recombination (CSR) to the IgA heavy chain constant region, Calpha. Here, we report that B cell activation in nasal-associated lymphoid tissue (NALT) upon stimulation with the hapten (4-hydroxy-3 nitrophenyl)acetyl (NP) coupled to chicken gamma globulin caused an anti-NP memory response dominated by high affinity IgA antibodies. In the response, however, NP-specific IgG(+) B cells expanded and sustained their number as a major population in germinal centers (GCs), supporting the view that CSR to IgG heavy chain constant region, Cgamma, operated efficiently in NALT. Both IgG(+) and IgA(+) GC B cells accumulated somatic mutations, indicative of affinity maturation to a similar extent, suggesting that both types of cell were equally selected by antigen. Despite the selection in GCs, high affinity NP-specific B cells were barely detected in the IgG memory compartment, whereas such cells dominated the IgA memory compartment. Taken together with the analysis of the V(H) gene clonotype in GC and memory B cells, we propose that NALT is equipped with a unique machinery providing IgA-specific enrichment of high affinity cells into the memory compartment, facilitating immunity with high affinity and noninflammatory secretory antibodies. PMID- 11733575 TI - Complement 1 inhibitor is a regulator of the alternative complement pathway. AB - We studied complement 1 inhibitor (C1-INH) as an inhibitor of the alternative complement pathway. C1-INH prevented lysis, induced by the alternative complement pathway, of paroxysmal nocturnal hemoglobinuria (PNH) erythrocytes in human serum. It inhibited the binding of both factors B and C3 to PNH and rabbit erythrocytes and blocked the ability of factor B to restore alternative-pathway function in factor B-depleted serum. C1-INH did not bind to factors B or D but did bind to immobilized C3b and cobra venom factor (CVF), a C3b analogue. C1-INH prevented factor B from binding to CVF-coated beads and dissociated bound factor B from such beads. Factor B and C1-INH showed cross competition in binding to CVF coated beads. Factor D cleaved factor B into Bb and Ba in the presence of C3b. Cleavage was markedly inhibited when C3b was preincubated with C1-INH. C1-INH inhibited the formation of CVFBb and decreased the C3 cleavage. Removal of C1-INH from serum, in the presence of Mg-EGTA with an anti-C1-INH immunoabsorbant, markedly increased alternative-pathway lysis. C1-INH interacts with C3b to inhibit binding of factor B to C3b. At physiologic concentrations, it is a downregulator of the alternative pathway convertase. PMID- 11733576 TI - A mediator role for metallothionein in tumor necrosis factor-induced lethal shock. AB - Tumor necrosis factor (TNF) is a proinflammatory cytokine, which is centrally involved in several inflammatory disorders. Administration of TNF leads to a potentially lethal systemic inflammatory response syndrome (SIRS). We observed that (a) mice lacking functional genes for metallothionein 1 and 2 (MT-null) were protected compared with wild-type controls (P = 0.0078), and (b) mice overexpressing MT-1 (MT-TG) were more sensitized for the lethal effect of TNF than control mice (P = 0.0003), indicating a mediating role for MT in TNF induced SIRS. As MT is involved in the body zinc homeostasis, we tested whether zinc deprivation or -supplementation alters the response to TNF. Although zinc depletion strongly sensitized (P = 0.036), and pretreatment with zinc sulfate (ZnSO4) conferred protection against the deleterious effects of TNF (P < 0.0002), it was also found that the protection provided by zinc is independent of MT. Our observation that hsp70 is strongly induced in jejunum after ZnSO4 treatment, suggests a contribution of hsp70 in the protection against TNF. In addition, ZnSO4 cotreatment allowed complete regression of inoculated tumors with TNF and interferon gamma, leading to a significantly better survival (P = 0.0045). PMID- 11733577 TI - Gene expression profiling of B cell chronic lymphocytic leukemia reveals a homogeneous phenotype related to memory B cells. AB - B cell-derived chronic lymphocytic leukemia (B-CLL) represents a common malignancy whose cell derivation and pathogenesis are unknown. Recent studies have shown that >50% of CLLs display hypermutated immunoglobulin variable region (IgV) sequences and a more favorable prognosis, suggesting that they may represent a distinct subset of CLLs which have transited through germinal centers (GCs), the physiologic site of IgV hypermutation. To further investigate the phenotype of CLLs, their cellular derivation and their relationship to normal B cells, we have analyzed their gene expression profiles using oligonucleotide based DNA chip microarrays representative of approximately 12,000 genes. The results show that CLLs display a common and characteristic gene expression profile that is largely independent of their IgV genotype. Nevertheless, a restricted number of genes (<30) have been identified whose differential expression can distinguish IgV mutated versus unmutated cases and identify them in independent panels of cases. Comparison of CLL profiles with those of purified normal B cell subpopulations indicates that the common CLL profile is more related to memory B cells than to those derived from naive B cells, CD5(+) B cells, and GC centroblasts and centrocytes. Finally, this analysis has identified a subset of genes specifically expressed by CLL cells of potential pathogenetic and clinical relevance. PMID- 11733578 TI - Relation of gene expression phenotype to immunoglobulin mutation genotype in B cell chronic lymphocytic leukemia. AB - The most common human leukemia is B cell chronic lymphocytic leukemia (CLL), a malignancy of mature B cells with a characteristic clinical presentation but a variable clinical course. The rearranged immunoglobulin (Ig) genes of CLL cells may be either germ-line in sequence or somatically mutated. Lack of Ig mutations defined a distinctly worse prognostic group of CLL patients raising the possibility that CLL comprises two distinct diseases. Using genomic-scale gene expression profiling, we show that CLL is characterized by a common gene expression "signature," irrespective of Ig mutational status, suggesting that CLL cases share a common mechanism of transformation and/or cell of origin. Nonetheless, the expression of hundreds of other genes correlated with the Ig mutational status, including many genes that are modulated in expression during mitogenic B cell receptor signaling. These genes were used to build a CLL subtype predictor that may help in the clinical classification of patients with this disease. PMID- 11733579 TI - Splenic T zone development is B cell dependent. AB - The factors regulating growth and patterning of the spleen are poorly defined. We demonstrate here that spleens from B cell-deficient mice have 10-fold reduced expression of the T zone chemokine, CCL21, a threefold reduction in T cell and dendritic cell (DC) numbers, and reduced expression of the T zone stromal marker, gp38. Using cell transfer and receptor blocking approaches, we provide evidence that B cells play a critical role in the early postnatal development of the splenic T zone. This process involves B cell expression of lymphotoxin (LT)alpha1beta2, a cytokine that is required for expression of CCL21 and gp38. Introduction of a B cell specific LTalpha transgene on to the LTalpha-deficient background restored splenic CCL21 and gp38 expression, DC numbers, and T zone size. This work also demonstrates that the role of B cells in T zone development is distinct from the effect of B cells on splenic T cell numbers, which does not require LTalpha1beta2. Therefore, B cells influence spleen T zone development by providing: (a) signals that promote T cell accumulation, and: (b) signals, including LTalpha1beta2, that promote stromal cell development and DC accumulation. Defects in these parameters may contribute to the immune defects associated with B cell deficiency in mice and humans. PMID- 11733580 TI - Sialylated O-glycans and sulfated tyrosines in the NH2-terminal domain of CC chemokine receptor 5 contribute to high affinity binding of chemokines. AB - The chemokine receptor CCR5 plays an important role in leukocyte chemotaxis and activation, and also acts as a coreceptor for human and simian immunodeficiency viruses (HIV-1, HIV-2, and SIV). We provide evidence that CCR5 is O-glycosylated on serine 6 in the NH2 terminus. The O-linked glycans, particularly sialic acid moieties, significantly contribute to binding of the chemokine ligands. By contrast, removal of O-linked oligosaccharide exerted little effect on HIV-1 infection. Sulfation of specific tyrosine residues in the CCR5 NH2 terminus was important for efficient beta-chemokine binding. Thus, as has been observed for the binding of selectins and their ligands, O-linked carbohydrates and tyrosine sulfates play major roles in promoting the interaction of chemokines with CCR5. The resulting flexible arrays of negative charges on the CCR5 surface may allow specific, high-affinity interactions with diverse chemokine ligands. Although this is the first example of O-linked oligosaccharides and tyrosine sulfates playing a role in chemokine binding, the high density of serines, threonines and tyrosines in the N-termini of many CC chemokine receptors suggests that these posttranslational modifications may commonly contribute to chemokine binding. PMID- 11733581 TI - Cytotoxic T lymphocyte antigen 4 and CD28 modulate cell surface raft expression in their regulation of T cell function. AB - Coreceptors CD28 and cytotoxic T lymphocyte antigen (CTLA)-4 have opposing effects on TcR/CD3 activation of T cells. While CD28 enhances and CTLA-4 inhibits activation, the underlying molecular basis of these effects has yet to be established. In this context, ganglioside and cholesterol enriched membrane microdomains (rafts, GEMs) serve as centers of signaling in T cells. Although CD28 can promote TcR/raft colocalization, evidence is lacking on whether the surface expression of membrane rafts can be targeted by CTLA-4 in its modulation of T cell responses. In this study, we demonstrate that both CD28 and CTLA-4 profoundly alter the surface expression of membrane rafts during T cell activation. While CD28 increased expression and the number of peripheral T cells induced to express surface rafts in response to TcR ligation, CTLA-4 potently inhibited both TcR and TcR x CD28 induced raft expression on the surface of T cells. Consistent with this, CD28 increased the presence of the linker of activated T cells (LAT) in purified membrane rafts, while CTLA-4 coligation effectively blocked this increase. Further, the reversal of the CTLA-4 block with CD3/CD28 ligation was accompanied by an increase in surface raft expression and associated LAT. Our observations demonstrate for the first time that CTLA-4 targets the release of rafts to the surface of T cells, and provides a mechanism for the opposing effects of CD28 and CTLA-4 on costimulation. PMID- 11733582 TI - Hepatocyte nuclear factor 1alpha controls the expression of terminal complement genes. AB - The terminal components of the complement system contribute to host defense by forming the multiprotein membrane attack complex (MAC) which is responsible for cell lysis and several noncytotoxic effects. Most of the complement proteins are synthesized in the liver, but the mechanisms controlling their tissue-specific expression have not been elucidated. In this study we show that mice lacking the hepatic transcription factor hepatocyte nuclear factor 1alpha (HNF1alpha) fail to transcribe C5 and C8A complement genes. In addition, mRNAs encoding for several other terminal complement components or subunits are expressed at lower levels, including C8beta, C8gamma, and C9. We next used a reconstitution assay involving human sera with selective complement deficiencies to assess mouse complement activity. Sera from HNF1alpha-deficient mice showed negligible hemolytic activity of both C5 and C8alpha-gamma subunits. The activity of C8beta was severely affected despite only a 50% reduction in C8beta mRNA levels in the liver. This is reminiscent of C8alpha-gamma-deficient patients who accumulate extremely low levels of the C8beta subunit. Our results demonstrate that HNF1alpha plays a key role in the expression of C5 and C8A genes, two terminal complement component genes that are essential for the assembly of MAC as a result of complement activation. PMID- 11733583 TI - Maturation of marginal zone and follicular B cells requires B cell activating factor of the tumor necrosis factor family and is independent of B cell maturation antigen. AB - B cells undergo a complex series of maturation and selection steps in the bone marrow and spleen during differentiation into mature immune effector cells. The tumor necrosis factor (TNF) family member B cell activating factor of the TNF family (BAFF) (BLyS/TALL-1) plays an important role in B cell homeostasis. BAFF and its close homologue a proliferation-inducing ligand (APRIL) have both been shown to interact with at least two receptors, B cell maturation antigen (BCMA) and transmembrane activator and cyclophilin ligand interactor (TACI), however their relative contribution in transducing BAFF signals in vivo remains unclear. To functionally inactivate both BAFF and APRIL, mice transgenic for a soluble form of TACI were generated. They display a developmental block of B cell maturation in the periphery, leading to a severe depletion of marginal zone and follicular B2 B cells, but not of peritoneal B1 B cells. In contrast, mice transgenic for a soluble form of BCMA, which binds APRIL, have no detectable B cell phenotype. This demonstrates a crucial role for BAFF in B cell maturation and strongly suggests that it signals via a BCMA-independent pathway and in an APRIL-dispensable way. PMID- 11733584 TI - More on BSE/vCJD. PMID- 11733585 TI - Is the National Health Service at the edge of chaos? AB - We used chaos and complexity theory to analyse waiting-list data (1998-2001) pertaining to over 20 000 National Health Service (NHS) patients from general surgical, orthopaedic and neurosurgical units across England. Plots of frequency versus quarter-to-quarter change in waiting times revealed a power relation which seems independent of surgical specialty and hospital location. One interpretation of these findings is that, for the period in question, the NHS was a system at the edge of chaos. This hypothesis might explain why waiting times have resisted attempts at shortening. PMID- 11733586 TI - Aiming at averages. PMID- 11733589 TI - Inappropriate admissions: thoughts of patients and referring doctors. AB - Research on inappropriate hospital admissions has tended to neglect the views of the referring doctors and the patients. In this study, the Appropriateness Evaluation Protocol was applied to a random sample of 102 emergency medical admissions. The patients and doctors were then presented with a list of possible alternatives to admission that might have been used at the point of referral. Case notes were available for 88 patients. As judged by these, 28% of admissions were inappropriate, the commonest reason being the potential for treatment or tests to have been performed as outpatient procedures; next commonest was the possibility of lower level care. The response rate to the questionnaires was about two-thirds, for both doctors and patients. Of the general practitioners and casualty doctors who responded, 60% specified alternatives to admission that they would have considered, and the equivalent figure for patients was 70%. For both groups the major preferences were same-day outpatient assessment and admission to a community hospital. Referring doctors and patients, in this survey, favoured alternatives to acute medical care in proportions much higher than that of supposedly inappropriate admission. PMID- 11733587 TI - Osteoporosis in men. PMID- 11733588 TI - Cerebral palsy--medicolegal aspects. PMID- 11733590 TI - Comparative audit: the trouble with POSSUM. AB - The Physiological and Operative Severity Score for the enUmeration of Mortality and Morbidity (POSSUM) has been proposed for use in comparative audit between surgeons and between hospitals. To assess its feasibility, POSSUM scoring was attempted on admission in all patients under the care of two consultant surgeons over a six-month period. Scores were awarded only if all investigations necessary for POSSUM were performed; investigations unnecessary for effective treatment were not performed. 815 patient discharges were recorded over the six-month period, with 521 patients undergoing operative procedures. Of those undergoing an operation, scores could be allocated in only 155 (30%). Scoring systems such as POSSUM are procedure-based, thereby excluding those who do not undergo an operation. However, most of our operative cases were also excluded. Full POSSUM scoring will often require additional investigations. POSSUM is unlikely to be of use in the wider setting of comparative audit. PMID- 11733591 TI - Bilateral osteonecrosis of the tarsus coincident with glioblastoma multiforme. PMID- 11733592 TI - Multiple squamous skin carcinomas following excess sunbed use. PMID- 11733593 TI - Severe hypernatraemia in obstructive uropathy. PMID- 11733594 TI - Testicular capillary haemangioma in a child. PMID- 11733595 TI - Massive haemoptysis 27 years after surgery for coarctation of the aorta. PMID- 11733596 TI - Cardiac intraventricular thrombus in protein C deficiency. PMID- 11733597 TI - Whipple's disease of the appendix. PMID- 11733598 TI - Robert Lee, the uterine nervous system and a wrangle at the Royal Society 1839 1849. PMID- 11733600 TI - Students sitting finals: ready to be house officers? PMID- 11733601 TI - Students sitting finals: ready to be house officers? PMID- 11733602 TI - Students sitting finals: ready to be house officers? PMID- 11733603 TI - Students sitting finals: ready to be house officers? PMID- 11733604 TI - Training for necropsy. PMID- 11733605 TI - Students sitting finals: ready to be house officers? PMID- 11733606 TI - Requests for vasectomy: counselling and consent. PMID- 11733607 TI - Nephrotoxicity in the elderly due to co-prescription of ACE inhibitors and NSAIDs. PMID- 11733608 TI - Amputations at the London Hospital, 1852-1857. PMID- 11733609 TI - Oesophageal cancer surgery. PMID- 11733610 TI - Attitudes to torture. PMID- 11733611 TI - Where shall we send our paper? PMID- 11733612 TI - Transition from paediatric clinic to the adult service. PMID- 11733613 TI - Pickles of Wensleydale. PMID- 11733614 TI - Cytochrome P450 metabolites of arachidonic acid: novel regulators of renal function. PMID- 11733615 TI - Cyclooxygenase-2 and atherosclerosis: friend or foe? PMID- 11733616 TI - Cancer-related anaemia requires higher doses of epoetin alfa than chronic renal failure replacement therapy. PMID- 11733617 TI - Skin changes in dialysis patients: a review. PMID- 11733618 TI - Continuous flow peritoneal dialysis (CFPD): a glimpse into the future. PMID- 11733619 TI - Glucotoxicity of the peritoneal membrane: the case for VEGF. PMID- 11733621 TI - Influence of hepatocyte growth factor, epidermal growth factor, and mycophenolic acid on endothelin-1 synthesis in human endothelial cells. AB - BACKGROUND: Endothelin-1 (ET-1) is a potent vasoconstrictive peptide which plays an important pathophysiological role in ischaemic renal failure and drug-induced renal injury such as cyclosporin A (CsA)- and tacrolimus-associated nephrotoxicity. In contrast, hepatocyte growth factor (HGF) and epidermal growth factor (EGF) seem to accelerate renal regeneration after ischaemic and drug induced renal injury. This study aimed to investigate the influence of HGF and EGF on ET-1 synthesis in cultured human umbilical vein endothelial cells (HUVEC) and renal artery endothelial cells (RAEC). In addition, we have investigated whether mycophenolic acid (MPA), a new immunosuppressive drug, which in contrast to CsA and tacrolimus lacks nephrotoxic side effects, modulates ET-1 synthesis in endothelial cells. METHODS: ET-1 release was measured with a specific enzyme linked immunosorbent assay. ET-1 mRNA expression was investigated by reverse transcription polymerase chain reaction. RESULTS: HGF and EGF (0.001-10 nM) exerted a significant concentration-dependent inhibitory effect on ET-1 release by HUVEC and RAEC (minimum 56.1+/-4.3% of control, n=6, mean+/-SE). The suppressive effect of HGF and EGF on ET-1 synthesis was dose-dependently antagonized by the tyrosine kinase inhibitors tyrphostin AG1478, lavendustin A and methyl 2,5-dihydroxycinnamate. Incubation of HUVEC and RAEC with MPA (2.5, 10, 25, and 50 microg/ml) for 3-5 h induced a significant reduction of ET-1 mRNA expression. After 48 h incubation with MPA (1-50 microg/ml) a significant decrease of ET-1 release and DNA content per culture well was observed, whereas ET-1 release referred to the DNA content in the corresponding culture well did not differ significantly from controls. CONCLUSIONS: The present findings demonstrate that HGF and EGF reduce ET-1 synthesis in endothelial cells via their receptor tyrosine kinase activity and suggest that the renoprotective effects of HGF and EGF might be linked to their inhibitory action on ET-1 synthesis. This study also provides evidence that, in contrast to CsA and tacrolimus, MPA does not stimulate ET-1 synthesis. This might explain the clinical observation that renal function often improves when CsA or tacrolimus is replaced by mycophenolate mofetil. PMID- 11733620 TI - Medullary nephrocalcinosis associated with long-term furosemide abuse in adults. AB - BACKGROUND: The use of furosemide is well recognized as a predisposing factor of nephrocalcinosis in infants. Although furosemide is widely used for various medical conditions in adults, its association with nephrocalcinosis in adults is not well established. METHODS: We studied 18 consecutive adult patients (male:female ratio 1:17, age range 21-59 years) who habitually took furosemide to control weight or oedema for long periods of time (range 3-25 years). The daily dose of continuous intake of furosemide ranged from 40 to 2800 mg. Nephrocalcinosis was evaluated using renal ultrasonography (US), computed tomography (CT), or kidney biopsies. RESULTS: Renal US and CT revealed bilateral nephrocalcinosis of the medullary pyramids in 15 (83.3%) out of 18 patients. The duration of furosemide abuse was similar between nephrocalcinosis positive (NC(+)) and nephrocalcinosis negative (NC(-)) groups. The daily dose of furosemide was nearly 10 times higher in the NC(+) group (range 120-2800 mg, mean 538 mg) than the NC(-) group (range 40-80 mg, mean 67 mg). All patients showed variable degrees of renal insufficiency and there was no difference in creatinine clearance between the NC(+) and NC(-) groups (P>0.05). Kidney biopsies performed in three patients showed focal tubulo-interstitial fibrosis and atrophy and calcifications were observed in outer medullary tubulo-interstitium. CONCLUSIONS: Long-term furosemide abuse can cause medullary nephrocalcinosis in adults, and the risk of developing of nephrocalcinosis seems to be correlated with the daily dose of furosemide. We suggest that long-term furosemide abuse should be suspected in adult patients when medullary nephrocalcinosis is incidentally detected by US or CT. PMID- 11733622 TI - No evidence for point mutations of the calcium-sensing receptor in familial idiopathic hypercalciuria. AB - BACKGROUND: Idiopathic hypercalciuria (IH) is frequently associated with nephrolithiasis. As 40% of patients have a positive familial history of IH, an autosomal dominant mode of inheritance has been suggested. Numerous genes have been studied in this regard but none have been found to be linked to IH. Mutation of the calcium-sensing receptor (CaR) has never been studied. Therefore, we conducted a study to detect such mutations. METHODS: Seven families with IH and nephrolithiasis were recruited in a prospective study. Forty-two family members underwent 24-h urine calcium measurement. Twenty-five of them with 24-h hypercalciuria also underwent extensive metabolic evaluation. Blood samples were collected in one or two affected family members in each family and exons 2-7 of the CaR gene were sequenced. RESULTS: In the seven families, at least one parent and more than half of the children had hypercalciuria (21/30), consistent with autosomal dominant inheritance. Among the nine affected family members whose CaR gene has been studied, all nine had absorptive hypercalciuria, three also had fasting hypercalciuria, and one had renal phosphorous leak. No mutation of the CaR gene was detected in these seven families. Two previously reported polymorphisms were detected, each of them in five families: A986S and C-to-T change at -60 in intron 5. CONCLUSION: In these seven families, IH is not related to the CaR gene mutation. Although we cannot exclude that point mutations can be found in other families, familial IH does not seem to be generally associated with CaR mutation. PMID- 11733623 TI - Angiotensin-converting enzyme activity and the ACE Alu polymorphism in autosomal dominant polycystic kidney disease. AB - BACKGROUND: Previous studies concerning Alu I/D polymorphism in the ACE gene and ADPKD severity have used the Alu genotypes as a representative of the true biological variable, namely ACE activity. However, wide individual and ethnic differences in the proportion of variance in ACE activity explained by the I/D genotype may have confounded these studies. This investigation examines the association between ADPKD severity and ACE in terms of plasma enzyme activity and I/D genotypes in individuals from three different countries. METHODS: Blood samples were collected from 307 ADPKD patients (116 Australian, 124 Bulgarian and 67 Polish) for determination of ACE activity levels and I/D genotypes. Chronic renal failure (CRF) was present in 117 patients and end-stage renal failure (ESRF) in 68 patients. RESULTS: ACE activity was related to the I/D genotype, showing a dosage effect of the D allele (P=0.006). The proportion of variance due to the Alu polymorphism was 14%. No difference in ACE activity and I/D genotype distribution was found between patients with CRF versus normal renal function (P=0.494; P=0.576) or between those with ESRF versus those without ESRF (P=0.872; P=0.825). No effect of the I/D genotype on age at development and progression to renal failure (CRF; ESRF) was detected in the overall group, and in subgroups based on ethnic origin, linkage status and sex. CONCLUSION: ACE is not likely to play a role as a determinant of ADPKD phenotype severity. PMID- 11733624 TI - Clinical and morphological features of kidney involvement in primary Sjogren's syndrome. AB - BACKGROUND: Primary Sjogren's syndrome is a connective tissue disorder affecting primarily the lacrimal and salivary glands, resulting in xerophtalmia and xerostomia. Extraglandular manifestations are frequent and may include renal involvement. METHODS: We studied the prevalence and nature of kidney involvement in 60 Italian patients with primary Sjogren's syndrome, diagnosed according to the European classification criteria. The following renal laboratory tests were performed in all patients: electrolytes in serum and in 24-h urine, creatinine in serum and in 24-h urine, venous pH and HCO(3)(-), urinalysis, urine culture, urinary osmolality and urine pH. A water deprivation test was performed in patients with morning urine osmolalities below the reference values adjusted for age. An oral ammonium chloride loading test was performed in patients with urine pH above 5.5 from morning samples. Renal biopsy was performed in patients with renal involvement. RESULTS: Sixteen patients (27%) had laboratory evidence of tubular and/or glomerular dysfunction. A variable degree of creatinine clearance reduction was found in eight patients (13%); frank distal tubular acidosis in three (5%); hypokalaemia in four (7%); and pathological proteinuria in 12 (20%). Urine concentrating capacity was defective in 10 out of 48 (21%) tested patients. Only four patients presented with overt clinical manifestations, including hypokalaemic tetraparesis (1), nephrotic syndrome (2), recurrent renal stones with flank pain and haematuria (1). In two patients, signs of renal involvement preceded the onset of sicca syndrome. Renal biopsies from nine patients showed tubulo-interstitial nephritis in six and glomerular disease in three. Patients with renal involvement had a significantly shorter disease duration compared with patients without renal abnormalities. CONCLUSIONS: Kidney involvement is a frequent extraglandular manifestation of primary Sjogren's syndrome. It is rarely overt and may precede the onset of subjective sicca syndrome. PMID- 11733625 TI - Renal function and structure in albuminuric type 2 diabetic patients without retinopathy. AB - BACKGROUND: In type 2 diabetic patients without retinopathy the cause of albuminuria is heterogeneous and our knowledge of the relationship between kidney structure and function in these patients is limited. Therefore, a long-term study evaluating the structural-functional relationship in albuminuric type 2 diabetic patients without retinopathy was performed. METHODS: Mesangial volume of total glomerular volume (Vv (mes/glom)), fractional area of focal interstitial fibrosis and tubular atrophy of cortical area (FF) and percentage of sclerosed glomeruli (S/G) were measured on kidney biopsies from 49 type 2 diabetic patients without retinopathy. Glomerular filtration rate (GFR) was determined at least 3 times (median 8 (range 3-20)) in each patient. Patients were followed for 7.0 (1.1-17) years. Albuminuria and blood pressure were measured every 3-6 months. RESULTS: Biopsies revealed diabetic glomerulopathy (DG-group) in 69% of the patients (27 male/7 female) and normal glomerular structure (n=9) or glomerulonephritis (n=6) were found in 31% (13 male/2 female) (NDG-group). In the DG-group GFR decreased from 97+/-5 to 66+/-5 ml/min/1.73 m(2) (mean+/-SE) (P<0.001), with a rate of decline in GFR of 5.3+/-0.8 ml/min/year and in the NDG-group from 93+/-7 to 74+/ 11 ml/min/1.73 m(2) (P<0.01), with a rate of decline in GFR of 3.2+/-0.9 ml/min/year, P=0.09 between groups. Mean arterial blood pressure decreased from 109+/-2 to 100+/-2 mm Hg (P<0.001) (DG-group) and remained unchanged in the NDG group. An association between Vv (mes/glom) and rate of decline in GFR was revealed mainly in the NDG-group (DG-group; r=0.31, P=0.07 and NDG-group; r=0.74, P<0.01). Furthermore, the rate of decline in GFR seemed to be associated with FF in the NDG group (r=0.48, P=0.07). Percentage of S/G was not associated with the rate of decline in GFR. Vv (mes/glom) was associated with mean albuminuria during follow-up in the DG group; r=0.38, P<0.03 (NDG group; r=0.51, P=0.09). Albuminuria was an independent predictor of the rate of decline in GFR in both groups (DG-group; r=0.40, P<0.05 and NDG-group; r=0.61, P<0.01). CONCLUSIONS: Our study revealed a tendency to a faster rate of decline in GFR in the DG-group compared to the much smaller NDG-group, characterized by marked heterogeneity of the underlying kidney lesions and rate of GFR loss. A large mesangial volume fraction was associated with increased albuminuria and loss in GFR. Albuminuria acted as a progression promoter in both groups. PMID- 11733626 TI - Candidaemia in patients with dialysis-dependent acute renal failure: aetiology, predisposing and prognostic factors. AB - BACKGROUND: Infections remain the major cause of death among patients with acute renal failure (ARF), especially in severe ARF necessitating dialysis therapy (ARF(d)). Although the clinical features and outcomes of candidaemia in various patient populations have been described, data concerning candidaemic episodes among patients with ARF(d) are scarce. This study investigated the aetiology, predisposing, and prognostic factors for candidaemia in the ARF(d) patient population. Three patient groups were investigated in this study. METHODS: During an 8-year study period from January 1992 to December 1999, 37 candidaemic episodes that developed among 653 ARF(d) patients were assigned to ARF(d) candidaemic group, and 170 candidaemic episodes developing in patients without ARF(d) or chronic uraemia as the non-ARF(d) candidaemic group, and 28 matched ARF(d) patients without candidaemia were assigned to the ARF(d) control group. Among these groups, clinical characteristics in ARF(d) candidaemia patients, predisposing factors, and outcomes were compared. Four management strategies including central catheter removal, anti-fungal therapy, both, or neither were applied. The prognostic factors for attributable death were evaluated by univariate analysis followed by the multivariate logistic regression analysis. RESULTS: The proportion of ARF(d) patients with candidaemia was significantly higher than in patients who had no ARF(d) or chronic uraemia (5.7% vs 0.15%, P<0.001). Compared with the non-ARF(d) candidaemic group, systemic lupus erythematosus (SLE), administration of corticosteroid, and central venous catheter-associated candidaemia were more common in the ARF(d) candidaemic group (P<0.05). In matched case-control study, multiple antibiotic usage was shown to be a predisposing factor for developing candidaemia in patients with ARF(d), and corticosteroid therapy has a marginal significance (P=0.059). The occurrence of candidaemia increased the mortality rate of ARF(d) (71% vs 39.2% in ARF(d) control group, P<0.05). By multivariate logistic analysis, the variables associated with attributable death in ARF(d) candidaemic group were identified to be an APACHE II score of >or=18, and anti-fungal therapy for >48 h. Central venous catheters were removed in 32 (86.5%) of the 37 ARF(d) candidaemic patients, among whom the 18 patients who had received anti-fungal therapy for >48 h had a lower attributable death rate than those patients who had not (27.8% vs 64.3%, P<0.05). Of the remaining five patients who did not have their catheter removed, three patients subsequently died and two patients improved only after catheter removal. CONCLUSIONS: The higher prevalence of candidaemia in ARF(d) patients is due to their underlying illnesses and multiplicity of predisposing factors, rather than ARF and dialysis therapy per se. Predisposing factors include SLE, indwelling central venous catheter, multiple antibiotic usage, and corticosteroid therapy. Prompt anti-fungal therapy and catheter removal should be mandatory for ARF(d) patients with candidaemia. PMID- 11733627 TI - Longer duration of predialysis nephrological care is associated with improved long-term survival of dialysis patients. AB - BACKGROUND: Late nephrological referral of chronic renal failure patients has been shown to be associated with high morbidity and short-term mortality on dialysis. However, the impact of predialysis nephrological care duration (PNCD) on the long-term survival of dialysis patients had not been evaluated. METHODS: We studied data from all 1057 consecutive patients who started dialysis treatment at the Necker Hospital from 1989 to 1998 (mean age at start of dialysis 53.8+/ 17.2 years (range 18-91 years), excluding from analysis patients who presented with acute renal failure (n=60) or advanced malignancy (n=35). We evaluated the effects of PNCD and clinical risk factors on all-cause mortality after long-term follow-up on dialysis. RESULTS: Among the 1057 patients analysed (13.2% diabetics), PNCD was <6 months in 258 patients, 6-35 months in 267 patients, 36 71 months in 227 patients and >or=72 months in 307 patients. Cardiovascular (CV) morbidity, namely a history of myocardial or cerebral infarction, peripheral arteriopathy, and/or cardiac failure, before starting dialysis was 39.6% and 37.4%, respectively, in patients followed for <6 months or 6-35 months, compared with 24.4% in those followed for 36-71 months and 19.9% in those followed for >or=72 months (P<0.001). Five-year survival was significantly lower in patients with a PNCD of <6 months (59+/-4.1%) than for 36-71 months or >or=72 months (77.1+/-3.7 and 73.3+/-3.6%, respectively, P<0.001), but similar to those followed for 6-35 months (65.3+/-3.9%, NS). By Cox proportional hazard analysis, PNCD <6 months, age, diabetes and prior CV disease were independent predictive factors of all-cause death on dialysis. CONCLUSIONS: This study provides suggestive evidence that longer duration of regular nephrological care in the predialysis period, at least for several years prior to the start of dialysis, is associated with a better long-term survival on dialysis. Such data strongly support the argument for early referral and regular nephrological care of chronic renal failure patients. PMID- 11733628 TI - Salvage of immature forearm fistulas for haemodialysis by interventional radiology. AB - BACKGROUND: The goal of this article is to assess the value of endovascular techniques for the salvage of fistulas that fail to mature. METHODS: Over a 6 year period, 52 dysfunctional and 17 thrombosed immature forearm fistulas (mean age 10 weeks) were treated by interventional radiology. Angiography was performed by puncture of the brachial artery but dilation of underlying stenoses was performed after cannulation of the fistula itself, whenever possible, with a balloon never smaller than 5 mm. Embolization or ligation of any type of vein was never indicated and never performed. For thrombosed fistulas, significant clots were removed by manual catheter-directed aspiration. A covered stent (Passager) was used in cases of dilation-induced rupture not controlled by balloon tamponade. RESULTS: An underlying stenosis was diagnosed in 100% of cases. Half of them were located in the anastomotic area. The initial success rate of interventional radiology was 97%. Dilation-induced rupture occurred in nine cases (13%) but stents were necessary in only two cases. The rate of significant clinical complications was 2.8% (bacteraemia, pseudoaneurysm). Primary and secondary patency rates at 1 year were 39 and 79%, respectively. CONCLUSIONS: Delayed maturation of native fistulas should lead systematically to imaging as an underlying stenosis is diagnosed in all cases. Interventional radiology can treat the majority of cases and achieve a 97% success rate but early recurrence of stenoses can occur. Multidisciplinary re-evaluation of the patient must, therefore, be performed after radiological salvage of the fistula. PMID- 11733629 TI - Independent predictors of restenosis after percutaneous coronary revascularization in haemodialysis patients. AB - BACKGROUND: Percutaneous balloon angioplasty has become a well-established and routine procedure for coronary revascularization of haemodialysis patients with coronary artery disease. However, the incidence of restenosis after balloon angioplasty is significantly higher in haemodialysis patients than in the general population. We performed a retrospective study comparing balloon angioplasty with coronary stenting in haemodialysis patients. We evaluated the long-term clinical and angiographic outcome after successful percutaneous coronary revascularization in haemodialysis patients. METHODS: A total of 103 consecutive haemodialysis patients (123 lesions) underwent procedurally and clinically successful percutaneous revascularization. Patients were treated with three different strategies: (i) balloon angioplasty in 55 patients (69 lesions); (ii) coronary stenting with balloon angioplasty in 23 patients (25 lesions); and (iii) coronary stenting with rotational atherectomy in 25 patients (29 lesions) who had severely calcified stenotic coronaries. RESULTS: The rates of in-hospital mortality were similar in the three groups. The 1-year incidence of overall events and major adverse cardiac events (MACE) were significantly higher in the balloon group than in the stent with/without rotational atherectomy groups (75% vs 36 and 28%, P<0.01; 71% vs 32 and 28%, P<0.01). Use of coronary stenting (relative risk=0.006, P<0.001) and the presence of calcified coronary lesion (relative risk=68.2, P<0.001) were independent predictors of the 1-year MACE-free survival after percutaneous revascularization. The 3-year MACE-free survival rate was significantly lower in the balloon group than in the stent with/without rotational atherectomy groups (11% vs 33 and 47%, P<0.005 and P<0.001). CONCLUSIONS: This study shows that coronary stenting reduces the incidence of MACE in haemodialysis patients with/without calcified coronary lesions. Moreover, coronary stenting reduces the restenosis rate of both complex and restenotic lesions, and rotational atherectomy prior to coronary stenting reduces the restenosis rate of the severely calcified coronary lesions. These results suggest that coronary stenting with/without rotational atherectomy has led to an improved long-term outcome in the haemodialysis patients with coronary artery disease. PMID- 11733630 TI - Comparison of body fluid distribution between chronic haemodialysis and peritoneal dialysis patients as assessed by biophysical and biochemical methods. AB - BACKGROUND: The control of extracellular volume is a key parameter for reducing hypertension and the incidence of cardiovascular mortality in dialysis patients. In recent years bioimpedance measurement (BIA) has been proven as a non-invasive and accurate method for measuring intracellular and extracellular fluid spaces in man. In addition, plasma atrial natriuretic peptide (ANP) and cyclic guanosine monophosphatase (cGMP) concentrations have been shown to reflect central venous filling. Using these methods, we compared body fluid status between stable patients on haemodialysis and peritoneal dialysis. METHODS: Thirty-nine chronic haemodialysis patients, 43 chronic peritoneal dialysis patients and 22 healthy controls were included in the study. Multifrequency BIA was performed using the Xitron BIS4000B device (frequencies from 5 to 500 kHz were scanned and fitted) in patients before and after haemodialysis. Peritoneal dialysis patients were measured after drainage of the dialysate. Plasma ANP and cGMP levels were measured in plasma using a (125)I solid phase RIA. Serum albumin concentrations and serum osmolality were measured in all patients. The body fluid data were analysed in relation with the clinical findings. RESULTS: Total body water (TBW) was 0.471+/-0.066 l/kg before haemodialysis and 0.466+/-0.054 l/kg after haemodialysis. Peritoneal dialysis patients had a TBW (0.498+/-0.063 l/kg) that was greater than the before and after dialysis values of haemodialysis patients. The extracellular body fluid (V(ecf)) was increased pre-haemodialysis. It was even greater in peritoneal dialysis patients compared with patients both pre- and post-haemodialysis (pre 0.276+/-0.037 l/kg; post 0.254+/-0.034 l/kg; peritoneal dialysis 0.293+/-0.042 l/kg, P<0.05). However, plasma ANP concentrations (representing intravascular filling) in peritoneal dialysis patients were comparable with post-haemodialysis values (284+/-191 pg/ml vs 286+/-144 pg/ml). The correlation coefficient between sysRR and V(ecf) was r=0.257 in haemodialysis (P=0.057) and r=0.258 in peritoneal dialysis (P<0.05). A significant negative correlation was found between serum albumin and V(ecf)/TBW in peritoneal dialysis patients (r= -0.624). CONCLUSION: Body fluid analysis by BIA demonstrated that TBW and V(ecf) were increased in peritoneal dialysis patients, and were comparable or even greater than values found before haemodialysis. However, plasma ANP levels indicated that intravascular filling was not increased in peritoneal dialysis. The ratio of V(ecf) to TBW was correlated to systolic pressure and negatively to serum albumin in peritoneal dialysis patients. PMID- 11733631 TI - Body mass index and mortality in 'healthier' as compared with 'sicker' haemodialysis patients: results from the Dialysis Outcomes and Practice Patterns Study (DOPPS). AB - BACKGROUND: Haemodialysis (HD) patients with lower body mass index (BMI) have a higher relative mortality risk (RR), irrespective of race. However, only Asian Americans treated with HD have been found to have an elevated RR with higher BMI. Asian Americans on HD are 'healthier' than other race groups (i.e. have better overall survival). We hypothesized that an increased mortality risk might be associated with high BMI in a variety of other 'healthier' subgroups of HD patients. METHODS: The prospective Dialysis Outcomes and Practice Patterns Study (DOPPS) provided baseline demographic, comorbidity and BMI data on 9714 HD patients in the US and Europe (France, Germany, Italy, Spain, and the UK) from 1996-2000. Using multivariate survival analyses, we evaluated BMI-mortality relationships in HD subpopulations defined by continent, race (black and white), gender, tertiles of severity of illness (based on a score derived from comorbid conditions and serum albumin concentration), age (<45, 45-64, >or=65), smoking, and diabetic status. RESULTS: Relative mortality risk decreased with increasing BMI. This was statistically significant (P<0.007) except for the smallest subgroup of patients who were <45 years old and were also in the healthiest tertile of comorbidity. All else equal, BMI <20 was consistently associated with the highest relative mortality risk. Overall a lower relative mortality risk (RR) as compared with BMI 23-24.9, was found for overweight (BMI 25-29.9; RR 0.84, P=0.008), for mild obesity (BMI 30-34.9; RR 0.73, P=0.0003), and for moderate obesity (BMI 35-39.9; RR 0.76, P=0.02). CONCLUSION: In a wide variety of HD patient subgroups, differing with respect to their baseline health status, increasing body size correlates with a decreased mortality risk. This contrasts with the association between BMI and mortality in the general population, and deserves further study. PMID- 11733632 TI - An African community-based chronic ambulatory peritoneal dialysis programme. AB - BACKGROUND: The reasons for failure of continuous ambulatory peritoneal dialysis (CAPD) are varied. Against a background of mass poverty, poor resources, and the cheaper cost of CAPD it is the primary choice of dialysis. The aims of this study were to determine infection rates and document factors responsible for CAPD failure. METHODS: We report a prospective study in a large African tertiary hospital and its community based satellite clinics. Infection rates as well as factors that may influence them were studied. Sites of infections were documented and causes of CAPD failure recorded. All patients qualifying for dialysis from January 1998 to July 1999 were included. RESULTS: Eighty-four patients were enrolled. There were 55 males and 29 females. The mean age was 39+/-10 (range 16 71) years and mean duration on dialysis at the end of the trial period was 17 months. The peritonitis rate was one episode every 27.9 patient months. Attrition to haemodialysis occurred in 16.6% of patients (n=14) and loss to follow-up in 29.8% (n=25). Fourteen patients regained renal function or were transplanted. Peritonitis appeared to be related to a poor BAD-C score (Bara Adapted Dialysis Compliance), i.e. combination of clinical status and clinic visits (P=0.07). The odds ratio for failure of CAPD with peritonitis was 5.3 times higher (confidence interval (CI) 1.7-17.1; P=0.0085). A low BAD-C score was a significant indicator of CAPD 'failure' (P=0.0001). The natural turnover rate of patients was 46%. Home conditions, employment, and education levels did not correlate with CAPD 'failure'. CONCLUSION: The peritonitis rate and aetiology are similar to the developed world. Socioeconomic factors did not appear to play a role in peritonitis rates or CAPD failure. PMID- 11733633 TI - Chronic allograft nephropathy--biopsy findings and outcome. AB - BACKGROUND: Chronic allograft nephropathy (CAN) is a composite term for various types of damage to a kidney transplant. We wanted to analyse its components in relation to baseline biopsy findings, transplant function, and outcome. METHODS: Among renal transplantations performed from 1985 to 1997, 156 were identified where allograft biopsies had been obtained on clinical indication 6 months after transplantation or later, baseline biopsies were available in each case and the patient's original disease was known. Time after transplantation was median 2.2 years (range 0.5-13). The biopsies were reviewed and the Banff 1997 CAN score obtained. RESULTS: All but one late biopsy showed some CAN grade, 48% grade II, and 7.5% grade III. Acute tubulointerstitial rejection was seen in 9% but vascular rejection in only 3%. Arterial wall thickening was present in 66% of the late biopsies, correlated with donor age and its presence at baseline but also with time after transplantation. The Banff CAN score and serum creatinine level were both independent predictors of further graft survival, relative risk 0.35 (confidence interval 0.15-0.82, P=0.015) for CAN grade I vs III and 0.30 (0.14 0.67, P=0.003) for serum creatinine <170 vs >250 micromol/l. Presence of arterial wall thickening had no prognostic impact. CONCLUSION: The CAN grade is predictive of further graft survival independently of the serum creatinine level. Interstitial fibrosis and tubular atrophy are more prominent features of chronic graft damage than vascular rejection. Unspecific arterial wall thickening is partly dependent on baseline conditions and lacks prognostic impact in this late stage. PMID- 11733634 TI - Oxalate removal by daily dialysis in a patient with primary hyperoxaluria type 1. AB - BACKGROUND: Dialysis patients with primary hyperoxaluria are exposed to risks and hazards associated with calcium oxalate salt deposition in body tissues, since regular dialysis treatment does not adequately correct hyperoxalaemia. The purpose of this study was to evaluate oxalate mass removal using various dialysis modes in a patient suffering from primary hyperoxaluria type 1 (PH1). METHODS: Oxalate kinetics during daily haemodialysis was compared with that of standard haemodialysis (STD HD) and haemodiafiltration (HDF) using high flux dialysers (FB 170 H and FB 210 U, Transdial, Paris, France). All dialysis sessions lasted for 4 h. Blood was withdrawn and spent dialysate was collected in plastic bags every hour to evaluate mass removal. Oxalate concentration in plasma and in spent dialysate was determined by an enzymatic method. Oxalate generation, distribution volume and tissue deposition were calculated using single-pool models adapted from previous studies. RESULTS: Although no significant difference was found in mass removal per session between dialysis strategies and dialyser types, weekly mass removal with daily HD was about 2 times greater than with STD HD or HDF. Even when daily HD was performed, the oxalate generation rate-mass removal ratio (G/R ratio) remained at a value of approximately 2. CONCLUSION: Although daily HD sessions led to a substantial increase in weekly oxalate removal, all three types of renal replacement therapy were insufficient to compensate for estimated oxalate generation. To eliminate sufficient amounts of oxalate generated in PH1 patients, at least 8 h of daily dialysis with a high-flux membrane would probably be required. Renal replacement therapy for PH1 patients needs be improved further. PMID- 11733635 TI - Late onset of renal disease in nephronophthisis with features of Joubert syndrome type B. PMID- 11733636 TI - Severe metabolic acidosis during haemodialysis: a rare but life threatening complication. PMID- 11733637 TI - Massive rhabdomyolysis and life threatening hyperkalaemia in a patient with the combination of cerivastatin and gemfibrozil. PMID- 11733638 TI - Fibrosing cholestatic hepatitis possibly related to persistent parvovirus B19 infection in a renal transplant recipient. PMID- 11733639 TI - A patient with evidence of two underlying diseases causing hypercalcaemia. PMID- 11733640 TI - Two haemodialysis patients with unclear abdominal symptoms of similar origin. PMID- 11733641 TI - Recurrent Escherichia coli bacteraemia in a patient with chronic renal failure. PMID- 11733642 TI - Exercise-induced anuria in a renal allograft recipient. PMID- 11733643 TI - Destruction of a cervical vertebra in a patient with end-stage renal failure. PMID- 11733645 TI - Is the use of subcutaneous epoetin beta once a week really equivalent to a more frequent administration of the same total dose? PMID- 11733646 TI - Is it possible for studies comparing Y-set, double-bag, and standard systems of CAPD to be blinded? PMID- 11733647 TI - Henoch-Schonlein purpura nephritis associated with coagulase-negative staphylococci sepsis in a patient with myeloma. PMID- 11733648 TI - Urinary transforming growth factor-beta in patients with diabetic nephropathy: implications for the pathogenesis of tubulointerstitial pathology. PMID- 11733649 TI - Does long-term administration of sodium thiosulphate inhibit progression to renal failure in nephrocalcinosis? PMID- 11733650 TI - Homocysteine plasma levels correlate with intimal carotid artery thickness in haemodialysis patients. PMID- 11733651 TI - Flow measurement in dialysis shunts. PMID- 11733652 TI - Delayed splenic rupture as a cause of haemoperitoneum in a CAPD patient with amyloidosis. PMID- 11733654 TI - Identification of missense mutation (G365R) of the butyrylcholinesterase (BCHE) gene in a Japanese patient with familial cholinesterasemia. AB - A point mutation which caused a silent phenotype of human serum butyrylcholinesterase (BChE) was identified in the genomic DNA of a 57-year-old Japanese woman who visited our hospital because of pneumonia. The propositus exhibited an unusually low level of BChE activity, whereas her son and daughter had an intermediate level. Immunologically, there was an absence of BChE protein in the propositus's serum. DNA sequence analysis of the propositus demonstrated a point mutation at codon 365 (GGA-CGA), resulting in a Gly-Arg substitution. A family study showed her son and daughter to have the same mutation. PMID- 11733655 TI - Use of chromogenic tube and methyl blue-sabouraud agar for the identification of Candida albicans strains. AB - This study was performed to investigate the use of chromogenic tube and methyl blue-Sabouraud agar for the presumptive identification of Candida albicans. 124 clinical isolates, including 111 C.albicans and 13 Candida spp strains, which had been identified by morphology on cornmeal tween 80 agar and Vitek automated identification system, were included. Three different identification procedures, a) germ tube test, b) chromogenic tube test by using CHROMagar Candida and c) methyl blue-Sabouraud agar test, were performed to the strains. 88 of 111 (79.3%) C.albicans strains were detected to be positive by germ tube test. 87 (78.4%), 97 (87.4%) and 102 (91.9%) of these isolates were identified as C.albicans by chromogenic tube test after 2, 8 and 24 hours of incubation, respectively. 88 (79.3%), 92 (82.9%) and 88 (79.3%) of the isolates were correctly identified as C.albicans by methyl blue-Sabouraud agar test after 2, 8 and 24 hours of incubation, respectively. The sensitivity and specificity values were found to be 79.3 and 69.2 for the germ tube test. These values ranged between 78.4-91.9% and 69.2-76.9% for chromogenic tube test and 79.3-82.9% and 76.9-84.6% for methyl blue-Sabouraud agar depending on the incubation period. It can be concluded that the use of chromogenic tube and methyl blue-Sabouraud agar are rapid, simple and objective methods for the identification of C.albicans strains. PMID- 11733656 TI - Intraarterial infusion of papaverine and change of cerebral hemodynamics in symptomatic cerebral vasospasm. AB - In 43 cases with symptomatic cerebral vasospasm after aneurysmal subarachnoid hemorrhage treated by intraarterial infusion of papaverine (IAP), we studied cerebral hemodynamics by measuring cerebral circulation time (CCT) using digital subtraction angiogram. CCT on the middle cerebral artery site was defined as CCT LAV and on the anterior cerebral artery side as CCT-MAV. In the CCT-LAV, two phases were further defined; the arterial phase (CCT-A) and the capillary phase (CCT-CAP). Mean CCT-LAV before and after IAP was 6.35+/-1.69 sec, 4.91+/-1.56 sec, and mean CCT-MAV was 6.15+/-1.68 sec, 4.80+/-1.58 sec, each showing a significant shortening. Mean CCT-A before and after IAP was 0.274+/-0.105 sec, 0.226+/-0.066 sec and mean CCT-CAP was 6.00+/-1.62 sec and 4.60+/-1.55 sec. The shortening rate of CCT-A and CCT-CAP were 11.2+/-25.7% and 22.7+/-14.6% respectively. Our study confirmed that IAP shortened CCT and improved cerebral hemodynamics. Compared with CCT-A, CCT-CAP was shortened significantly, suggesting that IAP is working not only in the proximal vessels but also in the distal vessels, that is, effective for resolution of vasospasm in the view of cerebral microcirculation. Clinical outcome was not improved statistically, however, it is the fact that there are not a few cases improved neurologically and IAP is still useful for vasospasm as a means to directly dilate intracranial peripheral arteries. PMID- 11733657 TI - Flk-1 specific kinase inhibitor (SU5416) inhibited the growth of GS-9L glioma in rat brain and prolonged the survival. AB - BACKGROUND: Accumulating evidences suggest that tumor growth and metastasis depend on angiogenesis. At present, plenty of efforts are made to discover a chemical compound that specifically inhibits tumor angiogenesis either by reducing pro-angiogenic factor or increasing anti-angiogenic factors. OBJECT: SU5416, a novel, synthetic, potential inhibitor of angiogenesis specifically blocks the Flk-1/KDR tyrosine kinase activity. In vivo effect of SU5416 in the treatment of intracranial tumors has not been previously described. METHODS: We transplanted GS-9L cells into the right caudate nucleus of male Fisher 344 rats and administrated SU5416 intraperitoneally (i.p.) to investigate the impact of SU5416 on tumor angiogenesis and growth in vivo. Starting on Day 1 or Day 8, forty-two animals were treated with SU5416 at three different doses (e.g. 12.5, 25.0 and 50.0 mg/kg body weight) via i.p. injection every day until the end point. As a control, seven animals received no treatment and after implant fourteen animals were treated with vehicle (DMSO) only. RESULTS: SU5416 prolonged the survival in the treated groups without any significant systemic adverse effect. Median survival in the treated group started on Day.1 was statistically longer compared to that in the control groups (p<0.01). Histological analysis of the treated tumors showed an increase in necroses and reduced in vascularity compared to the control tumors. Furthermore, the number of apoptotic cells increased in the treated tumors on a TUNEL stain. CONCLUSION: Small molecular compounds, such as SU5416 may be useful therapeutics that specifically inhibits the enzymatic activity of Flk-1 kinase and downstream events of tumor angiogenesis. PMID- 11733658 TI - The amygdala and cardiovascular control. PMID- 11733659 TI - Subanesthetic concentration of sevoflurane increases regional cerebral blood flow more, but regional cerebral blood volume less, than subanesthetic concentration of isoflurane in human volunteers. AB - Both sevoflurane and isoflurane are used in moderate concentrations in neuroanesthesia practice. The limiting factors for using higher concentrations of inhalational anesthetics in patients undergoing neurosurgery are the agents' effects on cerebral blood flow (CBF) and cerebral blood volume (CBV). In particular, an increase in CBV, which is a key determinant of intracranial pressure, may add to the neurosurgical patient's perioperative risk. To compare the effects of a subanesthetic concentration (0.4 minimum alveolar concentration) of sevoflurane or isoflurane on regional CBF (rCBF), regional CBV (rCBV) and regional mean transit time (rMTT), contrast-enhanced magnetic resonance imaging perfusion measurements were made in spontaneously breathing human volunteers. Absolute changes in rCBF, regional CBV, and rMTT during administration of either drug in regions of interest outlined bilaterally in white and grey matter were nonparametrically (Mann-Whitney test) analyzed. Sevoflurane increased rCBF in practically all regions (absolute change, 4.44 +/- 2.87 to 61.54 +/- 2.39 mL/100g per minute) more than isoflurane did (absolute change, 12.91 +/- 2.52 to 52.67 +/ 3.32 mL/100g per minute), which decreased frontal, parietal, and white matter rCBF (absolute change, -1.12 +/- 0.59 to -14.69 +/- 3.03 mL/100g per minute). Regional CBV was higher in most regions during isoflurane administration (absolute change, 0.75 +/- 0.03 to 4.92 +/- 0.16 mL/100g) than during sevoflurane administration (absolute change, 0.05 +/- 0.14 to 3.57 +/- 0.14 mL/100g). Regional mean transit time was decreased by sevoflurane (absolute change, -0.18 +/- 0.05 to -0.60 +/- 0.04 s) but increased by isoflurane (absolute change, 0.19 +/- 0.03 to 0.69 +/- 0.04 s). In summary, regional CBV was significantly lower during sevoflurane than during isoflurane administration, although sevoflurane increased rCBF more than isoflurane, which even decreased rCBF in some regions. For sevoflurane and, even more pronouncedly, for isoflurane, the observed changes in cerebral hemodynamics cannot be explained by vasodilatation alone. PMID- 11733660 TI - Inhalation versus total intravenous anesthesia for lumbar disc herniation: comparison of hemodynamic effects, recovery characteristics, and cost. AB - The clinical effects, recovery characteristics, and costs of total intravenous anesthesia (TIVA), sevoflurane, and isoflurane anesthesia have been measured in various out-patient operations, but have not been evaluated in patients undergoing laminectomy or discectomy. In the current study, the authors assessed the hemodynamic characteristics, recovery, and cost analyzes after laminectomy and discectomy operations, comparing TIVA, sevoflurane, and isoflurane anesthesia. Sixty American Society of Anesthesiologists I and II patients were randomly divided into three groups, each consisting of 20 patients. Group I received propofol-alfentanil, Group 2 received sevoflurane-N2O, and Group 3 received isoflurane-N2O. At the end of surgery, the anesthetics were discontinued, and recovery from anesthesia was assessed by measuring the time until spontaneous eye opening and the time until response to verbal commands. The drug and delivery costs were calculated in United States dollars. No significant differences were found in the demographic data. Heart rate and mean arterial pressure decreased significantly after induction of anesthesia in the TIVA group, compared to the two other groups ( P < .05 for both comparisons). The fastest recovery was seen in the TIVA group. Incidences of postoperative nausea, vomiting, and pain were significantly reduced after TIVA ( P < .05 for both comparisons). Thus, TIVA patients required fewer additional drugs and showed the lowest additional costs in the post-anesthesia care unit. However, the total cost was significantly higher in the TIVA group than in the sevoflurane and isoflurane groups (52.73 dollars, 29.99 dollars, and 24.14 dollars, respectively) ( P < .05). Total intravenous anesthesia was associated with the highest intraoperative cost but provided the most rapid recovery from anesthesia, and the least frequent postoperative side effects. PMID- 11733661 TI - Brain compared to heart tissue oxygen pressure during changes in arterial carbon dioxide in the dog. AB - Myocardial tissue oxygen pressure (PmO2 ) and left anterior descending (LAD) artery blood flow were measured in dogs anesthetized with 1.5% isoflurane, and were then compared to brain tissue oxygen pressure (PbO2 ) and middle cerebral artery (MCA) blood flow during normocapnia, hypocapnia, and hypercapnia. A craniotomy was performed and a tissue probe (Codman, Inc.) that measures PO2, PCO2, and pH was inserted into the brain cortex in the MCA region (n = 8). Separately, after a thoracotomy, a probe was inserted into the middle myocardium of the left ventricle, within the distribution of the LAD, in eight dogs. Blood flow probes were placed on the LAD or MCA. Blood flow and tissue gases were measured during normocapnia (PaCO2 = 38 mm Hg), hypocapnia (PaCO2 = 26 mm Hg), and hypercapnia (PaCO2 = 53 mm Hg). Mean arterial pressure, heart rate, arterial gases, and pH were not different between brain and heart measurements. PbO2 was 21 +/- 9 mm Hg (mean +/- SD ), 40 +/- 16 mm Hg, and 47 +/- 11 mm Hg. PmO2 was 35 +/- 12 mm Hg, 40 +/- 14 mm Hg, and 48 +/- 15 mm Hg during hypocapnia, normocapnia, and hypercapnia respectively. During hypercapnia, LAD and MCA flow increased 50% and tissue oxygenation increased 20% ( P < .05). During hypocapnia, MCA flow and PbO2 decreased 50% ( P < .05), but LAD flow and PmO2 did not significantly change. These results indicated that LAD flow and myocardial PO2 were less responsive to hypocapnia than MCA flow and PbO2. PMID- 11733662 TI - Acute hypotension and bradycardia by medulla oblongata compression in spinal surgery. AB - A 71-year-old man was admitted to the hospital with subarachnoid hemorrhage caused by a cervical dural arteriovenous shunt. During surgery, the patient developed acute hypotension and bradycardia, probably caused by surgical compression of the medulla oblongata. During posterior fossa and upper cervical surgery, monitoring cardiovascular, respiratory, and evoked potential parameters is advocated. In the current case, only cardiovascular monitoring detected alteration of brain stem function. Anesthesiologists should be aware that surgical manipulation of the dorsal medulla might cause hemodynamic changes and expose patients to danger. Through close cardiovascular monitoring we can rapidly detect changes in vital signs, which allows prompt intervention to prevent irreversible neurologic deficits and potentially catastrophic patient outcome. PMID- 11733663 TI - Neurologic deficits after cervical laminectomy in the prone position. AB - New neurologic deficits are known to occur after spine surgery. We present four patients with cervical myeloradiculopathy who underwent cervical laminectomy, fusion, or both in the prone position, supported by chest rolls. Three patients were intubated and positioned while awake, whereas the fourth patient was positioned after induction. Surgeries were successfully carried out, except for transient episodes of relative hypotension intraoperatively. On recovery from anesthesia, all patients were noted to have new neurologic deficits. Immediate CT myelography or surgical reexploration was unremarkable. All patients improved gradually with administration of high-dose steroids and induction of hypertension. Use of the prone position with abdominal compression may compromise spinal cord perfusion and lead to spinal cord ischemia. The use of frames that prevent abdominal compression, as well as avoidance of perioperative arterial hypotension, is important in maintaining adequate spinal cord perfusion during and after decompressive spinal cord surgery. PMID- 11733664 TI - Dexmedetomidine infusion and somatosensory evoked potentials. AB - Intraoperative neurophysiologic monitoring requires information on the effects of anesthetic drugs because these drugs can directly alter evoked potentials, thus interfering with monitoring. We report on our evaluation of the effect of the recently introduced alpha2-adrenergic agonist, dexmedetomidine, on the somatosensory evoked potentials in two patients undergoing cervico-occipital fusion. Our results suggest that, although dexmedetomidine can affect the later cortical peaks of somatosensory evoked potentials (SSEPs), consistent and reproducible potentials can be recorded. PMID- 11733665 TI - Recovery after high-dose methylprednisolone and delayed evacuation: a case of spinal epidural hematoma. AB - Spinal epidural hematoma (SEH) is rare and not without serious sequelae. We report a patient who developed Brown-Sequard syndrome from SEH after fluoroscopic guided cervical steroid injection and favorable response to methylprednisolone (MP). A 56-year-old man reported immediate sharp shooting pain to the upper extremities on introduction of epidural toughy needle. A total of 5 mL of 0.2% ropivacaine and 120 mg methylprednisolone acetate suspension was administered at the C6-7 interspace. Within half an hour, a neurologic deficit occurred at C7-8 and right Brown-Sequard syndrome developed. Once SEH was suspected (3 hours after onset of neurologic deficit), a protocol of high-dose MP intravenous infusion was initiated. Immediate incomplete recovery of motor, sensory, and sphincteric functions was noted within 30 minutes of infusion. Emergency spinal C6-T2 bilateral decompressive laminectomies and evacuation SEH were performed within an expected delay (10 hours from the onset of neurologic deficit). Fluoroscopic guidance does not take the place of adherence to meticulous technique. An unexplained neurologic deficit after invasive spinal procedures should raise the concern for SEH. Early recognition and emergent evacuation remain the mainstay management for SEH. This case suggests some neuroprotection from MP in cases of cervicothoracic cord compression secondary to traumatic SEH. When potential risks for SEH exist, it is advisable not to administer local anesthetic so as not to interfere with neurologic assessment and delaying the diagnosis. PMID- 11733667 TI - Effects of sevoflurane and isoflurane on electrocorticographic activities in patients with temporal lobe epilepsy. AB - To compare the neuroexcitatory effects of sevoflurane and isoflurane, we recorded electrocorticograms (ECoG) during wakefulness and during sevoflurane and isoflurane anesthesia in six patients with temporal lobe epilepsy (TLE). These patients had subdural grid electrodes chronically implanted in the temporal region. During sevoflurane anesthesia at 1.5 minimum alveolar concentration (MAC) of the combination with 67% nitrous oxide (N2O), a marked increase in interictal paroxysmal activities was observed in four patients. Two patients had a slight increase in paroxysmal activities. Activated areas were widely distributed, not being confined to the ictal onset zone of spontaneous seizures. However, isoflurane anesthesia at 1.5 MAC of the combination with 67% N2O was associated with less increased paroxysmal activity. While the neuroexcitatory properties of sevoflurane proved greater than those of isoflurane, the widespread irritative response to sevoflurane administration was not helpful in localizing the epileptogenic area. PMID- 11733666 TI - Severe bradycardia during epilepsy surgery. AB - Several kinds of arrhythmia are known to occur during epileptic seizure, and bradycardia has been reported in patients with temporal lobe epilepsy. The authors review the anesthesia records of patients with intractable epilepsy. Forty-two consecutive patients with intractable epilepsy who underwent epilepsy surgery were examined. Anterior temporal lobectomy was performed on 29 patients, frontal lobectomy on 2 patients, and a subdural electrode was set on 11 patients. Anesthesia was induced with propofol, fentanyl, and vecuronium and maintained with sevoflurane-fentanyl, propofol-fentanyl, or fentanyl-droperidol. Severe bradycardia (13-39 beats/min) was seen in six patients. All six patients recovered within 1 minute of interrupting the surgical procedure and administering intravenous atropine, and the surgeries were completed with no complications. The authors believe the six events were sinus bradycardias. They all occurred during amygdalo-hippocampectomy in cases of temporal lobectomy. This type of hemodynamic change was not seen in any of the patients undergoing temporal lobectomy without hippocampectomy, in patients undergoing frontal lobectomy, or when setting subdural electrodes. Experimentally, it has been shown that stimulation of the limbic system, such as the hippocampus, amygdala, and insular cortex, induces bradycardia and hypotension resulting from increased parasympathetic flow via the vagus nerve. Severe bradycardia may thus occur during surgery for temporal lobe epilepsy, and hemodynamic changes should be watched carefully during amygdalo-hippocampectomy. PMID- 11733668 TI - Routine use of adjusted low-dose oral anticoagulants during the first three postoperative months after hip fracture in patients without comorbidity factors. AB - OBJECTIVE: This study reports the efficacy of adjusted low-dose oral anticoagulants in the prevention of fatal pulmonary embolism after hip fracture. STUDY DESIGN: Consecutive clinical study. PATIENTS: Five hundred patients with hip fracture received oral anticoagulants as exclusive treatment. These patients were considered preoperatively not to have an increased risk for the development of postoperative pulmonary embolism; they had no poorly controlled systemic illness and were followed-up for at least three months. The average age was seventy-four years (range, 65-80). Anticoagulant therapy was begun on the evening of the operation and continued for three months. The dose was adjusted to keep the prothrombin time between seventeen and twenty seconds. Neither phlebography, sonography, nor lung scan was done routinely. RESULTS: Among the 500 patients, 476 continued to take low-dose oral anticoagulant for three months after the operation. Eight pulmonary embolisms were observed, but none was fatal. Only one patient developed a postoperative hematoma requiring surgical reoperation. CONCLUSIONS: In the absence of poorly controlled systemic illness, low-dose oral anticoagulants can be used after hip fracture. PMID- 11733669 TI - Outcome of plate fixation of olecranon fractures. AB - OBJECTIVES: To evaluate the functional outcome of plate fixation for displaced olecranon fractures, both simple and comminuted. DESIGN: Retrospective patient, chart, and radiographic review. SETTING: Academic teaching hospital. PATIENTS: Twenty-five patients who underwent plate fixation of displaced olecranon fractures by two surgeons were independently reviewed at an average follow-up of thirty-four months (range, 15-69 months). MAIN OUTCOME MEASUREMENTS: Physical capability was assessed by measuring range of motion and isometric elbow strength. Patient-rated outcomes were evaluated using the SF-36, DASH, Mayo Elbow Performance Index (MEPI), and visual analogue scales for patient satisfaction and pain. Radiographs were evaluated preoperatively, postoperatively, and at the time of final review. RESULTS: The average patient age was fifty-four years (range, 14 81 years). The Mayo classification of fractures was fourteen Type II and eleven Type III. An adequate reduction was maintained in all elbows until union. Physical capability measures indicated nonsignificant side-to-side differences in motion or strength, except for supination motion, which was reduced in the injured arm (p = 0.003). The MEPI-rated outcome was twenty-two excellent or good. Patient satisfaction was high (9.7/10), with a low pain rating (1/10). The mean DASH score was consistent with almost normal upper extremity function. The SF-36 showed no difference in physical health as compared with the average American population. Twenty percent of patients required plate removal because of prominence of the internal fixation. The outcome was not influenced by fracture pattern. CONCLUSION: Plate fixation is an effective treatment option for displaced olecranon fractures with a good functional outcome and a low incidence of complications. PMID- 11733670 TI - Concurrent dorsal dislocations and fracture-dislocations of the index, long, ring, and small (second to fifth) carpometacarpal joints. AB - OBJECTIVE: To review the outcome of patients with concurrent dorsal dislocations and fracture-dislocations of the second, third, fourth, and fifth carpometacarpal (CMC) joints treated with open reduction and internal fixation (ORIF). DESIGN: Retrospective review. SETTING: Level 1 trauma center. PATIENTS: Between 1991 and 1997, twelve multiply injured patients with the described CMC injury complex (one open injury) were treated with ORIF (eleven patients) or percutaneous wire fixation (one patient) by the same surgeon. INTERVENTION: Treatment consisted of ORIF with Kirschner wires followed by splints and immediate metacarpophalangeal and interphalangeal joint range of motion exercises. MAIN OUTCOME MEASUREMENTS: Grip strength, wrist and finger range of motion, pain, need for additional surgery, and return to work. RESULTS: Of the ten patients available for follow-up (mean, three years), all had been treated with ORIF (eight within forty-eight hours of injury and two had treatment delayed for four weeks because of delayed diagnosis and management of more serious injuries). Three patients had additional surgery (planned secondary second and third CMC arthrodeses). Grip strength of the operated hand in the five patients with JAMAR testing was 50 percent (n = 3), 75 percent (n = 1), and 90 percent (n = 1) of that in their contralateral hands. Five patients were pain-free, and five reported occasional, activity-related pain. The five patients who worked before the injury returned to their previous occupations (one with slightly modified duties). CONCLUSION: This is the largest series of patients with this CMC injury complex and the first report of open CMC dorsal dislocations and fracture-dislocations. Although early ORIF is suggested, delay of up to four weeks did not adversely affect results. PMID- 11733671 TI - Soft tissue injuries with the use of safe corridors for transfixion wire placement during external fixation of distal tibia fractures: an anatomic study. AB - OBJECTIVES: To determine which soft tissue structures are at risk and when joint violation can occur during small wire placement for hybrid external fixation of distal tibial fractures while adhering to published guidelines. DESIGN: Cadaver anatomic experiment. SETTING: University orthopaedic program. SUJBECTS: Five embalmed cadavers. INTERVENTION: Placement of small wire transfixion pins in the distal tibia. MAIN OUTCOME MEASUREMENTS: Dissection and measurements. METHODS: Four orthopaedic surgeons were shown diagrams that have been widely accepted as allowing for placement of transfixion pins in the distal tibia through safe corridors. Each of the orthopaedic surgeons was then asked to place two transfixion pins into each of five cadaver legs in a position that would provide stable external fixation of the metaphysis to the diaphysis with a circular fixator (forty pins total) for a distal tibial fracture within five centimeters of the plafond. The specimens were dissected, and pins impaling neurovascular structures, tendons, or the ankle capsule were recorded. The superior capsular synovial reflections were measured from the anterior joint line and the tip of the medial malleolus. These measurements were also performed on arthrograms of two extremities before their dissection. RESULTS: Fifty-five percent of the pins placed impaled at least one tendon that crosses the ankle joint. Neurovascular structures that were impaled included the saphenous vein (+/-10.5 percent) and the superficial peroneal nerve (+/-7.5 percent). One pin violated the superior capsular synovial reflection, which was an average of thirty-two millimeters (+/ 1.58 millimeters) from the tip of the medial malleolus and twenty-one millimeters (+/-1.63 millimeters) from the anteromedial joint line. CONCLUSIONS: This study shows that tendons and neurovascular structures above the ankle are at risk during small transfixion pin placement, even when using safe corridors. Pins placed within two centimeters of the anterior joint line or three centimeters from the medial malleolus may be intracapsular. PMID- 11733672 TI - Limited bone loss in the hip and heel after reamed intramedullary fixation and early weight-bearing of tibial fractures. AB - OBJECTIVE: To determine changes in bone mineral density at the hip and calcaneus in the injured and uninjured sides after surgery for tibial fractures. DESIGN: Prospective, longitudinal study. SETTING: University hospital. PATIENTS AND METHODS: Twenty-six patients with a tibial fracture were treated with a reamed and statically locked intramedullary nail with early weight-bearing. Dual-energy x-ray absorptiometry at the proximal femur and quantitative ultrasonography at the calcaneus were performed bilaterally after surgery and after three, six, and twelve months. RESULTS: At three months, the bone mineral density in the trochanteric region in the injured limb had decreased almost 4 percent (p < 0.007), a reduction still present at twelve months, whereas in the femoral neck, the reduction was approximately 2.5 percent at twelve months (p < 0.03). The stiffness index and speed of sound at the calcaneus on the injured side decreased (p < 0.03 and p < 0.008, respectively) during the first three months, whereas at twelve months, there was no significant difference when compared with those measured directly after surgery. There were no significant changes at any time in the proximal femur or calcaneus of the uninjured limb. CONCLUSION: Treatment with a reamed statically locked intramedullary nail and early weight-bearing resulted in limited bone loss in the proximal femur and calcaneus of the injured limb. After twelve months, the bone properties at the heel were restored, whereas there was still a small reduction at the hip. Compared with previous studies of patients with similar fractures treated with casts and restricted weight-bearing, the loss in bone mineral density was less and not as longstanding. PMID- 11733674 TI - Percutaneous skeletal joysticks for closed reduction of femoral shaft fractures during intramedullary nailing. AB - Closed locked intramedullary nailing remains the most common method for treating femoral shaft fractures. At times a closed reduction of these fractures can be difficult. A percutaneous skeletal joystick represents a simple method that can aid in the reduction maneuver. PMID- 11733673 TI - Postoperative wound infection after instrumentation of thoracic and lumbar fractures. AB - OBJECTIVE: To assess the risk of infection in trauma patients undergoing surgical intervention with instrumentation for thoracic and lumbar fractures. DATA SOURCES: A case series of 235 consecutive patients who sustained thoracic and lumbar fractures seen at Tampa General Hospital in Tampa, Florida between 1986 and 1997. STUDY SELECTION: 117 patients of the 235 consecutive patients included in the case series underwent surgical intervention; of these patients, twelve were identified as having acute postoperative wound infections. DATA EXTRACTION: Of those patients treated with operative decompression and internal fixation, the authors identified and studied those with an acute wound infection. These patients were analyzed for risk factors and infection management. DATA SYNTHESIS: Twelve (10 percent) patients with acute postoperative wound infections were identified. These included nine deep and three superficial infections. This provides an overall infection rate of 10 percent (12 of 117). Of these, there were three infections in twenty-one patients undergoing anterior spinal procedures. Only two of the twelve patients had pure cultures of gram-positive organisms (2 Staphylococcus aureus). Cultures from eight (67 percent) patients showed multiple organisms. There was a significantly (P < 0.05) higher risk of infection in the patients with a complete neurologic injury 41 percent (7/17) as compared with those with no deficit or incomplete injuries 5.0 percent (5/100). CONCLUSIONS: The overall risk of infection is higher in the trauma patient than in the elective surgery population. Those patients with a complete neurologic deficit are at a greater risk. Aggressive and early intervention can help contribute to a favorable outcome. PMID- 11733675 TI - Early pin failure following external femoral fracture fixation in a head-injured child. AB - External fixation facilitates nursing care and early rehabilitation of head injured children with femoral shaft fractures. We report an unusual complication of early fatigue failure of the external fixator pins without the patient ever bearing weight. PMID- 11733676 TI - Supracondylar distal femoral nonunions treated with a megaprosthesis in elderly patients: a report of two cases. AB - The purpose of this paper is to report the use of total knee arthroplasty, a megaprosthesis, as a treatment in elderly patients who have a persistent nonunion of a supracondylar femur fracture. This case report includes two elderly patients who sustained supracondylar femur fractures that failed to unite with standard operative fixation methods. Despite multiple procedures during a long period, patients had a persistent nonunion. Both patients underwent total arthroplasty with a cemented kinematic rotating hinge and had significant clinical improvement. The Hospital for Special Surgery (HSS) knee scores increased from fifty-four points to seventy points in one patient and forty-two points to seventy-three points after surgery in the other patient. Both patients had excellent range of motion after surgery. A cemented megaprosthesis appears to be a viable treatment option for persistent nonunions of supracondylar femur fractures in elderly patients. It is well tolerated and permits early ambulation and return to activities of daily living. PMID- 11733677 TI - One-stage treatment of open tibial fracture in a child using a distally based sural artery flap: a case report. AB - Open fractures in children have a high level of morbidity and require early treatment. This case describes the successful treatment of a child who sustained an open tibial fracture with soft tissue loss. The fracture was stabilized with a monolateral external fixator devised in our clinic, and the soft tissue loss was covered with a distally based sural artery flap. The flap is simple, can be done quickly, and a surgeon does not need microsurgical or specialty training to perform the operation. This combined use of external fixation and distally based sural artery flap is a straightforward technique in distal tibial open fractures of children with soft tissue loss. PMID- 11733678 TI - Compartment syndrome of the well leg as a result of the hemilithotomy position: a report of two cases and review of literature. AB - Compartment syndrome in the well leg as a complication of prolonged positioning in a hemilithotomy position is a serious complication that is rarely reported in the orthopaedic literature. A similar entity has been well described in urologic, gynecologic, and general surgery literature but, to the authors' knowledge, has been reported in only seven patients in the orthopaedic literature. The authors report two cases of unilateral compartment syndrome in a well leg during femoral nailing of the contralateral leg. Risk factors, theories of pathogenesis, and preventive measures are identified and discussed. PMID- 11733679 TI - Limitation of flexor tendon excursion by heterotopic ossification after isolated flexor tendon laceration. AB - A patient, one year after flexor digitorum superficialis/profundus repair in the left index finger, was diagnosed with heterotopic ossification involving the palmar surface of the proximal phalanx creating a secondary proximal interphalangeal joint contracture. A Compass PIP Hinge facilitated the treatment. Flexor tendon excursion improved, and active range of motion increased from 60 to 90 degrees before surgery to 30 to 105 degrees 20 months after surgery. Ectopic bone involvement of the hand is rare. This article reports a successful treatment for a unique complication of flexor injury and repair. PMID- 11733680 TI - Trimalleolar ankle fracture. PMID- 11733681 TI - Surgical options for the treatment of severe tibial pilon fractures. PMID- 11733682 TI - Differentiation of GABA(A) receptors in subcortical heterotopias: subunit distribution of displaced cortex reflects original commitment. AB - Recent evidence suggests that the expression of GABA(A) receptor subunits is determined by an early innate program which can be further modified by thalamic input and local factors. We analyzed the GABA(A) subunit distribution in experimentally induced subcortical heterotopia which are a subgroup of neuronal migration disorders. Heterotopias consist of clusters of neurons which have stopped migration early, before they have reached their final commitment and well before thalamic afferents have reached their targets. Immuno- histochemical analyses of five important GABA(A) receptor subunits revealed an expression pattern typical for upper cortical layers reflecting the original commitment of the heterotopic neurons. These results point towards detailed innate determinants of cell fate which even contain information on receptor subunit distribution and are not affected by ectopic positioning. PMID- 11733683 TI - Catecholamines and opioid peptides increase in plasma in humans during possession trances. AB - Naturally induced possession trances have been observed in healthy people of many societies. The neurophysiological basis of this phenomenon remains unknown, however, because of the difficulty in accessing subjects in trances due to their sacred context. In the present study, we measured the plasma levels of several neuroactive substances from subjects exhibiting or lacking possession trance characteristics during Balinese dedicatory dramas under natural conditions. The trance group exhibited significant increases in plasma concentrations of noradrenaline, dopamine and beta-endorphin, compared with controls who performed the same actions as the trance group. The present finding suggests that catecholamines and opioid peptides are involved in possession trances. PMID- 11733684 TI - Visual categorical perception by rats with temporal, striate, or sham ablations. AB - Rats were trained to discriminate a 0 degrees stripe from a 90 degrees stripe in a two choice water maze. They were prepared with either Te2/3, partial striate (PS), or sham lesions and retrained on the preoperative discrimination. In two separate experiments, excellent savings was observed for all groups. Next, trials were administered with novel stripe orientations defined as either between- or within-category problems. Performance accuracy eroded rapidly for all groups in the first experiment, and no between-group differences were observed. In the second experiment, each session with categorical stimuli was preceded by four reminder trials with the original stimuli. This improved accuracy for all groups, but it was found that animals with PS lesions, not animals with T2/3 lesions, were impaired on between-category judgements. The impairment was not secondary to a disruption of basic visual sensory processing or significantly larger lesions relative to the Te2/3 group. As is the case for monkeys, accuracy with within category stimuli was inferior to between-category stimuli for all groups. Possible reasons for this inter-species difference are discussed. PMID- 11733685 TI - Perceptual and motor mechanisms for mental rotation of human hands. AB - We measured brain potentials from human subjects performing a mental rotation task requiring right-left judgments of misoriented hands, and a control task requiring palm-back judgments of the same stimuli. High-density, 128-channel event-related potentials (ERPs) were recorded from 16 normal, right-handed subjects. There was a main effect of task at five different latencies: 148 ms (occipital), 180 ms (parietal), 388 ms (vertex), 556 ms (central-parietal), and 900 ms (vertex). Source estimations derived from topographic data indicate that frontal brain regions were strongly activated after 300 ms in the control task, but not until about 900 ms in the rotation task. We conclude that the neural computations underlying mental hand rotation may be recruited from relatively early stages of visuo-perceptual analysis; these early computations influence subsequent processing within a parietal-prefrontal system for the integration of perception with action. PMID- 11733687 TI - SOCS1 regulates interferon-gamma mediated sensory neuron survival. AB - Differentiation and survival of sensory neurons is regulated by factors such as NGF and LIF. Regulation of signal transduction pathways downstream of such factor signalling by suppressor of cytokine signalling (SOCS) proteins, which negatively regulate the JAK/STAT pathway, may modulate biological outcome. In this study, SOCS1 regulation of growth factor mediated sensory neuron survival was examined. SOCS1 expression by sensory neurons was up-regulated by IFNgamma. Survival of sensory neurons from SOCS1 null mice in NGF or LIF was similar to wildtype mice. IFNgamma partially supported survival of wildtype neurons but supported survival of SOCS1 null neurons as effectively as NGF or LIF. Thus it appears that SOCS1 is a major regulator of sensory neuron responses to the inflammatory cytokine, IFNgamma. PMID- 11733686 TI - Long-term trans-synaptic glial responses in the human thalamus after peripheral nerve injury. AB - Limb denervation leads to reorganization of the representational zones of the somatosensory cortex. Using [11C](R)-PK11195, a sensitive in vivo marker of glial cell activation, and PET, we provide first evidence that limb denervation induces a trans-synaptic increase in [11C](R)-PK11195 binding in the human thalamus but not somatosensory cortex: these brain structures appeared morphologically normal on magnetic resonance imaging (MRI). The increased thalamic signal was detectable many years after nerve injury, indicating persistent reorganization of the thalamus. This glial activation, beyond the first-order projection area of the injured neurons, may reflect continually altered afferent activity. Our findings support the view that long-term rearrangement of cortical representational maps is significantly determined within the thalamus. PMID- 11733688 TI - Involvement of capsaicin-sensitive fibers in spinal NMDA-induced glutamate release. AB - The activation of spinal NMDA receptors can evoke glutamate release through the production of nitric oxide (NO) within the spinal cord, resulting in pain relating behavior. In this study, we investigated the involvement of capsaicin sensitive primary afferents in this phenomenon using in vivo intrathecal microdialysis. Intrathecal NMDA perfusion evoked increases in the concentrations of NO metabolises and glutamate and in pain-related behavior in both neonatal capsaicin and vehicle-treated rats. Although the degrees of increase in NO metabolises in capsaicin- and vehicle-treated rats were not significantly different, capsaicin-treated rats showed significantly smaller increases in glutamate concentration and pain-related behavior than did vehicle-treated rats. Our results showed that glutamate release from capsaicin-sensitive primary afferent terminals is involved in spinal NMDA-induced pain. PMID- 11733689 TI - Dietary soy phytoestrogen effects on brain structure and aromatase in Long-Evans rats. AB - Phytoestrogens are estrogen-like (plant-derived) molecules that protect against age-related diseases (cardiovascular disease and osteoporosis), hormone-dependent (breast and prostate) cancers and selectively bind estrogen receptors. However, little is known about the influence of phytoestrogens on brain. Using diets containing either high phytoestrogen levels, derived from soy, or very low phytoestrogens we quantified phytoestrogen concentrations of daidzein, genistein and equol in brain. We found that dietary phytoestrogens: significantly decrease body and prostate weights, do not alter brain aromatase levels and significantly change during adulthood the structure of the sexually dimorphic brain region (i.e. anteroventral periventricular nucleus; AVPV) in male, but not in female rats. Since most commercial animal diets contain significant concentrations of phytoestrogens their influence on brain structure should be considered. PMID- 11733690 TI - Sex-related suppression of reflex jaw muscle activity by peripheral morphine but not GABA. AB - The present study examined the effect of peripherally applied morphine and GABA on jaw muscle electromyographic activity reflexly evoked by co-injection of glutamate into the temporomandibular joint (TMJ) of lightly anesthetized Sprague Dawley rats of both sexes. In male but not female rats, morphine significantly suppressed glutamate-evoked jaw muscle activity in a dose-dependent and naloxone reversible manner. The median suppressive dose (+/- s.e.) for male rats was 12.7 +/- 3.1 microg (digastric muscle) and 12.6 +/- 1.3 microg (masseter muscle). GABA (5 micromol) significantly reduced glutamate-evoked muscle activity in both sexes. These data suggest that female rats are considerably less sensitive than male rats to the suppressive effects of peripherally applied morphine, but both sexes are equally affected by peripherally applied GABA. PMID- 11733691 TI - Impact of signal-to-noise on functional MRI of the human amygdala. AB - The impact of signal-to-noise (SNR) on fMRI of the amygdala was investigated during a picture encoding task. The SNR value required to observe reliable activation was determined by computer simulations. Blood oxygen level-dependent (BOLD) sensitivity maps were generated to indicate brain regions with sufficient SNR to test the statistical hypotheses. The results showed that the medial aspect of the amygdala had insufficient SNR to detect a 1% peak BOLD signal change for a t-test comparison in a majority of subjects. None of these subjects showed activation in regions with unacceptable SNR values, indicating a low false positive rate. Furthermore, hemispheric asymmetries in the BOLD sensitivity maps mirrored asymmetries in the activation patterns. Impoverished SNR was also found in the basal forebrain and orbitofrontal cortex. These findings emphasize the importance of considering SNR when interpreting fMRI results in the limbic forebrain. PMID- 11733692 TI - Melatonin protects against the effects of chronic stress on sexual behaviour in male rats. AB - The effects of chronic mild stress (CMS) on both sexual behaviour and wet dog shakes (WDS), a serotonergic type 2A (5-HT2A) receptor-mediated behaviour, were explored in the male rat. In addition, the possible attenuation of these effects by chronic treatment with melatonin, a putative 5-HT2A antagonist, was examined. The CMS procedure resulted in a significant increase in WDS and an overall decrease in all aspects of sexual behaviour. Concurrent melatonin administration attenuated the CMS-induced effects on sexual behaviour, but not the effects on either spontaneous WDS or WDS in response to the 5-HT2A agonist 1-(2,5-dimethoxy 4-iodophenyl)-2-aminopropane, suggesting a mechanism of action other than exclusive 5-HT2A antagonism. These results are the first to demonstrate that melatonin significantly protects against the detrimental effects of a chronic stressor on sexual behaviour. PMID- 11733693 TI - Altered calcium dynamics in cultured cerebellar cells from IP3R1-deficient mice. AB - Intracellular calcium release in response to bath-applied quisqualate or caffeine was examined in cerebellar primary cultures of type-1-inositol-1,4,5 trisphosphate-receptor (IP3R1)- deficient mice. Under [Ca2+]o-free conditions, calcium release in response to 10 microM quisqualate was significantly reduced in Purkinje cells, but was unaffected in granule cells, suggesting that different subtypes of IP3 receptors contribute to the metabotropic glutamate response in these cells. In addition, calcium release in response to 10 mM caffeine under [Ca2+]o-free conditions was not impaired in cerebellar cells, suggesting that IICR and CICR are independently regulated in cerebellar cells. Moreover, in both wild-type and homozygous mutant mice, CICR in granule cells was approximately 6 times greater than in Purkinje cells. PMID- 11733694 TI - Histamine-induced itch converts into pain in neuropathic hyperalgesia. AB - Physiologically, itch and pain are transmitted in separate specific peripheral C units and central afferent pathways. Some neuropathic pain patients with intact but sensitized (irritable) primary C-nociceptors have spontaneous pain, heat hyperalgesia, static and dynamic mechanical hyperalgesia. The question was whether cutaneous histamine application induces pain in these patients. For comparison histamine was applied into normal skin experimentally sensitized by capsaicin. Histamine application in the capsaicin-induced primary or secondary hyperalgesic skin did not change the intensity and quality of capsaicin pain. Itch was profoundly inhibited. Conversely, histamine application in neuropathic skin induced severe increase in spontaneous burning pain but no itch. In neuropathies irritable nociceptors may express histamine receptors or induce central sensitization to histaminergic stimuli so that itch converts into pain. PMID- 11733695 TI - Parietal gamma-band activity during auditory spatial precueing of motor responses. AB - Magnetoencephalographic gamma-band activity (GBA) was used to investigate synchronization of cortical networks in putative auditory dorsal stream areas during the transformation of auditory spatial information into motor preparation. GBA was compared between lateralized vowels precueing either ipsi- or contralateral responses in two experiments with randomized versus blocked task presentation. In both studies, parietal GBA was higher for the contralateral than the ipsilateral precues. Spectral amplitudes at 54-64 Hz were maximal at 120 ms post precue onset during randomized presentations, while in the blocked task 62 72 Hz activity was present at precue onset and less peaked. These findings suggest a fast activation of parietal networks when auditory spatial precues are used to plan contralateral responses. PMID- 11733696 TI - Cortical alpha 1-adrenergic regulation of acute and sensitized morphine locomotor effects. AB - The role of alpha1-adrenergic transmission was tested on locomotor effects of acute or repeated morphine (5 mg/kg, i.p.) administration. Prazosin, an alpha1 adrenergic antagonist, administered 30 min before morphine, either systemically (0.5 mg/kg, i.p.) or locally and bilaterally into the prefrontal cortex (200 pmol/side) reduced the stimulatory influence of morphine on locomotion. The progressive increase of the locomotor response induced by repeated morphine injections was blocked by a prazosin pretreatment but not the behavioral sensitization on the test day. These data suggest that blockade of cortical alpha1-adrenergic receptors reduces the expression of acute and sensitized locomotor responses to morphine, but does not prevent the induction of behavioral sensitization. PMID- 11733697 TI - Extracellular signal-regulated kinase and c-Jun N-terminal protein kinase in ischemic tolerance. AB - The alterations and involvement of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal protein kinase (JNK) activation were examined in the hippocampal CA1 region in a rat model of global brain ischemic tolerance. Western blotting study showed that ERK activation (diphosphorylation) level was decreased (3.75-, 0.56-, and 0.23-fold vs sham control) and JNK activation level was increased (3.82-, 4.63-, and 5.30-fold vs sham control) 3 days after more severe ischemic insults (6 min, 8 min, and 10 min of ischemia, respectively). These alterations were significantly prevented by pretreatment with preconditioning ischemia, which also provided neuronal protection against ischemic injury. Inhibition of ERK activation after preconditioning ischemia by PD98059, a specific ERK kinase inhibitor, significantly prevented the inhibitory effects of preconditioning ischemia on both JNK activation and ischemic injury. The results suggest that ERK activation after preconditioning ischemia may result in the prevention of JNK activation and thus be involved in the protective responses in ischemic tolerance in hippocampal CA1 region. PMID- 11733698 TI - Stronger reactivity of the human primary motor cortex during observation of live rather than video motor acts. AB - The monkey premotor cortex contains neurons that are activated both when the monkey performs motor acts and when he observes actions made by others. A similar mirror neuron system, involving several brain areas, has been found in humans. We recorded neuromagnetic oscillatory activity from the primary motor cortex of 10 healthy subjects when they observed live and videotaped finger movements. The left and right median nerves were stimulated alternatingly and the poststimulus level of the approximately 20 Hz rhythm was quantified. Compared with the rest condition, the approximately 20 Hz rhythm was dampened 15-19% more when the subjects observed live rather than videotaped hand movements, indicating stronger activation of the primary motor cortex. These results suggest that the human mirror neuron system differentiates natural and artificially presented movements. PMID- 11733699 TI - Biphasic effects of orchidectomy on calcitonin gene-related peptide synthesis and release. AB - We hypothesize that the decline of male gonadal hormones may play a role in age related decrease of calcitonin gene-related peptide (CGRP) synthesis and release. Orchidectomized rats were raised with or without testosterone replacement and CGRP levels in serum and some tissues as well as the perfusate from the isolated mesenteric arterial bed (MAB) were measured at 1, 2 and 4 months after orchidectomy. CGRP levels of serum and tissues, and CGRP release from MAB were significantly elevated after 1 month and decreased after 4 months in orchidectomized rats. The changes were restored by testosterone replacement. Our results indicate that the age-related decline of testosterone might contribute to the age-related decrease of CGRP synthesis and release. PMID- 11733700 TI - Metabotropic purinoreceptors in rat dorsal horn neurones: predominant dendritic location. AB - Elevations of cytosolic free Ca2+ concentration ([Ca2+]i) induced by addition of ATP have been compared in rat dorsal horn neurones in slices and after their isolation. ATP application induced in neurones in situ a rise of [Ca2+]i by 201 +/- 12 nM. In Ca2+-free external solution the rise was 156 +/- 14 nM (n = 45 of 76), indicating the presence of active purinergic metabotropic receptors in about 59% of neurones. [Ca2+]i transients induced by 2MeSATP in Ca2+-free external solution were completely abolished by 10 microM PPADS, indicating that some of the corresponding receptors are of the P2Y1 type. In acutely isolated neurones which lost their dendrites, there were no metabotropic response. The results confirm the presence of metabotropic postsynaptic purinoreceptors located in the dendritic tree of dorsal horn neurones. PMID- 11733701 TI - Vasopressin differentially modulates noradrenaline release in the rat supraoptic nucleus. AB - Vasopressin is released not only from axon terminals in the neurohypophysis but also from soma/dendrite regions in the supraoptic nucleus. In order to investigate presynaptic roles of dendritically released vasopressin, we examined effects of local application of vasopressin upon noradrenaline release within the supraoptic nucleus by a microdialysis method. Noradrenaline release within the supraoptic nucleus was facilitated by local perfusion with high K+ or an NMDA receptor antagonist. Vasopressin augmented noradrenaline increase after high K+ but reduced it after an NMDA receptor antagonist, AP-5. The results suggest that dendritically released vasopressin modulates noradrenaline release within the supraoptic nucleus in a bimodal fashion. PMID- 11733702 TI - Expression of tenascin R and J1 mRNA in motoneurons after a traumatic lesion in the spinal cord. AB - We demonstrate, using in situ hybridization, that mRNA for the anti-adhesive molecules tenascin R and J1 in the adult rat spinal motoneurons are down regulated rapidly as a reaction after a ventral funiculus lesion. Tenascin-R was significantly down-regulated at day 1 and normalized after 3 weeks. Tenascin-J1 declined to its lowest value at day 3 and returned to the initial level after 3 weeks. In adjacent sections, the distribution of macrophages was studied with immuno histochemistry. The density of macrophages reached a maximum 3 days after the injury. Thus, the density of macrophages appeared to be inversely related to the level of tenascin mRNA. These data are compatible with the notion that neuronal tenascins may modulate the adhesion of perincurial inflammatory cells. PMID- 11733703 TI - Tyrosine kinase signaling involved in glutamate-induced astrocyte proliferation. AB - Proliferation of astrocytes is a common response of the CNS to injury and disease. The mechanisms controlling the proliferation of astrocytes are of great interest. In this paper, the signaling pathways underlying glutamate-induced astrocyte proliferation are investigated. Glutamate stimulates the proliferation of non-synchronized, subconfluent cultures of rat cortical astrocytes. Glutamate induced cell proliferation is not prevented by inhibitors of G protein, protein kinase A, protein kinase C, phosphatidylinositol 3 kinase, extracellular signal regulated kinase, or phospholipase A2. However, the tyrosine kinase inhibitors Genistein and Herbimycin A inhibit the glutamate-induced proliferation. Moreover, this proliferation is mediated by the activation of glutamate metabotropic receptors. These results suggest that glutamate induces astrocyte proliferation through a tyrosine kinase pathway. PMID- 11733704 TI - Neural phenotype expression of cultured human cord blood cells in vitro. AB - Neural stem cells have been proposed as useful vectors for treating diseases in the CNS, but their utility is severely limited by lack of accessibility. Brain development is ongoing extensively in early postnatal life. However, it is unclear whether stem cells that differentiate into neurons exist in the blood during early postnatal life. We showed in this experiment that neural markers (NeuN, neurofilament, MAP2, GFAP) are expressed and long cytoplasmic processes are elaborated in the cultured human cord blood monocytes prepared from newborn umbilical blood. These results suggest that stem cells in human cord blood may be potential sources of neurons in early postnatal life. We suggest that the neonatal blood system functions as a circulating pool of different types of stem cell. PMID- 11733705 TI - Sympathetic sprouting in sensory ganglia depends on the number of injured neurons. AB - We examined whether the extent of sympathetic sprouting in the dorsal root ganglion was a function of the number of injured nerve fibers. We compared two groups of rats. One group was subjected to unilateral superior and inferior caudal trunk transections at the level between the S1 and S2 spinal nerves (S-I group) and the other group was subjected to unilateral superior caudal trunk transection at the same level (S group). Immunohistochemical staining with tyrosine hydroxylase (TH) antibody of the S1 DRG revealed that the degree of TH immunoreactive fibers was more extensive in the S-I group than in the S group. However, there was no difference in the severity of neuropathic pain behaviors between the two groups. These results suggest that the extent of sympathetic sprouting in the DRG following peripheral nerve injury is proportionally related to the amount of injured nerve fibers, but not related to the degree of neuropathic pain behaviors. PMID- 11733706 TI - Variations in extracellular monoamines in the prefrontal cortex and medial hypothalamus after modafinil administration: a microdialysis study in rats. AB - The role of brain amines in mediating the effects of the wake-promoting agent modafinil, used in the treatment of sleepiness associated with narcolepsy is still uncertain. Therefore we studied the effects of modafinil on extracellular serotonin (5-HT), dopamine (DA) and noradrenaline (NA), in rat prefrontal cortex and in the medial hypothalamus area. Modafinil (128 mg/kg i.p.) significantly increased waking in the first 4 h of EEG sleep recording. This cortical and behavioral activation was associated with an initial increase in extracellular 5 HT, DA and NA during the first 60 min following modafinil administration. In the prefrontal cortex, 5-HT release remained high for 3 h after modafinil administration. In contrast, in the hypothalamus, only NA release was enhanced while DA and 5-HT levels remained low. In a first step, modafinil may generate waking partly via cortical monoamine release, particularly DA and 5-HT, and also hypothalamic NA. In a second step, maintenance of waking might depend on hypothalamic NA. PMID- 11733707 TI - Stimulation rate influences frequency tuning characteristics of inferior colliculus neurons in the little brown bat, Myotis lucifugus. AB - Previous studies of frequency selectivity have investigated unit's responses to tonal stimuli widely separated in time to minimize inter-stimulus interaction. The results of such studies are assumed to accurately portray the cell's frequency selectivity. The goal of the present study was to investigate the frequency tuning characteristics of neurons in the inferior colliculus (IC) of the little brown bat (Myotis lucifugus) to tone pulses presented at higher rates. Our results indicate that the frequency response properties of central auditory neurons at low stimulation rates do not necessarily reflect the units' frequency response properties to sounds presented at higher, more behaviorally relevant rates. Specifically, IC neurons often show greater frequency selectivity at higher stimulation rates, which presumably confers a greater perceptual frequency resolution. PMID- 11733708 TI - Functional MRI of cortical activations induced by transcranial magnetic stimulation (TMS). AB - The effects of repetitive transcranial magnetic stimulation (rTMS) on human brain activity and associated hemodynamics were investigated by blood-oxygenation-level dependent (BOLD) MRI using echo-planar imaging at 2.0 T. Apart from bilateral activation of the auditory cortex by the audible rTMS discharges (23 bursts, 1 s duration, 10 Hz, 10-20 s interstimulus intervals), BOLD responses were restricted to cortical representations of actual finger movements performed either voluntarily or evoked by suprathreshold rTMS of the motor cortex. Neither subthreshold rTMS of the motor cortex nor suprathreshold rTMS of the lateral premotor cortex induced a detectable BOLD response. These findings suggest that neuronal depolarization as induced by rTMS modulates the spiking output of a brain area but does not automatically alter cerebral blood flow and oxygenation. The observation of BOLD MRI activations probably reflects the afferent intracortical processing of real movements. PMID- 11733709 TI - Excessive sugar intake alters binding to dopamine and mu-opioid receptors in the brain. AB - Palatable food stimulates neural systems implicated in drug dependence; thus sugar might have effects like a drug of abuse. Rats were given 25% glucose solution with chow for 12 h followed by 12 h of food deprivation each day. They doubled their glucose intake in 10 days and developed a pattern of excessive intake in the first hour of daily access. After 30 days, receptor binding was compared to chow-fed controls. Dopamine D-1 receptor binding increased significantly in the accumbens core and shell. In contrast, D-2 binding decreased in the dorsal striatum. Binding to dopamine transporter increased in the midbrain. Opioid mu-1 receptor binding increased significantly in the cingulate cortex, hippocampus, locus coeruleus and accumbens shell. Thus, intermittent, excessive sugar intake sensitized D-1 and mu-1 receptors much like some drugs of abuse. PMID- 11733710 TI - External modulation of visual perception in humans. AB - Static and dynamic contrast sensitivities (sCS and dCS) were evaluated before, during, immediately after and 10 min after anodal and cathodal transcranial direct current stimulation (tDCS) applied to the occipital cortex of 15 healthy subjects. Using 4 c/d spatial and 4 Hz temporal frequencies significant sCS and dCS loss was found during and immediately after 7 min cathodal stimulation while anodal stimulation had no effect. Ten minutes after the end of the stimulation the sCS and dCS values had reached the baseline levels. Our results show that primary visual functions, such as contrast detection can be transiently altered by transcranial weak direct current stimulation, most probably modulating neural excitability, as has been shown in the motor cortex previously. The present study also support the view that this method using weak current can be a non-invasive promising tool to induce reversible focal changes in the nervous system. PMID- 11733711 TI - P-glycoprotein and multidrug resistance-associated protein are involved in the regulation of extracellular levels of the major antiepileptic drug carbamazepine in the brain. AB - Despite considerable advances in the pharmacotherapy of epilepsy, about 30% of epileptic patients are refractory to antiepileptic drugs (AEDs). In most cases, a patient who is resistant to one major AED is also refractory to other AEDs, although these drugs act by different mechanisms. The mechanisms that lead to drug resistance in epilepsy are not known. Recently, over-expression of multidrug transporters, such as P-glycoprotein (PGP) and multidrug resistance-associated protein (MRP), has been reported in surgically resected epileptogenic human brain tissue and suggested to contribute to the drug resistance of epilepsy. However, it is not known to what extent multidrug transporters such as PGP or MRP are involved in transport of AEDs. In the present study, we used in vivo microdialysis in rats to study whether the concentration of carbamazepine in the extracellular fluid of the cerebral cortex can be enhanced by inhibition of PGP or MRP, using the PGP inhibitor verapamil and the MRP inhibitor probenecid. Local perfusion with verapamil or probenecid via the microdialysis probe increased the extracellular concentration of carbamazepine. The data indicate that both PGP and MRP participate in the regulation of extracellular brain concentrations of the major AED carbamazepine. PMID- 11733713 TI - Activation of Fas and caspase 3 precedes PrP accumulation in 87V scrapie. AB - The sequence of events involved in the neurodegeneration caused by transmissible spongiform encephalopathies is not yet known. Using a murine scrapie model in which neurodegeneration in the hippocampus is restricted to the CA2, we show an up-regulation of the proapoptotic markers Fas and caspase 3 early in the incubation period prior to disease-specific prion protein (PrP) deposition and clinical signs. These results suggest that activation of Fas and caspase 3 are involved in the early pathological sequence of events during murine scrapie, and that these proapoptotic markers may be a specific method for early detection of neurodegeneration. PMID- 11733712 TI - Levels of word processing and incidental memory: dissociable mechanisms in the temporal lobe. AB - Word recall is facilitated when deep (e.g. semantic) processing is applied during encoding. This fact raises the question of the existence of specific brain mechanisms supporting different levels of information processing that can modulate incidental memory performance. In this study we obtained spatiotemporal brain activation profiles, using magnetic source imaging, from 10 adult volunteers as they performed a shallow (phonological) processing task and a deep (semantic) processing task. When phonological analysis of the word stimuli into their constituent phonemes was required, activation was largely restricted to the posterior portion of the left superior temporal gyrus (area 22). Conversely, when access to lexical/semantic representations was required, activation was found predominantly in the left middle temporal gyrus and medial temporal cortex. The differential engagement of each mechanism during word encoding was associated with dramatic changes in subsequent incidental memory performance. PMID- 11733714 TI - Delta 9-tetrahydrocannabinol-induced alterations in limbic system glucose use in the rat. AB - The effects of Delta9-tetrahydrocannabinol (THC) on neural activity in the rat were investigated using [14C]2-deoxyglucose autoradiography to measure local cerebral glucose use. Overall, THC reduced glucose use at 5 mg/kg and slightly increased it at 1 mg/kg. Significant reductions were seen at the higher dose in regions of the hippocampus and limbic system, and in structures associated with sensory and sensorimotor process ing, consistent with the effects of THC on memory, sensory perception and motor control. Glucose use was increased in the shell of the nucleus accumbens at 1 mg/kg but not reduced at 5 mg/kg. This may reflect increases in activity in mesolimbic reward pathways which at the higher dose are mitigated by reduced glutamatergic outflow from the hippocampus. PMID- 11733715 TI - Repolarization of the plasma membrane shapes NMDA-induced cytosolic [Ca2+ ] transients. AB - After inactivation of NMDA receptors, restoration of basal cytosolic [Ca2+] ([Ca2+]c) is delayed. This may be caused by Ca2+ influx via reverse Na/Ca exchange or voltage-gated Ca2+ channels, and/or by Ca2+ efflux from internal stores. Monitoring of [Na+]c, [Ca2+]c, and plasma membrane potential in cultured cerebellar granule cells showed that repolarization of the plasma membrane and inactivation of voltage-gated Ca channels plays the most critical role in restoration of low [Ca2+]c following NMDA receptor inactivation. During NMDA receptor activation, however, an Na-dependent mechanism enhanced NMDA-induced elevation in [Ca2+]c. This mechanism did not involve Na,K-ATPase activation by Na+, because it operated even when Na,K-ATPase was inhibited. PMID- 11733716 TI - Cognitive demands of executing postural reactions: does aging impede attention switching? AB - A new dual-task paradigm was used to investigate age-related differences in attentional dynamics during rapid balancing reactions evoked by small, unpredictable antero-posterior platform movements. The perturbations were delivered while subjects performed a continuous visuo-motor pursuit-tracking task. Onset of significant deviation in tracking was inferred to indicate switching of attentional resources between tracking and balancing tasks. Although tracking deviation was equally likely to occur subsequent to postural perturbation in healthy young and older adults, deviation onset was delayed, on average, by 67% (123 ms) in the older subjects. Delay in onset of tracking deviation correlated with subsequent delay in generating the peak stabilizing postural response at the ankle. These results suggest that impaired attentional dynamics may exacerbate postural instability in older adults. PMID- 11733717 TI - Hyperbaric oxygen treatment decreases post-ischemic neurotrophin-3 mRNA down regulation in the rat hippocampus. AB - The therapeutic effect of hyperbaric oxygen (HBO) on ischemic injury was investigated using in situ hybridization to detect the mRNA expression of neurotrophin-3 (NT-3), which is thought to play a crucial role in protecting against neuronal death induced by brain ischemia. The rats under investigation were subjected to 10 min transient forebrain ischemia, and subsequently exposed to HBO (100% oxygen, 2.5 atm absolute) for 2 h. Levels of NT-3 mRNA in the CA1, CA2 and CA3 regions, and the dentate gyrus of the hippocampus were measured after various reperfusion periods. Neuronal death in the hippocampal CA1 region was also measured by Nissl staining, seven days post ischemia. The results demonstrated that HBO treatment significantly reduced the ischemia-induced down regulation of the NT-3 mRNA level at 4 h post ischemia, and significantly increased cell survival 7 days after reperfusion. The findings suggest that an HBO treatment maintaining the NT-3 mRNA level in the hippocampus can be beneficial to the ischemic brain within a certain time frame. PMID- 11733718 TI - Chronic exposure of kainate and NBQX changes AMPA toxicity in hippocampal slice cultures. AB - Receptor agonist/antagonist mediated modulation of the excitotoxic effect of AMPA was studied in organotypic hippocampal slice cultures. Treatment of developing cultures for 2 weeks with a subtoxic dose of 2 microM kainate reduced the toxicity of 3 microM AMPA, applied for 48 h with 24 h of recovery, as measured by cellular uptake of the fluorescent dye propidium iodide. In contrast long-term treatment with 0.3 microM of the AMPA/KA antagonist NBQX increased the susceptibility of the cultures to an even lower dose of 2 microM AMPA. The modulatory effects of long-term application of low doses of kainate and NBQX, have implications for the development and use of related drugs that aim to protect against glutamate receptor-mediated disturbances. PMID- 11733719 TI - NGF induces neurite outgrowth via a decrease in phosphorylation of myosin light chain in PC12 cells. AB - The relationship between phosphorylation of myosin light chain (MLC) and neurite outgrowth induced by nerve growth factor (NGF) was studied in PC12 cells. Inhibitors of Rho kinase, HA-1077 or Y-27632 also induced neurite outgrowth. As already reported botulinum exoenzyme C3 which inactivates Rho protein also induced neurite outgrowth. Calyeulin A, an inhibitor of phosphatase counteracted both NGF- and C3- induced neurite outgrowth. Treatments of both NGF and C3 resulted in significant and transient decrease in phosphorylated MLC. These results suggest that NGF induces neurite outgrowth of PC12 by a transient decrease in phosphorylated MLC which is brought about by activation of MLC phosphatase via inhibition of Rho-Rho kinase pathway. PMID- 11733720 TI - Psychophysical evidence for a general temporal processing deficit in children with dyslexia. AB - The hypothesis of a general (i.e. cross-modal) temporal processing deficit in dyslexia was tested by examining rapid processing in both the auditory and the visual system in the same children with dyslexia. Participants were 10- to 12 year-old dyslexic readers and age-matched normal reading controls. Psychophysical thresholds were estimated for auditory gap and visual double flash detection, using a two-interval, two-alternative forced-choice paradigm. Significant group differences were found for the auditory and the visual test. Furthermore, temporal processing measures were significantly related to word and pseudo-word reading skills. As 70% of the dyslexic readers had significantly higher thresholds than controls for both auditory and visual temporal processing, the evidence tends to support the hypothesis of a general temporal processing deficit in children with dyslexia. PMID- 11733721 TI - Delayed treatment with polynitroxyl albumin reduces infarct size after stroke in rats. AB - Nitroxides are antioxidants that are known to protect cells from oxidative damage. Polynitroxyl albumin (PNA) is a compound of human serum albumin covalently labeled with nitroxides that exhibits a prolonged half-life and an enhanced antioxidant activity. Adult male Sprague-Dawley rats were subjected to 90 min intraluminal middle cerebral artery occlusion and the drug was administered intravenously immediately or 2 h after reperfusion. The effects of the drug were evaluated 24 h after reperfusion. Infarct volume was significantly reduced in immediate (79% reduction) and delayed (53% reduction) PNA-treated groups. The efficacy of a single, delayed i.v. injection of PNA suggests that PNA has great promise in the treatment of acute human stroke. PMID- 11733722 TI - Task-switching and memory retrieval processing: electrophysiological evidence. AB - Participants attempted to retrieve phonological or imagery-based information under conditions where either the two kinds of retrieval were required in separate blocks, or where frequent switches between retrieval tasks were required within blocks. Electrophysiological indices of processes engaged in pursuit of accurate memory judgments, elicited by contrasting ERPs evoked by correctly identified new test items, differed according to retrieval task only when the tasks were completed in separate blocks. The principal conclusion suggested by these findings is that the requirement to alternate frequently between the two retrieval tasks attenuated the engagement of task-specific processes that form part of a retrieval attempt. PMID- 11733723 TI - Neuroimaging correlates of negative priming. AB - Many theoretical accounts of selective attention and memory retrieval include reference to active inhibitory processes, such as those argued to underlie the negative priming effect. fMRI was used in order to investigate the areas of cortical activation associated with Stroop interference, Stroop facilitation and Stroop negative priming tasks. The most significant activation within the negative priming task was within the inferior parietal lobule, left temporal lobe and frontal lobes. Areas of cortical activation are discussed with reference to theoretical accounts of the negative priming effect. PMID- 11733724 TI - Making sense during conversation: an fMRI study. AB - Although language is thought of as a left hemisphere function, there is increasing evidence that the right hemisphere contributes to language processing by identifying the theme of spoken and written language. Using fMRI, we examined the role played by the right and left hemispheres in making sense of a conversation. When this process involves implicit appraisal of changes in the conversation's topic, the neural network has a right hemisphere bias and includes Broca's and Wernicke's areas, their right hemisphere homologues, right dorsolateral prefrontal cortex, and the cerebellum. When making sense of conversation involves appraisal of the conversation's reasoning, however, the network includes Broca's and Wernicke's areas. Thus, right and left hemisphere systems contribute uniquely to the linguistic skills involved in making sense of a conversation. PMID- 11733725 TI - Visual spatial localization conflict: an fMRI study. AB - fMRI and ERP studies have shown that tasks comprising conflicting stimulus response associations activate a variety of cortical regions. It remains unclear whether any of these areas are activated by all conflict tasks, or whether conflict resolution is a common property of a number of distinct anatomical regions. Several regions in frontal and parietal cortex are activated by both exogenous (position) and endogenous (arrow direction) localization cues. The present event-related fMRI study used a version of the Simon task with independent positional and directional cues. The results indicated that spatial localization conflict activated pre-motor and superior parietal regions in the right hemisphere known to be involved in spatial localization, but anterior cingulate activation did not reach threshold. This suggests that conflict within a single functional modality may be processed in the region embodying that modality, and anterior cingulate may be called on only to resolve conflict between modalities. PMID- 11733726 TI - In vivo damage of CNS myelin and axons induced by peroxynitrite. AB - In multiple sclerosis (MS) the mechanisms of injury caused by peroxynitrite remain uncertain. To study histological, ultra structural and molecular alterations caused by peroxynitrite in brain, the peroxynitrite donor 3 morpholinosydnonimine was injected in rat corpus callosum. Peroxynitrite induces strong primary axonal damage with characteristics of primary acute axonopathy, together with severe myelin alteration, myelin vacuolation and demyelination, and nitrotyrosine formation as confirmed by detection of nitrosated target proteins. Administration of the peroxynitrite scavenger uric acid inhibited these effects. In vivo, peroxynitrite leads to a disorganisation of myelin and to axonal damage presenting some similarities to the formation of MS lesions. Understanding the action of peroxynitrite in this process will open new therapeutic strategies by specific inhibition of peroxynitrite formation and action. PMID- 11733728 TI - Ionotropic (P2X) receptor dynamics at single autonomic varicosities. PMID- 11733729 TI - Outcome of percutaneous coronary angioplasty (PTCA) performed in a low-volume institution by low-volume operators, evaluated by means of the one-month major adverse cardiac event rate. AB - BACKGROUND: Since an inverse relationship between percutaneous coronary angioplasty (PTCA) case-load and in-hospital major adverse cardiac events (MACE) exists, we intended to evaluate the performance of low-volume PTCA operators, during the first year of our interventional program, by applying the more accurate index represented by the MACE rate within the first month. METHODS: The data relative to both the PTCA procedure and the control visit 3-4 weeks later, were retrospectively reviewed. Death, myocardial infarction and need for revascularization were the end-points evaluated, both globally and with respect to the individual operators. RESULTS: During 1999, 61 consecutive patients (53M, 8F; mean age: 59.9+/-10.4 years) were treated by two full-trained operators. Stable angina was the indication in 75% of cases. Comorbidities as diabetes and prior revascularization, were present in 16 and 5% of cases, respectively. Multivessel procedures were performed in 33% of cases, with a total number of lesions of 84 (77% A/B1 type). Stents were implanted in 70% of cases, as a bail out in 12%. Procedural success rate was 93%. Overall one-month MACE rate was 3.3%, accounted for by 1 in-hospital emergency coronary surgery occurred to operator 1 (3.6% one-month MACE rate) and 1 elective coronary operation performed in a stable patient previously treated by operator 2 (3% one-month MACE rate). CONCLUSIONS: PTCA performed in a low-volume center by low-volume operators is not necessarily associated with a poor outcome, provided that adequate selection of low-risk cases is accomplished. Although only 52% of the Italian centers met in 1999 the recommended volume standards, reaching optimal case-load should anyway be pursued. Some time should however be conceded, provided that close monitoring of one-month MACE rate shows adequate performance of both the institution and the operators. PMID- 11733730 TI - [Evaluation of resistance to activated protein C in myocardial infarction patients]. AB - BACKGROUND: It is known that resistance to activated protein C (APCR), often associated with a single point mutation (Arg506-->Gln) in the coagulation factor V gene (factor V Leiden), is the most frequent inherited abnormality of blood coagulation. It plays a key role as pathogenetic factor of venous thromboembolism, but its association with an increased risk of arterial thrombosis is uncertain. Aim of the present study was to evaluate the prevalence of APCR in men, who suffered myocardial infarction more than 6 months earlier and without cardiovascular risk factors (hypercholesterolemia, smoking, diabetes, obesity and hypertension). METHODS: The study was carried on 20 men aged <65 years who have suffered myocardial infarction. Twenty healthy subjetcs matched for sex and age were recruited as controls. We determined: PTT, PTT-PCA (Activated Protein C), PTT-PCA/PTT, AntiThrombin III, Protein C, Protein S, Fibrinogen, Glucose, Triglycerides, Cholesterol, HDL-C, LDL-C, APO A1, APO B100, Lp(a), oxizided LDL antibody and some components of the complement system (C3c, C4). RESULTS: In the group of patients there were six subjects (30%) with APCR, while there were no subjects in the control group (0%) with APCR. Patients were subdivided into two groups: with and without APCR. Patients with APCR displayed significantly higher levels of fibrinogen (367.5+/-48.4 vs 268.3+/-37.7), LDL-C (147.0 +/-20.7 vs 125.8+/-25,6), APO B100 (133.8+/-29.8 vs 121.8+/-31.5), oxizide LDL antibody (596.8+/-357 vs 315.3+/-179.6) and C4 (45.6+/-10 vs 37.85+/-11,3). Significant positive correlations (p<0.05) were observed between PTT-PCA/PTT ratio and fibrinogen (r=0.68) and between PTT-PCA/PTT ratio and oxidized LDL antibody (r=0.61). CONCLUSIONS: In conclusion, the high prevalence of APCR (30%) in our patients seems to be important in order to carry out a primary prevention of arterial thrombosis and a secondary prevention of new thromboembolic complications. PMID- 11733731 TI - Patent ductus arteriosus closure using the new Amplatzer Duct Occluder. Preliminary results and review of the literature. AB - BACKGROUND: Patent ductus arteriosus (PDA) is the second most common congenital heart disease. A large number of surgical and transcatheter techniques for the interruption or closure of PDA has been reported. The aim of this study was to assess the immediate and short-term results of transcatheter closure of PDA using the new, self-expandable, self-centering, and repositionable Amplatzer Duct Occluder device. METHODS: We attempted occlusion of PDA with the Amplatzer Duct Occluder in seven consecutive patients, one child and six adults, four females and three males, between September 1999 and January 30th 2000. All PDAs but one were approached from the femoral venous site; the Amplatzer Duct Occluder size was selected in order to be 2 mm larger than the duct's diameter at its narrowest site and the mean PDA diameter was 5.4+/-2.5 mm (range 3-9). All patients underwent physical examination, chest X-ray and echocardiography within 48 hours and on first and third month after PDA occlusion. RESULTS: Four patients had a megaphone type (type A), and three had an elongated, conical type (type E) PDA. Four patients had immediate, complete angiographic closure of the ductus 10 minutes after the procedure, one had a trace shunt and two had small shunts which all disappeared within 48 hours. The average fluoroscopy time and procedural time were 34.4+/-10.6 min (range 21-50) and 105+/-38.9 min (range 75-190) respectively. There were no complications at follow-up. CONCLUSIONS: Transcatheter closure of PDA using the new Amplatzer Duct Occluder is an easy and effective technique. Moreover it is safe even in the presence of wide PDAs. PMID- 11733732 TI - Restenosis after carotid surgery. AB - On the basis of personal experience and reports in the literature, the paper examines the incidence of restenosis after carotid surgery, its potential morbidity and the results of its treatment. The incidence of stenosis after carotid surgery is described in a series of 253 patients given carotid TEAs in 1992-98 and followed up at 1, 3, 6 and 12 months with yearly check-ups thereafter. The group's age range was 64-70 and it included 35% females, 65% males. The initial surgical procedure was a standard TEA with or without patching. Identified by colour Doppler scan, cases of restenoses were examined angiographically in the presence of significant symptoms and haemodynamic disorders. This group constituted 7.5% of the personal series and was classified on the basis of clinical, morphological, topographical and haemodynamic criteria. Only 21% of the restenosis cases required repeat surgery: 2 classic procedures and 2 endovascular operations. No complications ensued and all these patients were symptom-free at 84 months' follow-up. A comparison of these data with reports in the literature confirms an increasing incidence of restenosis after carotid surgery in recent years, which partly reflects more accurate diagnosis. It is concluded that indications to surgery should take greater account of the lesion's histological and clinical characteristics and the increased risk of peripheral nerve damage in repeat surgery. The alternative endovascular approach will need to be examined on the basis of bigger case series and longer, more meaningful follow-ups before any definitive conclusions can be reached though the endovascular approach does seem to represent the future of treatment in this particular sector. PMID- 11733733 TI - [Indication for short stripping of the great saphenous vein. Results and indications]. AB - BACKGROUND: The aim of surgical therapy of varicose veins is the elimination of reflux from the deep to superficial system at the saphenous crosse and perforant vessel and conservation of the superficial venous system due to possible surgical procedures for arterial revascularization. This latter condition leads to an extension of indications for short stripping procedures, although the venous distal segment may undergo hypoplastic degeneration not compatible for revascularization purposes. Another important reason is the minor incidence of neurologic complication due to saphenous nerve lesion which may occur during long saphenous stripping. METHODS: From January 1994 to June 1999, we considered 233 patients (182 women, 51 men); 180 cases underwent long saphenous stripping procedures, whereas 53 a short stripping of GSV. The incidence of neurologic complications of the saphenous nerve were recorded in 11.6% of the patients treated with the standard procedure, whereas no such complication was observed in all cases treated with the short stripping procedure. RESULTS: ECD follow-up performed for a period of three months from the surgical procedure revealed the patency of the residual saphenous vein, with a minimum diameter of 3 mm, in 28 patients (56.6%). CONCLUSIONS: Our opinion is to extend the indication for short stripping of the saphenous vein to all cases where the distal saphenous trunk is not involved, when the ECD shows a pathological ostial reflux, a truncular reflux limited to the thigh, which may be associated with incontinence of the perforant vein of Dodd. PMID- 11733734 TI - Gene-mediated inhibition of the b-adrenergic receptor kinase: a new therapeutic strategy for heart failure. AB - Molecular changes that take place during the evolution of heart failure (HF), especially the well characterized beta-adrenergic receptor (betaAR) signaling abnormalities, represent attractive targets for myocardial gene therapy. The beta adrenergic receptor kinase (betaARK1 or GRK2) is a cytosolic enzyme that phosphorylates only agonist-occupied betaARs as well as other G protein-coupled receptors (GPCRs), leading to desensitization and functional uncoupling. betaARK1 levels and activity are elevated in the failing heart and therefore, it has recently been evaluated as a potential target for novel HF treatment. This review summarizes recent results obtained in transgenic mouse models as well as in animals where a betaARK1 inhibitor peptide (betaARKct) was delivered via the coronary arteries by exogenous gene transfer. These results strongly suggest that betaARK1 inhibition may represent a significant improvement in HF therapy. PMID- 11733735 TI - Provisional versus routine coronary stenting. A review. AB - Coronary stenting is the most commonly used coronary revascularization procedure due to the ease of use, safety and evidence that it has predictably better results in several coronary lesion subsets than balloon angioplasty alone. As opposed to routinely stenting all the stentable coronary lesions, in the provisional stenting approach, all the lesions would first be optimally angioplastied and a stent used only in the patients who would not have as good an outcome with balloon angioplasty alone. There are several theoretical advantages to this approach. The two different approaches are compared in this review. Retrospective studies and studies using immediate vessel recoil after optimal balloon angioplasty seemed to suggest that stent-like results after balloon angioplasty had similar target revascularization rates as stenting. However, a prospective randomized study primarily using angiography to guide provisional stenting suggests better outcome in the routine stenting strategy. If provisional stenting were to have similar long-term results as routine stenting, it has to be guided by techniques other than coronary angiography like coronary flow reserve or intravascular ultrasound. Even then, over half of the balloon angioplasty group will need stenting. Besides, based on prospective randomized studies, this strategy is not economically more attractive than the routine stenting strategy. Therefore routine stenting strategy is justifiable for most operators who use coronary angiography to guide their interventions. With further advancement in the stent technology like drug coating routine stenting may be even further attractive. PMID- 11733736 TI - Patent foramen ovale: pathophysiology and therapeutic options in symptomatic patients. AB - The foramen ovale, a remnant from the fetal circulation, remains patent through adulthood in approximately 1/4th of the general population, thus representing the most common persistent abnormality of fetal origin. In these individuals, the patent foramen ovale (PFO) permits interatrial right-to-left shunting during those periods of time when right atrial exceeds left atrial pressure. Recently, the pathophysiological aspects of the PFO have been increasingly appreciated, giving rise to disease manifestations such as paradoxical embolism, refractory hypoxemia in patients with right ventricular infarction or severe pulmonary disease, orthostatic desaturation in the setting of the rare platypnea orthodeoxia syndrome, neurological decompression illness in divers, and migraine headache with aura. Despite the growing recognition of the PFO, particularly when associated with an atrial septal aneurysm, as risk factor for paradoxical embolism, the optimal treatment strategy for symptomatic patients remains undefined. Most patients with presumed paradoxical embolism are currently treated medically with antithrombotic medications, with a paucity of data concerning the efficacy of oral anticoagulant as opposed to antiplatelet therapy. Surgical PFO closure has proved feasible, but the procedure is associated with the well known complications of cardiac surgery, and the results have been mixed with respect to stroke prevention. The recent introduction of interatrial septal occlusion devices set the stage for a minimally invasive, percutaneous approach. The present article discusses the pathophysiology of the PFO, and the advantages and drawbacks of the different therapeutic options available for symptomatic patients. PMID- 11733737 TI - [Inhibitors of the platelet's receptor GPIIb/IIIA. Progress in the treatment of acute myocardial ischemia and future trends]. AB - The possible mechanisms inducing acute coronary obstruction starting from the atherosclerotic plaque are illustrated. The effects of the inhibitors of the platelet's receptor GPIIb/IIIa and the results of the main trials performed using these inhibitors are reported. Finally, the results obtained immediately before their employment in the coronary angioplasty with or without stent's implantation are also reported. The first applications in acute ischemic stroke appear to be favourable too. PMID- 11733738 TI - [Leukocytes-endothelial interaction in vascular pathology: from host defence mechanism to promoter of vascular injury]. AB - Leukocytes play a key role in the defense of human organism from bacterial and fungal infections, taking part in the host defense mechanism against viral and neoplastic diseases. The purpose of this article is to underline the interaction between leukocytes and endothelial cells in atherosclerosis vascular damage. Actually, new evidences suggest a primary role for leukocytes, adhesion molecules, citokines and proteases to promote vascular endothelial cell injury by inflammatory process, the first step in atheromatic plaque formation. PMID- 11733739 TI - [Proarrhythmia caused by amiodarone]. AB - A woman with mitral prosthesis, treated with amiodarone i.v. (500 mg in two boluses) for an atrial tachycardia, shows repeated episodes of torsade de pointes. Although there is a connection between the intensity of electro physiological alterations due to the anti-arrhythmic therapy and pro-arrhythmic effects, there are cases in which the torsade de pointes seems to be associated with modest alterations of QT intervals or even without high plasma concentrations of drugs. In these cases a genetic predisposition for drug's metabolism alteration or greater sensibility of target organs is suggested. PMID- 11733740 TI - [Thrombophilia and thromboembolic disease. A case report]. AB - Congenital thrombophilia, specifically thrombophilia due to Leiden modified factor V as the cause of thromboembolic disease, is discussed. Thromboembolic disease is potentially fatal and disabling. A clinical case is presented of a girl of 15 with Leiden factor V thrombophilia in which all the typical thromboembolic events occurred: superficial thrombophlebitis, rapidly developing deep venous thrombosis of an extremity and ischaemic arterial thrombosis of the distal arterial trunk of the same extremity. Systemic thrombolytic therapy done quickly cleared up the acute phase of the thromboembolic event without it being necessary to resort to thrombectomy and prevented post-thrombotic sequelae involving the extremity. PMID- 11733741 TI - [Thrombolytic therapy during cardiopulmonary resuscitation for acute massive pulmonary embolism. A case report]. AB - Massive pulmonary embolism associated with cardiac arrest has an extremely high mortality in spite of cardiopulmonary resuscitation maneuvers. An early diagnosis of pulmonary embolism as cause of cardiac arrest and a rapid specific therapy able to obtain a restoration of pulmonary flow can improve the prognosis. The authors report a case of cardiac arrest for massive pulmonary embolism promptly diagnosed by echocardiography and treated by thrombolytic therapy with an initial favourable outcome. PMID- 11733742 TI - [Use of the modified Fogarty in acute occlusions of inverted venous graft. Technique proposal]. AB - The acute thrombosis of inverted venous graft, although less frequent than of the alloplastic prosthesis graft, represents a problem for the vascular surgeon that on one hand wants to restore the flow and on the other to preserve, as much as possible, the integrity of the venous endothelium without damaging the valvular apparatus. The two objectives are not possible, using a traditional Fogarty balloon catheter: the introduction from the proximal anastomosis, the only possible way for the presence of the valves, requires that, for the removal of the thrombotic material, the instrument is drawn back in a contrary way with unavoidable damage of the valves. Such disadvantage is eliminated using a modified Fogarty catheter, that allows to introduce the instrument in cranio caudal direction and to draw it back in the same way, with impossibility to stop into the bottom of the valvular border and with a minimal trauma of parietal and valvular endothelium. On the other hand, distal introduction of the traditional Fogarty catheter is difficult, if not impossible, due to the presence of the valves. The use of the Fogarty catheter from the top to the bottom of the graft is feasible after appropriate modifications of the traditional catheter that allow its introduction from the tail and to draw it back towards the periphery (with inflated balloon) according to the direction of the flow. Such modifications of the Fogarty catheter are easily feasible even on the operating table and they don't require particular devices. The technique is simple, does not require additional costs (this particular modified catheter can be, like the traditional, reusable) and allows the graft patency if the thrombosis cause is eliminated. PMID- 11733743 TI - Pediatric steroid-resistant nephrotic syndrome. PMID- 11733745 TI - Non-coding RNA genes and the modern RNA world. AB - Non-coding RNA (ncRNA) genes produce functional RNA molecules rather than encoding proteins. However, almost all means of gene identification assume that genes encode proteins, so even in the era of complete genome sequences, ncRNA genes have been effectively invisible. Recently, several different systematic screens have identified a surprisingly large number of new ncRNA genes. Non coding RNAs seem to be particularly abundant in roles that require highly specific nucleic acid recognition without complex catalysis, such as in directing post-transcriptional regulation of gene expression or in guiding RNA modifications. PMID- 11733746 TI - Accessing genetic variation: genotyping single nucleotide polymorphisms. AB - Understanding the relationship between genetic variation and biological function on a genomic scale is expected to provide fundamental new insights into the biology, evolution and pathophysiology of humans and other species. The hope that single nucleotide polymorphisms (SNPs) will allow genes that underlie complex disease to be identified, together with progress in identifying large sets of SNPs, are the driving forces behind intense efforts to establish the technology for large-scale analysis of SNPs. New genotyping methods that are high throughput, accurate and cheap are urgently needed for gaining full access to the abundant genetic variation of organisms. PMID- 11733747 TI - Genetics of autism: complex aetiology for a heterogeneous disorder. AB - Since autism was first recognized as a disorder in 1943, speculation about its aetiology has ranged from biological to psychological and back again. After twin studies during the 1970s and 1980s yielded unequivocal evidence for a genetic component, aetiological research in autism began to focus primarily on uncovering the genetic mechanisms involved. The identification of chromosomal abnormalities and Mendelian syndromes among individuals with autism, in conjunction with data from genome screens and candidate-gene studies, has helped to refine the view of the complex genetics that underlies autism spectrum conditions. PMID- 11733748 TI - The art and design of genetic screens: zebrafish. AB - Inventive genetic screens in zebrafish are revealing new genetic pathways that control vertebrate development, disease and behaviour. By exploiting the versatility of zebrafish, biological processes that had been previously obscured can be visualized and many of the responsible genes can be isolated. Coupled with gene knockdown and overexpression technologies, and small-molecule-induced phenotypes, genetic screens in zebrafish provide a powerful system by which to dissect vertebrate gene function and gene networks. PMID- 11733749 TI - Genetics of susceptibility to human infectious disease. AB - Before Robert Koch's work in the late nineteenth century, diseases such as tuberculosis and leprosy were widely believed to be inherited disorders. Heritability of susceptibility to several infectious diseases has been confirmed by studies in the twentieth century. Infectious diseases, old and new, continue to be an important cause of mortality worldwide. A greater understanding of disease processes is needed if more effective therapies and more useful vaccines are to be produced. As part of this effort, developments in genetics have allowed a more systematic study of the impact that the human genome and infectious disease have on each other. PMID- 11733750 TI - Genome sequencing networks. AB - Genome sequencing projects have been undertaken in one of three ways: in a purpose-built and professionally staffed genome centre, by a small number of traditional research laboratories or by an extensive network of traditional research laboratories that are linked by the Internet. Sequencing networks are an attractive option in many circumstances as they are easy to create, bring together diverse types of expertise, integrate the eventual users of a genome sequence with its determination and generally foster a collaborative spirit. PMID- 11733751 TI - Improving plant breeding with exotic genetic libraries. AB - Naturally occurring variation among wild relatives of cultivated crops is an under-exploited resource in plant breeding. Here, I argue that exotic libraries, which consist of marker-defined genomic regions taken from wild species and introgressed onto the background of elite crop lines, provide plant breeders with an important opportunity to improve the agricultural performance of modern crop varieties. These libraries can also act as reagents for the discovery and characterization of genes that underlie traits of agricultural value. PMID- 11733752 TI - A small fortune. PMID- 11733754 TI - Neighbourhood watch. PMID- 11733755 TI - A life or death situation? PMID- 11733758 TI - Damage control. PMID- 11733759 TI - Jelly belly. PMID- 11733761 TI - Role model. PMID- 11733762 TI - A guiding star. PMID- 11733763 TI - Switching on S phase. PMID- 11733764 TI - Family ties. PMID- 11733765 TI - Boxing day. PMID- 11733766 TI - Staying alive. PMID- 11733767 TI - CHK2 kinase--a busy messenger. AB - Checkpoint kinase 2 (Chk2) is emerging as a key mediator of diverse cellular responses to genotoxic stress, guarding the integrity of the genome throughout eukaryotic evolution. Recent studies show the fundamental role of Chk2 in the network of genome-surveillance pathways that coordinate cell-cycle progression with DNA repair and cell survival or death. Defects in Chk2 contribute to the development of both hereditary and sporadic human cancers, and earmark this kinase as a candidate tumour suppressor and an attractive target for drug discovery. PMID- 11733768 TI - Rho-family GTPases in cadherin-mediated cell-cell adhesion. AB - Cell-cell adhesions are rearranged dynamically during tissue development and tumour metastasis. Recently, Rho-family GTPases, including RhoA, Rac1 and Cdc42, have emerged as key regulators of cadherin-mediated cell-cell adhesion. Following the identification and characterization of regulators and effectors of Rho GTPases, signal transduction pathways from cadherin to Rho GTPases and, in turn, from Rho GTPases to cadherin, are beginning to be clarified. PMID- 11733769 TI - Kinetic modelling approaches to in vivo imaging. AB - The ability to visualize protein dynamics and biological processes by in vivo microscopy is revolutionizing many areas of biology. These methods generate large, kinetically complex data sets, which often cannot be intuitively interpreted. The combination of dynamic imaging and computational modelling is emerging as a powerful tool for the quantitation of biophysical properties of molecules and processes. The new discipline of computational cell biology will be essential in uncovering the pathways, mechanisms and controls of biological processes and systems as they occur in vivo. PMID- 11733770 TI - Network dynamics and cell physiology. AB - Complex assemblies of interacting proteins carry out most of the interesting jobs in a cell, such as metabolism, DNA synthesis, movement and information processing. These physiological properties play out as a subtle molecular dance, choreographed by underlying regulatory networks. To understand this dance, a new breed of theoretical molecular biologists reproduces these networks in computers and in the mathematical language of dynamical systems. PMID- 11733771 TI - The role of Fas ligand in immune privilege. AB - Immune privilege is a property of some sites in the body, whereby immune responses are limited or prevented. One explanation that has been proposed for this phenomenon is engagement of the pro-apoptotic molecule Fas by its ligand (FasL), which leads to apoptosis, and consequently limits an inflammatory response. This idea has recently been challenged, and here we review the evidence for and against a role for FasL in immune privilege. PMID- 11733772 TI - Alfred Nobel--his life and work. AB - December 2001 marks 100 years since the first Nobel prizes were awarded. But who was Alfred Nobel, and how and why were the prizes that bear his name established? PMID- 11733773 TI - Life scientists and politics in the United States. AB - Many biologists are active in politics in the United States. Their efforts combined with those of other advocates, such as patient groups and members of Congress, have made funding of biomedical research a priority for both of the main political parties. Life scientists, acting through scientific societies and the National Academies of Science, also have a strong influence on political debates about issues with a scientific basis. PMID- 11733774 TI - Lessons in science politics. AB - Today, biology has become a big science, which devours huge amounts of money. In the United States, a phenomenal growth in biomedical science funding has paralleled the increase in research costs. But this did not occur spontaneously, and the American research community has had to learn how to influence budget policy. In Europe, we are now also faced with a similar problem and need to learn how to campaign for better research funding. PMID- 11733775 TI - Influencing science policy in Japan. AB - The administrative structure for the funding of science and technology in Japan is now going through a marked change. The newly formed Council for Science and Technology Policy (CSTP) has been given great powers - but how, in particular, is the funding policy being addressed by the new administrative structure, and how can scientists influence science policy in Japan? PMID- 11733793 TI - BAC to the future: the use of bac transgenic mice for neuroscience research. PMID- 11733794 TI - The influence of environment and experience on neural grafts. PMID- 11733795 TI - Dendritic spines: structure, dynamics and regulation. PMID- 11733796 TI - Localization and translation of mRNA in dendrites and axons. PMID- 11733797 TI - Molecules, maps and synapse specificity. PMID- 11733798 TI - Reconstructing functional systems after lesions of cerebral cortex. PMID- 11733799 TI - Visual illusions and neurobiology. PMID- 11733801 TI - Use of oral antihypertensive medication preceding blood pressure elevation in hospitalized patients. AB - OBJECTIVE: To evaluate the frequency of oral antihypertensive medication preceding the increase in blood pressure in patients in a university hospital, the drug of choice, and the maintained use of antihypertensive medication. METHODS: Data from January to June 1997 from the University Hospital Professor Edgard Santos Pharmacy concerning the prescriptions of all inpatients were used. Variables included in the analysis were: antihypertensive medication prescription preceding increase in blood pressure, type of antihypertensive medication, gender, clinical or surgical wards, and the presence of maintained antihypertensive medication. RESULTS: The hospital admitted 2,532 patients, 1,468 in surgical wards and 818 in medical wards. Antihypertensive medication prescription preceding pressure increase was observed in 578 patients (22.8%). Nifedipine was used in 553 (95.7%) and captopril in 25 (4.3%). In 50.7% of patients, prescription of antihypertensive medication was not associated with maintained antihypertensive medication. Prescription of antihypertensive drugs preceding elevation of blood pressure was significantly (p<0.001) more frequent on the surgical floor (27.5%; 405/1468) than on the medical floor (14.3%; 117/818). The frequency of prescription of antihypertensive drugs preceding elevation of blood pressure without maintained antihypertensive drugs and the ratio between the number of prescriptions of nifedipine and captopril were greater in surgical wards. CONCLUSION: The use of antihypertensive medication, preceding elevation of blood pressure (22.8%) observed in admitted patients is not supported by scientific evidence. The high frequency of this practice may be even greater in nonuniversity hospitals. PMID- 11733800 TI - Influence of ischemic preconditioning in myocardial protection in patients undergoing myocardial revascularization with intermittent crossclamping of the aorta. Analysis of ions and blood gases. AB - OBJECTIVE: To test the hypothesis that short periods of ischemia may increase the myocardial protection obtained with intermittent crossclamping of the aorta. METHODS: In the control group (18 patients), surgery was performed with systemic hypothermia at 32 degrees C and intermittent crossclamping of the aorta. Extracorporeal circulation was used. In the preconditioning group (17 patients), 2 crossclampings of the aorta lasting 3min each were added prior to the intermittent crossclamping of the conventional technique with an interval of 2min of reperfusion between them. Blood samples for analyses of pH, pCO2, pO2, sodium, potassium, calcium, and magnesium were obtained from the coronary sinus at the beginning of extracorporeal circulation (time 1), at the end of the first anastomosis (time 2), and at the end of extracorporeal circulation (time 3). RESULTS: No difference was observed in the results of the 2 groups, except for a variation in the ionic values in the different times of blood withdrawal; sodium values, however, remained stable. All patients had a good clinical outcome. CONCLUSION: The results of intermittent crossclamping of the aorta with moderate hypothermia were not altered by the use of ischemic preconditioning. PMID- 11733802 TI - Food consumption of adolescents. A simplified questionnaire for evaluating cardiovascular risk. AB - OBJECTIVE: To develop a simplified questionnaire for self-evaluation by adolescents of foods associated with the risk of coronary diseases. METHODS: Frequency questionnaires about 80 foods were answered by representative samples of 256 adolescents aged 12 to 19 from Rio de Janeiro as part of the Nutrition and Health Research project. The dependent variable was the serum cholesterol predicting equation as influenced by diet, and the independent variables were the foods. The variables were normalized and, using Pearson's correlation coefficient, those with r>0.10 were selected for the regression model. The model was analyzed for sex, age, random sample, and total calories. Those food products that explained 85% of the cholesterol variation equation were present in the caloric model, and contained trans fatty acids were selected for the questionnaire. RESULTS: Sixty-five food products had a statistically significant correlation (P<0.001) with the dependent variable. The simplified questionnaire included 9 food products present in all tested models: steak or broiled meat, hamburger, full-fat cheese, French fries or potato chips, whole milk, pies or cakes, cookies, sausages, butter or margarine. The limit of the added food points for self-evaluation was 100, and over 120 points was considered excessive. CONCLUSION: The scores given to the food products and the criteria for the evaluation of the consumption limits enabled the adolescents to get to know and to balance their intake. PMID- 11733803 TI - Some predictors of cardiovascular mortality among the elderly population of Botucatu (SP). AB - OBJECTIVE: To detect factors associated with cardiovascular mortality in the elderly of Botucatu. METHODS: We evaluated 29 variables of interest in a cohort of patients aged 60 using data from a survey conducted between 1983/84. The elderly cohort was analyzed in 1992 to detect the occurrence of cardiovascular deaths. Survival analysis was performed using the Kaplan-Meier method, the log rank test, and Cox regression analysis. Three models were adapted for each group of variables, and a final model was chosen from those variables selected from each group. RESULTS: We identified predictor for cardiovascular death according to age for elderly males not supporting the family, not possessing a vehicle, and previous cardiovascular disease. In elderly females, the predictor variables were previous cardiovascular disease and diabetes mellitus. CONCLUSION: Socioeconomic indicators (family heading and vehicle ownership) may be added to well stabilished medical factors (diabete mellitus and hypertension to select target groups for programs intended to reduce deaths due to cardiovascular diseases in elderly people. PMID- 11733804 TI - Corrected transposition of the great arteries with several associated anomalies in a 68-year-old patient. AB - Few patients with corrected transposition of the great arteries survive past 50 years of age because of the association with congenital defects, development of total atrioventricular block, and right ventricular dysfunction. We report the case of a male patient with dextrocardia in situs solitus and corrected transposition of the great arteries associated with a wide atrial septal defect and severe pulmonary valvar and subvalvar stenoses. The patient also developed a large aneurysm on the pulmonary artery, total atrioventricular block diagnosed 8 years earlier, symptoms of dysfunction of the systemic ventricle in the previous 2 years, insufficiency of the left atrioventricular valve, and aortic regurgitation. Despite all these associated anomalies, the patient developed class III cardiac decompensation only at the age of 68 years, which makes this case a rarity. The patient was clinically treated, and was discharged from the hospital in good condition. PMID- 11733805 TI - Lipoma of the right atrium. AB - The patient is a 54-year-old asymptomatic male with a tumor in the right atrium that was diagnosed on transesophageal echocardiography and confirmed as a lipoma of the right atrium on computerized tomography. The patient underwent surgical repair with extracorporeal circulation. The tumor was resected, and its base of implantation in the atrium was repaired with a flap of bovine pericardium. The diagnosis of lipoma was confirmed on histopathological examination. Locating of the tumor with the aid of transesophageal echocardiography was very useful in the strategy of cannulation of the venae cava for installation of the circuit of extracorporeal circulation. The patient had a good postoperative evolution. PMID- 11733806 TI - Clinicopathologic session. Case 5/2001 - Heart failure and insufficiency of the aortic and mitral valves in a 68-year-old woman with rheumatoid arthritis. PMID- 11733807 TI - [Clinico-radiographic correlation. Case 10/2001 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 11733808 TI - The surgeon. PMID- 11733809 TI - [The most urgent need: to acquire our own experience]. PMID- 11733811 TI - [III Guidelines for the Use of Ambulatory Blood Pressure Monitoring - ambulatory monitoring of blood pressure. Sociedade Brasileira de Hipertensao]. PMID- 11733812 TI - [I Guidelines for the Use of Home Blood Pressure Monitoring. Sociedade Brasileira de Hipertensao]. PMID- 11733813 TI - Mechanical versus biological aortic valve implants in the elderly. A comparison of early and mid-term results. AB - OBJECTIVE: Our aim was to compare, in a non randomized study, the surgical outcome in elderly patients with mechanical (Group 1; n=83) and bioprosthetic valve implants (Group 2; n=136). METHODS: During a three year period, 219 patients >75 years underwent Aortic Valve Replacement. The groups matched according to age, sex, comorbidity, valve pathology and concomitant Coronary Artery Bypass Surgery. Follow-up was a total of 469 patient-years (mean follow-up 2.1 years, maximum 4,4 years). RESULTS: Operative mortality was zero and the overall early mortality was 2.3 % (within 30 days). Actuarial survival was 87.5 +/- 4.0% and 66.1 +/- 7.7% (NS) at 4 years in Group 1 and Group 2, respectively. Freedom from valve-related death was 88.9 +/- 3.8% in Group 1 and 69.9+/-7.9% (NS) in Group 2 at 4 years. CONCLUSION: Aortic Valve Replacement in the elderly (>75 years) is a safe procedure even in cases where concomitant coronary artery revascularization is performed. Only a few anticoagulant-related complications were reported and this may indicate that selected groups of elderly patients with significant life expectancy may benefit from mechanical implants. PMID- 11733814 TI - Video-assisted pericardioscopy. How to improve diagnostic efficacy in pericardial effusions. AB - OBJECTIVE: To assess, in a prospective way, the experience with video-assisted pericardioscopy obtained in patients with pericardial effusion of unclear etiology in the preoperative period. METHODS: From January 1998 to June 2000, 20 patients were operated upon with the aid of video-assisted pericardioscopy. On echocardiography, 17 of these patients had significant pericardial effusion, and 3 had moderate pericardial effusion. Video-assisted pericardioscopy was performed through a small incision of the Marfan type. RESULTS: The diagnosis of pericardial effusion was established as follows: idiopathic in 9 (45%) patients, neoplastic in 4 (20%), resulting from hypothyroidism in 3 (15%), tuberculous in 2 (10%), due to cholesterol in 1 (5%), and chylopericardial in 1 (5%). The biopsy was positive in 30% of the patients, and the etiology could not be defined in 45% of the patients. CONCLUSION: Video-assisted pericardioscopy proved to be a method with low morbidity and a high index of diagnostic positivity. A high percentage of pericardial effusions are caused by viral infections, which are not diagnosed through current methods, being, therefore, classified as idiopathic. PMID- 11733815 TI - Treatment of atrial fibrillation with radiofrequency ablation and simultaneous multipolar mapping of the pulmonary veins. AB - OBJECTIVE: To demonstrate the feasibility and safety of simultaneous catheterization and mapping of the 4 pulmonary veins for ablation of atrial fibrillation. METHODS: Ten patients, 8 with paroxysmal atrial fibrillation and 2 with persistent atrial fibrillation, refractory to at least 2 antiarrhythmic drugs and without structural cardiopathy, were consecutively studied. Through the transseptal insertion of 2 long sheaths, 4 pulmonary veins were simultaneously catheterized with octapolar microcatheters. After identification of arrhythmogenic foci radiofrequency was applied under angiographic or ultrasonographic control. RESULTS: During 17 procedures, 40 pulmonary veins were mapped, 16 of which had local ectopic activity, related or not with the triggering of atrial fibrillation paroxysms. At the end of each procedure, suppression of arrhythmias was obtained in 8 patients, and elimination of pulmonary vein potentials was accomplished in 4. During the clinical follow-up of 9.6+/-3 months, 7 patients remained in sinus rhythm, 5 of whom were using antiarrhythmic drugs that had previously been ineffective. None of the patients had pulmonary hypertension or evidence of stenosis in the pulmonary veins. CONCLUSION: Selective and simultaneous catheterization of the 4 pulmonary veins with microcatheters for simultaneous recording of their electrical activity is a feasible and safe procedure that may help ablation of atrial fibrillation. PMID- 11733816 TI - Atrial activation during sinus rhythm in patients with rheumatic and non rheumatic paroxysmal atrial fibrillation. Frequency-domain analysis using signal averaged electrocardiography. AB - OBJECTIVE: Using P-wave signal-averaged electrocardiography, we assessed the patterns of atrial electrical activation in patients with idiopathic atrial fibrillation as compared with patterns in patients with atrial fibrillation associated with structural heart disease. METHODS: Eighty patients with recurrent paroxysmal atrial fibrillation were divided into 3 groups as follows: group I - 40 patients with atrial fibrillation associated with non-rheumatic heart disease; group II - 25 patients with rheumatic atrial fibrillation; and group III - 15 patients with idiopathic atrial fibrillation. All patients underwent P-wave signal-averaged electrocardiography for frequency-domain analysis using spectrotemporal mapping and statistical techniques for detecting and quantifying intraatrial conduction disturbances. RESULTS: We observed an important fragmentation in atrial electrical conduction in 27% of the patients in group I, 64% of the patients in group II, and 67% of the patients in group III (p=0.003). CONCLUSION: Idiopathic atrial fibrillation has important intraatrial conduction disturbances. These alterations are similar to those observed in individuals with rheumatic atrial fibrillation, suggesting the existence of some degree of structural involvement of the atrial myocardium that cannot be detected with conventional electrocardiography and echocardiography. PMID- 11733817 TI - The impact of syncope during clinical presentation of sustained ventricular tachycardia on total and cardiac mortality in patients with chronic Chagasic heart disease. AB - OBJECTIVE: To assess the impact of syncope during sustained ventricular tachycardia on total and cardiac mortality in patients with chronic chagasic heart disease. METHODS: We assessed 78 patients with sustained ventricular tachycardia and chronic Chagas' heart disease. The mean age was 53+/-10 years, 45 were males, and the mean ejection fraction was 49.6+/-13%. The patients were divided into 2 groups according to the presence (GI=45) or absence (GII=33) of syncope during sustained ventricular tachycardia. RESULTS: After a mean follow-up of 49 months, total mortality was 35% (28 deaths), 22 deaths having a cardiac cause (78.6%). No difference was observed in total (33.3% x 39.4%) and cardiac (26.7% x 30.3%) mortality, or in nonfatal sustained ventricular tachycardia between GI and GII patients (57.6% x 54.4%, respectively). However, the presence of syncope during recurrences was significantly greater in those patients who had had the symptom from the beginning (65.4% x 18.1%, p<0.01). CONCLUSION: Syncope during the presentation of sustained ventricular tachycardia is not associated with an increase in total or cardiac mortality in patients with chronic Chagas' heart disease. However, syncope during the recurrence ventricular tachycardia is greater in patients experiencing syncope in the first episode, of sustained ventricular tachycardia. PMID- 11733818 TI - The Bambui health and aging study (BHAS). Prevalence of intermittent claudication in the aged population of the community of Bambui and its associated factors. AB - OBJECTIVE: To assess the prevalence of intermittent claudication in the aged population of Bambui, Brazil, and to identify the factors associated with this disease. METHODS: Population-based cross-sectional study of the aged population (>/= 60 years of age) of Bambui. Participants were interviewed and examined, after written consent. Intermittent claudication was defined based on a standardized questionnaire. Analysis was performed using multiple logistic regression. RESULTS: Of the 1,742 elderly living in Bambui, 1,485 (85.2%) were enrolled in the study. Thirty-seven individuals (2.5%) with intermittent claudication were identified: 28 (1.9%) males and 9 (0.6%) females. Their age brackets were: 16 (1.08%) individuals between 60 and 69 years of age, 17 (1.15%) between 70 and 79 years, and 4 (0.27%) >/= 80 years. A significant association between intermittent claudication and the following characteristics was found: male sex (OR=5.1; CI 2.4-11.0), smokers (OR=3.1; CI 1.2-8.5), ex-smokers (OR=3.4; CI 1.3-8.7), and more than 2 hospital admissions in the last 12 months (OR=2.8; CI 1.1-7.2). CONCLUSION: Disease prevalence was similar to that of other countries. The association between intermittent claudication and smoking strengthens the significance of tobacco in peripheral artery disease pathogenesis. The association of intermittent claudication and a higher number of hospital admissions suggests greater morbidity in the elderly affected. PMID- 11733819 TI - [Manidipine in the treatment of stage I and II essential hypertension patients with overweight or android obesity. A Brazilian multicentre study of efficacy, tolerability and metabolic effects]. AB - OBJECTIVE: To evaluate the efficacy, metabolic effects and tolerability of manidipine used in the treatment of stage I and II essential hypertensive patients with overweight or android obesity. METHODS: By an open-label, non comparative protocol in 11 Brazilian clinical research centers 102 hypertensive patients of both sexes with over weight or central obesity were treated with manidipine 10 to 20mg once daily for 12 weeks. Blood pressure, heart rate and adverse events were monitored. Fasting plasma glucose, total, HDL and LDL cholesterol and triglicerides were determined at both placebo period and end of active treatment. Also in 12 patients, insulin sensitivity index was evaluated during placebo and manidipine treatment. RESULTS: Blood pressure was reduced from 159+/-15 / 102+/-5mmHg to 141+/-15 / 90+/-8mmHg with the treatment without any noticeable change in heart rate. Manidipine-efficacy rate was 71.9% with 51.1% of blood pressure normalization. No significant changes in metabolic parameters were noticed. Tolerability to manidipine was very high and at the last visit 87.1% of the treated patients were free of any adverse event. CONCLUSION: Manidipine is an adequate, highly effective, exempt of metabolic effects and safe option for treatment of stage I and II essential hypertensive patients with overweight or android obesity. PMID- 11733820 TI - New percutaneous techniques for perforating the pulmonary valve in pulmonary atresia with intact ventricular septum. AB - We report new percutaneous techniques for perforating the pulmonary valve in pulmonary atresia with intact ventricular septum, in 3 newborns who had this birth defect. There was mild to moderate hypoplastic right ventricle, a patent infundibulum, and no coronary-cavitary communications. We succeeded in all cases, and no complications related to the procedure occurred. The new coaxial radiofrequency system was easy to handle, which simplified the procedure. Two patients required an additional source of pulmonary flow (Blalock-Taussig shunt) in the first week after catheterization. All patients had a satisfactory short term clinical evolution and will undergo recatheterization within 1 year to define the next therapeutic strategy. We conclude that this technique may be safely and efficiently performed, especially when the new coaxial radiofrequency system is used, and it may become the initial treatment of choice in select neonates with pulmonary atresia and intact ventricular septum. PMID- 11733821 TI - Aortopulmonary window and double aortic arch. A rare association. AB - We report the case of a 27-year-old male patient with dyspnea on physical exertion. Clinical assessment and various tests led to the diagnosis of aortopulmonary window and double aortic arch. According to a literature search, this may be the first report on such association. PMID- 11733822 TI - [Clinical-radiographic correlation. Case 11/2001 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 11733823 TI - Management of mild aortic stenosis in patients undergoing coronary bypass surgery. PMID- 11733824 TI - Diffusion-weighted MR imaging of cystic lesions of neurocysticercosis: a preliminary study. AB - Neurocysticercosis is an endemic disease in some developing countries. It has pleomorfic clinical and imaging findings, which are variable from patient to patient. In this preliminary note, we studied the magnetic resonance (MR) diffusion-weighted images (DWI) of sixteen patients presenting with cystic lesions of this disease diagnosed by clinical and laboratorial findings. All the lesions had hypointense signal and the similar apparent diffusion coeficient (ADC) values as the cerebrospinal fluid (CSF). PMID- 11733825 TI - Dosage of lactate in the cerebrospinal fluid in infectious diseases of the central nervous system. AB - This paper analyzes the diagnosis aid of the dosage of lactate in the cerebrospinal fluid (CSF) in infectious diseases of the central nervous system (CNS). We analyzed prospectively 130 samples of CSF of 116 patients with diagnoses of infectious processes in the CNS. The 130 samples of CSF were divided into five groups: 28 samples of the control group, 40 of bacterial meningitis, 22 of viral meningitis, 16 of fungal meningitis and 24 of patients presenting acquired immune deficiency syndrome (AIDS). The concentration of lactate in the CSF was elevated in the group of patients with bacterial meningitis (average = 46.2 mg/dL), fungal meningitis (average = 27.3 mg/dL) and in the AIDS group (average = 23.5 mg/dL). In the control group and viral meningitis group the lactate content in the CSF presented the reference rates according to the employed method. The lactate dosage in the CSF presented a negative correlation with glycorrhachia and positive correlation with the cellularity and total proteins of the CSF. We conclude that the lactate dosage in the CSF, although unspecific, helps to distinguish the infectious processes of the CNS. PMID- 11733826 TI - Neurological evaluation of children and adolescents with brain tumor, based on ambulatory-oriented follow-up. AB - Taken as proved that brain tumors are the second most frequent childhood neoplasm - only outnumbered by leukemias - we have undertaken a clinical perspective study with seventy brain tumor patients ranging from one to fifteen years of age, throughout a four-year period (1993-1997), based on ambulatory-oriented follow up. Forty-one male and twenty-nine female patients were analyzed, in that a slightly higher number of infratentorial tumors was observed (thirty-eight cases), compared to those supratentorially located (thirty-two cases). The most repeatedly observed during the study was the medulloblastoma (twenty-one patients), followed by the astrocytoma (fifteen patients) and the germinoma (eleven patients). It should be pointed out that during the ambulatory follow-up 75,5% of patients developed neurological sequels. A tumor recurrence was noticed in 34,3% of them, while 21,4% eventually died. PMID- 11733827 TI - Identification of family variables in parents' groups of children with epilepsy. AB - OBJECTIVE: To verify the effectiveness of the support group in the identification of family variables linked to epilepsy. METHOD: Pre-test were applied to parents of 21 children with benign epilepsy of childhood recently diagnosed, from 5 to 15 years, who participated in the groups at HC/Unicamp. There was a presentation of an educational video, discussion and application of the post-test 1. After six months, the post-test 2 was applied. RESULTS: The beliefs were: fear of swallowing the tongue during the seizures (76.19%) and of a future mental disease (66.67%). Facing the epilepsy, fear and sadness appeared. 76.19% of the parents presented overprotection and 90.48%, expected a new seizure. In the post-test 1, the parents affirmed that the information offered had modified the beliefs. In the post-test 2, 80.95% didn't report great doubts about epilepsy and 90.48% considered their relationship with their children better. CONCLUSIONS: The demystification of beliefs supplied from the groups influenced the family positively, prevented behavior alterations and guaranteed effective care in the attendance to the child with epilepsy. PMID- 11733828 TI - Efficacy and safety of risperidone oral solution in agitation associated with dementia in the elderly. AB - BACKGROUND: Behavioral and psychological symptoms in dementia (BPSD) contribute to caregiver burden and institutionalization of elderly. Neuroleptics are prescribed to control agitation. Side effects of typical neuroleptics are harmful, making atypical neuroleptics an indication. OBJECTIVES: To evaluate efficacy and tolerability of risperidone oral solution (ROS) given once daily to demented elderly outpatients with BPSD (agitation). METHOD: Patients (n=26), 76.35+/-8.63 years, Diagnostic and Statistical Manual of Mental Disorders 4th ed. (DSM-IV) criteria for dementia. RSO was given, starting dose of 0.25 mg and increments of 0.25 mg every week. Mini-Mental State Examination (MMSE) assessed cognitive status, Behavioral and Emotional Activities Manifested in Dementia (BEAM-D) and Clinical Global Impression (CGI) measured BPSD, Extrapiramidal Symptom Rating Scale (ESRS) evaluated extrapyramidal symptoms. Cardiovascular side effects were evaluated clinically. RESULTS: There was a 26% reduction in agitation and no cardiovascular side effects in the range from 1.0 to 1.25 mg. Side effects were more prevalent above 2.5 mg. CONCLUSION: Risperidone oral solution improved agitation with good tolerability from 0.5 to 1.25 mg. A single dose with increments of 0.25 mg may be more acceptable to patients and caregivers. PMID- 11733829 TI - [Alzheimer's disease and magnetic resonance spectroscopy of the hippocampus]. AB - OBJECTIVE: Acquisition of data of magnetic resonance metabolite spectrum of the hippocampal formation (hippocampus-hc) in the elderly, normal and with Alzheimer's disease (AD). METHOD: Subjects matched for age: a. normal sample (n=20), CDR=0, and b. AD sample (n=40), CDR 1 and 2. TECHNIQUE: Signa Horizon LX GE, 1.5T, 1H-MRS with automated software PROBE/SV, VOI: hc (right and left); single voxel (2x2x2cm); TR 1500ms/TE 50ms; PRESS; metabolites: N-acetylaspartate (Naa), choline (Cho), creatine (Cr), myo-inositol (mI). RESULTS: The present data relate to the ratios of Naa, Cho and mI, with Cr taken as reference, and the mI/Naa ratio. The study showed reduction of Naa, increase of mI and of the mI/Naa ratio, and not consistent results for Cho. The results of the whole sample of AD patients compared to the pooled normal mean +/- sd were significant for Naa, mI and mI/Naa (p<0.01). Accuracy in relation to the individual values of both samples showed satisfactory levels of sensitivity, specificity and positive predictive value. CONCLUSION: The present results can be used as a helpful tool to detect pathologic changes of the hippocampus in AD, and allowing greater accuracy and an earlier diagnosis of this disease. PMID- 11733830 TI - [Does semi-quantitative analysis of FDG-PET have any additional value in the diagnosis of mesial temporal sclerosis?]. AB - The aim of this study was to compare qualitative visual analysis with semi quantitative analysis in the diagnosis of mesial temporal sclerosis (MTS) using FDG-PET. METHODS: This study included 21 patients with histopathological confirmation of MTS. FDG-PET visual analysis data were based on clinical reports generated soon after the completion of the scan. FDG-PET images were semi quantitatively analyzed using regions of interest (ROIs) in 19 slices perpendicular to the longest axis of the temporal lobe. These ROIs divided each temporal lobe into three regions (lateral, inferior and medial). An asymmetry index was calculated for each region. RESULTS: The visual analysis of the FDG-PET studies demonstrated asymmetric hypometabolism in all patients. All but 1 patient underwent standard lobectomy of the same side described as hypometabolic by the PET report. Using an asymmetry index equal or greater than 9% in at least one of the regions as a threshold, the FDG-PET semi-quantitative analysis showed significant asymmetry in 18 patients. These also matched the side of lobectomy and were confirmed as sclerotic in all but one patient (same patient as above). CONCLUSION: The semi-quantitative analysis did not provide additional information over visual interpretation in this series of patients. PMID- 11733831 TI - [Eletroencephalographic ambulatory monitoring in refractory epilepsies in childhood]. AB - The objective of our study was, by means of continuous prolonged ambulatory electroencephalographic monitoring, to analyze the temporal distribution of paroxysmal discharges during sleep and awake in children and adolescents with refractory epilepsies. Twenty-one patients in the 4-to-17 year age bracket with refractory epilepsies, with 52.3% (n=11) male and 47.6% (n=10) female from the Discipline of Neurology of the Universidade Federal de Sao Paulo (Federal University of Sao Paulo). Cerebral Holter was carried out with Bioware EEG-2008 of prolonged ambulatory electroencephalographic monitoring equipment. We observed greater frequency of isolated and grouped epileptic discharges in day and in night sleep in relation to awake; day and night sleep led to activation of epileptic discharges, both isolated and grouped. The cerebral Holter was more effective in detecting epileptiform discharges than the routine EEG in 33.33% of the patients. The cerebral Holter proved a useful and precise method in detecting epileptic discharges, as an aid in the assessment of the fluctuations in frequency of paroxysmal activity in children with refractory epilepsies, both in relation to activities in daily life, and to the relation to the biological cycle of sleep and awake. PMID- 11733832 TI - [Attention deficit hyperactivity disorder: proposal of neuropsychological assessment]. AB - Attention deficit hyperactivity disorder (ADHD) is characterized by a persistent pattern of inattention and/or hyperactivity/impulsivity. The aim of this research was to contribute more precisely to the diagnosis of ADHD, to propose a battery of neuropsychological assessment and to analyze the contribution of each test. We studied 10 matched pairs of children with ADHD and normal controls (7 to 11 years). Inclusion criteria were: presence of ADHD typical behavior, positive diagnosis of ADHD based on DSM-IV, normal IQ, normal neurological examination and parental consent. We used extensive neuropsychological battery. The results showed differential sensitivity for detection of attentional problems in children with ADHD, although most tests did not reach statistical significance. The item, errors, of WCST revealed statistically significant difference between the two groups: ADHD performance was inferior to controls. In conclusion the neuropsychological assessment battery used in this research contributed to the diagnosis of ADHD. PMID- 11733833 TI - [Box and block test of manual dexterity in normal subjects and in patients with multiple sclerosis]. AB - Recently new disease-modifying treatments for multiple sclerosis (MS) were introduced which can change the natural course of the disease. In clinical trials with these new agents the Expanded Disability Status Scale (EDSS) is often used as a primary outcome instrument to measure neurological impairment and disability. A number of limitations have been identified when using the EDSS, some of wich are because the EDSS is an ordinal scale that is heavily biased to locomotor function. In this study we applied the box and block test of manual dexterity in normal subjects and relapsing-remitting MS patients. The results were that 64.8% of the female and 80.7% of the male patients had significant changes on this task compared with normal subjects, and as this test is easily applied and is sensitive in detecting upper extremity functional ability, we recommend its use in clinical trials to evaluate new drugs in MS patients. PMID- 11733834 TI - [Technical aspects of intracranial pressure monitoring by subarachnoid method in severe head injury]. AB - Two hundred and six patients with severe head injury (Glasgow Coma Scale of 8 points or less after nonsurgical resuscitation on admission), managed at Intensive Care Unit-Hospital das Clinicas - Universidade Estadual de Campinas were prospectively analysed. All patients were assessed by CT scan and 72 required neurosurgical intervention. All patients were continuously monitored to evaluate intracranial pressure (ICP) levels by a subarachnoid device (11 with subarachnoid metallic bolts and 195 with subarachnoid polyvinyl catheters). The ICP levels were continuously observed in the bedside pressure monitor display and their end-hour values were recorded in a standard chart. The patients were managed according to a standard protocol guided by the ICP levels. There were no intracranial haemorrhagic complications or hematomas due the monitoring method. Sixty six patients were punctured by lateral C1-C2 technique to assess infectious complications and 2 had positive cerebrospinal fluid samples for Acinetobacter sp. The final results measured at hospital discharge showed 75 deaths (36,40%) and 131 (63,60%) survivors. ICP levels had significantly influenced the final results (p<0,001). The subarachnoid method to continuously assess the ICP levels was considered applicable, safe, simple, low cost and useful to advise the management of the patients. The ICP record methodology was practical and useful. Despite the current technical advances the subarachnoid method was considered viable to assess the ICP levels in severe head injury. PMID- 11733835 TI - [Neuroendoscopic endonasal transseptal approach to the sellar region: study of 30 cases]. AB - An endoscopic endonasal transsphenoidal approach to the sella was performed in 30 patients. The use of endoscope allowed close inspection and differentiation between tumor tissue and glandular remains, resulting in micro dissection of the tumor with maximum preservation of pituitary function. The angled view of the endoscope helps gross total removal of the tumor tissue, from the supra and para sellar extension. The endonasal transseptal endoscopic approach helps from dental problems, and lowers incidence of septal perforation, synechia, and crust formation. However the endoscopic approach has drawbacks. Its decreases the depth of field, needs constant manual control of the endoscope, and requires experience with the endoscope technique. PMID- 11733836 TI - [Hormonal dysfunction of nonpituitary lesions from midline and perisellar area]. AB - OBJECTIVE: To analyse clinical and/or laboratorial preoperative hormonal dysfunction, of the nonpituitary intracranial lesions from midline and parasellar area. METHOD: Forty-four patients were evaluated with nonpituitary intracranial lesions, who had images studies (computed tomography or magnetic resonance) and preoperative basal hormonal level; 16 had preoperative hypothalamus-hypophysial function tests (megatests). These patients were divided in two groups. Group I - 34 lesions from midline: 11 craniopharyngiomas, 8 meningiomas, 3 germinomas, 3 tumors of sphenoid sinus, 2 empty sella syndrome, 2 pylocitic astrocytomas, 1 giant aneurysm, 2 mucoceles, 1 III ventricle diverticulum and 1 Rathke's cleft cyst; Group II - 10 lesions from parasellar area: 9 meningiomas and 1 giant aneurysm. RESULTS: In group I, 25/34 (73.5%) patients showed laboratorial hormonal deficit (14 without clinical manifestations) 18/34 (52.9%) hyperprolactinemia (5 with galactorreia) and 8 (53.3%) showed growth hormone deficiency in 15 megatests available in this group; 3 (8.8 %) patients presented central diabetes insipidus (CDI). In group II, 6/10 (60%) patients showed laboratorial hormonal deficit (5 without clinical manifestations), 1 (10%) hyperprolactinemia and 1 growth hormone deficiency (single megatest realized in this group); no patient had preoperative CID. CONCLUSIONS: The presence of nonspecific or poorly valorized clinical manifestations, does not indicate absence of hormonal dysfunction; in this present serie, 19/38 (50%) patients with laboratorial abnormalities, didn't show clinical manifestations. Hormonal dysfunction is frequent in sellar and perisellar nonpituitary lesions, specially involving midline. PMID- 11733837 TI - [Holoprosencephaly: morphological aspects of twelve cases of autopsy]. AB - The term holoprosencephaly (HPC) is used to indicate the group of hemispheric deformities caused by a failure in the development of the prosencephalic vesicle. The purpose of this study is to explain the morphologic spector of twelve cases of HPC, qualify them, and compare them to the literature. It was evaluated 5837 pediatrics necropsies, and there were 12 cases of HPC. Data like gender, age and the presence of another associated malformations were evaluated and compared to another malformations and to the total number of necropsies. The majority of the cases was male (66.66%), and stillborns (75%). The most frequently type of HPC found was the lobar type (58.3%), and the most frequently type of facial alteration was the ciclopy (25%). There is a statistic tendency to HPC affect the male sex, in comparison with other neurologic malformations. Besides, the stillborns are more frequently observed in HPC than in other neurologic malformations. PMID- 11733838 TI - [Cerebrovascular diseases in patients aged 15 to 40 years: neuropathological findings in 47 cases]. AB - Cerebrovascular diseases in patients between 15 and 40 years old are not a frequent subject in Latin-American literature, especially when focusing on neuropathology. We analyzed 47 brains from necropsies performed from 1987 to 1997 and selected on a basis of age and the presence of vascular pathology. From the 47 analyzed brains, 26 belonged to females (55.3%). When distributed among age groups, 12.8% (n=6) affected patients from 15 to 20 yo, 51,1% (n=24) from 21 to 30 yo, 36,2% (n=17) from 31 to 40 yo. The underlying diseases were: cardiac and haemathologic (19.2%), pregnancy complications (12.76%), infections, diseases of blood vessels and neurological (10.1% each) amongst others. The neuropathological abnormalities included cerebral and/or cerebellar herniation (16%), cerebral edema (13.8%), subarachnoid hemorrhage (10%), recent cerebral infarction (9%), intraparenchymatous hemorrhage (8.14%), hypoxic-ischemic encephalopathy (3%) and other events such as Sneddon syndrome and Lupus vasculitis. These findings express that the cerebrovascular phenomena in this age group are unique and closely related with the underlying disease. PMID- 11733839 TI - [Fraction of gemistocytic astrocytes and immunoexpression of p53 protein in astrocytomas grade II and III (WHO)]. AB - Twenty-two patients with astrocytomas, grade II or III WHO, were studied from 1990 to 1998. In all cases, histopathology showed that the astrocytomas had a gemistocytic component. The aims of this study were to establish the fraction of gemistocytic astrocytes, to investigate p53 protein immunoexpression and to evaluate correlations between these two parameters with the tumour outcome. Tumor cells were quantified at high-power magnification (x400). At least 1000 neoplastic cells (small neoplastic astrocytes plus gemistocytes) were counted in each specimen. The percentage of gemistocytes was defined as the gemistocytic index. Nuclear expression of p53 protein was evaluated in neoplastic astrocytes and gemistocytes. Both the frequency (7/22) as well the p53 immunoexpression indices in gemistocytes, regardless of the grade of the astrocytomas, were inferior from those reported in the literature. No correlation was found between the gemistocytic indices and the p53 immunoexpression. PMID- 11733840 TI - Familial Creutzfeldt-Jakob disease associated with a point mutation at codon 210 of the prion protein gene. AB - Creutzfeldt-Jakob disease (CJD), the most known human prion disease, is usually sporadic but approximately 15% of the cases are familial. To date, seven CJD cases with codon 210 mutation (GTT to ATT) have been reported in the literature. We describe a case of a 57 year-old woman who presented gait disturbances and rapidly progressive dementia, leading to death four months after onset. Electroencephalogram revealed periodic activity, diffusion-weighted magnetic resonance imaging showed hypersignal in basal ganglia, and test for 14-3-3 protein was strongly positive in the CSF. The complete prion protein gene coding region was sequenced after PCR amplification, showing a point mutation in codon 210. This is the first case of CJD with codon 210 mutation diagnosed in Brazil. We emphasize the role of genetic search for prion protein gene mutation, even in patients presenting clinical features resembling sporadic CJD. PMID- 11733841 TI - Thrombolytic therapy for acute extra-cranial artery dissection: report of two cases. AB - Extra-cranial arterial dissection accounts for 10% of strokes in young people. Information on safety of thrombolytic administration in this group is limited. The literature, however, does not favor use of thrombolytics for myocardial ischemia when peripheral arterial dissection coexists. Based on the clinical and radiological features, two patients who presented with acute stroke secondary to arterial dissection were considered for thrombolysis. One of them received intra venous recombinant tissue plasminogen activator (rtPA), and the other patient received intra-arterial rtPA. There were no post thrombolysis complications. This report supports feasibility of administering thrombolytics in acute ischemic strokes resulting from extra-cranial arterial dissection. Future larger studies are necessary to determine the efficacy, safety and long-term outcome in this patient population. PMID- 11733842 TI - Anaplastic meningioma: case report. AB - Intracranial meningiomas continue to challenge our best clinical efforts to eliminate them once discovered and deemed appropriate for treatment. Malignant meningiomas constitute 10% to 15% of all meningiomas and limited information exists regarding adjuvant treatment. The external whole brain irradiation is recommended. Traditional chemotherapy has proven ineffective; thus, new chemotherapeutic agents and new methods of delivery should be developed. Immunotherapy may be considered for patients with malignant meningiomas when all others previous treatment have failed. We report a case of anaplastic papillary meningioma. A 67-year-old man presented with partial complex seizures, headache and aggressiveness. A computerized tomography and magnetic resonance image demonstrated a large left temporo-occipital mass with diffuse contrast enhancement and extensive surrounding edema. A left temporo-occipital flap was performed. The tumor and the infiltrated dura were radically removed. Postoperatively, the patient remained neurologically intact. The treatment was complemented by external whole brain radiation. PMID- 11733843 TI - Episodic paroxysmal hemicrania with seasonal variation: case report and the EPH cluster headache continuum hypothesis. AB - Episodic paroxysmal hemicrania (EPH) is a rare disorder characterized by frequent, daily attacks of short-lived, unilateral headache with accompanying ipsilateral autonomic features. EPH has attack periods which last weeks to months separated by remission intervals lasting months to years, however, a seasonal variation has never been reported in EPH. We report a new case of EPH with a clear seasonal pattern: a 32-year-old woman with a right-sided headache for 17 years. Pain occurred with a seasonal variation, with bouts lasting one month (usually in the first months of the year) and remission periods lasting around 11 months. During these periods she had headache from three to five times per day, lasting from 15 to 30 minutes, without any particular period preference. There were no precipitating or aggravating factors. Tearing and conjunctival injection accompanied ipsilaterally the pain. Previous treatments provided no pain relief. She completely responded to indomethacin 75 mg daily. After three years, the pain recurred with longer attack duration and was just relieved with prednisone. We also propose a new hypothesis: the EPH-cluster headache continuum. PMID- 11733844 TI - [Cerebral vasculitis and Basedow-Graves disease: report of two cases]. AB - OBJECTIVE: To report two cases of patients with cerebral vasculitis and Basedow Graves disease. CASE REPORT: Two young female patients presented at emergency with acute neurological picture of hemiparesis and speech distress. The first patient, 22 year-old, has been on clinical treatment for hyperthyroidism for one month. On clinical examination she had an elevated cardiac rate, bilateral exophthalmia and a diffuse goiter. On neurological examination we could observe a mild dysarthria and left hemiparesis. The second patient was a 15 year-old girl, who presented sudden conscious loss, aphasia and right hemiparesis. RESULTS: CT scan and MRI showed areas of cerebral infarcts. Angiographic study showed multiple vascular stenosis similar to an arteritic angiographic pattern. Extensive medical and laboratorial workout disclosed no other risk factor for stroke or for nervous system vasculitis. The antimicrosomal and antithyreoglobulin antibodies were positive and thyroidian hormones confirmed hyperthyroidism. CONCLUSION: The concurrence of cerebral arteritis and Basedow Graves' disease suggest a possible pathogenic link between Graves' disease and specific cerebral vascular disorders, possibly through a common autoimmune mechanism. PMID- 11733845 TI - [Cytomegalovirus encephalitis in an immunocompetent patient: clinical, neuropathological and ultrastructural analysis]. AB - Cytomegalovirus encephalitis is rarely diagnosed in immunocompetent patients. It is more frequent among immunocompromised hosts. We present a case of a young male patient without evidence of immunosupression who had a cytomegalovirus encephalitis. The diagnosis was made by detection of viral DNA in cerebral tissue obtained by biopsy. Neuropathologic and ultrastructural changes are described and compared with the descriptions in immunocompromised patients. PMID- 11733846 TI - [Mandibular trigeminal schwannoma: case report]. AB - We report one case of peripheral trigeminal schwannoma originated from the mandibular branch (V3) and located inside the infratemporal and pterygopalatine fossae with slight intracranial extension. The tumor was completely removed by a transmaxillary approach. The revision of the literature showed to be a very rare tumor in this location. PMID- 11733847 TI - [Creutzfeldt-Jakob disease: case report with spinal cord involvement]. AB - Creutzfeldt-Jakob disease (CJD) is the most common subacute transmissible spongiform encephalopathy. Approximately 85% of the cases are sporadic. The remaining 15% consist of genetic and iatrogenic forms. We report a sporadic form of CJD with spinal cord involvement and a clinical manifestation characterized by dementia and cerebellar syndrome, myofasciculation with absent reflexes and seizures. The two last manifestations are rare. The clinical hypothesis was probable CJD which was confirmed with autopsy and immunohistochemistry. We conclude that CJD should always be suspected when rapidly progressive dementia occurs and the absence of pyramidal or extrapyramidal signs suggest a spinal cord and/or peripheral nerve involvement. PMID- 11733848 TI - [Neurochristopathy in the differential diagnosis of newborn's apnea: case report]. AB - OBJECTIVE: to include neurocristopathy on the etiological workup of neonatal apneas and discuss the importance of polysomnography in this diagnosis. METHOD: case report and critical review of the literature. RESULTS: we report on a newborn that presented respiratory failure in the first hours of life associated to abdominal distention. Continuous ventilatory support was necessary, and the respiratory distress increased during sleep. After polysomnographic confirmation of hypoventilation the newborn was submitted to neuroradiolgic tests and colon byopsy, positive to Hirschsprung's disease. CONCLUSION: Neurocristopathy syndrome can have many different clinical expression, and sometimes the syndrome can be misdiagnosed. Polysomnography confirms central hypoventilation. This diagnosis should be considered in the newborn's persistent apnea workup. PMID- 11733849 TI - [First Brazilian consensus for the management of the acute phase of cerebral vascular accidents]. AB - The cerebrovascular diseases have a high incidence, and they cause an enormous social and economic burden. With the increasing knowledge of the pathophysiology of the ischemic insult, and the possibility of the thrombolysis in the acute phase, the management of the stroke patients is considered an emergency. This consensus is the result of a recent meeting to establish a better approach to these patients. This is the official guideline for the management of patients with acute stroke of the Brazilian Cerebrovascular Disease Society. PMID- 11733850 TI - The importance of ultramicroscopic emphysema in cigarette smoke-induced lung disease. AB - To determine the role of the alveolar pores in cigarette smoke-induced lung disease, we examined the alveolar pores of guinea pigs exposed to cigarette smoke for 12 months, and compared these data to those obtained from sham-smoked animals, correlating the data with airspace size and lung function. We found that the smoke-exposed animals had a larger mean number of pores per alveolus (p < 0.001), and the distributions of pore size and shape were significantly shifted to indicate a larger and more irregular pore configuration (p < 0.001, 01 respectively). In the smoke exposed group, there was a significant correlation of pore number with total lung capacity (TLC) (0.68 p < 0.05), RV (0.70, p < 0.05), and FEV(0.1)/FVC(-0.77, p < 0.02). No correlations were identified between pore size or shape and the lung function tests. We conclude that cigarette smoke exposure produces an increase in the number of alveolar pores, a process which we believe represents ultramicroscopic emphysema. These alterations appear to precede any increase in airspace size, and may help to explain abnormal lung function in cigarette smokers without macroscopic emphysema or small airway disease. This is the first study to clearly document an increased number of alveolar pores, with a significant number of either/or large and irregular pores, after chronic smoke exposure, but in the absence of gross emphysema. PMID- 11733851 TI - Exhaled nitric oxide: the effects of age, gender and body size. AB - Since little is known of the effects of age, gender, and body size on exhaled nitric oxide (NO) production, we have conducted a prospective study to examine these factors in a healthy nonsmoking women (mean age +/- SD 47.7 +/- 15.8, range 20-79 years). Exhaled NO was measured by an automatic chemiluminescence analyzer (Sievers NO Analyser 280) at steady expiration. Men had significantly higher exhaled NO levels than women (p = 0.001). Although exhaled NO levels did not correlate with age (r = 0.12, p = 0.17), it correlated significantly with height (r = 0.23, p = 0.02), weight (r = 0.34, p PMID- 11733852 TI - Expiratory muscle endurance in middle-aged healthy subjects. AB - To evaluate expiratory muscle endurance in middle-aged healthy subjects using incremental as well as constant expiratory loads, 14 healthy volunteers (51 +/- 16 years) were submitted to a specific endurance test, which was performed breathing against a threshold valve, and was divided into two parts. In part I, the load was progressively increased (50 g each 2 min) until task failure occurred. The mean mouth pressure generated against the highest load held for at least 60 sec was defined as the maximal expiratory sustainable pressure (Pth(max)). In part II, each subject breathed against a constant submaximal expiratory load (80% Pth(max)) until task failure occurred (expiratory endurance time or Tth(80)). Both parts of the test were repeated 24-48 h later. Progressive expiratory loading induced a linear increase in mouth expiratory pressure and the Pth(max) obtained was 141 +/- 43 cm H(2)O, representing 74 +/- 28% of the maximal expiratory pressure (PE(max)). Under constant loads, the Tth(80) was 17 +/- 9 min. At the end-point of both parts, the tension time index for expiratory muscles was dramatically increased (>0.25), and both EMG central frequency and PE(max) were decreased with no changes in maximal inspiratory pressure or inspiratory capacity. Extreme dyspnea was present in most of the subjects but no complications were observed. The endurance of expiratory muscles can be easily assessed in healthy subjects using this method, which has acceptable reproducibility and tolerance. PMID- 11733853 TI - Morphological and biochemical properties of alveolar fibroblasts in interstitial lung diseases. AB - The phenotype of alveolar-associated fibroblasts (Afb) in sarcoidosis (SA) and idiopathic pulmonary fibrosis (IPF) is unclear. In the present study, we characterized the cytoskeletal proteins and the contraction properties in alveolar-associated fibroblasts recovered by bronchoalveolar lavage (BAL) in the two diseases. Afb were studied from BAL cells in eight IPF and seven SA patients. Cytoskeletal proteins were identified by ELISA and immunofluorescent methods. Biochemical measurements were done by dry chemistry. Contraction was performed by a gel contraction assay. Afb alpha-SM actin measured by ELISA was higher in IPF than in SA (p = 0.042). Vimentin, desmin, myosin, and fibroblast markers were expressed equally. Only in IPF did the Afb reveal the myofibroblast phenotype showing alpha-SM actin immunofluorescence labeling and, by electron microscopy, filaments with associated dense bodies with rough endoplasmic reticulum. Gel contraction showed that cells in IPF contracted significantly more than in SA (p = 0.046 IPF versus SA). The addition of ET-1 increased contraction in all groups. Dry chemistry analysis showed higher levels (p = 0.0065) of creatine phosphokinase (CPK), lower levels of glucose (p = 0.0082), and similar levels of Ca(2+) and lactate in the IPF and SA Afb. Dinitrofluorobenzene (DNFB), a potent inhibitor of CPK, completely abolished spontaneous cell contraction. Afb differentiates into myofibroblasts with different biochemical and energetic properties in IPF. Moreover, Afb from IPF patients showed increased contractile properties. This may explain the difference in the behavior patterns and outcomes of the two diseases. PMID- 11733854 TI - Increased sputum IL-8 and IL-5 in asymptomatic nonspecific airway hyperresponsiveness. AB - Since asymptomatic, nonspecific airway hyperresponsiveness (BHR) may be due to an enhanced local inflammatory response, we studied molecular markers of inflammation in induced sputum from subjects with asymptomatic BHR (n = 14) compared with control subjects (n = 13) and patients with chronic obstructive pulmonary disease (COPD) (n = 10). Pulmonary lung function parameters were measured by spirometry and body plethysmography. Hyperresponsiveness was defined based on histamine challenge. Induced sputum samples were collected and the solid phase was isolated and analyzed for leukocyte numbers and differential and for cytokines (ELISA). IL-8 was 2.4-fold increased (p = 0.036) in the sputum of subjects with asymptomatic BHR (24.8 +/- 22.0 ng/mL; +/- SD) and 11.2-fold enhanced in patients with COPD (117.8 +/- 106.3 ng/mL) as compared with control subjects (10.5 +/- 7.7 ng/mL). In control subjects, no IL-5 was measured, however, sputum of those with asymptomatic BHR contained IL-5 at 0.044 +/- 0.090 ng/mL fluid and COPD patients at 1.00 +/- 2.01 ng/mL. GM-CSF could not be detected in sputum samples of any subjects investigated. Number of total leukocytes was higher in those with asymptomatic BHR and COPD (with BHR: 9.4 +/- 10.8 x 10(5); COPD: 83.5 +/- 182.5 x 10(5)) compared with persons without BHR (2.9 +/- 3.4 x 10(5)). PMN were increased in patients with asymptomatic BHR (4.1 +/- 5.3 x 10(5)) (38.8 +/- 24.7%) and COPD (32.9 +/- 71.0 x 10(5)) (75.4 +/- 18.6%) compared with controls (0.7 +/- 0.9 x 10(5)) (25.8 +/- 25.7%). In contrast to PMN counts in those with asymptomatic BHR (0.06 +/- 0.11 x 10(5)) (1.5 +/- 3.7%), eosinophil counts were only slightly increased compared with control subjects (0.01 +/- 0.02 x 10(5)) (0.6 +/- 0.9%). This study supports the hypothesis that BHR in asymptomatic people is associated with airway inflammation that may predispose to development of chronic diseases such as COPD. PMID- 11733855 TI - Hygroma colli cysticum: prenatal diagnosis and prognosis. AB - The objective of this study is to analyze the prenatal management and prognostic factors of hygroma colli cysticum by using cytogenetic tests and sonographic morphological features. All cases with hygroma colli cysticum diagnosed and managed at our Prenatal Diagnostics Unit between January 96 and September 2000 were analyzed. Sonographic morphological features were divided in two groups; nonseptated (n = 18) and septated (n = 12) hygroma colli cysticum lesions were compared with fetal karyotype results and pregnancy outcome data. Statistical analysis was performed by Chi-square test and statistical significance was defined as p <0.05. In 5 years, 30 cases with hygroma colli cysticum were identified. Cytogenetic results were obtained from 23 (76.7%) cases (four chorionic villus sampling and 19 amniocentesis). Chromosomal abnormalities were present in 13 cases (56.5%). The most common chromosomal abnormality was Turner Syndrome (four cases, 17.4%) and Trisomy 18 (four cases, 17.4%). Pregnancy outcome data were available for 29 patients. Those fetuses with septated hygroma colli cysticum tended to have a worse fetal outcome, without statistical significance (p >0.05), compared with the nonseptated hygroma colli cysticum cases (75 vs. 61.1%, respectively). Fetal hygroma colli cysticum, either septated or not, carries high risks of aneuploidies and adverse fetal outcome. Recommended management includes karyotyping and if parents decide to continue the pregnancy ultrasound scan at 20 to 22 weeks' gestation is necessary, for excluding associated anomalies. At birth, if the cystic hygroma persists, it should be noted that a respiratory difficulty can happen and a pediatrician should standby as a precaution. PMID- 11733856 TI - Neonatal lupus erythematosus in association with anti-RNP antibody: a case report. AB - Neonatal lupus erythematosus (NLE) is a syndrome with the manifestation of dermatological, cardiac, hepatic, or hematological abnormalities. Thrombocytopenia has been documented infrequently in association with congenital heart block or lupus dermatitis in NLE. However, isolated neonatal thrombocytopenia may be the only manifestation of NLE. The strong association with maternal anti-SSA/Ro antibodies suggests a role for these antibodies in the pathogenesis of NLE. There are some data to suggest that anti-SSB/La and, rarely, anti-RNP antibodies play an important pathological role in some cases of NLE. The manifestation of anti-RNP-positive NLE was somewhat atypical. We report a case of anti-RNP-positive NLE with the manifestation of thrombocytopenia. Anti-SSA/Ro antibodies, which were negative based on the use of immunodiffusion, did exhibit low titer when later tested by enzyme-linked immunoadsorbent assay (ELISA). PMID- 11733857 TI - Outcome of twin pregnancies with extreme weight discordancy. AB - The objective of this paper is to examine the outcome of pregnancies with extreme weight-discordant twins. Percentage of birth weight discordancy was defined as the birth weight difference between the twins divided by the larger twin's weight and multiplied by 100. Discordancy was calculated for all twin births in which both fetuses were live born. In 33 pairs, the discordancy was defined as extreme (>35%) and they constituted the study group. Thirty-three pairs of twin defined with mild weight discordancy (15-25%), and 33 pairs defined as concordant to birth weight (<15% difference) were matched to the study group patients based on gestational age at delivery (+/- 7 days) and on the mode of delivery, and constituted the control groups. The records of all the patients were reviewed for pregnancy complications and for major and minor neonatal outcome variables. Significantly more parturients in the study groups were primiparous undergoing in vitro fertilization treatments to conceive. Significantly more women in the study group had severe preeclampsia compared with women with mild discordancy or concordant twins (12.1 vs. 3.0% and 0%, respectively, p <0.025). No significant differences were encountered between the groups in neonatal mortality or morbidity factors except an increased rate of hyperbilirubinemia in the study group, p = 0.006. Using logistic regression analysis, discordancy was not defined as an efficient predictor for adverse neonatal outcome. Twin pregnancies with extreme discordancy have a favorable neonatal outcome in correlation with gestational age and not with the percentage of discordancy. PMID- 11733858 TI - A reappraisal of neonatal blood chemistry reference ranges using the Nova M electrodes. AB - The objective of this study is to establish new reference ranges for whole blood electrolytes, gases, and selected chemistries in normal healthy newborn infants obtained from cord blood and at 2-4 hours of life based on modern analytic techniques. Healthy appropriate-for-gestational age (AGA), term infants were studied. Whole venous cord blood and blood drawn at 2-4 hours of life were analyzed for gases, sodium, potassium, chloride, ionized calcium, ionized magnesium, hemoglobin, and hematocrit. Concentrations for blood glucose and blood urea nitrogen were close to older published concentrations. Na(+), K(+), and Cl( ) were significantly different and had much narrower normal margins. New norms are included for blood gases, lactate, iCa, and iMg. We offer a range of values for often-measured blood chemistries in term, healthy infants, using modern laboratory methods. These values could serve as more up-to-date references compared with older values found in major textbooks of Neonatology. PMID- 11733859 TI - Lung agenesis in a neonate presenting with contralateral mediastinal shift. AB - A neonate with right lung agenesis presenting with respiratory distress is described. The unusual radiological features were contralateral mediastinal shift (in contrast to expected ipsilateral shift) and diaphragmatic eventration on the affected side. Mediastinal shift to the opposite side was due to intrathoracic hepatic herniation under a high placed eventrated diaphragm. Both these features have not been reported in association with lung agenesis to date. The authors have discussed other causes of respiratory distress in newborns that can cause mediastinal shift and have urged a high degree of clinical suspicion to pick up the cases with lung agenesis. The newer diagnostic modalities and the causes of mortality in neonates with this anomaly have also been highlighted. PMID- 11733860 TI - Prenatal diagnosis of midgut volvulus by sonography and magnetic resonance imaging. AB - We present a case of congenital midgut volvulus detected by prenatal sonography and ultrafast magnetic resonance (MR) imaging. At 34 weeks of gestation, enlarged hyperechogenic loops without peristalsis was identified by sonographic examination. On ultrafast T2-weighted single-shot fast-spin echo MR imaging, enlarged loops exhibited a lower signal intensity than the surrounding bowel loops, suggesting intraluminal hemorrhage. At explorative laparotomy following delivery, midgut volvulus causing hemorrhagic necrosis was found. Combined use of sonography and ultrafast MR imaging is useful to identify fetal midgut volvulus with hemorrhagic change. PMID- 11733861 TI - Relationship between fetal pulmonary maturity assessment and neonatal outcome in premature rupture of the membranes at 32-34 weeks' gestation. AB - The absence of fetal pulmonary maturity in patients with preterm premature rupture of the membranes (PPROM) is often considered an indication for conservative management. The purpose of this study was to examine the value of biochemical pulmonary maturity assessment for the prediction of neonatal outcome in patients with PPROM between 32 and 34 weeks' gestation. Pregnancies complicated by PPROM at 32 to 34 weeks' gestation that delivered from January 1995 to May 2000 and had biochemical pulmonary maturity assessment were reviewed. Patients with medical disorders, multiple gestations, fetal growth restriction or structural anomalies, or evidence of intra-amniotic infection were excluded. Neonatal outcome measures were compared between patients with mature and immature pulmonary indices. During this time period, 244 patients with PPROM at 32-34 weeks' gestation were delivered; 78 patients met inclusion criteria (n = 41 patients with mature indices and n = 37 patients with immature indices). There were no cases of perinatal death or sepsis. There was no difference in major neonatal morbidities including need for mechanical ventilation, grade 2 or 3 necrotizing enterocolitis, grade 3 or 4 intraventricular hemorrhage, or seizures. After controlling for confounding factors including gestational age at PPROM and delivery, latency period, group B streptococcus (GBS) vaginal colonization, corticosteroid therapy, neonatal sex, mode of delivery, fetal indications for delivery, and umbilical cord pH, biochemical pulmonary maturity was not predictive of major neonatal morbidity. In our population, biochemical pulmonary maturity status does not appear to be predictive of neonatal morbidity in pregnancies complicated by PPROM at 32-34 weeks' gestation. PMID- 11733862 TI - Unilateral diaphragmatic palsy in association with a subclavian vein thrombus in a very-low- birth-weight infant. AB - Phrenic nerve paralysis due to iatrogenic injury is not uncommon in neonates. We report an unusual case of acquired unilateral paralysis of the diaphragm in a very-low-birth-weight infant, associated with a thrombus secondary to percutaneous subclavian vein catheterization. Conservative management resulted in complete resolution of the paralyzed diaphragm. PMID- 11733863 TI - Can risk factor assessment replace universal screening for gonorrhea and Chlamydia in the third trimester? AB - The objective of this study is to determine whether risk factor (RF) screening can be used instead of routine third-trimester testing for gonorrhea and chlamydia in a clinic setting. We performed prospective analysis of women entering prenatal care over a 10-month period. Patients included received combined gonorrhea and chlamydia screening by DNA probe at the first prenatal visit and in the third trimester. RF examined included age <20, marital status, history of sexually transmitted disease (STD) or hepatitis, drug use and gestational age at entry into prenatal care. Only women with negative initial screens were included in univariate and multivariate analyses. Five hundred forty two women fulfilled study criteria. Sixty percent had 1 RF; 35% had a history of STD. Third-trimester screens were positive in 4.1%. The absence of RF had a negative predictive value of 99.1%. The sensitivity and specificity of RF screening was 90.9 and 40.7%, respectively (p = 0.003). The model of best fit was obtained using any positive RF and teen as independent variables (relative risk 6.9, 95% confidence interval 1.6-29.6, p = 0.01). In an urban clinic population, comprehensive RF screening is effective in predicting women at low risk for STD in the third trimester after an initial negative test. PMID- 11733864 TI - Role of three FKHR phosphorylation sites in insulin inhibition of FKHR action in hepatocytes. AB - Insulin inhibits insulin-like growth factor binding protein-1 (IGFBP-1) transcription by preventing FKHR protein family members from binding a specific insulin response element in the IGFBP-1 promoter. In most cells, three serine/threonine moieties in FKHR family members are phosphorylated after insulin treatment or protein kinase B/Akt (PKB) transfection, and each of the three phosphorylated PKB sites contributes to insulin- or PKB-mediated inhibition of both the action and the nuclear localization of FKHR family members. In hepatocytes, however, the middle PKB site (PKB2) of FKHR was required for insulin to phosphorylate FKHR and was the only PKB site that participated in insulin inhibition of FKHR action, indicating that insulin utilizes a unique pathway to regulate FKHR action in hepatocytes. In studies presented here, plasmids expressing native or mutant FKHR forms, either with or without N-terminal fusion to green fluorescent protein (GFP), were transiently transfected into HEP G2 cells. All FKHR forms stimulated IGFBP-1 promoter activity, and mutating any of the three FKHR PKB sites impaired the ability of insulin both to inhibit FKHR stimulated IGFBP-1 promoter activity and to induce FKHR accumulation in cytoplasm. Thus, in hepatocytes as in other cell lines, all three FKHR PKB sites participate in insulin-mediated inhibition of FKHR action and in insulin-mediated accumulation of FKHR in cytoplasm. PMID- 11733865 TI - Interleukin-1 beta inhibits proinsulin conversion in rat beta-cells via a nitric oxide-dependent pathway. AB - Exposure of pancreatic beta-cells to interleukin-1 beta (IL-1 beta) alters their protein expression and phenotype. Previous work has shown that IL-1 beta inhibited proinsulin conversion in rat islets, but the mechanism of this inhibition remained unknown. To investigate this phenomenon further, we examined purified rat beta-cells for IL-1 beta-induced inhibition of proinsulin conversion and nitric oxide (NO)-dependency of this inhibitory process. Rat beta-cells were cultured for 24 h with or without IL-1 beta and the inducible-nitric-oxide synthase (iNOS) inhibitor N(G)-methyl-L-arginine (NMA). Exposure to IL-1 beta suppressed proinsulin-1 and proinsulin-2 synthesis by more than 50 %. Conversion of both proinsulin isoforms was also delayed. The suppressive effects of IL-1 beta on proinsulin synthesis and conversion were prevented by addition of NMA. Exposure to IL-1 beta also decreased the expression of the proinsulin convertase (PC) PC2. This decrease in PC2 expression was NO-dependent. In conclusion, IL-1 beta inhibition of proinsulin conversion in rat beta-cells occurs via an NO mediated pathway. PMID- 11733866 TI - Gender differences in the vasomotor effects of different steroid hormones in rat pulmonary and coronary arteries. AB - It is well recognised that oestrogens possess vasodilatory properties, and similar responses to testosterone have been demonstrated. However, vasomotor effects of other steroid hormones have not been well described. Direct comparisons of the relative vasoactivity of different steroid hormones in different vascular beds in male and female genders have not been made. Coronary and pulmonary arteries from adult Wistar rats were mounted in a wire myograph, loaded to 100 and 17 mmHg respectively, maximally pre-contracted with 1 x 10(-4) M prostaglandin-F-2-alpha, and dose response curves to 1 x 10(-6) to 1 x 10(-3) or 3 x 10(-3) M of 17 beta-oestradiol, testosterone, progesterone, and cortisol dissolved in water were constructed. Addition of each steroid hormone caused acute, dose dependent dilatation in coronary and pulmonary vessels. In coronary arteries the order of activity was testosterone > progesterone > 17 beta oestradiol > cortisol, p < 0.001. In pulmonary arteries, the order of activity was progesterone > testosterone > cortisol > 17 beta-oestradiol, p < 0.001. Pulmonary arteries from male animals were more sensitive to the effects of testosterone than those from female animals, p = 0.003, whereas coronary arteries from female animals were more sensitive to the effects of 17 beta-oestradiol than those from male animals, p < 0.001. We have demonstrated significant differences in the in vitro vasomotor effects of different steroid hormones in two distinct vascular beds. Gender differences in vasomotor responses to steroid hormones may play a role in the aetiology of vasospastic diseases. PMID- 11733867 TI - Enzyme immunoassay of 17 beta-estradiol, estrone conjugates, and testosterone in urinary and fecal samples from male and female mice. AB - ELISA measures of 17 beta-estradiol, estrone conjugates, and testosterone were adapted for fecal and urinary samples from laboratory mice. We will report on validations of these assays and data from interacting males and females. Unconjugated gonadal steroids were consistently measurable in urine and feces of both males and females. Females that were parturient following insemination excreted relatively low levels of urinary testosterone compared to non-parturient females. The results are consistent with evidence that elevated androgens and estrogens are incompatible with intrauterine implantation of fertilized ova, and suggest that steroids in male urine could contribute to pheromonal action. These methods permit repeated noninvasive measurement of steroid activity in this species. PMID- 11733868 TI - Indomethacin does not affect endogenous glucose production in type 2 diabetes mellitus. AB - In healthy subjects, basal endogenous glucose production is partly regulated by paracrine intrahepatic factors. It is currently unknown whether paracrine intrahepatic factors also influence the increased basal endogenous glucose production in patients with type 2 diabetes mellitus. Administration of indomethacin to patients with type 2 diabetes mellitus stimulates endogenous glucose production and inhibits insulin secretion. Our aim was to evaluate whether this stimulatory effect on glucose production is solely attributable to inhibition of insulin secretion. In order to do this, we administered indomethacin to 5 patients with type 2 diabetes during continuous infusion of somatostatin to block endogenous insulin and glucagon secretion and infusion of basal concentrations of insulin and glucagon in a placebo-controlled study. Endogenous glucose production was measured 3 hours after the start of the somatostatin, insulin and glucagon infusion, for 4 hours after administration of placebo/indomethacin, by primed, continuous infusion of [6,6-(2)H(2)] glucose. At the time of administration of placebo or indomethacin, there were no significant differences in plasma glucose concentrations and endogenous glucose production rates between the two experiments (16.4 +/- 2.09 mmol/l vs. 16.6 +/- 1.34 mmol/l and 17.7 +/- 1.05 micromol/kg/min and 17.0 +/- 1.06 micromol/kg/min), control vs. indomethacin). Plasma glucose concentration did not change significantly in the four hours after indomethacin or placebo administration. Endogenous glucose production in both experiments was similar after both placebo and indomethacin. Mean plasma C-peptide concentrations were all below the detection limit of the assay, reflecting adequate suppression of endogenous insulin secretion by somatostatin. There were no differences in plasma concentrations of insulin (76 +/- 5 vs. 74 +/- 4 pmol/l) and glucagon (69 +/- 8 vs. 71 +/- 6 ng/l) between the studies with levels remaining unchanged in both experiments. Plasma concentrations of cortisol, epinephrine, and norepinephrine were similar in the two studies and did not change significantly. We conclude that indomethacin stimulates endogenous glucose production in patients with type 2 diabetes mellitus by inhibition of insulin secretion. PMID- 11733869 TI - Increased renal GLUT1 abundance and urinary TGF-beta 1 in streptozotocin-induced diabetic rats: implications for the development of nephropathy complicating diabetes. AB - Increased expression of transforming growth factor beta-1 (TGF-beta 1) and glucose transporter (GLUT1) has been implicated in the genesis of diabetic nephropathy. The aim of this study was to evaluate GLUT1 protein levels in the renal cortex of a rat model of diabetes as well as its relationship to urinary albumin and TGF-beta1. Streptozotocin-injected rats (n = 13) and controls (n = 13) were compared for their urinary albumin, and TGF-beta 1 and for renal cortical and medullar GLUT1 protein abundance. GLUT1 protein content was determined by optical densitometry after Western blotting using an anti-GLUT1 antibody; urinary albumin was measured using electroimmunoassay, urinary TGF-beta 1 using ELISA. Forty-five days of diabetes resulted in increased albuminuria (p < 0.05), urinary TGF-beta 1 (p < 0.05) and GLUT1 protein abundance (p < 0.05). There was a positive correlation between urinary TGF-beta 1 and plasma glucose levels (r = 0.65, p < 0.05) and albuminuria (r = 0.72, p < 0.05). We concluded that 45 days of diabetes result in incipient diabetic nephropathy and increased cortical GLUT1 protein abundance. We speculate that the higher cortical GLUT1 protein levels in diabetes may amplify the effects of hyperglycemia in determining higher intracellular glucose in mesangial cells, thereby contributing to diabetes-related kidney damage. PMID- 11733870 TI - Body composition assessment in type 1 diabetes mellitus patients over 15 years old. AB - BACKGROUND: The aim of the present study was to assess the anthropometric characteristics and body composition in type 1 diabetic patients and compare the results with a randomly selected control population. MATERIAL AND METHODS: We studied 75 type 1 diabetic patients, 43 male and 32 female, recruited from consecutive diagnosed type 1 diabetic patients attending the Endocrine Unit and treated with a intensive insulin regimen, and 93 control subjects, 44 males and 49 females representative of the census of this city. We performed a dietary recall in patients and determined anthropometric characteristics, both in patients and controls, body weight, height, body-mass index, waist-hip ratio and body composition parameters: total body water, free-fat mass, body free-fat mass, fat mass and body fat by bioelectrical impedance analyser. RESULTS: In diabetic male patients, we observed lower waist-hip ratio than in controls, 0.84 +/- 0.06 vs. 0.88 +/- 0.07, p = 0.021, higher free-fat mass in female diabetic patients, 48.5 +/- 5.6 vs. 45.6 +/- 5.9 kg, p = 0.03, lower fat mass in male diabetic patients, 9.5 +/- 6.9 vs. 14.6 +/- 8.5 kg, p = 0.003. We did not find any correlation among the parameters of body composition and dietary macronutrient intake in patients. CONCLUSIONS: The present study exposes the differences in anthropometric characteristics and body composition in type 1 diabetes mellitus, especially lower waist-hip ratio in male, higher free-fat mass in female and lower fat mass in male. PMID- 11733871 TI - Effect of insulin on von Willebrand factor release in normal and diabetic subjects: in vivo and in vitro studies. AB - The aim of this study was to evaluate the effect of insulin on the release of vWf in vivo during an oral glucose tolerance test (OGTT) performed in normal, glucose intolerant and diabetic subjects and in vitro on human endothelial cells. Twenty eight subjects exhibiting risk factors for diabetes underwent an OGTT: 11 subjects proved to be normal, 7 were glucose-intolerant and 10 diabetic. In each group, the vWf and PAI-1 plasmatic levels were measured at t = 0, 30 min and 180 min after the beginning of the test. Human endothelial cells from non-diabetic and diabetic subjects were incubated in the presence of human insulin at various concentrations (0.25, 2.5, 25 and 250 mUI/ml). After 1, 4, and 24 hours of incubation, the release of vWf and endothelin 1 was measured in the cell supernatant and the intracellular amount of vWf in the cell lysate. During the OGTT, the vWf levels in plasma were not affected despite a 4.5-, 6-, and 2.5-fold increase in insulin levels in normal, glucose-intolerant and diabetic subjects, respectively. Although raised in all three groups, PAI-1 plasmatic levels remained constant during the test. After 24 hours of exposure to insulin (0.25 mU/ml), the release of vWf by endothelial cells reached 35.94 +/- 23.08 % of the basal value for non-diabetic subjects, and 27.57 +/- 10.05 % for diabetic patients. Similar results were observed in non-stimulated cells. Insulin had no influence on intracellular vWf content, which remained comparable to control values. Regardless of its concentration, insulin failed to stimulate the release of vWf by endothelial cells of non-diabetic and diabetic subjects, while its ability to stimulate the release of endothelin 1 was preserved. In conclusion, hyperinsulinemia had no adverse effect on circulating vWf in normal or diabetic subjects. Neither release nor intracellular vWf content in non-diabetic or diabetic endothelial cells was influenced by insulin in vitro. PMID- 11733872 TI - Novel male mice show gradual decline in the capacity to disrupt early pregnancy and in urinary excretion of testosterone and 17 beta-estradiol during the weeks immediately following castration. AB - Novel male mice can disrupt intrauterine implantation of fertilized ova in inseminated females. Evidence indicates mediation by androgen-dependent excretions. This study was designed to examine the time course of males' ability to disrupt pregnancy following castration and relate this to their urinary excretion of testosterone and 17 beta-estradiol. During days 1-5 of pregnancy, previously inseminated females were housed underneath castrated novel males at various intervals after the surgery. Castrated males generally continued to disrupt pregnancy during the initial weeks after surgery. Progressively, the probability of retention of pregnancy increased as a linear function of time since castration. There was an apparent asymptote, where the majority of females remained pregnant, beginning at about 6 weeks following surgery. Males' excretion of testosterone and 17 beta-estradiol in urine, measured via ELISA procedures, diminished gradually during the weeks after castration. PMID- 11733873 TI - Long-term correction of hyperglycaemia in streptozotocin-induced diabetic rats transplanted with islets placed in an implantation device. PMID- 11733874 TI - Octreotide inhibits the secretion of interleukin-12 from mononuclear cells in human peripheral blood (PBMCs) in vitro. PMID- 11733875 TI - Measurement of upper extremity volume in women after axillary dissection for breast cancer. AB - OBJECTIVE: To determine if 2 methods of calculating upper extremity volume (using arm circumferences) can substitute for water displacement volumetry. DESIGN: Interrater and test-retest reliability and limits of agreement for volume measures. SETTING: University. PARTICIPANTS: Twenty-five women at risk for lymphedema who had undergone axillary lymph node dissection surgery for breast cancer. INTERVENTIONS: Circumference and volume measurements of both upper extremities were taken by 2 physical therapists at an initial visit and by 1 of the therapists 1 week later. MAIN OUTCOME MEASURES: Intraclass correlation coefficients (ICCs) were calculated to analyze measurement reliability. Pearson's product-moment correlation coefficient (r) was used to evaluate the relationship between volumetry and calculated truncated cone volumes. Limits of agreement were calculated to determine the level of agreement between the 2 measurement methods. RESULTS: Interrater and test-retest reliability ICCs for circumferential and volumetric data were .99 and .99, respectively. Pearson's r values were .93 and .97 for the single truncated cone and the summed truncated cone volume calculations, respectively. Limits of agreement (mean +/- 2 standard deviations) were -52 +/- 334mL and -40 +/- 234mL, respectively, between volumetry and the single truncated cone and summed truncated cone calculations. CONCLUSIONS: Calculated and volumetric measurements in this population are both reliable and closely related, but do not agree with each another, and thus should not be used interchangeably. PMID- 11733876 TI - Depressive symptoms and history of depression predict rehabilitation efficiency in stroke patients. AB - OBJECTIVE: To examine how depressive symptoms, a history of depression, and cognitive functioning contribute to the prediction of rehabilitation efficiency in stroke patients. DESIGN: Consecutive admissions to an acute inpatient rehabilitation program were screened for cognitive functioning and level of depressive symptoms. History of depression was determined by family member interview. Functional status was evaluated at time of admission and discharge. Depressive symptoms, depression history, and cognitive functioning were examined as predictors of length of stay (LOS) and efficiency of utilization of rehabilitation services. SETTING: Acute inpatient rehabilitation hospital. PATIENTS: A total of 348 consecutive stroke admissions to an inpatient program were evaluated for depression and cognitive functioning, of whom 243 patients completed all aspects of the screening. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Rehabilitation progress, measured with the LOS efficiency measure (LOS-EFF) of the FIM instrument, and length of rehabilitation hospital stay. RESULTS: Patients with higher levels of depressive symptoms used rehabilitation services less efficiently than those with lower symptom levels but did not have longer LOSs. History of depression was associated with longer LOS and less efficient use of rehabilitation services. Cognitive impairment did not predict rehabilitation efficiency. CONCLUSIONS: The present study lends further support to the benefits of screening stroke patients at the time of rehabilitation admission for depression and history of depression. Identifying patients who have high levels of depressive symptoms and/or a previous depressive episode will allow more comprehensive assessment and rapid intervention. PMID- 11733877 TI - Symmetrical body-weight distribution training in stroke patients and its effect on fall prevention. AB - OBJECTIVE: To determine the role of symmetrical body-weight distribution training in preventing falls among patients with hemiplegic stroke. DESIGN: A prospective study using a standing biofeedback trainer. SETTING: Hospital-based rehabilitation units. PATIENTS: Fifty-four patients with hemiplegic stroke (30 in the training group, 24 in the control group). INTERVENTIONS: Conventional stroke rehabilitation program, plus symmetrical standing training and repetitive sit-to stand training, with a standing biofeedback trainer. Training effect was evaluated by assessing the sit-to-stand performance and comparing the occurrence of falls in the 2 groups at a 6-month follow-up. MAIN OUTCOME MEASURES: Occurrence of falls, sit-to-stand performance, including body-weight distribution, rate of rise in force, and sway in center of pressure (COP). RESULTS: Significant improvement in sit-to-stand performance was found in patients in the training group. Body weight was distributed more symmetrically in both legs, with less mediolateral sway in the COP when rising and sitting down. The mean difference in body-weight distribution between the left and right legs while subjects were rising from a chair significantly decreased, from 49.5% +/- 18.9% to 38.6% +/- 15.8% of body weight (BW) (p < .005). The rate of rise in force while rising from a chair significantly increased, from 28.3% +/- 13.5%BW/s to 53.6% +/- 20.5%BW/s (p < .001). At the 6-month follow-up, 10 of 24 patients (41.7%) in the control group had fallen, compared with only 5 of 30 patients (16.7%) in the training group (p < .05). CONCLUSIONS: Symmetrical body-weight distribution training may improve sit-to-stand performance and, consequently, decrease the number of falls by stroke patients. PMID- 11733878 TI - Head and pelvic movements during a dynamic reaching task in sitting: implications for physical therapists. AB - OBJECTIVES: To describe the distance reached, speed, and movement of the head and pelvis of healthy volunteers; to describe any influence of age on these variables; and to compare healthy volunteers and subjects with hemiplegia while performing a seated reaching task. DESIGN: Age-matched, case-control study. SETTING: Gait laboratory in a general hospital. PARTICIPANTS: A convenience sample of 53 healthy volunteers (30 women; 23 men; mean age, 57yr; range, 30 79yr) and 5 subjects with hemiplegia (2 women, 3 men; mean age, 65yr; range, 60 78yr) were recruited within 6 weeks poststroke. INTERVENTIONS: Participants sat on a bench with feet supported and reached laterally as far as they could without falling. MAIN OUTCOME MEASURES: The speed, distance reached, and angular movements of the head and pelvis were recorded by using the 3-dimensional movement analysis system. RESULTS: A significant age-related reduction in the distance reached (p < .001), velocity of the movement (p =.000), and pelvic tilt used (p < .01) was found among healthy volunteers. Comparison of data from healthy volunteers and subjects with hemiplegia showed a significant reduction in the angular movements of the heads of subjects with hemiplegia. CONCLUSIONS: The findings suggest conservation of movement with increasing age and stroke. This movement reduction could have negative effects on a subject's ability to make postural changes in response to disturbance and activity. Such information may assist therapists to gain insight into the nature of balance deficits and the adaptive behavior that could result. PMID- 11733879 TI - Comparison of effortful and noneffortful swallows in healthy middle-aged and older adults. AB - OBJECTIVE: To assess the effects of effortful swallowing, a common compensatory strategy for dysphagia, on the bolus and swallowing mechanism of middle-aged and older men and women. DESIGN: Case-controlled design in which subjects completed both the intervention technique and the control behavior. SETTING: A university hospital. PARTICIPANTS: Sixty-four healthy men and women between 45 and 93 years of age from the community. INTERVENTIONS: Participants swallowed 3-mL thin liquid boluses both normally and using the effortful swallow strategy. MAIN OUTCOMES MEASURES: The biomechanics and bolus flow patterns of swallows were analyzed from videofluoroscopic and simultaneous oral pressure data. RESULTS: Subjects at all ages generated significantly increased oral pressures at each sensor location using the effortful swallow (p = .0001), with the pressure increase greater for the middle-aged subjects compared with older subjects. Several durational measures were significantly longer with the effortful swallow including: hyoid maximum anterior excursion (p < .04), laryngeal vestibule closure (p < .0001), and duration of the upper esophageal sphincter opening (p =.0001). The hyoid bone moved further in the superior direction with the effortful swallow (p = .002). There was a trend of decreased oral residue with the effortful swallow (p = .06). CONCLUSION: Biomechanical and bolus flow aspects of swallowing changed when healthy individuals performed effortful swallows with 3-mL boluses. PMID- 11733880 TI - Relationship between knee extension force and stand-up performance in community dwelling elderly women. AB - OBJECTIVE: To describe the relationship between knee extension force and stand-up performance. DESIGN: Prospective, cross-sectional, descriptive, and correlational. SETTING: Community. PARTICIPANTS: Fifty-five community-dwelling women 60 or more years old. INTERVENTIONS: Information pertaining to stand-up performance was gathered via structured questions. The time to complete 1 sit-to stand from a bench of known height (16in) was obtained in conjunction with subjects' ratings of perceived exertion (RPEs) relative to the task. Bilateral lower extremity knee extension strength was measured with a hand-held dynamometer and normalized against body weight. MAIN OUTCOME MEASURES: Total knee extension forces (raw and body-weight normalized values) generated by each subject were correlated with 3 measures of stand-up performance. RESULTS: Low to moderate correlations (r = -.323 to -.526) were found between knee extension forces and the stand-up performance measures of bench stand time and stand-up difficulties. Normalized knee extension force had a higher correlation than raw knee extension force with stand-up difficulties and RPE. Correlations were higher when the curvilinear nature of the relationship was taken into account. The highest correlation was between normalized knee extension force and the RPE associated with the bench stand task (r = -.767). CONCLUSION: Knee extension force offers an important but incomplete explanation of stand-up performance from low surfaces by elderly women. Interventions leading to increased knee extension force may improve stand-up performance. PMID- 11733881 TI - Vertical impact increase in middle age may explain idiopathic weight-bearing joint osteoarthritis. AB - OBJECTIVE: To test the hypothesis that ground reaction force increases when a standard stepping task is performed in late middle age. DESIGN: Consecutive sample. SETTING: Internal medicine practice. PARTICIPANTS: Thirty-six male patients (age range, 17-72yr) from an internal medicine practice. INTERVENTION: Subjects performed 20 consecutive footfall impacts onto a force platform while barefoot and while wearing shoes. MAIN OUTCOME MEASURES: Ground reaction forces were recorded for each footfall. RESULTS: Impact is positively related to age both when barefoot (r = .84, p < .001) and when wearing shoes (r = .71, p < .001). Impact remains constant until age 50, after which it increases by 13.3% for barefoot subjects. Barefoot impact was significantly lower and less variable than impact when shod (barefoot = 1.18 body weight [BW]; shod = 1.22 BW; F(1,5) = 169.91, p < .001). CONCLUSION: An increase in impact force during locomotion was identified that occurs in late middle age, when stability declines and idiopathic weight-bearing joint osteoarthritis develops. Because impact is negatively related to stability, the impact rise is probably caused by postural adjustments to instability resulting from irreversible neurologic decline. This heightened impact may account for the accelerated rate of weight-bearing joint osteoarthritis that begins in late-middle age. PMID- 11733882 TI - Submaximal exercise capacity and maximal power output in polio subjects. AB - OBJECTIVES: To compare the submaximal exercise capacity of polio subjects with postpoliomyelitis syndrome (PPS) and without (non-PPS) with that of healthy control subjects, to investigate the relationship of this capacity with maximal short-term power and quadriceps strength, and to evaluate movement economy. DESIGN: Cross-sectional survey. SETTING: University hospital. PARTICIPANTS: Forty three polio subjects (25 PPS, 18 non-PPS) and 12 control subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Power output, oxygen uptake, and heart rate were measured in an incremental submaximal cycle ergometry test. Maximal short-term power was measured in 5-second all-out efforts. Knee extensor strength was measured on a chair dynamometer. RESULTS: The mean submaximal power +/- standard deviation at 80% of heart rate reserve of 83.8 +/- 29.9 watts in the polio subjects was significantly less than the mean submaximal power of 142.1 +/- 30.4 watts in the control group. However, expressed as a percentage of the maximal short-term power, submaximal power did not differ between the groups. Strength and maximal short-term power correlated significantly (p < .005) with submaximal power (r = .64 and .76, respectively). The oxygen uptake was higher than theoretically expected for the given submaximal power output in polio subjects, and appeared to increase with increasing asymmetry in strength and power between legs. No differences were found between PPS and non-PPS subjects. CONCLUSION: The submaximal work capacity of polio subjects was severely reduced, mainly in association with the reduced muscle capacity. And, because of a reduced movement economy, their energy cost was elevated. Although muscle loads in activities such as walking and climbing stairs differ from cycling, they also may require elevated relative levels of effort, predisposing subjects to premature fatigue in sustained activity. PMID- 11733883 TI - Arterial blood gases during exercise: validity of transcutaneous measurements. AB - OBJECTIVE: To investigate the validity of transcutaneous measurements of blood gas tensions for the assessment of partial arterial pressure of oxygen (PaO(2)) and carbon dioxide (PaCO(2)) during treadmill exercise. DESIGN: Experimental, self-controlled against a reference standard. SETTING: Lung function laboratory. PATIENTS: Eighty-one patients with various lung diseases. INTERVENTIONS: At rest and at symptom-limited peak exercise, puncture of the radial artery with concurrent transcutaneous measures of blood gases. MAIN OUTCOME MEASURES: Arterial blood samples were analyzed with a radiometer to measure PaO(2) and PaCO(2). A microgas apparatus was used to measure gas tensions transcutaneously. Values obtained transcutaneously (TcPO(2), TcPCO(2)) were compared with those obtained by blood sample. TcPO(2) was adjusted as close as possible to the PaO(2) obtained in the same conditions, with the correction factor of the apparatus. Values obtained transcutaneously were compared with those obtained by blood sample to establish the sensitivity and specificity of the noninvasive method. RESULTS: Mean differences +/- standard deviation between transcutaneous and arterial tension at peak exercise were 0.4 +/- 7.0mmHg and 2.1 +/- 3.3mmHg for PaO(2) and PaCO(2), respectively. The transcutaneous device enabled us to predict a decrease in PaO(2) (>or=2mmHg) from rest to exercise with a sensitivity of 92.1% and a specificity of 90% and an increase in PaCO(2) with a sensitivity of 88% and a specificity of 58.9%. CONCLUSIONS: Although transcutaneous measurement are sufficiently sensitive and specific to detect patients whose PaO(2) decreases during exercise, its precision is not sufficient for gas exchange calculations. PMID- 11733884 TI - Electromyographic biofeedback-controlled exercise versus conservative care for patellofemoral pain syndrome. AB - OBJECTIVE: To determine the effects of electromyographic biofeedback treatment in patients with patellofemoral pain syndrome. DESIGN: Randomized controlled trial. SETTING: A physical medicine and rehabilitation department in a research hospital of a university referral center. PATIENTS: Sixty patients with patellofemoral pain syndrome. Patients were randomly placed into 2 groups: biofeedback group (n = 30) and a control group (n = 30). INTERVENTION: The biofeedback group received electromyographic biofeedback training and a conventional exercise program, whereas the control group received a conventional exercise program only. MAIN OUTCOME MEASURES: Maximum and mean contraction values of the vastus medialis and the vastus lateralis muscles were assessed with the biofeedback device. Pain and functional status of the patients were measured by a visual analog scale (VAS) and the Functional Index Questionnaire (FIQ), respectively. RESULTS: Contraction values improved significantly at the end of the first month, compared with the pretreatment values in both groups. Mean contraction values in the biofeedback group of the vastus medialis muscles in all 3 monthly measurements, and the vastus lateralis muscles at the end of the first month, were significantly higher than those of the control group. Significant improvements were shown for both the VAS and the FIQ in both groups. Monthly follow-ups showed no VAS and FIQ differences between the groups. CONCLUSION: Electromyographic biofeedback treatment did not result in further clinical improvement when compared with a conventional exercise program in patients with patellofemoral pain syndrome. PMID- 11733885 TI - Locomotor-specific measure of spasticity of plantarflexor muscles after stroke. AB - OBJECTIVES: To study the stretch reflex excitability (spasticity) of the plantarflexor muscles during gait in patients with hemiparesis and to study the relationships of spasticity during gait with spasticity at rest and gait speed. DESIGN: Cross-sectional, descriptive. SETTING: Rehabilitation center. PARTICIPANTS: Convenience sample of 30 patients (58 +/- 11yr) with hemiparesis (<6mo poststroke) and 15 healthy controls (59 +/- 8yr). INTERVENTIONS: Patients walked at natural speed, healthy subjects at very slow speed for 10 gait cycles. Electromyographic activation of the medial gastrocremius was recorded by using surface electrodes. A 2-dimensional video camera system with reflective markers was used to acquire kinematics of the lower limbs. MAIN OUTCOME MEASURES: Electromyography-lengthening velocity slopes, calculated from measures obtained during the lengthening periods of the medial gastrocnemius muscle during the stance and the swing phases. Measured spatisticity (Modified Ashworth Scale [MAS]), static strength (ankle clonus), and motor control (Fugl-Meyer test). RESULTS: Velocity-sensitive electromyographic responses, indicative of hyperactive stretch reflexes, were found on the paretic side during the stance phase of gait (in 66% of the patients), but not on the nonparetic side or in controls. In many patients, velocity-sensitive responses coexisted with low plantarflexor activation levels during the stance phase. No clear patterns of response were measured during the swing phase in either group. Spasticity during gait in the patients was found to be positively related (r = .47, p < .01; r = .57, p < .001) to spasticity at rest (MAS; ankle clonus), whereas it was found to be negatively related to gait speed (r = -.47 to -.53, p < .01). CONCLUSIONS: The validity of the present method is supported by the fact that it is locomotor specific and that it allowed for a good discrimination between spastic and nonspastic limbs, as well as between stance and swing phases of the gait cycle. The results also support plantarflexor spasticity as a factor contributing to the poor locomotor performance after stroke. PMID- 11733886 TI - Pelvic and lower limb compensatory actions of subjects in an early stage of hip osteoarthritis. AB - OBJECTIVE: To determine if compensatory actions take place at the pelvis and other joints of the affected lower limb in subjects who were in an early stage of hip osteoarthritis (OA). DESIGN: Nonrandomized, case-control study. SETTING: A gait laboratory. PARTICIPANTS: Seventeen patients with OA of the hip (clinical group) matched with 17 healthy elderly subjects (nonclinical group). INTERVENTIONS: Video data obtained while subjects walked a 10-meter walkway twice and stepped across a forceplate. MAIN OUTCOME MEASURES: Four phasic and temporal gait parameters (walking speed, stance phase relative duration, stride length, cadence) 10 pelvic (pelvic tilt, obliquity, rotation at push-off maximum range of motion for all 3) and hip (3 hip angles at push-off, maximum hip flexion) kinematic parameters, 3 hip moments, and twenty-seven 3-dimensional peak muscle powers (labeled by joint, peak power, plane) developed in the lower limb joints during the gait cycle. RESULTS: Subjects in the clinical group were characterized by a 12.4% slower walking speed. The pelvis was more upwardly tilted (2.5 times) at push-off in the clinical group than in the nonclinical group. Obliquity, measured in the frontal plane, revealed that the pelvis dropped more (2.4 times) on the unsupported limb of the clinical group at push-off. In the sagittal plane, subjects in the clinical group absorbed less energy in their second hip peak power for decelerating the thigh extension and generated less hip pull (third hip peak power) than the nonclinical group by 34% and 29%, respectively. In the sagittal plane, the clinical group had 57% lower second knee peak power to straighten the joint shortly after heel strike, and 43% less knee absorption (third peak power) at push-off. During the push-off phase, the clinical group developed more than twice their third peak knee power in the frontal plane and 5 times more their third peak knee power in the transversal plane than the peak knee power of the nonclinical group in an attempt to control knee adduction and to facilitate body-weight transfer by an internal rotation. At the end of the swing phase, the fourth peak power in the sagittal plane showed the absorption power required to decelerate the leg; it was reduced by 35% in the clinical group, representing a strategy to increase walking speed by lengthening the stride length. CONCLUSIONS: Even at an early stage of hip OA, joint degeneration was compensated by an increase in pelvis motion and muscle power generation or absorption modifications in other lower limb joints. PMID- 11733887 TI - Effect of stroke on step characteristics of obstacle crossing. AB - OBJECTIVE: To compare spatial and temporal measures during lead limb obstacle crossing between subjects with stroke and healthy subjects. DESIGN: Experimental, observational, with matched controls. SETTING: Geriatric rehabilitation unit in a tertiary referral hospital. PARTICIPANTS: Distance data were available for 19 subjects with stroke and 19 able-bodied subjects. Temporal data were available for 16 subjects with stroke and 16 able-bodied subjects. Subjects with stroke were inpatients and had to be able to walk 10 meters without assistance or gait aid. INTERVENTION: Subjects were required to step over high and wide obstacles, ranging from 1 to 8cm, and trials were videotaped. MAIN OUTCOME MEASURES: Toe clearance, preobstacle distance, postobstacle distance, step length, proportion of step length preobstacle, step time, preobstacle step time, postobstacle step time, and proportion of step time preobstacle were measured. RESULTS: Mann Whitney U tests were performed to determine differences between the 2 groups. Subjects with stroke had significantly higher toe clearance, smaller postobstacle distances, and greater step times than healthy subjects. Subjects with stroke did not demonstrate a significant reduction in preobstacle distance. CONCLUSION: By modifying their lead limb trajectory during obstacle crossing, persons with stroke reduce the risk of a trip due to toe contact, but the modification may expose them to other safety risks. PMID- 11733888 TI - Societal risk factors associated with spinal cord injury secondary to gunshot wound. AB - OBJECTIVES: To determine (1) how many individuals with spinal cord injury (SCI) secondary to gunshot wound (GSW) had previous violent injury requiring medical professional intervention or were previously involved in the criminal justice system, and (2) how many had social service intervention initiated before their SCI to deter future injury. DESIGN: Follow-up survey of patients admitted to an inpatient SCI rehabilitation unit between 1990 and 1998. SETTING: An urban rehabilitation hospital. PARTICIPANTS: Fifty-six patients with SCI secondary to GSW. INTERVENTION: Survey questions inquired about previous GSW, violent injury, and criminal justice involvement. Subjects were queried about social service intervention. MAIN OUTCOME MEASURES: Number and percentage of respondents. RESULTS: Of the 56 respondents, 17 (30%) had a previous GSW; 9 (16%) had another violent injury requiring treatment in the emergency department; and 29 (52%) had prior involvement in the criminal justice system. A total of 55 opportunities for social service intervention were identified. Social service intervention was initiated only once. CONCLUSIONS: Most respondents had preinjury involvement in the criminal justice system and/or previous violent injury. In all of these situations except 1, there was failure to initiate social service intervention. These preliminary data suggest that criminal justice system involvement and violent injury are common among patients who have SCI secondary to GSW. More aggressive efforts at social service intervention could be an effective means to deter future disability. PMID- 11733889 TI - A decision pathway for the staged management of foot problems in diabetes mellitus. AB - This review describes a decision pathway for the conservative management of foot problems in persons with diabetes mellitus. The decision pathway was developed by the Louisiana State University Health Sciences Center Diabetes Foot Program as part of the Statewide Diabetes Disease Management Initiative to standardize foot care in a medically underserved population. The pathway describes the prevention and/or management of foot problems through 5 clinical subpathways: injury prevention, warm swollen foot, ulcer, osteomyelitis, and remodeling. The pathway provides physiatrists and other rehabilitation practitioners with a framework to evaluate and manage neuropathic foot problems in a multidisciplinary setting. PMID- 11733890 TI - Delayed diagnosis of concomitant rotator cuff tear and brachial plexopathy in a patient with traumatic brain injury: a case report. AB - Traumatic brain injury (TBI) is often accompanied by additional trauma that can be obscured by cognitive dysfunction or multiple injuries in the same region of the body. This report describes the case of an unhelmeted motorcycle rider who collided with a telephone pole. He sustained a diffuse subarachnoid hemorrhage, bilateral subdural hematomas (right frontal and left temporal), diffuse axonal injury in the subcortical and periventricular white matter, and a left tibial fracture. After medical and surgical stabilization, he was transferred to a subacute rehabilitation facility and then to a rehabilitation center. He was evaluated for pain and limited range of motion in his right shoulder, where both a rotator cuff tear and a brachial plexopathy were diagnosed. This report discusses concomitant injuries that occur with TBI, and the management of rotator cuff tears and brachial plexopathy. PMID- 11733891 TI - Combined functional magnetic resonance imaging and transcranial magnetic stimulation evidence of ipsilateral motor pathway with congenital brain disorder: a case report. AB - We present the case of 28-year-old man with schizencephaly who had mild left hemiparesis with mirror movement. Brain mapping using functional magnetic resonance imaging (fMRI) and transcranial magnetic stimulation (TMS) for both hand muscles was done to evaluate his neurologic state. Motor evoked potential (MEP) from both abductor pollicis brevis (APB) muscles was obtained simultaneously. fMRI showed that the left primary sensorimotor cortex became active when the right fingers performed the flexion-extension exercise. The left primary sensorimotor cortex, left prefrontal area, and both supplementary motor areas were activated with flexion-extension exercise of the left hand. Brain mapping for both APB muscles using TMS showed that no MEP was evoked in the right hemisphere, but a APB total of 5 sites were evoked in the left hemisphere simultaneously. The optimal scalp site for both APB muscles was present at the same site. The MEPs of both muscles which were evoked by stimulation of the optimal scalp site, showed similar latencies, amplitudes, and figures of potential. The similarities in both MEPs and the same optimal scalp site support the assumption that MEPs of both APB muscles are produced by the corticospinal tract originating from the same motor cortex. Our results showed that the ipsilateral motor pathway extended from the unaffected left hemisphere to both hand muscles. This finding may reflect functional reorganization of motor area in a patient with congenital brain disorder. PMID- 11733892 TI - Lower lip-lifting brace for bilateral facial nerve palsy: a case report. AB - A 55-year-old man suffered from pontine hemorrhage 5 years before he visited our outpatient clinic with complaints of gait disturbance and dysphagia. At the first examination, his inability to close his mouth, eyes, and lower lip led to the diagnosis of facial diplegia. He was instructed to wear a gauze surgical mask and to use artificial saliva for his xerostomia. A videofluorogram of his swallowing excluded aspiration but revealed dysphagia attributable to neck hyperextension arising from efforts to prevent food spilling from his mouth. We prescribed a brace to lift his lower lip as a treatment of his dysphagia. This brace covered his chin to support his lower lip. Our brace resulted in improved function; liquids no longer leaked from his mouth and because the lip elevation eliminated his xerostomia, he no longer required artificial saliva or the gauze mask. PMID- 11733893 TI - Isolated subscapularis tear from minimal trauma in a recreational athlete: a case report. AB - A 49-year-old male recreational weight lifter suffered an isolated partial tear of the subscapularis muscle after minimal trauma. His history was atypical for development of a subscapularis tear, which made his case more difficult to evaluate and subsequently treat. His recovery was uncomplicated and was facilitated by a physical therapy program to enhance rotator cuff strength and proprioception. This report includes a brief review of the literature on the etiology and management of these tears. This case emphasizes the importance of an appropriate physical examination and judicious use of imaging techniques so that uncommon disorders, even those with unusual presentations, can be diagnosed and managed to obtain good outcomes. PMID- 11733894 TI - Quantifying fluid intake in dysphagic stroke patients: a preliminary comparison of oral and nonoral strategies. AB - OBJECTIVE: To determine whether dysphagic stroke patients receiving oral (thickened-fluid dysphagia) diets or nonoral (enteral feedings supplemented with intravenous fluids) diets met their estimated fluid requirements. DESIGN: Cohort study. SETTING: University-affiliated hospital. PARTICIPANTS: Thirteen dysphagic patients with new strokes were studied for 21 days postadmission to hospital. INTERVENTIONS: Seven patients (group 1) were started on nonoral feeding and later progressed to oral diets and 6 patients (group 2) received oral dysphagia diets only. MAIN OUTCOME MEASURE: Fluid intake. RESULTS: Fluid intake of patients in group 1 significantly declined over the 21 days (mean +/- standard deviation, 3158 +/- 523mL/d vs 984 +/- 486mL/d; p < .0001), representing 134% +/- 26% and 43% +/- 20% of their fluid requirements, respectively. Mean fluid intake of patients in group 2 was 755 +/- 162mL/d, representing 33% +/- 5% of requirements. This volume was significantly lower than the fluid intake of patients who received nonoral feeding (p < .0001). CONCLUSIONS: Dysphagic stroke patients who received thickened-fluid dysphagia diets failed to meet their fluid requirements whereas patients on enteral feeding and intravenous fluid regimens received ample fluid. PMID- 11733895 TI - The CONSORT statement: Progress in clinical research in pediatric surgery. PMID- 11733896 TI - World Federation of Associations of Pediatric Surgeons. Declaration of pediatric surgery. PMID- 11733898 TI - Intestinal function and metabolism in the early adaptive phase after massive small bowel resection in the rat. AB - PURPOSE: The aim of this study was to investigate the early adaptive responses in metabolism and gut function after massive small bowel resection. METHODS: Male Wistar rats underwent an 80% small bowel resection (Ent group, n = 9) or a transection and reanastomozing (Sham group, n = 7). After 24 hours, substrate fluxes across the gut were determined together with intestinal protein synthesis, polyamine concentrations in gut tissue, and gut function by testing intestinal permeability using the urinary recovery of lactulose and rhamnose. To test for the effect of starvation, healthy starved rats were studied. RESULTS: In the Ent group, intestinal uptake of glucose, lactate, glutamine, phenylalanine, branched chain amino acids, and total amino acids were equal to or higher than that in Sham rats. Intestinal protein synthesis increased, accompanied by an increase in spermidine to spermine polyamine ratios in the ileum and in the jejunal muscular layer. The urinary lactulose to rhamnose ratio also increased, suggesting increased intestinal permeability. CONCLUSIONS: 24 hours after massive small bowel resection, adaptive responses in metabolism and gut function already can be observed, as indicated by increased intestinal uptake of substrates and increased protein synthesis. This, however, is accompanied by an increase in intestinal permeability, which may indicate impaired intestinal barrier function. J Pediatr Surg 36:1746-1751. PMID- 11733899 TI - Results of bowel plication in addition to primary anastomosis in patients with jejunal atresia. AB - BACKGROUND/PURPOSE: Disturbed intestinal transit (DIT) associated with bowel dilation occurs in some postoperative patients with jejunal atresia. Bowel plication (BP) has been introduced to prevent the DIT, but the long-term results of BP are unclear. METHODS: The authors reviewed the preoperative and operative records and postoperative clinical courses (for 2.3 to 7.0 years; mean period, 3.7 years) of 19 jejunal atresia patients, 4 of whom had undergone additional BP with primary anastomosis after dilated bowel resection or tapering jejunoplasty at neonatal surgery. The degree of DIT was evaluated by the clinical symptoms, weight gain, whether reoperation was performed, and duration from the operation to receiving an oral feeding volume of at least 130 mL/kg/d. The patients then were assigned 4 grades (0 to 3). To determine the presence or absence of bowel dilation at the BP site, the plain abdominal x-rays were reviewed. RESULTS: (1) Within 2 months after surgery, 4 patients without BP underwent operation because of severe DIT. The degree of postoperative DIT in patients who had received additional BP at neonatal surgery was less than that in patients without BP (mean grade, 0.50 v. 2.08). (2) The preoperative clinical features, operative method, and postoperative weight gain were almost similar in patients with and without BP. (3) On abdominal x-ray the bowel dilation remained 6 to 12 months after the operation, but was not observed over 1 year after the operation. CONCLUSIONS: In this preliminary study, the addition of BP after tapering jejunoplasty or resection of dilated bowel may be effective in preventing early postoperative DIT. Further study is necessary to evaluate the long-term results of additional BP at neonatal surgery. J Pediatr Surg 36:1752-1756. PMID- 11733901 TI - Total colonic manometry as a guide for surgical management of functional colonic obstruction: Preliminary results. AB - BACKGROUND/PURPOSE: Functional colonic obstruction (pseudo-obstruction) encompasses a broad group of motility disorders. Medical management of colonic pseudo-obstruction is complex and often fails, leading to surgical referral. In most cases (excepting Hirschsprung's disease) the surgeon is unable to precisely localize the area of functional obstruction. Total colonic manometry can directly measure intraluminal pressures and contractile function along the entire length of the colon. The authors propose that total colonic manometry can be used by the pediatric surgeon to guide the timing and extent of surgical therapy in refractory functional colonic obstruction. METHODS: Four patients were evaluated for functional colonic obstruction. All underwent barium enema and rectal biopsy with a diagnosis of Hirschsprung's disease in one patient. All patients underwent colonoscopy and total colonic manometry. Manometric tracings were obtained while fasting, after feeding, and after pharmacologic stimulation both preoperatively (n = 4) and postoperatively (n = 3). RESULTS: Total colonic manometry identified an abrupt end of normal peristalsis in 2 of the non-Hirschsprung's patients (one in the proximal colon and one in the transverse colon). Medical therapy failed in both of these patients, and they underwent diverting ostomy proximal to the loss of normal peristalsis. The third non-Hirschsprung's patient essentially had normal manometry and was able to have her colon decompressed successfully on a laxative regimen. Repeat manometry after colonic decompression showed return of normal peristalsis in 2 of these patients and continued abnormal peristaltic activity in the third. Definitive surgical intervention based on the results of total colonic manometry was performed on the latter. All 3 patients achieved normal continence. A fourth patient had Hirschsprung's disease confirmed by rectal biopsy and underwent a 1-stage neonatal modified Duhamel procedure, which was complicated by postoperative functional obstruction. Manometry showed a lack of peristaltic function beginning in the right colon. An ileostomy was performed, and timing of ileostomy closure was guided by the return of normal colonic peristalsis seen on manometry. CONCLUSIONS: These initial cases show the utility of total colonic manometry in the management of colonic pseudo-obstruction syndromes. In addition to its diagnostic utility, direct measurement of colonic motor activity can be valuable in deciding the need for and timing of diversion, the extent of resection, and the suitability of the patient for restoring bowel continuity. In Hirschsprung's disease, total colonic manometry can potentially be used to determine suitability for primary neonatal pull-through versus a staged approach. J Pediatr Surg 36:1757-1763. PMID- 11733902 TI - Effect of bilirubin in ischemia/reperfusion injury on rat small intestine. AB - PURPOSE: The aim of this study was to determine the effects of bilirubin in experimental small intestinal ischemia/reperfusion (I/R) injury in rats. METHODS: Thirty rats were divided into 5 groups (n = 6). In group S, saline and in group B, bilirubin, 20 mg/kg were infused via the jugular vein without an additional procedure. In groups S-IR, saline, B(1)-IR and B(2)-IR, 10 and 20 mg/kg/h of bilirubin were infused for 2 hours, respectively. In these groups, an I/R procedure was done after infusions by occluding the superior mesenteric artery for 45 minutes followed by 1 hour of reperfusion. After reperfusion, the small intestines were resected for histopathologic and malondialdehyde (MDA) assessments. Mucosal lesions were scored between 0 and 5. Malondialdehyde levels and histopathologic grades were analyzed statistically. RESULTS: Mucosal injury was severe in S-IR (grade 4 to 5), mild in B(1)-IR (grade 0 to 3) and none in B(2)-IR group (grade 0). Grades of group S-IR were higher than those of B(1)-IR and B(2)-IR statistically (P <.05). Tissue MDA levels of the S-IR group were significantly higher than those of B(1)-IR and B(2)-IR groups (U = 36, P <.05). Bilirubin levels correlated inversely with MDA levels (r = -0.94). CONCLUSIONS: Bilirubin effectively prevents intestinal I/R injury in rat. This observation is consistent with the hypotheses regarding bilirubin as an antioxidant, having a role in the body defense. J Pediatr Surg 36:1764-1767. PMID- 11733903 TI - Recurrent congenital diaphragmatic hernia: A novel repair. AB - BACKGROUND/PURPOSE: Repair of recurrent diaphragmatic hernia continues to be a difficult problem. An innovative method using a nonabsorbable polypropylene prosthetic mesh plug placed via the thoracic approach using minimal dissection is presented. METHODS: A retrospective analysis showed 39 children with congenital diaphragmatic hernia (CDH) who underwent repair between January 1997 and March 2000. Five children suffered a recurrence and underwent repair via the thoracic approach using the Bard Marlex Mesh Perfix Plug (C.R. Bard Inc, Billerica, MA). Follow-up was available in all children and ranged from 1 to 33 months (average, 13.8 months). RESULTS: Age at recurrence ranged from 2 to 48 months (average, 14.8 months), and the average time between initial repair and recurrence was 8.2 months (range, 2 to 16 months). There were no recurrences after the transthoracic mesh plug diaphragmatic hernioplasty. One child died of multiple congenital anomalies 6 months after repair. CONCLUSION: The transthoracic repair of recurrent diaphragmatic hernias using a nonabsorbable polypropylene prosthetic mesh plug represents an innovative approach to a difficult problem in which 5 repairs have been accomplished without recurrence in nearly 14 months of follow up. J Pediatr Surg 36:1768-1769. PMID- 11733904 TI - The biomechanical, morphologic, and histochemical properties of the costal cartilages in children with pectus excavatum. AB - BACKGROUND/PURPOSE: The cause of the pectus excavatum (PE) remains unclear, although some results of research have indicated that the disturbance of the sternum or costal cartilage might be responsible for this deformity. But no decisive evidence has been gained. The authors have analyzed the biomechanical, morphologic, and histochemical properties of the cartilage in PE and intend to support the belief that the disturbance of the cartilage might contribute to the development of PE. METHODS: Thirty-eight specimens of the sixth cartilage were obtained at operation for the PE group (aged from 3 to 6 years; mean, 4.2 years). And 28 specimens of the control group (aged from 3 to 6 years; mean, 4.4 years) were gained from routine postmortem examinations in which the cause of death was unlikely to have affected the cartilage. The biomechanical test was carried out in a material testing machine (Shimadzu AG-10TA, Tokyo, Japan). The relation curve of load-deformation in tensile and compressive tests and the curve of load time in the flexuous test were recorded automatically. The values of the ultimate strength and strain were calculated from this relation curve. The specimens also underwent H&E staining. The values of the area, circumference, mean diameter, maximal diameter, and morphologic factor of the cell and the nucleus of the cartilage in superficial and deep area were determined with the help of image analysis software (GT-2 model, China). The superficial zone (SZ) and deep zone (DZ) of the cartilage were examinated with electron microscopy (JEM-100SX, Japan). The distribution and intensity of type II collagen was shown by immunohistochemistry staining and analyzed with the image analysis software (GT-2 model, Huakang Co, Chengdu, China). The extent and distribution of proteoglycan were analyzed after Safranin-O and periodic acid shiff (PAS) staining. RESULTS: The mean strength of the costal cartilage in the experimental group was less than that in the control group in terms of tension, compression, and flexure (P <.05). The shape of the stress-strain curve for tension and compression in the experimental group was different from the control group. The fracture load in the experimental group was less than in the control group in tension (1.5 MPa versus 2.8 MPa) and in compression (.2 MPa versus 8.3 MPa). The time of fracture in experimental group was 30 seconds compared with 38 seconds in control group. No denaturation or necrosis could be found in light microscopical examination. There was no manifestation of hyperplasia or hypoplasia in the costal cartilage of the PE group. In SZ and DZ areas, the pattern and the number of mitochondria, endoplasmic reticulum, and Golgi in the experimental group were the same as the control group in transmission electron microscopy. Furthermore, the distribution and the number of proteoglycan in the 2 groups did not show a significant difference both in SZ and DZ areas. Although the distribution of the collagen in SZ areas was normal, this pattern was disturbed in DZ areas in the experiment group. The results of type II collagen immunohistochemistry examination was concordant with that change. No significant difference between control and experimental group could be seen in Safranin-O and PAS staining for proteoglycan. CONCLUSIONS: The biomechanical stability of the cartilage was decreased in the PE group. This might be caused by the disorderly arrangement and distribution of the collagen in the cartilage of PE patients. J Pediatr Surg 36:1770-1776. PMID- 11733905 TI - Short-term complications of central line placement in children with the human immunodeficiency virus. AB - PURPOSE: The aim of this study was to characterize the perioperative complications of central venous catheter placement in children infected with human immunodeficiency virus (HIV) METHODS: A retrospective chart review was conducted of all central venous catheters placed by the surgical service into HIV infected children from 1988 to 1998 at a large urban children's hospital. Complications occurring within 1 month of catheter placement were analyzed for several host and environmental factors. RESULTS: Forty HIV-positive patients underwent 60 central venous access procedures. Thirty-two of the patients were severely immunosuppressed. Eight catheter placements (13%) resulted in perioperative complications, including hemorrhage (n = 2), site infection (n = 2), catheter sepsis (n = 2), thrombotic occlusion (n = 1), and a pleural effusion secondary to catheter malposition (n = 1). Only 3 patients required catheter removal. There was no significant relationship between either hemophilia or thrombocytopenia and perioperative hemorrhage. No significant relationship was found between infectious complications and preoperative white blood cell count, absolute neutrophil count, CD4% and CD4#, suggesting that a patient's compromised immune status should not be considered a contraindication to central venous catheter placement. CONCLUSION: The complication rate of central venous catheter placement into HIV-infected children is low (<15%), but is still higher than that of the general pediatric population. With careful preoperative preparation this procedure can be performed safely, even in patients with advanced HIV disease. J Pediatr Surg 36:1777-1780. PMID- 11733906 TI - Novel use of an established agent: Topotecan is anti-angiogenic in experimental Wilms tumor. AB - BACKGROUND/PURPOSE: Antiangiogenic agents offer a new approach to the treatment of aggressive neoplasms, yet very few agents are available for current use. The authors have shown previously the efficacy of antiangiogenic therapy in experimental Wilms tumor, using an investigative antibody. They hypothesized that topotecan, administered in a regimen targeting endothelial cells, would suppress tumor growth and angiogenesis in experimental Wilms tumor. METHODS: Experimental tumors were induced in the left kidneys of athymic mice by injection of cultured Wilms tumor cells. Topotecan (0.36, 0.6, 1.0, 2.0, and 3.0 mg/kg) or vehicle was injected intraperitoneally in 2 cycles over a 6-week period. Fluorescein angiograms and platelet endothelial cell adhesion molecule-1 staining of primary tumors were performed to ascertain vascular architecture. Endothelial apoptosis was assessed by TdT-mediated dUTP nick end labeling assay. RESULTS: Tumor weights were reduced significantly in treated versus control animals, even in the lowest dose group. Endothelial cell staining and angiography results showed relatively sparse vascularity in treated xenografts. Endothelial apoptosis was observed in treated but not control tumors. CONCLUSIONS: Topotecan, delivered in an "antiangiogenic" regimen, even at very low doses, significantly inhibited growth of experimental Wilms tumors. No adverse effects were noted at low doses. Thus, the established chemotherapy agent topotecan may be useful in a novel role: as antiangiogenic therapy. J Pediatr Surg 36:1781-1784. PMID- 11733907 TI - The inhibitor of apoptosis protein survivin is associated with high-risk behavior of neuroblastoma. AB - BACKGROUND/PURPOSE: Apoptotic factors inducing or preventing cell death may intrinsically govern the behavior of some tumors. Survivin is a recently described member of the inhibitor of apoptosis protein (IAP) family, that is expressed in a cell cycle-dependent manner and is found in tumors of unfavorable histology. This study examines the presence of several apoptotic factors, including survivin, in neuroblastoma (NB) tumors. Clues to survivin's function in NB are provided by examining its association with behavior and cell dynamics in tumors and cell lines. METHODS: Expression of a panel of apoptosis factors were quantified in 15 NB and related tumors before chemotherapy and in 3 NB cell lines (NB7, NB10, and NB16). Survivin and other apoptotic factors, as well N-myc amplification in primary tumors was correlated with recurrent disease and outcome. Proliferation rate, apoptosis assays, cell cycle analysis, and drug- or immune-mediated cell death were assessed in cell lines and evaluated in the context of differential survivin and apoptosis gene expression. RESULTS: All 7 tumors that went on to recur expressed survivin, whereas expression was absent in all 8 tumors that went into remission. N-myc was amplified in 4 (57.1%) of the 7 recurrent tumors. Of the 8 tumors that were cured, Fas was expressed in 3 (38%), TRAIL-R1 in 6 (75%) and tumor necrosis factor (TNF)-R1 in 8 (100%), whereas these pro-apoptotic receptors were present in only 1 (14%), 1 (14%), and 4 (57%) of the 7 tumors that went on to recur, respectively. Of the 3 cell lines, NB10 expressed the least survivin, displayed the lowest proliferation index, and had the fewest number of cells in the G2/M (mitotic) phase of the cell cycle. Furthermore, NB10 also was most sensitive to TNF-related apoptosis-inducing ligand (TRAIL) or etoposide-induced cell death. CONCLUSIONS: In primary NB tumors, survivin expression was associated with tumors of high risk and unfavorable prognosis, whereas pro-apoptotic receptor expression was more abundant in tumors of favorable prognosis. In this small series, survivin expression appeared to be more predictive of recurrent disease than N-myc amplification. In cell lines, survivin expression was cell cycle dependent, and its expression was associated with greater proliferation rates and greater resistance to drug- or immune mediated cell death. Survivin expression may become a useful prognostic marker in NB and could be a potential target for the treatment of this tumor. J Pediatr Surg 36:1785-1791. PMID- 11733908 TI - Obstructive jaundice secondary to lymphoma in childhood. AB - In children, biliary obstruction secondary to lymphoma is rare. Previous reports in adults and children suggest that these lymphomas are associated with a poor prognosis. The authors reviewed the medical records and imaging studies of 4 children treated for pancreaticobiliary lymphoma at our children's hospital over the past 10 years. All 4 presented with jaundice. Abdominal ultrasound scan and computed tomography (CT) scans were helpful in defining the anatomy of the obstruction. In the cases of involvement of the porta hepatis, the diagnosis was made by biopsy. In one child a mass was in the head of the pancreas, and evaluation of frozen section biopsy results were not diagnostic for lymphoma, and a major resection was performed. All the lymphomas were of nonHodgkin's B-cell type and one was a Burkitt's lymphoma. All responded promptly to chemotherapy. One child had a testicular relapse and currently is receiving additional therapy. Pancreaticobiliary lymphomas are an unusual cause of obstructive jaundice in children. Biopsy alone without resection or biliary drainage is recommended surgical therapy. Long-term survival rate in children with this disorder appears to be more promising than previously reported. J Pediatr Surg 36:1792-1795. PMID- 11733909 TI - Testicular tumors in children. AB - PURPOSE: The aim of this study was to present an updated picture of surgical management of pediatric testicular tumors based on our 30 years' experience, which consisted of one of the largest noncollected series treated in a single medical center. METHODS: Records of children who were treated for testicular tumor in our unit from 1970 to 1999, inclusive, were reviewed retrospectively. Information recorded for each patient included age, sex, past medical history, clinical characteristics, diagnostic procedures, treatment methods, histopathologic findings, and outcome. RESULTS: Fifty-one patients with a mean age of 3.8 +/- 0.5 years were treated for testicular tumors. Of these, 35 (69%) had germ cell testis tumor (GCT) and 16 (31%) had non-germ cell testis tumor (NGCT). Endodermal sinus tumor and paratesticular rhabdomyosarcoma were the dominant histologic subtypes in each group, respectively. The most common mode of presentation was painless scrotal mass. At initial presentation, retroperitoneal (n = 5), both retroperitoneal and lung (n = 2), and retroperitoneal and liver (n = 3) metastases were recorded in 10 (19%) patients. Initial operative procedures were radical inguinal orchiectomy (RIO) (n = 29), scrotal orchiectomy (SO; n = 9), bilateral RIO (n = 2), both RIO and unilateral retroperitoneal lymph node (RPLN) excision (n = 6), testis-sparing enucleation of the tumor (n = 5). SOs were performed elsewhere, and these patients underwent high ligation (n = 4) and both high ligation plus RPLN excision (n = 5) in our unit. Histopathologically, spermatic cord invasion and RPLN involvement were present in 10 patients. Scrotal recurrences were encountered in 2 patients who had scrotal orchiectomy initially. Retroperitoneal recurrences were noted in a patient presenting with stage I embryonal carcinoma and in 2 patients presenting with group IV paratesticular rhabdomyosarcoma. The mean follow-up period was 89 +/- 10 months. Four patients with stage IV embryonal carcinoma (n = 2) and group IV paratesticular rhabdomyosarcoma (n = 2) died of progression of the disease. All remaining patients were alive and disease free at their last outpatient appointment. No significant difference was noted with regard to 5-year survival rates between (1) malignant GCT and paratesticular rhabdomyosarcoma patients (91% v 80%) and (2) patients treated by RIO (88%), SO plus high ligation (87%), and RIO plus RPLN excision (80%). Five-year survival rates were 100% for stage I, II, III patients and 33.3% for stage IV and group IV patients presenting with malignant testicular tumors (P <.05). CONCLUSIONS: Childhood testicular tumors deserve special attention from the therapeutic point of the view. A solid scrotal mass should be considered malignant until proved otherwise. Any suspicion of the testicular tumor warrants an inguinal approach to prevent scrotal violation by the tumor. Current trends emphasize that testis-sparing surgery should be performed for benign lesions such as teratoma, leydig cell tumor, and epidermoid cyst based on frozen biopsy findings. Literature findings and our experience suggest that RIO is the accurate treatment for stage I malignant GCT and group I and IIa paratesticular rhabdomyosarcoma. RPLN excision is not of benefit either as a staging or therapeutic procedure in stage I and group I and IIa diseases of these tumors. RPLN excision should be reserved for (1) malignant GCT patients who have persistent elevation of alpha-fetoprotein after orchiectomy in the presence of normal total body CT scan, and for patients presenting with stage II and III disease with definitive abnormality on CT scans, and (2) group IIb, IIc, and III paratesticular rhabdomyosarcoma patients with radiologic evidence of retroperitoneal involvement on CT scans. High ligation should be done as a complementary procedure after SO to increase the survival rates. J Pediatr Surg 36:1796-1801. PMID- 11733910 TI - Impact of mentor guidance in surgical career selection. AB - PURPOSE: The aim of this study was to survey graduates of a university general surgical residency training program to determine factors that influenced their selection of a specialty field. METHODS: A 39-item questionnaire was mailed to 86 graduates of a university general surgery program who matriculated from 1975 to 1989. The impact of lifestyle, technology, clinical opportunity, and mentor guidance in the residents' selection of a specialty field and eventual clinical practice was assessed. Results were analyzed using Fisher's Exact test with significance determined at P less than.05. RESULTS: The response rate was 65% (56 of 86). Eighty percent of respondents identified the most important aspect influencing their choice of specialty was interest in that field; additional factors included perception of prestige, presence of clinical opportunity, mentor influence, and family priorities. Sixty-six percent of respondents chose the same career as their mentor, attributing this to the mentor's skill (n = 36, 68%), achievements (n = 35, 66%), and verbal recommendations about their specialty field (n = 24, 45%), p < 0.05. CONCLUSIONS: Surgical residents use many criteria in selecting a field of specialty with mentor guidance an important component. Knowledge of these influential areas should help training programs offer appropriate career guidance. J Pediatr Surg 36:1802-1804. PMID- 11733911 TI - Experimental small bowel transplantation using a newborn intestine in rats: IV. Effect of cold preservation on graft neovascularization. AB - BACKGROUND/PURPOSE: University of Wisconsin (UW) solution is one of the most superior organ preservation solutions for liver, kidney, and pancreas; however, it still is controversial for intestinal preservation. Here, the authors studied the efficacy of preservation with 2 kinds of solutions, UW and modified TOM (m TOM) solutions in our experimental newborn intestinal transplantation model. UW solution was used as a standard intracellular and m-TOM solution as an extracellular preservation solution. Lactated ringer (LR) solution was used as a control. METHODS: Newborn intestine, which were preserved in these solutions for 24 or 48 hours, were transplanted in the subcutaneous spaces of the syngeneic recipients without surgical vascular anastomosis and histologically examined 14 days after grafting. The preserved grafts were evaluated histologically by use of light and electron microscopy just after preservation. The biochemical parameters such as LDH and serotonin also were measured in the supernatants of preservation solutions. RESULTS: Fresh newborn grafts were revascularized successfully at a rate of 80% (16 of 20). After 24 hours of preservation, 65% (13 of 20), 75% (15 of 20), and 85% (17 of 20) of the grafts were observed to be revascularized in LR, m-TOM, and UW solutions, respectively. After 48 hours of preservation, 60% (12 of 20), 80% (16 of 20), and 80% (16 of 20) of the grafts also were revascularized in the respective solutions (no statistic difference among the groups). The cold-preservation did not affect the neovascularization of newborn intestine until 48 hours. Histologic findings of the preserved intestine and biochemical analyses showed that UW and m-TOM solutions kept villous architectures of the preserved grafts, however, might be harmful to enterochromaffin cells. CONCLUSION: Long-time preservation of newborn intestine did not interfere with neovascularization and maturation. J Pediatr Surg 36:1805 1810. PMID- 11733912 TI - Intestinal damage in gastroschisis correlates with the concentration of intraamniotic meconium. AB - BACKGROUND/PURPOSE: Contact with amniotic fluid (AF) causes intestinal damage in gastroschisis, which has been shown to be caused by intraamniotic meconium. However, whether this intraamniotic meconium-induced intestinal damage is concentration dependent has not been investigated previously. The purpose of this study is to investigate the effects of intraamniotic human meconium at various concentrations on the intestines of chick embryo with gastroschisis. METHODS: Five-day-old fertilized chick eggs were used. Gastroschisis was created through the amniotic cavity without opening the allantoic cavity. Sterile meconium was obtained from newborn humans. Meconium suspensions at various concentrations were prepared using saline and instilled into the amniotic cavity. RESULTS: Intraamniotic 1:200 and 1:400 meconium was found to cause intestinal damage. Meconium concentrations lower than 1:400 did not cause intestinal damage. Histopathologic examination of the intestines of the 1:200 and 1:400 meconium groups showed serosal thickening, inflammation, focal fibrin, and collagen deposits. Histopathologic features of the intestines of the 1:600 and 1:800 meconium groups did not differ from the intestines of the control group. CONCLUSION: Intraamniotic meconium, which is responsible for intestinal damage in gastroschisis, must reach a threshold level to induce intestinal damage. J Pediatr Surg 36:1811-1815. PMID- 11733913 TI - Primary transanal rectosigmoidectomy for Hirschsprung's disease: Preliminary results in the initial 33 cases. AB - PURPOSE: The authors describe their newly developed technique-primary transanal rectosigmoidectomy for Hirschsprung's disease (HD) and its preliminary results in neonates and infants. METHODS: Thirty-four consecutive patients (26 boys) with biopsy-proven rectosigmoid HD, aged 18 days to 4 years, underwent this new procedure. Rectal mucosectomy started 1 to 1.5 cm posteriorly and 2 to 3 cm anteriorly proximal to the dentate line. The rectal muscular sleeve below the peritoneal reflection was resected to the level of the striated muscle complex, leaving a shorter muscular cuff, into which a partial internal sphincterotomy was made posteriorly. An oblique anastomosis was constructed between the pull-through ganglionic colon and the anus canal. RESULTS: The mean time for the operation was 160 minutes, and the average length of bowel resected was 29.5 cm (range, 12.5 to 41 cm). Two children (6.06%, 2 of 33) had 2 to 5 episodes of postoperative enterocolitis (EC). One was cured by rectal irrigation and dilation, and the another by Lynn's myectomy. Eighty-four percent of patients had 1 to 6 bowel movements per day during a 6- to 18-month follow-up period. CONCLUSIONS: Primary transanal rectosigmoidectomy for HD is logical and associated with excellent early results. A long-term follow-up is required to determine bowel functions. J Pediatr Surg 36:1816-1819. PMID- 11733914 TI - Postoperative pneumatosis intestinalis in infants does not automatically preclude enteral feeding. AB - BACKGROUND: A degree of feed intolerance after neonatal abdominal surgery is common but in an otherwise well baby enteral feeding usually is continued at the highest tolerated level. However, the presence of rectal bleeding, pneumatosis intestinalis, or portal vein gas seen on plain abdominal x-rays suggest the possibility of postoperative necrotising enterocolitis. When this happens feedings usually are stopped for 7 to 10 days, and intravenous antibiotics and total parental nutrition are commenced. METHODS: The authors report 12 episodes of rectal bleeding and 11 episodes of pneumatosis intestinalis in 3 infants who previously had undergone neonatal abdominal surgery for intestinal malformations. In 7 of these episodes, feedings were neither stopped nor were antibiotics given. At the time of these 7 episodes, the infants were more than 3 kg in weight, had no significant cardiac or respiratory pathology, were all clinically stable, had no evidence of peritonitis, had no thrombocytopenia, and were greater than 37 weeks postconception. RESULTS: The 3 infants were monitored closely. There were no early or late problems observed attributable to this management. CONCLUSION: Carefully selected clinically stable patients that have postoperative pneumatosis intestinalis or exhibit rectal bleeding may be successfully managed by reduced enteral feedings with no antibiotics. J Pediatr Surg 36:1820-1823. PMID- 11733915 TI - Effects of tezosentan, a dual endothelin receptor antagonist, on the cardiovascular and renal systems of neonatal piglets. AB - BACKGROUND/PURPOSE: Endothelin is a potent biological vasoactive mediator in the cardiovascular and renal systems. Little is known of the effects of endothelin antagonism on the developing heart and kidney, and we hope to show that endothelin does have an important role in the cardiovascular and renal systems of the developing neonate. In this study the authors have examined the effects of tezosentan, a nonselective endothelin-A and endothelin-B receptor antagonist designed for parenteral use, on the cardiovascular and renal systems of healthy neonatal piglets. METHODS: Eight, 7- to 10-day old domestic piglets weighing 2.5 to 3.0 kg were anesthetized, intubated, and ventilated with catheters placed into the jugular vein, left ventricle, and femoral artery. Urine output was monitored via a suprapubic cystostomy. After baseline data were obtained the piglets received tezosentan (1 mg/kg/h) for 1 hour. A set of data was collected just before discontinuation of the 1-hour infusion of tezosentan and another set was collected 1 hour after the discontinuation of the drug. Mean arterial pressure (MAP), heart rate (HR), and urine output (UV) were monitored continuously and cardiac index (CI), systemic vascular resistance (SVR), renal blood flow (RBF), and renal vascular resistance (RVR) were calculated from gamma counts obtained from injections of radio-labeled microspheres at end of the different time periods. Glomerular filtration rate (GFR) was obtained by the sodium iothalamate method. Data were averaged and plotted versus time and analyzed statistically by a Student's t test. RESULTS: (P <.05 versus baseline*). In our experimental animals the infusion of tezosentan diminished MAP and SVR from baseline values of 94 +/- 7 mm Hg and 0.14 +/- 0.03 mm Hg/mL/min, respectively to TEZO values of 62 +/- 4* mm Hg and 0.07 +/- 0.02* mm Hg/mL/min. CI increased from 278 +/- 58 to 367 +/- 75* mL/min/kg with tezosentan. There also was a statistically significant increase in RBF from 1.16 +/- 0.38 to 1.86 +/- 0.37* mL/min/kg, an increase in UV from 0.57 +/- 0.24 to 0.64 +/- 0.12* mL/min, a decrease in RVR from 4.60 +/- 1.47 to 2.03 +/- 0.36* mm Hg/mL/min, and no change in the GFR. CONCLUSIONS: The inhibition of endothelin receptors with tezosentan produced a statistically significant effect on the piglet cardiovascular system with a drop in MAP and SVR and an increase in CI and HR. It also produced a statistically significant increase in RBF and UV and a decrease in RVR without affecting GFR. J Pediatr Surg 36:1824-1828. PMID- 11733916 TI - Urethral catheterization in hypospadias surgery: Should the device enter the bladder or be made a urethral stent? AB - BACKGROUND: Although bladder catheterization causes contractions, accidental removal and urinary retention are risks of stents in hypospadias repair. METHODS: An 8F feeding tube was used as a stent in 22 patients (group I). In the other 22 patients (group II), the same size catheter was passed into the bladder, and oxybutynin chloride was administered. The elapsed time to first voiding, pain, and straining was observed in group I and the groups were compared regarding time of catheter removal, accidental removal of catheter/stent, hospital stay, and complications. RESULTS: All stented patients strained at first voiding. Nineteen showed pain, and only 5 voided in the first 8 hours. The catheterized group had no such problems. Time of catheter/stent removal and hospital stay were similar. Three stents were dislodged in patients with significant straining. Meatal stricture was noticed in 5 and 3 patients and fistula in 5 and 2 patients of group I and II, respectively. Complications were significant in patients whose stents were removed accidentally. CONCLUSIONS: Stenting led to significant patient irritability with voiding problems. The authors believe that this caused displacement and accidental removal of the stent and eventually resulted in meatal stenosis and fistula. The findings show that bladder catheterization prevents these complications. J Pediatr Surg 36:1829-1831. PMID- 11733917 TI - Leiomyoma at the site of esophageal atresia repair. AB - Esophageal leiomyomas are rare in the pediatric population. They frequently occur in association with other anomalies, such as Alport's syndrome, osteoarthropathy, and leiomyomas elsewhere in the body. The authors describe the case of a focal esophageal leiomyoma in a 12-month-old girl with a history of long-gap esophageal atresia. The patient initially underwent bouginage of the proximal pouch while awaiting definitive repair. After esophageal repair, the patient required multiple dilatations for anastomotic strictures. A segmental resection of the esophagus was performed because of recurrent strictures. A leiomyoma, arising from the site of the previous esophageal atresia repair, was noted on histologic evaluation. Esophageal leiomyomas have not been reported previously in a child with esophageal atresia. J Pediatr Surg 36:1832-1833. PMID- 11733918 TI - Closed gastroschisis: Antenatal and postnatal features. AB - Four cases of antenatally diagnosed gastroschisis are described in whom there was significant closure of the abdominal wall defect around the prolapsed midgut. In 2 cases this resulted in near-total midgut infarction and short bowel syndrome. In a further case there was entry and exit jejunal and colonic atresia and significant midgut damage but some viability of extrabdominal bowel. In the final case, although there was complete closure of the ring around the base of the midgut, no intestinal loss had occurred. In the latter 2 cases, serial antenatal ultrasound imaging had shown development of intrabdominal bowel dilatation prompting early preterm delivery. Three of these infants currently are alive and well. The authors suggest the use of the term closed (or closing) gastroschisis to describe this phenomenon. J Pediatr Surg 36:1834-1837. PMID- 11733919 TI - Secondary sclerosing cholangitis and portal hypertension after O157 enterocolitis: Extremely rare complications of hemolytic uremic syndrome. AB - The authors experienced an extremely rare case of secondary sclerosing cholangitis and portal hypertension developed as late complications of hemolytic uremic syndrome (HUS) owing to Escherichia coli O157:H7 in a 2-year-old boy. HUS after E coli O157 infection is the most frequent cause of acute renal failure in childhood and occasionally is accompanied by extrarenal complications such as encephalopathy, cardiomyopathy, ischemic colitis, and pancreatitis. Rarely, late colonic stenosis may develop secondary to the ischemic damage. Sclerosing cholangitis and subsequent cirrhosis with portal hypertension are very uncommon as late complications of HUS. To our knowledge, such a case has not been previously reported in the literature. J Pediatr Surg 36:1838-1840. PMID- 11733920 TI - Familial presacral masses: Screening pitfalls. AB - BACKGROUND/PURPOSE: Anterior sacral meningocele (ASM) and presacral teratoma (PT) are related malformations. Both can have familial occurrence. Surgical treatment of occult cases is essential. Hence, it is important to screen the asymptomatic relatives. Plain radiograph of the sacrum to look for any bony abnormalities is the usual accepted screening modality. The authors report ASM and PT in siblings (with positive family history) with no sacrococcygeal bony abnormality on imaging. Both the siblings had presacral mass on imaging. METHODS: The medical records of 2 siblings with ASM and PT were reviewed for the clinical presentation, diagnostic workup, and family history. RESULTS: One sibling was symptomatic, whereas the other was completely asymptomatic. A family history of sacral bony abnormalities and anterior meningoceles was present in father, 2 paternal cousins, paternal uncle, paternal aunt, paternal grandmother, and paternal grand uncle. CONCLUSIONS: These case reports suggest that imaging for screening for ASM and PT should be directed at identifying the presacral mass rather than sacral bony defect. Hence, computed tomography or magnetic resonance imaging looking for presacral mass rather than plain radiograph should be the screening modality. J Pediatr Surg 36:1841-1844. PMID- 11733921 TI - Pulmonary epithelial cell maturation in hyperplastic lungs associated with fetal tracheal agenesis. AB - BACKGROUND/PURPOSE: Cellular differentiation of pulmonary hyperplasia has not been reported in human cases. The authors studied surfactant protein expression and ultrastructure of pulmonary epithelial cells in fetal hyperplastic lungs associated with congenital tracheal agenesis. METHODS: The maturation of pulmonary epithelial cells was assessed by immunohistochemical examination for surfactant proteins (SP-A, mature SP-B, proSP-B, proSP-C, and SP-D) and transmission electron microscopy. As controls normal lung portions of 8 fetuses born at 21 weeks gestation were used. RESULTS: Mature SP-B and SP-D was detected in terminal airways in this case, but not in controls. In electron microscopy, lamellar bodies were recognized, and glycogen granules were less abundant in terminal airway cells. CONCLUSION: The differentiation of pulmonary epithelial cells appeared to be more advanced for the gestational age in pulmonary hyperplasia with congenital tracheal agenesis. J Pediatr Surg 36:1845-1848. PMID- 11733922 TI - Acute abdomen caused by Salmonella typhimurium infection in children. AB - Salmonella spp. infections can be particularly challenging when they manifest as acute abdominal problems and lead to emergency surgery. Examples of such serious conditions are Salmonella-related intestinal perforation, gallbladder involvement, salpingitis, and peritonitis. Mesenteric lymphadenitis associated with Salmonella typhimurium mimics acute appendicitis and can make it difficult to establish a timely and definitive diagnosis in young patients who present with right lower abdominal pain. Paralytic ileus is a fairly common manifestation of Salmonella infection at all ages, but complete intestinal obstruction requiring surgical intervention is very rare. Because of the nature of the diagnostic process, a significant number of patients with Salmonella infection present with acute abdomen and undergo needless operations. This report describes the cases of 2 pediatric patients who underwent surgery to address persistent pain in the right lower abdominal quadrant and complete intestinal obstruction, respectively. The first patient had inflamed mesenteric lymph nodes that caused appendicitislike symptoms, and the second had dense adhesions between the mesentery and the terminal segments of the ileum that led to intestinal blockage. Serology results showed that both patients' titers for BO ("B and O agglutinating [BO]") antibodies rose to 1:640 in the week after their admission to hospital, a pattern and level that is indicative of S typhimurium infection. J Pediatr Surg 36:1849-1852. PMID- 11733923 TI - An unusual late complication of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) now is managed frequently successfully without surgical intervention. NEC may result in strictures, which present after the acute inflammatory process has resolved. Strictures usually present as obstruction in the first year or two of life. A case report is presented of an 11 year-old child who had symptoms from a previously undiagnosed NEC stricture as a result of pica when coins obstructed the stricture. As treatment of NEC continues to improve, more and later complications of this disease can be expected. J Pediatr Surg 36:1853-1854. PMID- 11733924 TI - Cystic Meckel's diverticulum: A rare cause of cystic pelvic mass presenting with urinary symptoms. AB - An atypical form of Meckel's diverticulum is presented. A 5-year-old girl was admitted with recurrent urinary tract infections. Ultrasonography showed a cystic mass, which pressed and displaced the urinary bladder on its posterosuperior aspect. Laparotomy results showed a cystic mass originating from the antimesenteric border of the ileum, which was attached tightly to the urinary bladder. It was diagnosed as a cystic Meckel's diverticulum, which had undergone abscess formation with preservation of the ileal mucosa in some areas and had severe inflammatory changes on its wall. The case is unique both in the mode of presentation and type of complication. J Pediatr Surg 36:1855-1858. PMID- 11733925 TI - Unusual presentation of psoas abscess in a child. AB - Primary abscess of the psoas muscle is relatively rare in infants and young children. The clinical presentation of the disease, with limping, fever, and abdominal pain, may be confused with conditions such as septic arthritis of the hip, osteomyelitis, or appendicitis. The authors present an unusual case of a ruptured left psoas abscess presenting as generalized peritonitis in a child. J Pediatr Surg 36:1859-1860. PMID- 11733926 TI - Colon interposition for esophageal stenosis in a patient with epidermolysis bullosa. AB - Epidermolysis bullosa (EB) is a disease with 3 forms, most hereditary, characterized by spontaneous blistering lesions. The autosomally inherited form, epidermolysis bullosa dystrophica recessive (EBDR), is responsible for esophageal lesions consisting of web or stenosis. The authors could find only 9 cases treated by various esophageal replacement procedures in the literature, and the experience with 1 case treated by colon interposition is presented. J Pediatr Surg 36:1861-1863. PMID- 11733927 TI - The use of dipyridamole to wean from inhaled nitric oxide in congenital diaphragmatic hernia. AB - A full-term neonate with a left-sided congenital diaphragmatic hernia (CDH) was ventilated mechanically by high-frequency oscillatory ventilation (HFOV). Despite inhaled nitric oxide (iNO) at a dose of 15 ppm, the neonate had severe respiratory acidosis and was placed on extracorporeal membrane oxygenation (ECMO) for 2 days. On day 7 of life, surgical repair of the CDH was performed. After the intervention, iNO (20 ppm) had to be restarted because of severe pulmonary hypertension (PHT). Ventilatory support and iNO then were weaned progressively. However, each daily attempt to discontinue iNO (from 2 ppm to 0 ppm), led to severe desaturation with significant right-to-left shunting. At the age of 33 days, dipyridamole (persantin) was administered intravenously at a dose of 0,4 mg/kg/min over 10 minutes and repeated every 12 hours for a total of 3 doses. After the second administration of dipyridamole, iNO could be stopped without rebound of PHT, and the neonate was extubated 1 week later. The authors report the use of dipyridamole for successful withdrawal of iNO. By inhibition of phosphodiesterase type 5, dipyridamole has the potential to increase the level of cyclic guanosine monophosphate in vascular smooth muscle cells, permitting vasodilation and restoration of endogenous NO. J Pediatr Surg 36:1864-1865. PMID- 11733928 TI - Anterior abdominal wall malignant peripheral nerve sheath tumor in an infant. AB - Malignant peripheral nerve sheath tumors (MPNST), also known as malignant schwannomas, are highly malignant lesions that tend to arise in the head and neck region or on the extremities, and only rarely in the retroperitoneum. These tumors are not common in children. Almost 50% of cases are associated with neurofibromatosis. When these neoplasms arise in the retroperitoneum, they usually are difficult to diagnose and to treat. Although various radiologic imaging methods are helpful for identifying some features of the mass, definitive diagnosis requires histologic examination and immunohistochemical staining. After treatment, the tumor recurs in 25% of patients. Five-year survival rates as high as 80% have been reported. Total excision, lack of invasion of surrounding tissues and vessels, and absence of neurofibromatosis, are features associated with better outcome. The authors present the case of a 1-month-old boy who had malignant schwannoma diagnosed in a unique location over the peritoneum of the anterior abdomen. The location and rapid growth of the tumor permitted early identification and total excision. J Pediatr Surg 36:1866-1868. PMID- 11733929 TI - Intraatrial and intracaval Wilms' tumor. PMID- 11733930 TI - Re: Ciftci et al "Pheochromocytoma in Children" (J Pediatr Surg 36:447-452, 2001). PMID- 11733932 TI - Re: Krstic et al "Surgical treatment of the Mullerian duct remnants". PMID- 11733933 TI - Video-assisted thoracoscopic surgery (VATS) in the management of complicated pneumonia in children. PMID- 11733934 TI - Paraneoplastic pemphigus and bronchiolitis obliterans associated with a mediastinal mass: A rare case of Castleman's disease with respiratory failure requiring lung transplantation. AB - Castleman's disease is an infrequent and usually benign lymphoproliferative disorder. Resection of the tumor usually is curative. The immunostimulatory nature of the tumor can, in rare instances, result in paraneoplastic manifestations. The authors present a case of a 14 year old with mucocutaneous ulcerations and progressive dyspnea that was found to have a large mediastinal mass and circulating autoantibodies that were responsible for his paraneoplastic pemphigus and bronchiolitis obliterans. In spite of aggressive immunotherapy to control the autoimmune mucocutaneous lesions, the pulmonary fibrosis was irreversible and progressed to pulmonary failure necessitating lung transplantation. J Pediatr Surg 36:E22. PMID- 11733935 TI - Outcome of paratesticular involvement in infants with neuroblastoma. AB - The authors report on 3 infants suffering from disseminated neuroblastoma (NB) involving the testes or paratesticular structures. INSS stage 4 in 2 cases, and "biological" INSS stage 4S were considered, respectively. One patient with a stage 4 NB died of tumor progression; one patient is under therapy. The patient with NB 4S was cured with preservation of both testes after antineoplastic chemotherapy and reduction of the retroperitoneal primary. J Pediatr Surg 36:E23. PMID- 11733936 TI - Ion-dependence of Z-line and M-line response to calcium in striated muscle fibres in rigor. AB - The calcium-dependent contraction of vertebrate skeletal muscle is thought to be primarily controlled through the interaction of the thick and thin filaments. Through measurement of the Donnan potential, we have shown that an electrical switching mechanism (sensitive to both anions and cations) is present in both A- and I-bands [1]. Here we show that this mechanism is not confined to the contractile apparatus and report for the first time the presence of M-line potentials. The Z-line responds to Ca2+ ions in a similar manner to the A-band under the same solution conditions (phosphate-chloride and imidazole buffers), even though it has no reported Ca2+ binding sites. Z-line potentials were not observed in tris-acetate buffer. The M-line has a markedly different response to any of the other subsarcomeric regions, however, and can only be detected in the phosphate-chloride buffer. Preliminary observations of the M-line potential in creatine kinase-deficient mouse muscle (phosphate-chloride buffer) reveal significant differences in the calcium-induced transitions between two of the genotypes and demonstrate definitively that it is the M-line potential that is being recorded. From these results, it seems likely that the charge response of the Z-line and M-line is being mediated by titin in an anion-dependent manner. Our evidence comes from several observations. First, the similarity between the response of the Z-line potentials to the A-band potentials, where titin is the only link between these structures and second, the differential observation of M line and Z-line potentials in a range of buffers containing different anion(s). Both Z-line and M-line potentials were seen in phosphate-chloride buffer, but only the Z-line potentials could be detected in chloride-only (imidazole) buffer and neither was observed in the acetate buffer. The latter observations can be attributed to two sources. The first is the effect of acetate buffer on the conformation of myosin [2]; the second is the absence of binding of the M-line protein, myomesin, to titin in the absence of phosphate ions [3]. PMID- 11733937 TI - Selective, high-resolution fluorescence imaging of mitochondrial Ca2+ concentration. AB - We have developed a digital image processing technique based on highpass filtering of microfluorimetric images for selective transmission of fine image details corresponding to mitochondria. This technique enabled the detection of the mitochondrial calcium signals with high selectivity, simultaneously with the cytosolic calcium signal. The validity of this technique was supported in primary cultures of rat brain capillary endothelial cells loaded with X-rhod-1 by the results that (i) inhibition of the mitochondrial Ca2+ uptake by discharging the mitochondrial membrane potential selectively abolished the transient of the highpass filtered signal evoked by ATP, and (ii) CGP-37157, a selective blocker of the mitochondrial Na+/Ca2+ exchanger, increased the peak amplitude of highpass filtered (mitochondrial) Ca2+ transients and caused a sustained plateau. The highpass filtering technique enabled the analysis of the mitochondrial Ca2+ transients in high temporal resolution. We found a uniform and monophasic rise of [Ca2+] in the mitochondrial population of the cell, following the cytosolic [Ca2+] with a delay at onset and peak. The introduced highpass filtering technique is a powerful tool in the high spatial and temporal resolution analysis of mitochondrial calcium transients, and it could be especially important in specimens where genetically targeted probes fail. PMID- 11733938 TI - The inositol trisphosphate receptor antagonist 2-aminoethoxydiphenylborate (2 APB) blocks Ca2+ entry channels in human platelets: cautions for its use in studying Ca2+ influx. AB - It has been reported that store-mediated Ca2+ entry (SMCE) in human platelets is likely to be mediated by a secretion-like coupling mechanism. Recently, 2 aminoethoxydiphenylborate (2-APB) has been used in the investigation of SMCE. Here, the mechanism of action of 2-APB is investigated in human platelets. In a Ca2+-free medium (EGTA added), addition of 0.1 U/ml thrombin caused an elevation in [Ca2+]i. Preincubation with 100 microM 2-APB for 170s abolished the release of internal Ca2+. In platelets whose internal Ca2+ stores had been depleted by treatment with 200 nM thapsigargin, addition of extracellular Ca2+ caused an elevation in [Ca2+]i indicative of SMCE. Preincubation with 2-APB decreased SMCE by 95.5+/-1.1%. After activation of SMCE, addition of 2-APB rapidly decreased [Ca2+]i to basal levels; in contrast, the coupling between Trp1 and IP3RII, which has been shown to play an important role in SMCE in platelets, remained intact at the same time points. The rate of decrease of [Ca2+]i and the absence of measurable latency in the effect of 2-APB were comparable to the effects of La3+ (a cation channel blocker). These data suggest that 2-APB may act as a blocker of Ca2+ permeable plasma membrane channels. These data provide further information regarding the mechanism and site of action of 2-APB and highlight the necessity of careful interpretation of work performed using this molecule. PMID- 11733939 TI - Iminocoumarin-based low affinity fluorescent Ca2+ indicators excited with visible light. AB - A series of iminocoumarin-based fluorescent Ca2+ indicators were synthesized and the spectral profiles of their free and Ca2+ bound forms were studied. The newly synthesized compounds incorporate the Ca2+ chelating structure of BAPTA. The chromophore moieties are iminocoumarins substituted at the 3-position with benzothiazolyl, benzoxazolyl and benzimidazolyl groups. These compounds are excited with visible light and their Ca2+ dissociation constants range from 5.4 to 27.5 microM. Fluorescence spectra studies of these probes indicated a clear shift in their excitation wavelength maxima upon Ca2+ binding along with changes in fluorescence intensity that enable the compounds to be used as low Ca2+ affinity, visible excitable probes. PMID- 11733940 TI - Kinetics of inhibition of the plasma membrane calcium pump by vanadate in intact human red cells. AB - The lack of specific inhibitors of the plasma membrane Ca2+ pump (PMCA) has made vanadate (VO3-), a non-specific inhibitor, an invaluable tool in the study of PMCA function. However, three important properties of vanadate as an inhibitor of the PMCA in intact cells, namely its speed of action in different experimental conditions, the reversibility of its inhibitory effects at different doses, and its dose-response, had never been characterized, despite extensive use. We report here the speed, reversibility and dose-response of PMCA inhibition by vanadate in intact human red cells. Near maximal inhibitory concentrations (1mM) in the red cell suspension blocked almost instantly the uphill Ca2+ extrusion by the PMCA, regardless of the intracellular Ca2+ concentration, cation composition of the external media, membrane potential or volume-stability of the cell. PMCA inhibition by vanadate, at concentrations of 10mM and 1mM, was not reversed by washing, resuspending, and incubating the cells for up to 2h in vanadate-free media. Vanadate inhibited PMCA-mediated Ca2+ efflux in intact red cells with a K1/2 of approximately 3 microM, a value similar to that described for the Ca2+ ATPase in isolated red cell membranes. PMID- 11733941 TI - Bcl-2 increases emptying of endoplasmic reticulum Ca2+ stores during photodynamic therapy-induced apoptosis. AB - Photodynamic therapy (PDT) is clinically approved for the treatment of several types of cancer as well as age-related macular degeneration, the leading cause of blindness in the elderly. PDT using the photosensitizer verteporfin has been previously shown to induce rapid apoptosis via a mitochondrial-caspase activation pathway. The impact of PDT on other cellular organelles such as the endoplasmic reticulum (ER) is undefined. The effect of PDT on intracellular Ca2+ ([Ca2+]i) in control and Bcl-2-overexpressing HeLa cells was assessed. A greater [Ca2+]i transient was observed for Bcl-2 overexpressing cells in response to PDT. The PDT induced Ca2+ release was due to the emptying of Ca2+ from ER and possibly mitochondrial stores and was not due to an influx of Ca2+ from the medium. For Bcl-2-transfected cells, the release of Ca2+ was incomplete as determined by a further [Ca2+]i transient produced by the addition of the Ca2+ ionophore ionomycin after PDT. Furthermore, extrusion of Ca2+ was not hindered while ER mediated sequestration of Ca2+ was impaired after PDT. Impairment of ER-mediated sequestration of Ca2+ may be due to the immediate caspase-independent depletion of sarco/endoplasmic reticulum Ca2+ ATPase-2 (SERCA2) that occurred in response to PDT in birth HeLa/Neo and Bcl-2 overexpressed HeLa cells. In summary, PDT induced the rapid degradation of SERCA2 and release of ER and mitochondrial Ca2+ stores. Although overexpression of Bcl-2 did not protect against SERCA2 degradation, it may influence the release of Ca2+ from ER and mitochondrial stores in PDT-treated cells. PMID- 11733942 TI - Regulation by endothelin-1 of Na+-Ca2+ exchange current (I(NaCa)) from guinea-pig isolated ventricular myocytes. AB - The cardiac Na+-Ca2+ exchanger participates in Ca homeostasis, and Na+-Ca2+ exchanger-mediated ionic current (I(NaCa)) also contributes to the regulation of cardiac action potential duration. Moreover, I(NaCa) can contribute to arrhythmogenesis under conditions of cellular Ca overload. Although it has been shown that the peptide hormone endothelin-1 (ET-1) can phosphorylate the cardiac Na+-Ca2+ exchanger via protein kinase C (PKC), little is known about the effect of ET-1 on I(NaCa). In order to examine the effects of ET-1 on I(NaCa), whole cell patch clamp measurements were made at 378C from guinea-pig isolated ventricular myocytes. With major interfering currents inhibited, I(NaCa) was measured as the current sensitive to nickel (Ni; 10mM) during a descending voltage ramp. ET-1 (10 nM) significantly increased I(NaCa) ( approximately 2-fold at -100 mV). Application of a PKC activator (PMA; 1mM: phorbol 12-myristate 13 acetate), mimicked the effect of ET-1. In contrast, the PKC inhibitor chelerythrine (CLT, 1mM) abolished the stimulatory effect of ET-1. An inactive phorbol ester, 4-alpha-phorbol-12,13-didecanoate (4a-PDD, 1mM) had no effect on I(NaCa). Collectively, these data indicate that ET-1 activated I(NaCa) through a PKC-dependent pathway. In additional experiments, isoprenaline (ISO; which has also been reported to activate I(NaCa) ) was applied. The increase in I(NaCa) density with ISO (1mM) was similar to that induced by ET-1 (10nM). When I(NaCa) was pre-stimulated by ET-1, application of ISO elicited no further increase in current and vice versa. ISO also had no additional effect on I(NaCa) when the cells were pretreated with PMA. Application of CLT did not alter the response of I(NaCa) to ISO. We conclude that ET-1 stimulated ventricular I(NaCa) via a PKC dependent mechanism under our recording conditions. Concentrations of ET-1 and ISO that stimulated I(NaCa) to similar extents when applied separately were not additive when co-applied. The lack of synergy between the stimulatory effects of ET-1 and ISO may be important in protecting the heart from the potentially deleterious consequences of excessive stimulation of I(NaCa). PMID- 11733943 TI - The pathogenesis of advanced cervical cancer provides the basis for an empirical therapeutic vaccine. AB - The pathogenesis of carcinogenic human papillomavirus (HPV) infections of the cervix includes early induction of peripheral tolerance of tissue-infiltrating lymphocytes and an imbalanced Th2 response to HPV early virus proteins. As lesions become progressively dysplastic, major histocompatibility complex (MHC)-1 molecules are down-regulated on the surface of abnormal keratinocytes. When the target of MHC-1 class-restricted cytotoxic lymphocytes disappears, immune deviation to a Th2 response becomes more dominant. After severely dysplastic lesions become invasive, cervical cancer cells die and release HPV E6 and E7 oncoproteins that react with anti-E6 and anti-E7 antibodies to form insoluble immune complexes in antibody excess under the continuing influence of immune deviation. On the basis of this knowledge of the pathogenesis of advanced cervical cancer, we believe that successful immunotherapeutic treatments of these patients will use a vaccine formulation that will break peripheral tolerance in association with biological response modifiers that will enable the patient's immune system to switch classes from Th2 to Th1 while up-regulating MHC-1 molecules on cancer cells. Like prophylactic vaccines against HPV, successful therapeutic vaccine against cervical cancer may have to be universal rather than individualized to be efficacious. PMID- 11733944 TI - Decreased CD44v6 expression in lamina propria lymphocytes of patients with inflammatory bowel disease. AB - Splice variants of the glycoprotein CD44 are transiently expressed on lymphocytes during T cell activation. Increased expression of CD44v6 on peripheral blood lymphocytes (PBL) of patients with inflammatory bowel disease (IBD) was described recently. The aim of this study was therefore to characterize CD44v6 expression on CD4(+) lamina propria lymphocytes (LPL) of patients with active IBD in comparison to controls. CD44v6 expression on CD4(+) LPL (n = 19) of controls and patients with active IBD (Crohn's disease n = 14, ulcerative colitis n = 15) was analyzed by flow cytometry and compared to that on autologous PBL. Thereby, in vitro regulation of CD44v6 on LPL and PBL via CD3 and CD2 and the costimulatory signal B7-1 was examined. In addition, the role of protein kinase C (PKC) in CD44v6 expression was tested. CD44v6 expression was increased in CD4(+) LPL (median, 45%) compared to PBL (median, 38%). Surprisingly, in IBD CD44v6 was downregulated on CD4(+) lamina propria T cells, irrespective of their state of inflammation (median, 28%). CD44v6 expression on LPL was not upregulated upon CD3 activation alone but following costimulation with B7-1. However, CD2-mediated T cell activation sufficiently induced upregulation of CD44v6 on LPL and PBL. In our study, downregulation of CD44v6 on LPL of patients with IBD was not due to defective PKC activation. Taken together, these data indicate that decreased CD44v6 expression on LPL in IBD might be a feature of an inappropriate costimulatory signal in T cell activation. PMID- 11733945 TI - Hepatitis C viral proteins affect cell viability and membrane permeability. AB - To determine the effect of hepatitis C virus (HCV) proteins on cell growth, Huh-7 cells were transfected with a full-length HCV cDNA (pMO9.6-T7 Rz) clone and HCV proteins were expressed using a replication-defective adenovirus that encodes the gene for the T7 RNA polymerase. Expression of HCV proteins from this full-length clone resulted in reduction in viability of transfected cells as measured by trypan blue viability assay. For identification and separation of cells expressing hepatitis C virus proteins by fluorescence microscopy and flow cytometry, GFP was cloned in the HCV full-length clone. Cells transfected with the HCV-GFP chimera clone produced high levels of accurately processed structural and nonstructural proteins similar to those of the HCV full-length clone, which could be detected by Western blot analysis. Cells expressing all HCV proteins lost membrane permeability and underwent apoptotic cell death, indicated by the appearance of a sub-G0 peak in cell cycle analysis, DNA fragmentation in a TUNEL assay, and microscopic detection of nuclear condensation. Using double-channel flow analysis we confirmed that high-level expression of HCV proteins affected membrane permeability and cell survival. These results suggest that expression of all structural and nonstructural proteins from HCV cDNA in hepatic cells induces apoptotic cell death, which might be an important event in chronic hepatitis infection in humans. PMID- 11733946 TI - Deactivation of the innate cellular immune response following endotoxic and surgical injury. AB - The innate cellular immune (iCMI) system provides for the rapid production of interferon-gamma (IFN gamma) by NK cells in response to microbial threats. In this review, we examine the cellular and cytokine mechanisms of innate cellular immunity as determined in murine endotoxemia. This will be contrasted to the subsequent suppression of these same responses present in the mouse model of endotoxin tolerance, which is characterized by profound deficiency in both IL-12 and IFN gamma synthesis. Transient IFN gamma deficiency due to altered iCMI function has also been described in trauma or burn patients and is termed "clinical immune paralysis." If the common pathogenesis of these entities can be better understood, immune-based interventions might be identified for restoring iCMI function. In addition to the gain in basic immunologic insight, research on this subject may deliver future forms of prophylaxis against infection that do not rely on antibiotics and that will not promote antimicrobial resistance. PMID- 11733947 TI - READIT: a novel technology used in the interrogation of nucleic acid sequences for single-nucleotide polymorphisms. AB - The ability to perform analyses for single-nucleotide polymorphisms (SNPs) has become routine in many molecular diagnostic laboratories. While various procedures and technologies are available to do so, we evaluated a novel technology for SNP analysis using the Factor V Leiden polymorphism as an example. The Factor V Leiden polymorphism G1691A is the most common genetic abnormality associated with hereditary thrombophilia. Current testing methods remain highly accurate yet their main disadvantage is gel-based detection. The READIT System (Promega Corp., Madison, WI) is a novel approach to SNP analysis that utilizes DNA polymerase-mediated pyrophosphorolysis to release dNTPs, which are converted to ATP and used in a luciferase detection reaction. We screened 280 DNA specimens, previously analyzed using the PCR/MnlI restriction digest assay, to evaluate READIT System capabilities along with the KingFisher robotic magnetic particle processor (ThermoLabsystems). Concordant results were obtained for 278/280 (99%) specimens. One discordant result was due to an equivocal relative light unit while the other was indeterminate. Both specimens gave correct results upon repeat analysis. The READIT System offers several advantages including: (1) rapid SNP analysis, (2) accuracy and precision, (3) cost effectiveness, (4) decreased turn-around times, (5) high throughput, and (6) excellent analysis software. PMID- 11733948 TI - Activation of Kupffer cells during the course of carbon tetrachloride-induced liver injury and fibrosis in rats. AB - Kupffer cells are involved in the pathogenesis of chemically mediated liver injury through release of biologically active mediators that promote the pathogenic process. The purpose of this study was to elucidate specific biochemical and molecular changes occurring in Kupffer cells throughout a time course of carbon tetrachloride (CCl(4))-mediated liver injury and fibrosis. Rats were administered 1 ml/kg of CCl(4) (10% v/v olive oil) twice weekly for up to 6 weeks. Plasma alanine aminotransferase values and hematoxylin-and-eosin- and trichrome-stained liver sections indicated minor liver damage at 2 weeks followed by increased damage and collagen deposition by 4 and 6 weeks. Additionally, mRNA levels in Kupffer cells isolated from CCl(4)-treated rats demonstrated significant increases in tumor necrosis factor alpha (TNF alpha); tumor growth factor beta; interleukin-6 (IL-6); interleukin 1 beta; cyclooxygenase 2; CD14, and I kappa B alpha transcripts after 2 and 4 weeks of treatment. However, the expression of these genes at 6 weeks was similar to that of controls. Increased gene expression of cytokines in Kupffer cells isolated from CCl(4)-treated rats was accompanied by increases in protein production of TNF alpha, IL-6, IL-1 beta, and interleukin 10 following lipopolysaccharide stimulation. Further, liver sections stained for ED2-positive cells demonstrated an increase in the number of resident macrophages at 2 and 4 weeks with a slight decrease in ED2-positive cells by week 6 but still significantly more than control. Analysis of reduced glutathione (GSH) and oxidized glutathione (GSSG) indicated that Kupffer cells from CCl(4)-treated animals exhibited a 50% decrease in GSH at 2 and 4 weeks, whereas no significant changes were observed for GSSG. In conclusion, these data implicate Kupffer cells as a critical mediator of the inflammatory and fibrogenic responses during CCl(4)-mediated liver damage and provide new insight into the temporal molecular and biochemical changes associated with the ability of these resident macrophages to modulate liver injury. PMID- 11733949 TI - Aggresome formation in liver cells in response to different toxic mechanisms: role of the ubiquitin-proteasome pathway and the frameshift mutant of ubiquitin. AB - Aggresomes form in cells when intracellular proteins undergo conformational changes, as in so-called conformational diseases. This phenomenon has been observed in the liver and brain and in cell culture in response to abnormal protein formation, such as mutant proteins. In the case of the brain the frameshift mutant ubiquitin (UBB+1) is involved. Mallory body formation in the liver is one example of this phenomenon in vivo. Mallory body formation is common in a variety of liver diseases of diverse pathogenesis. The study of the Mallory body forming model indicated that drug-conditioned hepatocytes form Mallory bodies when mice are given colchicine, ethanol, okadaic acid, or exposure to heat shock. These findings suggest that aggresome formation is a common pathway of liver injury due to diverse mechanisms. To further characterize the role of this common pathway, drug-primed mice were exposed to different types of liver injury, i.e., using such drugs as thioacetamide, galactosamine, tautomycin, and the proteasome inhibitor PS341. Mallory body formation was induced by treatment with all the toxins tested, giving credence to the proposal that aggresome formation in the liver is a common pathway in response to different primary mechanisms of liver injury. The frameshift mutant UBB+1 was invariably found to colocalize with ubiquitin in the Mallory body, indicating its essential involvement in the mechanism of MB formation. PMID- 11733950 TI - Fibrosin: a novel lymphokine in wound healing. AB - Several growth factors are actively synthesized during wound repair and function to stimulate different cell types involved in the process of healing. Fibrosin is a novel fibrogenic lymphokine that stimulates several biological activities that relate to in vivo scarring. To investigate the role of fibrosin, we used "punch biopsy" and linear wounding procedures in a murine model of wound healing. Histological examination showed that recombinant fibrosin stimulated epithelialization of wounds and accelerated healing of both punch biopsy and linear wounds. Fibrosin enhanced healing of linear wounds by reducing the time for healing by approximately 30-40%. From our data we estimated the healing time of control wounds to be 22-24 days; wounds treated with fibrosin appeared to heal in 14-16 days. Our observations suggest that fibrosin enhances wound healing and may be involved in accelerating epithelialization, collagen matrix formation, and also remodeling of the extracellular matrix in vivo. Thus fibrosin may function during different phases of wound healing and act as a potent inducer of scar formation and wound healing. This finding may have direct clinical applications. PMID- 11733951 TI - Computer-assisted image analysis of the rat postosteonecrotic remodeled femoral head. AB - Osteonecrosis of rat femoral heads was induced by stripping the periosteum of the neck and cutting the ligamentum teres. The epiphyseal marrow and bone were necrotic on the 5th postoperative day. Specimens obtained 18 and 36 days postoperatively showed fibrous and hematopoietic-fatty tissue in the intertrabecular spaces, osteoclastic bone resorption, osteogenesis, and degeneration of the joint cartilage. Morphometrically, the means of the height-to length ratios of the control, 6-day, 18-day, and 36-day femoral heads were 0.26, 0.28, 0.48, and 0.29, respectively. The shape factor of the femoral heads of the control rats was higher than 0.81 in 80% of the cases, while those of rats killed on the 6th, 18th, and 36th postoperative day were higher than 0.81 in 65, 60, and 50% of cases, respectively. Statistically, the means of the height-to-length ratios and the values of the shape factors of the femoral heads of the rats killed 18 days postoperatively differed significantly from those of the other three groups of rats. The quantitatively gauged data of the remodeled epiphyses negate the authors' subjective impression concerning early flattening of the femoral heads after surgically produced osteonecrosis. PMID- 11733952 TI - Functional and compositional studies of arteries stored in University of Wisconsin solution compared with Krebs-Bulbring buffer. AB - Endothelium-dependent and -independent vasorelaxation in ring segments of rabbit thoracic aorta is reduced and noradrenaline-induced vasoconstriction unaltered after prolonged storage in University of Wisconsin solution (UW) compared to arteries stored in extracellular-type solutions such as Krebs-Bulbring buffer (KBB). The aims of the present study were to determine whether angiotensin-II induced vasoconstriction, alterations in myosin light chains, protein synthetic capacity, and subcellular structures are altered after 8 days of UW storage at 4 degrees C. The present study showed reduced contractility to angiotensin II, following 8 days of cold storage in UW, that was not reversed in the presence of a nitric oxide synthase inhibitor, N(G)-nitro-l-arginine methyl ester (100 microM). Measurements of contractile protein ratios in the same tissues after cold storage in UW or KBB did not show any significant alterations in smooth muscle myosin light chains or protein synthetic capacity (reflected by total RNA). It is concluded that reductions in vasoconstriction in UW-stored tissue are unlikely to be due to increased release of nitric oxide nor reduced availability of myosin light chains for phosphorylation and vasoconstriction. PMID- 11733953 TI - Why study third-, fourth-, fifth-, ...line chemotherapy of ovarian cancer? PMID- 11733954 TI - Is there any progress in the outcome of patients suffering from ovarian cancer? Treatment strategies since 1957. PMID- 11733955 TI - Vulvar melanoma: a retrospective analysis and literature review. AB - OBJECTIVE: This review focuses on current directions in the staging and treatment of melanoma of the vulva. METHODS: All women treated for invasive melanoma of the vulva at the University of Virginia Health Sciences Center from 1980 through 2000 were identified through a retrospective review of the records of the Division of Gynecologic Oncology. Their treatments and outcomes were then analyzed and presented. RESULTS: Over the 20-year study period, 14 cases of melanoma of the vulva were identified. Of the 14 patients treated with curative intent, 6 developed recurrences following the completion of primary therapy, and all are dead from their disease. The mean duration from completion of therapy to recurrence was 7.5 months; the mean survival following recurrence was 17 months. CONCLUSION: One-centimeter skin margins appear adequate for vulvar melanomas <1 mm thick, and 2-cm margins appear adequate for intermediate-thickness melanomas (1-4 mm). In all cases it is necessary to include at least a 1-cm-deep margin extending through the subcutaneous fat to the muscular fascia below. Elective node dissection seems to offer no additional advantage in superficial lesions <0.76 mm thick, and its role in deeper lesions is still uncertain. PMID- 11733956 TI - Types and duration of symptoms prior to diagnosis of invasive or borderline ovarian tumor. AB - OBJECTIVE: The objective was to describe and compare types and duration of symptoms among women with invasive versus borderline ovarian tumors. METHODS: Cases were women, ages 20-69 years, diagnosed with invasive (616) and borderline (151) epithelial ovarian tumors from 1994 to 1998. Symptoms were obtained using a standardized in-person interview. Differences in types and duration of symptoms, time to diagnosis after consulting a physician, and primary reason for diagnosis by invasive/borderline status and histologic type were determined using bivariate and regression analyses controlling for age. RESULTS: Borderline and invasive cases reported similar types of symptoms. However, borderline cases were twice as likely to report not having had symptoms as invasive cases (16 vs 8%, P = 0.005). Prediagnostic symptom duration was longer among borderline versus invasive cases (median: 6 vs 4 months, P < 0.001). The median time from first consultation with a physician to diagnosis (1 month) did not differ by invasive/borderline status. Borderline cases were twice as likely to be diagnosed through routine examination as invasive cases (28 vs 16%, P = 0.001). Invasive cases were more likely to be diagnosed because of symptoms (62 vs 48%, P = 0.002). CONCLUSIONS: Because most (90%) women with ovarian tumors have symptoms and median symptom duration is 4 months, greater awareness of symptoms by women and physicians is needed for the earlier detection of ovarian tumors. The lesser likelihood of being detected by routine examination and the shorter symptom duration for invasive versus borderline cases underscores the need for effective screening and preventive strategies. PMID- 11733957 TI - Pulmonary resection in metastatic uterine and cervical malignancies. AB - OBJECTIVE: Although thoracotomy for removal of pulmonary metastasis is well documented in a wide variety of solid tumors, data are sparse regarding management of patients with gynecologic malignancies metastatic to the lung. METHODS: We retrospectively reviewed the Roswell Park Cancer Institute experience between 1982 and 1999. Of 82 eligible patients with gynecologic tumors metastatic and confined to the lung, 25 underwent pulmonary resection. RESULTS: There were 60 uterine and 22 cervix cancer patients with pulmonary metastases. Among patients with uterine cancer primaries undergoing pulmonary resection (n = 19) median survival was 26 months. Uterine cancer patients who underwent surgical resection for leiomyosarcomas (n = 11) had a median survival of 25 months compared to 46 months in patients with adenocarcinoma (n = 6, P = 0.02). Median survival in cervix cancer patients undergoing resection for pulmonary metastases (n = 6) was 36 months. CONCLUSIONS: Pulmonary resection may provide a survival advantage for selected patients with uterine and cervical malignancies with metastases isolated to the lung. PMID- 11733958 TI - A phase I/II study of topotecan in combination with carboplatin in recurrent epithelial ovarian cancer. AB - OBJECTIVE: The purpose of this study was to define the maximum tolerated dose (MTD) of topotecan given as escalating doses combined to a fixed dosage of carboplatin in late relapsing ovarian carcinomas. METHODS: Women with relapsing ovarian cancer more than 6 months after first-line treatment were eligible for the study. In the first phase of the trial, patients were allocated to escalating topotecan doses with a carboplatin fixed dose (AUC 5, according to Cockcroft's formula). If no "severe" adverse event occurred in 1 or more of the patients, the topotecan dose was increased. The starting dose of topotecan was 0.50 mg/m(2)/day, for 3 consecutive days, and the dose step was of 0.25 mg/m(2)/day, till 1.5 mg/m(2)/day. The study progressed then in a phase II trial. RESULTS: A total of 39 patients entered the trial. Twenty took part in the escalating topotecan dose phase (4 per dose level, 0.50, 0.75, 1, 1.25, and 1.50 mg/m(2)/day) and 19 in the phase II. No severe adverse event was observed in the phase I of the trial, so the MTD was not reached. In the phase II trial topotecan was given to 1 mg/m(2)/day. Overall grade 3-4 neutropenia, lasting 7 days or less, was observed in 58.9% (23 patients). Thrombocytopenia occurred in 30.8% (12 patients) and grade 3 anemia in 25.6% (10 patients) of subjects. No life threatening event occurred. Platelets or red blood cell transfusions were given in three cases (7.8%). CONCLUSIONS: This daily-times-3-day schedule of topotecan in combination with carboplatin is safe. PMID- 11733959 TI - Is the decision for colorectal resection justified by histopathologic findings: a prospective study of 100 patients with advanced ovarian cancer. AB - OBJECTIVE: We compare the indication for colorectal resection in patients with advanced ovarian cancer with histopathologic findings. We describe the effect on pelvic control and morbidity associated with surgery. METHODS: Between February 1995 and March 2001, 100 patients with FIGO stage IIIc ovarian cancer underwent pelvic en bloc resection with excision of the rectosigmoid colon as part of primary or secondary cytoreductive surgery. Decision for resection was made by the surgeon when tumor involvement of the cul-de-sac was suspected. Rectosigmoid infiltration was histopathologically defined as infiltration of the serosa or deeper. RESULTS: In 73 of 100 patients (73%) tumor involvement of the rectum was confirmed histopathologically: infiltration of the serosa in 28 (28%) patients, infiltration of the muscularis in 31 (31%) patients, and infiltration of the mucosa in 14 (14%) patients; in 27 (27%) patients no infiltration was found. Histopathologically confirmed pelvic R0 resection was achieved in 85 (85%) patients. In 11 (11%) patients the pelvic resection margins were tumor-involved and in four (4%) patients visible parametric tumor remained in situ. Pelvic recurrence occurred in 4 (4.7%) of 85 optimally debulked patients compared with 9 (60%) of 15 patients with suboptimal pelvic resection status (P < 0.05). End colostomy could be prevented in 94 (94%) of 100 patients. CONCLUSION: Pelvic en bloc surgery with rectosigmoid resection was justified by histopathologic outcome since deperitonealization with preservation of the rectosigmoid would have left tumor in situ in 73% of patients with suspected cul-de-sac involvement. PMID- 11733960 TI - Dominant-negative mutation of p53 tumor suppressor gene in endometrial carcinoma. AB - BACKGROUND: It has been suggested that mutation of the TP53 tumor suppressor gene is involved in endometrial carcinogenesis. However, the status of p53 function in endometrial cancers has not yet been investigated in detail. METHODS: We surveyed inactivating p53 mutations in endometrial carcinomas using the yeast p53 functional assay, which can evaluate the transcriptional activity of p53 in vivo in yeast. To the detected p53 mutants, we also applied a transdominance assay, which assesses the dominant-negative property of mutants. RESULTS: Of 23 endometrial carcinomas, 9 tumors (39.1%) were found to harbor p53 mutations. Only 1 of the 6 mutants in 18 endometrioid-type tumors showed dominant-negative capacity. In contrast to the endometrioid-type tumor, all 3 mutations in 5 serous type tumors (R273H, 9-bp deletion in codons 240-243, and R248W) showed dominant negative capacity and presented in a homozygous state in the tumors, indicating a complete functional inactivation. CONCLUSIONS: Although this study included a relatively small number of cases and therefore is a preliminary study, these results suggest that the dominant-negative mutation of the TP53 gene is related to serous adenocarcinoma. The role of the dominant-negative status of p53 mutants in endometrial carcinogenesis and progression of this disease should be further investigated. PMID- 11733961 TI - Loss of expression of the p16 tumor suppressor gene is more frequent in advanced ovarian cancers lacking p53 mutations. AB - OBJECTIVE: The aim of this study was to test the hypothesis that p53 mutations are less frequent in ovarian cancers with alterations in other genes that regulate G1 progression. METHODS: Expression of G1 stimulatory (cyclins D1 and E, cdk4, Ki67) and inhibitory (p16, Rb, p27, p14) genes was analyzed using Western blots in 84 primary ovarian cancers and seven cell lines of known p53 mutation status. Expression of p16 and Rb also was determined using immunohistochemistry and the p16 gene was examined for homozygous deletions and mutations. RESULTS: Loss of p16 protein was more frequent in ovarian cancers with wild-type p53. All four cell lines with wild-type p53 had lost p16 compared to only one of three with mutant p53 genes. p16 expression was absent in 34% (28/82) of primary ovarian cancers, and this was significantly more common in cases with wild-type p53 (14/28, 50%) compared to those with p53 mutations (14/54, 26%, P = 0.03). Homozygous deletion of the p16 gene was found in cell lines lacking p16, but not in any primary cancers. p16 loss was more common in serous (21/52, 40%) than nonserous cancers (4/23, 17%, P = 0.07). Cases that expressed p16 were more likely to express high levels of Rb (47/55, 85%) than p16-negative cases (12/28, 43%, P < 0.001). Loss of Rb occurred in 5/30 (17%) ovarian cancers lacking p53 mutations compared to 5/54 (9%) cases with p53 mutations (P = 0.48). Expression of G1 stimulatory proteins (cyclins D1 and E, cdk4, Ki67) did not correlate with p53 mutation status. CONCLUSIONS: Loss of expression of the p16 tumor suppressor occurs more often in ovarian cancers lacking p53 mutations. These data are consistent with the paradigm that inactivation of p53 is less of a requisite event in ovarian carcinogenesis when another G1 regulatory gene such as p16 already has been inactivated. PMID- 11733962 TI - Papillary serous carcinoma of the uterus: increased risk of subsequent or concurrent development of breast carcinoma. AB - OBJECTIVE: Some women with endometrial cancer may be at increased risk for developing breast cancer. The histologic type of endometrial cancer associated with synchronous or subsequent breast cancer has not been clearly established. Our purpose was to determine if a certain histologic type of endometrial cancer was associated with an increased risk of synchronous or subsequent breast cancer. METHODS: The University of Iowa Hospitals and Clinics tumor registry was queried to ascertain all patients with the diagnosis of uterine cancer from January 1, 1983, to December 31, 1994. Statistics were performed utilizing SPSS for Windows version 9.0 (SPSS Inc., Chicago, IL), including Student's t tests and chi(2) tests. RESULTS: Five hundred ninety-two patients had endometrial adenocarcinoma during the study period. Five hundred thirty-six women had endometrioid adenocarcinoma, 23 women had papillary serous carcinoma (UPSC), 21 women had adenosquamous carcinoma, 10 women had clear-cell carcinoma, and 1 woman each had mucinous or squamous carcinoma. Twelve patients had previously been diagnosed with breast carcinomas. Twenty-five patients were diagnosed with breast cancer either concurrently or subsequent to their diagnosis of endometrial cancer. Synchronous or subsequent breast cancers developed in 3.2% of patients with endometrioid carcinoma and in 25% of patients with UPSC (P < 0.001). CONCLUSION: Patients with UPSC have an increased risk of development of breast cancer as compared to patients with endometrioid adenocarcinoma of the uterus. PMID- 11733963 TI - Secondary cytoreductive surgery for patients with recurrent epithelial ovarian carcinoma. AB - OBJECTIVE: The aim of this prospective study was to evaluate the influence of secondary cytoreductive surgery on survival of patients with recurrent epithelial ovarian cancer. METHODS: Between June 1993 and June 1999, 149 patients after primary treatment underwent secondary cytoreductive surgery: 69 (46.3%) had recurrence-free interval (RFI) 7-12 months, 59 (39.6%) RFI 13-24 months, and 21 (14.1%) RFI >24 months. Exclusion criteria included secondary cytoreduction during second-look laparotomy, interval cytoreduction, and palliative surgery in patients with intestinal obstruction or progressive disease. RESULTS: The median follow-up was 27 months. According to multivariate analysis, RFI groups, prior chemotherapy combination, and residual tumor after secondary surgery were associated independently with overall survival. Residual tumor after secondary surgery was by far the most strongly predictive factor for patient's survival (hazard ratio (HR) 2.65; 95% confidence interval (CI) 1.43-4.92). The 2-year survival rates were 22.3, 62.9, and 22.7%, respectively, for patients with RFI 7 12, 13-24, and >24 months. The 5-year survival was 29% for patients with RFI 13 24 months. No patients with RFI 7-12 and >24 months outlived the 4-year estimate. Unexpectedly, RFI >24 months was not correlated significantly with overall survival. However, 17 patients (81%) with RFI >24 months were heavily treated with chemotherapy before secondary surgery. CONCLUSIONS: Patients who have documented gross disease preoperatively should be selected for a secondary debulking operation; the entity of surgical effort would be modulated in relation to previous treatment. PMID- 11733964 TI - Outcome and management of pathological stage I endometrial carcinoma patients with involvement of the lower uterine segment. AB - OBJECTIVE: The objective was to evaluate the clinicopathologic characteristics and outcome of pathologic stage I endometrial carcinoma patients with lower uterine segment (LUS) involvement. METHODS: We retrospectively reviewed the characteristics and outcomes of pathologic stage I endometrial carcinoma patients treated with primary surgery at our institution between 1988 and 1998. The significance of LUS involvement was examined with univariate and multivariate analyses. Median patient follow-up was 37.3 months. RESULTS: Of the 98 cases reviewed, 41 (42%) had LUS involvement. No differences were seen in the clinicopathologic features, extent of surgical staging, or adjuvant therapies between patients with and without LUS involvement. Univariate analysis revealed that grade, lymphovascular invasion (LVI), myometrial invasion (MI), and histology were correlated with recurrence. While the 5-year actuarial disease free survival was worse in women with LUS involvement (80.3 vs 94.0%) compared to those without, this difference did not reach statistical significance (P = 0.14). Moreover, after controlling for pathologic features in a multivariate model, LUS involvement was not correlated with patient outcome (P = 0.98; hazard rate 0.97; 95% confidence interval 0.24, 4.0). LUS was also not correlated with pelvic recurrence. Of 25 low-risk patients (superficial MI and grade 1-2 disease) with LUS involvement, none recurred in the pelvis following surgery alone. In contrast, pelvic recurrence was common (5/12 or 41.6%) in high-risk patients (deep MI and/or grade 3 tumors) following surgery alone regardless of LUS involvement. CONCLUSION: LUS involvement is common in pathologic stage I endometrial carcinoma but is not correlated with a worse outcome. Moreover, in the absence of adverse pathologic features, LUS involvement is not associated with an increased risk of pelvic recurrence and should not be used as an indication for adjuvant radiation therapy. PMID- 11733965 TI - Panniculectomy with simultaneous gynecologic oncology surgery. AB - OBJECTIVE: The objective was to report the utility and morbidity of panniculectomy in obese gynecologic oncology patients undergoing exploratory laparotomy. METHODS: A retrospective chart review of 41 consecutive women who had a panniculectomy as part of an abdominal gynecologic oncology procedure between July 1996 and May 2000 was performed. Obese patients possessing a large pannus, the majority with a BMI > or = 30 kg/m(2), were included. Demographic, preoperative, operative, and postoperative data were obtained. Statistical analyses were performed using Statistical Analysis System (SAS) Version 6.13. RESULTS: Panniculectomy was performed on 41 patients with a mean age of 55, weight of 126 kg, and BMI of 48 kg/m(2). The most common comorbidities in this population were hypertension, diabetes, and osteoarthritis. Wound infection occurred in 4 (9.8%) patients; 88% of the patients received a hysterectomy. The average EBL was 358 cc. Operative time and length of hospital stay were on average 203 min and 5.5 days, respectively. A prior history of diabetes increased the risk of early complications (P = 0.03). Late complications were more likely to occur in older women (P = 0.05). Wound complications were increased in patients with larger BMI's (P = 0.05). CONCLUSIONS: This study supports the safety of the panniculectomy procedure in this high-risk group of morbidly obese patients for whom a technical advantage may be achieved by improved operative exposure. PMID- 11733966 TI - Characterization of a human carcinosarcoma cell line of the ovary established after in vivo change of histologic differentiation. AB - OBJECTIVES: Cell lines are valuable in vitro models for clinical and basic research. Most ovarian cancer cell lines described are serous cystadenocarcinomas or poorly differentiated adenocarcinomas. The establishment of ovarian cancer cell lines with rare histologic differentiation is especially of interest. We describe the establishment of a carcinosarcoma cell line of the ovary after in vivo selection. METHODS: The cell line OV-MZ-22 was established from a solid tumor mass in the upper abdomen. At the time of establishment, the patient underwent secondary debulking and was pretreated with six cycles of cis platinum/epirubicin/cyclophosphamide. Features of the cell line studied included morphology, ultrastructure, heterotransplantation, chromosome analysis, and analysis of intermediate filament proteins and actins by immunocytochemistry. RESULTS: The first histologic report of the patient described a papillary cystadenocarcinoma, which changed to a carcinosarcoma with predominantly sarcomatous differentiation at secondary debulking. This cell line is aneuploid and shows no expression of the tumor-associated antigens CA-125 and CEA, but an overexpression of MDR-1, lung resistance protein, p53, and topoisomerase I and II, but not of multidrug-resistance-associated protein. The cell line did not give rise to transplant tumors in nude mice. The histologic and immunocytochemical comparison of the primary and the relapsed tumor proved evidence of an in vivo change of differentiation from predominantly papillary cystadenocarcinoma to carcinosarcoma. Morphological characteristics and intermediate filament pattern underlined the sarcomatous differentiation and origin of this cell line. The differentiation phenotype of OV-MZ-22 cells is that of smooth-muscle cells. CONCLUSION: The change of histologic differentiation was apparently due to a selection process caused by platinum-containing chemotherapy. The origin of the cell line and its rarity make this new line an appropriate tool for further investigation. PMID- 11733967 TI - Factors predicting subcutaneous implanted central venous port function: the relationship between catheter tip location and port failure in patients with gynecologic malignancies. AB - OBJECTIVE: We set out to determine the factors that predict subcutaneous implanted central venous port function. Specifically, we sought to determine whether the location of the catheter tip is correlated with port failure. METHODS: A review of all gynecologic oncology patients who underwent initial port placement between 1993 and 1998 was undertaken. The initial chest radiograph following port placement was reviewed, and the venous location of the catheter tip was recorded. Patients were followed until port removal, death, or the last documentation of port function. RESULTS: Two hundred thirty-six patients underwent port placement during the study period. The majority of patients (97%) had their port placed for intravenous chemotherapy. The median time of port duration in patients with a functional port was 21.6 months. Forty of the 236 ports (17%) were removed because of device malfunction. Catheter tips were located in the central venous system in 164 (69%) cases and outside of the central venous system in 72 (31%) cases. Removal secondary to malfunction was significantly higher when the catheter tip was located outside of the central venous system (30/72 (42%) versus 10/164 (6%), P = 0.001). By life-table analysis, ports removed for malfunction with their tips located centrally had a significantly longer median duration of functional use than those whose tips were located peripherally (78 versus 44 months, P = 0.0001). CONCLUSIONS: The rate of port removal secondary to malfunction is significantly less if the catheter tip is located in the central venous system. Confirmation of the location of the catheter tip is imperative for the long-term function of a subcutaneous implanted central venous port. PMID- 11733968 TI - "What doesn't kill you makes you stronger": an ovarian cancer survivor survey. AB - OBJECTIVES: This survey was designed to learn more about the physical health and quality of life of ovarian cancer survivors without known active disease and not on treatment. METHODS: Women with ovarian cancer without evidence of active disease, and not on treatment for at least 2 years, were recruited in Canada and the United States from university cancer clinics, community ovarian support groups, and a cancer survivor network newsletter and asked to complete an anonymous mail-back survey on their physical, psychological, social, and spiritual well-being. RESULTS: Two-hundred (67.5%) women responded. Their mean age was 55.3 years, 72.5% had children, and they had been diagnosed with ovarian cancer a mean of 7.2 years previously. Eighty-nine percent regarded their health as good or excellent, but 53.5% had current pain or discomfort. They reported better mental health and equivalent energy levels to the general population. Although 57% reported that their sex lives had been negatively affected by cancer and its treatment, their overall sense of loss relating to sexual functioning was moderate to low. However, women under age 55 years reported a greater sense of loss about sexual function and fertility (P = 0.001). Most women reported that their ovarian cancer experience had changed their views on life and relationships in an overwhelmingly positive way. CONCLUSION: Most ovarian cancer survivors, despite some symptoms, enjoyed good physical, psychological, social, and spiritual health. Specific concerns emerged about sexuality which should be addressed in patient education. Having survived a life-threatening illness, ovarian cancer survivors appear to have put other life difficulties into perspective, altered their priorities, and felt enriched by the experience. In general, these women showed impressive and inspiring resilience and reported good physical health and energy, excellent psychological health, and feelings of greater pleasure in life and personal relationships. PMID- 11733969 TI - Expression of cell-cycle mediators in ovarian cancer cells after transfection with p16(INK4a), p21(WAF1/Cip-1), and p53. AB - OBJECTIVE: The purpose of this study was to determine whether transfection of ovarian cancer cell lines with recombinant adenoviral vectors containing wild type p16(INK4a), p21(WAF1/Cip-1), and p53 caused growth inhibition and induction of apoptosis. We also measured the expression of the cell-cycle mediators Bax, Bcl-2, pRb, and mdm-2. METHODS: We introduced the wild-type p16(INK4a), p21(WAF1/Cip-1), and p53 genes into the ovarian cancer cell lines SK-OV-3 (p16(INK4a) and p53 null) and OVCA-420 (p16(INK4a) and p53 wild-type) by adenoviral transfection. Cell growth inhibition was measured over a 10-day period. Induction of apoptosis was tested for both cell lines 48 h after cell transfection. Expression of cell-cycle mediators was evaluated by Western blot analysis and densitometry. RESULTS: Growth inhibition was documented after transfection with p16(INK4a), p21(WAF1/Cip-1), and p53 in both SK-OV-3 cells and OVCA-420 cells. Apoptosis was greatest in SKOV-3 cells after transfection with p53. A significant expression of Bax was only seen in the SKOV-3 cells transfected with p53. The bcl-2 protein was poorly expressed in both cell lines. Expression of pRb was suppressed in OVCA-420 cells transfected with p16(INK4a) and p21(WAF1/Cip-1). Infection with Adp16(INK4a) and Adp53 led to an increase in the level of mdm-2 in the SK-OV-3 cell line only. CONCLUSIONS: In the ovarian cancer cell lines studied, cell growth was inhibited after transfection with p16(INK4a), p21(WAF1/Cip-1), and p53. Cell cycle arrest was highest with p53 transfection. The expression of pro-apoptosis proteins was primarily a function of p53 expression. PMID- 11733970 TI - Adenovirus-mediated thymidine kinase gene therapy in combination with topotecan for patients with recurrent ovarian cancer: 2.5-year follow-up. AB - OBJECTIVE: Patients with recurrent ovarian cancer were treated intraperitoneally (ip) with a replication-deficient recombinant adenovirus (ADV) containing the herpes simplex virus thymidine kinase gene. Vector delivery was followed by intravenous administration of an antiherpetic prodrug and a topoisomerase I inhibitor. METHODS: Ten patients with stage IIIc epithelial ovarian cancer underwent secondary debulking to < or =0.5 cm residual tumor. Patients with normal ip flow received delivery of ip ADV. Two patients each were treated on dose level 1 (2 x 10(10) vector particles), dose level 2 (2 x 10(11)), and dose level 3 (2 x 10(12)); four patients were on dose level 4 (2 x 10(13)). Acyclovir and topotecan were started 24 h after vector delivery. Five patients underwent second-look surgery about 4 weeks after application of gene therapy (GT). RESULTS: At the time of the second-look surgery, two out of five patients were free of tumor. Four weeks after GT none of the peritoneal biopsies showed residual vector DNA. Patients pretreated with an average of three different chemotherapeutic drugs and two different chemotherapy regimens had a median overall survival (OS) of 18.5 months. Three patients are still alive 30, 30, and 31 months after GT. CONCLUSION: With the combination of secondary optimal debulking, GT, and topotecan, median OS was about one-third longer than in previously reported second-and third-line trials. Survival was comparable to that of patients of other studies with secondary cytoreductive surgery in combination with chemotherapy. PMID- 11733971 TI - Mitotic index and ki-67 nuclear antigen labeling index as predictors of chemotherapy response in uterine cervical carcinoma. AB - OBJECTIVE: The aim of this study was to determine if the mitotic index (MI) and the Ki-67 nuclear antigen labeling index (Ki67LI) obtained from biopsy specimens could be used as predictors of chemotherapy response in uterine cervical carcinomas. METHODS: Six patients with squamous cell carcinoma who received neoadjuvant chemotherapy before radical surgery were entered into the study. The MI and the Ki67LI were evaluated using hematoxylin and eosin (H&E)-stained and immunostained slides before and after chemotherapy. Tumor size was measured three dimensionally by magnetic resonance imaging. We compared the values of MI and Ki67LI with changes in tumor size. RESULTS: The cases were classified according to the percentage reduction in tumor mass: one case showed a marked response (98%), four showed a moderate response (53-78%), and one showed a slight response (approximately 0%). In the case with a marked response, the MI values before chemotherapy and 3 and 7 days after chemotherapy were 15, 2, and 0, respectively. The corresponding Ki67LI values were 51, 16, and 0, respectively. In the moderate response cases, the corresponding MI values before and 3 and 7 days after chemotherapy were 3.9-13.6, 0.8-1.6, and 1.6-4.0, respectively. The Ki67LI was 21.8-44.2 before chemotherapy, with two cases increasing and two decreasing at 7 days after chemotherapy. In the case showing a slight response, the MI values before chemotherapy and 3 and 7 days after chemotherapy were 0.5, 0.8, and 1.0, respectively. The Ki67LI was 4.0 before chemotherapy and increased slightly to 6.0 at 7 days after chemotherapy. CONCLUSIONS: In six cases examined, high MI and Ki67LI values before chemotherapy and a marked decrease in MI shortly after chemotherapy appeared to be predictors of good neoadjuvant chemotherapy response in uterine cervical carcinomas. PMID- 11733972 TI - Detection of Mycoplasma ribosomal DNA sequences in ovarian tumors by nested PCR. AB - OBJECTIVE: The aim of this study was to investigate the reported association between mycoplasma infection and ovarian cancer by screening ovarian tumor tissues for the presence of mycoplasma DNA. METHODS: Forty-six benign and malignant ovarian tumors were obtained from patients undergoing pelvic surgery at a regional cancer center. DNA was isolated from snap-frozen tumor tissues, and commercial nested polymerase chain reaction (PCR) kits were used to detect the presence of 12 species of mycoplasma in tumor DNA samples. PCR products were isolated from ethidium bromide-stained agarose gels, and sequenced with an automated DNA sequencer. Species were identified through nucleotide sequence similarity searches using the National Center for Biotechnology Information BLAST program. RESULTS: Mycoplasma DNA was detected in 6 (13.0%) of the 46 tumor DNA samples. Nucleotide sequence similarity searches of nested PCR products revealed that one Mycoplasma salivarium and five M. arginini DNA sequences were amplified from the ovarian tissues. CONCLUSIONS: Since M. salivarium and M. arginini are frequently encountered laboratory contaminants that do not have a recognized role as human pathogens, our findings do not support an association between human mycoplasma pathogens and ovarian cancer. PMID- 11733973 TI - Surgical management of early-stage endometrial cancer in the elderly: is laparoscopy feasible? AB - OBJECTIVE: To give insight into the utility of laparoscopic staging of endometrial cancer in the elderly population by reviewing the surgical management of clinically stage I endometrial cancer patients. METHODS: A retrospective analysis evaluating patients that were > or =65 years old and had planned laparoscopic staging, traditional staging via a laparotomy, or a transvaginal hysterectomy as management of their early endometrial cancer. The laparoscopic group had complete staging with bilateral pelvic and paraaortic lymph node dissections and was compared to the group who had staging performed via laparotomy. Patients were identified by our institution's database and data were collected by review of their medical records. Data were collected on demographics, pathology, and procedural information including completion rates, operating room (OR) time, estimated blood loss (EBL), transfusions, lymph node count, complications, and length of stay. Associations between variables were analyzed by Student's t tests and chi(2) testing using Excel v. 9.0. RESULTS: From February 25, 1994, through December 21, 2000, 125 elderly patients were identified. Sixty-seven patients had planned laparoscopic staging (Group 1), 45 patients had staging via planned laparotomy (Group 2), and 13 patients had a transvaginal hysterectomy (Group 3). Group 1 and Group 2 were compared regarding surgical and postoperative data. Age was not different between these groups (75.9 vs 74.7 years, P = NS). Quetelet index was also similar (29.4 vs 29.9, P = NS) 32.8% of Group 1 had > or =1 previous laparotomy compared to 51.1% in Group 2 (P = NS). In Group 1, 53/67 (79.1%) had stage I or II disease compared to 29/45 (64.4%) in Group 2 (P = NS). Laparoscopy was completed in 52/67 (77.6%) attempted procedures. The reasons for conversion to laparotomy were obesity 7/67 (10.4%), bleeding 4/67 (6.0%), intraperitoneal cancer 3/67 (4.5%), and adhesions 1/67 (1.5%). OR time was significantly longer in successful Group 1 patients compared to Group 2 patients (236 vs 148 min, p = 0.0001). EBL was similar between these groups (298 vs 336 ml, P = NS). Ten of 52 (19.2%) of successful Group 1 patients received a blood transfusion compared to 1/45 (2.2%) of Group 2 patients (P < 0.0001). Pelvic, common iliac, and paraaortic lymph node counts were similar between successful Group 1 patients and those in Group 2 combined with those that received a laparotomy in Group 1 (17.8, 5.2, 6.6 vs 19.1, 5.1, 5.2, P = NS). Length of stay (LOS) was significantly shorter in Group 1 versus Group 2 (3.0 vs 5.8 days, P < 0.0001). There were less fevers (6.0 vs 15.6%, P = 0.01), less postoperative ileus's (0 vs 15.6%, P < 0.001), and less wound complications (6.0 vs 26.7%, P = 0.002) in Group 1 compared to Group 2. Group 3 average age was 77.5 years. Concurrent medical comorbidities were the main reason for the transvaginal approach. OR time averaged 104.5 min. The average length of stay was 2.1 days with no procedural or postoperative complications. CONCLUSIONS: The favorable results from this retrospective study refute the bias that age is a relative contraindication to laparoscopic surgery. Laparoscopic staging was associated with an increased OR time and an increased rate of transfusion but equivalent blood loss and lymph node counts. Possible advantages are decreased length of stay, less postoperative ileus, and less infections complications. Transvaginal hysterectomy still remains a proven option for women with serious comorbid medical problems with short OR times, minimal complications, and short lengths of stay. PMID- 11733974 TI - Altered CD44 variant 6 expression in FIGO stage IB cervical carcinoma. AB - OBJECTIVE: CD44 is an adhesion molecule which plays an important role in metastatic cascade by mediating tumor cell interaction with the endothelium and the subendothelial matrix. In this study CD44v6 expression was immunohistochemically investigated on 88 uterine cervical cancers. Correlation between expression and prognostic variables and the survival was examined. METHODS: Eighty-eight patients with stage IB disease, treated primarily with surgery, were examined histopathologically and immunohistochemically. CD44v6 expressions of tumoral tissue and the nonneoplastic tissue nearby were examined using antiCD44v6 monoclonal antibody. CD44v6 expression was compared to the known clinicopathologic prognostic variables and survival of patients. RESULTS: Nonneoplastic epithelium of the sections showed CD44v6 expression predominantly in basal and parabasal layers at least in traces. CD44v6 overexpression in neoplastic islands was evaluated as "general," "basal" (only in the basal portion of neoplastic islands), and "nonbasal" (also in the central portion of the neoplastic islands) separately. When expression and prognostic variables were compared, CD44v6 non-basal expression was found to be significant in nonsquamous cancers, when the tumor diameter was greater than 3 cm and in the tumors that showed recurrences. Univariate survival analysis with the Kaplan-Meier method showed that only the age of the patient is significantly correlated with disease free survival. Interestingly when the same analysis was done for 5-year overall survival, diameter of the primary tumor, depth of cervical stromal invasion, existence and number of lymph node involvement, positivity for general CD44v6 expression, and positivity for nonbasal expression were found to be statistically significant. Furthermore multivariate analysis with Cox regression showed that nonbasal CD44v6 expression and lymph node involvement are independent variables for 5-year overall survival. CONCLUSION: These results indicate that CD44v6 expression is associated with some of the important clinicopathologic prognostic variables and appears to be a predictor of advanced pathological-surgical stage of early clinical stage cervical carcinoma. CD44v6 nonbasal expression is significantly correlated with overall survival. PMID- 11733975 TI - Risk factors for epithelial borderline ovarian tumors: results of a Swedish case control study. AB - OBJECTIVE: Borderline ovarian tumors have a favorable prognosis. Previous epidemiological studies indicate common risk factors for invasive epithelial ovarian cancers and borderline tumors, but it remains unresolved whether these tumors are precursors of invasive cancers or a separate disease entity. The objective of this population-based case-control study conducted in 1993-1995 was to examine reproductive and other factors in relation to the risk of borderline ovarian tumors. METHODS: Subjects were 193 histologically verified incident epithelial borderline tumor cases and 3899 randomly selected controls aged 50-74 years, whose data were collected through mailed questionnaires. Risk estimates were calculated by unconditional logistic regression. RESULTS: Ever parous women were at reduced risk, with odds ratios of 0.44 (95% confidence interval (CI) 0.26 0.75) for serous and 0.63 (95% CI 0.34-1.19) for mucinous tumors. No clear trends emerged for age at first birth, at menarche, and at menopause. Lactation reduced tumor risk. Oral contraceptive ever use conferred no protection, with odds ratios of 1.40 (95% CI 0.87-2.26) for serous and 1.04 (95% CI 0.61-1.79) for mucinous tumors. The odds ratio for serous tumors following unopposed estrogen ever use was 2.07 (95% CI 1.08-3.95), whereas no risk increase appeared with estrogens supplemented by cyclic or continuous progestins. Mucinous tumors were not associated with hormone replacement therapy. The odds ratio for serous tumors in the highest category of body mass index was 6.47 (95% CI 3.09-13.5). CONCLUSIONS: Increasing parity and lactation reduce the risk of borderline ovarian tumors in women aged 50-74, while no protection follows oral contraceptive use. Hormonal situations such as unopposed estrogen use and obesity, where estrogens are not counteracted by progestins, may increase the risk of serous tumors. PMID- 11733976 TI - BRCA1 germline mutations and polymorphisms in a clinic-based series of ovarian cancer cases: a Gynecologic Oncology Group study. AB - OBJECTIVE: The aims of this study were to determine the frequency of BRCA1 gene alterations in an unselected, clinic-based series of ovarian cancer cases; to evaluate the usefulness of family history in predicting the likelihood of a disease-causing mutation; and to document the occurrence of polymorphic variants in BRCA1 and to determine their distribution among families accordingly to history of breast and/or ovarian cancer. METHOD: Two hundred fifty-eight women with primary epithelial ovarian cancer, entered onto a nonclinical protocol of the Gynecologic Oncology Group, were analyzed for BRCA1 germline alterations by single-strand conformation polymorphism analysis. RESULTS: Protein-truncating mutations in BRCA1 were identified in 12 patients (4.6%). The median age of cancer diagnosis in BRCA1 mutation carriers was 47 years compared to 57 years in patients without mutations (P = 0.02). All but 1 of the patients with BRCA1 mutations reported a family history of breast and/or ovarian cancer and 8 had a first-degree relative with cancer. Twelve mutations of unknown significance were also identified. An association was also noted between the presence of common polymorphisms in BRCA1 and family history of cancer. Polymorphisms were present at higher frequency among women without a family history of cancer compared to women with positive family histories, suggesting they are associated with reduced risk. CONCLUSION: In a clinic-based series of ovarian cancer patients, germline BRCA1 mutations were detected in 12 of 258 (4.6%) patients. A strong correlation was noted between the presence of mutations and family history of breast and/or ovarian cancer, indicating that these women are most likely to benefit from genetic susceptibility testing. PMID- 11733977 TI - Potential clinical utility of CA-125 in responsive but persistent large-volume ovarian cancer following platinum-based chemotherapy. AB - BACKGROUND: Despite the demonstrated clinical utility of the serum CA-125 antigen level in ovarian cancer, controversy exists regarding interpretation of "discordant" results between changes in this tumor marker and measurable disease masses. CASE: A patient with ovarian cancer cared for in the Gynecologic Cancer Program of the Cleveland Clinic Foundation receiving second-line single-agent carboplatin for recurrent disease demonstrated a major response in serum CA-125, but minimal shrinkage of a large painful abdominal mass. A laparotomy was performed both to define the nature of this mass and to attempt to relieve symptoms. The mass was found to be a large "inflamed pseudotumor with central necrosis." No viable tumors cells were found. CONCLUSION: This case represents an excellent example of the remarkably complex biology of malignant disease and suggests how evaluation of changes in CA-125 in women with ovarian cancer may be utilized in individual patients to develop optimal management plans. PMID- 11733978 TI - Diagnosis and localization of testosterone-producing ovarian tumors: imaging or biochemical evaluation. AB - OBJECTIVE: In the testosterone-secreting ovarian tumor (TSOT), the role of whole body positron emission tomography (WBPET) with (fluorine-18)-2-deoxyglucose scanning (FDG) and/or [(11)C]acetate is unclear, although it presents a rationale that these functional tumors would be more active and have increased use of glucose and oxygen consumption than normal tissues. CASE: A 52-year-old woman had a history of steroid cell tumors of the right ovary (IIA) and she received staging surgery including total hysterectomy, salpingo-oophorectomy, and lymph node sampling. Reelevated serum levels of T (5.24 ng/ml) were noted 52 months later. The patient received serial preoperative examinations including WBPET with FDG and acetate, ultrasound, computerized tomography (CT), and magnetic resonance imaging (MRI) to evaluate her recurrence. A suspicious mass on the liver was found on ultrasound, CT, and MRI. The ultrasound-guided biopsy was performed three times, and each of them failed to provide any pathological confirmation. Functional imaging studies showed an abnormal uptake in WBPET using [(11)C]acetate but were negative using FDG. Because of the size of the tumor, the patient's hesitatancy toward an operation, and good previous response to gonadotropin-releasing hormone (GnRH) agonist treatment, the patient received a six-cycle GnRH agonist treatment. Serum T levels returned to normal limits after administration of the first dose of GnRH agonist. At follow-up, serum hormone levels were all within the normal ranges consistent with menopause, but the size of the metastatic tumor was constant. The tumor was then completely excised pathologically proven to be a metastatic TSOT. CONCLUSIONS: Recurrent TSOT might be successfully detected using WBPET with [(11)C]acetate. In addition, GnRH agonist could be tried in patients with TSOT if initial responses were excellent and surgical intervention could not be performed. PMID- 11733979 TI - Paclitaxel and platinum chemotherapy for ovarian carcinoma during pregnancy. AB - BACKGROUND: Ovarian cancer diagnosed during pregnancy is uncommon. Paclitaxel based chemotherapy during pregnancy has not been reported previously. CASE: A woman with ascites and an adnexal mass diagnosed during pregnancy at 27 weeks gestational age underwent a laparotomy with cytoreductive surgery and was diagnosed with stage IIIC papillary serous ovarian adenocarcinoma. She was treated with three cycles of paclitaxel and cisplatin during pregnancy. At 37 weeks, she underwent a cesarean section, abdominal hysterectomy, and cytoreduction. Three additional cycles of chemotherapy were given. She developed a recurrence within 6 weeks of completing chemotherapy. She received several cycles of chemotherapy, but died of recurrent cancer 29 months after diagnosis. The infant has normal growth and development at 30 months of age. CONCLUSION: This is the first reported case of paclitaxel use during pregnancy. PMID- 11733980 TI - Complete remission of gestational choriocarcinoma with choroidal metastasis treated with systemic chemotherapy alone: case report and review of literature. AB - BACKGROUND: Gestational choriocarcinoma is a malignant tumor that frequently metastasizes to the highly vascularized organs such as the lung, brain, and liver via hematogenous spread. However, this tumor rarely metastasizes to the eye and only a few cases of metastasis to the choroid have been reported. CASE: A 17-year old woman presented with visual field defects, decreased vision, and increasing pain in her left eye. She had undergone evacuation of a complete hydatidiform mole 32 months prior to the presentation. Ophthalmologic evaluation revealed a metastatic choroidal tumor, and a CT scan showed a metastatic tumor in the left lung. The serum hCG level was elevated at 7780 mIU/ml. A clinical diagnosis of metastatic gestational choriocarcinoma involving the choroid and lung was made. The patient received 13 courses of combination chemotherapy, resulting in complete remission. Radiotherapy and surgical treatment were unnecessary. CONCLUSION: This is a very rare case of the successful treatment of gestational choriocarcinoma metastatic to the choroid using systemic chemotherapy alone. PMID- 11733981 TI - Ovarian metastasis on transposed ovary in patients treated for squamous cell carcinoma of the uterine cervix: report of two cases and surgical implications. AB - BACKGROUND: To discuss risk factors for ovarian metastasis in a transposed ovary in patients treated for cervical cancer. CASES: Cases were two patients with ovarian metastasis in a transposed ovary in a series of 107 patients. These two patients were treated for a Stage IB squamous cell cervical cancer and presented with a tumor devoid of extrauterine spread (absence of nodal involvement or distant metastases) but with involvement of the uterine corpus. Furthermore, lymphovascular space involvement (LVSI) in the cervix or paracervix was present in both patients. CONCLUSIONS: Ovarian transposition should be performed in patients < or =40 years of age with a small invasive cervical carcinoma (< 3 cm) treated by primary surgery. This procedure should not be performed in patients with bulky tumor and/or in patients with LVSI. PMID- 11733982 TI - Patent foramen ovale complicated by paradoxical embolism and brain infarct in a patient with advanced ovarian cancer. AB - BACKGROUND: Recent investigations of patients with cerebral and peripheral arterial emboli of unknown cause suggest that paradoxical embolism through a patent foramen ovale might be responsible for more arterial embolic events than previously realized. CASE: A 60-year-old woman with advanced ovarian cancer presented with sudden onset of expressive aphasia and right upper hemiplegia postoperatively. A patent foramen ovale diagnosed by echocardiography with contrast combined with the presence of thrombosis in her right femoral vein leads us to speculate that her stroke was secondary to a paradoxical embolism. CONCLUSION: Paradoxical embolism should be considered in the differential diagnosis of ovarian cancer patients with embolic stroke and it may be appropriate to include a cardiac echo as part of the diagnostic evaluation. PMID- 11733983 TI - Liposarcoma complicating pregnancy. AB - BACKGROUND: An adnexal mass in pregnancy is a common occurrence. Malignancy is uncommon and only seven cases of liposarcoma have been reported in association with pregnancy. CASE: We present a case of a pregnant patient who was diagnosed with an adnexal mass detected by ultrasound at 13 weeks gestation. The mass was surgically removed postpartum, when it was found to be a liposarcoma. Postoperatively she received chemotherapy but eventually succumbed to her disease. CONCLUSIONS: Liposarcoma is rarely found in association with pregnancy and may follow an aggressive course. PMID- 11733984 TI - Inflammatory breast metastases of ovarian cancer: a case report. AB - BACKGROUND: Metastasis to the breast from extramammary malignancies is rare. CASE: A 35-year-old woman presented with bilaterally inflammatory breast involvement, 2 years after the diagnosis of stage IIIC epithelial ovarian cancer. Neoplastic tissue was immunohistochemically positive using antibodies against OC125 and negative for gross cystic disease fluid protein-15 (BRST-2) and estrogen receptor in biopsy material in the breast. Combination chemotherapy consisting of paclitaxel, cisplatin, and anthracycline was started. She died 18 months after the breast metastasis. CONCLUSION: Ovarian carcinoma usually presents with signs and symptoms related to the tumor burden within the abdominal cavity. The finding of isolated, distant metastases such as breast involvement without intraabdominal disease is extremely rare. Determining the origin of the primary tumor is important in directing the actual therapy. PMID- 11733985 TI - Modifying the specificity of an RNA backbone contact. AB - The interaction between the MS2 bacteriophage coat protein homodimer and its cognate RNA hairpin is facilitated by 21 different RNA-protein contacts. In one of these contacts, the 2'-hydroxyl group at ribose -5 of the RNA acts as a hydrogen bond donor to Glu63 in one subunit of the protein. Previous experiments showed that substitution of ribose -5 with deoxyribose resulted in a 24-fold decrease in binding affinity between RNA and protein. Using a protein where the two MS2 monomers were fused to increase stability, the contribution of this contact to the overall binding affinity was investigated by site-directed mutagenesis. When Glu63 was substituted with glutamine, aspartate, or alanine, the binding affinity of the hairpin for the protein was weakened by 12 to 100 fold, similar to that observed with deoxyribose at position -5. However, the specificity of the three mutant proteins for RNAs with various modifications at the 2'-position of ribose -5 differed dramatically. While the Glu63Asp protein resembled the wild-type protein in preferring the 2'-hydroxyl group over a proton or a bulky 2'-substituent, both the Glu63Ala and Glu63Gln proteins preferred bulky 2'-substituents over the 2'-hydroxyl group by more than 100-fold. These experiments emphasize the ease with which the specificity of a protein-nucleic acid interaction can be changed at thermodynamically important sites. PMID- 11733986 TI - Three-dimensional structures of the Mn and Mg dTDP complexes of the family GT-2 glycosyltransferase SpsA: a comparison with related NDP-sugar glycosyltransferases. AB - The vast majority of glycosidic-bond synthesis in nature is performed by glycosyltransferases, which use activated glycosides as the sugar donor. Typically, the activated leaving group is a nucleoside phosphate, lipid phosphate or phosphate. The nucleotide-sugar-dependent glycosyltransferases fall into over 50 sequence-based families, with the largest and most widespread family of inverting transferases named family GT-2. Here, we present the three-dimensional crystal structure of SpsA, the first and currently the only structural representative from family GT-2, in complex with both Mn-dTDP and Mg-dTDP at a resolution of 2 A. These structures reveal how SpsA and related enzymes may display nucleotide plasticity and permit a comparison of the catalytic centre of this enzyme with those from related sequence families whose three-dimensional structures have recently been determined. Family GT-2 enzymes, together with enzymes from families 7, 13 and 43, appear to form a clan of related structures with identical catalytic apparatus and reaction mechanism. PMID- 11733987 TI - Enhanced crystallization of the Cys18 to Ser mutant of bovine gammaB crystallin. AB - The cysteine residues of the gamma crystallins, a family of ocular lens proteins, are involved in the aggregation and phase separation of these proteins. Both these phenomena are implicated in cataract formation. We have used bovine gammaB crystallin as a model system to study the role of the individual cysteine residues in the aggregation and phase separation of the gamma crystallins. Here, we compare the thermodynamic and kinetic behavior of the recombinant wild-type protein (WT) and the Cys18 to Ser (C18S) mutant. We find that the solubilities of the two proteins are similar. The kinetics of crystallization, however, are different. The WT crystallizes slowly enough for the metastable liquid-liquid coexistence to be easily observed. C18S, on the other hand, crystallizes rapidly; the metastable coexisting liquid phases of the pure mutant do not form. Nevertheless, the coexistence curve of C18S can be determined provided that crystallization is kinetically suppressed. In this way we found that the coexistence curve coincides with that of the WT. Despite the difference in the kinetics of crystallization, the two proteins were found to have the same crystal forms and almost identical X-ray structures. Our results demonstrate that even conservative point mutations can bring about dramatic changes in the kinetics of crystallization. The implications of our findings for cataract formation and protein crystallization are discussed. PMID- 11733988 TI - Dimer stabilization upon activation of the transcriptional antiterminator LicT. AB - LicT belongs to the BglG/SacY family of transcriptional antiterminators that induce the expression of sugar metabolizing operons in Gram positive and Gram negative bacteria. These proteins contain a N-terminal RNA-binding domain and a regulatory domain called PRD which is phosphorylated on conserved histidine residues by components of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Although it is now well established that phosphorylation of PRD-containing transcriptional regulators tunes their functional response, the molecular and structural basis of the regulation mechanism remain largely unknown.A constitutively active LicT variant has been obtained by introducing aspartic acid in replacement of His207 and His269, the two phosphorylatable residues of the PRD2 regulatory sub-domain. Here, the functional and structural consequences of these activating mutations have been evaluated in vitro using various techniques including surface plasmon resonance, limited proteolysis, analytical centrifugation and X-ray scattering. Comparison with the native, unphosphorylated form shows that the activating mutations enhance the RNA-binding activity and induce tertiary and quaternary structural changes. Both mutant and native LicT form dimers in solution but the native dimer exhibits a less stable and more open conformation than the activated mutant form. Examination of the recently determined crystal structure of mutant LicT regulatory domain suggests that dimer stabilization is accomplished through salt-bridge formation at the PRD2:PRD2 interface, resulting in domain motion and dimer closure propagating the stabilizing effect from the protein C-terminal end to the N-terminal effector domain. These results suggest that LicT activation arises from a conformational switch inducing long range rearrangement of the dimer interaction surface, rather than from an oligomerization switch converting an inactive monomer into an active dimer. PMID- 11733989 TI - Purification and characterization of the 1.0 MDa CCR4-NOT complex identifies two novel components of the complex. AB - The CCR4-NOT complex is an evolutionarily conserved, transcriptional regulatory complex that is involved in controlling mRNA initiation, elongation and degradation. The CCR4-NOT proteins from Saccharomyces cerevisiae exist in two complexes, 1.9x10(6) Da and 1.0x10(6) Da (1.0 MDa) in size, and individual components of these complexes display such disparate functions as binding to and restricting TFIID functions, contacting SAGA and contributing to mRNA deadenylation. As a first step in characterizing the functional roles of the 1.0 MDa complex, we have purified it to near homogeneity. Mass spectrometric analysis was subsequently used to identify all the components of the complex. The 1.0 MDa complex was found to contain CCR4, CAF1, NOT1-5 and two new proteins, CAF40 and CAF130. CAF130 and CAF40 are two unique yeast proteins, with CAF40 displaying extensive homology to proteins from other eukaryotes. Immunoprecipitation and gel filtration experiments confirmed that CAF130 and CAF40 are components of both of the 1.9 MDa and 1.0 MDa CCR4-NOT complexes. Biochemical analysis indicated that the CAF40 and CAF130 proteins bind to the NOT1 protein and exist in a location separate from the two other subsets of proteins in the complex: the CCR4 and CAF1 proteins, and the NOT2, NOT4 and NOT5 proteins. Moreover, CAF40 was able to interact with human NOT1, suggesting that human CAF40 would also be a component of the recently identified human CCR4-NOT complex. Analysis of caf40 and caf130 deletions indicated that they elicited phenotypes shared by defects in other CCR4 NOT genes. The distinct location of CAF40 and CAF130 and the evolutionary conservation of CAF40 implicate them in novel roles in the function of the CCR4 NOT complex. PMID- 11733990 TI - Distinctive features of the two classes of eukaryotic peptide deformylases. AB - Peptide deformylases (PDFs) are essential enzymes of the N-terminal protein processing pathway of eubacteria. The recent discovery of two types of PDFs in higher plants, PDF1A and PDF1B, and the detection of PDF1A in humans, have raised questions concerning the importance of deformylation in eukaryotes. Here, we have characterized fully in vitro and compared the properties of the two classes of eukaryotic PDFs, PDF1A and PDF1B, using the PDFs from Arabidopsis thaliana and Lycopersicon esculentum. We have shown that the PDFs of a given class (1A or 1B) all display similar features, independently of their origin. We also observed similar specificity of all plant PDFs for natural substrate peptides, but identified a number of biochemical differences between the two classes (1A or 1B). The main difference lies at the level of the bound cofactor, iron for PDF1B like bacterial PDFs, and zinc for PDF1A. The nature of the metal cation has important consequences concerning the relative sensitivity to oxygen of the two plant PDFs. Investigation of the specificity of these enzymes with unusual substrates revealed additional differences between the two types of PDFs, enabling us to identify specific inhibitors with a lower affinity against PDF1As. However, the two plant PDFs were inhibited equally strongly in vitro by actinonin, an antibiotic that specifically acts on bacterial PDFs. Uptake of actinonin by A. thaliana seedlings was used to investigate the function of PDFs in the plant. Because it induces an albino phenotype, we conclude that deformylation is likely to play an essential role in the chloroplast. PMID- 11733991 TI - Expansion/contraction of mammalian mitochondrial DNA repeats in Escherichia coli mimics the mitochondrial heteroplasmy. AB - Length polymorphism due to tandem repeats is a common feature in animal mitochondrial DNA. The rabbit mitochondrial genome contains a 20 bp repeat domain, which generates a general heteroplasmic state. The observed polymorphic patterns suggest a dynamic equilibrium between gain and loss of units that maintains the copy number in the range 3-19 repeat units. In the apparent absence of recombination, slipped-strand mispairing during replication appears to be the primary cause of additions and deletions. To investigate this hypothesis we have set up a plasmid assay in Escherichia coli. A variable number of repeat units was inserted into a plasmid in both orientations relative to the colE1 origin of replication. Our data show that (i) a minimum unit number (>3) is necessary to generate length polymorphs, (ii) the number of events increases with the length tract, (iii) an excess of additions over deletions is found when the copy number is less than 10 and the trend is reversed when it is over 10, (iv) the frequency of deletions-additions is dependent on the orientation, (v) the polymorphism patterns are different according to the orientation. The length polymorphic pattern generated in the bacteria, in one orientation, mimics that observed in the mitochondria, suggesting that slipped mispairing between repeated sequences during DNA replication is responsible for the mitochondrial heteroplasmic state. PMID- 11733992 TI - DNA recombination and RNA cleavage activities of the Flp protein: roles of two histidine residues in the orientation and activation of the nucleophile for strand cleavage. AB - Using a combination of DNA and hybrid DNA-RNA substrates, we have analyzed the mechanism of phosphoryl transfer by the Flp site-specific recombinase in three different reactions: DNA strand breakage and joining, and two types of RNA cleavage activities. These reactions were then used to characterize Flp variants altered at His309 and His345, amino acid residues that are in close proximity to two key catalytic residues (Arg308 and Tyr343). These histidine residues are important for strand cutting by Tyr343, the active-site nucleophile of Flp, but neither residue contributes to the type II RNA cleavage activity or to the strand joining reaction in a pre-cleaved substrate. Strand cleavage reactions using small, diffusible nucleophiles indicate that this histidine pair contributes to the correct positioning and activation of Tyr343 within the shared active site of Flp. The implications of these results are evaluated against the recently solved crystal structure of Flp in association with a Holliday junction. PMID- 11733993 TI - Mechanism and cleavage specificity of the H-N-H endonuclease colicin E9. AB - Colicin endonucleases and the H-N-H family of homing enzymes share a common active site structural motif that has similarities to the active sites of a variety of other nucleases such as the non-specific endonuclease from Serratia and the sequence-specific His-Cys box homing enzyme I-PpoI. In contrast to these latter enzymes, however, it remains unclear how H-N-H enzymes cleave nucleic acid substrates. Here, we show that the H-N-H enzyme from colicin E9 (the E9 DNase) shares many of the same basic enzymological characteristics as sequence-specific H-N-H enzymes including a dependence for high concentrations of Mg2+ or Ca2+ with double-stranded substrates, a high pH optimum (pH 8-9) and inhibition by monovalent cations. We also show that this seemingly non-specific enzyme preferentially nicks double-stranded DNA at thymine bases producing 3'-hydroxy and 5'-phosphate termini, and that the enzyme does not cleave small substrates, such as dinucleotides or nucleotide analogues, which has implications for its mode of inhibition in bacteria by immunity proteins. The E9 DNase will also bind single-stranded DNA above a certain length and in a sequence-independent manner, with transition metals such as Ni2+ optimal for cleavage but Mg2+ a poor cofactor. Ironically, the H-N-H motif of the E9 DNase although resembling the zinc binding site of a metalloenzyme does not support zinc-mediated hydrolysis of any DNA substrate. Finally, we demonstrate that the E9 DNase also degrades RNA in the absence of metal ions. In the context of current structural information, our data show that the H-N-H motif is an adaptable catalytic centre able to hydrolyse nucleic acid by different mechanisms depending on the substrate and metal ion regime. PMID- 11733994 TI - Short arm region of laminin-5 gamma2 chain: structure, mechanism of processing and binding to heparin and proteins. AB - Laminin-5 is a typical component of several epithelial tissues and contains a unique gamma2 chain which can be proteolytically processed by BMP-1. This occurs in the N-terminal half of the gamma2 chain (606 residues), which consists of two rod-like tandem arrays of LE modules, LE1-3 and LE4-6, that flank a globular L4m module containing the cleavage site. Recombinant analysis of L4m, which includes an additional imperfect LE module essential for proper folding, demonstrated an unusual pattern of disulfide bonding. These connectivities prevented the release of gamma2LE1-3L4 m after BMP-1 cleavage which required in addition disulfide reshuffling by isomerases. The liberated segment bound through its L4 m module to heparin, nidogen-1, fibulin-1 and fibulin-2. A further heparin/sulfatide-binding site could be attributed to some arginine residues in module LE1. The gamma2LE4-6 segment remaining in processed laminin-5 showed only a strong binding to fibulin 2. Immunological studies showed a similar partial processing in cell culture and tissues and the persistence of the released fragment in tissues. This indicated that both N-terminal regions of the gamma2 chain may have a function in vivo. PMID- 11733995 TI - Tumor cell growth inhibition and extracellular signal-regulated kinase (ERK) phosphorylation by novel K vitamins. AB - 2-(2-hydroxy-ethylsulfanyl)-3-methyl-1,4-naphthoquinone or CPD-5, a K vitamin analog, was previously indicated to be a potent growth inhibitor for Hep 3B hepatoma cells in vitro. Here, we show that CPD-5 and two newly synthesized analogs, 2-(2-hydroxy-ethylsulfanyl)-3-methyl-5- nitro-1,4-naphthoquinone (PD-37) and 2-(2-hydroxy-ethylsulfanyl)-3- methyl-5-acetylamino-1,4-naphthoquinone (PD 42), are potent growth inhibitors of 13 different human cancer cell lines, with IC50 values in the range of 3-54 microM. Phospho-ERK was induced by each of three K vitamin analogs in every cell line in a dose-dependent manner, at growth inhibitory doses. ERK phosphorylation and growth inhibitory effects were strongly correlated, with p=0.0080 for CPD-5, p=0.0076 for PD-37 and p=0.0251 for PD-42. The induction of phospho-ERK and growth inhibition were antagonized by thiol containing anti-oxidants, but not by catalase, consistent with a possible arylating mechanism. The data show a novel class of growth inhibitors with a wide spectrum of action that induces ERK hyper-phosphorylation, as a possible new growth inhibitory feature. PMID- 11733996 TI - Structure and properties of a dimeric N-terminal fragment of human ubiquitin. AB - Previous peptide dissection and kinetic experiments have indicated that in vitro folding of ubiquitin may proceed via transient species in which native-like structure has been acquired in the first 45 residues. A peptide fragment, UQ(1 51), encompassing residues 1 to 51 of ubiquitin was produced in order to test whether this portion has propensity for independent self-assembly. Surprisingly, the construct formed a folded symmetrical dimer that was stabilised by 0.8 M sodium sulphate at 298 K (the S state). The solution structure of the UQ(1-51) dimer was determined by multinuclear NMR spectroscopy. Each subunit of UQ(1-51) consists of an N-terminal beta-hairpin followed by an alpha-helix and a final beta-strand, with orientations similar to intact ubiquitin. The dimer is formed by the third beta-strand of one subunit interleaving between the hairpin and third strand of the other to give a six-stranded beta-sheet, with the two alpha helices sitting on top. The helix-helix and strand portions of the dimer interface also mimic related features in the structure of ubiquitin. The structural specificity of the UQ(1-51) peptide is tuneable: as the concentration of sodium sulphate is decreased, near-native alternative conformations are populated in slow chemical exchange. Magnetization transfer experiments were performed to characterize the various species present in 0.35 M sodium sulphate, namely the S state and two minor forms. Chemical shift differences suggest that one minor form is very similar to the S state, while the other experiences a significant conformational change in the third strand. A segmental rearrangement of the third strand in one subunit of the S state would render the dimer asymmetric, accounting for most of our results. Similar small-scale transitions in proteins are often invoked to explain solvent exchange at backbone amide proton sites that have an intermediate level of protection. PMID- 11733997 TI - Crystal structure of omega transcriptional repressor encoded by Streptococcus pyogenes plasmid pSM19035 at 1.5 A resolution. AB - The 71 amino acid residue omega protein encoded by the Streptococcus pyogenes non conjugative plasmid pSM19035 is a transcriptional repressor that regulates expression of genes for copy number control and stable maintenance of plasmids. The crystal structure of omega protein has been determined by multiple isomorphous replacement, including anomalous scattering and refined to an R factor of 21.1 % (R(free)=23.2 %) at 1.5 A resolution. Two monomers related by a non-crystallographic 2-fold axis form a homodimer that occupies the asymmetric unit. Each polypeptide chain is folded into two alpha-helices and one beta-strand forming an antiparallel beta-ribbon in the homodimer. The N-terminal regions (1 23 and 1-22 in subunits I and II, respectively) are not defined in the electron density due to proteolysis of the N-terminal 20 amino acid residues during crystallisation and partial disorder. The omega protein belongs to the structural superfamily of MetJ/Arc repressors featuring a ribbon-helix-helix DNA-binding motif with the beta-ribbon located in and recognizing the major groove of operator DNA; according to a modelled omega protein-DNA complex, residues Arg31 and Arg31' on the beta-ribbon are in positions to interact with a nucleobase, especially guanine. PMID- 11733998 TI - Recognition of cello-oligosaccharides by a family 17 carbohydrate-binding module: an X-ray crystallographic, thermodynamic and mutagenic study. AB - The crystal structure of the Clostridium cellulovorans carbohydrate-binding module (CBM) belonging to family 17 has been solved to 1.7 A resolution by multiple anomalous dispersion methods. CBM17 binds to non-crystalline cellulose and soluble beta-1,4-glucans, with a minimal binding requirement of cellotriose and optimal affinity for cellohexaose. The crystal structure of CBM17 complexed with cellotetraose solved at 2.0 A resolution revealed that binding occurs in a cleft on the surface of the molecule involving two tryptophan residues and several charged amino acids. Thermodynamic binding studies and alanine scanning mutagenesis in combination with the cellotetraose complex structure allowed the mapping of the CBM17 binding cleft. In contrast to the binding groove characteristic of family 4 CBMs, family 17 CBMs appear to have a very shallow binding cleft that may be more accessible to cellulose chains in non-crystalline cellulose than the deeper binding clefts of family 4 CBMs. The structural differences in these two modules may reflect non-overlapping binding niches on cellulose surfaces. PMID- 11733999 TI - Crystal structure of an antigen-binding fragment bound to single-stranded DNA. AB - Antibodies to DNA are characteristic of the autoimmune disease systemic lupus erythematosus (SLE) and they also serve as models for the study of protein-DNA recognition. Anti-DNA antibodies often play an important role in disease pathogenesis by mediating kidney damage via antibody-DNA immune complex formation. The structural underpinnings of anti-DNA antibody pathogenicity and antibody-DNA recognition, however, are not well understood, due in part to the lack of direct, experimental three-dimensional structural information on antibody DNA complexes. To address these issues for anti-single-stranded DNA antibodies, we have determined the 2.1 A crystal structure of a recombinant Fab (DNA-1) in complex with dT5. DNA-1 was previously isolated from a bacteriophage Fab display library from the immunoglobulin repertoire of an SLE-prone mouse. The structure shows that DNA-1 binds oligo(dT) primarily by sandwiching thymine bases between Tyr side-chains, which allows the bases to make sequence-specific hydrogen bonds. The critical stacking Tyr residues are L32, L49, H100, and H100A, while His L91 and Asn L50 contribute hydrogen bonds. Comparison of the DNA-1 structure to other anti-nucleic acid Fab structures reveals a common ssDNA recognition module consisting of Tyr L32, a hydrogen bonding residue at position L91, and an aromatic side-chain from the tip of complementarity determining region H3. The structure also provides a framework for interpreting previously determined thermodynamics data, and this analysis suggests that hydrophobic desolvation might underlie the observed negative enthalpy of binding. Finally, Arg side chains from complementarity determining region H3 appear to play a novel role in DNA-1. Rather than forming ion pairs with dT5, Arg contributes to oligo(dT) recognition by helping to maintain the structural integrity of the combining site. This result is significant because antibody pathogenicity is thought to be correlated to the Arg content of anti-DNA antibody hypervariable loops. PMID- 11734001 TI - Solution NMR structure of the myosin phosphatase inhibitor protein CPI-17 shows phosphorylation-induced conformational changes responsible for activation. AB - Contractility of vascular smooth muscle depends on phosphorylation of myosin light chains, and is modulated by hormonal control of myosin phosphatase activity. Signaling pathways activate kinases such as PKC or Rho-dependent kinases that phosphorylate the myosin phosphatase inhibitor protein called CPI 17. Phosphorylation of CPI-17 at Thr38 enhances its inhibitory potency 1000-fold, creating a molecular on/off switch for regulating contraction. We report the solution NMR structure of the CPI-17 inhibitory domain (residues 35-120), which retains the signature biological properties of the full-length protein. The final ensemble of 20 sets of NMR coordinates overlaid onto their mean structure with r.m.s.d. values of 0.84(+/-0.22) A for the backbone atoms. The protein forms a novel four-helix, V-shaped bundle comprised of a central anti-parallel helix pair (B/C helices) flanked by two large spiral loops formed by the N and C termini that are held together by another anti-parallel helix pair (A/D helices) stabilized by intercalated aromatic and aliphatic side-chains. Chemical shift perturbations indicated that phosphorylation of Thr38 induces a conformational change involving displacement of helix A, without significant movement of the other three helices. This conformational change seems to flex one arm of the molecule, thereby exposing new surfaces of the helix A and the nearby phosphorylation loop to form specific interactions with the catalytic site of the phosphatase. This phosphorylation-dependent conformational change offers new structural insights toward understanding the specificity of CPI-17 for myosin phosphatase and its function as a molecular switch. PMID- 11734000 TI - Refined crystallographic structure of Pseudomonas aeruginosa exotoxin A and its implications for the molecular mechanism of toxicity. AB - Exotoxin A of Pseudomonas aeruginosa asserts its cellular toxicity through ADP ribosylation of translation elongation factor 2, predicated on binding to specific cell surface receptors and intracellular trafficking via a complex pathway that ultimately results in translocation of an enzymatic activity into the cytoplasm. In early work, the crystallographic structure of exotoxin A was determined to 3.0 A resolution, revealing a tertiary fold having three distinct structural domains; subsequent work has shown that the domains are individually responsible for the receptor binding (domain I), transmembrane targeting (domain II), and ADP-ribosyl transferase (domain III) activities, respectively. Here, we report the structures of wild-type and W281A mutant toxin proteins at pH 8.0, refined with data to 1.62 A and 1.45 A resolution, respectively. The refined models clarify several ionic interactions within structural domains I and II that may modulate an obligatory conformational change that is induced by low pH. Proteolytic cleavage by furin is also obligatory for toxicity; the W281A mutant protein is substantially more susceptible to cleavage than the wild-type toxin. The tertiary structures of the furin cleavage sites of the wild-type and W281 mutant toxins are similar; however, the mutant toxin has significantly higher B factors around the cleavage site, suggesting that the greater susceptibility to furin cleavage is due to increased local disorder/flexibility at the site, rather than to differences in static tertiary structure. Comparison of the refined structures of full-length toxin, which lacks ADP-ribosyl transferase activity, to that of the enzymatic domain alone reveals a salt bridge between Arg467 of the catalytic domain and Glu348 of domain II that restrains the substrate binding cleft in a conformation that precludes NAD+ binding. The refined structures of exotoxin A provide precise models for the design and interpretation of further studies of the mechanism of intoxication. PMID- 11734002 TI - Flexibility and nucleation in sickle hemoglobin. AB - We have studied the self-assembly of Hemoglobin C-Harlem (HbC-Harlem), a double mutant of hemoglobin that possesses the beta6 Glu-->Val mutation of sickle hemoglobin (HbS) plus beta73 Asp-->Asn. By electron microscopy we find it forms crystals, rather than the wrapped multistranded fibers seen in HbS. Fourier transforms of the crystals yield unit cell parameters indistinguishable from crystals of HbS. Differential interference contrast (DIC) microscopy and birefringence also show crystal formation rather than the polymers or domains seen for HbS, while the growth patterns showed radiating crystal structures rather than simple linear crystalline forms. The solubility of the assembly was measured using a photolytic micromethod over a temperature range of 17-31 degrees C in 0.15 M phosphate buffer and found to be essentially the same as that of fibers of HbS. The assembly kinetics were observed by photolysis of the carbon monoxide derivative, and the mass of assembled hemoglobin was found to grow exponentially, with onset times that were stochastically distributed for small volumes. The stochastic onset of assembly showed strong concentration dependence, similar to but slightly greater than that seen in sickle hemoglobin nucleation. These observations suggest that like HbS, HbC-Harlem assembly proceeds by a homogeneous nucleation process, followed by heterogeneous nucleation. However, relative to HbS, both homogeneous and heterogeneous nucleation are suppressed by almost 11 orders of magnitude. The slowness of nucleation can be reconciled with the similarity of the solubility to HbS by an increase in contact energy coupled with a decrease in vibrational entropy recovered on assembly. This also explains the linearity of the double-strands, and agrees with the chemical nature of the structural replacement. PMID- 11734003 TI - Native-state energetics of a thermostabilized variant of ribonuclease HI. AB - Escherichia coli RNase HI is a well-characterized model system for protein folding and stability. Controlling protein stability is critical for both natural proteins and for the development of engineered proteins that function under extreme conditions. We have used native-state hydrogen exchange on a variant containing the stabilizing mutation Asp10 to alanine in order to determine its residue-specific stabilities. On average, the DeltaG(unf) value for each residue was increased by 2-3 kcal/mol, resulting in a lower relative population of partially unfolded forms. Though increased in stability by a uniform factor, D10A shows a distribution of stabilities in its secondary structural units that is similar to that of E. coli RNase H, but not the closely related protein from Thermus thermophilus. Hence, the simple mutation used to stabilize the enzyme does not recreate the balance of conformational flexibility evolved in the thermophilic protein. PMID- 11734004 TI - Characterization of pH-induced transitions of beta-lactoglobulin: ultrasonic, densimetric, and spectroscopic studies. AB - Depending on solution conditions, beta-lactoglobulin can exist in one of its six pH-dependent structural states. We have characterized the acid and basic-induced conformational transitions between these structural states over the pH range of pH 1 to pH 13. To this end, we have employed high-precision ultrasonic and densimetric measurements coupled with fluorescence and CD spectroscopic data. Our combined spectroscopic and volumetric results have revealed five pH-induced transitions of beta-lactoglobulin between pH 1 and pH 13. The first transition starts at pH 2 and is not completed even at pH 1, our lowest experimental pH. This transition is followed by the dimer-to-monomer transition of beta lactoglobulin between pH 2.5 and pH 4. The dimer-to-monomer transition is accompanied by decreases in volume, v degrees (-0.008(+/-0.003) cm3 x g(-1)), and adiabatic compressibility, k degrees (S) (-(0.7(+/-0.4))x10(-6) cm3 x g(-1) x bar(-1)). We interpret the observed changes in volume and compressibility associated with the dimer-to-monomer transition of beta-lactoglobulin, in conjunction with X-ray crystallographic data, as suggesting a 7 % increase in protein hydration, with the hydration changes being localized in the area of contact between the two monomeric subunits. The so-called N-to-Q transition of beta-lactoglobulin occurs between pH 4.5 and pH 6 and is accompanied by increases in volume, v degrees (0.004(+/-0.003) cm3 x g(-1)), and compressibility, k degrees (S) ((0.7(+/-0.4))x10(-6) cm3 x g(-1) x bar(-1)). The Tanford transition of beta-lactoglobulin is centered at pH 7.5 and is accompanied by a decrease in volume, v degrees (-0.006(+/-0.003) cm3 x g(-1)), and an increase in compressibility, k degrees (S) ((1.5(+/-0.5))x10(-6) cm3 x g(-1) x bar(-1)). Based on these volumetric results, we propose that the Tanford transition is accompanied by a 5 to 10 % increase in the protein hydration and a loosening of the interior packing of beta-lactoglobulin as reflected in a 12 % increase in its intrinsic compressibility. Finally, above pH 9, the protein undergoes irreversible base-induced unfolding which is accompanied by decreases in v degrees (-0.014(+/-0.003) cm3 x g(-1)) and k degrees (S) (-(7.0(+/-0.5))x10(-6) cm3 x g(-1) x bar(-1)). Combining these results with our CD spectroscopic data, we propose that, in the base-induced unfolded state of beta-lactoglobulin, only 80 % of the surface area of the fully unfolded conformation is exposed to the solvent. Thus, in so far as solvent exposure is concerned, the base-induced unfolded states of beta-lactoglobulin retains some order, with 20 % of its amino acid residues remaining solvent inaccessible. PMID- 11734005 TI - Folding of circular permutants with decreased contact order: general trend balanced by protein stability. AB - To examine the influence of contact order and stability on the refolding rate constant for two-state proteins, we have analysed the folding kinetics of the small beta-alpha-beta protein S6 and two of its circular permutants with relative contact orders of 0.19, 0.15 and 0.12. Data reveal a small but significant increase of the refolding rate constant (log k(f)) with decreasing contact order. At the same time, the decreased contact order is correlated to losses in global stability and alterations of the folding nucleus. When the differences in stability are accounted for by addition of Na2SO4 or by comparison of the folding kinetics at the transition mid-point, the dependence between log k(f) and contact order becomes stronger and follows the general correlation for two-state proteins. The observation emphasizes the combined action of topology and stability in controlling the rate constant of protein folding. PMID- 11734006 TI - Importance of two ATP-binding sites for oligomerization, ATPase activity and chaperone function of mitochondrial Hsp78 protein. AB - The yeast mitochondrial chaperone Hsp78, a homologue of yeast cytosolic Hsp104 and bacterial ClpB, is required for maintenance of mitochondrial functions under heat stress. Here, Hsp78 was purified to homogeneity and shown to form a homo hexameric complex, with an apparent molecular mass of approximately 440 kDa, in an ATP-dependent manner. Analysis of its ATPase activity reveals that the observed positive cooperativity effect depends both on Hsp78 and ATP concentration. Site-directed mutagenesis of the two putative Hsp78 nucleotide binding domains suggest that the first nucleotide-binding domain is responsible for ATP hydrolysis and the second one for protein oligomerization. Studies on the chaperone activity of Hsp78 show that its cooperation with the mitochondrial Hsp70 system, consisting of Ssc1p, Mdj1p and Mge1p, is needed for the efficient reactivation of substrate proteins. These studies also suggest that the oligomerization but not the Hsp78 ATPase activity is essential for its chaperone activity. PMID- 11734007 TI - Crowding and hydration effects on protein conformation: a study with sol-gel encapsulated proteins. AB - We are developing an experimental system for testing the effects of macromolecular crowding and molecular confinement on protein structure. In the present study, solvent effects on the secondary structure of two proteins were examined by circular dichroism following encapsulation in the hydrated pores of a silica glass matrix by the sol-gel method. Changes in the unfolded conformations of encapsulated apomyoglobin and reduced serum albumin were analyzed after equilibration with aqueous solutions of natural osmolytes, short-chain alcohols, polyethylene glycol, and a complete series of Hofmeister cations. In many instances, the alpha-helical content of the encapsulated protein was increased by addition of solutes at concentrations that have no effect on the protein in the absence of the glass. The results are discussed from the perspective of water structure. We argue that perturbed water at the silica interface causes an increase in the average free energy of the bulk water phase which, consequently, diminishes the strength of the hydrophobic effect inside the glass matrix and destabilizes the conformation of encapsulated proteins. We propose that solutes can increase the strength of the hydrophobic effect and influence folding equilibria without directly interacting with the protein. A hypothesis is provided for the apparent paradox that kosmotropic (strongly water binding) anions favor native protein structure, whereas chaotropic (weakly water binding) cations enhance native protein structure. The encapsulation results suggest that macromolecular crowding and molecular confinement are accompanied by hydration effects that may oppose or potentiate the stabilizing effects of excluded volume on protein structure, depending on the surface chemistry of the crowding agent and its influence on bulk water structure. In the crowded environment of a living cell, excluded volume effects, surface-induced water structure, and compatible solutes are expected to complement the dominant forces in protein folding. PMID- 11734008 TI - [L29M] substitution in the interface of subunit-subunit interactions enhances Escherichia coli RecA protein properties important for its recombinogenic activity. AB - Genetic analysis of RecA protein chimeras and their ancestors, RecAEc (from Escherichia coli) and RecAPa (Pseudomonas aeruginosa) had allowed us to place these proteins with respect to their recombinogenic activities in the following order: RecAPa>RecAX21>RecAX20=RecAEc. While RecAX20 differs from RecAEc in five amino acid residues with two substitutions ([S25A] and [I26V]) at the interface of subunit interactions in the RecA polymer, RecAX20 and RecAX21 differ only by a single substitution [L29M] present at the interface. Here, we present an analysis of the biochemical properties considered important for the recombinogenic activity of all four RecA proteins. While RecAX20 was very similar to RecAEc by all activities analysed, RecAX21 differed from RecAEc in several respects. These differences included an increased affinity for double-stranded DNA, a more active displacement of SSB protein from single-stranded DNA (ssDNA), a decreased end dependent RecAX21 protein dissociation from a presynaptic complex, and a greater accumulation of intermediate products relative to the final product in the strand exchange reaction. RecAPa was more tolerant than RecAX21 only to the end dependent RecA protein dissociation. In addition, RecAPa was more resistant to temperature and salt concentrations in its ability to form a presynaptic RecAPa::ATP::ssDNA filament. Calculations of conformational energy revealed that the [L29M] substitution in RecAX21 polymer caused an increase in its flexibility. This led us to conclude that even a small change in the flexibility of the RecA presynaptic complex could profoundly affect its recombinogenic properties. PMID- 11734009 TI - Towards a reliable objective function for multiple sequence alignments. AB - Multiple sequence alignment is a fundamental tool in a number of different domains in modern molecular biology, including functional and evolutionary studies of a protein family. Multiple alignments also play an essential role in the new integrated systems for genome annotation and analysis. Thus, the development of new multiple alignment scores and statistics is essential, in the spirit of the work dedicated to the evaluation of pairwise sequence alignments for database searching techniques. We present here norMD, a new objective scoring function for multiple sequence alignments. NorMD combines the advantages of the column-scoring techniques with the sensitivity of methods incorporating residue similarity scores. In addition, norMD incorporates ab initio sequence information, such as the number, length and similarity of the sequences to be aligned. The sensitivity and reliability of the norMD objective function is demonstrated using structural alignments in the SCOP and BAliBASE databases. The norMD scores are then applied to the multiple alignments of the complete sequences (MACS) detected by BlastP with E-value<10, for a set of 734 hypothetical proteins encoded by the Vibrio cholerae genome. Unrelated or badly aligned sequences were automatically removed from the MACS, leaving a high quality multiple alignment which could be reliably exploited in a subsequent functional and/or structural annotation process. After removal of unreliable sequences, 176 (24 %) of the alignments contained at least one sequence with a functional annotation. 103 of these new matches were supported by significant hits to the Interpro domain and motif database. PMID- 11734010 TI - Alamethicin-leucine zipper hybrid peptide: a prototype for the design of artificial receptors and ion channels. AB - In this report, we describe a novel concept of extramembrane control of channel peptide assembly and the eventual channel current modulation. Alamethicin is a peptide antibiotic, which usually forms ion channels in various association states. By introducing an extramembrane leucine zipper segment (Alm-LeuZ), the association number of alamethicin was effectively controlled to produce a single predominant channel open state. The assembly was estimated to be a tetramer, by comparison of the channel conductance with that of the template-assembled Alm LeuZ tetramer, which was prepared by the conjugation of a maleimide functionalized peptide template with cysteine-derivatized Alm-LeuZ segments. Employment of an extramembrane segment of a random conformation provided higher levels of channel conductance. The result exemplified the possibility of channel current control by a conformational switch of the extramembrane segments. PMID- 11734011 TI - Refinement of local and long-range structural order in theophylline-binding RNA using (13)C-(1)H residual dipolar couplings and restrained molecular dynamics. AB - 13C-(1)H residual dipolar couplings (RDC) have been measured for the bases and sugars in the theophylline-binding RNA aptamer, dissolved in filamentous phage medium, and used to investigate the long-range structural and dynamic behavior of the molecule in the solution state. The orientation dependent RDC provide additional restraints to further refine the overall structure of the RNA theophylline complex, whose long-range order was poorly defined in the NOE-based structural ensemble. Structure refinement using RDC normally assumes that molecular alignment can be characterized by a single tensor and that the molecule is essentially rigid. To address the validity of this assumption for the complex of interest, we have analyzed distinct domains of the RNA molecule separately, so that local structure and alignment tensors experienced by each region are independently determined. Alignment tensors for the stem regions of the molecule were allowed to float freely during a restrained molecular dynamics structure refinement protocol and found to converge to similar magnitudes. During the second stage of the calculation, a single alignment tensor was thus applied for the whole molecule and an average molecular conformation satisfying all experimental data was determined. Semirigid-body molecular dynamics calculations were used to reorient the refined helical regions to a relative orientation consistent with this alignment tensor, allowing determination of the global conformation of the molecule. Simultaneously, the local structure of the theophylline-binding core of the molecule was refined under the influence of this common tensor. The final ensemble has an average pairwise root mean square deviation of 1.50 +/- 0.19 A taken over all heavy atoms, compared to 3.5 +/- 1.1 A for the ensemble determined without residual dipolar coupling. This study illustrates the importance of considering both the local and long-range nature of RDC when applying these restraints to structure refinements of nucleic acids. PMID- 11734012 TI - A novel nitroimidazole compound formed during the reaction of peroxynitrite with 2',3',5'-tri-O-acetyl-guanosine. AB - Peroxynitrite reacts with 2',3',5'-tri-O-acetyl-guanosine to yield a novel compound identified as 1-(2,3,5-tri-O-acetyl-beta-D-erythro-pentofuranosyl)-5 guanidino-4-nitroimidazole (6). This characterization was achieved using a combination of UV/vis spectroscopy and ESI-MS. Additionally, 1-(beta-D-erythro pentofuranosyl)-5-guanidino-4-nitroimidazole (6a) was synthesized by an independent route, characterized by UV/vis spectroscopy, ESI-MS, and (1)H- and (13)C NMR, and shown to be identical to deacetylated 6. This product is extremely stable in aqueous solution at both pH extremes and is formed in significant yields. These characteristics suggest that this lesion may be useful as a specific biomarker of peroxynitrite-induced DNA damage. We also observed formation of 2',3',5'-tri-O-acetyl-8-nitroguanosine (2',3',5'-tri-O-acetyl-8 NO(2)()Guo), 2-amino-5-[(2,3,5-tri-O-acetyl-beta-D-erythro-pentofuranosyl)amino] 4H-imidazol-4-one (2',3',5'-tri-O-acetyl-Iz), and the peroxynitrite-induced oxidation products of 2',3',5'-tri-O-acetyl-8-oxoGuo. The formation of 6 and 2',3',5'-tri-O-acetyl-8-NO(2)()Guo was rationalized by a mechanism invoking formation of the guanine radical. PMID- 11734013 TI - Artificial ion channels showing rectified current behavior. AB - Voltage-dependent artificial ion channels 3 and 4 were synthesized. Two cholic acid derivatives were connected through a m-xylylene dicarbamate unit at 3 hydroxyl groups. Asymmetries were introduced by terminal hydrophilic groups, carboxylic acid and phosphoric acid for 3 and hydroxyl and carboxylic acid for 4. Under basic conditions, these headgroups in 3 and 4 are expected to be dissociate into -1/-2 (pH 8.2) and 0/-1 (pH 7.2), respectively. Single ion channel properties were examined by a planar bilayer lipid membrane method under symmetrical 500 mM KCl at pH 8.2 or 7.2. When 3 and 4 were introduced into the bilayer membrane under application of positive voltage (a positive-shift method), the current values at positive applied voltage were larger than the corresponding ones at the negative applied voltage. The current-voltage plots were fitted by curves through a zero point to show clear rectification properties. The direction of rectification could be controlled by positive- or negative-shift methods. Vectorial alignment of terminal headgroup charges by the voltage-shift incorporation is essential for giving voltage-dependent rectified ion channels. PMID- 11734014 TI - Stability, reactivity, solution, and solid-state structure of halomethylzinc alkoxides. AB - In this paper, we report our findings regarding the development of a Lewis acid catalyzed cyclopropanation of allylic alcohols with bis(iodomethyl)zinc. Iodomethylzinc alkoxides can be formed by treatment of an alcohol with bis(iodomethyl)zinc. These species are not prone to undergo cyclopropanation at low temperature but the addition of a Lewis acid in catalytic amounts induces the cyclopropanation reaction. Using this procedure, we demonstrated that the Lewis acid-catalyzed pathway significantly overwhelms the uncatalyzed one. This paper describes fundamental issues regarding the preparation and stability of halomethyl zinc alkoxides in solution as well as their aggregation state in solution and solid-state structures. Furthermore, the competition reaction between the inter- vs intramolecular cyclopropanation will be studied. Finally, we will discuss the possible activation pathways to explain the Lewis acid activation of halomethylzinc alkoxides. These findings provided new insights on the reactivity of ROZnCH(2)I and established the groundwork for the elaboration of an enantioselective version of the reaction. PMID- 11734015 TI - Catalytic asymmetric cyclopropanation of allylic alcohols with titanium TADDOLate: scope of the cyclopropanation reaction. AB - A substoichiometric amount of titanium-TADDOLate complex was effective at catalyzing the cyclopropanation reaction of allylic alcohols in the presence 1 equiv of bis(iodomethyl)zinc. After initial optimization of the catalyst structure, excellent yields and enantiomeric ratios were obtained for 3-aryl- or 3-heteroaryl-substituted allylic alcohols (up to 97:3). Alkyl-substituted allylic alcohols gave modest yields and enantiomeric ratios (up to 87:13) but these compare favorably with those observed with other substoichiometric chiral ligands. The full synthetic scope of the reaction is presented in this paper. PMID- 11734016 TI - Synchronized propagation mechanism for crystalline-state polymerization of p xylylenediammonium disorbate. AB - We describe the results of the topochemical polymerization of p xylylenediammonium disorbate as the bifunctional monomer in the crystalline state via a radical chain polymerization mechanism. The structure and properties of the resulting double-stranded polymer are investigated by X-ray crystal structure and thermal analyses as well as IR spectroscopy. The stereochemical structure of the polymer is confirmed by NMR spectroscopy after the solid-state transformation of the polymer side chain to give the single-stranded chains that are soluble in methanol. We propose a model for the reaction mechanism to provide a ladder polymer in the crystalline state on the basis of the results of single-crystal structure analysis and the observation of the alpha-methyl and alpha-carboxylate propagating radicals by ESR spectroscopy. The two reaction parts of the monomer are synchronized to simultaneously react with the conformational change in a sorbate moiety that is transformed into another sorbate through the diammonium part as the linker. PMID- 11734017 TI - New efficient calixarene amide ionophores for the selective removal of strontium ion from nuclear waste: synthesis, complexation, and extraction properties. AB - Three novel lower rim hexamide derivatives 5(6), 7(6), and 9(6) of p hydroxycalix[6]arene and four octamides 5(8), 7(8)-9(8) derived from the corresponding p-hydroxycalix[8]arene were synthesized, and their potential as extractants in radioactive waste treatment was evaluated, in comparison with upper rim analogues 12(6) and 12(8) and other existing selective neutral ionophores currently used in radioactive waste treatment. Extraction of alkali and alkaline earth metal picrates from water to dichloromethane, and of the corresponding nitrates from acidic water solution simulating radioactive waste, to 2-nitrophenyl hexyl ether (NPHE), showed that the lower rim amides extract divalent cations much better than monovalent ones. The upper rim hexa-12(6) and octamide 12(8) are very inefficient ligands, hardly extracting any cation. In all cases, p-alkoxy octamides are more efficient and selective extractants than the corresponding hexamides. In the case of simulated waste solutions, the distribution coefficients for strontium removal by octamides (6.5 < D(Sr) < 30) are much higher than the corresponding value (D(Sr)) found for dicyclohexyl-18 crown-6 (DC18C6), and the same applies for the strontium/sodium selectivity, which is 6500 < D(Sr)/D(Na) < 30 000 for octamides and 47 for DC18C6. ESI-MS, UV vis, and X-ray crystal structure studies give consistent results and indicate the formation of 2:1 (cation/ligand) strontium complexes for all octamides tested. Stability constants were determined in homogeneous methanol solution for alkali metal (log beta(11) < or = 2), calcium (4.3 < or = log beta(11) < or = 6.0; 9.4 < or = log beta(21) < or = 12.0), and strontium (5.6 < or = log beta(11) < or = 12.3) ions using a UV-vis competition method with 1-(2-pyridylazo)-2-naphthol (PAN). They confirm the high efficiency and high divalent/monovalent selectivity found in metal ion extraction experiments for the new octamide ligands. Evidence for a positive cooperative effect between the two metal ion binding sites was obtained in the case of the Ca(2+) complex of octamide 1(8). PMID- 11734018 TI - gem-Diacetates as carbonyl surrogates for asymmetric synthesis. Total syntheses of sphingofungins E and F. AB - The equivalent of an asymmetric addition to a carbonyl group with a stabilized anion is accomplished by discriminating between the enantiotopic C-O single bonds of a gem-diacetate. In this way, enantioselective total syntheses of two antifugal agents, sphingofungins E and F, have been accomplished. The synthetic strategy is based on a series of catalytic processes whereby all of the chiral centers are created with high stereoselectivities. The first two stereocenters are introduced by an asymmetric allylic alkylation reaction of gem-diacetate 9 with azlactone 10. The complex of Pd(0) and ligand 14 efficiently catalyzes this key reaction, which differentiates both the enantiotopic leaving groups of a gem diacetate and enantiotopic faces of the enolate of an azlactone in high enantiomeric excess and diastereomeric excess. From these two stereocenters, the configurations of the remaining two centers are set by a diastereoselective Os(VIII)-catalyzed dihydroxylation reaction with excellent stereocontrol. The trans-alkene is established by Cr(II)-mediated olefination, and a subsequent B alkyl Suzuki coupling reaction conjoins the polar head unit and the nonpolar, 13 carbon lipid tail. The efficiency of our strategy is illustrated by the completion of syntheses of sphingofungins F and E in 15 and 17 steps, and in 17% and 5% overall yields, respectively. PMID- 11734019 TI - Palladium-catalyzed intramolecular C-O bond formation. AB - A number of oxygen heterocycles were synthesized using the palladium-catalyzed intramolecular etherification of aryl halides by employing di-tert butylphosphinobiaryl ligands. The reaction proceeds under mild conditions using weak bases such as Cs(2)CO(3) or K(3)PO(4). A variety of functional groups are tolerated in the reaction, and enantioenriched alcohols can be coupled without erosion of optical purity. The mildness of the reaction conditions allows for the use of polyfunctionalized substrates. This method was used as the key step in the synthesis of MKC-242, an antidepressant currently in clinical trials. The synthesis of MKC-242 was achieved in 40% overall yield from commercially available sesamol and acrylonitrile. PMID- 11734020 TI - Electronic absorption spectroscopy of cobalt ions in diluted magnetic semiconductor quantum dots: demonstration of an isocrystalline core/shell synthetic method. AB - This paper reports the application of ligand-field electronic absorption spectroscopy to probe Co(2+) dopant ions in diluted magnetic semiconductor quantum dots. It is found that standard inverted micelle coprecipitation methods for preparing Co(2+)-doped CdS (Co(2+):CdS) quantum dots yield dopant ions predominantly bound to the nanocrystal surfaces. These Co(2+):CdS nanocrystals are unstable with respect to solvation of surface-bound Co(2+), and time dependent absorption measurements allow identification of two transient surface bound intermediates involving solvent-cobalt coordination. Comparison with Co(2+):ZnS quantum dots prepared by the same methods, which show nearly isotropic dopant distribution, indicates that the large mismatch between the ionic radii of Co(2+) (0.74 A) and Cd(2+) (0.97 A) is responsible for exclusion of Co(2+) ions during CdS nanocrystal growth. An isocrystalline core/shell preparative method is developed that allows synthesis of internally doped Co(2+):CdS quantum dots through encapsulation of surface-bound ions beneath additional layers of CdS. PMID- 11734021 TI - Octahedral Fe(II) and Ru(II) complexes based on a new bis 1,10-phenanthroline ligand that imposes a well defined axis. AB - A bis-chelating ligand (L1), made of two 7-(p-anisyl)-1,10-phenanthroline (phen) subunits connected with a p-(CH(2))(2)C(6)H(4)(CH(2))(2) spacer through their 4 positions, has been prepared, using Skraup syntheses and reaction of the anion of 4-methyl-7-anisyl-1,10-phenanthroline with alpha,alpha'-dibromo-p-xylene. Its Fe(II) complex, [FeL1(dmbp)](PF(6))(2), was prepared in one step by reaction of L1 with [Fe(dmbp)(3)](PF(6))(2) (dmbp = 4,4'-dimethyl-2,2'-bipyridine). On the other hand, its Ru(II) complex, [RuL1(dmbp)](PF(6))(2), was prepared in two steps from Ru(CH(3)CN)(4)Cl(2) and L1, followed by reaction with dmbp. X-ray crystal structure analyses show that in the two octahedral complexes, ligand L1 coils around the metal by coordination of the axial and two equatorial positions. It defines a 21 A long axis (O.O distance) running through the central metal and the terminal anisyl substituents. The complexes were also characterized by (1)H NMR, mass spectrometry, cyclic voltammetry, electronic absorption, and, in the case of Ru(II), fluorescence spectroscopy. PMID- 11734022 TI - Transition-metal-promoted reactions of boron hydrides. 17. Titanium-catalyzed decaborane-olefin hydroborations. AB - The titanium-catalyzed hydroboration reactions of decaborane with a variety of terminal olefins have been found to result in the exclusive, high-yield formation of monosubstituted decaborane 6-R-B(10)H(13) products, arising from anti Markovnikov addition of the cage B6-H to the olefin. The titanium-catalyzed reactions are slow, often less than one turnover per hour; however, their high selectivities and yields coupled with the fact that they are simple, one-pot reactions give them significant advantages over the previously reported routes to 6-R-B(10)H(13) compounds. The catalyst also has extended activity with reactions carried out for as long as 13 days, showing little decrease in reactivity, thereby allowing for the production of large amounts of 6-R-B(10)H(13). The titanium-catalyzed reactions of decaborane with the nonconjugated diolefins, 1,5 hexadiene and diallylsilane, were found to give, depending upon reaction conditions and stoichiometries, high yields of either alkenyl-substituted 6 (CH(2)=CH(CH(2))(4))-B(10)H(13) (4) and 6-(CH(2)=CHCH(2)SiMe(2)(CH(2))(3)) B(10)H(13) (5) or linked-cage 6,6'-(CH(2))(6)-(B(10)H(13))(2) (6) and Me(2)Si(6 (CH(2))(3)-B(10)H(13))(2) (7) compounds, respectively. The unique tetra-cage product, Si(6-(CH(2))(3)-B(10)H(13))(4) (8), was obtained by the catalyzed reaction of 4 equiv of decaborane with tetraallylsilane. Sequential use of the titanium catalyst and previously reported platinum catalysts (PtBr(2) or H(2)PtCl(6).6H(2)O with an initiator) provides an efficient pathway to asymmetrically substituted 6-R-9-R'-B(10)H(12) species. The structures of compounds 5, 6, and 8, as well as a platinum derivative, (PSH(+))(2)-commo-Pt [nido-7-Pt-8-(n-C(8)H(17))B(10)H(11)](2)(2-), of 6-(n-octyl)decaborane have been established by single-crystal crystallographic determinations. PMID- 11734024 TI - Involvement of water in carbohydrate-protein binding. AB - The interactions of trimannosides 1 and 2 with Con A were studied to reveal the effects of displacement of well-ordered water molecules on the thermodynamic parameters of protein-ligand complexation. Trisaccharide 2 is a derivative of 1, in which the hydroxyl at C-2 of the central mannose unit is replaced by a hydroxyethyl moiety. Upon binding, this moiety displaces a conserved water molecule present in the Con A binding site. Structural studies by NMR spectroscopy and MD simulations showed that the two compounds have very similar solution conformational properties. MD simulations of the complexes of Con A with 1 and 2 demonstrated that the hydroxyethyl side chain of 2 can establish the same hydrogen bonds in a low energy conformation with the protein binding site as those mediated by the water molecule in the complex of 1 with Con A. Isothermal titration microcalorimetry (ITC) measurements showed that 2 has a more favorable entropy of binding compared to 1. This term, which was expected, arises from the return of the highly ordered water molecule to bulk solution. The favorable entropy term was, however, offset by a relatively large unfavorable enthalpy term. This observation was rationalized by comparing the extent of hydrogen bond and solvation changes during binding. It is proposed that an indirect interaction through a water molecule will provide a larger number of hydrogen bonds in the complex that have higher occupancies than in bulk solution, thereby stabilizing the complex. PMID- 11734023 TI - Photochromic atropisomer generation and conformation determination in a ruthenium bis(bipyridine) phosphonite gamma-cyclodextrin system. AB - Irradiation of rac-[Ru(bpy)(2)(PhP(OMe)(2))(Cl)]Cl (2) at lambda > 460 nm results in the photochromic generation of a new atropisomer and chirality inversion, via rotation of the PhP(OMe)(2) moiety around the Ru-P bond. However, since the energetic barrier to rotation is low resulting in racemization, it was found that the formation of a supramolecular complex between 2 and gamma-cyclodextrin (gamma CDx) facilitated the stabilization of the new atropisomeric conformation. On irradiation the bisignate signals in the circular dichroism spectrum of the 2:gamma-CDx complex were converted to an entirely new and distinct circular dichroism spectrum, as a result of a different spatial orientation of the phenyl electronic transition in the PhP(OMe)(2) moiety (the active circular dichroism spectra were found to arise from different g factor values of the Delta-2:gamma CDx and Lambda-2:gamma-CDx complexes). The new atropisomer formation and subsequent thermally induced interconversion could be further detected by conventional and variable temperature (1)H NMR studies. Determination of the conformation of the new atropisomer was achieved by combining analysis of the changes in the circular dichroism spectra by exciton coupling theory with molecular modeling and DFT calculations. PMID- 11734025 TI - The unexpected and large enhancement of the dipole moment in the 3,4 bis(dimethylamino)-3-cyclobutene-1,2-dione (DMACB) molecule upon crystallization: a new role of the intermolecular CH...O interactions. AB - The molecular dipole moment of the 3,4-bis(dimethylamino)-3-cyclobutene-1,2-dione (DMACB) molecule and its enhancement in the crystal was evaluated by periodic RHF ab initio computations. A discrete boundary partitioning of the electronic density that allows an unambiguous partitioning of the molecular space in the condensed phase was adopted. The resulting molecular dipole in the crystal compares favorably with the experimental value obtained by a multipolar analysis of single-crystal X-ray diffraction data recorded at 20 K, using a fuzzy boundary partitioning of the derived pseudoatom densities. We show that a large and highly significant molecular dipole enhancement may occur upon crystallization, despite the lack of a strongly hydrogen bonded environment in the crystal. The 23 unique C-H...O interactions which are formed upon packing of the DMACB molecule induce an increase in the molecular dipole (over 75%) that is comparable to or greater than that found in systems which are characterized by the stronger O-H...O and N H...O hydrogen bonds. The DMACB molecule constitutes an excellent system for the study of C-H...O interactions in the condensed phase, since no other kind of competing hydrogen bonds is present in its crystal. A simple and qualitative model for the matrix contribution to the DMACB molecular dipole enhancement in the crystal is proposed. The formation of several weak C-H...O bonds is found to yield a small (about 0.2 e) net flux of electronic charge flowing from the hydrogens of the methyl groups to the carbonyl oxygen atoms. Despite the limited increase of the intramolecular charge transfer upon crystallization, a large molecular dipole enhancement occurs because the centroids of the positive and negative induced charges are quite far apart. This work highlights a new and important role of the C-H...O bond, besides those already known in the literature. PMID- 11734026 TI - Effect of metal ion and water coordination on the structure of a gas-phase amino acid. AB - The mode of metal ion and water binding to the amino acid valine is investigated using both theory and experiment. Computations indicate that without water, the structure of valine is nonzwitterionic. Both Li(+) and Na(+) are coordinated to the nitrogen and carbonyl oxygen (NO coordination), whereas K(+) coordinates to both oxygens (OO coordination) of nonzwitterionic valine. The addition of a single water molecule does not significantly affect the relative energies calculated for the cationized valine clusters. Experimentally, the rates of water evaporation from clusters of Val.M(+)(H(2)O)(1), M = Li, Na, and K, are measured using blackbody infrared radiative dissociation. The dissociation rate from the valine complex is compared to water evaporation rates from model complexes of known structure. These results indicate that the metal ion in the lithiated and the sodiated clusters is NO-coordinated to nonzwitterionic valine, while that in the potassiated cluster has OO coordination, in full agreement with theory. The zwitterionic vs nonzwitterionic character of valine in the potassiated cluster cannot be distinguished experimentally. Extensive modeling provides strong support for the validity of inferring structural information from the kinetic data. PMID- 11734027 TI - DFT studies on the copolymerization of alpha-olefins with polar monomers: ethylene-methyl acrylate copolymerization catalyzed by a Pd-based diimine catalyst. AB - Gradient-corrected density functional theory has been used to study the elementary reactions for the copolymerization of ethylene with methyl acrylate catalyzed by Pd-based diimine catalysts, modeled by the generic complex N(wedge)N Pd(n-C(3)H(7))(+), with N(wedge)N = -NHCHCHNH-. The steric effects in the real systems are discussed on the basis of the calculations for the catalyst with N(wedge)N = -NArCRCRNAr-, R = CH(3), and Ar = C(6)H(3)(i-Pr(2)) and the previous calculations on ethylene/propylene polymerization. Considerations have been given to the different possible acrylate complexes, as well as the transition states and the products (agostic complexes and the alternative chelates) for two acrylate insertion paths (1,2 and 2,1). The chelate-opening reactions have also been studied. The results revealed a strong electronic preference for the 2,1 insertion paths, with a barrier that is 4.5 kcal/mol lower than any other studied insertion pathway. In the real systems the 2,1-insertion of acrylate is preferred by 0.5 kcal/mol. The 2,1-insertion barrier calculated for the real system of 12.4 kcal/mol is in very good agreement with the experimental value of 12.1 kcal/mol. The six-member chelate is the most stable insertion product with an energy that is 21 kcal/mol lower than the kinetic insertion product. The reactions of the chelate opening by ethylene that start from the lowest energy complexes have the lowest barrier for the four-member ring (23 kcal/mol) and the highest for the six member structure (30.4 kcal/mol). The high barrier for the opening of the six member chelate suggests the possibility of a two-step chelate-opening mechanism. The internal barriers for the chelate-opening reactions starting from the higher energy complexes are lower then the one-step reaction that starts from the preferred complex and comparable to those of the ethylene insertion into the Pd alkyl bond. While the chelate opening by a subsequent acrylate insertion seems to be facile for the generic catalyst, steric effects in the real catalyst are likely to decrease the acrylate pi-complexation energies and increase the insertion barriers to the extent where such a reaction becomes unfeasible. PMID- 11734028 TI - Influence of sodium ions on the dynamics and structure of single-stranded DNA oligomers: a molecular dynamics study. AB - The effects of sodium counterion presence and chain length on the structure and dynamics of single DNA strands of polythymidylate were studied by means of molecular dynamics simulations. The importance of the base-base stacking phenomenon increases with the chain length and partially reduces the flexibility of the strand. Sodium ions directly interact with the phosphate groups and keto oxygens of the thymine bases, complexes showing lifetimes below 400 ps. Simultaneous phosphate and keto complexes were observed for one of the sodium ions with lifetimes around 1 ns. The implications of such complexes in the folding process experienced by the strand are considered. Structurally, cation inner- and outer-sphere complexes were observed in the coordination of phosphate groups. For the inner-sphere complexes, the structural information retrieved from the simulations is in very good agreement with experimental data. The diffusion properties of the sodium ions also reflect both types of coordination modes. PMID- 11734029 TI - The nature of improper, blue-shifting hydrogen bonding verified experimentally. AB - In the infrared spectra of solutions in liquid argon of dimethyl ether ((CH(3))(2)O) and fluoroform (HCF(3)), bands due to a 1:1 complex between these monomers have been observed. The C-H stretch of the HCF(3) moiety in the complex appears 17.7 cm(-1) above that in the monomer, and its intensity decreases by a factor of 11(2). These characteristics situate the interaction between the monomers in the realm of improper, blue-shifting hydrogen bonding. The complexation shifts the C-F stretches downward by some 9 cm(-1), while the C-H stretches in (CH(3))(2)O are shifted upward by 9-15 cm(-1), and the C-O stretches are shifted downward by 5 cm(-1). These shifts are in very good agreement with those calculated by means of correlated ab initio methods, and this validates a two-step mechanism for improper, blue-shifting hydrogen bonding. In the first step, the electron density is transferred from the oxygen lone electron pairs of the proton acceptor ((CH(3))(2)O) to fluorine lone electron pairs of the proton donor (CHF(3)) which yields elongation of all CF bonds. Elongation of CF bonds is followed (in the second step) by structural reorganization of the CHF(3) moiety, which leads to the contraction of the CH bond. It is thus clearly demonstrated that not only the spectral manifestation of H-bonding and improper H-bonding but also their nature differ. PMID- 11734030 TI - A theoretical study of steric and electronic effects in the rhodium-catalyzed carbonylation reactions. AB - We present a QM and QM/MM study of steric and electronic effects in the main steps of Rh-catalyzed carbonylation reactions. All the considered systems adopt a square-planar geometry prior to CH(3)I oxidative addition. As regards the octahedral complexes after CH(3)I oxidative addition, a comparison between the various models indicates that the energy gain due to the CH(3)I oxidative addition is reduced by the steric pressure of the substituents on the ligand. The substantially similar results obtained with the QM/MM and QM models indicate that electronic effects are not particularly relevant in determining the energetic of oxidative addition. As regards the P,P-Ph octahedral complex, the geometries in which the CO group is trans to the added CH(3) group, or trans to one of the P atoms, are of similar energy. A comparison between the various models indicates that the energy barrier of the CO insertion reaction is lowered by the presence of substituents on the chelating ligands. This effect is related to a relief of the steric pressure on the complex as the systems move from a six-coordinated octahedral geometry toward a five-coordinated square-pyramidal geometry. The energy barrier calculated for the P,S-Ph system is in rather good agreement with the experimental value, whereas that of the P,P-Ph system is somewhat underestimated. Inclusion of solvent effects with a continuum model leads to a slightly better agreement. The thermodynamic products adopt a square-pyramidal geometry with the COCH(3) group in the apical position. PMID- 11734031 TI - Interplay of the main chain, chiral side chains, and solvent in conformational transitions: poly([(R)-3,7-dimethyloctyl]-[(S)-3-methylpentyl]silylene). AB - Light scattering, sedimentation equilibrium, viscosity, circular dichroism (CD), and UV absorption (UV) measurements were made on dilute solutions of poly([(R) 3,7-dimethyloctyl]-[(S)-3-methylpentyl]silylene)(PRS) as functions of molecular weight. From light scattering and viscosity data, PRS is found to be a very stiff polymer of persistence length q as large as 103 nm at 25 degrees C, essentially a 7(3) helix found in the solid state; q increases only gradually with lowering temperature between -15 and 25 degrees C. The CD data show that PRS undergoes a conformational transition around 3 degrees C in isooctane (transition temperature T(c)). The CD signal is largely positive at low temperatures, passes through zero at T(c), and becomes largely negative at higher temperatures; T(c) is independent of sample's chain length N. This is a highly cooperative helix (M)-to-helix (P) transition depending remarkably on N, as PRS is substantially rodlike. The CD data are converted to the fraction f(P) of P helix as a function of N and analyzed successfully by a statistical mechanical theory based on a helix reversal model, where a polymer chain consists of M and P helices intervened by helix reversals, with the result that the free energy difference DeltaG(h) between P and M shows a temperature dependence similar to that of 2f(P) - 1, whereas the helix reversal energy is substantially constant at 1.2 x 10(4) J mol( 1); the latter value means that the helix reversal occurs only once in 100 Si units or less. This DeltaG(h) change and solvent dependence of T(c) are explained by a double-well potential for the rotation about Si-Si bonds, which incorporates into DeltaG(h) the solvent interactions with the helical grooves of side chains surrounding the main chain. Detailed features of UV absorption spectra at different temperature and molecular weights are also presented. PMID- 11734032 TI - Triradical thermochemistry from collision-induced dissociation threshold energy measurements. The heat of formation of 1,3,5-trimethylenebenzene. AB - A method for measuring the heats of formation of triradicals using energy resolved collision-induced dissociation (CID) of chloro-substituted biradical negative ions is described. This method is applied to the determination of the heat of formation of 1,3,5-trimethylenebenzene, which was generated by CID of the 5-chloromethyl-m-xylylene ion. The measured CID threshold energy for chloride loss (0.83 +/- 0.07 eV) is combined with the electron affinity of the 5 chloromethyl-m-xylylene biradical (1.120 +/- 0.059 eV) to give a heat of formation of the triradical of 111.0 +/- 4.1 kcal/mol that agrees with the bond additivity value of 109.3 +/- 2.1 kcal/mol. The measured heat of formation indicates a third C-H bond dissociation energy (BDE) in 1,3,5-trimethylbenzene of 88.2 +/- 5.0 kcal/mol, indistinguishable from the C-H BDE in toluene or the first or second C-H BDEs in m-xylene. The results are in agreement with the predictions made on the basis of simple qualitative and high-level molecular orbital theories that predict negligible interaction between the unpaired electrons in the high spin triradical. PMID- 11734033 TI - Effects of amino acid phi,psi propensities and secondary structure interactions in modulating H alpha chemical shifts in peptide and protein beta-sheet. AB - H alpha chemical shifts are often used as indicators of secondary structure formation in protein structural analysis and peptide folding studies. On the basis of NMR analysis of model beta-sheet and alpha-helical peptides, together with a statistical analysis of protein structures for which NMR data are available, we show that although the gross pattern of H alpha chemical shifts reflects backbone torsion angles, longer range effects from distant amino acids are the dominant factor determining experimental chemical shifts in beta-sheets of peptides and proteins. These show context-dependent variations that aid structural assignment and highlight anomalous shifts that may be of structural significance and provide insights into beta-sheet stability. PMID- 11734034 TI - Conformation and protonation of 2,2'-bipyridine and 4,4'-bipyridine in acidic aqueous media and acidic ZSM-5 zeolites: a Raman scattering study. AB - In situ FT-Raman scattering spectroscopy was used to monitor the sorption kinetics of 2,2'- and 4,4'-bipyridine in acidic ZSM-5 zeolites. The data processing of all the Raman spectra was applied to extract the characteristic Raman spectra of occluded species and respective Raman contribution generated from many spectral data which resolves spectrum of mixture into pure component spectra without any prior information. The assignment of the extracted spectra was performed according to careful comparison with corresponding spectra extracted from a set of Raman spectra recorded during the protonation of 2,2'- or 4,4'-bipyridine (bpy) in hydrochloric acid aqueous solutions. The data processing of the Raman spectra recorded during the slow sorption of 4,4'-bpy in acidic H(n)ZSM-5 (n = 3, 6) zeolites provides specific Raman spectrum of N,N' diprotonated dication 4,4'-bpyH(2)(2+) as unique species generated in the void space of acidic ZSM-5 zeolites. No evidence of Lewis acid sites was found during the sorption of 4,4'-bpy by Raman scattering spectroscopy. The data processing of the Raman spectra recorded during the slow sorption of 2,2'-bpy in acidic H(n)ZSM 5 (n = 3, 6) zeolites provides specific Raman spectrum of trans-N-monoprotonated cation 2,2'-bpyH+ as major species generated in the void space of acidic ZSM-5 zeolites at loading corresponding to 1 mol per unit cell. The trans/cis interconversion occurs at higher loading even after the complete uptake of the sorbate and indicates some rearrangement in the void space over a long time. The cations were found to be located in straight channels in the vicinity of the intersection with the zigzag channel of the porous materials with the expected conformations deduced from ab initio calculations. However, the motions of occluded species within the channel of ZSM-5 are hindered but remain in the range of the isotropic limit of a liquid at room temperature. PMID- 11734035 TI - Electroosmotic flow in template-prepared carbon nanotube membranes. AB - Carbon nanotube membranes (CNMs) were prepared by doing chemical vapor deposition of carbon within the pores of a microporous alumina template. Electroosmotic flow (EOF) was driven across the CNMs by allowing the membrane to separate two electrolyte solutions and using an electrode in each solution to pass a constant ionic current through the nanotubes. EOF was investigated by measuring the flux of a probe molecule (phenol) across the CNM. The as-synthesized CNMs have anionic surface charge, and EOF is in the direction of cation migration across the membrane. Measurements of the rate of EOF as a function of applied transmembrane current provided the zeta potential. The effect of pH on zeta provided the pK(a) for the surface acidic sites responsible for this anionic charge; the acidic-site density was also determined. An electrochemical derivatization method was used to attach carboxylate groups to the nanotube walls; this enhances the anionic surface charge density, resulting in a corresponding increase in the EOF rate. Electrochemical derivatization was also used to attach cationic ammonium sites to the nanotube walls to yield CNMs that show EOF in the opposite direction of the as-synthesized or carboxylated membranes. PMID- 11734036 TI - Morphology of three lyotropic liquid crystalline biological NMR media studied by translational diffusion anisotropy. AB - The morphologies of three dilute liquid crystalline phases, which are widely used for biological NMR spectroscopy, are investigated by the study of tracer self diffusion. The aqueous liquid crystalline media investigated include the common phospholipid bicelle medium, a phase consisting of a mixture of pentaethyleneglycol mono dodecyl ether and hexanol, and a medium containing cetylpyridinium bromide and hexanol. Threonine and water were used as tracer molecules for probing the aqueous environment, and tetramethylsilane (TMS) was for probing the lipophilic environment. Pulsed field gradient NMR was used to measure tracer self-diffusion rates in three orthogonal directions. Although results for the water-soluble tracers in bicelle media do not contradict the widely accepted disk-shaped bicelle model, the high TMS diffusion rate observed in the bilayer plane requires extensive transient edge-to-edge contacts of such disks. This morphology is essentially that of a heavily perforated lamellar bilayer phase and explains why this medium remains liquid crystalline well below the Onsager limit for disk-shaped nematogens. Below 25 degrees C, a bicelle mixture consisting of dimyristoyl phosphatidyl choline and dihexanoyl phosphatidyl choline remains isotropic, but tracer diffusion obstruction indicates that the particles are significantly oblate. The diffusion anisotropy in the penta(ethyleneglycol) mono dodecyl ether liquid crystals confirms the previously proposed alpha-lamellar phase. However, weak inhibition of aqueous phase self-diffusion in the z direction points to the presence of bridge- or caplike obstructions, and the bilayers appear slightly permeable to water. If the previously proposed concentric cylinder superstructure of bilayers applies, the diffusion data indicate that the most outer cylinder must have a diameter greater than 50 microm. The tracer self-diffusion data for the cetylpyridinium bromide/hexanol medium is only compatible with a planar alpha-lamellar phase, with its local director orthogonal to the magnetic field, and a very large domain size over which the director remains parallel. PMID- 11734037 TI - Microwave spectra and molecular structures of (Z)-pent-2-en-4-ynenitrile and maleonitrile. AB - Accurate equilibrium structures have been determined for (Z)-pent-2-en-4 ynenitrile (8) and maleonitrile (9) by combining microwave spectroscopy data and ab initio quantum chemistry calculations. The microwave spectra of 10 isotopomers of 8 and 5 isotopomers of 9 were obtained using a pulsed nozzle Fourier transform microwave spectrometer. The ground-state rotational constants were adjusted for vibration-rotation interaction effects calculated from force fields obtained from ab initio calculations. The resultant equilibrium rotational constants were used to determine structures that are in very good agreement with those obtained from high-level ab initio calculations (CCSD(T)/cc-pVTZ). The geometric parameters in 8 and 9 are very similar; they also do not differ significantly from the all carbon analogue, (Z)-hex-3-ene-1,5-diyne (7), the parent molecule for the Bergman cyclization. A small deviation from linearity about the alkyne and cyano linkages is observed for 7-9 and several related species where accurate equilibrium parameters are available. The data on 7-9 should be of interest to radioastronomy and may provide insights on the formation and interstellar chemistry of unsaturated species such as the cyanopolyynes. PMID- 11734039 TI - Remarkable stability of hairpins containing 2',5'-linked RNA loops. AB - We report here the results of a comparative study of hairpin loops that differ in the connectivity of phosphodiester linkages (3',5'- versus 2',5'-linkages). In addition, we have studied the effect of changing the stem composition on the thermodynamic stability of hairpin loops. Specifically, we constructed hairpins containing one of six stem duplex combinations, i.e., DNA:DNA ("DD"), RNA:RNA ("RR"), DNA:RNA ("DR"), 2',5'-RNA:RNA ("RR"), 2',5'-RNA:DNA ("RD"), and 2',5' RNA:2',5'-RNA ("RR"), and one of three tetraloop compositions, i.e., 2',5'-RNA ("R"), RNA ("R"), and DNA ("D"). All hairpins contained the conserved and well studied loop sequence 5'-...C(UUCG)G...-3' [Cheong et al. Nature 1990, 346, 680 682]. We show that the 2',5'-linked loop C(UUCG)G, i.e.,...C(3'p5')U(2'p5')U(2'p5')C(2'p5')G(2'p5')G(3'p5')..., like its "normal" RNA counterpart, forms an unusually stable tetraloop structure. We also show that the stability imparted by 2',5'-RNA loops is dependent on base sequence, a property that is shared with the regioisomeric 3',5'-RNA loops. Remarkably, we find that the stability of the UUCG tetraloop is virtually independent of the hairpin stem composition (DD, RR, RR, etc.), whereas the native RNA tetraloop exerts extra stability only when the stem is duplex RNA (R:R). As a result, the relative stabilities of hairpins with a 2',5'-linked tetraloop, e.g. ggac(UUCG)gtcc (T(m) = 61.4 degrees C), are often superior to those with RNA tetraloops, e.g. ggac(UUCG)gtcc (T(m) = 54.6 degrees C). In fact, it has been possible to observe the formation of a 2',5'-RNA:DNA hybrid duplex by linking the hybrid's strands to a (UUCG) loop. These duplexes (RD), which are not stable enough to form in an intermolecular complex [Wasner et al. Biochemistry 1998, 37, 7478-7486], were stable at room temperature (T(m) approximately 50 degrees C). Thus, 2',5'-loops have potentially important implications in the study of nucleic acid complexes where structural data are not yet available. Furthermore, they may be particularly useful as structural motifs for synthetic ribozymes and nucleic acid "aptamers". PMID- 11734038 TI - Structure of a copper-mediated base pair in DNA. AB - Stable and selective DNA base pairing by metal coordination was recently demonstrated with nucleotides containing complementary pyridine-2,6-dicarboxylate (Dipic) and pyridine (Py) bases (Meggers, E.; Holland, P. L.; Tolman; W. B.; Romesberg, F. E.; Schultz, P. G. J. Am. Chem. Soc. 2000, 122, 10714-10715). To understand the structural consequences of introducing this novel base pair into DNA we have solved the crystal structure of a duplex containing the metallo-base pair. The structure shows that the bases pair as designed, but in a Z-DNA conformation. The structure also provides a structural explanation for the B- to Z-DNA transition in this duplex. Further solution studies demonstrate that the metallo-base pair is compatible with Z- or B-DNA conformations, depending on the duplex sequence. PMID- 11734040 TI - Unique [(1)infinityNi(8)Bi(8)S] metallic wires in a novel quasi-1D compound. Synthesis, crystal and electronic structure, and properties of Ni(8)Bi(8)SI. AB - A new quasi-one-dimensional compound Ni(8)Bi(8)SI has been synthesized and its crystal structure determined from single-crystal X-ray diffraction data. The structure of Ni(8)Bi(8)SI consists of [(1)infinityNi(8)Bi(8)S] columns separated by iodine atoms. Conductivity and magnetic susceptibility measurements (down to 4.2 K) show that Ni(8)Bi(8)SI is a one-dimensional metal and exhibits Pauli paramagnetic properties. These observations are in good agreement with the results from electronic structure calculations. An analysis of the chemical bonding employing difference electron charge density maps reveals strong multicenter Ni-Bi bonds and pair Ni-S interactions within the [(1)infinityNi(8)Bi(8)S] columns. Only electrostatic interactions are inferred between the columns and iodine atoms. PMID- 11734041 TI - Lattice-Boltzmann simulations of self-assembly of a binary water-surfactant system into ordered bicontinuous cubic and lamellar phases. AB - We used our recently developed mesoscale amphiphilic lattice-Boltzmann method (Nekovee, M.; Coveney, P. V.; Chen, H.; Boghosian, B. M. Phys. Rev. E 2000, 62, 8282-8894) to investigate the dynamics of self-assembly of the bicontinuous cubic phase in a binary water-surfactant system, and the transition from the lamellar structure to a bicontinuous cubic phase. Our study provides insight into how such structures emerge as a result of competing molecular interactions between water and amphiphiles and among amphiphilic molecules themselves, and represents the first application of any lattice-Boltzmann model to amphiphilic systems in three dimensions. PMID- 11734042 TI - Molecular dynamics simulation of a lipid diamond cubic phase. AB - This paper presents the first atomistic simulation of a cubic membrane phase. Using the molecular dynamics simulation technique both the global and the local organization of glycerolmonoolein molecules inside the diamond cubic phase are studied. Multinanosecond simulations reveal that the center of the cubic bilayer remains close to the infinite periodic minimal surface that describes the diamond geometry. We further show that the equilibrium structure of the surfactant molecules inside the cubic phase is very similar to their structure inside a simulated lamellar bilayer. The small differences arise from the packing constraints of the surfactants within the cubic phase which has an area per surfactant that increases toward the bilayer center. PMID- 11734043 TI - FeMo cofactor of nitrogenase: a density functional study of states M(N), M(OX), M(R), and M(I). AB - The M(N) S = (3)/(2) resting state of the FeMo cofactor of nitrogenase has been proposed to have metal-ion valencies of either Mo(4+)6Fe(2+)Fe(3+) (derived from metal hyperfine interactions) or Mo(4+)4Fe(2+)3Fe(3+) (from Mossbauer isomer shifts). Spin-polarized broken-symmetry (BS) density functional theory (DFT) calculations have been undertaken to determine which oxidation level best represents the M(N) state and to provide a framework for understanding its energetics and spectroscopy. For the Mo(4+)6Fe(2+)Fe(3+) oxidation state, the spin coupling pattern for several spin state alignments compatible with S = (3)/(2) were generated and assessed by energy and geometric criteria. The most likely BS spin state is composed of a Mo3Fe cluster with spin S(a) = 2 antiferromagnetically coupled to a 4Fe' cluster with spin S(b) = (7)/(2). This state has a low DFT energy for the isolated FeMoco cluster and the lowest energy when the interaction with the protein and solvent environment is included. This spin state also displays calculated metal hyperfine and Mossbauer isomer shifts compatible with experiment, and optimized geometries that are in excellent agreement with the protein X-ray data. Our best model for the actual spin-coupled state within FeMoco alters this BS state by a slight canting of spins and is analogous in several respects to that found in the 8Fe P-cluster in the same protein. The spin-up and spin-down components of the LUMO contain atomic contributions from Mo(4+) and the homocitrate and from the central prismane Fe sites and muS(2) atoms, respectively. This qualitative picture of the accepting orbitals for M(N) is consistent with observations from Mossbauer spectra of the one-electron reduced states. Similar calculations for the Mo(4+)4Fe(2+)3Fe(3+) oxidation state yield results that are in poorer agreement with experiment. Using the Mo(4+)6Fe(2+)Fe(3+) oxidation level as the most plausible resting state, the geometric, electronic and energetic properties of the one-electron redox transition to the oxidized state, M(OX), catalytically observed M(R) and radiolytically reduced M(I) states have also been explored. PMID- 11734044 TI - Total synthesis of teicoplanin aglycon. PMID- 11734045 TI - Topographical imaging of an intermembrane junction by combined fluorescence interference and energy transfer microscopies. PMID- 11734046 TI - Octa(aminophenyl)silsesquioxane as a nanoconstruction site. PMID- 11734047 TI - Molecular gain in a thiotolane system. PMID- 11734048 TI - Generation of new enzyme inhibitors using imprinted binding sites: the anti idiotypic approach, a step toward the next generation of molecular imprinting. PMID- 11734049 TI - First unequivocal observation of the whole bell-shaped energy gap law in intramolecular charge separation from S(2) excited state of directly linked porphyrin-imide dyads and its solvent-polarity dependencies. PMID- 11734050 TI - Redox-controlled orthogonal assembly of charged nanostructures. PMID- 11734051 TI - Total synthesis of (+)-zampanolide. PMID- 11734052 TI - Formation of protein-protein complexes in vacuo. PMID- 11734053 TI - The prime reactive intermediate in the iron(III) porphyrin complex catalyzed oxidation reactions by tert-butyl hydroperoxide. PMID- 11734054 TI - Total synthesis of (+)-ambruticin S. PMID- 11734055 TI - Low-temperature hydrogenation of triplet carbenes and diradicaloid biscarbenes- electronic state selectivity. PMID- 11734056 TI - Cyclotetrasilenyl: the first isolable silyl radical. PMID- 11734058 TI - Wolbachia: evolutionary novelty in a rickettsial bacteria. AB - BACKGROUND: Although closely related, the alpha-proteobacteria Wolbachia and the Rickettsiaceae (Rickettsia and Ehrlichia), employ different evolutionary life history strategies. Wolbachia are obligate endocellular symbionts that infect an extraordinary host range and, in contrast to the infectious and pathogenic Rickettsia and Ehrlichia, profoundly influence host reproductive biology. RESULTS: Phylogenies of the Rickettsia, Ehrlichia, and Wolbachia were independently inferred from 16S rDNA sequences and GroEL amino acid sequences. Topologies inferred from both sets of sequence data were consistent with one another, and both indicate the genus Wolbachia shared a common ancestor most recently with Ehrlichia. These two genera are a sister group to the genus Rickettsia. Mapping biological properties onto this phylogeny reveals that manipulation of host reproduction, characteristic of Wolbachia strains, is a derived characteristic. This evolutionary novelty is accompanied by the loss of the ability to infect vertebrate hosts. CONCLUSIONS: Because of the contrasting transmission strategies employed by each, Wolbachia is expected to maximize efficiency of vertical transmission, while Ehrlichia and Rickettsia will optimize horizontal transfer of infection. Wolbachia manipulation of host reproduction could thus be viewed as strategy employed by this bacterium to foster its own propagation via vertical transmission. PMID- 11734059 TI - Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains. AB - BACKGROUND: Despite the medical importance of trichomoniasis, little is known about the genetic relatedness of Trichomonas vaginalis strains with similar biological characteristics. Furthermore, the distribution of endobionts such as mycoplasmas or Trichomonas vaginalis virus (TVV) in the T. vaginalis metapopulation is poorly characterised. RESULTS: We assayed the relationship between 20 strains of T. vaginalis from 8 countries using the Random Amplified Polymorphic DNA (RAPD) analysis with 27 random primers. The genealogical tree was constructed and its bootstrap values were computed using the program FreeTree. Using the permutation tail probability tests we found that the topology of the tree reflected both the pattern of resistance to metronidazole (the major anti trichomonal drug) (p < 0.01) and the pattern of infection of strains by mycoplasmas (p < 0.05). However, the tree did not reflect pattern of virulence, geographic origin or infection by TVV. Despite low bootstrap support for many branches, the significant clustering of strains with similar drug susceptibility suggests that the tree approaches the true genealogy of strains. The clustering of mycoplasma positive strains may be an experimental artifact, caused by shared RAPD characters which are dependent on the presence of mycoplasma DNA. CONCLUSIONS: Our results confirmed both the suitability of the RAPD technique for genealogical studies in T. vaginalis and previous conclusions on the relatedness of metronidazol resistant strains. However, our studies indicate that testing analysed strains for the presence of endobionts and assessment of the robustness of tree topologies by bootstrap analysis seem to be obligatory steps in such analyses. PMID- 11734060 TI - Genome trees constructed using five different approaches suggest new major bacterial clades. AB - BACKGROUND: The availability of multiple complete genome sequences from diverse taxa prompts the development of new phylogenetic approaches, which attempt to incorporate information derived from comparative analysis of complete gene sets or large subsets thereof. Such attempts are particularly relevant because of the major role of horizontal gene transfer and lineage-specific gene loss, at least in the evolution of prokaryotes. RESULTS: Five largely independent approaches were employed to construct trees for completely sequenced bacterial and archaeal genomes: i) presence-absence of genomes in clusters of orthologous genes; ii) conservation of local gene order (gene pairs) among prokaryotic genomes; iii) parameters of identity distribution for probable orthologs; iv) analysis of concatenated alignments of ribosomal proteins; v) comparison of trees constructed for multiple protein families. All constructed trees support the separation of the two primary prokaryotic domains, bacteria and archaea, as well as some terminal bifurcations within the bacterial and archaeal domains. Beyond these obvious groupings, the trees made with different methods appeared to differ substantially in terms of the relative contributions of phylogenetic relationships and similarities in gene repertoires caused by similar life styles and horizontal gene transfer to the tree topology. The trees based on presence absence of genomes in orthologous clusters and the trees based on conserved gene pairs appear to be strongly affected by gene loss and horizontal gene transfer. The trees based on identity distributions for orthologs and particularly the tree made of concatenated ribosomal protein sequences seemed to carry a stronger phylogenetic signal. The latter tree supported three potential high-level bacterial clades,: i) Chlamydia-Spirochetes, ii) Thermotogales-Aquificales (bacterial hyperthermophiles), and ii) Actinomycetes-Deinococcales-Cyanobacteria. The latter group also appeared to join the low-GC Gram-positive bacteria at a deeper tree node. These new groupings of bacteria were supported by the analysis of alternative topologies in the concatenated ribosomal protein tree using the Kishino-Hasegawa test and by a census of the topologies of 132 individual groups of orthologous proteins. Additionally, the results of this analysis put into question the sister-group relationship between the two major archaeal groups, Euryarchaeota and Crenarchaeota, and suggest instead that Euryarchaeota might be a paraphyletic group with respect to Crenarchaeota. CONCLUSIONS: We conclude that, the extensive horizontal gene flow and lineage-specific gene loss notwithstanding, extension of phylogenetic analysis to the genome scale has the potential of uncovering deep evolutionary relationships between prokaryotic lineages. PMID- 11734061 TI - Discrimination of three mutational events that result in a disruption of the R122 primary autolysis site of the human cationic trypsinogen (PRSS1) by denaturing high performance liquid chromatography. AB - BACKGROUND: R122, the primary autolysis site of the human cationic trypsinogen (PRSS1), constitutes an important "self-destruct" or "fail-safe" defensive mechanism against premature trypsin activation within the pancreas. Disruption of this site by a missense mutation, R122H, was found to cause hereditary pancreatitis. In addition to a c.365G>A (CGC>CAC) single nucleotide substitution, a c.365 through 366GC>AT (CGC>CAT) gene conversion event in exon 3 of PRSS1 was also found to result in a R122H mutation. This imposes a serious concern on the genotyping of pancreatitis by a widely used polymerase chain reaction-restriction fragment length polymorphism assay, which could only detect the commonest c.365G>A variant. MATERIALS AND METHODS: DNA samples containing either the known c.365G>A or c.365 through 366GC>AT variant in exon 3 of PRSS1 were used as positive controls to establish a denaturing high performance liquid chromatography (DHPLC) assay. RESULTS: DHPLC could readily discriminate the two known different mutational events resulting in the R122H mutation. More importantly, under the same experimental conditions, it identified a further mutational event that also occurs in the R122 primary autolysis site but results in a different amino acid substitution: c.364C>T (CGC>TGC; R122C). CONCLUSIONS: A rapid, simple, and low-cost assay for detecting both the known and new mutations occuring in the R122 primary autolysis site of PRSS1 was established. In addition, the newly found R122C variant represents a likely pancreatitis predisposing mutation. PMID- 11734062 TI - Expression of c-myc is not critical for cell proliferation in established human leukemia lines. AB - BACKGROUND: A study was undertaken to resolve preliminary conflicting results on the proliferation of leukemia cells observed with different c-myc antisense oligonucleotides. RESULTS: RNase H-active, chimeric methylphosphonodiester / phosphodiester antisense oligodeoxynucleotides targeting bases 1147-1166 of c-myc mRNA downregulated c-Myc protein and induced apoptosis and cell cycle arrest respectively in cultures of MOLT-4 and KYO1 human leukemia cells. In contrast, an RNase H-inactive, morpholino antisense oligonucleotide analogue 28-mer, simultaneously targeting the exon 2 splice acceptor site and initiation codon, reduced c-Myc protein to barely detectable levels but did not affect cell proliferation in these or other leukemia lines. The RNase H-active oligodeoxynucleotide 20-mers contained the phosphodiester linked motif CGTTG, which as an apoptosis inducing CpG oligodeoxynucleotide 5-mer of sequence type CGNNN (N = A, G, C, or T) had potent activity against MOLT-4 cells. The 5-mer mimicked the antiproliferative effects of the 20-mer in the absence of any antisense activity against c-myc mRNA, while the latter still reduced expression of c-myc in a subline of MOLT-4 cells that had been selected for resistance to CGTTA, but in this case the oligodeoxynucleotide failed to induce apoptosis or cell cycle arrest. CONCLUSIONS: We conclude that the biological activity of the chimeric c-myc antisense 20-mers resulted from a non-antisense mechanism related to the CGTTG motif contained within the sequence, and not through downregulation of c-myc. Although the oncogene may have been implicated in the etiology of the original leukemias, expression of c-myc is apparently no longer required to sustain continuous cell proliferation in these culture lines. PMID- 11734063 TI - Generation of choline for acetylcholine synthesis by phospholipase D isoforms. AB - BACKGROUND: In cholinergic neurons, the hydrolysis of phosphatidylcholine (PC) by a phospholipase D (PLD)-type enzyme generates some of the precursor choline used for the synthesis of the neurotransmitter acetylcholine (ACh). We sought to determine the molecular identity of the relevant PLD using murine basal forebrain cholinergic SN56 cells in which the expression and activity of the two PLD isoforms, PLD1 and PLD2, were experimentally modified. ACh levels were examined in cells incubated in a choline-free medium, to ensure that their ACh was synthesized entirely from intracellular choline. RESULTS: PLD2, but not PLD1, mRNA and protein were detected in these cells and endogenous PLD activity and ACh synthesis were stimulated by phorbol 12-myristate 13-acetate (PMA). Introduction of a PLD2 antisense oligonucleotide into the cells reduced PLD2 mRNA and protein expression by approximately 30%. The PLD2 antisense oligomer similarly reduced basal- and PMA-stimulated PLD activity and ACh levels. Overexpression of mouse PLD2 by transient transfection increased basal- (by 74%) and PMA-stimulated (by 3.2-fold) PLD activity. Moreover, PLD2 transfection increased ACh levels by 26% in the absence of PMA and by 2.1-fold in the presence of PMA. Overexpression of human PLD1 by transient transfection increased PLD activity by 4.6-fold and ACh synthesis by 2.3-fold in the presence of PMA as compared to controls. CONCLUSIONS: These data identify PLD2 as the endogenous enzyme that hydrolyzes PC to generate choline for ACh synthesis in cholinergic cells, and indicate that in a model system choline generated by PLD1 may also be used for this purpose. PMID- 11734065 TI - 5-fluorouracil-induced cardiotoxicity mimicking myocardial infarction: a case report. AB - BACKGROUND: Severe cardiotoxicity is a documented, but very unusual side-effect of intravenous 5-fluorouracil therapy. The mechanism producing cardiotoxicity is poorly understood. CASE PRESENTATION: A case of 5-fluorouracil-induced cardiotoxicity, possibly due to coronary artery spasm, and mimicking acute anterolateral myocardial infarction is presented and discussed. Electrocardiographs highlighting the severity of the presentation are included in the report along with coronary angiograms demonstrating the absence of significant coronary atherosclerosis. CONCLUSION: Severe 5-fluorouracil-induced cardiotoxicity is rare, but can be severe and may mimic acute myocardial infarction, leading to diagnostic and therapeutic dilemmas. Readministration of 5 fluorouracil is not advised following an episode of cardiotoxicity. PMID- 11734064 TI - Treatment of established postoperative nausea and vomiting: a quantitative systematic review. AB - BACKGROUND: The relative efficacy of antiemetics for the treatment of postoperative nausea and vomiting (PONV) is poorly understood. METHODS: Systematic search (MEDLINE, Embase, Cochrane Library, bibliographies, any language, to 8.2000) for randomised comparisons of antiemetics with any comparator for the treatment of established PONV. Dichotomous data on prevention of further nausea and vomiting, and on side effects were combined using a fixed effect model. RESULTS: In seven trials (1,267 patients), 11 different antiemetics were tested without placebos; these data were not further analysed. Eighteen trials (3,809) had placebo controls. Dolasetron 12.5-100 mg, granisetron 0.1-3 mg, tropisetron 0.5-5 mg, and ondansetron 1-8 mg prevented further vomiting with little evidence of dose-responsiveness; with all regimens, absolute risk reductions compared with placebo were 20%-30%. The anti-nausea effect was less pronounced. Headache was dose-dependent. Results on propofol were contradictory. The NK1 antagonist GR205171, isopropyl alcohol vapor, metoclopramide, domperidone, and midazolam were tested in one trial each with a limited number of patients. CONCLUSIONS: Of 100 vomiting surgical patients receiving a 5-HT3 receptor antagonist, 20 to 30 will stop vomiting who would not have done so had they received a placebo; less will profit from the anti-nausea effect. There is a lack of evidence for a clinically relevant dose-response; minimal effective doses may be used. There is a discrepancy between the plethora of trials on prevention of PONV and the paucity of trials on treatment of established symptoms. Valid data on the therapeutic efficacy of classic antiemetics, which have been used for decades, are needed. PMID- 11734066 TI - Updating a systematic review--what difference did it make? Case study of nicotine replacement therapy. AB - AIMS: To examine the effect of updating a systematic review of nicotine replacement therapy on its contents and conclusions. METHODS: We examined the effects of regular updating of a systematic review of nicotine replacement therapy for smoking cessation. We considered two outcomes. First, we assessed the effect of adding new data to meta-analyses, comparing results in 2000 with the results in 1994. Second, we assessed qualitatively the ways in which the nature of the questions addressed by the review had changed between the two dates. For the first outcome, we compared the number of trials, the pooled estimate of effect using the odds ratio, and the results of pre-specified subgroup analyses, for nicotine gum and patch separately. Using a test for interaction, we assessed whether differences between estimates were statistically significant. RESULTS: There were ten new trials of nicotine gum between 1994 and 2000, and the meta analytic effect changed little. For the nicotine patch the number of trials increased from 9 to 30, and the meta-analytic effect fell from 2.07 (95% CI 1.64 2.62) to 1.73 (95% CI 1.56 - 1.93). Apparent differences in relative effect in sub-groups found in 1994 were not found in 2000. The updated systematic review addressed a number of questions not identified in the original version. CONCLUSIONS: Updating the meta-analyses lead to a more precise estimate of the likely effect of the nicotine patch, but the clinical message was unchanged. Further placebo controlled NRT trials are not likely to add to the evidence base. It is questionable whether updating the meta-analyses to include them is worthwhile. The content of the systematic review has, however, changed, with the addition of data addressing questions not considered in the original review. There is a tension between the principle of identifying the important questions prior to conducting a review, and keeping the review up to date as primary research identifies new avenues of enquiry. PMID- 11734067 TI - Early bronchopulmonary involvement in Crohn disease: a case report. AB - BACKGROUND: Bronchopulmonary manifestations of Crohn disease have been rarely described in children, including both subclinical pulmonary involvement and severe lung disease. CASE PRESENTATION: A 6.5-year-old girl is described with early recurrent bronchopulmonary symptoms both at presentation and in the quiescent phase of Crohn disease. Pulmonary function tests (lung volumes and flows, bronchial reactivity and carbon monoxide diffusing capacity) were normal. Bronchoalveolar cytology showed increased (30%) lymphocyte counts and bronchial biopsy revealed thickening of basal membrane and active chronic inflammation. CONCLUSIONS: Clinical and histological findings in our young patient suggest involvement of both distal and central airways in an early phase of lung disease. The pathogenesis of Crohn disease-associated lung disorders is discussed with reference to the available literature. A low threshold for pulmonary evaluation seems to be advisable in all children with CD. PMID- 11734068 TI - Observer variation in chest radiography of acute lower respiratory infections in children: a systematic review. AB - BACKGROUND: Knowledge of the accuracy of chest radiograph findings in acute lower respiratory infection in children is important when making clinical decisions. METHODS: I conducted a systematic review of agreement between and within observers in the detection of radiographic features of acute lower respiratory infections in children, and described the quality of the design and reporting of studies, whether included or excluded from the review.Included studies were those of observer variation in the interpretation of radiographic features of lower respiratory infection in children (neonatal nurseries excluded) in which radiographs were read independently and a clinical population was studied. I searched MEDLINE, HealthSTAR and HSRPROJ databases (1966 to 1999), handsearched the reference lists of identified papers and contacted authors of identified studies. I performed the data extraction alone. RESULTS: Ten studies of observer interpretation of radiographic features of lower respiratory infection in children were identified. Seven of the studies satisfied four or more of the seven design and reporting criteria. Six studies met the inclusion criteria for the review. Inter-observer agreement varied with the radiographic feature examined. Kappa statistics ranged from around 0.80 for individual radiographic features to 0.27-0.38 for bacterial vs viral etiology. CONCLUSIONS: Little information was identified on observer agreement on radiographic features of lower respiratory tract infections in children. Agreement varied with the features assessed from "fair" to "very good". Aspects of the quality of the methods and reporting need attention in future studies, particularly the description of criteria for radiographic features. PMID- 11734069 TI - Re-HEDP : pharmacokinetic characterization, clinical and dosimetric evaluation in osseous metastatic patients with two levels of radiopharmaceutical dose. AB - BACKGROUND: A study for pain relief therapy with 188Re-HEDP was done in patients with bone metastases secondary to breast and prostate cancer. MATERIALS AND METHODS: Patients received 1.3 or 2.2 GBq, in single or multiple doses. Platelets, white and red cells were evaluated during 11 weeks. Pharmacokinetic characterization was done from blood and urine samples for 5 patients along 24 hours. Urinary excretion was evaluated in other 16 patients during 6 hours. Bone uptake was estimated as remaining activity in whole body. Scintigraphic images were acquired at 2 and 24 hs post-administration. Absorbed dose in bone marrow was estimated with Mirdose3. Analgesics intake and pain score were daily recorded. Tumour markers (PSA, and Tn-structure) were monitored in 9 patients during 4 to 6 months. Single doses of low activity (1.3 GBq) were given to twelve patients. Nine patients received multiple doses. RESULTS: All except one patient had normal levels of platelets, white and red cells. Remaining dose in blood at 2 hours was 9%. Urinary elimination was 58%. Bone uptake at 24 hours was 43% (mean value; n = 5). No changes of the haematological parameters were detected along follow-up period. Pain relief was evidenced by decrease or supression of opioid analgesic and by subjective index. PSA showed a decrease in prostate cancer patients (n = 4). Tn-structure showed a significant increase after 4 to 8 months. CONCLUSION: Single or multiple dose scheme could be safely used, with administered activity of 188Re-HEDP up to 60 mCi, with low bone marrow absorbed doses. PMID- 11734070 TI - Macrofilaricides and onchocerciasis control, mathematical modelling of the prospects for elimination. AB - BACKGROUND: In most endemic parts of the world, onchocerciasis (river blindness) control relies, or will soon rely, exclusively on mass treatment with the microfilaricide ivermectin. Worldwide eradication of the parasite by means of this drug is unlikely. Macrofilaricidal drugs are currently being developed for human use. METHODS: We used ONCHOSIM, a microsimulation mathematical model of the dynamics of onchocerciasis transmission, to explore the potentials of a hypothetical macrofilaricidal drug for the elimination of onchocerciasis under different epidemiological conditions, as characterized by previous intervention strategies, vectorial capacity and levels of coverage. RESULTS: With a high vector biting rate and poor coverage, a very effective macrofilaricide would appear to have a substantially higher potential for achieving elimination of the parasite than does ivermectin. CONCLUSIONS: Macrofilaricides have a substantially higher potential for achieving onchocerciasis elimination than ivermectin, but high coverage levels are still key. When these drugs become available, onchocerciasis elimination strategies should be reconsidered. In view of the impact of control efforts preceding the introduction of macrofilaricides on the success of elimination, it is important to sustain current control efforts. PMID- 11734071 TI - Knowledge and perception about stroke among an Australian urban population. AB - BACKGROUND: The aim of the study was to measure knowledge about the symptoms, prevalence and natural history of stroke; the level of concern about having a stroke; understanding of the possibilities for preventing stroke, and the relationship between age, sex, country of origin, educational level, income, self reported risk factors, and the above factors. METHODS: A random sample of households was selected from an electronic telephone directory in Newcastle and Lake Macquarie area of New South Wales, Australia, between 10 September and 13 October 1999. Within each household the person who was between 18 and 80 years of age and who had the next birthday was eligible to participate in the study (1325 households were eligible). The response rate was 62%. RESULTS: The most common symptoms of stroke listed by respondents were "Sudden difficulty of speaking, understanding or reading" identified by 60.1% of the respondents, and "paralysis on one side of body" identified by 42.0% of the respondents. The level of knowledge of the prevalence of a stroke, full recovery after the stroke, and death from stroke was low and generally overestimated. 69.9% of the respondents considered strokes as being either moderately or totally preventable. There were few predictors of knowledge. CONCLUSION: The study suggests that educational strategies may be required to improve knowledge about a wide range of issues concerning stroke in the community, as a prelude to developing preventive programmes. PMID- 11734072 TI - Protocol of a prospective cohort study of the effect of different methods of drainage of neuropathic bladder on occurrence of symptomatic urinary infection, and adverse events related to the urinary drainage system in spinal cord injury patients. AB - BACKGROUND: To present a protocol of a prospective, cohort study in which four groups of spinal cord injury (SCI) patients will participate. (Patients with indwelling urethral catheter; patients who perform intermittent catheterisation without wearing a penile sheath; patients who perform intermittent catheterisation and wear penile sheath as well; and patients with penile sheath drainage). OBJECTIVES: (1) What is the incidence of symptomatic urinary infection in men with spinal cord injury who use different types of bladder drainage? (2) Which are predisposing factors for the occurrence of symptomatic urinary infection in men with spinal cord injury who practise different methods of bladder drainage? (3) What is the incidence of catheter and urinary drainage system-related adverse events in the four groups of SCI patients? PATIENTS: The criteria for inclusion are as follow: (1) Male patients with neuropathic bladder due to spinal cord injury, who are registered with the Regional Spinal Injuries Centre, Southport, England. (2) Age: 18 years or above. (3) Patients who are willing to give informed consent for participation in the study. (4) Patients willing to be contacted every two weeks by a staff of the spinal unit for 36 months. (5) Patients who are willing to maintain an accurate record of adverse events related to urinary catheter and urinary drainage system and predisposing factors for the occurrence of symptomatic urinary infection. (6) Patients, who are stabilised in a particular method of bladder drainage, and therefore, unlikely to make a permanent change in the method of bladder drainage (e.g. from penile sheath drainage to the use of long-term indwelling catheter) during a foreseeable future. METHODS: The participants will be observed for a period of 36 months. A staff of the spinal injuries unit will contact the participants by telephone every two weeks on a mutually agreed day and time. The information obtained during this standardised telephonic interview conducted once in two weeks will be entered in a database. When a participant develops symptom(s) suggestive of urinary infection, he will undergo urine and blood tests, and imaging studies of the urinary tract. CONCLUSION: This study will provide information regarding the occurrence of symptomatic urinary infection, predisposing factors for development of urinary infection, and adverse events related to urinary catheter and urinary drainage system in SCI patients using different methods of bladder drainage. PMID- 11734073 TI - Effect of alpha-tocopherol on pulmonary antioxidant defence system and lipid peroxidation in cigarette smoke inhaling mice. AB - BACKGROUND: Free radicals generated in biological systems by cigarette smoke (CS) inhalation can cause oxidative stress in tissues, resulting in lipid peroxidation (LPO). In view of the antioxidant properties of alpha-tocopherol (AT), in the present study, effects of AT on antioxidant defence system and LPO were investigated in mice inhaling CS for different time intervals. RESULTS: Male Balb/c mice were fed orally with AT (5 I.U./Kg.b.wt.) and /or exposed to CS for 2, 4, 6 or 8 weeks. No effect was observed on body growth, diet consumption, water intake and lung weight due to AT and /or CS treatment in any of the groups as compared to their control counterparts. After two weeks of treatment, no change in LPO, reduced glutathione (GSH) levels and antioxidant enzymes were observed except for glutathione reductase (GR) which increased in all the treated groups. A significant increase in pulmonary LPO levels was observed in mice exposed to CS inhalation for 4, 6 or 8 weeks. There was a gradual increase in the LPO levels as the extent of CS inhalation increased from 4 to 8 weeks. However, the extent of increase in LPO levels due to CS exposure for 4, 6 or 8 weeks in the mice treated with AT was comparatively less. A significant decrease in the GSH levels was also observed in all the animals exposed to CS for 4, 6 or 8 weeks. There was a significant increase in the activities of catalase, glutathione peroxidase (GSH-Px) and GR observed in all the groups exposed to CS for 4,6 or 8 weeks. The increase in above antioxidant enzymes seems to be insufficient to combat the oxidative stress posed by CS inhalation. There was a marked decrease observed in the LPO levels in the animals treated with AT alone for 4, 6, or 8 weeks, when compared to their control counterparts. However, the supplementation of AT for 4, 6 or 8 weeks demonstrated a significant increase in GSH levels. CONCLUSION: It appears from our studies that AT exhibits its antioxidant role either directly by scavenging the oxidative species or indirectly by modulating the GSH levels. PMID- 11734074 TI - The temperature-dependent modulation of an inhibitory circuit in hippocampal slices as revealed by a population spike recording is mediated by extracellular adenosine. AB - We examined the effects of temperature on excitatory synaptic transmission and the recurrent inhibitory loop in CA1 neurons in guinea pig hippocampal slices. Increasing the temperature of the perfusing medium from 30 to 49 degrees C resulted in attenuation of both the amplitude of the synaptically evoked CA1 population spikes and the paired-pulse inhibition (PPI) of the spikes. A bath application of 2 microM picrotoxin, a gamma-aminobutyric acid receptor antagonist, did not affect the amplitude of the CA1 population spikes, but it significantly reduced PPI during the early heating phase (30-32 degrees C). In contrast, the application of 1 mM theophylline or 50 microM 8-phenyltheophylline, a selective adenosine A1 receptor antagonist, resulted in significant augmentation of the PPI during the early phase of hyperthermia (30-34 degrees C) and a significant increase in the amplitude of the CA1 population spikes at higher temperatures (34-43 degrees C). These results suggest that increased activation of adenosine A1 receptors in response to a temperature increase depresses not only excitatory synaptic responses, but also the strength of the inhibitory circuit in CA1 neurons. Furthermore, hyperexcitability of CA1 pyramidal neurons was seen in the middle of the heating range (34-38 degrees C), excitatory responses still being present, but the strength of the inhibitory circuit significantly reduced. PMID- 11734075 TI - Effects of pCO(2) on the CSF turnover rate in T(1)-weighted magnetic resonance imaging. AB - The cerebrospinal fluid (CSF) secretion of rat was monitored by longitudinal relaxation time-weighted magnetic resonance imaging (T(1)-weighted MRI) in combination with a ventricular injection of a T(1)-relaxation reagent: gadolinium diethylene triamine-N,N,N',N",N"-pentaacetic acid (Gd-DTPA). A cannula was inserted in the left lateral ventricle, and 5 microl of 8.5 mM Gd-DTPA was injected as a CSF marker. Changes in the image intensity of the CSF were measured every 30 s, and the turnover rate of CSF (k) in the left lateral ventricle was obtained from the dilution of Gd-DTPA, based on the assumption of a single compartment model. In the control conditions, k was 0.158 +/- 0.009 min(-1) at an arterial blood CO(2) tension (pCO(2)) of 38.6 +/- 2.2 mmHg (n = 10), which corresponds to the CSF secretion rate of 3.6 microl min(-1). The k value was decreased (0.078 +/- 0.010 min(-1), n = 4) by a carbonic-anhydrase inhibitor (acetazolamide). The turnover rate was decreased by hypocapnia (0.094 +/- 0.019 min(-1), pCO(2) = 24.7 +/- 2.9 mmHg, n = 4), and it increased gradually and reached a plateau level as a result of hypercapnia (0.194 +/- 0.011 min(-1), pCO(2) = 104.5 +/- 7.1 mmHg, n = 10). These results suggested that CO(2) upregulates the secretion of CSF in the rat. PMID- 11734076 TI - Relationship between mitochondrial DNA polymorphism and the individual differences in aerobic performance. AB - This study focused on the mitochondrial DNA (mtDNA) as the genetic factor most likely to bring about the individual difference in endurance capacity or its trainability. Platelets contain mtDNA but no nuclear DNA, whereas rho(0)-HeLa cells have nuclear DNA but no mtDNA. The oxidative capacity of mitochondria in the cultured cells, which were fused rho(0)-HeLa cell with platelets obtained from individual subjects (the so-called "cybrids"), reflects the individual mtDNA polymorphism in the gene-coding region. The purpose of this study was to investigate the relationship between the oxidative capacity of cybrids and the individual difference in endurance capacity, or its trainability. Forty-one sedentary young males took part in an 8-week endurance training program. They were determined by using their VO(2 max) as an index of endurance capacity on an ergocycle before and after the endurance training program. The relations between VO(2 max) before endurance training or the change of it by endurance training and the oxidative capacity of cybrids were investigated. There was no relation between them, and two groups were drawn from all subjects, based on one standard division of their initial VO(2 max): the higher pre-VO(2 max) group (n = 6) and the lower pre-VO(2 max) group (n = 5) (51.8 +/- 3.5 ml/min/kg vs. 33.3 +/- 3.8 ml/min/kg, p < 0.01). No significant difference was found between the O(2) consumption of the cybrids in the higher initial VO(2 max) group and that in the lower initial VO(2 max) group (16.3 +/- 4.9 vs. 15.9 +/- 2.0 nmol O(2)/min/10(7) cells, NS). Furthermore, neither the cytochrome c oxidase (COX) activity nor the complex I + III activity of cybrids showed a significant difference between the two groups. The oxidative capacity of cybrids between the high trainability group (n = 6) (Delta VO(2 max) 12.1 +/- 1.6 ml/min/kg) and the low trainability group (n = 9) (Delta VO(2 max) 2.3 +/- 0.5 ml/min/kg) was also similar. Thus the mtDNA polymorphism is very unlikely to relate to the individual difference in endurance capacity or its trainability in young sedentary healthy subjects. PMID- 11734077 TI - Ion channels permeable to monovalent and divalent cations: a single-file two-site channel model. AB - A cation channel from Tetrahymena cilia is permeable to both monovalent and divalent cations. A single-file two-site channel model was introduced for explaining the single channel currents of the channel in mixed solutions of K(+) and Ca(2+). In the model it was assumed that two potassium ions or one calcium ion can bind to the binding sites, and that the potassium ions between the binding sites are in a fast equilibrium condition. Single channel currents were calculated from the values of rate constants, ionic concentrations on both sides of the membrane, and the membrane voltages. This model could explain all the observed single channel currents of the channel in K(+) or Ca(2+) solution and in mixed solutions of K(+) and Ca(2+). The values of the reversal potential in the bi-ionic condition could distinguish this single-file two-site channel model from the single-site channel model or the model in which each ion permeates through the same channel independently (the Goldman-Hodgkin-Katz equation). Experimental data supported this model. PMID- 11734078 TI - Relationship between intracellular calcium and airway reactivity in guinea pigs. AB - The present study was carried out to examine the relationship between intracellular free calcium ion concentrations and its regulatory enzymes, sodium potassium adenosine triphosphatase (Na(+),K(+)-ATPase) and calcium adenosine triphosphatase (Ca(2+)-ATPase), with airway reactivity to inhaled histamine in guinea pigs. Forty-nine guinea pigs were included in this study. Of these, 34 animals responded to histamine bronchoprovocation challenge in vivo with a greater than 35% fall in specific airways conductance and were labeled as "reactive," and the remaining 15 were "nonreactive." The dose of histamine producing a 35% fall in specific airways conductance was labeled as ED(35) SGaw. The animals were then sacrificed, and the following biochemical measurements were carried out: intracellular free calcium ion concentrations [Ca(2+)](i) in leukocytes and isolated tracheal smooth muscle cells, activities of Na(+),K(+) ATPase and Ca(2+)-ATPase in tracheal homogenate, and plasma levels of lysophosphatidylcholine (LPC). Reactive guinea pigs showed significantly higher [Ca(2+)](i) and Na(+),K(+)-ATPase and Ca(2+)-ATPase activities. Airway reactivity (ED(35) SGaw) had significant negative correlation with [Ca(2+)](i), with activities of each of the ATPases and with plasma lysophosphatidylcholine. It is concluded that the level of [Ca(2+)](i) is an important determinant of airway reactivity. Intracellular calcium levels modulate airway response to histamine with higher levels being associated with greater reactivity. PMID- 11734079 TI - Importance of CCK-A receptor for gallbladder contraction and pancreatic secretion: a study in CCK-A receptor knockout mice. AB - Bile and pancreatic secretions were determined in a CCK-A receptor deficient mouse mutant generated by gene targeting in embryonic stem cells. The targeting vector contained lacZ and neo insertions in exon 2. Under the urethane anesthesia, the common bile duct was cannulated, and the mixture of bile pancreatic juice was collected every 30 min. After the 1 h basal secretion, CCK-8 (0.5 and 1.0 nmol/kg), acetylcholine (500 nmol/kg), and neuromedin C (1.0 micromol/kg) were injected subcutaneously, and the secretions were collected following 1 h. Amylase and bile acid outputs were determined as parameters of pancreatic secretion and gallbladder contraction, respectively. In some CCK-A receptor (+/-) animals, LacZ staining was performed. CCK-8 significantly increased amylase and bile acid outputs in CCK-A receptor (+/+) and (+/-) mice, whereas no response was observed in (-/-) mice. Neuromedin C and acetylcholine increased amylase secretion in CCK-A receptor (-/-) mice similar to (+/-) and (+/+) mice. The same doses of neuromedin C and acetylcholine could not increase bile acid secretion. The gallbladder smooth muscles, pancreatic acinar cells, duct cells, and islets were stained by LacZ. CCK and CCK-A receptor are important for pancreatic secretion and gallbladder contraction. Neuromedin C and acetylcholine may compensate pancreatic function, but not gallbladder contraction. PMID- 11734080 TI - Reflex responses in plasma catecholamines caused by static contraction of skeletal muscle. AB - To examine a hypothesis of whether static muscle contraction produces a release of catecholamines from the adrenal medulla via reflex stimulation of preganglionic adrenal sympathetic nerve activity induced by receptors in the contracting muscle, we compared the reflex responses in a concentration of epinephrine (Ep) and norepinephrine (NEp) in arterial plasma during static contraction and during a mechanical stretch of the hindlimb triceps surae muscle in anesthetized cats. Static contraction was evoked by electrically stimulating the peripheral ends of the cut L(7) and S(1) ventral roots at 20 or 40 Hz. Mean arterial pressure (MAP) and heart rate (HR) increased 23 +/- 3.1 mmHg and 19 +/- 4.3 beats/min during static contraction. Ep in arterial plasma increased 0.18 +/- 0.072 ng/ml over the control of 0.14 +/- 0.051 ng/ml within 1 min from the onset of static contraction, and NEp increased 0.47 +/- 0.087 ng/ml over the control of 0.71 +/- 0.108 ng/ml. Following a neuromuscular blockade, although the same ventral root stimulation failed to produce the cardiovascular and plasma catecholamine responses, the mechanical stretch of the muscle increased MAP, HR, and plasma Ep, but not plasma NEp. With bilateral adrenalectomy, the baseline Ep became negligible (0.012 +/- 0.001 ng/ml) and the baseline NEp was lowered to 0.52 +/- 0.109 ng/ml. Neither static contraction nor mechanical stretch produced significant responses in plasma Ep and NEp following the adrenalectomy. These results suggest that static muscle contraction augments preganglionic adrenal sympathetic nerve activity, which in turn secretes epinephrine from the adrenal medulla into plasma. A muscle mechanoreflex from the contracting muscle may play a role in stimulation of the adrenal sympathetic nerve activity. PMID- 11734081 TI - Regional difference of muscle oxygen saturation and blood volume during exercise determined by near infrared imaging device. AB - Using a near infrared (NIR) imaging device, we tested the hypothesis that regional differences in oxygen status could be detected in the gastrocnemius muscle during exercise and recovery. Six healthy subjects performed the standing plantar flexion exercises for 2 min; the frequency was one contraction per second. The NIR imaging device was placed over the medial head of the right gastrocnemius muscle and the signals from two optical sensors situated on the middle proximal and middle distal portions were used. The NIR-O(2) saturation (difference between deoxygenated and oxygenated Hb signals) and NIR-blood volume (sum of the oxygenated and deoxygenated Hb signals) were calculated in optical density units. Plantar flexion resulted in more deoxygenation during exercise and more reoxygenation during recovery in the distal portion compared with the proximal portion. The changes in NIR-O(2) between rest and a 2 min exercise, and between a 2 min exercise and a 3 min recovery were 0.11 and -0.23, respectively, in the distal portion, which were significantly larger than proximal values (0.05 and -0.10, p < 0.05). Plantar flexion resulted in lower NIR-blood volumes during exercise and greater recovery of blood after exercise in the distal portion compared with the proximal portion. The changes in NIR blood volume between rest and a 2 min exercise and between a 2 min exercise and a 3 min recovery were -0.19 and 0.31, respectively, in the distal portion, significantly larger than proximal values (-0.07 and 0.12, p < 0.05 for all comparisons). These findings indicate that the distal portion of the medial gastrocnemius had larger changes in NIR O(2) saturation and NIR-blood volume than the proximal portion had. This is consistent with the distal portion having a greater impairment of blood flow possibly because of the higher intramuscular pressure during exercise. IN CONCLUSION: (1) regional differences in oxygen status in the gastrocnemius muscle were detected with exercise, with the distal portion having greater NIR-O(2) saturation and NIR-blood volume changes, and (2) the NIR imaging device might be a useful method to detect the regional differences of oxygen status in the muscle. PMID- 11734082 TI - Role of blood volume in the age-associated decline in peak oxygen uptake in humans. AB - It has been reported that maximal oxygen uptake (VO(2 max)) is linearly correlated with blood volume (BV) in young people and that there is a reduction in VO(2 max) with aging. To examine the involvement of BV in the reduction of VO(2 max), we used an incremental cycle ergometer protocol in a semi-recumbent position to determine the relationship between peak oxygen uptake (VO(2 peak)) and BV in older subjects (69.1 +/- 1.0 years; n = 22), then compared that relationship with that in young subjects (22.3 +/- 0.5 years; n = 31). In the present study, VO(2 peak) and BV were significantly lower in the older subjects, compared with those in the young subjects. A linear correlation was demonstrated between the VO(2 peak) and BV in both the older (r = 0.705; p < 0.001) and the young (r = 0.681; p < 0.001) subjects within the groups. However, an analysis of covariance with BV as a covariate revealed that VO(2 peak) at a given BV was smaller in the older subjects than in the young subjects (p < 0.001), i.e., graphically, the regression line determined for the older subjects showed a downward shift. The decreased peak heart rate as a result of aging (153 +/- 3 beats/min in the older vs. 189 +/- 2 beats/min in the young subjects) contributed partly to this downward shift. These results suggest that the BV is an important determinant factor for VO(2 peak), especially within an age group, and that the age-associated decline of VO(2 peak) is also, to a relatively larger degree, because of factors other than BV and heart rate. PMID- 11734083 TI - Optical recording of spontaneous respiratory neuron activity in the rat brain stem. AB - We report on the optical imaging of spontaneous respiratory neuron bursts in the ventrolateral medulla (VLM) of medullary slices or brain stem-spinal cord preparations. A medullary slice with a thickness of 1.0-1.4 mm or brain stem spinal cord from 0- to 4-d-old rats was stained with fluorescent voltage sensitive dye, RH795. Optical signals were recorded as a fluorescence change by using an optical recording apparatus with a 128 x 128 photodiode array and a maximum time resolution of 0.6 ms. Motoneuronal activity was simultaneously recorded at the hypoglossal nerve roots or fourth cervical ventral roots. Fluorescence changes corresponding to the spontaneous inspiratory burst activity were detected in the hypoglossal nucleus and VLM in slice preparations, and in a limited area extending rostrocaudally in the VLM of the brain stem-spinal cord preparation. These measurements did not require signal averaging by multiple trials. Results suggest that inspiratory neurons are localized in more compact form at the level of the nucleus ambiguous than at the more rostral VLM, and that peak activity during the inspiratory phase propagates from the caudal to the rostral VLM. In 60% of brain stem-spinal cord preparations, weak and scattered fluorescence changes preceding the inspiratory burst activity were detected more predominantly in the rostral part of the VLM. The present findings show the feasibility of optical recordings for the in vitro analysis of spontaneous respiratory neuron activity in the medulla. PMID- 11734084 TI - Generator sources of EEG large waves elicited by mental stress of memory recall or mental calculation. AB - Preceding mental sweating response (MSR) during mental stress, small wavelets and large waves are observed on EEG. In the present study, locations of current sources of the large waves were estimated. The results revealed that the current dipole was estimated around the orbits, reflecting activity of the eyelid or eye muscles. PMID- 11734085 TI - Hypothalamic region facilitating shivering in rats. AB - Although the posterior part of the hypothalamus has long been considered important for thermoregulatory shivering, it is unknown whether the neurons there or the passing fibers are implicated in the response. Exposure of urethane anesthetized rats to cold (15-21 degrees C) elicits shivering. An injection of muscimol (0.5 mM), a GABA(A) receptor agonist, into the medial part of the hypothalamus, including the dorsomedial and posterior nuclei, suppressed the cold induced shivering. This result suggests that neurons having an excitatory effect on shivering are in this region of the hypothalamus. PMID- 11734086 TI - Pharmacogenomics and genetics of primary hypertension. PMID- 11734088 TI - Hypertension and dementia. AB - The increase in life expectancy is associated with a sharp rise in cognitive disorders, particularly after the age of 80 years. The identification and management of risk factors for these invalidating and distressing conditions must be considered a priority. The fact that antihypertensive treatment has been demonstrated to decrease that risk offers a new opportunity to reduce the prevalence of such related disorders and promote healthy aging. PMID- 11734089 TI - Quality-of-life measures as predictors of mortality and morbidity. AB - Poor quality of life assessed singly, using an instrument designed to assess quality of life, or variously, using assessments of job stress, depression, anxiety, hostility, anger, or life events, seems to be predictive of poor cardiovascular health and attendant morbidity and mortality. Since risk factors appear to cluster together, new studies may benefit from concentrating on assessing quality of life and health in multivariate domains, including both psychological and behavioral aspects of life, and validating instruments for use as predictive tools in the future. However, single questions on energy also appear useful as predictors. PMID- 11734090 TI - The relationship between quality of life and adherence to treatment. AB - Hypertension is a preventable condition, and the outcomes of clinical trials have established that its treatment reduces cardiovascular morbidity and mortality. Although awareness about the disease has improved in the past two decades, the reality is that many people remain untreated or not adequately controlled. The treatment of hypertension is usually long term, and its success will depend on the effects of the drug regimen on the patient's quality of life. The relationship between quality of life and compliance is complex and merits careful study. Monitoring quality of life may be one of the best ways of improving adherence to treatment. Therefore, when developing an approach to the treatment of hypertension, physicians should take into consideration the impact of different antihypertensives on the patient's overall well being, and along with the side effects and contraindications, quality-of-life issues may determine the choice of medication. PMID- 11734091 TI - Quality of life with nonpharmacologic treatment of hypertension. AB - Therapeutic treatment of hypertension has been achieved successfully with both pharmacologic and nonpharmacologic interventions. Clinical trials have shown that various approaches to treatment result in different levels of blood pressure reduction as well as varying effects on quality of life. Standardizing the approach to measuring quality of life would be beneficial to the assessment of treatment outcomes in hypertension trials. This article reviews some of the strengths and weaknesses of both pharmacologic and nonpharmacologic treatments of hypertension, with special emphasis placed on effects of quality of life. PMID- 11734092 TI - Some economic consequences of noncompliance. AB - The twin problems of poor compliance and poor persistence with prescribed antihypertensive drug regimens appear to be responsible for much of the huge shortfall in the proportion of hypertensives whose treatment brings their blood pressure down to satisfactory levels. A further problem is the confounding of nonresponse and poor compliance in patients with "drug-resistant hypertension," in that about half of such patients are poor compliers, whose response to simple regimens usually proves satisfactory once their compliance with prescribed regimens is corrected. Electronic means for compiling ambulatory patients' drug dosing histories have now made it both technically and economically feasible to distinguish clearly between noncompliers and nonresponders, which clinical judgment cannot do because it is no better at making this crucial distinction than a coin toss. With the advent of reliable, economical measurements of patient compliance with prescribed drug dosing regimens, we can probably eliminate most of the compliance problems. The problem awaiting us after that is poor persistence with prescribed regimens for antihypertensive and other cardiovascular medicines that are meant for long-term or life-long use. A recent study has shown that median persistence with fully reimbursed drugs of the statin class is only 6 months, which is about one fortieth of the length it should be to realize full benefits of such therapy. PMID- 11734093 TI - Actual practice in hypertension: implications for persistence with and effectiveness of therapy. AB - Hypertension is often poorly controlled despite its importance and despite the availability of very effective treatments. Many factors contribute to poor control, including failure of patients to attend physicians, failure of physicians to detect and adequately treat hypertension to goal levels, and failure of patients to persist with prescribed therapy. An under-recognized problem is the failure, both in writing and in the application by third party payers, of consensus guidelines to recognize the important difference between efficacy in clinical trials and effectiveness in clinical practice. The issue of applicability of guidelines to actual practice was studied in the context of a Canadian Family Medicine teaching practice. At the time of the study, consensus guidelines for management of hypertension recommended that drug therapy be initiated with a beta-blocker or diuretic, except for patients with complicated hypertension for whom these drug classes are relatively contraindicated. It was generally assumed that widespread use of other classes of drugs represented inappropriate and wasteful use of resources. By a retrospective chart audit, we determined the proportion of patients in a family practice for whom therapy with a beta-blocker or diuretic was contraindicated, and examined the compliance of the physicians in the practice with guidelines. We found that about half of hypertensives had conditions for which recommended therapy is not a beta-blocker or diuretic. Thus, failure of physicians to follow guidelines is apparently less inappropriate than is widely perceived. PMID- 11734094 TI - Adverse events, compliance, and changes in therapy. AB - Compliance with medical therapy represents a multifactorial problem with marked consequences for individual and public health. Among the many contributing factors, the choice of drug may also determine the degree of compliance. Various studies investigating either compliance or persistence with antihypertensive therapy using a variety of methods have suggested that adherence to therapy may show the following pattern: diuretics < beta-blockers < calcium channel blockers < ACE inhibitors. Furthermore, two recent studies have shown that therapy with angiotensin II receptor blockers may be associated with better long-term adherence than other antihypertensive monotherapies including angiotensin converting enzyme inhibitors. Since medication compliance with antihypertensive therapy, among other factors, closely correlates with the experience of side effects, it may be speculated that the different classes of antihypertensives may induce varying degrees of compliance and persistence due to their different tolerability profiles. Side effects may induce variable compliance and nonpersistence by yet another mechanism. Therapy turbulence, ie, any change in medication, is also associated with nonpersistence. Therefore, side effects may directly or indirectly (via inducing therapy turbulence) underly variable compliance and nonpersistence. PMID- 11734097 TI - What's new in pediatric hypertension? AB - Research activities in the field of pediatric hypertension have been increasing in recent years, leading to important new findings in the epidemiology, diagnosis, and treatment of hypertension in children. This review summarizes recent work in these areas, focusing on the epidemic of obesity-related hypertension in children, advances in the drug therapy of childhood hypertension, and the increasing use of ambulatory blood pressure monitoring in evaluation of hypertensive children. Each of these three areas will no doubt develop further over the years ahead, resulting in further advances in the field of childhood hypertension. PMID- 11734096 TI - Hypertension in hemodialysis patients. AB - Hypertension is very common and often poorly controlled in patients undergoing chronic hemodialysis. While high blood pressure has been documented to adversely impact several intermediate outcomes of cardiovascular disease, whether hypertension is an independent risk factor for mortality in this population is not clear. Expansion of extracellular fluid volume is the major pathophysiologic mechanism for the development of hypertension in these patients; however, alterations in other humoral mechanisms also play a significant role. Optimization of volume status is, therefore, the cornerstone of therapy with additional use of antihypertensive medications as needed. Good quality prospective studies are urgently needed to define the measurement techniques and blood pressure goals, and to develop therapeutic strategies for more effective management of hypertension in this high-risk population. PMID- 11734098 TI - Hypertension and the kidney. AB - Hypertension and kidney function are intimately related, with each having significant influences on the other. Given the major role played by the kidney in maintenance of extracellular fluid volume and peripheral vascular resistance, the kidney is justifiably a target of investigation to determine its potential role in essential hypertension. Conversely, hypertension is associated with progressive renal failure, and hypertension-associated end-stage renal disease is the second leading cause of end-stage renal disease in the United States. It is therefore important that we continue to investigate the hypertension/renal relationship in an effort to better understand the determinants of essential hypertension and to prevent a major cause of end-stage renal disease. PMID- 11734099 TI - Novel mechanisms contributing to cerebral vascular dysfunction during chronic hypertension. AB - Chronic hypertension is a major risk factor for numerous cardiovascular disorders and is strongly associated with stroke. Hypertension alters cerebral vascular structure and may have profound deleterious effects on cerebral vascular function, the underlying mechanisms of which are still not well understood. Recent findings have led to important developments in our understanding of novel areas of cerebral vascular biology. This review briefly examines new evidence for physiologic and pathologic roles of K(+) channels, the renin-angiotensin system and reactive oxygen species, and Rho and Rho-kinase in regulation of cerebral vascular tone. PMID- 11734100 TI - Progressive Myoclonic Epilepsies. AB - The treatment of progressive myoclonus epilepsy (PME) remains a major therapeutic challenge in neurology. Generalized convulsive seizures are often well controlled through classic antiepileptic drugs (AEDs) like valproate and clonazepam, whereas myoclonus, the main symptom that is affecting patients most in their daily life, is usually refractory to standard AEDs. Alternative therapy concepts have been and still are investigated. Among the new drugs, zonisamide and piracetam have shown the most promising results as add-on treatments. Other therapeutic approaches, like the use of antioxidants, 5-hydroxytryptophan (5-HTP), and baclofen should also be taken into consideration for the treatment of intractable cases of PME. Nonpharmacologic treatment options such as diet and physical therapy should always be considered, because they may save costs and side effects. In some instances, the occasional use of alcohol has shown beneficial effects. PMID- 11734101 TI - Epilepsy in Aging Populations. AB - Older adults pose special challenges when seizures or epilepsy occur. Increased risk for seizures is often unrecognized. Older adults may have milder epilepsy, and may not require or tolerate standard doses of anti-epileptic drugs (AEDs). Polypharmacy and drug interactions are potential problems, and altered protein binding, hepatic function, renal clearance, and gastric function can affect AED dosing. Side effects such as ataxia, somnolence, and confusion are of heightened concern in this group. Older adults typically suffer localization or focal onset seizures. Although many AEDs treat these types of seizures, the AED chosen for any given patient is determined by concomitant polypharmacy, side effect profile, as well as underlying medical conditions. Free and total AED levels may be necessary. The goal of seizure management for a hospitalized patient may be rapid seizure control. As the individual moves into the outpatient setting, AED therapy without side effects and with appropriate quality of life must be pursued. PMID- 11734102 TI - Epilepsy in Pregnant Women. AB - For women of childbearing potential with epilepsy, seizures should be controlled with the smallest dosage of anti-epileptic drug (AED). Treatment with monotherapy should be achieved, if possible. The possibility of AED withdrawal should be considered in appropriate clinical setting prior to conception, and the AED treatment should be optimized prior to conception. Many pregnancies are unplanned, underscoring the need for constant vigilance in streamlining the treatment regimen. Prenatal counseling becomes particularly important, in order that both the physician and patient have open communication and realistic expectations about the course and outcome of a potential pregnancy. All women of childbearing potential with epilepsy should be informed about the known rates of teratogenicity of AEDs, possibility of increased seizure frequency during pregnancy, and the risks of the pregnancy and labor. All of the conventional AEDs are associated with an increased risk of major and minor anomalies in the offspring and are categorized as US Food and Drug Administration class C or D. Polytherapy increases this risk. Valproic acid and carbamazepine are each associated with an increased risk of neural tube defects, and should be avoided by women with a family history of spina bifida. This combination should be avoided, if possible. When a woman with epilepsy presents with pregnancy, a monotherapy regimen should not be changed if the seizures are well controlled. Reducing the number of AEDs can be considered in case of polytherapy, if the seizures are well controlled. If seizures are poorly controlled, adequate seizure control is the primary goal. Serum AED levels should be documented prior to conception, and within each trimester. More frequent monitoring may be necessary in case of poorly controlled seizures. If seizures have occurred during pregnancy, therapeutic AED levels should be documented in the late third trimester, prior to delivery. Phenytoin levels should also include an unbound fraction ("free" level); other unbound AED levels are not generally available. The dose adjustment should be made taking the whole clinical picture into account. Vitamin K 10 mg per day orally should be administered in the last 4 weeks of pregnancy for women taking hepatic enzyme-inducing AEDS (phenytoin, phenobarbital, primidone, carbamazepine, topiramate, and oxcarbazepine). The newborn should receive vitamin K 1 mg intravenously or intramuscularly regardless of maternal AED exposure. PMID- 11734103 TI - Cognitive Impairment Following Traumatic Brain Injury. AB - Cognitive impairments due to traumatic brain injury (TBI) are substantial sources of morbidity for affected individuals, their family members, and society. Disturbances of attention, memory, and executive functioning are the most common neurocognitive consequences of TBI at all levels of severity. Disturbances of attention and memory are particularly problematic, as disruption of these relatively basic cognitive functions may cause or exacerbate additional disturbances in executive function, communication, and other relatively more complex cognitive functions. Because of the high rate of other physical, neurologic, and psychiatric syndromes following TBI, a thorough neuropsychiatric assessment of the patient is a prerequisite to the prescription of any treatment for impaired cognition. Psychostimulants and other dopaminergically active agents (eg, methylphenidate, dextroamphetamine, amantadine, levodopa/carbidopa, bromocriptine) may modestly improve arousal and speed of information processing, reduce distractibility, and improve some aspects of executive function. Cautious dosing (start-low and go-slow), frequent standardized assessment of effects and side effects, and monitoring for drug-drug interactions are recommended. Cognitive rehabilitation is useful for the treatment of memory impairments following TBI. Cognitive rehabilitation may also be useful for the treatment of impaired attention, interpersonal communication skills, and executive function following TBI. This form of treatment is most useful for patients with mild to moderate cognitive impairments, and may be particularly useful for those who are still relatively functionally independent and motivated to engage in and rehearse these strategies. Psychotherapy (eg, supportive, individual, cognitive behavioral, group, and family) is an important component of treatment. For patients with medication- and rehabilitation-refractory cognitive impairments, psychotherapy may be needed to assist both patients and families with adjustment to permanent disability. PMID- 11734104 TI - Emotional Disturbances Following Traumatic Brain Injury. AB - Mood disturbances are common sequelae of traumatic brain injury (TBI), but the scientific database for such disorders is very limited in descriptive, prognostic, and treatment data. Post-TBI symptoms often cross diagnostic boundaries and include cognitive loss, amotivation, psychosis, mood, changes, or other domains. The treating physician must be mindful that clear diagnostic boundaries may not exist. Premorbid level of functioning commonly affects post TBI level of functioning. When setting treatment goals, this must be considered. Patients who had lower levels of psychosocial functioning before the injury may not fare as well afterwards. Treatment of post-TBI mood symptoms should proceed after a full diagnostic work-up including imaging and electroencephalographic (EEG) studies, neuropsychologic testing, and physical and laboratory examinations. Once the diagnostic picture is established, treatment should then proceed with a multidisciplinary team (physician, social worker, neuropsychologist, and others). For the medications, consider both target symptoms and side effects; start medications with low doses and raise slowly, give full therapeutic trials before switching or adding second agents, avoid benzodiazepines if possible, limit anticholinergic or antidopaminergic agents, and avoid providing large quantities of lethal medications. When starting medications for the treatment of mood disorders following TBI, several general principles of treatment in this population should be considered, including: balancing treatment of target symptoms with the potential for adverse effects; making use of side effects to treat comorbid problems when present (ie, relatively antidepressant for depression and marked insomnia); using a "start low, go slow" approach; continuing dose escalation to full therapeutic levels (ie, completing therapeutic trials) before switching or adding augmenting agents; avoiding agents with predictable and functionally important adverse effects (ie, benzodiazepines, strongly anticholinergic or antidopaminergic agents); and avoiding prescription of large and potentially lethal quantities of medications. PMID- 11734105 TI - Sleep Disturbances Following Traumatic Brain Injury. AB - Sleep disorders commonly complicate the course following traumatic brain injury (TBI). Insomnia, excessive daytime somnolence and alteration of the sleep-wake schedule are common disturbances that affect the course of recovery and prognosis in TBI survivors. Few studies, however, have looked at the diagnosis and management of these disturbances in TBI. Early treatment of sleep disorders must be considered an integral part of the rehabilitation process. Recognition and management of comorbid medical or surgical diseases, assessment and treatment of associated psychiatric disorders, and awareness of other psychosocial stressors are mandatory steps in the management of sleep disturbances following TBI. In addition to pharmacologic therapy, nonpharmacologic approaches such as diet, environmental modification, and behavioral interventions are essential components in the management of sleep disturbances in TBI. Based on the evidence that sleep disturbances impact rehabilitation in TBI patients we support the need for ongoing studies in this area. PMID- 11734106 TI - Post-traumatic Headache. AB - Headache is the most common symptom after closed head injury, persisting for more than 2 months in 60% of patients. Rarely does headache occur in isolation. Cervical pain is a frequent accompaniment. Post-traumatic headache is often one of several symptoms of the postconcussive syndrome, and therefore may be accompanied by additional cognitive, behavioral, and somatic problems. Acute post traumatic headaches may begin at the time of injury and continue for up to 2 months post-injury. Although onset proximate to the time of injury is most common, any new headache type occurring within this period of time is referred to as an acute post-traumatic headache. If such headaches persist beyond the first two months post-injury, they are subsequently referred to as chronic post traumatic headaches. Over time, post-traumatic headaches may take on a pattern of daily occurrence. If aggressive treatment is initiated early, posttraumatic headache is less likely to become a permanent problem. Once "windup" of post traumatic headaches occurs, the cycle of ongoing headaches is more difficult to interrupt. The mechanism of post-traumatic headache is poorly understood. Trauma induced headaches are usually heterogeneous in nature, often including both tension-type pain and intermittent migraine-like attacks. Rebound-headaches may develop from overuse of analgesic medications, and the occurrence of such may complicate significantly the diagnosis of post-traumatic headache. Adequate treatment typically requires both "peripheral" and "central" measures. Understanding the general principles of treatment, especially appropriate use of preventive and abortive medications, will most usefully guide treatment. There is scant literature with which to direct treatment selection for post-traumatic headache. Consequently, treatments for post-traumatic headache are based on those prescribed for phenomenologically similar but etiologically distinct headache disorders. Delayed recovery from post-traumatic headache may be a result of inadequately aggressive or ineffective treatment, overuse of analgesic medications resulting in analgesia rebound phenomena, or comorbid psychiatric disorders (eg, post-traumatic stress disorder, insomnia, substance abuse, depression, or anxiety). PMID- 11734107 TI - Adjuvant therapy trials of radiotherapy and systemic treatment for breast cancer. PMID- 11734108 TI - Development of novel signal transduction inhibitors; recent phase I trials. PMID- 11734109 TI - Tyrosine kinase inhibitors: a clinical perspective. AB - Cancer treatment has so far been restricted to cytotoxic and hormonal agents. These have been of limited value in their efficacy and their toxicity profile. A new era of targeted therapies is rapidly evolving. A key target being actively pursued is the receptor tyrosine kinase. Several compounds that inhibit this target are in preclinical and clinical development. These compounds broadly fall into two categories: monoclonal antibodies and small-molecule inhibitors. The common targets are epidermal growth factor receptor, Bcr-Abl tyrosine kinase, vascular endothelial growth factor, fibroblast growth factor receptor, and platelet-derived growth factor. Two of these compounds, trastuzumab and imatinib mesylate, have been approved by the US Food and Drug Administration for use in specific indications. Other uses are being tested, such as imatinib for gastrointestinal stromal tumor. These compounds will alter cancer care as adjuncts to currently available treatment options. PMID- 11734110 TI - Inhibition of vascular endothelial growth factor in the treatment of solid tumors. AB - Vascular endothelial growth factor (VEGF) is the term used for a family of tumor derived angiogenic factors that mediate endothelial proliferation and vascular permeability. Preclinical models have demonstrated the essential nature of VEGF in the angiogenesis of solid tumor growth and metastasis, whereas pathologic investigations have revealed strong correlations between VEGF production, microvessel density, and overall aggressiveness of many human solid tumors. Recent advances in the understanding of the molecular mechanisms of VEGF action have led to successful models for intervention in VEGF-mediated pathways in therapy for solid tumors. These include antibodies to block the binding of VEGF to its cellular receptors, small-molecule chemical inhibitors of the tyrosine kinase functions of the VEGF receptors, and antisense nucleic acids to interfere with cellular production of VEGF. Clinical investigations are ongoing to test the value of VEGF-based intervention alone or in combination with other anticancer agents. PMID- 11734111 TI - Evolving therapies: farnesyltransferase inhibitors. AB - Farnesyltransferase inhibitors (FTIs) are compounds designed to interfere with the signal transduction of cancer cells containing ras gene mutations. Specifically, FTIs were designed to prevent the farnesylation of Ras and other intracellular proteins, and they have been shown to have an effect on malignant cell proliferation and survival. However, the actual intracellular target of FTIs and the cellular determinants of drug action that correlate with antitumor effects currently are unknown. The following are key questions relating to FTI cell biology and clinical development: 1) Are ras gene mutations required for FTIs to be effective? 2) Does the effect of FTI therapy depend on which ras isoform is active (ie, H-ras, K-ras4A/B, or N-ras) in cancer cells? 3) What level of farnesyltransferase inhibition is required for clinical effect? 4) What surrogate biomarkers can be used to evaluate the biologic effect of FTIs in ongoing clinical trials? 5) What is the mechanism of FTI antitumor activity? Although the preliminary results are encouraging, more understanding of the intracellular mechanism of the FTIs is needed to determine how best to administer them in patients with cancer and to test their antitumor effects. PMID- 11734112 TI - Protein kinase C inhibitors. AB - Protein kinase C (PKC) is a family of serine-threonine protein kinases that are involved in signal transduction pathways that regulate growth factor response, proliferation, and apoptosis. Its central role in these processes, which are closely involved in tumor initiation, progression, and response to antitumor agents, makes it an attractive therapeutic target in cancer. Despite initial activity seen in melanoma (bryostatin and UCN-01), non-Hodgkin's lymphoma (ISIS 3521, bryostatin, and UCN-01), and ovarian carcinoma (ISIS 3521 and bryostatin) in phase I studies, single-agent activity in those phase II studies reported to date has been limited. Preclinical data highlight a role for PKC in modulation of drug resistance and synergy with conventional cytotoxic drugs. A randomized phase III study of ISIS 3521 in combination with carboplatin and paclitaxel, compared with chemotherapy alone, in advanced non-small-cell lung cancer is underway. This paper reviews the rationale for using PKC inhibitors in cancer therapy, the challenges for clinical trial design, and the recent clinical experience with modulators of PKC activity. PMID- 11734113 TI - An update on epidermal growth factor receptor inhibitors. AB - The epidermal growth factor receptor (EGFR) is part of a family of plasma membrane receptor tyrosine kinases that control many important cellular functions, from cell growth and proliferation to cell death. Dysregulation of the EGFR signal transduction pathway has been implicated in tumorigenesis and cancer progression, making it a clinically relevant target for novel anticancer treatments. This paper reviews recent progress in the development of cancer therapies that are directed toward particular aspects of the extracellular and intracellular domains of EGFR. Promising new compounds in the advanced stages of clinical testing are emphasized. PMID- 11734115 TI - Prevention of radiation-induced oral mucositis with benzydamine. PMID- 11734114 TI - Thalidomide: an antineoplastic agent. AB - It has been more than three decades since the withdrawal of thalidomide from the marketplace. Thalidomide is attracting growing interest because of its reported immunomodulatory and anti-inflammatory properties. Current evidence indicates that thalidomide reduces the activity of the inflammatory cytokine tumor necrosis factor-alpha by accelerating the degradation of its messenger RNA. Thalidomide inhibits angiogenesis. Recently, thalidomide was approved for sale in the United States for the treatment of erythema nodosum leprosum, an inflammatory complication of Hansen's disease. Thalidomide has been used successfully in several other dermatologic disorders, including aphthous stomatitis, Behcet's syndrome, chronic cutaneous systemic lupus erythematosus, and graft-versus-host disease, the apparent shared characteristic of which is immune dysregulation. Many recent studies have evaluated thalidomide in patients with HIV infection, in which this drug is an efficacious agent against oral aphthous ulcers, HIV associated wasting syndrome, HIV-related diarrhea, and Kaposi's sarcoma. Only in the last several years has thalidomide been aggressively investigated for its antiangiogenic potential and immunomodulatory properties in various tumor types. Current research on thalidomide in oncology covers investigation in a wide range of both solid tumors and hematologic malignancies. PMID- 11734116 TI - Sarcomas of the head and neck. AB - Sarcomas of the head and neck are extremely rare, accounting for less than 1% of all neoplasms of the head and neck. These sarcomas arise in both soft tissues and bone and thus cannot be treated by a single approach. The clinical behavior of these tumors varies considerably. Patients with low-grade lesions are prone to local recurrence, whereas those with high-grade lesions develop both local recurrence and disseminated disease. An additional complicating factor is the multiple sites in which sarcomas may arise in the head and neck. All of these factors make it difficult to ascertain the optimal treatment approach for sarcomas. This article reviews the current literature (as well as the author's own experience) and provides a general treatment guideline for sarcomas of the head and neck. PMID- 11734117 TI - Cancer of the external auditory canal and temporal bone. AB - Malignant neoplasms involving the temporal bone are a relatively rare and often misdiagnosed disease. Staging of temporal bone cancer has proven difficult because of the small number of patients with this condition, the various histopathologic and histologic findings reported, and a lack of randomized trials. Of the various staging systems that have been proposed, the Pittsburgh classification appears to be the most widely accepted. A retrospective study of 31 patients with temporal bone malignancy at the University of Arkansas for Medical Sciences has led us to propose a modification of the Pittsburgh classification for early-stage lesions. This modification places more emphasis on the site of disease in the canal and less on the size of the primary tumor or degree of bony invasion. This review discusses this staging system, the management of these tumors in a multidisciplinary team approach, reconstructive options, and auditory rehabilitation. PMID- 11734118 TI - Management of the clinically negative (N0) neck. AB - The clinically negative (N0) neck is defined by its absence of palpable or radiographically suspicious lymph nodes. Management of patients staged N0 is controversial. Clinical studies have lacked sufficient power and follow-up time to show a survival benefit for any particular option. Current options include observation, elective neck irradiation, and elective neck dissection. Treatment plans are based on treatment of the primary tumor and surgeon preference. PMID- 11734120 TI - [Cell oxidative processes and antioxidant mucoactive drugs]. PMID- 11734119 TI - Photodynamic therapy in head and neck cancer. AB - Photodynamic therapy is a nonsurgical, minimally invasive technique that uses a light source to activate light-sensitive drugs or photosensitizers to treat cancer and other diseases. The effect on living organisms caused by light in the presence of a photosensitizer and oxygen is known as the photodynamic effect or photodynamic action. PMID- 11734121 TI - [Factor analysis of the health related quality of life of patients with stable chronic obstructive pulmonary disease]. AB - OBJECTIVE: To identify factors associated with the health-related quality of life (HRQL) perceived by patients with stable chronic obstructive pulmonary disease (COPD). PATIENTS AND METHODS: DESIGN: Prospective cohort study. SUBJECTS: Patients with stable COPD (n = 204, alpha = 0.05, d = 6.9%). MEASURES AND INTERVENTIONS: For every patient we recorded sociodemographic and treatment data,severity of dyspnea, and the results of gasometry and lung function and stress testing, as well as responses to the Chronic Respiratory Disease Questionnaire (CRDQ). After studying the associations between variables we carried out factor analysis of those that were significantly related to HRQL. RESULTS: Men made up 91.7% of the cohort. The mean age (standard deviation)was 71 (9), FEV1% was 45 (16) and FEV1/FVCwas 48 (10). HRQL was statically related to age, weight, body mass index (BMI), economic status, educational status, chronic treatment with oral corticosteroids, ODCA, PaO2, oxygen saturation, alveolar arterial oxygen gradient, absolute FEV1, FEV1%, post-bronchodilatation FEV1, absolute FVC, FVC%, distance walked in six minutes, oxygen saturation before the walking test and lowest oxygen saturation during the test,difference in dyspnea before and after the test measured on a visual analog scale. Factor analysis reduced the main variables to six, which explained 74.6% of the variance, as follows: Factor1 PaO2, oxygen saturation and alveolar-arterial oxygen gradient (33.7%); Factor 2 absolute FEV1,FEV1%, FEV1 after bronchodilatation, absolute FVC and FVC% (11.9%); Factor 3 eight and BMI (8.8%); Factor 4 age, severity of dyspnea and walking test distance (8.4%);Factor 5: corticosteroid dependence and domiciliary oxygen therapy(6.3%); and Factor 6: economic and educational status (5.5%). CONCLUSION: HRQL in stable COPD patients is partially related to severity of respiratory insufficiency, airflow obstruction, nutritional status, exercise capacity, severity of disease, and socioeconomic status. PMID- 11734123 TI - [Study of the personality of patients with spontaneous pneumothorax]. AB - INTRODUCTION: Medical psychology has contributed to a greater understanding of many diseases that are predominantly medical and has also helped to improve prognosis. This study explores a surgical entity, namely spontaneous pneumothorax. OBJECTIVES: The aim was to compare the personality, depression, anxiety and type-A behavior pattern in a group of 34 patients with spontaneous pneumothorax to a group of 33 control patients admitted for a variety of minor surgical procedures. MATERIAL AND METHODS: The following objective assessment instruments were used: Trait Anxiety Inventory, Beck Depression Inventory, Jenkins Activity Inventory, Eysenck Personality Questionnaire. The questionnaires were administered before the intervention of the surgeon and after an informative interview. RESULTS: The rate of type-A behavior was statistically different in the two groups. No differences were seen for personality, depression or anxiety. CONCLUSION: We conclude that type-A behavior patterns should be reduced in patients who suffer spontaneous pneumothorax in order to improve outcome. PMID- 11734122 TI - [Tuberculosis notification from 1987 to 1999 for the public health area of the community of Valencia (Spain)]. AB - OBJECTIVE: Under notification of tuberculosis (TB) is common worldwide but has hardly been studied in Spain. The objective of this study was to determine how many cases of TB are reported in our health care area. MATERIAL AND METHOD: Between 1987 and 1999 we carried out a direct study of the incidence and reporting of TB cases in our area (106,632 inhabitants on the census). Cases were taken from mandatory notification, pathology and bacteriology files and were then reviewed. RESULTS: Of the 410 TB cases diagnosed, 378 were residents of the area (annual incidence: 27/100,000 inhabitants), 223 of whom were reported (54%). Forty-eight patients were under 15 years of age. Sixty-seven percent of the pediatric cases and 53% of the adult cases were reported (n.s.). Of the 78 cases with HIV co-infection, only 32% were reported, in contrast with 60% of HIV negative cases (p < 0.001). Of the 300 cases with pulmonary involvement, 56% were reported, versus 50% of those with exclusively extrapulmonary involvement (n.s.). For the 149 with positive Ziehl-Neelsen stain, 61% were reported, in contrast with 51% of those who were smear negative (n.s.). Diagnosis was based on clinical signs in 65 cases (16%), 51% of which were reported, versus 55% of those in which diagnosis was based on bacteriology or histology consistent with TB (n.s.). Overall under notification tended to decrease during the study period (r = 0.73, p < 0.005). Under notification also tended to decrease for patients with pulmonary involvement (r = 0.83, p < 0.0001) and for those with positive Ziehl Neelsen staining of sputum (r = 0.79, p< 0.001). Likewise, under notification also decreased for HIV negative patients (r = 0.74, p < 0.02) but not significantly so for HIV positive patients r = 0.44, n.s.). CONCLUSIONS: Direct quantification of cases revealed a significant degree of under notification that is tending to decrease. Under notification is greatest for HIV-positive patients and is not changing. No differences were seen for smear positive and smear negative patients, by pulmonary or extra-pulmonary involvement, for clinical versus laboratory diagnosis, or for patients who were younger or older than 15 years of age. Reporting should be improved so that measures to decrease the incidence of TB can be taken. PMID- 11734124 TI - [Risk factors for bronchial asthma in patients with rhinitis]. AB - OBJECTIVES: To identify risk factors for bronchial asthma in a large sample of patients with rhinitis. PATIENTS AND METHODS: One thousand seven hundred sixty patients with rhinitis treated at the respiratory medicine out-patient service of Hospital Universitario Virgen de Rocio in Seville (Spain) in 1997 and 1998 were studied. Six hundred forty-one (36.4%) had isolated rhinitis and 1,119 (63.6%) had rhinitis and bronchial asthma. The following variables were analyzed for both groups: 1) age, 2) sex, 3) clinical diagnosis, 4) age of onset of symptoms, 5) a family history of asthma and/or atopy, 6) housing conditions, 7) smoking, 8) a history of skin allergy, 9) recurrent episodes of respiratory infection with wheezing during early childhood, 10) a diagnosis of nasosinus polyposis, 11) atopy and sensitivity (pollens and/or household allergens), 12) peripheral blood eosinophil count. RESULTS: The variables that best differentiated the group with rhinitis from the group with both rhinitis and asthma were age, family history of asthma and/or atopy, exposure to household humidity or damp, a history of skin allergy, recurrent episodes of respiratory infection with wheezing in early childhood, atopy, sensitivity to household allergens and peripheral blood eosinophil count. The probability of correctly classifying patients in the appropriate group using this model was 69.7%. Among atopic patients, the best predictive variables were the same, with the exception of household humidity/damp. The probability of correct classification using this model was 69.7%. CONCLUSIONS: Patients with rhinitis have risk factors for bronchial asthma. As many such patients as possible should be identified so that long-term follow-up can take place and strategies to prevent bronchial asthma can be implemented. PMID- 11734125 TI - [Current trends in the treatment of locally advanced non-small cell bronchopulmonary carcinoma]. PMID- 11734126 TI - [Transition from pediatric cystic fibrosis units to adult units]. PMID- 11734127 TI - [Chest surgery and radiology. Remembering Wilhelm Conrad Rontgen in the 106th anniversary of the discovery of x-rays]. PMID- 11734128 TI - [Bilateral diaphragm paralysis after cardiac surgery]. AB - We report two cardiac surgery patients on whom local hypothermia with slushed ice for myocardial protection was used. Bilateral diaphragm paralysis with respiratory failure occurred. In both cases, neurophysiologic studies have been used for the diagnosis. Prolonged ventilatory support was required for several weeks and they were totally recovered after months. PMID- 11734129 TI - [Pneumomediastinum after transbronchial needle aspiraton]. PMID- 11734130 TI - [COPD: reflections on cause and course]. PMID- 11734132 TI - [How long should respiratory physiotherapy last?]. PMID- 11734133 TI - [Continuing medical education. A task for scientific societies]. PMID- 11734134 TI - [Analysis of the impact of Archivos de Bronconeumologia through the Science Citation Index]. AB - OBJECTIVE: To describe the impact factor of Archivos de Bronconeumologia from 1997 until 2000 and to identify the patterns of citation of the journal and topics having the greatest impact. METHOD: SCISEARCH was used to locate citations of articles published by Archivos de Bronconeumologia between 1995 and 1999. The following data were collected for each article: year of publication, authors, journal, country of publication, language, specialty or specialties, institution(s), residence of the first author and topic. The impact factor was calculated as the ratio of citations received in one year by articles published in Archivos de Bronconeumologia during the two previous years and the total number of articles published by Archivos de Bronconeumologia over the two years under study. RESULTS: The impact factor of Archivos de Bronconeumologia was 0.107 in 1997, 0.089 in 1998, 0.105 in 1999 and 0.119 in 2000. Citations were found in a wide range of source journals, with respiratory system publications having little weight. Citations were made mainly by Spanish authors (75%) and self citation was restrained (21.1%). Topics related to tuberculosis and respiratory infections (23.6% of the citations received) and chronic obstructive pulmonary disease (12.5%) made the greatest impact. CONCLUSION: The impact factor of Archivos de Bronconeumologia is modest, although higher than those of some other publications included in Journal Citation Reports. PMID- 11734135 TI - [Prevalence of sleep apnea-hypopnea syndrome among long-haul professional drivers]. AB - Sleep apnea-hypopnea syndrome (SAHS) has been associated with traffic accidents. The aim of this study was to estimate the prevalence of SAHS and analyze risk factors. We studied 163 professional drivers (86.7%) of the 188 employed by 25 participating companies. The subjects completed a questionnaire on SAHS symptoms and risk factors and underwent physical examination and conventional nighttime polysomnographic testing. RESULTS: The prevalence of an apnea-hypopnea index (AHI) ( 5 was 25.2% (95% CI 18.7-32.5) among the drivers. The prevalence of SAHS was 8.6% (95% CI 3.4-12.1). The prevalence increased with age (p = 0.012). Sleepiness while driving or habitual snoring had a sensitivity of 67.5%, specificity of 62.6% and a positive predictive value of 38.6% for detecting SAHS. Logistic regression modelling showed that the risk factors were a body mass index over 29 kg/m2 (OR: 3.56, 95% CI 1.53-8.4) and sleepiness while driving (OR: 3.7, 95% CI: 1.303-10.3). CONCLUSION: These results suggest that detecting SAHS among drivers may be useful for preventing traffic accidents; a questionnaire on SAHS symptoms and objective measures such as polysomnography allow cases to be detected and treated. PMID- 11734136 TI - [Non-small cell bronchogenic carcinoma in advanced stages: prognostic value of weight loss and clinical implications]. AB - This study of advanced-stage non-small cell bronchogenic carcinoma aimed 1) to identify prognostic factors collected at the moment of diagnosis, 2) to determine whether weight loss is a useful parameter to screen for subjects who will receive greater benefit from anticancer therapy.Patients and methods. Eighty-one patients were enrolled after diagnosis of stage III-B and IV non-small cell bronchogenic carcinoma and levels of activity < 2 according to Eastern Co-operative Oncology Group classification. The variables studied were age, sex, smoking history, associated disease, clinical data (weight loss, dyspnea, superior vena cava syndrome), laboratory parameters [hemoglobin, serum albumin, total lymphocytes, serum lactate dehydrogenase (LDH), calcium and liver enzymes], tumor-node metastasis (TNM) staging, histologic type, activity, treatment received and survival in weeks. The results were analyzed in two groups: 1) the general group consisting of results for all patients, and 2) the no-weight-loss group consisting of results for those whose weight had been stable. Student t, chi squared, Kaplan Meier, log-rank and Cox's regression model were used to analyze data and survival. RESULTS: Mean survival was 29 weeks (21-37). Survival was significantly related to weight loss, total lymphocytes, serum LDH, TNM and activity level in the general group. Only two factors continued to have prognostic value in the multivariate study: weight loss (OR: 1.48 (1.14-1.92), p = 0.002) and TNM (OR: 0.72 (0.54-0.96), p = 0.02). Among the patients with no weight loss, treatment received and TNM were significantly related to survival in univariable analysis and in Cox's regression model. CONCLUSIONS: In our experience with advanced lung cancer subjected to anticancer therapy, the presence of weight loss is the variable with the greatest prognostic value, such that it may be useful to consider it routinely, along with activity level, to try to identify patients who will receive the most benefit from cytostatic treatment. PMID- 11734137 TI - [Pediatric lung resection. A case series and evaluation of postoperative lung function]. AB - The most common causes of pulmonary lobectomy in children are congenital lung malformations (CLM) and bronchiectasias. Our aim was to present the causes and clinical course and lung function of lobectomized patients. Between 1990 and July 1999 27 lobectomies were performed on patients whose ages ranged from newborn to 14 years. Lobectomies were performed to correct CLM in 124 cases and for acquired pulmonary disease (APD) in 13. Among CLM cases, half (n = 7) had cystic adenomatoid malformation. Among the APD patients, 10 had bronchiectasias, with etiological confirmation in 6 cases (3 secondary to serious adenovirus infection). Mean hospital stay was 4.6 days among those who experienced no postoperative complications. Symptoms resolved after surgery for most symptomatic patients. Lung function tests could be carried out with 8 patients over 6 years old whose operations had taken place 7 to 78 months earlier (x = 35 months) and whose age at the time of surgery was a mean 7 years 6 months (range 60 to 144 months). Spirometry showed normal forced vital capacity for 7 of 8 patients (87 to 143% of theoretical value). Arterial oxygen saturation measured during and after a 6-minute walking test was normal for 7 of 8 patients. Chest films showed reduced lung volume on the affected side in 5 of 8 patients. In summary, lobectomy is a procedure with few complications. It requires a short hospital stay and has good postoperative prognosis. Function and x-ray studies show adequate lung growth and development, with normal lung function in those who could be so examined because they were old enough to cooperate. PMID- 11734138 TI - [Early discharge and home health care program for patients with exacerbated COPD and asthma]. AB - OBJECTIVE: To shorten hospital stays of patients with exacerbated chronic obstructive pulmonary disease (COPD) or asthma by way of a home care program and to assess whether the program increased the number of readmissions. METHODS: Patients admitted due to COPD exacerbation or asthma who did not need critical care and were discharged before the fourth day. A registered nurse experienced with respiratory disease patients and in regular contact with the pneumologist who supervised the program made follow-up home care visits to give instructions and check compliance with treatment. RESULTS: Sixty-nine patients enrolled in the program, 53 with COPD and 16 with asthma. A mean 7.2 home care visits per patient were made. The mean hospital stay was 3.69 ( 0.5 days for patients receiving home care and 7.89 ( 5 days for those who received no home care (p < 0.005). Severity of COPD in terms of age, FEV1 and PaO2 was similar in both groups, as follows: FEV1 was 39.4 12% and PaO2 66.3 7,7% for patients receiving home care; FEV1 was 40.6 ( 12% and PaO2 was 64.3 ( 7% for those receiving no home care (ns). The mean hospital stay overall for both groups was 7.4 (4.9 days; the mean hospital stay for the same diseases in the same previous the year before the study was 8.3 ( 5.5 (p < 0.05). The rate of readmissions for new exacerbations within 30 days of discharge was 4.3% (3/69) in the group receiving home care and 7.2% (29/401) among patients receiving only hospital care (ns.). A questionnaire survey at the end of the program showed satisfaction to be very high. CONCLUSIONS: A program of home care provided by a registered nurse experienced with respiratory diseases allows mean hospital stay to be reduced without increasing the number of readmissions within 30 days, with high patient satisfaction. PMID- 11734139 TI - [Intraoperative lymph node staging in bronchogenic carcinoma surgery. Consensus report]. AB - OBJECTIVES: To define methods for lymph node staging and the absence of nodal involvement (pN0) during surgery for bronchogenic carcinoma (BC). MATERIAL AND METHOD: Review of previous definitions of the Bronchogenic Carcinoma Cooperation Group of the Spanish Society of Pneumology and Chest Surgery (GCCB-S), and a comparison with the international literature; proposal of new definitions agreed upon by the GCCB-S. RESULTS: Methods for intraoperative lymph node evaluation proposed are as follows: not performed; biopsy (biopsy or removal of nodes with no intention of sampling all nodal stations); sampling (removal of nodes of a pre established number of nodal stations, including at least the paratracheal, subcarinal and hilar stations); systematic nodal dissection (SND) (excision of all nodes from stations on the operated side); extended nodal dissection (removal of contralateral pulmonary and mediastinal nodes as a complement to SND); and miscellaneous, for other forms of intraoperative staging. The proposed definition for pN0 is removal of at least six lymph nodes, pathology of all N1 locations, removal of lymph nodes from right superior and inferior paratracheal and subcarinal stations for right lung tumors, removal of subcarinal nodes for left lung tumors, removal of subaortic and anterior mediastinal nodes for tumors of the left upper lobe, and removal of paraesophageal and pulmonary ligament nodes for tumors of lower lobes. CONCLUSION: The new definitions encompass the current forms of intraoperative evaluation of BC and avoid the inaccuracies of previous definitions from the GCCB-S. PMID- 11734140 TI - [Present and future of thoracic surgery in Spain]. PMID- 11734141 TI - [Giant aneurysms of the pulmonary artery and peripheral venous thrombosis (Hughes Stovin syndrome): regression with immunosuppressant therapy]. AB - We report the case of a young Afro-Caribbean patient who complained of dry cough and low-grade fever. A chest film upon admission showed bilateral hilar masses in the lungs, which a CT scan with contrast medium confirmed were giant aneurysms of the pulmonary artery. The most relevant past history was deep venous thrombosis six months before admission. Hughes-Stovin syndrome was diagnosed and treatment was started with prednisolone and azathioprine. A scan 6 months after treatment ended showed the aneurysms had disappeared. Hughes-Stovin syndrome is characterized by pulmonary artery aneurysms and peripheral deep venous thromboses. Severe lung complications can include hemoptysis and thromboses inside the aneurysms. Hughes-Stovin syndrome has been considered a variety of BehCet's disease. PMID- 11734142 TI - [Pulmonary nocardiosis as a cause of radiographic imaging of multiple pulmonary nodules]. AB - Lung nocardiosis is a rare disease affecting patients with lymphoreticular neoplasm, immunodeficiency or chronic obstructive pulmonary disease; it can also affect patients who have received transplants. We report a case of lung nocardiosis in which radiographic presentation was acute, with a pattern of multiple bilateral pulmonary nodules ("cannonballs"), requiring us to rule out metastatic disease to arrive at a diagnosis. The patient responded slowly to antibiotic treatment until full resolution. PMID- 11734143 TI - [Fragmentation and fibrinolysis in pulmonary thromboembolism]. PMID- 11734144 TI - [Fibrinolysis and mechanical fragmentation in massive pulmonary embolism]. PMID- 11734145 TI - [The need for directly observed treatment in tuberculosis]. PMID- 11734146 TI - [Unusually located bronchial atresia]. PMID- 11734148 TI - [Vaccines: between new technologies and health policy]. PMID- 11734149 TI - [Peer-review: good, bad or quite the reverse?]. PMID- 11734150 TI - [Cancer incidence estimation in Spain: period 1993-1996]. AB - OBJECTIVE: To estimate the number of incident cases of cancer in Spain between 1993 and 1996. METHODS: We used data on the incidence of cancer from nine Spanish population-based cancer registries published in the monograph Cancer incidence in Five Continents, vols. VI and VII (period 1983-92). The National Institute of Statistics provided mortality data (period 1983-96). Generalized linear mixed models were used to estimate the number of incident cases. The incidence/mortality ratio was modeled. To account for heterogeneity, the providence of residence was considered as a random effect. Other factors analyzed were sex, age and period. Model parameters were estimated using a Bayesian approach with BUGS software. Estimates were valdated by comparing the observed number of cases with those predicted by the model in the regions with cancer registry data. RESULTS: The average number of incident cases per annum for all cancer sites except non-melanoma skin cancer was 78,440 for men and 55,480 for women. The most frequent neoplasm in men was lung with 15,480 cases followed by bladder with 9,445 cases and colorectal with 8,876 cases. In women the most frequent cancer was breast with 13,490 cases followed by colorectal with 8,274 cases. These absolute frequencies showed an increasing time trend for all cancers and for the most frequent sites, with the exception of stomach and uterine cervix. Internal validation of the estimates allowed calculation of a relative error smaller than 10%. IMPLICATIONS: This is the first time that the number of incident cases of cancer in Spain has been estimated with methods that account for heterogeneity. These figures show the importance of cancer as a public health problem in our community. PMID- 11734151 TI - [Space-time distribution of suicide in Galicia, Spain [1976-1998]]. AB - OBJECTIVE: To study the space-time distribution of suicide in Galicia from 1976 to 1998, taking into account gender, age, season and method of suicide. METHODS: For each sex crude and age-adjusted rates per province and age-specific rates per period (1978-84, 1985-91, 1992-98) were calculated. To estimate mortality risk and time trends in the municipalities, a Bayesian hierarchical model was used. RESULTS: In the last few years mortality from suicide has increase considerably. Age-adjusted rates of 6 per 105 at the beginning of the study rose to 10 per 105 at the end. The increase was greatest among men. The higher rates were observed in the elderly, although in terms of time trends the highest increase occurred among adolescents and young adults. In both sexes the main method of suicide was hanging. Less common methods were jumping from a height and shooting. The greatest number of suicides took place during the spring and summer months. In terms of spatial distribution, a high number of suicides were committed in the province of Lugo; in the municipalities, mortality risk decreased from North to South and an inverse temporal trend was observed in both sexes. This differed from the space-time patterns of suicides committed using methods other than hanging. CONCLUSIONS: Suicide is an important health problem in Galicia not only because to its magnitude but also because of the increasing rates, especially among young people. Geographical and temporal analysis can identify priority areas where preventive policies should be implemented. PMID- 11734152 TI - [Risk factors for cryptorchidism]. AB - OBJECTIVES: To identify the main risk factors for cryptorchidism in southeast Spain, and their possible association with environmental factors. METHODS: In this retrospective case-control study, the target population consisted of babies born in the University of Granada Hospital (UGH) in Granada, Spain, between 1 January 1992 and 31 December 1999, both inclusive. All boys from the target population aged 1 year or more diagnosed at the Pediatric Urology or Endocrinology Services of the UGH as having cryptorchidism (n = 70) were included, and a random sample of 144 baby boys born without cryptorchidism or any other related genitourinary abnormality or disease formed the control group. The mother's hospital record in the obstetrics clinic was checked to record information on potential risk factors, such as residence (specific health care district), parents' occupation, obstetric antecedents, diseases during pregnancy, gestational age at birth, birth weight and type of delivery. To search for associations between cryptorchidism and each of the variables we calculated crude and adjusted odds ratios with unconditional logistic regression analysis. RESULTS: Low gestational age at birth (37 weeks) and cesarian section were significantly associated with cryptorchidism. We also found an association with residence of the mother in the Costa-Alpujarra health care district, but its magnitude decreased when the association was adjusted for the father's occupation in agriculture. This last factor was also associated with cryptorchidism, although the association was not statistically significant. CONCLUSIONS: Our findings are consistent with those of other studies, except for the absence of association between low birth weight and cryptorchidism. Geographic variations in our study population suggested that environmental factors possibly related with farm work are involved in cryptorchidism. PMID- 11734153 TI - [Reliability and validity of self-reported drug use among secondary school students]. AB - OBJECTIVE: To assess the reliability and validity of self-reported use among secondary school students. METHOD: Validity was assessed in a representative sample of nearly 1,300 students by analyzing: a) the proportion of questions on drug use left unanswered compared with that for other questions; b) the proportion of inconsistencies between related questions; c) the proportion of questions wrongly completed; d) admision of ficticious drug use; e) the relationship between self-reported drug use and that of friends', and f) willingness to admit cannabis and ecstasy use. Reliability was analyzed using the kappa index, the proportion of specific agreement, and the intra-class correlation coefficient in a test-retest procedure in a randomized subsample of 349 students. RESULTS: The response rate to questions on drug use was high and was similar to that for more neutral questions. Only 0,3% of the secondary school students reported having used a fictitious drug. Except in the case of heroin, individual drug use was directly related to friends' perception of consumption. A very low proportion of students would not be willing to admit to use of cannabis (2%) or ecstasy (3,4%). Questions referring to drug use at some time during students' lives showed greater reliability than those referring to more recent drug use (last 12 months). The kappa indexes for drug consumption at some time during students lives ranged from 0.65 to 0.87, except for ecstasy and LSD (0.51 and 0.52 respectively). The age of first drug consumption was highly reliable (intra-class correlation coefficients ranged from 0.71 to 1). CONCLUSIONS: Except for ectasy, amphetamines and LSD, indexes of reliability and validity were generally good and similar to those obtained in other studies. These findings support the idea that information about self-reported drug use obtained through a questionnaire is reliable and valid, although the absolute prevalences of the use of some drugs should be interpreted with caution. The indicator of consumption at some time during students' lives is especially useful in studies monitoring drug use among this population. PMID- 11734154 TI - [Socio-economic costs of road traffic accidents in the Canary Islands, Spain, in 1997]. AB - OBJECTIVE: To evaluate the economic impact in terms of direct and indirect costs road traffic accidents in Canarias Islands (Spain) in 1997. METHOD: The cost-of illness method was used. Direct and indirect costs were estimated using prevalence cost, i.e., the costs produced in 1997. Direct costs were divided into health services costs, insurance administration costs and the costs of material damage to the vehicles. Indirect costs were obtained through transformation of physical units into monetary units using the approach of human capital theory. RESULTS: The total cost of road traffic accidents was 39,887.16 million pesetas, equivalent to 24,470 for each inhabitant of the Canary Islands and representing 1.3% of the GNP in this region. The total direct cost was 32,559.67 million pesetas, constituting 82% of the total, which was distributed according to the different concepts analyzed: health service costs: 2,407.40 million pesetas; insurance administration costs, 13,415.89 million pesetas and the costs of material damages to the vehicles: 16,736.38 million pesetas. The total indirect costs was 7,327.49 million pesetas, accounting for 18% of the total costs, which was distributed in premature mortality (6,884.88 million pesetas) and absenteeism from work (442.61 million pesetas). CONCLUSIONS: Although this study adopts a conservative approach by omitting costs associated with pain and suffering, permanent disability, and those of at-home care provided by the family, the hight socio-economic cost of road traffic accidents clearly indicates the need for the different administrations of the Canary Islands to collaborate in implementing preventive measures. PMID- 11734155 TI - Risk adjustment: beyond patient classification systems. AB - Diagnosis related groups (DRGs) are widely used in several countries. Their various versions aim to value the cost of hospital production. In Europe, the patient classification systems and standard weights used are usually the American originals. OBJECTIVES: The objective of this study was to analyse the extent to which DRGs and DRG-weights explain patient cost variability. Different components of patient cost (severity, comorbidities, complications and socioeconomic status), which are not well explained by DRG and which can be approximated by using administrative data, were also analysed. METHODS: A total of 35,262 discharges from two public hospitals in Barcelona were analysed. The Health Care Financing Administration (HCFA)-DRGs and the All Patient Refined (APR)- DRGs were calculated. Severity was adjusted by Disease Staging, and comorbidities and complications were calculated using Elixhauser and Charlson comorbidities groupings. An ecological socioeconomic status indicator was used. Linear regressions were estimated to explain per-patient cost variability. RESULTS: We found that Medicare's DRG-weights explained only 19% of cost variability. Cost based weights explained nearly 40% (38-42%, depending on the DRG classification used). Exclusion of outliers increased explanatory power to R2 = 47-48%. The remaining adjustment variables increased R2 to 49-51%. DISCUSSION: Medicare's DRG weights are not well-suited to Europe. Cost-based DRG-weights and outlier trimming have significantly greater explanatory power. The remaining clinical and socioeconomic variables have considerably less explanatory power but were statistically significant and behaved as expected. Spanish and other European health authorities should adapt DRG-classification systems to their environments for use in hospital production cost valuation. PMID- 11734156 TI - [Residents' assessment of education received in hospital departments: a monitoring tool]. AB - OBJECTIVE: [corrected] The Spanish postgraduate medical education system has made great contributions to the development of the National Health Service. Despite recent regulations on critical aspects of this education system, there still remains a need for a global assessment process. Hospital services evaluation by the residents should play a part in this process. METHODS: Administration over three years in a general teaching hospital of a specific questionnaire devoted to measure residents' perceptions of tthe medical education provided by their own service or department. Only residents who had stayed at least one year in their service were allowed to participate. Multivariable analyses using Multiple Correspondence Analysis (MCA) and Automatic Classification (AC) methods were performed. RESULTS: The overall response rate was 66.6% (325/488). 84,7% of the respondents deemed the overall education received as either adequate or excellent. The better scored aspects were patient management education, ethics education and residents' patient care supervision (more than 80% responses rating them as adequate/excellent). Performance of interdepartamental and bibliographic sessions along with research education were the worse perceived aspects by residents (less than 50% of responses as adequate/excellent). A factorial plane that explained 95% of overall response variability and allowed to rank the residents according to their assessment of education was obtained. Services with utmost ratings were found. AC results showed that were three different groups on the basis of overall peception of education received: The first group (18.2%) deemed it as excellent, the second group (61.5%) as adequate, while the thrid group (20.3%) considered it as inadequate. CONCLUSIONS: The administration of this questionnaire to hospital's residents and the analysis of its results using specific multivariable techniques provides useful information in order to monitor postgraduate medical education programmes and detect areas of improvement. PMID- 11734157 TI - [Identification of doctors with high pharmaceutical expenditure]. AB - OBJECTIVE: [corrected] To identify doctors with high pharmaceutical expenditure through multiple regression of variables not exclusively dependent on the doctor's decision. To compare the agreement with the method of the Spanish national health servicem which only considered the percentage of pensioners of the doctor's list. METHODS: Cross-sectional, retrospective, descriptive study of the pharmaceutical expenditure of 220 general practitioners in the health district of Toledo (Spain) in 1999. The following variables were collected: pharmaceutical expenditure for policy-holder/month (PEP-H); age; sex; habitat; size of the list; percentage of pensioners; frequency of attendance (consults/policy-holder-year) and workload (consultations per day). A multiple regression model with backwards elimination was constructed, taking like PEP-H as the dependent variable and the remaining as the independent variables. The resultant equation enabled calculation of the expected PEP-H for each doctor and the deviation of their real expenditure ofrom the expected. Doctors were considered to have high pharmaceutical expenditure when the deviation was more than the mean plus 1 standard deviation of this distribution. RESULTS: The mean PEP-H was 2,584.4 pesetas. Differences were found (F = 11.665; p < 0.005) in PEP H per habitat (2,723.2 in rural, 2,521.4 in semi urban and 2,168.2 in urban). A significant correlation was found (p < 0.005) between PEP-H and percentage of pensioners (r = 0.728) and frequency of attendance (r = 0.607). Our final model included percentage of pensioners, frequency of attendance, and age (F = 102.33; p < 0.005; r = 0.767; r2 = 0.588; beta = 206.05; beta1 = 48.27; beta2 = 61.26; beta3 = 9.55). This model were identified 25 DHPC. With the INSALUD model the identification were of 31. The Kappa index showed that agreement between both methods was 0.706 (SE 0.056) and simple disagreement was found in the classification of 24 doctors (10.9%). CONCLUSIONS: Frequency of attendance and the percentage of pensioners on doctors' lists are iclosely related to PEP-H. The indicator currently used by the Spanish national health system to identify doctors with high pharmaceutical expenditure could be improved if other variables, such as frenquency of attendance and the doctors' age, were taken into account. The adoption of this method would make budget allocation more impartial and fair. PMID- 11734159 TI - [The World Health Organization Report 2000: a politically incorrect computer game]. AB - The World Health Organization Report 2000 is aimed at supporting an evidence based development of health care systems in the world. The report has brought about a significant political and academic debate. This article reviews the contribution of the WHO Report to current health care policy, as well as analyzes and comments the main published critiques. Through a commented compilation of the health care reforms implemented over the last fifteen years in the world, WHO shifts radically its traditional position supporting now universal access to health care and a strong government's conduction role, but within a financial risk sharing environment, public and private co-operation and deregulation of the public management of health care institutions. WHO ranks in this report the 191 countries according to their health care systems' goal attainment and performance using a very weak, obscure and artificial methodology which has been opposed internationally. WHO is, however, committed to continue this initiative without apparently any major changes in the general and methodological approach. PMID- 11734158 TI - [Negative binomial distribution versus Poisson in the analysis of recurrent phenomena]. AB - OBJECTIVE: The aim is to unfold the difficulties likely to arise in risk calculations through aggregated database when the studied phenomenon is recurrent and to display the negative binomial distribution as a valid and simple alternative to analyse this kind of phenomenon. METHODS: When the studied phenomenon is recurrent, the analysis by means of the Poisson regression can provoke overdispersion or extra-poisson variance, what leads to underestimating the standard errors in coefficients and may divert into the statistical significance of factors which as a matter of fact are not associated with the phenomenon beforehand. The negative binomial can grasp part of the variance which the Poisson is unable to identify. In order to check this out, the fit of both distributions were compared, based on the number of hospitalizations of individuals aged between 65 and 69, during 1996. This comparison was carried out by means of two different aggregated databases: by individuals and by variables. RESULTS: There were differences in the fitted models by means of both distributions in both databases. By the analysis of the residuals, when using the base by individuals, the negative binomial fits correctly 67.9% of the observations badly fitted by the Poisson. Using the aggregated variables database, the percentage is 50%. In both cases, Poisson estimates four out of the six studied variables as significant. As to the negative binomial, there are two significant based on individuals and one in the variable database. CONCLUSION: The existence of overdispersion is frequent in recurrent-type phenomena. When this occurs, the negative binomial distribution is more appropiate than the Poisson. PMID- 11734160 TI - [Critique of conventional wisdom. The World Health Organization's report "WHO Health Systems: improving performance" and its relevance in Spain]. AB - This article provides a critical review of the World Health Organization's (WHO) report, WHO Health Systems: improving performance. The political context in which the report was presented, the ideological suppositions underlying the report's evaluation of the efficiency and efficacy of the health systems, their degree of responsiveness to users and the progressiveness of their funding are analyzed. Finally, the possible relevance of the report in Spain is discussed. PMID- 11734164 TI - [Health legislation and the challenges of the new century. Crimes against public health]. PMID- 11734165 TI - [Legionella: a problem for the Public Administration]. PMID- 11734168 TI - [Risk factors associated to hospitalization in diabetic patients with foot ulcers]. AB - BACKGROUND: To analyse the risk factors associated with admissions of diabetic patients with foot ulcers, in order to prevent hospitalization. In-patient care supposes the greatest cost of the diabetic foot care. SUBJECTS AND METHOD: We studied 108 diabetic patients (male n = 59, mean age 68 [11] years, 100 type 2 diabetics, disease duration 16.4 [10.3]) who attended the diabetic foot unit between January 1996 and September 1997. Clinical and biochemical risk parameters were recorded in a clinical protocol. Patients were followed up for a mean period of 200.2 (121.3) days. Admission was needed for 30 diabetic patients (28%). RESULTS: Risk factors for hospitalization were: higher ulcer diameter (1.9 [1.6] vs 3.4 [2.3] cm, p < 0.05), higher ulceration grade according to Wagner's classification (p = 0.005), presence of > 2 cm cellulitis (p < 0.001), worse metabolic control (HbA1c 7.6 [1.3] vs 8.6 [1.1], p < 0.05), and greater degree of retinopathy (p < 0.005). Odds ratios (OR) for admission were: advanced diabetic retinopathy: OR 1.9 (CI 95% 1.3-2.9; p = 0.02); presence of moderate to severe polyneuropathy: OR 1.6 (CI 95% 1.3-2.1; p = 0.05); upper HbA1c tertile: OR 2.4 (CI 95% 1.2-4.2; p = 0.004). In the logistic regression model, both foot ulcer diameter and advanced diabetic retinopathy were significantly independent. CONCLUSIONS: In diabetic subjects with foot ulcers, hospitalization is independently related to the diameter of the ulcer and the degree of diabetic retinopathy. Therapeutic and preventive approaches should be strengthened in this subgroup of diabetic patients to prevent their hospitalization. PMID- 11734169 TI - [The Spanish scientific production about human immunodeficiency virus infection/AIDS. A study through MedLine (1991-1999)]. AB - BACKGROUND: To analyze the research output of Spanish authors on human immunodeficiency virus (HIV) infection/AIDS. METHODS: Papers published over 1991 1999 which were recorder in the MedLine database (CD-ROM) were selected by using the key words Spain or Espana in the address of author field and HIV, AIDS, VIH or sida in any field. Papers were analyzed according to publication journal specialty of authors, subjecte matter of the paper, institution, center of origin and geographical distribution. RESULTS: A total of 2,065 papers were studied. Their number doubled, from 148 in 1991 to 295 in 1999 (r = 0.917; p < 0.0001). The number of papers published in foreign journals trebled, from 51 to 182 (r = 0.933; p < 0.001). The journal with the highest number o papers published was Medicina Clinica: 241 (11.7%). Although the main topic was the clinical manifestations of HIV infection (64.6%), the proportion of papers on this subject decreased from 71.4% in 1991-93 to 58.3% in 1997-99 (p < 0.001). On the other hand, the percentage of articles on antiretroviral therapy increased from 3.6% in 1991-93 to 12.4% in 1997-99 (p < 0.001). The greatest number of contributions came from hospitals (79.8% of papers), followed by universities (9.5%). Centers with the highest output were the Hospital Clinic (Barcelona) (5.7%) and the Hospital Carlos III (Madrid) (5.5%). The Autonomous Communities of Madrid (33.1%) and Catalonia (25.3%) concentrated more than half of the total output, followed by Andalusia (11.5%) and the Valencian Community (8.1%). CONCLUSIONS: Spanish output on HIV infection/AIDS research grew during the 1991-1999 period. There was a remarkable increase in the papers published in foreign journals. Moreover, topics changed according to the epidemiological changes observed in HIV infection during the past decade. PMID- 11734170 TI - [Clinical and epidemiological characteristics of persons newly diagnosed of HIV infection]. AB - BACKGROUND: We describe the characteristics of people with a recent diagnosis of HIV infection. PATIENTS AND METHOD: Series of 126 new cases of HIV infection diagnosed in an infectious diseases Service. RESULTS: Sexual transmission was the commonest form of infection (74%). Fifteen percent of individuals were aged over 50 years and 16% were immigrants. A late diagnosis (stage C) accounted for 36% of cases and it was associated with being more than 50 years old (OR: 5.1; 95% CI: 1.6-16.8). CONCLUSIONS: It is necessary to implement HIV infection surveillance systems and to improve preventive services. PMID- 11734171 TI - [Etiology of community-acquired pneumonia in ambulatory patients. Usefulness of a diagnostic investigation protocol using detection of Streptococcus pneumoniae and Legionella pneumophila antigens in urine samples]. AB - BACKGROUND: To determine the etiology of community-acquired pneumonia (CAP) in ambulatory patients and to assess the efficiency of a diagnostic protocol by using tests to detect bacterial antigens in urine samples. PATIENTS AND METHOD: One-year prospective study that included blood and sputum cultures, serologic studies, and detection of Legionella pneumophila and Streptococcus pneumoniae urinary antigens. RESULTS: 49 patients were recruited and an etiological diagnosis was attained in 34 (69%). Microorganisms most frequently isolated were S. pneumoniae (12 cases), Mycoplasma pneumoniae (7), Haemophilus influenzae (4), respiratory viruses (4) and Coxiella burnetii (3 cases). CONCLUSIONS: By means of a non-invasive protocol with urinary antigen tests, a microbial etiology can be established in two thirds of patients with mild CAP. S. pneumoniae is the main cause of mild CAP. PMID- 11734172 TI - [Gaucher's disease and other lisosomal diseases]. PMID- 11734173 TI - [Rational use of health care resources]. PMID- 11734174 TI - [Safety of quinolones administration in children?]. PMID- 11734175 TI - [Cigarette smoking and subjective health]. PMID- 11734176 TI - [Hepatitis C virus infection and genesis of lymphomas]. PMID- 11734177 TI - [Misuse of the term "fever of unknown origin"]. PMID- 11734178 TI - [Knowledge and skills and aptitudes in healthcare management]. PMID- 11734179 TI - [Knowledge and skills and aptitudes in healthcare management]. PMID- 11734181 TI - 5-HT receptor subtypes involved in the spinal antinociceptive effect of acetaminophen in rats. AB - The present study was designed to investigate which subtype of spinal 5-HT receptors were involved in acetaminophen-induced antinociception using the paw pressure test. Propacetamol (prodrug of acetaminophen, 400 mg/kg, injected intravenously, corresponding to 200 mg/kg of acetaminophen) produced a significant antinociceptive effect in this test. This effect was at least partially inhibited by intrathecal (i.t.) pretreatment with the 5-HT(1B) (penbutolol), 5-HT(2A) (ketanserin), 5-HT(2C) (mesulergine) receptor antagonists, but not by the 5-HT(1A) (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl)cyclohexanecarboxamide trihydrochloride, WAY 100635) and 5-HT(3) (granisetron) receptor antagonists. This profile was very close to that obtained recently with 5-HT, which suggests an implication of 5-HT in the spinal antinociceptive effect of acetaminophen. These results, the lack of binding of acetaminophen to 5-HT receptors and the increase of central 5-HT levels induced by this drug suggest that acetaminophen-induced antinociception could be indirectly mediated by 5-HT. PMID- 11734182 TI - In vitro activity of LY393558, an inhibitor of the 5-hydroxytryptamine transporter with 5-HT(1B/1D/2) receptor antagonist properties. AB - 1-[2-[4-(6-fluoro-1H-indol-3-yl)-3,6-dihydro-1(2H)-pyridinyl]ethyl]-3-isopropyl-6 (methylsulphonyl)-3,4-dihydro-1H-2,1,3-benzothiadiazine-2,2-dioxide (LY393558) is a potent inhibitor of [3H]5-hydroxytryptamine ([3H]5-HT) uptake into rat cortical synaptosomes (pIC(50)=8.48+/-0.12). It produces a dextral shift of the 5-HT dose response curves for the binding of GTPgamma[35S] to human 5-HT(1B) (pK(b)=9.05+/ 0.14) and 5-HT(1D) (pK(b)=8.98+/-0.07) receptors and inhibits the contractile response of the rabbit saphenous vein to the 5-HT(1B/D) receptor agonist, sumatriptan (pK(b)=8.4+/-0.2). In addition, it is an antagonist at the 5-HT(2A) (pK(i)=7.29+/-0.19) and 5-HT(2B) (pK(i)=7.35+/-0.11) receptors. Presynaptic autoreceptor antagonist activity was demonstrated by its ability to potentiate the K(+)-induced outflow of [3H]5-HT from guinea pig cortical slices (pEC(50)=7.74+/-0.05 nM) in which the 5-HT transporter had been inhibited by a maximally effective concentration of paroxetine. It is concluded that LY393558 should be an effective antidepressant with the potential to produce an earlier onset of efficacy than selective serotonin uptake inhibitors. PMID- 11734183 TI - LY393558, a 5-hydroxytryptamine reuptake inhibitor and 5-HT(1B/1D) receptor antagonist: effects on extracellular levels of 5-hydroxytryptamine in the guinea pig and rat. AB - The stimulation of terminal 5-HT(1B/1D) autoreceptors limits the effects of selective serotonin reuptake inhibitors on extracellular levels of 5 hydroxytryptamine (5-HT, serotonin) in vivo. Microdialysis studies show that acute oral administration of LY393558-a 5-HT reuptake inhibitor and antagonist at both the human 5-HT(1B) and 5-HT(1D) receptor-in the dose range 1-20 mg/kg, increases extracellular levels of 5-HT in both the guinea pig hypothalamus and rat frontal cortex. In both species, the levels of 5-HT that were attained were higher than following an acute, maximally effective dose of fluoxetine (20 mg/kg orally), reaching approximately 1500% in the guinea pig hypothalamus and 700% in the rat frontal cortex. In both species, the response to LY393558 (10 mg/kg p.o.) was impulse dependent, being absent in the presence of tetrodotoxin delivered at 1 microM via the microdialysis probe. The sensitivity to tetrodotoxin contrasted with the effects seen with DL-fenfluramine. Studies in rats showed that the microdialysate 5-HT concentration achieved in the frontal cortex after an acute challenge with LY393558 (5 mg/kg p.o.) was significantly greater than following a chronic regime of fluoxetine treatment (10 mg/kg/day orally for 21 days). Moreover, in rats chronically treated with LY393558 (5 mg/kg/day orally for 21 days), the mean basal concentration, 24 h after the final pretreatment dose, was of the same magnitude as that following chronic fluoxetine. However, in contrast to the response seen in fluoxetine-pretreated animals, a challenge dose of LY393558 still elicited a further increase in extracellular 5-HT in LY393558 pretreated animals. LY393558 is a potent 5-HT reuptake inhibitor and 5-HT(1B/1D) receptor antagonist. Microdialysis studies show that acute oral administration increases extracellular levels of 5-HT, by an impulse-dependent mechanism, above those produced by a maximally effective dose of fluoxetine, and in rats to levels only achieved following chronic fluoxetine treatment. Its neurochemical profile in vivo suggests that it may be a more effective antidepressant with the potential for producing an earlier onset of clinical activity than selective serotonin reuptake inhibitors. PMID- 11734184 TI - Catalepsy and hypolocomotion induced by a nitric oxide donor: attenuation by theophylline. AB - Nitric oxide (NO) promotes adenosine release in the striatum and hippocampus. Behavioral effects of the nitric oxide donor sodium nitroprusside were studied in mice and included an examination of spontaneous locomotion and catalepsy, which are behaviors modulated by adenosine. Sodium nitroprusside caused a dose dependent (2, 4 and 6 mg/kg) decrease in locomotor activity and catalepsy at the dose of 6 mg/kg. These effects were substantially attenuated by pretreatment with the non-selective adenosine receptors antagonist theophylline (10 and 30 mg/kg). Moreover, combined treatment with theophylline (30 mg/kg) and sodium nitroprusside (6 mg/kg) induced limbic seizures in 23% of animals. The pretreatment with the selective adenosine A(1) receptor antagonist 8-cyclopentyl 1, 3-dimethylxanthine (CPT) (1.2 mg/kg) caused no effect on the spontaneous or sodium nitroprusside-induced behavior. These data suggest that these behavioral effects of sodium nitroprusside are at least partially mediated by adenosine in the striatum and hippocampus, probably via adenosine A(2A) receptors. PMID- 11734185 TI - Lack of robust anticonvulsant effects of muscimol microinfusions in the anterior substantia nigra of kindled rats. AB - The substantia nigra pars reticulata is thought to control the spread of seizures in various seizure models. Potentiation of gamma-aminobutyrate (GABA)-mediated transmission in this region by intranigral administration of drugs such as muscimol has been shown to inhibit seizure propagation in such models, including the kindling model of epilepsy. More recent studies have shown that the effects on seizures are site-specific within the substantia nigra pars reticulata. Using flurothyl to induce clonic seizures, it was reported that bilateral microinfusions of muscimol into the anterior substantia nigra pars reticulata were anticonvulsant, while similar infusions into the posterior pars reticulata were proconvulsant. This prompted us to reevaluate the effects of intranigral muscimol in the kindling model with particular emphasis on the anterior substantia nigra pars reticulata. In amygdala kindled rats, muscimol was bilaterally infused into the anterior pars reticulata at doses of either 60 or 120 ng. Thirty minutes later, the threshold for induction of afterdischarges in the amygdala and the threshold for generalized seizures were determined in each rat. Furthermore, severity and duration of seizures at threshold currents were recorded. Unexpectedly, muscimol failed to increase seizure thresholds or to significantly reduce seizure severity or duration of motor seizures, although there was a moderate reduction in motor seizure duration in several rats. The data indicate that, in contrast to flurothyl seizures, in kindled rats the anterior pars reticulata of the substantia nigra is not a site at which muscimol causes robust anticonvulsant effects. PMID- 11734186 TI - Opposite effects of substance P fragments C (anxiogenic) and N (anxiolytic) injected into dorsal periaqueductal gray. AB - Recent findings implicating neurokinins in the expression of anxiety-like behaviors have stimulated interest in the participation of these neuropeptides in the dorsal periaqueductal gray matter (dPAG), one of the main output regions of the brainstem for the expression of defense reaction. Studies on the behavior of rats submitted to the elevated plus-maze test in this laboratory have shown that microinjections of substance P into the dorsal periaqueductal gray produce anxiogenic-like effects. Now, we analyze what portion of the molecule of substance P is responsible for these effects through the examination of the action of its C- and N-terminus fragments (6-11 and 1-7) in the elevated plus maze. We also investigated whether these effects are influenced by prior treatment with the tachykinin NK(1) receptor antagonist 17-beta-hydroxy-17-alpha ethynyl-5alpha-androstanol[3,2-b]pyrimido[1,2-a]benzimidazole (WIN51,708). To this end, rats were implanted with a cannula in the dorsal periaqueductal gray and injected 1 week later with equimolar doses (17.5 and 35 pmol) of either C- or N-fragments of substance P and tested in the elevated plus-maze. The results show that the C-terminal fragment has an anxiogenic profile of effects, including reduction in the number of entries and time spent in the open arms of the maze, plus increases in scanning, stretched-attend posture, head dipping and flat-back approach. On the other hand, the N-terminal fragment produced opposite effects, namely, an increase in the number of entries and time spent in the open arms of the maze accompanied by an increase in end-arm activity, rearing and head dipping. The tachykinin NK(1) receptor antagonist WIN51,708 (20 mg/kg, i.p.) inhibited the effects of the carboxy-terminal of substance P while it did not change the effects of the N-terminal fragment. Microinjection of WIN51,708 (20 mg/kg, i.p.), by its own, did not produce any significant effects. Therefore, the results indicate that the anxiogenic effects of substance P injected into the dorsal periaqueductal gray are encoded by its carboxy-terminal sequence and due to its action on tachykinin NK(1) receptors. PMID- 11734187 TI - Effects of CP-154,526 on responding during extinction from cocaine self administration in rats. AB - Conditioned cues associated with cocaine induce craving and relapse. Although the role of corticotropin releasing hormone (CRH) in stress- and cocaine-induced relapse has been reported, its involvement in cue-induced behavior has not been established. Using responding during extinction as a model of cue-induced craving, we tested the effects of a selective CRH1 receptor antagonist, CP 154,526 (butyl-ethyl-[2,5-dimethyl-7-(2,4,6-trimethyl-phenyl)-7H-pyrrolo[2,3 d]pyrimidin-4-yl]-amine). Rats were trained to respond on a multiple schedule of cocaine self-administration and food reinforcement. On extinction test days, saline was substituted for cocaine. Pretreatment with CP-154,526 (20 mg/kg, i.p.) decreased responding on the cocaine-associated lever during extinction, suggesting an involvement of CRH1 receptors in cue-induced craving. PMID- 11734188 TI - Effects of neurotensin receptor activation on brain stimulation reward in Fischer 344 and Lewis rats. AB - The effects of intracerebroventricular injections of 18 nmol/10 microl of neurotensin, [D-Tyr(11)]neurotensin, or saline on operant responding for brain stimulation reward were investigated in Fischer 344 (F344) and Lewis (LEW) rats using the curve-shift paradigm. [D-Tyr(11)]neurotensin, but not neurotensin, decreased reward threshold in F344 rats while it increased thresholds in LEW rats. Both peptides suppressed maximal rates of responding; this effect was of greater magnitude and longer lasting in LEW than in F344 rats. These findings show that F344 and LEW rat strains are differentially sensitive to activation of central neurotensin receptors that modulate reward-relevant circuitry. PMID- 11734189 TI - Novel marine-derived halogen-containing gramine analogues induce vasorelaxation in isolated rat aorta. AB - We examined the effects of 2,5,6-tribromo-1-methylgramine (TBG), isolated from bryozoan, and its derivative, 5,6-dibromo-1,2-dimethylgramine (DBG), on the contraction of rat aorta. TBG and DBG decreased the high-K(+)-induced increase in muscle contraction and cytosolic Ca(2+) level ([Ca(2+)](i)), respectively. The inhibitory effects of TBG and DBG on high-K(+)-induced contraction were antagonized by increasing the external Ca(2+) concentration or by 1,4-dihydro2,6 dimethyl-5-nitro-4-[2-(trifluoromethyl)phenyl]pyridine-3-carboxylic acid (Bay k8644). The high-K(+)-induced increase of Mn(2+) influx was completely blocked by 10 microM TBG or 10 microM DBG. In the Ca(2+)-free solution, 30 microM TBG or 30 microM DBG inhibited the phenylephrine-induced transient increase in [Ca(2+)](i) and muscle tension, while scarcely affecting caffeine-induced transient changes. TBG and DBG significantly increased the cyclic AMP content at 30 microM, but not at 10 microM. These results suggest that TBG and DBG inhibit the smooth muscle contraction by inhibiting Ca(2+) entry, and at higher concentrations, the increase in intracellular cyclic AMP content also contributes to their inhibitory effect. PMID- 11734190 TI - Anti-arrhythmic and electrophysiological effects of the endothelin receptor antagonists, BQ-123 and PD161721. AB - The effects of the endothelin ET(A), (BQ-123) and endothelin ET(A/B) (PD161721) receptor antagonists were investigated on ischaemia-induced arrhythmias and on the maximum following frequency. The study was carried out in Langendorff perfused rat hearts subjected to coronary artery occlusion in which the severity of arrhythmias, coronary perfusion pressure and heart rate were measured. The % incidence of ischaemia-induced irreversible ventricular fibrillation (ventricular fibrillation) was reduced significantly from 58%, in control rat hearts, to 0% (at 10(-7) and 10(-6) M PD161721 and 10(-6) M BQ-123 P<0.05). Maximum following frequency was measured in guinea-pig isolated atria. In the presence of normal extracellular [K(+)], BQ-123 and PD161721, at 10(-6) M, significantly decreased the maximum following frequency from 9.0+/-0.7 to 7.2+/-0.4 and from 8.3+/-0.4 to 6.7+/-0.3 Hz, respectively (P<0.05). These effects were not potentiated by raising the extracellular [K(+)] with the exception of 10(-9) M PD161721. In contrast, lignocaine's ability to reduce the maximum following frequency was greater in elevated (e.g. at 1.7x10(-4) M from 8.4+/-0.3 to 2.5+/-0.6 Hz) than in normal [K(+)] (from 9.0+/-0.3 to 4.9+/-0.5 Hz). In conclusion, both BQ-123 and PD161721 had an anti-fibrillatory effect in isolated rat hearts that may be due, at least in part, to an ability to reduce the maximum following frequency. This latter effect is unlikely to be due to Na(+) channel blockade since it was not markedly potentiated by elevation of extracellular [K(+)]. PMID- 11734191 TI - Protective effects of M40403, a superoxide dismutase mimetic, in a rodent model of colitis. AB - Inflammatory bowel disease is characterised by oxidative and nitrosative stress, leukocyte infiltration, and up-regulation of intercellular adhesion molecule 1 (ICAM-1) expression in the colon. The aim of the present study was to examine the effects of M40403, a superoxide dismutase mimetic, in rats subjected to experimental colitis. Colitis was induced in rats by intracolonic instillation of trinitrobenzene sulfonic acid (TNBS). Rats experienced bloody diarrhoea and significant loss of body weight. At 4 days after TNBS administration, the colon damage was characterised by areas of mucosal necrosis. Neutrophil infiltration (indicated by myeloperoxidase activity in the mucosa) was associated with up regulation of ICAM-1 and expression of P-selectin and high levels of malondialdehyde. Immunohistochemistry for nitrotyrosine and poly (ADP-ribose) synthetase showed an intense staining in the inflamed colon. Treatment with M40403 (5 mg/kg daily i.p.) significantly reduced the appearance of diarrhoea and the loss of body weight. This was associated with a remarkable amelioration of the disruption of the colonic architecture as well as a significant reduction of colonic myeloperoxidase activity and malondialdehyde levels. M40403 also reduced the appearance of nitrotyrosine and poly (ADP-ribose) synthetase immunoreactivity in the colon as well as reduced the up-regulation of ICAM-1 and the expression of P-selectin. The results of this study suggested that administration of a superoxide dismutase mimetic may be beneficial for treatment of inflammatory bowel disease. PMID- 11734192 TI - Impairment by lovastatin of neural relaxation of the rabbit sphincter of Oddi. AB - We sought whether inhibition of cholesterol biosynthesis by lovastatin influenced the nitrergic relaxation response of the sphincter of Oddi. Rabbit sphincters of Oddi rings were tested for changes in isometric tension in response to field stimulation in the presence of 4 microM guanethidine and 1 microM atropine. Tissue samples were then analyzed for cAMP and cGMP content by radioimmunoassay for nitric oxide concentration by electron spin resonance and for vasoactive intestinal peptide and calcitonin gene-related peptide (CGRP) release by radioimmunoassay. Membrane G(salpha) protein was determined by Western blot analysis. Field stimulation relaxed the preparations with an increase in nitric oxide, cAMP and cGMP concentrations at increased calcitonin gene-related peptide and vasoactive intestinal polypeptide (VIP) release. Preparations from rabbits pre-treated with lovastatin (5 mg/kg/day intragastrically, over 5 days) contracted under the same conditions with an attenuated cGMP-increase at preserved increase in NO content and neuropeptide release. The relaxation was recaptured combining lovastatin with farnesol (1 mg/kg intravenously, twice a day for 5 days). The field stimulation-induced increase in cyclic nucleotides was also restored. Lovastatin decreased membrane G(salpha) protein content, which was re-normalized by farnesol. Farnesol treatment reinstates neurogenic relaxation of the sphincter of Oddi deteriorated by lovastatin possibly by normalizing G protein coupling. PMID- 11734193 TI - Endotoxin impairs biliary transport of sparfloxacin and its glucuronide in rats. AB - The effect of endotoxin on glucuronidation and hepatobiliary transport of quinolone antimicrobial agents was investigated in rats using sparfloxacin and p nitrophenyl glucuronide as model drugs. The biliary clearance experiments were performed 24 h after a single intraperitoneal injection of endotoxin (1 mg/kg). Endotoxin significantly delayed the disappearance of sparfloxacin from plasma and increased plasma concentration of its glucuronide after intravenous injection of sparfloxacin (10 mg/kg). Significant decreases in the systemic clearance of sparfloxacin and the biliary clearance of sparfloxacin and the glucuronide were observed. Endotoxin had no effect on in vitro glucuronidation activity using p nitrophenol as a substrate. When p-nitrophenyl glucuronide (8 mg/kg) was administered in endotoxin-pretreated rats, significant decreases in the systemic clearance, biliary clearance and renal clearance of p-nitrophenyl glucuronide were observed. These findings suggest that endotoxin decreases the biliary excretion of sparfloxacin and its glucuronide probably due to impairment of their hepatobiliary transport systems and renal handling. PMID- 11734194 TI - The anti-inflammatory activities of cannabinoid receptor ligands in mouse peritonitis models. AB - In this report, we describe experiments in which cannabinoid receptor ligands were evaluated for effects on the development of a peritoneal inflammation when elicited in mice with thioglycollate broth or staphylococcus enterotoxin A. The cannabinoid receptor agonists [(-)-11-hydoxy-Delta(8) tetrahydrocannabinol dimethylheptyl] (HU-210) and [(R)-(+)-[2,3-dihydro-5-methyl-3-[(4 morpholinyl)methyl[pyrrolo[1,2,3-de]1,4-benzoxazin-6-yl](1-naphthalenyl) methanone] (WIN 55212-2) blocked the migration of neutrophils into the peritoneal cavity in response to these inflammatory stimuli. This effect was caused by a delay in the production of the neutrophil chemoattractants, KC and macrophage inflammatory protein-2. HU-210 and WIN 55212-2 blocked neutrophil chemokines and neutrophil migration whether administered subcutaneously (s.c.) or intracerebroventricularly (i.c.v.). Their modulatory effects on the inflammation were antagonized by centrally administered [N-(piperdin-1-yl)-5-(4-chloropheny)-1 (2,4-dichloropheny)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride] (SR141716A), a selective cannabinoid CB(1) receptor antagonist. This latter observation, and the ability of the cannabinoid receptor agonists to suppress the peritoneal inflammation at relatively low doses when administered i.c.v., indicated a role for central cannabinoid CB(1) receptors in the anti-inflammatory activities of HU-210 and WIN 55212-2. The cannabinoid receptor agonists had no effect on monocyte migration elicited by thioglycollate, despite their ability to suppress monocyte chemotactic protein-1 levels in lavage fluids. The cannabinoid CB(2) receptor antagonist, [N-[(1S)-endo-1,3,3-trimethylbicyclo[2.2.1]heptan-2 yl]5-(4-choro-3 methylphenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide] (SR144528) inhibited the peritoneal inflammation in a manner analogous to that of HU-210 and WIN 55212-2 when administered i.c.v., but it did not appear to act through central cannabinoid CB(1) receptors. The present results add to the body of literature indicating that cannabinoid receptor ligands have diverse anti inflammatory properties. PMID- 11734195 TI - Ferredoxins of the third kind. AB - Most low-potential ferredoxins (Fds) are of the well-known [2Fe-2S] plant or [4Fe 4S] bacterial type. Yet, an additional class of [2Fe-2S] Fds has been recognized on the basis of sequence and spectroscopic idiosyncrasies. A recent crystal structure has confirmed the uniqueness of this third kind of Fd, and shown that these proteins display an unexpected structural similarity to thioredoxin. The properties of these thioredoxin-like [2Fe-2S] Fds are summarized, and hypotheses concerning their function are discussed. PMID- 11734196 TI - Identification of a new human lens UV filter compound. AB - A new UV filter compound, 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid O diglucoside, has been identified in human lenses. The structure suggests that it is a further metabolic product of the second most abundant UV filter compound, 4 (2-amino-3-hydroxyphenyl)-4-oxobutanoic acid O-glucoside. Quantification studies on the new compound show that it decreases towards zero in both the nucleus and cortex as a function of age. The discovery of this novel disaccharide completes the identification of the major UV filter compounds present in the human lens. PMID- 11734197 TI - Paternally inherited transgenes are down-regulated but retain low activity during early embryogenesis in Arabidopsis. AB - We investigated the timing of transgene activation after fertilisation in Arabidopsis following crosses and using two transgenic promoters (from the AtCYCB1 and AtLTP1 genes). Using both a transactivation system and direct transcriptional fusion to drive beta-glucuronidase reporter expression, reciprocal crosses showed a lack of expression of the paternal components. This is consistent with a lack of paternal genome activity previously reported during early seed development in Arabidopsis [Viella-Calzada et al. (2000) Nature 404, 91-94]. However, transactivation experiments of the BARNASE gene gave evidence that at least some paternal loci retain transcriptional activity, though at a low level, during early embryogenesis. PMID- 11734198 TI - Ca(2+)-induced folding of a family I.3 lipase with repetitive Ca(2+) binding motifs at the C-terminus. AB - In order to understand a role of the Ca(2+) ion on the structure and function of a Ca(2+)-dependent family I.3 lipase from Pseudomonas sp. MIS38, apo-PML, holo PML, holo-PML*, and the N-terminal domain alone (N-fragment) were prepared and biochemically characterized. Apo-PML and holo-PML represent refolded proteins in the absence and presence of the Ca(2+) ion, respectively. Holo-PML* represents a holo-PML dialyzed against 20 mM Tris-HCl (pH 7.5). The results suggest that the C terminal domain of PML is almost fully unfolded in the apo-form and its folding is induced by Ca(2+) binding. The folding of this C-terminal domain may be required to make a conformation of the N-terminal catalytic domain functional. PMID- 11734199 TI - alpha-synuclein metabolism and aggregation is linked to ubiquitin-independent degradation by the proteasome. AB - alpha-Synuclein has been implicated in the pathogenesis of Parkinson's disease based on mutations in familial cases of the disease and its presence in Lewy bodies. Here we show that over-expression of wild-type human alpha-synuclein is sufficient to induce inclusion formation in SH-SY5Y cells. In this cellular model, proteasome inhibition leads to an increase of alpha-synuclein accumulation in vivo without ubiquitylation. In accordance, we find that in vitro, unmodified alpha-synuclein can be directly degraded by the 20S proteasome. These findings suggest an ubiquitin-independent mechanism of proteasomal degradation for alpha synuclein and other natively unfolded proteins. PMID- 11734200 TI - A lipid transfer protein binds to a receptor involved in the control of plant defence responses. AB - Lipid transfer proteins (LTPs) and elicitins are both able to load and transfer lipidic molecules and share some structural and functional properties. While elicitins are known as elicitors of plant defence mechanisms, the biological function of LTP is still an enigma. We show that a wheat LTP1 binds with high affinity sites. Binding and in vivo competition experiments point out that these binding sites are common to LTP1 and elicitins and confirm that they are the biological receptors of elicitins. A mathematical analysis suggests that these receptors could be represented by an allosteric model corresponding to an oligomeric structure with four identical subunits. PMID- 11734201 TI - Trimethylamine-N-oxide-induced folding of alpha-synuclein. AB - The effect of the natural osmolyte trimethylamine-N-oxide (TMAO) on the structural properties and fibril formation of the natively unfolded protein human alpha-synuclein was studied using several physico-chemical methods. TMAO induced folding of alpha-synuclein: at moderate concentrations, a partially folded intermediate with enhanced propensity for fibrillation accumulated; at higher concentrations, alpha-synuclein was tightly folded and underwent self-association to form oligomers. The latter conformation was significantly helical and probably represents the physiologically folded form of the protein. PMID- 11734202 TI - A novel second myostatin gene is present in teleost fish. AB - We report on the isolation and characterisation of the complete cDNA sequence encoding a novel bone morphogenetic protein-like protein (sbMSTN-b) in the teleost fish Sparus aurata. The encoded protein is 68% identical to S. aurata MSTN at the amino acid level, and homologues were also found in Umbrina cirrosa and Tetraodon nigroviridis. Phylogenetic analysis suggests that the MSTN-b gene may be present in most, perhaps all, teleost fish species. RT-PCR on different tissues/stages indicates that MSTN-b is expressed almost exclusively in the central nervous system, starting from late larval stages. Quantitative analyses indicate an increase of sbMSTN-b expression in the brain associated with metamorphosis, at the same time as completion of nervous system differentiation. PMID- 11734203 TI - N-Acetylneuraminic acid uptake in Pasteurella (Mannheimia) haemolytica A2 occurs by an inducible and specific transport system. AB - The N-acetylneuraminic acid (NeuAc) transport system of Pasteurella (Mannheimia) haemolytica A2 was studied when this bacterium was grown in both complex and chemically defined media. Kinetic measurements were carried out at 37 degrees C in 50 mM Tris-HCl buffer, pH 8.0, containing 50 microg/ml bovine serum albumin. Under these conditions, the uptake rate was linear for at least 3 min and the calculated K(m) for NeuAc was 0.1 microM. The transport rate was increased by the addition of several cations and was inhibited by sodium arsenite (95%), N,N' dicyclohexyl-carbodiimide (50%), and 2,4-dinitrophenol (40%) at final concentration of 1 mM (each). These results support the notion that NeuAc uptake is an active sugar cation symporter. Study of specificities showed that glucosamine, mannose and mannosamine inhibited the transport of NeuAc in this bacterium. Analysis of expression revealed that the NeuAc transport system was induced by NeuAc and by the simultaneous presence of glucose and galactose in the growth medium. PMID- 11734204 TI - Overproduced p73alpha activates a minimal promoter through a mechanism independent of its transcriptional activity. AB - p73, the gene for a protein related to the tumor suppressor p53, encodes several variants which bear distinct carboxy-terminal structures as a result of alternative splicing. We and others showed that these splicing variants have different transcriptional effects on promoters with a p53-binding consensus sequence (p53BCS). Here we show that when transiently overexpressed, p73alpha but not p73beta activated several minimal promoters without the p53BCS, while p73gamma and p73epsilon activated them to a much lesser extent than p73alpha, and p53 suppressed the promoters without p53BCS as reported previously. Moreover, the results of RNase protection and RNA transfection assays suggested that this activation occurred at the transcriptional level. Deletion analysis of p73alpha revealed that the transactivation domain of p73 was not involved in this activity and the C-terminal region of p73alpha which is a specific structure of this variant was essential, suggesting that this phenomenon occurs independent of the transactivation activity of p73alpha and that the C-terminal extension of p73alpha may affect the basal level of transcription. PMID- 11734205 TI - Both N-terminal catalytic and C-terminal RNA binding domain contribute to substrate specificity and cleavage site selection of RNase III. AB - The double-stranded RNA-specific endoribonuclease III (RNase III) of bacteria consists of an N-terminal nuclease domain and a double-stranded RNA binding domain (dsRBD) at the C-terminus. Analysis of two hybrid proteins consisting of the N-terminal half of Escherichia coli RNase III fused to the dsRBD of the Rhodobacter capsulatus enzyme and vice versa reveals that both domains in combination with the particular substrate determine substrate specificity and cleavage site selection. Extension of the spacer between the two domains of the E. coli enzyme from nine to 20 amino acids did not affect cleavage site selection. PMID- 11734206 TI - A role for N-WASP in invasin-promoted internalisation. AB - Phagocytosis of Yersinia pseudotuberculosis occurs through interaction of the bacterial protein invasin with beta1-integrins. Here we report that N-WASP plays a role in internalisation of an invasin-expressing, avirulent strain of Y. pseudotuberculosis. Ectopic expression of N-WASP mutants, which affect recruitment of the Arp2/3 complex to the phagosome, reduces uptake of Yersinia. In addition, expression of the Cdc42/Rac-binding (CRIB) region of N-WASP has an inhibitory effect on uptake. Using GFP-tagged Rho GTPase mutants, we provide evidence that Rac1, but not Cdc42, is important for internalisation. Furthermore, activated Rac1 rescues Toxin B, CRIB and Src family kinase inhibitor PP2-mediated impairment of uptake. Our observations indicate that invasin-mediated phagocytosis occurs via a Src and WASP family-dependent mechanism(s), involving the Arp2/3 complex and Rac, but does not require Cdc42. PMID- 11734207 TI - Human monkeypox and smallpox viruses: genomic comparison. AB - Monkeypox virus (MPV) causes a human disease which resembles smallpox but with a lower person-to-person transmission rate. To determine the genetic relationship between the orthopoxviruses causing these two diseases, we sequenced the 197-kb genome of MPV isolated from a patient during a large human monkeypox outbreak in Zaire in 1996. The nucleotide sequence within the central region of the MPV genome, which encodes essential enzymes and structural proteins, was 96.3% identical with that of variola (smallpox) virus (VAR). In contrast, there were considerable differences between MPV and VAR in the regions encoding virulence and host-range factors near the ends of the genome. Our data indicate that MPV is not the direct ancestor of VAR and is unlikely to naturally acquire all properties of VAR. PMID- 11734208 TI - Targeting influenza virosomes to ovarian carcinoma cells. AB - Reconstituted influenza virus envelopes (virosomes) containing the viral hemagglutinin (HA) have attracted attention as delivery vesicles for cytosolic drug delivery as they possess membrane fusion activity. Here, we show that influenza virosomes can be targeted towards ovarian carcinoma cells (OVCAR-3) with preservation of fusion activity. This was achieved by incorporating poly(ethylene glycol) (PEG)-derivatized lipids into the virosome membrane. This PEG layer serves as shield to prevent interaction of HA with ubiquitous sialic acid residues and as spatial anchor for antibody attachment. Coupling of Fab' fragments of mAb 323/A3 (anti-epithelial glycoprotein-2) to the distal ends of PEG lipids resulted in specific binding of virosomes to OVCAR-3 cells. These antibody-redirected virosomes fused with membranes of OVCAR-3 cells in a pH dependent fashion. PMID- 11734209 TI - Activity of human Delta5 and Delta6 desaturases on multiple n-3 and n-6 polyunsaturated fatty acids. AB - Yeast co-expressing human elongase and desaturase genes were used to investigate whether the same desaturase gene encodes an enzyme able to desaturate n-3 and n-6 fatty acids with the same or different carbon chain length. The results clearly demonstrated that a single human Delta5 desaturase is active on 20:3n-6 and 20:4n 3. Endogenous Delta6 desaturase substrates were generated by providing to the yeast radiolabelled 20:4n-6 or 20:5n-3 which, through two sequential elongations, produced 24:4n-6 and 24:5n-3, respectively. Overall, our data suggest that a single human Delta6 desaturase is active on 18:2n-6, 18:3n-3, 24:4n-6 and 24:5n 3. PMID- 11734210 TI - Tubulin and microtubule are potential targets for brain hexokinase binding. AB - The metabolite-modulated association of a fraction of hexokinase to mitochondria in brain is well documented, however, the involvement of other non-mitochondrial components in the binding of the hexokinase is controversial. Now we present evidence that the hexokinase binds both tubulin and microtubules in brain in vitro systems. The interaction of tubulin with purified bovine brain hexokinase was characterized by displacement enzyme-linked immunosorbent assay using specific anti-brain hexokinase serum (IC(50)=4.0+/-1.4 microM). This value virtually was not affected by specific ligands such as ATP or glucose 6 phosphate. Microtubule-bound hexokinase obtained in reconstituted systems using microtubule and purified hexokinase or brain extract was visualized by transmission and immunoelectron microscopy on the surface of tubules. The association of purified bovine brain hexokinase with either tubulin or microtubules caused about 30% increase in the activity of the enzyme. This activation was also observed in brain, but not in muscle cell-free extract. The possible physiological relevance of the multiple heteroassociation of brain hexokinase is discussed. PMID- 11734211 TI - A novel dicationic polyamide ligand binds in the DNA minor groove as a dimer. AB - We have investigated DNA binding properties of a dicationic polyamide molecule (GL020924) that has exhibited unique protein displacement and gene regulation activities. Fluorescence, thermal melting and electrospray ionization mass spectrometry experiments showed that the binding stoichiometry of GL020924 is 2:1 to various DNA oligomers with 8-11 contiguous A/T bp. In accordance with those findings, circular dichroism experiments showed GL020924 binds as a partially staggered side-by-side dimer spanning 10-12 bp. These observations and molecular modeling studies demonstrate that the 2:1 GL020924-DNA complex may exhibit a novel form of stacking orientation involving at least partially parallel peptide groups. PMID- 11734212 TI - Crystal structure of a cyclic form of bovine pancreatic trypsin inhibitor. AB - The crystal structure of a cyclic form of a mutant of bovine pancreatic trypsin inhibitor has been solved at 1.0 A resolution. The protein was synthesized by native chemical ligation and its structure is almost indistinguishable from the previously described recombinant form of the same mutant; however, the new loop containing the former termini became much better ordered. PMID- 11734213 TI - Overexpression of hepatocyte growth factor/scatter factor promotes vascularization and granulation tissue formation in vivo. AB - The effect of hepatocyte growth factor/scatter factor (HGF/SF) during wound healing in the skin was investigated, using HGF/SF-overexpressing transgenic mouse model. Histological analysis of HGF/SF transgenic mouse excisional wound sites revealed increased granulation tissue with marked vascularization. Northern blot analysis demonstrated that, relative to control, vascular endothelial growth factor (VEGF) expression in transgenic skin was significantly higher at baseline and was robustly up-regulated during wound healing. Elevated levels of VEGF protein were detected immunohistochemically, predominantly in endothelial cells and fibroblasts within the granulation tissue of HGF/SF transgenic skin. Serum levels of VEGF were also elevated in HGF/SF transgenic mice. Thus, results from our study suggest that HGF/SF has a significant effect on vascularization and granulation tissue formation during wound healing in vivo, involving with induction of VEGF. PMID- 11734214 TI - Manganese superoxide dismutase induction by iron is impaired in Friedreich ataxia cells. AB - Iron-mediated oxidative stress has been implicated in the pathology of the neurodegenerative disease Friedreich ataxia (FRDA). Here, we show that normal upregulation of the stress defense protein manganese superoxide dismutase (MnSOD) fails to occur in FRDA fibroblasts exposed to iron. This impaired induction was observed at iron levels in which increased activation of the redox-sensitive factor NF-kappaB was absent. Furthermore, MnSOD induction could only be partially suppressed by antioxidants. We conclude that an NF-kappaB-independent pathway that may not require free radical signaling is responsible for the reduction of MnSOD induction. This impairment could constitute both a novel defense mechanism against iron-mediated oxidative stress in cells with mitochondrial iron overload and conversely, an alternative source of free radicals that could contribute to the disease pathology. PMID- 11734215 TI - Adenovirus-mediated preproinsulin gene transfer into adipose tissues ameliorates hyperglycemia in obese diabetic KKA(y) mice. AB - We investigated whether adenovirus-mediated preproinsulin gene transfer into insulin target tissues (adipocytes) ameliorates hyperglycemia in diabetic mice. KKA(y) mice, a genetically obese type 2 diabetic animal model, were treated with a single subcutaneous injection of recombinant adenovirus, Adex1CA-human preproinsulin (Adex1CA-pchi), into the epididymal fat pads. pchi mRNA was expressed only in adipose tissue in which mature insulin was produced. Three days after virus injection these mice showed a marked decrease of blood glucose levels (from about 400 to 200 mg/dl), and an intraperitoneal glucose tolerance test revealed the markedly improved glucose tolerance. There was no significant difference in serum insulin levels between control and recombinant adenovirus treated KKA(y) mice. The normalized glucose levels in diabetic mice were maintained for at least 2 weeks after the virus injection. This strategy could provide a novel and, most importantly, a simple and convenient gene therapy for obese type 2 diabetes patients. PMID- 11734216 TI - Engineering of the H2O2-binding pocket region of a recombinant manganese peroxidase to be resistant to H2O2. AB - The manganese peroxidase produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+), is easily inactivated by the hydrogen peroxide (H2O2) presented in the reaction. We attempted to increase H2O2 resistance by the conformational stabilization around the H2O2-binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non-oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H2O2 where the wild-type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non-bulky and non-oxidizable Ser. PMID- 11734217 TI - Effect of D609 on phosphatidylcholine metabolism in the nuclei of LA-N-1 neuroblastoma cells: a key role for diacylglycerol. AB - In our previous studies, TPA treatment of LA-N-1 cells stimulated the production of diacylglycerol in nuclei, probably through the activation of a phospholipase C. Stimulation of the synthesis of nuclear phosphatidylcholine by the activation of CTP:phosphocholine cytidylyltransferase was also observed. The present data show that both effects were inhibited by the pretreatment of the cells with D609, a selective phosphatidylcholine-phospholipase C inhibitor, indicating that the diacylglycerol produced through the hydrolysis of phosphatidylcholine in the nuclei is reutilized for the synthesis of nuclear phosphatidylcholine and is required for the activation of CTP:phosphocholine cytidylyltransferase. PMID- 11734218 TI - Identification of mouse trp homologs and lipid rafts from spermatogenic cells and sperm. AB - Intracellular Ca(2+) has an important regulatory role in the control of sperm motility, capacitation, and the acrosome reaction (AR). However, little is known about the molecular identity of the membrane systems that regulate Ca(2+) in sperm. In this report, we provide evidence for the expression of seven Drosophila transient receptor potential homolog genes (trp1-7) and three of their protein products (Trp1, Trp3 and Trp6) in mouse sperm. Allegedly some trps encode capacitative Ca(2+) channels. Immunoconfocal images showed that while Trp6 was present in the postacrosomal region and could be involved in sperm AR, expression of Trp1 and Trp3 was confined to the flagellum, suggesting that they may serve sperm to regulate important Ca(2+)-dependent events in addition to the AR. Likewise, one of these proteins (Trp1) co-immunolocalized with caveolin-1, a major component of caveolae, a subset of lipid rafts potentially important for signaling events and Ca(2+) flux. Furthermore, by using fluorescein-coupled cholera toxin B subunit, which specifically binds to the raft component ganglioside GM1, we identified caveolin- and Trp-independent lipid rafts residing in the plasma membrane of mature sperm. Notably, the distribution of GM1 changes drastically upon completion of the AR. PMID- 11734219 TI - BMP signaling regulates Nkx2-5 activity during cardiomyogenesis. AB - Nkx2-5 regulates the transcription of muscle-specific genes during cardiomyogenesis. Nkx2-5 expression can induce cardiomyogenesis in aggregated P19 cells but not in monolayer cultures. In order to investigate the mechanism by which cellular aggregation regulates Nkx2-5 function, we examined the role of bone morphogenetic protein 4 (BMP4). We showed that the expression of the BMP inhibitor, noggin, was sufficient to inhibit the induction of cardiomyogenesis by Nkx2-5 during cellular aggregation. Furthermore, soluble BMP4 could activate Nkx2 5 function in monolayer cultures, resulting in the formation of cardiomyocytes. Therefore, BMP signaling is necessary and sufficient for the regulation of Nkx2-5 activity during cardiomyogenesis in P19 cells. PMID- 11734220 TI - YFH1-mediated iron homeostasis is independent of mitochondrial respiration. AB - The human gene frataxin and its yeast homolog YFH1 affect mitochondrial function. Deficits in frataxin result in Friedreich ataxia, while deletion of YFH1 results in respiratory incompetence. We determined that as long as respiratory incompetent yeast express Yfh1p they do not accumulate excessive mitochondrial iron. Deletion of YFH1 in respiratory incompetent yeast results in mitochondrial iron accumulation, while the reintroduction of Yfh1p results in mitochondrial iron export. Further, overexpression of Yfh1p has no effect on oxygen consumption in wild-type yeast grown in either fermentative or respiratory carbon sources. We conclude that the effect of Yfh1p on mitochondrial iron metabolism is independent of respiratory activity. PMID- 11734221 TI - Molecular cloning and characterization of six novel isoforms of human Bim, a member of the proapoptotic Bcl-2 family. AB - Bim protein is one of the BH3-only proteins, members of the Bcl-2 family that have only one of the Bcl-2 homology regions, BH3. BH3-only proteins are essential initiators of apoptotic cell death. Thus far, three isoforms of Bim have been reported, i.e. Bim(EL), Bim(L) and Bim(S). Here we report the cloning and characterization of six novel isoforms of human Bim, designated as Bimalpha1, alpha2, and beta1-beta4, which are generated by alternative splicing. Unlike the three known isoforms, none of these novel isoforms contained a C-terminal hydrophobic region. Among the novel isoforms, only Bimalpha1 and alpha2 contained a BH3 domain and were proapoptotic, although less potent than the classical isoforms. These two isoforms localized, at least in part, in mitochondria when transiently expressed in HeLa cells as a green fluorescent protein-fused form. These results suggest that the BH3 domain is necessary for induction of apoptosis and mitochondrial localization but not sufficient for the full proapoptotic activity. While the classical isoforms were always predominantly expressed in transformed cells, expression profiles of bim isoforms were highly variable among normal tissues at least in humans, suggesting a tissue-specific transcriptional regulation of bim. PMID- 11734222 TI - A novel type 2C protein phosphatase from the human fungal pathogen Candida albicans. PMID- 11734223 TI - Does the western world still take human rights seriously? PMID- 11734224 TI - Examining informed consent to cancer clinical trials. PMID- 11734225 TI - Place of pegvisomant in acromegaly. PMID- 11734226 TI - Rho GTPases as early markers for tumour progression. PMID- 11734227 TI - Is breast MRI mature enough to be recommended for general use? PMID- 11734228 TI - Problems with diagnostic criteria for multiple sclerosis. PMID- 11734229 TI - Preoperative assessment: from tribalism to cooperation. PMID- 11734230 TI - Beta-cell function in new-onset type 1 diabetes and immunomodulation with a heat shock protein peptide (DiaPep277): a randomised, double-blind, phase II trial. AB - BACKGROUND: Type 1 diabetes results from autoimmune destruction of insulin producing pancreatic beta cells. The 60 kDa heat-shock protein (hsp60) is one of the known target self antigens. An immunomodulatory peptide from hsp60, p277, arrested beta-cell destruction and maintained insulin production in newly diabetic NOD mice. We did a randomised, double-blind, phase II study of peptide treatment in patients with newly diagnosed (<6 months) type 1 diabetes. METHODS: 35 patients with type 1 diabetes and basal C-peptide concentrations above 0.1 nmol/L were assigned subcutaneous injections of 1 mg p277 and 40 mg mannitol in vegetable oil (DiaPep277; n=18) at entry, 1 month, and 6 months, or three placebo injections (mannitol in vehicle; placebo; n=17). The primary endpoint was glucagon-stimulated C-peptide production. Secondary endpoints were metabolic control and T-cell autoimmunity to hsp60 and to p277 (assayed by cytokine secretion). 31 patients completed 10 months of follow-up and were included in the intention-to-treat analysis. FINDINGS: At 10 months, mean C-peptide concentrations had fallen in the placebo group (n=16) but were maintained in the DiaPep277 group (n=15; 0.26 [SD 0.11] vs 0.93 [0.35] nmol/L; p=0.039). Need for exogenous insulin was higher in the placebo than in the DiaPep277 group (0.67 [0.33] vs 0.43 [0.17] U/kg; p=0.042). Haemoglobin A1c concentrations were low (around 7%) in both groups. T-cell reactivity to hsp60 and p277 in the DiaPep277 group showed an enhanced T-helper-2 cytokine phenotype. No adverse effects were noted. INTERPRETATION: Although this study was small, treatment of newly diagnosed type 1 diabetes with DiaPep277 seems to preserve endogenous insulin production, perhaps through induction of a shift from T-helper-1 to T-helper-2 cytokines produced by the autoimmune T cells. PMID- 11734231 TI - Long-term treatment of acromegaly with pegvisomant, a growth hormone receptor antagonist. AB - BACKGROUND: Pegvisomant is a new growth hormone receptor antagonist that improves symptoms and normalises insulin-like growth factor-1 (IGF-1) in a high proportion of patients with acromegaly treated for up to 12 weeks. We assessed the effects of pegvisomant in 160 patients with acromegaly treated for an average of 425 days. METHODS: Treatment efficacy was assessed by measuring changes in tumour volume by magnetic resonance imaging, and serum growth hormone and IGF-1 concentrations in 152 patients who received pegvisomant by daily subcutaneous injection for up to 18 months. The safety analysis included 160 patients some of whom received weekly injections and are excluded from the efficacy analysis. FINDINGS: Mean serum IGF-1 concentrations fell by at least 50%: 467 mg/L (SE 24), 526 mg/L (29), and 523 mg/L (40) in patients treated for 6, 12 and 18 months, respectively (p<0.001), whereas growth hormone increased by 12.5 mg/L (2.1), 12.5 mg/L (3.0), and 14.2 mg/L (5.7) (p<0.001). Of the patients treated for 12 months or more, 87 of 90 (97%) achieved a normal serum IGF-1 concentration. In patients withdrawn from pegvisomant (n=45), serum growth hormone concentrations were 8.0 mg/L (2.5) at baseline, rose to 15.2 mg/L (2.4) on drug, and fell back within 30 days of withdrawal to 8.3 mg/L (2.7). Antibodies to growth hormone were detected in 27 (16.9%) of patients, but no tachyphylaxis was seen. Serum insulin and glucose concentrations were significantly decreased (p<0.05). Two patients experienced progressive growth of their pituitary tumours, and two other patients had increased alanine and asparate aminotransferase concentrations requiring withdrawal from treatment. Mean pituitary tumour volume in 131 patients followed for a mean of 11.46 months (0.70) decreased by 0.033 cm(3) (0.057; p=0.353). INTERPRETATION: Pegvisomant is an effective medical treatment for acromegaly. PMID- 11734232 TI - Correlation between reduction in plasma HIV-1 RNA concentration 1 week after start of antiretroviral treatment and longer-term efficacy. AB - BACKGROUND: Early assessment of antiretroviral drug efficacy is important for prevention of the emergence of drug-resistant virus and unnecessary exposure to ineffective drug regimens. Current US guidelines for changing therapy are based on measurements of plasma HIV-1 RNA concentrations 4 or 8 weeks after the start of treatment with cut-off points of 0.75 or 1.00 log, respectively. We investigated the possibility of assessing drug efficacy from measurements of plasma HIV-1 concentrations made during the first week on therapy. METHODS: The kinetics of virus decay in plasma during the first 12 weeks of treatment was analysed for 124 HIV-1-infected patients being treated for the first time with a protease inhibitor. Patients with a continuous decline of HIV-1 concentrations and in whom HIV-1 was either undetectable or declined by more than 1.5 log at 12 weeks were defined as good responders; the rest were poor responders. FINDINGS: The individual virus decay rate constants (k) at day 6 correlated significantly (r>0.66, p<0.0001) with changes in HIV-1 concentrations at 4, 8, and 12 weeks, and correctly predicted 84% of the responses with a cut-off value of k=0.21 per day (in log scale). Reduction in plasma HIV-1 of less than 0.72 log by day 6 after initiation of therapy predicted poor long-term responses in more than 99% of patients. INTERPRETATION: These results suggest that changes in HIV-1 concentration at day 6 after treatment initiation are major correlates of longer term virological responses. They offer a very early measure of individual long term responses, suggesting that treatment could be optimised after only a few days of therapy. PMID- 11734233 TI - Relation between depression after coronary artery bypass surgery and 12-month outcome: a prospective study. AB - BACKGROUND: The association of depression with cardiac events has been investigated mainly in community cohorts, in patients undergoing catheterisation, or in patients who have had myocardial infarction. We have assessed the effect of depression on outcomes after coronary artery bypass graft (CABG) surgery. METHODS: In a prospective study, we followed up for 1 year 207 men and 102 women, who had undergone coronary artery bypass graft surgery. We assessed depression with a structured psychiatric interview (diagnostic interview schedule) and a questionnaire (Beck depression inventory) before discharge. Cardiac events included angina or heart failure that needed admission to hospital, myocardial infarction, cardiac arrest, percutaneous transluminal coronary angioplasty, repeat CABG, and cardiac mortality. Non-cardiac events consisted of all other reasons for mortality or readmission. FINDINGS: 63 patients (20%) met modified diagnostic statistical manual IV criteria for major depressive disorder. At 12 months, 17 (27%) of these patients had a cardiac event compared with 25 of 246 (10%) who were not depressed (p<0.0008). Five variables had significant univariate associations with cardiac events: sex, living alone, low ejection fraction (<0.35), length of hospital stay, and depression. In a Cox proportional hazard model with these five and two other variables of cardiac severity, major depressive disorder (risk ratio 2.3 [95% CI 1.17-4.56]), low ejection fraction (2.3 [1.07-5.03]), and female sex (2.4 [1.24-4.44]) were associated with adverse outcomes. Depression did not predict deaths or admissions for non-cardiac events. INTERPRETATION: Depression is an important independent risk factor for cardiac events after CABG surgery. PMID- 11734234 TI - Clinical picture: Ruptured intracerebral abscess. PMID- 11734235 TI - Quality of informed consent in cancer clinical trials: a cross-sectional survey. AB - BACKGROUND: Investigators have to obtain informed consent before enrolling participants in clinical trials. We wanted to measure the quality of understanding among participants in clinical trials of cancer therapies, to identify correlates of increased understanding, and to assess providers' beliefs about clinical research. We also sought evidence of therapeutic misconceptions in participants and providers. METHODS: We sent a standard questionnaire to 287 adult patients with cancer who had recently enrolled in a clinical trial at one of three affiliated institutions, and surveyed the provider who obtained each patient's consent. FINDINGS: 207 of 287 (72%) patients responded. 90% (186) of these respondents were satisfied with the informed consent process and most considered themselves to be well informed. Nevertheless, many did not recognise non-standard treatment (74%), the potential for incremental risk from participation (63%), the unproven nature of the treatment (70%), the uncertainty of benefits to self (29%), or that trials are done mainly to benefit future patients (25%). In multivariate analysis, increased knowledge was associated with college education, speaking only English at home, use of the US National Cancer Institute consent form template, not signing the consent form at initial discussion, presence of a nurse, and careful reading of the consent form. Only 28 of 61 providers (46%) recognised that the main reason for clinical trials is benefit to future patients. INTERPRETATION: Misconceptions about cancer clinical trials are frequent among trial participants, and physician/investigators might share some of these misconceptions. Efforts to educate providers and participants about the underlying goals of clinical trials are needed. PMID- 11734236 TI - Diastrophic dwarfism and pregnancy. PMID- 11734237 TI - Haematological response to parvovirus B19 infection in homozygous sickle-cell disease. AB - Infection with human parvovirus B19 is known to cause aplastic crises in patients with homozygous sickle-cell disease. We studied the haematological consequences of parvovirus B19 infection in 280 such patients who had been followed up from birth in Jamaica. Evidence of seroconversion was routinely sought with a baculovirus-based, enzyme immunoassay in serum samples taken during aplastic crises and in all stored annual serum samples. 70% of patients had seroconverted by age 20 years; of 177 infections, haematological change was typical of aplastic crises in 118 (67%), minor in 16 (9%), and not discernible in 43 (24%). This assay increased the detection of unsuspected seroconversion-an observation important in planning a strategy for parvovirus B19 immunisation. PMID- 11734238 TI - Burkholderia cepacia complex genomovars and pulmonary transplantation outcomes in patients with cystic fibrosis. AB - Burkholderia cepacia is a group of organisms that comprises seven genotypically distinct species (B cepacia genomovars I-VII), which are collectively known as the B cepacia complex. Preoperative infection with B cepacia is associated with a poor prognosis in lung transplant recipients with cystic fibrosis. Many centres do not, therefore, offer transplants to these individuals. Our aim was to ascertain whether or not post-transplant mortality is affected by pretransplant genomovar status. We studied archived isolates with PCR-based methods, and recorded excessive mortality in patients infected with B cepacia genomovar III, but not in those infected with other genomovars. PMID- 11734239 TI - Cytological regression and clearance of high-risk human papillomavirus in women with an abnormal cervical smear. AB - We studied the natural course of high-risk human papillomavirus (HPV) infection and cytological regression in women referred for colposcopy because of abnormal cervical smears. We found that high-risk HPV clearance preceded regression of cervical lesions by an average of 3 months. The cumulative 1-year rate of cytological regression was similar in women with mild and moderate dyskaryotic cervical smears. Thus, retesting of high-risk HPV after 6 months in women with mild to moderate dyskaryosis predicts cytological regression. PMID- 11734240 TI - Infliximab-induced aseptic meningitis. AB - We report an episode of aseptic meningitis in a 53-year-old man, who was treated with infliximab for active rheumatoid arthritis. He had acute, severe muscle pain after initial infusion of the drug, and similar symptoms with a transient lymphocytic meningitis after a subsequent infusion. We measured no change in antibodies to nuclei, DNA, or to neurones. Functional antibodies to infliximab were not induced and concentrations of tumour necrosis factor a in spinal fluid were not raised. This adverse reaction to infliximab might have been caused by inability of the drug to enter the central nervous system. PMID- 11734241 TI - European Parliament backs human cloning ban. PMID- 11734242 TI - UN group kickstarts worldwide ban on human cloning. PMID- 11734243 TI - pH has a role in cystic fibrosis infections. PMID- 11734244 TI - High-tech eyewear ease strain of doing surgery. PMID- 11734245 TI - European action on antimicrobial resistance. PMID- 11734247 TI - Renewed interest in leucocyte count as prognostic indicator after AMI. PMID- 11734248 TI - US government undercuts Oregon's assisted-suicide law. PMID- 11734250 TI - Panel rejects u-turn on gay blood donor ban. PMID- 11734249 TI - Blood, politics, and the American Red Cross. PMID- 11734251 TI - EU ministers set stricter rules on donor blood. PMID- 11734252 TI - UK league table reveals variations in mortality rate after heart surgery. PMID- 11734253 TI - US university finds scientist flouted rules for clinical trials. PMID- 11734256 TI - UN pleads for support for aid workers in Afghanistan after Taliban collapse. PMID- 11734257 TI - China responds to increasing HIV/AIDS burden and holds landmark meeting. PMID- 11734258 TI - Gonadotropin-releasing-hormone-receptor antagonists. AB - Pulsatile gonadotropin-releasing hormone (GnRH) stimulates the pituitary secretion of both luteinising hormone (LH) and follicle-stimulating hormone (FSH) and thus controls the hormonal and reproductive function of the gonads. Blockade of GnRH effects may be wanted for a variety of reasons-eg, to prevent untimely luteinisation during assisted reproduction or in the treatment of sex-hormone dependent disorders. Selective blockade of LH/FSH secretion and subsequent chemical castration have previously been achieved by desensitising the pituitary to continuously administered GnRH or by giving long-acting GnRH agonists. Only recently have GnRH-receptor antagonists, that immediately block GnRH's effects, been developed for clinical use with acceptable pharmacokinetic, safety, and commercial profiles. In assisted reproduction, these compounds seem to be as effective as established therapy but with shorter treatment times, less use of gonadotropic hormones, improved patient acceptance, and fewer follicles and oocytes. All current indications for GnRH-agonist desensitisation may prove to be indications for a GnRH antagonist, including endometriosis, leiomyoma, and breast cancer in women, benign prostatic hypertrophy and prostatic carcinoma in men, and central precocious puberty in children. However, the best clinical evidence so far has been in assisted reproduction and prostate cancer. PMID- 11734259 TI - Biotechnology and new companies arising from academia. AB - 20 years ago, an academic biomedical scientist or clinician who set up a company would probably have been perceived by colleagues as "on the make" and rather unacademic-"not one of us", in other words. Nowadays, academics who have started companies are commonplace, and in some universities the businessman-academic is becoming the norm, although still far more common in the USA than in Europe. At best, the opportunity to capitalise on a discovery has the potential to motivate research workers, provide greater funding for research, and ultimately create wealth. At worst, the spawning of a company from within academia has the potential to use public employees, space, and equipment for personal gain, and divert academics from the pursuit of profound scientific questions into more immediate product-driven research or even marketing dressed up as research. Here, I discuss some of the issues surrounding biotechnology and spin-off companies originating in academia. PMID- 11734261 TI - Doctors in the UK: their new professionalism and its regulatory framework. AB - The new professionalism evolving in UK medicine is fundamentally different from the past. Founded on the General Medical Council's (GMC) Good Medical Practice, the new professionalism is an explicit statement of professional duties, responsibilities, values, and standards for doctors, developed and agreed on by the public and the profession. Compliance is being secured by embedding the culture of Good Medical Practice into medical education and by linking it directly with medical registration (licensure). From 2004 doctors will have their abilities assessed regularly by peers and by members of the public to ensure that they remain up-to-date and fit to practise medicine. Disciplinary action will follow if they are not. To ensure patient safety, the new professionalism requires professional leadership with great public input to medical regulation, a modern GMC, and a closer fit between the licensure of doctors and the quality assurance of the organisations in which they work. PMID- 11734262 TI - Effects of fenofibrate and gemfibrozil on plasma homocysteine. PMID- 11734266 TI - Hypopituitarism after head injury. PMID- 11734263 TI - Effects of fenofibrate and gemfibrozil on plasma homocysteine. PMID- 11734267 TI - Familial malignant mesothelioma. PMID- 11734269 TI - D-lactate encephalopathy. PMID- 11734270 TI - Sentinel-node biopsy in breast cancer. PMID- 11734271 TI - Sentinel node biopsy in breast cancer. PMID- 11734273 TI - Sentinel node biopsy in breast cancer. PMID- 11734274 TI - Sentinel node biopsy in breast cancer. PMID- 11734275 TI - Delayed sexual development in adolescents. PMID- 11734276 TI - Improved management of asthma. PMID- 11734278 TI - Art and science of clinical knowledge. PMID- 11734279 TI - Declaration of Commitment against HIV/AIDS. PMID- 11734280 TI - Declaration of Commitment against HIV/AIDS. PMID- 11734281 TI - Prevention of antiretroviral anarchy in sub-Saharan Africa. PMID- 11734282 TI - Computerised study applications. PMID- 11734283 TI - Abuse of people trying to protect children from abuse. PMID- 11734285 TI - Prevention of antiretroviral anarchy in sub-Saharan Africa. PMID- 11734286 TI - Effect of cataract on amblyopic eye. PMID- 11734296 TI - Sandbank. PMID- 11734297 TI - Establishment of novel B-cell precursor leukemia sister cell lines NALM-36 and NALM-37: shift of immunoglobulin phenotype to double light chain positive B-cell. AB - Two novel B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) sister cell lines, designated NALM-36 and NALM-37, were established from the peripheral blood (at diagnosis) and bone marrow (at relapse) of a 37-year-old woman with ALL. Immunophenotyping showed BCP type III pre-B cell characteristics including TdT, CD10, CD19, CD22, CD79a and HLA class II. T cell and myeloid-associated antigens tested were negative except CD5 which was 100% positive for both cell lines. The surrogate light chains lambda5 and VpreB were positive for both cell lines. Cytogenetic analysis of NALM-36 revealed an abnormal karyotype with 46, XX, add(1)(q?42), -14, +mar. Southern blot analysis of the immunoglobulin (Ig) genes status of NALM-36 at 10 months after establishment showed germ line configuration of the kappa light chain gene, and rearrangement of the lambda light and mu heavy chain genes. At 16 months we detected a phenotypic shift of Ig chain protein expression from a BCP-III pre-B cell phenotype to a BCP-IV mature B cell phenotype, with kappa and lambda double Ig light chain and mu heavy chain expression, both on the cell surface and in the cytoplasm. We designated this subline as NALM-36KL. Authenticity of the NALM-36KL, NALM-36 and NALM-37 cell lines was demonstrated by DNA fingerprinting. The extensive characterization of the sister cell lines suggests that these three novel cell lines, derived from a single patient, may represent unique and relevant in vitro model systems for BCP type leukemia cells. They may provide useful models and unprecedented opportunities for analyzing the multitude of biological aspects of normal and neoplastic B-lymphocytes and their precursors. PMID- 11734298 TI - Shift of immunoglobulin phenotypes in B-cell precursor cell lines. AB - Rearrangement of immunoglobulin chain genes is a hallmark of mammalian B cell development and differential expression of immunoglobulin chains has aided in the diagnosis and prognostication of patients with ALL. Matsu et al. describe two novel precursor B-cell lines with shift of expression of immunoglobulin phenotypes at diagnosis and relapse. The impact of this observation on diagnosis and the elucidation of the biology of the neoplastic cells will be discussed. PMID- 11734299 TI - Evaluation of engraftment by ABO genotypic analysis of erythroid burst-forming units after bone marrow transplantation. AB - Six patients received an allogeneic bone marrow transplant from HLA-identical ABO mismatched donors. ABO genotype of erythroid burst-forming units (BFU-E) from peripheral blood was analyzed using polymerase chain reaction with sequence specific primers (PCR-SSP). After bone marrow transplantation (BMT), engraftment of donor cells by ABO genotypic analysis of BFU-E was compared with ABO phenotypic analysis of red blood cells (RBCs). During the early stage after BMT, ABO genotype of BFU-E in the recipients converted to that of the donors. In contrast, mixed ABO phenotype of RBCs persisted for about 3 months. In one patient, autologous hemopoietic cell recovery was detected by the ABO genotypic analysis before clinical manifestation. ABO genotypic analysis of BFU-E is relevant for enagraftment after ABO-mismatched BMT. PMID- 11734300 TI - Bone and bone-marrow interactions: haematological activity of osteoblastic growth peptide (OGP)-derived carboxy-terminal pentapeptide. Mobilizing properties on white blood cells and peripheral blood stem cells in mice. AB - Osteogenic growth peptide (OGP) increases blood and bone marrow cellularity in mice, and enhances engraftment of bone marrow transplant. Carboxy-terminal pentapeptide (OGP10-14) holds several properties of full-length polypeptide. We evaluated whether synthetic OGP-derived pentapeptide (sOGP10-14) has some activity on peripheral blood cell recovery after cyclophosphamide-induced aplasia, and on stem cell mobilization. Peripheral blood stem cell (PBSC) mobilization was evaluated by administering granulocyte-colony stimulating factor (G-CSF) or sOGP10-14 after cyclophosphamide (CTX) injection. Haematological parameters and CD34/Sca-1 positive cells were sequentially evaluated. Colony forming tests were performed in bone marrow cells from CTX-, G-CSF- and sOGP10-14 treated mice. sOGP10-14 was able to enhance band cells and monocyte recovery after cyclophosphamide administration. White blood cell (WBC) counts reached the maximum peak by day +10 but, on day +7, a significant recovery was already detected in sOGP10-14 treated mice. On day +10 the WBC increase in sOGP10-14 treated mice was comparable to that found in G-CSF treated ones. Moreover, CD34/Sca-1 positive early precursors were significantly mobilized by sOGP10-14 compared to the control group. In sOGP10-14-treated mice, the colony-forming unit granulocyte-macrophage-megakaryocyte (GEMM-CFU) and burst-forming unit-erythroid (BFU-E) were significantly increased in bone marrow cells in comparison to mice treated with CTX only. These results suggest a central role of sOGP10-14 in bone and bone marrow interaction, and a possible role of sOGP10-14 as a mobilizing agent. PMID- 11734301 TI - Augmented expression of P-gp/multi-drug resistance gene by all-trans retinoic acid in monocytic leukemic cells. AB - P-glycoprotein (P-gp)/multi-drug resistance 1 (MDR1) gene is recognized to be, at least in part, responsible for the refractoriness to chemotherapy of leukemia. The transcriptional mechanism of MDR1 gene is largely unknown. However, recent reports have clarified that early growth response 1 gene (Egr1) positively regulates MDR1 transcription, while Wilms' tumor suppressor gene (WT1) does negative regulation of MDR1 gene expression in 12-O-tetradecanoylphorbol-13 acetate treated K562 cells. In addition, Egr1 and WT1 are structurally related transcription factors and bind to quite similar DNA sequences. Our study of mRNA expression profile of Egr1, WT1 and MDR1 in fresh AML samples demonstrated that there are disease-specific patterns. Egr1 mRNA was frequently and strongly expressed in monocytic leukemia cells, especially in AML M4 cells. WT1 mRNA was undetectable in t(8;21) AML cells. mRNA expression of MDR1 was frequent in AML M1 and t(8;21) AML cells, in which the expression level was highest in AML M1 and was low in monocytic leukemia (M4 and M5). Then, expression level of MDR1 was inversely correlated with Egr1. By liquid culture of leukemia cell lines and fresh AML cells with the addition of all-trans retinoic acid (ATRA), modulation of P-gp/MDR1 and Egr1 was observed and the pattern of modulation was divided into four groups: (1) blastic AML type, in which distinct expression of P-gp/MDR1 and CD34 was not influenced by ATRA; (2) t(8;21)AML type, in which P-gp/MDR1 expression was augmented by ATRA, while CD34 was kept high; (3) AML M3 type, in which P-gp/MDR1 expression was reduced with granulocytic differentiation by ATRA; (4) monocytic AML type, in which P-gp/MDR1 expression was augmented by ATRA, while CD34 expression decreased, and strong Egr1 expression was downregulated just prior to the augmentation of P-gp/MDR1 expression. WT1 expression was not influenced by the addition of ATRA in each group. Previous reports have suggested that P-gp/MDR1 plays an important role in resistance to chemotherapy, and is recognized as one of the stem cell marker. However, P-gp/MDR1 expression augmented by ATRA, which was observed in monocytic AML, was recognized as a functional molecule of mature monocyte/macrophage, because CD34 expression decreased and CD13 expression increased by ATRA. Finally, expression of P-gp/MDR1 in monocytic leukemia, which was functionally confirmed by Rh123 efflux study, was thought to be closely related to the characteristic modulation of Egr1 expression by ATRA. PMID- 11734302 TI - In vitro testing of the leukaemia monoclonal antibody WM-53 labeled with alpha and beta emitting radioisotopes. AB - We report the preparation and testing of a new alpha emitting radio immunoconjugate (RIC) against acute myeloid leukaemia (AML) using CD33 positive monoclonal antibody WM-53 (specific for HL-60 cell line). Using cyclic anhydride of diethylenetriaminepentacetic acid (cDTPAa) as chelator, antibody was labeled with 213Bi (alpha), 149Tb (alpha), 153Sm (beta) and 152Tb (positron). In vitro testing showed high labeling efficiency (90-95%) and stability (11-19% leaching) with immunoreactivity virtually the same before and after labeling. DNA synthesis data and MTS cell survival were compared for all RICs. Only the alpha emitter was found to be capable of inhibiting DNA synthesis and had selective cell kill with activity as low as 2-3 microCi. The high stability and outstanding cytotoxicity of the 213Bi conjugate provides the basis for targeted alpha therapy for the control of metastatic and disseminated cancer such as AML. PMID- 11734303 TI - Monoclonal antibody to human chronic myeloid leukemia cell line MOLM-7 specifically reacts with an antigen of apoptotic cells. AB - Monoclonal antibody 2E12 was prepared by immunization of mice with fresh cells of chronic myeloid leukemia cell line MOLM-7. A panel of 15 leukemic cell lines (myeloid, promyelocytic, erythroid, B and T lymphoid) and numerous cultured patient's leukemia and myeloma cells were tested for reactivity with 2E12 antibody. A subset of cells in all cell lines and various number of patient's cells cultivated for 10 days or more were 2E12 positive. KG-1 and HL-60 cell lines were treated by camptothecin (CAM) (5 microg/ml, 4 h), washed and further cultivated without CAM. After 24 and 48 h in culture a considerable increase of 2E12 positivity was detected both in KG-1 and HL-60 cells, which well correlated with the increase of APO2.7 positivity and the sub-G1 peaks. The 2E12 positive cells were morphologically the same as cells in PCD, possibly apoptosis. We suggest that the 2E12 antibody detects a strong antigen on apoptotic cells which could be a part of the signaling process for ingestion by phagocytes. PMID- 11734304 TI - c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) is required for mitoxantrone- and anisomycin-induced apoptosis in HL-60 cells. AB - Programmed cell death, or apoptosis, has emerged as a common mechanism by which cells respond to chemotherapeutic drugs. However, the signaling mechanisms that mediate drug-induced apoptosis are still widely unknown. Mitogen-activated protein kinase (MAPK) signaling cascades trigger stimulus-specific responses in cells with ERK being associated with proliferation and differentiation, and JNK/SAPK and p38 mediating stress and apoptotic responses. Here, we found that mitoxantrone and anisomycin stimulated a dose- and time-dependent induction of JNK/SAPK activity, and to a lesser extent p38 activity, that preceded the appearance of apoptosis as measured by internucleosomal DNA fragmentation. These compounds did not induce ERK activity. We further demonstrated that p38 activity was not involved in the induction of apoptosis since the use of the p38 inhibitor, SB203580, did not prevent drug-induced apoptotic DNA fragmentation. Additionally, direct inhibition of JNK/SAPK signaling through the use of dominant negative MKK4/SEK1 (SEK-AL) inhibited mitoxantrone- and anisomycin-induced apoptosis. These results suggest that mitoxantrone- and anisomycin-induced apoptosis is dependent on JNK/SAPK, but not p38, activity. PMID- 11734305 TI - Protein kinase C isoforms in normal and leukemic neutrophils: altered levels in leukemic neutrophils and changes during myeloid maturation in chronic myeloid leukemia. AB - Protein kinase C (PKC) is reported to play a role in maturation of the myeloid cell and functions of the mature neutrophil. The neutrophils in chronic myeloid leukemia (CML) exhibit defects in several functions. As a step towards understanding the role of PKC in the defects in function of the leukemic cells, this study investigates the expression of PKC isoforms, their subcellular distribution, levels and kinase activity in the normal and leukemic neutrophils. It also investigates changes in representative PKC isoforms during myeloid maturation. This study confirms the presence of PKC alpha, beta and delta and shows, for the first time, the presence of non conventional PKC isoform theta, atypical PKC isoform lambda/iota and PKC isoform mu in normal human neutrophils. In unstimulated cells all the detected PKC isoforms showed a predominantly cytosolic localisation in normal and CML neutrophils. Cytosol-membrane distribution of PKC alpha and delta were significantly altered in leukemic neutrophils as compared to normal cells. Cytosolic levels of all PKC isoforms were reduced in CML neutrophils with PKC alpha, beta, iota, theta, and mu showing a significant decrease. Cytosolic levels of PKC delta contrary to the trend observed for other PKC isoforms showed a slight increase in CML cells, while its membrane levels were significantly reduced in CML neutrophils. Total PKC kinase activity in CML neutrophil cytosol was significantly reduced, while specific kinase activity of two representative isoforms, PKC alpha and delta, from normal and CML neutrophils were similar, thereby increasing the significance of the altered levels of PKC isoforms in CML, and highlighting their role in the defects in function exhibited by the leukemic neutrophils. The levels of PKC delta and iota increased and decreased respectively as the leukemic myeloid cell matured from the blast to the neutrophil, while the levels of PKC alpha and beta were not altered. This suggests a role for PKC delta and iota in the maturation of the leukemic myeloid cell. PMID- 11734306 TI - Immunohistochemical studies of c-kit, transforming growth factor-beta, and basic fibroblast growth factor in mast cell disease. AB - In 58 patients with mast cell disease (MCD) and three with basophilic leukemia, bone marrow (54 cases) or skin tissue (four cases) was studied immunohistochemically for expression of Kit (c-kit protein), the different isomers of transforming growth factor-beta (TGF-beta), basic fibroblast growth factor (bFGF), and their respective receptors. Kit was expressed in all cases of MCD but in none of basophilic leukemia. Expression pattern of cytokines and their receptors was variable in systemic MCD with (SMCD-HD) or without (SMCD) associated hematologic disorder. However, type I TGF-beta receptor (TGFbeta1R) was not expressed in 30% of SMCD-HD patients or in patients with mast cell leukemia, but the remaining cases of MCD showed near uniform expression. The associated hematologic disorders in TGFbeta1R-negative cases of SMCD-HD were prognostically less favorable than those associated with TGFbeta1R-positive cases of SMCD-HD. The results confirm the diagnostic value of Kit immunohistochemistry in MCD and suggest a biologically relevant heterogeneity in TGFbeta1R expression among patients with SMCD-HD. PMID- 11734307 TI - PK11195, a peripheral benzodiazepine receptor ligand, chemosensitizes acute myeloid leukemia cells to relevant therapeutic agents by more than one mechanism. AB - Like Bcl-2, peripheral benzodiazepine receptors (pBzRs) reside in mitochondrial pores, are frequently over-expressed in tumor cells, and can protect cells from apoptotic cell death. We now show that the high-affinity, pBzR-specific ligand, PK11195, chemosensitizes AML cells to relevant chemotherapeutics, but is relatively non-toxic as a single agent, and does not chemosensitize normal myeloid cells. PK11195 can block p-glycoprotein efflux in AMLs, contributing to increased daunomycin toxicity in efflux-competent AMLs, but can also sensitize AMLs to cytarabine and DNR-sensitize efflux-incompetent AMLs, presumably via mitochondrial pore effects documented in other models. Therefore, PK11195 might contribute to improved clinical outcomes in AML. PMID- 11734308 TI - Overcoming drug resistance: targeting more than one site. PMID- 11734309 TI - Rapid clonal development in a relapsed CML 11 years post replete allogeneic bone marrow transplantation. AB - Long-term survival of chronic myeloid leukaemia (CML) patients transplanted in chronic phase with a replete marrow have been described previously. The same success has not been achieved with patients in more advanced stages of the disease. We describe a CML patient who received an allogeneic bone marrow transplantation in accelerated phase. Cytogenetic and molecular analysis confirmed donor chimaerism, and the absence of the BCR-ABL mRNA. Almost 12 years post-transplant relapse was noted. Cytogenetic analyses showed a complex evolving karyotype. These findings are correlated with the longitudinal molecular analysis utilising real-time and VNTR PCR. PMID- 11734310 TI - Primary lymphoma of the heart. A case report and review of the literature. AB - Primary cardiac lymphoma (PCL) is a rare and usually fatal neoplasm, which may cause syncope, arrhythmia, heart failure and pericardial effusion as presenting clinical complaints. A case of PCL in a 72-year-old man with moderate aortic stenosis is presented. The patient was investigated because of pericardial effusion and diagnosis of diffuse large B-cell lymphoma was obtained by open chest biopsy of the heart. Fatal ventricular arrhythmia developed the day after the first course of chemotherapy. Clinical presentations and diagnostic approach of this rare tumour are discussed. While chemotherapy is the only effective treatment of PCL, early post-chemotherapy phase should be considered critical in patients with PCL, as suggested by other reported fatal complications in this period. PMID- 11734311 TI - A review of the cytogenetics of 58 pediatric brain tumors. AB - We describe the cytogenetic results of 58 pediatric central nervous system (CNS) tumors of variable histology, investigated between 1992 and 2000. Successful cytogenetics were obtained for 53 patients, with clonal chromosome abnormalities demonstrated in 25. Notable findings included (1) 2p abnormality in four primitive neuroectodermal tumors (PNET); (2) 1p loss in four low-grade gliomas and two PNET; (3) telomeric associations in one pilocytic astrocytoma; (4) chromosome 7 gain in four astrocytomas and two PNET; (5) 17p loss in four PNET; (6) double minutes in one PNET and three glioblastomas; and (7) chromosome 10 loss in four PNET. Higher grade tumors demonstrated greater karyotype complexity. Low-grade tumors showed either minimal simple chromosome changes or a normal karyotype. Chromosome abnormalities were more frequent in supratentorial tumors than their infratentorial counterparts. Our results add weight to the limited existing body of cytogenetic documentation for pediatric CNS tumors and provide further evidence that 2p loss is a consistent region of chromosome involvement in PNET. We advocate further studies of CNS tumors, in particular, to evaluate the importance of 2p changes and to compare cytogenetic results for supratentorial tumors and their infratentorial counterparts. PMID- 11734312 TI - A well-differentiated liposarcoma with a new type of chromosome 12-derived markers. AB - Well-differentiated liposarcomas (WDLPS) are cytogenetically characterized by the presence of supernumerary ring or giant rod marker chromosomes. These supernumerary chromosomes are composed of amplified sequences from chromosome 12 (12q14 approximately 15) in association with amplified segments from various other chromosomes, and contain alterations of the alpha satellite sequences. We report a case of WDLPS of the lipoma-like and sclerosing subtype that contains a novel type of supernumerary marker chromosome. Instead of rings or giant rods, these cells had three apparently identical copies of a subtelocentric supernumerary marker with a size and shape similar to C-group chromosomes. Fluorescence in situ hybridization analysis revealed that the markers were composed of amplified material from 12q14 approximately 15, including the genes MDM2 and CDK4. Similar to the rings and giant rods observed in other WDLPS cases, these unusual markers had no alpha satellite repeats at the primary constriction site, but centromeric activity could be demonstrated by using anti-centromere protein C antibodies. These findings show that the supernumerary markers of WDLPS may be variable in size and shape, but consistently share the same genomic structure, specifically 12q amplified sequences together with centromere alterations, and underline the importance of molecular methods in the diagnosis of adipose tissue tumors. PMID- 11734313 TI - Analysis of the distribution and frequency of trisomy 7 in vivo in synovia from patients with osteoarthritis and pigmented villonodular synovitis. AB - Osteoarthritis (OA) and pigmented villonodular synovitis (PVNS) are disorders associated with trisomy 7. The aim of the present study was to determine the frequency and distribution of the cells with +7 in vivo by analyzing sections of paraffin-embedded synovia from patients affected by OA, PVNS, other forms of synovitis [hemorragic synovitis (HS) and chronic synovitis (CS)], and from individuals without joint disease. Fluorescence in situ hybridization (FISH), using a centromeric probe for chromosome 7, showed that the mean frequency of trisomic nuclei in 5-microm sections was highest in PVNS (9.0%), followed by CS (5.9%), OA (5.6%), and HS (4.6%), whereas trisomic nuclei were rare (0.7%) in normal tissue. When 8-microm sections were studied, the frequencies of trisomic cells in OA and control synovia increased to 6.7% and 1.5%, respectively. Trisomic nuclei were found in all cases, including those for which cytogenetic analysis of short-term cultures had not disclosed any trisomic cells. Overall, the trisomic cells were scattered within the tissue. However, small clusters of cells with +7 were found in three cases. By hematoxylin-eosin staining of the slides used for FISH analysis it could be shown that the clustered trisomic cells were proliferating synoviocytes within villous extensions of the synovial membrane. PMID- 11734314 TI - Complete cytogenetic characterization of the human breast cancer cell line MA11 combining G-banding, comparative genomic hybridization, multicolor fluorescence in situ hybridization, RxFISH, and chromosome-specific painting. AB - The MA11 cell line was established from malignant cells isolated from the bone marrow of a breast cancer patient. It metastasizes selectively to the brain in athymic mice. Since the genomic rearrangements of only a few breast cancer cell lines have been characterized completely, we analyzed MA11 cytogenetically. Because the G-banding analysis revealed a very complex karyotype with several markers and chromosomes with additional material of unknown origin, we used multicolor fluorescence in situ hybridization (M-FISH), cross-species color banding (RxFISH), comparative genomic hybridization (CGH), and chromosome specific probes to better characterize the chromosome abnormalities. The use of these FISH-based screening techniques allowed us to detect previously unsuspected chromosomal changes and determine the identity of chromosomal markers. Multicolor FISH was especially useful to identify the rearranged chromosomes, whereas RxFISH, G-banding, and CGH were instrumental in determining breakpoint positions, although some uncertainties were removed only after hybridization with chromosome specific probes. The combined analysis revealed a near-triploid karyotype with no less than 20 chromosomes demonstrating structural rearrangements. The resulting imbalances included several of those known to be common in primary breast carcinomas (gain of 1q, 8q, and 20q and loss of 8p, 11q, and 13q), indicating that the MA11 cell line may serve as a good model to study breast carcinogenesis. The full cytogenetic characterization we present may guide future searches for the mechanism of organ-selective metastasis in this model system and, possibly, also in vivo. PMID- 11734315 TI - Cytogenetic-clinicopathologic correlations in rhabdomyosarcoma: a report of five cases. AB - Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children younger than the age of 15 years. Histologically, RMS can be subdivided into two major subtypes; embryonal (E-RMS) and alveolar (A-RMS) rhabdomyosarcoma, with E-RMS being the more common. Although cytogenetic and molecular genetic findings have been reported extensively for RMS, clinicopathologic-genetic correlations among these tumors have not been reported in detail. In this report, we correlate the cytogenetic findings, including fluorescence in situ hybridization and spectral karyotyping, with pathologic findings and outcome for five RMS, including two A RMS, one E-RMS, one botryoid RMS, and one anaplastic nonclassified RMS (N-RMS). The findings in A-RMS and E-RMS generally were consistent with previous reports; however, gain of chromosome 7 in A-RMS and gain of chromosome 9 segments in E-RMS observed here have seldom been reported in the literature. Importantly, the botryoid RMS had a cytogenetic profile similar to other types of E-RMS. An add(11)(q21) observed in this tumor, together with a t(8;11)(q12 approximately 13;q21) reported previously, indicates that 11q21 rearrangements may be nonrandomly related to botryoid RMS. In addition, the N-RMS expressed a cytogenetic pattern similar to that observed in E-RMS, thus providing genetic evidence that anaplastic N-RMS is a variant of E-RMS. Finally, these cases provide cogent evidence for the diagnostic and prognostic significance of the pathologic-genetic classification of RMS. PMID- 11734316 TI - BCR/ABL fusion gene detected on 9q34 by fluorescence in situ hybridization in an acute leukemia with two BCR/ABL positive clones, one Ph-negative and one Ph positive. AB - We report cytogenetic, fluorescence in situ hybridization (FISH), and molecular analyses in the first reported case of an acute leukemia with two BCR-positive clones: one cell Ph-positive and all others Ph-negative. A BCR/ABL fusion gene on 9q34 was detected only with a BCR/ABL dual color translocation probe. These FISH interphase signals must be confirmed on a metaphase to avoid an erroneous interpretation. This observation appears to indicate a 2-step mechanism for this aberrant fusion gene localization: first, a classical t(9;22), and then the transfer of the fusion gene formed on chromosome 22 to chromosome 9 by a second translocation between the long arms of the derivative chromosomes 9q+ and 22q-, masking the first chromosome exchange. PMID- 11734317 TI - Pseudodicentric (16;12)(q11;p11.2) in a type AB (mixed) thymoma. AB - Genetic alterations of thymomas are rarely described in the literature. In this study, a previously unreported instance of aberrant karyotypic change consisting of 45,XX,pseu dic(16;12) (q11;p11.2) [cp23]/87-90,idemx2[cp4] in a Masaoka Stage II mixed thymoma or type AB thymoma affecting a 56-year-old Chinese woman is detailed. Abnormalities involving 12p containing important tumor suppressor-like genes have been documented especially in hematological malignancies. Recently, recurrent losses involving 16q, a locus known to harbor several tumor suppressor genes, have been described in type C thymomas (squamous cell carcinoma), suggesting a possible relationship between type AB thymoma and type C thymoma. Whether these genes are involved in the pathogenesis of type AB thymoma remain to be clarified and it is currently unclear if cytogenetic studies may eventually play a role in the classification of thymic tumors. PMID- 11734318 TI - Frequent gain of copy number on the long arm of chromosome 3 in human cervical adenocarcinoma. AB - We analyzed genomic aberrations in 20 cervical adenocarcinomas by comparative genomic hybridization (CGH). Most tissue samples (85%) showed DNA copy number changes; gains were more common than losses. The most consistent region of chromosomal gain was mapped to chromosome arm 3q, found in 70% of the cases, with a minimal common region of 3q28-ter. Other recurrent amplifications of genetic material were detected on 17q (45%), 1p (30%), 1q (25%), and 11q (20%). High level copy number increases were found in chromosomal regions 3q27-ter and 9pter 13. DNA losses were seldom observed, occurring primarily in underrepresented regions of chromosome arms 4q, 13q, and 18q. The presence of high-risk human papilloma virus genomes in the cervical adenocarcinoma samples was detected in 90% of the cases. However, there was no correlation between human papilloma virus type and the pattern of genomic changes. This study is the first report of CGH analysis in human cervical adenocarcinoma. Among the major genomic alterations, our results demonstrate the importance of DNA copy increases of chromosome arm 3q in the development of cervical adenocarcinoma and identify other amplified chromosomal regions that are also associated with cervical carcinogenesis. PMID- 11734319 TI - Allelic gains and losses in distinct regions of chromosome 6 in gastric carcinoma. AB - In gastric cancer, alterations in the long arm of chromosome 6 are a frequent event. Two regions of heterozygous loss have been described: 6q16.3-6q23 and 6q26 6q27. We have evaluated by microsatellite and FISH analyses the 6q status of three cell lines that we established from primary gastric carcinomas xenografted in nude mice, in order to elucidate the underlying mechanisms of 6q alterations. Alterations of the long arm of chromosome 6 were found in all three xenografts and corresponding cell lines. Allelic imbalance (AI) was found in the three cases, by microsatellite analysis. Fluorescence in situ hybridization analyses clearly demonstrated a duplication of the larger part of the 6q arm in all three cell lines. Two of the cell lines and the corresponding xenografts showed, in addition, complete loss of one of the parental alleles at the terminal part of 6q. Thus, the AI observed along the long arm of chromosome 6 is represented by gain of alleles in one distinct chromosomal segment and loss of alleles in another distinct segment. PMID- 11734320 TI - Analysis of chromosomal aberrations in large hepatocellular carcinomas by comparative genomic hybridization. AB - Hepatocellular carcinoma (HCC) is a very common and highly malignant tumor, associated mainly with chronic viral hepatitis, cirrhosis of any cause, aflatoxin exposure and ethanol consumption. The aim of this study was to map genomic aberrations in HCC by a recently developed technique: comparative genomic hybridization (CGH). We applied CGH on 17 liver specimens, of which seven were HCCs. The rest were benign liver tumors, cirrhotic and normal livers, and other liver malignancies. Our study included mainly large tumors (mean size 10.5 cm) unrelated to viral hepatitis or cirrhosis. Our CGH analysis detected genomic imbalances in 42% of HCCs. The common aberrations included DNA gains of 1q, 9p, and 8q and DNA losses of 17p, 13q, 9q, 4q, and 11q. Also, we detected trisomies 8, 9, 18 and 21, which have not been reported previously. Gains and losses of DNA found in this study probably involve oncogenes and tumor suppressor genes that play a role in the puzzle of hepatocarcinogenesis. This study also suggests a possible link between the size of the tumor and the burden of genetic changes. PMID- 11734321 TI - Deletion of 16q11 is a recurrent cytogenetic aberration in acute myeloblastic leukemia during disease progression. AB - Abnormalities of chromosome 16 other than inv(16)(p13q22), t(16;16)(p13;q22), and del(16)(q22) have not been fully characterized in acute myeloblastic leukemia (AML) and myelodysplastic syndrome (MDS). We report here the first case of AML with del(16)(q11) as a sole abnormality. A 53-year-old woman was initially diagnosed as MDS, refractory anemia with excess of blasts in transformation with normal karyotype. After sixteen months, the disease progressed to overt AML-M1. Myeloblasts were positive for CD13, CD33, and CD34, but negative for HLA-DR. Chromosome analyses of the bone marrow cells showed 46,XX,del(16)(q11) in all metaphase spreads. Multicolor spectral karyotyping also confirmed that del(16)(q11) was not derived from a cryptic translocation, but a simple deletion. Our results, together with three previously reported cases, suggest that del(16)(q11) may be one of the recurrent aberrations in AML and that it could be associated with clonal evolution or disease progression. PMID- 11734322 TI - Is 4q13 a recurring breakpoint in solitary fibrous tumors? AB - Solitary fibrous tumor (SFT) is a mesenchymal neoplasm found predominantly in the subpleural region but also in many other body sites. We report a malignant solitary fibrous tumor of the peritoneum with a 47,XY,t(4;9)(q13;p23),+5 karyotype. The chromosome 4q13 breakpoint in the presented and previously published case of pleural solitary fibrous tumor with a 46,XY,t(4;15)(q13;q26) karyotype was further characterized by fluorescence in situ hybridization analysis and localized within the 5-cM interval that was flanked by regions specific to YAC clones 761A7 and 886C11. Chromosome translocations involving chromosome 4q13 may characterize a separate cytogenetic subgroup of SFT. PMID- 11734323 TI - Chromosomal changes in locally recurrent, hormone-refractory prostate carcinomas by karyotyping and comparative genomic hybridization. AB - The genetic mechanisms of prostate cancer recurrence during hormonal therapy are largely unknown. So far, data from conventional karyotype analysis on hormone refractory prostate carcinomas have not been published, mainly because of the difficulties in obtaining fresh hormone-refractory prostate carcinoma samples and getting metaphases from them. Here, we have studied chromosomal changes in 12 locally recurrent, hormone-refractory prostate carcinomas using karyotyping and CGH that revealed genetic aberrations in all tumors. Loss of the Y chromosome was the most common (89%) finding, and tetraploidy or near-tetraploidy was detected in all tumors. Also non-random translocations were found in 56% of the tumors. The present study indicates that clonal chromosomal aberrations in hormone refractory prostate carcinomas are more common than in untreated primary tumors, and also, further studies on the frequency and significance of translocations in prostate carcinoma progression during hormonal therapy are warranted. PMID- 11734324 TI - Deletion 5p11 accompanied by multiple numerical changes in testicular lymphoma. AB - We describe a case of testicular B cell lymphoma with deletion of chromosome 5, del(5)(p11), as a sole structural abnormality. Histopathological diagnosis of the tumor was a high-grade lymphoma of the diffuse type containing cells positive for B cell specific antigen (CD20) and negative for the leukocyte common antigen (CD45). Deletion 5p may define the region of a tumor suppressor gene that could be associated with tumor progression and invasiveness and may serve as an indicator of poor prognosis in testicular lymphomas. PMID- 11734325 TI - Isochromosome (17)(q10) and translocation (4;12)(q12;p13) in a child with acute myeloid leukemia. AB - Isochromosome 17q is a commonly observed cytogenetic aberration in hematologic malignancies. Isolated isochromosome 17q usually presents as a marker of a chronic myeloid disorder, with a high propensity for transformation into acute nonlymphoblastic leukemia (ANLL). t(4;12)(q11-12;p13) is a recently described translocation, associated with ANLL, predominantly in adults. In this article, we present a case of acute myeloblastic leukemia (AML) in a 14-year-old female in which i(17q) and t(4;12)(q12;p13) were found in the leukemic clone at diagnosis. We briefly review the literature and hypothesize as to the significance of the coexistence of these cytogenetic changes. PMID- 11734326 TI - Cytogenetic biclonality in polycythemia vera: unusual and unrelated clones. PMID- 11734327 TI - Del(X)(q23) is associated with dyserythropoiesis in elderly females. PMID- 11734328 TI - Novel translocation (5;18)(q31;q23) in biphenotypic acute leukemia. PMID- 11734330 TI - BRCA2: a genetic risk factor for breast cancer. AB - The identification of the breast cancer susceptibility genes BRCA1 and BRCA2 a few years ago has been greeted with great excitement and has raised hopes that they might illuminate the common mechanisms of this disease. Today we have to recognize that these expectations remain unfulfilled. Mutations in BRCA1 and BRCA2 account only for a relatively small proportion of breast cancers, even within the group of familiar clusters, they seem to be virtually non-existing in sporadic breast cancers. A substantial proportion of familiar breast cancer clusters has failed to provide evidence for an association with mutations in either BRCA1 or BRCA2, thus we have to look forward to the identification of additional breast cancer susceptibility genes. What has been most disappointing is that the mutation status of BRCA1/2 can provide only limited information for cancer risk. Initial assessments had indicated a risk of close to 90% for mutation carriers to develop breast cancer until age 75 - a value that turned out to be restricted to high-risk families in which the BRCA1 and BRCA2 genes had been genomically mapped. In unselected clusters the risk appears much lower, some estimates suggest less than 40%. Both BRCA1 and BRCA2 large encode proteins that appear to have a plethora of functions, with a conspicuous association to DNA repair and DNA recombination, and probably transcription activation. Defects in DNA repair can result in cancer predisposition syndromes and are recognized as being instrumental in cancer progression. Central questions have remained unanswered: What is the function of damaged BRCA1 and BRCA2 genes in breast cancer risk? What is the basis of large variations of risk conferred to the patients by identical mutations? How can the predictive value of mutation surveys be increased? PMID- 11734331 TI - Inhibitory effects of cinnamaldehyde on 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone-induced lung carcinogenesis in rasH2 mice. AB - Previously we reported a lack of modification by cinnamaldehyde (CNMA) of development of lung proliferative lesions induced by urethane in CB6F1-TgHras2 (rasH2) mice. In the present study, we re-evaluated CNMA effects using the same rasH2 strain and non-transgenic littermates initiated with 4-(methylnitrosamino) 1-(3-pyridyl)-1-butanone (NNK). Sixteen mice/strain/sex received intraperitoneal NNK injections at a dose of 3 mg/mouse once a week for 2 weeks followed by free feeding of commercial diet containing 5000 ppm CNMA for 26 weeks. Additional groups were maintained without NNK injection and/or CNMA feeding for 28 weeks. Lung tumors were induced by NNK in both rasH2 and non-Tg males and females at incidence ranging from 63 to 100%. CNMA treatment significantly reduced the combined incidence of adenomas and carcinomas from 86 to 31% in rasH2 males (P<0.05), but no significant influence was evident in females. The multiplicity of NNK-induced lung tumors was also significantly reduced in rasH2 males given CNMA (P<0.01). Similar effects were also observed in non-Tg females given CNMA after NNK initiation. The results of our study strongly indicate that CNMA is capable of inhibiting development of NNK-initiated pulmonary tumorigenesis in rasH2 and non-Tg mice. PMID- 11734332 TI - Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells. AB - Betulinic acid is a triterpene with selective cytotoxicity against melanoma, neuroectodermal and malignant brain tumor cell lines. In this study the betulinic acid activity was evaluated, in comparison with doxorubicin, on different human neoplastic and non-neoplastic cell lines and on proliferating normal lymphocytes. Growth inhibition was evident in all the neoplastic cell lines independently on p53 status and histotype. Antiproliferative activity of betulinic acid was related to a cytotoxic effect on two p53 wild-type and on one p53 mutant cell lines and to a cytostatic effect on one p53 mutant melanoma clone. At the same concentrations, normal cells were unaffected indicating a selective effect of this agent. A cytotoxic activity of doxorubicin was evident on all the tested systems. In vivo experiments, performed on one of these cell lines, confirmed the antineoplastic activity of this drug. These data support further preclinical studies of betulinic acid not confined to melanoma and neuroectodermal tumors independently of p53 status. PMID- 11734333 TI - Induction of apoptosis by cisplatin and its effect on cell cycle-related proteins and cell cycle changes in hepatoma cells. AB - We investigated cisplatin-induced apoptosis and the effects on cell cycle-related proteins and cell cycle changes. Two human hepatoma cell lines, HepG2 (with wild type p53) and Hep3B (with deleted p53), were treated with different concentrations of cisplatin. Cisplatin induced apoptosis in both cell lines as assessed by cell morphology, DNA fragmentation analysis,TdT-mediated dUTP nick end labeling assay and flow cytometry. HepG2 cells were more sensitive to cisplatin than Hep3B. Low-dose cisplatin induced a transient G(1) arrest, S phase block and upregulation of p53 and p21(WAF1/CIP1) expression in HepG2, but not in Hep3B cells. With cisplatin at a high dose, both cell lines underwent apoptosis that was accompanied by downregulation of p27(KIP1) and Bcl-x(L). In HepG2, upregulation of p53 and p21(WAF1/CIP1) was observed before apoptosis occurred, suggesting that cisplatin-induced apoptosis in HepG2 might be p53-dependent. Expression of Fas was also increased following cisplatin treatment in HepG2. However, there was no induction of p53, p21(WAF1/CIP1) and Fas observed in Hep3B cells. In conclusion, cisplatin induced apoptosis in hepatoma cells via both p53 dependent and -independent pathways. PMID- 11734334 TI - The preferential induction of apoptosis in multidrug-resistant KB cells by 5 fluorouracil. AB - It has been previously been shown that multidrug resistance may be associated with biochemical changes which increase the sensitivity of resistant cells to the induction of apoptosis by certain agents. We have shown here that 48 h exposure to 5-fluorouracil (5-FU) induces both a significantly greater proportion of apoptotic cells and much greater cleavage of the apoptosis-related protein poly (ADP-ribose)-polymerase in the multidrug-resistant (MDR) carcinoma cell line, KB A1, than in corresponding drug-sensitive control KB-3.1 cells. Exposure to 5-FU also reduced the level of the anti-apoptotic protein, protein kinase B, in the MDR cells, but not in the control cells. PMID- 11734335 TI - Prevention of liver metastasis of human colon cancer by selective matrix metalloproteinase inhibitor MMI-166. AB - MMI-166 is a selective inhibitor of matrix metalloproteinase (MMP)-2 and MMP-9. Mice implanted a human colon cancer orthotopically received 200 mg/kg of MMI-166 orally for 5 weeks. Gelatin zymography demonstrated that the administration of MMI-166 remarkably decreased the active MMP-2 expression. Histological examination revealed that MMI-166 showed prominent effect on reduction of the invasive feature of the cancer cells and showed inhibitory effect on tumor vasculature, resulting in the significant decrease of microvessel density of the implanted tumor and liver metastasis compared with the control group. Conclusively, MMI-166 is a potent antiangiogenic oral agent for a human colon cancer. PMID- 11734336 TI - Caffeic acid phenethyl ester inhibits nitric oxide synthase gene expression and enzyme activity. AB - Since nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been known to be involved in inflammatory and autoimmune-mediated tissue destruction, modulation of NO synthesis or action represents a new approach to the treatment of inflammatory and autoimmune diseases. Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, has been identified to show anti-inflammatory, anti-viral and anti-cancer activities. The present study, therefore, examined effects of CAPE on iNOS expression and activity of iNOS enzyme itself. Treatment of RAW 264.7 cells with CAPE significantly inhibited NO production and iNOS protein expression induced by lipopolysaccharide (LPS) plus interferon-gamma (IFN-gamma). CAPE also inhibited iNOS mRNA expression and nuclear factor-kappa B (NF-kappaB) binding activity in a concentration-dependent manner. Furthermore, transfection of RAW 264.7 cells with iNOS promoter linked to a chloramphenicol acetyltransferase reporter gene, revealed that CAPE inhibited the iNOS promoter activity induced by LPS plus IFN-gamma through the NF-kappaB sites of the iNOS promoter. In addition, CAPE directly interfered with the catalytic activity of murine recombinant iNOS enzyme. These results suggest that CAPE may exert its anti-inflammatory effect by inhibiting the iNOS gene expression at the transcriptional level through the suppression of NF-kappaB activation, and by directly inhibiting the catalytic activity of iNOS. PMID- 11734337 TI - Analysis of stromal-epithelial interactions in prostate cancer identifies PTPCAAX2 as a potential oncogene. AB - A PCR-based subtractive hybridisation technique was used to identify genes involved in stromal-epithelial interactions in prostate cancer. Eight genes were identified as being differentially expressed in benign prostatic fibroblast cells after stimulation with tumourigenic LNCaP conditioned media. One of these genes, protein tyrosine phosphatase CAAX2 (PTPCAAX2; also described as PTP4A and OV-1), has recently been shown to be oncogenic in hamster pancreatic epithelial cells. We show that PTPCAAX2 expression is up-regulated 4-fold in benign prostatic fibroblast cells 24 h after stimulation with LNCaP conditioned media and up regulated 9-fold in prostatic tumour fibroblast cells. PTPCAAX2 overexpression was also detected in both androgen-dependent and androgen-independent prostate cancer cell lines and prostate tumour tissue, as determined by RT-PCR analysis and in situ hybridisation. These observations of PTPCAAX2 overexpression in prostate tumour cells and tissue suggest that PTPCAAX2 may potentially function as an oncogene in prostate cancer. PMID- 11734338 TI - Induced transcriptional expression of calcium-binding protein S100A1 and S100A10 genes in human renal cell carcinoma. AB - Expression of 16 S100 family calcium-binding protein genes was evaluated by PCR in ten human tissue cDNA libraries. Six to 12 S100 genes were expressed in a tissue-specific manner. Then, the expression in the surgically resected renal cell carcinoma (RCC) and a cultured RCC cell line was studied by RT-PCR. Although eight to nine S100 genes were transcribed in the normal kidney library and non cancerous part of resected kidney tumors, S100A1 and S100A10 genes were not expressed. However, these genes were newly expressed in the RCC lesions (n=7) and the RCC cell line, indicating that expression of S100A1 and S100A10 genes is accompanied by RCC. PMID- 11734339 TI - Mechanism(s) of turmeric-mediated protective effects against benzo(a)pyrene derived DNA adducts. AB - The effects of turmeric feeding before and after benzo(a)pyrene [B(a)P] exposure on the levels of B(a)P-derived DNA adducts were studied in tissues of Swiss mice employing (32)P-postlabelling analysis. A reduction in the levels of B(a)P derived DNA adducts in liver, lung, and forestomach was observed in animals pre treated with 0.2 or 1% turmeric diet and exposed to B(a)P by oral intubation when compared to animals receiving standard laboratory diet and B(a)P. The observed decrease was not due to dilution caused by nascent DNA synthesis. Comparative evaluation of levels of B(a)P-derived DNA adducts in tissues of animals shifted to 0.2 or 1% turmeric diet after 24 h of oral intubation of B(a)P with those continued on standard laboratory diet did not suggest enhanced disappearance/repair of B(a)P-derived DNA adducts due to exposure to turmeric. Further, pre-treatment of mice with 1% turmeric diet significantly reduced the B(a)P-induced increase in activity of cytochrome P450 (CYP450) isozymes CYP 1A1 and 1A2 in liver, lung, and forestomach of mice. In addition, hepatic glutathione S-transferase (GST) was found to be elevated in turmeric pre-treated mice. Thus turmeric-mediated decrease in induction of phase-I enzymes in liver, lung, and forestomach of mice and enhancement of hepatic GST appear to play an important role in reducing the B(a)P-induced DNA damage in target and non-target tissues. PMID- 11734340 TI - Inhibition of cell transformation by sulindac sulfide is confined to specific oncogenic pathways. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) have been shown to reduce the risk of colorectal cancer (CRC). They are also known to induce the regression of colorectal adenomas, which are precursors to CRC. Despite these evidences, the exact mechanism by which NSAIDs exerts its anti-oncogenic effect is not completely understood. Using a focus formation assay, here we show that sulindac sulfide, a NSAID, specifically inhibits cell transformation mediated by oncogenic Ha-Ras, but not by other established oncogene products such as v-Src, Galpha12, and Galpha13. Our results suggest that the ability of sulindac sulfide to suppress transformation is confined to specific oncogenic pathways. Further studies of the sulindac-resistant oncogenic pathways may lead to identification of novel therapeutic agents that are effective in the prevention or treatment of CRC. PMID- 11734341 TI - Enhancement of tumorigenic, metastatic and in vitro invasive capacity of rat mammary tumor cells by transforming growth factor-beta. AB - We investigated the effects of transforming growth factor-beta (TGF-beta) on biological behavior of a weakly malignant rat mammary carcinoma ER-1 cell line. TGF-beta enhanced the tumorigenic and metastatic capacity of ER-1 cells and their in vitro invasiveness to rat mesothelial and endothelial cell. Further cell biological analysis indicated that the increased invasive and metastatic capacity of ER-1 cells by TGF-beta was due to the increase in cell motility and adhesion to the mesothelial and endothelial cell monolayers. Thus, it is suggested that TGF-beta acts on ER-1 cells as a progression-enhancing factor which stimulates their adhesive and motile activities. PMID- 11734343 TI - Experimental investigation of the effect orifice shape and fluid pressure has on high aspect ratio cross-sectional jet behaviour. AB - Prevention of major disasters such as Piper Alpha is a concern of oil and gas companies when commissioning a new offshore superstructure. Safety studies are undertaken to identify potential major hazards, risks to personnel and that sufficient precautions have been employed to minimise these. Such an assessment will also include the consideration of the protection from gas leaks such as the optimum positions of gas leak detectors and startup safety procedures after a leak. This requires a comprehensive knowledge of the behaviour of the leaking hydrocarbons as they emerge from the leak into the area of concern. Such leaks are most likely to emanate from a high aspect ratio cross-sectional curved slot in a pipeline. This paper challenges the conventional view that it is sufficient to model such leaks as axisymmetric jets. This paper is therefore concerned with an experimental study carried out on a series of more realistic high aspect ratio cross-sectional jets issuing from a flange orifice. Both high quality photographs in both planes of the jets and some quantitative pressure data is examined for a high aspect ratio cross-sectional jet of air at pressures up to 4.136bar. The effect of changing aspect ratio, fluid pressure and orifice shape will be discussed and put into context with regard to how this relates to offshore analysis studies. PMID- 11734344 TI - A 12-year history of hazardous materials incidents in Chester County, Pennsylvania. AB - In compliance with the provisions of the Emergency Planning and Community Right to Know Act (EPCRA) of 1986, Chester County, Pennsylvania has collected information about all reported chemical releases to the environment. This paper presents an analysis of the patterns of these releases, including their composition, type, location, frequency, time and level of emergency response, over the last 12 years (1987-1999). Hydrocarbon fuels are the most frequently spilled compounds, but several extremely hazardous chemicals including sulfuric acid, chlorine, and ammonia have also been released to the environment. The patterns of hazardous materials releases in Chester County are similar to those reported in other databases at all levels: local, state, federal and international. PMID- 11734345 TI - Residue levels of DDE and PCBs in the blood serum of women in the Port Said region of Egypt. AB - An investigation was conducted to detect residues of 1,1-dichloro-2,2-bis(p chlorophenyl) ethylene (DDE) and polychlorinated biphenyls (PCBs) in blood serum samples collected from a cohort of fasting females attending the health insurance outpatient clinic at Port Said between July 1999 and July 2000. Females involved in the study included 43 females diagnosed with invasive adenocarcinoma of the breast, 21 female suffering benign breast disease, and 11 normal healthy females. Serum was separated and its contents of DDE and PCBs were extracted and determined, using gas chromatography, equipped with electron capture detector. Mean residues of DDE detected in the three examined groups of females were 41+/ 5.2, 48+/-6.2 and 31+/-2.5ng/g for breast cancer cases, benign breast disease cases and controls, respectively, indicating some significantly less residues in blood serum of control females. While PCBs residues detected were 54+/-17, 59+/ 23 and 61+/-21ng/g, for the three groups, respectively. Residues of DDE detected in all females alike in the present study are about 15 times higher than residues detected in Canada and The Netherlands. PMID- 11734346 TI - Degradation of carbofuran by using ozone, UV radiation and advanced oxidation processes. AB - The degradation of carbofuran (2,3-dihydro-2,2-dimethylbenzofuran-7-yl methylcarbamate), a frequently used carbamate derivative pesticide that is considered a priority pollutant, is carried out in batch reactors by means of single oxidants: ozone, UV radiation and Fenton's reagent; and by the advanced oxidation processes (AOPs) constituted by combinations of ozone plus UV radiation, UV radiation plus H(2)O(2), and UV radiation plus Fenton's reagent (photo-Fenton system). For all these reactions, the apparent pseudo-first-order rate constants are evaluated in order to compare the efficiency of each process. In addition and by means of a competition kinetic model, the rate constants for the reaction of carbofuran with ozone and with hydroxyl radicals are also determined. The improvement in the decomposition levels of carbofuran reached by the combined processes in relation to the single oxidants, due to the generation of the very reactive hydroxyl radicals, is also established in every process. For the oxidant concentrations applied, the most effective process in removing carbofuran was the photo-Fenton system. PMID- 11734347 TI - Solidification of waste steel foundry dust with Portland cement. AB - Properties of mixtures of Portland cements and "waste steel foundry dust (WSFD)" from Czech steel works were examined. It was found that WSFDs are formed by microporous clusters of spherical particles of iron oxides, ranging in size from 10 to 100 nm. The content of ZnO in WSFD affects markedly the hydration of cement + WSFD mixtures both in solidification time and strength development. Properties of WSFD are very similar to those of very fine silicon flue dusts formed during Si and FeSi production. Cement + WSFD mixtures show long term strength stability and low heavy metals leaching even at WSFD content of 70-80wt.% The above results document the perspectives of WSFD solidification, disposal and the use of this waste material as a new additive to building materials. PMID- 11734348 TI - Removal of nitrate from water by electroreduction and electrocoagulation. AB - The aim of this work was to investigate the feasibility of the removal of nitrate from water by applying electrochemical methods such as electroreduction and electrocoagulation. In electroreduction, removal of nitrate to an allowable concentration has been accomplished at the pH range of 5-7 with energy consumption value of 1 x 10(-3) kWh g(-1). In electrocoagulation, an allowable concentration of nitrate has been achieved at the pH range of 9-11 with energy consumption value of 0.5 x 10(-4) kWh g(-1). Full removal of nitrate was also possible but with higher energy consumptions for these two methods. PMID- 11734353 TI - Orexins/hypocretins excite basal forebrain cholinergic neurones. AB - The orexins (orexin A and B, also known as hypocretin 1 and 2) are two recently identified neuropeptides (de Lecea et al., 1998; Sakurai et al., 1998) which are importantly implicated in the control of wakefulness (for reviews see Hungs and Mignot, 2001; van den Pol, 2000; Willie et al., 2001 ). Indeed, alteration in these peptides' precursor, their receptors or the hypothalamic neurones that produce them leads to the sleep disorder narcolepsy (Chemelli et al., 1999; Lin et al., 1999; Peyron et al., 2000; Thannickal et al., 2000). The mechanisms by which the orexins modulate wakefulness, however, are still unclear. Their presence in fibres coursing from the hypothalamus (Peyron et al., 1998) up to the preoptic area (POA) and basal forebrain (BF) suggests that they might influence the important sleep and waking neural systems situated there (Jones, 2000). The present study, performed in rat brain slices, demonstrates, however, that the orexins have no effect on the GABA sleep-promoting neurones of the POA, whereas they have a strong and direct excitatory effect on the cholinergic neurones of the contiguous BF. In addition, by comparing the effects of orexin A and B we demonstrate here that orexins' action depends upon orexin type 2 receptors (OX(2)), which are those lacking in narcoleptic dogs (Lin et al., 1999). These results suggest that the orexins excite cholinergic neurones that release acetylcholine in the cerebral cortex and thereby contribute to the cortical activation associated with wakefulness. PMID- 11734354 TI - Coordinate expression of Fgf8, Otx2, Bmp4, and Shh in the rostral prosencephalon during development of the telencephalic and optic vesicles. AB - Previous studies suggest that Fgf8 has a key role in regulating vertebrate development. In the rostral head of the embryonic chicken, there are increasing numbers of separate Fgf8 domains; these are present in tissues that appear to have previously expressed Otx2. As Fgf8 expression becomes established, Otx2 expression weakens, but remains in cells abutting the Fgf8 expression domain. These Fgf8 expression domains are closely associated with tissues expressing Bmp4 and Shh. Based on analogy with the embryonic limb, we suggest that Fgf8, Bmp4 and Shh function together in patterning regions of the embryonic head. Gene expression changes are particularly prominent in 14-21 somite stage embryos in the rostral forebrain, during early morphogenesis of the telencephalic and optic vesicles, when several new interfaces of Fgf8, Bmp4 and Shh are generated. To gain insights into the functions of fibroblast growth factor 8 (FGF8) in the embryonic forebrain, we studied the effects of implanting beads containing this protein in the dorsal prosencephalon of embryonic day 2 chicken embryos. Ectopic FGF8 had profound effects on morphogenesis of the telencephalic and optic vesicles. It disrupted formation of the optic stalk and caused a transformation of the pigment epithelium into neural retina. Within the telencephalon, FGF8 beads frequently induced a sulcus that had features of an ectopic rostral midline. The sulcus separated the telencephalon into rostral and caudal vesicles. Furthermore, we present evidence that FGF8 can regulate regionalization of the prosencephalon through inhibition of Otx2 and Emx2 expression. Thus, these experiments provide evidence that FGF8 can regulate both morphogenesis and patterning of the rostral prosencephalon (telencephalic and optic vesicles). FGF8 beads can induce midline properties (e.g. a sulcus) and can modulate the specification and differentiation of adjacent tissues. We suggest that some of these effects are through regulating the expression of homeobox genes (Otx2 and Emx2) that are known to participate in forebrain patterning. PMID- 11734355 TI - Ntab, a novel non-coding RNA abundantly expressed in rat brain. AB - We have identified a novel transcript that is abundantly and specifically expressed in both the adult and developing rat CNS. Within the full-length cDNA sequence we were unable to identify a clear open reading frame. Moreover, we were unable to detect any protein product derived from the full-length cDNA sequence using an in vitro translation assay. Therefore, we suggest this gene is one of a growing number of non-coding mRNA-like RNA transcripts that exert their cellular functions directly as an RNA. We have named this novel gene Ntab for non-coding transcript abundantly expressed in brain (accession number AY035551). In addition, in some regions of the brain we find evidence for RNA accumulation in cellular processes at some distance from the soma. These findings suggest that Ntab is actively transported and may function within cellular processes. Since Ntab is a targeted non-coding RNA, such cellular functions could include the targeting and/or regulation of localised translation of other mRNA species. PMID- 11734356 TI - Normotopic and heterotopic cortical representations of mystacial vibrissae in rats with subcortical band heterotopia. AB - The tish rat is a neurological mutant exhibiting bilateral cortical heterotopia similar to those found in certain epileptic patients. Previous work has shown that thalamocortical fibers originating in the ventroposteromedial nucleus, which in normal animals segregate as 'barrel' representations for individual whiskers, terminate in both normotopic and heterotopic areas of the tish cortex (Schottler et al., 1998). Thalamocortical innervation terminates as barrels in layer IV and diffusely in layer VI of the normotopic area. Discrete patches of terminals are also observed in the underlying heterotopic area suggesting that representations of individual vibrissa may be present in the heterotopic somatosensory areas. The present study examines this issue by investigating the organization of the vibrissal somatosensory system in the tish cortex. Staining for cytochrome oxidase or Nissl substance reveals a normal complement of vibrissal barrels in the normotopic area of the tish cortex. Dense patches of cytochrome oxidase staining are also found in the underlying lateral portions of the heterotopic area (i.e. the same area that is innervated by the ventroposteromedial nucleus). Injections of retrograde tracers into vibrissal areas of either the normotopic or heterotopic area produce topographically organized labeling of neurons restricted to one or a small number of barreloids within the ventroposteromedial nucleus of the thalamus. Physical stimulation of a single whisker (D3 or E3) elicits enhanced uptake of [(14)C]2-deoxyglucose in restricted zones of both the normotopic and heterotopic areas, demonstrating that single whisker stimulation can increase functional activity in both normotopic and heterotopic neurons. These findings indicate that the barrels are intact in the normotopic area and are most consistent with the hypothesis that at least some of the individual vibrissae are 'dually' represented in normotopic and heterotopic positions in the primary somatosensory areas of the tish cortex. PMID- 11734357 TI - Reversal of cognitive deficit of apolipoprotein E knockout mice after repeated exposure to a common environmental experience. AB - This study tests the hypothesis that a history of common stressful experiences further promotes the cognitive deficit of apolipoprotein E (apoE)-knockout mice, an animal model to study aspects of Alzheimer's disease. In experiment 1, apoE knockout and wild-type mice were repeatedly subjected to an environmental challenge (i.e. exposure to rats) and the effect was monitored on Morris water maze performance. Naive apoE-knockout mice were impaired, but surprisingly after rat stress their water maze performance improved and switched to a goal-directed search strategy. Rat stress induced in wild-type mice spatial learning deficits and an inefficient search strategy. Swim ability was not affected by rat stress and under basal conditions measures for locomotion and anxiety were similar for both genotypes. In experiments 2 and 3, we found that the rat stress paradigm attenuated the elevation of basal and stress-induced corticosterone concentrations in the apoE-knockout mice towards concentrations observed in wild type mice. The expression of hippocampal mineralocorticoid and glucocorticoid receptor mRNA was similar in both genotypes, but in response to rat stress, the level of glucocorticoid receptor mRNA increased selectively in the CA1 pyramidal field. In conclusion, repeated exposure to a common environmental experience did abolish and reverse the difference in cognitive performance and corticosterone concentrations of apoE-knockout and wild-type mice. PMID- 11734358 TI - Activity-dependent changes of tissue resistivity in the CA1 region in vivo are layer-specific: modulation of evoked potentials. AB - We have investigated the spatial map of tissue resistivity across CA1 layers in vivo, its modifications during repetitive orthodromic activity, and the influence of this factor on the shaping of population spikes. Measurement of tissue resistance was made by a high-spatial-resolution three-electrode method. A computer network of equivalent resistors aided theoretical analysis. Tissue resistivity was homogeneous within the basal and apical dendritic trees (260+/ 4.5 and 287+/-2.6 Omega cm, respectively). In the stratum pyramidale we found a sharply delimited high-resistivity (643+/-35 Omega cm) band approximately 20 microm wide. Resistivity in slices was approximately 30% higher than in vivo. Computer analysis indicated that the high-resistance somatic layer has a strong influence on the somatic and proximal dendritic contribution to the shaping of population spikes, and reduces volume propagation of currents between dendritic trees. Repetitive orthodromic activation at the theta frequency (4-5 Hz, 20-30 s) caused a stereotyped cycle of field potentials and layer-specific changes of resistivity. Initially (approximately 10 s), long-lasting field excitatory postsynaptic potentials and multiple somatodendritic population spikes developed, and resistivity gradually increased in all layers at a similar rate (period average: 11%). Subsequently, the long-lasting field excitatory synaptic potential subsided and dendritic spike generators were strongly reduced, but multiple somatic spikes remained. Concurrently, the resistivity reached a plateau in all dendritic layers but continued to increase in the somatic layer for about 10-15 s (20% average and up to 50% maximum). Recovery required approximately 60 s. The orthodromic somatic population spike increased variably during stimulation (up to 60%). Using local resistivity changes for correction, supernormal increments of the population spikes were offset, but not totally, uncovering several sub- and supernormal phases that were partially related to changes in dendritic population spike. These resistivity-independent modulations of the somatic population spike are caused by variable volume spread from dendritic spike currents and changed somatic contribution of firing units. This report demonstrates that the strong heterogeneity in the stratum pyramidale is an important factor shaping and modulating the population spike. The different regional rates of resistivity variation force the independent correction of local evoked potentials. We show that not doing so may cause bulk errors in the interpretation of, for instance, field potential ratios widely used to measure the population excitability. The present results underscore the importance of checking variations in recording conditions, which are inherent in most experimental protocols. PMID- 11734359 TI - Morphological and functional properties of rat dentate granule cells after adrenalectomy. AB - After complete adrenalectomy, part of the granule cells in the dentate gyrus undergo apoptosis. Findings on morphological changes in non-apoptotic granule cells, though, have been equivocal. In the present study we examined the dendritic trees of dentate granule cells 7 days after adrenalectomy or sham operation, and tested the hypothesis that changes in dendritic trees have considerable consequences for ionic currents, as measured in the soma with whole cell recording. For the latter, we focussed on voltage-gated calcium currents, which are partly generated in distal dendrites. All cells were passively filled with a fluorescent dye via the patch pipette while recording calcium currents; subsequently the cells were three-dimensionally reconstructed with the use of a confocal microscope. In sham-operated rats, dendritic trees of cells with a soma located in the inner part of the granule cell layer (facing the hilus) were significantly smaller than trees of cells located in the outer part of the layer. Neurons from rats that had extremely low (undetectable-0.3 microg/dl) circulating levels of corticosterone displayed very small and simple dendritic trees compared to cells from adrenalectomized rats that still had residual levels of corticosterone (0.6-1.0 microg/dl), regardless of the location of their soma. Despite the observation that simple dendritic trees were seen in rats where corticosterone was extremely low, the whole cell calcium current amplitude recorded from the soma of these cells was not reduced compared to the remaining cells from adrenalectomized or sham-operated rats. Our data indicate that in the absence of corticosterone dendritic trees of dentate granule cells display atrophy but that this does not necessarily reduce ionic currents measured in the soma. PMID- 11734360 TI - Additive effect of glial cell line-derived neurotrophic factor and neurotrophin 4/5 on rat fetal nigral explant cultures. AB - Transplantation of embryonic dopaminergic neurons is an experimental therapy for Parkinson's disease, but limited tissue availability and suboptimal survival of grafted dopaminergic neurons impede more widespread clinical application. Glial cell line-derived neurotrophic factor (GDNF) and neurotrophin-4/5 (NT-4/5) exert neurotrophic effects on dopaminergic neurons via different receptor systems. In this study, we investigated possible additive or synergistic effects of combined GDNF and NT-4/5 treatment on rat embryonic (embryonic day 14) nigral explant cultures grown for 8 days. Contrary to cultures treated with GDNF alone, cultures exposed to NT-4/5 and GDNF+NT-4/5 were significantly larger than controls (1.6- and 2.0-fold, respectively) and contained significantly more protein (1.6-fold). Treatment with GDNF, NT-4/5 and GDNF+NT-4/5 significantly increased dopamine levels in the culture medium by 1.5-, 2.5- and 4.7-fold, respectively, compared to control levels, and the numbers of surviving tyrosine hydroxylase immunoreactive neurons increased by 1.7-, 2.1-, and 3.4-fold, respectively. Tyrosine hydroxylase enzyme activity was moderately increased in all treatment groups compared to controls. Counts of nigral neurons containing the calcium binding protein, calbindin-D28k, revealed a marked increase in these cells by combined GDNF and NT-4/5 treatment. Western blots for neuron-specific enolase suggested an enhanced neuronal content in cultures after combination treatment, whereas the expression of glial markers was unaffected. The release of lactate dehydrogenase into the culture medium was significantly reduced for GDNF+NT-4/5 treated cultures only. These results indicate that combined treatment with GDNF and NT4/5 may be beneficial for embryonic nigral donor tissue either prior to, or in conjunction with, intrastriatal transplantation in Parkinson's disease. PMID- 11734361 TI - Perineuronal nets in the rhesus monkey and human basal forebrain including basal ganglia. AB - Perineuronal nets of extracellular matrix have been shown to characterize the microenvironment of individual neurons and the chemoarchitecture of brain regions such as basal forebrain nuclei. Previous work has also demonstrated that neurons in the human cerebral cortex ensheathed by perineuronal nets rarely undergo cytoskeletal changes in Alzheimer's disease, suggesting a neuroprotective effect of extracellular matrix components. It is not known, however, whether or not perineuronal nets are absent in the microenvironment of the cholinergic basal forebrain neurons that are involved early in the cascade of neurodegeneration in humans. Therefore, the present study was undertaken to examine the distribution patterns of perineuronal nets in the basal forebrain of the higher primates, rhesus monkey and human. Cytochemical staining was performed with the lectin Wisteria floribunda agglutinin and a polyclonal antibody to core proteins of chondroitin sulfate proteoglycans in the perfusion-fixed tissue of rhesus monkeys. In human brains, perineuronal nets were only stained with the immunoreaction for chondroitin sulfate proteoglycans. The results showed similar characteristics in distribution patterns of perineuronal nets in the medial septum, the diagonal band of Broca, the basal nucleus of Meynert (Ch1-Ch4), the lateral septum, the caudate-putamen, and the globus pallidus in both species. Double-labelling revealed that the vast majority of cholinergic neurons, labelled either with antibodies to choline acetyltransferase or the low-affinity neurotrophin receptor p75(NTR), were not ensheathed by perineuronal nets. A small subpopulation of net-associated neurons in close proximity to or intermingled with cholinergic neurons of the Ch1-Ch4 cell groups was found to be immunoreactive for parvalbumin. In the caudate-putamen, a large number of the parvalbumin-positive neurons were surrounded by perineuronal nets, whereas in the external and internal segments of the globus pallidus the coincidence of both markers was nearly complete. The study demonstrates that perineuronal nets of extracellular matrix are associated with different types of non-cholinergic neurons in the primate basal forebrain. The absence of nets around cholinergic basal forebrain neurons may be related to their slow modulatory activity but may also contribute to their susceptibility to degeneration in Alzheimer's disease. PMID- 11734362 TI - Selective immunolesioning of cholinergic neurons in nucleus basalis magnocellularis impairs prepulse inhibition of acoustic startle. AB - Information processing and attentional abnormalities are prominent in neuropsychiatric disorders. Since the cholinergic neurons located in the nucleus basalis magnocellularis have been shown to be involved in attentional performance and information processing, recent efforts to analyze the significance of the basal forebrain in the context of schizophrenia have focused on this nucleus and its projections to the cerebral cortex. We report here that bilateral selective immunolesioning of the cholinergic neurons in the nucleus basalis magnocellularis is followed by significant deficits in sensorimotor gating measured by prepulse inhibition of the startle reflex in adult rats. This behavioral approach is used in both humans and rodents and has been proposed as a valuable model contributing to the understanding of the neurobiological substrates of schizophrenia. The disruption of prepulse inhibition persisted over repeated testing. The selective lesions were induced by bilateral intraparenchymal infusions of 192 IgG saporin at a concentration having minimal diffusion into adjacent nuclei of the basal forebrain. The infusions were followed by extensive loss of choline acetyltransferase-immunopositive neurons. Our results show that the cholinergic neurons of the nucleus basalis magnocellularis represent a critical station of the startle gating circuitry and suggest that dysfunction of these neurons may result in impaired sensorimotor gating characteristic of schizophrenia. PMID- 11734363 TI - Distribution and amino acid content of enkephalin-immunoreactive inputs onto juxtacellularly labelled bulbospinal barosensitive neurons in rat rostral ventrolateral medulla. AB - The activity of bulbospinal (presympathetic) vasomotor neurons of the rostral ventrolateral medulla is modulated pre- and postsynaptically by exogenously applied opioid agonists. To determine whether these neurons receive direct opioid inputs, we examined the relationship between bulbospinal barosensitive neurons and nerve terminals immunoreactive for enkephalin in the rostral ventrolateral medulla of rats. By light microscopy, we mapped the distribution of close appositions by enkephalin-immunoreactive varicosities on 10 bulbospinal barosensitive neurons labelled in vivo with biotinamide. We also examined four labelled neurons ultrastructurally for synapses by enkephalin-immunoreactive terminals and determined with post-embedding immunogold labelling whether these enkephalin-positive terminals contained amino acids. Enkephalin-immunoreactive varicosities closely apposed all bulbospinal barosensitive neurons. Maps of the dendritic distribution of appositions indicated that fast-conducting bulbospinal barosensitive neurons with myelinated axons (conduction velocity >3 m/s; n=3) received many appositions (up to 470/neuron); and slowly conducting neurons with unmyelinated axons (conduction velocity <0.90 m/s; n=3), substantially fewer. Ultrastructural analysis of three fast- and one slowly conducting bulbospinal barosensitive neurons revealed numerous synapses from enkephalin-immunoreactive terminals on cell bodies and dendrites. Enkephalin-positive terminals synapsing on bulbospinal barosensitive neurons contained one or more amino acid: GABA+glycine, glutamate alone or GABA+glutamate. Enkephalin-immunoreactive terminals located near biotinamide-labelled cells contained a similar variety of amino acids. In summary, enkephalin-immunoreactive terminals in the rostral ventrolateral medulla densely innervate lightly myelinated presympathetic neurons and more sparsely those with unmyelinated axons. Enkephalin is present in both excitatory (glutamate-immunoreactive) and inhibitory (GABA- and/or glycine immunoreactive) terminals. The data suggest that endogenous enkephalin inhibits amino acid release from terminals that innervate bulbospinal barosensitive neurons of the rostral ventrolateral medulla. PMID- 11734365 TI - Testosterone and nerve growth factor have distinct but interacting effects on structure and neurotransmitter expression of adult pelvic ganglion cells in vitro. AB - Circulating testosterone has potent effects on the structure and function of many pelvic ganglion cells in adult rats in vivo. However not all androgen-sensitive pelvic neurones possess androgen receptors and testosterone effects may therefore be indirect, by an action on the target organs. Here we have examined if testosterone influences neuronal structure in vitro in pelvic ganglion cells cultured from adult male rats. We have also used multiple label immunofluorescence to monitor the expression of transmitter-synthesising enzymes and peptides under various culture conditions. Testosterone was a more potent stimulant of noradrenergic soma growth in culture than nerve growth factor. Whereas nerve growth factor increased the number, branching and length of neurites, testosterone stimulated growth of a small number of very short processes, each of which bore numerous short protrusions. Testosterone also impeded the longer neurite growth induced by nerve growth factor. Many pelvic ganglion cells altered their expression of transmitters/neuropeptides under different culture conditions. In particular, under control conditions or during nerve growth factor treatment, vasoactive intestinal peptide was up-regulated in noradrenergic and cholinergic neurones; testosterone impeded this up-regulation in noradrenergic neurones. Choline acetyltransferase immunoreactivity could only be visualised when nerve growth factor was present in the cultures, and cholinergic neurones showed less neurite outgrowth than noradrenergic neurones under all culture conditions. Nerve growth factor did not stimulate levels of this enzyme as strongly if testosterone was present. This study has shown that testosterone has potent effects on the structure of many pelvic ganglion cells in vitro. It is possible that these effects are mediated indirectly, e.g. by stimulating glial-derived substances, however our results suggest that the effects are not mediated by nerve growth factor. The results also show that testosterone influences some of the actions of nerve growth factor, suggesting that there may be complex interactions between steroid signalling and neurotrophic factors in maintaining neuronal structure and function in vivo. PMID- 11734364 TI - Nociceptin-induced outward current in substantia gelatinosa neurones of the adult rat spinal cord. AB - Nociceptin (NOC), also known as orphanin FQ, is a newly discovered endogenous ligand for the opioid receptor-like1 (ORL1) receptor. Although NOC has been shown to modulate nociceptive transmission, mechanisms for this action are still unknown. In the present study, actions of NOC on substantia gelatinosa (SG) neurones were examined in adult rat spinal cord slice preparations by using the whole-cell patch-clamp technique. NOC at a concentration of 1 microM induced an outward current having an amplitude of 26+/-5 pA (n=68) at a holding potential of -70 mV; this action was dose-dependent with an EC(50) value of 0.23 microM (Hill coefficient: 1.5). The NOC current reversed its polarity at a potential which was close to the equilibrium potential for K(+), as calculated by the Nernst equation (n=4). The NOC current had slope conductances of 0.80+/-0.15 nS and 0.50+/-0.13 nS (n=4) in voltage ranges of -120 to -140 mV and of -60 to -90 mV, respectively. The NOC current was inhibited by Ba(2+) (100 microM; by 56+/-8%, n=4) but not by 4-aminopyridine (4-AP; 1 mM; n=4) and tetraethylammonium (TEA; 5 mM; n=4). The NOC current was not affected by tetrodotoxin (TTX; 1 microM; n=4) and also by a non-specific opioid receptor antagonist, naloxone (1 microM; n=4). When examined using some inhibitors with respect to the ORL1 receptor, the NOC (1 microM) current was depressed in amplitude by a putative NOC precursor product, nocistatin (1 microM; by 18+/-4%, n=6) and also by a non-peptidyl ORL1 receptor antagonist, CompB (1 microM; by 64+/-10%, n=7) without a change in holding currents. On the other hand, a putative ORL1 receptor antagonist, [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin-(1-13)-NH(2) (1 microM; which is a derivative of NOC), by itself induced an outward current (7+/-3 pA, n=8), during which the NOC current was suppressed in amplitude by 56+/-8% (n=8). We conclude that NOC activates in SG neurones a K(+) channel exhibiting a mild inwardly rectification through the activation of ORL1 receptor; this hyperpolarising action of NOC might contribute to at least a part of its antinociceptive effect. PMID- 11734366 TI - Chromaffin cell mitogenesis by neurturin and glial cell line-derived neurotrophic factor. AB - Neurturin and glial cell line-derived neurotrophic factor are novel mitogens for normal adult rat chromaffin cells in vitro. These neurotrophic factors differ from the previously described adult chromaffin cell mitogens, nerve growth factor and basic fibroblast growth factor, in that their effects are potentiated by depolarization and activation of protein kinase C. Neurturin and glial cell line derived neurotrophic factor signal via the receptor tyrosine kinase, ret, but may also act independently of ret. Both depolarization and phorbol esters act synergistically with neurturin to up-regulate ret protein expression in chromaffin cell cultures, suggesting a mechanism for potentiation of mitogenesis. However, a direct role for ret in mitogenesis has not been established. Stimulation by neurturin causes increased phosphorylation of extracellular signal regulated kinases 1 and 2 in cultured chromaffin cells, and mitogenesis is prevented by inhibitors of their phosphorylation. Inhibitors of phosphatidylinositol 3-kinase also prevent mitogenesis. The present findings suggest the hypothesis that neurotrophic factors and neurally derived signals might cooperatively regulate chromaffin cell proliferation in vivo in the rat. In addition, trans-synaptic stimulation might provide a route by which epigenetic factors could influence the development of adrenal medullary hyperplasia in humans with hereditary multiple endocrine neoplasia syndromes 2A and 2B by affecting expression and/or activation of ret. PMID- 11734367 TI - Caspase-3 and the regulation of hypoxic neuronal death by vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) has neurotrophic and neuroprotective as well as angiogenic properties, but the pathways involved in VEGF-mediated neuronal survival have not been identified. We found previously that VEGF protects cultured neural cells from death induced by serum withdrawal or hypoxia via the activation of VEGF-2/fetal liver kinase-1 receptors, phosphatidylinositol 3'-kinase, Akt and nuclear factor-kappa B. We now report that in mouse cortical neuron cultures subjected to hypoxia, the neuroprotective effect of VEGF involves suppression of cell-death pathways mediated by caspase-3. Exposure to hypoxia for 24 h caused the death of 71+/-4% of cultured neurons; this was reduced to 40+/-1% by VEGF (n=3, P<0.005) and to 44+/-1% by the caspase-3 inhibitor benzyloxycarbonyl-DEVD-fluoromethyl ketone (n=3, P<0.005). VEGF inhibited the activation of caspase-3 as measured by the 17-20-kDa caspase-3 cleavage product, and immunolocalization of VEGF and activated caspase-3 showed segregated expression in separate neuronal populations. An antisense, but not sense, oligodeoxyribonucleotide directed against VEGF increased the proportion of neurons expressing activated caspase-3, and correspondingly reduced the viability of hypoxic neurons by 37+/-2% (n=3, P<0.005). These findings suggest that VEGF protects neurons from hypoxic injury by inhibiting the activation of caspase-3, and could therefore act as an endogenous neuroprotective factor in cerebral ischemia. PMID- 11734368 TI - Fourier volume rendering for real time preview of digital reconstructed radiographs: a web-based implementation. AB - Digital reconstructed radiograph (DRR) constitutes an important tool in clinical radio-oncology. In this paper, a Fourier volume rendering (FVR) method is proposed, enabling real time preview of DRR. Important aspects related to re sampling into the frequency domain are examined, leading to results of high quality. Experiments using phantom and clinical data sets are presented, demonstrating the effectiveness of the method for a real time execution. Furthermore, due to the inherent parallelism of the proposed algorithm, its execution time can be further reduced by distributing the required computations to more than one processors. The computational efficiency of the method highlights its suitability for integration into a web-based radiation treatment planning (RTP) system, using a client-server architecture. Thus, platform independent, real time tele-cooperation in clinical radio-oncology can be achieved. PMID- 11734369 TI - Two-phase active contour method for semiautomatic segmentation of the heart and blood vessels from MRI images for 3D visualization. AB - The paper presents an active-contour segmentation method for 2D structures in MR images. The method combines two approaches to active contour segmentation, known as balloons and snakes. This makes the method shape independent and accurate. New anti-tangling features were introduced to improve segmentation of very complex object shapes, e.g. the left ventricle with papillary muscles. The method was applied to segment all large structures in the cardiovascular system and its outcome was used for 3D visualization. PMID- 11734370 TI - Discrimination of an infected brain tumor from a cerebral abscess by combined MR perfusion and diffusion imaging. AB - The aim of this study was to investigate the signal characteristics of the abscess wall and tumor wall on diffusion-weighted and perfusion-weighted images and thus to evaluate the feasibility of using combined MR diffusion and perfusion imaging to differentiate pyogenic cerebral abscess from infected brain tumor. The tumor wall of various types of cystic or necrotic brain tumor was significantly hyperintense relative to that of cerebral abscess wall on both diffusion-weighted images and regional cerebral blood volume maps. Sixteen patients who had cerebral masses with large cystic or necrotic cavities were imaged to generate diffusion weighted images and regional cerebral blood volume maps using single-shot echoplanar imaging (EPI) pulse sequences. Apart from qualitative analysis, apparent diffusion coefficients (ADC) as well as regional cerebral blood volume (rCBV) ratios were calculated from the abscess wall and peripheral tumor wall and comparison was made by using Student's t-test. The tumor wall of various types of cystic or necrotic brain tumor had significantly lower ADCs relative to those of the abscess wall (P<0.005) and thus appeared relatively hyperintense on diffusion weighted images. The mean rCBV ratio relative to normal white matter (2.90+/ 0.62) of the peripheral tumor wall of various types of cystic or necrotic brain tumor were significantly larger than the mean rCBV ratio (0.45+/-0.11) of the pyogenic cerebral abscess wall (P<0.001) by Student's t-test. It is concluded that the combined MR diffusion and perfusion imaging might be capable of differentiating an infected brain tumor from a pyogenic cerebral abscess. PMID- 11734371 TI - Longstanding tectal tumors: proton MR spectroscopy and diffusion MRI findings. AB - Four patients with longstanding tectal tumors are included in this study. The tectal lesion in a neurofibromatosis type 1 (NF1) patient was due to a hamartoma, while others represented gliomas. On spectroscopy, decreased NAA peaks with resultant decreases in the NAA/Cho, and NAA/Cr ratios were observed in two patients. In three patients, on diffusion MRI the tectal tumors were hyperintense on b=1000 s/mm(2) (heavily diffusion-weighted) images. On apparent diffusion coefficient (ADC) maps, they had high signal, and high ADC values (mean=1.20 x 10(-3)mm(2)/s), compared to normal cerebral parenchyma. Diffusion MRI findings suggested that tectal tumors had a relatively high nuclear to cytoplasmic ratio (high-signal on b=1000 s/mm(2) images) in association with a relatively loose intercellular matrix (high ADC values). PMID- 11734372 TI - Texture feature coding method for classification of liver sonography. AB - This paper introduces a new texture analysis method called texture feature coding method (TFCM) for classification of ultrasonic liver images. The TFCM transforms a gray-level image into a feature image in which each pixel is represented by a texture feature number (TFN) coded by TFCM. The TFNs obtained are used to generate a TFN histogram and a TFN co-occurrence matrix (CM), which produces texture feature descriptors for classification. Four conventional texture analysis methods that are gray-level CM, texture spectrum, statistical feature matrix and fractal dimension, are used also to classify liver sonography for comparison. The supervised maximum likelihood (ML) classifiers implemented by different type texture features are applied to discriminate ultrasonic liver images into three disease states that are normal liver, liver hepatitis and cirrhosis. The 30 liver sample images proven by needle biopsy are used to train the ML system that classify on a set of 90 test sample images. Experimental results show that the ML classifier together with TFCM texture features outperforms one with the four conventional methods with respect to classification accuracy. PMID- 11734373 TI - CT findings of non-functioning neuroendocrine pancreatic tumors. AB - Neuroendocrine tumors are rare neoplasms of the pancreas, representing 0.5% of all pancreatic tumors. Approximately, one-third of neuroendocrine tumors are hormonally inactive and called non-functioning neuroendocrine tumors. As these tumors remain clinically silent in their course of growth, they may attain great sizes without causing apparent clinical findings and commonly present in advanced stage. We report three cases of non-functioning neuroendocrine tumors with large sizes and discuss the radiological findings. PMID- 11734374 TI - Benign enlargement of peripheral subarachnoid spaces in twins. AB - A case of benign enlargement of peripheral subarachnoid spaces in twins is described. Computed tomography demonstrated identical findings, namely the enlargement of the sulci mainly in the frontal area bilaterally with slight dilatation of the ventricular system. Literature research did not reveal such a case in twins nor has any association of the situation with any genetic or family predisposition been found. PMID- 11734375 TI - Imaging of an atypical case of congenital toxoplasmosis. AB - In this article we describe the cranial computerized tomography findings of an infant with congenital toxoplasmosis, which was performed because she showed an atypical clinical course consisting of meningitis and multiple cerebral abscesses. In this case the cranial computerized tomography allowed the diagnosis of hydrocephalus and multiple cerebral abscesses, and guided the therapeutic approach. PMID- 11734376 TI - Paraneoplastic limbic encephalitis: case report. AB - Paraneoplastic limbic encephalitis, which is also known as the remote neurological effect of cancer, is a rare neurological paraneoplastic disorder that usually presents with changes in mental status and behavioral abnormalities. Paraneoplastic limbic encephalitis had been diagnosed by clinical, laboratory and pathological studies in the pre-MRI era, but the diagnosis became much easier with MRI. We present MRI pictures of a case of paraneoplastic limbic encephalitis associated with small cell carcinoma of the lung. PMID- 11734377 TI - Side effects and responsibility of medical informatics. AB - Medical informatics systems have the ultimate goal to improve the quality of health care. However, these systems have also the potential to compromise the quality of health care if they are misused, intrinsically faulty or entail unexpected side effects. The purpose of the paper is to discuss examples from the experience of the author, where well-intended medical informatic applications proved to have potentially harmful effects or side effects. It is argued that medical informaticians (MI) have an extended professional responsibility, which covers not only the state of the art technical planning, an implementation of information processing systems in medicine, but also the final result for the patient. In order to discuss professional duties of medical informaticians, a modification of the Software Engineering Code of Ethics and Professional Practice developed by ACM/IEEE is proposed as a guideline. For several examples, these guidelines are used to analyze possible actions and professional responsibility. PMID- 11734378 TI - User requirements specifications: a hierarchical structure covering strategical, tactical and operational requirements. AB - The present study is a long-term review and follow-up on conclusions and recommendations on the role and contents of a User Requirements Specification Document (URD) from a previous in-depth case study. The follow-up comprises a succession of investigations and two case studies to explore the role and contents of a URD within systems development and assessment, thereby gradually extending and refining the original recommendations. Finally, the recommended three-layered structure and approach for a URD were applied in full scale within a real-life project, in which the URD serves as the basis for a Call for Tender. The preparation was entirely user-driven with the aid of a consultant for four man months. The present paper outlines the essence of the approach and the outcome of applying the recommended structure, with numerous examples of the implications. PMID- 11734379 TI - Strategic information management plans: the basis for systematic information management in hospitals. AB - Information management in hospitals is a complex task. In order to reduce complexity, we distinguish strategic, tactical, and operational information management. This is essential, because each of these information management levels views hospital information systems from different perspectives, and therefore uses other methods and tools. Since all these management activities deal only in part with computers, but mainly with human beings and their social behavior, we define a hospital information system as a sociotechnical subsystem of a hospital. Without proper strategic planning it would be a matter of chance, if a hospital information system would fulfil the information strategies goals. In order to support strategic planning and to reduce efforts for creating strategic plans, we propose a practicable structure. PMID- 11734380 TI - Delivering the electronic healthcare record for the 21st century. AB - In spite of over four decades of research into Electronic Healthcare Record Systems, the penetration of records which incorporate more than simply basic information, into the working life of healthcare organisations is relatively small. This paper discusses some of the key impediments to progress including in particular, the lack of application of software engineering methodologies, the absence of usable standards, and the failure to acknowledge the impact of record systems on the healthcare system itself. However, Health Informatics researchers need to be preparing for the next generation of systems which will be triggered by the twin revolutions of the Internet and Genetic Medicine. This next generation of EHCR will be a longitudinal cradle-to-the-grave active record readily accessible and available via the Internet, and which will be linked to clinical protocols and guidelines to drive the delivery of healthcare to the individual citizen. Post-genomic research will unravel the link between genes, disease, treatment and the environment and this information will be used to promote health and individualise care. A number of key research issues are identified which need to be addressed in order to realise the delivery of the next generation of EHCR Systems. PMID- 11734381 TI - Modelling health care processes for eliciting user requirements: a way to link a quality paradigm and clinical information system design. AB - Healthcare institutions are looking at ways to increase their efficiency by reducing costs while providing care services with a high level of safety. Thus, hospital information systems have to support quality improvement objectives. The elicitation of the requirements has to meet users' needs in relation to both the quality (efficacy, safety) and the monitoring of all health care activities (traceability). Information analysts need methods to conceptualise clinical information systems that provide actors with individual benefits and guide behavioural changes. A methodology is proposed to elicit and structure users' requirements using a process-oriented analysis, and it is applied to the blood transfusion process. An object-oriented data model of a process has been defined in order to organise the data dictionary. Although some aspects of activity, such as 'where', 'what else', and 'why' are poorly represented by the data model alone, this method of requirement elicitation fits the dynamic of data input for the process to be traced. A hierarchical representation of hospital activities has to be found for the processes to be interrelated, and for their characteristics to be shared, in order to avoid data redundancy and to fit the gathering of data with the provision of care. PMID- 11734382 TI - Implementing information systems in health care organizations: myths and challenges. AB - Successfully implementing patient care information systems (PCIS) in health care organizations appears to be a difficult task. After critically examining the very notions of 'success' and 'failure', and after discussing the problematic nature of lists of 'critical success- or failure factors', this paper discusses three myths that often hamper implementation processes. Alternative insights are presented, and illustrated with concrete examples. First of all, the implementation of a PCIS is a process of mutual transformation; the organization and the technology transform each other during the implementation process. When this is foreseen, PCIS implementations can be intended strategically to help transform the organization. Second, such a process can only get off the ground when properly supported by both central management and future users. A top down framework for the implementation is crucial to turn user-input into a coherent steering force, creating a solid basis for organizational transformation. Finally, the management of IS implementation processes is a careful balancing act between initiating organizational change, and drawing upon IS as a change agent, without attempting to pre-specify and control this process. Accepting, and even drawing upon, this inevitable uncertainty might be the hardest lesson to learn. PMID- 11734383 TI - Integrating users' activity modeling in the design and assessment of hospital electronic patient records: the example of anesthesia. AB - As computers become more and more an aid in the management of medical information, some specialists, such as anesthesiologists, demand tuned applications to support their own activity. The development of these specific applications is based upon the user's requirements analysis, and functional and technical specifications. But some failures show that a better understanding of human factors of acceptance could improve the usability and utility of these tools. In this study, we demonstrated that when the management of medical information is closely intertwined with the physician's activity, it is necessary to perform a precise analysis of this activity in order to identify the cognitive and organizational constraints that affect the usability and acceptance of the tool. We focused our study on the pre-operative anesthetic consultation. After recording and analyzing 50 consultations, we were able to identify the key points to fulfill in order to meet users' acceptance. From this study, we propose some strong recommendations to handle the constraints imposed by the anesthesiologists' activity in their daily working environment. We applied this method to evaluate an electronic patient record (EPR) for the pre-anesthetic consultation. The results of this evaluation validate our hypotheses and the importance of the activity constraints. In conclusion, human factors, and particularly those linked with the activity of healthcare professionals, have to be carefully studied before any development and installation of an EPR into a specialty domain. PMID- 11734384 TI - Development and implementation of an EPR: how to encourage the user. AB - This paper reports on the role users played in the design and development of an electronic patient record. Two key users participated in the project team. All future users received questionnaires and a selection of them was interviewed. Before starting the development of the EPR, the attitude of users towards electronic record keeping, their satisfaction with the paper clinical records, their knowledge of computers, and their needs and expectations of computer applications in health care were measured by means of a questionnaire. The results of the questionnaire were supplemented with in-depth interviews. Users had a neutral attitude towards electronic record keeping. They were more positive about data entry of the paper records than data retrieval. During the development phase, but prior to the implementation of the EPR, a second questionnaire measured satisfaction with the paper records. Satisfaction appeared to be related to self-rated computer experience. Inexperienced computer users tended to be more positive about the paper records. In general, respondents did not have many expectations about electronic record keeping. A second series of interviews zoomed in on the expectations users had. Except for more concise reporting no beneficial effects of electronic record keeping were expected. PMID- 11734385 TI - Nursing process documentation systems in clinical routine--prerequisites and experiences. AB - Documentation of the nursing process is an important, but often neglected part of clinical documentation. Paper-based systems have been introduced to support nursing process documentation. Frequently, however, problems, such as low quality and high writing efforts, are reported. However, it is still unclear if computer based documentation systems can reduce these problems. At the Heidelberg University Medical Center, computer-based nursing process documentation projects began in 1998. A computer-based nursing documentation system has now been successfully introduced on four wards of three different departments, supporting all six phases of the nursing process. The introduction of the new documentation system was accompanied by systematic evaluations of prerequisites and consequences. In this paper, we present preliminary results of this evaluation, focusing on prerequisites of computer-based nursing process documentation. We will discuss in detail the creation and use of predefined nursing care plans as one important prerequisite for computer-based nursing documentation. We will also focus on acceptance issues and on organizational and technical issues. PMID- 11734386 TI - Electronic patient records and innovation in health care services. AB - The approach of today's EPR seems so narrowly focused on automation of the existing paper-based records by means of information technology that it becomes obvious to raise the question: 'Will these automation efforts become an impediment to innovation in products and services in health care?'. This paper discusses how objectives like improvements and innovations in products and services in health care are met by means of information technology (IT), and it argues that a shift of focus from technological innovation to innovation in products and services is necessary in order to obtain maximum benefit from IT. PMID- 11734387 TI - Medical knowledge reengineering--converting major portions of the UMLS into a terminological knowledge base. AB - We describe a semi-automatic knowledge engineering approach for converting the human anatomy and pathology portion of the UMLS metathesaurus into a terminological knowledge base. Particular attention is paid to the proper representation of part-whole hierarchies, which complement taxonomic ones as a major hierarchy-forming principle for anatomical knowledge. Our approach consists of four steps. First, concept definitions are automatically generated from the metathesaurus, with LOOM as the target language. Second, integrity checking of the emerging taxonomic and partonomic hierarchies is automatically carried out by the terminological classifier. Third, terminological cycles and inconsistencies are manually eliminated and, in the last step, the knowledge base built this way is incrementally refined by a medical expert. Our experiments were run on a terminological knowledge base which is composed of 164,000 concepts and 76,000 relations. Empirical evidence for the lack of logical consistency, adequacy and improper granularity of the UMLS knowledge source is given, and finally, assessments of what kind of efforts are needed to render the formal target representation structures complete and empirically adequate. PMID- 11734388 TI - Standardized terminological services enabling semantic interoperability between distributed and heterogeneous systems. AB - The interconnection of heterogeneous computer applications in medicine raises the issue of semantic interoperability, going beyond traditional approaches of terminological standardization in basically three aspects. First, the variety of medical vocabularies that currently coexist in different domains is a major barrier for the integration of autonomously developed applications. Fortunately, with the Unified Medical Language System (UMLS) there are machine-readable terminological sources that cover and integrate most of the existing medical vocabularies. Second, the exchanged data need to be processed by a machine for different purposes like patient data integration, access to literature and knowledge bases as well as clinical audit and research. Medical vocabularies provided as passive dictionaries are no longer sufficient. Software system developers should take advantage of terminological services for refining user queries, for mapping the user's terms to appropriate medical vocabularies etc. Third, the services should be accessible uniformly and transparently. In the CORBAmed initiative a proposal for a standardized interface for querying and accessing computerized medical terminology resources was created. Based on the mentioned principles the MUSTANG system (Medical UMLS based Terminology Server for Authoring, Navigating and Guiding the Retrieval to Heterogeneous Knowledge Sources) has been developed. It is implemented on a Windows NT platform using the ORACLE database management and development software. The terminological services are accessible via multiple interfaces. The MUSTANG-System and the experiences with using terminological services in practice are described. Opposed to other levels of standardization like syntactical message standards there is much more a hesitation in the use of standardized terminology. PMID- 11734389 TI - A MeSH based intelligent search intermediary for Consumer Health Information Systems. AB - GIN Austria (Gesundheitsinformationsnetz Austria) offers patients and consumers reliable medical knowledge about diseases, wellness and disease management in an easy understandable way and enables them to quick and incessant access to informations about the Austrian health system and Austrian health organizations. To achieve full customer (patients, citizens) satisfaction to find relevant information we propose a concept of a vector-model oriented IR-Tool based on the controlled vocabulary of the MeSH Thesaurus (german version). By this approach users who are often not used to scientific terms and expressions are supported to build up their own query with MeSH Main Headings. In a second step broader and narrower Main Headings are added to the query vector by the system. For this calculation an adapted version of the Floyd-Warshall algorithm for directed, azyclic graphs is used. The tool is part of the GIN Search Modul, which will ease gathering health information from different heterogenous internet datasources. PMID- 11734390 TI - HONselect: a multilingual and intelligent search tool integrating heterogeneous web resources. PMID- 11734391 TI - Conceptual alignment of electronic health record data with guideline and workflow knowledge. AB - Even though computerized practice guidelines and workflow management (WfM) are proven effective techniques to improving quality of care and reducing costs, they are not widely deployed today. One reason for this is the impedance mismatch between guideline systems and electronic health record (EHR) systems. This paper presents the Unified Service Action Model (USAM) that has been developed for the HL7 Reference Information Model (RIM) and that conceptually integrates guidelines and WfM in the EHR. We argue that the information items recorded in the EHR are logically similar to elements of guideline and WfM definitions. Therefore, the USAM suggests that guidelines and EHR reuse the same information structures. This reuse is possible through a technique borrowed from natural language grammar and modal logic. The conceptual alignment of guidelines, WfM and the EHR could facilitate the sharing and deployment of guidelines in routine health care. PMID- 11734392 TI - Computerising a guideline for the management of diabetes. AB - This paper reports an experience of computerising a clinical guideline for the management of non-insulin-dependent diabetes mellitus (NIDDM). The guideline, designed by the European NIDDM Policy Group is being used in a National Programme for Diabetes supported by the Portuguese Ministry of Health, who is keen to supporting its widespread use by general practitioners, namely in computerised form. The paper presents the main characteristics of the prototype that was implemented within the European project Prestige, and was developed according to the Prestige Protocol Model. The model is briefly described, together with the generic architecture that supports it. Then the main design decisions of the prototype are explained, regarding the modelling of a general practitioner workflow during a typical consultation and the user interface, two key issues for obtaining acceptance from the users. The limitations of the system are discussed and a number of directions are outlined in order to circumvent such limitations, and broaden the scope of applicability of the system. PMID- 11734393 TI - Design and implementation of a framework to support the development of clinical guidelines. AB - This paper describes and discusses a framework that facilitates the development of clinical guideline application tasks. The framework, named GASTON covers all stages in the guideline development process, ranging from the definition of models that represent guidelines to the implementation of run-time systems that provide decision support, based on the guidelines that were developed during the earlier stages. The GASTON framework consists of (1) a newly developed guideline representation formalism that uses the concepts of primitives, problem-solving methods (PSMs) and ontologies to represent the guidelines of various complexity and granularity and different application domains, (2) a guideline authoring environment that enables guideline authors to define the guidelines, based on the newly developed guideline representation formalism and (3) a guideline execution environment that translates defined guidelines into a more efficient symbol level representation, which can be read in and processed by an execution time engine. The paper describes a number of design criteria that were formulated regarding the aspects of guideline representation, guideline authoring and guideline execution and explains the framework by example in terms of the four stages that were identified in the guideline development process and the tools that were developed to support each stage. It also shows examples of systems that were developed by means of the GASTON framework. PMID- 11734394 TI - DiasNet--a diabetes advisory system for communication and education via the internet. AB - Intensive diabetes treatment can lead to a substantial reduction of the rate of the complications associated with diabetes. However, a number of patients may have poor control despite specialist care, and this along with devolution of care to non-specialists suggests that alternative interventions should be developed. The present paper describes an Internet based system where more emphasis is put on patient empowerment, the keywords being education and communication. The DiasNet system is based on a well documented decision support system, Dias, designed for use by clinicians. The scope of DiasNet has been widened from being used by clinicians to give advice on insulin dose, to also being used by patients as a tool for education and communication. Patients can experiment with their own data, adjusting insulin doses or meal sizes. In this way different therapeutic and dietary alternatives can be tried out, allowing the patient to gain experience in achieving glycaemic control. DiasNet is implemented in JAVA according to the client/server principle, enabling a new way of communication between patient and clinician: in case of any problems, the patient simply phones the clinician, who immediately, using his or her office PC, can take a look at the data the patient has entered. PMID- 11734395 TI - Decision support for infectious diseases--a working prototype. AB - This paper presents a decision support system for nosocomial infections and its integration in the large HIS of the University Hospital of Giessen. The system comprises five different engines and a data dictionary. It is designed to detect hospital acquired infections even in a situation where only a restricted amount of clinical data is available (the data is split up in different information systems). Furthermore the model prevents time consuming manual data entry. The five engines split the main task into: (1) a preselection, which sorts out patients who definitely do not have a nosocomial infection; (2) a rule based reasoning process which detects patients likely to have such an infection; (3) an alarm process which is responsible for the presentation of the alert; (4) an explanation process to follow up the reasoning; and (5) statistic tools to answer specific hygienic questions. A data dictionary supplies the controlled vocabulary, which is required to understand data structures used in the different clinical subsystems and may those with each other. PMID- 11734396 TI - Validation of a knowledge based reminder system for diagnostic test ordering in general practice. AB - We describe the validation of a real-time automated reminder system that assists General Practitioners (GP) in appropriate test ordering. We compared the comments of human experts with the comments of the reminder system using a retrospective random selection of 253 request forms. A panel of three expert physicians judged the requested tests independently based on their interpretations of the practice guidelines. The majority assessment of the physicians was compared with the assessment of the reminder system. In case the system's output differed from the majority assessment the written practice guidelines were consulted. On average 1.75 reminders were produced per form. In total 32 of the 442 given reminders (7%) were given incorrectly. The amount of information and the level of detail (the specificity of the terms) in which the GP describes the patients' medical status are crucial for the reminder system to react correctly. PMID- 11734397 TI - Cased-Based Reasoning for medical knowledge-based systems. AB - In this paper we present the results of the MIE/GMDS-2000 Workshop 'Case-Based Reasoning for Medical Knowledge-based Systems'. While in many domains Cased-Based Reasoning (CBR) has become a successful technique for knowledge-based systems, in the medical field attempts to apply the complete CBR cycle are rather exceptional. Some systems have recently been developed, which on the one hand use only parts of the CBR method, mainly the retrieval, and on the other hand enrich the method by a generalisation step to fill the knowledge gap between the specificity of single cases and general rules. And some systems rely on integrating CBR and other problem solving methodologies. In this paper we discuss the appropriateness of CBR for medical knowledge-based systems, point out problems, limitations and possible ways to cope with them. PMID- 11734398 TI - Towards integration of clinical decision support in commercial hospital information systems using distributed, reusable software and knowledge components. AB - PROBLEM: Clinicians' acceptance of clinical decision support depends on its workflow-oriented, context-sensitive accessibility and availability at the point of care, integrated into the Electronic Patient Record (EPR). Commercially available Hospital Information Systems (HIS) often focus on administrative tasks and mostly do not provide additional knowledge based functionality. Their traditionally monolithic and closed software architecture encumbers integration of and interaction with external software modules. Our aim was to develop methods and interfaces to integrate knowledge sources into two different commercial hospital information systems to provide the best decision support possible within the context of available patient data. METHODS: An existing, proven standalone scoring system for acute abdominal pain was supplemented by a communication interface. In both HIS we defined data entry forms and developed individual and reusable mechanisms for data exchange with external software modules. We designed an additional knowledge support frontend which controls data exchange between HIS and the knowledge modules. Finally, we added guidelines and algorithms to the knowledge library. RESULTS: Despite some major drawbacks which resulted mainly from the HIS' closed software architectures we showed exemplary, how external knowledge support can be integrated almost seamlessly into different commercial HIS. This paper describes the prototypical design and current implementation and discusses our experiences. PMID- 11734399 TI - XML for electronic clinical communications in Scotland. AB - This paper describes the use of extensible markup language (XML) for electronic clinical communications in Scotland. It gives the results of the first UK project to send and receive discharge letters using XML. This pilot had the advantage of transferring readable clinical information between computer systems and giving the receiving GP choice of decoding and saving clinical and administrative data into his practice system. A number of existing clinical systems and a variety of written documents were studied. Existing trial standards, including the NHS EDIFACT, HL7v2 and CEN standard ENV 13606 were taken into consideration. The project worked with the receiving GP system supplier, GPASS, and the NHS Scotland Information and Statistics Division in defining the document type definition (DTD) for the pilot DTD. GPASS undertook production of a module for their system to parse the XML messages into the patient record on the GP system. The system allows the GP to read and print the original document and modify the content prior to storing in the practice system. SMTP e-mail messages were used for the pilot. Resulting messages were parsed into the GP system and validated by a variety of GP users. Currently there is a large scale program to develop XML Schema and web browser based communications of referrals, discharge and laboratory orders and reports. PMID- 11734400 TI - Medical support system for continuation of care based on XML web technology. AB - In this paper, the implementation of an Internet-based telematic service for medical support is presented, which was developed and operated in pilot form within the INTRANET HEALTH CLINIC project--a 2-year project supported by the European Commission under the Health Telematics Programme. The aim of the application is to offer high quality care to users of health services over inexpensive communication pathways, using Internet-based, interactive communication tools, like remote access to medical records and transmission of multimedia information. The XML technology was employed to achieve customised views on patient data, according to the access rights of different user profiles. Strict security and access control policy were implemented to ensure secure transmission of medical data through the Internet. The system was designed to collaborate with existing clinical patient record systems and to be adjustable to different medical applications. Current implementations include the fields of Oncology, Lupus Erythrematosis, Obstetrics and Chronic Obstructive Pulmonary disease. The results of the pilot operation with oncological patients in Greece were encouraging, so that the refining of the system and its expansion to a large number of patients is already in progress. PMID- 11734401 TI - Securing interoperability between chip card based medical information systems and health networks. AB - Health information systems supporting shared care are going to be distributed and interoperable. Dealing with sensitive personal medical information, such information systems have to provide appropriate security services, allowing only authorised users restricted access rights to the patients' data according to the 'need to know' principle. Especially in healthcare, chip card based information systems occur in the shape of patient data cards providing informational self determination and mobility of the users as well as quality, integrity, accountability, and availability of the data stored on the card, thus improving the shared care of patients. The DIABCARD project aims at the implementation and evaluation of a chip card based medical information system (CCMIS) for facilitating communication and co-operation between health professionals in different organisations or departments caring the same patient with diabetes as an example. In co-operation with the EC-funded TrustHealth(2) project, communication and application security services needed are provided like strong authentication as well as the derived services such as authorisation, access control, accountability, confidentiality, etc. The solution is based on Health Professional Cards and Trusted Third Party services. In addition to the secure handling of the patient's chip card and data in DIABCARD workstations, the secure communication between these workstations and related departmental systems has been implemented. Based on the results of this feasibility study, an enhanced security services specification for the DIABCARD example of a CCMIS is provided which will be implemented in the framework of a health network being established in the German federal state Bavaria. Beside the preferred solution of a combination of Patient Identification Card and Patient Data Card, lower level alternatives using card-verifiable certificates are explained in some details. Finally, a few legal issues, future trends like the XML standard set and their implications for the solution presented as well as for distributed health information systems in general are shortly discussed. PMID- 11734402 TI - Secure medical digital libraries. AB - In this paper, a secure medical digital library is presented. It is based on the CORBA specifications for distributed systems. The described approach relies on a three-tier architecture. Interaction between the medical digital library and its users is achieved through a Web server. The choice of employing Web technology for the dissemination of medical data has many advantages compared to older approaches, but also poses extra requirements that need to be fulfilled. Thus, special attention is paid to the distinguished nature of such medical data, whose integrity and confidentiality should be preserved at all costs. This is achieved through the employment of Trusted Third Parties (TTP) technology for the support of the required security services. Additionally, the proposed digital library employs smartcards for the management of the various security tokens that are used from the above services. PMID- 11734403 TI - A JAVA-based DICOM server with integration of clinical findings and DICOM-conform data encryption. AB - The transfer of large amounts of medical data within heterogeneous hard and software infrastructures and the exploitation of distributed resources require a fast, secure, and platform-independent data exchange. To avoid costly vendor specific solutions, a DICOM server was implemented in JAVA. Data access was enabled via internet browser technology. Relevant patient and image acquisition information was extracted from the DICOM images and stored into a relational database. Patient information such as radiological findings were transferred from the Radiological Information System into the database. Image data were accessed either by a fast preview tool or using a JAVA-based DICOM viewer. Since data security mechanisms are not yet part of the DICOM standard, a DICOM-conform encryption of sensitive patient data was implemented. The method allowed a dynamic selection of the data to be encrypted. Integrating this module into the image server enabled the fast and secure transfer of image data across insecure networks as well as long-term storage on CD-Recordables. PMID- 11734404 TI - Atlas-based segmentation of bone structures to support the virtual planning of hip operations. AB - Two 3-D digitised atlases of a female and a male pelvis were generated to support the virtual 3-D planning of hip operations. The anatomical atlases were designed to replace the interactive, time-consuming pre-processing steps for the virtual operation planning. Each atlas consists of a labelled reference CT data set and a set of anatomical point landmarks. The paper presents methods for the automatic transfer of these anatomical labels to an individual patient data set. The labelled patient data are used to generate 3-D models of the patient's bone structures. Besides the anatomical labelling, the determination of measures, like angles, distances or sizes of contact areas, is important for the planning of hip operations. Thus, algorithms for the automatic computation of orthopaedic parameters were implemented. A first evaluation of the presented atlas-based segmentation method shows a correct labelling of 98.5% of the bony voxels. PMID- 11734405 TI - Reality and perspectives in teleradiology: a personal view based on personal experiences. PMID- 11734406 TI - Simulating tumour removal in neurosurgery. AB - In this article the software system ROBO-SIM is described. ROBO-SIM is a planning and simulation tool for minimally invasive neurosurgery. Different to the most other simulation tools, ROBO-SIM is able to use actual patient's datasets for simulation. Same as in real neurosurgery a planning step, which provides more functionality as up-to-date planning systems on the market, is performed before undergoing the simulated operation. The planning steps include the definition of the trepanation point for entry into the skull and the target point within the depth of the brain, checking the surgical track and doing virtual trepanations (virtual craniotomy). For use with an intra-operative active manipulator, which is guided by the surgeon during real surgery (robotic surgery), go- and non-go areas can be defined. During operation, the robot restricts the surgeon from leaving these go-areas. After planning, an additional simulation system, which is understood as an extension to the planning step, is used to simulate whole surgical interventions directly on the patient's anatomy basing on the planning data and by using the same instruments as for the real intervention. First tests with ROBO-SIM are performed on a phantom developed for this purpose and on actual patient's datasets with ventricular tumours. PMID- 11734407 TI - Ultrasound and angio image compression by cosine and wavelet transforms. AB - The investigation results for improving lossy compression techniques for ultrasound and angio images are presented. The goal was to determine where the compression process could be improved for the medical application, and to make efforts to improve it. It is proved that the wavelet transform outperforms the discrete cosine transform applied to ultrasound and angio images. A lot of wavelet classes were tried for choosing the best one suited for corresponding image classes, which were characterised by a content complexity criterion. The analysis of international image compression standards was carried out. Special attention was paid to an algorithmical and high level service structure of a new still image compression standard JPEG2000. Its open architecture enables including some wavelet classes which we would like to suggest for medical images. A set of recommendations for acceptable compression ratio for different medical image modalities was developed. It was carried out on the base of compression study performed by the group of angiologists and cardiologists. PMID- 11734408 TI - Attunement, calibration, and exploration in fast haptic perceptual learning. AB - Often, a relatively small number of trials suffices to enhance one's task specific perceptual capability. In the present experiment, fast perceptual learning was investigated with respect to the perception of the heights or widths of wielded nonvisible rectangular objects. In that haptic perceptual task, inertial differences (mass and moments of inertia) are the basis for perceived size differences. The authors hypothesized that rapid improvement might occur in attunement (attending to the task-relevant inertial variable), calibration (scaling spatial extent to the task-relevant inertial variable), and exploratory behavior (wielding so as to differentiate the task-relevant inertial variable). Twenty-four students performed 25 trials with a set of practice objects; those trials were followed and preceded by 18 trials with a set of test objects. Practice, with knowledge of results (KR), improved both attunement, as measured by regression of perceived spatial extent on the inertial variables, and calibration, as measured by constant and variable error. Of the preceding measures, only variable error improved with practice in the absence of KR. In both KR conditions, however, exploratory behavior decreased in duration and complexity, as measured by recurrence quantification analysis. The present results suggest that the mechanisms involved in fast perceptual learning are more varied and complex than are those encompassed by current accounts. PMID- 11734409 TI - Sinusoidal visuomotor tracking: intermittent servo-control or coupled oscillations? AB - In visuomotor tasks that involve accuracy demands, small directional changes in the trajectories have been taken as evidence of feedback-based error corrections. In the present study variability, or intermittency, in visuomanual tracking of sinusoidal targets was investigated. Two lines of analyses were pursued: First, the hypothesis that humans fundamentally act as intermittent servo-controllers was re-examined, probing the question of whether discontinuities in the movement trajectory directly imply intermittent control. Second, an alternative hypothesis was evaluated: that rhythmic tracking movements are generated by entrainment between the oscillations of the target and the actor, such that intermittency expresses the degree of stability. In 2 experiments, participants (N = 6 in each experiment) swung 1 of 2 different hand-held pendulums, tracking a rhythmic target that oscillated at different frequencies with a constant amplitude. In 1 line of analyses, the authors tested the intermittency hypothesis by using the typical kinematic error measures and spectral analysis. In a 2nd line, they examined relative phase and its variability, following analyses of rhythmic interlimb coordination. The results showed that visually guided corrective processes play a role, especially for slow movements. Intermittency, assessed as frequency and power components of the movement trajectory, was found to change as a function of both target frequency and the manipulandum's inertia. Support for entrainment was found in conditions in which task frequency was identical to or higher than the effector's eigenfrequency. The results suggest that it is the symmetry between task and effector that determines which behavioral regime is dominant. PMID- 11734410 TI - Dynamics of pushing. AB - A standing individual can use several strategies for modulating pushing force magnitude. Using a static model, researchers have shown that the efficacy of those strategies varies considerably. In the present article, the authors propose a human motor control dynamic model for analyzing transients that occur when an individual is asked to modulate force magnitude. According to the model, the impedances of both the upper and the lower limbs influence the time course of force variations and foot placement has a profound effect on pushing force dynamics. With a feet-together posture, the center of pressure has a limited range of motion and changes in force may be preceded by initial changes in the opposite direction; that is, to decrease force, an individual must first increase force. When the feet are placed apart, individuals can move the center of pressure over a much larger range, thereby modulating pushing force magnitude, without reversing behavior, over a larger range of force magnitudes. Therefore, the best way to control pushing force at the hand may be by using the foot. PMID- 11734411 TI - Exploration and selection of intralimb coordination patterns in 3-month-old infants. AB - Through the exploration of their own capacities and the selection of adaptive responses, infants learn new motor solutions. Using a conjugate reinforcement mobile procedure, previous researchers have repeatedly shown that infants increase their leg kick frequency to control a mobile that is connected to their ankles. That traditional experimental design allows multiple motor solutions to the task and therefore provides limited information about the infants' capacity to explore and select specific motor solutions. The author designed a new experimental procedure to study infants' capacity to discover and adopt specific motor solutions. The new, constraining mobile reinforcement procedure requires a specific motor response and therefore the development of a more finely tuned perception-action map than has previously been experimentally demonstrated. To gain reinforcement from the mobile, infants had to produce a coordinated hip and knee extension within the same leg. The results from the 13 infant participants showed that they were capable of increasing their frequency of coordinated movements to make the mobile move. Those results suggest that infants at the age of 89-106 days are sensitive to intralimb coordination task requirements and are capable of mapping their own limb dynamics to the environmental information. PMID- 11734412 TI - Mental and motor switching in Parkinson's disease. AB - Switching difficulties in Parkinson's disease (PD) are expressed in both mental and motor tasks. The authors of the present study investigated whether those deficits coexist in the same patient and are positively correlated. They tested 8 nondemented PD patients and 6 age-matched control participants by using the modified Wisconsin Card Sorting Test and a motor switching paradigm that is based on the task of reaching toward visual targets, the location of which could unexpectedly be altered within the reaction time. In both mental and motor tasks, patients performed significantly worse than controls. There were no significant correlations between the two types of pathology in individual patients. Mental and motor switching deteriorate in PD patients, but the deficits are not necessarily of parallel severity. PMID- 11734413 TI - A modeling study of potential sources of curvature in human reaching movements. AB - The authors of this article suggest that the slight but consistent posture dependent curvature of the spatial paths in the kinematic transformation between intrinsic and extrinsic coordinates may result in a systematic curvature of movements initially planned as straight-line trajectories toward the target. A kinematic planning model is presented that takes into account the anisotropy of the intrinsic and extrinsic transformation and tends to avoid movements that require excessive joint rotations by introducing slight deviations from a straight-line trajectory. Preliminary simulations showed reasonably good agreement with experimental data, especially considering that the current model is strictly based on kinematics. A quantitative analysis showed that the strategy used in the model achieves a favorable compromise between straight-line movements and angular joint changes: By slightly increasing the spatial length of the movement (i.e., by introducing curvature), an individual can greatly reduce the total amount of joint rotation required to produce the movement. PMID- 11734414 TI - Calibration and alignment are separable: evidence from prism adaptation. AB - In 2 prism adaptation experiments, the authors investigated the effects of limb starting position visibility (visible or not visible) and visual feedback availability (early or late in target pointing movements). Thirty-two students participated in Experiment 1 and 24 students participated in Experiment 2. Independent of visual feedback availability, constant error was larger and variable error was smaller for target pointing when limb starting position was visible during prism exposure. Independent of limb starting position visibility, aftereffects of prism exposure were determined by visual feedback availability. Those results support the hypothesis that calibration is determined by limb starting position visibility, whereas alignment is determined separately by visual feedback availability. PMID- 11734415 TI - Fallingwater and emphysema. PMID- 11734416 TI - The ACCESS study: characterization of sarcoidosis in the United States. PMID- 11734417 TI - Bronchopulmonary dysplasia: "a vascular hypothesis". PMID- 11734418 TI - Airway infection accelerates decline of lung function in chronic obstructive pulmonary disease. PMID- 11734419 TI - Airway infection does not accelerate decline in lung function in chronic obstructive pulmonary disease. PMID- 11734422 TI - When the chest X-ray does not tell the whole story: a tale of miners, selection bias, and the healthy worker effect. PMID- 11734423 TI - Practical management of diabetes in critically ill patients. PMID- 11734424 TI - HMGB1 as a late mediator of lethal systemic inflammation. PMID- 11734425 TI - Tuberculosis, lung infections, and interstitial lung disease in AJRCCM 2000. PMID- 11734426 TI - Chronic obstructive pulmonary disease, pollution, pulmonary vascular disease, transplantation, pleural disease, and lung cancer in AJRCCM 2000. PMID- 11734427 TI - Longitudinal changes in physiological, radiological, and health status measurements in alpha(1)-antitrypsin deficiency and factors associated with decline. AB - The FEV(1) declines rapidly in alpha(1)-antitrypsin deficiency (alpha(1)-ATD) but less is known about other measures of disease severity and the factors, other than smoking, that are associated with progression of emphysema. The natural history of alpha(1)-ATD was studied prospectively in 43 patients with the PiZ phenotype and emphysema at a single center over 2 yr. The mean +/- SE change in FEV(1) was -67 +/- 14 ml/yr, accompanied by a reduction in transfer factor (mean change in diffusing capacity of the lung for CO [DL(CO)] -1.07 +/- 0.21 ml/min/mm Hg/yr; p < 0.001) and lung density in the upper zones as assessed by quantitative high-resolution computed tomography (HRCT) (mean change in voxel index 2.8 +/- 0.6%/yr; p < 0.001). The decline in FEV(1) related to baseline FEV(1) (r = -0.56, p < 0.001), bronchodilator reversibility (r = 0.52, p < 0.001), and (for patients with FEV(1) > 35% predicted) exacerbation frequency (r = -0.38, p = 0.02). There was also a decline in the St. George's Respiratory Questionnaire (SGRQ) Activity score (mean change -4.3 +/- 1.2 units/yr, p < 0.001) that correlated with FEV(1) decline (r = 0.45, p = 0.002). Progression of emphysema in alpha(1)-ATD is dependent on baseline physiology and exacerbation frequency and may be detected by several different measurements of which HRCT density mask analysis and DL(CO) appear most sensitive. PMID- 11734428 TI - Airway response to inhaled hypertonic saline in patients with moderate to severe chronic obstructive pulmonary disease. AB - The present study aimed at delineating the mechanisms underlying the adverse response to hypertonic saline inhalation in patients with chronic obstructive pulmonary disease (COPD). Twenty patients (age, 48-70 yr; FEV(1), 29-58 %pred) inhaled, on two different days in randomized order, 200 microg salbutamol from an MDI and 20 min later either 0.9% or 3% saline from an ultrasonic nebulizer for a maximum of four consecutive 5-min periods. Forced expiratory (FEV(1)) and inspiratory (FIV(1)) volumes, inspiratory capacity (IC), intrathoracic gas volume (ITGV), and specific airway resistance (SRaw) were measured. Significant changes occurred in FEV(1), FIV(1), IC, ITGV, and SRaw with both concentrations (p < 0.05, each) and effects were stronger with 3% as compared with 0.9% saline (p < 0.05, each). The increase in dyspnea was associated with the changes in FIV(1), FEV(1), IC, and ITGV, in contrast to its decrease during bronchodilation, where only FIV(1) was important. Sputum analysis showed elevated concentrations of histamine after 3% as compared with 0.9% saline. These data indicate that the adverse lung function response to hypertonic saline is common in patients with moderate to severe COPD, involves both bronchoconstriction and lung hyperinflation, and could be mediated, at least partially, through activation of mast cells. PMID- 11734429 TI - Rhinovirus-induced airway inflammation in asthma: effect of treatment with inhaled corticosteroids before and during experimental infection. AB - Asthma exacerbations are frequently linked to rhinovirus infections. However, the associated inflammatory pathways are poorly understood, and treatment of exacerbations is often unsatisfactory. In the present study we investigated whether antiinflammatory treatment with inhaled corticosteroids prevents any rhinovirus-induced worsening of lower airway inflammation. To that end, we selected 25 atopic patients with mild asthma who underwent experimental rhinovirus 16 (RV16) infection, while receiving double-blind, placebo-controlled treatment with the inhaled corticosteroid budesonide (800 microg twice a day) throughout the study period, starting 2 wk before infection. We assessed inflammatory cell numbers in the bronchial mucosa as obtained by bronchial biopsies 2 d before and 6 d after RV16 infection, and analyzed those in relation to cold symptoms, changes in blood leukocyte counts, airway obstruction, and airway hyperresponsiveness. RV16 colds induced an increase in CD3(+) cells in the lamina propria (p = 0.03) and tended to decrease the numbers of epithelial eosinophils (p = 0.06) in both groups analyzed as a whole. The T cell accumulation was positively associated with cold symptoms. Budesonide pretreatment improved airway hyperresponsiveness (p = 0.02) and eosinophilic airways inflammation (p = 0.04). Yet it did not significantly affect the RV16 associated changes in the numbers of any of the inflammatory cell types. We conclude that RV16 infection by itself induces only subtle worsening of airway inflammation in asthma, which is not improved (or worsened) by inhaled corticosteroids. The latter finding is in keeping with the limited protection of inhaled corticosteroids against acute asthma exacerbations. PMID- 11734430 TI - Dietary antioxidants and asthma in adults: population-based case-control study. AB - A protective role for dietary antioxidants in asthma has been proposed. However, epidemiological evidence to implicate antioxidant vitamins is weak, and data on the role of flavonoid-rich foods and antioxidant trace elements are lacking. We carried out a population-based case-control study in South London, UK, to investigate whether asthma is less common and less severe in adults who consume more dietary antioxidants. Participants were aged 16-50 yr and registered with 40 general practices. Asthma was defined by positive responses to a standard screening questionnaire in 1996, and complete information about usual diet was obtained by food frequency questionnaire from 607 cases and 864 controls in 1997. After controlling for potential confounding factors and total energy intake, apple consumption was negatively associated with asthma (odds ratio [OR] per increase in frequency group 0.89 [95% confidence interval [CI]: 0.82 to 0.97]; p = 0.006). Intake of selenium was also negatively associated with asthma (OR per quintile increase 0.84 [0.75 to 0.94]; p = 0.002). Red wine intake was negatively associated with asthma severity. The associations between apple and red wine consumption and asthma may indicate a protective effect of flavonoids. The findings for dietary selenium could have implications for health policy in Britain where intake has been declining. PMID- 11734431 TI - Does living on a farm during childhood protect against asthma, allergic rhinitis, and atopy in adulthood? AB - Recent studies of children suggest that factors encountered in a farm environment might protect against the development of allergy. However, it remains uncertain whether living on a farm in childhood is associated with a decreased risk of atopic diseases in adulthood. We analyzed data from 6,251 randomly selected adults 20 to 44 yr of age participating in the European Community Respiratory Health Survey (ECRHS). Subjects answered a detailed questionnaire and underwent specific IgE measurements to five allergens. After adjustment for potential confounders, including pet exposure in childhood, number of siblings, severe respiratory infection in childhood, and parental history of allergy, living on a farm in childhood was associated with a reduced risk of atopic sensitization in adulthood (OR = 0.76, CI 95% = 0.60-0.97). Compared with other adults, those who had lived on a farm as a child were less frequently sensitized to cat (OR = 0.63, CI 95% = 0.41-0.96) and to Timothy grass (OR = 0.68, CI 95% = 0.50-0.94), and were at lower risk of having nasal symptoms in the presence of pollen (OR = 0.80, CI 95% = 0.64-1.02). The protective effect of farming environment in childhood observed in this population-based sample of young adults provides evidence in favor of the hypothesis that environmental factors encountered in childhood may have a lifelong protective effect against the development of allergy. PMID- 11734432 TI - Body mass index and asthma in adults in families of subjects with asthma in Anqing, China. AB - We conducted a cross-sectional study of 7,109 adults from families of subjects with asthma in the province of Anhui, China. Asthma was defined either as a combination of physician-diagnosed asthma, airway responsiveness to methacholine at < or = 25 mg/ml and two or more respiratory symptoms or asthma attacks ("asthma"); or as a combination of airway responsiveness to methacholine at < or = 8 mg/ml and two or more respiratory symptoms or asthma attacks ("symptomatic airway hyperresponsiveness [AHR]"). After adjusting for intensity of cigarette smoking and other variables, both extremes of the body mass index (BMI) distribution were associated with symptomatic AHR in men and women (p < 0.01). In the multivariate analysis, both under- and overweight were associated with asthma in women, and underweight was associated with asthma in men. Among men, those with BMIs of 16 and 30 kg/m(2) had 2.5 and 2.3 times higher odds of symptomatic AHR, respectively, than those whose BMI was 21 kg/m(2) (95% CI for OR(16 vs. 21 kg/m)(2) = 1.4 to 3.8; 95% CI for OR(30) (vs.) (21) (kg/m)(2) = 1.2 to 5.0). Among women, those with BMIs of 16 and 30 kg/m(2) had 2.0 and 2.3 times higher odds of symptomatic AHR than those whose BMI was 21 kg/m(2) (95% CI for OR(16) (vs.) (21) (kg/m)(2) = 1.3 to 3.1; 95% CI(30) (vs.) (21) (kg/m)(2) = 1.2 to 4.5). Among adults in families of subjects with asthma living in rural China, both underweight and overweight are associated with an increased risk of asthma. PMID- 11734433 TI - Complement factors C3a and C5a are increased in bronchoalveolar lavage fluid after segmental allergen provocation in subjects with asthma. AB - Allergic asthma is thought to be the result of an inappropriate specific immune response against common environmental antigens. However, studies of animal asthma models have also linked the innate immune system, in particular complement factors C3a and C5, to murine airway hyperresponsiveness. Because the possible role of these anaphylatoxins in patients with asthma is not understood, we tested the hypothesis that C3a and C5a will increase in the bronchoalveolar lavage (BAL) fluid of patients with asthma after segmental allergen provocation. In a group of 15 subjects with mild asthma we found a significant upregulation of C3a and C5a 24 h after allergen challenge compared with baseline values (p < 0.01). In a control group of healthy volunteers the concentrations remained basically unchanged. Furthermore, we found a strong correlation between both anaphylatoxins and the number of eosinophils (p < 0.01) and, to a lesser degree, with the number of neutrophils (p < 0.05) in BAL fluid. These data suggest a contribution of anaphylatoxins C3a and C5a to the pathogenesis in asthma. However, the pathogenic role of these substances in relation to asthma remains to be elucidated, for example, by using anaphylatoxin receptor blockers as a possible new therapeutic principle. PMID- 11734434 TI - Alterations in exhaled gas profile during allergen-induced asthmatic response. AB - The source of exhaled carbon monoxide (CO) and the relationship to airway inflammation are not clear. If CO is produced by the inflamed airway, we hypothesized that inflammation induced by allergen challenge would increase exhaled CO of atopic asthmatics. Eight atopic asthmatics underwent whole lung allergen challenge. CO, nitric oxide (NO), oxygen, and carbon dioxide (CO(2)) were measured simultaneously in exhaled breath which was collected into Mylar balloons before (baseline), immediately after, and at subsequent times after allergen. NO was higher in asthmatics than control subjects at baseline, increased further in seven of the eight asthmatics after allergen, and was inversely correlated to specific conductance. In contrast, exhaled CO of asthmatics was not higher than that of control individuals at baseline, decreased immediately after allergen, and returned to baseline levels during the late asthmatic response. Thus, allergen-induced airway inflammation did not lead to increased exhaled CO in asthma. PMID- 11734435 TI - Estradiol decreases the acetylcholine-elicited airway reactivity in ovariectomized rats through an increase in epithelial acetylcholinesterase activity. AB - Estrogen replacement therapy (ERT) is frequently prescribed for postmenopausal women. Epidemiological data suggest that sex hormones may play a role in the expression of asthma, but the mechanism(s) whereby this influence is mediated remain(s) unclear. To better understand the role of physiologic doses of estrogens in airway function, we tested the hypothesis that 17beta-estradiol (E(2), 10 microg/kg per d for 21 d) given to oophorectomized female rats modifies airway responsiveness to cholinergic agonists, compared with oophorectomized rats given placebo. In vivo, the concentration of inhaled acetylcholine (ACh) required to double pulmonary resistance (EC(200)RL) in anesthetized spontaneously breathing tracheotomized rats was calculated as an index of airway responsiveness. E(2)-treated rats were less responsive to ACh than placebo treated rats (EC(200)RL, 9.40 +/- 1.48 vs. 1.52 +/- 0.85 mg. ml(-1), respectively). Ex vivo airway responsiveness was evaluated with the cumulative concentration-response curve (CCRC) of isolated tracheal segments. Compared with placebo, E(2) treatment significantly increased the EC(50) of ACh (p = 0.01) but did not alter the CCRC to carbachol. Removing the epithelium or treatment with physostigmine abolished the difference in EC(50) of ACh between the groups. Acetylcholinesterase (AChE) activity of homogenized whole trachea was 1.4-fold greater in the E(2)-treated group compared with placebo (p = 0.02), whereas no difference was found in homogenized epithelium-free trachea. We conclude that E(2) treatment decreases airway responsiveness to ACh in ovariectomized rats at least in part by increasing AChE activity dependent on the presence of the epithelium. PMID- 11734436 TI - Hyaluronic acid blocks porcine pancreatic elastase (PPE)-induced bronchoconstriction in sheep. AB - We previously showed that inhaled porcine pancreatic elastase (PPE) causes bronchoconstriction in sheep via a bradykinin-mediated mechanism. Hyaluronic acid (HA), in vitro, binds and inactivates airway tissue kallikrein (TK), the enzyme responsible for kinin generation. Therefore, we hypothesized that in vivo, HA should prevent PPE-induced bronchoconstriction by binding and inactivating TK. To test this, we measured pulmonary resistance (RL) in allergic sheep before and after inhalation of PPE alone (500 microg) and after pretreatment with either inhaled HA at 70 kD, designated low molecular weight (LMW)-HA or 200 kD, designated high molecular weight (HMW)-HA at different concentrations. Inhaled PPE increased RL 147 +/- 8% over baseline values and this effect was associated with a 111 +/- 28% increase in bronchoalveolar lavage fluid (BALF) TK activity. HA blocked the PPE-induced bronchoconstriction and the increase in BALF TK activity in a dose- dependent and molecular weight-dependent fashion. HA alone had no effect on RL. Instillation of PPE in the lung increased kinin concentrations in BALF, a result consistent with the PPE-induced increase in BALF TK activity. Our findings show that HA blocks PPE-induced bronchoconstriction in a dose-dependent and molecular weight-dependent fashion by a mechanism that may, in part, be related to inhibition of TK activity and the formation of kinins. PMID- 11734437 TI - Acute effects of particulate air pollution on respiratory admissions: results from APHEA 2 project. Air Pollution and Health: a European Approach. AB - The APHEA 2 project investigated short-term health effects of particles in eight European cities. In each city associations between particles with an aerodynamic diameter of less than 10 microm (PM(10)) and black smoke and daily counts of emergency hospital admissions for asthma (0-14 and 15-64 yr), chronic obstructive pulmonary disease (COPD), and all-respiratory disease (65+ yr) controlling for environmental factors and temporal patterns were investigated. Summary PM(10) effect estimates (percentage change in mean number of daily admissions per 10 microg/m(3) increase) were asthma (0-14 yr) 1.2% (95% CI: 0.2, 2.3), asthma (15 64 yr) 1.1% (0.3, 1.8), and COPD plus asthma and all-respiratory (65+ yr) 1.0% (0.4, 1.5) and 0.9% (0.6, 1.3). The combined estimates for Black Smoke tended to be smaller and less precisely estimated than for PM(10). Variability in the sizes of the PM(10) effect estimates between cities was also investigated. In the 65+ groups PM(10) estimates were positively associated with annual mean concentrations of ozone in the cities. For asthma admissions (0-14 yr) a number of city-specific factors, including smoking prevalence, explained some of their variability. This study confirms that particle concentrations in European cities are positively associated with increased numbers of admissions for respiratory diseases and that some of the variation in PM(10) effect estimates between cities can be explained by city characteristics. PMID- 11734438 TI - Induction of inflammatory response of mice exposed to diesel exhaust is modulated by CD4(+) and CD8(+) T cells. AB - Exposure to diesel exhaust (DE) increased airway inflammatory responses and airway responsiveness to allergen challenge. To clarify the roles of T cells in DE exposure-induced early inflammation, we studied the effect of CD4 and CD8 cells on the effect DE might have on allergic inflammation by using monoclonal antibody-mediated cellular depletion assays. In the bronchoalveolar lavage (BAL) fluid, the numbers of inflammatory cells from 3 mg/m(3) DE-exposed and ovalbumin (OVA)-immunized mice markedly increased. Depletion of CD4(+) cells resulted in reduced accumulation of inflammatory cells. DE exposure to OVA-immunized mice significantly increased interleukin (IL)-1 beta production but decreased IL-12 production. DE exposure significantly enhanced production of the macrophage inflammatory proteins (MIP)-1 alpha and MIP-2, but not monocyte chemoattractant protein (MCP)-1 and regulated upon activation normal T cells expressed and secreted (RANTES). Treatment with anti-CD4 and anti-CD8 mAbs abrogated the adverse effect of DE exposure. In CLN cells from OVA + DE-exposed mice, CD45R/B220-, CD3-, CD4-, and CD8-positive cells were significantly increased, but the OVA-stimulated cytokine production remained at the same levels with OVA immunized mice. These findings suggest that the induction of early inflammatory responses by DE exposure may initially be related to the modulated function of lymphocyte subpopulations. PMID- 11734439 TI - Chronic stridor caused by laryngomalacia in children: work of breathing and effects of noninvasive ventilatory assistance. AB - Breathing pattern, gas exchange, and respiratory effort were assessed in five awake children with chronic stridor caused by laryngomalacia during spontaneous breathing (SB) and noninvasive mechanical ventilation (NIMV). During SB, the youngest children were able to maintain normal gas exchange at the expense of an increased work of breathing as assessed by calculated diaphragmatic pressure-time product (PTPdi), whereas the opposite was observed in the older children. NIMV increased tidal volume, from 8.77 +/- 2.04 ml/kg during SB to 11.67 +/- 2.52 ml/kg during NIMV, p = 0.04, and decreased respiratory rate, from 24.4 +/- 5.6 breaths/ min during SB to 16.6 +/- 0.9 breaths/min during NIMV, p = 0.04. NIMV unloaded the respiratory muscles as reflected by the significant reduction in PTPdi, from a mean value of 541.0 +/- 196.6 cm H(2)O x s x min(-1) during SB to 214.8 +/- 116.0 cm H(2)O x s x min(-1) during NIMV, p = 0.04. Therefore, NIMV successfully relieves the additional load imposed on the respiratory muscles. Long-term home NIMV was provided to a total of 12 children with laryngomalacia (including these five) and was associated with clinical improvement in sleep and growth. PMID- 11734440 TI - Exhaled nitric oxide decreases during exercise-induced bronchoconstriction in children with asthma. AB - Nitric oxide (NO) produced in the airways can be either detrimental or protective to the host. To investigate the role of NO in the pathogenesis of exercise induced bronchoconstriction (EIB), we measured exhaled NO (ENO) after exercise challenge in 39 asthmatic and six normal children. FEV(1) and ENO were measured before and at 0, 5, 10, and 15 min after exercise performed on a treadmill for 6 min. EIB was defined as a decrease in FEV(1) of more than 15% after the exercise. Normal children (control group) did not have EIB. Twenty-one patients with asthma had EIB (EIB group) whereas the remaining 18 patients did not (non-EIB group). The baseline ENO value was significantly higher in the asthmatic children than in the normal children, and there was a positive correlation between the maximal percent decrease in FEV(1) and the baseline ENO value (r = 0.501, p = 0.012). At the end of the exercise, ENO had decreased in all the subjects. In the non-EIB and control groups, ENO rebounded to above the baseline at 5 min after the exercise and thereafter. In contrast, ENO remained at a decreased level in the EIB group. The change in ENO did not correlate with the change in minute ventilation, and beta-agonist inhalation at the peak of EIB that accelerated the recovery of FEV(1) did not affect the depressed level of ENO, demonstrating that the reduction of ENO is not a simple consequence of increased ventilation nor airway obstruction. Among the EIB group, steroid-treated patients showed sooner recovery in ENO after the exercise than steroid-naive patients. Our study suggests that NO production in response to exercise may be impaired in patients with EIB, and that ENO represents not only airway inflammation but also a protective function of NO in EIB. PMID- 11734442 TI - Renal effects of nitric oxide in endotoxemia. AB - Nitric oxide (NO) is postulated to play a key role in the pathophysiology of renal failure in sepsis. Whether the renal effects of increased NO are beneficial or harmful remains unclear. In a porcine model of lipopolysaccharide (LPS) induced shock, we evaluated the effect of LPS on glomerular filtration rate (GFR) and renal blood flow (RBF). We then administered the nonselective nitric oxide synthase (NOS) inhibitor N(G)-L-arginine methyl ester (L-NAME), and compared its effects on GFR and RBF with those of S-methylisothiourea (SMT), a selective NOS inhibitor, and those of saline. We postulated that SMT, by maintaining constitutive NO, would be more beneficial than either L-NAME or saline. LPS infusion decreased mean arterial pressure (MAP), and increased cardiac output, RBF, and medullary NO content. The increased RBF was diverted to the medulla. There was no evidence of renal dysfunction in the saline-resuscitated group. Both NOS inhibitors increased MAP but decreased RBF, but only L-NAME reduced GFR and increased sodium excretion and renal oxygen extraction. We conclude that NO in endotoxemia is beneficial because it maintains RBF and GFR. Additionally, selective NOS inhibition did not offer any advantages over saline resuscitation. PMID- 11734441 TI - Clinical characteristics of patients in a case control study of sarcoidosis. AB - Sarcoidosis may be affected by sex, race, and age. A Case Control Etiologic Study of Sarcoidosis (ACCESS) enrolled 736 patients with sarcoidosis within 6 mo of diagnosis from 10 clinical centers in the United States. Using the ACCESS sarcoidosis assessment system, we determined organ involvement for the whole group and for subgroups differentiated by sex, race, and age (less than 40 yr or 40 yr and older). The study population was heterogeneous in terms of race (53% white, 44% black), sex (64% female, 36% male), and age (46% < 40 yr old, 54% > or = 40 yr old). Women were more likely to have eye and neurologic involvement (chi(2) = 4.74, p < 0.05 and chi(2) = 4.60, p < 0.05 respectively), have erythema nodosum (chi(2) = 7.28, p < 0.01), and to be age 40 yr or over (chi(2) = 6.07, p < 0.02) whereas men were more likely to be hypercalcemic (chi(2) = 7.38, p < 0.01). Black subjects were more likely to have skin involvement other than erythema nodosum (chi(2) = 5.47, p < 0.05), and eye (chi(2) = 13.8, p < 0.0001), liver (chi(2) = 23.3, p < 0.0001), bone marrow (chi(2) = 18.8, p < 0.001), and extrathoracic lymph node involvement (chi(2) = 7.21, p < 0.01). We conclude that the initial presentation of sarcoidosis is related to sex, race, and age. PMID- 11734443 TI - Cytokine balance in the lungs of patients with acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) involves an intense inflammatory response in the lungs, with accumulation of both pro- and antiinflammatory cytokines in bronchoalveolar lavage fluid (BALF). Our goal was to determine how the balance between pro- and antiinflammatory mediators in the lungs changes before and after the onset of ARDS. We identified 23 patients at risk for ARDS and 46 with established ARDS and performed serial bronchoalveolar lavage (BAL). We used immunoassays to measure tumor necrosis factor alpha (TNF-alpha) and soluble TNF-alpha receptors I and II; interleukin 1 beta (IL-1 beta), IL-1 beta receptor antagonist, and soluble IL-1 receptor II; IL-6 and soluble IL-6 receptor; and IL-10. We used sensitive bioassays to measure net TNF-alpha, IL-1 beta, and IL-6 activity. Although individual cytokines increased before and after onset of ARDS, greater increases occurred in cognate receptors and/or antagonists, so that molar ratios of agonists/antagonists declined dramatically at the onset of ARDS. The molar ratios remained low for 7 d or longer, limiting the activity of soluble IL-1 beta and TNF-alpha in the lungs at the onset of ARDS. This significant antiinflammatory response early in ARDS may provide a key mechanism for limiting the net inflammatory response in the lungs. PMID- 11734444 TI - An algorithm to stratify sleep apnea risk in a sleep disorders clinic population. AB - Obstructive sleep apnea may lead to complications if not identified and treated. Polysomnography is the diagnostic standard, but is often inaccessible due to bed shortages. A system that facilitates prioritization of patients requiring sleep studies would thus be useful. We retrospectively compared the accuracy of a two stage risk-stratification algorithm for sleep apnea using questionnaire plus nocturnal pulse oximetry against using polysomnography to identify patients without apnea (Objective 1) and those with severe apnea (Objective 2). Patients were those referred to a university-based sleep disorders clinic due to suspicion of sleep apnea. Subjects completed a sleep apnea symptom questionnaire, and underwent oximetry and two-night polysomnography. We used bootstrap methodology to maximize sensitivity of our model for Objective 1 and specificity for Objective 2. We calculated sensitivity, specificity, positive and negative predictive values, and rate of misclassification error of the two-stage risk stratification algorithm for each of our two objectives. The model identified cases of sleep apnea with 95% sensitivity and severe apnea with 97% specificity. It excluded only 8% of patients from sleep studies, but prioritized up to 23% of subjects to receive in-laboratory studies. Among sleep disorders clinic referrals, a two-stage risk-stratification algorithm using questionnaire and nocturnal pulse oximetry excluded few patients from sleep studies, but identified a larger proportion of patients who should receive early testing because of their likelihood of having severe disease. PMID- 11734445 TI - Sleep-disordered breathing and coronary artery disease: long-term prognosis. AB - The evidence linking sleep-disordered breathing to increased mortality and cardiovascular morbidity has been conflicting and inconclusive. We hypothesized that a potential adverse effect of disordered breathing would be more obvious in patients with established vascular disease. In a prospective cohort study 408 patients aged 70 yr or younger with verified coronary disease were followed for a median period of 5.1 yr. An apnea-hypopnea index (AHI) of > or = 10 and an oxygen desaturation index (ODI) of > or = 5 were used as the diagnostic criteria for sleep-disordered breathing. The primary end point was a composite of death, cerebrovascular events, and myocardial infarction. There was a 70% relative increase and a 10.7% absolute increase in the primary composite end point in patients with disordered breathing defined as an ODI of > or = 5 (risk ratio 1.70, 95% confidence interval [CI] 1.15-2.52, p = 0.008). Similarly, patients with an AHI of > or = 10 had a 62% relative increase and a 10.1% absolute increase in the composite endpoint (risk ratio 1.62, 95% CI 1.09-2.41, p = 0.017). An ODI of > or = 5 and an AHI of > or = 10 were both independently associated with cerebrovascular events (hazard ratio 2.62, 95% CI 1.26-5.46, p = 0.01, and hazard ratio 2.98, 95% CI 1.43-6.20, p = 0.004, respectively). We conclude that sleep-disordered breathing in patients with coronary artery disease is associated with a worse long-term prognosis and has an independent association with cerebrovascular events. PMID- 11734446 TI - Accuracy of nasal cannula pressure recordings for assessment of ventilation during sleep. AB - Nasal prong pressure monitoring (PNOSE) is utilized to assess ventilation during sleep. However, it has not been rigorously validated against the gold standard of face-mask pneumotachography (VFM). Therefore, we compared PNOSE with VFM in 20 patients with suspected sleep apnea during nocturnal polysomnography, and analyzed factors affecting accuracy of PNOSE-derived variables. Patients rated their nasal obstruction on a visual analog scale. Mean +/- SE apnea/hypopnea index (AHI) by VFM was 24.0 +/- 5.1 h(-1). The bias (mean difference) and limits of agreement (+/- 2 SD) of AHI derived from PNOSE, and square root-transformed PNOSE, a measure proposed as a surrogate of airflow, were +3.9 (+/- 4.6), and 0.9 (+/- 9.0) h(-1). Subjective scores of nasal obstruction before polysomnographies did not herald inaccuracy of AHI from PNOSE. Square root transformed PNOSE closely tracked pneumotachographic airflow over 10 breaths (r(2) among signals 0.88 to 0.96) but the relationship among these signals was highly variable if comparisons were extended over an entire night. Compared with face-mask pneumotachography, nasal pressure monitoring provides accurate AHI for clinical purposes even in patients perceiving nasal obstruction. Square-root transformation provides near linear nasal pressure/airflow relationships over a short time but is not essential for estimation of AHI. PMID- 11734447 TI - Roles of mechanical forces and collagen failure in the development of elastase induced emphysema. AB - Emphysema causes a permanent destruction of alveolar walls leading to airspace enlargement, loss of elastic recoil, decrease in surface area for gas exchange, lung hyperexpansion, and increased work of breathing. The most accepted hypothesis of how emphysema develops is based on an imbalance of protease and antiprotease activity leading to the degradation of elastin within the fiber network of the extracellular matrix. Here we report novel roles for mechanical forces and collagen during the remodeling of lung tissue in a rat model of elastase-induced emphysema. We have developed a technique to measure the stress strain properties of tissue sections while simultaneously visualizing the deformation of the immunofluorescently labeled elastin-collagen network. We found that in the elastase treated tissue significant remodeling leads to thickened elastin and collagen fibers and during stretching, the newly deposited elastin and collagen fibers undergo substantially larger distortions than in normal tissue. We also found that the threshold for mechanical failure of collagen, which provides mechanical stability to the normal lung, is reduced. Our results indicate that mechanical forces during breathing are capable of causing failure of the remodeled extracellular matrix at loci of stress concentrations and so contribute to the progression of emphysema. PMID- 11734448 TI - The integrative membrane properties of human bronchial parasympathetic Ganglia neurons. AB - Parasympathetic ganglia neurons in the lower airway of laboratory animals have membrane properties associated with integration of signals from the central nervous system. In this study, intracellular recordings were made from parasympathetic ganglia located on bronchi from human lungs in order to determine the level of integration provided by human neurons. Ganglion neurons were characterized as either tonic or phasic: tonic neurons responded with repetitive action potentials sustained throughout a depolarizing current step whereas phasic neurons generated one action potential and accommodated. Phasic neurons could be further differentiated as having either short or long duration after hyperpolarizing potentials following single action potentials. In phasic neurons, stimulation of preganglionic nerves elicited one or two populations of nicotinic fast excitatory postsynaptic potentials (fEPSPs) that were graded in amplitude, subthreshold for action potential generation, and decreased in amplitude during higher frequency stimulation. In tonic neurons, single preganglionic stimuli evoked two to five populations of fEPSPs, one to three of which were at threshold for action potential generation. Dye injection into the neurons revealed multiple, branching dendrites. These results provide evidence that human bronchial ganglion neurons have unique membrane properties and anatomical characteristics associated with integrating presynaptic stimuli. Changes in these properties may thus affect output from these ganglia and, consequently, autonomic tone in the lower airways. PMID- 11734449 TI - Induction of heme-oxygenase-1 prevents the systemic responses to hemorrhagic shock. AB - Oxidant-mediated reperfusion injury of the gut is a major contributor of the systemic inflammatory response in hemorrhagic shock. Recent studies have suggested that heme-oxygenase-1 (HO-1) represents an endogenous protective mechanism against oxidant stress. We assessed whether HO-1 induction modulates the synthesis of tumor necrosis factor-alpha (TNF-alpha) in hemorrhagic shock. In rats submitted to hemorrhagic shock, pretreatment with hemoglobin (Hb) increased HO-1 mRNA expression in macrophages. This increased expression was associated with a decreased expression of TNF-alpha mRNA, as well as decreased plasma concentrations of TNF-alpha. These effects of Hb were reduced by the HO-1 inhibitor tin-protoporphyrin (Sn-PP 20 micromol/kg), while Sn-PP had no effect in the absence of Hb. In parallel, Hb pretreatment reduced pulmonary edema, vascular injury, and increased mesenteric blood flow, and these effects were reduced by Sn PP. Thus, induction of HO-1 is protective in hemorrhagic shock, possibly through its antioxidant properties. Interventions that induce HO-1 may be beneficial in the treatment of shock states, leading to a reduced systemic inflammatory response. PMID- 11734450 TI - Apoptosis of airway epithelial cells induced by corticosteroids. AB - Damage to the airway epithelium is one prominent feature of chronic asthma. Corticosteroids induce apoptosis in inflammatory cells, which in part explains their ability to suppress airway inflammation. However, corticosteroid therapy does not necessarily reverse epithelial damage. We hypothesized that corticosteroids may induce airway epithelial cell apoptosis as one potential explanation for persistent damage. We tested this hypothesis in cultured primary central airway epithelial cells and in the cell line 1HAEo(-). Treatment with dexamethasone, beclomethasone, budesonide, or triamcinolone each elicited a time dependent and concentration-dependent cell death. This cell death was associated with cleavage of nuclear chromatin, mitochondrial depolarization, cytochrome c extrusion, activation of caspase-9, and expression of phosphatidylserine on the outer cell membrane. Inhibitors of caspase activity blocked apoptotic cell death, as did overexpression of the apoptosis regulators Bcl-2 or Bcl-x(L). We demonstrated that CD95 ligation is not essential for the corticosteroid-induced apoptosis in airway epithelial cells. These data demonstrate that corticosteroids induce apoptotic cell death of airway epithelium. This raises the possibility that at least one of the major components of chronic airway damage in asthma, epithelial shedding and denudation, may in part result from a major therapy for the disease. PMID- 11734451 TI - Lung tissue eosinophils may be cleared through luminal entry rather than apoptosis: effects of steroid treatment. AB - Spontaneous or steroid-induced eosinophil apoptosis occurring in vitro has not been demonstrated in lung tissues in vivo. This study examines cell apoptosis in rat lungs using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) technique and transmission electron microscopy (TEM). After establishing sustained lung edema and eosinophilia by challenge with Sephadex beads intratracheally, budesonide treatment was started intratracheally. Sephadex alone increased the total number of apoptotic cells, which were not efficiently engulfed by macrophages or other cells, in vivo. Yet apoptotic tissue eosinophils were exceedingly rare (1 of 360 TEM-analyzed eosinophils). By contrast, approximately 20% of eosinophils in the airway lumen were apoptotic, and unengulfed. Budesonide promptly inhibited edema but 3 d of steroid treatment were required to reduce the established tissue eosinophilia. Not at any time point did budesonide induce eosinophil apoptosis (0 of 318 TEM analyzed tissue eosinophils). We conclude that (1) eosinophil apoptosis can occur but is a rare event in vivo in respiratory tract tissues; (2) airway tissue eosinophils, rather than undergoing apoptosis, are eliminated by migration into airway lumen followed by apoptosis and mucociliary clearance; (3) anti inflammatory steroid treatment may not increase eosinophil apoptosis in vivo nor may it affect the luminal entry of eosinophils; (4) steroids permit elimination of eosinophils into airway lumen and slowly resolve established lung eosinophilia. PMID- 11734452 TI - Exposure to diesel exhaust exacerbates allergen-induced airway responses in guinea pigs. AB - Diesel exhaust (DE) is a major air pollutant in urban areas. To clarify the effects of DE on the exacerbation of asthma, guinea pigs were exposed 12 h daily to 3 mg/m(3) DE or air for 8 wk with or without sensitization to ovalbumin (OVA). In the DE-exposed sensitized animals, both immediate (IAR) and late (LAR) airway responses were enhanced after the inhalation challenge by OVA, compared with the DE-unexposed sensitized animals. Mucus was greatly accumulated in the airways of DE-exposed sensitized animals during IAR. The number of eosinophils and level of sialic acid concentration in bronchial lavage fluids were also significantly higher in the DE-exposed sensitized animals than in the DE-unexposed control animals. During LAR, intercellular spaces of the bronchial epithelium became enlarged in the DE-exposed sensitized animals, showing infiltration by numerous eosinophils. Albumin concentration was significantly higher in the bronchial lavage fluids from the DE-exposed sensitized animals than in those from the DE unexposed control animals. These results suggest that exposure to DE enhances mucus hypersecretion and eosinophilic inflammation during IAR. DE exposure also increases airway permeability and airway inflammation during LAR. Thus, DE exposure exacerbates allergen-induced airway responses in guinea pigs. PMID- 11734453 TI - Induced sputum derives from the central airways: confirmation using a radiolabeled aerosol bolus delivery technique. AB - Indirect evidence suggests that induced sputum derives from the surfaces of the bronchial airways. To confirm this experimentally, we employed a radiolabeled aerosol bolus delivery technique that preferentially deposits aerosol in the central airways in humans. We hypothesized that there would be significantly more radioactivity recovered in an induced sputum sample, and greater airways clearance of radiolabeled particles, immediately after a central versus peripheral airways deposition. Ten healthy volunteers underwent radiolabeled aerosol deposition ((99m)Tc sulfur colloid particles) to the central and peripheral airways on separate occasions followed immediately by induced sputum or no sputum (control), while seated in front of a gamma camera. Radioactivity was measured in the selected sputum sample, processed cell pellet, and supernatant fraction. Significantly more radioactivity was present in all portions of the sputum sample after central versus peripheral airways deposition (i.e., selected sample: 15,607 counts +/- 2,985 versus 943 counts +/- 298, p = 0.001). Clearance from the whole lung was significantly greater 40 min after central versus peripheral airways deposition (48 +/- 3% versus 5 +/- 1%, p = 0.0001). Compared with control, induced sputum greatly enhanced clearance after central deposition (48 +/- 3% versus 11 +/- 6%, p = 0.0001), but not after peripheral deposition (5 +/- 1% versus 3 +/- 0.8%). These results provide direct evidence that induced sputum derives from the central airways with little or no contribution from the peripheral airways. PMID- 11734454 TI - Disrupted pulmonary vasculature and decreased vascular endothelial growth factor, Flt-1, and TIE-2 in human infants dying with bronchopulmonary dysplasia. AB - An abnormal pulmonary vasculature may be an important component of bronchopulmonary dysplasia (BPD). We examined human infant lung for the endothelial cell marker PECAM-1 and for angiogenic factors and their receptors. Lung specimens were collected prospectively at approximately 6 h after death. The right middle lobe was inflation fixed and part of the right lower lobe was flash frozen. We compared lungs from infants dying with BPD (n = 5) with lungs from infants dying from nonpulmonary causes (n = 5). The BPD group was significantly more premature and had more days of ventilator and supplemental oxygen support, but died at a postconceptional age similar to control infants. PECAM-1 protein and mRNA were decreased in the BPD group. PECAM-1 immunohistochemistry showed the BPD group had decreased staining intensity and abnormal distribution of alveolar capillaries. The dysmorphic capillaries were frequently in the interior of thickened alveolar septa. The BPD group had decreased vascular endothelial growth factor (VEGF) mRNA and decreased VEGF immunostaining, compared with infants without BPD. Messages for the angiogenic receptors Flt-1 and TIE-2 were decreased in the BPD group. We conclude that infants dying with BPD have abnormal alveolar microvessels and that disordered expression of angiogenic growth factors and their receptors may contribute to these abnormalities. PMID- 11734455 TI - Pulmonary vascular endothelial growth factor and Flt-1 in fetuses, in acute and chronic lung disease, and in persistent pulmonary hypertension of the newborn. AB - Respiratory distress syndrome (RDS) and development of bronchopulmonary dysplasia (BPD) are characterized by endothelial cell damage. Persistent pulmonary hypertension of the newborn (PPHN) is a disorder that alters the pulmonary microvasculature. Immunohistochemistry for VEGFA(165), an endothelial cell mitogen, and its receptor Flt-1, was performed on lung tissues from autopsies from four fetuses, three preterm infants, four term infants without primary lung disease, four infants with BPD, and four infants with PPHN. VEGF was measured in tracheal aspirates from 31 preterm infants, 5 intubated term infants without primary lung injury, and 12 infants with PPHN during the first 10 postnatal days, and from 8 infants with BPD. Immunohistochemistry for VEGF and Flt-1 was similar in fetuses, preterm infants, and term infants: for VEGF mostly in bronchial epithelium and alveolar macrophages, and for Flt-1 mostly in vascular endothelial cells and bronchial epithelial cells. In patients with BPD, and PPHN, staining for VEGF and Flt-1 appeared also in Type II pneumocytes. Preterm infants with more severe RDS had lower VEGF than those who recovered. The persistent expression of VEGF and Flt-1 during the fetal and neonatal period supports a physiological role for VEGF in human lung development. The lower pulmonary VEGF in preterm infants with more severe RDS may contribute to the pathophysiology of the acute lung injury. In BPD, the expression of VEGF in alveolar epithelium may represent a compensatory increase after the acute phase of the lung disease. In PPHN, that more cell types express VEGF and Flt-1, and the tendency toward a higher concentration of pulmonary VEGF may represent enhanced production of VEGF in response to impaired endothelial function. PMID- 11734456 TI - Coagulation blockade prevents sepsis-induced respiratory and renal failure in baboons. AB - Sepsis-induced tissue factor (TF) expression activates coagulation in the lung and leads to a procoagulant environment, which results in fibrin deposition and potentiates inflammation. We hypothesized that preventing initiation of coagulation at TF-Factor VIIa (FVIIa) complex would block fibrin deposition and control inflammation in sepsis, thereby limiting acute lung injury (ALI) and other organ damage in baboons. A model of ALI was used in which adult baboons were primed with killed Escherichia coli (1 x 10(9) CFU/kg), and bacteremic sepsis was induced 12 h later by infusion of live E. coli at 1 x 10(10) CFU/kg. Animals in the treatment group were given a competitive inhibitor of TF, site inactivated FVIIa (FVIIai), intravenously at the time of the infusion of live bacteria and monitored physiologically for another 36 h. FVIIai dramatically protected gas exchange and lung compliance, prevented lung edema and pulmonary hypertension, and preserved renal function relative to vehicle (all p < 0.05). Treatment attenuated sepsis-induced fibrinogen depletion (p < 0.01) and decreased systemic proinflammatory cytokine responses, for example, interleukin 6 (p < 0.01). The protective effects of TF blockade in sepsis-induced ALI were confirmed by using tissue factor pathway inhibitor. The results show that TF-FVIIa complex contributes to organ injury in septic primates in part through selective stimulation of proinflammatory cytokine release and fibrin deposition. PMID- 11734458 TI - Serum nitrite/nitrate levels in obstructive sleep apnea. PMID- 11734459 TI - Therapeutic ratio of inhaled corticosteroids: fact or fiction? PMID- 11734460 TI - Efficiency and safety of mechanical ventilation with a heat and moisture exchanger changed once a week. PMID- 11734463 TI - Chairman's summary. Mechanisms of airway remodeling. PMID- 11734464 TI - Remodeling in asthma and chronic obstructive lung disease. AB - Asthma and chronic obstructive lung disease (COPD) are both inflammatory conditions of the lung associated with structural "remodeling" inappropriate to the maintenance of normal lung function. The clinically observed distinctions between asthma and COPD are reflected by differences in the remodeling process, the patterns of inflammatory cells and cytokines, and also the predominant anatomic site at which these alterations occur. In asthma the epithelium appears to be more fragile than that of COPD, the epithelial reticular basement membrane (RBM) is significantly thicker, there is marked enlargement of the mass of bronchial smooth muscle, and emphysema does not occur in the asthmatic nonsmoker. In COPD, there is epithelial mucous metaplasia, airway wall fibrosis, and inflammation associated with loss of surrounding alveolar attachments to the outer wall of small airways: bronchiolar smooth muscle is increased also. Emphysema is a feature of severe COPD: in spite of the destructive process, alveolar wall thickening and focal fibrosis may be detected. The hypertrophy of submucosal mucus-secreting glands is similar in extent in asthma and COPD. The number of bronchial vessels and the area of the wall occupied by them increase in severe corticosteroid-dependent asthma: it is likely that these increases also occur in severe COPD as they do in bronchiectasis. Pulmonary vasculature is remodeled in COPD. In asthma several of these structural alterations begin early in the disease process, even in the child. In COPD the changes begin later in life and the associated inflammatory response differs from that in asthma. The following synopsis defines and compares the key remodeling processes and proposes several hypotheses. PMID- 11734465 TI - Angiogenesis and remodeling of airway vasculature in chronic inflammation. AB - Angiogenesis and microvascular remodeling are known features of chronic inflammatory diseases such as asthma and chronic bronchitis, but the mechanisms and consequences of the changes are just beginning to be elucidated. In a model of chronic airway inflammation produced by Mycoplasma pulmonis infection of the airways of mice or rats, angiogenesis and microvascular remodeling create vessels that mediate leukocyte influx and leak plasma proteins into the airway mucosa. These vascular changes are driven by the immune response to the organisms. Plasma leakage results from gaps between endothelial cells, as well as from increased vascular surface area and probably other changes in the newly formed and remodeled blood vessels. Treatment with long-acting beta2 agonists can reduce but not eliminate the plasma occurring after infection. In addition to the elevated baseline leakage, the remodeled vessels in the airway mucosa are abnormally sensitive to substance P, but not to platelet-activating factor or serotonin, suggesting that the infection leads to a selective upregulation of NK1 receptors on the vasculature. The formation of new vessels and the remodeling of existing vessels are likely to be induced by multiple growth factors, including vascular endothelial growth factor (VEGF) and angiopoietin 1 (Ang1). VEGF increases vascular permeability, but Ang1 has the opposite effect. This feature is consistent with evidence that VEGF and Ang1 play complementary and coordinated roles in vascular growth and remodeling and have powerful effects on vascular function. Regulation of vascular permeability by VEGF and Ang1 may be their most rapid and potent actions in the adult, as these effects can occur independent of their effects on angiogenesis and vascular remodeling. The ability of Ang1 to block plasma leakage without producing angiogenesis may be therapeutically advantageous. Furthermore, because VEGF and Ang1 have additive effects in promoting angiogenesis but opposite effects on vascular permeability, they could be used together to avoid the formation of leaky vessels in therapeutic angiogenesis. Finally, the elucidation of the protective effect of Ang1 on blood vessel leakiness to plasma proteins raises the possibility of a new strategy for reducing airway edema in inflammatory airway diseases such as asthma and chronic bronchitis. PMID- 11734466 TI - Remodeling of the airway epithelium in asthma. AB - Several pathologic changes occur in the airway epithelium in asthma, but the relationship between these changes and the initiation and progression of asthma remains poorly understood. One possibility is that changes in the structure and function of the epithelium induced by environmental exposure in genetically susceptible subjects represent primary pivotal events that occur early in the pathogenesis of asthma. Alternatively, these epithelial changes may occur simply as a consequence of pivotal early events in other systems, such as immune deviation in childhood to a helper T cell type 2 (Th2) subtype of CD4(+) cells. Epithelial desquamation in asthma represents a pathologic change that is frequently cited as important for the mechanisms of airway remodeling and airway hyperresponsiveness. Desquamation of the epithelium may not represent true pathology, however, but may instead be an artifact of tissue sampling and handling. Evidence is more firm for other pathologic changes in the epithelium. For example, goblet cell numbers are increased in asthma, leading to increases in stored mucins in the epithelium and in secreted mucins in sputum. The functional consequences of these changes include sputum production and airway narrowing, which lead to asthma exacerbations. Currently available data suggest that an important mechanism for goblet cell hyperplasia in asthma is the action of Th2 cytokines. Improved understanding of epithelial goblet cell abnormalities in asthma will hopefully lead to novel therapies for mucin hypersecretion, which is an important cause of morbidity and mortality. PMID- 11734467 TI - Mast cell tryptases and airway remodeling. AB - On the basis of their amino acid sequences, tryptases are just another group of serine proteinases related to trypsin that happen to be expressed and stored in mast cells rather than the pancreas. On the basis of their biochemical and biological features, however, tryptases show little family likeness to trypsin and most other trypsin-like proteases. The intriguing discrepancies have been explained by the crystal structure of the tryptase tetramer. It is now clear how tryptases, by forming tetramers, have gained the ability to prevail enzymatically active in tissues, but, at the cost of an unusual narrow substrate specificity. The tryptase tetramer thus became both a (neuro)peptidase and a long-lasting initiator that orchestrates responses by the cleavage of a few key proteins, the activation of other proteases with broader specificity, and the stimulation of cellular responses. With the support of these performers, tryptase drives a variety of processes contributing to chronic inflammation and tissue remodeling, the diversity of which is still emerging. PMID- 11734468 TI - Do lung remodeling, repair, and regeneration recapitulate respiratory ontogeny? AB - Herein we posit that modeling of the lungs during morphogenesis, repair, and regeneration is tightly coordinated by conserved stimulatory and inhibitory signaling mechanisms, including specific transcriptional factors, cytokines, peptide growth factors, proteases, and matrix elements. This evolutionary developmental (evo-devo) functional conservation has been extended to morphogenesis of the respiratory tracheae in Drosophila. Fifty or more genes direct fruit fly tracheal organogenesis. Among them, hedgehog, patched, smoothened, cubitus interruptus, branchless, breathless, sprouty, decapentaplegic, and mad are functionally conserved between flies, mice, and humans. For example, fibroblast growth factor (FGF) signaling is essential, not only for fly trachea and mouse bronchial branching morphogenesis, but also for postnatal modeling and repair of alveoli. Likewise, sprouty family genes act as inducible negative regulators of FGF signaling, which in part may determine interbranch length during bronchial development. Alveolar epithelial survival, migration, and proliferation during remodeling after hyperoxic injury also require FGF signaling. In addition, FGF signaling appears to regulate a small (< 5%) population of putative alveolar stem/ progenitor cells that express telomerase and are relatively resistant to hyperoxic apoptosis. We speculate that genes in evo-devo functionally conserved signaling pathways such as FGF-FGF receptor-Sprouty may provide novel therapeutic targets to augment lung repair and induce lung regeneration. PMID- 11734469 TI - Mechanisms of airway remodeling. Airway smooth muscle. AB - The airway smooth muscle cell can contract; relax; participate in allergic and inflammatory responses by expressing adhesion molecules, releasing cytokines, and producing matrix proteins and proteases; and, as has been reported, undergo migration. These properties enable the muscle cell to be a key component in the airway wall remodeling that accompanies persistent asthma. Evidence is emerging that identifies the pivotal steps in the signal transduction pathways that lead to the excessive proliferation of the muscle observed in vitro in airway smooth muscle cells from subjects with asthma. The contractile, biochemical, and growth characteristics of muscle from allergic subjects are different from those of nonallergic subjects. In addition, the allergic response impacts on the extracellular matrix in which the muscle is embedded, by altering the profile of matrix proteins released. Once the relationships between allergy and inflammation of the smooth muscle and its extracellular matrix are better defined, opportunities to prevent or reverse airway remodeling will become available. PMID- 11734470 TI - Airway inflammation and remodeling in asthma. Lessons from interleukin 11 and interleukin 13 transgenic mice. AB - Noninflammatory structural alterations, variously referred to as airway remodeling, are well documented in the asthmatic airway. However, the pathogenesis of these alterations, the importance of airway remodeling in generating the asthma phenotype, and the natural history of airway remodeling responses have not been adequately defined. Because exaggerated cytokine production is a characteristic feature of the asthmatic airway, we used constitutive and inducible overexpression transgenic systems to investigate the contributions that interleukin 11 (IL-11) and IL-13 might make to airway remodeling responses. These studies demonstrated that both cytokines produce responses in the murine airway with features similar to those in human asthmatic tissues. IL-11 caused airway fibrosis with the enhanced accumulation of interstitial collagens, myocytes, and myofibroblasts. IL-13 caused mucous metaplasia, enhanced mucin gene expression, enhanced tissue hyaluronic acid accumulation, and subepithelial fibrosis. Importantly, IL-11 was detected most readily in tissues from asthmatic subjects with severe airway remodeling that was similar to that seen in the IL-11 transgenic mice. In addition, IL-11 was shown to inhibit asthma-like inflammation while stimulating airway fibrosis. This suggests that IL-11 elaboration is, in part, an attempt at airway healing. Last, a novel triple transgenic system is described that allows transgene expression to be regulated in a true "on/off" manner. This system may be useful in defining the reversibility of transgene-induced airway remodeling responses. PMID- 11734471 TI - Role of latent viral infections in chronic obstructive pulmonary disease and asthma. AB - Acute viral respiratory tract infections are well known to precipitate asthma attacks and acute exacerbations of chronic obstructive pulmonary disease, but their role in the pathogenesis of chronic disease is poorly defined. Double stranded DNA viruses have the ability to persist in airway epithelial cells long after the acute infection has cleared. During these latent infections, viral genes are expressed at the protein level without replication of a complete virus. The expression of the adenoviral trans-activating protein has been demonstrated in the airway epithelium of both human and animal lungs and is associated with an amplification of the cigarette smoke-induced inflammatory response. Studies of cultured human airway epithelial cells have also shown that transfection with this viral gene upregulates the expression of intercellular adhesion molecule 1 and interleukin 8 by these cells when they are challenged with endotoxin. In guinea pigs, cigarette smoke-induced emphysema is amplified by latent adenoviral infection. Furthermore, this infection independently increased the number of CD-8 cells, whereas the cigarette smoke independently increased the number of CD-4 cells in the inflammatory infiltrate. On the other hand, allergen-induced lung inflammation was uninfluenced by latent adenoviral infection in the guinea pig, but the latent infection caused the eosinophilic component of this response to become steroid resistant. These studies suggest that latent adenoviral infections may have a role in the pathogenesis of obstructive airway disease by amplifying the response to cigarette smoke and inducing steroid resistance. PMID- 11734472 TI - Remodeling in response to infection and injury. Airway inflammation and hypersecretion of mucus in smoking subjects with chronic obstructive pulmonary disease. AB - Airway epithelium represents the first line of defense against toxic inhalants. In some subjects, cigarette smoking causes airway inflammation, hypersecretion of mucus, and poorly reversible airflow limitation through mechanisms that are still largely unknown. Likewise, it is unclear why only some smokers develop chronic obstructive pulmonary disease (COPD). Two cell types consistently result in relation to chronic airflow limitation in COPD: neutrophils and CD8(+) cells. Neutrophils are compartmentalized in the mucosal surface of the airways and air spaces, that is, the epithelium and lumen, whereas CD8(+) cells exhibit a more extensive distribution along the subepithelial zone of the airways and lung parenchyma, including alveolar walls and arteries. This pattern of inflammatory cell distribution is observed in mild or moderate COPD, and in patients who have developed COPD, it is not modified by smoking cessation. The number of neutrophils further increases in the submucosa of patients with severe COPD, suggesting a role for these cells in the progression of the disease. Hypersecretion of mucus is a major manifestation in COPD. Mucus is produced by bronchial glands and goblet cells lining the airway epithelium. Unlike mucous gland enlargement, greater mucosal inflammation is associated with sputum production. Whereas neutrophil infiltration of submucosal glands occurs only in smokers with COPD, goblet cell hyperplasia in peripheral airways occurs both in smokers with or without COPD, suggesting that the major determinant of goblet cell hyperplasia is cigarette smoke itself. PMID- 11734473 TI - Role of cyclins in epithelial response to oxidants. AB - Oxidants are involved in a large variety of pulmonary diseases. Among the various cell types that compose the respiratory system, the epithelial cells appear to be a major target for oxidative stress. When cells are exposed to DNA-damaging agents such as oxidants, a feedback control is activated that acts as a brake on the cell cycle to inhibit entry into the S phase until DNA repair is completed. Progression through the G1 phase and the G1-S transition involves sequential assembly and activation of key regulators of the cell cycle machinery, the cyclin dependent kinases (CDKs). Activity of the CDKs is regulated by several mechanisms, which include the CDK inhibitors (CKIs). The CKI p21(CIP1) appears to play an important role in the response of epithelial cells to oxidants. PMID- 11734474 TI - Development of high-density DNA microarray membrane for profiling smoke- and hydrogen peroxide-induced genes in a human bronchial epithelial cell line. AB - Development of the high-density DNA microarray technique permits the analysis of thousands of genes simultaneously for their differential expression patterns in various biological processes. Through clustering analysis and pattern recognition, the significance of differentially expressed genes can be recognized and correlated with biological events that may take place inside the cell and tissue. With this notion in mind, high-density DNA microarray nylon membrane with colorimetry detection was used to profile the expression of smoke- and hydrogen peroxide-inducible genes in a human bronchial epithelial cell line, HBE1. On the basis of the time course of expression, at least three phases of change in gene expression could be recognized. The first phase is an immediate event in response to oxidant injury. This phase includes induction of the bcl-2 and mdm-2 genes, which are involved in the regulation of apoptosis, and the mitogen-activated protein (MAP) kinase phosphatase 1 (MKP-1) gene, that functions as a regulator of various mitogen-activated protein kinase activities. The second phase, usually 5 h later, includes the induction of various stress proteins and ubiquitin, which are important in providing the chaperone mechanism and the turnover of damaged macromolecules. The third phase, which is 5-10 h later, includes the induction of genes that are apparently involved in reducing oxidative stress by metabolizing reactive oxygen species. In this phase, enzymes associated with tissue and cell remodeling are also elevated. These results demonstrate a complex gene expression array by bronchial epithelial cells in response to the insult of oxidants that are relevant to environmental pollutants. PMID- 11734475 TI - Mechanical stimuli to airway remodeling. AB - The airway is exposed to a variety of mechanical stimuli, the most prominent of which is the acute compressive stress caused by bronchoconstriction. The folding of the airway wall into a rosette pattern during bronchoconstriction creates a complex stress field, with the highest stresses compressing the epithelial layer at the inner surface of the airway wall. The epithelial cells lining the airway possess the capacity to modulate the inflammatory environment of the airway wall, and produce factors that influence the recruitment, proliferation, and activity of fibroblasts and smooth muscle cells. A variety of in vitro studies have demonstrated that airway epithelial cells, along with lung fibroblasts and smooth muscle cells, are responsive to mechanical stimuli. Airway epithelial cells exposed to compressive stresses matched to those occurring in the constricted airway increase expression of genes relevant to airway remodeling, and increase the collagen synthesis of cocultured fibroblasts. These findings demonstrate that mechanical stress may contribute to the remodeling of the asthmatic airway. PMID- 11734477 TI - Presidential address of the American Orthopaedic Society for Sports Medicine. Get involved! PMID- 11734478 TI - Augmentation of tendon healing in an intraarticular bone tunnel with use of a bone growth factor. AB - We hypothesized that an exogenous bone growth factor could augment healing of a tendon graft in a bone tunnel in a rabbit anterior cruciate ligament reconstruction model. Seventy rabbits underwent bilateral anterior cruciate ligament reconstructions with a semitendinosus tendon graft. One limb received a collagen sponge carrier vehicle containing a mixture of bone-derived proteins while the contralateral limb was treated with either no sponge or a sponge without bone-derived proteins. The reconstruction was evaluated at 2, 4, or 8 weeks with histologic, biomechanical, and magnetic resonance imaging analysis. Histologic analysis demonstrated that specimens treated with bone-derived proteins had a more consistent, dense interface tissue and closer apposition of new bone to the graft, with occasional formation of a fibrocartilaginous interface, when compared with control specimens. The treated specimens had significantly higher load-to-failure rates than did control specimens. Treatment with bone-derived proteins resulted in an average increase in tensile strength of 65%. The treated specimens were stronger than control specimens at each time point, but the difference was greatest at 8 weeks. On the basis of signal characteristics and new bone formation, magnetic resonance imaging was useful for predicting which limb was treated, the site of failure, and the limbs with higher load-to-failure values. This study demonstrates the potential for augmenting tendon healing in an intraarticular bone tunnel using an osteoinductive growth factor. PMID- 11734479 TI - A prospective evaluation of untreated acute grade III acromioclavicular separations. AB - We prospectively studied the natural history of untreated acute grade III acromioclavicular separations. Twenty-five patients were treated nonoperatively with a sling for comfort through progressive early range of motion as tolerated. Ten additional uninjured subjects underwent strength testing to evaluate the difference between the dominant and nondominant sides so that patient data could be standardized. The patients were examined at intervals of 6, 12, 24, 36, and 52 weeks after injury, at which time they completed a subjective questionnaire and underwent isometric dynamometer testing as well as military press and bench press strength testing. One patient underwent a surgical procedure at 2 weeks after injury because of cosmetic concerns. Twenty of the 25 patients completed the 1 year evaluation and strength-testing protocol. Subjectively, 4 of the 20 patients (20%) thought that their long-term outcome was suboptimal, although for 3 of them it was not enough to warrant surgery. Objective examination and strength testing of the 20 patients revealed no limitation of shoulder motion in the injured extremity and no difference between sides in rotational shoulder muscle strength. The bench press was the only strength test that showed a significant short-term difference, with the injured extremity being an average of 17% weaker. This study documents the natural history of patients with an untreated acute grade III acromioclavicular separation and provides a reference with which to judge all other proposed methods of treatment. PMID- 11734480 TI - Matrix assessment of the articular cartilage surface after chondroplasty with the holmium:yttrium-aluminum-garnet laser. A long-term study. AB - A long-term in vivo study was performed to assess biochemical changes after laser repair of articular cartilage. Forty New Zealand White rabbits were sacrificed 26 weeks after undergoing an articular cartilage chondroplasty with use of a holmium:yttrium-aluminum-garnet laser at 0.8 joules per pulse and a rate of 10 Hz. Glycosaminoglycan content in the repaired tissue decreased significantly with both perpendicular (19.59+/-5.6 microg hexosamin/mg of dry tissue) and tangential delivery (14.78+/-4.5 microg/mg) compared with the sham-treated tissue (39.6+/ 5.0 microg/mg). Cellular viability was also significantly decreased. Sulfate incorporation was decreased to 203+/-142 cpm/mg of dry cartilage in the tangential mode and 461+/-209 cpm/mg in the tangential mode, compared with the sham at 1845 cpm/mg. Uptake of [3H]thymidine decreased to 463+/-473 cpm/mg of dry tissue and 455+/-170 cpm/mg in the tangential and perpendicular modes, respectively, compared with 2465 cpm/mg in the sham tissue. There were no significant differences between the tangential and perpendicular delivery modes in any assessments performed. The shortterm chondrocyte destruction previously noted in a 12-week study after laser treatment was not reversed during a longer term 26-week study, and cellular viability was not recovered, suggesting that the loss of chondrocyte function may be permanent. PMID- 11734481 TI - Tuberculum intercondylare tibiae tertium as a predictive factor for anterior cruciate ligament injury. AB - The aim of this study was to assess the presence of tuberculum intercondylare tibiae tertium, also known as Parsons' knob, and to determine its prominence. Knee radiographs of 171 patients operated on for anterior cruciate ligament injury were examined. The control group included 120 sex- and age-matched patients who underwent orthopaedic examination for knee pain, in whom anterior cruciate ligament injury was ruled out. Knee radiographs revealed the presence of tuberculum intercondylare tibiae tertium in 55 (32.2%) and 16 (13.3%) patients from the study and control groups, respectively. The between-group difference was statistically significant. The authors developed their own method of tuberculum intercondylare tibiae tertium measurement using proportional coefficients based on the length of the tibial plateau to compare the values of the tuberculum intercondylare tibiae tertium. The results showed tuberculum intercondylare tibiae tertium to be not only more common but also more pronounced, especially in height, in the patients with anterior cruciate ligament lesions. PMID- 11734482 TI - Abnormally decreased regional bone density in athletes with medial tibial stress syndrome. AB - Medial tibial stress syndrome, a common condition of uncertain origin found in athletes, is characterized by pain in the distal posteromedial aspect of the tibia during exercise, with or without increased scintigraphic uptake in the affected region. To determine whether medial tibial stress syndrome with increased scintigraphic uptake is associated with a change in tibial bone mineral density confined to the site of the increased uptake, we measured bone mineral density (in grams per square centimeter) in 18 adult male athletes with long standing medial tibial stress syndrome and compared the measurements with those of 16 age- and sex-matched control subjects and with those of 18 athletes without medial tibial stress syndrome who had a comparable training regimen. Tibial bone mineral density in the region corresponding to the pain was 15%+/-9% lower in the patients than in control subjects and 23%+/-8% lower than in the athletic control subjects (both significant differences). Bone mineral densities in most other regions of the body were higher than in the control subjects but lower than in the athletic controls at the corresponding sites. In summary, medial tibial stress syndrome is associated with low regional bone mineral density. PMID- 11734483 TI - Relationship of ulnar collateral ligament strain to amount of medial olecranon osteotomy. AB - Athletes at risk for valgus extension overload are also at risk for tears of the anterior bundle of the ulnar collateral ligament. Some athletes develop ligament tears after procedures for valgus extension overload such as posteromedial olecranon osteotomy. The amount of posteromedial olecranon that can be resected before ulnar collateral ligament strain, and risk of injury, increases is unknown. We dissected and mounted five fresh-frozen human cadaveric elbows to allow strain gauge monitoring of the ulnar collateral ligament with varying valgus stress, elbow flexion angle, and medial osteotomy. The average strain to failure was 11.96%+/-6.51%, corresponding to a load of 347.71+/-46.42 N. The maximum tensile force recorded at failure was 416.24 N. Three-way repeated measures analysis of variance revealed no significant change in strain with change in the amount of osteotomy for a given applied load and angle of flexion. On the basis of these data, we conclude that the effect of medial olecranon osteotomy on ulnar collateral ligament strain may be small. Small sample size, elderly specimens, and the variables inherent in the experimental setup and mathematical modeling make it difficult to extrapolate these results to in vivo behavior of the anterior ulnar collateral ligament. Further work is needed before definitive guidelines for olecranon osteotomy can be formulated. PMID- 11734484 TI - Four-strand hamstring tendon autograft compared with patellar tendon-bone autograft for anterior cruciate ligament reconstruction. A randomized study with two-year follow-up. AB - Seventy-two patients with subacute or chronic rupture of the anterior cruciate ligament were randomly assigned to autograft reconstruction with four-strand gracilis and semitendinosus tendon (N = 37) or with patellar tendon-bone (N = 35) from the ipsilateral side. The groups were similar in terms of age, sex, level of activity, degree of laxity, meniscal lesions found surgically, and rehabilitation program. The follow-up was performed at another hospital by independent observers after 6, 12, and 24 months. Sixty-one patients (32 with hamstring tendon grafts and 29 with patellar tendon grafts) complied with the follow-up routine for the full 24 months. No differences were found between the groups with respect to Cincinnati functional score, KT-1000 arthrometer measurements, or stairs hopple test results. The subjective result and the single-legged hop test result were better for the hamstring tendon group after 6 and 12 months, but no differences were found after 24 months. The hamstring tendon group showed better isokinetic knee extension strength than did the patellar tendon group after 6 months, but not after 12 and 24 months. There was a significant weakness in isokinetic knee flexion strength among the hamstring tendon group. Anterior knee pain was not significantly different between the groups, but kneeling pain was significantly less common in the hamstring tendon group after 24 months. PMID- 11734485 TI - The anatomic relationship of the brachial plexus and axillary artery to the glenoid. Implications for anterior shoulder surgery. AB - Iatrogenic brachial plexus injury is an uncommon but potentially severe complication of shoulder reconstruction for instability that involves dissection near the subscapularis muscle and potentially near the brachial plexus. We examined the relationship of the brachial plexus to the glenoid and the subscapularis muscle and evaluated the proximity of retractors used in anterior shoulder surgical procedures to the brachial plexus. Eight fresh-frozen cadaveric shoulders were exposed by a deltopectoral approach. The subscapularis muscle was split in the middle and dissected to reveal the capsule beneath it. The capsule was split at midline, and a Steinmann pin was placed in the equator of the glenoid rim under direct visualization. The distance from the glenoid rim to the brachial plexus was measured with calipers with the arm in 0 degrees, 60 degrees, and 90 degrees of abduction. The brachial plexus and axillary artery were within 2 cm of the glenoid rim, with the brachial plexus as close as 5 mm in some cases. There was no statistically significant change in the distance from the glenoid rim to the musculocutaneous nerve, axillary artery, medial cord, or posterior cord with the arm in various degrees of abduction. Retractors placed superficial to the subscapularis muscle or used along the scapular neck make contact with the brachial plexus in all positions tested. PMID- 11734486 TI - Biomechanical analysis of the effect of varying suture pitch in tendon graft fixation. AB - The purpose of this study was to biomechanically assess the effect of varying suture pitch on the holding power of the Krackow suture technique for tendon graft fixation. Seven pairs of rabbit Achilles tendons were sutured with single No. 5 Ti-Cron suture using the Krackow technique. One tendon from each pair was sutured using a 0.5-cm suture pitch (half-pitch group) while the contralateral tendon from each pair was sutured with a 1.0-cm suture pitch (one-pitch group). The tendons were loaded to failure using a servohydraulic materials test system at a loading rate of 0.5 mm/sec. There were no statistically significant differences noted in suture slippage at failure (1.58 cm for half pitch versus 1.45 cm for one pitch) or maximal force to failure (158.5 N for half pitch versus 168.2 N one pitch) between the two treatment groups. However, the construct with the 1-cm suture pitch was significantly stiffer than the construct with the 0.5 cm suture pitch, with stiffness values of 106.2 N/cm and 91.4 N/cm, respectively. The most common mechanism of failure was slippage of the suture at the first suture throw and tearing of the first knot through the most distal portion of the tendon. Four constructs failed by suture rupture, two from each experimental group. PMID- 11734487 TI - The kinematics and kinetics of slipknots for arthroscopic Bankart repair. AB - An apparatus has been developed to enable the in vitro measurement of the kinematics and kinetics of a slipknot, both while the knot is being tightened and with the knot in place and the tension removed. During tightening, the apparatus provides a linearly increasing resistance, which may be considered analogous to the resistance experienced when the labrum is drawn toward and apposed to the edge of the glenoid cavity during a Bankart repair. The tension to close the knot is measured with a tensiometer, in tandem with the closure of the capsule model onto the bone anchor, which is measured with a datalogger. The tightening tension was limited to 25 N and intraoperator and interoperator comparisons were made for four knots tied by six participants. Failure of a knot was taken to be reverse slippage of 2 mm. A second criterion of ease of slide was used to assess each knot. We found wide interoperator variability with regard to slipknot tying. A methodology for quality control of slipknot tying is presented. PMID- 11734488 TI - Revision anterior cruciate ligament reconstruction with a reharvested ipsilateral patellar tendon. AB - The patellar tendon remains the most popular graft choice for anterior cruciate ligament reconstruction and has been proven to be the strongest substitute. Between 1991 and 1998, we performed revision anterior cruciate ligament reconstruction using the reharvested central third of the ipsilateral patellar tendon in 15 patients. Adequate follow-up was obtained in 13 of these 15 patients. The results in these 13 patients (mean age, 27.2 years) were reviewed. At an average postoperative follow-up of 39.4 months (range, 24 to 65), 11 patients had good or excellent results and 2 patients had fair results. Clinical examination revealed an average Tegner knee score of 5.8 (range, 3 to 9) and an average Lysholm knee score of 77.6 (range, 61 to 98). Postoperative KT-1000 arthrometer results showed an average side-to-side difference of 1.92 mm (range, 2.0 to 4.0). No patient demonstrated any loss of range of motion and only one reported patellofemoral problems, which were moderate. These favorable results demonstrate that, with appropriate patient selection, the use of a reharvested central third patellar tendon is a viable option for revision of a failed anterior cruciate ligament reconstruction. PMID- 11734490 TI - Greater tuberosity notch: an important indicator of articular-side partial rotator cuff tears in the shoulders of throwing athletes. AB - We examined the location of rotator cuff tears, associated labral injuries, and notches on the greater tuberosity of the humeral head in shoulders of throwing athletes. Arthroscopic findings (rotator cuff tear, labral condition, and greater tuberosity notch) as well as other factors (duration of playing baseball, range of motion, and joint laxity) of 61 baseball players were retrospectively studied. The presence of a greater tuberosity notch was also evaluated for by plain radiographs. Forty patients had articular-side partial rotator cuff tears, most of which occurred in the interval between the supraspinatus and infraspinatus tendons. The existence of a rotator cuff tear was not related to the range of motion, joint laxity, the detachment of the superior glenoid labrum, or posterosuperior labral injury. Greater tuberosity notches were recognized in 38 shoulders by arthroscopy and most were detected on plain radiographs. The presence of a notch was significantly related to the existence of a rotator cuff tear, while the size of the notch was significantly related to the depth and width of the tear. The greater tuberosity notch seems to be one of the most important diagnostic indicators for a rotator cuff tear in throwing athletes. PMID- 11734489 TI - Biomechanical properties and vascularity of an anterior cruciate ligament graft can be predicted by contrast-enhanced magnetic resonance imaging. A two-year study in sheep. AB - Magnetic resonance imaging has been used to determine graft integrity and study the remodeling process of anterior cruciate ligament grafts morphologically in humans. The goal of the present study was to compare graft signal intensity and morphologic characteristics on magnetic resonance imaging with biomechanical and histologic parameters in a long-term animal model. Thirty sheep underwent anterior cruciate ligament reconstruction with an autologous Achilles tendon split graft and were sacrificed after 6, 12, 24, 52, or 104 weeks. Before sacrifice, all animals underwent plain and contrast-enhanced (gadolinium diethylenetriamine pentacetic acid) magnetic resonance imaging (1.5 T, proton density weighted, 2-mm sections) of their operated knees. The signal/noise quotient was calculated and data were correlated to the maximum load to failure, tensile strength, and stiffness of the grafts. The vascularity of the grafts was determined immunohistochemically by staining for endothelial cells (factor VIII). We found that high signal intensity on magnetic resonance imaging reflects a decrease of mechanical properties of the graft during early remodeling. Correlation analyses revealed significant negative linear correlations between the signal/noise quotient and the load to failure, stiffness, and tensile strength. In general, correlations for contrast-enhanced measurements of signal intensity were stronger than those for plain magnetic resonance imaging. Immunohistochemistry confirmed that contrast medium enhancement reflects the vascular status of the graft tissue during remodeling. We conclude that quantitatively determined magnetic resonance imaging signal intensity may be a useful tool for following the graft remodeling process in a noninvasive manner. PMID- 11734491 TI - The position of the tibia during graft fixation affects knee kinematics and graft forces for anterior cruciate ligament reconstruction. AB - Ten cadaveric knees (donor ages, 36 to 66 years) were tested at full extension, 15 degrees, 30 degrees, and 90 degrees of flexion under a 134-N anterior tibial load. In each knee, the kinematics as well as in situ force in the graft were compared when the graft was fixed with the tibia in four different positions: full knee extension while the surgeon applied a posterior tibial load (Position 1), 30 degrees of flexion with the tibia at the neutral position of the intact knee (Position 2), 30 degrees of flexion with a 67-N posterior tibial load (Position 3), and 30 degrees of flexion with a 134-N posterior tibial load (Position 4). For Positions 1 and 2, the anterior tibial translation and the in situ forces were up to 60% greater and 36% smaller, respectively, than that of the intact knee. For Position 3, knee kinematics and in situ forces were closest to those observed in the intact knee. For Position 4, anterior tibial translation was significantly decreased by up to 2 mm and the in situ force increased up to 31 N. These results suggest that the position of the tibia during graft fixation is an important consideration for the biomechanical performance of an anterior cruciate ligament-reconstructed knee. PMID- 11734492 TI - Reproducibility of anatomic tibial landmarks for anterior cruciate ligament reconstructions. AB - We evaluated the reproducibility of landmarks used for accurate anatomic placement of the tibial tunnel in anterior cruciate ligament reconstruction. Landmarks evaluated were the medial tibial eminence, the posterior cruciate ligament, the "over-the-back" position, the true posterior border of the tibia, and the posterior border of the lateral meniscus. Forty-two pairs of cadaveric knees were dissected, and anatomic measurements were made regarding the anterior cruciate ligament insertion and these various landmarks. Statistical analysis was used to confirm reproducibility and significance. Measurements based on the medial tibial eminence and posterior border of the meniscus were particularly erratic. The most reproducible anatomic landmark was the posterior cruciate ligament. The anterior border of the posterior cruciate ligament was consistently 6.7 mm posterior to the posterior border of the anterior cruciate ligament and 10.9 mm posterior to the central sagittal insertion point of the anterior cruciate ligament. The over-the-back position was consistently in contact with the anterior border of the posterior cruciate ligament if the knee was flexed with a posterior-directed force applied. In this position, the over-the-back position was equally reproducible as compared with the posterior cruciate ligament. Measurements gauged from the true posterior border of the tibia gave a second rigid bony landmark but with a wider standard deviation than the posterior cruciate ligament-based landmarks. The relative anterior-posterior dimension of the tibia did not correlate with the relationship between the anterior cruciate ligament and other anatomic landmarks. PMID- 11734493 TI - The relationship of the femoral origin of the anterior cruciate ligament and the distal femoral physeal plate in the skeletally immature knee. An anatomic study. AB - We defined the anatomic relationship of the anterior cruciate ligament femoral origin to the distal femoral physis in the skeletally immature knee with use of 12 fresh-frozen human fetal specimens (ages, 20 to 36 weeks). Each specimen underwent magnetic resonance imaging, was dissected free of soft tissue, sectioned in the sagittal plane, and stained. The spatial relationship of 1) the epiphyseal side of the physeal proliferative zone to the nearest point of bony attachment of the anterior cruciate ligament and 2) the origin of the anterior cruciate ligament to the over-the-top position were measured. The same measurements were made in 13 skeletally immature knees (ages, 5 to 15 years). We found that the femoral origin of the fetal anterior cruciate ligament developed as a confluence of ligament fibers with periosteum at 20 weeks, vascular invasion into the epiphysis at 24 weeks, and establishment of a secure epiphyseal attachment by 36 weeks. In the fetus, the distance from the anterior cruciate ligament femoral origin to the epiphysis was 2.66+/-0.18 mm (range, 2.34 to 2.94). There was no significant change in this distance in adolescent specimens (2.92+/-0.68 mm; range, 2.24 to 3.62). The over-the-top position was at the level of the distal femoral physis. PMID- 11734494 TI - Effect of arm elevation and rotation on the strain in the repaired rotator cuff tendon. A cadaveric study. AB - In 14 cadaveric shoulders, a rotator cuff tear (2 cm wide and 1.5 cm long) was created and repaired under a 3-kg tensile force with the arm in adduction. Strain on the repaired tendon was measured at 0 degrees, 15 degrees, 30 degrees, and 45 degrees of elevation in the sagittal, scapular, and coronal planes and from 60 degrees of internal rotation to 60 degrees of external rotation. The strain in all of the planes decreased significantly with the arm elevated more than 30 degrees. With 30 degrees of elevation in the scapular and coronal planes, the strain increased in internal rotation and decreased in external rotation. In all of the positions measured, the strain in the sagittal plane was significantly greater than in the other planes. We concluded that more than 30 degrees of elevation in the coronal or scapular plane and rotation ranging from 0 degrees to 60 degrees of external rotation compose the safe range of motion after repair of the rotator cuff. PMID- 11734495 TI - A history of low back injury is a risk factor for recurrent back injuries in varsity athletes. AB - In this prospective study, we investigated whether a history of previous low back injury and dissatisfaction with a coach and teammates could predict future low back injury in varsity athletes during a 1-year follow-up period. Of 679 Yale varsity athletes surveyed in 1999, 18.3% (124) reported that they had sustained a low back injury within the past 5 years, and 6.8% (46) sustained a low back injury in the follow-up season. There were no differences in incidence rates between men and women or between athletes involved in contact or noncontact sports. A history of low back injury was the significant predictor for sustaining low back injury in the following year, and athletes who reported previous low back injury were at three times greater risk. Athletes who still had pain at the time of the survey were six times more likely to sustain a low back injury than were athletes without a history of low back injury. These results suggest that some risk factors associated with a history of low back injury predispose athletes to sustain recurrent injury. They may be congenital or a result of insufficient recovery time after the first low back injury episode. PMID- 11734496 TI - A cyclooxygenase-2 inhibitor impairs ligament healing in the rat. AB - Celecoxib was the first of a new class of nonsteroidal antiinflammatory drugs, the cyclooxygenase-2 (COX-2) specific inhibitors, marketed as having the same antiinflammatory efficacy as other nonsteroidal antiinflammatory drugs without their increased risk of gastrointestinal ulceration. Among the widest uses of nonsteroidal antiinflammatory drugs is in the treatment of acute soft tissue injuries. Although the benefits of celecoxib have been shown when used for rheumatoid arthritis and osteoarthritis, we are unaware of any studies concerning its effect on soft tissues. We used the surgically incised medial collateral ligament of male Sprague-Dawley rats as an experimental model for acute ligament injuries to investigate the effects of celecoxib on ligament healing. Fifty rats underwent surgical transection of the right medial collateral ligament. Postoperatively, half were given celecoxib for the first 6 days of recovery, the other half were not. The animals were sacrificed 14 days after the operation, and both the injured and uninjured medial collateral ligaments were mechanically tested to failure in tension. Celecoxib-treated/injured ligaments were found to have a 32% lower load to failure than untreated/injured ligaments. The results of this study do not support use of cyclooxygenase-2 specific inhibitors in the treatment of ligament injuries. PMID- 11734497 TI - Quality of life assessment in elite collegiate athletes. AB - The objectives of this study were to establish baseline Medical Outcomes Study Short Form Health Survey (SF-36) data for Division I collegiate athletes and to determine the effects of injury severity and training time. All participating athletes (N = 562) at a major university were evaluated with the SF-36. Regression analysis was performed to identify predictive factors. When the men without injury were compared with a previously established norm group, there was a significant increase in the role emotional score. In the women without injury there were significant increases in mental component summary, physical function, role emotional, mental health, and vitality scores when compared with the norm group. Serious injury was a predictor of lower scores in all domains, whereas minimal injury was predictive of lower physical component summary, role physical, bodily pain, social function, and general health scores. Increased training time was predictive of higher mental component summary, role physical, vitality, and general health scores. Elite collegiate athletes scored differently from previously established age-matched norms, and injury was a strong predictor of lower scores. PMID- 11734498 TI - Bilateral femoral neck stress fractures in an adolescent male runner. A case report. PMID- 11734499 TI - Isolated femoral mononeuropathy in the athlete. Anatomic considerations and report of two cases. PMID- 11734500 TI - Quadriceps muscle contusion resulting in osteomyelitis of the femur in a high school football player. A case report. PMID- 11734501 TI - Motion loss after ligament injuries to the knee. Part II: prevention and treatment. AB - This is the second part of a two-part review on motion problems after ligament injuries to the knee. The first part, published in the September/October 2001 issue, discussed normal and abnormal knee motion, terminology, risk factors, and pathoanatomy. The purpose of this article is to review current concepts on prevention and treatment of motion problems, summarizing the recent and pertinent studies that discuss this complicated clinical problem. The first part of this article will discuss the different classification schemes that have been published on motion loss of the knee. Prevention strategies will be discussed next, followed by early recognition. Finally, a discussion of the various treatment options and published results will be presented in detail, together with the authors' nine-step systematic surgical approach to the stiff knee. PMID- 11734502 TI - The effects of patellar taping on patellofemoral incongruence. A computed tomography study. PMID- 11734503 TI - The effects of patellar taping on patellofemoral incongruence. A computed tomography study. PMID- 11734504 TI - Disability days in major league baseball. PMID- 11734505 TI - Blinding trachoma: the forgotten problem. PMID- 11734506 TI - The role of choroidal haemodynamic abnormalities in the pathogenesis of age related macular degeneration. PMID- 11734507 TI - Amniotic membrane transplantation for persistent corneal epithelial defect. PMID- 11734508 TI - Genetic testing--swings and roundabouts: a view from the United Kingdom. PMID- 11734509 TI - The burden of trachoma in the rural Nile Delta of Egypt: a survey of Menofiya governorate. AB - BACKGROUND: Evidence of widespread distribution of trachoma in Egypt had not been clarified as previous surveys were limited to individual communities which may not have been representative of the general population. The Nile Delta of Egypt presents a unique environment for trachoma to persist. Economic improvements in the past decade have affected even the poorest rural environments; availability of electricity is now found in many rural communities. Availability of water in Nile Delta has always been good but poor hygienic conditions have been the primary factor in trachoma transmission. A survey of trachoma was undertaken in Menofiya governorate to determine if Egypt should be identified as trachoma endemic and targeted for trachoma control efforts. METHODS: A multistage random cluster study design was used with the target population defined as adults aged 50 and over and children aged 2-6 years from throughout the governorate. Among preschool children only trachoma was graded while among adults presenting visual acuity and cause of vision loss or blindness were also recorded. Adults were interviewed regarding past trichiasis surgery; those currently with trichiasis or a history of trichiasis surgery were also interviewed regarding outcome of surgery. RESULTS: A total of 3272 children aged 2-6 and 3322 adults age 50+ were enumerated. Among the children 81.3% were examined and among the adults 73.0% were examined. Active trachoma (follicles (TF) and/or intense inflammation (TI)) was found among 36.5% (95% confidence interval (CI) 34.7-38.3%) of the children. TI was 1.89 (95% CI 1.22-2.94) times more common in rural children compared to urban children. The prevalence of trichiasis (TT) in adults was 6.5%; women had an age adjusted odds of trichiasis of 1.68 (95% CI 1.18-2.39) compared to men. Trichiasis was 2.11 times (95% CI 1.33-3.37) more common in rural Menofiya compared to urban Menofiya. TT accounts for blindness (presenting vision <3/60) in 8% of patients and accounts for 13.2% of visual impairment. Overall, trichiasis surgical coverage was 34.4%, slightly higher among men than women. The outcome of trichiasis surgery was poor in 44.4% of cases. CONCLUSION: Trachoma is a serious public health problem in Menofiya governorate and a significant contributor to vision loss. These findings would suggest that continued poor hygienic conditions in rural Egypt have limited the reduction of active trachoma even in the face of significant improvements in socioeconomic status. Furthermore, the high proportion of trichiasis surgery cases with a poor outcome would indicate a need to reassess current surgical practices in Egypt and improve training and monitoring. PMID- 11734510 TI - Pulsatile ocular blood flow in asymmetric exudative age related macular degeneration. AB - BACKGROUND/AIMS: Decreased perfusion or increased vascular resistance of the choroidal vessels had been proposed as the vascular pathogenesis for age related macular degeneration (AMD). This study planned to answer the question whether pulsatile ocular blood flow (POBF) was different in patients with asymmetric exudative AMD between eyes with drusen, choroidal neovascularisation (CNV), or disciform scar. METHODS: 37 patients with asymmetric exudative AMD were enrolled in this observational case series study. POBF were measured in both eyes of each subject. Eyes with high myopia, anisometropia, recent laser treatment, and glaucoma were excluded. RESULTS: After adjusting for ocular perfusion pressure, intraocular pressure, and pulse rate, multivariate regression analysis with generalised estimating equation showed POBF was significantly higher in eyes with CNV (1217 (SD 476) microl/min) than the contralateral eyes with drusen (1028 (385) microl/min) (p = 0.024). Eyes with disciform scar had lower POBF than the contralateral eyes with drusen (999 (262) microl/min and 1278 (341) microl/min, respectively, p<0.001). There was no significant correlation between the POBF and the lesion size of the CNV. CONCLUSION: The POBF in eyes with drusen was lower than their fellow eyes with CNV, but higher than their fellow eyes with disciform scar. This finding suggests that haemodynamic differences between fellow eyes in individuals are relevant to the development of CNV and the formation of disciform scar. Further studies on the follow up patients might shed light on the pathogenesis of exudative AMD. PMID- 11734511 TI - Effect of three different media on serum free culture of donor corneas and isolated human corneal endothelial cells. AB - BACKGROUND: Removal of bovine serum from organ culture medium is necessary because of the variability in serum composition and the potential risk of infection. Two specific endothelial cell media (F99 and Endothelial-SFM) were compared with the routinely used medium MEM for their use in serum free cultivation of human corneal endothelial cells (HCEC) and donor corneas. METHODS: HCEC were incubated in three test media with or without increasing serum content and a growth assay was performed. Seven pairs of donor corneas were cultured in each of three media for 3 weeks, one cornea with serum supplementation and one without. Endothelial cell density was determined once each week. Trypan blue staining of the endothelium and vital staining of keratocytes was performed after 3 weeks. RESULTS: All three media promoted proliferation of cultured HCEC when supplemented with serum. Endothelial cell density of donor corneas was comparable after 3 weeks of cultivation in the different media. Only corneas cultured in medium MEM without serum exhibited a higher endothelial cell loss. Trypan blue staining of the endothelium after cultivation revealed the lowest number of damaged cells on corneas cultured in the medium Endothelial-SFM. The highest densities of keratocytes were found in corneas cultured in Endothelial-SFM and the lowest densities occurred after culture in MEM. CONCLUSION: After incubation in Endothelial-SFM even under serum free conditions corneas were found to be of higher quality with respect to endothelial cell survival, cell membrane integrity, and keratocyte density. This medium may replace MEM, which is routinely used in European eye banks but requires supplementation with serum. PMID- 11734512 TI - Histopathological examination of two cases of anterior staphyloma associated with Peters' anomaly and persistent hyperplastic primary vitreous. AB - AIMS: To clarify the developmental mechanism and critical period for the uncommon complex of Peters' anomaly and persistent hyperplastic primary vitreous (PHPV). METHODS: Two eyes with Peters' anomaly and PHPV were histologically examined by serial section. One eye was enucleated at age 7 months (case 1) and the other at age 4 months (case 2) owing to severe anterior staphyloma. RESULTS: In both eyes, defects in the endothelium, Descemet's membrane, and posterior stroma were observed in the central cornea, and the degenerative lens adhered to the posterior surface of the defective corneal stroma. Also, in both eyes, the anterior chamber space was not formed and the undifferentiated iris stroma adhered to the posterior surface of the peripheral cornea. Mesenchymal tissue containing melanocytes was observed behind the degenerative lens, and the pigment epithelium was absent at the lower nasal side of the ciliary body in case 1. In case 2, mesenchymal tissue containing scattered melanocytes in the vitreous cavity was seen on the posterior retina. Based on the histological findings, both cases were diagnosed as Peters' anomaly caused by the faulty separation of the lens vesicle, PHPV, maldevelopment of the iris and ciliary body, and goniodysgenesis. CONCLUSION: Migratory disorders of neural crest cells from 4 to 7 weeks of gestation may be responsible for various ocular anomalies including Peters' anomaly and PHPV, as observed in these cases. PMID- 11734513 TI - Plasma malondialdehyde and nitric oxide levels in age related macular degeneration. AB - AIMS: To evaluate alteration of plasma malondialdehyde (MDA) and nitric oxide (NO) levels in patients with exudative age related macular degeneration (ARMD). METHODS: Plasma nitrite plus nitrate concentrations as an index of plasma NO levels and plasma MDA level as a marker of lipid peroxidation were measured in patients with exudative ARMD and age and sex matched healthy subjects. RESULTS: Significantly higher MDA and lower NO levels were detected in plasma of patients with ARMD compared with their controls (p=0.01, p=0.001, respectively). CONCLUSION: The results may support involvement of oxidative damage and vascular theory in the pathogenesis of ARMD as part of the ageing process. PMID- 11734514 TI - Clinical features of a novel TIMP-3 mutation causing Sorsby's fundus dystrophy: implications for disease mechanism. AB - AIMS: To describe the phenotype in three family members affected by a novel mutation in the gene coding for the enzyme tissue inhibitor of metalloproteinase 3 (TIMP-3). METHODS: Three members of the same family were seen with a history of nyctalopia and visual loss due to maculopathy. Clinical features were consistent with Sorsby's fundus dystrophy. Exon 5 of the gene coding for TIMP-3 was amplified by the polymerase chain reaction, single strand conformation polymorphism analysis undertaken and exon 5 amplicons were directly sequenced. RESULTS: Onset of symptoms was in the third to fourth decade. Five of six eyes had geographic macular atrophy rather than neovascularisation as a cause for central visual loss. Peripheral retinal pigmentary disturbances were present. Scotopic ERGs were abnormal in all three. Mutation analysis showed a G-->T transversion in all three resulting in a premature termination codon, E139X, deleting most of the carboxy terminal domain of TIMP-3. CONCLUSIONS: The patients described had a form of Sorsby's fundus dystrophy which fell at the severe end of the spectrum of this disease. Postulated disease mechanisms include deposition of dimerised TIMP-3 protein. PMID- 11734515 TI - Short wavelength automated perimetry in age related maculopathy. AB - BACKGROUND/AIMS: Previous studies reported the predictive value of the short wavelength sensitive (SWS) cone mediated sensitivity for visual outcome in age related macular degeneration. In this study SWS sensitivity was measured by commercially available blue on yellow perimetry in patients with non-exudative age related maculopathy (ARM) and compared with the presence of morphological risk factors and the status of the fellow eye. METHODS: In a prospective cross sectional study, 126 patients (57 males, 69 females, mean age 71 (SD 6) years) with ARM (visual acuity >20/50) were tested. Central visual fields (blue on yellow) were obtained with a conventional perimeter. Fundus slides were graded by two independent observers for soft drusen and presence of focal hyperpigmentation. RESULTS: Mean sensitivity and standard deviation of all patients exhibited a significant reduction with age. Patients with soft drusen had significantly lower sensitivity than those without, whereas there were no differences in visual acuity (log MAR). Sensitivity was also reduced in those eyes with fellow eyes having a sight threatening complication of age related macular degeneration (AMD). Eyes with focal hyperpigmentation compared with those without had no loss of sensitivity, but did have a significant decrease in the central part of the field compared with the more eccentric. CONCLUSION: SWS sensitivity loss is associated with common risk factors for progression to AMD. Short wavelength automated perimetry is moderately rapid and readily available. It may serve as a tool in future ARM trials. PMID- 11734516 TI - Anatomical and visual outcome of retinal detachment surgery in children. AB - AIMS: To evaluate the visual and anatomical outcome, as well as complications following surgery, for rhegmatogenous retinal detachment in children at a tertiary referral centre over a 5 year period. METHODS: A retrospective survey of all children (aged 0-16 years) who underwent primary retinal detachment surgery at Queen's Medical Centre between April 1994 and March 1999. RESULTS: 15 consecutive patients were identified with a mean follow up of 14.7 months (range 3-57) and a mean age of 12.4 years. Trauma was the cause in 40% (6/15). Complete retinal reattachment was achieved in 86.6% (13/15). Visual improvement occurred in 53.3% (8/15), worsening of vision occurred in 13.3% (2/15), with no change in the remaining 33.3% (5/15). Visual acuity was D 6/12 in 6.6% (1/15) preoperatively, and 26.6% (4/15) postoperatively. CONCLUSIONS: Retinal detachment in children is rare compared with adults. There are therefore limited data available for this group of patients. These data provide one unit's experience over a 5 year period, and may help provide a basis for information for patients and their parents when discussing the risks and potential benefits of surgery for retinal detachment in the paediatric population. PMID- 11734517 TI - Prediction of prognosis in patients with uveal melanoma using fluorescence in situ hybridisation. AB - BACKGROUND/AIMS: Uveal melanoma is the commonest malignancy of the eye, with a high proportion of patients dying of metastatic disease. Tumours showing a loss of chromosome 3 and gains of chromosome 8 are associated with a worse prognosis. The efficiency of fluorescence in situ hybridisation (FISH) in determining copy numbers of these chromosomes was assessed in individual tumours and related to patient survival. METHODS: 33 fresh frozen samples were analysed with centromeric probes for chromosomes 3 and 8. Patient outcomes were divided into two groups: (1) absence of genetic abnormalities (no genetic imbalance) and (2) presence of genetic abnormalities (genetic imbalance). The log rank test was used to compare survival, which was represented by Kaplan-Meier survival curves. RESULTS: Of the 33 tumours analysed, 16 showed evidence of genetic imbalances. Of these 16 tumours, 14 patients had died by the end of the study, with 10 having died of liver metastases. Of the tumours without evidence of genetic imbalances, five patients had died by the end of the study, although none had died as a result of either liver metastases or from the primary uveal melanoma. The difference in survival between the two groups was highly significant (p<0.0001). CONCLUSION: The authors have shown that FISH analysis for chromosome 3 and 8 is a reliable and efficient technique in the analysis of fresh frozen tumour specimens and is valuable in the prediction of prognosis in individuals with uveal melanomas. PMID- 11734518 TI - Quantification of relative afferent pupillary defects induced by posterior sub Tenon's, peribulbar, and retrobulbar anaesthetics. AB - AIMS: The effect of local anaesthetics on optic nerve function can be investigated by quantifying the relative afferent pupillary defect (RAPD). METHODS: The study compared the depth of induced RAPD following posterior sub Tenon's, retrobulbar, and peribulbar local anaesthetics using crossed polarising filters before cataract surgery (time 1 = 5 minutes), immediately after surgery (time 2 = 42 minutes (av)), and once again on the ward (time 3 = 107 minutes (av)). RESULTS: All patients developed a RAPD. There was no significant difference in the depth of RAPD between the groups at any one time period. The peribulbar group had a significantly steeper decay in RAPD from time 1 to time 2 (p = 0.014). This effect was reduced when the shorter operation time for this group was entered as a cofactor (p = 0.063). By time 3 the RAPDs for all groups had decayed similarly so that no differences could be detected. CONCLUSION: All three anaesthetic methods caused a similar level of disruption to optic nerve conduction immediately following administration and at the time of day case discharge. PMID- 11734519 TI - Elevated visual motion detection thresholds in adults with acquired ophthalmoplegia. AB - AIMS: To test the hypothesis that in patients with acquired chronic bilateral ophthalmoplegia, abnormal retinal image slippage during head movements would result in abnormal thresholds for visual perception of motion. METHODS: Five patients (two males and three females) with ophthalmoplegia were included in the study. The average age was 44 years (range 30-69 years). The aetiology of ophthalmoplegia was myasthenia gravis (MG; n=2), chronic progressive external ophthalmoplegia (CPEO; n=2), and chronic idiopathic orbital inflammation. Visual motion detection thresholds were assessed using horizontal and vertical gratings (spatial frequency) set at thresholds for visibility. The grating was then accelerated at 0.09 deg/s(2). The subject's task was to detect the drift direction of the stimulus. RESULTS: Visual motion detection thresholds were raised to a mean of 0.434 deg/s (SD 0.09) (mean normal value 0.287 deg/s (SD 0.08)) for horizontal motion; and to a mean of 0.425 deg/s (SD 0.1) (mean normal value 0.252 deg/s (SD 0.08)) for vertical motion. The difference in values for both horizontal and vertical motion detection were statistically significant when compared with age matched controls; p <0.023 for horizontal motion and p<0.07 for vertical motion (two tailed t test). CONCLUSION: Abnormally raised visual motion thresholds were found in patients with ophthalmoplegia. This may represent a centrally mediated adaptive mechanism to ignore excessive retinal slip and thus avoid oscillopsia during head movements. PMID- 11734520 TI - Spontaneous eyelid expansion after full thickness eyelid resection and direct closure. AB - BACKGROUND/AIMS: Direct closure of eyelid defects gives excellent functional results but is usually restricted to defects measuring less than a quarter of the eyelid length for fear of distorting the palpebral aperture and compromising lid function. The authors have used direct closure in larger defects. The aim of this study was to establish the effects of direct closure of full thickness eyelid margin defects under tension on the palpebral aperture dimensions. METHODS: A consecutive series of patients who had undergone one eyelid, full thickness lid resection repaired by direct closure were identified and invited to have both eyes photographed. The palpebral apertures of both eyes were measured from the photographs by a masked observer. The amount of eyelid resected was recorded from the operation notes. The unoperated palpebral aperture was used as the control. The result were analysed using a paired samples t test. RESULTS: The photographs of 18 patients were included in the analysis. The mean width of excised full thickness lid tissue was 15 mm (range 7-26 mm). The mean vertical palpebral aperture height was 9.2 (SD 1.4) mm in the operated eye as opposed to 9.3 (SD 1.2) mm in the non-operated eye. The mean horizontal palpebral aperture width was 26.1 (SD 1.9) mm in the operated eye as opposed to 26.4 (SD 1.8) mm in the non operated eye. There was no statistically significant difference between the operated and unoperated horizontal and vertical palpebral measurements. CONCLUSIONS: Direct closure of large full thickness eyelid defects is possible in selected patients with excellent functional and cosmetic results. Eyelid tissue expansion occurs spontaneously following direct eyelid defect closure under tension, restoring the palpebral aperture dimensions. PMID- 11734521 TI - Single and multilayer amniotic membrane transplantation for persistent corneal epithelial defect with and without stromal thinning and perforation. AB - AIMS: To evaluate the efficacy of amniotic membrane transplantation (AMT) in persistent corneal epithelial defect with or without stromal thinning and corneal perforation. METHODS: 28 patients (28 eyes) with persistent corneal epithelial defect unresponsive to medical treatment were given preserved human amniotic membrane transplants. The patients were divided into three groups: group A, persistent corneal epithelial defect 10 eyes; group B, epithelial defect with stromal thinning 13 eyes; and group C, corneal perforation five eyes. AMT was performed using one layer in group A and multilayers in group B and C. The causes of persistent epithelial defect were neurotrophic keratopathy (24 eyes), limbal deficiency (six eyes), exposure keratopathy (four eyes), and Mooren's ulcer (one eye). RESULTS: Success was noted in 82.1% (23/28 eyes) in all groups, with 80% (8/10 eyes), 84.6% (11/13 eyes), and 80% (4/5 eyes) in groups A, B, and C respectively, with a mean follow up of 10.9 months (1-30 months). The mean epithelialisation time after AMT was 2.1 weeks. The healing times of groups B and C are also significantly shorter than group A (p=0.017 and 0.018, respectively). Corneal stromal thickness was significantly increased in all cases in groups B and C (p=0.006). Those with corneal perforation in group C were completely healed by multilayer AMT. There was no difference in the epithelialisation time between successful cases treated by a single operation (17 eyes) or repeated operation (six eyes). Vision improved in 18.9% (8/28 eyes) and worsened as a result of cataract formation in 2.3% (1/28 eyes). Failure was noted in 17.9% (5/28 eyes), because of corneal infection (two eyes), neurotrophic keratopathy with and without limbal deficiency (two eyes), and intractable corneal perforation (one eye). No patient developed major immediate postoperative complications or graft rejection. CONCLUSION: Amniotic membrane can successfully treat refractory corneal epithelial defect by promoting epithelial healing and thus prevent corneal perforation. It can be used as a treatment for corneal perforation by restoring corneal stromal thickness so that emergency penetrating keratoplasty can be avoided. PMID- 11734522 TI - Systemic cyclosporin A in high risk penetrating keratoplasties: a case-control study. AB - AIM: To examine the efficacy of systemic cyclosporin A (CSA) in preventing rejection and graft failure in high risk keratoplasty (PK). METHODS: A retrospective case-control study with 49 patients in both the CSA group and the control group. The patients receiving CSA were at high risk of graft rejection and failure. Controls were identified from surgical audit books and had high risk characteristics. RESULTS: There was no statistical difference in preoperative risk factors and the use of postoperative topical steroids between the two groups. The median follow up in the CSA group was 22 months and 27 months in the control group. One or more rejection episodes occurred in 18 out of 49 (36.7%) cases in the CSA group and 26 out of 49 (53.1%) in the control group. Graft failure from all causes occurred in 16 (32.7%) CSA patients and 18 (36.7%) control patients. Four (8.2%) of the CSA group compared to eight (16.3%) in the control group failed because of rejection. 22 (44.9%) out of 49 patients in the CSA group had side effects. In five (10.2%) patients, CSA was stopped because of the side effects; eight patients had elevated serum urea and creatinine and four developed hypertension. Minor side effects reported include gum hyperplasia, increased sweating, backache, nausea, feeling unwell, oral candidiasis, cramps, and paraesthesia of the extremities. CONCLUSION: These results suggest that the benefit of CSA over conventional therapy in preventing rejection episodes and subsequent graft failure is only moderate and did not reach statistically significant levels in this study. Considering the high frequency of side effects and the cost of CSA, a randomised control trial may be necessary to determine the true value of CSA in high risk penetrating keratoplasty. PMID- 11734523 TI - The importance of genes and environment for ocular refraction and its determiners: a population based study among 20-45 year old twins. AB - AIMS: To estimate the heritability for ocular refraction and its determiners in a population based cohort of 20-45 years old twins. METHODS: 114 twin pairs (53 monozygotic and 61 dizygotic) participated. Refraction was determined in cycloplegia and eye dimensions were measured with ultrasound. Educational length was assessed. The heritability was estimated employing aetiological model fitting. Evidence of gene-environment interaction was analysed. Correlations between intrapairwise differences in educational length and in refraction were evaluated. RESULTS: The heritability was between 0.89 and 0.94 (95% CI: 0.82, 0.96) for refraction, total refraction, axial length, and radius of corneal curvature. Phenotypic variation was mostly due to additive genetic effects. Refraction revealed evidence of gene-environment interaction (r = -0.29 to -0.32; p <0.05). The heritability for anterior chamber depth and lens thickness was between 0.88 and 0.94 (95% CI: 0.81, 0.96) and dominant genetic effects were the most likely explanation. There was no correlation between age and intrapairwise differences in refraction. The dizygotic twins had significant larger intrapairwise differences in educational length (p <0.05), but the differences were not correlated with differences in refraction. CONCLUSIONS: The results indicate a high heritability for ocular refraction and its determiners and thus suggest that environmental impact on refraction is not significant. However, the epidemiological association between educational length (near work) and myopia, the evidence of increasing myopia prevalence within a few generations, and the theory of gene-environment interaction imply that some individuals might be genetically liable to develop myopia if exposed to certain environmental factors. PMID- 11734524 TI - Comparison of the main sequence of reflexive saccades and the quick phases of optokinetic nystagmus. AB - BACKGROUND/AIMS: Abnormalities in the saccadic main sequence are an important finding and may indicate pathology of the ocular motor periphery or central neurological disorders. In young or uncooperative patients it can be difficult eliciting a sufficient number of saccades to measure the main sequence. It is often assumed that the quick phases of optokinetic nystagmus (OKN) are identical to saccades. If this were the case, it would be feasible to use OKN, an involuntary response that is easily evoked, as a simple way of eliciting many saccades. The aim of this study was to determine whether reflexive saccades and the quick phases of OKN are indeed identical, and whether OKN quick phases could have a clinical role in identifying patients with slow saccades. METHODS: OKN and reflexive saccades were recorded from 10 healthy adults using an infrared limbus eye tracker and bitemporal DC electro-oculography simultaneously. OKN was stimulated by rotating a full field patterned curtain around the subject at 10-50 degrees /s. Reflexive saccades were elicited to red LED targets at 5-20 degrees eccentricity. RESULTS: OKN quick phases tended to have a longer duration compared to saccades, but these differences were not significant. OKN quick phases had a slightly lower peak velocity compared to saccades, which was statistically significant (p<0.05). CONCLUSION: The main sequence for duration is the same for reflexive saccades and OKN quick phases. The main sequence for peak velocity is slightly faster for reflexive saccades than OKN quick phases, but the difference is unlikely to be of clinical significance. As an illustration of the potential of this technique, the authors demonstrate that OKN quick phases show similar slowness to saccades in a child with brainstem pathology caused by Gaucher disease type III. It is concluded that recording OKN may be a simple clinical means for approximating the main sequence. PMID- 11734525 TI - Socioeconomic status and blindness. PMID- 11734527 TI - Analysis of publication trends in two internationally renowned ophthalmology journals. AB - International journals represent a forum for exchange of current information with contributions from all over the world. High standards are essential. In this report, we compared the publishing trends of two internationally renowned ophthalmology journals--the British Journal of Ophthalmology (BJO) and the American Journal of Ophthalmology (AJO). PMID- 11734526 TI - Is it time for a new attitude to "simultaneous" bilateral cataract surgery? PMID- 11734528 TI - Electrophysiological properties of pyramidal neurons in the rat prefrontal cortex: an in vivo intracellular recording study. AB - In order to determine the electrophysiological properties of prefrontal cortex pyramidal neurons in vivo, intracellular recordings coupled with neurobiotin injection were performed in anesthetized rats. Three main classes of pyramidal cells were distinguished according to both their firing patterns in response to depolarizing current pulses and the characteristics of their action potentials: regular spiking (RS, n = 71); intrinsic (inactivating) bursting (IB, n = 8); and non-inactivating bursting (NIB, n = 26) cells. RS cells were further subdivided into slow-adapting and fast-adapting types, according to their firing frequency adaptation. IB and fast-adapting RS cells could exhibit different firing patterns depending on the intensity of the depolarizing current. In response to successive depolarizing pulses of a given intensity, NIB and some RS cells showed variations in their firing patterns, probably due to the impact of local synaptic activity. All the labeled neurons were pyramidal cells with an apical dendrite that formed a terminal tuft in layer I. As compared to RS cells, NIB cells had a smaller somatic size and their apical dendritic tuft was less extensive, while IB cells presented a larger somatic size, thicker dendrites and a wider extent of their basal and apical dendritic arborization. In conclusion, we found in the rat prefrontal cortex, in vivo, different electrophysiological classes of pyramidal cells whose output firing patterns depend on interactions between their intrinsic properties and the ongoing synaptic activity. PMID- 11734529 TI - Mapping sulcal pattern asymmetry and local cortical surface gray matter distribution in vivo: maturation in perisylvian cortices. AB - Previous in vivo morphometric studies of human brain maturation between childhood and young adulthood have revealed a spatial and temporal pattern of progressive brain changes that is consistent with the post mortem cytoarchitectonic and cognitive developmental literatures. In this study, we mapped age differences in structural asymmetries at the cortical surface in groups of normally developing children (7-11 years), adolescents (12-16 years) and young adults (23-30 years) using novel surface-based mesh modeling image analytic methods. We also assessed relationships between cortical surface sulcal asymmetry and the local density of the underlying cortical gray matter. Results from this study reveal that perisylvian sulcal asymmetries are much more prominent in the adults than in the children studied. The superior posterior extent of the Sylvian fissure in the right hemisphere is approximately 7 mm more superior in the average adult than in the average child studied, whereas little difference is observed during this age range in the location of this anatomical structure in the left hemisphere. Age related differences in Sylvian fissure asymmetry were significant (P = 0.0129, permutation test), showing increased asymmetry with increasing age. We also show age-related increases in local gray matter proportion bilaterally in the temporo parietal cortices that are anatomically and temporally related to the sulcal asymmetries. Results from this cross-sectional study imply that asymmetries in the Sylvian fissure are dynamically changing into young adulthood and show that variability in brain tissue density is related to asymmetry in this region. These morphological differences may be related to changing cognitive abilities and are relevant in interpreting results from studies of abnormal brain development where perisylvian brain regions are implicated. PMID- 11734530 TI - The effects of age on the cells in layer 1 of primate cerebral cortex. AB - Although there is significant thinning of layer 1 with age in both occipital area 17 and prefrontal area 46 of the rhesus monkey, there are no significant age related changes in the numbers of neurons, astrocytes, or microglia and oligodendrocytes in this layer. A few profiles of degenerating neurons have been encountered in old monkeys, but they are uncommon. Some astrocytes undergo hypertrophy with age, as evidenced by the increased thickness of the glial limiting membrane, and throughout layer 1 the amount of filaments in the cytoplasm of both their cell bodies and processes increases. The astrocytes also come to contain phagocytic material in the old monkeys, as do the microglial cells. We have previously shown that in both areas 17 and 46 there is an age related loss of synapses from layer 1 and a concomitant loss of dendritic branches from the apical tufts of pyramidal cells from layer 1. These may be the sources of the material phagocytosed by the astrocytes and microglial cells. PMID- 11734532 TI - Early cortical distinction between memories that pertain to ongoing reality and memories that don't. AB - Patients with anterior limbic lesions, in particular of the posterior orbitofrontal cortex, often act on the basis of memories that do not relate to ongoing reality and justify their behavior with invented stories that can mostly be traced back to real events (spontaneous confabulation). Recent studies demonstrated that the patients fail to suppress activated memory traces that do not pertain to ongoing reality. In the present study, we used a similar paradigm and high-resolution event-related potentials to explore when this suppression happens. Healthy subjects made two runs of a continuous recognition task, composed of the same set of pictures, and were requested to indicate picture recurrences only within the ongoing run. Thus, performance in the first run depends on new learning, whereas the second run requires the ability to realize whether a picture is solely familiar from its occurrence in the previous run ('distracter') or whether it has already appeared in the ongoing second run ('target'). We find that correct rejection (suppression) of currently irrelevant pictures (distracters of run 2) is associated with absent negative deflection of a frontal potential and absence of a specific cortical potential map configuration after 220-300 ms. By contrast, learning and recognition of repeatedly presented information is associated with cortical amplitude modulation after 400-480 ms. These findings indicate that by the time the content of a mental association is recognized and consolidated, its cortical representation has already been adjusted according to whether it relates to ongoing reality or not. This sequence may also explain the ability to distinguish between the memory of a true event and the memory of a thought. PMID- 11734533 TI - Contribution of GABAergic inhibition to receptive field structures of monkey inferior temporal neurons. AB - Receptive field (RF) structures of neurons in area TE of the monkey inferior temporal cortex were investigated under blockade of inhibition mediated by gamma aminobutyric acid (GABA). Bicuculline methiodide, a GABA(A) receptor antagonist, was microiontophoretically administered to TE neurons. Blockade of inhibition enhanced responses to a particular range of visual stimuli not only at the RF center, but also at the periphery of or outside RFs where the stimuli originally evoked little or no response, enlarging the RFs. The strongest responses under normal and uninhibited conditions occurred at the RF center in most neurons. The largest increase in responses, reflecting the strongest inhibitory input, usually occurred at the RF center, but in some neurons it occurred in the periphery. A neuron had a silent region within its RF where some stimuli effective at adjacent locations could not elicit responses even under blockade of inhibition. We suggest that (i) afferent information to individual TE neurons originates from a wide retinotopic region beyond their normal RF; (ii) the afferent convergence is not necessarily complete throughout a RF; and (iii) GABAergic inhibition contributes to the generation of RF structures of TE neurons. PMID- 11734531 TI - Unique morphological features of the proliferative zones and postmitotic compartments of the neural epithelium giving rise to striate and extrastriate cortex in the monkey. AB - We examined the development of the occipital lobe in fetal monkeys between embryonic day 37 (E37) and E108 in Nissl-stained and acetylcholine esterase (AChE)-reacted sections. We paid particular attention to features that distinguish the development of presumptive area 17. At E46 the neuroepithelium consists of a ventricular zone and a monolayer cortical plate sandwiched between a thin marginal zone and a minimal presubplate. Between E55 and E65 an augmented subplate emerges and continues to expand up to E94 to become a major compartment of the developing cortex. A mitotic subventricular zone is established by E55. Peaking in depth at E72, it constitutes the principal germinal zone. By E78 an invading fibre tract divides it into an outer radially organized zone and a more conventional inner zone. AChE staining reveals the future area 17/18 border from E86 onwards. Proceeding from presumptive area 17 to area 18 there is a progressive thinning of the radially structured subventricular zone. Comparison of these results with corticogenesis in rodents suggests a number of potentially unique primate features: (i) a minimal preplate stage; (ii) a radially augmented germinal zone not previously described in non-primates; (iii) a fibre tract dividing the subventricular zone into two laminae; (iv) late generation and expansion of the subplate. PMID- 11734534 TI - Expression from a Dlx gene enhancer marks adult mouse cortical GABAergic neurons. AB - In this paper we analyse the expression pattern of a zebrafish dlx4/6 enhancer/reporter construct in embryonic transgenic mice. We show that the pattern of LacZ/beta-galactosidase in cells that tangentially migrate from the ganglionic eminences to the cerebral cortex is identical to that of various subpallial markers, namely Dlx and GAD genes, that are known to label this population. Because beta-galactosidase activity persists long after expression of the Dlx genes and the transgene becomes undetectable, we were able to analyse the beta-galactosidase-positive cell population of the mature cortex through X-gal staining and immunohistochemistry. We show that this population is largely identical with the adult cortical and hippocampal interneuron population, providing further evidence for their subpallial origin. PMID- 11734535 TI - Integration of bilateral whisker stimuli in rats: role of the whisker barrel cortices. AB - Recently, we demonstrated that neural responses within the whisker region of the primary somatosensory cortex (SIw) of rats are profoundly influenced by the spatiotemporal attributes of ipsilateral, as well as contralateral, whisker stimuli. As inactivation of one SIw eliminates in the intact SIw both ipsilaterally evoked responses and the influence of ipsilateral stimulation on contralaterally evoked activity, we proposed that interhemispheric interactions between the SIws may be important for integrating bilateral whisker information. To test whether rats can recognize the bilateral nature of a whisker stimulus, we developed a tactile discrimination task that required rats to conjointly determine distances to a left and a right discriminandum as equidistant or non equidistant using only their facial whiskers. All rats trained in this task achieved performance levels indicative of an ability to integrate bilateral whisker information. Testing during unilateral, as well as bilateral, inactivation of the SIws indicated that rats rely on both SIws for detecting the bilateral nature of a whisker stimulus. Rats were unable to perform the task without both sets of whiskers, a fact that indicates that the whiskers (and not other modalities) were used to perform this task. The findings presented here indicate that rats can solve a task that requires the conjoint detection of left and right whisker-mediated distance information and implicate the SIws as central to this ability. PMID- 11734536 TI - Motor map expansion following repeated cortical and limbic seizures is related to synaptic potentiation. AB - The effect of experimentally induced seizure activity on the functional reorganization of motor maps has not previously been investigated. Furthermore, while the functional reorganization of motor maps has been thought to involve increases in synaptic communication, there has yet to be a test of this hypothesis. Here we show that repeated seizure activity (kindling), that is accompanied by increased synaptic strength within adult rat motor cortex, results in a doubling of the caudal forelimb motor area. We measured neo-cortical evoked potentials in the right hemisphere prior to 25 days of electrical kindling of the medial frontal corpus callosum or amygdala and re-measured them either 1 or 21 days following the last kindling session. Then, using high resolution intracortical microstimulation (ICMS), the caudal forelimb area in the left hemisphere was mapped. This is the first report of any procedure causing a motor representation to double in size. Furthermore, this expansion was related to the enhanced area of a neocortical polysynaptic field potential and not the motor convulsions per se. Moreover, both the motor map and field potential enhancements were persistent in nature and could be driven from either cortical or limbic sites. The data have implications for human populations with epilepsy. PMID- 11734538 TI - A functional assay for mutations in tumor suppressor genes caused by mismatch repair deficiency. AB - The coding sequences of multiple human tumor suppressor genes include microsatellite sequences that are prone to mutations. Saccharomyces cerevisiae strains deficient in DNA mismatch repair (MMR) can be used to determine de novo mutation rates of these human tumor suppressor genes as well as any other gene sequence. Microsatellites in human TGFBR2, PTEN and APC genes were placed in yeast vectors and analyzed in isogenic yeast strains that were wild-type or deletion mutants for MSH2 or MLH1. In MMR-deficient strains, the vector containing the (A)(10) microsatellite sequence of TGFBR2 had a mutation rate (mutations/cell division) of 1.4 x 10(-4), compared to a mutation rate of 1.7 x 10(-6) in the wild-type strain. In MMR-deficient strains, mutation rates in PTEN and APC were also elevated above background levels. PTEN mutation rates were higher in both msh2 (4.4 x 10-5) and mlh1 strains (2.3 x 10-5). APC mutation rates in the msh2 strain (2.4 x 10-6) and the mlh1 strain (1.7 x 10-6) were also significantly, but less dramatically, elevated over background. Mutations selected for in the yeast screen were identical to those previously observed in human tumor samples with microsatellite instability (MSI). This functional assay has applicability in providing quantitative data about microsatellite mutation rates caused by MMR deficiency in any human tumor suppressor gene sequence. It can also be applied as a genetic screen to identify new genes that are vulnerable to such microsatellite mutations and thus may be involved in the neoplastic development of tumors with MSI. PMID- 11734539 TI - Suppression of revertant fibers in mdx mice by expression of a functional dystrophin. AB - Duchenne muscular dystrophy (DMD) is characterized by progressive muscle degeneration that results from the absence of dystrophin. Despite null mutations in the dystrophin gene, many DMD patients display a low percentage of dystrophin positive fibers. These "revertant fibers" are also present in the dystrophin deficient mdx mouse and are believed to result from alternative splicing or second mutation events that bypass the mutation and restore an open reading frame. However, it is unclear what role dystrophin and the dystrophic pathology might play in revertant fiber formation and accumulation. We have analyzed the role of dystrophin expression and the dystrophic pathology in this process by monitoring revertant fibers in transgenic mdx mice that express truncated dystrophins. We found that newborn transgenic mice displayed approximately the same number of revertant fibers as newborn mdx mice, indicating that expression of a functional dystrophin does not suppress the initiation of revertant fiber formation. Surprisingly, when the transgene encoded a functional dystrophin, revertant fibers were not detected in adult or old mdx mice. In contrast, adult transgenic mice expressing a non-functional dystrophin accumulated increasing numbers of revertant fibers, similar to mdx mice, suggesting that positive selection is required for the persistence of revertant fibers. Finally, we provide evidence that the loss of revertant dystrophin in transgenic mdx muscle fibers overexpressing a functional dystrophin results from displacement of the revertant protein by the transgene-encoded dystrophin. PMID- 11734540 TI - A genome-wide scan for coronary heart disease suggests in Indo-Mauritians a susceptibility locus on chromosome 16p13 and replicates linkage with the metabolic syndrome on 3q27. AB - Prevalence of coronary heart disease (CHD), of type 2 diabetes (T2DM) and of the metabolic syndrome are in Mauritius amongst the highest in the world. As T2DM and CHD are closely associated and have both a polygenic basis, we conducted a 10 cM genome scan with 403 microsatellite markers in 99 independent families of North Eastern Indian origin including 535 individuals. Families were ascertained through a proband with CHD before 52 years of age and additional sibs with myocardial infarction (MI) or T2DM. Model-free two-point and multipoint linkage analysis were performed using the Mapmarker-Sibs (MLS) and maximum-likelihood binomial (MLB) programs for autosomal markers and the Aspex program for chromosome X markers. In a second step, additional markers were studied to increase the genetic map density in three regions on chromosomes 3, 8 and 16 where initial indication for linkage was found. Our data show suggestive linkage with CHD on chromosome 16p13-pter with the MLS statistics at 8.69 cM (LOD = 3.06, P = 0.00017) which partially overlaps with a high pressure (HBP) peak. At the same locus, a nominal indication for linkage with T2DM was found in 35 large T2DM Pondicherian families also having Indian origin. With respect to region 8q23, we found suggestive linkage with T2DM (LOD = 2.55, P = 0.00058) as well as with HBP. On 3q27, we replicated previous indication for linkage found in Caucasians (for the metabolic syndrome and for diabetes) according to the categorized trait for CHD and MI with the MLB statistics (LOD = 2.13, P = 0.0009). The genome scan also revealed nominal evidence of linkage with CHD on 10q23 (LOD = 2.06, P = 0.00188). Interestingly, we detected in the same region overlapping linkages with three QTLs: age of onset of CHD (LOD = 2.03), HDL cholesterol (LOD = 1.48) and LDL/HDL ratio (LOD = 1.34). Ordered-subset analysis based on family body mass index ranking replicated finding on 2q37 for T2DM (at Calpain 10 locus). These results show the first evidence for susceptibility loci that predispose to CHD, T2DM and HBP in the context of the metabolic syndrome. PMID- 11734541 TI - CRB1 has a cytoplasmic domain that is functionally conserved between human and Drosophila. AB - Mutations in the human Crumbs homologue 1 (CRB1) gene cause severe retinal dystrophies, ranging from retinitis pigmentosa to Leber congenital amaurosis. The CRB1 gene is expressed specifically in human retina and brain and encodes a protein homologous to the Drosophila Crumbs protein. In crumbs mutant embryos apico-basal polarity of epithelial cells is lost, leading to widespread epidermal cell death. The small cytoplasmic domain of Crumbs organizes an intracellular protein scaffold that defines the assembly of a continuous zonula adherens. The crumbs mutant phenotype can be partially rescued by expression of just the membrane-bound cytoplasmic domain, and overexpression of this domain in a wild type background results in a multilayered epidermis. A striking difference between CRB1 and Crumbs was that the latter contains a transmembrane region and a 37 amino acid cytoplasmic domain. Here we describe an alternative splice variant of human CRB1 that encodes a cytoplasmic domain 72% similar to that of Drosophila Crumbs. Two intracellular subdomains that are necessary for function in Drosophila are absolutely conserved. Rescuing and overexpression studies in Drosophila show that the cytoplasmic domains are functionally related between these distant species. This suggests that CRB1 organizes an intracellular protein scaffold in the human retina. Human homologues of proteins binding to Crumbs may be part of this complex and represent candidate genes for retinal dystrophies. PMID- 11734542 TI - Molecular effects of Eya1 domain mutations causing organ defects in BOR syndrome. AB - Eya1 is a critical gene for mammalian organogenesis. Mutations in human EYA1 cause branchio-oto-renal (BOR) syndrome, an autosomal dominant disorder characterized by varying combinations of branchial, otic and renal anomalies, whereas deletion of mouse Eya1 results in the absence of multiple organ formation. Eya1 and other Eya gene products share a highly conserved 271 amino acid Eya domain that is required for protein-protein interaction. Recently, several point mutations that result in single amino acid substitutions in the conserved Eya domain region of EYA1 have been identified in BOR patients; however, the molecular and developmental basis of organ defects that occurred in BOR syndrome is unclear. To understand how these point mutations cause disease, we have analyzed the functional importance of these Eya domain missense mutations with respect to protein complex formation and cellular localization. We have demonstrated that these point mutations do not alter protein localization. However, four mutations are crucial for protein-protein interactions in both yeast and mammalian cells. Our results provide insights into the molecular mechanisms of organ defects detected in human syndromes. PMID- 11734543 TI - Human Connexin 32, a gap junction protein altered in the X-linked form of Charcot Marie-Tooth disease, is directly regulated by the transcription factor SOX10. AB - Mutations in SOX10, a transcription modulator crucial in the development of the enteric nervous system (ENS), melanocytes and glial cells, are found in Shah Waardenburg syndrome (WS4), a neurocristopathy that associates intestinal aganglionosis, pigmentation defects and sensorineural deafness. Expression of MITF and RET, two genes that play important roles during melanocyte and ENS development, respectively, are controlled by SOX10. The observation that some WS4 patients present with myelination defects of the central and peripheral nervous systems correlates with the recent finding that P(0), a major component of the peripheral myelin, is another transcriptional target of SOX10. These phenotypic features suggest that SOX10 could regulate expression of other genes involved in the myelination process as well. Thus, we tested the ability of SOX10 to regulate expression of MBP, PMP22 and Connexin 32, three major proteins of the peripheral myelin. Our study shows that this factor, in synergy with EGR2, strongly activates Cx32 expression in vitro by directly binding to its promoter. In agreement with this finding, SOX10 and EGR2 mutants identified in patients with peripheral myelin defects fail to transactivate the Cx32 promoter. Moreover, we show that a mutation of the Cx32 promoter previously described in a patient with the X-linked form of Charcot-Marie-Tooth (CMTX) disease impairs SOX10 function. In addition to providing new insights into the molecular mechanisms underlying some of the peripheral myelin defects observed in CMTX disease, these results further extend the spectrum of genes that are regulated by SOX10. PMID- 11734544 TI - Mutations in the general transcription factor TFIIH result in beta-thalassaemia in individuals with trichothiodystrophy. AB - The transcription factor TFIIH is involved in both basal transcription and DNA repair. Mutations in the XPD helicase component of TFIIH can result in the diverse clinical features associated with xeroderma pigmentosum (XP) and trichothiodystrophy (TTD). It is generally believed that the multi-system abnormalities associated with TTD are the result of a subtle deficiency in basal transcription. However, to date, there has been no clear demonstration of a defect in expression of any specific gene in individuals with these syndromes. Here we show that the specific mutations in XPD that cause TTD result in reduced expression of the beta-globin genes in these individuals. Eleven TTD patients with characterized mutations in the XPD gene have the haematological features of beta-thalassaemia trait, and reduced levels of beta-globin synthesis and beta globin mRNA. All these parameters were normal in three patients with XP. These findings provide the first evidence for reduced expression of a specific gene in TTD. They support the hypothesis that many of the clinical features of TTD result from inadequate expression of a diverse set of highly expressed genes. PMID- 11734545 TI - Developmental expression of the fragile X-related 1 proteins in mouse testis: association with microtubule elements. AB - Fragile X mental retardation 1 protein (FMRP) is the archetype of a class of cytoplasmic mRNA-binding proteins that includes the fragile X-related 1 and 2 proteins (FXR1P and FXR2P). Whereas absence of FMRP is the cause of fragile X syndrome, it is not known if FXR1P and FXR2P are associated with any pathology. It is also still elusive whether these homologous proteins can partially compensate for the absence of FMRP in the case of the fragile X syndrome. FXR1 is widely expressed in mammals and its expression pattern is complex since several mRNA variants and protein isoforms are detected. In mouse, we observed that the highest level of FXR1 is found in the adult testis. This tissue is an exception, since all known FXR1P isoforms, some of which have been considered as tissue specific, are detected in it. In young animals, changes in mRNA-spliced variants and their corresponding protein isoforms occur during spermatogenesis. Using biochemical, immunohistochemical and electron microscopic techniques, we show that FXR1P is associated with microtubule elements. Since the cytoskeletal framework is implicated in cellular plasticity as well as in mRNA transport, we propose new possibilities for the function(s) of the FXR proteins. PMID- 11734546 TI - A new sequence motif linking lissencephaly, Treacher Collins and oral-facial digital type 1 syndromes, microtubule dynamics and cell migration. AB - A previously unidentified sequence motif has been identified in the products of genes mutated in Miller-Dieker lissencephaly, Treacher Collins, oral-facial digital type 1 and contiguous syndrome ocular albinism with late onset sensorineural deafness syndromes. An additional homologous motif was detected in a gene product fused to the fibroblast growth factor receptor type 1 in patients with an atypical stem cell myeloproliferative disorder. In total, over 100 eukaryotic intracellular proteins are shown to possess a LIS1 homology (LisH) motif, including several katanin p60 subunits, muskelin, tonneau, LEUNIG, Nopp140, aimless and numerous WD repeat-containing beta-propeller proteins. It is suggested that LisH motifs contribute to the regulation of microtubule dynamics, either by mediating dimerization, or else by binding cytoplasmic dynein heavy chain or microtubules directly. The predicted secondary structure of LisH motifs, and their occurrence in homologues of Gbeta beta-propeller subunits, suggests that they are analogues of Ggamma subunits, and might associate with the periphery of beta-propeller domains. The finding of LisH motifs in both treacle and Nopp140 reinforces previous observations of functional similarities between these nucleolar proteins. Uncharacterized LisH motif-containing proteins represent candidates for other diseases associated with aberrant microtubule dynamics and defects of cell migration, nucleokinesis or chromosome segregation. PMID- 11734548 TI - Long-range sequence composition mirrors linkage disequilibrium pattern in a 1.13 Mb region of human chromosome 22. AB - Association studies, the most powerful tool for the identification of genes underlying complex traits, depend on the observation of linkage disequilibrium (LD) between marker alleles and the trait. The LD pattern of the human genome which determines the regional density of required markers is non-uniform, with regions of long-range LD over several hundred kilobases and regions where LD extends only over a few kilobases. Studying LD in the NF1 gene region we encountered a transition from long-range to short-range LD which coincides with a switch in the isochore pattern. This observation prompted us to investigate the regional variation in the extent of LD more systematically and we selected an isochore transition within the MN1/PITPNB gene region on chromosome 22q12.1. Long range LD characterizes the GC-poor (40% GC) parts of the sequences. No LD can be observed between closely spaced markers throughout the whole range of the GC-rich (50% GC) parts. In both cases, the NF1 and the MN1/PITPNB gene region, a clear cut transition of the long-range GC content precisely coincides with a change in the extent of observable LD. The results can be explained by a 72-fold lower recombination frequency in the GC-poor, compared to the GC-rich isochores. Although recombination is not the only factor governing LD, our findings can help to predict levels of LD and marker densities required for association studies on the basis of regional GC content. PMID- 11734547 TI - Association of ataxin-7 with the proteasome subunit S4 of the 19S regulatory complex. AB - Spinocerebellar ataxia type 7 (SCA7) is a neurodegenerative disorder characterized by ataxia and selective neuronal cell loss caused by the expansion of a translated CAG repeat encoding a polyglutamine tract in ataxin-7, the SCA7 gene product. To gain insight into ataxin-7 function and to decipher the molecular mechanisms of neurodegeneration in SCA7, a two-hybrid assay was performed to identify ataxin-7 interacting proteins. Herein, we show that ataxin 7 interacts with the ATPase subunit S4 of the proteasomal 19S regulatory complex. The ataxin-7/S4 association is modulated by the length of the polyglutamine tract whereby S4 shows a stronger association with the wild-type allele of ataxin-7. We demonstrate that endogenous ataxin-7 localizes to discrete nuclear foci that also contain additional components of the proteasomal complex. Immunohistochemical analyses suggest alterations either of the distribution or the levels of S4 immunoreactivity in neurons that degenerate in SCA7 brains. Immunoblot analyses demonstrate reduced levels of S4 in SCA7 cerebella without evident alterations in the levels of other proteasome subunits. These results suggest a role for S4 and ubiquitin-mediated proteasomal proteolysis in the molecular pathogenesis of SCA7. PMID- 11734549 TI - Aclarubicin treatment restores SMN levels to cells derived from type I spinal muscular atrophy patients. AB - Proximal spinal muscular atrophy (SMA) is a common motor neuron disorder caused by mutation of the telomeric survival of motor neuron gene SMN1. The centromeric survival of motor neuron SMN2 gene is retained in all SMA patients but does not produce sufficient SMN protein to prevent the development of clinical symptoms. The SMN1 and SMN2 genes differ functionally by a single nucleotide change. This change affects the efficiency with which exon 7 is incorporated into the mRNA transcript. Thus, SMN2 produces less full-length mRNA and protein than SMN1. We have screened a library of compounds in order to identify ones that can alter the splicing pattern of the SMN2 gene. Here, we report that the compound aclarubicin increases the retention of exon 7 into the SMN2 transcript. We show that aclarubicin effectively induces incorporation of exon 7 into SMN2 transcripts from the endogenous gene in type I SMA fibroblasts as well as into transcripts from a SMN2 minigene in the motor neuron cell line NSC34. In type I fibroblasts, treatment resulted in an increase in SMN protein and gems to normal levels. Our results suggest that alteration of splicing pattern represents a new approach to modification of gene expression in disease treatment and demonstrate the feasibility of high throughput screens to detect compounds that affect the splicing pattern of a gene. PMID- 11734550 TI - IQGAP1-mediated stimulation of transcriptional co-activation by beta-catenin is modulated by calmodulin. AB - Beta-catenin, an oncoprotein integral to cell-cell adhesion and proliferative signaling, is increased in several malignancies. The recently discovered calmodulin-binding protein IQGAP1 binds stoichiometrically to beta-catenin and regulates the association of beta-catenin with the cell-cell adhesion complex. In the present work, we investigated the role of IQGAP1 on the transcriptional co activator function of beta-catenin, and whether calmodulin modulated the functional interaction between IQGAP1 and beta-catenin. In vitro competition assays revealed that both Ca(2+)/calmodulin and apo-calmodulin, when pre-bound to IQGAP1, prevented binding of beta-catenin to IQGAP1, and partially displaced beta catenin pre-bound to IQGAP1 when added subsequently. Conversely, beta-catenin partially displaced apo-calmodulin, but not Ca(2+)/calmodulin, from IQGAP1. Overexpression of IQGAP1 in SW480 colon carcinoma cells enhanced beta-catenin mediated transcriptional co-activation by 1.72-fold, and this stimulation was significantly attenuated upon antagonism of calmodulin using the cell-permeable antagonist CGS9343B. Moreover, an IQGAP1 mutant that does not bind calmodulin was unable to stimulate beta-catenin transcriptional function. Results of pulse-chase analyses suggested that IQGAP1 slowed the turnover of soluble, but not total, beta-catenin. Immunocytochemistry revealed that IQGAP1 overexpression increased the amount of beta-catenin located in the nucleus, whereas incubation of cells with CGS9343B blocked this accumulation. Together, our results imply that IQGAP1 enhances the function of beta-catenin in the nucleus and that calmodulin regulates this stimulation. PMID- 11734551 TI - Biochemical characterization of the human copper transporter Ctr1. AB - The trace metal copper is an essential cofactor for a number of biological processes including mitochondrial oxidative phosphorylation, free radical detoxification, neurotransmitter synthesis and maturation, and iron metabolism. Consequently, copper transport at the cell surface and the delivery of copper to intracellular proteins are critical events in normal physiology. Little is known about the molecules and biochemical mechanisms responsible for copper uptake at the plasma membrane in mammals. Here, we demonstrate that human Ctr1 (hCtr1) is a component of the copper transport machinery at the plasma membrane. hCtr1 transports copper with high affinity in a time-dependent and saturable manner and is metal-specific. hCtr1-mediated (64)Cu transport is an energy-independent process and is stimulated by extracellular acidic pH and high K(+) concentrations. hCtr1 exists as a homomultimer at the plasma membrane in mammalian cells. This is the first report on the biochemical characterization of the human copper transporter hCtr1, which is important for understanding mechanisms for mammalian copper transport at the plasma membrane. PMID- 11734552 TI - Retinal neuron activity of ETS domain-binding sites in a nicotinic acetylcholine receptor gene cluster enhancer. AB - Nicotinic acetylcholine receptors (nAchRs) mediate amacrine to ganglion cell synaptic transmission in the developing mammalian retina. The clustered neuronal nAchRs subunit genes, alpha 3 and beta 4, are expressed in amacrine and ganglion cells where they are used to assemble functional receptor subtypes. The transcriptional mechanisms underlying expression of these subunits in retina are not yet known but may involve enhancers that are selectively active in retinal neurons. We previously identified a neuron-selective enhancer, beta 43', whose activity in neural cell lines is dependent on ETS domain-binding sites. To determine whether beta 43' is active in retinal neurons that express the alpha 3 and beta 4 genes, we investigated beta 43' activity in primary dissociated rat retinal cultures. We found that beta 43' is selectively active in retinal neurons compared with retinal non-neuronal cells. This activity was derived primarily from amacrine and ganglion neurons, which are the retinal neuron cell types that express the clustered genes. Moreover, beta 43' was selectively active in retinal neurons compared with cerebral cortical neurons suggesting that it is not a pan neuronal enhancer. ETS factor-binding sites in the enhancer are required for its retinal neuron activity. These findings suggest that ETS factor interactions with beta 43' control retinal neuron expression of certain nAchR subtypes. PMID- 11734553 TI - A role for peroxisome proliferator-activated receptor alpha (PPARalpha ) in the control of cardiac malonyl-CoA levels: reduced fatty acid oxidation rates and increased glucose oxidation rates in the hearts of mice lacking PPARalpha are associated with higher concentrations of malonyl-CoA and reduced expression of malonyl-CoA decarboxylase. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a nuclear receptor transcription factor that has an important role in controlling cardiac metabolic gene expression. We determined whether mice lacking PPARalpha (PPARalpha (-/-) mice) have alterations in cardiac energy metabolism. Rates of palmitate oxidation were significantly decreased in isolated working hearts from PPARalpha (-/-) hearts compared with hearts from age-matched wild type mice (PPARalpha (+/+) mice), (62 +/- 12 versus 154 +/- 65 nmol/g dry weight/min, respectively, p < 0.05). This was compensated for by significant increases in the rates of glucose oxidation and glycolysis. The decreased fatty acid oxidation in PPARalpha (-/-) hearts was associated with increased levels of cardiac malonyl CoA compared with PPARalpha (+/+) hearts (15.15 +/- 1.63 versus 7.37 +/- 1.31 nmol/g, dry weight, respectively, p < 0.05). Since malonyl-CoA is an important regulator of cardiac fatty acid oxidation, we also determined if the enzymes that control malonyl-CoA levels in the heart are under transcriptional control of PPARalpha. Expression of both mRNA and protein as well as the activity of malonyl CoA decarboxylase, which degrades malonyl-CoA, were significantly decreased in the PPARalpha (-/-) hearts. In contrast, the expression and activity of acetyl CoA carboxylase, which synthesizes malonyl-CoA and 5'-AMP-activated protein kinase, which regulates acetyl-CoA carboxylase, were not altered. Glucose transporter expression (GLUT1 and GLUT4) was not different between PPARalpha (-/ ) and PPARalpha (+/+) hearts, suggesting that the increase in glycolysis and glucose oxidation in the PPARalpha null mice was not due to direct effects on glucose uptake but rather was occurring secondary to the decrease in fatty acid oxidation. This study demonstrates that PPARalpha is an important regulator of fatty acid oxidation in the heart and that this regulation of fatty acid oxidation may in part occur due to the transcriptional control of malonyl-CoA decarboxylase. PMID- 11734554 TI - The amino acid sequence of a plastid protein is developmentally regulated by RNA editing. AB - RNA editing in plant organelles post-transcriptionally alters single nucleotides by C-to-U or U-to-C conversions at highly specific sites. Plant editing is generally viewed as a repair mechanism acting at the transcript level by restoring conserved amino acid residues. Here we report that an editing reaction within the ndhB transcript (encoding a plastid NAD(P)H dehydrogenase subunit) is strictly dependent on active photosynthesis. Employing non-photosynthetic mutants, we show that in the absence of photosynthesis, the site remains unedited, whereas it is fully edited when the photosynthetic apparatus is intact. Moreover, the site also remains unedited during the etiolated stage of seedling development, suggesting that two different NdhB proteins are synthesized under photosynthetic versus non-photosynthetic conditions. This is the first case where RNA editing in plants appears to regulate gene expression qualitatively, resulting in the production of two different proteins from one and the same gene in a developmental stage-dependent manner. PMID- 11734555 TI - A high affinity HSF-1 binding site in the 5'-untranslated region of the murine tumor necrosis factor-alpha gene is a transcriptional repressor. AB - Tumor necrosis factor-alpha (TNFalpha) is a pivotal early mediator of host defenses that is essential for survival in infections. We previously reported that exposing macrophages to febrile range temperatures (FRT) (38.5-40 degrees C) markedly attenuates TNFalpha expression by causing abrupt and premature cessation of transcription. We showed that this inhibitory effect of FRT is mediated by an alternatively activated repressor form of heat shock factor 1 (HSF-1) and that a fragment of the TNFalpha gene comprising a minimal 85-nucleotide (nt) proximal promoter and the 138-nt 5'-untranslated region (UTR) was sufficient for mediating this effect. In the present study we have used an electrophoretic mobility shift assay (EMSA) to identify a high affinity binding site for HSF-1 in the 5'-UTR of the TNFalpha gene and have used a chromosome immunoprecipitation assay to show that HSF-1 binds to this region of the endogenous TNFalpha gene. Mutational inactivation of this site blocks the inhibitory effect of overexpressed HSF-1 on activity of the minimal TNFalpha promoter (-85/+138) in Raw 264.7 murine macrophages, identifying this site as an HSF-1-dependent repressor. However, the same mutation fails to block repression of a full-length (-1080/+138) TNFalpha promoter construct by HSF-1 overexpression, and HSF-1 binds to upstream sequences in the regions -1080/-845, -533/-196, and -326/-39 nt in EMSA, suggesting that additional HSF-1-dependent repressor elements are present upstream of the minimal -85-nt promoter. Furthermore, although mutation of the HSF-1 binding site in the minimal TNFalpha promoter construct abrogates HSF-1-mediated repression, the same mutation fails to abrogate repression of this construct by high levels of HSF-1 overexpression or exposure to 39.5 degrees C. This suggests that HSF-1 might repress TNFalpha transcription through redundant mechanisms, some of which might not require high affinity binding of HSF-1. PMID- 11734556 TI - The creatine kinase system is essential for optimal refill of the sarcoplasmic reticulum Ca2+ store in skeletal muscle. AB - Muscle function depends on an adequate ATP supply to sustain the energy consumption associated with Ca(2+) cycling and actomyosin sliding during contraction. In this regulation of energy homeostasis, the creatine kinase (CK) circuit for high energy phosphoryl transfer between ATP and phosphocreatine plays an important role. We earlier established a functional connection between the activity of the CK system and Ca(2+) homeostasis during depolarization and contractile activity of muscle. Here, we show how CK activity is coupled to the kinetics of spontaneous and electrically induced Ca(2+) transients in the sarcoplasm of myotubes. Using the UV ratiometric Ca(2+) probe Indo-1 and video rate confocal microscopy in CK-proficient and -deficient cultured cells, we found that spontaneous and electrically induced transients were dependent on ryanodine sensitive Ca(2+) release channels, sarcoplasmic/endoplasmic reticulum Ca(2+) ATPase pumps, extracellular calcium, and functional mitochondria in both cell types. However, at increasing sarcoplasmic Ca(2+) load (induced by electrical stimulation at 0.1, 1, and 10 Hz), the Ca(2+) removal rate and the amount of Ca(2+) released per transient were gradually reduced in CK-deficient (but not wild-type) myotubes. We conclude that the CK/phosphocreatine circuit is essential for efficient delivery of ATP to the sarcoplasmic/endoplasmic reticulum Ca(2+) ATPase pumps and thereby directly influences sarcoplasmic reticulum refilling and the kinetics of the sarcoplasmic Ca(2+) signals. PMID- 11734557 TI - SYT associates with human SNF/SWI complexes and the C-terminal region of its fusion partner SSX1 targets histones. AB - A global transcriptional co-activator, the SNF/SWI complex, has been characterized as a chromatin remodeling factor that enhances accessibility of the transcriptional machinery to DNA within a repressive chromatin structure. On the other hand, mutations in some human SNF/SWI complex components have been linked to tumor formation. We show here that SYT, a partner protein generating the synovial sarcoma fusion protein SYT-SSX, associates with native human SNF/SWI complexes. The SYT protein has a unique QPGY domain, which is also present in the largest subunits, p250 and the newly identified homolog p250R, of the corresponding SNF/SWI complexes. The C-terminal region (amino acids 310-387) of SSX1, comprising the SSX1 portion of the SYT-SSX1 fusion protein, binds strongly to core histones and oligonucleosomes in vitro and directs nuclear localization of a green fluorescence protein fusion protein. Experiments with serial C terminal deletion mutants of SSX1 indicate that these properties map to a common region and also correlate with the previously demonstrated anchorage-independent colony formation activity of SYT-SSX in Rat 3Y1 cells. These data suggest that SYT-SSX interferes with the function of either the SNF/SWI complexes or another SYT-interacting co-activator, p300, by changing their targeted localization or by directly inhibiting their chromatin remodeling activities. PMID- 11734558 TI - Identification of amino acid residues critical for LD78beta, a variant of human macrophage inflammatory protein-1alpha, binding to CCR5 and inhibition of R5 human immunodeficiency virus type 1 replication. AB - In an attempt to determine which amino acid(s) of LD78beta, a variant of human macrophage inflammatory protein-1alpha, plays a critical role in the interaction with CCR5, we generated six LD78beta variants with an amino acid substituted to Ala at the NH(2) terminus of LD78beta. There was no significant difference in eliciting Ca(2+) flux and chemotaxis among the variants with the exception of LD78beta(T9A) showing a substantially reduced activity. The comparative order for human immunodeficiency virus type 1 (HIV-1) replication inhibition was: LD78beta(P8A) > LD78beta(D6A) > LD78beta(WT), LD78beta(L3A) > LD78beta(T7A), LD78beta(P2A) > LD78beta(T9A). In binding inhibition assays of LD78beta variants using 2D7 monoclonal antibody and (125)I-labeled macrophage inflammatory protein 1alpha, the comparative order was: LD78beta(P8A), LD78beta(D6A) > LD78beta(WT) > LD78beta(L3A) > LD78beta(T7A) > LD78beta(T9A), LD78betaP2A). The order for CCR5 down-regulation induction was comparable to that for binding inhibition. The present data suggest that Pro-2, Asp-6, Pro-8, and Thr-9 are critical for LD78beta binding to CCR5 and HIV-1 replication inhibition, and that LD78beta binding to CCR5, regardless of affinity, is sufficient for the initial signal transduction of LD78beta, whereas the greater anti-HIV-1 activity requires the greater magnitude of binding. The data also suggest that LD78beta variants with appropriate amino acid substitution(s) such as LD78beta(D6A) and LD78beta(P8A) may represent effective chemokine-based anti-HIV-1 therapeutics while preserving LD78beta-CCR5 interactions. PMID- 11734559 TI - Identification of a novel homotypic interaction motif required for the phosphorylation of receptor-interacting protein (RIP) by RIP3. AB - Receptor-interacting protein (RIP), a Ser/Thr kinase component of the tumor necrosis factor (TNF) receptor-1 signaling complex, mediates activation of the nuclear factor kappaB (NF-kappaB) pathway. RIP2 and RIP3 are related kinases that share extensive sequence homology with the kinase domain of RIP. Unlike RIP, which has a C-terminal death domain, and RIP2, which has a C-terminal caspase activation and recruitment domain, RIP3 possesses a unique C terminus. RIP3 binds RIP through this unique C-terminal segment to inhibit RIP- and TNF receptor-1 mediated NF-kappaB activation. We have identified a unique homotypic interaction motif at the C terminus of both RIP and RIP3 that is required for their association. Sixty-four amino acids within RIP3 and 88 residues within RIP are sufficient for interaction of the two proteins. This interaction is a prerequisite for RIP3-mediated phosphorylation of RIP and subsequent attenuation of TNF-induced NF-kappaB activation. PMID- 11734560 TI - Efficiency and accuracy of SOS-induced DNA polymerases replicating benzo[a]pyrene 7,8-diol 9,10-epoxide A and G adducts. AB - Nucleotide incorporation fidelity, mismatch extension, and translesion DNA synthesis efficiencies were determined using SOS-induced Escherichia coli DNA polymerases (pol) II, IV, and V to copy 10R and 10S isomers of trans-opened benzo[a]pyrene-7,8-diol 9,10-epoxide (BaP DE) A and G adducts. A-BaP DE adducts were bypassed by pol V with moderate accuracy and considerably higher efficiency than by pol II or IV. Error-prone pol V copied G-BaP DE-adducted DNA poorly, forming A*G-BaP DE-S and -R mismatches over C*G-BaP DE-S and -R correct matches by factors of approximately 350- and 130-fold, respectively, even favoring G*G BaP DE mismatches over correct matches by factors of 2-4-fold. In contrast, pol IV bypassed G-BaP DE adducts with the highest efficiency and fidelity, making misincorporations with a frequency of 10(-2) to 10(-4) depending on sequence context. G-BaP DE-S-adducted M13 DNA yielded 4-fold fewer plaques when transfected into SOS-induced DeltadinB (pol IV-deficient) mutant cells compared with the isogenic wild-type E. coli strain, consistent with the in vitro data showing that pol IV was most effective by far at copying the G-BaP DE-S adduct. SOS polymerases are adept at copying a variety of lesions, but the relative contribution of each SOS polymerase to copying damaged DNA appears to be determined by the lesion's identity. PMID- 11734561 TI - S-adenosylmethionine decarboxylase from Leishmania donovani. Molecular, genetic, and biochemical characterization of null mutants and overproducers. AB - The polyamine biosynthetic enzyme, S-adenosylmethionine decarboxylase (ADOMETDC) has been advanced as a potential target for antiparasitic chemotherapy. To investigate the importance of this protein in a model parasite, the gene encoding ADOMETDC has been cloned and sequenced from Leishmania donovani. The Delta adometdc null mutants were created in the insect vector form of the parasite by double targeted gene replacement. The Delta adometdc strains were incapable of growth in medium without polyamines; however, auxotrophy could be rescued by spermidine but not by putrescine, spermine, or methylthioadenosine. Incubation of Delta adometdc parasites in medium lacking polyamines resulted in a drastic increase of putrescine and glutathione levels with a concomitant decrease in the amounts of spermidine and the spermidine-containing thiol trypanothione. Parasites transfected with an episomal ADOMETDC construct were created in both wild type and Delta adometdc parasites. ADOMETDC overexpression abrogated polyamine auxotrophy in the Delta adometdc L. donovani. In addition, ADOMETDC overproduction in wild type parasites alleviated the toxic effects of 5'-(((Z)-4 amino-2-butenyl)methylamino)-5'-deoxyadenosine (MDL 73811), but not pentamidine, berenil, or methylglyoxyl bis(guanylhydrazone), all inhibitors of ADOMETDC activities in vitro. The molecular, biochemical, and genetic characterization of ADOMETDC establishes that it is essential in L. donovani promastigotes and a potential target for therapeutic validation. PMID- 11734562 TI - The Yin Yang 1 transcription factor associates with ribonucleoprotein (mRNP) complexes in the cytoplasm of Xenopus oocytes. AB - Yin Yang 1 (YY1) is a multifunctional transcription factor that activates, represses, or initiates transcription of a diverse assortment of genes. Previous studies suggest a role for YY1 in cellular growth and differentiation, but its biological function during development of the vertebrate oocyte or embryo remains to be determined. We recently showed that YY1 is abundantly expressed throughout oogenesis and early embryonic stages of Xenopus, but it is sequestered in the cytoplasm and does not function directly in transcriptional regulation. In the present study we used a series of biochemical analyses to explore the potential function of YY1 in the oocyte cytoplasm. YY1 was isolated from oocyte lysates by oligo(dT)-cellulose chromatography, suggesting that it associates with maternally expressed mRNA in vivo. RNA mobility shift assays demonstrate that endogenous YY1 binds to labeled histone mRNA. Size exclusion chromatography of oocyte lysates revealed that YY1 exists in high molecular mass complexes in the range of 480 kDa. Destruction of endogenous RNA by RNase treatment of lysates, abolished the binding of YY1 to oligo(dT)-cellulose and resulted in redistribution from 480-kDa complexes to the monomeric form. Microinjection of RNase directly into the cytoplasm released YY1 from 480-kDa complexes and unmasked its DNA-binding activity, but did not promote translocation to the nucleus. These results provide evidence that YY1 is a component of ribonucleoprotein (mRNP) complexes in the Xenopus oocyte, indicating a novel function for YY1 in the storage or metabolism of maternal transcripts. PMID- 11734563 TI - Activation of p59(Fyn) leads to melanocyte dedifferentiation by influencing MKP-1 regulated mitogen-activated protein kinase signaling. AB - Malignant melanoma is a cancer whose incidence is rising rapidly, but the mechanism by which normal melanocytes become malignant in vivo is still little understood. In the course of melanoma progression, a fraction of cells often becomes depigmented, which reflects the loss of the balance between mitogenic activities and differentiation in those pigment cells. A key factor involved in differentiation in pigment cells is mitogen-activated protein kinase (MAPK). However, because both activation and inhibition of MAPK signaling is known to correlate with differentiation, its function in pigment cells is still unclear. We investigated the role of MAPK signaling in pigment cells using the melanoma inducing receptor tyrosine kinase Xmrk. Xmrk signaling in mouse melanocytes suppressed differentiation and induced a transformed phenotype. We found that this was based on sustained MAPK activation caused by low and transient expression of MAPK-phosphatase MKP-1. The Src kinase p59(Fyn) was thereby identified as being crucial for the receptor-mediated suppression of differentiation by down-regulating MKP-1 expression. Our findings reveal a novel mechanism of regulating the balance between differentiation and proliferation based on a Src kinase-modified MAPK activity. Moreover, they point to a new role for Src kinases in dedifferentiation and transformation of pigment cells. PMID- 11734564 TI - Glutathione dependence of caspase-8 activation at the death-inducing signaling complex. AB - Apoptosis triggered by the death receptor CD95 (APO-1 or Fas) is pivotal for the homeostasis of the immune system. We investigated differential effects of glutathione depletion on CD95-triggered apoptosis in T and B cell lines as well as the glutathione dependence of caspase-8 activation. In B lymphoblastoid SKW6.4 cells, CD95-mediated apoptosis was prevented upstream of caspase-8 activation and caspase-3-like activity after acute glutathione depletion by diethyl maleate or cis-chloro-dinitrobenzene. Immunoprecipitation of the death-inducing signaling complex (DISC) revealed that the DISC was still formed in the glutathione depleted state. The first cleavage step of procaspase-8 activation at the DISC, however, was inhibited. Accordingly, under cell-free conditions, radiolabeled procaspase-8 was processed at the immunoprecipitated DISC only after the addition of exogenous dithiothreitol or reduced glutathione. We also observed suppression of CD95-mediated apoptosis in glutathione-depleted CEM and H9 cells. Notably, Jurkat cells still died upon CD95 engagement under this condition, displaying incomplete nuclear fragmentation and a partial switch to necrosis; this may be explained by reduced cytochrome c/dATP-mediated caspase activation observed in cytosol from glutathione-depleted Jurkat cytosol. Our data indicate that the activation of caspase-8 at the DISC and hence CD95-mediated apoptosis induction shows a cell-specific requirement for intracellular glutathione. PMID- 11734565 TI - Catalysis of ACAT may be completed within the plane of the membrane: a working hypothesis. AB - Two ACAT sharing protein sequence homology near their C termini have been identified. Both proteins may span the endoplasmic reticulum (ER) membrane several times. There is good evidence implicating the role of ACAT1 in macrophage foam cell formation, and ACAT2 in intestinal cholesterol absorption. On the other hand, the functional roles of ACAT1 and ACAT2 in the VLDL or chylomicron assembly process are less clear. It is possible that both enzymes are able to form lipid droplets (which are present in the cytoplasm), and participate in lipoprotein assembly (which occurs in the ER lumen). To link the site of ACAT catalysis with its function, we propose that part of the ACAT catalytic site may reside within the lipid bilayer, allowing catalysis to be completed within the plane of the membrane. Cholesteryl esters (CE) produced in situ may burst into cytoplasmic lipid droplets, carrying phospholipid monolayers as their outer coats. In cells engaged in lipoprotein assembly and secretion, CE in the bilayer may be recognized by the specific protein microsomal triacylglycerol transfer protein (MTP), reaching out from the lumenal side of the membrane. MTP then lipidates the growing apolipoprotein B (apoB) chain with CE and TG during the early stages of apoB lipoprotein assembly. PMID- 11734566 TI - Apoptosis-induced release of mature sterol regulatory element-binding proteins activates sterol-responsive genes. AB - It is well established that during the execution of the apoptotic cascade, activated caspase 3 releases sterol regulatory element-binding proteins (SREBP) from the membrane of the endoplasmic reticulum in a proteolytic reaction that is distinct from their normal sterol-dependent activation. However, it is not known whether these transcription factors are capable of activating sterol-responsive genes under such conditions. The construction of SRE expression vectors has permitted characterization of the apoptotic activation of SREBP. Cell lines stably expressing the plasma membrane marker CD32, or GFP, under the control of the SRE promoter were shown to modulate SRE gene expression on the basis of the levels of available sterols. However, during the induction of apoptosis, expression of CD32 and GFP was highly induced, even in the presence of ample sterols. Apoptotic induction of sterol-regulated genes was due to activation of caspase 3 and was impervious to treatment with sphingomyelinase, indicating that activation of SRE genes during apoptosis is sterol independent. Further characterization of this apoptotic response indicated that sterol-regulated genes are activated at an early stage in the apoptotic cascade, preceding the externalization of phosphatidylserine on the plasma membrane of apoptotic cells. These results suggest that activation of sterol-responsive genes early during apoptosis may play a role in the proper execution of this program. PMID- 11734567 TI - Distribution, transport, and degradation of apolipoprotein B-100 in HepG2 cells. AB - The transport of apolipoprotein B (apoB) between the endoplasmic reticulum (ER) and Golgi was studied in puromycin-synchronized HepG2 cells, using an antibody that could distinguish between apoB in ER and Golgi compartments. In cells with normal ER-to-Golgi transport, both albumin and apoB colocalized throughout the ER and appeared as intense, compact signals in Golgi. When ER-to-Golgi transport was blocked with brefeldin A, apoB and albumin remained colocalized in the ER network and three-dimensional constructed images showed more intense signals for both proteins in a central, perinuclear region of the ER. When protein synthesis was stopped in cells with brefeldin A-inhibited ER-to-Golgi transport, apoB degradation was visualized as a homogeneous decrease in fluorescence signal intensity throughout the ER that could be slowed with clasto-lactacystin beta lactone, a proteasome inhibitor. Incubation of cells with CP-10447, an inhibitor of microsomal triglyceride transfer protein, inhibited apoB, but not albumin, transport from ER to Golgi. Nanogold immunoelectron microscopy of digitonin permeabilized cells showed proteasomes in close proximity to the cytosolic side of the ER membrane. Thus, newly synthesized apoB is localized throughout the entire ER and degraded homogeneously, most likely by neighboring proteasomes located on the cytosolic side of the ER membrane. Although albumin is colocalized with apoB in the ER, as expected, it was not targeted for ER-associated proteasomal degradation. PMID- 11734568 TI - Molecular species composition of rat liver phospholipids by ESI-MS/MS: the effect of chromatography. AB - Using electrospray ionization tandem mass spectrometry (ESI-MS/MS) this study shows that the loss of glycerophospholipid (GPL) after chromatography was unevenly distributed across the GPL molecular species. Both TLC and HPLC caused a preferential loss of GPL with 0 to 3 double bonds: 20% and 7.2% for choline glycerophosphates (PC) and 19.7% and 7.5% for ethanolamine glycerophosphates (PE), respectively. A consequence of these losses was that GPLs containing fatty acids with four or more double bonds had a greater contribution to the total after chromatography. ESI-MS/MS analysis also showed that PC molecular species with four or more double bonds migrated at the front of the TLC band of PCs. GPLs extracted from TLC plates occasionally contained PCs that were smaller than those in the original extract. These low molecular mass PCs were easily reduced to alcohols and formed derivatives with 2,4-dinitrophenylhydrazine, suggesting that aldehydes were generated by the oxidation of unsaturated fatty acids. Directly analyzing lipid extracts by ESI-MS/MS without preliminary chromatographic separation gives an accurate distribution of GPL molecular species in lipid mixtures. However, the ionization of the phospholipids in the electrospray jet maximized at relatively low concentrations of GPL. There was a linear response between phospholipid mass and ion intensity for concentrations around 1-2 nmol/ml for both PC and PE. The total ion intensity continued to increase with concentrations above 1-2 nmol/ml, but the response was non-linear. PMID- 11734569 TI - Scavenger receptor class B type I affects cholesterol homeostasis by magnifying cholesterol flux between cells and HDL. AB - Results from several laboratories clearly indicate that expression of scavenger receptor class B type I (SR-BI) enhances the bidirectional flux of cholesterol between cells and lipoproteins. Because the activity of HMG-CoA reductase, the key enzyme in cholesterol biosynthesis, is regulated by cell cholesterol content, we designed experiments to investigate the effect of SR-BI expression on the activity of this enzyme and on net cellular cholesterol mass. In addition, we compared the function of SR-BI with its human homolog, CD36 and LIMPII analogous 1. Our experiments demonstrate that both receptors enhance the flux of unesterified or free cholesterol bidirectionally, down a concentration gradient. Receptor-mediated cholesterol flux can effectively modulate multiple aspects of cellular cholesterol metabolism, including the pool that regulates the activity of HMG-CoA reductase. We also found that constitutive expression of SR-BI alters the steady state level of cellular cholesterol and phospholipid when SR-BI expressing cells are maintained in medium containing serum lipoproteins. All of these effects are proportional to the level of receptor on the cell surface. These data indicate that the level of SR-BI expression determines both the rate of free cholesterol flux and the steady state level of cellular cholesterol. PMID- 11734570 TI - A novel pathway for lipid biosynthesis: the direct acylation of glycerol. AB - The acylation of glycerol-3-phosphate by acyl-CoA is regarded as the first committed step for the synthesis of the lipoidal moiety in glycerolipids. The direct acylation of glycerol in mammalian tissues has not been demonstrated. In this study, lipid biosynthesis in myoblasts and hepatocytes was reassessed by conducting pulse-chase experiments with [1,3-(3)H]glycerol. The results demonstrated that a portion of labeled glycerol was directly acylated to form monoacylglycerol and, subsequently, diacylglycerol and triacylglycerol. The direct acylation of glycerol became more prominent when the glycerol-3-phosphate pathway was attenuated or when exogenous glycerol levels became elevated. Glycerol:acyl-CoA acyltransferase activity, which is responsible for the direct acylation of glycerol, was detected in the microsomal fraction of heart, liver, kidney, skeletal muscle, and brain tissues. The enzyme from pig heart microsomes displayed optimal activity at pH 6.0 and the preference for arachidonyl-CoA as the acyl donor. The apparent K(m) values for glycerol and arachidonyl-CoA were 1.1 mM and 0.17 mM, respectively. The present study demonstrates the existence of a novel lipid biosynthetic pathway that may be important during hyperglycerolemia produced in diabetes or other pathological conditions. PMID- 11734571 TI - Identification of the peroxisomal beta-oxidation enzymes involved in the biosynthesis of docosahexaenoic acid. AB - DHA (C22:6n-3) is an important PUFA implicated in a number of (patho)physiological processes. For a long time, the exact mechanism of DHA formation has remained unclear, but now it is known that it involves the production of tetracosahexaenoic acid (C24:6n-3) from dietary linolenic acid (C18:3n-3) via a series of elongation and desaturation reactions, followed by beta-oxidation of C24:6n-3 to C22:6n-3. Although DHA is deficient in patients lacking peroxisomes, the intracellular site of retroconversion of C24:6n-3 has remained controversial. By making use of fibroblasts from patients with defined mitochondrial and peroxisomal fatty acid oxidation defects, we show in this article that peroxisomes, and not mitochondria, are involved in DHA formation by catalyzing the beta-oxidation of C24:6n-3 to C22:6n-3. Additional studies of fibroblasts from patients with X-linked adrenoleukodystrophy, straight-chain acyl CoA oxidase (SCOX) deficiency, d-bifunctional protein (DBP) deficiency, and rhizomelic chondrodysplasia punctata type 1, and of fibroblasts from l bifunctional protein and sterol carrier protein X (SCPx) knockout mice, show that the main enzymes involved in beta-oxidation of C24:6n-3 to C22:6n-3 are SCOX, DBP, and both 3-ketoacyl-CoA thiolase and SCPx. These findings are of importance for the treatment of patients with a defect in peroxisomal beta-oxidation. PMID- 11734572 TI - Plasma lipids in Turkish children: impact of puberty, socioeconomic status, and nutrition on plasma cholesterol and HDL. AB - In Turkish adults, HDL cholesterol (HDL-C) levels are 10-15 mg/dl lower than those of adults in western Europe and the United States. In this study, we determined whether HDL-C levels in Turks are low from birth to adulthood and assessed the effect of socioeconomic status (SES) on plasma lipids and lipoproteins. Analyses of cord blood from 105 Turkish newborns showed low levels of plasma cholesterol ( approximately 60 mg/dl) and HDL-C (approximately 30 mg/dl), consistent with results from other Western ethnic groups. Prepubescent 8- to 10-year-old Turkish boys and girls of upper (n = 82) and lower (n = 143) SES had high HDL-C levels (50-60 mg/dl) similar to those of western European children. However, the cholesterol (154-158 mg/dl) and HDL-C (55-58 mg/dl) levels of upper SES children were approximately 25 and approximately 12 mg/dl higher, respectively, than those of lower SES children. Height, weight, skinfold thickness, and estimated body fat were greater in the upper SES children and appeared to reflect dietary differences. Upper SES children consumed more total fat (approximately 35% vs. 25% of total calories), including more saturated fat of animal origin, and less carbohydrate (approximately 50% vs. 62% of total calories), consistent with their elevated plasma cholesterol levels. Carbohydrate intake correlated inversely with the HDL-C level. The HDL-C levels in the prepubescent children, especially those of higher SES, who consumed diets more like western Europeans, decreased markedly to adult levels, with males exhibiting a approximately 20 mg/dl decrease (from 58 to 37 mg/dl) and females a approximately 13 mg/dl decrease (from 55 to 42 mg/dl). SES did not affect HDL-C levels in adults. The profound decrease may reflect alterations in androgen/estrogen balance in Turks at puberty and a modulation of hepatic lipase affecting HDL-C levels. PMID- 11734573 TI - Reduced HDL particle size as an additional feature of the atherogenic dyslipidemia of abdominal obesity. AB - Reduced plasma HDL cholesterol concentration has been associated with an increased risk of coronary heart disease. However, a low HDL cholesterol concentration is usually not observed as an isolated disorder because this condition is often accompanied by additional metabolic alterations. The objective of this study was to document the relevance of assessing HDL particle size as another feature of the atherogenic dyslipidemia found among subjects with visceral obesity and insulin resistance. For that purpose, an average HDL particle size was computed by calculating an integrated HDL particle size using nondenaturing 4-30% gradient gel electrophoresis. Potential associations between this average HDL particle size versus morphometric and metabolic features of visceral obesity were examined in a sample of 238 men. Results of this study indicated that HDL particle size was a significant correlate of several features of an atherogenic dyslipidemic profile such as increased plasma TG, decreased HDL cholesterol, high apolipoprotein B, elevated cholesterol/HDL cholesterol ratio, and small LDL particles as well as increased levels of visceral adipose tissue (AT) (0.33 < or = absolute value of r < or = 0.61, P < 0.0001). Thus, men with large HDL particles had a more favorable plasma lipoprotein-lipid profile compared with those with smaller HDL particles. Furthermore, men with large HDL particles were also characterized by reduced overall adiposity and lower levels of visceral AT as well as reduced insulinemic-glycemic responses to an oral glucose load. In conclusion, small HDL particle size appears to represent another feature of the high TG- low HDL cholesterol dyslipidemia found in viscerally obese subjects characterized by hyperinsulinemia. PMID- 11734574 TI - Eicosapentaenoic acid and docosahexaenoic acid modulate MAP kinase (ERK1/ERK2) signaling in human T cells. AB - This study was conducted on human Jurkat T cell lines to elucidate the role of EPA and DHA, n-3 PUFA, in the modulation of two mitogen-activated protein (MAP) kinases, that is, extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2). The n-3 PUFA alone failed to induce phosphorylation of ERK1/ERK2. We stimulated the MAP kinase pathway with anti-CD3 antibodies and phorbol 12-myristate 13 acetate (PMA), which act upstream of the MAP kinase (MAPK)/ERK kinase (MEK) as U0126, an MEK inhibitor, abolished the actions of these two agents on MAP kinase activation. EPA and DHA diminished the PMA- and anti-CD3-induced phosphorylation of ERK1/ERK2 in Jurkat T cells. In the present study, PMA acts mainly via protein kinase C (PKC) whereas anti-CD3 antibodies act via PKC-dependent and -independent mechanisms. Furthermore, DHA and EPA inhibited PMA-stimulated PKC enzyme activity. EPA and DHA also significantly curtailed PMA- and ionomycin-stimulated T cell blastogenesis. Together these results suggest that EPA and DHA modulate ERK1/ERK2 activation upstream of MEK via PKC-dependent and -independent pathways and that these actions may be implicated in n-3 PUFA-induced immunosuppression. PMID- 11734575 TI - Increased plasma apoA-IV level is a marker of abnormal postprandial lipemia: a study in normoponderal and obese subjects. AB - Plasma apolipoprotein A-IV (apoA-IV) levels are found elevated in hypertriglyceridemic patients. However, the relationship between plasma apoA-IV level and postprandial lipemia is not well known and remains to be elucidated. Thus, our objective was to study the relationship between plasma apoA-IV and postprandial TG after an oral fat load test (OFLT). Plasma apoA-IV was measured at fast and during an OFLT in 16 normotriglyceridemic, normoglucose-tolerant android obese subjects (BMI = 34.6 +/- 2.9 kg/m(2)) and 30 normal weight controls (BMI = 22.2 +/- 2.3 kg/m(2)). In spite of not statistically different fasting plasma TG levels in controls and obese patients, the former group showed an altered TG response after OFLT, featuring increased nonchylomicron TG area under the curve (AUC) compared with controls (516 +/- 138 vs. 426 +/- 119 mmol/l x min, P < 0.05). As compared to controls, obese patients showed increased apoA-IV levels both at fast (138.5 +/- 22.4 vs. 124.0 +/- 22.8 mg/l, P < 0.05) and during the OFLT (apoA-IV AUC: 79,833 +/- 14,281 vs. 68,176 +/- 17,463 mg/l x min, P < 0.05). Among the whole population studied, as among the control and obese subgroups, fasting plasma apoA-IV correlated significantly with AUC of plasma TG (r = 0.60, P < 0.001), AUC of chymomicron TG (r = 0.45, P < 0.01), and AUC of nonchylomicron TG (r = 0.62, P < 0.001). In the multivariate analysis, fasting apoA-IV level constituted an independent and highly significant determinant of AUC of plasma TG, AUC of chymomicron TG, AUC of nonchylomicron TG, and incremental AUC of plasma TG. In conclusion, we show a strong link between fasting apoA-IV and postprandial TG metabolism. Plasma fasting apoA-IV is shown to be a good marker of TG response after an OFLT, providing additional information on post-load TG response in conjunction with other known factors such as fasting TGs. PMID- 11734576 TI - Oxidized plant sterols in human serum and lipid infusions as measured by combined gas-liquid chromatography-mass spectrometry. AB - Some oxidized forms of cholesterol (oxysterols) are thought to be atherogenic and cytotoxic. Because plant sterols are structurally related to cholesterol, we examined whether oxidized plant sterols (oxyphytosterols) could be identified in human serum and soy-based lipid emulsions. We first prepared both deuterated and nondeuterated reference compounds. We then analyzed by gas-liquid chromatography mass spectrometry the oxyphytosterol concentrations in serum from patients with phytosterolemia or cerebrotendinous xanthomatosis, in a pool serum and in two lipid emulsions. 7-Ketositosterol, 7 beta-hydroxysitosterol, 5 alpha, 6 alpha epoxysitosterol, 3 beta,5 alpha,6 beta-sitostanetriol, and probably also 7 alpha hydroxysitosterol were present in markedly elevated concentrations in serum from phytosterolemic patients only. Also, campesterol oxidation products such as 7 alpha-hydroxycampesterol and 7 beta-hydroxycampesterol were found. Interestingly, sitosterol was oxidized for approximately 1.4% in phytosterolemic serum, which is rather high compared with the approximate 0.01% oxidatively modified cholesterol normally seen in human serum. The same oxyphytosterols were also found in two lipid emulsions in which the ratio of oxidized sitosterol to sitosterol varied between 0.038 and 0.041. In conclusion, we have shown that oxidized forms of plant sterols are present in serum from phytosterolemic patients and two frequently used soy-based lipid emulsions. Currently, it is unknown whether oxyphytosterols affect health, as has been suggested for oxysterols. However, 7 beta-hydroxycholesterol may be one of the more harmful oxysterols, and both sitosterol and campesterol were oxidized into 7 beta-hydroxysitosterol and 7 beta hydroxycampesterol. The relevance of these findings therefore deserves further exploration. PMID- 11734577 TI - Increased hepatic VLDL secretion, lipogenesis, and SREBP-1 expression in the corpulent JCR:LA-cp rat. AB - The corpulent JCR:LA-cp rat (cp/cp) is a useful model for study of the metabolic consequences of obesity and hyperinsulinemia. To assess the effect of hyperinsulinemia on VLDL secretion in this model, we measured rates of secretion of VLDL in perfused livers derived from cp/cp rats and their lean littermates. Livers of cp/cp rats secreted significantly greater amounts of VLDL triglyceride and apolipoprotein, compared with lean littermates. The content of apoB, apoE, and apoCs in both perfusate and plasma VLDL was greater in the cp/cp rat, as was the apolipoprotein (apo)C, apoA-I, and apoA-IV content of plasma HDL. Triglyceride content was also greater in cp/cp livers, as was hepatic lipogenesis and expression of lipogenic enzymes and sterol regulatory element binding protein 1 (SREBP-1). Hepatic mRNAs for apoE, and apoA-I were higher in livers of cp/cp rats. In contrast, the steady state levels of apoC-II, apoC-III, and apoB mRNAs were unchanged. Thus, livers of obese hyperinsulinemic cp/cp JCR:LA-cp rats secrete a greater number of VLDL particles that are enriched in triglyceride, apoE, and apoC. Greater secretion of VLDL in the cp/cp rat in part results from higher endogenous fatty acid synthesis, which in turn may occur in response to increased expression of the lipogenic enzyme regulator SREBP-1c. PMID- 11734578 TI - Fatty acid specificity of hormone-sensitive lipase. Implication in the selective hydrolysis of triacylglycerols. AB - The selective mobilization of fatty acids from white fat cells depends on their molecular structure, in particular the degree of unsaturation. The present study was designed to examine if the release of fatty acids by hormone-sensitive lipase (HSL) in vitro i) is influenced by the amount of unsaturation, ii) depends on the temperature, and iii) could explain the selective pattern of fatty acid mobilization and notably the preferential mobilization of certain highly unsaturated fatty acids. Recombinant rat and human HSL were incubated with a lipid emulsion. The hydrolysis of 35 individual fatty acids, ranging in chain length from 12 to 24 carbon atoms and in unsaturation from 0 to 6 double bonds was measured. Fatty acid composition of in vitro released NEFA was compared with that of fat cell triacylglycerols (TAG), the ratio % NEFA/% TAG being defined as the relative hydrolysis. The relative hydrolysis of individual fatty acids differed widely, ranging from 0.44 (24:1n-9) to 1.49 (18:1n-7) with rat HSL, and from 0.38 (24:1n-9) to 1.67 (18:1n-7) with human HSL. No major difference was observed between rat and human HSL. The relative release was dependent on the number of double bonds according to chain length. The amount of fatty acid released by recombinant rat HSL was decreased but remained robust at 4 degrees C compared with 37 degrees C, and the relative hydrolysis of some individual fatty acids was affected. The relative hydrolysis of fatty acids moderately, weakly, and highly mobilized by adipose tissue in vivo was similar and close to unity in vitro. We conclude that i) the release of fatty acids by HSL is only slightly affected by their degree of unsaturation, ii) the ability of HSL to efficiently and selectively release fatty acids at low temperature could reflect a cold adaptability for poikilotherms or hibernators when endogenous lipids are needed, and iii) the selectivity of fatty acid hydrolysis by HSL does not fully account for the selective pattern of fatty acid mobilization, but could contribute to explain the preferential mobilization of some highly unsaturated fatty acids compared with others. PMID- 11734579 TI - Isolation, quantitation, and characterization of a stable complex formed by Lp[a] binding to triglyceride-rich lipoproteins. AB - Lipoprotein [a] (Lp[a]) is a cholesterol-rich lipoprotein resembling LDL to which a large polymorphic glycoprotein, apolipoprotein [a] (apo[a]), is covalently coupled. Lp[a] usually exists as a free-standing particle in normolipidemic subjects; however, it can associate noncovalently with triglyceride-rich lipoproteins in hypertriglyceridemic (HTG) subjects. In this study, 10-78% of the Lp[a] present in five HTG subjects was found in the triglyceride-rich lipoprotein (TRL) fraction. The Lp[a]-TRL complex was resistant to dissociation by ultracentrifugation (UCF) alone, but was quantitatively dissociated by UCF in the presence of 100 mM proline. Of this dissociated Lp[a], 70-88% was in the form of a lipoprotein resembling conventional Lp[a]. Incubation of Lp[a]-depleted TRL with native Lp[a] resulted in a reconstituted Lp[a]-TRL complex that closely resembled the native isolates in all examined properties. Complex formation was inhibited by several compounds in the order proline > tranexamate > epsilon aminocaproate >> arginine > lysine. Neither plasminogen nor LDL inhibited binding of Lp[a] to TRL. We observed the preferential binding of Lp[a] containing higher apparent molecular weight apo[a] polymorphs to TRL both in native and reconstituted Lp[a]-TRL complexes. A disproportionate amount of Lp[a] was bound to the larger TRL particles. Although most apo[a] bound to TRL was in the form of conventional Lp[a] particles, lipid-free recombinant apo[a] was observed to bind TRL. These results provide unequivocal evidence of the existence of an Lp[a]-TRL complex under pathophysiologic conditions. The metabolic fate of the Lp[a]-TRL complex, which is more abundant in hypertriglyceridemia, may be different from that of conventional Lp[a], and may contribute uniquely to the progression or severity of cardiovascular disease. PMID- 11734580 TI - Walnut-enriched diet increases the association of LDL from hypercholesterolemic men with human HepG2 cells. AB - In a randomized, cross-over feeding trial involving 10 men with polygenic hypercholesterolemia, a control, Mediterranean-type cholesterol-lowering diet, and a diet of similar composition in which walnuts replaced approximately 35% of energy from unsaturated fat, were given for 6 weeks each. Compared with the control diet, the walnut diet reduced serum total and LDL cholesterol by 4.2% (P = 0.176), and 6.0% (P = 0.087), respectively. No changes were observed in HDL cholesterol, triglycerides, and apolipoprotein A-I levels or in the relative proportion of protein, triglycerides, phospholipids, and cholesteryl esters in LDL particles. The apolipoprotein B level declined in parallel with LDL cholesterol (6.0% reduction). Whole LDL, particularly the triglyceride fraction, was enriched in polyunsaturated fatty acids from walnuts (linoleic and alpha linolenic acids). In comparison with LDL obtained during the control diet, LDL obtained during the walnut diet showed a 50% increase in association rates to the LDL receptor in human hepatoma HepG2 cells. LDL uptake by HepG2 cells was correlated with alpha-linolenic acid content of the triglyceride plus cholesteryl ester fractions of LDL particles (r(2) = 0.42, P < 0.05). Changes in the quantity and quality of LDL lipid fatty acids after a walnut-enriched diet facilitate receptor-mediated LDL clearance and may contribute to the cholesterol-lowering effect of walnut consumption. PMID- 11734581 TI - Alcohol consumption stimulates early steps in reverse cholesterol transport. AB - Alcohol consumption is associated with increased HDL cholesterol levels, which may indicate stimulated reverse cholesterol transport. The mechanism is, however, not known. The aim of this study was to evaluate the effects of alcohol consumption on the first two steps of the reverse cholesterol pathway: cellular cholesterol efflux and plasma cholesterol esterification. Eleven healthy middle aged men consumed four glasses (40 g of alcohol) of red wine, beer, spirits (Dutch gin), or carbonated mineral water (control) daily with evening dinner, for 3 weeks, according to a 4 x 4 Latin square design. After 3 weeks of alcohol consumption the plasma ex vivo cholesterol efflux capacity, measured with Fu5AH cells, was raised by 6.2% (P < 0.0001) and did not differ between the alcoholic beverages. Plasma cholesterol esterification was increased by 10.8% after alcohol (P = 0.008). Changes were statistically significant after beer and spirits, but not after red wine consumption (P = 0.16). HDL lipids changed after alcohol consumption; HDL total cholesterol, HDL cholesteryl ester, HDL free cholesterol, HDL phospholipids and plasma apolipoprotein A-I all increased (P < 0.01). In conclusion, alcohol consumption stimulates cellular cholesterol efflux and its esterification in plasma. These effects were mostly independent of the kind of alcoholic beverage PMID- 11734582 TI - Preparation and incorporation of probe-labeled apoA-I for fluorescence resonance energy transfer studies of rHDL. AB - Apolipoprotein A-I (apoA-I), the major constituent of HDL, plays an essential role in regulating cholesterol metabolism, acting as the physiological activator of lecithin: cholesterol acyltransferase, which converts cholesterol to cholesterol ester. Thiol-reactive fluorescent probes attached to cysteine containing apoA-I mutants are currently being used to investigate the "LCAT active" conformation of lipid-bound apoA-I. Herein, we report new methodologies allowing rapid expression, fluorescent labeling, and recombinant HDL (rHDL) preparation for use in apoA-I in fluorescence resonance energy transfer (FRET) studies. Cysteine-containing mutant forms of human apoA-I were cloned into the pTYB12 vector containing a T7 promoter, a modified self-splicing protein element (intein), and a small affinity tag [chitin binding domain (CBD)]. The fusion proteins were expressed in Escherichia coli, isolated from cell lysates, and bound to a chitin-affinity column. Release of mature human apoA-I was initiated by the addition of DTT, which induced self-cleavage at the COOH terminus of the intein - CBD fusion protein. ApoA-I was further purified by Q-sepharose and then used for fluorescent probe labeling. Discoidal rHDL were then prepared with donor and/or acceptor labeled apoA-I and characterized with respect to their size, composition and ability to activate LCAT. PMID- 11734583 TI - Quantification of galactosylsphingosine in the twitcher mouse using electrospray ionization-tandem mass spectrometry. AB - Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive inherited neurodegenerative disorder caused by the deficiency of the lysosomal enzyme beta galactosylceramidase. The pathogenesis of the disorder has been proposed to arise from the accumulation of the cytotoxic metabolite galactosylsphingosine (psychosine). The twitcher mouse is a naturally occurring murine model of globoid cell leukodystrophy. We have developed a rapid, sensitive, and specific mass spectrometric method for determining the galactosylsphingosine concentration in the tissues of twitcher mice. Galactosylsphingosine is extracted from the tissues in methanol, isolated using strong cation-exchange and C18 solid-phase extraction chromatography, and then directly analyzed using electrospray ionization-tandem mass spectrometry. A lactosylsphingosine internal standard has been employed for quantification. The assay demonstrated significant accumulation of galactosylsphingosine in the brain, spinal cord, and kidney of twitcher mice. It is anticipated that this method may be of use in the monitoring of experimental therapies for globoid cell leukodystrophy. PMID- 11734584 TI - Breast cancer risk prediction: should nipple aspiration fluid cytology be incorporated into clinical practice? PMID- 11734585 TI - Hormone replacement therapy and adenoma recurrence: implications for its role in colorectal cancer risk. PMID- 11734586 TI - Beyond the Nobel Prize: cell cycle research offers new view of cancer. PMID- 11734588 TI - Clinical trial design changing as molecular targets gain popularity. PMID- 11734591 TI - U.S. tops list of most published clinical cancer research. PMID- 11734592 TI - Imaging facilities focus on small animal research. PMID- 11734593 TI - Regression of human T-cell leukemia virus type I (HTLV-I)-associated lymphomas in a rat model: peptide-induced T-cell immunity. AB - BACKGROUND: Human T-cell leukemia virus type I (HTLV-I) is etiologically linked to adult T-cell leukemia (ATL). The disease has a high mortality rate and is resistant to chemotherapy; therefore, immunologic approaches to treatment could be of interest. We have previously shown that athymic rats inoculated with a syngeneic (i.e., with the same genetic background) HTLV-I-infected T-cell line (FPM1-V1AX) develop ATL-like disease and that the transfer of T cells from normal syngeneic rats immunized with FPM1-V1AX cells prevents disease development. In this study, we further characterized the host antitumor immunity to explore the possibility of peptide-based vaccination against the ATL-like disease. METHODS: Immune T cells from rats immunized with FPM1-V1AX cells were analyzed for their phenotypes and cytotoxic properties. The epitope recognized by the T cells was analyzed by fine mapping. To evaluate the antitumor effects of a peptide-based vaccine, normal rats were immunized with synthetic oligopeptides corresponding to the epitope, the T cells were transferred to athymic rats inoculated with HTLV-I infected cells, and tumor size was monitored. RESULTS: Both CD4+ and CD8+ T-cell populations from rats immunized with FPM1-V1AX cells inhibited the growth of FPM1 V1AX cell-induced lymphomas in vivo. Long-term culture of splenic T cells from the immunized rats repeatedly resulted in establishment of CD8+ HTLV-I-specific cytotoxic T lymphocyte (CTL) lines restricted to the rat major histocompatibility complex class I molecule, RT1.A(l). The cytotoxicity of these lines was directed against the HTLV-I regulatory protein Tax and, specifically, against the epitope, amino acids 180-188 (GAFLTNVPY). Adoptive transfer of the Tax 180-188-specific CTL line or freshly prepared T cells from rats vaccinated with the Tax 180-188 oligopeptide prevented the development of FPM1-V1AX-cell induced lymphomas in athymic rats in comparison with control groups (two rats in each group). CONCLUSIONS: This study indicated a potential therapeutic effect of peptide-based vaccination against HTLV-I-induced lymphoproliferative disease. PMID- 11734594 TI - Role of fibroblast growth factor receptor signaling in prostate cancer cell survival. AB - BACKGROUND: Expression of fibroblast growth factors (FGFs) is increased in a substantial fraction of human prostate cancers in vivo and in prostate cancer cell lines. Altered FGF signaling can potentially have a variety of effects, including stimulating cell proliferation and inhibiting cell death. To determine the biologic significance of altered FGF signaling in human prostate cancer, we disrupted signaling by expression of a dominant-negative (DN) FGF receptor in prostate cancer cell lines. METHODS: PC-3, LNCaP, and DU145 prostate cancer cells were stably transfected with DN FGFR constructs, and LNCaP and DU145 cells were infected with a recombinant adenovirus expressing DN FGFR-1. The effect of DN FGFR-1 expression was assessed by colony-formation assays, cell proliferation assays, flow cytometry, and cytogenetic analysis. Key regulators involved in the G(2)-to-M cell cycle transition were assessed by western blotting to examine cyclin B1 expression and by in vitro kinase assay to assess cdc2 kinase activity. RESULTS: Stable transfection of the DN FGFR-1 construct inhibited colony formation by more than 99% in all three cell lines. Infection of LNCaP and DU145 prostate cancer cells with adenovirus expressing DN FGFR-1 led to extensive cell death within 48 hours. Flow cytometry and cytogenetic analysis revealed that the DN FGFR-1 receptor led to arrest in the G(2) phase of the cell cycle before cell death. Cyclin B1 accumulated in DN FGFR-1-infected LNCaP cells, but cdc2 kinase activity was decreased. CONCLUSIONS: These findings reveal an unexpected dependence of prostate cancer cells on FGF receptor signal transduction to traverse the G(2)/M checkpoint. The mechanism for the G(2) arrest is not clear. Our results raise the possibility that FGF-signaling antagonists might enhance the cell death induced by other prostate cancer therapies. PMID- 11734595 TI - Breast cancer risk in women with abnormal cytology in nipple aspirates of breast fluid. AB - BACKGROUND: We previously showed that women with abnormal cytology in breast fluid obtained by nipple aspiration had an increased relative risk (RR) of breast cancer compared with women from whom fluid was not obtained and with women whose fluid had normal cytology. This study extends the follow-up in the original study group (n = 4046) and presents the first follow-up for a second group of women (n = 3627). METHODS: We collected nipple aspirate fluid from women in the San Francisco Bay Area during the period from 1972 through 1991, classified the women according to the most severe epithelial cytology observed in fluid specimens, and determined breast cancer incidence through March 1999. We estimated RRs for breast cancer using Cox regressions, adjusting for age and year of study entry. All statistical tests were two-sided. RESULTS: For women in the first and second study groups, the median years of follow-up were 21 years and 9 years, respectively, and breast cancer incidences were 7.8% (285 cases in the 3633 women for whom breast cancer status could be determined) and 3.5% (115 of 3271), respectively. Compared with women from whom no fluid was obtained, whose incidences of breast cancer were 4.7% (39 of 825) and 3.3% (65 of 1950) for those in group 1 and group 2, respectively, incidences and adjusted RRs were 8.1% (34 of 422), with RR = 1.4 (95% confidence interval [CI] = 0.9 to 2.3), and 0% (0 of 31), respectively, for those with unsatisfactory aspirate specimens and 8.2% (148 of 1816), with RR = 1.6 (95% CI = 1.1 to 2.3), and 3.1% (25 of 811), with RR = 1.2 (95% CI = 0.8 to 2.0), respectively, for those with normal cytology in aspirates. Compared with women from whom no fluid was obtained, incidences and adjusted RRs for women in group 1 with epithelial hyperplasia and atypical hyperplasia in aspirates were 10.8% (52 of 483), with RR = 2.4 (95% CI = 1.6 to 3.7), and 13.8% (12 of 87), with RR = 2.8 (95% CI = 1.5 to 5.5), respectively, while those for women in group 2 were 5.5% (25 of 457) and 0% (0 of 22), respectively, with a combined RR = 2.0 (95% CI = 1.3 to 3.3). CONCLUSION: The results obtained with the newly followed women independently confirmed previous findings that women with abnormal cytology in nipple aspirates of breast fluid have an increased risk of breast cancer. PMID- 11734596 TI - Hormone replacement therapy and colorectal adenoma recurrence among women in the Polyp Prevention Trial. AB - BACKGROUND: Epidemiologic studies have suggested that estrogen may protect against the development of colorectal cancers and adenomatous polyps. We conducted a prospective study to evaluate the association between hormone replacement therapy (HRT) and adenoma recurrence among perimenopausal and postmenopausal women participating in the Polyp Prevention Trial, a randomized dietary intervention study of individuals with colorectal adenomas. METHODS: We used a questionnaire and interviews to collect detailed information, at baseline and at each of four annual study visits, from 620 women regarding hormone use, menopausal status, diet, alcohol consumption, and other risk factors. Adenoma recurrence was ascertained by complete colonoscopy at baseline and after 1 and 4 years. Logistic regression models were used to evaluate the association between hormone use and adenoma recurrence after adjusting for intervention group and for age and body mass index at baseline. All statistical tests were two-sided. RESULTS: Adenomas recurred in 200 women. There was no overall association between adenoma recurrence and either overall hormone use (odds ratio [OR] = 1.01; 95% confidence interval [CI] = 0.70 to 1.45), combined estrogen and progestin use (OR = 0.94; 95% CI = 0.57 to 1.56), or unopposed estrogen use (OR = 1.04; 95% CI = 0.68 to 1.59). HRT use was associated with a reduction in risk for recurrence of distal adenomas (OR = 0.56; 95% CI = 0.32 to 1.00) and a statistically nonsignificant increase in risk for recurrence of proximal adenomas (OR = 1.39; 95% CI = 0.85 to 2.26). We observed a statistically significant interaction between the HRT-adenoma recurrence association and age (P =.02). HRT was associated with a 40% reduced risk of adenoma recurrence among women older than 62 years (OR = 0.58; 95% CI = 0.35 to 0.97) but with an increased risk among women younger than 62 years (OR = 1.99; 95% CI = 1.11 to 3.55). CONCLUSIONS: HRT was not associated with a reduced risk for overall adenoma recurrence in this trial cohort, although there was a suggestion of an age interaction. The effect of age on the association needs to be confirmed in other adenoma recurrence trials. PMID- 11734597 TI - Risk of pneumonitis in breast cancer patients treated with radiation therapy and combination chemotherapy with paclitaxel. AB - BACKGROUND: Some chemotherapy (CT) drugs, including taxanes, may enhance the effectiveness of radiation therapy (RT). However, combining these therapies may increase the incidence of radiation pneumonitis, a lung inflammation. In a retrospective cohort study, we evaluated the incidence of radiation pneumonitis in breast cancer patients treated with RT and standard adjuvant CT by use of doxorubicin (Adriamycin) and cyclophosphamide, with and without paclitaxel. METHODS: Forty-one patients with breast cancer were treated with RT and adjuvant CT, including paclitaxel. Paclitaxel and RT (to breast-chest wall in all and lymph nodes in some) were delivered sequentially in 20 patients and concurrently in 21 patients. Paclitaxel was given weekly in some patients and every 3 weeks in other patients. The incidence of radiation pneumonitis was compared with that among patients in our database whose treatments did not include paclitaxel (n = 1286). The percentage of the lung volume irradiated was estimated. The Cox proportional hazards model was used to find covariates that may be associated with the observed outcomes. All P values were two-sided. RESULTS: Radiation pneumonitis developed in six of the 41 patients. Three patients received paclitaxel concurrently with RT, and three received it sequentially (P =.95). The mean percentage of lung volume irradiated was 20% in patients who developed radiation pneumonitis and 22% in those who did not (P =.6). For patients treated with CT including paclitaxel, the crude rate of developing radiation pneumonitis was 14.6% (95% confidence interval [CI] = 5.6% to 29.2%). For patients treated with CT without paclitaxel, the crude rate of pneumonitis was 1.1% (95% CI = 0.2% to 2.3%). The difference between the crude rates with or without paclitaxel is highly statistically significant (P<.0001). The mean time to develop radiation pneumonitis in patients treated concurrently with RT and paclitaxel was statistically significantly shorter in patients receiving paclitaxel weekly than in those receiving it every 3 weeks (P =.002). CONCLUSIONS: The use of paclitaxel and RT in the primary treatment of breast cancer should be undertaken with caution. Clinical trials with the use of combination CT, including paclitaxel plus RT, whether concurrent or sequential, must evaluate carefully the incidence of radiation pneumonitis. PMID- 11734598 TI - Breast carcinoma in situ: risk factors and screening patterns. AB - BACKGROUND: Risk factors associated with invasive breast cancer are well documented, but those associated with breast carcinoma in situ are not well defined. METHODS: We conducted a population-based, case-control study among female residents of Connecticut to identify risk factors for breast carcinoma in situ. Case patients, diagnosed with ductal carcinoma in situ (DCIS) (n = 875) or lobular carcinoma in situ (LCIS) (n = 123), were matched by 5-year age groups with control subjects (n = 999). Case patients were diagnosed between September 15, 1994, through March 14, 1998, and all subjects were between the ages of 20 and 79 years. Information on risk factors and cancer-screening history was collected by telephone interviews. Conditional logistic regression was used to determine odds ratios (ORs) for the association of these factors with the risk of DCIS and LCIS. RESULTS: Case patients with DCIS were more likely than control subjects to report a family history of breast cancer (OR = 1.48; 95% confidence interval [CI] = 1.19 to 1.85) or previous breast biopsy (OR = 3.56; 95% CI = 2.86 to 4.43). They also had fewer full-term pregnancies (OR = 0.86; 95% CI = 0.80 to 0.93) and were older at first full-term pregnancy (OR for being 20-29 years old relative to being <20 years old = 1.68; 95% CI = 1.17 to 2.43) and at menopause (OR for being > or =55 years old relative to being <45 years old = 1.71; 95% CI = 1.05 to 2.77). DCIS case patients were more likely than control subjects to have had a mammographic examination (OR = 2.46; 95% CI = 1.78 to 3.40) or an annual clinical breast examination (OR = 1.83; 95% CI = 1.48 to 2.26). DCIS patients and control subjects did not differ with respect to oral contraceptive use, hormone replacement therapy, alcohol consumption or smoking history, or breast self examination. Associations for LCIS were similar. CONCLUSIONS: The risk factors associated with DCIS and LCIS are similar to those associated with invasive breast cancer. Diagnosis of DCIS is associated with increased mammography screening. PMID- 11734599 TI - Manganese superoxide dismutase alanine-to-valine polymorphism at codon 16 and lung cancer risk. PMID- 11734600 TI - Determining cause of death in prostate cancer: are death certificates valid? PMID- 11734601 TI - Leptin enhances NMDA receptor function and modulates hippocampal synaptic plasticity. AB - The obese gene product leptin is an important signaling protein that regulates food intake and body weight via activation of the hypothalamic leptin receptor (Ob-Rb; Jacob et al., 1997). However, there is growing evidence that Ob-Rb is also expressed in CNS regions, not directly associated with energy homeostasis (Mercer et al., 1996; Hakansson et al., 1998). In the hippocampus, an area of the brain involved in learning and memory, we have found that leptin facilitates the induction of synaptic plasticity. Leptin converts short-term potentiation of synaptic transmission induced by primed burst stimulation of the Schaffer collateral commissural pathway into long-term potentiation. The mechanism underlying this effect involves facilitation of NMDA receptor function because leptin rapidly enhances NMDA-induced increases in intracellular Ca(2+) levels ([Ca(2+)](i)) and facilitates NMDA, but not AMPA, receptor-mediated synaptic transmission. The signaling mechanism underlying these effects involves activation of phosphoinositide 3-kinase, mitogen-activated protein kinase, and Src tyrosine kinases. These data indicate that a novel action of leptin in the CNS is to facilitate hippocampal synaptic plasticity via enhanced NMDA receptor mediated Ca(2+) influx. Impairment of this process may contribute to the cognitive deficits associated with diabetes mellitus. PMID- 11734602 TI - NudC associates with Lis1 and the dynein motor at the leading pole of neurons. AB - NUDC is a highly conserved protein important for nuclear migration and viability in Aspergillus nidulans. Mammalian NudC interacts with Lis1, a neuronal migration protein important during neocorticogenesis, suggesting a conserved mechanism of nuclear movement in A. nidulans and neuronal migration in the developing mammalian brain (S. M. Morris et al., 1998). To further investigate this possibility, we show for the first time that NudC, Lis1, and cytoplasmic dynein intermediate chain (CDIC) colocalize at the microtubule organizing center (MTOC) around the nucleus in a polarized manner facing the leading pole of cerebellar granule cells with a migratory morphology. In neurons with stationary morphology, NudC is distributed throughout the soma and colocalizes with CDIC and tubulin in neurites as well as at the MTOC. At the subcellular level, NudC, CDIC, and p150 dynactin colocalize to the interphase microtubule array and the MTOC in fibroblasts. The observed colocalization is confirmed biochemically by coimmunoprecipitation of NudC with CDIC and cytoplasmic dynein heavy chain (CDHC) from mouse brain extracts. Consistent with its expression in individual neurons, a high level of NudC is detected in regions of the embryonic neocortex undergoing extensive neurogenesis as well as neuronal migration. These data suggest a biochemical and functional interaction of NudC with Lis1 and the dynein motor complex during neuronal migration in vivo. PMID- 11734603 TI - Metabotropic glutamate receptors drive the endocannabinoid system in hippocampus. AB - Endocannabinoids are key intercellular signaling molecules in the brain, but the physiological regulation of the endocannabinoid system is not understood. We used the retrograde signal process called depolarization-induced suppression of inhibition (DSI) to study the regulation of this system. DSI is produced when an endocannabinoid released from pyramidal cells suppresses IPSCs by activating CB1R cannabinoid receptors located on inhibitory interneurons. We now report that activation of group I metabotropic glutamate receptors (mGluRs) enhances DSI and that this effect is blocked by antagonists of both mGluRs and of CB1R. We also found that DSI is absent in CB1R knock-out (CB1R(-/-)) mice, and, strikingly, that mGluR agonists have no effect on IPSCs in these mice. We conclude that group I mGluR-induced enhancement of DSI, and suppression of IPSCs, is actually mediated by endocannabinoids. This surprising result opens up new approaches to the investigation of cannabinoid actions in the brain. PMID- 11734604 TI - Binding characteristics of radiofluorinated 6-dialkylamino-2-naphthylethylidene derivatives as positron emission tomography imaging probes for beta-amyloid plaques in Alzheimer's disease. AB - Senile plaques (SPs) and neurofibrillary tangles (NFTs) are hallmark pathologies accompanying the neurodegeneration involved in Alzheimer's disease (AD), for which beta-amyloid (Abeta) peptide is a major constituent of SPs. Our laboratories previously developed the hydrophobic, fluorescent molecular-imaging probe 2-(1-(6-[(2-[(18)F]fluoroethyl)(methyl)amino]-2 naphthyl)ethylidene)malononitrile ([(18)F]FDDNP), which crosses the blood-brain barrier and determines the localization and load of SPs and NFTs in vivo in AD patients. In this report, we used fluorimetric and radioactive binding assays to determine the binding affinities of FDDNP and its analog, 1-(6-[(2 [(18)F]fluoroethyl)(methyl)amino]naphthalen-2-yl)ethanone ([(18)F]FENE), to synthetic fibrils of Abeta(1-40). FDDNP and FENE both appeared to bind to two kinetically distinguishable binding sites on Abeta(1-40) fibrils. Fluorescence titrations yielded apparent K(d) values of 0.12 and 0.16 nm for high-affinity binding sites for FDDNP and FENE, respectively, and apparent K(d) values of 1.86 and 71.2 nm for the low-affinity binding sites. The traditional radioactive binding assays also produced apparent K(d) values in the low nanomolar range. The presence of two kinetically distinguishable binding sites for FDDNP and FENE suggests multiple binding sites for SPs and identifies the parameters that allow for the structural optimization of this family of probes for in vivo use. The high-affinity binding of the probes to multiple binding sites on fibrils are consistent with results obtained with digital autoradiography, immunohistochemistry, and confocal fluorescence microscopy using human brain specimens of AD patients. PMID- 11734605 TI - Reduction of peak pressure on the forefoot with a rigid rocker-bottom postoperative shoe. AB - An ideal postoperative shoe should be comfortable to wear and protect the foot during recovery from surgery or injury. Protection is assumed to be related to peak pressure and force-time integral under the foot. This study compared a commonly used postoperative shoe with a new postoperative shoe that incorporates a rigid sole with an 11 degrees rocker bottom. The new postoperative shoe significantly reduced peak pressures on the forefoot by 20%. The amount of pressure and force-time integral reduction is compared with other values found in the literature for various shoe modifications. Indications for rocker-bottom shoes are also explored. PMID- 11734606 TI - Evaluation of pressure threshold prior to foot ulceration: one-versus two-point static touch. AB - A prospective study of 29 patients with diabetic neuropathy and 47 nondiabetic patients with tarsal tunnel syndrome were evaluated with computer-assisted neurosensory testing at three sites on the foot. The sensitivity and specificity of one-point static touch thresholds for identifying the presence of large fiber axonal loss was done using the calculated thresholds for monofilaments derived from their markings. The sensitivity for one-point static touch in identifying axonal loss was 33% for the 5.07, 38% for the 4.93, 50% for the 4.17, and 60% for the 4.08 monofilament-equivalent, with a specificity of 100% at each level. Therefore, one-point static touch testing, even using monofilaments thinner than 5.07, has a high percentage of false-negative results in identifying patients with axonal loss. PMID- 11734607 TI - The effect of customized insoles on the reduction of postwork discomfort. AB - This study examined the effect of customized insoles in relieving postwork discomfort in healthy individuals whose jobs require long periods of standing and walking. CompuSole insoles were worn by 122 New York City Police Department officers for up to 5 weeks for an average of 7 hours per day. The officers walked an average of 3 miles per day. Before the study, one-fifth of the police officers in this study experienced foot pain or discomfort at the end of their workday; 15% had calluses, corns, or athlete's foot; 18% had sought treatment for a foot problem in the past; and 20% had worn foot orthoses. There was a significant reduction in tiredness in the feet at the end of the day after wearing the insoles, but no improvement in back or leg discomfort. At the end of the workday, 68% had less foot discomfort and 60% were more comfortable at work when wearing the insoles. PMID- 11734609 TI - Survey of the effects of aerobic dance on the lower extremity in aerobic instructors. AB - The rate of aerobic dance injuries has been high for two decades. To determine the types of lower-extremity injuries to aerobic instructors, a questionnaire was sent to 18 fitness centers in the Sydney, Australia, metropolitan area requesting information on the number and types of injuries, frequency of activity levels, footwear worn, and treatments sought. The reported rate of injury was 77%. The leg was the most common site of injury, reported by 52.9% of respondents, followed by the foot and ankle (32.8%), and the knee (20%). These figures are comparable to previous studies. Further investigation is warranted into causes and preventive measures, and information on the kinetics and kinematics of the lower extremity may increase understanding of the incidence of lower-extremity injuries to aerobic instructors and participants. PMID- 11734608 TI - Patient satisfaction with oral versus nonoral therapeutic approaches in onychomycosis. AB - The follow-up results of a 9-month observational study of 150 onychomycosis patients treated with a variety of mechanical, topical, and oral therapies by podiatric physicians and dermatologists are presented. Changes from baseline in toenail condition and patient satisfaction were assessed at 4- and 9-month follow up. At 9 months, patients who had received oral therapy reported significantly fewer onychomycosis-related problems in social situations, including embarrassment or self-consciousness about the appearance of nails, avoidance of contact by others, being perceived as unclean or untidy, and the desire to keep their nails concealed. Patient-reported satisfaction with the treatment program was significantly higher for those receiving oral therapy than for those receiving nonoral therapy. PMID- 11734610 TI - The role of revascularization in transmetatarsal amputations. AB - Data from 37 patients who underwent a transmetatarsal amputation from January 1993 to April 1996 were reviewed. The mean age and diabetes duration of the subjects were 54.9 (+/- 13.2) years and 16.6 (+/- 8.9) years, respectively. The follow-up period averaged 42.1 (+/- 11.2) months. At the time of follow-up, 29 (78.4%) of the 37 patients still had foot salvage, 8 (21.6%) had progressed to below-the-knee amputation, and 15 (40.5%) had undergone lower-extremity revascularization. Twelve (80%) of the 15 revascularized patients preserved their transmetatarsal amputation level at a follow-up of 36.4 months. The authors concluded that at a maximum of 3 years follow-up after initial amputation, transmetatarsal amputation was a successful amputation level. PMID- 11734611 TI - Internal fixation of a displaced tibial sesamoid fracture. AB - The authors present a surgical technique for the preservation and repair of an acutely fractured sesamoid using internal fixation of the sesamoid. A case report demonstrating the technique for the open reduction and internal fixation of a fractured tibial sesamoid is presented. The authors recommend this procedure as a viable alternative to surgical excision of the tibial sesamoid. The use of the procedure as an adjunct for the surgical treatment of recalcitrant traumatic sesamoiditis is also discussed. PMID- 11734612 TI - Terbinafine-associated taste disturbance with normal taste threshold scores. AB - The medical literature reveals numerous reports of transient taste disturbance associated with the use of oral terbinafine. A review of these reports, however, fails to confirm taste disturbances by formal taste testing. In this article, a case of long-standing taste dysfunction in a patient who exhibited normal formal taste thresholds is described. PMID- 11734613 TI - Efficacy of terbinafine for toenail onychomycosis. PMID- 11734615 TI - Neurotensin and dopamine interactions. AB - Interactions between the classical monoamine neurotransmitter dopamine (DA) and the peptide neurotransmitter neurotensin (NT) in the central nervous system (CNS) have now been investigated for over two decades. Interest in this topic has been sustained, primarily because of the potential clinical relevance of these interactions to schizophrenia and drug abuse. In the past five years, important new discoveries in the NT field have markedly expanded our previous database. Additional NT receptors have been cloned, and novel and refined techniques have contributed to a more detailed description of the anatomy of the CNS NT system. Additionally, lipophilic NT receptor antagonists, active in the CNS after peripheral administration, have rendered more facile the investigation of the physiologic importance of endogenous NT at electrophysiologic, neurochemical, and behavioral levels. In the present review, the discussion of NT/DA interactions will progress from a discussion of the anatomical interactions between these two systems, to electrophysiologic and neurochemical interactions, and finally to behavioral implications-always with focus toward the potential clinical relevance of the data. The discussion of interactions between NT and DA systems will be limited to those occurring within the CNS. Moreover, because the DA projections from the midbrain to the striatum account for the bulk of the DA innervation in the CNS, we will focus on NT/DA interactions within these brain regions. Last, because of the extensive literature on NT/DA interactions available in the rat, our discussion will be based primarily on studies using this species. PMID- 11734616 TI - Norepinephrine and beta 2-adrenergic receptor stimulation regulate CD4+ T and B lymphocyte function in vitro and in vivo. AB - Historically, norepinephrine and the sympathetic nervous system have been associated with the "fight or flight" response, and they also contribute to the regulation of autonomic activity within the body, such as cardiovascular function. In addition, evidence over the past 30 years suggests that norepinephrine may also regulate the function of immune cells that protect the body against pathogens. The presence of sympathetic nerve fibers and the release of norepinephrine within lymphoid organs represent a mechanism by which signals from the central nervous system may influence immune cell function. The T cell dependent antibody response is essential to successful host defense against numerous environmental pathogens. It is during this response that CD4+ T and B lymphocytes are activated to produce cytokines and antibody, respectively, leading to immune competence and protection. The goal of this review is to discuss the evidence supporting the release of norepinephrine within lymphoid organs and the expression of the beta2-adrenergic receptor by CD4+ T and B lymphocytes. We also discuss the mechanisms by which beta2-adrenergic receptor stimulation affects the level of cytokine and antibody produced by these cells both in vitro and in vivo. In cases where conflicting findings have been reported, we discuss potential variables that may have contributed to these conflicting findings. To conclude, we discuss the disease- and health-specific implications of the basic research being done in the area of sympathetic nervous system regulation of T and B lymphocyte function. PMID- 11734617 TI - International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors. AB - Four adenosine receptors have been cloned and characterized from several mammalian species. The receptors are named adenosine A(1), A(2A), A(2B), and A(3). The A(2A) and A(2B) receptors preferably interact with members of the G(s) family of G proteins and the A(1) and A(3) receptors with G(i/o) proteins. However, other G protein interactions have also been described. Adenosine is the preferred endogenous agonist at all these receptors, but inosine can also activate the A(3) receptor. The levels of adenosine seen under basal conditions are sufficient to cause some activation of all the receptors, at least where they are abundantly expressed. Adenosine levels during, e.g., ischemia can activate all receptors even when expressed in low abundance. Accordingly, experiments with receptor antagonists and mice with targeted disruption of adenosine A(1), A(2A), and A(3) expression reveal roles for these receptors under physiological and particularly pathophysiological conditions. There are pharmacological tools that can be used to classify A(1), A(2A), and A(3) receptors but few drugs that interact selectively with A(2B) receptors. Testable models of the interaction of these drugs with their receptors have been generated by site-directed mutagenesis and homology-based modelling. Both agonists and antagonists are being developed as potential drugs. PMID- 11734618 TI - P2X receptors and nociception. AB - The potential importance for nociception of P2X receptors, the ionotropic receptors activated by ATP, is underscored by the variety of pain states in which this endogenous ligand can be released. Several important findings have been made recently indicating that P2X receptors can be involved in pain mechanisms both centrally and in the periphery. The roles of ATP at these two sites and the P2X receptor subtypes involved appear to be different. In the periphery, ATP can be released as a result of tissue injury, visceral distension, or sympathetic activation and can excite nociceptive primary afferents by acting at homomeric P2X(3) or heteromeric P2X(2/3) receptors. Centrally, ATP released from central afferent terminals or second order neurons can modulate neurotransmitter release or postsynaptically activate neurons involved in central nociceptive transmission, with P2X(2), P2X(4), P2X(6), and some other receptors being potentially involved. Evidence from in vivo studies suggests that peripheral ATPergic mechanisms are most important under conditions of acute tissue injury and inflammation whereas the relevance of central mechanisms appears to be more limited. Furthermore, the release of ATP and P2X receptor-mediated afferent activation appear to have been implicated in visceral and neuropathic pain; the importance of the ATPergic component in these states needs to be investigated further. Thus, peripheral P2X receptors, and homomeric P2X(3) and/or heteromeric P2X(2/3) receptors in particular, constitute attractive targets for analgesic drugs. The development of selective antagonists of these receptors, suitable for a systemic in vivo use although apparently difficult, may prove a useful strategy to generate analgesics with a novel mechanism of action. PMID- 11734619 TI - Drug transporters in the central nervous system: brain barriers and brain parenchyma considerations. AB - Drug transport in the central nervous system is highly regulated not only by the blood-brain and the blood-cerebrospinal fluid barriers but also in brain parenchyma. The novel localization of drug transporters in brain parenchyma cells, such as microglia and astrocytes, suggest a reconsideration of the present conceptualization of brain barriers as it relates to drug transport. That is, the cellular membranes of parenchyma cells act as a second "barrier" to drug permeability and express transporters whose properties appear similar to those localized at the conventional brain barriers. This review will focus on the molecular characteristics, localization, and substrate specificities of several classes of well known membrane drug transporters (i.e., the organic cation, organic anion, nucleoside, P-glycoprotein, and multidrug resistance proteins) in the brain. Comparisons to similar transporters localized within the peripheral system and clinical implications of the functional expression of specific drug transport families will be discussed when appropriate. Nutrient and neurotransmitter transporters, whose characteristics have been reviewed extensively elsewhere, will not be considered in this review. PMID- 11734620 TI - Animal models of nociception. AB - The study of pain in awake animals raises ethical, philosophical, and technical problems. We review the ethical standards for studying pain in animals and emphasize that there are scientific as well as moral reasons for keeping to them. Philosophically, there is the problem that pain cannot be monitored directly in animals but can only be estimated by examining their responses to nociceptive stimuli; however, such responses do not necessarily mean that there is a concomitant sensation. The types of nociceptive stimuli (electrical, thermal, mechanical, or chemical) that have been used in different pain models are reviewed with the conclusion that none is ideal, although chemical stimuli probably most closely mimic acute clinical pain. The monitored reactions are almost always motor responses ranging from spinal reflexes to complex behaviors. Most have the weakness that they may be associated with, or modulated by, other physiological functions. The main tests are critically reviewed in terms of their sensitivity, specificity, and predictiveness. Weaknesses are highlighted, including 1) that in most tests responses are monitored around a nociceptive threshold, whereas clinical pain is almost always more severe; 2) differences in the fashion whereby responses are evoked from healthy and inflamed tissues; and 3) problems in assessing threshold responses to stimuli, which continue to increase in intensity. It is concluded that although the neural basis of the most used tests is poorly understood, their use will be more profitable if pain is considered within, rather than apart from, the body's homeostatic mechanisms. PMID- 11734621 TI - Estrogen receptors and proliferation markers in primary and recurrent breast cancer. AB - To elucidate the clinical importance of estrogen receptor (ER) beta in breast cancer, 29 archival primary breast cancer specimens, six locally recurrent cancers, and five benign mammary tumors were examined histochemically for ERalpha, ERbeta and the proliferation markers Ki67 and cyclin A. In benign tumors, most epithelial cells contained ERbeta, but ERalpha was rare. In primary cancers, both ERalpha and ERbeta occurred in epithelial cells, the presence of ERbeta being associated with elevated expression of Ki67 and cyclin A, and ERalpha with decreased levels. Thus, the highest content of proliferation markers was seen in primary cancers that were ERalpha(-) ERbeta(+). Most Ki67-containing cells coexpressed ERbeta, but few showed ERalpha. In locally recurring cancers, ERalpha, ERbeta, and Ki67 were more highly expressed than in the corresponding primary tumors, and many cells containing ERbeta, but few with ERalpha, expressed Ki67. Surprisingly, ERbeta, but not ERalpha, was seen in the stromal cells of both primary and recurrent cancers. Because the response of breast cancers to tamoxifen therapy is correlated with the presence of ERalpha, cancer cells that lack ERalpha but contain ERbeta and proliferation markers represent a novel population of apparently proliferating cells that probably are not targeted by the current antiestrogens. Thus, appropriate ERbeta-specific ligands, perhaps in combination with tamoxifen, may be useful in improving the treatment of breast cancers. PMID- 11734622 TI - Epstein-Barr virus nuclear antigen 1 activates transcription from episomal but not integrated DNA and does not alter lymphocyte growth. AB - By binding to a cis-acting element (oriP) in the Epstein-Barr virus (EBV) genome, EBV nuclear antigen 1 (EBNA1) enables persistence and enhances transcription from EBV episomes. To investigate whether EBNA1 also directly affects cell gene transcription, we conditionally expressed a Flag-tagged dominant negative EBNA1 (FDNE) in an EBV immortalized lymphoblastoid cell line, in which the EBV genome is integrated into cell DNA. FDNE induction inhibited expression from an EBNA1 dependent oriP reporter plasmid by more than 90% in these cells but did not affect expression from integrated EBV or oriP reporter DNA. FDNE induction also did not alter expression of more than 1,800 cellular mRNAs. Lymphoblastoid cell line growth under a variety of conditions was unaffected by FDNE induction. Although Gal4-VP16 and EBNA1 strongly activated and coactivated a Gal4-VP16- and oriP-dependent promoter that was on an episome, only Gal4-VP16 activated the promoter when it was integrated into chromosomal DNA. These data indicate that EBNA1 is specifically deficient in activation of an integrated oriP enhancer and does not affect cell growth or gene expression through an interaction with cognate chromosomal DNA. PMID- 11734623 TI - Inadequate lung development and bronchial hyperplasia in mice with a targeted deletion in the Dutt1/Robo1 gene. AB - Chromosome 3 allele loss in preinvasive bronchial abnormalities and carcinogen exposed, histologically normal bronchial epithelium indicates that it is an early, possibly the first, somatic genetic change in lung tumor development. Candidate tumor suppressor genes have been isolated from within distinct 3p regions implicated by heterozygous and homozygous allele loss. We have proposed that DUTT1, nested within homozygously deleted regions at 3p12-13, is the tumor suppressor gene that deletion-mapping and tumor suppression assays indicate is located in proximal 3p. The same gene, ROBO1 (accession number ), was independently isolated as the human homologue of the Drosophila gene, Roundabout. The gene, coding for a receptor with a domain structure of the neural-cell adhesion molecule family, is widely expressed and has been implicated in the guidance and migration of axons, myoblasts, and leukocytes in vertebrates. A deleted form of the gene, which mimics a naturally occurring, tumor-associated human homozygous deletion of exon 2 of DUTT1/ROBO1, was introduced into the mouse germ line. Mice homozygous for this targeted mutation, which eliminates the first Ig domain of Dutt1/Robo1, frequently die at birth of respiratory failure because of delayed lung maturation. Lungs from these mice have reduced air spaces and increased mesenchyme, features that are present some days before birth. Survivors acquire extensive bronchial epithelial abnormalities including hyperplasia, providing evidence of a functional relationship between a 3p gene and the development of bronchial abnormalities associated with early lung cancer. PMID- 11734624 TI - PAS-A domain of phosphorelay sensor kinase A: a catalytic ATP-binding domain involved in the initiation of development in Bacillus subtilis. AB - The major sensor kinase controlling the initiation of development in Bacillus subtilis, KinA, functions by activating the phosphorelay signal-transduction system in response to as yet unknown signal ligands. KinA contains, within its amino-terminal signal-sensing region, three PAS domains that, in other proteins, are known to be involved in sensing changes in oxygen concentration and redox potential among other functions. The most amino-terminal PAS domain, PAS-A, was found to bind ATP and catalyze exchange of phosphate between ATP and nucleoside diphosphates. A cysteine-to-alanine mutation in PAS-A increased the affinity for ATP 5-fold, decreased the exchange reaction 2-fold, and stimulated KinA-dependent sporulation. A model for the role of ATP and the exchange reaction in the PAS domain in sensor kinase signal transduction is presented in which the free energy of nucleotide hydrolysis drives the conformational changes that activate or deactivate the sensor kinase in response to signal ligand binding. PMID- 11734626 TI - Calcium influx selects the fast mode of endocytosis in the synaptic terminal of retinal bipolar cells. AB - To investigate the regulation of endocytosis by Ca(2+), we have made capacitance measurements in the synaptic terminal of depolarizing bipolar cells from the retina of goldfish. After a brief depolarization, all of the excess membrane was retrieved rapidly (tau approximately 1 s). But when the rise in free [Ca(2+)] was reduced by the introduction of Ca(2+) buffers [1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetate (BAPTA) or EGTA], a large fraction of the membrane was retrieved by a second, slower mechanism (tau > or = 10 s). The block of fast endocytosis by EGTA could be overcome by increasing the amplitude of the Ca(2+) current, demonstrating that Ca(2+) influx was the trigger for fast endocytosis. These manipulations of the Ca(2+) signal altered the relative proportions of fast and slow endocytosis but did not modulate the rate constants of these processes. A brief stimulus that triggered fast endocytosis did not generate a significant rise in the spatially averaged [Ca(2+)], indicating that Ca(2+) regulated endocytosis through an action close to the active zone. The slow mode of retrieval occurred at the resting [Ca(2+)]. These results demonstrate that Ca(2+) influx couples fast endocytosis and exocytosis at this synapse. PMID- 11734625 TI - Doppel-induced cerebellar degeneration in transgenic mice. AB - Doppel (Dpl) is a paralog of the mammalian prion protein (PrP); it is abundant in testes but expressed at low levels in the adult central nervous system. In two Prnp-deficient (Prnp(0/0)) mouse lines (Ngsk and Rcm0), Dpl overexpression correlated with ataxia and death of cerebellar neurons. To determine whether Dpl overexpression, rather than the dysregulation of genes neighboring the Prn gene complex, was responsible for the ataxic syndrome, we placed the mouse Dpl coding sequence under the control of the Prnp promoter and produced transgenic (Tg) mice on the Prnp(0/0)-ZrchI background (hereafter referred to as ZrchI). ZrchI mice exhibit neither Dpl overexpression nor cerebellar degeneration. In contrast, Tg(Dpl)ZrchI mice showed cerebellar granule and Purkinje cell loss; the age of onset of ataxia was inversely proportional to the levels of Dpl protein. Crosses of Tg mice overexpressing wild-type PrP with two lines of Tg(Dpl)ZrchI mice resulted in a phenotypic rescue of the ataxic syndrome, while Dpl overexpression was unchanged. Restoration of PrP expression also rendered the Tg(Dpl) mice susceptible to prion infection, with incubation times indistinguishable from non Tg controls. Whereas the rescue of Dpl-induced neurotoxicity by coexpression of PrP argues for an interaction between the PrP and Dpl proteins in vivo, the unaltered incubation times in Tg mice overexpressing Dpl in the central nervous system suggest that Dpl is unlikely to be involved in prion formation. PMID- 11734627 TI - The role of the third beta strand in gp120 conformation and neutralization sensitivity of the HIV-1 primary isolate DH012. AB - Neutralization of HIV-1 primary isolates has been a tremendous challenge for AIDS vaccine development. Here, we identify a single amino acid change (T198P) in gp120 that alters the neutralization sensitivity of the primary isolate DH012 to antibodies against multiple neutralization epitopes that include the V3, CD4 induced, and CD4 binding sites in gp120. This mutation is located in the V1/V2 stem region that forms the third beta strand (beta3) of the bridging sheet of gp120. The conformation of variable loops, especially V1/V2 and V3, was proposed to regulate the accessibility of these neutralization epitopes. The results of this study indicate a direct association between the V1/V2 and V3 loops of DH012 gp120. The single amino acid mutation T198P in the beta3 severely compromises the interaction between the V1/V2 and V3 loops. These results suggest that interaction of V1/V2 and V3 can mask the neutralization epitopes and that the beta3 plays a critical role in determining the neutralization sensitivity by modulating the interaction. This study provides an insight into why primary isolates are relatively resistant to antibody neutralization and might facilitate the development of anti-HIV strategies against HIV-1 infection. PMID- 11734628 TI - The coxsackievirus and adenovirus receptor is a transmembrane component of the tight junction. AB - The coxsackievirus and adenovirus receptor (CAR) mediates viral attachment and infection, but its physiologic functions have not been described. In nonpolarized cells, CAR localized to homotypic intercellular contacts, mediated homotypic cell aggregation, and recruited the tight junction protein ZO-1 to sites of cell-cell contact. In polarized epithelial cells, CAR and ZO-1 colocalized to tight junctions and could be coprecipitated from cell lysates. CAR expression led to reduced passage of macromolecules and ions across cell monolayers, and soluble CAR inhibited the formation of functional tight junctions. Virus entry into polarized epithelium required disruption of tight junctions. These results indicate that CAR is a component of the tight junction and of the functional barrier to paracellular solute movement. Sequestration of CAR in tight junctions may limit virus infection across epithelial surfaces. PMID- 11734629 TI - A "master" in base unpairing during isomerization of a promoter upon RNA polymerase binding. AB - Isomerization of a closed to open complex of a promoter upon RNA polymerase binding involves base unpairing at the -10 region. After potassium permanganate sensitivity of unpaired thymine residues, we studied base unpairing at the -10 region during isomerization upon RNA polymerase binding at the P1 and P3 promoters of the gal operon. Substitution of adenine by 2-amino purine (2-AP) at the invariable A small middle dotT base pair at the -11 position of P1 and P3 prevented unpairing not only at that position but also at the other downstream positions, suggesting a "master" role of the adenine base at -11 of the template strand in overall base unpairing. 2-AP at -11 did not inhibit the formation of RNA polymerase small middle dotpromoter complex and subsequent isomerization of the polymerase. Substitution of adenine by 2-AP at several other positions did not affect thymine unpairing. Changing the position of the amino group from C6 in adenine to C2 in 2-AP is mutational only at the master switch position, -11. PMID- 11734630 TI - Cryoelectron microscopy of lambda phage DNA condensates in vitreous ice: the fine structure of DNA toroids. AB - DNA toroids produced by the condensation of lambda phage DNA with hexammine cobalt (III) have been investigated by cryoelectron microscopy. Image resolution obtained by this technique has allowed unprecedented views of DNA packing within toroidal condensates. Toroids oriented coplanar with the microscope image plane exhibit circular fringes with a repeat spacing of 2.4 nm. For some toroids these fringes are observed around almost the entire circumference of the toroid. However, for most toroids well-defined fringes are limited to less than one-third of the total toroid circumference. Some toroids oriented perpendicular to the image plane reveal DNA polymers organized in a hexagonal close-packed lattice; however, for other toroids alternative packing arrangements are observed. To aid interpretation of electron micrographs, three-dimensional model toroids were generated with perfect hexagonal DNA packing throughout, as well as more physically realistic models that contain crossover points between DNA loops. Simulated transmission electron microscopy images of these model toroids in different orientations faithfully reproduce most features observed in cryoelectron micrographs of actual toroids. PMID- 11734631 TI - Mapping the actin filament with myosin. AB - Structural studies have shown that the heads of the myosin motor molecule bind preferentially to "target zones" of favorably oriented sites on the helices of the actin filament. We present direct evidence for target zones from the interactions of a single myosin head with an actin filament held between two optically trapped beads. With compliant traps, thermal motions of the filament allow the fixed myosin-S1 to interact with at least two zones, observed as a bi modal distribution of filament displacements due to myosin binding, whose spacing is near the 36-nm helix repeat distance. The number of binding events and the "apparent working stroke" (mean displacement with myosin bound), vary periodically as the filament is moved past the fixed myosin by displacing the traps; observed periods are close to 36 nm and the apparent stroke varies from 0 10 nm. We also observe a strong modulation at the 5.5-nm actin monomer repeat confirming that myosin interacts with a single strand and that the actin is not free to rotate. Each interaction can be assigned to an actin monomer and each active zone on the helix is made up of three actin monomers. PMID- 11734632 TI - A targeted dominant negative mutation of the thyroid hormone alpha 1 receptor causes increased mortality, infertility, and dwarfism in mice. AB - Mutations in the thyroid hormone receptor beta (TRbeta) gene result in resistance to thyroid hormone. However, it is unknown whether mutations in the TRalpha gene could lead to a similar disease. To address this question, we prepared mutant mice by targeting mutant thyroid hormone receptor kindred PV (PV) mutation to the TRalpha gene locus by means of homologous recombination (TRalpha1PV mice). The PV mutation was derived from a patient with severe resistance to thyroid hormone that has a frameshift of the C-terminal 14 aa of TRbeta1. We knocked in the same PV mutation to the corresponding TRalpha gene locus to compare the phenotypes of TRalpha1(PV/+) mice with those of TRbeta(PV/+) mice. TRalpha1(PV/+) mice were viable, indicating that the mutation of the TRalpha gene is not embryonic lethal. In drastic contrast to the TRbeta(PV/+) mice, which do not exhibit a growth abnormality, TRalpha1(PV/+) mice were dwarfs. These dwarfs exhibited increased mortality and reduced fertility. In contrast to TRbeta(PV/+) mice, which have a hyperactive thyroid, TRalpha1(PV/+) mice exhibited mild thyroid failure. The in vivo pattern of abnormal regulation of T3 target genes in TRalpha1(PV/+) mice was unique from those of TRbeta(PV/+) mice. The distinct phenotypes exhibited by TRalpha1(PV/+) and TRbeta(PV/+) mice indicate that the in vivo functions of TR mutants are isoform-dependent. The TRalpha1(PV/+) mice may be used as a tool to uncover human diseases associated with mutations in the TRalpha gene and, furthermore, to understand the molecular mechanisms by which TR isoforms exert their biological activities. PMID- 11734633 TI - A genetic screen identifies a cellular regulator of adeno-associated virus. AB - Adeno-associated virus type 2 (AAV2) is a human parvovirus that has attracted attention as a vector for gene transfer. Replication and site-specific integration of the wild-type virus requires binding of the AAV2 Rep proteins to a cis-regulatory element named the Rep recognition sequence (RRS). RRS motifs are found within the cellular AAVS1 integration locus, the viral p5 promoter, and the inverted terminal repeats (ITRs). Here we report the design of a genetic screen based on the yeast one-hybrid assay to identify cellular RRS-binding proteins. We show that the human zinc finger 5 protein (ZF5) binds specifically to RRS motifs in vitro and in vivo. ZF5 is a highly conserved and ubiquitously expressed transcription factor that contains five C-terminal zinc fingers and an N-terminal POZ domain. Ectopic expression of ZF5 leads to an ITR-dependent repression of the autologous p5 promoter and reduces both AAV2 replication and the production of recombinant AAV2. By using deletion and substitution mutants we show that two different domains of ZF5 contribute to AAV2 repression. Negative regulation of the p5 promoter requires the POZ domain, whereas viral replication is inhibited by the zinc finger domain, likely by competing with Rep for binding to the ITR. Identification and characterization of proteins that bind the ITR, the only viral genetic element retained in AAV2 vectors, will lead to new insights into the unique life cycle of AAV2 and will suggest improvements important for its application as a gene therapy vector. PMID- 11734634 TI - Short-cycle structured intermittent treatment of chronic HIV infection with highly active antiretroviral therapy: effects on virologic, immunologic, and toxicity parameters. AB - Although continuous highly active antiretroviral therapy (HAART) is effective for many HIV-infected patients, it can be toxic and prohibitive in cost. By decreasing the total amount of time patients receive medications, intermittent HAART could reduce toxicity and cost. Therefore, we initiated a pilot study in which 10 HIV-infected individuals receiving effective therapy that resulted in levels of HIV RNA <50 copies per ml of plasma and CD4(+) T cell counts >300 cells per mm(3) of whole blood received repeated cycles of 7 days on HAART followed by 7 days off of HAART. Patients maintained suppression of plasma viremia for 32-68 weeks. There was no significant increase in HIV proviral DNA or replication competent HIV in peripheral CD4(+) T cells or HIV RNA in peripheral blood or lymph node mononuclear cells. There was no significant change in CD4(+) T cell counts, no significant increase in CD4(+) or CD8(+) T cells expressing activation markers or producing IFN-gamma in response to HIV, no increase in CD4(+) T cell proliferation to p24 antigen, and no evidence for the development of resistance to HAART medications. There was a significant decrease in serum cholesterol and triglyceride levels. Thus, in this proof-of-concept study, short-cycle intermittent HAART maintained suppression of plasma viremia as well as HIV replication in reservoir sites while preserving CD4(+) T cell counts. In addition, there was a decrease in serum cholesterol and triglyceride levels. Intermittent therapy may be an important strategy to reduce cost and toxicity for HIV-infected individuals. PMID- 11734635 TI - Tox-boxes, fungal secondary metabolites, and plant disease. PMID- 11734636 TI - Dissecting the transcriptional network of pancreatic islets during development and differentiation. PMID- 11734637 TI - Surprises from the airway epithelium. PMID- 11734638 TI - Another protozoan contributes to understanding telomeres and transposable elements. PMID- 11734639 TI - Pattern formation in Escherichia coli: a model for the pole-to-pole oscillations of Min proteins and the localization of the division site. AB - Proper cell division requires an accurate definition of the division plane. In bacteria, this plane is determined by a polymeric ring of the FtsZ protein. The site of Z ring assembly in turn is controlled by the Min system, which suppresses FtsZ polymerization at noncentral membrane sites. The Min proteins in Escherichia coli undergo a highly dynamic localization cycle, during which they oscillate between the membrane of both cell halves. By using computer simulations we show that Min protein dynamics can be described accurately by using the following assumptions: (i) the MinD ATPase self-assembles on the membrane and recruits both MinC, an inhibitor of Z ring formation, and MinE, a protein required for MinC/MinD oscillation, (ii) a local accumulation of MinE is generated by a pattern formation reaction that is based on local self-enhancement and a long range antagonistic effect, and (iii) it displaces MinD from the membrane causing its own local destabilization and shift toward higher MinD concentrations. This local destabilization results in a wave of high MinE concentration traveling from the cell center to a pole, where it disappears. MinD reassembles on the membrane of the other cell half and attracts a new accumulation of MinE, causing a wave like disassembly of MinD again. The result is a pole-to-pole oscillation of MinC/D. On time average, MinC concentration is highest at the poles, forcing FtsZ assembly to the center. The mechanism is self-organizing and does not require any other hypothetical topological determinant. PMID- 11734640 TI - Dimer formation drives the activation of the cell death protease caspase 9. AB - A critical step in the induction of apoptosis is the activation of the apoptotic initiator caspase 9. We show that at its normal physiological concentration, caspase 9 is primarily an inactive monomer (zymogen), and that activity is associated with a dimeric species. At the high concentrations used for crystal formation, caspase 9 is dimeric, and the structure reveals two very different active-site conformations within each dimer. One site closely resembles the catalytically competent sites of other caspases, whereas in the second, expulsion of the "activation loop" disrupts the catalytic machinery. We propose that the inactive domain resembles monomeric caspase 9. Activation is induced by dimerization, with interactions at the dimer interface promoting reorientation of the activation loop. These observations support a model in which recruitment by Apaf-1 creates high local concentrations of caspase 9 to provide a pathway for dimer-induced activation. PMID- 11734641 TI - A second iron-regulatory system in yeast independent of Aft1p. AB - Iron homeostasis in the yeast Saccharomyces cerevisiae is regulated at the transcriptional level by Aft1p, which activates the expression of its target genes in response to low-iron conditions. The yeast genome contains a paralog of AFT1, which has been designated AFT2. To establish whether AFT1 and AFT2 have overlapping functions, a mutant containing a double aft1Deltaaft2Delta deletion was generated. Growth assays established that the single aft2Delta strain exhibited no iron-dependent phenotype. However, the double-mutant aft1Deltaaft2Delta strain was more sensitive to low-iron growth conditions than the single-mutant aft1Delta strain. A mutant allele of AFT2 (AFT2-1(up)), or overexpression of the wild-type AFT2 gene, led to partial complementation of the respiratory-deficient phenotype of the aft1Delta strain. The AFT2-1(up) allele also increased the uptake of (59)Fe in an aft1Delta strain. DNA microarrays were used to identify genes regulated by AFT2. Some of the AFT2-regulated genes are known to be regulated by Aft1p; however, AFT2-1(up)-dependent activation was independent of Aft1p. The kinetics of induction of two genes activated by the AFT2-1(up) allele are consistent with Aft2p acting as a direct transcriptional factor. Truncated forms of Aft1p and Aft2p bound to a DNA duplex containing the Aft1p binding site in vitro. The wild-type allele of AFT2 activated transcription in response to growth under low-iron conditions. Together, these data suggest that yeast has a second regulatory pathway for the iron regulon, with AFT1 and AFT2 playing partially redundant roles. PMID- 11734642 TI - Fast dynamics of halophilic malate dehydrogenase and BSA measured by neutron scattering under various solvent conditions influencing protein stability. AB - Protein thermal dynamics was evaluated by neutron scattering for halophilic malate dehydrogenase from Haloarcula marismortui (HmMalDH) and BSA under different solvent conditions. As a measure of thermal stability in each case, loss of secondary structure temperatures were determined by CD. HmMalDH requires molar salt and has different stability behavior in H(2)O, D(2)O, and in NaCl and KCl solvents. BSA remains soluble in molar NaCl. The neutron experiments provided values of mean-squared atomic fluctuations at the 0.1 ns time scale. Effective force constants, characterizing the mean resilience of the protein structure, were calculated from the variation of the mean-squared fluctuation with temperature. For HmMalDH, resilience increased progressively with increasing stability, from molar NaCl in H(2)O, via molar KCl in D(2)O, to molar NaCl in D(2)O. Surprisingly, however, the opposite was observed for BSA; its resilience is higher in H(2)O where it is less stable than in D(2)O. These results confirmed the complexity of dynamics-stability relationships in different proteins. Softer dynamics for BSA in D(2)O showed that the higher thermostability is associated with entropic fluctuations. In the halophilic protein, higher stability is associated with increased resilience showing the dominance of enthalpic terms arising from bonded interactions. From previous data, it is suggested that these are associated with hydrated ion binding stabilizing the protein in the high-salt solvent. PMID- 11734643 TI - Radiation risk to low fluences of alpha particles may be greater than we thought. AB - Based principally on the cancer incidence found in survivors of the atomic bombs dropped in Hiroshima and Nagasaki, the International Commission on Radiation Protection (ICRP) and the United States National Council on Radiation Protection and Measurements (NCRP) have recommended that estimates of cancer risk for low dose exposure be extrapolated from higher doses by using a linear, no-threshold model. This recommendation is based on the dogma that the DNA of the nucleus is the main target for radiation-induced genotoxicity and, as fewer cells are directly damaged, the deleterious effects of radiation proportionally decline. In this paper, we used a precision microbeam to target an exact fraction (either 100% or < or =20%) of the cells in a confluent population and irradiated their nuclei with exactly one alpha particle each. We found that the frequencies of induced mutations and chromosomal changes in populations where some known fractions of nuclei were hit are consistent with non-hit cells contributing significantly to the response. In fact, irradiation of 10% of a confluent mammalian cell population with a single alpha particle per cell results in a mutant yield similar to that observed when all of the cells in the population are irradiated. This effect was significantly eliminated in cells pretreated with a 1 mM dose of octanol, which inhibits gap junction-mediated intercellular communication, or in cells carrying a dominant negative connexin 43 vector. The data imply that the relevant target for radiation mutagenesis is larger than an individual cell and suggest a need to reconsider the validity of the linear extrapolation in making risk estimates for low dose, high linear-energy-transfer (LET) radiation exposure. PMID- 11734644 TI - Akt/PKB regulates laminin and collagen IV isotypes of the basement membrane. AB - Basement membranes are important for epithelial differentiation, cell survival, and normal and metastatic cell migration. Much is known about their breakdown and remodeling, yet their positive regulation is poorly understood. Our previous analysis of a fibroblast growth factor (FGF) receptor mutation raised the possibility that protein kinase B (Akt/PKB) activated by FGF is connected to the expression of certain laminin and type IV collagen isotypes. Here we test this hypothesis and demonstrate that constitutively active Akt/PKB, an important downstream element of phosphoinositide 3'-kinase signaling, induces the synthesis of laminin-1 and collagen IV isotypes and causes their translocation to the basement membrane. By using promoter-reporter constructs, we show that constitutively active phosphoinositide 3'-kinase-p110 or Akt/PKB activates, whereas dominant negative Akt/PKB inhibits, transcription of laminin beta1 and collagen IV alpha1 in differentiating C2 myoblast- and insulin-induced Chinese hamster ovary-T cell cultures. These results suggest that Akt/PKB activated by receptor tyrosine kinases is involved in the positive regulation of basement membrane formation. The possible role of Akt/PKB-induced laminin and collagen IV synthesis in cell survival and differentiation will be discussed. PMID- 11734645 TI - The LIM domain gene LMO4 inhibits differentiation of mammary epithelial cells in vitro and is overexpressed in breast cancer. AB - LMO4 belongs to a family of LIM-only transcriptional regulators, the first two members of which are oncoproteins in acute T cell leukemia. We have explored a role for LMO4, initially described as a human breast tumor autoantigen, in developing mammary epithelium and breast oncogenesis. Lmo4 was expressed predominantly in the lobuloalveoli of the mammary gland during pregnancy. Consistent with a role in proliferation, forced expression of this gene inhibited differentiation of mammary epithelial cells. Overexpression of LMO4 mRNA was observed in 5 of 10 human breast cancer cell lines. Moreover, in situ hybridization analysis of 177 primary invasive breast carcinomas revealed overexpression of LMO4 in 56% of specimens. Immunohistochemistry confirmed overexpression in a high percentage (62%) of tumors. These studies imply a role for LMO4 in maintaining proliferation of mammary epithelium and suggest that deregulation of this gene may contribute to breast tumorigenesis. PMID- 11734646 TI - Induction of platelet formation from megakaryocytoid cells by nitric oxide. AB - Although the growth factors that regulate megakaryocytopoiesis are well known, the molecular determinants of platelet formation from mature megakaryocytes remain poorly understood. Morphological changes in megakaryocytes associated with platelet formation and removal of senescent megakaryocytes are suggestive of an apoptotic process. Previously, we have established that nitric oxide (NO) can induce apoptosis in megakaryocytoid cell lines. To determine whether there is an association between NO-induced apoptosis and platelet production, we exposed Meg 01 cells to S-nitrosoglutathione (GSNO) with or without thrombopoeitin (TPO) pretreatment and used flow cytometry and electron microscopy to assess platelet sized particle formation. Meg-01 cells treated with TPO alone produced few platelet-sized particles (<3% of total counts), whereas treatment with GSNO alone produced a significant percentage of platelet-sized particles (22 +/- 4% of total counts); when combined with TPO pretreatment, however, GSNO led to a marked increase in platelet-sized particle production (48 +/- 3% of total counts). Electron microscopy confirmed that Meg-01 cells treated with TPO and GSNO yielded platelet-sized particles with morphological features specific for platelet forms. The platelet-sized particle population appears to be functional, because addition of calcium, fibrinogen, and thrombin receptor-activating peptide led to aggregation. These results demonstrate that NO facilitates platelet production, thereby establishing the essential role of NO in megakaryocyte development and thrombopoiesis. PMID- 11734647 TI - Bazooka is required for localization of determinants and controlling proliferation in the sensory organ precursor cell lineage in Drosophila. AB - Asymmetric divisions with two different division orientations follow different polarity cues for the asymmetric segregation of determinants in the sensory organ precursor (SOP) lineage. The first asymmetric division depends on frizzled function and has the mitotic spindle of the pI cell in the epithelium oriented along the anterior-posterior axis, giving rise to pIIa and pIIb, which divide in different orientations. Only the pIIb division resembles neuroblast division in daughter-size asymmetry, spindle orientation along the apical-basal axis, basal Numb localization, and requirement for inscuteable function. Because the PDZ domain protein Bazooka is required for spindle orientation and basal localization of Numb in neuroblasts, we wondered whether Bazooka plays a similar role in the pIIb in the SOP lineage. Surprisingly, Bazooka controls asymmetric localization of the Numb-anchoring protein Pon, but not spindle orientation, in pI and all subsequent divisions. Bazooka also regulates cell proliferation in the SOP lineage; loss of bazooka function results in supernumerary cell divisions and apoptotic cell death. PMID- 11734648 TI - Bazooka and atypical protein kinase C are required to regulate oocyte differentiation in the Drosophila ovary. AB - The par genes, identified by their role in the establishment of anterior posterior polarity in the Caenorhabditis elegans zygote, subsequently have been shown to regulate cellular polarity in diverse cell types by means of an evolutionarily conserved protein complex including PAR-3, PAR-6, and atypical protein kinase C (aPKC). The Drosophila homologs of par-1, par-3 (bazooka, baz), par-6 (DmPar-6), and pkc-3 (Drosophila aPKC, DaPKC) each are known to play conserved roles in the generation of cell polarity in the germ line as well as in epithelial and neural precursor cells within the embryo. In light of this functional conservation, we examined the potential role of baz and DaPKC in the regulation of oocyte polarity. Our analyses reveal germ-line autonomous roles for baz and DaPKC in the establishment of initial anterior-posterior polarity within germ-line cysts and maintenance of oocyte cell fate. Germ-line clonal analyses indicate both proteins are essential for two key aspects of oocyte determination: the posterior translocation of oocyte specification factors and the posterior establishment of the microtubule organizing center within the presumptive oocyte. We demonstrate BAZ and DaPKC colocalize to belt-like structures between germarial cyst cells. However, in contrast to their regulatory relationship in the Drosophila and C. elegans embryos, these proteins are not mutually dependent for their germ-line localization, nor is either protein specifically required for PAR 1 localization to the fusome. Therefore, whereas BAZ, DaPKC, and PAR-1 are functionally conserved in establishing oocyte polarity, the regulatory relationships among these genes are not well conserved, indicating these molecules function differently in different cellular contexts. PMID- 11734649 TI - Three retrotransposon families in the genome of Giardia lamblia: two telomeric, one dead. AB - Transposable elements inhabiting eukaryotic genomes are generally regarded either as selfish DNA, which is selectively neutral to the host organism, or as parasitic DNA, deleterious to the host. Thus far, the only agreed-upon example of beneficial eukaryotic transposons is provided by Drosophila telomere-associated retrotransposons, which transpose directly to the chromosome ends and thereby protect them from degradation. This article reports the transposon content of the genome of the protozoan Giardia lamblia, one of the earliest-branching eukaryotes. A total of three non-long terminal repeat retrotransposon families have been identified, two of which are located at the ends of chromosomes, and the third one contains exclusively dead copies with multiple internal deletions, nucleotide substitutions, and frame shifts. No other reverse transcriptase- or transposase-related sequences were found. Thus, the entire genome of this protozoan, which is not known to reproduce sexually, contains only retrotransposons that are either confined to telomeric regions and possibly beneficial, or inactivated and completely nonfunctional. PMID- 11734650 TI - A likelihood ratio test for evolutionary rate shifts and functional divergence among proteins. AB - Changes in protein function can lead to changes in the selection acting on specific residues. This can often be detected as evolutionary rate changes at the sites in question. A maximum-likelihood method for detecting evolutionary rate shifts at specific protein positions is presented. The method determines significance values of the rate differences to give a sound statistical foundation for the conclusions drawn from the analyses. A statistical test for detecting slowly evolving sites is also described. The methods are applied to a set of Myc proteins for the identification of both conserved sites and those with changing evolutionary rates. Those positions with conserved and changing rates are related to the structures and functions of their proteins. The results are compared with an earlier Bayesian method, thereby highlighting the advantages of the new likelihood ratio tests. PMID- 11734651 TI - Restricted chromosomal silencing in nucleolar dominance. AB - Failure of one parent's chromosomes to organize nucleoli in an interspecific hybrid is an epigenetic phenomenon known as nucleolar dominance. Selective gene silencing on a scale of millions of bp is known to be involved, but the full extent to which nucleolus organizer region (NOR)-bearing chromosomes are inactivated beyond the NORs is unknown. Aided by genome sequence data for Arabidopsis thaliana, we have mapped the extent of nucleolar dominance-induced silencing in Arabidopsis suecica, the allotetraploid hybrid of A. thaliana and Arabidopsis arenosa. Using a sensitive reverse transcription PCR assay, we show that the four A. thaliana NORs, each approximately 4 Mbp in size, are approximately 99.5% silenced in A. suecica vegetative leaves, whereas the NORs inherited from A. arenosa remain fully active. The two A. thaliana NORs, NOR2 and NOR4, abut the telomeres on chromosomes 2 and 4, thus there are no genes distal to the NORs. The three protein-coding genes nearest NOR4 on its centromere proximal side, the closest of which is only 3.1 kb from rRNA gene sequences, are shown to be transcribed in the hybrid despite the silencing of the adjacent approximately 4-Mbp NOR. These data argue against hypotheses in which NOR inactivation is attributed to the spread of silencing from adjacent chromosomal regions, but favor models in which NORs or rRNA genes are the targets of regulation. PMID- 11734652 TI - T cell infiltration of the prostate induced by androgen withdrawal in patients with prostate cancer. AB - Manipulations capable of breaking host tolerance to induce tissue-specific T cell mediated inflammation are of central importance to tumor immunotherapy and our understanding of autoimmunity. We demonstrate that androgen ablative therapy induces profuse T cell infiltration of benign glands and tumors in human prostates. T cell infiltration is readily apparent after 7-28 days of therapy and is comprised predominantly of a response by CD4+ T cells and comparatively fewer CD8+ T cells. Also, T cells within the treated prostate exhibit restricted TCR Vbeta gene usage, consistent with a local oligoclonal response. Recruitment/activation of antigen-presenting cells in treated prostate tissues may contribute to local T cell activation. The induction of T cell infiltration in prostate tissues treated with androgen ablation may have implications for the immunotherapeutic treatment of prostate cancer as well as other hormone-sensitive malignancies, including breast carcinoma. PMID- 11734653 TI - Two-step transcriptional amplification as a method for imaging reporter gene expression using weak promoters. AB - We are developing assays to image tissue-specific reporter gene expression in living mice by using optical methods and positron emission tomography. Approaches for imaging reporter gene expression depend on robust levels of mRNA and reporter protein. Attempts to image reporter gene expression driven by weak promoters are often hampered by the poor transcriptional activity of such promoters. Most tissue-specific promoters are weak relative to stronger but constitutively expressing viral promoters. In this study, we have validated methods to enhance the transcriptional activity of the prostate-specific antigen promoter for imaging by using a two-step transcriptional amplification (TSTA) system. We used the TSTA system to amplify expression of firefly luciferase (fl) and mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) in a prostate cancer cell line (LNCaP). We demonstrate approximately 50-fold (fl) and approximately 12 fold (HSV1-sr39tk) enhancement by using the two-step approach. The TSTA system is observed to retain tissue selectivity. A cooled charge-coupled device optical imaging system was used to visualize the amplified fl expression in living mice implanted with LNCaP cells transfected ex vivo. These imaging experiments reveal a approximately 5-fold gain in imaging signal by using the TSTA system over the one-step system. The TSTA approach will be a valuable and generalizable tool to amplify and noninvasively image reporter gene expression in living animals by using tissue-specific promoters. The approaches validated should have important implications for study of gene therapy vectors, cell trafficking, transgenic models, as well as studying development of eukaryotic organisms. PMID- 11734654 TI - A tumor host range selection procedure identifies p150(sal2) as a target of polyoma virus large T antigen. AB - Cancer cells may undergo loss or alterations in functions that certain viruses normally target to promote virus replication. Virus mutants that have lost the targeting function(s) should be able to grow in such cancer cells but not in normal cells. A "tumor host range" (t-hr) selection procedure has been devised and applied to polyoma virus based on this rationale. Studies of one t-hr mutant have led to the identification of the mSal2 gene product (p150(sal2)) as a binding partner of the large T antigen. mSal2 encodes a multizinc finger protein and putative transcription factor homologous to the Drosophila homeotic gene Spalt. The t-hr mutant encodes an altered large T protein that fails to interact with p150(sal2) and is defective in replication and tumor induction in newborn mice. PMID- 11734655 TI - Mechanisms underlying rapid experience-dependent plasticity in the human visual cortex. AB - Visual deprivation induces a rapid increase in visual cortex excitability that may result in better consolidation of spatial memory in animals and in lower visual recognition thresholds in humans. gamma-Aminobutyric acid (GABA)ergic, N methyl-d-aspartate (NMDA), and cholinergic receptors are thought to be involved in visual cortex plasticity in animal studies. Here, we used a pharmacological approach and found that lorazepam (which enhances GABA(A) receptor function by acting as a positive allosteric modulator), dextrometorphan (NMDA receptor antagonist), and scopolamine (muscarinic receptor antagonist) blocked rapid plastic changes associated with light deprivation. These findings suggest the involvement of GABA, NMDA, and cholinergic receptors in rapid experience dependent plasticity in the human visual cortex. PMID- 11734657 TI - Ricin A chain without its partner B chain is degraded after retrotranslocation from the endoplasmic reticulum to the cytosol in plant cells. AB - When expressed in tobacco cells, the catalytic subunit of the dimeric ribosome inactivating protein, ricin, is first inserted into the endoplasmic reticulum (ER) and then degraded in a manner that can be partially inhibited by the proteasome inhibitor clasto-lactacystin beta-lactone. Consistent with the implication of cytosolic proteasomes, degradation of ricin A chain is brefeldin A insensitive and the polypeptides that accumulate in the presence of the proteasome inhibitor are not processed in a vacuole-specific fashion. Rather, these stabilized polypeptides are in part deglycosylated by a peptide:N-glycanase like activity. Taken together, these results indicate that ricin A chain, albeit a structurally native protein, can behave as a substrate for ER to cytosol export, deglycosylation in the cytosol, and proteasomal degradation. Furthermore, retrotranslocation of this protein is not tightly coupled to proteasomal activity. These data are consistent with the hypothesis that ricin A chain can exploit the ER-associated protein degradation pathway to reach the cytosol. Although well characterized in mammalian and yeast cells, the operation of a similar pathway to the cytosol of plant cells has not previously been demonstrated. PMID- 11734656 TI - PTP and LTP at a hippocampal mossy fiber-interneuron synapse. AB - The mossy fiber-CA3 pyramidal neuron synapse is a main component of the hippocampal trisynaptic circuitry. Recent studies, however, suggested that inhibitory interneurons are the major targets of the mossy fiber system. To study the regulation of mossy fiber-interneuron excitation, we examined unitary and compound excitatory postsynaptic currents in dentate gyrus basket cells, evoked by paired recording between granule and basket cells or extracellular stimulation of mossy fiber collaterals. The application of an associative high-frequency stimulation paradigm induced posttetanic potentiation (PTP) followed by homosynaptic long-term potentiation (LTP). Analysis of numbers of failures, coefficient of variation, and paired-pulse modulation indicated that both PTP and LTP were expressed presynaptically. The Ca(2+) chelator 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) did not affect PTP or LTP at a concentration of 10 mM but attenuated LTP at a concentration of 30 mM. Both forskolin, an adenylyl cyclase activator, and phorbolester diacetate, a protein kinase C stimulator, lead to a long-lasting increase in excitatory postsynaptic current amplitude. H-89, a protein kinase A inhibitor, and bisindolylmaleimide, a protein kinase C antagonist, reduced PTP, whereas only bisindolylmaleimide reduced LTP. These results may suggest a differential contribution of protein kinase A and C pathways to mossy fiber-interneuron plasticity. Interneuron PTP and LTP may provide mechanisms to maintain the balance between synaptic excitation of interneurons and that of principal neurons in the dentate gyrus-CA3 network. PMID- 11734658 TI - Phospholipid signaling in plants: holding on to phospholipase D. AB - Plant cells contain various phospholipase-based signaling pathways. In fact, their repertoire of phospholipase D (PLD) molecules far outnumbers those of mammalian and yeast cells. Munnik and Musgrave take a broad look at PLD function in animal, yeast, and plant cells, and suggest that a PLD-based connection between membranes and microtubules is a biological property worth considering across species. PMID- 11734659 TI - The complex and intriguing lives of PIP2 with ion channels and transporters. AB - Phosphatidylinositol-4,5-bisphosphate (PIP(2)), the precursor of several signaling molecules in eukayotic cells, is itself also used by cells to signal to membrane-associated proteins. PIP(2) anchors numerous signaling molecules and cytoskeleton at the cell membrane, and the metabolism of PIP(2) is closely connected to membrane trafficking. Recently, ion transporters and channels have been discovered to be regulated by PIP(2). Systems reported to be activated by PIP(2) include (i) plasmalemmal calcium pumps (PMCA), (ii) cardiac sodium-calcium exchangers (NCX1), (iii) sodium-proton exchangers (NHE1-4), (iv) a sodium magnesium exchanger of unknown identity, (v) all inward rectifier potassium channels (KATP, IRK, GIRK, and ROMK channels), (vi) epithelial sodium channels (ENaC), and (vii) ryanodine-sensitive calcium release channels (RyR). Systems reported to be inhibited by PIP(2) include (i) cyclic nucleotide-gated channels of the rod (CNG), (ii) transient receptor potential-like (TRPL) Drosophila phototransduction channels, (iii) capsaicin-activated transient receptor potential (TRP) channels (VR1), and (iv) IP(3)-gated calcium release channels (IP3R). Systems that appear to be completely insensitive to PIP(2) include (i) voltage-gated sodium channels, (ii) most voltage-gated potassium channels, (iii) sodium-potassium pumps, (iv) several neurotransmitter transporters, and (v) cystic fibrosis transmembrane receptor (CFTR)-type chloride channels. Presumably, local changes of the concentration of PIP(2) in the plasma membrane represent cell signals to those mechanisms sensitive to PIP(2) changes. Unfortunately, our understanding of how local PIP(2) concentrations are regulated remains very limited. One important complexity is the probable existence of phospholipid microdomains, or lipid rafts. Such domains may serve to localize PIP(2) and thereby PIP(2) signaling, as well as to organize PIP(2) binding partners into signaling complexes. A related biological role of PIP(2) may be to control the activity of ion transporters and channels during biosynthesis or vesicle trafficking. Low PIP(2) concentrations in the secretory pathway would inactivate all of the systems that are stimulated by PIP(2). How, in detail, is PIP(2) used by cells to control ion channel and transporter activities? Further progress requires an improved understanding of lipid kinases and phosphatases, how they are regulated, where they are localized in cells, and with which ion channels and transporters they might localize. PMID- 11734660 TI - Alterations to haemostasis following cardiopulmonary bypass and the relationship of these changes to neurocognitive morbidity. AB - Cardiopulmonary bypass (CPB) is routinely utilized to provide circulatory support during cardiac surgical procedures. The morbidity of CPB has been significantly reduced since its introduction 50 years ago; however, cerebral injury remains a potentially serious consequence of otherwise successful surgery. The risk of stroke postoperatively is approximately 1-5%. Incidence rates for neurocognitive deficit, however, vary markedly depending on the detection method, although typically it is reported in at least 50% of patients. The aetiology of this cerebral injury remains open to debate, although evidence shows that ischaemia secondary to microembolism may be the principal factor. Emboli originate from bubbles of air, atheroemboli released on aortic manipulation and thromboemboli generated as a result of haemostatic activation. Significant generation of thrombin occurs during CPB resulting in fibrin formation, although the trigger of this activation is not fully understood. Rather than originating from contact activation as previously thought, the primary trigger may be via the activated factor VII/tissue factor pathway of coagulation, with an additional role of contact activation in amplification of coagulation as well as the fibrinolytic response to CPB. Haemostatic activation is inhibited with systemic heparin therapy. The relationship between haemostatic activation and emboli formation during CPB is not known. Interventions to reduce cerebral injury in the context of cardiac surgery depend, in large part, on the minimization of emboli. This review investigates cerebral injury after cardiac surgery and evidence showing that microembolism is the principal causative agent. Fibrin emboli are postulated to be an important source of cerebral embolism. The mechanism of haemostatic activation during CPB is therefore also discussed. PMID- 11734661 TI - Cost comparison of tinzaparin versus enoxaparin as deep venous thrombosis prophylaxis in spinal cord injury: preliminary data. AB - Thromboembolic events are major causes of morbidity and mortality in patients with spinal cord injuries. Low molecular weight heparins are recommended as prophylaxis against such events. The purpose of the current study was to perform a cost analysis of tinzaparin versus enoxaparin using published efficacy and safety data as deep vein thrombosis and pulmonary embolism prophylaxis in this population. All published English language articles evaluating either tinzaparin or enoxaparin as pharmacoprophylaxis in spinal cord-injured patients were identified. Data from these studies were subjected to cost-effectiveness analyses followed by sensitivity analyses to determine which agent is the most cost effective in these patients. Results demonstrated that tinzaparin 3500 U daily and enoxaparin 30 mg every 12 h are both cost-effective agents for thromboembolism prophylaxis in patients with spinal cord injuries. PMID- 11734662 TI - Disintegration and reorganization of fibrin networks during tissue-type plasminogen activator-induced clot lysis. AB - In this study, we investigated tissue-type plasminogen activator (tPA)-induced lysis of glutamic acid (glu)-plasminogen-containing or lysine (lys)-plasminogen containing thrombin-induced fibrin clots. We measured clot development and plasmin-mediated clot disintegration by thromboelastography, and used scanning electron microscopy (SEM) to document the structural changes taking place during clot formation and lysis. These events occurred in three overlapping stages, which were initiated by the addition of thrombin, resulting first in fibrin polymerization and clot network organization (Stage I). Autolytic plasmin cleavage of glu-plasminogen at lys-77 generates lys-plasminogen, exposing lysine binding sites in its kringle domains. The presence of lys-plasminogen within the thrombin-induced fibrin clot enhanced network reorganization to form thicker fibers as well as globular complexes containing fibrin and lys-plasminogen having a greater level of turbidity and a higher elastic modulus (G) than occurred with thrombin alone. Lys-plasminogen or glu-plasminogen that had been incorporated into the fibrin clot was activated to plasmin by tPA admixed with the thrombin, and led directly to clot disintegration (Stage II) concomitant with fibrin network reorganization. The onset of Stage III (clot dissolution) was signaled by a sustained secondary rise in turbidity that was due to the combined effects of lys-plasminogen presence or its conversion from glu-plasminogen, plus clot network reorganization. SEM images documented dynamic structural changes in the lysing fibrin network and showed that the secondary turbidity rise was due to extensive reorganization of severed fibrils and fibers to form wide, occasionally branched fibers. These degraded structures contributed little, if anything, to the structural integrity of the residual clot, and eventually collapsed completely during the course of progressive clot dissolution. These results provide new perspectives on the major structural events that occur in the fibrin clot matrix during fibrinolysis. PMID- 11734663 TI - Prevalence of factor V Leiden and prothrombin 20210 A variant in Bulgarian patients with pulmonary thromboembolism and deep venous thrombosis. AB - Factor V Leiden mutation and prothrombin variant 20210 A are well-known risk factors for venous thrombosis (DVT). Recent papers have reported a lower prevalence of factor V Leiden in patients with pulmonary thromboembolism (PTE) than in patients with deep venous thrombosis. The aim of the present study was to compare the prevalence of factor V Leiden and the prothrombin 20210 G <-- A mutation in patients with DVT and in patients with PTE. We studied 128 consecutive patients (45 with DVT, 40 with PTE, and 43 with DVT and PTE) for factor V Leiden and prothrombin 20210 A. One hundred healthy persons matched by age and sex were used as controls. Factor V Leiden was present in five of the patients with PTE [12.5%; 95% confidence interval (CI), 1.5-23.5%; not significant], 15 of the patients with DVT (33.3%; 95% CI, 9.6-38.7%; P < 0.001), and 12 of the patients with DVT and PTE (27.9%; 95% CI, 4.8-33%; P = 0.001). Results for the prothrombin 20210 A mutation were as follows: four of 40 patients with PTE (10%; 95% CI, 0-13.3%; P = 0.46), nine of 45 (20%) of the patients with DVT (95% CI, 0.5-25.5%; P < 0.05) and eight of 43 with DVT and PTE were heterozygous (18.6%; 95% CI, 0-23.9%; P = 0.02). In conclusion, there is a significantly higher frequency of factor V Leiden among patients with DVT than in patients with PTE. However, there is no significant difference of factor V Leiden or 20210 A prothrombin mutation in patients with DVT than in patients with combined DVT/PTE, therefore patients with DVT, carriers of the mutations, do not appear to be at lower risk for pulmonary embolism. PMID- 11734664 TI - Mean transit times and the sites of synthesis and catabolism of tissue plasminogen activator and plasminogen activator inhibitor type 1 in young subjects. AB - Using an invasive technique, we studied the mean transit time, the net quantitative turnover rate, and the sites of synthesis and catabolism of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) in healthy young volunteers in the fasting, steady state. Blood was sampled simultaneously from a large hepatic vein, an artery and the inferior caval vein, while measuring the splanchnic plasma flow rate and the plasma volume. We found that the catabolism of active t-PA and t-PA antigen took place in the splanchnic circulation with net rates of 7.2 and 6.3 pmol/min, respectively. The extraction fraction and the mean transit time in the splanchnic circulation were, respectively, 0.63 and 5.6 min for active t-PA and 0.17 and 21 min for t-PA antigen. Active PAI-1 was synthesized in the splanchnic circulation at a rate of 890 IU/min and had a mean transit time of about 9.8 min. No net extraction of PAI 1 antigen took place in the splanchnic circulation. In conclusion, we demonstrated that active t-PA and t-PA antigen are catabolized and active PAI-1 produced in the splanchnic circulation in young healthy subjects during steady state. Furthermore, our data show that active t-PA was also eliminated outside the splanchnic region with a catabolism rate of about 8.4 pmol/min. No net complex formation could be demonstrated in the peripheral circulation. We therefore suggest that active t-PA is eliminated by a re-uptake in the endothelium in the peripheral vessels or in the lung circulation. PMID- 11734665 TI - Haemostasis in ischaemic stroke and vascular dementia. AB - Abnormalities of coagulation and fibrinolysis may play an important role in the pathogenesis of ischaemic stroke and vascular dementia. We aimed to determine whether haemostatic function is altered in acute recent-onset or chronic ischaemic cerebrovascular disease. We studied consecutive patients with ischaemic stroke (n = 74) and vascular dementia (n = 42) compared with healthy controls (n = 40) in a case-control study. The ischaemic stroke group was assessed twice, 3 10 days after the acute stroke and at 1-3 months. Fibrinogen, fibrin D-dimer (marker of fibrin turnover) and von Willebrand factor (vWF) (marker of endothelial disturbance) were elevated acutely (P < 0.0001) and in the convalescent phase after ischaemic stroke (P < 0.0001, P < 0.0001, and P < 0.01 respectively, compared with controls). Similar results were seen in the vascular dementia group. Stepwise multivariate regression analyses showed that cerebrovascular disease correlated independently with fibrinogen (P < 0.001) and fibrin D-dimer levels (P < 0.001), while vWF correlated independently with electrocardiograph evidence of ischaemic heart disease (P = 0.004). Changes between acute and convalescent phases in ischaemic stroke were slightly inconsistent. However, in the acute stage there were tendencies for fibrinogen, D dimer and vWF to be increased, and factor VIII was significantly higher. Abnormalities of haemostasis, including increased fibrin turnover and endothelial disturbance, are found in both acute and chronic cerebral ischaemia. Many of these patients have co-existent ischaemic heart disease and this may contribute to some of these changes. Acute ischaemic stroke is associated with transient changes in haemostatic factors; however, most abnormalities persist into the convalescent phase, and are also demonstrable in subjects with vascular dementia. PMID- 11734666 TI - Comparative prevalence of antiphospholipid antibodies and thrombophilic genotypes in consecutive patients with venous thrombosis. AB - The prevalence of antiphospholipid antibodies (aPL) and thrombophilic genotypes was compared and their reciprocal interactions assessed in consecutive patients with venous thrombosis (n = 101; 58 male, 43 female; mean age, 56 +/- 16 years) and in blood donors (n = 121; 55 male, 46 female; mean age, 43 +/- 12 years). Multiple aPL were detected by enzyme-linked immunosorbent assay (ELISA) and factor V Leiden (FVL), methylenetetrahydrofolate reductase (MTHFR) C677 <-- T and prothrombin (PT) G20210 <-- A by polymerase chain reaction. A pro-thrombotic state, including aPL, was found in 81% (82/101) of patients and 23% (29/121) of controls (P < 0.0001, odds ratio = 13.69, 95% confidence interval = 7.14-26.25). The prevalence of total aPL-positives was lower than MTHRF+/+ (homozygous state), FVL and PT G20210 <-- A (P < 0.0001) in the control group, but in line with the prevalence of total thrombophilic genotypes in the patient group: MTHFR+/+, 29%; FVL, 19%; PT, 14%; aPL, 20%. Having variously combined aPL and thrombophilic genotypes, only aPL remained as the most common pro-thrombotic factor in the patient group (P = 0.003). In patients without circumstantial factors for thrombosis, carriers of MTHFR+/+ + aPL showed a lower age at event than carriers of MTHFR+/+ alone or aPL alone (38 +/- 24 versus 65 +/- 10 years, P < 0.05 and 38 +/- 24 versus 63 +/- 16 years, P < 0.05, respectively). In the same patients, mean plasma homocysteine measured by an ELISA method was significantly higher in the MTHFR+/+ + aPL group than in the MTHFR+/+ alone or aPL alone groups (P = 0.01). Antiphospholipid antibodies are as common as thrombophilic genotypes in patients with venous thrombosis, and the interaction of aPL with MTHFR+/+ may influence age at first event via elevated plasma homocysteine. PMID- 11734667 TI - Fibrinogen increases lipopolysaccharide-induced tumor necrosis factor-alpha and interleukin-8 release, and enhances tissue factor activity in monocytes in a modified whole blood system. AB - The objective of this study was to investigate the effect of fibrinogen on lipopolysaccharide (LPS)-stimulated blood cells. To this end, a minimum essential blood system was established, reconstituted from washed blood cells and 20% (fibrinogen-free) lepirudin anticoagulated serum in RPMI-1640. Concurrent addition to the system of 1.0-4.0 mg/ml fibrinogen increased LPS-induced tissue factor (TF) activity in the monocytes in a dose-dependent manner. This enhancing effect was, by and large, independent of the LPS concentration (0.5-5.0 ng/ml). Even at the lowest concentration of fibrinogen (1.0 mg/ml), the enhancing effect was quite significant (46-80%) at almost every concentration of LPS tested. Furthermore, LPS-induced release of the two proinflammatory products tumor necrosis factor-alpha and interleukin-8 were also enhanced by added fibrinogen. In conclusion, fibrinogen is capable of enhancing the emergence of certain proinflammatory molecules as well as the procoagulant factor TF, effects that may very well in part be accountable for fibrinogen-related risk of ischemic heart disease and stroke. PMID- 11734668 TI - Hypercoagulation in surgical postmenopausal women having hormone replacement with overdose estradiol. AB - A benefit of estradiol replacement preventing coronary heart disease (CHD) after menopause has been suggested by clinical investigations. In the department of gynecology at our hospital, we met by chance eight senior women who were hysterectomized due to different benign gynecological disorders. Acting on their own, they took a daily dose of estradiol valerate as high as 8-50 mg for 3 years, in comparison with 1-2 mg used in routine hormonal replacement therapy. We were interested to assess whether the overdose estrogen disturbs the hemostatic function, thus losing the favorable effect on CHD occurrence. Plasma levels of some procoagulants/anticoagulants were assayed in the eight women with replacement therapy and the results were compared with those of seven age-matched senior women and of 14 young healthy women with normal menstrual cycles. Using a new laboratory method recently developed by us, the overall hemostatic potential, shown as a single parameter (Abs-sum), was also determined. Results showed that high-dose estradiol activated coagulation and depressed fibrinolysis, leading to the net effect of elevated overall hemostatic potential. Thus, the overdose estradiol replacement may not be beneficial in preventing CHD after menopause, or it may even increase the risk due to the hypercoagulable state induced therefrom. PMID- 11734669 TI - Investigation of genotype-dependent differences in factor V activity as well as response to activated protein C by application of different methods. AB - Coagulation factor V has been at the centre of investigation for several years. In addition to factor V Leiden, various other polymorphisms are becoming the object of interest. Different results have been published about the association of the HR2 haplotype with decreased factor V levels and with reduced response to activated protein C (APC). Due to the central position of factor V in the clotting process, its activity can be determined in both thromboplastin-based and activated partial thromboplastin time (aPTT)-based assays. A multitude of assays are known for the determination of APC response. The aim of our study was to investigate whether different methods disclose genotype-dependent differences in factor V activity as well as APC response. Three wild-type carriers, three carriers homozygous for the R2 allele (4070G), and three carriers homozygous for the G allele (2391G, 2663G, 2684G, 2863G) were investigated. For each individual plasma sample, the factor V activity was determined using 12 different reagent combinations of three different thromboplastins, three different aPTT reagents, and two different factor V deficient plasma sources. The determination of factor V activity in the thromboplastin system revealed differences between the genotypes. These differences were independent of the thromboplastin reagent and the factor V-deficient plasma. The aPTT system exhibited a dependency on the aPTT reagent and the factor V-deficient plasma. Analysis of APC response disclosed genomic differences in specific test systems only. One type of assay could be more appropriate than other types in dependence of the position of genomic variations. Therefore, the applied assay is an important influential factor in investigations of functional consequences of genomic variations. PMID- 11734670 TI - The reduced sensitivity of the ProC Global test in protein S deficient subjects reflects a reduction in the associated thrombotic risk. AB - To investigate simultaneously a defect affecting the protein C/protein S (PC/PS) anticoagulant pathway is possible thanks to a methodological approach (ProC(R) Global; Dade Behring) based on the activation of endogenous plasma PC by a snake venom extract. Factor V (FV) Leiden, the most frequent cause of hereditary thrombosis, is well detected by the test with sensitivity of 100% irrespective of the presence/absence of thrombosis in the subjects investigated. The test is also suited to detect PC or PS defect, but in this case the in vitro impairment of the PC/PS pathway is less pronounced particularly for PS defects (sensitivity for PC and PS defect, 85-100 and 30-90%, respectively). In this study, we hypothesized that the lower sensitivity described for PS defect, compared with those of PC and FV Leiden defects, could also be related to the clinical condition of the subject investigated (symptomatic/asymptomatic) rather than solely to the PS plasma activity/level. Therefore, we analyzed 126 subjects with single congenital defects in the PC/PS pathway: 46 subjects with PS deficiency (26 thrombotic cases and 20 asymptomatic relatives), 40 subjects with PC deficiency (25 thrombotic cases and 15 asymptomatic relatives), and 40 heterozygous FV Leiden subjects (25 thrombotic cases and 15 asymptomatic relatives). By a cut-off of normalized Agkistrodon contortix snake venom ratio of 0.84, the sensitivity in the whole group of cases (sensitivity a) was 76.1, 95.0 and 100%, respectively, for PS, PC and FV Leiden defects. The test failed to detect 11 (23.9%) among the 46 PS deficient subjects, and all these cases except two belonged to the asymptomatic subgroup (9/20; 45%). Excluding the 20 asymptomatic relatives, the new sensitivity (sensitivity b) for the PS defect was 92.3%. The comparison of the sensitivity in the symptomatic PS cases and in the asymptomatic ones was significantly different (P = 0.010). Among the 40 PC-deficient subjects, only two (5.0%) were not detected by the test and they belonged indifferently to the two subgroups. Finally, none of the 40 FV Leiden heterozygotes were misdiagnosed by the test. These results suggest that in symptomatic PS-deficient cases the test could reflect a post-thrombotic effect and/or reveal potential unidentified prothrombotic influences assessing a prothrombotic risk condition. PMID- 11734671 TI - Impact of plasma homocysteine and prothrombin G20210 A on primary antiphospholipid syndrome. AB - The prevalence of prothrombin (PT) G20210A and methylenetetrahydrofolate reductase (MTHFR) C677 <-- T was assessed in 40 patients with primary antiphospholipid syndrome (APS) (14 male, 26 female; mean age, 37 +/- 14 years) and in 27 persistent carriers of antiphospholipid antibodies (aPL) (five male, 22 female; mean age, 40 +/- 16 years) without underlying diseases. Non-APS thrombotic patients (n = 100; 47 female, 53 male; mean age, 40 +/- 10 years) and healthy subjects (n = 100; 46 female, 54 male; mean age, 56 +/- 16 years) served as control groups. Plasma homocysteine (HC) (enzyme-linked immunosorbent assay) was measured in all aPL patients and in 51 subjects from the healthy control group (mean age, 38 +/- 16 years). Heterozygous prothrombin PT G20210A was more frequent in the thrombotic group without APS (18%) than in the control (4%), APS (12%) or aPL (11%) groups, whereas homozygous MTHFR C677 <-- T was equally distributed. After genotype sub-grouping, plasma HC was higher in APS patients with homozygous MTHFR C677 <-- T compared with non-homozygous APS patients (22 +/ 5.4 versus 11 +/- 1.3 micromol/l; P < 0.01) and with homozygous MTHFR C677 <-- T controls (22 +/- 5.4 versus 15 +/- 2.0 micromol/l). In the APS group, mean age at first event was lower in homozygous MTHFR C677 <-- T patients than in non homozygous patients (26 +/- 7.5 versus 36 +/- 13 years; P = 0.008). In the same group, homozygous MTHFR C677 <-- T patients suffered an increased average number of events per person than non-homozygous patients (1.9 versus 1.3; P = 0.04). Heterozygous PT G20210A contributes little to the thrombotic tendency of primary APS whereas plasma HC may influence age at first event and number of events. Measurement of plasma HC in aPL subjects may identify patients at increased thrombotic risk requiring HC lowering. PMID- 11734672 TI - Alterations in the extrinsic pathway in hypertriglyceridemia do not cause a 'procoagulant state': effects of bezafibrate therapy. AB - Hypertriglyceridemia (HTG) is an independent risk factor for cardiovascular disease (CVD). Hemostatic variables [factor VII antigen (FVIIag), factor VII coagulant activity (FVIIc), activated factor VII (FVIIa), free and endothelial associated (EC) tissue factor pathway inhibitor (TFPI) antigen, pre- and post heparin total TFPI activity, EC-TFPI activity, prothrombin fragment 1 + 2 (F1 + 2), fibrinogen and D-dimer] were compared between 18 HTG patients and 20 controls to investigate whether HTG is associated with alterations in the extrinsic pathway and whether such alterations create a procoagulant state, as expressed by F1 + 2 and D-dimer levels. In addition, the effects of bezafibrate therapy (6 weeks, 400 mg/day) on these variables were studied in 18 HTG patients in a double blind, placebo-controlled, cross-over study. FVIIag, FVIIc, free TFPI and fibrinogen were significantly higher in HTG patients (by 44, 30, 45 and 31%, respectively; all P < 0.02), while FVIIa, EC-TFPIag and activity, total TFPI activities, F1 + 2 and D-dimer levels were similar in patients and controls. Bezafibrate reduced serum TG and fibrinogen levels (by 62 and 20%, respectively; both P < 0.001), whereas the other hemostatic variables were unaffected. In conclusion, the observed alterations in the extrinsic pathway in HTG are not associated with a procoagulant state. In contrast, the presence of elevated fibrinogen levels in HTG might enhance the risk for CVD. Bezafibrate therapy improved the adverse lipid profile and decreased fibrinogen levels in HTG patients. PMID- 11734673 TI - Risk of venous thromboembolism in carriers of factor V Leiden with a concomitant inherited thrombophilic defect: a retrospective analysis. AB - Factor V Leiden is the most common genetic defect associated with venous thromboembolism. Its clinical expression is limited and shows a wide intrafamilial and interfamilial variation, which might be explained by the influence of other genetic risk factors. We retrospectively studied 226 patients with factor V Leiden and documented venous thromboembolism (probands) and 400 first-degree carrier relatives to assess the contribution of concomitant genetic risk factors to the occurrence of venous thromboembolism. The prothrombin G20210A mutation was found in 8.3%, homozygosity of factor V Leiden in 7.2%, and inherited deficiencies of antithrombin, protein C or protein S in 4.7% of symptomatic carriers (probands and relatives), as compared with 6.0, 3.4 and 0.9% of asymptomatic carriers, respectively. The total follow-up time in relatives was 11 049 years. Prevalences of venous thromboembolism were 10.8% in single heterozygous factor V Leiden carrier relatives, 16.0% in double-heterozygotes for factor V Leiden and the prothrombin mutation, 36.8% in homozygotes for factor V Leiden, and 40.0% in double-heterozygotes for factor V Leiden and an inherited deficiency of protein C or protein S. Annual incidences in these groups were 0.39, 0.57, 1.41, and 4.76%, respectively. Multivariate analysis showed a small, non-significant additional effect of the prothrombin mutation on the risk of venous thromboembolism in heterozygotes for factor V Leiden [adjusted hazard ratio, 1.3; 95% confidence interval (CI), 0.5-3.8]. This effect was more pronounced for homozygosity of factor V Leiden (adjusted hazard ratio, 3.9; 95% CI, 1.7-9.0) and inherited protein C or protein S deficiencies (adjusted hazard ratio, 17.5; 95% CI, 3.8-81.2). Our data provide evidence of clustering of the evaluated genetic thrombophilic defects in symptomatic factor V Leiden carriers and support the assumption that the clinical expression of factor V Leiden depends on clustering in a part of carriers. PMID- 11734674 TI - Lifestyle factors and coagulation activation markers: the Caerphilly Study. AB - Coagulation activation markers are being investigated as risk factors for cardiovascular disease; we examined the contribution of several lifestyle factors to variation in plasma levels of several activation markers measured in a population-based study of 1947 men. Smoking, alcohol, body mass index (BMI), leisure and work activity, social class, and use of prescribed medicines were each examined in turn. Specific assays of fibrin D-dimer and von Willebrand factor (vWF) activity showed similar relationships to lifestyle variables as we observed previously for less specific assays of D-dimer and vWF antigen. D-dimer levels increased with age, in smokers and in men taking prescribed medication, and were negatively associated with leisure time activity. vWF activity increased with age and showed a U-shaped distribution with BMI. Factors VIIc and VIIIc and thrombin-antithrombin complexes were associated with BMI, factor VIIIc and prothrombin fragments 1 + 2 (F1 + 2) were associated with age, activated partial thromboplastin time (aPTT) and F1 + 2 were associated with smoking, and aPTT showed a small negative association with alcohol consumption. We conclude that lifestyle modification has the possibility of favourably influencing several of these risk markers. In particular, cigarette smoking has a possibly reversible effect on coagulation activation (measured by F1 + 2 and D-dimer). PMID- 11734675 TI - Association between decreased pulmonary endothelial cell thrombomodulin and local fibrin deposition in pneumonia. AB - Thrombomodulin (TM) plays an important role in anticoagulation by forming a complex with thrombin, which subsequently activates protein C. TM is inactivated and downregulated by inflammatory cell mediators. This study examined whether bronchopneumonia is associated with changes in TM immunoreactivity, and whether a decrease in TM is accompanied by evidence of hypercoagulability, i.e. local deposition of fibrin. Double antibody staining for TM and fibrin was performed on lung tissue sections from patients who had died of pneumonia and from patients who had died rapidly, secondary to trauma. Inflammatory changes were assessed histologically and immunohistochemically using antibodies against interleukin 1alpha, tumor necrosis factor-alpha, and myeloperoxidase. Areas with bronchopneumonia exhibited markedly decreased endothelial TM staining of alveolar walls and small vessels. These changes were associated with prominent fibrin immunoreactivity. Some areas exhibited mild to moderate inflammation with little fibrin deposition and variable amounts of TM in adjacent vessels. This study is the first to relate changes of TM immunoreactivity levels to fibrin deposition in a human disease process. These data may have implications for pulmonary pathophysiology in patients with bronchopneumonia. PMID- 11734677 TI - Evidence-based neuro-ophthalmology: advances in treatment. AB - This review presents highlights and updates from of the most significant clinical trials in neuro-ophthalmology to date, the Optic Neuritis Treatment Trial, the Controlled High-Risk Avonex Multiple Sclerosis Prevention Study, and the Ischemic Optic Neuropathy Decompression Trial. The quality of evidence for treatment efficacy from these trials and other recent investigations of giant cell arteritis and idiopathic intracranial hypertension is classified herein according to published criteria based on sample size and study design. PMID- 11734678 TI - Giant cell arteritis: diagnosis and management. AB - Visual loss caused by giant cell arteritis is a medical emergency that requires prompt recognition and treatment with systemic corticosteroids. A delay in diagnosis can lead to devastating ophthalmic and systemic complications. Recent advances in genetic and immunocytochemical research techniques have led to greater understanding of the underlying pathomechanisms of giant cell arteritis. Giant cell arteritis is a systemic condition with a strong predilection for the ocular vasculature. Visual symptoms are often the presenting manifestation of the disease, placing the ophthalmologist in a critical position for early diagnosis and treatment. Maintenance of a high clinical suspicion in the appropriate clinical setting is important in establishing an early diagnosis. Because of the complex nature of the disease process, many cases may be a therapeutic challenge requiring prolonged immunosuppression. PMID- 11734679 TI - Management of optic neuritis and multiple sclerosis. AB - Optic neuritis and multiple sclerosis (MS) are common causes of visual and neurologic dysfunction in young adults. Advances in magnetic resonance imaging, molecular genetics, and neuroimmunology have increased our understanding of the pathophysiology underlying both disorders. Corticosteroids remain the mainstay of treatment of optic neuritis, but alternate dosages and routes of administration are undergoing investigation. The available therapies for MS have expanded, and there is evidence that early intervention is beneficial. Treatments for MS show sustained efficacy, but are not curative, and adjunctive treatments may prove valuable in patients with progressive visual and neurologic disability. PMID- 11734680 TI - Strabismus surgery in the treatment of paralytic strabismus. AB - Strabismus surgery may restore limited binocular function and improve cosmesis in patients with paralytic strabismus. Evaluation of the amount of residual function of the affected extraocular muscles is essential to determine which surgical procedure will yield the best results. Standard muscle resection techniques are effective for patients who have paretic extraocular muscles with residual function. Muscle transposition procedures are indicated for patients with total paralysis of an extraocular muscle. Autogenous and alloplastic materials may be required to fix the eye in primary position in patients with total paralysis of multiple extraocular muscles. This article discusses the recent literature regarding the evaluation and management of patients with paralytic strabismus. Current concepts regarding extraocular muscle transposition and the use of autogenous materials are emphasized. PMID- 11734681 TI - Use of botulinum toxin in neuro-ophthalmology. AB - Botulinum toxin produces a temporary chemical denervation, which initially was used to reduce the diplopia associated with paralytic strabismus. Subsequently, the drug was found to be effective in the management of dystonias, including hemifacial spasm and blepharospasm. More recently, there is evidence that botulinum toxin may lessen the pain associated with some movement disorders, tension headache, and migraine. The effect of botulinum toxin in temporary, but the therapeutic benefit usually is maintained even after repeated injections. The form of botulinum toxin used in clinical practice is the A serotype. Other serotypes may have a therapeutic role in the future. The mode of action and the clinical experience of botulinum toxin in neuro-ophthalmology is discussed. PMID- 11734682 TI - Functional magnetic resonance imaging of the visual system. AB - Functional magnetic resonance imaging (fMRI), which is a technique useful for non invasive mapping of brain function, is well suited for studying the visual system. This review highlights current clinical applications and research studies involving patients with visual deficits. Relevant reports regarding the investigation of the brain's role in visual processing and some newer fMRI techniques are also reviewed. Functional magnetic resonance imaging has been used for presurgical mapping of visual cortex in patients with brain lesions and for studying patients with amblyopia, optic neuritis, and residual vision in homonymous hemianopia. Retinotopic borders, motion processing, and visual attention have been the topics of several fMRI studies. These reports suggest that fMRI can be useful in clinical and research studies in patients with visual deficits. PMID- 11734683 TI - Ocular syphilis. AB - Although the annual incidence of primary and secondary syphilis has dropped to the lowest rate recorded, syphilis remains an important cause of ocular disease. Uveitis is the most common ocular manifestation of syphilis in both HIV-positive and HIV-negative patients, and the diagnosis should prompt an analysis of the cerebrospinal fluid to exclude associated neurosyphilis. Newer modalities such as enzyme immunoassays and genomic amplification using the polymerase chain reaction may prove to be useful techniques to detect Treponema pallidum in intraocular specimens. The preferred treatment for all stages of syphilis remains parenteral penicillin G, although the preparation, dose, route of administration, and duration of therapy are dictated by the stage of disease and various host factors. All patients diagnosed with ocular syphilis should be tested for HIV, because the presence of a primary genital chancre increases the risk of acquiring or transmitting HIV, and because risk factors for the two diseases are similar. PMID- 11734684 TI - Presumed ocular histoplasmosis syndrome: update on epidemiology, pathogenesis, and photodynamic, antiangiogenic, and surgical therapies. AB - Presumed ocular histoplasmosis involves the classic triad of discrete atrophic choroidal scars in the macula or midperiphery known as histo spots, peripapillary atrophy, and choroidal neovascularization, which leads to severe loss of central vision. The histo spots from which the choroidal neovascularization develop do not show active inflammation but do represent focal defects in Bruch membrane that could facilitate development of choroidal neovascularization. The macular photocoagulation studies unequivocally show the benefit of photocoagulation compared with observation in reducing the risk of vision loss in patients with presumed ocular histoplasmosis, well defined extrafoveal or juxtafoveal choroidal neovascularization, and choroidal neovascularization in the peripapillary area. However, laser treatment itself causes an absolute scotoma correlating with the site of the laser photocoagulation scar, and subfoveal choroidal neovascularization is not amenable to laser photocoagulation because this would cause a blinding central scotoma. Consequently, other treatments have been sought. The Verteporfin in Ocular Histoplasmosis study evaluated photodynamic therapy for subfoveal choroidal neovascularization caused by presumed ocular histoplasmosis and demonstrated stabilization of the choroidal neovascularization and visual acuity benefit. In addition to photodynamic therapy, antiangiogenic compounds are being developed for choroidal neovascularization caused by age related macular degeneration, and these agents will likely be of benefit in presumed ocular histoplasmosis associated choroidal neovascularization. Finally, submacular surgery for the removal of subfoveal choroidal neovascularization has promising results. The results of these research efforts will produce more effective therapeutic approaches in the future. PMID- 11734685 TI - Nematode infections of the eye: toxocariasis and diffuse unilateral subacute neuroretinitis. AB - In many parts of the world, parasitic infections of the eye are a major cause of blindness. The parasites Toxocara canis, Onchocerca volvulus, Taenia solium, Ancylostoma caninum, and Cysticercus celulosae all have been responsible for blinding ocular infections. The nematodes T. canis and Toxocara cati are parasitic roundworms that infect dogs (toxocarosis), other canidae, and cats. Ocular toxocariasis is an uncommon worldwide infection caused by the nematode larvae of T. canis, commonly found in dogs. Human transmission is usually via geophagia, the ingestion of food contaminated with Toxocara eggs, or contact with infected puppies, often resulting in devastating ocular or systemic effects. Ocular toxocariasis is typically a monocular disease of young children, and its clinical findings include posterior and peripheral retinochoroiditis, optic papillitis, and endophthalmitis. The inflammatory response created by ocular involvement may result in epiretinal membrane formation, traction retinal detachment, and combined traction-rhegmatogenous retinal detachment. Diffuse unilateral subacute neuroretinitis is another ocular parasitic infection that usually results in severe visual loss. Evidence suggests that diffuse unilateral subacute neuroretinitis is caused by a solitary unidentified nematode of two different sizes, but to date, only a small number of nematodes have been recovered from eyes affected with the infection. Diffuse unilateral subacute neuroretinitis occasionally can affect the fellow eye. PMID- 11734686 TI - Ocular complications of pregnancy. AB - Pregnancy is often associated with ocular changes, most often transient in nature, though occasionally permanent. It can be associated with development of new conditions, or can exacerbate pre-existing conditions. The ocular effects of pregnancy may be divided into physiologic changes, pathologic conditions or modifications of pre-existing conditions. Pathologic conditions include entities such as pre-eclampsia and eclampsia, along with conditions that are seen with increased frequency during pregnancy such as central serous retinopathy. The most significant modified pre-existing condition is diabetes mellitus. The various effects of pregnancy on the eye will be reviewed in this article. PMID- 11734687 TI - Endogenous endophthalmitis. AB - Endogenous endophthalmitis still presents a diagnostic and therapeutic challenge even with improvements in therapeutic modalities. The main prognostic factor is the virulence of the causative organism: once the organism enters the eye, it rapidly destroys ocular tissues. With the increasing ability to prolong the life of very ill patients, incidence of endogenous endophthalmitis is expected to increase. Ophthalmologists must be ever more vigilant, because prompt treatment offers the best chance of saving the eye. PMID- 11734688 TI - Systemic disorders associated with episcleritis and scleritis. AB - Inflammation of the episclera and sclera still represents a diagnostic challenge. Many of these presentations are idiopathic and are managed empirically with the use of anti-inflammatory therapy, usually with a satisfactory response. However, some cases will have a more aggressive nature or will show resistance to conventional therapy, and those potentially are connected to an infectious or underlying disease. In such cases, there is an increased risk of visual loss and severe systemic complications requiring intensive investigations and aggressive therapy. The diseases more commonly associated with these conditions, especially with scleritis, are the connective tissue disorders and systemic vasculitis, but systemic infections and malignancies also must be excluded. Management depends on the specific diagnosis, and every year a new list of potential associations or atypical presentations is reported. For a successful outcome, it is important that these presentations are recognized. PMID- 11734690 TI - Surgery for urogenital rhabdomyosarcoma. AB - The treatment and prognosis of rhabdomyosarcoma of the urogenital tract have changed dramatically over the past few decades. Until the 1970s radical cystectomy and pelvic exenteration were the only therapeutic options in rhabdomyosarcoma of the bladder/prostate and of the vagina/uterus. Since then, multiagent chemotherapy has achieved remarkable cure rates. Consequently, organ saving surgical techniques were advocated, with the goal to improve patients' quality of life. The available long-term survival data with these techniques demonstrate excellent survival in rhabdomyosarcoma of the vagina/uterus, with a high percentage of organ preservation. On the contrary, increased rates of local recurrence and death from disease in patients with rhabdomyosarcoma of the bladder/prostate who undergo primary chemotherapy with or without tumour excision, and complications following combined radiotherapy critically question the long-term benefit of these organ-sparing surgical techniques in this group. PMID- 11734691 TI - Wilms' tumor (nephroblastoma). AB - Wilms' tumor (nephroblastoma) is a subject which continues to challenge clinicians in their attempts to achieve the best survival of their patients, while minimizing morbidity. Overall survival of over 85% of all children can now be achieved using combination therapy with chemotherapy, surgery, and in some cases radiotherapy. Recent reviews of two multi-institutional, multinational trials (the International Society of Pediatric Oncology and the National Wilms' Tumor Study Group) are discussed in this article, in which their current approaches to therapy are presented. In addition to these clinical advances, the genetic and molecular features of Wilms' tumor continue to provide insight into the biology of nephroblastoma and into neoplasia in general. A recent review of these aspects of the field is provided. PMID- 11734692 TI - Renal function in congenital anomalies of the kidney and urinary tract. AB - Congenital anomalies of the kidneys and urinary tract are a major cause of chronic and end-stage renal failure in children. The molecular mechanisms having been elaborated, there is now growing evidence that kidney function is to a large extent determined genetically at an early stage. Assessment of kidney function is an important tool in clinical medicine and is feasible in utero. Postnatally, determination of absolute glomerular filtration rate and also of split and excretory renal function play an important role in the determination of treatment and prognosis. This is supplemented by other biochemical, molecular and interventional prognostic factors, which are of help in preservation of kidney survival by minimizing modulating factors. If chronic or terminal renal failure ensues in childhood or even in early infancy, however, improved medical care has led to encouraging results, ultimately influencing the motivation in the care of children with congenital anomalies of the kidney and urinary tract. PMID- 11734693 TI - Neurogenic and non-neurogenic bladder dysfunction. AB - Children with a neurological defect have a clear cause for their bladder dysfunction; however, in neurologically normal children the cause of their incontinence is usually unclear. When no anatomical abnormalities seem to be present a functional problem is generally the cause. This type of incontinence is referred to as 'functional incontinence'. The different forms of bladder and sphincter dysfunction will be discussed and treatment modalities described. As the treatment modalities in children with neuropathic bladders focus on medical and especially surgical options, special attention is paid to new developments in surgical treatment. For those with functional incontinence treatment options are more variable and the new developments are described. PMID- 11734694 TI - Surgery to the external genitalia. AB - Surgery to the external genitalia is one of the most challenging chapters of reconstructive urology, due to the need to correct complex problems such as hypospadias or epispadias (isolated or associated to exstrophies). Although this specialty has shown a continuous technical advancement there is no consensus as to the most efficient surgical procedure or to the best strategy to treat these pathologies. The papers reviewed here reflect these statements and reinforce the idea that surgeons' good sense, allied with their experience in the area, are the factors that define the choice of the ideal surgical option for each case. PMID- 11734695 TI - New surgical techniques in pediatric urology. AB - More sophisticated endoscopic instruments, combined with a better understanding of bladder and urethral pathology, have significantly improved the therapeutic approaches for both posterior urethral valves and ureteroceles. New generation lithotripters have allowed for a safe and efficient method of treating urinary calculi in children, which was once thought too injurious a process with first generation machines. The rapidly advancing field of laparoscopy, aided by the development of more optically refined and diminutive instruments, has allowed for its application in a wide variety of surgical interventions in pediatric urology. The tubularized incised plate urethroplasty has challenged more traditional approaches to hypospadias repair and is now considered by many pediatric urologists to be the best approach for midshaft and distal hypospadias. The one stage approach to exstrophy repair may hold the answer to improved continence without a formal bladder neck reconstruction. Finally, the field of tissue engineering leads the way to new advances in autologous biological substitutes in the surgically-challenged patient where there is a shortage of local tissues at the surgeon's disposal. PMID- 11734696 TI - Andropause: coming of age for an old concept? AB - Sex hormones have a broad range of actions in regulating very diverse systems through life as well as critical reproductive and growth processes. Sex hormone biology in its satisfaction of the early demands of species survival and reproductive advantage may be leading a destructive process resulting in frailty and the less desirable aspects of aging that may, in men, be termed andropause. One important system associated directly with aging is interleukin-6, which increases as androgens decline. This may be taking place regardless of androgen receptor activity. It is currently acknowledged that androgens are the first but not the only possible treatment for andropause. There is an acute appreciation of the potentially undesirable impact of androgens on the biology of prostate cancer, as well as, possibly, the cardiovascular system. Most authors agree that careful evaluation and surveillance of the prostate must attend androgen therapy in aging men. PMID- 11734697 TI - Update on female sexual function. AB - In this review, we briefly discuss recently published data on female sexual desire, arousal, orgasm and pain, and on medical/iatrogenic factors associated with female sexual function. The studies reviewed highlight a number of important methodological and etiological issues in the study of female sexual function. Researchers are urged to use standardized methods for defining sexual disorders and for selecting patient samples. Placebo-controlled studies are essential for examining the pharmacological aspects of female sexual dysfunction. Evidence suggests that free testosterone levels may be associated with sexual desire in women. Sildenafil citrate increases genital blood flow but may not impact on subjective reports of arousal. Past research implicated the serotonin 5 hydroxytryptamine 2 and 5-hydroxytryptamine 1A receptors in female sexual function, while recent data suggest a role for the 5-hydroxytryptamine 3 receptor. Increasing attention is being paid to medical/health conditions that impact sexual function (e.g. neurological conditions, cancer, hysterectomy, and cardiovascular disease). PMID- 11734698 TI - Counselling the patient with prostate cancer about treatment-related erectile dysfunction. AB - At present, a significant proportion of patients with prostate cancer are diagnosed at an early stage and may receive treatments able to bring about the long-term control of the disease. Thus, the impact of available treatments on the patient's quality of life has been gaining increasing importance; for patients with prostate cancer, counselling on the treatment-related effects on sexual function has become mandatory. Radical prostatectomy is very frequently performed in patients with clinically localized prostate cancer. Postoperative erectile function has been reported as being satisfactory in the majority of the patients operated on in centers of excellence for this procedure. However, overall, the results for postoperative potency are disappointing in view of the large amounts of data available from community practices. Attempts to improve postoperative potency include the intraoperative use of cavernous nerve stimulation and grafting of peripheral nerves to restore the innervation of the corpora cavernosa. Erectile dysfunction has also been associated with prostate radiotherapy. It has been shown that both ultrasound-guided brachytherapy and three-dimensional conformal radiation therapy cause an impairment of erectile function that is usually seen some time after the completion of therapy. Treatment with sildenafil citrate remains a viable option both for patients treated with radical prostatectomy (in whom the cavernous nerve function is at least partially present) and in patients treated with radiotherapy. PMID- 11734699 TI - Long-term outcomes of inflatable penile implants: reliability, patient satisfaction and complication management. AB - Despite the revolutionary introduction of oral erectogenic agents for the treatment of erectile dysfunction, there will always be patients who do not respond to conservative therapy. Penile prosthetic surgery remains an important option for these patients. Mechanical reliability and patient satisfaction have improved significantly throughout the years. This review focuses on the most recent and important updates regarding product enhancements, patient satisfaction studies, and management of intra- and postoperative problems. PMID- 11734700 TI - Modern pharmacotherapy for erectile dysfunction: evolving concepts with central and peripheral acting agents. AB - Pharmacotherapy for men experiencing erectile dysfunction has undergone dramatic advances over the past 5 years with the introduction of an effective oral agent. Sildenafil has increased the pool of couples seeking treatment for this important health issue as well as expanding the numbers of physicians treating it. Research into the growing field of erectile dysfunction is expanding at a rapid pace. Independent investigators worldwide now regularly contribute to our body of scientific knowledge. Novel oral therapies targeted at specific points along the erectile cascade are undergoing pre-clinical and early phase registration trials with the promise of rapid action, extended duration of responsiveness and an improved side effect profile. In this review, we have highlighted recent information on the next generation of phosphodiesterase inhibitors and summarized the evolving research into centrally acting agents, which may lead to effective combination therapy. PMID- 11734701 TI - New advances in Peyronie's disease. AB - The formation of Peyronie's disease plaques is a process that seems to involve a cascade of genetic, structural and immunologic events. Clinical manifestations include penile deformity and possible erectile dysfunction. Rational strategies have been forthcoming, with both minimally invasive and surgical treatments of Peyronie's disease available. This article reviews and updates current scientific and clinical advances in Peyronie's disease. PMID- 11734703 TI - DENOUEMENT--Continued from p. 1006. PMID- 11734702 TI - Erratum. PMID- 11734704 TI - Microbiology of acute otitis media recently treated with aminopenicillins. AB - INTRODUCTION: Sparse recent data are available in the United States regarding the pathogens of acute otitis media (AOM) most likely to be recovered from children recently treated with the two most frequently prescribed antibiotics, amoxicillin or amoxicillin/clavulanate (AMC). METHODS: Of the 704 rural Kentucky children with culture-positive AOM who underwent a single tympanocentesis or culture of otorrhea between 1992 and 1998, 96 pathogens were recovered from 90 children during therapy or within 7 days posttherapy with an aminopenicillin. Identification and susceptibility testing of AOM pathogens were performed by routine National Committee for Clinical Laboratory Standards methods. RESULTS: Pathogens recovered from children with AOM recently treated (0 to 7 days) with amoxicillin (n = 38) and AMC (n = 58), respectively, were as follows: Haemophilus influenzae (beta-lactamase-negative), 16 and 29%; H. influenzae (beta-lactamase positive), 11 and 22%; penicillin-susceptible Streptococcus pneumoniae, 26 and 12%; intermediately penicillin-nonsusceptible S. pneumoniae (PNSP), 20 and 10%; resistant PNSP 13 and 17%; Moraxella catarrhalis (beta-lactamase-positive), 13 and 7%; and Streptococcus pyogenes, 3 and 2%. H. influenzae was also isolated from 8 (75%) of 12 children treated with high dose AMC ( approximately 80 mg/kg/day amoxicillin component). Significantly fewer children recently treated with amoxicillin were otitis-prone than those given AMC (24% vs. 74%, P < 0.0001). CONCLUSIONS: The predominant pathogen recovered from children with AOM recently treated with amoxicillin was S. pneumoniae (59%) rather than beta lactamase-producing organisms (24%). H. influenzae was the predominant (51%) pathogen, rather than PNSP (27%), recovered from children recently treated with AMC. PMID- 11734705 TI - Pneumococcal carriage in children during their first two years: important role of family exposure. AB - BACKGROUND: Close family and day-care contacts have been identified as risk factors for pneumococcal (Pnc) carriage. This study addresses the risk of Pnc carriage by infants 2 to 24 months of age in terms of simultaneous carriage of pneumococcus by family members. METHODS: Nasopharyngeal swabs were collected from 100 Finnish infants and their family members on 10 scheduled visits (when infant was 2, 3, 4, 5, 6, 9, 12, 15, 18 and 24 months of age). The 7 most common pneumococcal serogroups (4, 6, 9, 14, 18, 19, 23), also represented in the new heptavalent Pnc conjugate vaccine, were analyzed. Marginal logistic regression analyses were performed to assess the relative importance of different predictors for carriage. RESULTS: The carriage of the studied Pnc serogroups increased with age, being highest at the age of 18 months (28%). Among children older than 6 months of age, the strongest predictor of carriage was simultaneous carriage of the same serogroup by another family member (odds ratio, 3.8; 95% confidence interval, 2.1 to 6.9). At the age of 6 months or younger, carriage was rare and was not significantly associated with a family carriage. CONCLUSIONS: Young infants (< or =6 months old) were largely protected from pneumococcal carriage. After this age family transmission seemed very important despite the small family size. Contrary to some earlier studies communal day care was not associated with an increased risk of Pnc carriage. This could be partly because of the long parental leave in Finland and thus the late age of starting organized day care. PMID- 11734706 TI - Clinical profile of serologically diagnosed pneumococcal pneumonia. AB - OBJECTIVE: To describe the characteristics of serologically diagnosed pneumococcal pneumonia and compare them with those of respiratory syncytial virus (RSV) pneumonia and bacteremic pneumococcal pneumonia. METHODS: IgG antibodies to pneumococcal pneumolysin and C-polysaccharide as well as immune complexes containing IgG antibodies to pneumolysin and C-polysaccharide were measured from acute and convalescent sera of 254 children with community-acquired pneumonia. Evidence of pneumococcal infection was found in 93 children. Clinical and laboratory data were retrospectively collected from the records of 38 children with sole (all tests for 16 other microbes negative) pneumococcal pneumonia and compared with 26 sole RSV-induced pneumonia from the present series and with the data of our 85 bacteremic pneumococcal pneumonia cases reported earlier. RESULTS: Serologically diagnosed sole pneumococcal pneumonia clinically overlapped with RSV pneumonia, but RSV pneumonia was more often associated with tachypnea (45% vs. 17%, P < 0.05) and low white blood cell counts (means, 12.0 x 109/l vs. 20.8 x 109/l; P < 0.001) as well as low serum C-reactive protein levels (means, 28 mg/l vs. 137 mg/l; P < 0.001). Alveolar infiltrates were found in 15% of chest radiographs of children with RSV pneumonia compared with 76% of those in children with sole pneumococcal pneumonia (P < 0.001). Patients with bacteremic pneumonia more often appeared ill (79% vs. 50%, P < 0.001) and more often had typical pneumococcal pneumonia with high fever, leukocytosis and lobar infiltrates in their chest radiographs (70% vs. 34%, P < 0.05) than those with serologically diagnosed pneumococcal pneumonia. CONCLUSIONS: Serologically detected pneumococcal pneumonia differs significantly from RSV pneumonia in laboratory and chest radiography findings, but the clinical signs and symptoms overlap considerably. Bacteremic pneumococcal pneumonia is a more severe illness than the serologically diagnosed one. PMID- 11734707 TI - Twelve year observation of primary and secondary antibiotic-resistant Helicobacter pylori strains in children. AB - BACKGROUND: The effectiveness of Helicobacter pylori eradication regimens is influenced by antibiotic susceptibility of infecting strains. Data concerning antibiotic resistance in children are limited. We report the evolution of primary and secondary resistance in a series of Belgian children during the last 12 years. PATIENTS AND METHODS: From 1989 through 2000, H. pylori gastritis was diagnosed in 569 children, and antibiotic susceptibility tests were performed in 555. Eradication, using different schemes, failed in 128 of 457 treated children. After eradication failure antibiotic susceptibility determination was performed in 87 of 128. Comparison of antibiotic susceptibility of strains isolated from the gastric body and from the antrum was performed in 238 samples. RESULTS: Resistance to amoxicillin was not observed. The rate of primary resistance to nitroimidazole derivatives was 18.0% (101 of 555) and remained constant throughout this period, whereas primary resistance to macrolides increased from an average of 6.0% (range, 0 to 10%) before 1995 to 16.6% (range, 10 to 25%, P < 0.001) thereafter. Antibiotic consumption in Belgium, especially macrolides, did not show important fluctuations during the study period. Secondary resistance developed in 39 of 87 patients (46%). Strains isolated from different gastric locations show identical susceptibility testing in all but 5 of 238. CONCLUSIONS: Resistance of H. pylori to macrolides increased in our pediatric population which did not appear to correlate with macrolides prescription habits in our country. After eradication failure acquired secondary resistance was observed in one-half of the patients. PMID- 11734709 TI - Epidemiologic aspects and preventive strategy of hepatitis B and C viral infections in children with cancer. AB - AIMS: We present the efficacy of a strategy to control infections with hepatitis B (HBV) and C viruses (HCV) in children with cancer and assessment of risk for their relatives and health care personnel. A total of 1242 people entered the study, including 558 children with cancer, 193 relatives of infected children, 302 health care workers and 189 controls. METHODS: To stop dual HBV and HCV nosocomial infection in the oncology department, a preventive strategy was introduced. It involved immunoprophylaxis against HBV, screening blood donors for HCV infection, intensification of nonspecific prophylaxis, an educational program and estimation of risk for relatives of infected children and health care personnel. RESULTS: Retrospective analysis showed that the prevalence of HBV and HCV infections in children with cancer was 74 of 119 (62.2%) and 50 of 92 (54.3%), respectively, with the highest rate among patients with leukemia. Inferior anticancer therapeutic response were obtained in infected children. Specific anti-HBV immunoprophylaxis introduced simultaneously with anticancer therapy resulted in protection of 160 of 168 (95.2%) children in the first 4 years, when 62.9% of patients receiving therapy developed protective antibodies. Screening of blood donors and intensification of nonspecific prophylaxis reduced HCV prevalence to 2.8% during the most recent 1.5 years. Genotype analysis showed that the risk of HCV infection was 0.5% for relatives of infected children. The risk for health care personnel was 0 in the oncology ward and 1.9% in the other departments, and it reached 0.53% in control group. CONCLUSIONS: The preventive strategy of viral hepatitis in children with cancer, including passive-active HBV immunoprophylaxis from the beginning of chemotherapy and intensive nonspecific prophylactic measures is effective. With this strategy the risk of intrafamily and occupational infection is low. PMID- 11734708 TI - Short term oral cefixime therapy for treatment of bacterial conjunctivitis. AB - BACKGROUND: There have been few controlled studies evaluating treatment of bacterial conjunctivitis beyond the newborn period. Topical therapy of bacterial conjunctivitis achieves a clinical cure but does not prevent acute otitis media (AOM). OBJECTIVES: The aim of this study was to compare systemic antibiotic therapy (cefixime) with topical therapy with polymyxin-bacitracin for treatment of acute bacterial conjunctivitis with regard to clinical and bacteriologic cure and prevention of AOM. METHODS: This study was a randomized, double blind, placebo-controlled trial of polymyxin-bacitracin ointment and oral placebo vs. topical placebo and oral cefixime in children with presumed acute bacterial conjunctivitis. Topical therapy was administered for 7 days; oral therapy was administered for 3 days. Bacterial cultures were obtained at entry and on Day 3 of treatment. Children were examined on Days 3 and 10 or if they worsened within 15 days of entry. RESULTS: Eighty children were enrolled in the study. Bacterial cultures of the conjunctiva were positive in 70% of children: Haemophilus influenzae (53.7%); Streptococcus pneumoniae (13.8%); H. influenzae and S. pneumoniae (1.2%); and Moraxella catarrhalis (1.3%). There were 7 (17.5%) bacteriologic failures among children receiving topical antibiotic and oral placebo and 15 (37.5%) bacteriologic failures among children receiving topical placebo and oral cefixime (P = 0.07 with Yates correction). There was no difference between study groups with regard to either clinical cure or the development of AOM. Nine children (11%), 5 who received active topical therapy and 4 who received active oral drug, developed AOM either during or within 15 days of study entry. CONCLUSION: Cefixime was not more effective than topical polymyxin-bacitracin in either the eradication of conjunctival colonization with respiratory pathogens or the prevention of AOM in children with acute bacterial conjunctivitis. PMID- 11734710 TI - Safety of neonatal hepatitis B vaccine administration. AB - OBJECTIVE: To determine whether hepatitis B vaccination of newborns increases the incidence of fever and/or suspected sepsis. METHODS: A prospective clinical study was undertaken at the Kaiser Permanente San Francisco Medical Center involving normal full term newborns born between November 1, 1991, and April 30, 1994. During this time 3302 infants were vaccinated within 21 days of birth with hepatitis B vaccine, and 2353 were not. Clinical and demographic data were collected from Kaiser Permanente's existing clinical information systems, and laboratory data for blood and cerebrospinal fluid (CSF) cultures were obtained from the comprehensive automated regional laboratory reporting system. RESULTS: There were no significant differences between vaccinated and unvaccinated newborns in the proportion of infants who received care for fever (0.8% vaccinated and 1.1% unvaccinated, P = 0.28), allergic reactions, seizures or other neurologic events in the first 21 days of life. Vaccinated newborns were significantly less likely to undergo microbiologic evaluation for possible sepsis. Among vaccinated newborns 4.0% had blood cultures and 1.6% had CSF cultures. Among infants who were not vaccinated 8.3% had blood cultures and 1.6% had CSF cultures (P <0.001 for both tests). CONCLUSION: This study found no evidence that newborn hepatitis B vaccination is associated with an increase in the number of febrile episodes, sepsis evaluations or allergic or neurologic events. In addition our data did not support any increase in medical procedures attributed to receipt of hepatitis B vaccine. PMID- 11734711 TI - Immunogenicity of a serogroup B meningococcal vaccine against multiple Neisseria meningitidis strains in infants. AB - BACKGROUND: The serogroup B meningococcus is responsible for the majority of cases of meningococcal disease in temperate countries. Infants and young children <2 years of age are at greatest risk of disease. This study assessed the immunogenicity in infants of a serogroup B meningococcal outer membrane protein vaccine that has been used extensively in disease outbreaks in Cuba and several Latin American countries and shown to be efficacious in teenagers. METHOD: One hundred five healthy infants entering the routine vaccination schedule in Havana, Cuba, were given either 2 or 3 doses of the serogroup B meningococcal vaccine VA MENGOC-BC at 3.5, 5.5 and 7.5 months of age. Immune response pre- and postvaccination was determined by the conventional serum bactericidal assay (SBA), a more sensitive novel whole blood bactericidal assay (WBA) and immunoglobulin ELISA. RESULTS: In 52 and 46% of infants >50% killing of the vaccine serogroup B strain (B:4:P1.19,15) and serogroup C strain, respectively, was demonstrated by the WBA after 2 doses of the vaccine. Serum bactericidal activity (4-fold increase in titer) was induced in only 27% against the vaccine serogroup B strain and in 14% against the serogroup C strain. The changes in WBA and SBA were mirrored by the serogroup B and C immunoglobulin ELISA. Cross reactive immunogenicity against other (heterologous) serogroup B strains was demonstrated for one of the four further strains assessed by WBA. By age 16 to 18 months SBA, WBA and ELISA responses had declined considerably. The addition of a third dose of vaccine did not appear to significantly influence immunogenicity at 17 months of age. CONCLUSION: The serogroup B outer membrane protein vaccine VA MENGOC-BC induces a demonstrable immune response in infants against both the serogroup B vaccine strain and against a serogroup C strain. Cross-reactive immunogenicity against other (heterologous) serogroup B strains is limited in this age group. PMID- 11734712 TI - Use of purified protein derivative to assess the risk of infection in children in close contact with adults with tuberculosis in a population with high Calmette Guerin bacillus coverage. AB - SETTING: Household contacts <15 years of age of adults with tuberculosis (TB) attending a reference center in Aracaju, Sergipe, Northeast Brazil. OBJECTIVE: To assess the use of purified protein derivative (PPD) and frequency of infection in children with high Calmette-Guerin bacillus (BCG) coverage who were recently exposed to TB. DESIGN: Cross-sectional study of 141 exposed household contacts <15 years of age and 506 nonexposed neighborhood controls. Children were examined and assessed for degree of exposure to index cases, BCG vaccination and scar and were tested with PPD. RESULTS: Exposed children were younger and less up to date in their vaccination schedule than controls (P < 0.05). BCG had been given to 95.6% of exposed children and 97.4% of controls, but only 80.9% of exposed vs. 88.5% of controls had a scar (P < 0.05). Scar sizes of exposed children were smaller (medians, 4.5 and 7 mm, respectively; P < 0.05). Children had lower weight for age z scores and height than the National Center for Health Statistics standards. Exposed children had lower weight for age z scores than controls (P < 0.05). Sixty-seven (47.5%) exposed children and 18 (3.6%) controls had PPD readings of >10 mm. Positivity and induration sizes increased with age, although this was significant only in the controls. The presence of a BCG scar was not associated with having a positive PPD. The degree of exposure was an important factor for PPD positivity; 66 (60.6%) of the 109 children with close exposure were positive compared with 1 (3.1%) of 32 with less intimate exposure. PPD indurations among close contacts were also larger than those with restricted exposure and controls (16.3, 11 and 9.4 mm, respectively; P < 0.05). PPD positivity was associated with the quantification of acid-fast bacilli in the adult; 38.5% of contacts with adults with sputum with (+) were positive, compared with 42.3% of those with (++) and 58.4% with (+++) (P < 0.05). CONCLUSION: PPD is a useful test to identify children infected with TB independently of whether they had received BCG or not. Children exposed to adults with TB are at high risk of infection. The risk of infection is associated with the intimacy of contact and the number of bacilli expectorated in sputum. PMID- 11734713 TI - Risk factors for human immunodeficiency virus infection in Ethiopian children with tuberculosis. AB - BACKGROUND: Separate risk factors for HIV infection and for tuberculosis (TB) are well-studied, but it is unclear whether these risk factors still apply in the new epidemiologic situation of dual infection. This study examines risk factors associated with seropositivity for HIV in Ethiopian children with clinical TB. METHODS: A prospective, controlled study of children with TB diagnosed in Addis Ababa from December 11, 1995, to January 28, 1997, in which HIV-positive children were compared with HIV-negative children with regard to sociodemographic background, previous medical history and vaccination. RESULTS: HIV prevalence among children with clinical TB was 11.2%. High educational status of mothers, low age, loss of one or two parents and earlier Calmette-Guerin bacillus (BCG) vaccination of the child were factors independently related to HIV infection. CONCLUSION: Factors associated with HIV infection among children with clinical TB include higher education of parents, higher income and better living conditions. The HIV epidemic might thus modify traditional risk factors for tuberculosis. It might also decrease the overall effect of BCG vaccination given that BCG did not provide protection in children infected with HIV. An expected increase of dually infected children who are younger, more in need of hospitalization and often lacking one or both parents will put an additional burden on the Ethiopian health care system. PMID- 11734714 TI - John Snow learns from Louis Pasteur. PMID- 11734715 TI - Pediatric uses of valacyclovir, penciclovir and famciclovir. PMID- 11734716 TI - Poststreptococcal reactive arthritis. PMID- 11734717 TI - Successful treatment of foscarnet-resistant herpes simplex stomatitis with intravenous cidofovir in a child. AB - A leukemic child developed recurrent herpes simplex virus lesions shortly after receiving a bone marrow transplant and while taking acyclovir. The isolate was resistant to acyclovir and foscarnet in vitro. The lesions responded to a course of cidofovir. PMID- 11734718 TI - Most ten-year-old children with negative or unknown histories of chickenpox are immune. AB - To evaluate the proportion of children to vaccinate against varicella in a catch up program targeting 9- to 10-year-old children, a study was conducted among children age 10 years to assess the age-specific incidence of varicella and document the immunity against varicella in those with negative or unknown chickenpox history. Of the latter 62% were seropositive for varicella. PMID- 11734719 TI - Detection of group C rotavirus in children with acute gastroenteritis in Blantyre, Malawi. AB - Among 606 children who were treated for acute gastroenteritis at the Queen Elizabeth Central Hospital in Blantyre, Malawi, Group C rotavirus (Gp C RV) was detected by enzyme-linked immunosorbent assay in fecal specimens from 16 (3.9%) of 408 inpatients and in 4 (2.0%) of 198 outpatients. Thirteen (65%) children excreting Gp C RV were coinfected with Group A rotavirus. PMID- 11734720 TI - Experience with routine voluntary perinatal human immunodeficiency virus testing in an inner city hospital. AB - Women enrolled in prenatal care at Grady Health System, Atlanta, GA, have routinely been offered HIV counseling and voluntary testing since 1987. Consistently >90% have accepted testing. With implementation of US Public Health Service guidelines for perinatal zidovudine prophylaxis in 1994, the mother-to child HIV transmission rate rapidly decreased from 18% to 8% during the subsequent 2 years. PMID- 11734721 TI - Early onset pneumococcal sepsis in children hospitalized for noninfectious life threatening events. AB - During a 13-year period 9 patients admitted to a pediatric intensive care unit for life-threatening noninfectious conditions developed pneumococcal sepsis within 48 h of admission. All patients were Bedouins, a population group characterized by high prevalence of respiratory carriage of Streptococcus pneumoniae. In populations with high carriage rates of S. pneumoniae, critically ill children appear to be at increased risk of pneumococcal sepsis. PMID- 11734722 TI - Development of acyclovir-resistant herpes simplex virus early during the treatment of herpes neonatorum. AB - Genotypic analysis of herpes simplex virus (HSV) DNA extracted from clinical specimens from a case of fatal disseminated neonatal HSV demonstrated that an infant developed an acyclovir-resistant HSV containing a mutation in the HSV thymidine kinase gene during the first seven days of acyclovir therapy. PMID- 11734723 TI - Thorn-induced Alternaria flexor tenosynovitis of the hand. AB - A thorn puncture to the hand is a common, usually minor childhood injury. A 6 year-old boy developed Alternaria tenosynovitis as a complication of thorn penetration into the digital flexor tendon sheath. He was successfully treated with tenosynovectomy and a prolonged course of itraconazole. PMID- 11734724 TI - Nodular rash in an Indonesian girl. PMID- 11734725 TI - Chemoprophylaxis for household contacts of index cases of invasive non-type b Haemophilus influenzae disease. PMID- 11734726 TI - Hospitalization costs of respiratory syncytial virus infection. PMID- 11734727 TI - False positive hepatitis B surface antigen test caused by hepatitis B vaccine. PMID- 11734729 TI - DENOUEMENT Continued from p. 554. PMID- 11734730 TI - DENOUEMENT-Continued from p. 724. PMID- 11734731 TI - The new nasal spray influenza vaccine. PMID- 11734733 TI - Efficacy of inactivated and cold-adapted vaccines against influenza A infection, 1985 to 1990: the pediatric experience. AB - BACKGROUND: Influenza is a common and potentially serious infection in children. Although there is interest in broadening the use of influenza vaccine in healthy children, there are few large, randomized, controlled trials that evaluate the safety and efficacy of inactivated vaccine in the pediatric population. METHODS: From 1985 through 1990 a randomized, controlled trial of cold-adapted and inactivated vaccines for the prevention of influenza A disease was conducted at Vanderbilt University, and the cumulative results from this trial in patients of all ages have been previously published. We reanalyzed the data from this trial in the subset of patients who were younger than 16 years at the time of their participation. We determined vaccine safety, immunogenicity and efficacy, based on culture-positive illness and seroconversion, in this subset of patients. RESULTS: During the 5 years of the study, 791 children younger than 16 years received 1809 doses of either inactivated or cold-adapted vaccine or placebo. The vaccines were well-tolerated, and there were no serious reactions. Inactivated trivalent influenza vaccines were 91.4 and 77.3% efficacious in preventing symptomatic, culture-positive influenza A H1N1 and H3N2 illness, respectively. The efficacy of the inactivated vaccine based on hemagglutination inhibition assay seroconversion was 67.1 and 65.5%, respectively, for H1N1 and H3N2 serotypes. CONCLUSIONS: Inactivated trivalent influenza A vaccines are well tolerated and efficacious in the prevention of influenza A disease in children 1 to 16 years old. PMID- 11734734 TI - Prospective, randomized, placebo-controlled evaluation of the safety and immunogenicity of three lots of intranasal trivalent influenza vaccine among young children. AB - BACKGROUND: Trivalent formulations of an experimental, cold-adapted, intranasal influenza (CAIV) vaccine have been shown to be safe, immunogenic and efficacious in young children. METHODS: We evaluated the safety and immunogenicity of three consistency lots of CAIV in children 12 to 36 months of age randomized to one of five groups: Groups 1, 2 and 3 received separate lots containing A/Shenzhen/227/95 (H1N1), A/Wuhan/359/95(H3N2) and B/Harbin/7/94-like viral strains. Group 4 received an earlier efficacy trial lot which included a different H1N1 strain (A/Texas/36/91-like); and Group 5 received placebo. We performed strain-specific serum hemagglutination inhibition antibody levels against type A (H3N2 or H1N1) or type B as appropriate. RESULTS: Overall 474 children received 2 doses, 2 months apart. Each lot was well-tolerated, and there were no significant group differences between consistency lots in the proportion of children with fever and local or systemic reactions after vaccination. The 3 consistency lots were not statistically different with regard to immunogenicity as measured by seroconversion or absolute geometric mean titer. Immune responses were more robust among initially seronegative children and for H3N2 and B strains than for H1N1 strains. After 2 doses of vaccine 97, 84 and 62% had hemagglutination inhibition titers > or = 1/32 against A/H3N2, B and H1N1 strains, respectively. For A/H3N2 only, immune responses after 1 dose of vaccine are similar to those seen after 2 doses. CONCLUSIONS: Each consistency lot of CAIV is as or more immunogenic than a lot used in a large efficacy trial. PMID- 11734737 TI - Prognosis for neonates with enterovirus hepatitis and coagulopathy. AB - BACKGROUND: Neonatal enterovirus hepatitis and coagulopathy (EHC) can be a severe, life-threatening infection. However, the case-fatality rate of EHC and the prognosis for survivors are not well-defined. METHODS: A search of a hospital medical records database and the investigator's files for the period 1983 to 2000 was performed. Patients with onset of enterovirus-associated illness at age < or = 30 days and either (1) aspartate aminotransferase or alanine aminotransferase > 3 times the upper limit of normal or (2) platelet count < 100 000 plus an abnormal coagulation profile were included. RESULTS: Sixteen cases were identified. Symptoms began at a mean of 3.8 days (range, 1 to 7 days). Frequent clinical and laboratory findings included jaundice, lethargy, anorexia, hepatomegaly, thrombocytopenia, clotting time prolongation, elevated transaminase and bilirubin values and decreased fibrinogen and albumin concentrations. Five patients had myocarditis and 4 had encephalitis. Hemorrhagic complications occurred in 10 (intracranial hemorrhage in 5). Five (31%) patients died. Features discriminating patients who died from survivors were concomitant myocarditis (5 of 5 vs. 0 of 11, P < 0.001), encephalitis (3 of 5 vs. 1 of 11, P = 0.06), prothrombin time > 30 s (4 of 5 vs. 1 of 9, P = 0.02) and intracranial hemorrhage (4 of 5 vs. 1 of 8, P = 0.03). Follow-up of 6 survivors revealed normalization of liver function and platelet counts, satisfactory growth and absence of significant medical problems. CONCLUSIONS: The case-fatality rate of 31% is evidence of the potentially devastating nature of EHC. Concomitant EHC and myocarditis are especially serious, but the prognosis for children who survive neonatal EHC is generally excellent. PMID- 11734736 TI - Appropriateness of antibiotic prescriptions in community-acquired acute pediatric respiratory infections in Spanish emergency rooms. AB - OBJECTIVE: To describe the variability and appropriateness of antibiotic prescriptions in community-acquired acute respiratory infections (ARI) during childhood in Spain. METHODS: A descriptive, multicenter study of variability in clinical practice was conducted by evaluating a prospective series of pediatric patients attending the emergency rooms of 11 Spanish hospitals and diagnosed with community-acquired ARI. The appropriateness of the antibiotic prescriptions was assessed by comparing our clinical practice with consensus guidelines developed for this study. RESULTS: We collected data from 6,249 ARI emergencies studied on 30 separate days. Antibiotics were prescribed in 58.7% of the ARI (bronchiolitis, 11.5%; bronchitis, 40.2%; pharyngotonsillitis, 80.9%; nonspecified ARI, 34.8%; pneumonia, 92.4%; otitis, 93.4%; sinusitis, 92.6%). The most commonly used antibiotics were amoxicillin/clavulanate (33.2%), amoxicillin (30.2%), cefuroxime axetil (8.5%) and azithromycin (6%). According to the consensus guidelines developed for this study, therapy was considered to be appropriate in 63.1% of the ARI (first choice, 52.1%; alternative choice, 11.0%) and inappropriate in 36.9%. The percentages of inappropriate prescription according to ARI groups were: bronchiolitis, 11.5%; bronchitis, 31.5%; pharyngotonsillitis, 54.8%; nonspecified ARI, 34.7%; pneumonia, 13.9%; otitis, 25.6%; and sinusitis, 22.2%. CONCLUSIONS: There is excessive use of antibiotics in acute respiratory infections that are presumably viral in origin. An important number of ARI of potentially bacterial origin are treated with antibiotics that are not sufficiently efficacious or that have a broader spectrum than necessary. PMID- 11734735 TI - Pharmacokinetics of nelfinavir in human immunodeficiency virus-infected infants. AB - BACKGROUND: Nelfinavir dosed at approximately 20 to 30 mg/kg three times a day (TID) in older children provides exposure similar to 750 mg TID in adults. However, the pharmacokinetics (PK) of nelfinavir in infants who are < 2 years of age is not well-described. The objective of this study was to determine the pharmacokinetics of nelfinavir in infants < 2 years of age. METHODS: Nelfinavir concentrations were evaluated in 22 HIV-infected infants between 15 days and 2 years of age receiving nelfinavir as part of Pediatric ACTG Study 356. Nelfinavir therapy was initiated at approximately 25 mg/kg TID (n = 18) or approximately 55 mg/kg twice a day (n = 4) and given in combination with nevirapine, stavudine and lamivudine. PK samples were obtained predose and 1.5 and 4 h postdose at approximately 6-month intervals. Eight infants (all < or = 3 months of age) also had intensive PK samples collected at Week 1. RESULTS: The median apparent clearance in the infants with intensive pharmacokinetic sampling was 2.7 liters/h/kg (range, 1.8 to > or = 10) and was similar between twice a day and TID dosing cohorts. Overall nelfinavir concentrations at all collection times were lower in these infants than previously reported in older pediatric patients. CONCLUSIONS: Nelfinavir concentrations in infants are highly variable and lower than those seen in adult or older pediatric populations receiving labeled dosing. Therefore it is necessary to further evaluate nelfinavir safety, effectiveness and pharmacokinetics at higher doses than used among other pediatric populations. PMID- 11734738 TI - Community-acquired methicillin-resistant Staphylococcus aureus colonization in healthy children attending an outpatient pediatric clinic. AB - BACKGROUND: We previously showed that children attending an inner city pediatric emergency department were sometimes asymptomatically colonized with clindamycin susceptible community-acquired methicillin-resistant Staphylococcus aureus (MRSA) and borderline methicillin-resistant S. aureus (BRSA) as well. We wished to ascertain whether healthy children attending an outpatient clinic were colonized with these organisms. Therefore to estimate the prevalence of community-acquired MRSA and BRSA nasal colonization in a well child population, we cultured children attending an inner city pediatric outpatient clinic. STUDY DESIGN: This was a prospective cross-sectional study conducted from January to August, 1999, at a primary care outpatient facility at the University of Chicago. The target population was 500 healthy children < or = 16 years of age who attended this facility to receive well child care. RESULTS: One hundred twenty-two (24.4%) children were colonized with S. aureus. Three of the 122 (2.5%) S. aureus isolates were MRSA; they came from children who lacked predisposing risk factors and were susceptible to clindamycin, gentamicin, trimethoprim-sulfamethoxazole, rifampin and ciprofloxacin. Two (1.6%) additional S. aureus isolates were BRSA; both children had predisposing risk factors for MRSA colonization. The mecA gene was present in the 3 MRSA isolates and absent in both BRSA isolates. CONCLUSIONS: These data document that a reservoir of asymptomatic MRSA colonization exists among healthy children who lack traditional risk factors for MRSA infections. PMID- 11734739 TI - Haemophilus influenzae type b disease: impact and effectiveness of diphtheria tetanus toxoids-acellular pertussis (-inactivated poliovirus)/H. influenzae type b combination vaccines. AB - BACKGROUND: Since 1996 in Germany primary infant immunization against Haemophilus influenzae has been most commonly given in the form of diphtheria-tetanus toxoids acellular pertussis/H. influenzae type b (DTaP/Hib) or diphtheria-tetanus toxoids acellular pertussis (-inactivated poliovirus)/H. influenzae type b (DTaP-IPV/Hib) combination vaccines. These combination vaccines elicit lower anti-Hib antibody concentrations than the equivalent Hib conjugate administered as a separate injection, but the clinical relevance of this phenomenon is unknown. METHODS AND FINDINGS: To assess the impact of DTaP/Hib combination vaccines on the incidence of invasive Hib disease in Germany, two independent surveillance systems, one hospital- and one laboratory-based, were used during 1998 and 1999 for detection of cases. Vaccination histories of all cases detected were obtained by telephone contact with parents or health care providers. During the 2-year study period invasive H. influenzae disease in the <5-year age group continued to fall, with a mean annual incidence of 1.01/100 000 children. National vaccination coverage rates revealed that only 70% of children given DTaP/Hib or DTaP-IPV/Hib received the recommended three doses in their first year of life, but the overall effectiveness of these vaccines was high at 97.5% (95% confidence interval, 96.3 to 98.4) for those who had received at least one dose. In subjects who received the full 3-dose schedule, effectiveness was 98.8% (95% confidence interval, 98.2 to 99.3). CONCLUSION: Although it is well-documented that DTaP/Hib vaccines elicit lower anti-Hib titers than separate vaccines, such combinations are effective in reducing the incidence of invasive H. influenzae type b disease. PMID- 11734740 TI - Seroprevalence of varicella-zoster virus immunoglobulin G antibodies in Swiss adolescents and risk factor analysis for seronegativity. AB - BACKGROUND: Little is known about the seroprevalence of anti-varicella-zoster virus (VZV) serum antibodies in adolescents in Switzerland as in most other European countries. METHODS: Serum specimens from 13- to 15-year-old students from eight urban and rural areas in Switzerland, obtained as part of an allergy risk assessment study project (SCARPOL), were available for analysis of IgG antibodies against VZV by enzyme-linked immunosorbent assay (ELISA) and confirmation by fluorescent antibody staining of membrane antigen in a subcohort. Serum specimens and comprehensive sociodemographic data had been collected during two study periods between 1992 and 1995. RESULTS: Data and serum specimens were available from 1709 and 1788 subjects, respectively. Seroprevalence of anti-VZV antibodies as measured by ELISA was 95.5% (95% confidence interval, 94.5 to 96.4). When serum specimens that were indeterminate by ELISA were tested by FAMA, seroprevalence was 96.5% (95% confidence interval, 95.7 to 97.4). After logistic regression analysis, the number of siblings was the only factor that significantly influenced the presence of VZV antibodies (90.1% in those with no siblings, >96% with 1 or more siblings), whereas residence (urban vs. rural), parental education, nationality and gender did not. CONCLUSIONS: Seroprevalence of anti-VZV serum antibodies is comparatively high among Swiss adolescents. Individuals who grow up without siblings have a significant risk of evading natural VZV infection in childhood, and they therefore form a potential target group for varicella immunization in Switzerland. PMID- 11734741 TI - Nasopharyngeal colonization by Haemophilus influenzae in children living in an orphanage. AB - AIM: To study colonization and transmission of Haemophilus influenzae in a cohort of children <2 years old living in the unique epidemiologic conditions of a closed community of an orphanage. METHODS: Fifty-three children, ages 0 to 24 months, were followed for 1 year. All children >2 months were vaccinated against H. influenzae serotype b. Nasopharyngeal cultures were collected monthly or, in children <6 months of age, every 2 weeks. Antibiotic susceptibility, serotype, biotype and genotype (pulsed field gel electrophoresis) of each isolate were determined. As control, 39 H. influenzae isolates were recovered from various regions in France. RESULTS: The mean monthly rate of carriage was 45% ranging from 17 to 70%. Most isolates belonged to biotype II (62%), 4 isolates to serotype f (3.6%) and none to serotype b, and 60% of the 111 isolates produced beta-lactamase. A complete concordance was found among biotype, serotype, pulsotype and antimicrobial susceptibility. On average children were sequentially colonized by 3 different isolates. The mean duration of carriage for a given isolate was approximately 1.4 months. In younger children the mean age of primary colonization was 2 months. Contrasting with the high genetic heterogeneity of 39 control isolates, most isolates (82%) belonged to only 5 pulsotypes. Three main H. influenzae clones rapidly spread in the community and colonized children in waves. CONCLUSION: During early life nasopharyngeal colonization by H. influenzae is a dynamic phenomenon with sequential carriage of various clones spreading in the community. PMID- 11734742 TI - International variation in disease burden of rotavirus gastroenteritis in children with community- and nosocomially acquired infection. AB - BACKGROUND: The great impact of rotavirus disease on morbidity and medical health care costs in industrialized countries together with the withdrawal of the live oral rotavirus vaccine have made a reassessment of rotavirus gastroenteritis necessary. Such a reassessment should provide sufficient data for developing alternative disease prevention strategies and for allocating resources efficiently. OBJECTIVES: To compare characteristics and management of community- and nosocomially acquired rotavirus disease in Austria, Germany and Switzerland. PATIENTS AND METHODS: In a prospective, population-based, trinational (Austria, Germany, Switzerland), multicenter (9 cities, 10 hospitals and 30 pediatric practices) study, a total of 174 552 children months and 78 516 hospital days were evaluated. Participants were all children 4 years of age and younger, who either presented at one of the pediatric practices with community-acquired gastroenteritis, or who had acquired gastroenteritis nosocomially. From December, 1997, to May, 1998, prospective antigen testing was done by enzyme-linked immunosorbent assay, and serotyping was done by reverse transcription polymerase chain reaction. Disease severity was scored by the Vesikari severity scale. RESULTS: Rotavirus was detected in 29.5, 27 and 37.5% of children with community acquired gastroenteritis and in 57, 69 and 49% of children with nosocomial gastroenteritis in Austria, Germany and Switzerland, respectively. Severity of community-acquired rotavirus gastroenteritis was more pronounced in Austria (median severity score, 11) than in Germany (median score, 9) or Switzerland (median score, 10). However, only 2% of Austrian and Swiss children compared with 12% of German children presented to their pediatricians more than four times. Nosocomially acquired rotavirus gastroenteritis was mildest in Austria but occurred within the shortest median duration of hospitalization (4 days vs. 5 and 7 in Germany and Switzerland, respectively). In a multivariant analysis age, family size, day care, breast-feeding and nationality were not predictive factors for enhanced risk to contract rotavirus infection. Alimentation was changed frequently; diet was used between 23 and 83%; special formulas were used between 10 and 57%. CONCLUSION: The cumulative experience from three European countries suggest that rotavirus is an important cause of diarrhea in Central Europe, but significant local differences clearly demonstrate the need for obtaining national data as a reliable basis for control and prevention of the disease. PMID- 11734743 TI - Asymptomatic salmonellosis among children in day-care centers in Merida, Yucatan, Mexico. AB - BACKGROUND: Child day-care centers (DCC) have become common in many lower and middle income countries, presenting new problems that may differ from those of DCC in more developed countries. Diarrhea is a common problem in DCC in the United States, but information on the prevalence of diarrhea or specific enteropathogens among children in DCC in tropical and developing countries is limited. METHODS: Because of preliminary data from newborns and DCC attendees in Merida, Mexico, with high rates of Salmonella infection, we conducted a 12-month longitudinal surveillance study of enteropathogens in two Merida DCC. Seventy eight children ages 2 months to 4 years were evaluated with demographic and clinical data, and stools were cultured monthly. RESULTS: Salmonella sp. was the most common enteropathogen detected (46 of 683 specimens, 6.7%), with higher rates in children younger than 18 months (P < 0.02), but it was found in only 1 of 10 diarrhea episodes that coincided with sampling. Other common organisms identified included Giardia lamblia (21 of 683, 3.0%) and LT-producing enterotoxigenic Escherichia coli (16 of 683, 2.3%). Salmonella was recovered from as many as 19% of children in a single month, but the large multiplicity of serotypes recovered suggested multiple sources rather than a common source outbreak. Children with Salmonella tended to have more liquid stools during the preceding 2 weeks. Salmonella was also isolated from the stool of teachers in 1 of the 2 DCC in 10 of 94 specimens (10.6%), and again multiple serotypes were represented. CONCLUSION: These data indicate the presence of multiple sources of Salmonella infection in the DCC, posing a complex situation for infection control. PMID- 11734744 TI - Bacterial endocarditis in infants and children. PMID- 11734745 TI - West Nile virus. PMID- 11734746 TI - Nevirapine pharmacokinetics in pregnant women and in their infants after in utero exposure. AB - The safety, toxicity and pharmacokinetics of nevirapine were studied in HIV infected pregnant women beginning chronic therapy late in the third trimester and in their infants. Initial dose pharmacokinetic profiles in the pregnant women were similar to those seen in nonpregnant adults. Serum nevirapine concentrations fell below the 100-ng/ml target concentration by Day 7 of life in four infants, suggesting that nevirapine elimination is accelerated in these infants compared with newborns whose mothers receive only a single intrapartum nevirapine dose. PMID- 11734747 TI - Hepatitis A vaccine in pediatric patients affected by metabolic liver diseases. AB - Twenty children with a variety of metabolic liver diseases were given two doses of hepatitis A vaccine. Adverse reactions were mild. All subjects responded to vaccine with seroconversion to hepatitis A virus antibodies after the first dose, regardless of transaminase values, and had a booster effect from the second doses. PMID- 11734748 TI - Outbreak of scarlet fever caused by an erythromycin-resistant Streptococcus pyogenes emm22 genotype strain in a day-care center. AB - We report an outbreak of scarlet fever and pharyngeal colonization with Streptococcus pyogenes in a day-care center in Mexico City. The outbreak strain was resistant to erythromycin but susceptible to clindamycin. T-type 11,12 serotype was found in eight isolates, from two patients and six carriers, which had the emm22 gene. The recognition of streptococci resistant to macrolides causing outbreaks has implications for infection control and for improving antibiotic prescribing patterns in the day-care setting. PMID- 11734749 TI - Autoimmune complications associated with hepatitis A virus infection in children. AB - Hepatitis A virus infection is usually a self-limited disease during childhood. Autoimmune manifestations are rarely reported among patients. We describe two children with acute hepatitis A infection who developed immune thrombocytopenia and hepatic venous thrombosis during the course of acute infection. Antiphospholipid antibodies were increased in both of them during the thrombocytopenic and thrombotic complications and decreased during the resolution of these events. PMID- 11734750 TI - Hemolytic-uremic syndrome associated with Coxiella burnetii infection. AB - Acute Q fever most commonly manifests as a self-limited febrile illness, pneumonia or hepatitis. We report the case of a 12-year-old child with documented exposure to sources of Coxiella burnetii who was admitted to our hospital because of hemolytic-uremic syndrome. Serologic tests established the diagnosis of acute Q fever. PMID- 11734751 TI - Acute polyosteal osteomyelitis in a patient with congenital ichthyosis and acute lymphoblastic leukemia. AB - Osteomyelitis can develop after bacteremia in children and is a rare complication of chemotherapy for acute lymphoblastic leukemia. A child with congenital ichthyosis, acute lymphoblastic leukemia and multifocal hematogenous osteomyelitis is described herein. A breached skin barrier secondary to ichthyosis during induction chemotherapy, coupled with impaired immunity, likely provided the opportunity for bacterial seeding, leading to an extensive multifocal osteomyelitis. PMID- 11734752 TI - Facial palsy associated with mumps parotitis. AB - Peripheral facial nerve paralysis is relatively common in the pediatric age group. However, facial palsy rarely has been documented in patients with mumps parotitis. This case report describes the findings of a 3-year-old Japanese boy who developed facial palsy immediately after mumps parotitis. This work calls attention to a possible association between mumps parotitis and facial palsy. PMID- 11734753 TI - Apnea, bradycardia and thrombocytopenia in a premature infant. PMID- 11734754 TI - Antibiotic therapy of an enterococcal ventriculoperitoneal shunt infection. PMID- 11734755 TI - Dangers of empiric oral ciprofloxacin in the treatment of acute inflammatory diarrhea in children. PMID- 11734756 TI - Stevens-Johnson syndrome after vaccination. PMID- 11734757 TI - Interleukin-8 concentrations in the peripheral circulation of human immunodeficiency virus type 1-infected children suggest blunted chemokine responses. PMID- 11734759 TI - Bacteriologic and clinical efficacy of high dose amoxicillin/clavulanate in children with acute otitis media. AB - OBJECTIVES: To determine the bacteriologic and clinical efficacy of high dose amoxicillin/clavulanate (90/6.4 mg/kg/day) against common bacterial pathogens causing acute otitis media (AOM), including penicillin-resistant Streptococcus pneumoniae (PRSP). METHODS: In this open label multicenter study, 521 infants and children with AOM [mean age, 18.6 months; age < 24 months, n = 375 (72%)] were treated with amoxicillin/clavulanate 90/6.4 mg/kg/day in two divided doses for 10 days. Bilateral otitis media, previous episodes of AOM, antibiotic treatment within 3 months and day-care attendance were recorded in 60.1, 35.7, 50.2 and 38.2% of the children, respectively. Tympanocentesis was performed before the first dose and repeated on Days 4 to 6 for all children with S. pneumoniae at 22 centers and for all children with any pathogen at 3 centers. Clinical response was assessed at end of therapy. RESULTS: Pathogens were isolated from 355 (68%) of 521 enrolled children; 180 children underwent repeat tympanocentesis and were bacteriologically evaluable. Baseline pathogens were S. pneumoniae (n = 122 enrolled/93 bacteriologically evaluable), Haemophilus influenzae (n = 160/51), both (n = 37/32) and others (n = 36/4). Pathogens were eradicated from 172 (96%) of 180 bacteriologically evaluable children. Overall 122 (98%) of 125 isolates of S. pneumoniae were eradicated, including 31 (91%) of 34 PRSP isolates (penicillin MICs 2 to 4 micrograms/ml). Seventy-eight (94%) of 83 isolates of H. influenzae were eradicated. Symptoms and otoscopic signs of acute inflammation were completely resolved or improved on Days 12 to 15 in 263 (89%) of 295 clinically evaluable children with bacteriologically documented AOM. CONCLUSIONS: On the basis of bacteriologic outcome on Days 4 to 6 and clinical outcome on Days 12 to 15, we found that high dose amoxicillin/clavulanate (90/6.4 mg/kg/day) was highly efficacious in children with AOM, including those most likely to fail treatment, namely children < 24 months of age and those with infectious caused by PRSP. PMID- 11734760 TI - Pneumococcal meningitis in children: relationship of antibiotic resistance to clinical characteristics and outcomes. AB - BACKGROUND: The relationship of antibiotic susceptibility to clinical outcome in children with pneumococcal meningitis is uncertain. Previous studies have been limited by inclusion of relatively few patients infected with nonsusceptible pneumococci and inconsistent use of empiric vancomycin. METHODS: Medical records of 86 children with culture-confirmed pneumococcal meningitis at a single institution from October, 1991, to October, 1999, were retrospectively reviewed, and differences in presentation and outcome based on antibiotic susceptibility of pneumococcal isolates were assessed. RESULTS: Of 86 isolates 34 were nonsusceptible to penicillin (12 resistant). Of 60 isolates for which cefotaxime susceptibility data were available, 17 were nonsusceptible (12 resistant). Antibiotic susceptibility was not significantly associated with death, intensive care unit admission, mechanical ventilation, focal neurologic deficits, seizures, secondary fever, abnormal neuroimaging studies or hospital days. Children with penicillin-resistant isolates had significantly higher median blood leukocyte counts (24,100/microliter vs. 15,700/microliter, P = 0.03) and lower median CSF protein concentrations (85 mg/dl vs. 219 mg/dl, P = 0.04), were more likely to have a CSF glucose concentration of > or = 50 mg/dl (7 of 11 vs. 15 of 68, P = 0.009) and had lower rates of sensorineural hearing loss (1 of 8 vs. 25 of 40, P = 0.02) than children with isolates that were not resistant to penicillin. Children with cefotaxime-nonsusceptible isolates had an increased median duration of primary fever compared with those with nonsusceptible strains (6 days vs. 3.5 days, P = 0.02). CONCLUSIONS: In children with pneumococcal meningitis, penicillin resistance was associated with a reduced risk of hearing loss, while cefotaxime resistance was associated with a longer duration of fever. Other outcome measures were not significantly influenced by the antibiotic susceptibility of pneumococcal isolates. PMID- 11734761 TI - Pichia anomala outbreak in a nursery: exogenous source? AB - BACKGROUND: Pichia anomala is a rare cause of fungemia. From February to April, 1998, eight cases of fungemia occurred in the intensive care and high risk units of the Nursery. There were four infants with P. anomala infection, one of whom also had Candida parapsilosis infection, two cases with C. parapsilosis infection and two with Candida albicans infection. OBJECTIVE: To determine factors associated with fungemia in the intensive care and high risk units of the Nursery, especially P. anomala. METHODS: A cohort study with 59 newborns. RESULTS: Factors associated with fungemia were: central venous catheter (CVC) (P = 0.0006); total parenteral nutrition (TPN) (P = 0.0005); lipid emulsion (P = 0.002); previous antimicrobial use (P = 0.002); and other invasive procedures (P = 0.002). Factors associated with P. anomala fungemia were: CVC (P = 0.004); TPN (P = 0.018); previous antibiotic use (P = 0.037); and other invasive procedures (P = 0.037). Evaluation of the units demonstrated that there were several technical problems involving administration of TPN that was manipulated in the Nursery without precautions. Changes in TPN formulation and education as to adequate technique were implemented. During follow-up (1998 to 1999) only two fungemias occurred that were caused by C. albicans. Cultures of hands of personnel were negative for P. anomala. Electrophoretic karyotyping of P. anomala showed three profiles. CONCLUSIONS: Factors associated with fungemia were catheter use, invasive procedures and total parenteral nutrition, suggesting that the acquisition of P. anomala was exogenous. PMID- 11734762 TI - Clinical severity of Norwalk virus and Sapporo virus gastroenteritis in children in Hokkaido, Japan. AB - OBJECTIVE: To clarify the clinical significance and etiologic impact of Norwalk virus (NV) and Sapporo virus (SV) in viral gastroenteritis in Japanese children. STUDY DESIGN: Two outbreaks each of NV gastroenteritis and SV gastroenteritis occurring in an infant home in Sapporo, Japan, as well as 95 hospitalized children with acute gastroenteritis were retrospectively evaluated using a 0- to 20-point clinical severity scoring system. RESULTS: The mean severity scores for NV and SV gastroenteritis outbreaks were 7.9 and 5.2, respectively, as compared with 8.4 for rotavirus A gastroenteritis that occurred in the same infant home. Among 95 hospitalized children with acute gastroenteritis, rotavirus A was detected in 47% followed by NV in 18%. SV was not found. CONCLUSION: Our data indicate that NV can cause severe gastroenteritis and is an important etiologic agent in hospitalized cases, whereas SV causes mild gastroenteritis in Japanese children. PMID- 11734763 TI - Identification and quantification of ureaplasmas colonizing the respiratory tract and assessment of their role in the development of chronic lung disease in preterm infants. AB - BACKGROUND: The role of Ureaplasma urealyticum in the development of chronic lung disease (CLD) in preterm infants continues to be disputed. Recently U. urealyticum has been found to consist of two species, U. urealyticum and Ureaplasma parvum, a finding that has not been considered in previous studies of CLD. This study examined the possible relationships between development of CLD and respiratory colonization by these newly redefined species, their concentrations in lower respiratory secretions and the effect of pulmonary surfactant treatment on these relationships in preterm infants with birth weights < 1500 g. METHODS: Endotracheal aspirates (ETA) were collected from intubated infants when airway suctioning was medically required. ETA were stored at -80 degrees C until quantitative cultures for ureaplasmas and Mycoplasma hominis were performed. Culture results were correlated with development of CLD. RESULTS: Of 475 infants (birth weights < 1500 g) admitted during the 2-year study period, 272 were excluded because they were not intubated or were extubated before ETA could be obtained. An additional 28 infants died, were discharged or were transferred before they could be assessed for CLD. From the remaining 175 infants ureaplasmas were isolated from 66 (38%). No statistically significant associations were identified between development of CLD and the Ureaplasma species isolated, or concentration of ureaplasmas in lower respiratory secretions. These findings were not altered by treatment with pulmonary surfactant (Survanta). CONCLUSION: Lower respiratory colonization by ureaplasmas does not appear to be a contributory cause of CLD in preterm infants. PMID- 11734764 TI - Community-acquired methicillin-resistant Staphylococcus aureus infections in south Texas children. AB - BACKGROUND: Community-acquired methicillin-resistant Staphylococcus aureus (CAMRSA) infections have increased dramatically from 1990 to 2000. OBJECTIVES: The objectives of this retrospective study were to report the frequency of CAMRSA isolates, to describe the spectrum of disease observed in children infected with CAMRSA and to compare the antibiotic susceptibility patterns of community acquired and nosocomial methicillin-resistant S. aureus (MRSA) infections. METHODS: All cases of S. aureus including MRSA infections were identified by a computer-assisted search of the Vitek system culture results from 1990 to 2000 and review of the minutes of Infection Control Committee meetings. RESULTS: MRSA was isolated from 147 children (77 boys; 2 weeks to 17 years) from October 1, 1990, to December 31, 2000. Seven cases of CAMRSA were identified from 1990 to 1996, and 53 cases were identified from 1997 to 2000 with 35 cases occurring in 2000. Of the 128 children whose medical records were reviewed, 60 (47%) had CAMRSA infections. In 53 (88%) of these 60 children no risk factor for MRSA was identified. Soft tissue infections accounted for 48 (91%) of the 53 cases of the CAMRSA in children without known risk factors. CAMRSA isolates from children without known risk factors were more susceptible to trimethoprim-sulfamethoxazole (98% vs. 82%; P < 0.005) and clindamycin (92% vs. 57%; P < 0.001) and less susceptible to tetracycline (54% vs. 95%; P < 0.001) than were nosocomial MRSA isolates. CONCLUSIONS: The emergence of CAMRSA as a cause of common infections may require a change in the initial selection of antibiotics to assure appropriate coverage in critically ill children. PMID- 11734765 TI - High prevalence of erythromycin resistance of Streptococcus pyogenes in Greek children. AB - BACKGROUND: Macrolide resistance among Streptococcus pyogenes strains is increasing in many European countries. Greece was not considered a country with high prevalence of macrolide-resistant S. pyogenes strains, and until now the genetic mechanism of resistance was unknown. METHODS: During the 25-month period from December, 1998, to December, 2000, pharyngeal cultures for S. pyogenes were performed on 743 Greek children with the clinical diagnosis of pharyngitis. The children were 1 to 16 years old (median age, 7 years) and were living in Central and Southern Greece. S. pyogenes isolates were tested for their susceptibility to erythromycin, clarithromycin, azithromycin, clindamycin, penicillin G, amoxicillin/clavulanate and cefprozil. The erythromycin-resistant isolates were further studied for their genetic mechanism of resistance by means of PCR. RESULTS: Of a total of 275 S. pyogenes isolates recovered, 105 (38%) were erythromycin-resistant (MIC > or = 1 microgram/ml) [corrected], with 54, 45 and 1% of them carrying mef(A), erm(A) [subclass erm(TR)] and erm(B) gene, respectively. The prevalence of erythromycin-resistant strains was 29 and 42% during the time periods December, 1998, to December, 1999, and January, 2000, to December, 2000, respectively. All erythromycin-resistant isolates were also resistant to clarithromycin and azithromycin. The isolates carrying the erm(A) gene were inducibly resistant to clindamycin. The 275 S. pyogenes isolates had ceprozil MICs < or = 0.032 microgram/ml. CONCLUSIONS: The current high (38%) prevalence of erythromycin-resistant S. pyogenes in Central and Southern Greece requires continuous surveillance and careful antibiotic policy. PMID- 11734766 TI - Prospective study of 125 cases of Staphylococcus aureus bacteremia in children in New Zealand. AB - BACKGROUND: Staphylococcus aureus bacteremia is a common complication of S. aureus infection. There are few pediatric studies defining the incidence and associated morbidity and mortality of S. aureus bacteremia and no such New Zealand studies. We conducted a prospective study of S. aureus bacteremia in children in New Zealand. METHODS: From July 1, 1996 to December 31, 1998, we included all children < 16 years of age with S. aureus bacteremia in Auckland and Christchurch. Relevant information regarding patient demographics, clinical course and outcome and laboratory results was recorded. RESULTS: One hundred twenty-five cases of true S. aureus bacteremia were identified. There were 4 deaths within 30 days of the onset of bacteremia. Fourteen (11%) of the children were < 1 month of age. Maori children (relative risk, 2.0; 95% confidence interval, 1.3 to 3.2) were twice as likely and Pacific Island children (relative risk, 2.5; 95% confidence interval, 1.6 to 3.8) 2.5 times as likely as white children to acquire S. aureus bacteremia. The peak incidence of S. aureus bacteremia was observed in Pacific Island children < 1 year of age (105 cases/100,000 children/year). Twenty-seven percent of cases were related to intravenous catheters. Seventy percent of cases were community-acquired. Ninety eight percent of non-catheter-related cases in children > 1 month of age were community-acquired. There was a low rate of methicillin resistance (6%). CONCLUSIONS: S. aureus bacteremia is largely community-acquired in children in New Zealand and is more common in Pacific Island and Maori populations. Although there is a low associated mortality, a significant number are potentially preventable cases secondary to intravenous catheters. PMID- 11734767 TI - Hospitalization rates for respiratory syncytial virus infection in premature infants born during two consecutive seasons. AB - OBJECTIVE: To collect data on hospitalization rates for respiratory syncytial virus (RSV) illness during the season of 1999 to 2000 in nonprophylaxed premature infants < or = 32 weeks gestational age (GA) in Spain and compare this with previously published data collected in the season of 1998 to 1999. METHODS: Children born at < or = 32 weeks GA between April 1, 1999, and April 31, 2000, and discharged from the hospital before April 31, 2000, were included. Neonatal and demographic data were obtained at the initial visit. Study subjects were followed at monthly intervals throughout the respiratory season. RSV status and morbidity data were collected on patients rehospitalized for respiratory illness. RESULTS: The 999 evaluable patients in the 2000 season were comparable to the 1999 sample, except for higher rates of family allergy history and number of multiple deliveries and a lower rate of neonatal morbidity. The hospitalization rate for RSV illness was 13.4% in the 1999 season and 13.1% in the 2000 season; 10 (8%) were RSV reinfections in the 2000 season. Significant independent prognostic variables for high risk of RSV hospital admission included: lower gestational age; chronologic age < 3 months at onset of the RSV season; living with school age siblings; and exposure to tobacco smoke. CONCLUSIONS: Hospitalization rates for RSV disease in nonprophylaxed preterm infants < or = 32 weeks GA were high in Spain and comparable during two consecutive RSV seasons (13%). Readmission for a second RSV infection was also common. PMID- 11734768 TI - Population-based active surveillance for neonatal group B streptococcal infections in Alberta, Canada: implications for vaccine formulation. AB - BACKGROUND: Knowledge of circulating serotypes of group B Streptococcus (GBS) is important for formulation of vaccines. There are no Canadian data on the serotype distribution of neonatal GBS isolates. METHODS: Using a retrospective laboratory and health record survey between 1993 and 1994 (before introduction of Canadian prevention guidelines) and prospective active laboratory-based surveillance from 1995 to 1999 of all laboratories in Alberta, we identified 168 cases of invasive neonatal GBS infections including stillbirths among 262,398 total births; 118 of 123 (96%) isolates from 1995 to 1999 were serotyped, and the corresponding neonatal health records were reviewed. RESULTS: The average annual incidence was 0.64 of 1000 total births/year. Of these 95 (57%) had early onset disease (EOD), 15 (9%) were still births and 58 (34%) had late onset disease (LOD). Eighty-one percent of EOD cases were caused by serotypes Ia, Ia/c, Ia/c/R, III, III/R and V, V/R, whereas 81% of LOD cases were caused by serotypes III and III/R. GBS serotypes containing the C protein along with serotypes III and V as a group constituted 91% (107 of 118) of all GBS cases in our population. The most common clinical presentation was bacteremia without focus (74%) followed by meningitis (14%) and pneumonia (12%). During 1995 to 1999, in addition to 13 stillbirths, there were 6 of 64 (9%) neonatal deaths among EOD cases and 1 of 46 (2%) neonatal death among LOD cases. CONCLUSIONS: In this population-based study stillbirths account for a proportion of cases that are not routinely counted and represent a group for which intrapartum antibiotics would likely not be effective, but potentially preventable by vaccination. Inclusion of serotypes Ia, III and V in a conjugate vaccine or serotypes III and V conjugated with the C protein in a GBS vaccine could theoretically provide protection against the majority of GBS invasive disease in Alberta neonates. PMID- 11734769 TI - Bactericidal/permeability-increasing protein: clinical development and potential pediatric applications. PMID- 11734770 TI - Vaccines and autism. PMID- 11734771 TI - Granulocyte function disorders: aspects of development, genetics and management. AB - The field of phagocytic disorders has attained major biologic and clinical significance in the past 40 years. The development of exciting new techniques in molecular biology and the cellular physiology of signal transduction have made it possible to identify the genetic defects involved in many of these disorders. Moreover through immunopharmacologic intervention, bone marrow or peripheral or cord blood stem cell transplantation along with the prospect of gene therapy, we have begun attempts to at least partially correct genetic defects in cell development and activation pathways in the entire spectrum of phagocyte disorders. Carrier detection and prenatal diagnosis employing with chain reaction techniques or direct nucleotide sequencing in fetal blood have made these diseases potentially preventable or treatable in utero or shortly after birth. PMID- 11734772 TI - Abdominal actinomycosis with hydronephrosis in childhood. AB - Two 11-year-old boys had similar clinical courses of abdominal actinomycosis complicating ruptured appendicitis and manifesting as abdominal masses causing hydronephrosis. Cure was effected by surgery and prolonged penicillin therapy. PMID- 11734773 TI - Spinal intramedullary cysticercosis in a five-year-old child: case report and review of the literature. AB - Spinal intramedullary cysticercosis is rare and usually afflicts adults. We report the case of a 5-year-old Mexican girl with back pain who had a complex thoracic spinal intramedullary mass on magnetic resonance imaging and a positive immunoblot for Taenia solium. Surgery revealed a cystic mass containing a cysticercus. Cysticercosis should be suspected as the cause of an intramedullary spinal mass in a patient from an endemic area. PMID- 11734774 TI - Isolated cerebellar edema and obstructive hydrocephalus in a child with cerebral malaria. AB - An important complication of cerebral malaria is increased intracranial pressure which, when severe, is associated with high mortality or neurologic sequelae. We describe a 7-month-old child with cerebral malaria for whom cerebellar edema and obstructive hydrocephalus were initial radiologic findings. Despite significant hydrocephalus, the child had normal intracranial pressure during the course of the infection, and he recovered with minimal sequelae. PMID- 11734775 TI - Lack of pneumococcal antigenuria in infants immunized with heptavalent pneumococcal conjugate vaccine. AB - The incidence and duration of pneumococcal antigenuria were determined in 21 infants who received the first dose of pneumococcal conjugate vaccine. Urine specimens collected from the study participants at 1, 3, 6 and 9 days after immunization did not reveal the presence of pneumococcal antigen. PMID- 11734776 TI - Parotid abscess caused by Mycobacterium tuberculosis. AB - Tuberculosis of the parotid gland is rare. A 16-month-old US-born male infant with immigrant parents from Sudan presented to his primary care physician with periorbital cellulitis and preauricular lymphadenitis. He underwent incision and drainage of an abscess in the right intraparotid lymph node. The aspirate was positive for acid-fast bacilli by auramine-rhodamine stain and subsequently grew Mycobacterium tuberculosis. Antitubercular medications were started postoperatively. PMID- 11734777 TI - Fatal sepsis associated with acute pancreatitis caused by Moraxella catarrhalis in a child. AB - We describe a 4-year-old boy with Cornelia de Lange syndrome who died of septic shock caused by Moraxella catarrhalis bacteremia. At autopsy there was evidence of acute hemorrhagic pancreatitis with abscesses. Gram-negative diplococci were seen histologically in the abscesses and pancreatic ducts. PMID- 11734778 TI - Herpes zoster during varicella. AB - A 4-year-old boy had varicella infection. Two days later vesicular lesions clustered within the left 10th thoracic dermatome. Varicella-zoster virus IgM antibody in serum was positive. He was diagnosed with varicella infection combined with herpes zoster. This is the first case report in the medical literature. PMID- 11734779 TI - Bacteremia with group A Streptococcus associated with herpetic gingivostomatitis. PMID- 11734780 TI - Tissue Doppler imaging: a useful echocardiographic method for the cardiac sonographer to assess systolic and diastolic ventricular function. AB - Tissue Doppler imaging (TDI) has evolved to become a useful noninvasive method that can complement other echocardiographic techniques in the assessment of left ventricular myocardial velocities in a variety of clinical conditions. Color 2 dimensional and color M-mode TDI are used for quantification of systolic myocardial velocities during myocardial ischemia at rest and with pharmacologic stress testing. Spectral pulsed TDI can provide measurements of regional systolic and diastolic myocardial velocities, and it is particularly useful in the identification of abnormalities of left ventricular diastolic relaxation. This review summarizes the clinical applications of TDI to promote understanding of its utility in the evaluation of left ventricular myocardial function. PMID- 11734781 TI - Comparison of myocardial tissue Doppler with transmitral flow Doppler in left ventricular hypertrophy. AB - We sought to determine the most useful echocardiographic measurements for assessment of diastolic function in patients with left ventricular hypertrophy (LVH) and normal systolic function. We compared myocardial Doppler velocities of the basal inferoposterior wall with mitral inflow pulsed wave Doppler velocities in 11 healthy volunteers (age, 36 +/- 6 years), 25 patients (age, 64 +/- 14 years) without LVH, and 37 patients (age, 67 +/- 14 years) with LVH and otherwise normal echocardiograms. The discriminatory measurements were myocardial A-wave duration (120 +/- 18 versus 98 +/- 20 and 92 +/- 12 ms, P <.0001), myocardial isovolumetric relaxation time (124 +/- 45 versus 95 +/- 48 and 78 +/- 25 ms, P =.0035), mitral A-wave velocity (0.98 +/- 0.37 versus 0.73 +/- 0.28 m/s and 0.61 +/- 0.22 m/s, P =.009), and mitral E-wave deceleration time (257 +/- 93 versus 201 +/- 85 ms and 184 +/- 83 ms, P =.015), which were significantly increased, and myocardial E-wave velocity (0.84 +/- 0.04 m/s versus 0.13 +/- 0.03 m/s and 0.14 +/- 0.03 m/s, P <.0001), which was significantly decreased, in patients with LVH compared with patients without LVH and normal volunteers, respectively. Left ventricular posterior wall thickness correlated with myocardial isovolumetric relaxation time (r = 0.52, P <.0001) and myocardial A-wave duration (r = 0.59, P <.0001), negatively with myocardial E wave (r = -0.43, P <.0001), and showed no correlation with mitral inflow parameters except mitral inflow A wave (r = 0.43, P =.002). On multivariate analysis using these variables, myocardial isovolumetric relaxation time (P =.0014) and A-wave duration (P =.001) were the only 2 variables that correlated with posterior wall thickness (multiple R = 0.71). In the presence of LVH and preserved left ventricular systolic function, myocardial relaxation time and velocities are more sensitive than mitral Doppler inflow parameters in detecting abnormal left ventricular relaxation. PMID- 11734782 TI - Effects of acute decrease in systemic afterload on accuracy of Doppler-derived left ventricular rate of pressure rise measurement in anesthetized patients. AB - OBJECTIVES: This study was designed to describe exactly the effects of acute decrease in systemic afterload on the accuracy of Doppler-derived left ventricular rate of pressure rise (LV DeltaP/Delta(t)) measurements compared with other routinely used indices of systolic function. METHODS: Twelve patients scheduled for coronary artery bypass grafting were studied. After induction of anesthesia (T0), afterload was modified by incremental administrations of nicardipine (T1-4). At each step of the procedure, thermodilution-derived cardiac index, left ventricular (LV) fractional area change, and LV DeltaP/Delta(t) were measured, and systemic vascular resistances were calculated. RESULTS: During the procedure, the systemic vascular resistances decrease averaged 13.4%. Systemic vascular resistances were correlated with LV DeltaP/Delta(t) (r = 0.843, P =.003) but inversely correlated with cardiac index (r = -0.782, P =.005) and LV fractional area change (r = -0.887, P =.003). CONCLUSION: In conclusion, and inversely to cardiac index or LV fractional area change, LV DeltaP/Delta(t) does not overestimate LV contractility in the presence of an acute decrease in systemic afterload. PMID- 11734783 TI - Echocardiographic characterization of fundamental mechanisms of abnormal diastolic filling in diabetic rats with a parameterized diastolic filling formalism. AB - Abnormalities of diastolic function (DF) precede systolic dysfunction in diabetic cardiomyopathy. Transmitral Doppler flow analysis is the primary method for noninvasively assessing DF. We used model-based Doppler E-wave analysis to evaluate diastolic function differences between normal and diabetic rat hearts. Control rats and those with diabetes underwent echocardiography with analysis by traditional Doppler indexes and by the parameterized diastolic filling (PDF) formalism, generating 3 parameters, x0, c, and k, that uniquely characterize each E-wave. Significant intergroup differences in the E/A ratios (P <.01), isovolumic relaxation times (P <.01), and the modeling parameter c (P <.05) were found. There were no significant differences in shortening fraction, deceleration time, myocardial collagen content, or the parameters x0 and k between diabetic and control rats. These results indicate that differences in diastolic function may be noninvasively quantified and that diabetic hearts may exhibit defects in uncoupling of the contractile apparatus without concomitant increases in chamber stiffness. PMID- 11734784 TI - Quantification of myocardial perfusion and determination of coronary stenosis severity during hyperemia using real-time myocardial contrast echocardiography. AB - Although regional myocardial perfusion can be currently quantified with myocardial contrast echocardiography (MCE) by using intermittent harmonic imaging (IHI), the method is tedious and time-consuming in the clinical setting. We hypothesized that regional myocardial perfusion can be quantified and the severity of coronary stenosis determined during hyperemia with MCE using real time imaging (RTI) where microbubbles are not destroyed. Six open-chest dogs were studied during maximal hyperemia induced by adenosine in the absence or presence of coronary stenoses varying from mild to severe. Myocardial blood flow (MBF) was measured at each stage by using radiolabeled microspheres. MCE was performed using both IHI and RTI. Data for the latter were acquired in both end-systole and end-diastole. No differences were found between myocardial flow velocity (MFV) derived from IHI and RTI when end-systolic frames were used for the latter. MFV was consistently higher for RTI (P <.01) when end-diastolic frames were used. A linear relation was noted between MFV and radiolabeled microsphere-derived MBF ratios from the stenosed and the normal beds when end-systolic frames were used for RTI (r = 0.78, P <.001), whereas no relation was found when end-diastolic frames were used (r = 0.08, P =.78). The scatter for assessing MBF (A.beta) was minimal for IHI and RTI (9%-10%) with end-systolic frames, whereas that for RTI with end-diastolic frames was large (30%). Furthermore the correlation with radiolabeled microsphere-derived MBF was significantly (P <.01) weaker with RTI when end-diastolic frames were used (r = 0.53) than when end-systolic frames (r = 0.94) or IHI was used (r = 0.99). Data acquisition for IHI was 10 minutes, whereas it was 8 seconds for RTI. Thus, RTI can be used to quantify regional myocardial perfusion and stenosis severity during MCE. Only end-systolic frames, however, provide accurate data. RTI offers a rapid and easy means of assessing regional myocardial perfusion with MCE. PMID- 11734785 TI - Contrast-enhanced quantitation of left ventricular ejection fraction: what is the best method? AB - The need to enhance the echocardiographic determination of left ventricular ejection fraction (LVEF) is greatest in patients with suboptimal images. We have previously demonstrated that in difficult-to-image patients, contrast-enhanced power harmonic imaging resulted in accurate calculation of LVEF by using Simpson's method. However, the incremental accuracy of contrast enhancement with other methods of determining LVEF has not been examined. This study prospectively assessed the comparative accuracy of LVEF determination by using the Quinones' method with fundamental imaging (FU), tissue harmonic imaging (TH), contrast enhanced harmonic imaging [TH(CON)], as well as Quinones' method with contrast enhanced power harmonic imaging [POW(QUIN)] in 62 patients with suboptimal images. LVEF was also calculated by using contrast-enhanced power harmonic imaging and Simpson's method [POW(SIMP)] in these patients. We demonstrated that LVEF calculated from the POW(QUIN) mode had the best agreement with radionuclide angiography (standard of comparison) compared with FU, TH, and TH(CON). However, POW(SIMP) was even more accurate. In conclusion, when the Quinones' method was used to calculate LVEF in difficult-to-image patients, POW(QUIN) mode was the most accurate. However, POW(SIMP) was even more accurate and should be the method of choice when a high degree of quantitative accuracy is required. PMID- 11734786 TI - An assessment of tissue harmonic versus fundamental imaging modes for echocardiographic measurements. AB - The study aimed to evaluate whether a significant difference exists between tissue harmonic imaging (THI) and fundamental imaging (FI) in routine quantitative echocardiographic assessment. We also examined the effects of THI on endocardial definition (ED). Fifty-eight unselected patients (29 men, 29 women; mean age 53 years) referred for transthoracic echocardiography were studied with use of both FI and THI modes. Two independent observers made M-mode measurements of the following: left atrium, aortic root, and left ventricular internal dimensions and wall thickness; they also measured left ventricular outflow tract diameter and left atrial area from 2-dimensional images. Endocardial definition was assessed with use of an arbitrary scoring system (0 = endocardium not visualized, 1 = endocardium visualized but suboptimally, 2 = endocardium well defined) and the American Society of Echocardiography 16-segment model. No significant difference existed between cardiac measurements derived from FI and THI. However, a highly significant improvement in ED occurred with THI (ED index 1.83 versus 1.70, P <.0001). This study suggests that no systematic differences exist in standard echocardiographic measurements between THI and FI, even in the setting of improved ED. PMID- 11734787 TI - Use of peak Doppler gradient across ventricular septal defects leads to underestimation of right-sided pressures in patients with "sloped" Doppler signals. AB - In patients with "sloped" appearance of the Doppler signal across a ventricular septal defect (VSD), the peak Doppler velocity seems to overestimate the catheterization-derived peak-to-peak gradient, resulting in underestimation of right-sided heart pressures. In 11 patients with sloped Doppler signals across the VSD, ventricular pressure tracings were compared with simultaneous recordings of the Doppler signal. The average peak Doppler gradient (40.2 +/- 19.2 mm Hg) overestimated the catheterization-derived peak-to-peak gradient (20.2 +/- 13.6 mm Hg) significantly (P < or =.001). Doppler mean gradient (20.2 +/- 11.3 mm Hg; P = ns) and end-systolic gradient (17.0 +/- 12.5 mm Hg; P < or =.05) were closer estimates of the catheterization peak-to-peak gradient. All Doppler gradients showed good correlation to the catheterization peak-to-peak gradient with r2 values of 0.77, 0.73, and 0.91. We conclude that Doppler mean or end-systolic gradients should be used for calculation of right-sided heart pressures in this patient population. PMID- 11734788 TI - Quantification of extracranial carotid artery stenosis by ultrafast three dimensional ultrasound. AB - This study was designed to evaluate the possible contribution of 3-dimensional (3D) ultrasound (US) for noninvasive detection of extracranial carotid artery stenosis. Sixty-nine stenotic lesions of extracranial carotid artery were studied by (1) B-mode (Bm) US, (2) Doppler spectral analysis, and (3) a prototype of 3D vascular system. When indicated (46 stenotic lesions), biplane carotid angiography (CA) was performed. The degree of luminal narrowing measured as percent area reduction in the 3D data set correlated well with the degree of stenosis estimated by CA (r = 0.79, P <.01, mean difference 7.8% +/- 15.5%); however, for stenosis between 40% and 70%, 3D US tended to give higher values. Compared with CA, the sensitivity, specificity, diagnostic accuracy, and positive predictive value of 3D US for significant (> or =70%) stenosis were 96.0%, 77.7%, 88.3%, and 85.7%, respectively. Thus, 3D US showed good sensitivity and diagnostic accuracy for detection of significant stenosis of extracranial carotid artery. For stenosis between 40% and 70%, 3D US indicated a higher degree than CA; this finding suggests that CA may underestimate the severity of stenosis due to known discrepancies between linear measurement and true anatomic situation. PMID- 11734789 TI - Licensure of the cardiac sonographer: an overview of issues and activities. AB - The impact of echocardiography on the continuum of cardiovascular health care is well established. Ongoing concerns regarding costs, accessibility, quality, and appropriateness of services rendered by practitioners of echocardiography have prompted various legislative proposals and regulatory policies from government, medical professional groups, and health plans. Specifically, there continues to be a drive to enact law for licensure of sonographers. These activities require continuing advocacy for the profession with active leadership. As part of its mission statement, the American Society of Echocardiography (ASE) states, "ASE strives to be a leader in public policy in order to create a favorable environment for excellence in the practice of echocardiography." As such, the ASE is committed to an increase in their interaction with legislators, payers, and policy makers. This article describes the historical perspective of state, federal, and provincial sonographer licensure issues to provide an understanding of the political perspectives. PMID- 11734790 TI - Localized pericardial tamponade: difficult echocardiographic diagnosis of a rare complication after cardiac surgery. AB - We report 2 cases of localized pericardial tamponade occurring soon after cardiac surgery, in which the diagnosis could not be made with transthoracic echocardiography. Computed tomography and transesophageal echocardiography, respectively, were necessary, and this underlies the importance of alternative imaging modalities when this condition is suspected. A high index of suspicion is crucial for reaching the correct diagnosis. PMID- 11734791 TI - Eustachian valve cyst. AB - Transesophageal echocardiography provides high resolution of intracardiac structures. One potential problem is the misdiagnosis of previously unrecognized benign cardiac structures as malignant processes, leading to unnecessary surgical procedures. We describe an unusual case of benign intracardiac eustachian valve cyst monitored over 2 years. PMID- 11734792 TI - Acute myocardial infarction caused by paradoxical coronary embolization in a patient with a patent foramen ovale. AB - We describe the case of a young woman without cardiac risk factors who had an acute inferoapical myocardial infarction. Coronary angiographic appearance was consistent with thrombus in the distal left anterior descending coronary artery. A patent foramen ovale with moderate right-to-left shunting after the Valsalva maneuver was detected by contrast transesophageal echocardiography. No other cardioembolic source was identified. Paradoxical embolization through a patent foramen ovale is a rare phenomenon, which appeared to have resulted in myocardial infarction in this patient. PMID- 11734793 TI - Thrombolytic therapy of a left-sided prosthetic valve thrombosis without hemodynamic obstruction: a case report. AB - We report a case of a 38-year-old woman with a prosthetic mitral valve who presented with multiple embolic events. Transesophageal echocardiography was used to diagnose nonobstructive thrombi on the prosthetic valve. She underwent successful thrombolytic therapy. The patient was discovered to be in a hypercoagulable state, which probably was caused by the concomitant use of phenytoin. We review the literature for diagnosis and treatment of nonobstructive prosthetic valve thrombosis. PMID- 11734796 TI - Infrainguinal vein graft surveillance: the criteria for intervention. AB - In most patients, infrainguinal vein grafts are commonly performed for limb threatening ischemia. Once limb salvage has been achieved, most clinicians agree that regular appointments with the vascular team are important to monitor graft patency. However, the timing of these appointments and what defines an adequate examination remains debatable. The following cases illustrate the natural history and interventions performed in 2 such patients. A review of the literature and a commentary on surveillance strategies is provided to elucidate the advantages and disadvantages of currently available vascular laboratory testing. Recommendations also are made for the appropriate timing of intervention to revise these grafts. PMID- 11734795 TI - Patient outcomes after peripheral revascularization surgery. AB - Acquired peripheral vascular disease (PVD) is a significant problem in the United States, resulting in both morbidity and mortality. The purpose of the pilot study was to determine patient outcomes after peripheral revascularization surgery. The specific aims of the pilot study were to examine peripheral revascularization surgical patient outcomes (PVD-related clinical symptoms, functioning, atherosclerotic disease risk factor reduction, and patient satisfaction) and to determine the influence of selected patient demographic characteristics (gender, age) on selected patient outcomes (PVD-related clinical symptoms, functioning, atherosclerotic disease risk factor reduction, and patient satisfaction). A prospective, repeated measures design was used for the study. A total of 39 patients, 18 women and 21 men, participated in the study, with a mean age of 68.86 years (SD = 13.61). The average length of hospitalization after surgery was 4.05 days. At 1 month after discharge, the majority of patients had relief from claudication and paresthesia. In regard to outcomes related to atherosclerotic risk factor modification, patients reported that they exercised on a routine basis, an average of 5.31 +/- 1.97 times per week. Before surgery, 21 patients reported that they smoked; 6 patients reported that they continued to smoke at follow-up. There were no significant differences in mean total scores of atherosclerotic risk modification by either gender or age groups (<65 or > or =65 years) with the use of one-way analyses of variance (ANOVAs). By using a Likert scale (ie, 0 to 10), the mean level of functioning was 8.18 +/- 2.76, with women having significantly higher mean levels of functioning (F = 4.26, P <.05). Comparing baseline scores of functioning on the Medical Outcomes Study Short-Form 36 (MOS SF-36), there was a significant improvement (F = 2.11, P <.05) in general health subscale scores at 1 month after surgery. Subjects' mean overall satisfaction with the results of surgery, with a 0 to 10 scale, was 7.33 +/- 2.84. Again, by using one-way ANOVAs, females had significantly higher mean satisfaction rating than males (F = 4.52, P <.05). Although findings from this pilot study are limited in their generalizability, clinicians need to continue to evaluate opportunities to further reduce variability in clinical practice patterns for optimal patient outcomes. Study findings also indicated that additional interventions are warranted to educate and provide rehabilitation for patients regarding an exercise program and overall behavior modification strategies to reduce risk for atherosclerotic disease. PMID- 11734797 TI - Endovascular repair of a giant arteriovenous fistula: a case study. PMID- 11734799 TI - Geriatric best practices in nursing care. PMID- 11734798 TI - Establishing and running an effective telephone-based anticoagulation service. AB - Observational studies and randomized controlled trials have revealed improvement in international normalized ratio (INR) control and reduced thrombotic and hemorrhagic events in patients taking warfarin who are managed by an anticoagulation service (ACS) compared with traditional physician care. In this article, we describe how to establish a multidisciplinary telephone-based ACS to monitor INRs, dose warfarin, and heparin therapy, and to educate patients by telephone. We address how to improve ACS efficiency by using an electronic medical record, charting by exception, holding group-based education, communicating by telephone, and conducting quality assurance. We also make recommendations for improving the quality of care of patients taking anticoagulants that can be implemented in any setting and we discuss how to apply these guidelines to other remote disease-state management programs (eg, diabetes). PMID- 11734800 TI - Reality research: project time management and recruitment. PMID- 11734801 TI - Thinking like a coach. PMID- 11734802 TI - TIPS. PMID- 11734806 TI - The good, the bad and the ugly. PMID- 11734805 TI - Fallout from EuroWars. PMID- 11734807 TI - Proposed budget cuts threaten to short-circuit Grid network. PMID- 11734808 TI - Concern raised for missing biologist. PMID- 11734810 TI - Cores set to unearth hole picture of evolution. PMID- 11734811 TI - China caught out as model shows net fall in fish. PMID- 11734812 TI - First human clones get a cool response. PMID- 11734814 TI - Election result leaves Australian scientists fearful over funding. PMID- 11734813 TI - Protests as terror law targets foreigners at universities. PMID- 11734816 TI - The crystal ball of chaos. PMID- 11734817 TI - Goliath befriends David. PMID- 11734818 TI - An eye on the future. PMID- 11734819 TI - Japan's funding cuts hit the future of science. PMID- 11734820 TI - Could sale of fossils be the key to ending theft? PMID- 11734821 TI - Messenger RNA: origins of a discovery. PMID- 11734829 TI - Genetics of identity. PMID- 11734830 TI - When one whale matters. PMID- 11734831 TI - Physics at the attosecond frontier. PMID- 11734832 TI - Sea snow microcosms. PMID- 11734834 TI - Six steps to destruction. PMID- 11734835 TI - A new spin on black-hole masses. PMID- 11734836 TI - Accessory to kidney disease. PMID- 11734837 TI - A broader view of membranes. PMID- 11734839 TI - Asymmetric Broca's area in great apes. AB - Brodmann's area 44 delineates part of Broca's area within the inferior frontal gyrus of the human brain and is a critical region for speech production, being larger in the left hemisphere than in the right - an asymmetry that has been correlated with language dominance. Here we show that there is a similar asymmetry in this area, also with left-hemisphere dominance, in three great ape species (Pan troglodytes, Pan paniscus and Gorilla gorilla). Our findings suggest that the neuroanatomical substrates for left-hemisphere dominance in speech production were evident at least five million years ago and are not unique to hominid evolution. PMID- 11734840 TI - Immune defence in bumble-bee offspring. AB - Immune-challenged vertebrate females transfer specific antibodies to their offspring, but this gratuitous immunity cannot operate in invertebrates. Here we show that constitutive immune defence is enhanced in sexual offspring of the bumble-bee Bombus terrestris L. when the parental colony is immune-challenged. Our findings indicate that invertebrates may use a different component of the immune system to generate a facultative trans-generational increase in the immune response. PMID- 11734841 TI - Synthesis of carbon 'onions' in water. AB - The fabrication of carbon nanomaterials usually calls for expensive vacuum systems to generate plasmas and yields are disappointingly low. Here we describe a simple method for producing high-quality spherical carbon nano-'onions' in large quantities without the use of vacuum equipment. The nanoparticles, which have C60 cores surrounded by onion-like nested particles, are generated by an arc discharge between two graphite electrodes submerged in water. This technique is economical and environmentally benign, and produces uncontaminated nanoparticles which may be useful in many applications. PMID- 11734844 TI - Retraction: Furtive mating in female chimpanzees. PMID- 11734842 TI - Fossils and avian evolution. AB - Discoveries of archaic Mesozoic fossil birds ('opposite' birds, or enantiornithines) during the past decade have revolutionized our understanding of early avian evolution, but the rarity of Early Cretaceous ornithurines - birds that are closely related to the modern avian radiation - has meant that information about these species has lagged behind. PMID- 11734845 TI - Attosecond metrology. AB - The generation of ultrashort pulses is a key to exploring the dynamic behaviour of matter on ever-shorter timescales. Recent developments have pushed the duration of laser pulses close to its natural limit-the wave cycle, which lasts somewhat longer than one femtosecond (1 fs = 10-15 s) in the visible spectral range. Time-resolved measurements with these pulses are able to trace dynamics of molecular structure, but fail to capture electronic processes occurring on an attosecond (1 as = 10-18 s) timescale. Here we trace electronic dynamics with a time resolution of 5-7 M middle dot in circle challenge the conventional picture of black-hole formation in binary systems. Based on the mass estimate, we interpret the distinct X-ray variability of GRS1915+105 as arising from instabilities in an accretion disk that is dominated by radiation pressure, and radiating near the Eddington limit (the point where radiation pressure supports matter against gravity). Also, the mass estimate constrains most models which relate observable X-ray properties to the spin of black holes in microquasars. PMID- 11734848 TI - Atomic structure holography using thermal neutrons. AB - The idea of atomic-resolution holography has its roots in the X-ray work of Bragg and in Gabor's electron interference microscope. Gabor's lensless microscope was not realized in his time, but over the past twelve years there has been a steady increase in the number of reports on atomic-resolution holography. All of this work involves the use of electrons or hard X-rays to produce the hologram. Neutrons are often unique among scattering probes in their interaction with materials: for example, the relative visibility of hydrogen and its isotopes is a great advantage in the study of polymers and biologically relevant materials. Recent work proposed that atomic-resolution holography could be achieved with thermal neutrons. Here we use monochromatic thermal neutrons, adopting the inside source concept of Szoke, to image planes of oxygen atoms located above and below a single hydrogen atom in the oxide mineral simpsonite. PMID- 11734849 TI - Pressure-induced amorphization and an amorphous-amorphous transition in densified porous silicon. AB - Crystalline and amorphous forms of silicon are the principal materials used for solid-state electronics and photovoltaics technologies. Silicon is therefore a well-studied material, although new structures and properties are still being discovered. Compression of bulk silicon, which is tetrahedrally coordinated at atmospheric pressure, results in a transition to octahedrally coordinated metallic phases. In compressed nanocrystalline Si particles, the initial diamond structure persists to higher pressure than for bulk material, before transforming to high-density crystals. Here we report compression experiments on films of porous Si, which contains nanometre-sized domains of diamond-structured material. At pressures larger than 10 GPa we observed pressure-induced amorphization. Furthermore, we find from Raman spectroscopy measurements that the high-density amorphous form obtained by this process transforms to low-density amorphous silicon upon decompression. This amorphous-amorphous transition is remarkably similar to that reported previously for water, which suggests an underlying transition between a high-density and a low-density liquid phase in supercooled Si (refs 10, 14, 15). The Si melting temperature decreases with increasing pressure, and the crystalline semiconductor melts to a metallic liquid with average coordination approximately 5 (ref. 16). PMID- 11734850 TI - Real-time spectroscopy of transition states in bacteriorhodopsin during retinal isomerization. AB - Real-time investigations of the rearrangement of bonds during chemical transformations require femtosecond temporal resolution, so that the atomic vibrations within the reacting molecules can be observed. Following the development of lasers capable of emitting ultrashort laser flashes on this timescale, chemical reactions involving relatively simple molecules have been monitored in detail, revealing the transient existence of intermediate species as reactants are transformed into products. Here we report the direct observation of nuclear motion in a complex biological system, the retinal chromophore of bacteriorhodopsin (bR568), as it undergoes the trans-cis photoisomerization that is fundamental to the vision process. By using visible-light pulses of less than 5 femtosecond in duration, we are able to monitor changes in the vibrational spectra of the transition state and thus show that despite photoexcitation of the anti-bonding molecular orbital involved, isomerization does not occur instantly, but involves transient formation of a so-called 'tumbling state'. Our observations thus agree with growing experimental and ab initio evidence for a three-state photoisomerization model and firmly discount the initially suggested two-state model for this process. PMID- 11734851 TI - Systematic distortions in world fisheries catch trends. AB - Over 75% of the world marine fisheries catch (over 80 million tonnes per year) is sold on international markets, in contrast to other food commodities (such as rice). At present, only one institution, the Food and Agriculture Organization of the United Nations (FAO) maintains global fisheries statistics. As an intergovernmental organization, however, FAO must generally rely on the statistics provided by member countries, even if it is doubtful that these correspond to reality. Here we show that misreporting by countries with large fisheries, combined with the large and widely fluctuating catch of species such as the Peruvian anchoveta, can cause globally spurious trends. Such trends influence unwise investment decisions by firms in the fishing sector and by banks, and prevent the effective management of international fisheries. PMID- 11734852 TI - Demography of the endangered North Atlantic right whale. AB - Northern right whales (Eubalaena glacialis) were formerly abundant in the northwestern Atlantic, but by 1900 they had been hunted to near extinction. After the end of commercial whaling the population was thought to be recovering slowly; however, evidence indicates that it has been declining since about 1990 (ref. 1). There are now fewer than 300 individuals, and the species may already be functionally extinct owing to demographic stochasticity or the difficulty of females locating mates in the vast Atlantic Ocean (Allee effect). Using a data set containing over 10,000 sightings of photographically identified individuals we estimated trends in right whale demographic parameters since 1980. Here we construct, using these estimates, matrix population models allowing us to analyse the causes of right whale imperilment. Mortality has increased, especially among mother whales, causing declines in population growth rate, life expectancy and the mean lifetime number of reproductive events between the period 1980-1995. Increased mortality of mother whales can explain the declining population size, suggesting that the population is not doomed to extinction as a result of the Allee effect. An analysis of extinction time shows that demographic stochasticity has only a small effect, but preventing the deaths of only two female right whales per year would increase the population growth rate to replacement level. PMID- 11734853 TI - Transgenic DNA introgressed into traditional maize landraces in Oaxaca, Mexico. AB - Concerns have been raised about the potential effects of transgenic introductions on the genetic diversity of crop landraces and wild relatives in areas of crop origin and diversification, as this diversity is considered essential for global food security. Direct effects on non-target species, and the possibility of unintentionally transferring traits of ecological relevance onto landraces and wild relatives have also been sources of concern. The degree of genetic connectivity between industrial crops and their progenitors in landraces and wild relatives is a principal determinant of the evolutionary history of crops and agroecosystems throughout the world. Recent introductions of transgenic DNA constructs into agricultural fields provide unique markers to measure such connectivity. For these reasons, the detection of transgenic DNA in crop landraces is of critical importance. Here we report the presence of introgressed transgenic DNA constructs in native maize landraces grown in remote mountains in Oaxaca, Mexico, part of the Mesoamerican centre of origin and diversification of this crop. PMID- 11734854 TI - Kranz anatomy is not essential for terrestrial C4 plant photosynthesis. AB - An important adaptation to CO2-limited photosynthesis in cyanobacteria, algae and some plants was development of CO2-concentrating mechanisms (CCM). Evolution of a CCM occurred many times in flowering plants, beginning at least 15-20 million years ago, in response to atmospheric CO2 reduction, climate change, geological trends, and evolutionary diversification of species. In plants, this is achieved through a biochemical inorganic carbon pump called C4 photosynthesis, discovered 35 years ago. C4 photosynthesis is advantageous when limitations on carbon acquisition are imposed by high temperature, drought and saline conditions. It has been thought that a specialized leaf anatomy, composed of two, distinctive photosynthetic cell types (Kranz anatomy), is required for C4 photosynthesis. We provide evidence that C4 photosynthesis can function within a single photosynthetic cell in terrestrial plants. Borszczowia aralocaspica (Chenopodiaceae) has the photosynthetic features of C4 plants, yet lacks Kranz anatomy. This species accomplishes C4 photosynthesis through spatial compartmentation of photosynthetic enzymes, and by separation of two types of chloroplasts and other organelles in distinct positions within the chlorenchyma cell cytoplasm. PMID- 11734855 TI - Interactive memory systems in the human brain. AB - Learning and memory in humans rely upon several memory systems, which appear to have dissociable brain substrates. A fundamental question concerns whether, and how, these memory systems interact. Here we show using functional magnetic resonance imaging (FMRI) that these memory systems may compete with each other during classification learning in humans. The medial temporal lobe and basal ganglia were differently engaged across subjects during classification learning depending upon whether the task emphasized declarative or nondeclarative memory, even when the to-be-learned material and the level of performance did not differ. Consistent with competition between memory systems suggested by animal studies and neuroimaging, activity in these regions was negatively correlated across individuals. Further examination of classification learning using event-related FMRI showed rapid modulation of activity in these regions at the beginning of learning, suggesting that subjects relied upon the medial temporal lobe early in learning. However, this dependence rapidly declined with training, as predicted by previous computational models of associative learning. PMID- 11734856 TI - Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression. AB - Alteration of gene expression is a crucial component of adaptive responses to hypoxia. These responses are mediated by hypoxia-inducible transcription factors (HIFs). Here we describe an inhibitory PAS (Per/Arnt/Sim) domain protein, IPAS, which is a basic helix-loop-helix (bHLH)/PAS protein structurally related to HIFs. IPAS contains no endogenous transactivation function but demonstrates dominant negative regulation of HIF-mediated control of gene expression. Ectopic expression of IPAS in hepatoma cells selectively impairs induction of genes involved in adaptation to a hypoxic environment, notably the vascular endothelial growth factor (VEGF) gene, and results in retarded tumour growth and tumour vascular density in vivo. In mice, IPAS was predominantly expressed in Purkinje cells of the cerebellum and in corneal epithelium of the eye. Expression of IPAS in the cornea correlates with low levels of expression of the VEGF gene under hypoxic conditions. Application of an IPAS antisense oligonucleotide to the mouse cornea induced angiogenesis under normal oxygen conditions, and demonstrated hypoxia-dependent induction of VEGF gene expression in hypoxic corneal cells. These results indicate a previously unknown mechanism for negative regulation of angiogenesis and maintenance of an avascular phenotype. PMID- 11734857 TI - Extensive surface diversity of a commensal microorganism by multiple DNA inversions. AB - The dynamic interactions between a host and its intestinal microflora that lead to commensalism are unclear. Bacteria that colonize the intestinal tract do so despite the development of a specific immune response by the host. The mechanisms used by commensal organisms to circumvent this immune response have yet to be established. Here we demonstrate that the human colonic microorganism, Bacteroides fragilis, is able to modulate its surface antigenicity by producing at least eight distinct capsular polysaccharides-a number greater than any previously reported for a bacterium-and is able to regulate their expression in an on-off manner by the reversible inversion of DNA segments containing the promoters for their expression. This means of generating surface diversity allows the organism to exhibit a wide array of distinct surface polysaccharide combinations, and may have broad implications for how the predominant human colonic microorganisms, the Bacteroides species, maintain an ecological niche in the intestinal tract. PMID- 11734858 TI - Barttin is a Cl- channel beta-subunit crucial for renal Cl- reabsorption and inner ear K+ secretion. AB - Renal salt loss in Bartter's syndrome is caused by impaired transepithelial transport in the loop of Henle. Sodium chloride is taken up apically by the combined activity of NKCC2 (Na+-K--2Cl- cotransporters) and ROMK potassium channels. Chloride ions exit from the cell through basolateral ClC-Kb chloride channels. Mutations in the three corresponding genes have been identified that correspond to Bartter's syndrome types 1-3. The gene encoding the integral membrane protein barttin is mutated in a form of Bartter's syndrome that is associated with congenital deafness and renal failure. Here we show that barttin acts as an essential beta-subunit for ClC-Ka and ClC-Kb chloride channels, with which it colocalizes in basolateral membranes of renal tubules and of potassium secreting epithelia of the inner ear. Disease-causing mutations in either ClC-Kb or barttin compromise currents through heteromeric channels. Currents can be stimulated further by mutating a proline-tyrosine (PY) motif on barttin. This work describes the first known beta-subunit for CLC chloride channels and reveals that heteromers formed by ClC-K and barttin are crucial for renal salt reabsorption and potassium recycling in the inner ear. PMID- 11734859 TI - Isochorismate synthase is required to synthesize salicylic acid for plant defence. AB - Salicylic acid (SA) mediates plant defences against pathogens, accumulating in both infected and distal leaves in response to pathogen attack. Pathogenesis related gene expression and the synthesis of defensive compounds associated with both local and systemic acquired resistance (LAR and SAR) in plants require SA. In Arabidopsis, exogenous application of SA suffices to establish SAR, resulting in enhanced resistance to a variety of pathogens. However, despite its importance in plant defence against pathogens, SA biosynthesis is not well defined. Previous work has suggested that plants synthesize SA from phenylalanine; however, SA could still be produced when this pathway was inhibited, and the specific activity of radiolabelled SA in feeding experiments was often lower than expected. Some bacteria such as Pseudomonas aeruginosa synthesize SA using isochorismate synthase (ICS) and pyruvate lyase. Here we show, by cloning and characterizing an Arabidopsis defence-related gene (SID2) defined by mutation, that SA is synthesized from chorismate by means of ICS, and that SA made by this pathway is required for LAR and SAR responses. PMID- 11734861 TI - Pathophysiology, clinical manifestations and supportive care of metastatic brain cancer. AB - Brain metastases (BrM) are tumours that originate in tissues outside the central nervous system and spread secondarily to involve mainly the brain. The management of patients with cerebral metastases is complex, costly, and in some instances controversial. Furthermore, even in patients with widespread systemic cancer, the symptoms of the disease are often controllable while the symptoms of the BrM may be disabling. The treatment of BrM is one of the few areas of neuro-oncology where real progress has been made in the last twenty years. Moreover, the costs of managing this disease are rising, as therapies become more intensive and the number of patients with BrM increases. Modern neuroradiological imaging techniques, which are able to discover BrM earlier in the course of systemic cancer, and the greater efficacy of specific treatments, which lengthens survival, have increased the prevalence. The aggressive treatment of BrM may add some benefits to the patient, but its excessive cost leads to the necessity for accurate cost-effectiveness analysis. The latter begins with a complete understanding of the disease: its diagnosis, natural history and results of various modalities of treatment. While the development of BrM usually indicates a poor prognosis for the patient, advances in supportive care have made it possible to reverse most of the neurological symptoms and to give patients a meaningful extension of useful life. PMID- 11734862 TI - Prognostic factors in patients with brain metastases. AB - Management of patients with brain metastases (BrM) has been evolving over time, with a general tendency towards a more aggressive treatment approach. Although whole brain radiotherapy (WBRT) remains the mainstay of treatment for the majority of patients, the indications for stereotactic radiosurgery (SRS) with or without WBRT have been expanding rapidly. In order to select patients who would benefit from the improved intracranial control rates obtained with advanced radiation therapy techniques, knowledge of prognostic factors is essential. Literature findings indicate that performance status, age and systemic tumour activity (extracranial metastases and primary tumour status) are the major determinants of survival for patients with BrM. Based on recursive partitioning analysis (RPA) of these factors, the Radiation Therapy Oncology Group has developed a prognostic classification system for patients with BrM. RPA Class 1 represents the most favourable subset of patients with Karnofsky performance status of > or = 70, age under 65 years, controlled primary tumour and no extracranial metastases. RPA Class 3 patients have KPS < 70, associated with poor median survival rates. RPA Class 2 contains the remainder of patients, a rather heterogeneous subset of patients with intermediate survival rates. This RPA classification system, which has been validated for patients treated with WBRT, surgery and SRS, respectively, appears to be a valuable tool for assessment of prognosis in patients with BrM. PMID- 11734863 TI - Treatment of single brain metastases. AB - Brain metastases (BrM) occur in 30% to 50% of patients with malignant tumours and represent a relevant cause of morbidity and mortality. As documented by the extensive clinical use of magnetic resonance imaging with contrast enhancement, 25% to 35% of patients with BrM have a single localisation to the brain. Conventional therapeutic options for single BrM consist of either neurosurgery, or whole brain irradiation, or a combination of both. Treatment finality for the majority of the patients is palliation and life expectancy is in the order of a few months. In the last decade, the development of radiosurgery has represented an important advancement in the treatment of patients with BrM. This radiotherapeutic technique represents a possible non-invasive alternative to surgical exeresis for patients with lesions <3 cm, without a cystic component or obstructive hydrocephalus. Randomised clinical trials are warranted to define the appropriate indications and combinations of radiosurgery, neurosurgery, whole brain irradiation, and systemic treatment as well as the impact of different treatment modalities on amelioration of survival and quality of life. PMID- 11734864 TI - Controversies in the management of multiple brain metastases: the roles of radiosurgery and radiation therapy. AB - Multiple brain metastases (BrM) are a common challenge to patients with cancer. Tumour resection is used mainly for patients with large tumours that cause acute neurological syndromes. The prognosis, even after treatment with whole brain radiation therapy (WBRT), is poor with average expected survivals less than six months. For this reason, numerous centres have evaluated the role of stereotactic radiosurgery (SRS) in patients with solitary or multiple tumours. We conducted a randomised trial that compared radiosurgery plus WBRT to WBRT alone. The rate of local failure at one year was 100% after WBRT alone but only 8% in patients who had boost radiosurgery. The median time to local failure was six months after WBRT alone in comparison to 36 months after WBRT plus radiosurgery (p=0.0005). The median time to any brain failure was improved in the radiosurgery group (p=0.002). Survival was related to extent of extracranial disease (p=0.02). Combined WBRT and radiosurgery for patients with two to four BrM significantly improves control of brain disease. WBRT alone, for years the standard treatment, does not appear to provide lasting and effective care for most patients. Controversies remain in patient selection, number of BrM suitable for treatment, concomitant management of extracranial disease, and timing of therapy. PMID- 11734865 TI - Controversies in the management of brain metastases: the role of chemotherapy. AB - The role of chemotherapy (CT) for brain metastases (BrM) is still controversial. Limited life expectancy and presence of the blood-brain barrier (BBB) have been considered as contraindications for relatively aggressive therapies, such as CT. Nevertheless, more recent studies emphasise the possibility that in addition to historical nitrosoureas, also platinum derivatives, etoposide, teniposide, gemcitabine, irinotecan, topotecan and temozolomide can pass the BBB as they are active against selected BrM. Interaction of cytotoxic agents with other drugs may represent a problem in the treatment of BrM. By far, the most important factor conditioning the response to cytotoxic agents is the chemosensitivity of different tumours. CT proved to be effective in patients with BrM from lung cancer, mainly small cell lung cancer, breast cancer, choriocarcinoma and germ cell tumours. In these malignancies the responses to CT are similar to those observed in other metastatic sites. This observation, confirmed by many studies, contrasts with the theory, emphasised in the past, of the brain as a sanctuary. In fact BBB may be important in protecting normal brain tissue and micrometastases from CT, but not when BrM are symptomatic and clinically evident. The clue to the treatment of chemosensitive tumours is to assess the most active drugs and combinations. Cisplatin and etoposide was confirmed to be the treatment of choice in many situations (BrM from lung and also from breast cancer). Newer drugs are now on study: topotecan and temozolomide seem to be particularly promising (alone or in association with other drugs) in the treatment of various BrM (also from melanoma) and could represent an alternative to more widely-used drugs or as second-line treatments. PMID- 11734866 TI - Brain metastases in paediatric solid tumours. AB - Brain metastases in children with cancer are rare and their incidence is significantly lower (5-10%) than that reported in adults. The development of metastatic brain tumours in children is usually a manifestation of advanced disease and commonly occur after, or at the time of progression at other sites. This review summarises the salient clinical features of the most common paediatric solid tumours that metastasize to the brain including neuroblastoma, musculoskeletal sarcomas, germ cell tumours and melanoma. PMID- 11734867 TI - Treatment of ACL and PCL. PMID- 11734868 TI - Anterior and posterior cruciate ligament reconstruction in the new millennium: a global perspective. PMID- 11734869 TI - Biomechanical properties of quadruple tendon and patellar tendon femoral fixation techniques. AB - Two femoral fixation techniques for quadruple hamstring tendon grafts were compared under cyclic loading with the patellar tendon: the rectangular inserted pin (TransFix) and biodegradable interference screw fixation of the quadruple tendon and titanium interference screw fixation of the middle third of the patellar tendon. Porcine specimens were mounted onto a tension load machine, and the tendon-fixation-femur-complex was tested for stiffness, displacement during 800 cycles of loading between 50 and 250 N and ultimate tension load. TransFix fixation showed the greatest stiffness at 183.6 N/mm ( P<0.05). The least displacement under cyclic loading was observed for the titanium interference screw followed by the TransFix and biodegradable interference screw ( P<0.01). The ultimate tension load was greatest for the TransFix fixation at 1303+/-282 N, followed by patellar tendon fixation with 763+/-103 N and the biodegradable interference screw fixation with 480+/-133 N ( P<0.001). To reduce initial elongation of the graft and displacement at the fixation site, preconditioning of both the tendon and tendon-fixation complex is especially important when using quadruple tendons. TransFix fixation provides better stability and greater stiffness and pull-out strength than the other techniques. This finding is of clinical relevance to surgeons of the anterior cruciate ligament. PMID- 11734870 TI - C-reactive protein and erythrocyte sedimentation rate changes following arthroscopically assisted anterior cruciate ligament reconstruction. AB - We studied changes in C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) to establish normal values after uncomplicated anterior cruciate ligament (ACL) reconstruction. The levels of CRP and ESR were determined by serial measurements in 45 consecutively treated patients. Blood samples were collected before surgery and on postoperative days 1, 3, 7, 15, and 30. Both ESR and CRP showed a marked increase postoperatively, peaking between the 3rd and 7th postoperative days, the latter showing a faster return to normal ( P=0.286). These data show that CRP can be used as a more accurate predictor than ESR of postoperative complications if the blood level remains elevated or unexpectedly rises. PMID- 11734871 TI - Arthroscopic fixation of the fractures of the intercondylar eminence via transquadricipital tendinous portal. AB - We describe a new technique for arthroscopic reduction and internal fixation of fractures of the intercondylar eminence of the tibia. In this technique cannulated screws are placed through a new portal (transquadricipital tendinous) instead of anteromedial arthroscopic portal. Twelve patients who were treated with this technique for displaced types II and III fractures of the intercondylar eminence of the tibia were reviewed after a mean of 49 months. At follow-up all patients had excellent or good results, without any case of nonunion of the fracture or related complications, such as functional instability. Placement of cannulated screws through transquadricipital tendinous portal achieves fragment reduction easily, provides rigid fixation while avoiding arthrotomy, allowing early mobilization and return to activity. PMID- 11734872 TI - Localized pigmented villonodular synovitis of the knee: diagnostic challenge and arthroscopic treatment: a report of three cases. AB - The localized form of pigmented villonodular synovitis (LPVS) is a lesion characterized by focal involvement of the synovial membrane. The knee is the most commonly affected joint. We report three cases of LPVS of the knee which were not diagnosed upon clinical evaluation. The aim is to bring the attention of clinicians to this pathological entity, which is often regarded as extremely rare and is therefore not considered in the early differential diagnosis of various knee derangements. Diagnostic and therapeutic arthroscopy was performed. The lesions were completely resected and patohistological findings confirmed the diagnosis of LPVS. All of our three patients have remained asymptomatic at 8, 10, and 12-month follow-ups. PMID- 11734873 TI - Synovial hemangioma in Hoffa's fat pad (case report). AB - A 15-year-old girl presented with anterolateral knee pain for 6 months. There were tenderness and pain at the anterolateral triangulation of the knee joint. Magnetic resonance imaging revealed a tumor in the anterolateral corner of the knee joint. Arthroscopy did not show the tumor because it was located in fat pad. The tumor was found during fat pad shaving and was resected arthroscopically. Synovial hemangioma was demonstrated histologically. The patient is asymptomatic postoperatively. PMID- 11734874 TI - Health economics benefits following autologous chondrocyte transplantation for patients with focal chondral lesions of the knee. AB - Autologous chondrocyte transplantation (ACT) for the treatment of cartilage injuries has been in clinical use for several years. Since this new technique is potentially more costly and invasive than traditional conservative therapies, we evaluated the effect of ACT on clinical outcome, absenteeism, disability status, and total direct economic burden in 57 patients with full-thickness chondral lesions of the knee treated between 1987 and 1996. Patients graded good or excellent following ACT in the treatments groups were: femoral condyles (28/33), femoral condyles with anterior cruciate ligament (ACL) repair (5/5), osteochondritis dissecans (7/8), and patellar lesions (9/11). Pre-ACT, 57/57 patients were disabled and post-ACT (mean follow-up 7.3 years) 44/57 had no sickness, 10/57 had minor disability, and 1/57 was disabled. Two of the 57 patients suffered re-injury during the follow-up time. In the 10-year period prior to ACT, the average cost of absenteeism and surgery was SEK 982,457 ($ 122,807) and SEK 47,000 ($ 5,875), respectively, compared to the post-ACT period where both absenteeism and medical costs were dramatically reduced: SEK 9,508 ($ 1,189) and SEK 7,050 ($ 881), respectively. In conclusion, 49 of the 57 patients improved clinically as a result of the ACT treatment. A dramatic cost-saving effect was demonstrated over a projected 10-year period due to reduced absenteeism and disability. PMID- 11734875 TI - The presence of proprioceptive mechanoreceptors in the remnants of the ruptured ACL as a possible source of re-innervation of the ACL autograft. AB - This study investigated the presence of neural mechanoreceptors in the remnants of the ruptured ACL as a possible source of reinnervation of the ACL autologous graft. The remainder of the torn ACL was selected for further histological investigation from 17 patients during ACL reconstruction 3 months to 3.5 years after injury. Perioperatively two types of ACL remnant were identified. Fifteen patients had portions of ACL adapted at the PCL. In all of these patients we found mechanoreceptors (I and II). In five patients we found mushroomlike remnants which included either none or small numbers of mechanoreceptors. Free neural ends were found in both patient groups. There was a significant difference between the groups in regard to the mean number of mechanoreceptors I and II per slice. In conclusion, in patients with an ACL remnant adapted to the PCL, mechanoreceptors exist even 3 years after injury. If we accept that restoration of proprioception is the result of reinnervation of the ACL, leaving the ACL remnants as a source, if this is surgically possible without risk of Cyclop's lesion, may be of potential benefit to the patient. PMID- 11734876 TI - A pilot study on the relationship between physical impairment and activity restriction in persons with anterior cruciate ligament reconstruction at long term follow-up. AB - This study examined the relationship between impairment of the knee and activity restriction during activities of daily living and sports following anterior cruciate ligament reconstruction. Knee range of motion, swelling, pain, instability, ligamentous laxity, isometric and isokinetic muscle function and performance-based measures of activity restriction were measured in 44 subjects. Four measures of patient-reported activity restrictions, including the Activities of Daily Living, Sports Activities Scales of the Knee Outcome Survey, and global ratings of function during activities of daily life and sports, were statistically combined to create a composite variable representing the level of patient-reported activity restrictions for each subject. Hierarchical regression analysis revealed that 17% of the variability in patient-reported activity restrictions was accounted for by age, length of postoperative follow-up, and mechanism of injury. Addition of the one-legged hop, Lachman, anterior drawer, and varus stress tests accounted for an additional 40% of the variability of function. When pain and giving way were added to the model, 79% of the variability was explained. PMID- 11734877 TI - Arthroscopic vs. open Bankart reconstruction: a comparison using expected value decision analysis. AB - The reporting of midterm failure rates following arthroscopic shoulder stabilization for recurrent anterior instability of the glenohumeral joint has shown that the risk of failure may be higher with arthroscopic techniques than with traditional open methods. The use of expected value decision analysis offers an explicit, consistent, and structured means to assist the clinician in determining which stabilization technique to utilize. Decision analysis reveals that the surgical technique that is favored for a given patient depends on the value that the patient assigns to various potential outcomes following surgery, the perioperative morbidity of each of the two procedures, and the surgeon's probability of success with either technique PMID- 11734878 TI - Telemark skiing injuries: an 11-year study. AB - This study evaluated telemark injuries in a Swedish ski area in terms of injury ratio, location, and causes over time. During the seasons of 1989-2000 all injured telemark skiers ( n=94) who attended the medical center in Tarnaby, Sweden, within 48 h after the accident were registered and asked to fill in an injury form. A control group of noninjured telemark skiers were interviewed in the season of 1999-2000. The most common cause of injury was fall (70%) and the injury ratio was 1.2. There was a higher proportion of beginners in the injured population, and they had a fall/run ratio of 0.7, compared with 0.3 for average and advanced skiers. Ankle/foot injuries were most common (28% of injuries) followed by knee (20%) and head/neck (17%). The ankle/foot injuries decreased from 35% to 22% in the seasons 1989-1995 to 1995-2000. Beginners had more ankle/foot injuries than skilled participants. The severity of ankle/foot injuries classified as the Abbreviated Injury Scale group 2 or higher decreased from 33% to 21% during the study period. Twenty-seven percent used plastic and 73% leather boots. We found no association between boot material and ankle/foot injuries. The proportion of high boots with two or more buckles was 51%. High boots appeared to be protective against ankle/foot injuries. The proportion of high boots increased from 24% to 67% during the study period. Thus ankle/foot injuries were the most common injury location, but have decreased over time. The severity of these injuries has also decreased. A possible explanation could be the increased use of high boots. PMID- 11734879 TI - Radiofrequency energy induced heating of bovine articular cartilage: comparison between temperature-controlled, monopolar, and bipolar systems. AB - This in vitro investigation characterized temperature changes associated with radiofrequency (RF) energy induced heating of bovine articular cartilage using temperature-controlled, monopolar (Vulcan RF system and Vulcan, TAC-S Electrothermal Probe) and bipolar (VAPR II RF system and VAPR TC RF electrode) electrosurgical equipment. The RF generators were used at the same setting (set temperature 70 degrees C; 30 W). The cartilage tissue sample was placed in a saline bath maintained at room temperature. Temperatures were recorded using fluoroptic thermometry at the RF electrode-tissue interface at 1-s intervals before, during deliver of RF energy (1- and 2-s), and after (1- to 3-s). For both electrosurgical systems the mean RF electrode-tissue interface temperatures were significantly ( P<0.05) higher than the mean baseline value during delivery of RF energy (monopolar, highest mean temperature, 65.7 degrees C; bipolar, highest mean temperature, 54.1 degrees C). In general, during and after the deliver of RF energy, the monopolar RF system produced tissue temperatures that were significantly ( P<0.05) higher than those produced by the bipolar RF system. Neither electrosurgical system exceeded the set temperature of 70 degrees C. These findings provide basic tissue temperature characteristics for the newly developed, temperature-controlled RF devices applied to articular cartilage. PMID- 11734881 TI - Thermoregulated expression of virulence factors in plant-associated bacteria. AB - Pathogenic bacteria with habitats inside and outside a given host react to changes in environmental parameters by synthesizing gene products specifically needed during pathogenic or saprophytic growth. Temperature effects have been investigated in detail for pathogens of warm-blooded hosts, and major principles governing the temperature-sensing mechanism have been uncovered. Generally, transcription of virulence genes in these pathogens is induced at higher temperatures (37-41 degrees C), which are typical for body cavities and host tissues. However, effects of temperature on virulence determinants in plant pathogenic bacteria have not been focused on in detail. Interestingly, almost all virulence genes of plant pathogenic bacteria studied with respect to temperature exhibit increased transcription at temperatures well below the respective growth optima. This includes virulence determinants such as those directing bacteria-to plant gene transfer, plant cell-wall-degrading enzymes, phytotoxins, ice nucleation activity, exopolysaccharide production, and the type III protein secretion machinery. Although many of the studied phytopathogens cause "cold weather" diseases, the ecological rationale for this phenomenon remains to be studied in detail. This mini-review summarizes our current knowledge on thermoregulation of cellular processes taking place in bacterial phytopathogens in response to temperature changes. Since the temperature range of interest is different from that relevant to pathogens of mammals, one envisions novel principles of thermo-sensing in bacteria interacting with plants. PMID- 11734882 TI - Biliproteins and phycobilisomes from cyanobacteria and red algae at the extremes of habitat. AB - This review considers the properties of biliproteins from cyanobacteria and red algae that grow in extreme habitats. Three situations are presented: cyanobacteria that grow at high temperatures; a red alga that grows in acidic conditions at high temperature; and an Antarctic red alga that grows in the cold in dim light conditions. In particular, the properties of their biliproteins are compared to those from organisms from more usual environments. C-phycocyanins from two cyanobacteria able to grow at high temperatures are found to differ in their stabilities when compared to C-phycocyanin from mesophilic algae. They differ in opposite ways, however. One is more stable to dissociation than the mesophilic protein, and the other is more easily dissociated at low temperatures. The thermophilic proteins resist thermal denaturation much better than the mesophilic proteins. The most thermophilic cyanobacterium has a C-phycocyanin with a unique blue-shifted absorption maximum which does not appear to be part of the adaptation of the cyanobacterium to high temperature. The C-phycocyanin from the high-temperature red alga is able to resist dissociation better than mesophilic C-phycocyanins. Electron micrographs show the phycobilisomes of these algae. The Antarctic alga grows under ice at some distance down the water column. Its R-phycoerythrin has a novel absorption spectrum that gives the alga an improved ability to harvest blue light. This may enhance its survival in its light-deprived habitat. PMID- 11734883 TI - Alkanesulfonate degradation by novel strains of Achromobacter xylosoxidans, Tsukamurella wratislaviensis and Rhodococcus sp., and evidence for an ethanesulfonate monooxygenase in A. xylosoxidans strain AE4. AB - Novel isolates of Achromobacter xylosoxidans, Tsukamurella wratislaviensis and a Rhodococcus sp. are described. These grew with short-chain alkanesulfonates as their sole source of carbon and energy. T. wratislaviensis strain SB2 grew well with C(3)-C(6) linear alkanesulfonates, isethionate and taurine, Rhodococcus sp. strain CB1 used C(3)-C(10) linear alkanesulfonates, taurine and cysteate, but neither strain grew with ethanesulfonate. In contrast, A. xylosoxidans strain AE4 grew well with ethanesulfonate, making it the first bacterium to be described which can grow with this compound. It also grew with unsubstituted C(3)-C(5) alkanesulfonates and isethionate. Hydrolysis was excluded as a mechanism for alkanesulfonate metabolism in these strains; and evidence is given for a diversity of uptake and desulfonatase systems. We provide evidence for an initial monooxygenase-dependent desulfonation in the metabolism of ethanesulfonate and propanesulfonate by A. xylosoxidans strain AE4. PMID- 11734884 TI - Regulation of the synthesis of cyclic glucan in Xanthomonas campestris by a diffusible signal molecule. AB - The rpf gene cluster of Xanthomonas campestris pv. campestris is involved in the co-ordinate positive regulation of the production of extracellular enzymes and the extracellular polysaccharide xanthan. Several of the rpf genes are involved in a regulatory system involving the small diffusible molecule DSF (for diffusible signal factor). Synthesis of DSF requires RpfF, and a two-component sensory transduction system involving RpfC has been implicated in the perception of the signal and signal transduction. Here we show that mutations in both rpfF and rpfC lead to reductions in the levels of cyclic glucan. The levels of cyclic glucan synthetase in membrane preparations from rpfF and rpfC mutants were, however, unaltered from the wild-type. Similar alterations in the level of cyclic glucan without changes in cyclic glucan synthetase activity were seen when wild type bacteria were exposed to osmotic stress. These results extend the range of cellular functions subject to regulation by the rpf genes and DSF system. PMID- 11734885 TI - Purification and properties of two chitinolytic enzymes of Serratia plymuthica HRO-C48. AB - The chitinolytic rhizobacterium Serratia plymuthica HRO-C48 was previously selected as a biocontrol agent of phytopathogenic fungi. One endochitinase (E.C. 3.2.1.14), CHIT60, and one N-acetyl-beta-1,4- D-hexosaminidase (E.C. 3.2.1.52), CHIT100, were purified and characterized. The endochitinase CHIT60, with an apparent molecular mass of 60.5 kDa, had a N-terminal amino acid sequence highly similar to that of chitinases A from Serratia liquefaciens and Serratia marcescens. The enzyme activity had its peak at 55 degrees C and pH 5.4, and increased by more than 20% in the presence of 10 mM Ca(2+), Co(2+) or Mn(2+). Activity was inhibited by 80% in the presence of 10 mM Cu(2+). CHIT100 appeared to be a monomeric enzyme with a molecular mass of 95.6 kDa and a pI of 6.8. Optimal activity was obtained at 43 degrees C and pH 6.6, and decreased by more than 90 % in the presence of 10 mM Co(2+) or Cu(2+). CHIT100 (100 microg ml(-1)) inhibited spore germination and germ tube elongation of the phytopathogenic fungus Botrytis cinerea by 28 % and 31.6 %, respectively. With CHIT60 (100 microg ml(-1)), the effect was more pronounced: 78 % inhibition of of germination and 63.9 % inhibition of germ tube elongation. PMID- 11734886 TI - Acid phosphatases of Metarhizium anisopliae during infection of the tobacco hornworm Manduca sexta. AB - Three acid phosphatase (AcP) isozymes, pI 8.1, 8.0 and 7.8, were isolated, purified and partially characterised from optimised cultures of the entomopathogenic fungus Metarhizium anisopliae. The enzymes had similar molecular masses (approximately 44.0 kDa), and could degrade sugar phosphates found in the haemolymph of a host insect, the tobacco hornworm Manduca sexta. The AcP activity in haemolymph of mycosed insects increased significantly over controls, and some new isozymes were present. The infection-related isoforms were similar in molecular mass and pI to some of the in vitro AcP isozymes of M. anisopliae. Results of dot blot and Western blot analyses using anti-AcP antibodies suggested that at least one Metarhizium phosphatase isoform was present in haemolymph of infected caterpillars. Antibodies did not cross-react with immune (chemically stimulated) or non-immune haemolymph from Manduca sexta. Consistent with the appearance of highly active fungal phosphatase in caterpillar blood, free phosphate concentration increased dramatically during the late stages of infection to a level two to five times that of controls. Phosphate was limiting to growth of the fungus at the concentration found in control haemolymph and supplementation of phosphate significantly increased fungal growth in vitro in haemolymph. These results suggest that Metarhizium AcP may play a key role in providing phosphorus for fungal growth at the expense of the insect. PMID- 11734887 TI - Monoalkylether phospholipids in the sulfate-reducing bacteria Desulfosarcina variabilis and Desulforhabdus amnigenus. AB - In this study, cellular lipid compositions of two mesophilic sulfate-reducing bacteria were analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS). In Desulfosarcina variabilis and Desulforhabdus amnigenus, alkylether-containing phospholipids were detected which had previously only been found in significant amounts in deeply branching hyperthermophilic bacteria and archaea. Combining information from HPLC-MS analysis and chemical degradation experiments, ether lipids were identified as 1-alkyl-2-acyl phosphatidyl ethanolamines, glycerols and cholines. In Desulforhabdus amnigenus, n-penta-, n-hexa- and n-heptadecyl ethers were present (in order of decreasing abundance), whereas Desulfosarcina variabilis solely contained n-hexadecyl ether side chains. PMID- 11734888 TI - Pyruvate decarboxylase: a key enzyme for the oxidative metabolism of lactic acid by Acetobacter pasteurianus. AB - Acetobacter pasteurianus, an obligately oxidative bacterium, is the first organism shown to utilize pyruvate decarboxylase (PDC) as a central enzyme for oxidative metabolism. In plants, yeast, and other bacteria, PDC functions solely as part of the fermentative ethanol pathway. During the growth of A. pasteurianus on lactic acid, the central intermediate pyruvate is cleaved to acetaldehyde and CO(2) by PDC. Acetaldehyde is subsequently oxidized to its final product, acetic acid. The presence of the PDC enzyme in A. pasteurianus was confirmed by zymograms stained for acetaldehyde production, enzyme assays using alcohol dehydrogenase as the coupling enzyme, and by cloning and characterization of the pdc operon. A. pasteurianus pdc was also expressed in recombinant Escherichia coli. The level of PDC activity was regulated in response to growth substrate, highest with lactic acid and absent with mannitol. The translated PDC sequence (548 amino acids) was most similar to that of Zymomonas mobilis, an obligately fermentative bacterium. A second operon ( aldA) was also found which is transcribed divergently from pdc. This operon encodes a putative aldehyde dehydrogenase (ALD2; 357 amino acids) related to class III alcohol dehydrogenases and most similar to glutathione-dependent formaldehyde dehydrogenases from alpha Proteobacteria and Anabeana azollae. PMID- 11734889 TI - Nonribosomal peptide synthetase genes occur in most cyanobacterial genera as evidenced by their distribution in axenic strains of the PCC. AB - Previous studies largely carried out with environmental samples or axenic and non axenic cultures suggested that cyanobacteria may be a rich source of hitherto unexplored bioactive compounds. This has been confirmed in the present study by a screening of 146 axenic strains from the Pasteur Culture Collection (PCC) of cyanobacteria. Use of degenerate PCR primers, designed on the basis of conserved sequence motifs in the aminoacyl-adenylation domain of peptide synthetases, revealed the presence of the corresponding genes in the majority (75.3%) of the strains examined. Among unicellular cyanobacteria, only Chamaesiphon sp. strain PCC 6605, two strains of Gloeocapsa and most Microcystis isolates (22 out of 24) contained these genes; no amplicons were detected for any members of the genera Cyanothece, Gloeobacter and Gloeothece and the genetically diverse representatives of Synechococcus and Synechocystis. By contrast, eight out of ten pleurocapsalean members, 16 out of 25 oscillatorian strains, and all but two of the 63 filamentous heterocystous cyanobacteria tested gave positive amplification results. This information will be highly valuable for further exploring the corresponding cyanobacterial peptides and for elucidating the bioactivity of such non-ribosomally synthesized molecules. PMID- 11734891 TI - Dependence of linkage of alleles on their physical distance in natural transformation of Acinetobacter sp. strain ADP1. AB - The interdependence of genetic linkage in transformation and physical distance was studied in the bacterium Acinetobacter sp. strain ADP1. Transformation experiments were performed using 17 strains containing different mutations within the 21-kb pca-qui-pob gene cluster as recipients for the DNA of one of two strains carrying a mutation causing a temperature-sensitive phenotype. The different phenotypes of the transformants (temperature-sensitive or wild-type like) were used to evaluate linkage. Combination of the recipient and donor strains resulted in physical distances ranging from 2 bp to 10,533 bp. A logarithmic relationship of decreasing linkage and increasing distance was observed, thus leading to calibration of a system for analysis of physical distance derived from linkage data. Limitations of this application are described here: Certain mutations (3 out of 17 mutations used in this study) are an exception to the observed relationship and result in much lower linkage than expected. Observed DNA sequence repetitions leading to DNA rearrangements may be the cause of this anomaly. PMID- 11734890 TI - Fatty-acid-displaced transcriptional repressor, a conserved regulator of cytochrome P450 102 transcription in Bacillus species. AB - Bacillus subtilis strain 168 encodes two flavocytochromes P450, Cyp102A2 and Cyp102A3. The cyp102A3 gene is preceded by, and organized in an operon with, a gene for a transcriptional regulator, encoded by fatR. The paralogous gene, cyp102A2, is most likely transcribed as a mono-cistronic message. We show that fatR encodes a protein that binds to an operator sequence that is present upstream of its own reading frame, thereby repressing the expression of the fatR cyp102A3 operon. Unsaturated fatty acids and phytanic acid have the capacity to interact with FatR and to abrogate its binding to the operator sequence. PMID- 11734892 TI - Selective pressures on RNA hairpins in vivo and in vitro. AB - Comparison of the most stable potential hairpins in the sequences of natural ribozymes with those in the randomized sequences has revealed that the hairpin loop energies are lower than expected by chance. Although these hairpins are not necessarily parts of functional structures, there is a selective pressure to diminish the destabilizing free energies of the hairpin loops. In contrast, no significant bias is observed in the stacking values of the most stable stems. In the ribozymes isolated in vitro the loops of potential hairpins are closer to random values, which can result in less efficient folding rates. Furthermore, the effects of kinetic traps seem to be more significant in the folding pathways of the in vitro isolates due to a potential to form stable stacks incompatible with the functional folds. Similarly to natural ribozyme sequences, the untranslated regions of viral RNAs also form hairpins with relatively low loop free energies. These evolutionary trends suggest ways for efficient engineering of improved RNA constructs on the basis of analysis of in vitro isolates and approaches for the search of regions coding for functional RNA structures in large genome sequences. PMID- 11734893 TI - SINE retrotransposition during the evolution of the Pecoran ruminants. AB - SINE retrotransposition events have proven their value as phylogenetic markers in several eukaryotic taxa at different taxonomic levels. The genomes of ruminants contain three related SINE elements, Bov-tA, Bov-A2, and Bov-B. To estimate the time points of retrotransposition of individual copies of these SINEs, we designed PCR primers on database sequences containing SINE insertions in cattle, sheep, or goat genomes and tested for the presence of these copies in the genomes of other ruminants. It was checked by sequencing whether length variation of the PCR products reflected a SINE retrotransposition. One Bov-B and nine Bov-tA insertions were shared by cattle, sheep, goat, and giraffe, indicating an early retrotransposition event before the radiation of the Pecora, while three other Bov-tA and two Bov-B elements were apparently inserted later. The ruminant alpha lactalbumine gene contains a hotspot of early and more recent Bov-tA insertions, a Bov-tA replacement as well as a recent Bov-B insertion. Three Bov-A2 insertions were found to be shared only by the Bovidae, the Bovini, and the Bos and Bison species, respectively, indicating that most Bov-A2 insertions are relatively recent. The time elapsed since the retrotransposition was also reflected in the degeneration of the direct repeats that flank SINE inserts. We suggest that retrotransposition of SINEs may serve as phylogenetic markers in the ruminant families, subfamilies, and even tribes. In addition, sequencing of SINE insertions revealed several other unique deletions/insertions that also may be informative for phylogenetic reconstructions of ruminants. PMID- 11734894 TI - Classification and phylogenetic analysis of the cAMP-dependent protein kinase regulatory subunit family. AB - The members of the PKA regulatory subunit family (PKA-R family) were analyzed by multiple sequence alignment and clustering based on phylogenetic tree construction. According to the phylogenetic trees generated from multiple sequence alignment of the complete sequences, the PKA-R family was divided into four subfamilies (types I to IV). Members of each subfamily were exclusively from animals (types I and II), fungi (type III), and alveolates (type IV). Application of the same methodology to the cAMP-binding domains, and subsequently to the region delimited by beta-strands 6 and 7 of the crystal structures of bovine RIalpha and rat RIIbeta (the phosphate-binding cassette; PBC), proved that this highly conserved region was enough to classify unequivocally the members of the PKA-R family. A single signature sequence, F-G-E-[LIV]-A-L-[LIMV]-x(3)-[PV]-R [ANQV]-A, corresponding to the PBC was identified which is characteristic of the PKA-R family and is sufficient to distinguish it from other members of the cyclic nucleotide-binding protein superfamily. Specific determinants for the A and B domains of each R-subunit type were also identified. Conserved residues defining the signature motif are important for interaction with cAMP or for positioning the residues that directly interact with cAMP. Conversely, residues that define subfamilies or domain types are not conserved and are mostly located on the loop that connects alpha-helix B' and beta strand 7. PMID- 11734895 TI - The evolutionary history of prosaposin: two successive tandem-duplication events gave rise to the four saposin domains in vertebrates. AB - Prosaposin is a multifunctional protein encoded by a single-copy gene. It contains four saposin domains (A, B, C, and D) occurring as tandem repeats connected by linker sequences. Because the saposin domains are similar to one another, it is deduced that they were created by sequential duplications of an ancestral domain. There are two types of evolutionary scenarios that may explain the creation of the four-domain gene: (1) two rounds of tandem internal gene duplication and (2) three rounds of duplications. An evolutionary and phylogenetic analysis of saposin DNA and amino acid sequences from human, mouse, rat, chicken, and zebrafish indicates that the first evolutionary scenario is the most likely. Accordingly, an ancestral saposin-unit duplication produced a two domain gene, which, subsequently, underwent a second complete tandem duplication to give rise to the present four-domain structure of the prosaposin gene. PMID- 11734896 TI - Correlations between mRNA expression levels and GC contents of coding and untranslated regions of genes in rodents. AB - Gene expression is regulated by a highly coordinated network of events whose efficiency may constrain the level of expression. Among other factors, natural selection for increased translational efficiency and/or fidelity may shape nucleotide composition and, hence, codon usage during evolution. Previous studies have shown that highly expressed genes in Saccharomyces cerevisiae, Caenorhabditis elegans, and Drosophila melanogaster have relatively higher codon usage biases. However, in the case of mammals, results have been equivocal. In this study, we assessed the correlation between nucleotide composition and mRNA expression levels of rodent genes measured by cDNA microarray and serial analysis of gene expression (SAGE) techniques. We found that mRNA expression levels were correlated with the third nucleotide position GC (GC3) content for both Rattus norvegicus (r = 0.246, p = 0.01; N = 110) and Mus musculus (r = 0.21, p = 0.0026; N = 203) genes. However, no significant correlation was evident between mRNA expression level and GC contents of 5'- and 3'-untranslated regions (UTRs) for either species. This suggests that, in rodents, nucleotide composition of coding sequences and UTRs might evolve differentially when considered along an expression gradient. Accordingly, it is possible that higher GC levels may present the rodent genes with a selective advantage for translational efficiency. However, the increase in GC3 content seems to level off above an expressional threshold (e.g., >or=threefold the median expression for R. norvegicus), suggesting that conflicting demands posed by different aspects of transcriptional and translational machineries (e.g., efficiency versus fidelity) may set an upper limit for GC3. PMID- 11734897 TI - Maximum likelihood methods reveal conservation of function among closely related kinesin families. AB - We have reconstructed the evolution of the anciently derived kinesin superfamily using various alignment and tree-building methods. In addition to classifying previously described kinesins from protists, fungi, and animals, we analyzed a variety of kinesin sequences from the plant kingdom including 12 from Zea mays and 29 from Arabidopsis thaliana. Also included in our data set were four sequences from the anciently diverged amitochondriate protist Giardia lamblia. The overall topology of the best tree we found is more likely than previously reported topologies and allows us to make the following new observations: (1) kinesins involved in chromosome movement including MCAK, chromokinesin, and CENP E may be descended from a single ancestor; (2) kinesins that form complex oligomers are limited to a monophyletic group of families; (3) kinesins that crosslink antiparallel microtubules at the spindle midzone including BIMC, MKLP, and CENP-E are closely related; (4) Drosophila NOD and human KID group with other characterized chromokinesins; and (5) Saccharomyces SMY1 groups with kinesin-I sequences, forming a family of kinesins capable of class V myosin interactions. In addition, we found that one monophyletic clade composed exclusively of sequences with a C-terminal motor domain contains all known minus end-directed kinesins. PMID- 11734898 TI - Comparison of substitution rates in ZFX and ZFY introns of sheep and goat related species supports the hypothesis of male-biased mutation rates. AB - There is a growing body of evidence that males serve as the major generators of mutations, due to the larger number of cell divisions involved in sperm compared to egg production. In mammals, this hypothesis (referred to as "male-driven evolution") has been tested by comparison of nucleotide substitution rates on the X and Y sex chromosomes in a limited number of taxa, predominantly primates and rodents. This study asks whether male-driven evolution is a more general phenomenon among mammals, by comparison of paralogous ZFX and ZFY intron sequences in sheep and goat species (the tribe Caprini). The male-to-female mutation ratio, alpha(m), was estimated to be between 2.93 (95% CI, 1.51-8.61) and 3.94 (95% CI, 1.25-32.29) when calculated using pairwise distance and branch length, respectively, suggesting that the Caprini are subject to weak, male driven evolution. Comparison to published values for primates, felids, and rodents implies that there may be some correlation with reproductive life span. However, this is difficult to test with current data because confidence intervals are large and overlapping. Nonindependent evolution of paralogous sequences and/or the presence of selective constraints could lead to inaccurate estimates of alpha(m). No evidence for gene conversion between the ZFX and the ZFY introns was found, and this suggests that they have evolved independently during the radiation of the Caprini. Finally, there was no apparent evidence that these introns are subject to selective constraints, although low levels of intraspecific polymorphism reduce the power of neutrality tests. PMID- 11734899 TI - Hepatitis C virus evolutionary patterns studied through analysis of full-genome sequences. AB - The evolutionary patterns of hepatitis C virus (HCV), including the best-fitting nucleotide substitution model and the molecular clock hypothesis, were investigated by analyzing full-genome sequences available in the HCV database. The likelihood ratio test allowed us to discriminate among different evolutionary hypotheses. The phylogeny of the six major HCV types was accurately inferred, and the final tree was rooted by reconstructing the hypothetical HCV common ancestor with the maximum likelihood method. The presence of phylogenetic noise and the relative nucleotide substitution rates in the different HCV genes were also examined. These results offer a general guideline for the future of HCV phylogenetic analysis and also provide important insights on HCV origin and evolution. PMID- 11734900 TI - Phylogenetic analysis of 18S rRNA and the mitochondrial genomes of the wombat, Vombatus ursinus, and the spiny anteater, Tachyglossus aculeatus: increased support for the Marsupionta hypothesis. AB - The monotremes, the duck-billed platypus and the echidnas, are characterized by a number of unique morphological characteristics, which have led to the common belief that they represent the living survivors of an ancestral stock of mammals. Analysis of new data from the complete mitochondrial (mt) genomes of a second monotreme, the spiny anteater, and another marsupial, the wombat, yielded clear support for the Marsupionta hypothesis. According to this hypothesis marsupials are more closely related to monotremes than to eutherians, consistent with a basal split between eutherians and marsupials/monotremes among extant mammals. This finding was also supported by analysis of new sequences from a nuclear gene- 18S rRNA. The mt genome of the wombat shares some unique features with previously described marsupial mtDNAs (tRNA rearrangement, a missing tRNA(Lys), and evidence for RNA editing of the tRNA(Asp)). Molecular estimates of genetic divergence suggest that the divergence between the platypus and the spiny anteater took place approximately 34 million years before present (MYBP), and that between South American and Australian marsupials approximately 72 MYBP. PMID- 11734901 TI - Ascidian and amphioxus Adh genes correlate functional and molecular features of the ADH family expansion during vertebrate evolution. AB - The alcohol dehydrogenase (ADH) family has evolved into at least eight ADH classes during vertebrate evolution. We have characterized three prevertebrate forms of the parent enzyme of this family, including one from an urochordate (Ciona intestinalis) and two from cephalochordates (Branchiostoma floridae and Branchiostoma lanceolatum). An evolutionary analysis of the family was performed gathering data from protein and gene structures, exon-intron distribution, and functional features through chordate lines. Our data strongly support that the ADH family expansion occurred 500 million years ago, after the cephalochordate/vertebrate split, probably in the gnathostome subphylum line of the vertebrates. Evolutionary rates differ between the ancestral, ADH3 (glutathione-dependent formaldehyde dehydrogenase), and the emerging forms, including the classical alcohol dehydrogenase, ADH1, which has an evolutionary rate 3.6-fold that of the ADH3 form. Phylogenetic analysis and chromosomal mapping of the vertebrate Adh gene cluster suggest that family expansion took place by tandem duplications, probably concurrent with the extensive isoform burst observed before the fish/tetrapode split, rather than through the large scale genome duplications also postulated in early vertebrate evolution. The absence of multifunctionality in lower chordate ADHs and the structures compared argue in favor of the acquisition of new functions in vertebrate ADH classes. Finally, comparison between B. floridae and B. lanceolatum Adhs provides the first estimate for a cephalochordate speciation, 190 million years ago, probably concomitant with the beginning of the drifting of major land masses from the Pangea. PMID- 11734902 TI - Origin and evolution of viral interleukin-10 and other DNA virus genes with vertebrate homologues. AB - Phylogenies of gene families including members in both vertebrates and DNA viruses of the poxvirus and/or herpesvirus families showed that the viral genes originated at widely different times over the history of life. Certain of these viral genes (for example, the genes encoding the large and small subunits of ribonucleoside-diphosphate reductase) originated before animals diverged from fungi, while others originated much more recently. The most striking examples of recent origin involved viral genes encoding the cytokine interleukin-10 (IL-10), which originated independently in viruses at least three times since the divergence of the orders of eutherian mammals, presumably by viral capture of host genes. In certain domains, viral IL-10 genes showed significantly higher rates of nonsynonymous substitution than their nearest mammalian homologues. Though the mutation rate in these viral genes is up to 20 times that of the corresponding mammalian genes, a high mutation rate alone did not account for these differences because they were not seen in all domains. Rather, in certain domains it appears that functional constraints present in the case of mammalian IL-10 are relaxed in the case of the viral homologues. Furthermore, a nonrandom pattern of change with respect to amino acid residue charge in the N-terminal portion of the mature protein has occurred repeatedly in independently derived viral IL-10 genes, strongly suggesting that positive selection has led to divergence of this functionally important domain in viral IL-10. PMID- 11734903 TI - Age and detection of retroprocessed pseudogenes in murine rodents. AB - Retroprocessed pseudogenes, calmodulin II (psi1, psi2, and psi3 CALMII), psi alpha-tubulin, pi-glutathione S-transferase (psi pi-GST) from rat, lactic acid dehydrogenase (psi LDH) from mouse, and heat shock protein 60 chaperonin (psi HSP60) from Chinese hamster, were examined for their presence in these species by polymerase chain reaction (PCR). Pseudogenes of these murine rodents were detected by PCR only in those species in which the genes were originally identified, suggesting that the selected pseudogene of one species arose too recently to be detected in the genomes of the other rodent species. The calculated ages of the rodent pseudogenes ranged from 1.7 Myr (psi alpha-tubulin) to 7.5 Myr (psi3 CALMII) when employing a homologous functional gene of the taxon as a reference in the relative rate test with the mouse or rat as the outgroup. Given the high rate of divergence of the genes of rodents relative to other species, selection of an outgroup with similar mutation rates seems warranted. To justify further the conclusion that the selected pseudogenes were indeed retroprocessed after these three taxa diverged, the presence of the pseudogenes in the genome of different rat species was examined. The existence of psi3 CALMII and psi alpha-tubulin pseudogenes of Rattus norvegicus among species belonging to Rattus sensu stricto is evidence for the common ancestry of this group. PMID- 11734904 TI - Phylogenetic relationships and biochemical properties of the duplicated cytosolic and mitochondrial isoforms of malate dehydrogenase from a teleost fish, Sphyraena idiastes. AB - Unlike birds and mammals, teleost fish express two paralogous isoforms (paralogues) of cytosolic malate dehydrogenase (cMDH; EC 1.1.1.37; NAD+: malate oxidoreductase) whose evolutionary relationships to the single cMDH of tetrapods are unknown. We sequenced complementary DNAs for both cMDHs and the mitochondrial isoform (mMDH) of the fish Sphyraena idiastes (south temperate barracuda) and compared the sequences, kinetic properties, and thermal stabilities of the three isoforms with those of mammalian orthologues. Both fish cMDHs comprise 333 residues and have subunit masses of approximately 36 kDa. One cytosolic isoform, cMDH-S, was significantly more heat-stable than either the other cMDH (cMDH-L) or mMDH. In contradiction to the generally accepted model of vertebrate cMDH evolution, our phylogenetic analysis indicates that the duplication of the fish cytosolic paralogues occurred after the divergence of the lineages leading to teleosts and tetrapods. cMDH-L and cMDH-S differed in optimal concentrations of substrates and cofactors and apparent Michaelis-Menten constants, suggesting that the two paralogues may play distinct physiological roles. Differences in intrinsic thermal stability among MDH paralogues may reflect different degrees of stabilization in vivo by extrinsic stabilizers, notably protein concentration in the case of mMDH. Thermal stabilities of porcine mMDH and cMDH-L, but not cMDH-S, were significantly increased when denaturation was measured at a high protein (bovine serum albumin; BSA) concentration, but the BSA-induced stabilization reduced the catalytic activity. PMID- 11734905 TI - Dynamic insertion-deletion of introns in deuterostome EF-1alpha genes. AB - To test the validity of intron-exon structure as a phylogenetic marker, the intron-exon structure of EF-1alpha genes was investigated for starfish, acornworms, ascidians, larvaceans, and amphioxus and compared with that of vertebrates. Of the 11 distinct intron insertion sites found within the coding regions of the deuterostome EF-1alpha genes, 7 are shared by several taxa, while the remainder are unique to certain taxa. Examination of the shared introns of the deuterostome EF-1alpha gene revealed that independent intron loss or intron insertion must have occurred in separate lineages of the deuterostome taxa. Maximum parsimony analysis of the intron-exon data matrix recovered five parsimonious trees (consistency index = 0.867). From this result, we concluded that the intron-exon structure of deuterostome EF-1alpha has evolved more dynamically than previously thought, rendering it unsuitable as a phylogenetic marker. We also reconstructed an evolutionary history of intron insertion deletion events on the deuterostome phylogeny, based on several molecular phylogenetic studies. These analyses revealed that the deuterostome EF-1alpha gene has lost individual introns more frequently than all introns simultaneously. PMID- 11734906 TI - Expected relationship between the silent substitution rate and the GC content: implications for the evolution of isochores. AB - The relationship between the silent substitution rate (Ks) and the GC content along the genome is a focal point of the debate about the origin of the isochore structure in vertebrates. Recent estimation of the silent substitution rate showed a positive correlation between Ks and GC content, in contradiction with the predictions of both the regional mutation bias model and the selection or biased gene conversion model. The aim of this paper is to help resolve this contradiction between theoretical studies and data. We analyzed the relationship between Ks and GC content under (1) uniform mutation bias, (2) a regional mutation bias, and (3) mutation bias and selection. We report that an increase in Ks with GC content is expected under mutation bias because of either nonequilibrium of the isochore structure or an increasing mutation rate from AT toward GC nucleotides in GC-richer isochores. We show by simulations that CpG deamination tends to increase the mutation rate with GC content in a regional mutation bias model. We also demonstrate that the relationship between Ks and GC under the selectionist or biased gene conversion model is positive under weak selection if the mutation selection equilibrium GC frequency is less than 0.5. PMID- 11734907 TI - Curiously modern DNA for a "250 million-year-old" bacterium. AB - Studies of ancient DNA have attracted considerable attention in scientific journals and the popular press. Several of the more extreme claims for ancient DNA have been questioned on biochemical grounds (i.e., DNA surviving longer than expected) and evolutionary grounds (i.e., nucleotide substitution patterns not matching theoretical expectations for ancient DNA). A recent letter to Nature from Vreeland et al. (2000), however, tops all others with respect to age and condition of the specimen. These researchers extracted and cultured a bacterium from an inclusion body from what they claim is a 250 million-year (Myr)-old salt crystal. If substantiated, this observation could fundamentally alter views about bacterial physiology, ecology and evolution. Here we report on molecular evolutionary analyses of the 16S rDNA from this specimen. We find that 2-9-3 differs from a modern halophile, Salibacillus marismortui, by just 3 unambiguous bp in 16S rDNA, versus the approximately 59 bp that would be expected if these bacteria evolved at the same rate as other bacteria. We show, using a Poisson distribution, that unless it can be shown that S. marismortui evolves 5 to 10 times more slowly than other bacteria for which 16S rDNA substitution rates have been established, Vreeland et al.'s claim would be rejected at the 0.05 level. Also, a molecular clock test and a relative rates test fail to substantiate Vreeland et al.'s claim that strain 2-9-3 is a 250-Myr-old bacterium. The report of Vreeland et al. thus falls into a long series of suspect ancient DNA studies. PMID- 11734908 TI - Testing claims of gene conversion between multigene family members: examples from echinoderm actin genes. AB - Molecular genetics studies often infer the occurrence of gene conversion events based on simple sequence similarity observations that do not include any statistical analyses. I show that the statistical significance of two previously proposed gene conversion events can easily be tested and point out that a variety of methods are available to perform gene conversion analyses. PMID- 11734910 TI - Preliminary data on biodistribution and dosimetry for therapy planning of somatostatin receptor positive tumours: comparison of (86)Y-DOTATOC and (111)In DTPA-octreotide. AB - The somatostatin analogue (90)Y-DOTATOC (yttrium-90 DOTA- D-Phe(1)-Tyr(3) octreotide) is used for treatment of patients with neuroendocrine tumours. Accurate pretherapeutic dosimetry would allow for individual planning of the optimal therapeutic strategy. In this study, the biodistribution and resulting dosimetric calculation for therapeutic exposure of critical organs and tumour masses based on the positron emission tomography (PET) tracer (86)Y-DOTATOC, which is chemically identical to the therapeutic agent, were compared with results based on the tracer commonly used for somatostatin receptor scintigraphy, (111)In-DTPA-octreotide (indium-111 DTPA- D-Phe(1)-octreotide, OctreoScan). Three patients with metastatic carcinoid tumours were investigated. Dynamic and static PET studies with 77-186 MBq (86)Y-DOTATOC were performed up to 48 h after injection. Serum and urinary activity were measured simultaneously. Within 1 week, but not sooner than 5 days, patients were re-investigated by conventional scintigraphy with (111)In-DTPA-octreotide (110-187 MBq) using an equivalent protocol. Based on the regional tissue uptake kinetics, residence times were calculated and doses for potential therapy with (90)Y-DOTATOC were estimated. Serum kinetics and urinary excretion of both tracers showed no relevant differences. Estimated liver doses were similar for both tracers. Dose estimation for organs with the highest level of radiation exposure, the kidneys and spleen, showed differences of 10.5%-20.1% depending on the tracer. The largest discrepancies in dose estimation, ranging from 23.1% to 85.9%, were found in tumour masses. Furthermore, there was a wide inter-subject variability in the organ kinetics. Residence times (tau(organs)) for (90)Y-DOTATOC therapy were: tau(liver) 1.59-2.79 h; tau(spleen) 0.07-1.68 h; and tau(kidneys) 0.55-2.46 h (based on (86)Y-DOTATOC). These data suggest that dosimetry based on (86)Y DOTATOC and (111)In-DTPA-octreotide yields similar organ doses, whereas there are relevant differences in estimated tumour doses. Individual pretherapeutic dosimetry for (90)Y-DOTATOC therapy appears necessary considering the large differences in organ doses between individual patients. If possible, the dosimetry should be performed with the chemically identical tracer (86)Y-DOTATOC. PMID- 11734911 TI - Biokinetics and imaging with the somatostatin receptor PET radioligand (68)Ga DOTATOC: preliminary data. AB - Somatostatin (SMS) scintigraphy is widely used for the detection and staging of neuroendocrine tumours. Because of its superior imaging properties, there is growing interest in the use of positron emission tomography (PET) technology for SMS scintigraphy. This study addressed the production of gallium-68 DOTATOC, its biokinetics and its clinical performance in detecting SMS-positive tumours and metastases. A preparation protocol was developed, yielding 40% overall incorporation of (68)Ga into the peptide (DOTATOC). After column filtration, the radiochemical purity exceeded 98%. Eight patients with histologically verified carcinoid tumours were injected with 80-250 MBq of this tracer. PET acquisition was initiated immediately after administration and carried out until 3 h post injection. Images were quantitated using standardised uptake values and target to non-target ratios. Prior to (68)Ga-DOTATOC PET, all patients underwent indium-111 octreotide planar and single-photon emission tomographic (SPET) imaging. Arterial activity elimination was bi-exponential, with half-lives of 2.0 (+/-0.3) min and 48 (+/-7) min. No radioactive metabolites were detected within 4 h in serum. Maximal tumour activity accumulation was reached 70+/-20 min post injection. Kidney uptake averaged <50% compared with spleen uptake. Of 40 lesions predefined by computed tomography and/or magnetic resonance imaging, (68)Ga-DOTATOC PET identified 100%, whereas (111)In-octreotide planar and SPET imaging identified only 85%. Tumour to non-tumour ratios ranged from >3:1 for liver ((111)In octreotide: 1.5:1) to 100:1 for CNS ((111)In-octreotide: 10:1). With (68)Ga DOTATOC >30% additional lesions were detected. It is concluded that PET using (68)Ga-DOTATOC results in high tumour to non-tumour contrast and low kidney accumulation and yields higher detection rates as compared with (111)In octreotide scintigraphy. PMID- 11734912 TI - The impact of FDG-PET on the management algorithm for recurrent colorectal cancer. AB - Positron emission tomography (PET) has been successfully used to image colorectal cancer (CRC). This study evaluated the accuracy of 2-[(18)F]-fluoro-2-deoxy- D glucose (FDG) PET for the detection and staging of recurrent CRC and the consequent impact on clinical management. Forty-two patients previously treated for CRC were investigated for suspected recurrence and, if recurrence was confirmed, the extent of disease was evaluated. All patients underwent whole-body FDG-PET and computed tomography (CT) scan and results were compared to assess sensitivity, specificity and diagnostic accuracy for each modality. We then assessed the FDG-PET directed alteration in clinical management from that planned on the basis of spiral CT results. FDG-PET was more sensitive (93%) than CT (73%) for detection of recurrence (specificity 58% and 75%, respectively). FDG-PET yielded a correct diagnosis in 35 (83%) out of 42 patients, while CT did so in 31 patients (74%). FDG-PET was more accurate than CT for staging local recurrence (sensitivity 100%, specificity 86% with FDG-PET vs 75% and 100%, respectively, with CT) and CRC liver metastases (sensitivity 100% vs 45%; specificity 100% for both). Overall, PET upstaged 8 out of 30 patients (27%) and altered patient management in 16 (38%) cases. This study confirms that FDG-PET is more sensitive than CT for the detection and staging of recurrent CRC. The results also indicate that FDG-PET is an accurate means of selecting appropriate patients for operative treatment. When applied to routine clinical practice, patient management is altered. PMID- 11734913 TI - Evaluation of (pre-)malignant colonic abnormalities: endoscopic validation of FDG PET findings. AB - The diagnostic accuracy of 2-[(18)F]-fluoro-2-deoxy- D-glucose positron emission tomography (FDG-PET) for the detection of (pre-)malignant lesions of the colon was compared with that of endoscopy. We selected a cohort of 39 patients [13 females and 26 males; mean age 62.3 years standard deviation (SD) 9.6 years] who underwent both FDG-PET and endoscopy (total of 44 procedures) in a 2-year period with a maximum interval between the examinations of 3 months (mean 30 days, SD 28 days). The underlying pathology was colorectal malignancies (24 patients), other malignancies (nine patients) and other disorders (six patients). Follow-up of resected colorectal cancer was the most common reason for the performance of endoscopy. In 19 patients FDG bowel uptake was interpreted as non-physiological, and in 18 patients abnormal findings (adenomatous polyps >3 mm or carcinoma) were detected by endoscopy. Compared with colonoscopy, FDG-PET had a sensitivity of 74% and specificity of 84%. The positive predictive value of FDG-PET was 78%. FDG PET failed to detect small (diameter 3-10 mm) polyps in four patients. In nine cases abnormal FDG accumulation on PET imaging was the sole reason for performance of an endoscopic procedure. In these cases, endoscopy detected large adenomatous polyps in four patients and carcinomas in two patients, but no abnormalities were detected on endoscopy in the other three patients. There was a good correlation between the location of FDG uptake and endoscopy-positive lesions. FDG-PET is able to detect clinically relevant lesions of the colon. Our study suggests that it can be regarded as a useful adjunct in the non-invasive follow-up of patients with colorectal carcinomas. PMID- 11734914 TI - Positron emission tomography of hepatic first-pass metabolism of ammonia in pig. AB - Hepatic first-pass metabolism plays a key role in metabolic regulation and drug metabolism. Metabolic processes can be quantified in vivo by positron emission tomography scanning (PET). We wished to develop a PET technique to measure hepatic first-pass metabolism of ammonia. Seven anaesthetised pigs were given positron-labelled ammonia, (13)NH(3), into the portal vein and into the vena cava as successive 2-min infusions followed by 22-min dynamic liver scanning. Vena cava infusion data were used to account for recirculation of tracer and metabolites following the portal vein infusion. The scan data were analysed by a model of sinusoidal zonation of ammonia metabolism with periportal urea formation and perivenous formation of glutamine. The hepatic extraction fraction of (13)NH(3) was 0.73+/-0.16 (mean+/-SD, n=7 pigs). Values of clearance of ammonia to urea and to glutamine were obtained, as were rate constants for washout of these two metabolites. Overall, the modelling showed half of the ammonia uptake to be converted to urea and half to glutamine. The washout rate constant for glutamine was about one-tenth of that for urea. We conclude that hepatic first pass metabolism of ammonia was successfully assessed by PET. PMID- 11734915 TI - Accuracy of technetium-99m sestamibi scintimammography and X-ray mammography in premenopausal women with suspected breast cancer. AB - The aim of this study was to evaluate the usefulness of technetium-99m sestamibi scintimammography (SSM) and combined use of SSM and X-ray mammography (MG) in premenopausal patients with small (< or =2 cm) suspicious breast lesions. Eighty seven premenopausal women (median age 47 years, range 32-52) with breast lesions ranging from 4 to 20 mm in greatest diameter (median 12 mm) and detected by MG, underwent SSM prior to open breast biopsy. A planar camera and single-photon emission tomography (SPET) technique were used in 23 (26.4%) and 64 (73.6%) patients, respectively. At histological examination, 72 (82.8%) breast cancers (pT1a=3, pT1b=27, pT1c=42) and 15 (17.2%) benign masses were found. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy of MG and SSM were 80.6% vs 80.6%, 60.0% vs 93.3% (P<0.05), 90.6% vs 98.3%, 39.1% vs 50.0% and 77.0% vs 82.8%, respectively. MG + SSM together had a sensitivity of 94.4% (P<0.05 vs SSM alone) and an accuracy of 82.8% for the detection of breast cancer. Tumours undetected by both MG and SSM were significantly (P<0.05) smaller than those correctly diagnosed. Patients with false-negative MG were younger (P<0.05) than those with tumours correctly detected, while SSM sensitivity was independent (P=NS) of age. In conclusion, in premenopausal patients with small breast lesions, the sensitivity of SSM and MG is similar (P=NS), whereas the specificity of SSM is significantly higher (P<0.05) than that of MG. Furthermore, the combination of MG and SSM offers significantly improved sensitivity and accuracy in comparison with the individual techniques, suggesting a complementary role of SSM in the detection of breast cancer in younger patients. PMID- 11734916 TI - (99)mTc-tetrofosmin SPET in the detection of both primary breast cancer and axillary lymph node metastasis. AB - The aim of this study was to evaluate the usefulness of (99m)Tc-tetrofosmin single-photon emission tomography (SPET) in the detection of both primary breast cancer and axillary lymph node metastasis. We studied 192 consecutive patients in whom primary breast cancer was suspected on the basis of mammography and/or physical examination. After intravenous injection of 740 MBq (99m)Tc-tetrofosmin, both planar and SPET scintimammography was performed in all patients using a rectangular dual-head gamma camera equipped with low-energy, high-resolution, parallel-hole collimators. In 175 patients with breast cancer at histology, the per-lesion overall sensitivity of SPET and planar imaging for the detection of breast cancer was 95.8% and 75.9% (P<0.0005), respectively. The sensitivity of SPET and planar imaging was, respectively, 96.5% and 79.5% in palpable (P<0.0005) and 90% and 45% in non-palpable lesions (P<0.01). With regard to lesion size, the sensitivity of SPET and planar imaging was, respectively, 90.5% and 45.2% in lesions < or =10 mm ( P<0.0005), 95.3% and 81.4% in lesions of 11-20 mm (P<0.005), 100% and 84.6% in lesions of 21-30 mm (P<0.05) and 100% and 95.8% in lesions >30 mm (P>0.05). In the remaining 17 patients with benign mammary lesions at histology, per-lesion overall specificity of SPET and planar imaging was 76.2% and 85.7% (P>0.05), respectively. Neither SPET nor planar imaging showed false positive results in non-palpable lesions or in those < or =10 mm. In 173 breast cancer patients submitted to axillary lymph node dissection (ALND), per-axilla overall sensitivity of SPET and planar imaging in the detection of axillary lymph node metastasis was 93% and 52.3% ( P<0.0005), respectively. The sensitivity of SPET and planar imaging was, respectively, 100% and 82.6% in palpable nodes (P>0.05), 90.5% and 41.3% in non-palpable nodes (P<0.0005), 92.8% and 35.7% in the presence of < or =3 nodes ( P<0.0005) and 93.2% and 68.2% in the presence of >3 nodes (P<0.005). The specificity of SPET and planar imaging was 91% and 100% (P<0.05), respectively. (99m)Tc-tetrofosmin SPET appears to be a reliable method for the detection of both primary BC and axillary lymph node metastasis, and its diagnostic accuracy exceeds that of (99m)Tc-tetrofosmin planar scintimammography. The use of SPET is particularly important in the identification of small non palpable primary carcinomas and metastatic axillae with < or =3 non-palpable lymph nodes. More extensive use of SPET appears warranted in the management of breast cancer patients. PMID- 11734917 TI - Quantitative analysis of inhomogeneity in ventilation SPET. AB - The aim of this study was to evaluate a method for quantification of inhomogeneity in ventilation single-photon emission tomography (SPET). Nine emphysematous patients, nine life-long non-smokers and nine smokers were included in the study. The SPET investigation was performed after 50 MBq (99m)Tc-Technegas had been inhaled by each subject in the supine position. A single-head gamma camera, equipped with a general-purpose parallel-hole collimator using 64 projections (20 s each) over 360 degrees, was used. Data were acquired in 128x128 matrices. Attenuation correction was applied based upon computed tomography (CT) density maps. Lung regions of interest were delineated manually on CT images and then positioned on SPET images. Several attenuation-corrected transaxial SPET slices (thickness 1 cm, spacing 3.5 cm) were reconstructed. Each SPET slice was divided into several 2x2x1 cm(3) elements. Inhomogeneity was assessed by the coefficient of variation (CV) of the pixel counts within these elements (micro level) and the CV of the total counts of the elements (macro-level). Micro-level CVs in non-smokers varied between 1% and 41%, whereas they were dispersed over a wide range (1%-600%) in emphysematous patients. In seven smokers, the frequency distribution of micro-level CVs was within the normal range, whereas in the other two smokers the values were between the normal range and the range in emphysematous patients. The pooled mean values of micro-level CVs and macro-level CVs in each subject clearly separated the patients from the others. Parametric images of micro-level CV indicated the localisation and severity of ventilation inhomogeneity. We conclude that the present method enables quantification and localisation of regional inhomogeneity in ventilation SPET images. PMID- 11734918 TI - Ventilation-perfusion lung scan for the detection of pulmonary involvement in Takayasu's arteritis. AB - The aim of study was to analyse ventilation and perfusion (V/Q) lung scan findings in a series of Italian patients with Takayasu's arteritis. Eighteen consecutive patients underwent V/Q lung planar scintigraphy and single-photon emission tomography (SPET). Before perfusion scan acquisition was started, a first-pass study with (99m)Tc-macroaggregates of albumin was performed to assess the right ventricular ejection fraction (RVEF). All patients had normal chest X rays and were symptom free at the time of the investigation. They also underwent echocardiography to evaluate pulmonary artery pressure and in 13 patients respiratory function tests were performed. In four patients, perfusion lung scan was repeated after 1 year. In 10/18 patients (55.5%), 43 unmatched lobar, segmental or subsegmental perfusion defects were found on planar images; ventilation scintigraphy was normal in all cases. On SPET images, 55 defects were found; no defects were found with SPET in the remaining patients who had normal planar images. All patients had normal RVEF and 5/13 patients had mild restrictive-obstructive lung disease. The pulmonary artery pressure was increased in two patients with perfusion defects. In the four patients who had repeat scintigraphy, all defects remained unchanged. The prevalence of lung perfusion abnormalities observed in Italian patients with Takayasu's arteritis is within the range of values reported in other countries, and V/Q planar scintigraphy is sufficient for the screening of patients. PMID- 11734919 TI - Early detection of myocardial ischaemia in the emergency department by rest or exercise (99m)Tc tracer myocardial SPET in patients with chest pain and non diagnostic ECG. AB - Chest pain (CP) represents a frequent reason for presentation at the emergency department (ED). A large proportion of patients have non-diagnostic ECG on presentation, and in many cases several hours have elapsed since onset of symptoms. Acute rest myocardial scintigraphy (rest SPET) has been shown to have a relevant role in the detection of patients at risk for coronary events, but its sensitivity and negative predictive value are optimal only within the first 3 h following onset of symptoms. In those with delayed presentation, exercise SPET alone, as a screening approach, appears more promising, but its feasibility and diagnostic role in the ED are still unresolved. A total of 231 consecutive patients with a recent-onset (<24 h) first episode of CP had a negative first line work-up including ECG, troponins, creatine kinase-MB and echocardiography. These patients were considered at low risk for short-term coronary events. Patients were studied with rest SPET if they presented <3 h after onset of CP and exercise SPET if they presented after > or =3 h. The end-points of the study were detection of significant coronary artery disease (CAD) by angiography and major coronary events or cardiac death at 6 months. Eighty patients (35%) underwent rest SPET, while 151 (65%) underwent exercise SPET. Two of the 159 patients with negative SPET had evidence of critical CAD at 6-month follow-up (one patient in the rest SPET group and one in the exercise SPET group; P=NS). Of the 72 patients (31%) with a positive scan, 34 (15%) had documented CAD (16 patients in the rest SPET group and 18 in the exercise SPET group; P=NS). Sensitivity, specificity, accuracy and predictive value were not statistically different between the two groups. In conclusion, the accuracy of exercise SPET in patients with CP and delayed presentation to the ED is comparable to that of validated rest SPET in patients with early presentation. Owing to the high negative predictive value (99%), exercise SPET is especially valuable as a screening tool for the exclusion of CAD in low-risk patients and implementation of early discharge. PMID- 11734920 TI - Relationship between evaluation by quantitative fatty acid myocardial scintigraphy and response to beta-blockade therapy in patients with dilated cardiomyopathy. AB - Predicting the effect of beta-blockade therapy on the clinical outcome of patients with dilated cardiomyopathy (DCM) is difficult prior to the initiation of therapy. Myocardial fatty acid metabolism has been shown to be impaired in patients with DCM. We examined whether the extent of myocardial injury, as assessed by iodine-123 15-( p-iodophenyl)-3- R, S-methylpentadecanoic acid (BMIPP) myocardial scintigraphy, is related to the response of patients with DCM to beta-blockade therapy. Thirty-seven patients with DCM were examined using BMIPP myocardial scintigraphy before and after 6 months of treatment with metoprolol. Myocardial BMIPP uptake (%BM uptake) was estimated quantitatively as a percentage of the total injected count ratio. The left ventricular end diastolic and end-systolic dimensions (LVDd, LVDs) and ejection fraction (LVEF) were also evaluated. The patients were divided into two groups according to their functional improvement (>10% elevation of LVEF) after 6 months of metoprolol therapy. Twenty-eight patients responded to the therapy, while nine did not. Prior to the therapy, no significant differences in LVDd, LVDs or LVEF were observed between the responders and non-responders. However, the %BM uptake was significantly lower in the non-responders than in the responders (1.0%+/-0.2% vs 2.1%+/-0.5%, P<0.001). The %BM uptake could be used to distinguish the responders from the non-responders with a sensitivity of 0.93 and a specificity of 1.00 at a threshold value of 1.4. After the metoprolol therapy, the %BM uptake improved significantly in the responders (2.5%+/-0.5%, P<0.01) but did not change in the non-responders. These results indicate that myocardial BMIPP uptake could predict the response of DCM patients to beta-blockade therapy. PMID- 11734921 TI - Technetium-99m tetrofosmin for parathyroid scintigraphy: a direct comparison with (99m)Tc-MIBI, (201)Tl, MRI and US. AB - The aim of this study was to evaluate the efficacy and role of technetium-99m tetrofosmin for the detection of abnormal parathyroid glands to be referred for surgical treatment. Twenty-eight consecutive patients, including 25 primary and 3 secondary cases of hyperparathyroidism, were evaluated. (99m)Tc tetrofosmin/(99m)Tc-pertechnetate subtraction scintigraphy (TF/Tc) was performed on all patients, and the results were directly compared with those of (99m)Tc methoxyisobutylisonitrile (MIBI)/(99m)Tc-pertechnetate subtraction scintigraphy (MIBI/Tc), (201)Tl/(99m)Tc-pertechnetate subtraction scintigraphy (Tl/Tc), magnetic resonance imaging (MRI) and ultrasonography (US). In cases of single gland disease, the sensitivities of TF/Tc, MIBI/Tc, Tl/Tc, MRI and US were 63.2%, 68.4%, 57.9%, 55.6% and 63.2%, respectively. In cases of multi-gland disease, the sensitivities of TF/Tc, MIBI/Tc, Tl/Tc, MRI and US were 41.7%, 41.7%, 37.5%, 58.3% and 54.2%, respectively. In cases of parathyroid adenoma, the sensitivities of TF/Tc, MIBI/Tc, Tl/Tc, MRI and US were 68.8%, 75.0%, 68.8%, 62.5% and 75.0%, respectively. In cases of parathyroid hyperplasia, the sensitivities of TF/Tc, MIBI/Tc, Tl/Tc, MRI and US were 40.7%, 40.7%, 33.3%, 53.8% and 48.1%, respectively. It is concluded that, for the detection of abnormal parathyroid glands, (99m)Tc-tetrofosmin is as useful as (99m)Tc-MIBI and is more useful than (201)Tl. PMID- 11734922 TI - Cerebral perfusion and haemodynamics measured by SPET in symptom-free patients with transient ischaemic attack: clinical implications. AB - Transient ischaemic attacks (TIAs) are heterogeneous from the clinical, physiopathological, aetiological and prognostic points of view. Single-photon emission tomography (SPET) may influence patient management by helping to define the vascular topography and by suggesting the probable mechanism (embolic or haemodynamic). However, the variables predicting focal regional cerebral blood flow (rCBF) and cerebrovascular reserve (CVR) abnormalities on SPET and their clinical correlation are poorly known. Our objective in this study was to assess the value of rCBF and CVR measured by SPET in a prospective series of 42 patients with recent (within the preceding 30 days) first-ever TIA Two SPET examinations [baseline and post-acetazolamide (ACZ)] were consecutively performed, and region/reference ratios were obtained using an irregular region of interest (ROI) method. Percentages of interhemispheric asymmetry between homologous brain regions were used to identify abnormalities on baseline SPET, and the percentage changes in asymmetry between the baseline and post-ACZ SPET studies were used to identify abnormal responses to the vasodilator stimulus. Mean baseline and test retest values previously obtained in normals were used as a reference. The relationship of SPET findings with clinical data and results of complementary examinations was assessed. Most patients (98%) had abnormal findings on either baseline (43%), post-ACZ (19%) or both SPET studies (36%). Thus, 33 patients had hypoperfusion on baseline SPET (78.5%, symptom related in 45%). In 23 patients, a poor response to ACZ was found (55%, symptom related in 21.5%). No predictors for rCBF or CVR impairment were found. Most patients with a first-ever TIA episode present focal hypoperfusion on SPET, either with or without correlation with TIA symptoms. Post-ACZ SPET increases the probability of finding cerebrovascular abnormalities, and orients attention towards an haemodynamic compromise. Focal hypoperfusion on SPET should not be viewed as clinically insignificant because it probably reflects previous or ongoing clinical and/or subclinical episodes of cerebral ischaemia. PMID- 11734923 TI - 180 degree pinhole SPET with a tilted detector and OS-EM reconstruction: phantom studies and potential clinical applications. AB - This study investigated the feasibility of ordered subsets expectation maximisation (OS-EM) reconstruction of pinhole single-photon emission tomography (SPET) acquired with a tilted detector head and a 180 degrees orbit. Phantom and patient data were recorded using a standard single-head camera. Reconstructions were performed using a dedicated OS-EM algorithm. Reconstructed images of line, uniformity and Picker's thyroid phantoms showed that the geometry, physical size and uniformity of the radioactive objects were preserved. For the range of radius corresponding to the patient studies, the measured full-widths at half-maximum lay between 4.90+/-0.25 mm and 6.05+/-0.25 mm. Finally, the gain in resolution associated with the use of the pinhole collimator instead of a parallel-hole collimator was highlighted in a parathyroid exploration and in a shoulder bone study. PMID- 11734924 TI - The incidence of left iliac fossa uptake of (99m)Tc-DTPA in renal scanning. AB - In rare cases, extrarenal uptake of technetium-99m diethylenetriamine penta acetic acid ((99m)Tc-DTPA) has been reported, e.g. in tumours or abscesses. Although in our experience a relatively common occurrence, increased uptake in the left iliac fossa (LIF) and its incidence have not been described. This retrospective study set out to establish the incidence of this phenomenon and to exclude malignancy as the cause. All patients who had a renal (99m)Tc-DTPA scan between 1 January 1996 and 31 December 1997 had their scans reviewed. Presence and grade of increase in iliac fossa activity were determined by consensus agreement of three observers. The list of patients with increased uptake was cross-referenced against the New South Wales (Australia) Cancer Council database to exclude the possibility of a neoplastic lesion in this region. Increased LIF uptake was noted in 41 of the 231 (18%) consecutive patients (18 men, 23 women; mean age 43 years, range 20-83). Among these 41 patients, uptake was severe in 2 (5%), moderate in 12 (29%) and mild in 27 (66%). No patient had increased uptake in the right iliac fossa. Only one patient had a malignant lesion but this was excluded as being the cause of LIF uptake. No other patient developed malignancy (mean follow-up time 4.1 years; range 3.2-5.1). Increase in LIF uptake is a common, benign finding most likely due to activity within the descending colon. It occurs in approximately 18% of the population, and it is important to recognise such uptake in order to avoid misdiagnoses. PMID- 11734925 TI - Carbon-11 choline or FDG-PET for staging of oesophageal cancer? AB - We investigated the feasibility of using carbon-11 choline (CHOL) positron emission tomography (PET) for the staging of oesophageal cancer, in comparison with fluorine-18 fluorodeoxyglucose (FDG) PET, using histopathological findings as the gold standard. Eighteen patients were studied: 16 patients with cancer of the oesophagus or gastro-oesophageal junction and two with in situ carcinoma/high grade dysplasia. PET imaging was performed 5 min (CHOL) or 90 min (FDG) after injection of 370 MBq of the tracer. PET images were analysed by two independent and blinded physicians using visual and standardised uptake value (SUV) analysis. PET results were compared with surgical and histopathological findings. FDG-PET was able to detect all (100%) of the 16 malignant primary lesions, while CHOL-PET detected 73%. In situ carcinoma ( n=1) and high-grade dysplasia ( n=1) were not visualised with either tracer. Diffuse uptake of the tracers was noted in areas of Barrett's oesophagitis. Twelve patients had locoregional metastases (N1) that were not detected with either FDG or CHOL. Six patients had additional distant nodal (M1a) metastases; four of six (66%) were visualised by FDG, and three of five (60%) by CHOL-PET. On a lesion basis, FDG-PET detected 10/12 non-regional metastases (sensitivity 83%), while CHOL-PET detected 5/12 (sensitivity 42%). Haematogenous distant metastases (M1b) were positive on FDG-PET in three of four patients, and on CHOL-PET in two of four. SUV values were significantly higher for FDG (FDG 6.6+/-3.5, CHOL 5.5+/-2.5, P=0.04). CHOL-PET is able to visualise oesophageal carcinoma and its metastases, but appears to be inferior to FDG-PET. Presumably this is the result of lower tumoural uptake and considerable non specific uptake of CHOL in liver, stomach wall, pancreas and small intestine. Further studies are needed to confirm these data. PMID- 11734927 TI - Anatomical and biochemical investigation of primary brain tumours. AB - Cancerous transformation entails major biochemical changes including modifications of the energy metabolism of the cell, e.g. utilisation of glucose and other substrates, protein synthesis, and expression of receptors and antigens. Tumour growth also leads to heterogeneity in blood flow owing to focal necrosis, angiogenesis and metabolic demands, as well as disruption of transport mechanisms of substrates across cell membranes and other physiological boundaries such as the blood-brain barrier. All these biochemical, histological and anatomical changes can be assessed with emission tomography, X-ray computed tomography (CT), magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS). Whereas anatomical imaging is aimed at the diagnosis of brain tumours, biochemical imaging is better suited for tissue characterisation. The identification of a tumoural mass and the assessment of its size and vascularisation are best achieved with X-ray CT and MRI, while biochemical imaging can provide additional information that is crucial for tumour classification, differential diagnosis and follow-up. As the assessment of variables such as water content, appearance of cystic lesions and location of the tumour are largely irrelevant for tissue characterisation, a number of probes have been employed for the assessment of the biochemical features of tumours. Since biochemical changes may be related to the growth rate of cancer cells, they can be thought of as markers of tumour cell proliferation. Biochemical imaging with radionuclides of processes that occur at a cellular level provides information that complements findings obtained by anatomical imaging aimed at depicting structural, vascular and histological changes. This review focusses on the clinical application of anatomical brain imaging and biochemical assessment with positron emission tomography, single-photon emission tomography and MRS in the diagnosis of primary brain tumours, as well as in follow-up. PMID- 11734930 TI - Contrast-medium-induced nephrotoxicity: are all answers in for acetylcysteine? PMID- 11734931 TI - Paediatric ultrasound. I. Abdominal. AB - Paediatric abdominal US differs partially from adult US. Not only different size and different approaches due to different physiological conditions demand a different handling and equipment, but different diseases and different patient handling characterise the specific challenges of paediatric abdominal US. The main goal of this paper is to familiarize the radiologist who occasionally has to deal with paediatric patients with the specifics and typical applications of US in neonates, infants and children. This review tries to point out the major differences between adult and paediatric scanning, starting from technical requirements and other general considerations and spanning to sophisticated adaptation of modern sonographic imaging techniques to specific paediatric queries. This article illustrates typical applications of the various sonographic approaches and lists some major disease conditions where the role of US has changed and shows how US now significantly contributes to diagnosis. PMID- 11734932 TI - Paediatric ultrasound. II. Other applications. AB - This article focuses on small-parts US in children. The differential diagnosis of cystic neck masses primarily includes lymphangiomas, thyroglossal duct cysts and branchial cleft cysts. Solid masses may be represented from lymphadenopathies, cervical extensions of mediastinal thymus, fibromatosis colli, rhabdomyosarcomas and neuroblastomas. Salivary gland lesions are uncommon in children. Thyroid is best evaluated by US and nuclear scintigraphy. If US shows abnormal thyroid gland, the isotopic scan may be a good complementary method to confirm the diagnosis. Normal parathyroid glands are not visualised routinely by US because of their small size. Parathyroid adenomas are unusual in paediatrics. Ultrasound can be successfully used in the differential diagnosis of the painful scrotum especially with colour flow Doppler. The hallmark of ischaemia is a completely avascular testis. In the paediatric age group, the most common application of US to the musculoskeletal system is the evaluation of the infant hip in the first 6 months of life; however, the refinement of new transducers has further improved the ability of US equipment to evaluate a variety of other musculoskeletal disorders in children, involving tendons, muscles, nerves as well as soft tissue masses. Ultrasound can be a useful screening tool in newborns suspected of having closed spinal dysraphism. PMID- 11734933 TI - Advanced cranial ultrasound: transfontanellar Doppler imaging in neonates. AB - We know that many cerebral lesions are of circulatory origin and it is now important to study cerebral hemodynamics by pulsed and color Doppler ultrasonography. The revolution in the imaging of vascular physiology, the diagnosis, and the prognostic evaluation of vascular disease are not based on morphological sonographic studies but on the Doppler techniques that can display cerebral vessels in the neonate. The results of the hemodynamic investigation in 491 newborns aged from 32 weeks of gestation to 9 months by means of pulsed and color Doppler are reported. Normal values of the resistive index, peak systolic, end-diastolic, and time-averaged velocities in seven different vessels are determined. Some pathological examples are presented. Doppler techniques play a major role in the diagnosis, follow-up, and management of brain damage, whether ischemo-hemorrhagic, infectious, or developmental or tumoral, and of pericerebral collections. PMID- 11734934 TI - Ultrasound of thyroid, parathyroid glands and neck lymph nodes. AB - In the past 15 years high-frequency B-mode sonography and colour-power Doppler have become the most important and most widely employed imaging modalities for the study of the neck, in particular for thyroid gland, parathyroids and lymph nodes. Sonography allows not only the detection but often also the characterization of the diseases of these organs, distinguishing benign from malignant lesions with high sensitivity and specificity, which could be further improved by the employ of ultrasound contrast agents and harmonic imaging. Although no single sonographic criterion is specific for benign or malignant nature of the lesions, the combination of different signs can be markedly helpful to speed up the diagnostic process. Fine-needle aspiration biopsy (FNAB) remains the most accurate modality for the definitive assessment of thyroid gland nodules and of any doubtful case of nodal disease. In association with clinical findings and serum levels of parathormone, FNAB has specificity close to 100% for the characterization of parathyroid adenomas. A combined approach with sonography and FNAB is generally highly effective. PMID- 11734935 TI - Towards a more sophisticated use of breast ultrasound. AB - The diagnostic role of breast ultrasound has been expanded along with the improvement of high-frequency transducers and digital technology. Vascular assessment has progressed enough to depict normal vascular anatomy of the breast and the lymph nodes. Pathologic vessels are seen in almost all the tumors, thus improving US sensitivity for nonpalpable carcinomas. New contrast agents will recirculate enough to search for vascular foci during a thorough investigation of both breasts and nodal stations. The US role in screening might be now revised. Many factors are now in favor of targeted US screening in dense and complex breasts and in high-risk patients. Screening sensitivity is significantly increased. Most of these US-detected tumors are small enough to be curable. Mammography and sonography together are a unique problem-solving and cost effective tool. They can easily guide fine aspirations or larger biopsies reducing the cost of unnecessary surgical procedures. Accurate US investigations facilitate the surgical approach to a very conservative and cosmetic operation. High-resolution sonography can demonstrate the intraductal spread of tumors and their multiple foci more easily than mammography, but US diagnosis is less sensitive than magnetic resonance mammography in the evaluation of the real tumoral extent. Ductal branching has a complex pattern; therefore, intraductal spread and multifocal nodes are better demonstrated by multiplanar analysis of 3D ultrasound data volumes. Sonography can easily explore the different nodal chains. Metastatic disease is indicated by an enlarged and round shape and the absence of the echogenic hilum. Irregularities in the cortex are a very useful sign in metastatic nodes without total replacement of lymphoid tissue by neoplastic cells. These signs are very specific. A time-consuming, radiation emitting and costly sentinel biopsy may be avoided in one of every five clinically node-negative patients. But preoperative US assessment is also important as sonography is very sensitive in patients with extensive nodal involvement that might result negative at the sentinel node procedure. New technologies and contrast agents allow perfusional studies that enhance the contrast resolution and will increase the sensitivity of US for small nodal metastases. PMID- 11734936 TI - Changes in lung parenchyma after acute respiratory distress syndrome (ARDS): assessment with high-resolution computed tomography. AB - The aim of this study was to evaluate the appearance, extent, and distribution of parenchymal changes in the lung after acute respiratory distress syndrome (ARDS) as a function of disease severity and therapeutic procedures. High-resolution computed tomography (HRCT), clinical examination, and lung function tests were performed in 15 patients, 6-10 months after ARDS. The appearance and extent of parenchymal changes were compared with the severity of ARDS, as well as with clinical and therapeutic data. Lung parenchymal changes resembling those found in the presence of pulmonary fibrosis were observed in 13 of 15 patients (87%). The changes were significantly more frequent and more pronounced in the ventral than in the dorsal portions of the lung ( p<0.01). A significant correlation was observed between the extent of lung alterations and the severity of ARDS ( p<0.01), and the duration in which patients had received mechanical ventilation either with a peak inspiratory pressure greater than 30 mmHg ( p<0.05), or with more than 70% oxygen ( p<0.01). Acute respiratory distress syndrome frequently is followed by fibrotic changes in lung parenchyma. The predominantly ventral distribution of these changes indicates that they may be caused by the ventilation regimen and the oxygen therapy rather than by the ARDS. PMID- 11734937 TI - Role of contrast-enhanced helical CT in the evaluation of acute thoracic aortic injuries after blunt chest trauma. AB - The purpose of this retrospective study was to determine the value of contrast enhanced helical CT for detecting and managing acute thoracic aortic injury (ATAI). Between June 1995 and February 2000, 1419 consecutive chest CT examinations were performed in the setting of major blunt trauma. The following CT findings were considered indicative of ATAI: intimal flap; pseudoaneurysm; contour irregularity; lumen abnormality; and extravasation of contrast material. On the basis of these direct findings no further diagnostic investigations were performed. Isolated mediastinal hematoma on CT scans was considered an indirect sign of ATAI: In these cases, thoracic aortography was performed even if CT indicated normal aorta. Seventy-seven patients had abnormal CT scans: Among the 23 patients with direct CT signs, acute thoracic aortic injuries was confirmed at thoracotomy in 21. Two false-positive cases were observed. The 54 remaining patients had isolated mediastinal hematoma without aortic injuries at CT and corresponding negative angiograms. The 1342 patients with negative CT scans were included in the 8-month follow-up program and did not show any adverse sequela based on clinical and radiographic criteria. Contrast-enhanced helical CT has a critical role in the exclusion of thoracic aortic injuries in patient with major blunt chest trauma and prevents unnecessary thoracic aortography. Direct CT signs of ATAI do not require further diagnostic investigations to confirm the diagnosis: Isolated aortic bands or contour vessel abnormalities should be first considered as possible artifacts or related to non-traumatic etiologies especially when mediastinal hematoma is absent. In cases of isolated mediastinal hematoma other possible sources of bleeding should be considered before directing patients to thoracic aortography. PMID- 11734938 TI - Accessory fissures of the lung: evaluation by high-resolution computed tomography. AB - This study was performed to classify and assess the frequency of accessory fissures of the lung by high-resolution CT (HRCT) scans. The HRCT scans of 443 patients were retrospectively reviewed. After exclusion of patients with pulmonary parenchymal distortion, pleural disease or those who had had lobectomy, 186 patients were included in the study. The HRCT scans consisted of 1.5-mm sections obtained at 10-mm intervals and reconstructed with a high-spatial resolution algorithm. In 59 of 186 patients (32%), a total of 69 accessory fissures were detected. The most common fissure was the inferior accessory fissure ( n=40; 21%, 38 on the right, 2 on the left) followed by the left minor fissure ( n=17; 9%). Other accessory fissures observed were the right superior accessory fissure ( n=2; 1%), the azygos fissure ( n=1; 0.5%) and intersegmental fissures between the medial and lateral segments of the right middle lobe ( n=3; 2%), the superior and inferior segments of the lingula ( n=2; 1%), the anterobasal and laterobasal segments of both the right ( n=1; 0.5%) and the left ( n=3; 2%) lower lobe. In conclusion, the inferior accessory fissure and the left minor fissure were the most common accessory fissures seen on HRCT examinations. PMID- 11734939 TI - Tumour detection rate of a new commercially available computer-aided detection system. AB - The aim of this study was to determine the tumour detection rate and false positive rate of a new mammographic computer-aided detection system (CAD) in order to assess its clinical usefulness. The craniocaudal and oblique images of 150 suspicious mammograms from 150 patients that were histologically proven to be malignant were analysed using the Second Look CAD (CADx Medical Systems, Quebec, Canada). Cases were selected randomly using the clinic's internal tumour case sampler. Correct marking of the malignant lesion in at least one view was scored as a true positive. Marks not at the location of the malignant lesion were scored as false positives. In addition, mammograms with histologically proven benign masses ( n=50) and microcalcifications ( n=50), as well as 100 non-suspicious mammograms, were scanned in order to determine the value of false-positive marks per image. The 150 mammograms included 94 lesions that were suspicious due to masses, 26 due to microcalcifications and 30 showed both signs of malignancy. The overall sensitivity was 90.0% (135 of 150). Sensitivity on subsets of the data was 88.7% (110 of 124) for suspicious masses (MA) and 98.2% (55 of 56) for microcalcifications. Eight of 14 false-negative cases were large lesions. The overall false-positive rate was observed as 0.28 and 0.97 marks per image of microcalcifications and masses, respectively. The lowest false-positive rates for microcalcifications and MA were observed in the cancer subgroup, whereas the highest false-positive rates were scored in the benign but mammographically suspicious subgroups, respectively. The new CAD system shows a high tumour detection rate, with approximately 1.3 false positive marks per image. These results suggest that this system might be clinically useful as a second reader of mammograms. The system performance was particularly useful for detecting microcalcifications. PMID- 11734940 TI - First experience with a new dedicated ultrasound system for computer-guided large core breast biopsy. AB - We describe a new dedicated ultrasound system for computer-guided large core breast biopsy (LCBB) and report our first clinical experience in 45 female patients. After an initial 30 biopsy procedures on a US breast phantom, LCBB using this new system was performed in 45 non-palpable surgically verified breast lesions. All biopsies were performed by the same radiologist using 14-gauge long throw biopsy needles. Histological results following LCBB were compared with open surgical biopsy. Procedure time and any complication arising was registered in all procedures. Biopsies using this prototype were successful and yielded sufficient material in all 45 lesions. There were 23 benign and 22 malignant lesions with complete histological agreement between LCBB and open surgical biopsy in 44 of the 45 lesions. In one invasive ductal cancer the pathologist could only state high probability of malignancy but not give a definite diagnosis. Besides one case with early termination due to decreased visibility of the target lesion, no technical complications were noted. Slight vasovagal reactions were seen in 4 patients but did not alter the histological results. The average procedure time was 30+/-2.7 min. This new dedicated US system for computer-guided LCBB is an accurate and safe method for diagnosing breast lesions. Although this new system may have no major impact for US-guided LCBB experienced physicians, it might be a promising alternative for the non-skilled physician to currently available breast biopsy techniques. PMID- 11734941 TI - Calcified pilomatrixoma of the breast: mammographic and sonographic findings. AB - The authors report the ultrasonographic and mammographic features of a case of pilomatrixoma of the breast, a rare benign neoplasm originating from the hair matrix. PMID- 11734942 TI - Bilateral breast involvement in Sezary syndrome. AB - Cutaneous T-cell lymphoma is a term used for mycosis fungoides and Sezary syndrome, the distinct clinical entities where the skin is the primary organ of involvement. Sezary syndrome is the leukemic variant of mycosis fungoides, presenting with generalized erythroderma, lymphadenopathy, and atypical cells (the Sezary cells) in the peripheral blood and bone marrow. The dissemination of cutaneous T-cell lymphoma may occur with no exception of the organs; however, no prior report exists about the Sezary syndrome secondarily involving the breasts. We report the clinical and radiological findings of bilateral breast involvement in a case of Sezary syndrome. PMID- 11734943 TI - Warthin's tumor of parotid gland on Tc-99m pertechnetate scintigraphy with lemon juice stimulation: Tc-99m uptake, size, and pathologic correlation. AB - The aim of this study was to evaluate the usefulness of technetium-99m (Tc-99m) pertechnetate scintigraphy with lemon juice stimulation in the diagnosis of Warthin's tumor and its correlation with Tc-99m uptake, tumor size, and histologic subtype. Tc-99m pertechnetate scintigraphy before and after lemon juice stimulation and pathologic specimens of 34 Warthin's tumors and 47 non Warthin's lesions were retrospectively evaluated. Tc-99m uptake of Warthin's tumors before and after stimulation was visually graded as follows: absent; indeterminate; low grade; definite; and strong. Tumor size was defined as maximum diameter of the tumor measured from the surgical specimen. Warthin's tumors were classified into three histologic subtypes according to the ratio of epithelial and lymphoid stromal components: predominant epithelial; intermediate; and low grade epithelial types. Eighteen of 34 (53%) Warthin's tumors and one benign lymphoepithelial cyst showed higher uptake than that of the normal parotid gland on Tc-99m scintigraphy before lemon juice stimulation. Thirty-two of the 34 (94%) Warthin's tumors, one benign lymphoepithelial cyst, one pleomorphic adenoma, and one oncocytoma revealed higher uptake than that of the normal parotid gland on Tc 99m scintigraphy after lemon juice stimulation. The mean size was 37 mm in strong uptake Warthin's tumors, 24 mm in definite uptake tumors, 19 mm in low-grade uptake tumors, and 12 mm in low-grade uptake tumors excluding those tumors with large cystic component. There was a significant correlation between tumor size and degree of Tc-99m uptake after lemon juice stimulation. However, there was no correlation between histologic subtype and Tc-99m uptake, and histologic subtype and tumor size in Warthin's tumors. Our study concludes that Tc-99m pertechnetate scintigraphy with lemon juice stimulation is useful for the detection and diagnosis of Warthin's tumor. The degree of uptake in Warthin's tumor on Tc-99m scintigraphy with lemon juice stimulation depends mainly on tumor size and the presence of large cystic component in it. PMID- 11734944 TI - Laryngeal squamous cell carcinoma presenting as a prelaryngeal neck abscess: report of two cases. AB - Head and neck tumours presenting as a neck abscess are extremely rare. Two patients with laryngeal squamous cell carcinoma, presenting with a prelaryngeal abscess, are described. Although clinically and on the CT studies these cases were suspect for underlying malignancy, the initial biopsies were negative for cancer. Because of persistent suspicion, repeat biopsies finally confirmed the presence of a squamous cell carcinoma. Pathological examination after total laryngectomy showed cancer localised in one of the true vocal cords, invading the anterior commissure and thyroid cartilage. However, the prelaryngeal soft tissues were free of tumour, showing only inflammatory changes and collections of pus. These cases emphasize the importance of repeat targeted biopsies and follow-up CT studies in patients with a prelaryngeal abscess of obscure aetiology, in order to exclude or confirm a possible underlying malignancy. PMID- 11734945 TI - Adrenal hemorrhage after orthotopic liver transplantation: MR appearance. AB - The purpose of this paper is to describe the MR imaging findings of right adrenal hemorrhage after orthotopic liver transplantation. Twenty-seven orthotopic liver transplantation patients underwent MR studies of the liver and/or biliary system. Patients were referred to MR examination because of suspected biliary complications ( n=22) or for evaluation of mass lesions ( n=5). The standard MR protocol included T1-weighted spin-echo (SE) or gradient-recalled echo (GRE) images and T2-weighted turbo SE (TSE) images with fat suppression. In addition, cholangiography pulse sequences and/or contrast-enhanced T1-weighted images were obtained according to specific indications. In 2 patients a right adrenal mass was detected at MR imaging. Three to 4 weeks after transplantation, the lesions were markedly hyperintense on T2-weighted images and showed a hypointense capsule. Follow-up MR examinations revealed a slight decrease in size and a change in morphology. Computed tomography examinations of these 2 patients, obtained 10 weeks after transplantation, showed resolution of the hemorrhage and transformation into a cystic lesion in one case and a complete resolution of the hemorrhage and a normal right adrenal gland in the other case. Adrenal hemorrhage after liver transplantation shows typical MR features and should not be mistaken for an adrenal tumor or a postoperative abscess. PMID- 11734946 TI - Contrast-enhanced MR angiography in patients after kidney transplantation. AB - The aim of this study was to investigate the value of a contrast-enhanced 3D MR angiography in detecting postoperative vascular complications after kidney transplantation in comparison with digital subtraction angiography (DSA). Forty one patients who underwent a kidney transplantation were examined with MR angiography and DSA. Contrast-enhanced MR angiography was performed as a dynamic measurement with one precontrast and three postcontrast measurements. Maximum intensity projection reconstructions were performed for all postcontrast data sets after DSA. The results were evaluated by two independent observers who were unaware of the DSA results. Twenty-three hemodynamically significant arterial stenoses were identified with DSA in the iliac arteries ( n=7), the renal allograft arteries ( n=12), and in their first branches ( n=4). For a patient based analysis the sensitivity and specificity, respectively, for observer 1 were 100 and 97%, and for observer 2, 100 and 93%. Respective data were 100 and 100% after a consensus evaluation by two observers. Complications involving the renal veins were detected in 2 cases and perfusion defects of the kidney parenchyma were detected in 4 cases. Contrast-enhanced MR angiography is a reliable method in identifying postoperative arterial stenoses after kidney transplantation. In addition, dynamic MR angiography can be helpful in detecting venous complications and perfusion defects in kidney allografts. PMID- 11734947 TI - Contrast-enhanced power Doppler US in the diagnosis of renal pseudotumors. AB - The term "pseudotumor" is used to refer to several anatomic variants that can simulate a renal mass, the most frequent of which are hypertrophied column of Bertin, persistence of fetal lobation, and the dromedary or splenic hump. We describe the findings of power Doppler US after the ultrasound contrast agent (Levovist, Schering, Berlin, Germany) administration in 4 patients with a renal focal lesion in whom gray-scale and baseline power Doppler US was not able to certainly differentiate pseudotumor from neoplasm. PMID- 11734948 TI - Seminal vesicle cyst associated with ipsilateral renal agenesis and papillary carcinoma of the bladder. AB - We report a case of seminal vesicle cyst associated with ipsilateral renal agenesis in a 37-year-old patient with papillary carcinoma of the bladder. Ultrasonography showed absence of the right kidney, a bladder tumour and a round retrovesical hypoechogenic mass with posterior acoustic enhancement. It showed low attenuation on CT, low signal intensity on T1-weighted MR images and high signal intensity on T2-weighted MR images. Intravenous urography (IVU) and cystoscopy were also performed. After surgery, it proved to be a seminal vesicle cyst. The embryology, imaging characteristics and differential diagnosis of seminal vesicle cysts are discussed. Associated findings are also described. PMID- 11734949 TI - Multiphase contrast-enhanced helical CT of liver metastases from renal cell carcinoma. AB - The aim of this study was to evaluate whether in patients with metastatic renal cell carcinoma (RCC) multiphase liver studies would improve detection of metastatic liver disease. Forty-six consecutive patients with known metastatic RCC underwent standardized non-contrast and triphasic contrast enhanced hepatic CT examinations as part of their routine imaging studies. Once a liver abnormality was detected, it was characterized as metastatic by a panel of three radiologists who followed pre-set criteria. These criteria included change in size, biopsy results and lack of benign features. Presence and conspicuity of liver metastases were graded using a five-point scale by consensus of a panel of three radiologists. The highest number of lesions evaluated per patient was limited to ten. Seventy-two liver metastases were detected in 16 patients. Of these, 54 were seen on unenhanced scans; 47 in the hepatic arterial (HA) phase, at 25 s; 65 in the portal-venous (PV) phase, at 60 s; and 49 in delayed images, at 90 s. Scanning only during the PV phase would have missed seven lesions (10%), six of which were seen on unenhanced images and six were seen in HA phase. All patients with metastatic liver disease would have been identified by combination of unenhanced and PV phase or by HA and PV phase scanning. Forty-two lesions were graded more conspicuous on the PV phase, whereas 18 (25%) were more conspicuous on the HA phase. The combination of unenhanced, HA and PV scanning should be considered in the initial evaluation of patients with metastatic RCC for improved lesion detection and characterization. Subsequently, the combination of unenhanced and PV phase imaging is preferred. PMID- 11734950 TI - Atypically enhancing hepatic cavernous hemangiomas: high-spatial-resolution gadolinium-enhanced triphasic dynamic gradient-recalled-echo imaging findings. AB - We retrospectively investigated the appearance and frequency of atypically enhancing cavernous hemangiomas with high-spatial-resolution (512x224 matrix) gadolinium-enhanced triphasic dynamic gradient-recalled-echo (GRE) MR images. Images of 132 hepatic cavernous hemangiomas (ranging in size from 4 to 72 mm; mean size 17.2 mm) in 95 patients (42 men and 53 women; age range 25-85 years; mean age 54 years) were retrospectively reviewed by two independent radiologists. Forty (30%) of 132 lesions atypically enhanced. Smaller hemangiomas (< or =15 mm) more frequently (29%) showed early entire enhancement with or without arterio portal shunting in the hepatic arterial-dominant phase ( p<0.001); most of them showed hyperintense complete fill-in in the equilibrium phase and were readily characterized. "Bright dot" or minimal peripheral enhancement in the equilibrium phase was seen in a small number of lesions (6% each). With T2-weighted images, 130 (98%) lesions showed moderately to very high signal intensity and only 2 (2%) with minimal peripheral enhancement showed hyperintensity of slight degree. The high-spatial-resolution dynamic GRE images clearly revealed minute enhancement characteristics of hemangiomas. Although moderately to very high signal intensity with T2-weighted MR images is informative for the diagnosis of most cavernous hemangiomas, when a lesion shows minimal peripheral enhancement in the equilibrium phase and hyperintensity of slight degree with T2-weighted images, further follow-up or biopsy may be warranted to discriminate hypovascular metastases. PMID- 11734951 TI - MR cholangiography in the diagnosis of sclerosing cholangitis in Langerhans' cell histiocytosis. AB - Langerhans' cell histiocytosis (LCH) is a disorder of histiocytic proliferation that primarily affects infants. Imaging findings of a rare case of lung and liver involvement in an adult are presented. High-resolution computed tomography (HRCT) of the lungs showed confluent thin-walled cystic air spaces compatible with advanced LCH. Liver CT and MRI revealed unspecific signs of fatty infiltration. Irregular widening of peripheral bile ducts was displayed in breath-hold MR cholangiography. This pattern is considered characteristic for sclerosing cholangitis and should support the diagnosis of LCH in case of concomitant cystic pulmonary disease, even in adult patients. PMID- 11734952 TI - Direct communication between the parabiliary and paraumbilical venous network: a case report. AB - A case with a direct communication between the parabiliary vein and the paraumbilical vein observed on CT obtained after portal vein embolization is presented. The object of this paper is to draw attention to this rare venous communication since it can serve as various collateral pathways. PMID- 11734953 TI - Percutaneous transluminal renal angioplasty in atheroma with renal failure: long term outcomes in 99 patients. AB - The aim of this study was to evaluate renal function changes after percutaneous transluminal renal artery angioplasty (PTRA) done to treat atheromatous renal artery stenosis with renal failure. Between 1990 and 1995, PTRA was performed in 99 renal failure patients (creatinine clearance less than 80 ml/min) with atheromatous stenosis of one or more native renal arteries. Indications for PTRA were chronic renal failure with poorly controlled hypertension (group A, 67 patients) or rapidly deteriorating renal function (group B, 32 patients). Renal function changes after PTRA were evaluated based on the percentages of patients with improved, stabilized, and worsened serum creatinine and creatinine clearance values, and on mean differences between final and baseline creatinine clearances. At the end of follow-up (19+/-10 months), group A had a significantly smaller creatinine clearance gain (42.9 ml/min before PTA to 44.5 ml/min after PTA, D=1.6 ml/min, in group A, vs 24.1-28.4 ml/min, D=4.3, in group B, p=0.03), and a significantly smaller percentage of improved patients (36 vs 62%) than group B. Most stenoses in group B either were bilateral or occurred on a solitary kidney ( p=0.001). Percutaneous transluminal renal artery angioplasty combined with aggressive medical treatment may be useful in maintaining or improving renal function, particularly in patients with a recent, sharp deterioration in renal function. PMID- 11734954 TI - In vivo evaluation of intravascular stents at three-dimensional MR angiography. AB - The aim of this study was to assess in vivo patency of seven commonly used non ferromagnetic plain stents as regards demonstration of the contained lumen at 3D gadolinium-enhanced MRA in a 0.5-T MR environment. Twenty-one patients with 22 stents (1 aortic, 18 iliac, 3 popliteal) were imaged with MRA. Stent-related artefacts were evaluated. Images were assessed for the presence and size of stent artefacts, luminal patency, and were correlated with stent metal mass, composition, configuration and angulation with respect to the magnetic field. Four different types of stent (Symphony, Wallstent, Memotherm, SMART) showed major artefacts preventing assessment of intradevice luminal patency or restenosis, in two stents (ZAV stent, Intracoil) minor artefacts allowed assessment of high-degree stenosis, and only one type of stent (Strecker) was perfectly imaged. Based on our limited experience, Gd-MRA appears well suited for the evaluation of only a minority of MR-compatible stents. PMID- 11734955 TI - Low molecular weight heparin and the risk of haemorrhage following percutaneous biopsy, despite a normal standard clotting screen. AB - There has been an increase of the use of low molecular weight heparin in the treatment of thrombotic events. This case report describes a complication of a pelvic mass biopsy performed whilst the patient was being treated with low molecular weight heparin (LMWH). Despite an uncomplicated biopsy procedure and confirmation of normal clotting screen, INR (International normalised ratio), APTR (Activated partial thromboplastin ratio) and platelet levels, the biopsy was complicated by severe haemorrhage. PMID- 11734956 TI - Abdominal aortic aneurysm in Behcet's disease: treatment with endovascular stent graft. AB - A case of acute rupture of an abdominal aortic aneurysm in a patient with Behcet's disease is reported. The patient was successfully treated by implantation of an endovascular stent graft. The preinterventional diagnostic procedures and the postinterventional follow-up are described and the benefit and risk vs open surgery is discussed. PMID- 11734957 TI - CT-guided injection of botulinic toxin for percutaneous therapy of piriformis muscle syndrome with preliminary MRI results about denervative process. AB - Piriformis muscle syndrome (PMS) is a cause of sciatica, leg or buttock pain and disability. The pain is usually increased by muscular contraction, palpation or prolonged sitting. The aim of our paper was to evaluate the feasibility of CT guided percutaneous botulinic toxin (BTX) injection for the purpose of PMS treatment. Thirty patients suffering from PMS, suspected with clinical and electrophysiological criteria, after imaging examinations excluding other causes of sciatic pain, resulted positive at the lidocaine test and were treated by intramuscular injection of BTX type A under CT guidance. The follow-up (12 months) was performed with clinical examination in all cases and with MR 3 months after the procedure in 9 patients to evaluate the denervative process entity of the treated muscle. In 26 cases relief of symptoms was obtained after 5-7 days. In 4 patients an insufficient relief of pain justified a second percutaneous treatment which was clinically successful. No complications or side effects were recorded after BTX injection. The MR examination showed a signal intensity change of the treated muscle in 7 patients due to the denervative process of PM, whereas in the remaining 2 cases only an atrophy of the treated muscle was detected. Larger series are necessary to confirm these MRI preliminary results. The CT guided BTX injection in the PMS is an emergent and feasible technique that obtains an excellent local therapeutic effect without risk of imprecise inoculation. PMID- 11734958 TI - MR imaging of synovial tumors and tumor-like lesions. AB - The clinical diagnosis of synovial tumors and tumorlike lesions is difficult, and radiographic findings may establish a confident diagnosis only in some cases. MR imaging has become the modality of choice in evaluating these lesions, because with it a presumptive diagnosis can be made in most cases. Our goal is to review the MR features of pigmented villonodular synovitis, giant-cell tumor of the tendon sheath, synovial chondromatosis, synovial hemangioma, lipoma arborescens, synovial cysts and synovial sarcoma, emphasizing those findings that suggest a specific diagnosis. PMID- 11734959 TI - Levator claviculae muscle presenting as a hard clavicular mass: imaging study. AB - We report a case of levator claviculae muscle presenting clinically as a hard mass in the clavicular area due to angular deformity of this bone. To our knowledge, this is the first report where the anomalous muscle shows this clinical presentation. PMID- 11734960 TI - High-resolution CT of paediatric lung disease. AB - High-resolution computed tomography (HRCT) has improved our understanding of many lung diseases in adults. The technique is used less often in children due to concerns regarding radiation dose. However, HRCT may provide important diagnostic information in a variety of lung diseases in children including airways diseases and diffuse interstitial lung disease. This review illustrates the HRCT appearances of a variety of conditions and describes the emerging role of the technique in children. PMID- 11734961 TI - Sonographic features indicative of hydrostatic reducibility of intestinal intussusception in infancy and early childhood. AB - The aim of the study was to investigate the various sonographic patterns of intussusception, which may be indicative of its reducibility by hydrostatic reduction. Seventy-one infants and children clinically suspected of having intussusception were studied sonographically and given barium enemas. Sonographic patterns of intussusception were correlated with its reducibility. When the head of intussusception appeared as a target-like mass the hydrostatic reduction rate was 100%. When demonstrated as a doughnut-like mass, the intussusception reducibility depended on the thickness of the hypoechoic external ring of the "doughnut". With a thickness equal to, or less than, 7.2 mm the reduction rate was 100%, with a thickness ranged between 7.5 and 11.2 mm the reduction rate was 68.9% and with a thickness measured 14 to 14.2 mm a surgical resection of bowel was required. When a small amount of fluid appeared within the head of intussusception, like a crescent hypoechoic area, the hydrostatic reduction was unsuccessful, regardless of the sonographic pattern of intussusception (target- or doughnut-like mass). The presence of a small amount of free peritoneal fluid did not affect the reducibility of intussusception. In conclusion, sonographic patterns of intussusception identify good and poor prognostic features for the success of hydrostatic reduction. PMID- 11734962 TI - Respiratory syncytial virus infection of the lower respiratory tract: radiological findings in 108 children. AB - For years the typical appearance of respiratory syncytial virus (RSV)-induced infection of the lower respiratory tract has been discussed. All available studies have led to different results. The aim of this study was to control these results, with 108 children. The age range was 1 day to 10 years (median 7 months). Within 72 h of admission, all children developed an RSV infection of the lower respiratory tract. Chest X-rays (pa-view) of 55 children under, and 53 children over, the age of 6 months (10/53>24 months) were evaluated. The diagnosis of RSV and the chest X-ray were mostly done on the same day. The major radiological findings of the two age-groups were compared by Wilcoxon's unpaired rank sum test. Major radiological findings were: normal chest X-ray (30%), central pneumonia (32%) or peribronchitis (26%). There was no statistical significance between the age-groups. Other findings were emphysema (11%), pleural effusion (6%), lobar- or broncho-pneumonia (each 6%), atelectasis (5%) or pneumothorax in one case. Therefore, the most common radiological findings in RSV induced infection of the lower respiratory tract, supported by our results (RSV infection without bacterial superinfection) are central pneumonia, peribronchitis or normal chest X-ray. Thus an age-group separation into under or over 6 months is no longer necessary. PMID- 11734963 TI - Transient neonatal hypoglycemia: cranial US and MRI findings. AB - A case of transient neonatal hypoglycemia with patchy hyperechogenic white matter abnormalities in the frontal and parietooccipital lobes on cranial US is presented. An MRI examination revealed T1 and T2 shortening of the lesions in the occipital and frontal white matter. Follow-up cranial US demonstrated recovery of white matter changes in the patient with normal neurological outcome. PMID- 11734964 TI - Prenatal testicular torsion: sonographic appearance in the newborn infant. AB - The prenatal diagnosis of spermatic cord torsion is often really difficult and the diagnosis is usually retrospective. Herein, we report a case of a male newborn baby who presented at delivery with an enlarged, swollen and tender scrotum. US showed an enlarged right testis, with dishomogeneous texture, fluid collection between the testis and the tunica vaginalis and large hydrocele. Differential diagnosis included hydrocele complicated by infection or hemorrhage, testicular tumor or postnatal testicular torsion. Color and power Doppler did not reveal any flow signal, and the diagnosis of antenatal torsion with initial necrosis was made. The role of color Doppler US is emphasized in directing the patient to emergency surgical exploration, when testicular salvage may be possible. Delayed surgical treatment can be proposed, when the diagnosis of antenatal torsion has a high degree of certainty. However, the Doppler examination of a newborn baby's testis is a very difficult challenge even for an experienced radiologist. PMID- 11734965 TI - Intraocular metastases: differential diagnosis from uveal melanomas with high resolution MRI using a surface coil. AB - Magnetic resonance imaging with dedicated surface coils plays a pivotal role in differential diagnosis and staging of intraocular tumors. The purpose of this study was to establish MRI criteria for the differential diagnosis of uveal melanomas and intraocular metastases. In a prospective study 44 eyes in 36 patients with intraocular metastases and 200 patients with uveal melanomas were investigated with MRI using a 1.5-T scanner and a 5-cm surface coil. Both quantitative and qualitative evaluation of the resulting images was performed. The MR signal intensities typically expected for metastases (slightly hyperintense on non-contrast T1-weighted images and hypointense on T2-weighted images compared to the vitreous body) were seen in only 23.1%. The typical melanoma signal of either moderate or strong hyperintensity on T1-weighted images and hypointensity on T2-weighted images was seen in 69.4% of the proven melanomas. Contrast enhancement was observed in both metastases and melanomas. Morphological differences between metastases and melanomas were detected in tumor size, shape, position, frequency of retinal detachment, and homogeneity of the tumor. Differentiation between intraocular metastases and uveal melanoma is limited by overlap of signal intensities. Some improvement is achieved with morphologic criteria. PMID- 11734966 TI - Diffusion-weighted MR imaging of Carmofur-induced leukoencephalopathy. AB - Carmofur (1-hexylcarbamyl-5-fluorouracil), a derivative of 5-fluorouracil (5-FU), has been widely used in Japan as a postoperative adjuvant chemotherapy agent for colorectal and breast cancer. Periventricular hyperintensity on T2-weighted MR images in carmofur-induced leukoencephalopathy confront the physician with a broad range of differential diagnoses. We describe two cases of carmofur-induced leukoencephalopathy in which diffusion-weighted MR imaging revealed periventricular hyperintensity. We compared their findings with those of age related periventricular hyperintensity in five patients and found discrepancies in signal intensity of periventricular areas. Our results suggest that diffusion weighted MR imaging may be useful to differentiate carmofur-induced leukoencephalopathy from age-related periventricular hyperintensity. PMID- 11734967 TI - Anterior spinal artery stroke demonstrated by echo-planar DWI. AB - Diffusion-weighted MR (DWI) is becoming an established method for the investigation of cerebral ischemia. Its value in spinal ischemia has to be demonstrated. We report on a patient presenting with postoperative paraparesis who underwent emergency MRI of the spine with echo-planar diffusion-weighted imaging which showed an area of hyperintensity corresponding to a decrease of diffusion as measured by the apparent diffusion coefficient. On follow-up imaging spinal stroke was confirmed. In conclusion, spinal echo-planar MR imaging can demonstrate ischemic changes despite strong echo-planar artifacts. It could become an important adjunct to the management of patients with suspected spinal ischemia. PMID- 11734968 TI - Technetium-99m-ECD SPECT in antiphospholipid antibody syndrome: a drastic improvement in brain perfusion by antiplatelet therapy. AB - We present a case of antiphospholipid antibody syndrome (APS) with repeated transient ischemic attacks (TIAs). Magnetic resonance imaging showed multiple cerebral infarcts and ischemic changes in the cerebral white matter. Cerebral angiographies showed no abnormalities. Technetium-99m-ethyl cysteinate dimer (Tc 99m-ECD) brain SPECT showed multiple decreased perfusion areas, which were more extensive than the lesions demonstrated on MRI. After treatment with an antiplatelet agent, the patient subsequently recovered from the TIAs. Although no interval changes were observed by MRI after therapy, follow-up Tc-99m-ECD SPECT revealed a marked improvement in brain perfusion. This is the first imaging report of remarkable post-therapy improvement in brain perfusion in APS cases. PMID- 11734969 TI - Developments in digital radiography: an equipment update. AB - Digital X-ray imaging technology has advanced rapidly over the past few years. This review, particularly aimed at those involved in using and purchasing such technology, is an attempt to unravel some of the complexities of this potentially confusing subject. The main groups of X-ray imaging devices that are considered are digitisers of conventional radiographs, image-intensifier-based fluorography systems, photostimulable phosphor computed radiography, amorphous selenium-based technology for thorax imaging and flat-panel systems. As well as describing these different systems, we look at ways of objectively assessing their image quality. Concepts that are used and explained include spatial resolution, grey-scale bit resolution, signal-to-noise ratio and detective quantum efficiency. An understanding of these basic parameters is vital in making a scientific assessment of a system's performance. Image processing and techniques are also briefly discussed, particularly with reference to their potential effects on image quality. This review aims to provide a basic understanding of digital X-ray imaging technology and enables the reader to make an independent and educated assessment of the relative merits of each system. PMID- 11734970 TI - A rational approach to dose reduction in CT: individualized scan protocols. AB - The aim of this study was to demonstrate that dose reduction and constant image quality can be achieved by adjusting X-ray dose to patient size. To establish the relation between patient size, image quality and dose we scanned 19 patients with reduced dose. Image noise was measured. Four radiologists scored image quality subjectively, whereby a higher score meant less image quality. A reference patient diameter was determined for which the dose was just sufficient. Then 22 patients were scanned with the X-ray dose adjusted to their size. Again, image noise was measured and subjective image quality was scored. The dose reduction compared with the standard protocol was calculated. In the first group the measured noise was correlated to the patient diameter (rho=0.78). This correlation is lost in the second group (rho=-0.13). The correlation between patient diameter and subjective image quality scores changes from rho=0.60 (group 1) to rho=-0.69 (group 2). Compared with the standard protocol, the dose was reduced (mean 28%, range 0-76%) in 19 of 22 patients (86%). Dose reduction and constant noise can be achieved when the X-ray dose is adjusted to the patient diameter. With constant image noise the subjective image quality increases with larger patients. PMID- 11734971 TI - Requests for body computed tomography: increasing workload, increasing indications and increasing age. AB - Increasing numbers of increasingly elderly patients were being examined in our Body CT department. At the same time, some of our clinical colleagues perceived that their patients might be discriminated against on the basis of their age when allocating CT time. We therefore studied the population trends in our department over a 10-year period. The ages of patients attending the Body CT department were collected from the hospital's computer information system from 1995 to 2000 and from handwritten logbooks for the months of September 1988 and 1998. Comparison was made with population trends within the hospital and local demographic data. There has been an average increase of 11% per annum in the number of examinations performed in the Body CT unit. The average age of patients examined increased from 52.7 years in 1988 to 58.9 years in 1998. The largest increase occurred in the over 75-year population (18% rise per annum). Hospital and local demographic population profiles changed little during the same period. We are performing increasing numbers of body CT examinations on increasingly elderly patients. This is probably due to an increased willingness to investigate and treat elderly patients, rather than changes in the local population. There is no evidence of a general discriminatory policy on the basis of age. PMID- 11734972 TI - Spontaneous regression of low-grade astrocytomas: an underrecognized condition? PMID- 11734974 TI - Comments on Kalender et al.: a PC program for estimating organ dose and effective dose values in computed tomography. PMID- 11734981 TI - Anthony J. Raimondi ISPN Award. PMID- 11734982 TI - Spondylolysis and spondylolisthesis in the pediatric and adolescent population. AB - This article is a review of spondylolysis and spondylolisthesis in younger age groups. Since Herbinaux first described the pathology (1782), many classifications and theories of etiopathogenesis have been proposed. The congenital and isthmic types, as classified by Wiltse, are the most frequent in younger age groups, but the postsurgical progressive forms (3-5%) have been described as increasing in frequency secondary to neoplastic surgery in children. The general incidence is 4-5% at the age of 6 years, and in 30-50% of cases these types do not progress to spondylolisthesis. Most cases are asymptomatic (80%). Standard radiographic examinations (A-P, L, Oblique) are helpful in diagnosis and can suggest what the prognosis will be in terms of the evolution, and also what treatment is indicated (degree of slippage, slip angle, lumbar and lumbosacral index, SPTI). A bone scan (PBS and SPECT) is useful in the early stages of spondylolysis (pre-spondylosis). Although the CT scan is the most accurate examination, MRI is becoming important for diagnosis because of the frequency with which it is used as a primary investigation method. Depending on patient age, progression, degree of slippage, and symptoms, different therapeutic approaches have been proposed and are described in this paper. PMID- 11734983 TI - Childhood meningioma: unusual location, atypical radiological findings, and favorable treatment outcome. AB - OBJECTS: To investigate the characteristics of childhood meningioma, especially, locations, radiological findings, pathological features (including proliferative potential) and outcome, 11 children with meningiomas were retrospectively analyzed. RESULTS: Unusual location, large size, frequent calcification, and cyst formation were characteristic radiological findings. Gross total resection was achieved in 8 patients, and there was recurrence in 2. Gamma knife radiosurgery was performed on residual and recurrent tumors. MIB-1 indices tended to be high in large tumors. Nine patients had a Karnofsky Performance Scale of more than 70 during the follow-up period of 10 months to 19.5 years. Surgical treatment rendered 4 of 5 epileptic patients seizure free. The childhood meningiomas examined had unusual locations, atypical radiological findings, and various proliferative potentials. CONCLUSIONS: Complete resection is the treatment of choice. Gamma knife radiosurgery can be a good alternative for residual tumors and small recurrent tumors. The outcome of childhood meningiomas is good after surgery. PMID- 11734984 TI - Posthemorrhagic hydrocephalus in preterm infants: long-term follow-up and shunt related complications. AB - INTRODUCTION: A retrospective analysis of 42 preterm infants with intraventricular hemorrhage was performed in order to evaluate shunt-related problems in neonates with posthemorrhagic hydrocephalus. PATIENTS AND METHODS: Within the last 15 years, 76 infants with intraventricular hemorrhage were treated primarily by external ventricular drainage, and 42 needed permanent shunting. Shunting was performed 28-101 days after the patients' birth, ventriculoatrial shunts being placed in 10 patients and ventriculoperitoneal shunts in 32. RESULTS: The mean number of shunt revisions per patient was 1.57. The main reasons for shunt revision were infection (7.1%) and blockage (45.2%). Primary ventriculoperitoneal and ventriculoatrial shunts differed only insignificantly in revision rate and length of shunt survival, whereas ventriculoperitoneal shunts used in shunt revisions required significantly fewer further revisions. CONCLUSIONS: The neurological outcome was not related to the necessity for or to complications of a shunt. Programmable valve systems and neuroendoscopy appear to be helpful in the overall management and in the treatment of complications. PMID- 11734985 TI - The management of immediate post-traumatic seizures in children following minor head injury--time for a multicentre study? AB - BACKGROUND: Children have a much lower threshold for seizure activity than adults. Consequently, an immediate post-traumatic seizure in a child following a minor head injury does not have the same potentially serious implications as one in an adult. METHODS: The records of children admitted with a post-traumatic fit occurring within 24 h of a minor head injury and with a normal CT scan of the brain on admission were critically reviewed. Those with previous neurological disorders, especially fits (excepting febrile convulsions), or systemic injuries requiring admission to an intensive care unit were excluded. RESULTS: There were 13 children who were not intubated and ventilated, forming the control group, and 13 children who were intubated and ventilated and formed the study group. All the children in both these groups made a good recovery, and none of them had any neurological deficit either at discharge or at follow-up. CONCLUSION: The clinical data suggest that children with immediate post-traumatic seizures following a minor head injury whose CT scan shows no major intracranial abnormalities and who have no prior history of neurological disease are at low risk of developing recurrent seizures or neurological complications and can be safely managed without recourse to intubation and ventilation. PMID- 11734986 TI - Atretic cephalocele--report of two cases with special reference to embryology. AB - We report two cases of atretic cephalocele, a diverse form of cranium bifidum. The patients were 15-year-old and 3-month-old girls, who each had a hard, nonpulsatile, nonreducible lump covered by alopecic scalp in the parieto occipital area. They were surgically treated. In case 2, microscopical examination of the operative specimen revealed a meninges under the mass, which was devoid of nervous tissue. Such lesions have rarely been reported, and their essential nature is still the subject of controversy. Pathological and embryological aspects of atretic cephalocele are discussed on the basis of the findings; the neural crest remnant was assumed to be the developmental origin of the lesion in each of these cases. PMID- 11734987 TI - Atretic cephalocele communicating with lateral ventricles. AB - Atretic cephaloceles are thought to be involuted true meningoceles or encephaloceles. They are located at the midline of the parietal or occipital region and usually joined to the subarachnoidal space. We present a 6-month-old girl with atretic cephalocele communicating with the lateral ventricles. PMID- 11734988 TI - Childhood posterior fossa pial-dural arteriovenous fistula treated by endovascular occlusion. AB - Posterior fossa dural arteriovenous fistulae in children are rare lesions that are difficult to treat effectively. We report a case of a 2-year-old girl with such a fistula, who presented clinically with progressive neurological deficit. The lesion was treated by endovascular occlusion, resulting in radiological and clinical cure. PMID- 11734989 TI - Identification of an arteriovenous fistula in a child. Case report and review of the literature. AB - BACKGROUND: A 6-year-old girl sustained a subarachnoid hemorrhage after a mild head injury and was discovered to have an arteriovenous fistula (AVF). INVESTIGATIONS AND TREATMENT: The etiology of subarachnoid hemorrhage was not evident on the initial brain CT. Brain CT with CT angiography identified the lesion. The AVF was further imaged with brain MRI followed by cerebral angiography and successfully embolized. OUTCOME: The child did not suffer any neurological sequelae. PMID- 11734990 TI - Diastematomyelia associated with ectopic dysplastic renal tissue--report of a rare case. AB - BACKGROUND: Presence of heterotopic dysplastic renal tissue in the lumbosacral region is an extremely uncommon condition. CASE REPORT: We report the first case of diastematomyelia associated with ectopic renal tissue. A 10-month-old male child presented with lipomeningomyelocele associated with spina bifida in the lumbosacral region, and the lipoma was excised. Imaging of the spine at 5 years of age showed spina bifida, bony diastematomyelia, lipomeningomyelocele and a small intraspinal cystic lesion. The boy was then operated upon at the age of 5 years, and histopathological examination of the cystic lesion revealed ectopic dysplastic renal tissue. PMID- 11734996 TI - Overexpression of cyclin A in human HeLa cells induces detachment of kinetochores and spindle pole/centrosome overproduction. AB - The combination of hydroxyurea (HU) and caffeine has been used for inducing kinetochore dissociation from mitotic chromosomes and for causing centrosome/spindle pole amplification. However, these effects on microtubule organizing centers (MTOCs) are limited to certain cell types. It was reasoned that if the biochemical differences in MTOC behavior between cells following HU treatment could be identified, then critical information concerning the regulation of these organelles would be obtained. During these studies, it was determined that cells from hamster, rat, and deer could be induced to enter mitosis with dissociated kinetochores and to synthesize centrosomes during arrest with HU, while cells from human and mouse could not. Comparisons between human HeLa cells and CHO cells determined that cyclin A levels were depressed in HeLa cells relative to CHO cells following HU addition. Overexpression of cyclin A in HeLa cells converted them to a cell type capable of detaching kinetochores from mitotic chromosomes. Ultrastructural analyses determined that the detached human kinetochores exhibited a normal plate-like morphology and appeared capable of associating with microtubules. In addition, HeLa cells overexpressing cyclin A also overproduced spindle poles during HU arrest, demonstrating that cyclin A activity also is important for centrosome replication during interphase. In summary, elevated cyclin A levels are important for the capacity of cells to be driven into mitosis by caffeine addition, for the ability of cells to progress to mitosis with detached kinetochores, and for centrosome/spindle pole replication. PMID- 11734997 TI - INCENP loss from an inactive centromere correlates with the loss of sister chromatid cohesion. AB - Inactive centromeres of stable dicentric chromosomes provide a unique opportunity to examine the resolution of sister chromatid cohesion in mitosis. Here we show for the first time that inactive centromeres are composed of heterochromatin, as defined by the presence of heterochromatin protein HP1(Hs alpha). We then show that both the inner centromere protein (INCENP) and its binding partner Aurora B/AIM-1 kinase can also be detected at the inactive centromere. Thus, targeting of the chromosomal passengers is not dependent upon the presence of an active centromere/kinetochore. Furthermore, we show that the association of INCENP with the inactive centromere correlates strictly with the state of cohesion between sister chromatids: loss of cohesion is accompanied by loss of detectable INCENP. These results are consistent with recent suggestions that one function of the chromosomal passenger proteins may be to regulate sister chromatid separation in mitosis. PMID- 11734998 TI - Interaction of DNA polymerase beta with GRIP1 during meiosis. AB - DNA polymerase beta (pol beta) is an essential enzyme that has been shown to localize as discrete foci to the synaptonemal complex during meiosis in the mouse. To identify proteins that associate with pol beta during meiosis, we employed the yeast two-hybrid screen. Here we show that a multiple PDZ domain containing protein, the glutamate receptor interacting protein 1 (GRIP1), interacts specifically with pol beta. The PDZ domain-containing proteins, including GRIP1, act as scaffolds to promote rapid and localized biochemical events that require the interaction of multiple proteins. GRIP1 localizes to discrete foci on meiotic bivalents of both spermatocyte and oocyte nuclei, and colocalizes with pol beta. Together, these findings provide evidence that GRIP1 interacts with pol beta during meiosis. Our findings are consistent with the possibility that GRIP1 acts as a scaffold to promote interaction between proteins that function during meiosis. PMID- 11734999 TI - Expression of Xist RNA is sufficient to initiate macrochromatin body formation. AB - MacroH2A1 is a histone variant that is found as a component of the inactive X chromosome where it is detected as a dense accumulation called a macrochromatin body (MCB). Macrochromatin bodies co-localize with Xist RNA, which is an untranslated RNA that is expressed exclusively from the inactive X chromosome of placental mammals. However, no studies to date have investigated whether Xist RNA expression is necessary or sufficient to cause the formation of MCBs. Here we show that expression of Xist RNA is sufficient to cause the formation of MCBs even when Xist is expressed from an inducible transgene at ectopic autosomal sites. Macrochromatin bodies form at sites of transgenic Xist expression in differentiating mouse ES cell lines and transgenic fibroblasts, but MCBs cannot form in undifferentiated ES cells even after prolonged Xist expression. The kinetics of MCB formation revealed that Xist expression precedes MCB formation and that differentiating ES cells undergo a rapid and synchronous transition that renders them competent to form MCBs. Once MCBs have formed, continued expression of Xist is required for their maintenance. These results show that Xist RNA and macroH2A1 function in a common pathway. Expression of Xist in a permissive nuclear environment is sufficient to initiate a chromatin-remodeling event culminating in the incorporation of macroH2A1. The results also strongly suggest the existence of additional regulatory factors for X inactivation that are regulated developmentally. In addition, we present evidence that macroH2A1 density is not simply a measure of the general degree of DNA compaction. PMID- 11735000 TI - Neo-centromere formation on a 2.6 Mb mini-chromosome in DT40 cells. AB - We describe a mammalian artificial mini-chromosome lacking human alphoid DNA and mouse minor and major satellite DNA repeats. This mini-chromosome, initially recovered in a mouse embryonic stem (ES) cell line (CGR8), is 2.6 Mb in size and consists of sequences derived from the human Y chromosome and mouse chromosomes 12 and 15. It is not stable in the CGR8 cells but replicates and segregates with high fidelity after transfer into chicken DT40 cells. Combined analysis by immunocytochemistry/fluorescence in situ hybridisation (FISH) on metaphase spreads detected an active neo-centromere on the mini-chromosome in these cells. Further analysis by immunocytochemistry/FISH on stretched chromatin allowed the localisation of the CENP-C protein to the DNA sequence derived from interval 5 of the human Y chromosome. PMID- 11735001 TI - The Drosophila maternal gene sesame is required for sperm chromatin remodeling at fertilization. AB - The spermatozoon features an extremely condensed and inactive nucleus. The unique sperm chromatin organization is acquired during the late stages of spermatid differentiation by the replacement of somatic histones with sperm-specific chromosomal proteins. At fertilization, the inactive sperm nucleus must be rapidly transformed into a DNA replication competent male pronucleus before the formation of the zygote. The sequential events of this crucial process are well conserved among animals and are controlled by molecules present in the egg. We have previously identified a Drosophila maternal effect mutation called sesame, which specifically arrests male pronucleus formation at a late stage of chromatin decondensation. In this study, we show that sesame affects maternal histone incorporation in the male pronucleus, a situation that is expected to prevent nucleosomal organization of the paternal chromatin. As an apparent consequence, the male pronucleus is arrested before the first S-phase and does not condense mitotic chromosomes. However, centromeric heterochromatin is present on paternal centromeres, which occasionally interact with microtubules. The abnormal chromatin organization of the male pronucleus does not prevent the formation of a male pronuclear envelope, which breaks down and reassembles in synchrony with maternally derived nuclei present in the same cytoplasm. PMID- 11735002 TI - Expression of vascular antigens by bone cells during bone regeneration in a membranous bone distraction system. AB - An in vivo system of membranous bone formation during distraction has been investigated in order to follow cells that express vascular markers with the objective of understanding the neovascularization process. Concomitantly, sustained proliferation of preskeletal cells was achieved through the application of mechanical force. New capillaries and leading edges that arose by angiogenesis from the periosteal and mucosal surfaces and invaded the central zone of the regenerating distraction tissue temporally preceded the growth of delicate woven bone trabeculae from both edges of the cut bone. Concentrically arranged 'onion like' configurations were abundant in paracentral zones and in association with mesenchymal condensations, suggesting their de novo formation in situ. Vascular specific markers, the angiopoietin receptor Tie-2 and factor VIII-related antigen (FVIIIrAg), were localized immunohistochemically in order to follow cells of vascular origin. Endothelial cells of the new capillaries, centrally located cells of the concentric configurations, pericytes, and most of the adjacent polygonal mesenchymal cells stained positively with specific antibodies to both antigens. Moreover, preosteoblasts and osteoblasts that lie adjacent to or already embedded in the osteiod of the newly formed trabeculae were also FVIIIrAg and Tie-2 immunopositive. As the source of the bone-forming cells in regenerating tissue during distraction is not yet fully understood, this observation might support the possibility of their vascular origin. PMID- 11735003 TI - Partial identification of the mab (CD)Amp1 antigen at the epithelial-mesenchymal interface in the developing kidney. AB - The nature of the primary functional events of nephron induction is still unknown, making it impossible to completely understand the mechanism of tissue interaction between collecting duct ampulla and the surrounding nephrogenic mesenchyme. Soluble morphogenic substances are known to be exchanged in the process and it is assumed that nephron induction requires close contact between both tissues involved. Contrasting with that assumption our previous investigation revealed a thick fibrous meshwork separating nephron inducer and mesenchyme. Our present investigation focused on the molecular characterization of the mab (CD)Amp1 antigen, which is found only in this meshwork. The protein was shown immunohistochemically to be located exclusively at the embryonic collecting duct ampulla and could be clearly distinguished from other extracellular matrix proteins such as collagen type IV, laminin, reticulin, and fibronectin. Two-dimensional electrophoresis of the soluble form of P(CD)Amp1 showed a molecular weight of 87,000 and an isoelectric point of 4.3-4.4. Results from N-terminal sequencing indicated a partial sequence homology of P(CD)Amp1 to collagen type IV alpha 2-chain precursor but additionally yielded unknown sequences. Thus P(CD)Amp1 is a novel, collagen-related protein, restricted to the fibrous meshwork at the mesenchymal-epithelial interphase, which is the site of primary epithelial-mesenchymal interaction. PMID- 11735004 TI - Molecular polymorphism of the intermediate filament protein transitin. AB - Transitin is an avian intermediate filament protein whose transient expression in the progenitor cells of the muscle and nerve tissues is similar to that of mammalian nestin. Both proteins contain an alpha-helical core domain flanked by a short N-terminal head and a long C-terminal extremity. However, the tail region of transitin is significantly different from that of nestin in that it harbors a unique motif containing more than 50 leucine zipper-like heptad repeats which is not found in any other intermediate filament protein. Despite the absence of introns in this region of the transitin gene, it was reported that different isoforms of the protein were produced by exclusion or inclusion of a number of repeats generated by an unusual splicing mechanism recognizing consensus 5' and 3' splice sites contained within the coding sequence of the heptad repeat domain [Napier et al. (1999) J Mol Neurosci 12:11-22]. Two monoclonal antibodies (mAbs) reacting with repeated epitopes of this motif were used to monitor transitin expression during in vitro myogenesis of the quail myogenic cell line QM7. Confocal microscopy revealed that the subcellular domains decorated with mAbs A2B11 and VAP-5 were mutually exclusive: the intermediate filament network visualized with mAb VAP-5 appeared to abut on a submembranous domain defined by mAb A2B11. When QM7 cells were induced to differentiate by switching to medium containing low serum components, an early effect was the local loss of A2B11 cortical staining at the points of cell-cell contacts. The A2B11 signal also disappeared before that of VAP-5 in newly formed myotubes. Unexpectedly, the mutually exclusive staining pattern of the mAbs could not be explained by alternative splicing since both epitopes mapped to a repeated element preceding the consensus 5' splice sites of the heptad repeat domain. An alternative explanation would be that the central repeat domain of transitin is a polymorphic structure from which different conformations exist depending on the local context. This hypothesis is strengthened by the observation that in cultured neural crest cells, the A2B11 antigen is preferentially expressed by freely migrating crest cells whose intracellular pH and calcium concentrations are different from those of non-migrating cells. PMID- 11735005 TI - Endothelin immunocytochemistry: indications of false-positive labeling patterns and non-detectable antigen concentrations. AB - Endothelin is an endothelium-derived peptide with potent vasoconstrictor and mitogenic properties. Since studies concerning the immunocytochemical localization of endothelin are often inconsistent we tried to clear up some of these discrepancies by comparing specificity and labeling patterns of different endothelin antibodies. Monoclonal and polyclonal endothelin antibodies ( n=7) were examined concerning their reactivity with endothelins and heart tissues by immunoblotting. Using immunofluorescence microscopy reactivities with human non failing hearts, failing hearts, and cultured endothelial cells were examined. Specificity of labelings was assessed by absorption controls and functional controls using endothelial cells conditioned to produce different amounts of endothelin. As shown by immunoblotting five out of seven endothelin antibodies revealed both specific endothelin reactivity and negligible non-specific reactivities with heart proteins. Immunocytochemistry showed vascular reactivity of N-terminal endothelin antibodies to be associated with alpha-smooth muscle actin expression. One N-terminal antibody showed additional nuclear reactivity, and one C-terminal antibody vimentin-like labeling patterns. Although these reactivities were abolished in absorption controls these labelings had to be graded non-specific because functional specificity of endothelin antibodies could not be proven. The remaining antibodies revealed no endothelial reactivity even after tyramide signal amplification. Cellular endothelin concentrations ranged around 2,000-fold below the detection limit of immunocytochemical methods. Discrepancies in endothelin immunocytochemistry may originate from false-positive results and from expression levels of endothelin below the detection limit of immunocytochemical methods. PMID- 11735006 TI - Immunohistochemical localisation of androgen receptor during sex-specific morphogenesis in the fetal mouse. AB - The distribution of androgen receptors (ARs) in paraffin serial sections of day 17 and day 18 male and female mouse embryos was investigated. In the cranial section of the genital tract AR expression was restricted to Wolffian structures while Mullerian ducts and surrounding mesenchyme were AR negative. In the fusion zone with the urogenital sinus the epithelial components of the vaginal bud were clearly distinguished by differential AR expression, which was faint in the Wolffian ducts, totally missing in the Mullerian ducts, and intense in the sinus ridges with the most intense expression in the morphogenetically active mesenchyme, indicating a new mechanism of negative control of vagina formation via androgens. Expression of ARs outside the genital tract was observed: (1) in loose interstitial mesenchyme extending into the retroperitoneal space up to the coeliac artery, indicating androgen effects during ascent of the kidneys and descent of intraperitoneal organs, (2) in the trigone of the bladder indicating androgen involvement in the development of the vesico-ureteral junction, and (3) in loose mesenchyme between striated muscle fibres and around pelvic skeletal elements, indicating mediation of androgen effects on the musculoskeletal system via loose mesenchyme. PMID- 11735007 TI - Immunohistochemical localization of beta 1-adrenergic receptors in the liver of male and female F344 rat. AB - The distribution of beta(1)-adrenergic receptors in the liver of Fischer 344 (F344) rat has been examined by an immunohistochemical method. The study was carried out on formalin-fixed and paraffin-embedded livers from young adult, middle-aged, and old female and male F344 rats. An antibody specific for the beta(1)-adrenoreceptor subtype was used. A positive reaction was found in the liver parenchyma of female and male rats from all age groups. Within the liver lobule, a clear zonation is observed, with the beta(1)-adrenoreceptor positivity most evident in pericentral zone hepatocytes and a gradual fading of the immunostaining from pericentral to periportal zone hepatocytes, which may be completely negative. Immunoreactivity is localized on the cell membrane and on the membrane of peripheral cytoplasmic vesicles, and is mostly confined to the cell side facing vascular space. The intensity of immunostaining seems to be slightly higher in the 6- and 10-month-old female rats as compared to the matched male rats and to the senescent female rats. No age-related changes in the intensity of immunostaining are appreciable in male rats. However, no definite conclusion could be drawn about the existence of gender-related differences or age-related changes in the density of beta(1)-adrenoreceptors. A low density of beta1-adrenoreceptor was observed in the spontaneous preneoplastic lesions of the livers from senescent rats. PMID- 11735008 TI - The localization of the relaxed form of plasminogen activator inhibitor type 2 in human gingival tissues. AB - The plasminogen activating system is important in extracellular proteolysis. Plasmin degrades tissues and activates proteases. Plasminogen activators (tissue type; t-PA and urokinase type; u-PA) and plasminogen activator inhibitors (PAI-1, PAI-2) are found in high concentrations in gingival crevicular fluid (GCF). Previous findings indicate the significance of PAI-2 in gingival inflammation. When PAI-2 inhibits a plasminogen activator its conformation relaxes and neoepitopes can be detected with a monoclonal antibody (#2H5). Our aim was to study if and where in the gingival region PAI-2 has acted as an inhibitor. Methodological studies were performed on GCF with western blotting. Frozen sections of human gingiva were studied immunohistochemically. The methodological studies showed that our antibody #2H5 selectively detects relaxed low molecular weight non-glycosylated PAI-2. Total PAI-2 and relaxed PAI-2 were found in all gingival epithelia with a honeycomb-like staining. Relaxed PAI-2 showed the most pronounced staining in the cell layers near the surface of the epithelium and no staining in the suprabasal layers, while total PAI-2 was found throughout the epithelium, often more pronounced suprabasally. The results showed that PAI-2 indeed has acted as an inhibitor of a protease in gingival tissues, primarily in the epithelia. The results also suggest primarily an intracellular localization and thus the interaction of PAI-2 with a protein other than t-PA. PMID- 11735009 TI - Immunolocalization of hepatocyte growth factor and its receptor (c-Met) during mouse liver development. AB - Although hepatocyte growth factor (HGF) was discovered as a potent hepatotrophic factor responsible for liver regeneration and may involve some organ development in embryogenesis, it remains to be revealed what roles HGF plays in liver development. The present study was undertaken to determine which cells express HGF and its receptor c-Met and when c-Met is activated in mouse liver development by using immunoblotting and immunohistochemical techniques. HGF was detected in hepatocytes and non-parenchymal cells, including biliary epithelial cells, periportal connective tissue cells, megakaryocytes, endothelial cells, and sinusoidal cells, throughout liver development. Positive HGF immunostaining in hepatocytes increased during postnatal development, and reached the maximal level in the adult stage. c-Met protein was also expressed in hepatocytes throughout liver development, but maximal staining was obtained in 1- or 2-week-old livers. Phosphorylation of tyrosine residues in the c-Met beta chain also occurred in these stages. These results suggest that HGF signaling is implicated in hepatocyte growth during postnatal liver development, and its action could be in a paracrine mode; HGF produced by non-parenchymal cells such as sinusoidal cells acts on hepatocytes expressing c-Met receptors. Positive immunostaining in adult and postnatal hepatocytes may be derived from their blood clearance of HGF. PMID- 11735011 TI - Efficacy of oral ofloxacin for single-dose perioperative prophylaxis in general surgery--a controlled randomized clinical study. AB - BACKGROUND: Perioperative antibiotic prophylaxis surely reduces surgical infection rate. Pharmacokinetic data of oral ofloxacin in combination with its antibacterial spectrum suggest effective protection against perioperative infection. In addition, costs, adverse effects, and induction of microbial resistance are low. Therefore we performed a controlled randomized study comparing oral and intravenous single dose prophylaxis. METHODS: A total of 61 patients undergoing colonic or pancreatic resection randomly received either a single dose standard intravenous prophylaxis or ofloxacin 400 mg and metronidazole 500 mg orally 2 h before surgery. Postoperative infections were recorded for 3 weeks. RESULTS: Groups were very well comparable regarding age, overweight, concomitant disease, type and duration of surgery, blood loss, and volume support. Infectious complications occurred in 14.8% after parenteral and 3.3% after enteral antibiotic prophylaxis. There was no difference in post operative hospital stay. CONCLUSION: The data demonstrate that single-dose oral ofloxacin is at least as effective as a standard intravenous prophylaxis in patients with colonic or pancreatic resection. It offers significant advantages regarding costs and ease of administration. PMID- 11735012 TI - Nasogastric tube versus gastrostomy tube for gastric decompression in abdominal surgery: a prospective, randomized trial comparing patients' tube-related inconvenience. AB - BACKGROUND: Perioperative decompression of the stomach is still a common procedure and can be achieved using either nasogastric tubes (NTs) or gastrostomy tubes (GTs). While both procedures appear to be equally effective, some authors believe that NTs are less convenient for patients than GTs. However, to date, no reliable prospective data are available on this issue. METHODS: We conducted a prospective, randomized trial comparing NTs versus GTs with a total of 110 patients undergoing elective colon surgery. The primary outcome measure was the patient's tube-related inconvenience and pain, assessed in a standardized interview on day 2 after surgery and quantified by means of a visual-analog scale (VAS). A questionnaire including the EORTC QLQ-C30 and additional items regarding retrospective tube-related judgements was administered on the day of discharge and 4 weeks after discharge. Secondary endpoints were the therapy-related morbidity and general complications. RESULTS: When patients were asked which of their drainage tubes (all patients had three or four drainage tubes, such as decompression drains, urinary drains, central venous line) was most inconvenient, 43% (CI 33-53%) in the NT group reported that the NT was most inconvenient, while only 4% (CI 1-10%) of the GT patients judged the GT most inconvenient ( P<0.001, Chi(2) test). This effect was also found in VAS ratings of inconvenience and discomfort ( P<0.01). Also postoperatively (p.o.), NT patients evidenced less preference for their tube system (day 2 p.o.: 71%, CI 61-80%; 4 weeks p.o.: 66%, CI 56-75%) than did GT patients (day 2 p.o.: 94%, CI 88-98%; 4 weeks p.o.: 91% CI 84-96%); again, these differences were statistically significant ( P<0.02; Chi(2) test). No differences between groups emerged regarding global quality of life or conventional clinical outcomes. CONCLUSION: This prospective randomized trial supports the clinical observation that NT causes more subjective inconvenience than GT. In cases when a prolonged postoperative ileus is expected, decisions for a prophylactic decompressing tube system have to weigh up the possibilities of different averse clinical as well as subjective outcomes. It is then preferable to include patients' preferences in the individual decision making process (shared-decision making). PMID- 11735013 TI - Gallstone surgery in German university hospitals. Development, complications and changing strategies. AB - A representative anonymous questionnaire was sent to all German university hospitals ( n=45) to address the development and outcome of laparoscopic gallstone surgery between 1991 and 1998. The response rate was 64%, and 28,753 operations for gallstone disease were analysed. Two-thirds of the procedures (67%) were performed laparoscopically. While a significant decrease of surgical complications (1991 vs 1998: 5.4% vs 3.4%; P<0.001) and re-laparotomy rate (1.0% vs 0.5%, P<0.05) was observed, no significant changes were detected with regard to the mortality rate. A learning curve regarding common bile duct (CBD) injuries was detected, showing a significant increase between 1991 and 1994 (0.3% and 0.7%, respectively, P<0.05) and a decrease to 0.2% in 1995 and 1996 ( P<0.05). The use of intraoperative cholangiography is compulsory in 10%, selective in 52%, while 38% of the university hospitals never use it. Most institutions use the Veress needle for pneumoperitoneum installation, and 93% use hook cautery for dissection. Since 1998, 45% of all institutions have also used the harmonic scalpel. The spectrum of indications for laparoscopic procedures has increased with time since acute cholecystitis, CBD stones and adhesions are not considered a general reason for a primary open approach. Our findings confirm a learning curve with regard to postoperative morbidity after laparoscopic cholecystectomy. This observation may be due to better training as well as surgical experience. PMID- 11735014 TI - Standardized axillary lymphadenectomy improves local control but not survival in patients with palpable lymph node metastases of cutaneous malignant melanoma. AB - BACKGROUND: The aim of the present study was to investigate whether consistent application of standard surgical techniques contributes to improved local control or improved survival in melanoma patients with palpable lymph node metastases to the axilla. METHOD: Using Kaplan-Meier estimations and the Cox proportional hazards model, 120 axillary dissections were assessed. The data from 63 patients who had received standardized axillary lymphadenectomy were compared to those of 57 patients operated on before the introduction of standardized lymphadenectomy. RESULTS: The introduction of standardized axillary lymphadenectomy led to a significant decrease in 5-year local recurrence probability from 43.1% to 13.1% ( P=0.0008). Using multifactorial analysis, the standardized operative procedure resulted in a significantly lower risk of recurrence in the dissected axilla ( P=0.009). The development of intransit metastases was also related to recurrence in the nodal basin ( P=0.03). No significance was accorded to the number of metastatic lymph nodes, tumor thickness, epidermal ulceration, site of primary melanoma, age, sex, or adjuvant chemotherapy. Notably, no differences in the survival rates were observed between the two treatment groups. CONCLUSION: In patients with palpably enlarged axillary lymph node metastases, the introduction of a complete lymphadenectomy, standardized according to the currently acknowledged principles of oncological surgery, improves local disease control without improving overall survival. PMID- 11735015 TI - Abdominal aortic aneurysm in liver transplant recipients. AB - BACKGROUND AND AIMS: To analyze the incidence, clinical features, expansion rate of, and clinical approach to abdominal aortic aneurysm in patients who had undergone orthotopic liver transplantation. To our knowledge, this is the first report on this issue in liver transplant recipients. PATIENTS/METHODS: Among 172 patients undergoing 185 liver transplantations at our institution over the last 10-year period, we identified three patients (1.7%) with infrarenal aortic aneurysm. They had all undergone routine pre-liver transplant ultrasonography screening for aortic aneurysm. RESULTS: All three patients were symptom free at the time of the discovery of a mild infrarenal abdominal enlargement before ( n=2) and after liver transplantation ( n=1), and were closely monitored by ultrasonography in the follow-up period (3.1-4.3 years). The mean aneurysm expansion rate was 0.73 cm/year. All patients underwent aneurysm repair after their aneurysm expanded significantly under observation, with a mean diameter of 5.1 cm at the time of repair. All three patients are alive and well (median follow-up: 19 months). CONCLUSIONS: Our data suggest that careful ultrasonographic surveillance is warranted in any liver transplant recipient, because of the apparent propensity for a more rapid aneurysm expansion and potentially aggressive course than in the untransplanted population. Early repair of the infrarenal aneurysm is recommended in transplant recipients, given that excellent perioperative and late outcomes can be achieved. PMID- 11735016 TI - Influence of donor MHC class I antigen expression on graft survival after rat parathyroid allotransplantation. AB - BACKGROUND AND AIMS: We investigated the influence of donor MHC antigen expression on graft survival after parathyroid transplantation in three different strain combinations. METHODS: MHC class I and II expression on parathyroid tissue of Lewis (LEW), Dark Agouti (DA), and Wistar-Furth (WF) rats was first analysed semiquantitatively by immunohistochemistry. Additionally, five groups were transplanted: (1) LEW to LEW, (2) DA to DA, (3) LEW to DA, (4) WF to LEW, and (5) DA to LEW. METHODS: MHC class I expression was strong in DA, moderate in WF, and weak in LEW rats; MHC class II expression was negative in all three strains. In the interstitium of all investigated tissue specimens, the proportion of MHC class II-expressing cells was low. RESULTS: After syngeneic transplantation, graft survival could be documented over the whole observation period. A mean graft survival of 20 (+/-2) days was observed following transplantation from LEW to DA, grafts in the group WF to LEW were rejected after 13 (+/-1) days, and graft function lasted 8 (+/-2) days in the group DA to LEW. The number of intragraft leukocytes expressing MHC class II molecules was equal in all groups, whereas increased levels of MHC class I on rat parathyroid tissue before transplantation resulted in a more rapid rejection. CONCLUSION: These results demonstrate that immunogenicity of rat parathyroid tissue seems to be determined by the amount of MHC class I expressed on donor parenchymal cells. PMID- 11735017 TI - Calcitonin kinetics in the early postoperative period of medullary thyroid carcinoma. AB - INTRODUCTION: Calcitonin is a sensitive marker for medullary thyroid carcinoma. Normalisation of calcitonin levels following resection of medullary thyroid carcinoma has been described after a few hours; however, it may be observed more than 4 weeks after surgery. The aim of this study was to correlate the postoperative calcitonin kinetics with preoperative calcitonin levels and tumour stage. Furthermore, we wanted to test the prognostic impact of the calcitonin kinetics. Therefore, only patients with postoperative normalisation of calcitonin levels (biochemical cure) were included in this study. METHODS: Fourteen biochemically cured patients were analysed, including measurement of postoperative basal and pentagastrin-stimulated calcitonin concentration. With respect to the time of postoperative basal calcitonin normalisation, patients were classified into two groups: (A) patients with normalisation of basal calcitonin levels within 24 h and (B) patients with normalisation of basal calcitonin levels later than 24 h postoperatively. RESULTS: Eight patients were found to have normalisation of basal calcitonin levels within 24 h (group A). In the remaining six patients (group B), the period to normalisation of basal calcitonin levels varied from 6 days to 14 days and longer. There were no differences between the two groups with regard to tumour size, number and pattern of lymph node metastases and tumour stage. However, preoperative basal calcitonin levels were significantly different (258 ng/ml vs 955 ng/ml, P<0.01). In the group with slow-decreasing calcitonin levels, no strong correlation between the preoperative level and the postoperative time to normalisation of basal calcitonin levels could be established, which may be due to the small number of patients. After a median follow-up of 21 months, no patient developed tumour recurrence. However, an increased basal calcitonin level was observed in one patient from group B. All other patients had normal basal and peak calcitonin levels. CONCLUSION: Using a highly sensitive calcitonin assay, we demonstrated that normalisation of basal calcitonin levels may be delayed in patients suffering from medullary thyroid carcinoma. The lack of correlation of preoperative levels and the time to normalisation of the basal calcitonin levels, as well as the positive pentagastrin test in some of the patients, argues that this phenomenon is not simply due to prolonged biochemical calcitonin elimination. Nevertheless, a prognostic influence could not be shown in this study due to the short follow up-period. Further investigations and a longer follow-up are necessary to determine the nature and the prognostic impact of delayed normalisation of calcitonin levels. PMID- 11735018 TI - Intraoperative measurement of intact parathyroid hormone in renal hyperparathyroidism by an inexpensive routine assay. AB - Although the kinetics of intraoperative intact parathyroid hormone (iPTH) are well characterised in primary hyperparathyroidism, no data are available for patients with renal hyperparathyroidism and renal insufficiency, partially because of the high costs of intraoperative quick iPTH measurement. Therefore we evaluated an inexpensive laboratory test with a duration of 18 min for intraoperative use and measured iPTH intraoperatively in 34 patients with renal hyperparathyroidism. Samples were taken before and 5 min and 15 min after parathyroid resection. Blood samples were put on ice immediately and sent to the hospital central laboratory via a pneumatic tube system. The first 76 probes were measured in parallel using three assays: the Nichols Quick PTH, the Roche Elecsys and the Biermann Immulite assay. The subsequent samples were only measured using the Elecsys assay. Determination of iPTH from 76 samples showed a correlation coefficient of 0.997 between the Immulite and Elecsys assay and a correlation coefficient of 0.987 for the Nichols Quick PTH and the Elecsys test. In renal hyperparathyroidism the mean iPTH was 26+/-2% of the starting value 5 min after subtotal parathyroidectomy and 18+/-2% after 15 min. Renal function influenced absolute iPTH values in patients with renal hyperparathyroidism but not relative changes. In patients with terminal renal insufficiency iPTH decreased from 615+/ 57 pg/m before preparation to 109+/-13 pg/ml 15 min after subtotal resection. In contrast in patients after kidney transplantation iPTH decreased from a lower starting value of 341+/-94 pg/ml to 58+/-9 pg/ml after 15 min. The iPTH kinetics showed a biphasic clearance of iPTH with an initial dominant half-life of 3.2 min and a terminal half-life of 29.2 min. Half-life did not correlate with renal function. All operations were successful as indicated by an adequate drop in PTH (from 709+/-92 pg/ml preoperatively to 22+/-6 pg/ml at discharge) and calcium (from 2.57+/-0.04 mmol/l to 2.32+/-0.04 mmol/l). In conclusion, intraoperative measurement of iPTH is also reliable in patients with renal hyperparathyroidism. Elimination kinetics are similar to that in patients with primary disease. However, the half-life was not influenced by renal function. The availability of a quick, inexpensive, routine iPTH test might expand its use to renal hyperparathyroidism, specifically for surgical decisions in problem cases. PMID- 11735019 TI - Paget's disease of the breast: a clinical perspective. AB - Paget's disease of the breast is a rare disorder of the nipple-areola complex often associated with an underlying in situ or invasive carcinoma. Eczematoid changes of the nipple-areola complex and persistent soreness or itching should raise suspicion of this disease. The histogenesis of Paget's disease of the breast continues to be debated and is important when considering treatment options. The epidermotropic theory suggests that Paget's cells are ductal carcinoma cells that have migrated from an underlying carcinoma of the breast parenchyma to the epidermis of the nipple. The in situ transformation theory has been proposed to explain the development of this disorder in patients in whom an underlying mammary carcinoma is not found or when there is an underlying carcinoma anatomically remote from the nipple-areola complex. Paget's cells are believed to arise as malignant cells in the epidermis of the nipple independent from any other pathologic process within the breast parenchyma. The current standard treatment of biopsy-proven Paget's disease involves mastectomy, although some studies have proposed the use of breast conservation therapy for patients in whom an underlying breast cancer cannot be located. We propose a treatment algorithm for patients presenting with Paget's disease of the breast. PMID- 11735021 TI - Insulator: from chromatin domain boundary to gene regulation. AB - An insulator or boundary element is a novel class of regulatory sequence and has properties consistent with a role in limiting enhancer activity; it helps to define the boundary between differentially regulated loci. Originally, an insulator was defined operationally by its ability to protect against positional effects and/or to block distal enhancer activity in transgenic assays, whereas recent experiments have focused on the role of insulators or boundaries in their native context. Many new features have been added to our concept of insulators, thereby revealing the complexity of their function. All these need to be accounted for by successful models. The research of insulator elements should provide new insights into the nature of enhancer function and gene regulation and provide an effective tool in applied gene therapy. PMID- 11735022 TI - Genomic structure of karyopherin alpha2 ( KPNA2) within a low-copy repeat on chromosome 17q23-q24 and mutation analysis in patients with Russell-Silver syndrome. AB - Human karyopherin alpha2 (KPNA2), a member of the karyopherin alpha family, plays a key role in the nuclear import of proteins with a classical nuclear localization signal (NLS). KPNA2, as part of a karyopherin alpha-beta heterodimer, directly binds to the NLS of proteins and functions as an adaptor that binds NLS-containing proteins via karyopherin beta to the nuclear pore complex. The NLS protein-receptor complex is translocated through the pore by an energy-dependent mechanism. Recently, we have identified and mapped the gene for KPNA2 in close proximity to a translocation breakpoint within 17q23-q24 associated with Russell-Silver syndrome (RSS). Therefore, we considered KPNA2 as a positional candidate gene for this heterogeneous disorder. RSS is mainly characterized by pre- and postnatal growth retardation, lateral asymmetry, and other dysmorphic features. Here, we present the genomic organization of the human KPNA2 gene with 11 exons spanning approximately 10 kb on chromosome 17q23-q24. Screening for mutations within all exons and adjacent intronic sequences from 31 unrelated RSS patients revealed three single nucleotide polymorphisms (SNPs) in exons 1, 5, and 7, and five SNPs in introns 1, 4 (2 SNPs), 8, and 9, respectively. No disease-related mutation was identified by comparing the sequence data of the RSS patients with their clinically normal parents and controls. PMID- 11735023 TI - Evaluation of denaturing high performance liquid chromatography (DHPLC) for the mutational analysis of the neurofibromatosis type 1 ( NF1) gene. AB - The identification of mutations in the NF1 gene causing type 1 neurofibromatosis (NF1) has presented a considerable challenge because of the large size of the gene, the lack of significant mutational clustering, the diversity of the underlying pathological lesions and the presence of NF1 pseudogenes. Denaturing high performance liquid chromatography (DHPLC), a high throughput, non-hazardous and largely automated heteroduplex-based technique, is in many ways ideally suited to mutation detection in this condition. DHPLC was therefore optimised for the rapid screening of the 60 exons and splice junctions of the NF1 gene in patients with NF1. The sensitivity of DHPLC was evaluated in a retrospective study of a cohort of 111 unrelated NF1 patients with known germline mutations; 97% of mutations were detected. In a subsequent prospective analysis of 50 unrelated NF1 patients, germline mutations were identified in 34 individuals (68%), 22 of these alterations being novel. This represents the highest rate of mutation detection so far reported for the NF1 gene with a single screening technique and genomic DNA as a target. PMID- 11735024 TI - A frameshift mutation in exon 28 of the OPA1 gene explains the high prevalence of dominant optic atrophy in the Danish population: evidence for a founder effect. AB - Dominant optic atrophy (DOA) is a hereditary optic neuropathy characterised by decreased visual acuity, colour vision deficits, centro-coecal scotoma and optic nerve pallor. The gene OPA1, encoding a dynamin-related GTPase, has recently been identified within the genetic linkage interval for the major locus for DOA on chromosome 3q28 and shown to harbour genetic aberrations segregating with disease in DOA families. The prevalence of the disorder in Denmark is reported to be the highest of any geographical location, suggestive of a founder effect. In order to establish the genetic basis of disease in a sample of 33 apparently unrelated Danish families, we screened DNA from affected members for OPA1 gene mutations by heteroduplex analysis and direct sequencing. A novel identical mutation in exon 28 (2826delT) was associated with DOA in 14 pedigrees and led to a frameshift and abnormal OPA1 protein -COOH terminus. Haplotype analysis of a region of approximately 1 Mb flanking the OPA1 gene using eight polymorphic markers revealed a common haplotype shared by all 14 patients; this haplotype was markedly over-represented compared with ethnically matched controls. Statistical analysis confirmed significant linkage disequilibrium with DOA over approximately 600 kb encompassing the disease mutation. We have therefore demonstrated that the relatively high frequency of DOA in Denmark is attributable to a founder mutation responsible for approximately 42% of the examined families and suggest that presymptomatic screening for the (2826delT) mutation may facilitate diagnosis and genetic counselling in a significant proportion of DOA patients of Danish ancestry. PMID- 11735025 TI - Mutational analysis of 85 mucopolysaccharidosis type I families: frequency of known mutations, identification of 17 novel mutations and in vitro expression of missense mutations. AB - The lysosomal storage disorder, mucopolysaccharidosis type I (MPS I), is caused by a deficiency of the enzyme alpha-L-iduronidase, which is involved in the breakdown of dermatan and heparan sulphates. There are three clinical phenotypes, ranging from the Hurler form characterised by skeletal abnormalities, hepatosplenomegaly and severe mental retardation, to the milder Scheie phenotype where there is aortic valve disease, corneal clouding, limited skeletal problems, but no mental retardation. In this study, 85 MPS I families (73 Hurler, 5 Hurler/Scheie, 7 Scheie) were screened for 9 known mutations (Q70X, A75T, 474 2a>g, L218P, A327P, W402X, P533R, R89Q, 678-7g>a). W402X was the most frequent mutation in our population (45.3%) and Q70X was the second most frequent (15.9%). In 30 families, either one or both of the mutations were not identified, which accounted for 25.9% of the total alleles. Therefore, all 14 exons of the alpha-L iduronidase gene were screened in these patients and 23 different sequence changes were found, 17 of which were previously unknown. The novel sequence changes include 4 deletions (153delC, 628del5, 740delC, 747delG), 5 nonsense mutations (Q60X, Y167X, Q400X, R619X, R628X), 6 missense mutations (C205Y, G208V, H240R, A319V, P496R, S633L), a splice site mutation (IVS12+5g>a), and a rare polymorphism (A591T). The polymorphism and novel missense mutations were transiently expressed in COS-7 cells and all of them except the polymorphism showed complete loss of enzyme activity. In total, 165 of the 170 mutant alleles were identified in this study and despite the high frequency of W402X and Q70X, the identification of many novel mutations unique to individual families further highlights the genetic heterogeneity of MPS I. PMID- 11735026 TI - Supravalvular aortic stenosis: genetic and molecular dissection of a complex mutation in the elastin gene. AB - We have identified two elastin gene (ELN) mutations located in cis in two related families with supravalvular aortic stenosis (SVAS). These mutations included an in-frame duplication in exon 18 (1034-1057dup) and a single base substitution in exon 26 (1829G-->A) predicted to result in the amino acid substitution R610Q. Haplotype analysis in one of the families identified an individual with a recombination between exon 18 and 26 of the elastin gene. This individual was unaffected and carried the exon 18 insertion mutation but not 1829G-->A. Skin fibroblasts were established from this recombinant normal individual and from an affected individual carrying both of the mutations. Reverse transcription/polymerase chain reaction (RT-PCR) analysis indicated that the expression of the mutant allele was reduced to 12%-27% of the normal allele in the affected but not in the unaffected individual. RNA-blot hybridization and immunoprecipitation experiments revealed reduced steady-state elastin mRNA levels and tropoelastin synthesis in the affected individual. RT-PCR analysis of the mRNA rescued by cycloheximide treatment indicated that mutation 1829G-->A created a cryptic donor splice site within exon 26, resulting in the deletion of four nucleotides at the 3'-end of exon 26 and a frameshift in the mRNA. This frameshift mutation generated a premature termination codon in the domain encoded by exon 28, clearly resulting in nonsense-mediated decay (NMD) of this frameshift RNA product. Despite considerable variability in the molecular nature of mutations responsible for SVAS, the unifying mechanism appears to be the generation of null alleles by NMD leading to elastin haploinsufficiency. PMID- 11735027 TI - Association of the mitochondrial DNA 5178 A/C polymorphism with serum lipid levels in the Japanese population. AB - As one approach to exploring whether the mitochondrial DNA 5178 adenine/cytosine (mt5178 A/C) polymorphism is associated with atherosclerosis, we genotyped 461 healthy Japanese individuals and studied the relationship of mt5178 A/C genotypes to serum lipid levels. Blood specimens were obtained after at least a 12-h fasting period from the subjects. The mt5178 A/C was genotyped by the polymerase chain reaction/restriction fragment length polymorphism method. The relative frequency of mt5178 A was 41.6% (192/461) and of mt5178 C was 58.4% (269/461). After adjustments for age and body mass index, the high-density lipoprotein cholesterol concentration in males carrying mt5178 A was significantly higher than that in males carrying mt5178 C ( P=0.026). The tryglyceride (TG) concentration in females carrying mt5178 A was significantly lower than that in females carrying mt5178 C ( P=0.012). This difference in the TG level between the two genotypes was more evident in postmenopausal females than in premenopausal females. Mt5178 A seems to have an antiatherogenic effect. This is the first genetic epidemiological report on the association of mt5178 A/C polymorphism with serum lipid levels in the Japanese population. PMID- 11735028 TI - Structure of human holocarboxylase synthetase gene and mutation spectrum of holocarboxylase synthetase deficiency. AB - Holocarboxylase synthetase (HLCS) is an enzyme that catalyzes the incorporation of biotin into apo-carboxylases, and its deficiency causes biotin-responsive multiple carboxylase deficiency. The reported sequences of cDNA for human HLCS from liver, lymphocyte, and KG-1 myeloid cell lines differ at their 5' regions. To elucidate variations of the human HLCS mRNA and longer 5' cDNA ends, we performed screening of the human liver cDNA library and rapid amplification of the cDNA ends (RACE). Our results suggest the existence of three types of HLCS mRNA that start at different exons. The first type starts at exon 1, and the second type starts at exon 3, and both are found in various human tissues. The third type, corresponding to the cDNA from the KG-1 cell, starts at exon 2 of the HLCS gene. Various splicing patterns from exons 3-6 were also observed. None of the variations of cDNA found created a new initiation codon. Mutation screening from exons 6-14, therefore, was sufficient to detect amino acid changes in HLCS in patients. Our direct sequencing strategy for screening mutations in the HLCS gene revealed mutations in five Japanese patients and seven non-Japanese patients. Our analyses involving 12 Japanese and 13 non-Japanese patients and studies by others indicate that (1) there is no panethnically prevalent mutation; (2) the Arg508Trp, Gly581Ser, and Val550Met mutations are found in both Japanese and non-Japanese populations; (3) the IVS10+5G-->A mutation is predominant and probably a founder mutation in European patients; (4) the 655-656insA, Leu237Pro, and 780delG mutations are unique in Japanese patients; (5) the spectrum of the mutations in the HLCS gene may vary substantially among different ethnic groups. PMID- 11735029 TI - Novel mutations of MYO15A associated with profound deafness in consanguineous families and moderately severe hearing loss in a patient with Smith-Magenis syndrome. AB - Mutations in myosin XVA are responsible for the shaker 2 ( sh2) phenotype in mice and nonsyndromic autosomal recessive profound hearing loss DFNB3 on chromosome 17p11.2. We have ascertained seven families with profound congenital hearing loss from Pakistan and India with evidence of linkage to DFNB3 at 17p11.2. We report three novel homozygous mutations in MYO15A segregating in three of these families. In addition, one hemizygous missense mutation of MYO15A was found in one of eight Smith-Magenis syndrome (del(17)p11.2) patients from North America who had moderately severe sensorineural hearing loss. PMID- 11735030 TI - Identification and characterization of two novel human mitochondrial elongation factor genes, hEFG2 and hEFG1, phylogenetically conserved through evolution. AB - Rapid progress in the sequencing of the genome of man and other species allows for the comparative analysis of their genetic structure and content. We have used a combined biochemical and computer-based approach to characterize a 146 kb human genomic bacterial artificial chromosome clone from chromosome 5q13 and discovered a novel human elongation-factor gene, hEFG2. The complete human EFG2 cDNA sequence is 3033 bp and contains 21 exons with conserved exon-intron splice junctions encompassing 45 kb of the genomic sequence with its 5'-end residing within a CpG island, characteristic of a housekeeping gene. The complete size of the hEFG2 cDNA was confirmed by Northern blot and reverse transcription/polymerase chain reaction analysis, which showed a single transcript of 3.2 kb ubiquitously expressed in various human tissues. The hEFG2 protein shows significant homology to several bacterial EF-G proteins, including that of Thermus thermophilus, and to the yeast Saccharomyces cerevisiae mitochondrial elongation factor-G ( MEF2). Multiple alignments reveal a novel gene family of mitochondrial EF-G proteins that can by divided into two subgroups, EF-G1 and EF-G2, in several eukaryotic species including S. pombe, Caenorhabditis elegans and Drosophila melanogaster. Using the information contained in the public databases, we also identified and cloned the complete coding sequence of the human EFG1 gene on chromosome 3q25. The cloning and characterization of these human mitochondrial elongation factor genes should permit us to address their role in the regulation of normal mitochondrial function and in various disease states. PMID- 11735031 TI - Analysis of short stature homeobox-containing gene ( SHOX) and auxological phenotype in dyschondrosteosis and isolated Madelung deformity. AB - Dyschondrosteosis (DCO; also called Leri-Weill syndrome) is a skeletal dysplasia characterised by disproportionate short stature because of mesomelic shortening of the limbs. Madelung deformity is a feature of DCO that is distinctive, variable in expressivity and frequently observed. Mutations of the SHOX (short stature homeobox-containing) gene have been previously described as causative in DCO. Isolated Madelung deformity (IMD) without the clinical characteristics of DCO has also been described in sporadic and a few familial cases but the genetic defect underlying IMD is unknown. In this study, we have examined 28 probands with DCO and seven probands with IMD for mutations in the SHOX gene by using polymorphic CA-repeat analysis, fluorescence in situ hybridisation (FISH), Southern blotting, direct sequencing and fibre-FISH analyses. This was combined with auxological examination of the probands and their family members. Evaluation of the auxological data showed a wide intra- and interfamilial phenotype variability in DCO. Out of 28 DCO probands, 22 (79%) were shown to have mutations in the SHOX gene. Sixteen unrelated DCO families had SHOX gene deletions. Four novel DCO-associated mutations were found in different families. In two additional DCO families, the previously described nonsense mutation (Arg195Stop) was detected. We conclude that mutations in the SHOX gene are the major factor in the pathogenesis of DCO. In a female proband with severe IMD and her unaffected sister, we detected an intrachromosomal duplication of the SHOX gene. PMID- 11735032 TI - A novel intronic mutation of the TAZ ( G4.5) gene in a patient with Barth syndrome: creation of a 5' splice donor site with variant GC consensus and elongation of the upstream exon. AB - Mutation analysis of the TAZ ( G4.5) gene was performed on a patient with Barth syndrome. The reverse transcription/polymerase chain reaction procedure showed aberrant splicing and elongation of exon 3 because of the insertion of 106 bases (IVS3+1 to +106) between exons 3 and 4. The genomic DNA revealed an intronic mutation four bases downstream from the new cleavage site (IVS3+110G-->A). The IVS3+110G-->A mutation created a novel 5' splice site that showed GC but not GT, and the additional splice site was used preferentially over the upstream authentic slice site. This is a new type of splicing mutation responsible for a human genetic disease. PMID- 11735033 TI - Genetic case-control association studies--correcting for multiple testing. PMID- 11735035 TI - Bax and other pro-apoptotic Bcl-2 family "killer-proteins" and their victim the mitochondrion. AB - Two major intracellular apoptosis signaling cascades have been characterized, the mitochondrial pathway and the death receptor pathway. The mitochondrial pathway is regulated by members of the Bcl-2 protein family. The members of this family can be subdivided into anti- and pro-apoptotic proteins. The pro-apoptotic members are further divided into two groups, the multidomain and the 'BH3 domain only' proteins. When cells are exposed to apoptotic stimulation, pro-apoptotic proteins are activated through post-translational modifications or changes in their conformation. The main site of action of the multidomain proteins are the mitochondria, where these proteins induce permeabilization of the outer membrane resulting in the release of proteins, including cytochrome c, from the intermembrane space. In the cytosol cytochrome c activates caspase cascades ultimately leading to cell death. Mounting evidence indicates that apoptosis is involved in a wide range of pathological conditions. Recent studies suggest that the mitochondrial signaling pathway is involved in several diseases. Although, so far, with the exception of C. elegans, most studies on apoptosis have been performed in mammalian systems, recently homologues to the Bcl-2 family members, including pro-apoptotic members, have been identified in Drosophila and zebrafish. Here the structure and function of the various pro-apoptotic Bcl-2 family members, their effects on mitochondria, and their involvement in diseases are discussed. PMID- 11735036 TI - Light- and electron-microscopic analysis of neuropeptide Y-immunoreactive amacrine cells in the guinea pig retina. AB - We investigated the morphology and synaptic connections of neuropeptide Y (NPY) containing neurons in the guinea pig retina by immunocytochemistry, using antisera against NPY. Specific NPY immunoreactivity was localized to a population of wide-field and regularly spaced amacrine cells with processes ramifying mainly in stratum 1 of the inner plexiform layer (IPL). Double-label immunohistochemistry demonstrated that all NPY-immunoreactive cells possessed glutamic acid decarboxylase 65 immunoreactivity. The synaptic connectivity of NPY immunoreactive amacrine cells was identified in the IPL by electron microscopy. The NPY-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in stratum 1 of the IPL. The most frequent postsynaptic targets of NPY-immunoreactive amacrine cells were other amacrine cell processes. Synaptic outputs to bipolar cells were also observed in a small number of cases. This finding suggests that NPY-containing amacrine cells may influence inner retinal circuitry in stratum 1 of the IPL, thus mediating visual processing. PMID- 11735037 TI - Changes in expression of P2 receptors in rat and mouse pancreas during development and ageing. AB - In view of the evidence for a role for extracellular ATP in both pancreatic endocrine and exocrine functions, we have investigated the expression of P2X and P2Y receptors in this tissue in neonate and aged rat and mouse. Using immunohistochemistry it was shown that P2X(1), P2X(4), P2X(7), P2Y(1) and P2Y(2) receptors were present in different regions of the rat and mouse pancreas; P2X(3) and P2X(6) receptors were not found, and P2X(5) immunolabelling was only found in some nerves. The pancreatic vasculature of both rat and mouse expressed P2X(1), P2X(2), P2Y(1) and P2Y(2) receptors in the smooth muscle. P2X(1) and P2X(4) receptors were absent in the islets of the neonate pancreas, but were progressively upregulated with age after birth. In contrast, the greatest expression of P2Y(1) in cells from the duct system was in neonate pancreas, while there was no P2Y(1) expression in aged rat pancreas. P2X(7) receptors had a consistent pattern of distribution in all of the groups examined, being located in the outer periphery of the islet. Using antibodies raised against insulin, somatostatin and glucagon, double-labelling immunofluorescence was used to identify P2X(7)-positive cells in different islet of Langerhans cell populations. Our results demonstrated a clear immunoreaction to P2X(7) receptors in islet alpha cells, while no P2X(7) was expressed in beta and delta cells. The significance of the differential expression of P2 receptors in the pancreas during development and ageing, and a possible role for the proliferation and death of the islet cell population are discussed. PMID- 11735038 TI - Sexually dimorphic neurogenesis is topographically matched with the anterior accessory olfactory bulb of the adult rat. AB - The accessory olfactory bulb (AOB) is a sexually dimorphic structure of the vomeronasal system, which plays a role in the control of sexual behaviors. In adult rats, we have demonstrated previously that the migrating neuroblasts of the subependymal layer (SEL) directed to the main olfactory bulb (MOB) also reach the AOB. To tackle the relation between sexual dimorphism and targeted cell migration, we quantified the neo-neurogenesis in the AOB of adult rats of both sexes. Our results confirm a morphological sexual dimorphism in the AOB granular layer volumes. We showed that the number of newly generated cells reaching the AOB in both sexes was considerable, even if lower than those directed to the MOB. Moreover, we demonstrated that the rate of neurogenesis in the anterior AOB of the two sexes was significantly different. PMID- 11735039 TI - Cannabinoid receptor CB1-like and glutamic acid decarboxylase-like immunoreactivities in the brain of Xenopus laevis. AB - Investigation of the cannabinoid system in a vertebrate group phylogenetically distant from mammals might improve understanding of its physiological role. Thus, in the present study, the distribution of the cannabinoid CB1 receptor has been investigated in the brain of Xenopus laevis (anuran amphibians) by immunohistochemistry, using both light and confocal laser-scanning microscopy. Immunostained neuronal perikarya and terminals were found in the olfactory bulb, dorsal and medial pallium, striatum, and amygdala. Varicosities and nerve terminals containing CB1-like immunoreactivity were also seen in the thalamus and hypothalamus. A number of stained cells were observed in the pars distalis of the pituitary gland. Positive nerve fibers were distributed throughout mesencephalic tegmentum, and in the cerebellum immunolabeling was observed in some Purkinje and possibly Golgi cells. The confocal microscopic analysis of CB1-like and glutamic acid decarboxylase-like immunoreactivities in both the medial pallium of the telencephalon and the olfactory bulbs showed a wide codistribution of the two markers. The present results indicate that distribution of CB1 is conserved in the course of phylogeny. Furthermore, the close relationship between CB1-like and glutamic acid decarboxylase-like immunolabelings point toward the existence of a functional link between cannabinergic and GABAergic innervations also in amphibian brain. PMID- 11735040 TI - Metastatic growth of squamous cell carcinomas is correlated with upregulation and redistribution of hemidesmosomal components. AB - Bullous pemphigoid antigen-1 (BPA1) and alpha(6)beta(4)-integrin colocalize at the hemidesmosomes in basal-layer keratinocytes of normal squamous epithelia. The expression of these genes was analyzed during the process of tumor cell invasion and metastasis on frozen sections of head and neck biopsies, and the structural appearance of hemidesmosomes was analyzed by electron microscopy. Despite a diminution of hemidesmosomal structures as revealed by electron microscopy, gene expression of BPA1 and alpha(6)beta(4)-integrins was distinctly upregulated with the onset of invasive growth, demonstrated at the mRNA level by in situ hybridization. The upregulated gene expression extended to the entire proliferative zone of invasive tumors, including the tumor cells which have lost contact with the basement membrane and no longer display hemidesmosomes. The polarized localization of the BPA1 and alpha(6)beta(4) proteins to the basal aspect of the peripheral tumor cells was largely retained in invasive but nonmetastatic lesions, but was lost upon progression to metastatic growth of head and neck squamous cell carcinomas (SCC), in which pericellular staining extended into many tumor cell layers. The results of this study confirm that expression of BPA1 and alpha(6)beta(4)-integrins is elevated in carcinoma cells but is not directed to intact hemidesmosomes. Importantly, this loss of directed localization is an indicator of the capacity to metastasize. PMID- 11735041 TI - Severity of oxidative stress generates different mechanisms of endothelial cell death. AB - The role of reactive oxygen species (ROS) in the pathogenesis of vascular diseases is well established, but few data exist on the mechanisms by which ROS induce endothelial cell (EC) death. We examined the conditions and the mechanisms by which oxidative stress induces EC death, using cultured confluent bovine aortic ECs exposed for 30 min to different concentrations of hydroxyl radicals (HO*) generated by hydrogen peroxide (H(2)O(2)) in the presence of 100 microM ferrous sulfate (FeSO(4)). Cell viability assays, Hoechst DNA staining, TUNEL (TDT-mediated dUTP-biotin nick end-labeling) analysis, agarose gel electrophoresis and annexin V assay were used to determine the effect of HO* on the viability of ECs, and to distinguish between apoptosis and necrosis. The results showed that at concentrations of up to 0.1 mM H(2)O(2)/FeSO(4), the large majority of cells are viable, except for approximately 12.5% death, which occurs by apoptosis. At a concentration of 0.2 mM H(2)O(2), the cell viability is reduced to 66%, while EC apoptosis remained at comparable values (14%). At high oxidative stress (0.5 mM H(2)O(2)), the cell viability was drastically reduced (approximately 39%), and the prevalent form of death was necrosis; apoptosis accounted for only approximately 17%. Together, these data indicate that: (1) HO* induce EC death either by apoptosis or necrosis and (2) the mechanisms of EC death differ as a function of the concentration of HO. Thus, the same insult can cause apoptosis and/or necrosis, as a function of the intensity rather than the nature of the insult. PMID- 11735042 TI - Immunocytochemical demonstration of the gap junction proteins connexin 43 and connexin 45 in the musculature of the rat small intestine. AB - The immunohistochemical localization of connexin (Cx) 43 and Cx 45 in the musculature of the rat small intestine was studied at the ultrastructural level, with special reference to the interstitial cells of Cajal in the deep muscular plexus region (ICC-DMP). Cx 43 was localized at gap junctions formed between every group of cells, i.e., smooth muscle cell~smooth muscle cell, smooth muscle cell--ICC-DMP and ICC-DMP--ICC-DMP. In contrast, Cx 45 immunoreactivity was only detected at gap junctions between ICC-DMP--ICC-DMP. Since different types of Cx molecules have different properties for electrical and chemical coupling of cells, it is suggested that the homotypic network of ICC-DMP connected with Cx 45 gap junctions may function as an independent compartment segregated from the whole cellular network including the smooth muscle cells connected with Cx 43 gap junctions. It is further speculated that the ICC-DMP of the rat small intestine communicate with each other and with smooth muscle cells via the passage of messenger molecules through Cx 43, but they may use an additional mechanism, as yet unknown, for communications restricted to other ICC-DMP. PMID- 11735043 TI - Localization of growth hormone receptors in rat and human thyroid cells. AB - Growth hormone (GH) exerts its multiple actions by binding to a specific receptor (GHR) widely distributed in the organism. It is well established that, in acromegaly, the thyroid gland is larger than normal and that GH increases triiodothyronin concentrations and decreases those of tetraiodothyronin (thyroxine). The aim of the present study was to analyze the presence of GHR and its mRNA in rat and human thyroid gland by Western blot, in situ hybridization techniques, and immunohistochemistry. A band of the expected size for GHR was shown in rat and human thyroid by Western blot. GHR immunoreactivity was found in virtually all follicles. The signal was mainly localized in the cytoplasm, although a nuclear positivity was also found. In situ hybridization techniques demonstrated the presence of GHR messenger RNA in the thyroid gland (cytoplasm of the follicular cells). These results provide direct morphological evidence that GHR is localized in the thyroid gland of mammals and opens up the possibility that GH regulates thyroid cell function directly or via local autocrine or paracrine production of insulin-like growth factor I. PMID- 11735045 TI - Prevention of neonatal estrogen imprinting by vitamin A as indicated by estrogen receptor expression in the mouse vagina. AB - Treatment of female mice with estrogen during the neonatal period induces estrogen-independent persistent proliferation and cornification of the vaginal epithelium when the animals become adults. However, the occurrence of such irreversible vaginal changes is blocked by concurrent retinol acetate (RA) treatment. This study aimed to determine the expression pattern of estrogen receptor (ER) alpha and beta in the vaginas of ovariectomized 35-day-old mice treated neonatally with 17beta-estradiol (E(2)) and/or RA. The amounts of ERalpha and ERbeta mRNA molecules in the vaginal RNA samples were determined by competitive reverse transcription/polymerase chain reaction. The levels of both mRNAs were lower in ovariectomized mice that had been treated neonatally with E(2) but not in those treated with E(2) plus RA. Neonatal E(2) treatment caused a decrease in the percentage of ERalpha-immunoreactive cells in the vaginal stroma during adulthood, and concurrent RA treatment inhibited the decrease. The amount of each ER mRNA was also measured in the vaginas of mature mice treated with E(2) and RA; no inhibitory activity of RA was seen in the mature mice. Our studies indicate that, in mouse vagina, the irreversible effects of neonatal imprinting by estrogen might be prevented by the simultaneous administration of vitamin A through the inhibition of a decrease of the number of ER-expressing cells. PMID- 11735044 TI - Maternal treatment with somatotropin during early gestation affects basic events of myogenesis in pigs. AB - The effects of maternal treatment with somatotropin during early gestation on fetal muscle development were determined. Crossbred gilts received daily injections of either 3 ml of a placebo ( n=31) or of 6 mg porcine somatotropin ( n=31) from day (d) 10 to 27 of gestation and samples were collected from d 28 embryos, d 37 and 62 fetuses, and from neonates. Administration of somatotropin increased the total number of fibres (primary and secondary fibres) in neonatal semitendinosus muscle of middle- and low-weight littermates, whilst no increase was observed in psoas major muscle. Somatotropin induced increases in muscular protein concentration, creatine kinase activity, muscle fibre girth, as well as type II to type I fibre conversion which revealed an advanced degree of differentiation at birth. Treatment effects on prenatal development preceded these changes. Increased DNA concentrations at d 28 of gestation indicate stimulation of cellular proliferation during the embryonic stages. Thereafter, the withdrawal of somatotropin caused a transient delay of differentiation as indicated by lower protein concentrations and creatine kinase activity compared with controls at d 37 of gestation. This was compensated again at d 62, and the number of semitendinosus primary fibres was increased in middle-weight fetuses, whereas secondary or total fibre number did not yet differ. However, enhanced expression of Myf5 and MyoD indicates higher numbers of initially determined, proliferating myoblasts that may have contributed to increased formation of secondary fibres. In conclusion, maternal somatotropin is an influential factor in early pregnancy capable of affecting the basic events of myogenesis. PMID- 11735047 TI - Pyrokinin/PBAN-like peptides in the central nervous system of Drosophila melanogaster. AB - The pyrokinin/pheromone biosynthesis activating neuropeptide (PBAN) family of peptides found in insects is characterized by a 5-amino-acid C-terminal sequence, FXPRLamide. The pentapeptide is the active core required for diverse physiological functions, including stimulation of pheromone biosynthesis in female moths, stimulation of muscle contraction, induction of embryonic diapause in Bombyx mori, and stimulation of melanization in some larval moths. Recently, this family of peptides has been implicated in accelerating the formation of the puparium in a dipteran. Using bioassay and immunocytochemical techniques, we demonstrate the presence of pyrokinin/PBAN-like peptides in the central nervous system of Drosophila melanogaster. Pheromonotropic activity was shown in the moths Helicoverpa zeaand Helicoverpa armigera by using dissected larval nervous systems and adult heads and bodies of D. melanogaster. Polyclonal antisera against the C-terminal ending of PBAN revealed the location of cell bodies and axons in the central nervous systems of larval and adult flies. Immunoreactive material was detected in at least three groups of neurons in the subesophageal ganglion of 3rd instar larvae, pupae, and adults. The ring gland of both larvae and adults contained immunoreactivity. Adult brain-subesophageal ganglion complex possessed additional neurons. The fused ventral ganglia of both larvae and adults contained three pairs of neurons that sent their axons to a neurohemal organ connected to the abdominal nervous system. These results indicate that the D. melanogasternervous system contains pyrokinin/PBAN-like peptides and that these peptides could be released into the hemolymph. PMID- 11735046 TI - Lipophorin of lower density is formed during immune responses in the lepidopteran insect Galleria mellonella. AB - Injection of heat-killed bacteria into larvae of the greater wax moth Galleria mellonella is followed by changes in lipoprotein composition in the hemolymph. Density gradient centrifugation experiments revealed that within the first four hours after injection, a part of larval lipoprotein, high-density lipophorin (HDLp), was converted into a lipoprotein of lower density. SDS-polyacrylamide gel electrophoresis analysis of the gradient fractions and sequencing of protein fragments, established that the exchangeable apolipoprotein apolipophorin III (apoLp-III), a potent immune-activator, was associated with this newly formed lipophorin. To investigate further the influence of lipophorin-associated apoLp III on immune-related reactions, we performed in vitro studies with isolated hemocytes from G. mellonella and lipophorins from the sphinx moth Manduca sexta, as a natural source of high amounts of low-density lipophorin (LDLp) and HDLp. The hemocytes were activated to form superoxide radicals upon incubation with LDLp, but not with HDLp. Fluorescence-labeled LDLp was specifically taken up by granular cells. This process was inhibited by adding an excess of unlabeled LDLp, but not by HDLp. We hypothesize that larval lipophorin formed in vivo is an endogenous signal for immune activation, specifically mediated by the binding of lipid-associated apoLp-III to hemocyte membrane receptors. PMID- 11735048 TI - Expression of carbamoylphosphate synthetase I and glutamine synthetase in hepatic organoids reconstructed by rat small hepatocytes and hepatic nonparenchymal cells. AB - The objective of this study was to investigate the expression of carbamoylphosphate synthetase I (CPS) and glutamine synthetase (GS) in small hepatocyte colonies and whether the heterogeneous expression of the enzymes could be induced during the maturation of small hepatocytes. Small hepatocytes isolated from an adult rat liver were cultured and proliferated to form colonies. The expression of CPS and GS was examined using immunocytochemistry and immunoblotting. In this culture more than 99% of morphologically hepatic cells were positive for CPS and all small hepatocytes were negative for GS at day 5. CPS-positive cells dramatically decreased with time in culture, whereas GS positive ones appeared and their number increased in the colonies. Two to 3 weeks after plating, colonies with rising and piled-up cells appeared and the number of such colonies reached about 25% of all colonies at day 30. In most rising and piled-up cells in colonies both proteins were strongly expressed, whereas many small hepatocytes in monolayer colonies did not express either protein. When small hepatocytes in monolayer colonies were overlayed with Matrigel, the cells gradually piled up and both CPS and GS proteins were dramatically induced. The expression of CPS and GS in small hepatocytes may interact with the extracellular matrix because the rising and piled-up cells appear to be induced by the extracellular matrix produced by hepatic nonparenchymal cells. PMID- 11735050 TI - The internal morality of clinical medicine: a paradigm for the ethics of the helping and healing professions. AB - The moral authority for professional ethics in medicine customarily rests in some source 'external' to medicine, i.e., a pre-existing philosophical system of ethics or some form of social construction, like consensus or dialogue. Rather, 'internal' morality is grounded in the phenomena of medicine, i.e., in the nature of the clinical encounter between physician and patient. From this, a philosophy of medicine is derived which gives moral force to the duties, virtues and obligations of physicians qua physicians. Similarly, an ethic specific to the other healing professions, law, teaching or ministry, can be derived from the specific ends to telos of each of these professions, which like medicine, are focused on a special type of human relationship. PMID- 11735051 TI - The internal morality of medicine: an evolutionary perspective. AB - A basic question of medical ethics is whether the norms governing medical practice should be understood as the application of principles and rules of 'the common morality' to medicine or whether some of these norms are 'internal' or 'proper' to medicine. In this article we describe and defend an evolutionary perspective on 'the internal morality of medicine' that is defined in terms of the goals of clinical medicine and a set of duties that constrain medical practice in pursuit of these goals. This perspective is developed by means of a critical examination of the 'essentialist' conception of the internal morality of medicine advocated by Edmund Pellegrino and the critique of internal morality approaches by Robert Veatch and Tom Beauchamp. PMID- 11735052 TI - Internal and external standards for medical morality. AB - What grounds and justifies conclusions in medical ethics? Is the source external or internal to medicine? Three influential types of answer have appeared in recent literature: an internal account, an external account, and a mixed internal/external account. The first defends an ethic derived from either the ends of medicine or professional practice standards. The second maintains that precepts in medical ethics rely upon and require justification by external standards such as those of public opinion, law, religious ethics, or philosophical ethics. The third claims that distinct medical ethics have emerged from distinct cultural frameworks, each with norms that govern physicians. There is merit in each perspective, but each over reaches its supporting arguments and fails to appreciate what is legitimate in the theses of its competitors. I propose a fourth account that offers a way to escape limitations of the other three, while retaining their most attractive features. PMID- 11735053 TI - The impossibility of a morality internal to medicine. AB - After distinguishing two different meanings of the notion of a 'morality internal to medicine' and considering a hypothetical case of a society that relied on its surgeons to 'eunuchize' priest/cantors to permit them to play an important religious/cultural role, this paper examines three reasons why morality cannot be derived from reflection on the ends of the practice of medicine: (1) there exist many medical roles and these have different ends or purposes, (2) even within any given medical role, there exists multiple, sometimes conflicting ends, and, most critically, (3) the ends of any practice such as medicine must come from outside the practice, that is, from the basic ends or purposes of human living. The paper concludes by considering whether these ends external to medicine are universally part of the moral reality or whether they are socially constructed. The paper argues that, even if various cultural accounts of the common, universal morality are 'socially constructed,' they may, nevertheless, be reflections, however, imperfect, of a more universal common morality that should be thought of as real. Therefore, the morality of medicine must come from a more fundamental morality external to medicine. That external morality will be socially constructed, but may nevertheless reflect an underlying common morality. PMID- 11735054 TI - A method in search of a purpose: the internal morality of medicine. AB - I begin this commentary with an expanded typology of theories that endorse an internal morality of medicine. I then subject these theories to a philosophical critique. I argue that the more robust claims for an internal morality fail to establish a stand-alone method for bioethics because they ignore crucial non medical values, violate norms of justice and fail to establish the normativity of medical values. I then argue that weaker versions of internalism avoid such problems, but at the cost of failing to provide a clear sense in which their moral norms are internal or can ground a comprehensive approach to moral problems. Finally, I explore various functions that an internal morality might serve, concluding with the observation that, while there may be a core of good sense to the notion of an internal morality of medicine, our expectations for it must be drastically lowered. PMID- 11735055 TI - Proper handling of soft tissue in the acute phase. AB - Proper handling of soft tissue in the acute phase demands proper attention to fundamental principles in wound evaluation, preparation, and repair. Foundational to immediate wound care is appropriate hemostasis and anesthesia. Correct soft tissue cleaning techniques are important, and the surgeon should understand special situations in which contamination may be a problem, including bites. The controversial issue of antibiotic prophylaxis is discussed. Certain wounds are not amenable to closure, but, where closure is indicated, orientation and preparation of the repair are important. Regional considerations important to the face are addressed. Numerous methods of wound closure are now available, and the advantages of each are enumerated. Postoperatively, attention to patient counseling, dressings, ointment, cleaning, and scar revision help assure an optimal outcome for the traumatized tissue. PMID- 11735056 TI - W-plasty and geometric broken line closure. AB - The decision to revise a scar should be based on risk versus benefit. Scars over 2 cm in length or scars greater than 2 mm in width can usually be improved with scar revision. Although many techniques exist, scar irregularization has been used for scar camouflage for many years. Two techniques commonly employed for scar irregularization are W-plasty and geometric broken line closure. W-plasty provides a regularly irregular scar, and geometric broken line closure provides an irregularly irregular scar. Each method can offer excellent results when performed correctly and in the appropriate situations. The advantages, disadvantages, and design of each method are discussed. PMID- 11735057 TI - Tissue expansion and serial excision in scar revision. AB - Scars in the head and neck caused by trauma or extirpative surgery can present a significant problem to the patient and may require revision. A variety of techniques, both surgical and nonsurgical, exists for treatment of unsightly scars. Surgical excision of scars relies upon recruitment of local tissue for closure of the ensuing defect. Tissue expansion and serial scar excision may be used to provide more tissue for advancement or local flap coverage of revised scars. Herein we review the history, technique, and complications of tissue expansion and serial excision as well as the basic scientific principles underlying these techniques. PMID- 11735058 TI - Scar revision via resurfacing. AB - Numerous techniques exist to treat noticeable facial scars. Techniques range from surgical excision to resurfacing. In this review of dermabrasion and laser resurfacing, we address the clinical considerations, techniques, adjuncts, and peri-operative management of scar resurfacing. Dermabrasion offers the advantage of being a tried-and-true technique familiar to surgeons. Recent advances in laser technology have resulted in the increased use of pulsed-dye lasers (PDLs), erbium:yttrium-aluminum-garnet (YAG) lasers, and CO(2) lasers. PDLs are effective for hypertrophic scars and show lower rates of recurrence compared with erbium:YAG and CO2 lasers. In contrast, erbium:YAG and CO(2) lasers are well suited to treating atrophic and acne scars. Chemical peels play a minor role in scar resurfacing and function primarily as an adjunct. Scar resurfacing is an integral part of scar camouflage and is often used in conjunction with excision and irregularization techniques. PMID- 11735059 TI - Scar contractures, hypertrophic scars, and keloids. AB - A scar contracture is the result of a contractile wound-healing process occurring in a scar that has already been reepithelialized and adequately healed. Keloids and hypertrophic scars (HTSs) are fibrous tissue outgrowths that result from a derailment in the normal wound-healing process. The exact incidence of keloids and HTSs remains unknown. Beyond the common belief that trauma is the initiating event of keloid and hypertrophic scar formation, the remainder of the process remains uncertain. A combination of biochemical factors, skin tension, endocrinologic factors, and genetic factors are the likely culprits. Treatment begins by educating the patient about the etiology of the scarring process. All treatment protocols are individualized, but the standard approach to keloids and HTSs begins with corticosteroid injection followed by surgical excision, pressure dressings, and long-term follow-up. PMID- 11735060 TI - Avoiding pitfalls and unfavorable outcomes in scar revision. AB - The goal of scar revision is to improve upon appearance and function. In addition, techniques used to rectify scars may recruit local "normal" tissue. Appropriate scar management begins at the time of injury. Initial treatment, patient selection, timing of repair, and technique selection are critical steps in the decision-making process. Weakness in any one step can lead to a worsened scar appearance. Some common difficult management issues in scar revision are outlined with the aid of select cases. PMID- 11735061 TI - Future management of scarring. AB - Future growth and development of plastic and reconstructive surgery will push forward on all fronts, from the prevention of injury to improvements in primary and secondary wound closure, healing, surgical planning, instrumentation, and techniques. Current technology is on the brink of promoting rapid healing and preventing scar formation at the cellular level by affecting the healing process. Tissue engineering has the potential of creating new tissue with the potential to closely approximate missing or damaged tissue from a biopsy of the original. The next generation of reconstructive surgeons may approach traumatic wounds in a completely different fashion, with computers, growth factors, and cell cultures as opposed to a scalpel and suture. PMID- 11735063 TI - Z-plasty: a concise review. AB - One of the most commonly used techniques in facial plastic surgery is the Z plasty. Main reasons to perform these transposition flaps are to lengthen a pre existing scar, to camouflage a scar, or to realign a scar. The classic 60 degrees Z-plasty allows a 75% increase in scar length and is the cornerstone against which all variations are compared. Understanding the classic Z-plasty permits the surgeon to expand his or her repertoire to include the numerous variations thereof. The double-opposing Z-plasty, unequal triangle Z-plasty, four-flap Z plasty, compound Z-plasty, and planimetric Z-plasty are the most frequent variants of the basic Z-plasty. Each are presented with illustrations and clinical indications. PMID- 11735064 TI - Local flaps in scar revision. AB - Local and regional skin flaps provide the foundation for reconstructive surgery of the face. When planned and executed properly, local skin flaps provide rapid reconstruction using donor tissue with excellent color and texture match, and a reliable, adequate blood supply. The majority of local skin flaps are utilized for reconstruction after superficial cancer extirpation, however, unfavorable scars from previous trauma, burns, or reconstruction efforts provide the motivated patient and surgeon another opportunity to restore form and function. This article reviews techniques commonly used to reconstruct the face and provides several cases where these techniques were successfully employed. PMID- 11735062 TI - Nonoperative techniques for scar management and revision. AB - Facial plastic surgeons are frequently called upon to manage facial scars, whether iatrogenic or traumatic. Numerous treatment modalities are available for scar management depending upon scar characteristics, age, and patient expectations. The focus of this article is to review commonly used nonsurgical methods of scar revision. These include topical applications (silicone, vitamin E, pressure dressing, herbal extracts), intralesional medication (steroids, antimitotics), soft tissue augmentation (collagen, fat), laser applications (585 nm flashlamp-pumped pulsed dye, CO(2)), cryotherapy, and make-up camouflage. Nonsurgical modalities can be used as prophylactic prevention of adverse scar formation, as definitive treatment, as intervening therapy until further surgical repair can be made, or as adjunctive treatment following surgical scar revision. PMID- 11735065 TI - [Prevention of diphtheria in Germany: yesterday, today, tomorrow - an overview]. AB - The year 2000 was the first without the registration of any cases of diphtheria in Germany. However, serologic protection against diphtheria amongst the population is very low, especially among young and middle-aged adults, where over 50 % are not or only minimally protected. In a review of the diphtheria cases of the last 7 years (n = 18), three factors were associated with the disease: adult age (14 cases between 18 and 50 years of age), travelling (in 14 of 18 cases travelling to or from endemic areas played a role) and occupational exposure (suspected in 6 of 18 cases). In Germany, the official guidelines which suggest a booster vaccination every ten years are followed very insufficiently. Should vaccination habits fail to improve, population immunity will further decrease and a resurgence of diphtheria will become more likely. Measures for future preventive activities are being discussed. PMID- 11735066 TI - [Health Status of German and Foreign Children: Why is Mehmet healthier than Maximilian?]. AB - At least in Germany social epidemiologists have practically never studied the association between the health status of children on one hand and their social status and nationality on the other. The census data from 1995 analysed here include data on social status (employment, social security, per capita income, school education) and nationality; and the 1 % sample of the population living in Germany covers all nationalities and all members of the families interviewed. The analysis includes 50,908 children aged 0-18 years and focuses on the health status and the smoking behaviour of the children and their parents. The result is somewhat surprising: In the age group 0-9 years foreign children seem to be more healthy than German children, despite the fact that their families more often belong to the lower status group and that their parents smoke more than the parents of the German children. It is hypothesised that social support in foreign families is stronger than in German families, and that this protective effect is stronger than the disadvantages of foreign children concerning social status of their families and smoking behaviour of their parents. PMID- 11735067 TI - [Immunisation rates of routine vaccinations for school starters as a quantitative measure in the vaccination behaviour]. AB - The aim of this retrospective long-term investigation was to compare the recommended routine vaccinations for school starters in the region of Dingolfing Landau, recording the results separately by birth years and comparing these with one another. It was to show to what extent changes in the quantitative measures in the vaccination behaviour were recorded over a period of one decade, respectively to what extent the current recommendations of any one time of the STIKO (Permanent Vaccination Commission of the Robert Koch-Institute) were reflected within each age group. For this purpose during the annual standardised examinations of school starters the rates of vaccinations were recorded according to the various birth years and sex, determined and anonomously evaluated. It was found that of the total eleven examined birth years the vaccinations in series of BCG was reduced continually from a commencing 50 % plus to under 10 %. Regarding vaccinations against diphtheria, tetanus and polio the frequency for the total examined period commenced at a rate of under 90 %, increasing to over 90 % for the birth years of 1989 and beyond. No child of the birth year of 1984 was completely vaccinated against HiB, whereas the rate of vaccinations rose relatively quickly so that by the birth year 1994 over 90 % of the school starters were completely immunised against this infectious disease. As far as the pertussis and MMR vaccinations were concerned the rate at the beginning of the observation period was between 40 % and 60 %, increasing continually in the following period to over 80 %. By the middle 'eighties hardly any school starters had been vaccinated against hepatitis B, whereas, in the last five examined birth years the rate increased relatively quickly to 64 % maximum. At the beginning of the observed period nearly every fifth school starter had, age related, complete vaccination protection, in respect of the then valid STIKO recommendations. At the end of this decade this applied only to every twelfth child. Due to the high rates of participation of school starters examinations carried out by the Public Child and Youth Health Service it is possible to determine the vaccination rates of children as it relates to the routine vaccinations carried out over a longer period and on a topical basis. As a result the vaccination behaviour, respectively the willingness of a relevant target group during that period can be shown quantitatively. As the investigations of the RKI (Robert Koch-Institute) have shown, the share of the strict opponents of vaccinations is merely between 1.5 % and 5 % resulting in the necessity for intensive educational work and the development of effective vaccination strategies which are needed in order to stabilise the rates of vaccinations, respectively to meet epidemiological requirements at a high level and to further improve the present rate. PMID- 11735068 TI - [Notification according to the law for protection against infections]. AB - Since January 1st 2001, the Law for Protection Against Infections decrees the duty of notification along two tracks, on the one hand by notification of infectious diseases and on the other hand by reporting the pathogens. However, the names of pathogens change due to scientific progress. Furthermore, these names include in some cases not only harmful agents of infectious diseases but also inoffensive organisms, i. e. Giardia lamblia. There are also problems in the notification of other pathogens such as Brucella sp., Campylobacter sp., Chlamydia psittaci,Clostridium botulinum, Cryptosporidium parvum, Leptospira interrogans, Listeria monocytogenes, Mycobacterium tuberculosis and similar agents, Norwalk-like viruses, Salmonella Paratyphi, Treponema pallidum, Trichinella spiralis and pathogenic serovars of Yersinia enterocolitica. Finally, the abandonment of notification for glanders, lymphogranuloma venereum, ulcus molle and variola appears to be risky, the reasons being partly unconvincing and contradictory. PMID- 11735069 TI - [How effective is the quality assurance as a result of external checking of nursing homes by the public health services]. AB - This is a report on the external checking of nursing homes by the Public Health Services in Duisburg, Germany, focussing on hygienic problems. For a second time in the last three-and-a-half years a total of 41 such homes were inspected and a total of 4150 residents were covered. Data on the intensity of care and on the hygienic standards were evaluated and compared with the results of 1998. 70.73 % of the homes inspected employed a nurse specifically trained in hygiene management (1998 15 %), 53.66 % used disinfectant textile cleaning and 34.39 % possessed written standards for management of infections relevant for nursing homes. 1.45 % of all residents had pressure sores III degrees and IV degrees classified according to Shea (1998: 3.37 %). In spite of significant improvements, deficits were still registered. To achieve further improvement attention should be focussed on the effectiveness of Public Health Services' activities. Increased co-operation with other institutions engaged in the external checkings as well as a closer focus on specific subjects in hygiene could be appropriate. PMID- 11735070 TI - [Surveillance of gastroenteritic infections in lower Saxony: results and experiences from a regional public-health project over six years]. AB - In Lower Saxony, a regional public health project on a pathogen-specific surveillance of Enteritis infectiosa was carried out from 1994 to 2000. In a separate reporting procedure thirteen health departments transmitted data on the specific pathogen collected from laboratory reports for a joint analysis. The results were distributed among the participants, providing them with information on the situation and trends in their districts and in addition giving them the possibility to compare their data with those of other districts. The incidence of Enteritis infectiosa was 199 reported cases per 100,000 person years. Salmonella was reported most frequently (51 %; S. enteritidis: 33 %, S. TYPHIMURIUM: 14 %, 111 other serotypes: 4 %), followed by Campylobacter (18 %), Rotavirus (17 %), Yersinia enterocolitica (6 %) and Adenovirus (5 %). The incidences of Salmonella and Yersinia enterocolitica were found to be nearly constant over the project period while the incidences of Campylobacter, Rotavirus and Adenovirus increased. This, however, may be due to intensified diagnostics and better reporting behaviour rather than to a real increase. The incidence rates and their trends differed from other German states. This observation emphasizes the need for a regional analysis of infectious disease surveillance data. PMID- 11735071 TI - [Remarks to statement from the rate of cause of death due to ischaemic heart diseases]. AB - The rate of cause of death due to IHD ('ischaemic heart diseases') represents a terminal occurrence of the arteriosclerotic basic disease. lts extent is determined by the incidence of cardiovascular diseases, prevalence and duration of sickness. The relative frequency of falling ill as well as the reflection of the risk of falling ill and the extent of the population fallen ill are not known at all or only very insufficiently. Thus the question cannot be answered how the risk to fall ill with the coronary heart disease must be considered in view of the high mortality rate IHD of the population in the total mortality of the population. There are recognizable influences on the rate of cause of death due to IHD, such as the duration of sickness, the increasing average life expectancy and the procedure of coding death certificates and the selection of the basic disease. So the abrupt increase of the infarct mortality after the turn in the new federal states, as well as in Mecklenburg-Western Pomerania is to be attributed to the adoption to the coding procedure. The increasing trend of the mortality rate of IHD up to 1995 in Mecklenburg-Western Pomeranian infers an individual shortening of the sickness of existing coronary heart diseases among the population. After this temporary increase of infarct mortality the decrease of this figure suggests prolongation of the duration of sickness. The rate of cause of death from 'ischaemic heart diseases' also represents a terminal occurrence in life. The development of specific mortality in advanced age leads to the observation that the emphasis within the cause of death structure among those classifications of the elderly at an extremely advanced age, gradually shifts towards mortality caused by ischaemic heart disease. From the accompanying decreasing trend of mortality from acute coronary events in the younger and medium age classes a decrease in the total mortality can be concluded. Even the search for diagnosis during post mortems is reflected in the cause of death statistics, which can be evidenced on the basis of comparisons made between the figures of death from 'ischaemic heart diseases' and from 'senility, other symptoms, and imprecisely described conditions'. The difference between the original federal states and the new federal states of Eastern Germany, as also in the Federal State of Mecklenburg-Western Pomerania is obvious. Summing up, it can therefore be said, that the cause of death rate from 'ischaemic heart diseases' is only of limited significance as regards the hazards from the underlying arteriosclerotic disease of the blood vessels. lt can only be assessed in connection with the age structure of the population and the structure of all causes of death. The validity of the cause of death from 'ischaemic heart diseases' is therefore not very great in reporting practice. PMID- 11735072 TI - [The influence of regional differences in drinking style on hazardous use, excessive drinking, abuse and dependence]. AB - INTRODUCTION: Previous research in Germany identified a "north-south" gradient in the hazardous use of alcoholic drinks, but this had methodological limitations. The present study estimates the hazardous use of alcohol more reliably. In addition, regional differences in drinking style and their relation to indicators of problematic alcohol use are examined. METHODS: Data are from 1995 and 1997 representative samples of the non-institutionalised German general population aged 18 to 59 years. Based on 15,017 complete observations prevalence rates for abstinence, hazardous consumption (>30 g ethanol per day for men; >20 g ethanol per day for women), two or more CAGE-symptoms, and heavy episodic drinking (5+ drinks consumed in a day) were calculated and a cluster analysis was performed with regard to the 16 German federal states. RESULTS: Regional differences exist in the prevalence of abstinence and hazardous drinking, but a "north-south" gradient was not evident. However, cluster analysis results suggest a "north south" difference in drinking style, i.e. the quantity-frequency pattern of beer, wine, and spirits consumption. Compared with the northern German federal states, those in the south experienced more beer and less wine and spirits consumption in combination with a lower frequency of drinking beer and spirits. No cluster difference was found for hazardous drinking, lifetime prevalence of 2+ CAGE items or consumption of 5+ drinks in a day. Yet, mean consumption of ethanol was significantly higher in the north. CONCLUSIONS: Research suggesting more hazardous consumption in southern Germany must be tempered by these findings that found no 'north-south' gradient in several different indicators of problematic alcohol use. Different drinking styles in northern and southern states do not result in differences concerning prevalence of binge drinking nor alcohol-related problems. Drinking style only made a difference in the mean consumption of ethanol. Methodological differences between the reported studies are discussed. PMID- 11735074 TI - Around my table. PMID- 11735075 TI - Being met--a passageway to hope for relatives of patients at risk of committing suicide: a phenomenological hermeneutic study. AB - Fifteen Norwegian relatives narrated their experiences of "being met" by mental health care personnel regarding the care of their adult family member who had seriously thought about or attempted suicide. The narrative interviews were audiotaped, transcribed into text, and then interpreted using a phenomenological hermeneutic method. The results of the interpretation revealed that the context of being met was characterized by "being helpless and powerless." The six themes that were constructed describing being met were: Being-seen as a human being; participating in an I-Thou relationship; trusting personnel, treatment, and care; feeling trusted by personnel; being consoled; and entering into hope. The experience of being met in the midst of the care of a relative in crisis created possibilities for hope. Being met, in this context, is a passageway to hope. PMID- 11735076 TI - The effects of group intervention for battered women in Korea. AB - This study developed a group intervention model appropriate for battered women in Korea and tested its effectiveness. The sessions in the group intervention were formatted to stress the following topics: assess trauma, identify major problems, deal with feelings, understand self, identify batterer's characteristics, improve stress management strategies, develop action plans, and promote empowerment. The major finding was that the trait anxiety scores of the 16 battered women in the experimental group significantly decreased after the intervention. The change in levels of state anxiety, self-esteem, and depression in the experimental group were not significantly different from those of the 17 the subjects in the control group. PMID- 11735077 TI - Assessment of inpatient treatment of patients with schizophrenia in Japan: implications for practice. AB - This descriptive study examined a stratified random sample by month of 202 persons with schizophrenia hospitalized on an inpatient unit at a nonprofit hospital located in Tokyo, Japan in 1998. This is one of the first studies reported that evaluates the inpatient care provided to persons with schizophrenia. The average length of stay was 107 days. The master treatment largely focused on medication management with other domains including daily observations and documentation of care not meeting the standard of care. Over 25% of the patients were in restraints during their hospitalization. Two patient deaths were reported. Problematic areas were patient teaching, monitoring of medication side effects, treatment planning, and discharge planning. The study site is fully accredited and attending physicians were all psychiatrists. How pervasive these deficiencies are in other settings is not known. The findings of the study were shared with the professional staff. PMID- 11735078 TI - A comparison of effectiveness and cost between two models of care for individuals with schizophrenia living in Taiwan. AB - Schizophrenia is a chronic disease that places tremendous costs and burden on patients, families, and communities. The purpose of this study was to compare the cost and effectiveness of a hospital-based home care model and a traditional community care model for schizophrenia in Taiwan using six principles of cost effectiveness analysis. Based on the health care provider's analytic perspective, four effectiveness indicators and four service costs were identified and measured, and the ratio of average cost value to effectiveness score for each patient was calculated. According to that ratio, the hospital-based home care model was more cost-effective. The results suggest that the hospital-based outreach home care model is a cost-effective way to care for patients and their family in the community. PMID- 11735079 TI - Predictors of pain catastrophizing in women with rheumatoid arthritis. AB - Catastrophizing has been conceptualized as an appraisal, a cognitive distortion, and a coping mechanism in the psychosocial literature. Regardless of its conceptualization, catastrophizing has been associated with negative psychological and physical outcomes in numerous studies, including our intervention study with 90 women with rheumatoid arthritis. Because of catastrophizing's robust relationship with negative outcomes, predictors of catastrophizing as a pain coping behavior were investigated in this sample, using data collected from two points in time before the intervention. Using Lazarus and Folkman's stress and coping theoretical framework to guide the analyses, variables with a proximal relationship to catastrophizing in the framework were examined for significant associations with pain catastrophizing using correlational analyses. Subsequent stepwise regression involving all variables from Time 1 with significant associations resulted in a model that explained 63% of the variance in Time 2 pain catastrophizing scores. The four predictors in this model were dispositional pessimism, passive pain coping, venting (as a pain coping behavior), and arthritis helplessness. Potential clinical implications related to these predictors are also discussed. Knowledge about predictors of catastrophizing may enhance efforts to address this maladaptive pattern through educational and therapeutic approaches. PMID- 11735080 TI - Are patients with Hirschsprung's disease prediabetic? AB - Hirschsprung's disease (HD) is considered a focal disease usually confined to the distal colon and rectum. However, autonomic dysfunction and dysmotility in the upper gastrointestinal tract have been reported, suggesting that this disease is not only confined to the distal gastrointestinal tract. This study examines the fasting and postprandial levels of glucose and insulin in adult patients with HD to elucidate whether there might also be an endocrine involvement in this disease. Sixteen patients with surgically treated HD during early childhood and 17 healthy subjects were studied. All subjects ingested a caloric liquid meal containing glucose, lactose, maize oil, and water (2,020 kJ) after an overnight fast. Blood samples were collected at regular intervals for insulin and glucose analyses. Fasting levels of both glucose (P <.05) and insulin (P <.02) were significantly higher in patients compared with healthy controls. Peak concentration of insulin following meal intake was significantly higher in the patient group (P <.05), and peak concentration of glucose tended to be higher in patients compared with controls (P =.06). There was no correlation between body mass index and serum levels of glucose or insulin. The present study shows that adult patients treated for HD during childhood have an impaired glucose and insulin homeostasis, indicating a mild degree of insulin resistance. This may imply susceptibility towards development of non-insulin-dependent diabetes mellitus. PMID- 11735081 TI - Obesity gene variant and elite endurance performance. AB - beta2-adrenergic receptor (ADRB2) Gln27Glu genotype was determined in sedentary (n = 19), active (n = 20), and elite endurance athletic (n = 24) Caucasian postmenopausal women. Age was similar in all physical activity and ADRB2 genotype groups. ADRB2 genotypes were in Hardy-Weinberg equilibrium in sedentary and active women, but not in the athletes (chi(2) = 4.28, P <.05), due to the near absence of ADRB2 Glu27Glu homozygotes among the athletes. Weight tended to be higher in ADRB2 Glu27Glu women (63.5 +/- 1.8 v 57.7 +/- 1.7 and 60.0 +/- 1.8, P =.08), as did body mass index (BMI) (25.0 +/- 0.4 v 22.9 +/- 0.6 and 23.4 +/- 0.5 kg/m(2), P =.05), due to a higher fat mass in Glu27Glu women (24.1 +/- 1.0 v 18.1 +/- 1.4 and 20.1 +/- 1.4 kg, P <.05). Maximal O2 consumption was lower in ADRB2 Glu27Glu than in ADRB2 Glu27Gln and Gln27Gln genotype women (25.4 +/- 1.1 v 32.4 +/- 1.5 and 29.1 +/- 1.7 mL/kg/min, P <.05). We conclude that the Glu27Glu ADRB2 genotype may dissociate from and the Gln27Gln and Gln27Glu genotypes may associate with elite endurance performance in older women. PMID- 11735082 TI - Platelet Na,K-adenosine triphosphatase as a tissue marker of hyperthyroidism. AB - Platelet Na(+),K(+)-adenosine triphosphatase (ATPase) activity was measured in 34 (15 males, 19 females) healthy subjects, 89 (35 males, 54 females) hyperthyroid patients, and 34 (7 males, 27 females) treated hyperthyroid patients to assess the potential of this measurement as a tissue marker and diagnostic test for hyperthyroidism. Platelet Na(+),K(+)-ATPase activity was measured in platelet lysates by the rate of release of phosphate from adenosine triphosphate (ATP) in the presence and absence of ouabain. Platelet Na(+),K(+)-ATPase activity (median and range) in the hyperthyroid group (271, 169 to 821 pmol/h/g protein) was significantly higher compared with the healthy group (125, 74 to 185 micromol/h/g protein, P <.001 by Mann-Whitney U test). The treated hyperthyroid group had slightly, but significantly higher, free triiodothyronine (FT3) and free thyroxine (FT4), as well as platelet Na(+),K(+)-ATPase activity (147, 98 to 246 micromol/h/g protein, P <.05). If a platelet Na(+),K(+)-ATPase activity of 190 micromol/h/g protein was used as a cut off value, the specificity and sensitivity were 90% and 93%, respectively. We conclude that platelet Na(+),K(+)-ATPase may be a useful tissue marker of hyperthyroidism. PMID- 11735083 TI - Effects of severity of chronic obstructive pulmonary disease on thyroid function. AB - To investigate thyroid function in chronic obstructive pulmonary disease (COPD), 46 consecutive patients (35 men) with stable, mild-to-severe disease, having a mean (SD) age of 67 +/- 7 years were studied. All subjects underwent pulmonary function tests (PFTs), arterial blood gas determination, and measurement of serum total thyroxine (TT4), total triiodothyronine (TT3), resin T3 uptake (RT3U), reverse triiodothyronine (rT3), and thyroid-stimulating hormone (TSH) levels. The free thyroxine and free triiodothyronine indexes (FT4I = RT3U/30TT4 and FT3I = RT3U/30TT3, respectively) along with the TT3/TT4 ratio were calculated; the latter was used as a marker of peripheral conversion of thyroxine into triiodothyronine. Interleukin (IL)-6 was also measured to evaluate its potential associations with thyroidal hormone levels. On the basis of forced expiratory volume in 1 second (FEV1), patients were divided in 2 groups: group 1, (FEV1 > or = 50% of predicted, n = 26), with mild-to-moderate COPD and group 2 (FEV1 < 50% of predicted, n = 20) having severe disease. All subjects had normal serum thyroid hormone levels; for the entire COPD population, mean values were 7.80 +/- 1.60 microg/dL for TT4, 1.12 +/- 0.20 ng/mL for TT3, 29.0 +/- 1.88 for RT3U, 7.54 +/- 1.34 for FT4I, 1.07 +/- 0.16 for FT3I, 18.71 +/- 5.89 ng/dL for rT3, and 1.15 +/- 0.6 microU/mL for TSH. Mean TT3/TT4 ratio was 0.14 +/- 0.03. In group 1, TT3, TT4, and TT3/TT4 ratio did not correlate with age, FEV1, PaO2, or inhaled corticosteroids. Similarly, in group 2, TT3 and TT4 were unrelated to the above mentioned variables; however, there was a strong positive correlation between TT3/TT4 ratio and PaO2 (r =.61, P =.004). IL-6 was within normal limits in all subjects, and it did not correlate with any thyroid hormone either in group 1 or in group 2. It is concluded that in stable COPD, severity of disease through hypoxemia is important in determining the peripheral metabolism of thyroid hormones. Whether this constitutes an adaptation is not known. PMID- 11735084 TI - Delta opioid receptors mediate glucose uptake in skeletal muscles of lean and obese-diabetic (ob/ob) mice. AB - Specific binding sites for [125I]beta-endorphin and the delta1-opioid [3H][D pen(2), D-pen(5)]enkephalin (DPDPE) were quantified using autoradiography in soleus and extensor digitorum longus (EDL) muscles of lean and obese-diabetic (ob/ob) mice. The density of binding was significantly higher in obese-diabetic than lean mice. The uptake of 2-deoxy-D-[1-3H]deoxyglucose, a nonmetabolized glucose analogue, into isolated soleus and EDL muscles was stimulated by beta endorphin, beta-endorphin 1-27, and DPDPE, but not by the delta2-opioid deltorphin II. Both beta-endorphin and DPDPE stimulated deoxyglucose uptake in obese-diabetic mice. Thus, glucose transport in skeletal muscle may be partly mediated via delta1-opioid receptors. The increased receptor density in obese diabetic mice may be an adaptive response. PMID- 11735085 TI - Magnesium reduces insulin-stimulated glucose uptake and serum lipid concentrations in type 1 diabetes. AB - A magnesium (Mg) deficit has been described in patients with type 1 diabetes, and it has been related to the development of cardiovascular disease. We tested the hypothesis that type 1 diabetic patients have deficits in dietary Mg intake and that proper long-term (24 weeks) oral Mg supplementation would reduce cardiovascular risk factors. Therefore, the Mg status, dietary Mg intake, and the effect of Mg supplementation were evaluated in 10 type 1 diabetic patients and 5 control subjects. Muscle Mg content was decreased by 7% in the type 1 diabetic patients, and it increased by 5% after 24 weeks of oral MgO supplementation. Acute and chronic Mg supplementation decreased serum total cholesterol, serum low density lipoprotein (LDL)-cholesterol, and apolipoprotein B. Insulin-stimulated glucose uptake decreased by 35% after 24 weeks of oral MgO supplementation. Eight of 10 patients with type 1 diabetes had a daily intake of Mg below 90% of the recommended daily allowance. In conclusion, a Mg deficit was found in type 1 diabetic patients. The deficit might be due partly to a relatively Mg-deficient diet. Mg repletion was associated with a decrease in atherogenic lipid fractions and a reduced insulin-stimulated glucose uptake. PMID- 11735086 TI - Long-term effect of partial nephrectomy on biological parameters, kidney histology, and guanidino compound levels in mice. AB - The long-term adverse consequences of early renal mass reduction in mice have not yet been investigated. The effects of partial surgical nephrectomy (NX) in 2 month-old mice on some biological parameters, on histopathologic and morphometric features of the kidney, and on urea and guanidino compound (GC) levels in plasma, urine, and brain were examined at 10 days, and 1, 2, 4, and 12 months postsurgery. Body weight, urinary volume, and plasma urea were most affected at 10 days and 12 months post-NX. NX-induced changes in the remaining renal tissue (including hypertrophy, glomerular mesangial expansion, and presence of protein casts) increased with age. As in human renal insufficiency, NX mice showed significantly higher plasma guanidinosuccinic acid (GSA) and creatinine (CTN) levels at all studied periods. The same tendency could be seen for most other plasma GCs examined, except for arginine (Arg), guanidinoacetic acid (GAA), and homoarginine (HA). As seen in human pathobiochemistry, the latter 2 compounds tended to be lower in NX mice in our follow-up study. Remarkably, and also similar to humans, NX mice excreted less GAA and more GSA than controls during the entire follow-up study. During the follow-up, excretion levels of GAA were unchanged in NX and sham-operated mice. In brain, GAA and gamma-guanidinobutyric acid (GBA) levels were always higher in NX mice with a tendency to respectively increase or decrease over time in NX as well as sham-operated mice. Although urea and GC metabolism were influenced by time post-NX and aging, the model was confirmed to display a mild stable chronic impairment of renal function. Histopathologic and morphometric changes of the kidney increased with age. PMID- 11735087 TI - Urinary excretion of olive oil phenols and their metabolites in humans. AB - We have recently demonstrated, in humans, the bioavailability of hydroxytyrosol (3,4-dihydroxyphenylethanol; HT), one of the major antioxidant components of virgin olive oil. In particular, we reported that this compound is present in lipoproteins involved in atherosclerotic processes and is excreted in the urine mainly as glucuronide-conjugate. The aim of the present study was to elucidate, in humans, the metabolic fate of HT after ingestion of virgin olive oil. After administration of virgin olive oil, 24-hour urine collections of healthy volunteers were prepared for gas chromatography-mass spectrometry analyses in order to identify and quantify HT and its metabolites homovanillic alcohol (HVA1c) and homovanillic acid (HVA). The results indicate that this compound undergoes the action of catechol-O-methyl transferase (COMT), enzymes involved in the catecholamine catabolism, resulting in an enhanced excretion of HVA1c. We also found a significant increase of HVA, indicating an oxidation of the ethanolic residue of HT and/or of HVA1c in humans. The excretion of both metabolites significantly correlated with the dose of administered HT. PMID- 11735088 TI - Comparison of creatine ingestion and resistance training on energy expenditure and limb blood flow. AB - This study determined the effects of 28 days of oral creatine ingestion (days 1 to 5 = 20g/d; [5 g 4 times daily]: days 6 to 28 = 10 g/d; [5 g twice daily]) alone and with resistance training (5 hours/week) on resting metabolic rate (RMR), body composition, muscular strength (1RM), and limb blood flow (LBF). Using a double-blind, placebo-controlled design, 30 healthy male volunteers (21 +/- 3 years; 18 to 30 years) were randomly assigned to 1 of 3 groups; pure creatine monohydrate alone (Cr; n = 10), creatine plus resistance training (Cr RT; n = 10), or placebo plus resistance training (P-RT; n = 10). Body composition (DEXA, Lunar DPX-IQ), body mass, bench, and leg press 1RM (isotonic), RMR (indirect calorimetry; ventilated hood), and forearm and calf LBF (venous occlusive plethysmography) were obtained on all 30 subjects on 3 occasions beginning at approximately 6:00 AM following an overnight fast and 24 hours removed from the last training session; baseline (day 0), and 7 days and 29 days following the interventions. No differences existed among groups at baseline for any of the variables measured. Following the 28-day interventions, body mass (Cr, 73.9 +/- 11.5 v 75.6 +/- 12.5 kg; Cr-RT, 78.8 +/- 6.7 v 80.8 +/- 6.8 kg; P <.01) and total body water (Cr, 40.4 +/- 6.8 v 42.6 +/- 7.2 L, 5.5%; Cr-RT, 40.6 +/- 2.4 v 42.3 +/- 2.2 L, 4.3%; P <.01) increased significantly in Cr and Cr-RT, but remained unchanged in P-RT, whereas, fat-free mass (FFM) increased significantly in Cr-RT (63 +/- 2.8 v 64.7 +/- 3.6 kg; P <.01) and showed a tendency to increase in Cr (58.1 +/- 8.1 v 59 +/- 8.8 kg; P =.07). Following the 28-day period, all groups significantly increased (P <.01) bench (Cr, 77.3 +/- 4 v 83.2 +/- 3.6 kg; Cr-RT, 76.8 +/- 4.5 v 90.5 +/- 4.5 kg; P-RT, 76.0 +/- 3.4 v 85.5 +/- 3.2 kg), and leg press (Cr, 205.5 +/- 14.5 v 238.6 +/- 13.2 kg; Cr-RT, 167.7 +/- 13.2 v 238.6 +/- 17.3 kg; P-RT, 200.5 +/- 9.5 v 255 +/- 13.2 kg) 1RM muscular strength. However, Cr-RT improved significantly more (P <.05) on the leg press 1RM than Cr and P-RT and the bench press 1RM than Cr (P <.01). Calf (30%) and forearm (38%) LBF increased significantly (P <.05) in the Cr-RT, but remained unchanged in the Cr and P-RT groups following the supplementation period. RMR expressed on an absolute basis was increased in the Cr (1,860.1 +/- 164.9 v 1,907 +/- 173.4 kcal/d, 2.5%; P <.05) and Cr-RT (1,971.4 +/- 171.8 v 2,085.7 +/- 183.6 kcal/d, 5%; P <.05), but remained unchanged from baseline in P-RT. Total cholesterol decreased significantly in Cr-RT (-9.9%; 172 +/- 27 v 155 +/- 26 mg/dL; P <.01) compared with Cr (174 +/- 46 v 178 +/- 43 mg/dL) and P-RT (162 +/- 32 v 161 +/- 36 mg/dL) following the 28-day intervention. These findings suggest that the addition of creatine supplementation to resistance training significantly increases total and fat-free body mass, muscular strength, peripheral blood flow, and resting energy expenditure and improves blood cholesterol. PMID- 11735089 TI - Increased urinary type IV collagen marks the development of glomerular pathology in diabetic d/db mice. AB - The diabetic db/db mouse exhibits increased albumin excretion soon after the onset of obesity and hyperglycemia, and later manifests glomerular mesangial matrix expansion resembling that found in human diabetic nephropathy. Since the glomerular lesion in this rodent model of type 2 diabetes is associated with renal overexpression of mRNA encoding type IV collagen, we postulated that changes in the urinary excretion of collagen IV may reflect developing glomerular pathology. To explore this hypothesis, we monitored urinary collagen IV (measured by immunoassay) in db/db mice during the course of evolution of nephropathy. At age 8 weeks, collagen IV excretion was not different in diabetic compared to nondiabetic animals despite marked albuminuria, but was significantly increased in db/db compared to db/m mice at age 12 and 16 weeks. Serum levels of collagen IV did not significantly differ between normal versus diabetic mice at any age. Glomerular morphometry revealed mesangial matrix expansion at age 12 weeks, coincident with the rise in collagen IV excretion, which became more marked at age 16 weeks in association with reduced creatinine clearance and elevated serum creatinine. The findings suggest that increased urinary type IV collagen is a better indicator than albuminuria of developing glomerular matrix accumulation that results in compromised renal filtration function. PMID- 11735090 TI - Trends of lipoprotein variables from childhood to adulthood in offspring of parents with coronary heart disease: the Bogalusa Heart Study. AB - Although dyslipidemia among offspring of parents with coronary heart disease (CHD) has been known, the development of this adverse relationship with respect to specific lipoprotein variables from childhood to young adulthood has not been elucidated. This aspect was examined in a young adult cohort with (n = 271) and without (n = 805) a parental history of CHD followed longitudinally since childhood by repeated surveys from 1973 to 1991. Trends in fasting lipoprotein variables by parental CHD status were assessed by Lowess smoothing curve and Generalized Estimating Equations (GEE). In multivariate analyses adjusted for race and sex, parental CHD associated positively with low-density lipoprotein cholesterol (LDL-C, P <.01) and triglycerides (P <.05) mainly at the young adulthood age, whereas a positive association was noted with very-low-density lipoprotein cholesterol (VLDL-C) during both childhood and young adulthood (P <.05). The positive association between parental CHD and LDL-C in young adulthood persisted independently of body mass index (BMI) and fasting insulin, but disappeared when fasting glucose was added to the model. With respect to triglycerides and VLDL-C, inclusion of BMI, insulin, and/or glucose eliminated the adverse association with parental CHD. These observations suggest that parental CHD is just one more explanatory variable that loses its partial contribution to lipoprotein profiles in their offspring when other strongly interrelated contributory variables such as age, body fatness, and measures of glucose homeostasis are taken into account. Information on these risk variables in conjunction with parental or family history of CHD may enhance the potential of CHD risk assessment in youth. PMID- 11735091 TI - Tumor necrosis factor alpha -238 and -308 polymorphisms do not associate with insulin resistance in hypertensive subjects. AB - It is well established that, as a group, patients with essential hypertension are characterized by insulin resistance. Previous studies have shown that a biallelic polymorphism in the tumor necrosis factor (TNF)alpha promoter position -308 and 238 might be involved in the insulin resistance state in diabetic and/or nondiabetic subjects. We determined these polymorphisms in 235 nondiabetic hypertensive subjects and 246 unrelated normotensive controls. Fasting plasma glucose, insulin, lipoprotein, leptin, and TNFalpha concentrations were measured, in addition to plasma glucose and insulin responses to a 75-g oral glucose tolerance test (OGTT). Insulin sensitivity was also determined by an insulin suppression test in 69 hypertensive and 76 normotensive individuals. The results showed no association of these genotypic distributions between hypertensive and normotensive individuals both at -308 (GG, GA, and AA were 80.9%, 17.9%, and 1.3% in hypertensives, 84.2%, 15.4%, and 0.4% in normotensives, chi(2) = 1.68, P =.432) and at -238 (GG, GA, and AA were 98.3%, 1.7%, and 0% in hypertensives, 96.7%, 3.3%, and 0% in normotensives, chi(2) = 1.19, P =.276) sites. These results did not change even after adjustment for values of age and body mass index (BMI). Anthropometric measurements, fasting plasma glucose, insulin, lipoprotein concentrations, glucose, and insulin responses to OGTT, TNFalpha, and leptin concentrations were similar between the genotype at the -308 site both in hypertensive and normotensive groups. Insulin sensitivity, either measured by an insulin suppression test or homeostasis model assessment (HOMA) index, did not differ between the genotype at the -308 site in subjects with hypertension or normotension. Fasting plasma TNFalpha (10.2 alpha 0.5 pg/mL v 10.1 +/- 0.5 pg/mL, P =.928) concentrations did not differ between hypertensive and normotensive subjects even after adjustment for body fat and BMI values. We conclude that TNFalpha promoter gene polymorphisms at position -238 and -308 do not play a major role in the pathogenesis of insulin resistance in Chinese subjects with or without hypertension. PMID- 11735092 TI - Decrease in the plasma von Willebrand factor concentration following glucose ingestion: the role of insulin sensitivity. AB - Elevated plasma von Willebrand factor (vWF) concentration is thought to be associated with increased prevalence of cardiovascular events in the insulin resistance syndrome. We examined the effects of oral glucose challenge and accompanying metabolic and hemodynamic changes on vWF levels with respect to insulin sensitivity. Forty normotensive and hypertensive subjects (mean age +/- SD, 40 +/- 5 years) underwent a standard oral glucose tolerance test (OGTT). Plasma vWF antigen, glucose, insulin, catecholamines, and hemodynamics were measured at rest, and at 30, 60, 90, and 120 minutes after glucose intake. Insulin sensitivity was determined by the insulin sensitivity index (ISI(0,120)). Resting plasma vWF concentration was associated with screening systolic blood pressure (BP) (r =.43, P =.005). There were time effects for all variables of interest. While vWF antigen (P =.044), epinephrine (P =.003), and diastolic BP (P =.001) decreased after glucose challenge, norepinephrine (P =.009), systolic BP (P =.022), and heart rate (P <.001) increased. Decline in vWF (area under the curve) was associated with decrease in epinephrine (r =.46, P =.004) and with screening systolic BP (r =.45, P =.004). However, neither resting plasma vWF levels nor vWF decrease following glucose ingestion were significantly associated with the ISI(0,120.) The plasma vWF concentration decreases following glucose ingestion. While mechanisms underlying this phenomenon may relate to sympathetic nervous system function, they seem not related to insulin sensitivity. Endothelial dysfunction such as caused by hypertension rather than metabolic dysregulation per se may underlie the elevated plasma vWF concentration found with insulin resistance. PMID- 11735093 TI - Beneficial effects of metformin in normoglycemic morbidly obese adolescents. AB - Hyperinsulinemia and insulin resistance are common features of obesity in humans and experimental animals. It has been demonstrated that metformin, an antihyperglycemic agent, decreases hyperinsulinemia and insulin resistance leading to decreased adiposity in obese and non-insulin-dependent diabetes mellitus (NIDDM) adults. To evaluate the antiobesity effect of metformin, we conducted a randomized double-blind placebo controlled trial in 24 hyperinsulinemic nondiabetic obese adolescents (body mass index [BMI] >30 kg/m(2)). All subjects were placed on a low-calorie (1,500 kcal for women and 1,800 kcal for men) meal plan. After an initial 1-week lead-in period, 12 subjects (mean +/- SE for age and BMI, 15.6 +/- 0.4 and 41.2 +/- 1.8, respectively) received metformin (850 mg twice daily) for 8 weeks, and 12 subjects (mean +/- SE for age and BMI, 15.7 +/- 0.5 and 40.8 +/- 1.4, respectively) received placebo. Compared to the placebo group, the metformin group had greater weight loss (6.5% +/- 0.8% v 3.8 +/- 0.4%, P <.01), greater decrease in body fat (P <.001), greater increase in fat-free mass to body fat ratio (P <.005), and greater attenuation of area under the curve (AUC) insulin response to an oral glucose tolerance test (P <.001). This was associated with enhanced insulin sensitivity, as determined by the fasting plasma glucose:insulin, 2-hour glucose:insulin, and AUC glucose:AUC insulin ratios, in the metformin group compared to controls (P <.01). This corresponded to a significant reduction in plasma leptin (P <.005), cholesterol, triglycerides, and free fatty acid (FFA) levels (P <.05) only in the metformin-treated subjects. Combined metformin treatment and low-calorie diet had a significant antiobesity effect in hyperinsulinemic obese adolescents compared to a low-calorie diet alone. PMID- 11735094 TI - Remnant-like particles cholesterol is higher in diabetic patients with coronary artery disease. AB - Diabetes mellitus (DM) has been well known to be one of the risk factors of coronary artery disease (CAD). Recently, remnant-like particles cholesterol (RLP C) has been reported to be associated with CAD. However, few studies reported the association of RLP-C level with CAD in subjects with DM. To investigate the effects of presence or absence of DM on the association between RLP-C and CAD, we compared the RLP-C level in 142 male patients with CAD and 123 male subjects without CAD (non-CAD), including 44 and 38 DM patients, respectively. RLP-C was significantly higher in CAD than non-CAD (P <.05). RLP-C and RLP-C/plasma triglyceride (TG) ratio in CAD with DM were higher than CAD without DM (P <.01, P <.05), and non-CAD with DM (P <.001, P <.05). There was positive correlation between RLP-C and plasma-TG in non-CAD without DM (r =.44, P <.01), non-CAD with DM (r =.56, P <.001), CAD without DM (r =.81, P <.0001), and CAD with DM (r =.75, P <.001). After excluding the hypertriglyceridemic patients (>200mg/dL), RLP C/plasma-TG ratio was significantly higher in CAD with DM than CAD without DM (P <.001) and non-CAD with DM (P <.05). These results suggest that increased RLP-C to plasma-TG may be associated with CAD in middle-aged diabetic male subjects. PMID- 11735095 TI - Hyperhomocysteinemia and related factors in 600 hospitalized elderly subjects. AB - Hyperhomocysteinemia (HHcy) is a metabolic disorder frequently occurring in the elderly population. Recently several reports have suggested abnormalities in homocysteine (tHcy) metabolism implicating HHcy as a metabolic link in the multifactorial processes characterizing many geriatric illnesses-with special emphasis on atherosclerotic vascular diseases and cognitive impairment. The present study was undertaken in a large sample of elderly hospitalized subjects to determine (1) the prevalence of HHcy, (2) the association of HHcy with vascular and cognitive disorders, and (3) the factors independently predicting Hhcy. Six hundred elderly subjects (264 men and 336 women; mean age, 79 +/- 9 years) were randomly chosen from those admitted as inpatients over a period of 3 years. In all patients, body mass index (BMI), mid-upper arm muscle area (MUAMA), plasma cholesterol, triglycerides, total proteins, albumin, lymphocyte count, creatinine, homocysteine (fasting and 4 hours after methionine oral load), serum vitamin B(6), vitamin B(12), and folate concentrations were measured. The presence of disease or use of medications known to affect homocysteine plasma levels were also recorded. The mean fasting tHcy level was 16.8 +/- 12 micromol/L in the whole sample, 18.18 +/- 13.25 micromol/L in men, and 15.86 +/- 12.14 micromol/L in women (P =.005 men v women). The mean Hcy level 4 hours after methionine load was 37.95 +/- 20.9 in the whole sample. Prevalence of hyperhomocysteinemia (fasting Hcy > or = 15 micromol/L or 4 hours after methionine load > or = 35 micromol/L) was 61% (365/600) (67% in men and 56% in women, P <.05). HHcy was rarely (8%) an isolated disorder; in addition to diabetes (20%), renal failure (48.2%), and malnutrition (20.2%), it was often associated with heart failure (30%), malignancies (20.5%), and the use of diuretics (56%) and anticonvulsant drugs (13%). Plasma homocysteine progressively increases across subjects from those with no diabetes, malnutrition, renal failure, obesity, inflammatory bowel disease, heart failure to those with 1, 2, or more concurrent diseases. Multiple stepwise regression analysis showed that 72% of plasma total fasting tHcy variability was explained by age, serum folate, plasma albumin, use of diuretics, and renal function (measured as plasma creatinine clearance). In conclusion, the present study documents that hyperhomocysteinemia, in elderly hospitalized patients is (1) a common finding, (2) frequently associated with vascular and cognitive disorders, and (3) probably a secondary phenomenon in most cases. The major predictor of high plasma homocysteine levels were age, serum folate, plasma albumin, plasma creatinine clearance, and use of diuretic drugs. These variables explain a large proportion of plasma Hcy variability. PMID- 11735096 TI - Serum phospholipid fatty acid composition and insulin action in type 2 diabetic patients. AB - Relationships have been demonstrated between insulin sensitivity and the fatty acid (FA) composition of serum and tissue lipids in adult humans. The present study aimed to investigate the above relationships in different groups of type 2 diabetic patients (DM2). The FA composition of serum phospholipids (S-PL) measured by gas liquid chromatography and insulin action during a 2-step hyperinsulinemic isoglycemic clamp (1 and 10 mU/kg. min) were determined in 21 newly diagnosed DM2 subjects (DMN), in groups of long-term DM2 patients treated with hypoglycemic agents (DMH; n = 21) or diet alone (DMD; n = 11), and in 24 healthy subjects (HS). In diabetics, the metabolic clearance rates of glucose at both insulin levels (MCR(glu)submax and MCR(glu)max) were significantly reduced compared with HS (MCR(glu)submax DMN, 5.35 +/- 2.7 mL x kg(-1) x min(-1), DMH, 5.38 +/- 2.17 mL x kg(-1) x min(-1); DMD, 5.48 +/- 2.35 mL x kg(-1) x min(-1) v HS, 10.9 +/- 3.3 mL x kg(-1) x min(-1); P <.01; MCR(glu)max DMN, 13.3 +/- 3.3 mL x kg(-1) x min(-1); DMH, 12.5 +/- 3.0 mL x kg(-1) x min(-1); DMD, 13.3 +/- 3.0 mL x kg(-1) x min(-1) v HS, 17.4 +/- 3.8 mL x kg(-1) x min(-1); P <.05). Increased contents of highly unsaturated n-6 family FA (P <.01), arachidonic acid in particular (DMN, 10.98% +/- 1.79%; DMD, 10.78% +/- 1.64%; DMH, 10.97% +/- 1.7% v HS, 8.51% +/- 1.53%; P <.001), were found in all groups of diabetics compared with HS, while lower levels of linoleic acid were seen in DMN (P <.001) and DMH (P <.05). The contents of saturated FA and monounsaturated FA were comparable in HS, DMN, and DMD. While in HS there were significant negative correlations between MCR(glu) and the contents of saturated FA and a positive association between insulin action and proportions of linoleic and arachidonic acids, no significant relationships were found in diabetic subjects. Different groups of DM2 patients show an altered FA pattern of S-PL, which is not related to insulin action. The above data support the hypothesis that changes in FA composition may play a role in modulating insulin action in peripheral tissues, but cannot explain the insulin resistance (IR) in DM2 patients. PMID- 11735097 TI - Circulating granulocyte-macrophage colony-stimulating factor and serum fatty acid composition in men and women. AB - Atherosclerosis is increasingly recognized as an inflammatory disease. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a proinflammatory cytokine, recently implicated as a prominent component of the regulatory network involved in atherogenesis. We aimed to study the relationship between circulating GM-CSF levels and serum fatty acid (FA) composition in 78 healthy subjects. The latter was analyzed by gas-liquid chromatography and GM-CSF by a high-sensitivity commercial enzyme-linked immunosorbent assay (ELISA). Among women (n = 40), serum GM-CSF levels were found to be positively associated with the proportion of palmitic acid (C16:0) and negatively with linoleic acid (C18:2omega-6), docosahexaenoic acid (DHA, C22:6omega-3), and the proportion of total essential FA. After excluding smoking women (n = 6), the associations among GM-CSF and serum linoleic acid concentration (r = -0.49, P =.003), arachidonic acid (r = 0.52, P =.001), and DHA (r = -0.34, P =.04) were strengthened. The ratio of palmitic to linoleic and DHA acids was the single best predictor of serum GM-CSF in all subjects. Together with arachidonic acid, it contributed to 22% of the GM CSF variance in women, after taking into account the effects of age, body mass index (BMI), blood pressure, and smoking status. None of these associations were observed among men. In conclusion, serum FA composition is associated with circulating GM-CSF specifically in women. As human arterial and venous smooth muscle cells release GM-CSF, and treatment of endothelial cells with oxidized low density lipoproteins results in a rapid expression of GM-CSF, the mechanisms involved in these associations and the sex-linked differences should be further explored. PMID- 11735098 TI - Exercise-induced oxidative stress in older adults as measured by antipyrine oxidation. AB - Aging is associated with increased susceptibility to free radical-mediated tissue damage. Measuring exercise-induced oxidative stress, however, is a major problem in free radical research. We used an exogenous marker (antipyrine) to measure oxidative stress in older adults during submaximal exercise. Antipyrine pharmacokinetics is independent of blood flow to the liver. Furthermore, antipyrine reacts quickly with hydroxyl radicals (10(10)-10(11) L x mol(-1) x s( 1)) to form para- and ortho-hydroxyantipyrine (o-APOH). o-APOH is not formed in man through the mono-oxygenase pathway of cytochrome P450. Thirty-four subjects (62 +/- 1 years) orally ingested 10 mg antipyrine/kg body mass. One hour after ingestion subjects cycled 45 minutes at 50% maximal power output. Exercise significantly increased the ratio of para-hydroxyantipyrine (p-APOH) to native antipyrine in plasma (.0014 +/-.0001 v.0021 +/-.0002; P <.0001). Also, the ratio of o-APOH was significantly increased after exercise (.0014 +/-.0001 v.0019 +/ .0002; P <.0001). Exercise significantly increased plasma levels of plasma malondialdehyde MDA) (.55 +/-.07 v.92 +/-.21 micromol/L; P <.01). In conclusion, in older adults, oxidative stress occurs during cycling at submaximal intensity as measured with free radical reaction products of antipyrine. PMID- 11735099 TI - Glucose and insulin regulate glycosylphosphatidylinositol-specific phospholipase D expression in islet beta cells. AB - Insulin resistance is associated with a compensatory islet hyperactivity to sustain adequate insulin biosynthesis and secretion to maintain near euglycemia. Both glucose and insulin are involved in regulating proteins required for insulin synthesis and secretion within the islet and islet hypertrophy. We have determined that glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is present within the secretory granules of islet beta cells. To determine if GPI PLD is regulated in islet beta cells, we examined the effect of glucose and insulin on GPI-PLD expression in rat islets and murine insulinoma cell lines. Glucose (16.7 mmol/L) increased cellular GPI-PLD activity and mRNA levels 2- to 7 fold in isolated rat islets and betaTC3 and betaTC6-F7 cells. Insulin (10(-7) mol/L) also increased GPI-PLD mRNA levels in rat islets and betaTC6-F7 cells 2- to 4-fold commensurate with an increase in GPI-PLD biosynthesis. To determine if islet GPI-PLD expression is increased in vivo under conditions of islet hyperactivity, we compared GPI-PLD mRNA levels in islets and liver from ob/ob mice and their lean littermates. Islet GPI-PLD mRNA was increased 5-fold while liver mRNA and serum GPI-PLD levels were reduced 30% in ob/ob mice compared with lean littermate controls. These results suggest that glucose and insulin regulate GPI-PLD mRNA levels in isolated islets and beta-cell lines. These regulators may also account for the increased expression of GPI-PLD mRNA in islets from ob/ob mice, a model of insulin resistance and islet hyperactivity. PMID- 11735100 TI - Effect of cold acclimation on brown adipose tissue fatty acid synthesis in rats adapted to a high-protein, carbohydrate-free diet. AB - The effect of cold acclimation on brown adipose tissue (BAT) fatty acid synthesis was investigated in rats adapted to a high-protein, carbohydrate-free diet. At an ambient temperature (25 degrees C), rates of fatty acid synthesis in BAT from rats adapted to the high-protein diet were reduced to 27% of rats fed the balanced diet and increased markedly after cold acclimation (10 days at 4 degrees C), although the increase was smaller than in control rats. BAT weight increase induced by cold acclimation was smaller in rats fed the high-protein diet (30%) than in controls (100%). When expressed per whole tissue, maximal activities of BAT glucose-6-phosphate dehydrogenase, malic enzyme, adenosine triphosphate (ATP) citrate lyase, and acetyl-coenzyme A carboxylase were markedly reduced in high protein diet-adapted rats at 25 degrees C and increased after cold acclimation in BAT from the 2 groups. However, when expressed per milligram protein, only acetyl coenzyme A carboxylase showed an increase in both controls and in rats fed the high-protein diet. G6P-dehydrogenase, malic enzyme, and ATP-citrate lyase increased (per milligram protein) only in rats adapted to the high-protein diet and actually decreased in BAT from cold-acclimated control rats. Initial (before activation) pyruvate dehydrogenase (PDH) complex activity was lower in BAT from rats fed the high-protein diet at 25 degrees C and increased in cold-acclimated rats from the 2 groups. Circulating levels of insulin decreased in the 2 groups after cold acclimation. The data suggest that the cold acclimation-induced increase in BAT lipogenesis in rats adapted to the high-protein diet was due to a restoration of sympathetic activity, which induced both BAT hyperplasia and activation of adipocyte free fatty acid (FFA) synthesis, with an important participation of acetyl-coenzyme A carboxylase and pyruvate dehydrogenase. PMID- 11735101 TI - Are there persons who are obese, but metabolically healthy? AB - The aim of this article was to review the evidence for a metabolically normal subset of the obese and its implications for clinical and research work. The methods included literature review and correspondence with authors. Since 1947, when Vague described a relation between distribution of body fat and the risk factors for cardiovascular disease, much evidence has suggested that early onset of the obesity, hyperplasia of normal adipocytes, and normal quantities of visceral abdominal fat may be associated with a favorable metabolic response in obese subjects. Analyses in 1973 by Keyes and later by Reuben Andres in 1980 suggested that obesity for some was not a risk factor and might even be an asset. Recently, in the study by Bonora et al of the relation between insulin resistance and the 4 main disorders of the metabolic syndrome in the Bruneck epidemiologic study, a subgroup of obese individuals with a normal metabolic response was evident. In a current study by Brochu et al of an obese metabolically normal subgroup of postmenopausal women, visceral abdominal fat estimated by computed tomography (CT) scan and age of onset were significant variables. The obese, metabolically normal subgroup (OBMN) must be taken into consideration in both clinical and research work. Persons with OBMN and their parents may be wrongly blamed because of the obesity. Attempts at weight loss may be counterproductive. The criteria for selection of obese research subjects may favor inclusion of an OBMN subset, which may invalidate statistical analysis. Findings suggesting the OBMN subset include family members with uncomplicated obesity, early onset of the obesity, fasting plasma insulin within normal range, and normal distribution of the excess fat. Hormonal, genetic studies, and prospective studies will help to clarify the significance and underlying mechanisms of this subset. PMID- 11735102 TI - The role of platelets in decrypting monocyte tissue factor. AB - Although about 80% of tissue factor (TF) extracellular domain antigen present in lipopolysaccharide (LPS)-stimulated monocytes is available at the cell surface, only 10% to 20% of the total extractable TF activity is expressed on the surface of intact monocytes. Thus, most of the TF activity is latent or encrypted in the cell membrane. When coincubated, leukocytes and platelets generate more TF activity than either cell type alone. We have shown that such platelet-promoted enhancement of LPS-induced TF activity in monocytes in whole blood depends on neutrophil involvement in a P-selectin/CD15 (a leukocyte membrane-bound carbohydrate)-dependent reaction. The effect was even more pronounced when both the phorbol ester, phorbol 12-myristate 13-acetate (PMA), and LPS were present during monocyte stimulation. We currently envisage that decryption is mediated through the secretion of TF-rich particles by monocytes. These particles express CD15 and bind P-selectin exposed on either activated platelets or platelet derived microparticles. Interactions and fusion events, that typically occur between monocytes and platelets, would facilitate the generation of monocytes/monocyte microparticle and platelets/platelet microparticle hybrids, leading to particles rich in decrypted TF activity. In conclusion, platelets play a pivotal role in decrypting TF activity of monocytes, generating a hybrid TF terrain, which both triggers and favors thrombogenesis. PMID- 11735103 TI - The action of high-dose factor VIIa (FVIIa) in a cell-based model of hemostasis. AB - We have developed a cell-based model of hemostasis. This model suggests that the defect in hemophilia is specifically a failure of platelet-surface factor Xa (FXa) generation, leading to a failure of platelet surface thrombin generation. Activation of FX by FVIIa/tissue factor (TF) does not compensate for a lack of FXa activation on the platelet surface by the FVIIIa/FIXa complex. This is because plasma protease inhibitors prevent FXa from moving through the fluid phase from the TF-bearing cell to the platelet surface. We have previously proposed a platelet-dependent mechanism of action for high-dose factor VIIa (FVIIa; Novoseven, Novo Nordisk, Copenhagen, Denmark). Our data suggest that, when present at high levels, FVIIa binds to activated platelets and activates small amounts of FX independent of TF. This platelet-surface FXa can partially restore platelet-surface thrombin generation in hemophilia. Recently, van't Veer and colleagues reported results from an in vitro model in which coagulation reactions were initiated by relipidated TF. The authors concluded that high-dose FVIIa may exert a hemostatic effect in hemophilia by overcoming inhibition of FVIIa/TF activity by zymogen FVII. By contrast, we found that plasma levels of FVII did not slow thrombin generation in a model system initiated with cell associated TF. This discrepancy highlights the potential differences between the studies of the coagulation reactions assembled on living cells compared to phospholipid vesicles. Our data suggest that in a cellular system high-dose FVIIa acts primarily by enhancing the rate of thrombin generation on platelet surfaces and not by overcoming inhibition by zymogen FVII of TF-dependent activation of FX. PMID- 11735104 TI - Recombinant factor VIIa (Novoseven) restores deficient coagulation: experience from an ex vivo model. AB - The action of recombinant factor VIIa (rFVIIa) in coagulation deficiencies with increased risk of bleeding was investigated using in vitro perfusion. Blood samples were drawn from healthy donors, a patient with hemophilia A and inhibitors, and six patients undergoing oral anticoagulant treatment. Fragmin 10 U/mL was used as anticoagulant. rFVIIa (10 microg/mL in plasma) was added to blood samples, incubated for 1 minute at 37 degrees C, and perfusion studies performed for 10 minutes at 600 x s(-1) through annular chambers containing damaged vascular segments. Subendothelial fibrin and platelets were expressed as a percentage of subendothelial surface screened. Under different conditions, rFVIIa consistently restored or improved fibrin formation on the damaged vascular subendothelium exposed to circulating blood. It restored fibrin deposition in blood from the hemophilia A patient; in patients undergoing acenocoumarol treatment, it reduced the international normalized ratio (INR) from 2.47 to 1.25 with a significant increase in fibrin deposition. Platelet deposition varied slightly between clinical conditions but was less evident in the hemophilia A patient. These data support the concept that rFVIIa facilitates fibrin formation in these clinical situations, promoting procoagulant activity at sites of vascular damage where tissue factor is exposed. This could improve hemostasis in patients with hemophilia A and inhibitors, and in patients treated with oral anticoagulants. PMID- 11735105 TI - Effect of recombinant factor VIIa (Novoseven) on thrombocytopenia-like conditions in vitro. AB - Recombinant factor VIIa (rFVIIa; NovoSeven, Novo Nordisk, Copenhagen, Denmark) may help to promote hemostasis in patients with thrombocytopenia. We used two in vitro models of thrombin generation to evaluate this effect. The reconstituted model contained tissue factor (TF)-expressing monocytes, unactivated platelets, isolated plasma coagulation proteins, and calcium. Platelet activation and thrombin generation were measured in timed aliquots. In the plasma-based model, thrombin generation was measured continuously after the addition of lipidated TF and calcium to platelet-rich plasma using a slowly cleaved fluorescent substrate. Thrombocytopenic conditions were mimicked by decreasing the platelet density. In both systems, a platelet density-dependent lowering of the thrombin-generation peak was observed. Addition of rFVIIa to samples with low platelet density (6700 to 10000/microL) increased the initial thrombin generation in both systems without normalizing thrombin-generation curves. The magnitude of the rFVIIa effect was most pronounced in the plasma-based model. Platelet activation was not significantly delayed at low platelet density in the reconstituted model. Addition of rFVIIa to samples with low platelet density caused faster platelet activation, most likely as a consequence of the increased initial thrombin generation. The data suggest that rFVIIa may help to achieve hemostasis at low platelet densities by increasing the initial thrombin generation, thereby compensating for low platelet number. PMID- 11735107 TI - Inhibitor treatment: state of the art. AB - The treatment of hemophilic patients with inhibitors presents many challenges. Recombinant activated factor VII (rFVIIa; NovoSeven, Novo Nordisk, Copenhagen, Denmark) is a new therapeutic option for this patient population. Optimization of dosing regimens may be achieved by considering (1) individual pharmacokinetics (especially in the pediatric population); (2) high-dose bolus regimens; (3) continuous infusion (CI) including high-dose CI; (4) early treatment with NovoSeven in the course of a bleeding event; and (5) use of NovoSeven as a prophylactic therapy. The literature involving these issues is reviewed here. Institution of early therapy has been shown to decrease total NovoSeven requirement while increasing efficacy. Pediatric patients may require a higher dose of NovoSeven due to the higher clearance rate and shorter half-life demonstrated by this population. Therefore, the patient's age and/or individual pharmacokinetics and clearance rate should be considered when calculating doses and CI rates. High-dose NovoSeven has been shown to be effective in some patient populations without increased reported adverse events. Despite its short half life, prophylactic use of NovoSeven has been reported in a small number of patients, with an apparent decrease in bleeding episodes experienced. There has been significant interest in administration of NovoSeven by CI, particularly in patients requiring prolonged treatment, and it has been shown to be successful, although the optimal target factor VII activity has yet to be established and some authors have recommended the use of concomitant antifibrinolytics. PMID- 11735106 TI - Use of recombinant factor VIIa (NovoSeven) in patients with Glanzmann thrombasthenia. AB - Recombinant factor VIIa (rFVIIa; NovoSeven, Novo Nordisk, Bagsvaerd, Denmark) appears effective and relatively safe for the treatment of bleeding and for surgical prophylaxis in patients with Glanzmann thrombasthenia as reported to the International Registry on rFVIIa and Congenital Platelet Disorders. One of the shortcomings of the Registry data is the heterogeneity of treatment protocol, including dosage, number of doses used, duration of treatment before declaration of failure, and mode of rFVIIa administration (bolus v continuous infusion). The data are not yet sufficient to define optimal regimens for various indications such as the type of bleeding or the type of procedures. The place of this drug compared to platelet transfusion in the overall management of patients with Glanzmann thrombasthenia will need to be determined in relationship to a number of challenges and unresolved issues in the clinical care of these patients. These issues include: how to improve local measures for patients with mucosal bleeds, optimal management of young women during menarche, optimal platelet transfusion regimens for various indications, the relationship between antiplatelet antibodies detected by monoclonal antibody-specific immobilization of platelet antigens (MAIPA) and effectiveness of platelet transfusion, whether there are other biological tests that may correlate with effectiveness of platelet transfusion, and management of pregnancy and delivery regarding antiplatelet immunization. PMID- 11735108 TI - Tissue factor in experimental acute lung injury. AB - Acute lung injury (ALI) is characterized by fibrin deposition in the tissue and vascular spaces. Coagulation is activated after exposure to endotoxin or bacteria, and a procoagulant environment rapidly develops in the vascular, interstitial, and alveolar spaces of the lung. These changes are tissue factor (TF)-dependent and associated with increases in inflammatory cytokines. Procoagulant changes also occur in the lungs of patients with the acute respiratory distress syndrome (ARDS), suggesting that epithelial inflammation activates the extrinsic pathway. Many inflammatory mediators have specific effects on coagulation; however, the role of TF in regulation of pulmonary inflammatory responses is less clear. Here we report initial data on blockade of TF-initiated coagulation in baboons with Escherichia coli sepsis-induced ALI, using active site-inactivated FVIIa (FVIIai ASIS). Treatment with FVIIai prevented plasma fibrinogen depletion and attenuated fibrin deposition in the tissues. The drug also decreased systemic cytokine responses and inflammatory changes in the lung, including neutrophil infiltration, and decreased edema. Coagulation blockade with FVIIai improved lung function by preserving gas exchange and compliance, decreased pulmonary hypertension, and enhanced renal function. These results show that TF-FVIIa complex is an important regulatory site for the pathologic response of the lung to sepsis. PMID- 11735109 TI - Active site-inhibited seven: mechanism of action including signal transduction. AB - Vascular injury brings tissue factor (TF) into contact with blood and its natural ligands, factors VII (FVII) and VIIa (FVIIa). This results in localized FVIIa activity on TF-expressing cells, initiating coagulation, and nonhemostatic activities. Activation of transcription factors, expression of genes for inflammation, tissue remodeling, and wound healing follow, but these mechanisms for maintaining vascular integrity may lead to pathophysiologic states. Recombinant FVIIa is converted into a catalytically inert protein by reactive site residues reacting with Phe-Phe-Arg-chloromethyl ketone. Active site inhibited FVIIa (ASIS) retains its affinity for TF and competes for FVIIa and FVII binding to TF, blocking FVIIa activity and FVII to FVIIa activation. It therefore acts as an antithrombotic agent and has been shown in previous studies on animal models of sepsis to prevent organ failure associated with fibrin deposition. Mitigation of inflammatory response and prolonged survival were remarkable and additional effects of TF blockage by ASIS not observed with inhibitors of downstream coagulation factors Xa and thrombin. This suggests that FVIIa/TF exerts a noncoagulopathic effect on cellular activities, attenuated by ASIS blocking FVIIa-induced signaling. The precise mechanism remains elusive but blockade of TF/FVIIa activity provides an attractive possibility for pharmaceutical intervention. In vitro measurements of ASIS-TF binding and FVIIa/TF inhibition are described, together with investigation of the FVIIa induced signaling pathway and gene expression. Additionally, possible implications of ASIS blockage for hemostatic and nonhemostatic aspects of the pathophysiology associated with vascular stress and injury are discussed. PMID- 11735110 TI - Recombinant factor VIIa (Novoseven) as a hemostatic agent. AB - Recombinant activated factor VII (rFVIIa; NovoSeven, Novo Nordisk, Denmark) induces hemostasis in life- and limb-threatening bleeds and in major surgery of hemophilia A and B patients, regardless of inhibitor titer. A total of more than 6,500 patients have been treated, and NovoSeven has been administered in more than 180,000 standard doses. Experience gained from these clinical situations suggests that NovoSeven should be administered as a 90- to 110-microg/kg bolus dose every second hour. Hemophilia patients with mild to moderate bleeding episodes require two to three doses to achieve complete hemostasis, whereas patients with severe bleeding episodes may require more doses. For major surgery and in cases of life-threatening bleeding, dosing every second hour for the first 24 hours may be required. Thereafter, the same dose, but with longer intervals between doses, is recommended. Recent in vitro experiments indicate that even higher doses of NovoSeven may be needed to achieve full thrombin generation in the absence of factor VIII (FVIII), factor IX (FIX), and factor XI (FXI). PMID- 11735111 TI - Recombinant factor VIIa (Novoseven) and the safety of treatment. AB - Recombinant factor VIIa (rFVIIa; NovoSeven), Novo Nordisk, Bagsvaerd, Denmark) has been used to treat more than 6,500 patients with hemophilia or other bleeding disorders, and by May 2001, more than 180,000 standard doses had been administered. Clinical experience to date suggests that rFVIIa is safe and effective, with a very low incidence of thrombotic events being reported. Moreover, in the majority of cases, thrombotic events appear to be caused by improvements in the hemostatic system, rather than a direct effect of the agent itself. PMID- 11735112 TI - Alternative splicing of caspase-8 mRNA during differentiation of human leukocytes. AB - Caspase-8 is a key initiator of death receptor-induced apoptosis. Here we provide evidence that caspase-8 expression is subject to posttranscriptional regulation in human leukocytes. Resting peripheral blood lymphocytes preferentially use a distant splice donor site at the 3'-end of caspase-8 exon 8 to generate mRNAs with a truncated open reading frame. When lymphocytes were activated, the expression of caspase-8 variants was shifted to caspase-8/a and b which lack the extension of exon 8. The opposite change of the splicing pattern was found in a neutrophil differentiation model. Promyelocytic HL-60 cells mainly expressed caspase-8 mRNAs with the normal exon 8, but the splicing pattern was changed to the distant exon 8 splice site during DMSO-induced differentiation of HL-60 cells. In spite of the presence of these novel mRNAs, the corresponding translation products were not detectable in either cell type. Our findings suggest that leukocyte differentiation and alternative splicing of caspase-8 pre mRNA are inter-dependent processes. PMID- 11735113 TI - CD9 amino acids critical for upregulation of diphtheria toxin binding. AB - CD9 associates with a diphtheria toxin receptor (DTR) that is identical to the membrane-anchored form of heparin-binding EGF-like growth factor. We determined the region of CD9 important for upregulation activity. Human and monkey CD9 upregulates DT binding activity of DTR, while mouse CD9 has no upregulation activity. Transfection of chimeric constructs comprising monkey and mouse CD9s showed that the human sequence between Ala156 and Asp183 is essential for the upregulation activity. Studies of mutants, replacing a single amino acid within the region between Ala156 and Asp183 of monkey CD9 with the corresponding amino acid residue in mouse CD9, revealed that substitution of Gly158 is critical for the reduction of the upregulation activity and secondly for the substitution of Val159 and Thr175. These three amino acid residues were deduced to be located on the head domain of the second extracellular loop, suggesting that interactions of CD9 with DTR or DT at the domain containing these three amino acids were important for the upregulation of DT binding. PMID- 11735114 TI - Mice lacking Ca(v)2.3 (alpha1E) calcium channel exhibit hyperglycemia. AB - To investigate the functional role of Ca(v)2.3 channel in glucose homeostasis, we performed in vivo glucose tolerance and insulin tolerance tests together with stress-induced glucose release tests using mice deficient in Ca(v)2.3 channel (Ca(v)2.3-/-). The Ca(v)2.3-/- mice were significantly heavier than wild-type mice. In glucose tolerance and insulin tolerance tests, Ca(v)2.3-/- mice showed a significantly higher blood glucose level compared to wild-type mice. However, stress-induced blood glucose changes in Ca(v)2.3-/- mice were similar to those in wild-type mice. These results suggest that Ca(v)2.3 channel plays a role in glucose homeostasis by reducing insulin sensitivity and that Ca(v)2.3-/- mice exhibit symptoms resembling non-insulin-dependent diabetes mellitus. PMID- 11735115 TI - Role of ERK1/2 in the differential synthesis of progesterone and estradiol by granulosa cells. AB - A major concept in mammalian ovarian physiology is that follicle-stimulating hormone (FSH) activates the granulosa cells (GCs) in the Graafian follicle to selectively produce estradiol, but not progesterone, during the follicular phase of the menstrual or estrous cycle. However, given the fact that FSH can induce production of both estradiol and progesterone by GCs cultured in vitro, it has been postulated for a long time that there is a factor present in the ovary that selectively prevents FSH-induced progesterone production. Here, we provide evidence that two members of the mitogen-activated protein kinase family, extracellular signal-regulated kinase-1 and -2 (ERK1/2) can differentially regulate FSH-stimulated estradiol and progesterone production. Using primary rat GCs from early antral follicles cultured in serum-free medium for 48 h, we found that the addition of a specific inhibitor of ERK1/2 activation, U0126, caused the attenuation or enhancement of FSH-induced progesterone or estradiol production, respectively, in a dose-dependent manner. Throughout the 48-h culture period in this culture system ERK1/2 molecules in their activated state (phospho-ERK1/2) were clearly detectable in GCs exposed to FSH. The addition of U0126 caused a decrease in the levels of phosphorylated but not unphosphorylated ERK1/2 which was maintained throughout the 48-h culture, suggesting that U0126 was continuously active to inhibit the phosphorylation of ERK1/2. The divergent regulation of FSH-induced progesterone and estradiol synthesis by U0126 was further supported by demonstrating that U0126 inhibits and stimulates FSH-induced mRNA levels of steroidogenic acute regulatory protein and P450 aromatase, respectively. Collectively, this study clearly identified ERK1/2 as the first intracellular signaling molecules that differentially regulate FSH-induced progesterone and estradiol synthesis in GCs. PMID- 11735116 TI - Signals via FGF receptor 2 regulate migration of endothelial cells. AB - Fibroblast growth factors (FGFs) stimulate angiogenesis, of which signals are transduced via FGF receptor (FGFR) tyrosine kinases. Although FGFR1 is a major receptor in endothelial cells, FGFR2 is frequently detectable in endothelial cells. We have previously demonstrated that the intracellular domain of FGFR1 sufficiently transduced signals leading to proliferation, migration, urokinase secretion, and tube formation. However, little is known about the roles of signaling via FGFR2 alone in endothelial cells. Murine brain capillary endothelial cells, denoted IBE cells, express small amounts of IIIc FGFR2, which is not activated by keratinocyte growth factor (KGF). We then transfected the IIIb FGFR2 in these cells. Three stable cell lines expressing IIIb FGFR2 demonstrated chemotaxis toward KGF, but never proliferated, secreted urokinase, or formed tube-like structure by KGF treatment. Weak but sustained activation of mitogen-activated protein kinase (MAPK) was observed in these cells. Chemotaxis toward KGF was significantly attenuated by treatment with PD98059. This is the first demonstration that signaling solely via FGFR2 in endothelial cells only contributes to motility through MAPK. PMID- 11735117 TI - A secretion signal is present in the Collybia velutipes oxalate decarboxylase gene. AB - The oxalate decarboxylase (OXDC) gene from Collybia velutipes is overexpressed as an active form in Schizosaccharomyces pombe. The recombinant enzyme shows similar pH optima and stability, while substrate kinetic analysis shows a ninefold decrease in K(m) value with respect to native OXDC. Most of the expressed protein was present in periplasm and remained firmly bound to cell-wall materials. However, 20% of enzyme expressed was secreted out into the medium suggesting the presence of a secretion signal (C. velutipes) in the oxalate decarboxylase gene. This secretion signal is associated with the N-terminal of OXDC as is evident by secretion of nonsecretory genes AmA1 and beta-galactosidase. An expression vector using this signal is constructed for expression and secretion of heterologous proteins in S. pombe. PMID- 11735118 TI - Proton exchange as a relaxation mechanism for T1 in the rotating frame in native and immobilized protein solutions. AB - T1 relaxation in the rotating frame (T1rho) is a sensitive magnetic resonance imaging (MRI) contrast for acute brain insults. Biophysical mechanisms affecting T1rho relaxation rate (R1rho) and R1rho dispersion (dependency of R1rho on the spin-lock field) were studied in protein solutions by varying their chemical environment and pH in native, heat-denatured, and glutaraldehyde (GA) cross linked samples. Low pH strongly reduced R1rho in heat-denatured phantoms displaying proton resonances from a number of side-chain chemical groups in high resolution 1H NMR spectra. At pH of 5.5, R1rho dispersion was completely absent. In contrast, in the GA-treated phantoms with very few NMR visible side chain groups, acidic pH showed virtually no effect on R1rho. The present data point to a crucial role of proton exchange on R1rho and R1rho dispersion in immobilized protein solution mimicking tissue relaxation properties. PMID- 11735119 TI - An immunoreactive peptide of the FSH involved in autoimmune infertility. AB - The purpose of this study was to identify autoantigens contained in human ovary extracts. Serum samples from 36 infertile women with anti-ovary antibodies as detected with an ELISA technique were tested in Western blot against human ovary extracts. A reactive protein with a molecular mass matching that of the FSH was detected in 34 cases. These serum samples also reacted strongly in Western blot and ELISA with purified FSH and, in immunofluorescence, with pituitary cells. Using the Pepscan approach, with overlapping peptides matching the amino acid sequence of the human FSH beta-chain, several immunoreactive regions were evidenced. The 78-93 amino acid sequence of the human FSH beta-chain appeared as one of the major epitopes. Synthetic peptides of this region were prepared and demonstrated to react with human serum samples from women with anti-ovary antibodies. These data demonstrate that FSH can be an autoantigen, recognized by autoantibodies associated with infertility. PMID- 11735120 TI - Loss of a glycine in the alpha2 domain affects MHC peptide binding but not chaperone binding. AB - Prior to the binding of peptide in the endoplasmic reticulum (ER), the major histocompatibility complex (MHC) class I heavy chain associates with an assembly complex that includes the transporter associated with antigen processing (TAP). The proximity of a part of the MHC class I alpha2 domain alpha-helix to areas previously shown to influence assembly complex binding suggests that this region might also be involved in chaperone association. Position 151, found in this part of the alpha2 domain alpha-helix, has a side chain that points up, away from direct contact with peptide, and is occupied by a glycine in all murine MHC class I heavy chains. We found that substitution of this glycine in H-2L(d) with a histidine substantially increased the proportion of peptide-free forms, although TAP binding was not abrogated. Thus, interaction of the heavy chain with peptides, but not with the assembly complex, is influenced by this glycine. PMID- 11735121 TI - Kinetic determinants of the interaction of enoyl-ACP reductase from Plasmodium falciparum with its substrates and inhibitors. AB - We have recently demonstrated that Plasmodium falciparum, unlike its human host, has the type II fatty acid synthase, in which steps of fatty acid biosynthesis are catalyzed by independent enzymes. This difference could be successfully exploited in the design of drugs specifically targeted at the different enzymes of this pathway in P. falciparum, without affecting the corresponding enzymes in humans. The importance of enoyl-ACP reductase (FabI) in the fatty acid biosynthesis pathway makes it an important target in antimalarial therapy. We report here the initial characterization of Plasmodium FabI expressed in Escherichia coli. The K(m) values of the enzyme for crotonyl-CoA and NADH were derived as 165 and 33 microM, respectively. Triclosan shows competitive kinetics with respect to NADH but is uncompetitive with respect to NAD(+), which shows that the binding of triclosan to the enzyme is facilitated in the presence of NAD(+). PMID- 11735123 TI - Galectin-3 mediates the endocytosis of beta-1 integrins by breast carcinoma cells. AB - Galectin-3, a beta-galactoside binding lectin, has been demonstrated to play a key role(s) in cell to extracellular matrix interaction. The precise mechanism by which it modulates cellular adhesion is presently unclear and warrants further studies. We hereby report that galectin-3 mediates the endocytosis of beta-1 integrins in a lactose-dependent manner. Interestingly we observed that galectin 3 was also rapidly internalized by the cells via the same pathway and the internalization was completely blocked by lactose. The endocytosis process was temperature dependent and was inhibited by filipin but not chlorpromazine. The endocytosis of galectin-3 and beta-1 integrins by the cells was accompanied by rapid cell spreading due to cytoskeletal reorganization. The data suggest a novel mechanism by which galectin-3 and beta-1 integrins are internalized into breast carcinoma cells via a cavaleolae-like pathway of endocytosis. PMID- 11735122 TI - HMG-CoA reductase inhibitors reduce adhesion of human monocytes to endothelial cells. AB - HMG-CoA reductase inhibitors (statins) are believed to reduce coronary heart disease by mechanisms in addition to their well-known cholesterol-lowering effect. We studied the effect of these drugs on monocyte cell adhesion to endothelium. Pretreatment of monocytic cells (U937, THP-1, human CD14(+) monocytes) with 0.01-10 microM concentrations of atorvastatin, cerivastatin, or simvastatin significantly reduced cell adhesion to endothelium. In contrast, pretreatment of endothelium with statins did not affect adhesion of monocytes. Adhesion of monocytes to vascular cell adhesion molecule-1-coated dishes was reduced by these drugs. Cerivastatin also reduced PMA induction of NF-kappaB. Since monocyte adhesion to endothelium is an early event in atherogenesis, treatment with statins in prevention of coronary heart disease may have additional salutary effects to lowering of plasma LDL cholesterol. Our results indicate that the reduction of monocyte adhesion by HMG-CoA reductase inhibitors may be considered as a class effect. PMID- 11735124 TI - NF-kappaB activation for constitutive expression of VCAM-1 and ICAM-1 on B lymphocytes and plasma cells. AB - Cytokine stimulation can activate NF-kappaB that triggers inducible expression of E-selectin, VCAM-1 (Vascular Cell Adhesion Molecule-1) and ICAM-1 (Intercellular Cell Adhesion Molecule-1) in endothelial cells. In the previous study, we have shown that B lymphocytes and plasma cells can express E-selectin by constitutive activation of NF-kappaB. Here we show that human B lymphocytes and ARH-77 plasma cells expressed VCAM-1 and ICAM-1 in a cytokine dispensable mechanism. NF-kappaB antagonists could inhibit their expressions in ARH-77 cells. The activities of NF kappaB for VCAM-1 and ICAM-1 promoters prior to cytokine stimulation were detected in ARH-77 cells using electrophoretic mobility shift assays. Again, NF kappaB antagonists could abrogate these promoter activities. Taken together, our results demonstrate that NF-kappaB activation is the underlying molecular mechanism for constitutive expression of E-selectin, VCAM-1, and ICAM-1 on human B lymphocytes and plasma cells. PMID- 11735126 TI - Dnmt3b, de novo DNA methyltransferase, interacts with SUMO-1 and Ubc9 through its N-terminal region and is subject to modification by SUMO-1. AB - Dnmt3b, a DNA methyltransferase, is essential for mammalian development potentially through its transcription repression activity. To comprehend the underlying regulatory mechanism of Dnmt3b, we isolated small ubiquitin-like modifier 1 (SUMO-1) and Ubc9 as Dnmt3b-interacting proteins using yeast two hybrid screens. Deletion analysis and colocalization experiment demonstrated that Dnmt3b interacts with SUMO-1 and Ubc9 at its N-terminal region. We also confirmed the modification of Dnmt3b by SUMO-1 in vivo. These results suggest that sumoylation may constitute a regulation mechanism of Dnmt3b in vivo. PMID- 11735125 TI - Inhibition of LOX-1 by statins may relate to upregulation of eNOS. AB - LOX-1, a receptor for oxidized low-density lipoprotein (ox-LDL), plays a critical role in endothelial dysfunction and atherosclerosis; both of these conditions are associated with diminished expression of constitutive endothelial nitric oxide synthase (eNOS). Recent studies show that HMG CoA reductase inhibitors (statins) exert cardioprotective effect. We examined the role of LOX-1 in eNOS expression and modulation of this relationship by two different statins, simvastatin and atorvastatin in human coronary artery endothelial cells (HCAECs). Ox-LDL (40 microg/ml) upregulated the expression of LOX-1; simultaneously, there was a reduction in eNOS expression. Pretreatment of HCAECs with simvastatin or atorvastatin (1 and 10 microM) reduced ox-LDL-induced upregulation of LOX-1 and downregulation of eNOS (both P < 0.05). High concentration of statins (10 microM) was more potent than the low concentration (1 microM) (P < 0.05). Both statins also attenuated ox-LDL-mediated activation of MAP kinase. These observations indicate that statins attenuate the effect of ox-LDL on eNOS expression. Inhibitory effect on LOX-1 and subsequently MAP kinase activity provides a potential mechanism of beneficial effects of statins beyond lowering cholesterol. PMID- 11735127 TI - Different suicide inactivation processes for the peroxidase and cyclooxygenase activities of prostaglandin endoperoxide H synthase-1. AB - Prostaglandin endoperoxide H synthases (PGHSs)-1 and -2 have a cyclooxygenase (COX) activity involved in forming prostaglandin G2 (PGG2) from arachidonic acid and an associated peroxidase (POX) activity that reduces PGG2 to PGH2. Suicide inactivation processes are observed for both POX and COX reactions. Here we report COX reaction conditions for PGHS-1 under which complete COX inactivation occurs but with > or = 60% retention of POX activity. The rates of POX inactivation were compared for native oPGHS-1 versus Y385F oPGHS-1, a mutant that cannot form the Tyr385 radical of COX Intermediate II; the rates were the same for both native and Y385F oPGHS-1. Our data indicate that a COX Intermediate II/acyl or product complex is the precursor in COX inactivation. However, another species, probably an Intermediate II-like species but with a radical centered on a tyrosine other than Tyr385, is the immediate precursor for POX inactivation. PMID- 11735128 TI - Differential expression of the L-plastin gene in human colorectal cancer progression and metastasis. AB - To identify molecular alterations in the progression of colorectal carcinoma, we analyzed gene expression profiles of colon cancer cell lines derived from primary and metastatic tumors from a single patient. Of 2280 cDNAs investigated using our in-house microarray, the expression of 6 genes (tumor-associated antigen L6, L plastin, the human homologue of yeast ribosomal protein S28, the B-cell translocation gene, mitochondrial aspartate-aminotransferase, and HLA-A) increased, while that of 2 genes (keratin 5 and phosphoglucomutase) decreased in metastatic-tumor-derived cells compared with primary-tumor-derived cells. Of these genes, we assessed the L-plastin gene, an actin-bundling protein, at the protein level using a tissue microarray consisting of 58 clinically stratified colorectal cancer specimens. Consistent with our microarray results, the expression of L-plastin was significantly correlated with the progression of cancer staging. Therefore, our results suggest that the L-plastin gene is a potential metastatic marker. In addition, combining cDNA microarrays and tissue arrays, as shown here, is thought to facilitate the rapid characterization of candidate biomarkers. PMID- 11735129 TI - Molecular cloning and characterization of mouse cardiac junctate isoforms. AB - Junctate is a newly identified integral ER/SR membrane calcium binding protein, which is an alternative splicing form of the same gene generating aspartyl beta hydroxylase and junctin. Screening a mouse heart cDNA library using canine junctin cDNA as a probe yielded three complete mouse heart cDNAs. One of the cDNAs is homologous to the previously reported human junctate. The three mouse junctate proteins are composed of 270, 259, and 215 amino acids (we named them junctate-1, -2, and -3). The apparent molecular masses of the mouse junctates in SDS-PAGE were in the range between 40 and 53 kDa. Northern and Western blot analyses indicate that mouse junctates are expressed in heart, brain, spleen, lung, liver, kidney, and stomach, but not in skeletal muscle. The apparent molecular weights of junctates from heart and brain were somewhat different from those from the other tissues tested, suggesting that there are tissue-specific expression patterns of the different junctate isoforms. Immunohistochemical studies showed that junctates were expressed both in ventricular and atrial tissues. This is the first study that shows the presence of 3 distinct cardiac junctate isoforms expressed in various mammalian tissues. PMID- 11735130 TI - A novel spermatogenesis-related factor-1 gene expressed in maturing rat testis. AB - A rat gene with testis-specific expression coinciding with spermatogenesis was cloned by differential display. This spermatogenesis-related factor-1 (SRF-1) gene was not expressed in other organs. Testicular expression was detected from 5 weeks of age and increased up to 15 weeks; this level of expression was maintained for 63 weeks. The 750-bp cloned gene was coded for an open reading frame of 202 amino acids. According to in situ hybridization at 7 weeks, this gene was expressed mainly in spermatocyte. The gene product may function as a molecular motor in meiosis, as the deduced amino acid sequence showed partial homology with kinesin-related proteins. The action of this gene and its product with respect to division of reproductive cells requires further investigation. PMID- 11735131 TI - Hypochlorite-modified (lipo)proteins are present in rabbit lesions in response to dietary cholesterol. AB - Myeloperoxidase (MPO), a heme enzyme secreted by activated phagocytes, generates an array of oxidants proposed to play critical roles in host defense, tissues damage, and foam cell formation. Although neutrophils are the major source for MPO, the enzyme could be identified abundantly in circulating monocytes and monocytes/macrophages in rabbit lesions. MPO is the only enzyme known to generate hypochlorous acid (HOCl) and HOCl-modified lipoproteins have pronounced atherogenic and/or proinflammatory features in vivo and in vitro. Using specific monoclonal antibodies, HOCl-modified (lipo)proteins were detected in atherosclerotic plaques of heterozygous Watanabe heritable hyperlipidemic rabbits and to a lesser extent in a specific strain of New Zealand White rabbits with a high atherosclerotic response to hypercholesterolemia. Colocalization of immunoreactive MPO and HOCl-modified-epitopes in serial sections of rabbit lesions provides convincing evidence for MPO-H2O2-chloride system-mediated oxidation of (lipo)proteins under in vivo conditions. We propose that monocyte derived MPO could connect chronic inflammatory conditions with arterial lipid/lipoprotein deposition during diet-induced atherogenesis in rabbits. PMID- 11735132 TI - Reactive oxygen species in mechanical stress-induced cardiac hypertrophy. AB - Mechanical stress induces various hypertrophic responses including activation of mitogen-activated protein kinases (MAPKs) in cardiac myocytes. Here we examined the role of the small GTP-binding proteins of Rho family and reactive oxygen species (ROS) in stretch-induced activation of p38MAPK in cardiomyocytes. Overexpression of dominant-negative mutants of Rac1 (D.N. Rac1), D.N.RhoA and D.N.Cdc42 suppressed stretch-induced activation of p38MAPK. Overexpression of constitutively active mutants of Rac1 (C.A.Rac1) and C.A.Cdc42 increased the p38MAPK activity in the absence of mechanical stress. Pretreatment with N-acetyl L-cysteine and N-(2-mercaptopropionyl)-glycine (NAC) suppressed stretch-induced activation of p38MAPK. Mechanical stretch increased intracellular ROS generation, which was abrogated by overexpression of D.N.Rac1 and attenuated by overexpression of D.N.RhoA and D.N.Cdc42. An increase in protein synthesis evoked by mechanical stretch was suppressed by overexpression of D.N.Rac1 and pretreatment with NAC. These results suggest that mechanical stress induces cardiac hypertrophy through the Rac1-ROS-p38MAPK pathway in cardiac myocytes. PMID- 11735133 TI - The chaperonins of Synechocystis PCC 6803 differ in heat inducibility and chaperone activity. AB - The chaperonins GroEL and Cpn60 were isolated from the cyanobacterium Synechocystis PCC 6803 and characterized. In cells grown under optimal conditions their ratio was about one to one. However, the amount of GroEL increased considerably more than that of Cpn60 in response to heat stress. The labile chaperonin oligomer required stabilization by MgATP or glycerol during isolation. Use of the E. coli mutant strain, groEL44 revealed that the functional properties of the two cyanobacterial chaperonins are strikingly different. Overexpression of cyanobacterial GroEL in the E. coli mutant strain allowed growth at elevated temperature, the formation of mature bacteriophage T4, and active Rubisco enzyme assembly. In contrast, Cpn60 partially complemented the temperature-sensitive phenotype, the Rubisco assembly defect and did not promote the growth of the bacteriophage T4. The difference in chaperone activity of the two cyanobacterial chaperonins very probably reflects the unique chaperonin properties required during the life of Synechocystis PCC 6803. PMID- 11735134 TI - Evidence for imaging the mandible in the management of oral squamous cell carcinoma: a review. AB - The accuracy of clinical examination, peroperative periosteal stripping and imaging techniques in predicting tumour invasion of the mandible in oral cancer so far reported have been compared according to their specificity and sensitivity. Clinical examination alone was not shown to be accurate, but periosteal stripping at the time of resection was extremely accurate although so far only one study has investigated it. No single imaging technique will accurately predict the invasion of tumour into the mandible, but a combination of an orthopantomogram and bone scintigraphy is recommended in early invasion. Magnetic resonance imaging is more sensitive than computed tomography and therefore may be more useful if mandibular invasion requires consideration. The decision to resect the mandible as part of the management of oral cancer should be taken on the evidence of clinical examination, periosteal stripping and at least two imaging techniques that complement each other in terms of specificity and sensitivity. PMID- 11735135 TI - Nerve morbidity following wisdom tooth removal under local and general anaesthesia. AB - Two hundred and one patients had unilateral removal of the lower third molars under local anaesthesia and a further 234 patients had either bilateral or unilateral removal under general anaesthesia. A total of 634 lower third molars were extracted by four experienced surgeons (two consultants and two senior grade staff). All patients were reviewed independently 1-week postoperatively and any sensory disturbance and its location was recorded. Patients with sensory disturbance were subsequently reviewed at 1 month and again at 6 months if recovery was not complete. This study demonstrated little difference in the adverse event rate per tooth extracted between procedures under local and general anaesthesia. However, within the general anaesthetic group, the few unilateral procedures showed evidence of higher risk, but the number was too small for valid satisfactory analysis. The risk of nerve morbidity was also greater where the duration of the procedure was longer than 15 minutes in unilateral cases. PMID- 11735136 TI - Evaluation of factors predictive of lingual nerve injury in third molar surgery. AB - The aim of this study was to investigate risk factors for temporary and permanent lingual nerve injury after extraction of mandibular third molars. It was based on a 4-year prospective study of 2134 consecutive mandibular third molar operations in 1384 consecutive day case patients. During the study period (1994-1998) data were collected prospectively on patient, dental and surgical factors and correlated with lingual nerve injury using Student's t test, xi(2) and multiple logistic regression analysis. The incidence of temporary and permanent lingual nerve injury was 1 and 0.3%, respectively, per tooth. Factors that predicted temporary and permanent lingual nerve injury by univariate analysis were age, depth of application, difficulty of operation, surgeon and surgical technique used. Independent risk factors identified by multivariate analysis for temporary lingual nerve injury were perforation of the lingual plate, exposure of the nerve and increased difficulty of operation. The predictors for permanent lingual nerve injury in order of importance were perforation of the lingual plate, surgeon, increased difficulty of operation, exposure of the nerve and increased age of the patient. Surgical factors are the main contributors to lingual nerve injury during third molar extraction, but patient and dental factors are also involved. PMID- 11735137 TI - Changes in occlusal state of patients with mandibular prognathism after orthognathic surgery: a pilot study. AB - This study examined the occlusal state of patients with mandibular prognathism and compared it with that of adults with normal occlusion (controls). It also examined changes in occlusal state after orthognathic operations in these patients. The values of occlusal contact area and bite force in patients before operation were significantly lower than in controls, and occlusal pressure in patients was higher than in controls. The occlusal contact area and bite force of the patients 1 month after the operation had decreased to below preoperative values. These values 12 months after the operation had increased by 2.0 and 1.8 times in women and 1.4 and 1.4 times in men, respectively, compared with preoperative values. However, absolute values remained extremely low compared with those of controls. In contrast to the above, occlusal pressure reached its maximum value 1 month after the operation and at 12 months it was close to the value for controls. PMID- 11735138 TI - Influence of routine bone grafting on the stability of the non-cleft Le Fort 1 osteotomy. AB - Although bone grafting is known to improve stability after midface cleft osteotomy, it is advocated only rarely for people without clefts. To assess the influence of routine bone grafting on stability in such patients we analysed radiographic data of 112 patients retrospectively. All patients had had Le Fort 1 osteotomy with autogenous bone grafts harvested by trephine from the iliac crest, and 76 had had concurrent mandibular procedures. Rigid internal fixation was adopted throughout. Impaction proved to be the most stable move. While there was little postoperative relapse of the anterior maxilla (0.9%), the posterior maxilla tended to move further upwards with time (2.3%). Overall relapse after advancement and inferior repositioning was low (3.5% and 5%, respectively). The only complication noted at the iliac crest donor site was spread of the scar, which affected three patients. When we compared these results with previously published data, they suggested that in patients without clefts rigid internal fixation combined with bone grafting may improve stability after selected Le Fort 1 osteotomy moves. PMID- 11735139 TI - Histomorphometric and densitometric changes in bone volume and structure after avascular bone grafting in the extremely atrophic maxilla. AB - AIM: To evaluate changes in volume and structure of bone after autogenous iliac crest bone grafting to the maxilla. To discover the predictive value of computed tomographic (CT) measurements of bone density and study their correlation with the results of histomorphometric analyses of bone structure. PATIENTS AND METHODS: In 25 patients with atrophic maxillae (mean age 47 years, range 15-71), who had onlay bone grafting and sinus floor augmentation with autogenous iliac crest bone, a CT analysis was carried out immediately before and after grafting, and 5 months (range 4-6) later, at the time of endosseous implantation. On both occasions, bone biopsy specimens were taken. HISTOLOGY OF BONE: After preparing and digitizing semithin sections, histomorphometry (NH-Image) was done to establish the ratio of trabeculae to medullary cavity. CT ANALYSIS: The available transverse and vertical bone and the bone density of natural and augmented bone were analyzed using the Simplant Program and the CT data. As reference, the bone density of the first cervical vertebra was used. By regression analysis, the correlation of CT bone density and histological bone structure was investigated. RESULTS: After a median healing period of 4.5 months, no significant changes were found in bone volume compared with the measurements made immediately after grafting. The changes in density in the cancellous portion of the grafted bone showed no correlation (correlation coefficient: +0.16) with the results of histomorphometric analysis of bone structure. CONCLUSIONS: The predictive value of measurement of bone density to evaluate bone structure on the basis of the D1 D4 classification needs to be reconsidered. The most favourable time for secondary implantation is 4-6 months after iliac crest bone grafting. PMID- 11735140 TI - An ergonomic and effective parotid retractor. PMID- 11735141 TI - Bone injector in sinus lift surgery. PMID- 11735142 TI - Bone regeneration using recombinant human bone morphogenetic protein-2 (rhBMP-2) in alveolar defects of primate mandibles. AB - The efficacy of bone morphogenetic protein (BMP) for bone reconstruction has been widely studied in numerous animal experiments, but insufficient information exists about its ability to regenerate bone in primates. The purpose of this study was to evaluate the effects of recombinant human BMP-2 (rhBMP-2) on bone formation in alveolar bone defects in the mandibles of young primates. Marginal bone defects were created in the mandibles of nine 5-year-old rhesus monkeys and rhBMP-2 permeated in a polylactic-co-glycolic acid-coated gelatin sponge (PGS) was implanted into the bone defects. The resected bone treated with rhBMP-2 regenerated completely at 12 weeks postoperatively, and remodelling and consolidation of new bone were seen histologically. This study provides evidence of considerable bone regeneration in alveolar defects after surgical implantation of rhBMP-2 in non-human primates. This technique may be an effective alternative to autogenous bone grafts for reconstructive surgery in clinical practice. PMID- 11735143 TI - Changing incidence of oral and maxillofacial tumours in East Java, Indonesia, 1987-1992. Part 2: Malignant tumours. AB - A total of 2193 tumours of the mouth and jaw diagnosed at the Laboratorium Patologi Anatomi Fakultas Kedokteran Universitas Airlangga, Indonesia from 1987 to 1992, inclusive, was studied. Malignant tumours constituted 45.3% of the lesions. Almost 71% of the malignant tumours were squamous cell carcinomas. The remainder were salivary gland tumours (21.5%) and sarcomas (4.5%). The male to female ratio for malignant tumours was 5.1:4.7. The incidence of malignant tumours per 100,000 population over the 6-year study period was 2.64. The yearly incidence seemed to increase except in 1990, when it dropped. The incidence of squamous cell carcinoma over the 6 years was 2.1. Calculation of the odds ratio suggested that people aged 40 and over are 5.8 times more likely to develop squamous cell carcinoma. PMID- 11735144 TI - International exchange in oral and maxillofacial surgery: comparison of the training pathways in the UK and USA at the end of the twentieth century. AB - The training of oral and maxillofacial surgeons varies considerably throughout the world and is reflected in the length of programme and scope of practice. The duration of training and workload for British and American trainees have long been topics of discussion and debate. The purpose of this paper was to compare the general training pathways of oral and maxillofacial surgeons in the UK and USA, as exemplified by the programme at the University of California in San Francisco, and in the UK by that at the East Lancashire Oral and Maxillofacial Unit. PMID- 11735145 TI - Oncofetal fibronectin and oral squamous cell carcinoma. AB - Fibronectin is a cell matrix glycoprotein, which exists as a number of isoforms that are often found within the cell matrix that surrounds tumours. Collectively these tumour-associated isomers of fibronectin have been termed oncofetal fibronectin (OFFN). We looked for expression of OFFN within oral squamous cell carcinomas (SCC) and related its presence to prognosis. The investigation used a monoclonal antibody (MoAb 5C10) to the glycosylated variant of OFFN, and 100 archival specimens of oral SSC. Immunostaining for OFFN was intense in the adjacent stroma of 43 squamous carcinomas, weak in 27 and absent in 30. Cervical metastases were found in 17/27 (63%) specimens that stained intensely, 6/17 (35%) that stained weakly and 3/13 (23%) that did not stain. Of the 21 cases which had extracapsular lymph node spread, 81% were from those that stained intensely, 19% from those that stained weakly and none from those that did not stain for OFFN expression. Also, 21/44 patients (49%) died in group with intense OFFN staining, 6/26 (23%) in the group with weak staining and 3/30 (10%) in the group that did not stain. The presence of OFFN glycoprotein in oral SCC as evaluated by immunostaining with MoAb 5C10 correlates strongly with the presence of metastatic lymph node involvement, particularly extracapsular involvement, and mortality. We therefore suggest that the degree of expression of OFFN in tumours is a valuable prognostic indicator. PMID- 11735146 TI - Glomus tumour of the hard palate: case report and review. AB - Glomus tumours are usually small benign neoplasms that occur in the dermis or subcutaneous tissues of the extremities. They are rare in the oral cavity, only 18 cases having been reported in the medical journals to our knowledge. They arise from the epithelioid glomus cells that normally reside in the stratum retinaculare corii of the skin and are concerned with temperature regulation. We report the clinical, morphological and immunohistochemical features of a case of glomus tumour of the hard palate. PMID- 11735147 TI - Congenital atresia of the orifice of the submandibular duct: a report of 2 cases and review. AB - Two infants presented with unilateral cystic swellings in the floor of the mouth as a result of imperforate submandibular ducts. This is thought to result from a congenital failure of canalization of the terminal end of the duct. Both cases responded to simple incision and decompression of the fluid-filled duct. Early treatment is important to avoid feeding difficulties and to prevent later complications such as ranula or sialadenitis. PMID- 11735148 TI - Re: Paoli J F et al. Craniofacial differences according to the body mass index of patients with obstructive sleep apnoea syndrome: cephalometric study in 85 patients. Br J Oral MaxilloFac Surg 2001; 39: 40-45. PMID- 11735149 TI - Re: Isaac AM, Moss CE, Whear NM. Unilateral temporalis muscle hypertrophy managed with botulinum toxin type A. Br J Oral Maxillofac Surg 2000; 38: 571-572. PMID- 11735150 TI - Re: Care in the placement of bicortical intermaxillary fixation screws. PMID- 11735151 TI - Re: Wake MJC. Descriptive titles: primary and secondary cleft surgeons. Br J Oral Maxillofac Surg 2001; 39:160. PMID- 11735152 TI - Re: Holmes S, Hutchison I. Reconstruction of the orbital floor after its removal for malignancy. Br J Oral Maxillofac Surg 2001; 39: 158-159. PMID- 11735153 TI - Re: Holmes S, Ali N, Bradley PF. A true craniomaxillary fracture. Br J Oral Maxillofac Surg 2001; 39: 160-161. PMID- 11735154 TI - Re: Iidas et al. Plunging ranula as a complication of intraoral removal of a submandibular sialolith. Br J Oral Maxillofac Surg 2001; 39: 214-216. PMID- 11735155 TI - Re: Whear NM, and Lopes V. The great auricular nerve and the facial nerve. Br J Oral Maxillofac Surg 2001; 39: 162-163. PMID- 11735156 TI - Enophthalmos caused by metastatic breast carcinoma. PMID- 11735157 TI - Re: Anderson G, Lewthwaite A, Yeats N. quality of life following surgery for oral and pharyngealmalignancy. Br J Oral maxillofac surg 2001;39: 163. PMID- 11735159 TI - Haploidentical stem cell transplantation in leukemia. AB - In acute leukemia patients, infusing a megadose of extensively T-cell-depleted hematopoietic stem cells after an immuno-myeloablative conditioning regimen ensures sustained engraftment of full-haplotype mismatched transplants without graft-vs-host disease. Besides the conditioning regimen and the megadose of stem cells donor natural killer cell alloreactivity also plays a role in facilitating engraftment and in preventing relapse. Since our first successful pilot study, our efforts have concentrated on developing new conditioning regimens, optimizing the graft processing and improving the post-transplant immunological recovery. The results we have so far achieved in 112 very high-risk acute leukemia patients show that haploidentical transplantation is now a clinical reality. Because virtually all patients in need of a hematopoietic stem cell transplant have a full-haplotype mismatched donor, who is immediately available, a T-cell depleted mismatched transplant should be offered, not as a last resort, but as a viable option to high risk acute leukemia patients who do not have, or cannot find, a matched donor. PMID- 11735160 TI - Current opinion in essential thrombocythemia: pathogenesis, diagnosis, and management. AB - A working diagnosis of essential thrombocythemia (ET) is made in the presence of nonreactive thrombocytosis and after the exclusion of another chronic myeloid disorder that may mimic ET in its presentation. Clinically, ET is characterized by vasomotor symptoms, thrombohemorrhagic complications, recurrent fetal loss, and transformation of the disease into either myelofibrosis with myeloid metaplasia or acute myeloid leukemia. Median survival in the majority of patients is close to that of an age-adjusted normal population, and current therapy has not been shown to either retard or hasten leukemic transformation, which is reported to occur in 1% to 20% of patients. The use of hydroxyurea in high-risk patients with ET has reduced the incidence of thrombosis, and recent studies have suggested the value of keeping the platelet count below 400 x 10(9)/L in such cases. The incidence of thrombosis in low-risk patients may not be high enough to warrant the use of cytoreductive therapy. Although effective in controlling vasomotor symptoms, aspirin therapy has not been shown to influence the risk of either recurrent thrombosis or first-trimester miscarriage in ET. Recent laboratory studies have suggested that hematopoiesis in ET may not always be clonal. Similarly, there is substantial heterogeneity in both megakaryocyte/platelet surface expression of the thrombopoietin receptor (c-Mpl) and bone marrow microvessel density. Clinicopathologic correlates to these biologic parameters are currently being defined. PMID- 11735161 TI - Arsenic trioxide: its role in acute promyelocytic leukemia and potential in other hematologic malignancies. PMID- 11735162 TI - Thrombolytic therapy: 2001. PMID- 11735163 TI - Desmoid tumours. AB - Desmoid tumours exhibit fibroblastic proliferation and arise from fascial or musculoaponeurotic structures. Despite their benign microscopic appearance, and their negligible metastatic potential, the propensity of desmoid tumours for local infiltration is potentially significant in terms of deformity, morbidity and mortality due to pressure effects and obstruction of vital structures and organs. The rarity of desmoid tumours, coupled with the variability in their clinical course, renders these lesions a vexing entity, and makes demonstration of the efficacy of any specific intervention difficult. Failure to recognize the potential for malignant behaviour in this tumour renders desmoids susceptible to inadequate treatment. This distinct pathological entity is reviewed with a specific focus on aetiology and treatment. PMID- 11735164 TI - The significance of intramammary nodes in primary breast cancer. AB - The use of sentinel node biopsy in primary breast cancer raises many new controversies with regard to extra-axillary nodes. Three cases with intramammary nodes are discussed in relation to sentinel node biopsy. PMID- 11735165 TI - ASCOT: a comprehensive clinical database for gastro-oesophageal cancer surgery. AB - We report the development of a multicentre prospective database for gastro oesophageal cancer surgery. The ASCOT project aims to collect detailed prospective data on co-morbidity, surgery, stage and outcome in a large cohort of patients from a wide variety of hospitals in the UK. We describe the rationale for the initiative, the process of defining the dataset, the database software and structure, the system for data recording and retrieval and the operating rules of the ASCOT co-operative group. Thirty Trusts are currently submitting data, and the first 1000 cases have now been entered. A first annual report has been produced, showing anonymized comparative figures for patient characteristics and outcomes of interest, and a second is due shortly. The collection of detailed comparable data, including co-morbidity evaluation, on a large scale is likely to prove valuable for unit specialist accreditation, surgeon re-validation, audit and research. PMID- 11735166 TI - Tamoxifen inhibits endothelial cell proliferation and attenuates VEGF-mediated angiogenesis and migration in vivo. AB - INTRODUCTION: Angiogenesis is fundamental to tumour growth and vascular endothelial growth factor (VEGF) is one of the most potent proangiogenic cytokines known. We have previously demonstrated that tamoxifen reduces serum VEGF in certain cancer patients. We hypothesized that tamoxifen may attenuate the angiogenetic response to VEGF. METHODS: Human dermal microvessel endothelial primary cell cultures (HMEC) were incubated with tamoxifen (1.25-5.0 microg) or vehicle. Cell proliferation was quantified using 5-bromo-2'-deoxyuridine (BrdU) labelling endothelial cell proliferation assay. The effect of oral tamoxifen (20 mg/day) on VEGF-mediated angiogenesis in vivo was assessed using a Matrigel angiogenesis assay in the Sprague-Dawley rat. RESULTS: Tamoxifen (5.0 microg/ml) significantly reduced HMEC proliferation over 24 h when compared with cells treated with vehicle alone. Oral administration of tamoxifen in the rat (20 mg/day) significantly reduced endothelial cell proliferation and migration in response to VEGF. CONCLUSION: Tamoxifen (5.0 microg/ml) reduces proliferation of a VEGF-dependent endothelial cell line in vitro. In vivo, orally administered tamoxifen reduces VEGF-mediated angiogenesis in the rat. These findings indicate that tamoxifen may directly inhibit the effect of VEGF on the endothelial cell, in addition to its previously described effect of reducing serum VEGF levels. This data supports a role for tamoxifen in modulation of the VEGF-dependent angiogenic response to surgical trauma, particularly as an adjuvant therapy for VEGF-dependent tumours. PMID- 11735167 TI - Extranodal extension of axillary metastasis of invasive breast carcinoma as a possible predictor for the total number of positive lymph nodes. AB - INTRODUCTION: By the implementation of the sentinel node procedure in the treatment of breast carcinoma routine axillary lymph node, dissection can be abandoned in patients with a tumour-negative sentinel node. When the sentinel node is positive there are two options; an axillary dissection or radiotherapy of the axilla. In the latter case one is not informed about the total number of positive lymph nodes which can be of importance for the choice of adjuvant chemotherapy. In this paper we analyse whether it is possible to use histological parameters of a lymph-node metastasis to predict the number of tumour-cell containing nodes. METHODS: Four hundred and ninety-eight patients treated for invasive breast cancer at our department from 1991 to 1996 were investigated to see whether extranodal extension of axillary metastases had a significant predictive value for the number of positive lymph nodes. Extranodal extension was scored as: no extranodal extension (NEE) and extranodal extension (EE); the latter was subdivided in minimal extranodal extension (MEE) or extensive extranodal extension (EEE). RESULTS: Of 498 patients, 230 patients had axillary involvement. NEE was seen in 83 (36.1%) patients and 147 (63.9%) had EE. Subdivision of this latter group revealed 77 patients with MEE (52%), 65 patients with EEE (45%) and five patients not further specified (3%). The number of positive nodes for the EE-group (6.9+/-0.5) was significant higher compared with the NEE-group (2.1+/-0.2) (P<0.001). The number of positive nodes was also significantly higher for the EEE-group compared to the MEE-group, 10.6+/-0.8 vs 4.0+/-0.4 (P<0.001). The predictive value for > or =4 positive axillary lymph nodes was 84.6% for EEE, 58.5% for EE and only 14.5% for NEE. These differences were significant (P<0.001). CONCLUSIONS: The presence of extranodal extension in axillary lymph-node metastases can be a good predictor for the expected number of positive nodes. This histological parameter could be of value to determine the kind of adjuvant chemotherapy after a positive sentinel-node biopsy with only radiotherapy of the axilla and no further axillary lymph-node dissection. PMID- 11735168 TI - Outcome and predictive factors of local recurrence and distant metastases following primary surgical treatment of high-grade malignant phyllodes tumours of the breast. AB - AIM: High-grade malignant phyllodes tumour (MPT) is a rare but aggressive breast malignancy and forms approximately 25% of all phyllodes tumours. The aim of the study was to determine parameters that influence outcome in high-grade MPT. METHODS: This study consisted of 48 women diagnosed with high-grade MPT. All patients were treated primarily with surgery by local excision (LE, margins <1 cm), wide local excision (WLE, margins > or =1 cm) or mastectomy. Cox's regression was used for multivariate analysis of the data. RESULTS: The mean patient age was 47 (range 21-85) years and the average tumour size was 7.8 (range 1.5-20) cm. Ten patients were treated with LE, 14 with WLE and 24 with mastectomy. The median follow-up was 9 years (range 5 months-28 years). Local recurrence (LR) occurred in 19 patients (40%) at mean time of 28 (range 5-84) months after primary treatment. Distant metastasis (DM) occurred in 13 (27%) patients at average time of 25.6 (range 6-120) months. LR, subsequent metastatic spread and survival following treatment of MPT were related to tumour size and excision margins, but not to other clinical or histopathological characteristics. CONCLUSION: Tumour size and surgical margins were found to be the principal determinants of LR and DM. Complete surgical excision, by mastectomy if necessary, is important in the primary surgical treatment of high-grade MPT. PMID- 11735169 TI - Pre-operative serum levels of sialyl Tn antigen predict liver metastasis and poor prognosis in patients with gastric cancer. AB - AIMS: To clarify the prognostic value of preoperative serum levels of sialyl Tn antigen (STN) for survival of gastric cancer patients. METHODS: Pre-operative serum levels of STN, sialyl Lewis(a)antigen (CA19-9) and carcinoembryonic antigen (CEA) were examined in 180 patients who underwent resection of gastric cancer. Patients were divided into high and low antigen groups on the basis of a selected diagnostic-based cut-off value. Correlation between high antigen serum levels, established clinicopathologic factors and prognosis was examined by univariate and multivariate analysis. RESULTS: Twenty-eight patients (15.6%) were classified as high STN; 37 (20.6%) as high CA19-9; and 33 (18.3%) as high CEA. The survival time of the high STN, CA19-9 or CEA group was shorter than that of the respective low-antigen group (P<0.0001, P=0.0008 or P=0.0002, respectively). Patients with stage III/IV tumours with high STN had a shorter survival time that those with low STN (P=0.0004). Cox's regression with multiple covariates showed that high serum STN is an independent factor predicting a worse outcome in gastric cancer patients. Multiple logistic regression analysis revealed that high serum STN is an independent predictor for the development of liver metastasis. CONCLUSIONS: Pre-operative high serum levels of STN predict both liver metastasis and poor prognosis after resection for gastric cancer. PMID- 11735171 TI - Coeliac plexus block: utility of the anterior approach and the real time colour ultrasound guidance in cancer patient. AB - AIMS: The aim of our study was to demonstrate the efficacy of ultrasound-guidance compared with computed tomography (CT) guidance for coeliac plexus block in cancer patients. METHODS: Coeliac plexus block (30 ml ethanol) was performed in 34 cancer patients (sex ratio: 10F, 24M), mean age: 54.8 years (range 26-67) under CT (n=21) and ultrasound-guidance (n=13). All patients had excruciating epigastric and generalized abdominal pain caused by cancer of the pancreas (n=13) or upper abdominal viscera (n=9) or a malignancy of extra-digestive origin (n=12). Feasibility and complication rates were analysed. RESULTS: Notable pain relief was obtained in 27 (79%) of the patients. The technical success rate was 100% for CT-guidance and 93% (13/14) for ultrasound guidance. There were six minor complications (17%): chemical peritonitis (n=2), orthostatic arterial hypotension (n=2) and transient left shoulder pain (n=2), no major complications occurred. The target route was transhepatic in 6/13 of the ultrasound cases and mean length was 6 cm (range 3-12 cm). Colour Doppler sonography improved visualization of the 21 Gauge Chiba needle when the needle shaft was vibrated. Echogenic foci were observed around the origin of the coeliac trunk and superior mesenteric artery in all cases. CT coeliac block was successfully performed after failure of ultrasound guidance in one patient. CONCLUSION: Ultrasound guidance is safe and effective and should be attempted for coeliac plexus block whenever possible. PMID- 11735170 TI - Micrometastases in bone marrow of patients with suspected pancreatic and ampullary cancer. AB - AIMS: This prospective study aimed to evaluate the detection of micrometastases in bone marrow of patients with suspected pancreatic and ampullary cancer and to determine their predictive value on overall survival. METHODS: Between December 1997 and December 1998, 35 patients (19 male, 42-77 years) with suspected pancreatic and ampullary cancer underwent diagnostic laparoscopy as a final staging procedure before exploration. Bone marrow was aspirated from the iliac crest at the beginning of laparoscopy. Mononuclear cells were isolated and stained using the specific monoclonal antibody CAM 5.2. RESULTS: Cytokeratin positive cells were detected in 12/35 (34%) of all patients. In the 31 patients with a final diagnosis of carcinoma, a positive staining was found in 10/31 (32%) of the bone marrow aspirates. After a median follow-up of 17 months (2-24), 15/31 (48%) patients had died: 7/10 (70%) with and 8/21 (38%) without micrometastases (* P<0.04). All four patients who turned out to have chronic pancreatitis were alive without malignancy. In two of these four patients, distinct cytokeratin positive cells were seen. CONCLUSIONS: Micrometastases in bone marrow of patients with the final diagnosis pancreatic or ampullary carcinoma seem to predict a significantly shorter survival. However, clinical use of cytokeratin markers cannot be recommended at present, because false-positive staining was found. PMID- 11735172 TI - Prospective randomized study of post-operative chemotherapy with levamisole and UFT for head and neck carcinoma. AB - AIMS: A prospective randomized study was conducted to evaluate the benefit of adjuvant levamisole/UFT (futraful and uracil) chemotherapy in head and neck squamous cell carcinoma. METHODS: Sixty-five patients with stage III and IV squamous cell carcinomas of oral cavity, oropharynx, hypopharynx and larynx with no distant metastasis were randomized for the chemotherapy study. Thirty-one patients were randomized for chemotherapy and two of them were subsequently excluded. In this study, a total of 29 patients on levamisole/UFT therapy and 34 patients on the control group were analysed. The main outcome was measured by the 5-year disease-free actuarial survival rate. RESULTS: The rates of distant metastasis were 10% for chemotherapy group and 32% for control group (P=0.06). The 5-year disease-free actuarial survival rates for patients with and without adjuvant chemotherapy were 57% and 39% respectively (P=0.207). CONCLUSIONS: A trend of better distant control in head and neck cancer patients with post operative adjuvant oral chemotherapy was observed. The side effects were minimal. However, there was no statistically significant improvement in the overall long term survival. It may be of value to conduct a large-scale multi-centre prospective randomized study to verify the efficacy of levamisole and UFT as post operative adjuvant chemotherapy for the control of distant metastasis in high risk population. PMID- 11735174 TI - Apoptosis in vascular disease. PMID- 11735173 TI - The significance of human telomerase reverse transcriptase (hTERT) in cancer. AB - Since the discovery of the enzyme telomerase in humans, it has become apparent that it is the most general of tumour markers known, and enormously significant in its potential for diagnostic, prognostic and therapeutic applications. Extensive work has identified three core components of the enzyme, of which the catalytic subunit hTERT (human telomerase reverse transcriptase) appears to be the most important. The aim of this article is to review the current evidence for the function and activity of hTERT in malignant conditions, and to discuss the future possibilities in terms of cancer diagnosis and treatment. PMID- 11735175 TI - Peripheral vascular surgery and magnetic resonance arteriography--a review. AB - OBJECTIVES: to review the current status of lower limb MRA. DESIGN: a literature review based predominantly on a MEDLINE database search of English-language publications from January 1991 to October 2000. MATERIALS AND METHODS: twenty eight articles, concerning non-enhanced MRA (13), gadolinium-enhanced MRA (14) or both (1), met the predefined requirement for quality. Results gadolinium-enhanced MRA (CE-MRA) seems to be more accurate, quicker and associated with fewer problems than non-enhanced (TOF) MRA. TOF-MRA has a sensitivity and specificity of 93% (range 64-100%) and 88% (range 57-100%) respectively, and CE-MRA presents values of 96% (range 71-100%) and 96% (63-100%), respectively, using conventional arteriography as the gold standard. Some articles report a substantial incidence of runoff vessels suitable for distal bypass visible on MRA but invisible on conventional arteriography. Gadolinium contrast is given intravenously and is generally well tolerated and has no known nephrotoxicity. CONCLUSION: CE-MRA is accurate compared to conventional arteriography, has the potential to increase the limb salvage rate for selected patients, is non-invasive and well tolerated. PMID- 11735177 TI - Remifentanil conscious sedation during regional anaesthesia for carotid endarterectomy: rationale and safety. AB - OBJECTIVES: to prospectively evaluate the safety and efficacy of remifentanil during regional anaesthesia for carotid endarterectomy. METHODS: twenty-eight consecutive patients underwent carotid endarterectomy with combined superficial and deep cervical plexus block supplemented with continuous intravenous 0.04 microg.kg(-1).min(-1)remifentanil infusion. Depth of sedation was monitored using the Observer's Assessment of Alertness/Sedation Scale (OAA/S). The degree of pain, discomfort and anxiety was self-assessed by the patients using a horizontal visual analogue scale. RESULTS: all patients experienced adequate comfort and analgesia. No local anaesthetic supplementation was necessary. No patient had a OAA/S score lower than 4 (with 5=awake/alert to 1=asleep). Respiratory depression did not occur. Selective shunting was required in four cases. No patient was converted to general anaesthesia. There were no permanent neurological deficits, cardiopulmonary complications or deaths. CONCLUSION: remifentanil as a supplement to regional anaesthesia for carotid endarterectomy, provides comfort and analgesia without hampering mental status evaluation. PMID- 11735179 TI - Use of the percutaneous vascular surgery device for closure of femoral access sites during endovascular aneurysm repair: lessons from our experience. AB - OBJECTIVES: to evaluate the results of our early experience with a percutaneous closure device for aortic aneurysm repair and to identify device related and patient related factors leading to procedure failure. METHODS: eighty-two percutaneous closures in forty-four patients was performed using the 10F Prostar XL Percutaneous Vascular Surgery device during the repair of 1 iliac, 1 thoracic and 42 abdominal aortic aneurysms. RESULTS: successful closure was achieved in 70 access sites (85%) with 12 sites requiring conversion to an open groin incision. The reasons for failure include difficult device introduction due to a tortuous iliac, deflection of needles due to previous scar, femoral artery occlusion and failure of the device to close the arteriotomy. There was one intraoperative death from retroperitoneal haemorrhage and another patient developed a pseudoaneurysm at the cannulation site. CONCLUSIONS: use of the percutaneous closure device requires very careful patient selection. Preoperative radiological assessment of the ilio-femoral vessels is vital to assess for cacification and tortuosity. High device failure rates can be expected from obese patients and those with scarred groins. When difficulty is encountered during the procedure, there should be a low threshold for conversion to an open groin incision. The device and the method of introduction can be further improved to address some of these issues. PMID- 11735178 TI - Spinal cord metabolism during thoracic aortic cross-clamping in pigs with special reference to the effect of allopurinol. AB - OBJECTIVE: investigate the metabolic response of the spinal cord and the effect of allopurinol following cross clamping of the descending thoracic aorta in a porcine model. DESIGN: experimental animal study. MATERIALS: twelve domestic swine. Six pigs were pre-treated with allopurinol, while six pigs served as controls. METHODS: measurement of extracellular concentrations of glucose, pyruvate, lactate, glycerol and glutamate using microdialysis in the lumbar spinal cord. Measurement of lumbar spinal blood flow using laser Doppler technique. RESULTS: for all animals there was a significant decrease in concentrations of glucose and pyruvate together with a significant increase in the lactate-pyruvate ratio during aortic cross clamping. There was also a significant increase in glycerol concentrations 60 min after cross clamping, and a significant decrease in glutamate concentrations after 50 min. No differences in concentrations of glucose, pyruvate, lactate and glutamate or the glutamate pyruvate ratio were observed between animals used as controls and those treated with allopurinol. The laser Doppler flux decreased to 40% of pre cross-clamp level, returning to normal values at declamping. CONCLUSION: the changes in energy-related metabolites reflect a considerable ischaemia in the spinal cord tissue but there was no convincing effect of allopurinol on the lumbar spinal cord metabolism during thoracic aortic cross clamping in this model. PMID- 11735180 TI - The value of the peak systolic velocity ratio in the assessment of the haemodynamic significance of subcritical iliac artery stenoses. AB - OBJECTIVES: iliac stenoses with a PSV ratio between 2.0 and 3.0 in patients with symptomatic arterial obstructive disease of the legs might be misinterpreted when compared with intra-arterial pressure measurements (IAPM). The aim of this study was to compare the value of the PSV ratio with IAPM as the reference standard in the assessment of the haemodynamic significance of subcritical iliac artery stenoses (iliac stenosis with PSV ratio between 1.5 and 3.5). DESIGN, PATIENTS AND METHODS: fifty-eight iliac tracts in 53 consecutive patients with symptomatic arterial obstructive disease of the legs with an isolated stenosis with PSV ratio between 1.5 and 3.5 were studied prospectively. The results of those iliac duplex scanning were compared to IAPM. Results a poor agreement was found between IAPM and PSV ratios. For the PSV ratios > or =2.0, 2.5 and 3.0 the sensitivities were 74%, 37% and 16%, respectively. The specificities were 70%, 90% and 95%, the positive predictive values 82%, 88% and 86%, respectively, and the negative predictive values 58%, 43% and 37%, respectively. CONCLUSION: the results of this study show that the PSV ratio parameter is not accurate enough to evaluate the haemodynamic significance of subcritical iliac artery stenoses. PMID- 11735181 TI - Mortality and morbidity following endovascular repair of abdominal aortic aneurysms: analysis of two single centre experiences. AB - OBJECTIVE: to show how differences in anatomical and physiological risk factors can affect the outcome of endovascular repair of AAA by describing the experience of two centres with different selection policies. METHODS: one hundred and thirty five patients (group I) were treated at Queen's Medical Centre (Nottingham, U.K.) using 101 in-house made and 34 manufactured stent-grafts. Median diameter, length and angulation of the proximal aneurysm neck were 26 mm, 27 mm, 40 degrees, respectively. Seventy-six patients had ischaemic heart disease, 47 had left ventricular failure, median forced expiratory volume in one second (FEV1) was 83%, median creatinine was 100 micromol/l and median age was 72 years. Fifty patients (group II) were treated at Timone Hospital (Marseilles, France) using seven in-house made and 43 manufactured stent-grafts. Median diameter, length and angulation of the proximal aneurysm neck were 25 mm, 34 mm, 33 degrees, respectively. Thirteen patients had ischaemic heart disease, two had left ventricular failure, median forced expiratory volume in one second was 101%, median creatinine was 108 micromol/l and mean age was 72 years. RESULTS: anatomical characteristics of the proximal neck were significantly worse in group I (p=0.02 for the three variables). Cardiac comorbidities were more frequent and mean FEV1 was lower in group I (p<0.0001 and p=0.001, respectively. Median aneurysm diameter was significantly greater in group I (65 mm) than in group II (53 mm) (p<0.001). Postoperative mortality was 9% and 0% in groups I and II respectively (p=0.03). The incidence of technical complications (groin wound complications and side branches endoleaks being excluded) was 20% and 0% in groups I and II, respectively (p=0.0006). CONCLUSION: postoperative mortality and technical complication rates were significantly greater in group I than in group II, readily explained by poorer general condition and worse anatomical characteristics of the proximal neck in group I. PMID- 11735182 TI - The evolving technique of endovascular stenting of abdominal aortic aneurysm; time for reappraisal. AB - PURPOSE: to investigate the occurrence of intra-operative device-related complications during stent-grafting for abdominal aortic aneurysm. MATERIALS AND METHODS: data on patient characteristics, vascular morphology, operative technical details, procedural and device-related complications were obtained from the European collaborators on stent-graft techniques for abdominal aortic aneurysm repair (EUROSTAR) registry. Only intra-operative device-related complications were taken into account. Potential risk factors for device-related complications were examined by logistic regression analysis. The association between these complications and conversion to open surgery and death were determined. RESULTS: between January 1994 and July 2000, 2862 patients in 90 participating hospitals underwent endovascular abdominal aortic aneurysm repair. Device-related complications occurred in 238 (8.3%) patients. Complications were associated with the age of the patient (p=0.002), gender (p=0.05), smoking habit (p=0.001), pre-operative aneurysm diameter (p=0.005), type of device implanted (p=0.0001), fitness of the patient for open surgery (p=0.002), and year of operation (p=0.001). Adjusted for risk factors, the occurrence of complications decreased between 1994 to 2000 from 21.7% to 7.3%, respectively. Patients with device-related complications were 13.6 times (95% CI; 9.2-20.1) more likely to have conversion to an open procedure and 2.4 times (95% CI; 1.4-4.0) more likely to die within 30 days of the operation. CONCLUSIONS: intra-operative device related complications were common, although appear to be decreasing in frequency, and were significantly related to conversion and post-operative death. PMID- 11735183 TI - In vitro degradation of aortic elastin by Chlamydia pneumoniae. AB - OBJECTIVES: to investigate whether Chlamydia pneumoniae (C. pneumoniae) may increase elastin degradation in the aortic wall. MATERIALS AND METHODS: eighteen full thickness aortic wall samples from non-aneurysmal infrarenal abdominal aortas were collected from autopsies. Two adjacent and equally large pieces were cut out of each aortic sample. From each sample, one piece was incubated in a HEp 2 cell culture infected with C. pneumoniae and the other piece was incubated in an uninfected HEp-2 cell culture. The incubation time was one week at 35 degrees C. The concentration of elastin-derived peptides (EDP) (ng/ml) in the medium of each cell culture was measured in duplicate. For each paired sample, delta-EDP (EDP in HEp-2 cell culture infected with C. pneumoniae- EDP in uninfected HEp-2 cell culture) was calculated. RESULT: there was a significantly increased degradation of aortic elastin, estimated by EDP concentrations in cell culture conditioned medium, when aortic wall samples were incubated in C. pneumoniae cultures compared with uninfected cultures (p=0.025, Wilcoxon signed ranks test). CONCLUSION: these results indicate that there is a relationship between the presence of C. pneumoniae and increased elastin degradation in the aortic wall in vitro. This suggests C. pneumoniae in the aortic wall directly or indirectly leads to the degradation of aortic elastin. PMID- 11735184 TI - A 16-year haemodynamic follow-up of women with pregnancy-related medically treated iliofemoral deep venous thrombosis. AB - OBJECTIVES: to evaluate clinical and functional long-term outcomes following pregnancy-related medically treated iliofemoral deep venous thrombosis (DVT). DESIGN: retrospective follow-up of patients identified through a registry search. MATERIAL AND METHODS: twenty-five women underwent clinical examination, colour duplex ultrasound and computerised strain-gauge plethysmography on two occasions a mean of nine and 16 years after DVT. RESULTS: 40% of the patients were completely asymptomatic and 52% had no clinical signs of venous disease after a mean follow-up of 16 years. The clinical signs were in general mild, and none of the 25 patients had skin changes or ulcers. Deep venous reflux was found in 36% of the patients; the same percentage at nine- and 16-years follow-up, and 24% had normal ultrasonographic appearance of all deep veins. None of the patients had plethysmographic evidence of outflow obstruction. There was a significant relationship between measures of venous reflux and the presence of leg swelling, but there was no clear relation between functional abnormalities and the extent of the initial DVT. CONCLUSION: even after 16 years there are relatively mild symptoms and signs of venous disease in women with medically treated pregnancy related iliofemoral DVT. Our results do not support earlier stated opinions that these patients represent a particular risk group for developing post-thrombotic syndrome. PMID- 11735185 TI - "Long haul" flight and deep vein thrombosis: a model to help investigate the benefit of aspirin and below-knee compression stockings. AB - OBJECTIVE: to develop a model simulating factors of "long haul" flight to investigate the relationship with DVT. MATERIALS AND METHODS: volunteers (19 males: 20 females) sat for 6 h in a warm (>25 degrees C), dry environment, limited in movement, consuming alcohol (40 ml of 40% alcohol/hour) and salted foods (300 g). Half of the subjects received 150 mg aspirin and wore especially designed below-knee, compression stockings (Class 1 profile). Changes in full blood counts were recorded, and as an indication of DVT formation, plasma was analysed for D-dimer. Limb swelling was assessed from leg measurements. RESULTS: after 6 h, in controls, there were significant rises in platelet packing (Pct p<0.04), total platelet numbers (p<0.003) and total numbers of white blood cells (WBC's p<0.001). With aspirin plus stockings, there were similar significant rises in total platelet numbers (p<0.002) and total WBC's (p<0.001). In both groups, significant rises were seen in all WBC types (except basophils). Wearing compression stockings prevented calf swelling seen in controls after 6 h (p<0.002). No subject developed a DVT, or a change in levels of D-dimer. CONCLUSION: changes in the cellular components of blood, particularly WBC's, combined with vaso-compression and reduced flow could predispose towards DVT. Aspirin, combined with compression stockings, may provide prophylaxis. PMID- 11735186 TI - A ruptured abdominal aortic aneurysm repaired with a bifurcated unibody endoluminal graft. PMID- 11735187 TI - Outcome of iliac kissing stents. PMID- 11735188 TI - Perigraft seromas complicating prosthetic bridge arteriovenous fistula--solution with autogenous vein interposition. PMID- 11735189 TI - Acute forefoot ischaemia: an unreported complication of dorsalis pedis artery aneurysm. PMID- 11735190 TI - Interferon induced severe digital ischaemia in chronic myeloid leukaemia- successful treatment using ketanserin. PMID- 11735191 TI - Bilateral carotid endarterectomy. PMID- 11735192 TI - Patient scoring systems. PMID- 11735194 TI - Haemorrhage associated with combined clopidogrel and aspirin therapy. PMID- 11735196 TI - Cytokines and inflammatory pathways in the pathogenesis of multiple organ failure following abdominal aortic aneurysm repair. AB - Multiple organ failure is a common mode of death following abdominal aortic aneurysm repair, particularly after rupture. Cytokines are the principal mediators of the inflammatory response to injury and high levels of circulating cytokines have been associated with poor outcome in major trauma and sepsis. Abdominal aortic aneurysm repair results in an ischaemia-reperfusion injury to the tissues distal to the site of aortic clamping. The inflammatory response in these tissues causes the release of cytokines, principally Interleukins 1-beta, 6, and 8, and Tumour Necrosis Factor alpha. If released in large enough concentrations, these cytokines may enter the circulation and gain access to organs distant to the site of initial injury. Circulating cytokines cause dysfunction of the renal, cardiovascular, respiratory, nervous and musculo skeletal systems. The combination of these individual changes in organ function is the multiple-organ dysfunction syndrome, which may progress to multiple organ failure. PMID- 11735197 TI - Patients' thromboembolic potential between bilateral carotid endarterectomies remains stable over time. AB - OBJECTIVES: There is limited understanding of the reasons underlying post-CEA carotid thrombosis. Clinicians have often implicated operative technique, such as patch type or shunting, however the evidence for this is limited. We have studied whether it is the patients themselves who are prothrombotic, by studying the rates of emboli detection in patients undergoing bilateral CEAs at separate time points. MATERIALS AND METHODS: Sixteen patients (3 women) underwent CEA during the study period, all of whom were taking aspirin. CEA was performed in a standardised manner throughout the study. All patients were monitored for 3 h postoperatively using a 2 MHz fixed head probe. RESULTS: Those patients who had no emboli detected on TCD after the first operation, had a mean of 2.5 emboli after the second operation. Patients with emboli after the first operation had a mean of 41.3 emboli after the second CEA (MWU test, p=0.02). The dose of aspirin administered did not affect emboli rates. Correlation of the number of emboli detected after the first CEA with the second CEA gave a significant correlation ( p=0.038). CONCLUSIONS: There appear to be factors relating to the patient that places some individuals at an increased risk of thrombotic stroke. Further elucidation of these factors may enable more effective, targeted therapy to be applied in the prevention of arterial thrombosis. PMID- 11735198 TI - The role of myocardial perfusion scanning, heart rate variability and D-dimers in predicting the risk of perioperative cardiac complications after peripheral vascular surgery. AB - OBJECTIVES: To study the value of a number of proposed prognostic factors in prediction of the risk of perioperative cardiac events after vascular surgery. DESIGN AND METHODS: Two hundred and ninety-seven patients undergoing peripheral vascular surgery were prospectively studied. Patients underwent preoperative 24 h ambulatory electrocardiography, measurement of haemostatic variables, myocardial assessment of perfusion by dipyridamole-thallium scintigraphy and radionuclide ventriculography. The primary endpoint was cardiac death or nonfatal myocardial infarction within 30 days of surgery. A combined endpoint included the primary endpoint plus occurrence of cardiac failure, unstable angina or serious arrhythmias. RESULTS: The primary endpoint occurred in 21 (7%), and the combined endpoint in 41 (14%) of patients. On multivariate analysis, increased age, previous myocardial infarction, aortic surgery, impaired heart rate variability and a positive thallium scan were independent predictors of primary end-points. Preoperative atrial fibrillation and increased fibrin D-dimer were additional predictors of the combined endpoint. Construction of receiver-operator characteristic curves to examine the incremental value of predictive models showed that sensitivity and specificity of clinical data alone for primary endpoints was 71% and 72% respectively, while for the full model (incorporating heart rate variability and thallium data) this rose to 84% and 80% (p=0.0001). CONCLUSIONS: Preliminary screening using clinical data has limited value in risk assessment prior to vascular surgery but preoperative heart rate variability, D dimers and thallium scanning provide modest incremental predictive value. PMID- 11735199 TI - The use of impedance index in the surveillance of PTFE femorodistal grafts. AB - BACKGROUND: Impedance is the equivalent in pulsatile flow of resistance in steady flow. The impedance index has been used successfully in the surveillance of vein grafts, but its use has not been reported in the context of PTFE femorodistal grafts. METHODS: Twenty-eight patients (median age 68 years (IQR 59-73 years) and 20 men) undergoing 28 PTFE femorodistal grafts with a vein cuff were evaluated prospectively comparing the impedance index with standard duplex graft surveillance. All grafts were performed for critical ischaemia. At risk grafts were identified and treated appropriately after angiography. RESULTS: The primary patencies at 1 and 2 years were 82% and 50% respectively. Duplex identified 11 at risk grafts of which 9 had an identifiable correctable lesion. Impedance analysis overpredicted at risk status when compared with duplex in the immediate postoperative phase and was unsuccessful in detecting inflow disease or low flow relating to cardiac failure. Using a threshold index of 0.5, impedance analysis has a sensitivity of 87%, specificity of 88%, with positive and negative predictive values of 76% and 94% respectively. CONCLUSIONS: Impedance index is a non-invasive method of graft surveillance which is applicable to PTFE femorodistal bypasses and may be a useful alternative to duplex although formal validation studies will be required. PMID- 11735200 TI - A new device for the measurement of disease severity in patients with intermittent claudication. AB - OBJECTIVES: To assess a new method of determining functional impairment in patients with intermittent claudication, the Double Physiological Walking Test (DPWT) using the PADHOC (Peripheral Arterial Disease Holter Control) device, against a standard treadmill test. DESIGN: Patients with intermittent claudication presenting to the department were considered for both the DPWT and a standard treadmill test. METHODS: initial claudicating distance, maximal walking distance and speed of walking were determined for both parts of the DPWT. Initial claudicating distance and maximal walking distance were determined from the treadmill test. Comparisons were made between the treadmill test and the DPWT. RESULTS: The treadmill test was unable to be performed in 22% of patients due to defined contraindications. There were strong correlations in both walking distances and disease severity when comparing the DPWT and the treadmill test. Patients in whom the treadmill test was contraindicated had significantly shorter walking distances on the DPWT than those who were able to complete a treadmill walking test. CONCLUSIONS: The DPWT correlates strongly with walking distances obtained from a standard treadmill test. However, the PADHOC can be used in a number of differing locations and settings as well as in patients in whom a treadmill test is contraindicated. It therefore has a role to play in the initial assessment of patients presenting with intermittent claudication. PMID- 11735201 TI - Skeletal muscle ischaemia-reperfusion injury: further characterisation of a rodent model. AB - BACKGROUND: Postischaemic damage in skeletal muscle may be reflected in changes to microvascular blood flow, vascular permeability, and subsequent tissue viability. Previous preclinical studies have not addressed all these parameters, and have not used periods of ischaemia and reperfusion relevant to the clinical setting. This study aimed to develop an animal model hindlimb ischaemia reperfusion to simulate acute lower limb ischaemia. METHODS: A rodent model of hindlimb tourniquet-induced ischaemia-reperfusion was employed. Gastrocnemius muscle blood flow (GMBF; radio-labelled microspheres), oedema (GMO; using a wet:dry ratio method) and viability (GMV; histochemistry and computerised planimetry) were quantified. RESULTS: 6 h ischaemia per seresulted in neither muscle oedema nor loss of viability, but these changes were apparent following 4 h reperfusion. Early reperfusion at 10 min demonstrated low reflow, with GMBF improving at 120 min before declining sharply at 240 min. CONCLUSION: Prolonged hindlimb ischaemia followed by reperfusion in this rodent model caused significant reductions in gastrocnemius muscle blood flow, associated with muscle oedema and necrosis. These three parameters have not been previously reported together in the same model. This reproducible model could be used in the evaluation of potential therapeutic intervention strategies aimed at ameliorating skeletal muscle reperfusion injury. PMID- 11735202 TI - Endovascular repair of ruptured abdominal aortic aneurysm--a challenge to open repair? Results of a single centre experience in 20 patients. AB - INTRODUCTION: The mortality from ruptured abdominal aortic aneurysm (AAA) remains in the region of 50% despite advances in critical care. Endovascular repair of AAA has been shown to be associated with reduced physiological stress in the elective setting. It is hypothesised that the reduced physiological stress associated with EVAR may improve the outcome in patients with ruptured AAA. METHODS: A feasibility study of endovascular repair of ruptured AAA was undertaken at the University Hospital, Nottingham, U.K. between 1994 and 2000. Patients admitted with ruptured AAA were assessed by a team familiar with endovascular techniques for elective repair of AAA. After giving informed consent patients underwent spiral computed tomographic angiography (CTA) in the majority of cases. Patients were then transferred to the operating theatre for EVAR. RESULTS: Twenty patients underwent repair of ruptured AAA. Of these 20 patients, seven were referred from another hospital. Eight patients were considered unfit for open repair. The median duration of procedure was 180 min (range 120-480) and median blood loss was 1200 ml (range 750-2000 ml). The overall peri-operative mortality was 45%. A number of intra-operative and peri-operative procedures (both open and endovascular) were required to ensure aneurysm exclusion and to deal with the complications of endovascular surgery. CONCLUSIONS: Ruptured AAA remains a particularly hazardous condition to treat. There are a number of advantages of EVAR in this condition. A number of the problems early in the experience of EVAR have been addressed, but further experience is required to demonstrate its efficacy when compared with open repair. PMID- 11735203 TI - Perianeurysmal fibrosis: a relative contra-indication to endovascular repair. AB - OBJECTIVE: Perianeurysmal fibrosis (PAF) with involvement of neighbouring viscera can render open repair of inflammatory aneurysms technically difficult and therefore hazardous. For this reason, endovascular repair (EVAR) has been advocated as the preferred approach for this condition. EVAR is known to induce a systemic inflammatory response in patients but the nature of the local response remains unknown. If significant, such a response could exacerbate rather than ameliorate PAF. The aim of the study was to examine the incidence, course and consequences of perianeurysmal fibrosis detected by computerised tomography (CT) before and after EVAR. MATERIAL AND METHODS: The clinical records of patients treated by EVAR and followed for at least 6 months were reviewed. Pre and post operative CT images were independently graded for PAF by three radiologists according to a standard protocol. RESULTS: PAF was documented preoperatively in six out of a total of 61 patients. In two of these PAF worsened after EVAR resulting in ureteric obstruction and hydronephrosis requiring ureteric stents. In the remaining 4 patients PAF did not reduce postoperatively. PAF of low grade developed postoperatively in 10 out of 55 patients (18%) in whom there was no evidence of PAF on preoperative imaging. Median follow-up was 18 months (range 6 36 months). The development of periaortic fibrosis de novopostoperatively was statistically significant (McNemar's test p=0.002). CONCLUSION: EVAR does not seem to reverse PAF if this is present preoperatively and it induces this condition in approximately one sixth of patients without evidence of preoperative PAF. The potential for this adverse inflammatory local response should be taken into account when considering EVAR for treatment of aneurysms with perianeurysmal fibrosis and must be weighed against the perceived benefits of this approach. PMID- 11735204 TI - Ultrasound guided human thrombin injection. A new modality in the management of femoral artery pseudo-aneurysms. AB - OBJECTIVES: We report our initial results of a prospective study of duplex ultrasound-guided injection (UGTI ) of thrombin in the management of femoral artery pseudo-aneurysms. We used human thrombin to avoid the increase in the human antibodies directed against fibrinogen, with the use of bovine thrombus, that preclude further utilisation of the bovine fibrin glue during cardio thoracic surgery. METHODS: From 1999 to 2001, 19 patients, aged 69 (range 52-85) years presented with 21 femoral pseudo-aneurysms were treated. The mean pseudoaneurysm diameter was 30 (15-55) mm. All but two were secondary to cardiac procedures and the common femoral artery was the injured vessel in all instances. Patients were referred within 2-21 days following their iatrogenic injury. RESULTS: Immediate thrombosis of the sac occurred in 19 (90%) of the 21 pseudo aneurysms. After a second injection, complete occlusion occurred in the remaining two patients. Two patients (CI 95%; 1-19) with three femoral pseudo-aneurysms developed leg pain. Duplex ultrasound follow-up showed two recurrences (9.5% - CI 95%; 1-19) and both were treated by repeat UGTI. There was no conversion to surgical repair. CONCLUSION: This percutaneous minimally invasive technique is safe and effective in the management of iatrogenic pseudo-aneurysms in this high risk group of patients. Human thrombin has significant advantages over bovine thrombin. PMID- 11735205 TI - Serum elastin peptides in the preoperative evaluation of abdominal aortic aneurysms. AB - OBJECTIVE: Serum elastin peptides (SEP) have been reported to be associated with the expansion of small abdominal aortic aneurysms (AAA). Consequently, SEP measurements may predict future rupture, and allow further selection for surgery in cases referred for surgery due to size. MATERIAL AND METHODS: SEP was measured in 90 men and 10 women with AAA, who were considered for surgery as part of the Chichester aneurysm screening programme. Sixty-one patients were electively operated and four because of symptoms. The rest were followed up further. Twelve of these experienced ruptured AAA later. RESULTS: No correlation between last measured AAA-diameter, annual expansion rate and SEP was noticed. However, SEP levels were significantly higher in cases rupturing later, persisting after adjustment for age, sex, and last measured AAA-size. ROC curve analysis concerning SEP as a predictor of rupture later showed an optimal sensitivity and specificity of 67% and 60%, respectively, similar with last measured AAA-size. By combining AAA-size and SEP, the optimal sensitivity and specificity reached 83% and 66%, respectively. CONCLUSION: One sampling of SEP combined with AAA-size in patients referred for AAA surgery may be a clinical useful indicator of high rupture risk. PMID- 11735206 TI - Varicose veins: loss of release of vascular endothelial growth factor and reduced plasma nitric oxide. AB - OBJECTIVES: To examine the release of vascular endothelial growth factor (VEGF) and nitric oxide (NO) in primary varicose veins (VVs) and normal vein controls following experimentally-induced venous stasis. DESIGN, MATERIALS AND METHODS: Patients with primary VVs (n=21) and control subjects (n=11) were rested supine for 15 min. Blood was collected from both an arm and foot vein. A below-knee cuff was applied and inflated to 90-95 mmHg for 10 min (to prompt venous stasis). Further blood samples were collected from the foot vein. Levels of plasma VEGF and NO were analysed. RESULTS: In control subjects, application of the cuff increased levels of plasma VEGF in the foot (p<0.025). In contrast, in patients with VVs, there was little or no change in these levels of plasma VEGF. Cuff application had little effect on levels of plasma NO in either controls, or those with VVs. When compared to controls however, the levels of plasma NO in all samples with VVs (arm, or foot before, or after, cuff application) were reduced (all, p<0.05). CONCLUSIONS: Loss of VEGF release with experimentally-induced venous stasis, and reduced levels of plasma NO may suggest a mechanism important in the development of primary VVs. PMID- 11735207 TI - Primary autologous superficial femoral vein reconstruction of an emergency, infected, ruptured aortic aneurysm. PMID- 11735208 TI - Endovascular exclusion of bilateral common iliac artery aneurysms with preservation of internal iliac artery perfusion. PMID- 11735209 TI - Rupture of an abdominal aortic aneurysm secondary to type II endoleak. PMID- 11735210 TI - Athletes with lower limb ischaemia. AB - The case of a young healthy sportsman and acute exacerbation of chronic infragenicular pain is presented. Further investigation revealed an obstruction of the tibiofibular trunk due to an osteochondroma, arising from the fibula, which was immediately resected. Osteochondroma is observed in 1-2% of the population and may present with vascular complications. In young patients and athletes, leg pain may be of vascular origin due to an entrapment or compression and should always be considered. PMID- 11735211 TI - The consequences of an abdominal aortic aneurysm infected with methicillin resistant Staphylococcus aureus (MRSA). PMID- 11735212 TI - Thrombosed abdominal aortic aneurysms. Do they need surveillance to prevent late rupture? PMID- 11735213 TI - Delayed common femoral artery rupture following irradiation therapy for carcinoma of the vulva. PMID- 11735215 TI - "Science" education. PMID- 11735216 TI - Genetic identification of a locus, Mot1, that affects renal tumor size in the rat. AB - Prognosis and treatment of solid tumors are directly dependent on the stage of disease. For any type of cancer, tumor characteristics such as size, multiplicity, and metastatic potential are highly heterogeneous among patients. Our understanding of the genetic determinants of tumor burden is rudimentary. Here, rats carrying a germline mutation of the gene Tsc2 were found to develop variable size and number of renal tumors. We hypothesize that "modifier" genes unlinked to Tsc2 affect its expressivity. Using a backcross (BC) analysis between the two strains that showed the greatest difference in tumor size (Fischer344 and Brown Norway), we mapped a quantitative trait locus based on tumor volume to rat chromosome 3q, lying in the interval between D3Mit3 and D3Rat17, with a maximum lod score of 4.4. This locus, Mot1 (modifier of Tsc2 1), accounts for approximately 35% of the genetic variation in tumor size between the two strains. No significant difference in tumor multiplicity was noted between Brown Norway and Fischer344 rats. This suggests that Mot1 modulates the rate of disease progression and not tumor initiation. Candidate genes on rat chromosome 3 included Tsc1, whose product interacts biochemically with the TSC2 protein, but it was excluded on the basis of linkage analysis (LOD=0.01). Comparative genomics suggest that the Mot1 region is represented by human chromosomes 15q and 20pq. Our results provide the first evidence of a modifier gene affecting the Tsc2 pathway in the progression of renal tumorigenesis. PMID- 11735217 TI - Identification of new regulatory sequences far upstream of the mouse monocyte chemoattractant protein-1 gene. AB - We systematically searched for sequences influencing the expression of the mouse monocyte chemoattractant protein-1 (MCP-1) gene (Scya2) by mapping DNase I hypersensitive sites (HS) in the chromatin of mesangial cells in a 40-kb interval around the gene. We found nine HS located between -24 kb and +12.7 kb. Three HS coincided with previously known regulatory sequences (HS-2.4, HS-1.0, and HS 0.2). We tested two of the previously unknown HS located far upstream of Scya2 (HS-19.4 and HS-16.3) in transfection experiments using luciferase reporter constructs and mouse mesangial cells as recipients. In transient transfections, both HS had a moderate effect on basal promoter activity as well as promoter activity stimulated by tumor necrosis factor-alpha. In stable transfection experiments, we found much higher activity. A DNA fragment containing HS-19.4 and HS-16.3 caused a considerable increase in the number of stably integrated luciferase copies. We determined the nucleotide sequence of the 5' flanking region to -28.6 kb. Computer-assisted sequence analysis did not yield evidence of an additional gene. These HS are located within the 5' flanking region of a gene cluster consisting of Scya2 (MCP-1), Scya7 (MCP-3), Scya11 (eotaxin), Scya12 (MCP 5), and Scya8 (MCP-2). This report represents the first comprehensive chromatin analysis of the mouse MCP-1 locus leading to the identification of a complex regulatory region located far upstream of Scya2. PMID- 11735218 TI - Cloning, pharmacology, and tissue distribution of G-protein-coupled receptor GPR105 (KIAA0001) rodent orthologs. AB - It has recently been shown that UDP-glucose is a potent agonist of the orphan G protein-coupled receptor (GPCR) KIAA0001. Here we report cloning and analysis of the rat and mouse orthologs of this receptor. In accordance with GPCR nomenclature, we have renamed the cDNA clone, KIAA0001, and its orthologs GPR105 to reflect their functionality as G-protein-coupled receptors. The rat and mouse orthologs show 80% and 83% amino acid identity, respectively, to the human GPR105 protein. We demonstrate by genomic Southern blot analysis that there are no genes in the mouse or rat genomes with higher sequence similarity. Chromosomal mapping shows that the mouse and human genes are located on syntenic regions of chromosome 3. Further analyses of the rat and mouse GPR105 proteins show that they are activated by the same agonists as the human receptor, responding to UDP glucose and closely related molecules with similar affinities. The mouse and rat receptors are widely expressed, as is the human receptor. Thus we conclude that we have identified the rat and mouse orthologs of the human gene GPR105. PMID- 11735219 TI - The Stat3/5 locus encodes novel endoplasmic reticulum and helicase-like proteins that are preferentially expressed in normal and neoplastic mammary tissue. AB - The signal transducers and activators of transcription (STAT) 5 and 3 are critical for mammary alveolar development during pregnancy and remodeling during involution. In the mouse, STAT3, STAT5a, and STAT5b are encoded by adjacent genes on chromosome 11 (60.5 cM). To identify additional genes in the Stat3/5 locus that may participate in normal and neoplastic development of the mammary gland, we have cloned and sequenced 500 kb and searched for genes preferentially expressed in mammary tissue. We identified six known genes and cloned two new genes, termed D11Lgp1 and D11Lgp2. Both genes are most highly expressed in normal mammary tissue and mammary tumors from several transgenic mouse models. LGP1 consists of 532 and 530 amino acids in mouse and human, respectively (88% similarity). A region in the carboxy-terminal half of LGP1 has limited homology with Arabidopsis thaliana GH3-like proteins. Immunofluorescence studies demonstrated that LGP1 is located in the nuclear envelope and the endoplasmic reticulum. LGP2 is a cytoplasmic protein of 678 amino acids. PMID- 11735221 TI - Migraine with aura susceptibility locus on chromosome 19p13 is distinct from the familial hemiplegic migraine locus. AB - Migraine is a common neurological disease with a major genetic component. Recently, it has been proposed that a single locus on chromosome 19p13 contributes to the genetic susceptibility of both rare familial hemiplegic migraine (FHM) and more common types of migraine, migraine with aura and migraine without aura. We analyzed 16 families for co-segregation of migraine with aura and chromosome 19p13 markers. Using multipoint model-free linkage analysis, we obtained a lod score of 4.28 near D19S592. Using an affecteds-only model of linkage, we observed a lod score of 4.79 near D19S592. We were able to provide statistical evidence that this locus on chromosome 19p13 is most likely not the gene CACNA1A, mutations in which cause FHM. These data indicate that chromosome 19p13 contains a locus which contributes to the genetic susceptibility of migraine with aura that is distinct from the FHM locus. PMID- 11735220 TI - Single-nucleotide polymorphism alleles in the insulin receptor gene are associated with typical migraine. AB - We have identified a migraine locus on chromosome 19p13.3/2 using linkage and association analysis. We isolated 48 single-nucleotide polymorphisms within the locus, of which we genotyped 24 in a Caucasian population comprising 827 unrelated cases and 765 controls. Five single-nucleotide polymorphisms within the insulin receptor gene showed significant association with migraine. This association was independently replicated in a case-control population collected separately. We used experiments with insulin receptor RNA and protein to investigate functionality for the migraine-associated single-nucleotide polymorphisms. We suggest possible functions for the insulin receptor in migraine pathogenesis. PMID- 11735222 TI - Assembly and annotation of human chromosome 2q33 sequence containing the CD28, CTLA4, and ICOS gene cluster: analysis by computational, comparative, and microarray approaches. AB - Human chromosome 2q33 is an immunologically important region based on the linkage of numerous autoimmune diseases to the CTLA4 locus. Here, we sequenced and assembled 2q33 bacterial artificial chromosome (BAC) clones, resulting in 381,403 bp of contiguous sequence containing genes encoding a NADH: ubiquinone oxidoreductase, the costimulatory receptors CD28, CTLA4, and ICOS, and a HERV-H type endogenous retrovirus located 366 bp downstream of ICOS in the reverse orientation. Genomic microarray expression analysis using differentially activated T-cell RNA against a subcloned CTLA4/ICOS BAC library revealed upregulation of CTLA4 and ICOS sequences, plus antisense ICOS transcripts generated by the HERV-H, suggesting a potential mechanism for ICOS regulation. We identified four nonlinked, polymorphic, simple repetitive sequence elements in this region, which may be used to delineate genetic effects of ICOS and CTLA4 in disease populations. Comparative genomic analysis of mouse genomic Icos sequences revealed 60% sequence identity in the 5' UTR and regions between exon 2 and the 3' UTR, suggesting the importance of ICOS gene function. PMID- 11735223 TI - Comparative genomic sequence analysis of the FXR gene family: FMR1, FXR1, and FXR2. AB - Mutations in the X-linked gene FMR1 cause fragile X syndrome, the leading cause of inherited mental retardation. Two autosomal paralogs of FMR1 have been identified, and are known as FXR1 and FXR2. Here we describe and compare the genomic structures of the mouse and human genes FMR1, FXR1, and FXR2. All three genes are very well conserved from mouse to human, with identical exon sizes for all but two FXR2 exons. In addition, the three genes share a conserved gene structure, suggesting they are derived from a common ancestral gene. As a first step towards exploring this hypothesis, we reexamined the Drosophila melanogaster gene Fmr1, and found it to have several of the same intron/exon junctions as the mammalian FXRs. Finally, we noted several regions of mouse/human homology in the noncoding portions of FMR1 and FXR1. Knowledge of the genomic structure and sequence of the FXR family of genes will facilitate further studies into the function of these proteins. PMID- 11735224 TI - Nucleotide sequence, genomic organization, and chromosomal localization of genes encoding the human NMDA receptor subunits NR3A and NR3B. AB - The N-methyl-D-aspartate (NMDA) receptors are glutamate-regulated ion channels that are critically involved in important physiological and pathological functions of the mammalian central nervous system. We have identified and characterized the gene encoding the human NMDA receptor subunit NR3A (GRIN3A), as well as the gene (GRIN3B) encoding an entirely novel subunit that we named NR3B, as it is most closely related to NR3A (57.4% identity). GRIN3A localizes to chromosome 9q34, in the region 13-34, and consists of nine coding exons. The deduced protein contains 1115 amino acids and shows 92.7% identity to rat NR3A. GRIN3B localizes to chromosome 19p13.3 and contains, as does the mouse NR3B gene (Grin3b), eight coding exons. The deduced proteins of human and mouse NR3B contain 901 and 900 amino acid residues, respectively (81.6% identity). In situ hybridization shows a widespread distribution of Grin3b mRNA in the brain of the adult rat. PMID- 11735225 TI - A family of 12 human genes containing oxysterol-binding domains. AB - Oxysterol-binding proteins (OSBPs) have been described in a wide range of eukaryotes, and are often found to be part of a multi-gene family. We have used bioinformatics and data mining as a starting point for identifying new family members in humans based on the presence of the OSBP signature EQVSHHPP. In addition to OSBP and the recently reported OSBP2, we have found 10 other genes encoding oxysterol-binding domains. Here, we report cDNA and deduced peptide sequences of the previously unknown OSBPs and compare the peptides and genes. All of the genes encode a pleckstrin homology domain, except OSBPL2. However, two of the peptides, OSBPL2 and OSBPL1A, consist of the OSBP domain only. A second OSBPL1 transcript (OSBPL1B) contains 15 additional upstream exons, with a deduced peptide containing a pleckstrin homology domain. Cladistic analysis divides the human OSBP genes into five groups, whose members share similarities in sequence and gene structure; RT-PCR analysis indicates that expression patterns among group members vary widely. PMID- 11735226 TI - A high-resolution genetic, physical, and comparative gene map of the doublefoot (Dbf) region of mouse chromosome 1 and the region of conserved synteny on human chromosome 2q35. AB - The mouse doublefoot (Dbf) mutant exhibits preaxial polydactyly in association with craniofacial defects. This mutation has previously been mapped to mouse chromosome 1. We have used a positional cloning strategy, coupled with a comparative sequencing approach using available human draft sequence, to identify putative candidates for the Dbf gene in the mouse and in homologous human region. We have constructed a high-resolution genetic map of the region, localizing the mutation to a 0.4-cM (+/-0.0061) interval on mouse chromosome 1. Furthermore, we have constructed contiguous BAC/PAC clone maps across the mouse and human Dbf region. Using existing markers and additional sequence tagged sites, which we have generated, we have anchored the physical map to the genetic map. Through the comparative sequencing of these clones we have identified 35 genes within this interval, indicating that the region is gene-rich. From this we have identified several genes that are known to be differentially expressed in the developing mid gestation mouse embryo, some in the developing embryonic limb buds. These genes include those encoding known developmental signaling molecules such as WNT proteins and IHH, and we provide evidence that these genes are candidates for the Dbf mutation. PMID- 11735227 TI - Homology between a 173-kb region from mouse chromosome 10, telomeric to the Ifng locus, and human chromosome 12q15. AB - We sequenced a 173-kb region of mouse chromosome 10, telomeric to the Ifng locus, and compared it with the human homologous sequence located on chromosome 12q15 using various sequence analysis programs. This region has a low density of genes: one gene was detected in the mouse and the human sequences and a second gene was detected only in the human sequence. The mouse gene and its human orthologue, which are expressed in the immune system at a low level, produce a noncoding mRNA. Nonexpressed sequences show a higher degree of conservation than exons in this genomic region. At least three of these conserved sequences are also conserved in a third mammalian species (sheep or cow). PMID- 11735228 TI - Characterization of a mouse gene (Fbxw6) that encodes a homologue of Caenorhabditis elegans SEL-10. AB - The SCF complex is a type of ubiquitin ligase that consists of the invariable components SKP1, CUL1, and RBX1 as well as a variable component, known as an F box protein, that is the main determinant of substrate specificity. The Caenorhabditis elegans F-box- and WD40-repeat-containing protein SEL-10 functionally and physically associates with LIN-12 and SEL-12, orthologues of mammalian Notch and presenilin, respectively. We have now identified a gene (which we call Fbxw6) that encodes a mouse homologue (F-box-WD40 repeat protein 6, or FBW6) of SEL-10 and is expressed mainly in brain, heart, and testis. Co immunoprecipitation analysis showed that FBW6 interacts with SKP1 and CUL1, indicating that these three proteins form an SCF complex. Comparison of the genomic organization of Fbxw6, which is located on mouse chromosome 3.3E3, with that of mouse Fbxw1, Fbxw2, and Fbxw4 showed only a low level of similarity, indicating that these genes diverged relatively early and thereafter evolved independently. PMID- 11735229 TI - RGSZ1 and Ret RGS: two of several splice variants from the gene RGS20. AB - RGSZ1 and Ret RGS, members of the regulator of G-protein signaling (RGS) family, are GTPase-activating proteins (GAPs) with high selectivity for G alpha(z). We show here that RGSZ1 and Ret RGSZ1 are products of two of several splice variants of one gene, RGS20. RGS20 spans approximately 107 kb and contains at least seven exons. Five exons account for RGSZ1, including a single exon distinct to RGSZ1 that encodes a newly identified amino-terminal region. The previously described open reading frame (ORF) and 3' untranslated region are encoded by four downstream exons that also encode about half of Ret RGS. The 5' end of the RGSZ1 ORF contains several in-frame ATG codons (3-5 depending on the species), and multiple translational start sites may help explain the molecular weight heterogeneity of purified bovine brain RGSZ. Ret RGS replaces the 24 N-terminal amino acid residues of RGSZ1 with a large, N-terminal region that initially distinguished the bovine Ret RGS from human and mouse RGSZ1. This N-terminal domain is encoded by two distinct 5' exons that are variably combined with the four downstream exons shared with RGSZ1 to produce at least six mRNAs. They encode proteins with N termini that vary in size, hydrophobicity, and the presence of a cysteine string. At least two mRNAs that include the exon that encodes the N-terminal region unique to RGSZ1 were found in brain and a few other tissues, but not retina. RGS20 thus can account for multiple G(z)-selective GAPs in different tissues. PMID- 11735230 TI - Diagnosis and management of growth hormone deficiency in childhood and adolescence. Part 1: diagnosis of growth hormone deficiency. PMID- 11735231 TI - Safety aspects of pharmacological GH therapy in adults. AB - The consequences of "pharmacological" growth hormone administration have been studied in a number of conditions, including those characterized by high rates of catabolism. The majority of studies have reported favourable effects on metabolism but recent reports indicate that GH treatment results in increased mortality in critically ill humans. The objective of the study was to assess the safety of large doses of rhGH therapy in human adults. Original trials were identified by searching MEDLINE (1966-March 2000) and the Cochrane database (2000). References of all identified trials were also inspected for more studies. All relevant trials in which GH had been administered to non-GH-deficient (GHD) adult humans were selected from. Outcomes such as death, clinically significant change in function, change in length of hospital stay or need for treatment, and adverse effects were sought. Studies were selected, quality-assessed and passed suitable for inclusion by two independent reviewers. Those studies that were placebo-controlled with satisfactory randomization were considered for inclusion. Twenty-one reports were included in the review. A wide range of patient groups were studied by a variety of investigators, employing a range of doses and duration of GH treatment. The study protocols differed markedly. The majority of studies were small and were designed and/or powered to enable identification of specific effects on nutritional status, protein metabolism, level of function or quality of life. Only two studies were designed to assess safety issues and mortality. In these, GH treatment was associated with a marked increase in mortality in critically ill ICU patients, with a range of diagnoses. Multi-organ failure and the effects of sepsis/infection accounted for most of the excess mortality. In addition morbidity, in terms of length of ICU stay, was increased by GH administration. Other less marked effects were increased fluid retention and hyperglycaemia as a consequence of GH administration. Functional improvement following GH therapy was documented in some studies. There have been few studies assessing the safety aspects of "pharmacological" GH treatment in adult humans. Two well-designed reports indicate that GH administration results in increased morbidity and mortality in a wide variety of critically ill subjects across a spectrum of age ranges. The mechanism(s) of the GH-associated mortality remain poorly understood. Based on current trial evidence, pharmacological GH treatment cannot be recommended for widespread use in critically ill subjects. Well conducted and reported randomized trials are still needed to inform practice as to whether GH administration will be safe in specific illness categories. PMID- 11735232 TI - The use of pigs as an animal model to evaluate the efficacy, potency and specificity of two growth hormone releasing factor analogues. AB - In 1982, Guillemin et al reported the isolation of the human (h) growth hormone (GH) releasing factor (GRF) from a pancreatic tumour in an acromegalic patient. Since then, work to develop potent GRF analogues has been widespread and the rat has been the main animal model used. The aim of the present study was to compare the efficacy, potency and specificity of two GRF analogues with those of the native GRF(1-29)NH(2)using pig (p) as the animal model. Two analogues, Al ([His(1), D-Ala(2), Ala(8,9,15,17), D-Arg(29)] hGRF(1-29)NH(2)) and A2 ([D Ala(2), Ala(8,9,15,17), D-Arg(29)] hGRF(1-29)NH(2)) were compared with the h or pGRF(1-29)NH(2). Five studies were designed using 28-48 kg BW growing barrows. Results showed that the two GRF analogues were more potent than the native GRF molecule, were highly specific, were active for long periods of time and were able to induce changes in body composition similar to those reported with GH or other GRF analogues. Because of the similarity between swine and human species with respect to the amino acid sequence of GRF and to the physiology, secretion and effects of GH, it can be proposed that the pig could be used as a pre clinical animal model to study and test new GRF molecules over short and long periods of time. PMID- 11735233 TI - Similar cardiovascular effects of growth hormone and insulin-like growth factor-I in rats after experimental myocardial infarction. AB - Accumulating data show that growth hormone (GH) and insulin-like growth factor-I (IGF-I) have major effects on the cardiovascular system. In the present study we have directly compared GH and IGF-I in an in vivo rat model of experimental myocardial infarction. Four weeks after ligation of the left coronary artery, male rats were treated with recombinant human (rh) GH 1.1 mg/kg per day, rhIGF-I 3.0 mg/kg per day or saline s.c. for 2 weeks. Transthoracic echocardiography was performed before and after the treatment period. Both GH and IGF-I reduced total peripheral resistance (P< 0.01), end-systolic wall stress (P< 0.01) and end systolic short-axis area (P< 0.001 and P< 0.05). GH also increased area fractional shortening (P< 0.05). Stroke volume (SV) and SV index were improved by IGF-I (P< 0.0001), and SV tended to be increased by GH (P= 0.12). In conclusion, GH and IGF-I had similar beneficial effects on systolic function and peripheral resistance after experimental myocardial infarction. PMID- 11735234 TI - Growth hormone treatment and type 2 diabetes. PMID- 11735235 TI - Growth hormone signalling and its regulation: preventing too much of a good thing. AB - Growth hormone (GH) is a major growth-promoting and metabolic regulatory hormone. Interaction of GH with its cell surface GH receptor (GHR), by virtue of receptor dimerization, causes activation of the GHR-associated cytoplasmic tyrosine kinase, JAK2. Several signalling pathways, including the STAT5, PI3 kinase and MAP kinase pathways, are thereby accessed, resulting in various biochemical and biological cellular signalling outcomes which are rapidly becoming deciphered. Various mechanisms probably exist to terminate, modulate and prevent GH signalling. Some of these mechanisms regulate receptor abundance and/or availability while others may alter the responsiveness of downstream signalling molecules to receptor engagement. In this review, recent insights into modulation of GH signalling are discussed. Special emphasis is placed on mechanisms of homologous and heterologous desensitization and on the likelihood that inducible GHR proteolysis, in addition to causing GH binding protein generation, may also serve as an important mechanism of heterologous GHR downregulation. PMID- 11735236 TI - Local IGF-I expression and bone formation. PMID- 11735237 TI - Granulosa cell proliferation is impaired in the Igf1 null ovary. AB - Insulin-like growth factor-I (IGF-I) expression is highly correlated with ovarian follicular growth and granulosa cell proliferation in both pre-pubertal and mature murine ovaries. Igf1 gene deleted mice are infertile, with ovarian follicles arrested at an early stage of development. To elucidate the cause of follicular dysfunction in Igf1 null mice, this study compared granulosa cell proliferation at baseline and in response to exogenous oestradiol (E2) in prepubertal Igf1 null and wild-type (WT) littermate mice. The basal granulosa cell mitotic index was 3.8+/-0.48 in WT and 1.3+/-0.7 in Igf1 null mice (P=0.03). After E2 treatment, WT granulosa mitotic index was 12.7+/-0.0 vs 5.5+/-0.8 for Igf1 null mice (P<0.001). Granulosal BRDU incorporation was also significantly reduced as were cyclin D2 and B1 immunoreactivities in Igf1 null compared with WT mice. The incidence of apoptosis was not increased in Igf1 null follicles, although BAX immunostaining was increased. These data suggest that IGF1 is essential for normal basal and oestrogen-induced granulosa cell proliferation and follicular growth. PMID- 11735238 TI - A study of the growth-promoting and metabolic effects of growth hormone (GH) in a patient with the "growth without GH" syndrome. AB - The paradox of normal or even excessive growth despite a proven lack of GH is a well-known but still unexplained phenomenon that has been described in some patients following resection of a craniopharyngioma or other suprasellar tumours. However, the consequences of GH deficiency on other metabolic aspects of GH action in this syndrome have not been adequately investigated. The aim of this study was to examine whether a dissociation might exist between the growth promoting and metabolic effects of GH. We studied a 7.1 year old boy who, after removal of a suprasellar craniopharyngioma, developed panhypopituitarism with mild hyperprolactinaemia. Despite the presence of severe GH deficiency associated with persistently low IGF-I and IGFBP-3 levels, the patient grew spontaneously at an accelerated rate for a prepubertal boy, achieving a height velocity of 9.0 cm during the first and 8.5 cm during the second post-operative year. However, other metabolic parameters of GH activity were adversely affected by the lack of GH. The maximum tubular reabsorption rate for phosphate over glomerular filtration rate ratio (2.8) was persistently low and normalized during a 4 day course of hGH administration (4.2) together with the normalization of IGF-I (from 34 microg/l to 294 microg/l), suggesting that GH-dependent renal phosphate handling is impaired in this syndrome. In addition, bone age was delayed by 1.7 years consistently with delayed skeletal maturation, whereas skinfold thickness and the waist to hip ratio were increased in comparison with normative data, suggesting increased adipose tissue mass with central fat distribution, a phenotype characteristic of GH deficiency. In conclusion, our case study suggests that, in the "growth without GH" syndrome, the excessive growth is independent of GH and dissociated from other GH-dependent metabolic effects, which are decreased. PMID- 11735239 TI - Effect of IGFBP-3 on IGF- and IGF-analogue-induced insulin-like growth factor-I receptor (IGFIR) signalling. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) binds IGF-I and IGF-II with high affinity, at least an order of magnitude higher than the affiniy of the IGFs for the IGFIR. It has been hypothesized that IGFBP-3 inhibits IGF binding to the IGFIR via a mechanism independent of its ability to sequester IGFs. In the present study, we examined the effects of IGFBP-3 and its proteolytic fragments on the initial events of the IGFIR signalling pathway. IGFBP-3 inhibited IGF-I-, IGF-II-, Des(1-3)IGF-I- and Long(R3)IGF-I-induced IGFIR phosphorylation in a dose dependent manner at similar concentration range but not QAYL-induced IGFIR-P. The((1-97))IGFBP-3 fragment was able to inhibit only IGF-I-induced IGFIR-P. The((1-97))IGFBP-3 fragment but not intact IGFBP-3 inhibited insulin-induced IGFIR-P. Monolayer cross-linking with [(125)I]IGFBP-3 indicated that there is no direct interaction of IGFBP-3 with the IGFIR. This study demonstrates that the effect on the initial step of IGFIR signalling by IGFBP-3 is largely due to its ability to sequester IGF and the IGF analogues in the extracellular milieu and not the result of any interaction of IGFBP-3 with the IGFIR or a mechanism independent of its ability to bind IGFs. PMID- 11735240 TI - Effects of testosterone either alone or with IGF-I on growth of cells derived from the proliferation zone of regenerating antlers in vitro. AB - Deer antlers are male secondary sexual characters and are the fastest growing mammalian tissue. As such, both androgens and growth factors play a major role in antler development. The timing of the antler cycle is controlled by the seasonal fluctuations of testosterone, and the actual growth of antlers is mainly stimulated by growth factors including insulin-like growth factor-1 (IGF-I). However, whether or not testosterone at low levels plays a growth-promoting role during antler formation is controversial. In the present study, we took an in vitro approach to investigate whether testosterone either alone or with IGF-I had mitogenic effects on mesenchymal or cartilaginous cells derived from the proliferation zone of regenerating antlers. In addition, a binding assay was carried out to determine whether the specific binding sites for testosterone were preserved after cell disaggregation. The results showed that testosterone either in physiological concentrations or at low levels did not exert direct mitogenic effects on antler cells derived from the proliferation zone in serum-free medium in vitro (P>0.05), even if the specific binding sites for testosterone in these cells were well preserved. Likewise, testosterone in a very wide range of concentrations not only failed to enhance (P>0.05), but at certain levels (0.1-5 nM) impaired the mitogenic effects of IGF-I on these antler cells in vitro (P<0.001). Therefore, these results support neither a conclusion that low level testosterone has growth-promoting effects on antler formation nor the hypothesis that testosterone effects may be achieved through sensitizing these antler cells to the mitogenic effects of IGF-I. PMID- 11735241 TI - Severe obstructive sleep apnoea syndrome in an adult patient with Laron syndrome. AB - A 68 year old patient with Laron syndrome (primary growth hormone (GH) resistance insensitivity due to a molecular defect of the GH receptor) and severe obstructive sleep apnoea syndrome is described. Treatment with continuous positive air pressure therapy resulted in improved nocturnal sleep, daytime alertness and cognitive functions. PMID- 11735243 TI - Editorial review: the case of local versus endocrine IGF-I actions: the jury is still out. PMID- 11735242 TI - Tissue-specific regulation of IGF-I and IGF-binding proteins in response to TNFalpha. AB - The circulating concentration of insulin-like growth factor-I (IGF-I) is regulated by both its rate of synthesis and its ability to form stable complexes with IGF-binding proteins (IGFBPs). An equilibrium between IGF-I and IGFBPs is thought to help maintain muscle protein balance. In contrast, catabolic conditions disrupt the IGF system and result in the loss of skeletal muscle protein. We have examined the mechanisms by which tumour necrosis factor alpha (TNFalpha), a catabolic cytokine, alters the IGF system. Conscious rats were infused intravenously with recombinant human TNFalpha or vehicle for 24 h. TNFalpha decreased the concentration of both total and free IGF-I in the plasma (30-40%). This change was associated with a reduction in IGF-I mRNA expression in liver (39%), gastrocnemius (73%), soleus (46%) and heart (63%), but a 2.5-fold increase in the whole kidney. In contrast, TNFalpha did not alter IGF-II mRNA expression in skeletal muscle. TNFalpha also increased IGFBP-1 in the blood (4 fold) and this response was associated with an increase in IGFBP-1 mRNA expression in both liver (3-fold) and kidney (9-fold). In contrast, IGFBP-3 levels in the blood were reduced 38% in response to the infusion of TNFalpha. This change was accompanied by a 60-80% reduction of IGFBP-3 mRNA in liver and kidney but no significant change in muscle. Hepatic mRNA levels of the acid labile subunit were also reduced by TNFalpha (46%). Finally, tissue expression of mac25 (also referred to IGFBP-related protein-1) mRNA was increased in gastrocnemius (50%) but remained unchanged in liver and kidney. These results more fully characterize the changes in various elements of the IGF system and, thereby, provide potential mechanisms for the alterations in the circulating IGF system as well as for changes in tissue metabolism observed during catabolic insults associated with increased TNFalpha expression. PMID- 11735244 TI - The growth hormone secretagogue ipamorelin counteracts glucocorticoid-induced decrease in bone formation of adult rats. AB - The ability of the growth hormone secretagogue (GHS) Ipamorelin to counteract the catabolic effects of glucocorticoid (GC) on skeletal muscles and bone was investigated in vivo in an adult rat model. Groups of 8-month-old female rats were injected subcutaneously for 3 months with GC (methylprednisolone) 9 mg/kg/day or GHS (Ipamorelin) 100 microg/kg three times daily, or both GC and GHS in combination. The maximum tetanic tension of the calf muscles was determined in vivo in a materials testing machine. The maximum tetanic tension was increased significantly, and the periosteal bone formation rate increased four-fold in animals injected with GC and GHS in combination, compared with the group injected with GC alone. In conclusion, the decrease in muscle strength and bone formation found in GC-injected rats was counteracted by simultaneous administration of the growth hormone secretagogue. PMID- 11735245 TI - Commencing growth hormone replacement in adults with a fixed low dose. Effects on serum lipoproteins, glucose metabolism, body composition, and cardiovascular function. AB - The safety and effects of a fixed low dose of growth hormone (GH), 0.17 mg/day was evaluated for 3 months, on glucose metabolism, serum lipids, body composition and cardiac function in 53 GH deficient adults aged 18-78 years. Body composition was determined by dual energy X-ray absorptiometry and total body water was determined by bioelectrical impedance. Echocardiography was used to assess cardiac function and bicycle ergonometry was used to determine exercise capacity. All investigations were performed at baseline and after 3 months. At 3 months, serum levels of insulin-like growth factor (IGF)-I, IGF binding protein (IGFBP)-3 and lipoprotein (a) and lean body mass were increased (P<0.05). Total and low density lipoprotein cholesterol levels and fat mass were reduced (P<0.05). There was an increase in the serum glucose value at 120 min after an oral glucose tolerance test performed at 3 months (P<0.05), no other changes in glucose metabolism or in cardiac function were noted. Side-effects were few and mild. This fixed low-dose regime resulted in improvements in body composition and lipid profile, without causing serious side effects. This is therefore a valid method to institute GH replacement in adults. PMID- 11735246 TI - Regulation of GHRH receptor gene expression in the neonatal and adult rat pituitary. AB - The growth hormone releasing hormone (GHRH) receptor gene is essential for normal growth, and its expression is developmentally regulated. The factors that control GHRH receptor expression in the neonatal pituitary are not well understood. This study focuses on the regulation of GHRH receptor gene expression by thyroid hormone, glucocorticoids, insulin-like growth factor-I (IGF-I) and IGF-II in rat pituitary cell cultures. In newborn pituitaries, both T3 and hydrocortisone (24 h) caused a dose-dependent increase in GHRH receptor mRNA abundance, reaching levels 4.8-fold (P<0.001) and 6.5-fold (P<0.001) over corresponding controls. T3 and hydrocortisone also stimulated GHRH receptor expression in adult (70 day) pituitary cell cultures, consistent with our earlier findings. IGF-I treatment suppressed the inductive effects of T3 (P<0.02) and hydrocortisone (P<0.03) on GHRH receptor expression in adult pituitaries but not in neonatal pituitaries. Unlike IGF-I, IGF-II treatment had no effect on T3-induced or hydrocortisone induced GHRH receptor expression in either neonates or adults. Taken together, these results indicate that (1) thyroid hormone and hydrocortisone act directly at the neonatal pituitary as potent stimulators of GHRH receptor gene expression, (2) IGF-I, but not IGF-II, acts at the pituitary to suppress GHRH receptor mRNA expression and (3) the effects of IGF-I on GHRH receptor gene expression are developmentally determined. PMID- 11735247 TI - Transcriptional regulation of IGF-I receptor gene expression by the PAX3-FKHR oncoprotein. AB - Chromosomal translocations that disrupt the molecular organization of transcription factors are typical of a variety of solid and hematopoietic cancers. Alveolar rhabdomyosarcoma (ARMS), a paediatric soft tissue malignant tumour, is characterized by the recurrent translocation t(2;13)(q35;q14) that fuses the 5' DNA binding domain-encoding sequences of the Pax3 gene with the 3' sequences of the FKHR gene. The insulin-like growth factor (IGF) system has an important role in muscle development as well as in the aetiology of paediatric sarcomas, including ARMS. In the present study the potential regulation of the IGF-I receptor (IGF-I-R) gene by PAX3-FKHR at the transcriptional level was investigated. PAX3-FKHR was able to transactivate the IGF-I-R promoter in sarcoma derived cell lines, whereas PAX3 exhibited a reduced potency in comparison to the fusion protein. Furthermore, transfection of the chimera induced a significant increase in the endogenous levels of IGF-I-R protein, suggesting that the IGF-I-R gene is a physiologically-relevant molecular target for the PAX3-FKHR oncogene. PMID- 11735248 TI - Normal size of the lumen of the aorta in dwarf mice lacking IGF-II. AB - The peptide insulin-like growth factor II (IGF-II) is a foetal promoter of growth and differentiation. Genetically modified mice have provided insights into the contribution of IGF-II to the determination of the size of the body and individual organs. Nevertheless, the role of IGF-II in regulating the size of major blood vessels is poorly understood. Here, the transverse luminal and medial area of the aorta and the number of medial cell nuclei are shown not to be significantly different in adult wild-type and Igf2 nullizygous mice, despite a approximately 30% reduction in BW in the latter group. The observations in this study suggest that IGF-II is not an indispensable factor in the determination of the size of the aortic lumen, and are consistent with the view that growth factor redundancy may be a safeguard mechanism to maintain the size of the aorta within a range compatible with viability. PMID- 11735249 TI - Responses of insulin-like growth factor (IGF)-I and IGF-binding proteins to nutritional status in peroxisome proliferator-activated receptor-alpha knockout mice. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) plays a central role in glucose and lipid homeostasis. Mice lacking PPARalpha(-/-) have a sexually dimorphic phenotype. We have characterized the IGF system in wild type and PPARalpha-/- mice. In normal mice fasting IGF-I and the IGFBP-3 ternary complex were 2-fold higher in males than in females. PPARalpha influenced the IGF/IGFBP response to feeding, particularly in males. Compared to wild type, male PPARalpha /- mice had 40% lower total fasting IGF-I concentrations, decreased ALS and less IGFBP-3 ternary complex formation, but within 4 h of refeeding there was an increase in IGF-I and IGFBP-3 ternary complex to values similar to controls. Circulating IGFBP protease activity was induced in male PPARalpha-/- mice during refeeding. IGFBP-1 and insulin concentrations were higher in males than females, and were increased by PPARalpha knockout, suggesting significant hepatic insulin resistance. We speculate that gender differences in the IGF system contribute to the PPARalpha-/- phenotype. PMID- 11735250 TI - The effect of growth hormone on insulin-like growth factor I and bone metabolism in distraction osteogenesis. AB - Limb lengthening in the left tibia of 30 mature female Yucatan micropigs was performed using distraction osteogenesis. A treatment group of 15 animals received recombinant porcine growth hormone (r-pGH) (100 microg/kg/day) while the others served as controls. Serial serum measurements of total insulin-like growth factor I (IGF-I), free IGF-I, IGF binding proteins -1, -2, -3 and -4 (IGFBP-1 to 4) were performed. Bone-specific alkaline phosphatase (bone-ALP) and the serum carboxyl-terminal telopeptide of type I collagen (ICTP) were measured as bone turnover markers. The GH-treated animals showed a significant increase in total IGF-I, free IGF-I and IGFBP-3 after surgery (P<0.001). Similarly, the treated animals showed a significantly higher level of bone-ALP (P<0.001) throughout the experiment compared to the controls. There was a significant correlation between bone-ALP and total IGF-I (r=0.76) in the GH-treated group and an even higher correlation for free IGF-I (r=0.90). There was no difference in the ICTP serum levels between the two groups. These data indicate that the application of species-specific growth hormone results in a stimulation of bone formation in distraction osteogenesis which may be mediated by IGF-I. The stronger correlation between free IGF-I and bone-ALP indicates that the anabolic effect of IGF-I may be regulated through the IGFBPs by binding and inactivating IGF-I. PMID- 11735251 TI - Relative localization of the prolactin receptor binding sites for lactogenic hormones. AB - A monoclonal antibody termed MAb R7B4, directed to an epitope present in prolactin receptors (PRLRs), was used as a tool to map the receptor binding sites for human growth hormone (hGH), ovine prolactin (oPRL) and human placental lactogen (hPL). Although the three hormones completely inhibited the binding of each other to Nb2 cells or rat liver receptors, MAb R7B4 behaviour was different depending on the hormone tested and the receptor source. According to the MAb effects, PRLR from Nb2 cells would locate both hGH and oPRL close to R7B4 epitope, whereas hPL would bind far from the MAb binding site. On the other hand, PRLR from rat liver should bind hGH close to the R7B4 epitope but oPRL and hPL would be recognized by a separate region of the same receptor. Thus, results presented in this paper suggest that PRLR binding sites for hGH, oPRL and hPL do not exactly overlap in spite of full competition between ligands. PMID- 11735252 TI - The last frontier in cardiovascular health: a landmark lecture for the XVII World Congress of the International Society for Heart Research. AB - Following World War II, Vannevar Bush described science as an "endless frontier" that should be made accessible to all Americans. Since then, cardiovascular health has improved markedly, largely because substantial investments in biomedical research led to numerous therapies and prevention strategies for cardiovascular disease. Despite these advances, however, science remains an endless frontier and we continue to face an infinite array of opportunities for improving cardiovascular health. A standard definition for "frontier" is the "farthermost limit of knowledge or achievement". The limits of our knowledge are expanding at an ever accelerated pace. Unfortunately, we do not always apply what we know, and therefore fail to achieve all we could. For example, we have known for two decades that heart attack patients benefit from beta-blockers, but even today, the drugs are not always prescribed. And, health disparities continue to exist among races and communities. Therefore, the "last frontier of cardiovascular health" is the translation and application of our knowledge to improve the cardiovascular health of all people. We will not reach the farthermost limit of achievement without new knowledge. But, in our zeal to expand our knowledge of cardiovascular diseases, we must remember to ensure that what we learn is rapidly applied to improve cardiovascular health. PMID- 11735253 TI - Reperfusion arrhythmias: new insights into the role of the Na(+)/Ca(2+) exchanger. PMID- 11735254 TI - Stress protein involvement in cardioprotection induced by hypothermia. PMID- 11735255 TI - Gates of fate. PMID- 11735256 TI - Time course of atrial fibrillation-induced cellular structural remodeling in atria of the goat. AB - BACKGROUND: Previously we documented cellular structural changes of a non degenerative nature in atrial myocytes after atrial fibrillation (AF) in the goat. The time course of these changes was not studied. METHODS AND RESULTS: Cellular structural changes were studied by light- and electron microscopy and immunohistochemistry in goat atria after 0-16 weeks AF. The first sign of cellular structural remodeling was a more homogeneous chromatin distribution, at 1 week of AF. Sub-structural changes in mitochondria and sarcoplasmic reticulum occurred gradually. Cellular degeneration was absent. The degree of myolysis and glycogen accumulation increased till 8 weeks of AF and did not increase further from thereon. After 16 weeks of AF, 42% of the myocytes in the right atrial free wall were affected by myolysis. The diameter of the atrial myocytes increased. Dedifferentiation of the atrial myocytes was suggested by altered expression patterns of structural proteins, such as the disappearance of cardiotin (1 week), the A-I junctional part of titin (4 weeks), desmin at the intercalated disk (ID) (8 weeks) and a gradual re-expression of alpha-smooth muscle actin. CONCLUSION: Remodeling of the cellular ultrastructure in atrial myocardium of the goat develops progressively during AF. Re-expression of fetal proteins indicate dedifferentiation of atrial myocytes, analogous to observations in hibernating myocardium of the ventricle. PMID- 11735257 TI - Functional consequences of the mutations in human cardiac troponin I gene found in familial hypertrophic cardiomyopathy. AB - Functional consequences of the six mutations (R145G, R145Q, R162W, DeltaK183, G203S, K206Q) in cardiac troponin I (cTnI) that cause familial hypertrophic cardiomyopathy (HCM) were studied using purified recombinant human cTnI. The missense mutations R145G and R145Q in the inhibitory region of cTnI reduced the intrinsic inhibitory activity of cTnI without changing the apparent affinity for actin. On the other hand, the missense mutation R162W in the second troponin C binding region and the deletion mutation DeltaK183 near the second actin tropomyosin region reduced the apparent affinity of cTnI for actin without changing the intrinsic inhibitory activity. Ca(2+) titration of a fluorescent probe-labeled human cardiac troponin C (cTnC) showed that only R162W mutation impaired the cTnC-cTnI interaction determining the Ca(2+) affinity of the N terminal regulatory domain of cTnC. Exchanging the human cardiac troponin into isolated cardiac myofibrils or skinned cardiac muscle fibers showed that the mutations R145G, R145Q, R162W, DeltaK183 and K206Q induced a definite increase in the Ca(2+)-sensitivity of myofibrillar ATPase activity and force generation in skinned muscle fibers. Although the mutation G203S also showed a tendency to increase the Ca(2+) sensitivity in both myofibrils and skinned muscle fibers, no statistically significant difference compared with wild-type cTnI could be detected. These results demonstrated that most of the HCM-linked cTnI mutations did affect the regulatory processes involving the cTnI molecule, and that at least five mutations (R145G, R145Q, R162W, DeltaK183, K206Q) increased the Ca(2+) sensitivity of cardiac muscle contraction. PMID- 11735258 TI - Glutamate-loading stimulates metabolic flux and improves cell recovery following chemical hypoxia in isolated cardiomyocytes. AB - The amino acid glutamate is used in cardioplegic solutions, yet evidence is conflicting as to whether or not exogenous glutamate is indeed cardioprotective. This controversy may be because increasing extracellular glutamate does not necessarily lead to an increase in intracellular glutamate. In this study we aimed to determine whether isolation of myocytes in the presence of glutamate resulted in glutamate-loading of the cells, and, if so, whether such loading protected myocytes from simulated (chemical) hypoxia. Single ventricular myocytes were isolated from rat hearts in the presence and absence of 6.4 mM glutamate. Levels of glutamate and ATP were determined using HPLC, and NADH/NAD(+) was determined from cell autofluorescence. Chemical hypoxia was induced by superfusion with a solution containing 2.5 mM cyanide and no glucose. Intracellular [Ca(2+)] was measured by loading cells with indo-1, and cell length was measured using an edge-tracking device. Isolation of myocytes in the presence of glutamate resulted in increased intracellular glutamate levels compared with cells isolated in the absence of glutamate, 1324+/-108 v 948+/-124 pmol/mg protein, respectively (P<0.05). Cells loaded with glutamate showed increased NADH/NAD(+), (0.384+/-0.032 v 0.281+/-0.029, P<0.05) and greater ATP levels (36.031+/-1.633 nmol/mg protein v 19.279+/-3.327 nmol/mg protein, P<0.005) compared to control cells. When subjected to chemical hypoxia, cells underwent rigor-contracture at various timepoints, and were then reperfused following 5 min in rigor. Cells loaded with glutamate showed better recovery of diastolic [Ca(2+)], Ca(2+) transient amplitude, and improved contractile function compared with cells isolated in absence of glutamate. This study demonstrates an efficient method for loading myocytes with glutamate during cell isolation, and myocytes loaded with glutamate showed increased metabolic flux, as indexed by a higher NADH/NAD(+) and ATP content. Myocytes also exhibited better recovery from chemical hypoxia in terms of both Ca(2+) handling and cell contraction. PMID- 11735259 TI - Real-time RT-PCR for the detection of beta-adrenoceptor messenger RNAs in small human endomyocardial biopsies. AB - Quantification of mRNAs from extremely small human samples remains a challenge. Requiring minimal amounts of tissue and no post-reaction manipulation, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) is an attractive method to quantitatively assess the expression of rare mRNAs. We evaluated the applicability of the technique on RNA extracted from human endomyocardial biopsies and isolated cardiomyocytes, and compared the technique to the RT competitive PCR approach. Primers and probes were designed to amplify the three subtypes of human beta -adrenoceptors (beta1-, beta2- and beta3 AR), as well as reference genes such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Hypoxanthine-guanine phosphoribosyltransferase (HPRT), and the oncogene ABL by real-time RT-PCR. Specific primers and a deleted competitor were synthetized to compare the quantitation of the beta 3 AR mRNA expression by RT-competitive PCR. We validated the technique on human cardiomyocytes either freshly isolated or selectively excised from fixed sections of human myocardium by Laser Capture Microdissection. The standard curves obtained for the cDNA's analysed showed mean slopes comprised between -3.3 and -3.7. Inter- and intra-assay variability of gene quantitation was reflected by mean values of the variance coefficients of Ct of 4.84+/-1.13% and 2.73+/-0.39% or 3.32+/-1.03% and 2.21+/-0.24% (corresponding to percent variances of copy numbers of 83.07+/-12.72% and 34.45+/-9.03% or 47.40+/-8.59% and 23.83+/-3.16%) for human beta3 AR and GAPDH genes, respectively. The expression of GAPDH, HPRT and ABL mRNA was characterized by a very low dispersion of individual values across cardiac pathologies, suggesting that these genes may be used as reference genes in quantitative PCR studies. Finally, we applied the technique to detect rare mRNAs, such as beta -AR mRNAs, from small human endomyocardial biopsies and even isolated cardiomyocytes. Real time RT-PCR is appropriate to quantitate rare messenger RNAs, including in extremely small human tissue samples. This method appears very promising for futures studies of gene expression in several pathophysiological conditions, including heart failure. PMID- 11735260 TI - Regulation of sodium-calcium exchange and mitochondrial energetics by Bcl-2 in the heart of transgenic mice. AB - Our previous work in cultured cells has shown that the maintenance of mitochondrial Ca(2+) homeostasis is essential for cell survival, and that the anti-apoptotic protein Bcl-2 is able to maintain a threshold level of mitochondrial Ca(2+) by the inhibition of permeability transition. To test whether Bcl-2 also affects the mitochondrial Na(+)-Ca(2+) exchange (NCE), a major efflux pathway for mitochondrial Ca(2+), studies using transgenic mice that overexpress Bcl-2 in the heart have been performed. NCE activity was determined as the Na(+)-dependent Ca(2+) efflux in the isolated mitochondria. Overexpression of Bcl-2 led to a significant reduction of NCE activity as well as increased resistance to permeability transition in the mitochondria of transgenic heart. This was accompanied by increased matrix Ca(2+) level, enhanced formation of NADH and enhanced oxidation of pyruvate, an NAD(+)-linked substrate. Furthermore, there was induction of cellular Ca(2+) transport proteins including the Na(+) Ca(2+) exchanger of the sarcolemma (NCX). Bcl-2 not only stimulates NCX expression in the sarcolemma but also attenuates the Na(+)-Ca(2+) exchange in the mitochondria. These results are consistent with the protection by Bcl-2 against apoptosis in heart following ischemia/reperfusion. PMID- 11735261 TI - Activation of connexin-43 hemichannels can elevate [Ca(2+)]i and [Na(+)]i in rabbit ventricular myocytes during metabolic inhibition. AB - ATP depletion due to ischemia or metabolic inhibition (MI) causes Na(+) and Ca(2+) accumulation in myocytes, which may be in part due to opening of connexin 43 hemichannels. Halothane (H) has been shown to reduce conductance of connexin 43 hemichannels and to protect the heart against ischemic injury. We therefore investigated the effect of halothane on [Ca(2+)]i and [Na(+)]i in myocytes during MI. Isolated rabbit left ventricular myocytes were loaded with 4 microM fluo-3 AM for 30 min, or with 5 microM sodium green AM for 60 min at 37 degrees C. After washing, the myocytes were exposed to: (1) Normal HEPES solution; (2) MI solution (2 mM NaCN, 20 mM 2-deoxy-D-glucose and 0-glucose); or (3) MI+H (0.95 mM, 4.7 mM) for 60 min. Propidium iodide (PI, 25 microM) was added to all samples before data acquisition. The fluorescence intensity was measured by flow cytometry with 488 nm excitation and 530 nm emission for fluo-3 or sodium green, and 670 nm for PI. The [Ca(2+)]i and [Na(+)]i were then calculated by calibration. In some experiments, the effect of 10 microM tetrodotoxin (TTX) and 20 microM nifedipine (NIF) were studied. Metabolic inhibition for 60 min caused a significant increase in [Ca(2+)]i and [Na(+)]i in myocytes when compared to controls, which was significantly reduced by halothane in a dose-dependent fashion. In the presence of TTX and NIF, halothane also significantly reduced the rise in the [Ca(2+)]i and [Na(+)]i in myocytes subjected to MI. 1-heptanol, another gap junction blocker, had similar effects. Thus, halothane reduced [Ca(2+)]i and [Na(+)]i overload produced by MI in myocytes. This effect is not solely due to block of voltage-gated Na(+) and Ca(2+) channels, and is likely mediated by inhibiting the opening of connexin-43 hemichannels. PMID- 11735262 TI - p38 MAPK activation triggers pharmacologically-induced beta-adrenergic preconditioning, but not ischaemic preconditioning. AB - p38 Mitogen-activated protein kinase (p38 MAPK) is activated by short episodes of ischaemia-reperfusion as well as by sustained ischemia followed by reperfusion, Whether activation of this kinase is beneficial or deleterious to the ischaemic heart is still a subject of controversy. Since transient beta-adrenergic stimulation (5 min) stimulates p38 MAPK activation and mimics the cardioprotection of ischaemic preconditioning, it was used as a tool to further evaluate the role of this kinase in cardioprotection. The isolated perfused working rat heart, subjected to 25 min ischaemia and 30 min reperfusion was used as experimental model. p38 MAPK and ATF2 activation was determined using Western blots. The results showed that isoproterenol stimulated p38 MAPK in a dose- and time-dependent manner. Ischaemia-induced activation of p38 MAPK could be partially abolished by beta- and alpha1-adrenergic receptor blockade. Isoproterenol activation of the kinase could be abolished by alprenolol and verapamil, but not by 8-cyclopentyladenosine. p38 MAPK activation induced by either a multi-episode preconditioning protocol or isoproterenol (10(-7) M for 5 min) was associated with a significant reduction in p38 MAPK activation at all time intervals studied during 25 min global ischaemia and at 20 and 30 min of reperfusion, compared with the marked activation observed in untreated non preconditioned hearts. In each case attenuation of p38 MAPK activation during ischaemia and during reperfusion was associated with improved functional recovery during reperfusion. Cyclic elevations in tissue cAMP during an ischaemic preconditioning protocol acted as trigger of cardioprotection, since pretreatment of such hearts with alprenolol abolished cardioprotection. Mechanical failure in such hearts was characterized by a significant stimulation of p38 MAPK activity during ischaemia and reperfusion. However, p38 MAPK activation during an ischaemic preconditioning protocol did not act as trigger: inhibition of p38 MAPK activation by SB 203580 during the preconditioning phase did not abolish cardioprotection. In fact, functional recovery was significantly better than that of untreated preconditioned hearts. On the other hand, SB 203580, when administered before and during the isoproterenol-preconditioning protocol abolished cardioprotection, suggesting that p38 MAPK activation by a beta adrenergic-induced preconditioning protocol does act as trigger of cardioprotection. In addition, attenuation of p38 MAPK activity during sustained ischaemia and reperfusion as occurs in ischaemic- or isoproterenol-preconditioned hearts, is beneficial. PMID- 11735263 TI - Morphine sulfate inhibits hypoxia-induced vascular endothelial growth factor expression in endothelial cells and cardiac myocytes. AB - Vascular endothelial growth factor (VEGF) is an angiogenic mitogen, specific for endothelial cells. Hypoxia-induced VEGF in endothelial cells and cardiomyocytes leads to autocrine and paracrine stimulation, respectively. During myocardial ischemia, VEGF is upregulated in the endothelium and myocardium, and may mediate angiogenesis. Morphine sulfate is commonly used in pain relief for patients with acute myocardial infarction. We investigated the effect of morphine sulfate on VEGF expression in cultured endothelial cells and cardiac myocytes subjected to hypoxia. Enzyme-linked immunosorbent assays showed that morphine sulfate significantly inhibited hypoxia-induced VEGF expression in mouse heart microvascular endothelial cells (SMHEC4), primary cultures of human umbilical vein endothelial cells (HUVECs) and in primary cultures of rat cardiac myocytes (P<0.05). Real time reverse transcriptase polymerase chain reaction showed that morphine treatment (100 ng/ml) of hypoxic HUVECs resulted in a significant reduction in mRNA levels of VEGF(121) and VEGF(165) isoforms. Transfection of HUVECs with a human VEGF promoter-luciferase construct showed that hypoxia induced transcriptional activation of VEGF was markedly inhibited by morphine sulfate (P<0.05). Phosphatidyl inositol-3 kinase and protein kinase C-mediated activation of the VEGF promoter was also inhibited by morphine. The opioid antagonist naloxone significantly reversed the inhibitory effects of morphine in endothelial cells suggesting the involvement of opioid receptors. Our results show that the inhibitory effects of morphine on hypoxia-induced VEGF expression in endothelial cells and cardiac myocytes can lead to a decrease in the autocrine and paracrine stimulation and hence limit neovascularization of the ischemic myocardium. PMID- 11735264 TI - Taurolidine improves survival by abrogating the accelerated development and proliferation of solid tumors and development of organ metastases from circulating tumor cells released following surgery. AB - BACKGROUND: Surgical trauma is partly responsible for enhancing tumor growth through a variety of mechanisms that are still not fully elucidated. The use of perioperative immunostimulants may modulate these effects. This study examined the effect of administration of taurolidine after laparotomy or laparoscopy on the growth of solid tumor, the establishment of hepatic and lung metastases, and effects on natural killer (NK) and lymphokine-activated killer (LAK) cell function. METHODS: B16 melanoma right flank tumors were established in mice (n = 180). These animals underwent anesthesia only (control) or laparotomy or laparoscopy (n = 60 per group) and were randomized to receive either saline or taurolidine (n = 30 per group) at specific time points. Survival was determined in each group, and in a further 90 mice tumor growth was followed over 10 days postoperatively. The experiment was repeated in 540 mice, which underwent one of the three procedures and were treated with either saline or taurolidine. NK and LAK cell cytotoxicity (NKCC, LAKCC) was determined at several time points postoperatively. In a further experiment, B16 melanoma tumor cells were delivered via tail vein injection (n = 180) and intrasplenic injection (n = 180). The effect of saline or taurolidine administration on survival after the establishment of metastases was determined, and again in a further 180 mice the establishment of metastatic deposits in the liver or lungs was determined after 8 days. RESULTS: Survival appeared to be significantly decreased in both the solid tumor model and the metastatic models undergoing laparotomy compared to laparoscopy and controls (P < 0.0001) and to a lesser extent in the laparoscopy group compared to controls (P < 0.001). Flank tumor growth and metastatic tumor formation were more significant in laparotomy groups compared to laparoscopy groups and controls, but also to a lesser extent in laparoscopy groups compared to controls (P 0.05), and the PDE III inhibitor milrinone had a comparable effect. Moreover, dbcAMP also attenuated TNF-alpha production with abrogation of the synergistic effect of hypoxia (0.56 +/- 0.08 ng/10(6) cells in dbcAMP plus hypoxia plus LPS vs 0.46 +/- 0.04 ng/10(6) cells in dbcAMP plus LPS, P > 0.05). CONCLUSIONS: The results show that elevation of cellular cAMP, either by promotion of generation or by inhibition of degradation, suppresses LPS-stimulated TNF-alpha production in hypoxic M phi. It appears that hypoxia synergizes with LPS in the induction of M phi TNF-alpha production through PDE-mediated cAMP degradation. Inhibition of PDE may be a therapeutic approach for suppression of synergistic induction of M phi TNF-alpha production by hypoxia and LPS in posttraumatic tissue. PMID- 11735280 TI - TNF receptor I mediates chemokine production and neutrophil accumulation in the lung following systemic lipopolysaccharide. AB - BACKGROUND: Tumor necrosis factor (TNF)-alpha is a critical effector of lipopolysaccharide (LPS)-induced acute lung injury, and its effects are mediated by two structurally related receptors, RI and RII. Cellular adhesion molecules and C-X-C chemokines (Keratinocyte chemoattractant (KC) and macrophage inflammatory protein [MIP]-2) regulate tissue neutrophil polymorphonuclear neutrophil (PMN) accumulation in a multitude of inflammatory states. We hypothesized that TNFRI signaling dictates PMN accumulation in the lung via regulation of chemokine molecule production. Therefore, the purposes of this study were to (1) delineate LPS-induced lung TNF-alpha production and (2) characterize the contribution of both TNF receptors to lung chemokine production and neutrophil influx following systemic LPS. METHODS: Wild-type or TNFRI and TNFRII knockout (KO) mice were injected with vehicle (saline) or LPS (Escherichia coli 0.5 mg/kg intraperitoneally). After 2, 4, 6, or 24 h, lungs were analyzed for TNF-alpha and chemokine (KC and MIP-2) protein expression (enzyme-linked immunosorbent assay) and PMN accumulation (myeloperoxidase assay). RESULTS: There was an increase in total lung TNF-alpha (vehicle, 5.0 +/- 1.2 pg/mg total protein vs LPS, 950 +/- 318; P < 0.05) after LPS. Lung chemokine production and PMN accumulation were also increased compared to vehicle-injected mice. Lung chemokine production and PMN accumulation were significantly lower in TNFRI KO, but not TNFRII KO, mice, despite no difference in TNF-alpha production (TNFRI KO, 925 +/- 301 vs TNFRII KO, 837 +/- 267, P = 0.82). CONCLUSIONS: Acute lung injury following systemic LPS administration is characterized by increased lung (1) TNF alpha production, (2) C-X-C chemokine production, and (3) neutrophil accumulation. The maximal effect of LPS-induced lung neutrophil accumulation appears to be dependent upon the TNFRI receptor but not the TNFRII receptor. . PMID- 11735281 TI - Effect of perfusion during ischemia on skeletal muscle. AB - BACKGROUND: Despite the established preservation method for major organs of perfusion followed by immersion at hypothermia, a standard preservation technique for skeletal muscle is still a matter of controversy. The purpose of this study is to examine the effect of perfusion on the preservation of skeletal muscle in amputated limbs. MATERIALS AND METHOD: The rat hindlimb was amputated for perfusion with Euro-Collins (EC) or University of Wisconsin (UW) solution at different perfusion pressures (40 or 100 cm-gravity). After certain ischemic periods (4 or 5 h), the skeletal muscle viability was determined by measuring the tissue content of adenosine triphosphate (ATP). RESULT: The UW solution perfusion group maintained better ATP levels than the EC solution group when the ischemic period was extended to 5 h. The perfusion pressure of 100 cm-gravity was more effective for preserving muscle viability than 40 cm-gravity with both EC and UW solutions. CONCLUSION: UW solution might be adequate to preserve muscle viability and perfusion pressure is recommended at 100 cm-gravity rather than at minimal pressure (40 cm-gravity), which washes out stagnant blood. PMID- 11735282 TI - Clinical significance of matrix metalloproteinase-7 and Ets-1 gene expression in patients with lung cancer. AB - BACKGROUND: Matrix metalloproteinase-7 (MMP-7) is a member of the MMP family and has a wide variety of substrate spectra. Ets domain transcription factors are reported to play an important role in carcinoma invasion and metastasis. The regulatory role of Ets-1 has been shown in several MMPs. We have hypothesized that MMP-7 and Ets-1 mRNA levels could be predictors of the development and invasion of lung cancer. METHODS: The study included 73 lung cancer cases. The mRNA levels were quantified by real-time reverse transcription-polymerase chain reaction (RT-PCR) using a LightCycler. RESULTS: No significant difference in MMP 7 and Ets-1 mRNA levels was found among gender, age, and pathological subtype. The MMP-7 mRNA levels were elevated in tumor tissues from stage II-IV lung cancer (1.629 +/- 2.267) compared to those from stage I lung cancer (0.762 +/- 1.463) (P = 0.0290). There was a tendency toward higher MMP-7 mRNA expression levels in tumors with lymph node metastasis (1.728 +/- 2.432) compared to those without lymph node metastasis (1.141 +/- 1.838) (P = 0.1076). Thus, MMP-7 mRNA levels may serve as a marker of higher stages in lung cancer. No significant difference in Ets-1 mRNA levels was found among clinical stages and T-status. The Ets-1 mRNA levels were elevated in tumors from N2 patients (7.512 +/- 13.306) compared to those from N0 patients (2.525 +/- 4.719) (P = 0.0209). Ets-1 mRNA levels showed a positive correlation with MMP-7 expression (P = 0.0042). CONCLUSIONS: Using the LightCycler RT-PCR assay, the determination of MMP-7 and Ets-1 mRNA levels might provide a potential marker for advanced lung cancer. However, further studies and a longer follow-up are needed to confirm the impact of MMP-7 in the biological behavior of the tumor. PMID- 11735283 TI - Polyanionic polysaccharides reduce intra-abdominal adhesion and abscess formation in a rat peritonitis model. AB - BACKGROUND: Intra-abdominal infection is complicated by adhesion and abscess formation. We have assessed the adhesion- and abscess-reducing capacity of various solution volumes and concentrations of two polyanionic polysaccharides, hyaluronan (HA) and carboxymethylcellulose (CMC), in a rat peritonitis model. STUDY DESIGN: In 192 male Wistar rats a bacterial peritonitis was induced using cecal ligation and puncture. After 24 h the abdomen was reopened and the ligated cecum resected. Animals were randomized into three control groups, nine groups treated with various solution volumes (1 to 8 ml) containing different HA concentrations, and four groups treated with 1.7% CMC solution. Rats were killed at day 7, postoperatively, and adhesions were scored at five abdominal sites on a scale from 0 to 4. The presence and size of intra-abdominal abscesses were noted. RESULTS: Fifty-four rats (28%) prematurely died. There was no significant difference in mortality between treatment groups and controls. Treatment with CMC (P < 0.001) and low (0.2 and 0.4%) concentrations of HA (P < 0.005) significantly reduced intra-abdominal adhesion formation. High volumes of 0.2 and 0.4% HA were most effective (P = 0.01). The effect of CMC was volume independent. The incidence of abdominal abscesses was also significantly reduced by treatment with either CMC (P < 0.001) or low concentrations of HA (P < 0.001). With regard to abscess formation the effect was independent of the volume administered for HA, while low volumes of CMC were most effective (P < 0.005). CONCLUSION: Intraperitoneal treatment with either CMC or low-viscosity HA solution reduced intra-abdominal adhesion and abscess formation in a rat peritonitis model. The volume-induced reduction in adhesion formation suggests a hydroflotation effect of HA solution. PMID- 11735284 TI - A cellular automaton model of early tumor growth and invasion. AB - A hybrid cellular automaton model is described and used to simulate early tumor growth and examine the roles of host tissue vascular density and tumor metabolism in the ability of a small number of monoclonal transformed cells to develop into an invasive tumor. The model incorporates normal cells, tumor cells, necrotic or empty space, and a random network of native microvessels as components of a cellular automaton state vector. Diffusion of glucose and H(+)ions (the latter largely resulting from the tumor's excessive reliance on anaerobic metabolism) to and from the microvessels, and their utilization or production by cells, is modeled through the solution of differential equations. In this way, the cells and microvessels affect the extracellular concentrations of glucose and H(+)which, in turn, affect the evolution of the automaton. Simulations of the model demonstrate that: (i) high tumor H(+)ion production is favorable for tumor growth and invasion; however for every H(+)ion production rate, there exists a range of optimal microvessel densities (leading to a local pH favorable to tumor but not to normal cells) for which growth and invasion is most effective, (ii) at vascular densities below this range, both tumor and normal cells die due to excessively low pH, (iii) for vascular densities above the optimal range the microvessel network is highly efficient at removing acid and therefore the tumor cells lose their advantage over normal cells gained by high local H(+)concentration. While significant spatial gradients of glucose formed, no regions of detrimentally poor glucose perfusion (for either cell type) were observed, regardless of microvessel density. Depending on metabolic phenotype, a variety of tumor morphologies similar to those clinically observed were realized in the simulations. Lastly, a sharp transition (analogous to that of the adenoma carcinoma sequence) between states of initial tumor confinement and efficient invasiveness was observed when H(+)production reached a critical value. PMID- 11735285 TI - Tripedal knuckle-walking: a proposal for the evolution of human locomotion and handedness. AB - A comparative morphological analysis of human and non-human hominoids was conducted in an attempt to determine the mode of locomotion of the protohominid. Although the generalized hominoid anatomy permits variation of locomotion: brachiation, knuckle-walking, etc., minor variations in structure determine which behavior is favored. Arboreal arm swinging requires a flexible forelimb while terrestrial fist or knuckle-walking demands more rigidity of the hand and wrist. It is demonstrated that the large human thumb accompanied by the strong adduction of the thenar, hypothenar, and palmar interosseous muscles offer powerful rigidity to the hand, while fusion of the os centrale with the scaphoid during gestation permits the formation of an arch of carpals which imbue the wrist with the stability necessary for weight bearing. Fascialization of the contrahentes and dorsiepitrochlearis muscles in the human as well as depilation of the middle phalanges; the webbing (syndactyly) of the palm; the direction of the fibers of the interosseous membrane of the forearm; the shape of the puerile annular ligament, and the direction of the human glenoid fossa strongly suggest that the ancestor of man used a knuckle-walking form of locomotion prior to becoming bipedal. A model is presented that suggests that bipedalism was attained through an intermediate stage of tripedalism. The model is based on the fact that man's anatomy is much more asymmetric than that of the great apes. A presumption is made that due to the absence of trees for climbing in the transition from forest to open plain, the protohominid needed to carry tools (stones) at all times for protection. Stones could be carried for long distances on the posterior iliac crest since the weight would be shifted posteriorly over the legs. Pick up, medial rotation and adduction of the stone would employ a two-muscle chain of biceps brachii and latissimus dorsi. On the iliac crest, the stone is posterior to the coronal plane of the glenohumeral joint, and with the contraction of this two-muscle chain, the shoulder on one side is moved posteriorly effecting a semi erect posture. It is proposed that tripedalism of the protohominid may be an explanation for the handedness unique to hominids. PMID- 11735286 TI - A physical origin for functional domain structure in nucleic acids as evidenced by cross-linking entropy: I. AB - A global strategy for estimating the entropy of long sequences of RNA is proposed to help improve the predictive capacity of RNA secondary structure dynamic programming algorithm (DPA) free energy (FE) minimization methods. These DPA strategies only consider the effects that occur in the immediate (nearest neighbor) vicinity of a given base pair (bp) in a secondary structure plot. They are therefore defined as nearest-neighbor secondary structure (NNSS) strategies. The new approach utilizes the statistical properties of the Gaussian polymer chain model to introduce both local and global contributions to the entropy of a given secondary structure. These entropic contributions are primarily a function of the persistence length of the RNA. Limits on the domain size are strongly suggested by this model and these limits are a function of both the length and the percentage of bp enclosed within a given domain. The model generalizes the penalties found in the NNSS algorithms. The approach considers the importance of flexibility in the folding and stability of RNA by considering the role of the persistence length in a biopolymer structure. The theory also suggests that molecular machinery may also take advantage of this global entropic effect to bring about catalytic effects. The applications can also be extended to protein structure calculations with some additional considerations. PMID- 11735287 TI - A physical origin for functional domain structure in nucleic acids as evidenced by cross-linking entropy: II. AB - In Part I, cross-linking entropy (CLE) was proposed as a mechanism that limits the size of functional domains of RNA. To test this hypothesis, the theory is developed into an RNA secondary structure prediction filter which is applied to nearest-neighbor secondary structure (NNSS) algorithms that utilize a free energy (FE) minimization strategy. (The NNSS strategies are also referred to as the dynamic programming algorithm in the literature.) The cross-linking entropy for RNA is derived from a generalized Gaussian polymer chain model where the entropic contributions caused by the formation of base pairs (stacking) in RNA are analysed globally. Local entropic contributions are associated with the freezing out of degrees of freedom in the links. Both global and local entropic effects are strongly influenced by the persistence length. The cross-linking entropy provides a physical origin for the size of functional domains in long nucleic acid sequences and may go further to explain as to why the majority of the domain regions in typical sequences tend to be less than 600 nucleotides in length. In addition, improvements were observed in the "best guess" predictive capacity over NNSS prediction strategies. The thermodynamic distribution is more representative of the expected structures and is strongly governed by such physical parameters as the persistence length and the excluded volume. The CLE appears to generalize the tabulated penalties used in NNSS algorithms. The principal parameter influencing this entropy is the persistence length. The model is shown to accomodate a variable persistence length and is capable of describing the folding dynamics of RNA. A two-state kinetic model based on the CLE principle is used to help elucidate the folding kinetics of functional domains in the group I introns. PMID- 11735288 TI - The use of mixed distribution models to determine bout criteria for analysis of animal behaviour. AB - Feeding behaviour consists of feeding events, separated by non-feeding intervals. Feeding events are often clustered into bouts, which may be called meals. Grouping feeding events into meals requires the determination of a bout or meal criterion, that is, the longest interval accepted as part of a meal. Tolkamp & Kyriazakis (1999a) proposed a three-Gaussian model to estimate meal criteria. The three Gaussians each described the frequency distribution of the log(e) transformed lengths of a population of intervals. These populations were thought to be: (1) short intervals within meals; (2) intervals within meals during which animals drink; (3) intervals between meals. This model predicted that the probability of an animal starting a meal would first increase, and then decrease with time since the last meal. This contrasts with expectations based on the satiety concept, which predicts that the probability of an animal starting a meal will increase with time since the last meal. This discrepancy is related to the symmetrical nature of the Gaussian distribution. Alternatively, the two-parameter Weibull distribution can take a skewed form and perhaps is more suitable to describe the different populations of intervals. In this study, models consisting of combinations of Gaussian and Weibull distributions were examined for their suitability to describe the observed feeding behaviour of cows. Weibulls did not improve the description of the populations of within-meal intervals, compared to Gaussians. However, the Weibull distribution was found to describe the between meal population of intervals statistically better than the Gaussian. Additionally, this inclusion of a Weibull, as opposed to a Gaussian, resulted in predictions that were in better agreement with the satiety concept over the entire range of interval lengths observed. A model based on Gaussians to describe the within-meal populations of intervals and a Weibull to describe the population of between-meal intervals is, therefore, proposed. This model leads to biologically more satisfactory estimates of bout criteria than previous models and is likely to be applicable both across species and behaviours. PMID- 11735289 TI - Data estimation and the colour of time series. AB - There has been a long debate on the source of temporal fluctuations in natural population densities. The difficulty is that unpredictable irregularities might be attributed either to external environmental factors or to chaotic dynamics of populations, or even to the interaction of these two factors. Some years ago Cohen (1995) pointed out that real time series follow redshifted Fourier power spectra, while the simplest chaotic population dynamical models are mostly blueshifted. Since then, the controversy has focused on comparisons of Fourier spectra originating from different models and data. Here, we show experimentally that estimation process by human observers shifts power spectra to the red. This result implies that because of estimation distortion, real population data must be less redshifted than many recorded time series suggest. PMID- 11735290 TI - Theoretical analysis of systematic errors introduced by a pedicel-girdling technique used to estimate separately the xylem and phloem flows. AB - The water budget of fruits was analysed by means of a biophysical model of fruit growth, built and calibrated recently for peaches [Prunus persica (L.) Batsch]. This analysis was applied to the evaluation of systematic errors introduced by a pedicel-girdling method (with the observations being treated by means of a subtractive technique) used to separate the contributions of xylem and phloem flow to the total water inflow to the fruit. The flows were considered as solution transport through composite membranes and were calculated by means of equations drawn from non-equilibrium thermodynamics. The total inflow of water was simulated as dependent on the water status in the tree. The hourly time step was used for the simulation. The flows obtained by simulation of the pedicel girdled fruit were compared with those found by simulation of the intact-pedicel fruit. The error introduced by the pedicel-girdling technique was evaluated theoretically and was shown to vary during the day, ranging from very small (relative error of 3-7%) at the period when the rate of fruit growth is maximal to 100% when the fruit volume does not change. The vascular flows obtained from the "girdling experiments" are discussed in relation to the possible theoretically estimated errors. PMID- 11735291 TI - Helper-dependent vs. helper-independent CTL responses in HIV infection: implications for drug therapy and resistance. AB - Clinical data from HIV-infected patients, as well as theoretical studies, suggest that CTL responses in the presence and absence of CD4 cell help are qualitatively different. In the presence of help, CTL responses are maintained despite very low antigenic loads and control the infection in the long term. In the absence of specific helper cell responses, CTL require high antigenic loads to be maintained, are short lived at low levels of antigen, and do not control the infection in the long term. This paper describes mathematical models analysing the dynamics of helper-dependent and helper-independent CTL in HIV infection with special focus on the dynamics during drug therapy in chronic infection. Theory suggests that a fast rate of virus spread results in high degrees of helper cell impairment which promotes the development of helper-independent CTL responses and compromised immunological control. In agreement with clinical findings, the model suggests that upon start of therapy, there is a transient increase in the level of CTL, followed by a decline to low levels once virus load has been significantly suppressed. According to the model, the presence of helper independent CTL can promote the establishment of a helper-dependent memory response. Interestingly, this gives rise to the prediction that a relatively early stop of therapy, before the level of CTL has fallen below a threshold, can promote improved immunological control. Issues concerning the timing and duration of treatment are discussed. The CTL kinetics during drug therapy also provide new insights into the principles underlying the emergence of drug-resistant strains during the course of treatment. PMID- 11735292 TI - Social inhibition and the regulation of temporal polyethism in honey bees. AB - Honey bee division of labor is characterized by temporal polyethism, in which young workers remain in the hive and perform tasks there, whereas old workers perform more risky outside tasks, mainly foraging. We present a model of honey bee division of labor based on (1) an intrinsic process of behavioral development and (2) inhibition of development through social interactions among the workers in a colony. The model shows that these two processes can explain the main features of honey bee temporal polyethism: the correlation between age and task performance; the age at which a worker first forages and how this age varies among hives; the balanced allocation of workers to hive tasks and foraging; the recovery of a colony from demographic perturbations; and the differentiation of workers into different behavioral roles. The model provides a baseline picture of individual and colony behavior that can serve as the basis for studies of more fine-grained regulation of division of labor. PMID- 11735293 TI - The molecular roots of compositional inheritance. AB - Non-covalent compositional assemblies, made of monomeric mutually catalytic molecules, constitute an alternative to alphabet-based informational biopolymers as a mechanism of primordial inheritance. Such assemblies appear implicitly in many "Metabolism First" origin of life scenarios, and more explicitly in the Graded Autocatalysis Replication Domain (GARD) model [Segreet al. (2000). Proc. Natl Acad. Sci. U.S.A.97, 4112-4117]. In the present work, we provide a detailed analysis of the quantitative molecular roots of such behavior. It is demonstrated that the fidelity of reproduction provided by a newly defined heritability measure eta(*)(s), strongly depends on the values of molecular recognition parameters and on assembly size. We find that if the catalytic rate acceleration coefficients are distributed normally, transfer of compositional information becomes impossible, due to frequent "compositional error catastrophes". In contrast, if the catalytic acceleration rates obey a lognormal distribution, as actually predicted by a statistical formalism for molecular repertoires, high reproduction fidelity is obtained. There is also a clear dependence on assembly size N, whereby maximal eta is seen in a narrow range around N approximately 3.5 N(G)/lambda, where N(G)is the size of the primordial molecular repertoire and lambda is a molecular interaction statistical parameter. Such relationships help define the physicochemical conditions that could underlie the early steps in pre biotic evolution. PMID- 11735294 TI - The prediction of the structural class of protein: application of the measure of diversity. AB - Based on the concept that the structural class of a protein is mainly determined by its secondary structure sequence, a new algorithm for prediction of the structural class of a protein is proposed. By use of the number of alpha helices, beta -strands, and betaalphabeta fragments, the structural class of a protein can be predicted by an algorithm based on the increment of diversity (ID), in which the sole prediction parameter-the increment of diversity is used as the index of prediction of structural class of a protein. The results indicate that the high rates of correct prediction are obtained for complete set (standard set) from Brookhaven Protein Data Bank-CD ROM (PDB) published in October 1995 and the test set newly released from Brookhaven Protein Data Bank-CD ROM (PDB) before July 1998, respectively. PMID- 11735295 TI - A modified mutagenic PCR-RFLP method for K-ras codon 12 and 13 mutations detection in NSCLC patients. AB - Evidence from many investigators has shown that mutations in the first exon of K- ras gene occur at elevated frequencies in lung, pancreatic and colon carcinoma and seem to be of prognostic importance. The aim of this study was to develop an effective method for the detection of K- ras mutations in codons 12 and 13 in non small-cell lung cancer (NSCLC) patients in order to investigate correlation with clinical outcome. DNA was extracted from tumour and neighbouring non-neoplastic lung tissues from 70 patients and screened for codon 12 and 13 mutations. We applied a mutagenic PCR-restriction fragment length polymorphism for both codon 12 and 13 mutation detection. Codon 12 mutation was identified in 20% of NSCLC patients, whereas no codon 13 mutation was detected. As expected, the respective non-neoplastic tissues exhibited no mutations. We observed an increased codon 12 mutation prevalence in adenocarcinoma comparing to other types of carcinomas. Follow-up for 29 patients with a mean time of 12 months indicates an increased relapse rate in NSCLC patients with the K- ras codon 12 mutation. Furthermore, a trend towards increased percentage of mutant samples was observed in the advanced stage group of patients. We provide evidence that our approach is a fast and reliable method for screening K- ras exon 1 mutations in tumour samples from NSCLC patients. PMID- 11735296 TI - Identification of Staphylococcus aureus based on PCR amplification of species specific genomic 826 bp sequence derived from a common 44-kb Sma I restriction fragment. AB - Primers were designed for polymerase chain reaction (PCR)-amplification of a genomic sequence specific to Staphylococcus aureus strains. The sequence corresponds to a part of the 44-kb Sma I fragment (fragment L on the S. aureus NCTC 8325 restriction map) which was found to be common to strains of the S. aureus species (Pantucek et al 1996, International Journal of Systematic Bacteriology, 46: 216-222). The labelled 44-kb Sma I restriction fragment derived from S. aureus NCTC 8325-4 was hybridized to the Eco RI restriction patterns of genomic DNA from 13 strains representing different macrorestriction types of S. aureus subsp. aureus. This made it possible to reveal the 2052 bp Eco RI restriction subfragment and to demonstrate its presence in all the tested strains. From the sequence of this subfragment, primers were designed by means of which the 826 bp amplicons were obtained in all 216 tested strains of S. aureus. No hybridization and PCR-products were observed in 40 collection strains of other staphylococcal species and subspecies as well as in 45 clinical strains of coagulase-negative staphylococci. These results lead us to the conclusion that the use of the above primers makes it possible to identify rapidly and reliably S. aureus strains of various provenance and different genotypes. PMID- 11735297 TI - Development and evaluation of a fluorogenic 5'-nuclease assay to identify Marburg virus. AB - The ability to rapidly recognize Marburg virus infections is critical to quickly institute proper barrier nursing precautions and limit further spread of the disease. A rapid, sensitive, and specific laboratory diagnostic test is necessary to confirm outbreaks of Marburg virus and to distinguish it from other diseases that can present with similar clinical symptoms. A one-tube reverse transcriptase polymerase chain reaction (RT-PCR) assay for the identification of Marburg virus was developed and evaluated using the ABI PRISM 7700 Sequence Detection System and TaqMan chemistry. The sensitivity and specificity of the newly designed primer/probe set (MBGGP3) was evaluated. MBGGP3 was equivalent to or 10-100-fold more sensitive than previously designed primer sets as determined by limit of detection experiments. In addition, the MBGGP3 assay was able to detect all strains of Marburg virus tested, but gave negative results with other haemorrhagic fever and genetically related viruses. The results of this study indicate that the MBGGP3 primer/probe set is both sensitive and specific. In addition, this assay is compatible with emerging rapid nucleic acid analysis platforms and therefore may prove to be a useful diagnostic tool for the control and management of future outbreaks. PMID- 11735298 TI - Sample preparation methods for PCR detection of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on beef chuck shoulder using a single enrichment medium. AB - To improve the utility of the polymerase chain reaction (PCR) for food samples, methods for preparing template DNA were developed to remove PCR inhibitors. Beef chuck shoulder medallions, artificially contaminated, individually or in combination, with Escherichia coli serotype O157:H7 strain FSIS 45753-35, Salmonella typhimurium DT104 strain 13HP, or Listeria monocytogenes strain Scott A at concentrations of 10, 1 and 0.5 cfu/cm(2)were swabbed with a sponge, and the sponges were enriched for 18 h at 37 degrees C in universal pre-enrichment broth (UPB). Enriched broth cultures (EBC), cell pellets (CP), or phosphate-buffered saline-washed cell pellets (PBSCP) from enriched sponge samples were compared for detection of E. coli O157:H7, S. typhimurium DT104, or L. monocytogenes by the PCR using the BAX(TM)system. Recovery of the three organisms was effective for detection of each pathogen at initial levels of 10, 1 and 0.5 cfu/cm(2)when inoculated separately, or in combination, onto the beef samples. Use of EBC, CP, or PBSCP of sponge-swabbed samples eliminated problems associated with inhibition of the PCR by food components, time-consuming extraction of DNA, and inhibition due to large amounts of non-target DNA derived from the food. The procedure involving enrichment of sponge-swabbed beef samples in UPB followed by PCR amplification using EBC with the BAX system is the most efficient and simple method for detection of E. coli O157:H7, S. typhimurium DT104, and L. monocytogenes. PMID- 11735299 TI - Rapid detection of Listeria monocytogenes in foods, by a combination of PCR and DNA probe. AB - Listeria monocytogenes is a frequent contaminant of water and foods. Its rapid detection is needed before some foods can be prepared for marketing. In this work L. monocytogenes has been searched for in foods, by a combination of polymerase chain reaction (PCR) and a DNA probe. Both PCR and the probe were prepared for recognizing a specific region of the internalin gene, which is responsible for the production of one of the most important pathogenic factors of Listeria. The combined use of PCR and the DNA probe was used for the detection of L. monocytogenes in over 180 environmental and food samples. Several detection methods were compared in this study, namely conventional culture methods; direct PCR; PCR after an enrichment step; a DNA probe alone; a DNA probe after enrichment and another commercially available gene-probe. Finally PCR and the DNA probe were used in series on all the samples collected. When the DNA probe was associated with the PCR, specific and accurate detection of listeria in the samples could be obtained in about a working-day. The present molecular method showed some advantages in terms of rapidity and specificity in comparison to the other aforementioned tests. In addition, it resulted as being easy to handle, even for non-specialized personnel in small diagnostic microbiology laboratories. PMID- 11735300 TI - Mutation screening for Japanese Lafora's disease patients: identification of novel sequence variants in the coding and upstream regulatory regions of EPM2A gene. AB - The progressive myoclonus epilepsy of Lafora type (LD) is an autosomal recessive disorder caused by mutations in the EPM2A gene. We demonstrated recently that EPM2A encodes a dual-specificity phosphatase that is primarily associated with polyribosomes. In the present study, we screened for mutations in the EPM2A gene in 4 Japanese LD families and identified a novel mis-sense mutation, Ala46Pro (136G-->C), in heterozygous condition in one patient. In addition, sequence analyses in the patient and control DNA samples identified 4 single nucleotide polymorphisms (SNPs) (75G/A, 120G/T, 159C/G, 171C/T) in the coding region and a novel insertion/deletion polymorphic site (-483[T](11/10)[A](2/3)) and a SNP ( 547A/G) in the putative regulatory region of the EPM2A gene. None of the sequence variants, however, co-segregated with the LD phenotype. Haplotype analysis for the 6q24 region in the affected families revealed lack of homozygosity at the EPM2A locus. Our studies suggest that EPM2A is not involved in the disease phenotype of the 4 families studied and that locus heterogeneity for LD may exist in Japanese population also. A simple test described for the detection of Ala46Pro mutation present heterozygously in Japanese population (allele frequency 0.026) can be used for screening this novel allele in a larger sample size. PMID- 11735301 TI - Development of a PCR ELISA assay for the identification of Campylobacter jejuni and Campylobacter coli. AB - A polymerase chain reaction (PCR) assay was developed based on a solution hybridization colorimetric end-point detection format (PCR ELISA) for the identification of Campylobacter jejuni and Campylobacter coli. PCR primers were designed to target a gene sequence with species-specific motifs. Five biotin labelled probes targeted to the species-specific motifs were investigated for the detection of digoxygenin-labelled PCR products from C. jejuni and C. coli using the PCR ELISA format. Two probes were identified, one which reacts with both the C. jejuni and C. coli target sequences (probe CC2) and one probe which reacts with the C. jejuni target sequence only (probe CJ2). The specificity of the assay with the CJ2 and CC2 probes was investigated with a range of Campylobacter spp., Arcobacter spp., Helicobacter spp. and a range of unrelated organisms. The PCR ELISA assay and probes were demonstrated to be specific for C. jejuni and C. coli. The sensitivity of the PCR ELISA assay was demonstrated to be 10-100-fold more sensitive than a gel-based PCR method using the same primers. This PCR ELISA assay is sensitive, specific and significantly reduces the time needed for the identification of C. jejuni and C. coli and has the potential to facilitate early detection of these important gastro-intestinal pathogens. PMID- 11735302 TI - Electroporation of DNA sequences from the pathogenicity locus (PaLoc) of toxigenic Clostridium difficile into a non-toxigenic strain. AB - Toxigenic Clostridium difficile is the etiologic agent of C. difficile-associated diarrhoea (CDAD), the most common cause of hospital-acquired infectious diarrhoea. The genes tcdA and tcdB, which encode for the toxin A and B proteins, are part of the pathogenicity locus (PaLoc) of toxigenic C. difficile. Genetic and virulence studies at the molecular level in C. difficile have been hindered by the lack of techniques for DNA manipulation in this species. We describe the electroporation of DNA fragments from a toxigenic isolate into a non-toxigenic strain of C. difficile. Using previously described methods of electroporation into Clostridium spp., the complete toxin B gene and polymerase chain reaction (PCR) fragments of the PaLoc were cloned and electroporated into a non-toxigenic strain of C. difficile. The resulting transformed clones were screened for the introduced gene fragments by PCR, which confirmed their presence. This is the first description of introduction of DNA into C. difficile by electroporation. PMID- 11735303 TI - Beta-actin--an unsuitable internal control for RT-PCR. AB - Despite reports confirming cell-cycle dependent gene expression and a number of studies describing specific circumstances in which beta-actin is also regulated, the mRNA for beta-actin remains a widely used housekeeping gene internal control. Utilizing differential reverse transcriptase-polymerase chain reaction (RT-PCR), we report here the dose-dependent inhibition of beta-actin by matrigel. This was detected by comparison to the very moderate inhibition of the target gene, membrane type-1 matrix metalloproteinase (MT1-MMP), with results independently confirmed by similar findings on MT1-MMP expression using competitive RT-PCR. Furthermore, RT-PCR of the housekeeping gene 18 Svedberg Units (S) rRNA demonstrated excellent consistency, reproducibility and non-regulation by a matrigel treatment. We conclude that beta-actin is highly regulated by matrigel and therefore unsuitable as an internal control in this treatment. Hence, these findings suggest that researchers have a responsibility to ensure that the housekeeping gene of choice is not regulated in their specific application, as such regulation may dramatically affect the accuracy of their results. This study reinforces the necessity for minimally regulated housekeeping genes such as 18S rRNA, and the superiority of competitive templates as internal controls for quantitative applications of RT-PCR. PMID- 11735304 TI - Characterization of microsatellite markers adjacent to AP-4 on chromosome 16p13.3. AB - The 1400 kb genomic sequence between the markers D16S406 and D16S423 on chromosome 16p13.3 has been recently sequenced and the interval contains a transcription factor, AP-4, that was identified as a ligand for immunoglobulin kappa promoter E-box elements,(1)suggesting that AP-4 may be related to immunodeficiency diseases. In addition, chromosome 16p13.3 includes a number of genes including the PKD1 gene,(2,3)the autosomal dominant polycystic kidney disease (ADPKD) gene. ADPKD is characterized by progressive development and enlargement of renal cysts.(4)The size and genomic complexity of the PKD1 gene makes it impractical to detect mutations for prenatal diagnosis. Therefore, pedigree-based linkage analysis remains useful for diagnosis of ADPKD. To increase the number of polymorphic markers in the region around AP-4 gene, we performed database searches of 1400 kb of genomic sequence (from contig NT000677 to NT001573: http://www.ncbi.gov/genome/seq.cgi) across the 16p13.3. A number of dinucleotide or tetranucleotide repeats were found, and 20 microsatellites that contain more than 15 contiguous repeats were chosen for further investigation. PMID- 11735305 TI - Lifestyle, minerals and health. AB - This paper proposes that most poor health and behavioral conditions are related to a deficiency of essential minerals or an excess of toxic ones. Clients requesting a hair mineral analysis were required to complete a questionnaire including lifestyle, health status and diet. Most prevalent health conditions in order of frequency were: depression, allergies, low back pain, arthritis, cardiovascular disease and poor digestion. Most deficient minerals were chromium, magnesium, zinc and calcium. Aluminum was the most frequent toxic mineral. it is proposed that such mineral abnormalities were caused by an over consumption of nutrient-poor, highly processed foods such as white flour, sugar and harmful fats. Such a diet promotes poor lifestyles such as smoking, drug abuse, alcoholism and criminal activity. Poor diets and dangerous lifestyles precedes the development of most prevalent diseases. The most common one is cardiovascular disease which increased at an alarming rate in the last century. PMID- 11735306 TI - Vaccines and the risk of insulin-dependent diabetes (IDDM): potential mechanism of action. AB - Immunization with a number of different vaccines, including live and killed vaccines, has been linked to the development of insulin-dependent (type 1) diabetes in humans and animals. Multiple different mechanisms have been proposed to explain the association between vaccines and diabetes. The current paper reviews multiple different mechanisms by which vaccines are known to manipulate the immune system and can induce an autoimmune disease such as type 1 diabetes. Genetic variability may determine which of these pathways, or possible other pathways, predominate in an individual following immunization. PMID- 11735307 TI - The possible crucial role of iron accumulation combined with low tryptophan, zinc and manganese in carcinogenesis. AB - Iron can react with citric acid, interfering with the Krebs cycle, hence with oxidative phosphorylation. Free iron (Fe) can cause considerable oxidative damage both through Fenton reactions and by activating xanthine oxidase, which produces both superoxide (O(2-)) and uric acid (abundant in many cancers). It can also react with lactic acid, reducing its elimination and increasing the acidity of the cytoplasm. Fe can also wreak havoc by reacting with tryptophan, the least abundant and most delicate essential amino acid, which is necessary for the production of serotonin and other substances required by the immune system to fight cancer. On the other hand, in the presence of iron, the tryptophan metabolite quinolinate causes intense lipid peroxidation. Similarly, several other carcinogenic metabolites of tryptophan are particularly dangerous in the presence of Fe. Excess Fe may also interfere with manganese superoxide dismutase and impair the initiation of apoptosis by the mitochondrion, rendering the cells impervious to all the signals to undergo apoptosis from without and from within the cell. Moreover, Fe may also play a crucial role on telomere repair, by activating telomerase. Therefore, by inhibiting apoptosis and enhancing chromosome repair, Fe may bestow immortality upon the cancer cell. Furthermore, Fe is one of the triggers for mitosis. Therefore, increased Fe levels may be essential for the rapid growth characteristic of many malignancies. In turn, the rapid growth further depletes resources from the healthy tissues, exacerbating the deficiencies of the other elements and reducing the ability to fight the malignancy. PMID- 11735308 TI - Corticotropin releasing hormone increases apparent potency of adrenocorticotropic hormone stimulation of cortisol secretion. AB - HYPOTHESIS: Corticotropin releasing hormone (CRH) has a regulatory effect on cortisol secretion in addition to its classic effect of stimulating adrenocorticotropic hormone (ACTH) secretion. REVIEW: There is growing evidence of "long-loop" and paracrine adrenal stimulation by CRH. Data from a study of the ovine-corticotropin releasing hormone (oCRH) stimulation test in 13 sexually abused girls and 13 normal controls was used in Montecarlo simulations of the hypothalamic-pituitary-adrenal axis, to get estimates of adrenal sensitivity to ACTH and cortisol elimination kinetics before and after oCRH administration. In both controls and sexually abused girls, ACTH had an apparent greater effect on cortisol secretion after administration of oCRH compared to its effect during the baseline period. This lends support to the hypothesis and suggests that it should be tested experimentally. PMID- 11735309 TI - The basic reality of mind and spongiform diseases. AB - A change in handedness (chirality) in some amino acids appears to be the basic physical change in degradation-resistant proteins (prions) found in conditions such as Creutzfeldt-Jacob disease (CJD), Alzheimer's disease (AD), bovine spongiform encephalopathy (BSE) and ovine scrapie. The affected structures are primarily innervated by cholinergic nerves. Much evidence suggests that these so called prions (here named chirons) are harmless, non-infectious products. The importance of the cholinergic system allows a new simplified interpretation of these conditions. The main steps are the acetylcholine-cholinesterase splitting of body water with release of free protons in solution, followed by electron dissipation, dioxygen activation and Ca-fluxes. Abiotic physics conserves parity and symmetry by equal amounts of L- and D-forms of molecules. In contrast, the asymmetric pattern of life must be homochiral. Such biomolecules dissolve in water and are thus able to interact in cholinergic hydrodynamics. It is supposed that the instability of the composite weak force by beta-decay causes changes in chirality. These extremely rare events are not frequent enough to explain disease pathology. Experimental, accidental, surgical and abusive inoculations will propagate chirons according to the physical law of self-replication, which also occurs in test tubes without added biological products. Chirons will not be degraded into amino acids in the alimentary canal and will, because they are indigestible, leave the body with the faeces. Chirons are inert also to the immune system and will be engulfed without reaction by phagocytosing cells. They are then stored away in tissues, where they do no harm (if not detected and suspected to be deleterious, thereby causing pathogenic anxiety). The cholinergic system reacts to all kinds of integrity threats and it is this reaction which I propose causes the so-called prion diseases. This pathology seems generally valid, and is here exemplified in AD, CJD, and Kuru disease. It is the cholinergic reaction and not the agent per se that is pathogenic. This is also true of viral infections where the interaction between viral infection and response may explain the enigmatic epidemiology of many neurodegenerative diseases. Intensity and duration of challenges will determine pathophysiology. The new variant of CJD, vCJD, is assumed to result from mutation of a slow virus agent into a more intense variant, which will give disease in younger patients. The pathology is primary protonic, with overactivity in most sub-systems of either enhancing or inhibiting character, but also functional failure or cell death by membrane damage and acidification, for instance in the CNS. The practical results of this proposal will be alleviation of the current BSE crisis. The important main aspects are: chirons are not infectious proteins but inert physical by-products; they are indigestible and not immunogenic, so beef is safe; properly processed and handled meat and bone-meal are not likely to transmit neurodegenerative diseases; chirons cannot even serve as markers in neurologic diseases. PMID- 11735310 TI - BSE did not cause variant CJD: an alternative cause related to post-industrial environmental contamination. AB - The new prion diseases that have emerged in the last 15 years are bovine spongiform encephalopathy (BSE) and variant Creutzfeldt-Jakob disease (variant CJD). Although initially confined to the UK, these diseases have recently emerged in other European countries. The accepted cause of the human disease is that BSE spread from cattle to humans by the consumption of infected beef. However, the evidence that supports this is very thin. This article describes this evidence and lists a series of hypotheses concerning the cause of both BSE and variant CJD. The final hypothesis is based on recent evidence linking prion diseases to environmental factors including manganese. High environmental availability of manganese is associated with the prevalence of those prion diseases not linked to BSE. Therefore it is quite possible that BSE and variant CJD have emerged as a result of manganese-rich industrial pollution that has only occurred in the last century. PMID- 11735311 TI - Neuro-immune-endocrine functional system and vascular pathology. AB - A new interpretation of the response to injury by the nervous, immune and endocrine system is proposed, in order to integrate biochemical knowledge into the respective clinical areas. The discovery that the signaling molecules of the classical nervous, immune and endocrine systems, that is, the neurotransmitters, cytokines and hormones, respectively, are expressed and perceived by the three systems, has enabled us to establish a functional concept of these systems. The hypothetical integration of different pathological processes in a functional response made up by three phases, the immediate or nervous, intermediate or immune and late or endocrine ones, makes it possible to consider that all of them represent different forms of expression of a functional response whose meaning is always the same, that is, inflammation. If the functions that characterize each one of these three phases represent the activity of the nervous, immune and endocrine systems, the biochemical knowledge could be integrated into the functional meaning of each system. PMID- 11735312 TI - Does gastrointestinal Candida albicans prevent ubiquinone absorption? AB - Ubiquinones (coenzyme Qs (CoQ)) are essential for oxidative phosphorylation in yeasts and humans, although the isomers present in each are different. The human coenzyme Q, CoQ10, is administered orally for the treatment of heart disease and other disorders. Some patients, however, require much higher doses than others to attain a therapeutic CoQ10 blood level. We propose that one possible explanation for this variability is Candida colonization of the GI tract. Many common medical treatments including antibiotics and anti-hyperchlorhydric agents increase the risk of GI tract Candida colonization. Subsequent uptake and utilization of supplemental CoQ10 by the yeast could diminish availability for the human subject. Data from one patient and an in vitro pilot study using two pathogenic strains of C. albicans support this hypothesis. If C. albicans in the GI tract can hinder availability and interfere with therapeutic effects of CoQ10, it could be of clinical significance for large numbers of patients. PMID- 11735313 TI - Bovine spastic paresis: etiological hypotheses. AB - Bovine spastic paresis (BSP) is a rare and little-known disease of cattle. The etiology remains unknown. A pathogenesis close to the sub-acute transmissible spongiform encephalopathies (TSEs) has repeatedly been advanced. In order to confirm an infectious origin, intra- and inter-species transmission studies are proposed. A positive result would bring a major advance in knowledge. PMID- 11735314 TI - Sudden infant death syndrome and enteric infection. AB - The association of Helicobacter pylori in the stomach, trachea and lungs with the incidence of SIDS, gastric ulcers and cancer may have a counterpart in animals. In field studies of white muscle disease (WMD) and hepatic necrosis in selenium deficient pigs dying suddenly, veterinarians identified gastric ulcers in 40% of inspected piglets. The lesion was also commonly observed by researchers in experimentally produced vitamin E-selenium deficiency and other researchers suspected that gastric ulcers in swine may be associated with vitamin E-selenium deficiency. Mice preferentially concentrated (75)selenium in peritoneal exudative cells (PEC) when (75)selenium as selenium selenate was administered by stomach tube to selenium-deficient mice. Selenium concentrated in PECs as glutathione peroxidase (GSHP(x)). GSHP(x)-deficient leucocytes in peritoneal exudate failed to kill yeast cells. GSHP(x) deficiency has also been associated with decreased microbicidal activity of leucocytes in patients with chronic granulomatosis. The selenium-deficient swine were usually growing rapidly in crowded conditions, and, apart from WMD and hepatic necrosis, edema was prominent in the spiral colon, subcutaneous tissues, lungs and submucosa of the stomach. The elevated immunological response in the spleen and lungs of SIDS victims suggests an initial defective microbicidal propensity of the peritoneal exudative cells. PMID- 11735315 TI - An immune hypothesis of sexual orientation. AB - Sexual orientation is encoded within immune-cell subsets (ICS) of mucosal and epithelial tissues. Gender orientation may be encoded within other ICS. Many immune cells: recognize and react to H-Y and H-X antigens; and enact these perceptions and reactions in accord with the perceiver's and the perceived's MHC haplotype, XX or XY status, and immune-self recognition. Non-heterosexual orientations derive from excessive cross-priming, accompanied by clonal deletions, clonal expansions, anergy and tolerance. For at least some tissues, cross-priming sufficient to induce altered orientations may occur during critical periods of immunological development and can occur during fetal and infant development via maternal-fetal transfusion, placental pathology, and impaired maternal nutrient-status or via excessive peripheral apoptosis during postnatal illness. Mast cell interactions with neurons illustrate how mucosal perceptions can be transduced into neuronal signals that modulate CNS events. This hypothesis is testable by mixed-lymphocyte reactions in appropriate cell subsets. Dendritic cell immunizations are a potential therapy. PMID- 11735316 TI - Carbon monoxide: the elusive environmental toxicant. AB - The pivotal role of carbon monoxide (CO) in understanding the pathology of diseases is long overdue. The significance of this odourless, inert gas was trivialised throughout the years in advancement of corporate interest, with over riding emphasis on economic gains. Thus, under the auspices of advancement of socioeconomic interest, the role of carbon monoxide, an extremely dangerous biproduct of incomplete combustion of any hydrocarbon, was inevitably obscured by the lure of profitability. Although odourless and inert, CO is a potent chemical asphyxiant whose primary toxic action is through hypoxia. It is on the basis of its prolonged effects at cellular level, by alteration to redox state of each and every single living cell, that, the author argues, it contributes to the ever increasing levels of neurodegenerative, autoimmune, maladaptive, behavioural syndromes known to be especially prevalent within industrialised western societies. PMID- 11735317 TI - Cholesterol: an important but relatively overemphasized risk factor for ischemic heart disease. AB - Educational messages directed at the public to prevent ischemic heart disease (IHD) are generally based on cholesterol-reduction. However, IHD has multiple risk factors, and a study was performed to help determine whether or not the allocation of educational messages among risk factors is appropriate: The severity of high cholesterol was compared with the severity of multiple other major risk factors for IHD, and the beneficial effects of cholesterol-reduction was compared with the benefits of multiple other major preventative factors for IHD. It was found that high cholesterol levels, and multiple other risk factors, generally give a risk of around 2.0 for developing IHD. Cholesterol-reduction by statins, and multiple other factors which prevent IHD, generally reduce the risk of IHD by about 30-40%. It was concluded that the allocation of educational messages to reduce the incidence of IHD should significantly increase discussions of non-cholesterol risk and preventative factors. PMID- 11735318 TI - Possible role of brain thyroid hormones in the effects of bright light on mood and behavior. AB - Considerable evidence suggests that light may significantly affect the metabolism of cerebral thyroid hormones. Changes in the brain thyroid economy, independent of peripheral changes in thyroid status, may produce considerable behavioral effects. Exposure to bright light affects the mood and behavior of healthy people and individuals with psychiatric disorders. The author suggests that the effects of bright light on mood and behavior may be partly mediated by light-induced changes in the metabolism of brain thyroid hormones. PMID- 11735319 TI - Possible genetic link between eating disorders and seasonal changes in mood and behavior. AB - Studies suggest that there is a seasonal pattern of mood fluctuations and eating behavior in patients with eating disorders. Multiple lines of evidence suggest that serotonergic pathways are involved in the mechanisms of eating disorders and seasonal changes in mood and behavior. Researchers have investigated whether variants of genes related to serotonergic transmission are associated with seasonal affective disorder (SAD) and eating disorders. There is evidence that the -1438G/A promoter polymorphism of the 5-HT2A gene plays a role in the development of SAD and eating disorders. Variation of the tryptophan hydroxylase gene may play a part in eating behavior and weight regulation in females with SAD. The author suggests that there may be a genetic link between SAD and eating disorders. It is possible that there are specific inherited personality types with a predisposition to both eating disorders and SAD. PMID- 11735320 TI - Role of endogenous opioids in the effects of light on mood and behavior. AB - Skin cells produce endogenous opioids in response to light. This results in increased skin and plasma levels of endogenous opioids. Increased plasma levels of endogenous opioids may improve mood and affect behavior. The author suggests that improvement in mood after exposure to light may be related to the light induced increase in levels of endogenous opioids. The author speculates that genetic factors may affect the response of skin cells to light: different people may have different genetically determined responses to natural or artificial light. The author also suggests that light may affect endogenous opioids via several different mechanisms. PMID- 11735321 TI - Oral vaccination with autologous platelets in chronic autoimmune thrombocytopenic purpura. AB - Idiopathic thrombocytopenic purpura (ITP) is a common disease of autoimmune mechanism in which nearly 40% of patients need a third-line treatment after steroids and splenectomy failure. Due to the variable results of the multiple therapeutic modalities in refractory patients it is necessary to develop a more effective treatment. Following previous assays of immunotolerance with the administration of "oral vaccines", we propose a therapeutic protocol in chronic ITP patients with autologous cryopreserved or liophilized platelets administered in enteric capsules. The procedure is theoretically feasible and technically easy and does not need the isolation of the specific antigen target of the autoantibodies in each particular case. PMID- 11735322 TI - Age-associated neovasculopathy with recurrent bleeding. AB - We hypothesize the existence of an age-associated neovasculopathy with recurrent bleeding. It could be the result of age-related defects in the regulation of the angiogenetic process creating pathologic small vessels with a bleeding tendency. Conditions like subdural hematoma, ocular angiopathies, intestinal angiodysplasia, coronary atherosclerotic plaques and others, may be the clinical expression of such a neovasculopathy. PMID- 11735323 TI - Mechanism of anesthetic action: oxygen pathway perturbation hypothesis. AB - Although more than 150 years have passed since the discovery of general anesthetics, precisely how they work remains a mystery. We propose a novel unitary mechanism of general anesthesia verifiable by experiments. In the proposed mechanism, general anesthetics perturb oxygen pathways in both membranes and oxygen-utilizing proteins, such that the availability of oxygen to its sites of utilization is reduced, which in turn triggers cascading cellular responses through oxygen-sensing mechanisms, resulting in general anesthesia. Despite the general assumption that cell membranes are readily permeable to oxygen, existing publications indicate that these membranes are plausible oxygen-transport barriers. The present hypothesis provides a unified framework for explaining phenomena associated with general anesthesia and experimental results on the actions of general anesthetics. If verified by experiments, the proposed mechanism also has other significant medical and biological implications. PMID- 11735324 TI - Catecholamine metabolism in the brain by membrane-bound and soluble catechol-o methyltransferase (COMT) estimated by enzyme kinetic values. AB - A kinetic model was constructed to reevaluate the catecholamine metabolism in hypothetical brain homogenates. Earlier published kinetic values of recombinant membrane-bound (MB-) COMT and soluble (S-) COMT were combined with data suggesting that MB-COMT represents 70% and 30% of total COMT protein in human and rat brain, respectively. In the rat brain model L-DOPA and 3,4-dihydroxybenzoic acid were O-methylated mainly via S-COMT, while dopamine and noradrenaline, at low concentrations, were O-methylated slightly more by MB-COMT. In the human brain model dopamine and noradrenaline were metabolized primarily by MB-COMT. The ratio of meta (3-methoxy) over para (4-methoxy) product formation from 3,4 dihydroxybenzoic acid was higher for MB-COMT than S-COMT. It is suggested that MB COMT clearly predominates the O-methylation of dopamine and noradrenaline also in vivo. Additionally, meta/para ratios could support the enrichment of either isoform of COMT in a homogenate sample. PMID- 11735325 TI - Medicine may be reducing the human capacity to survive. AB - It appears that limited natural selection is taking place in populations of developed countries, since most individuals survive and have the full opportunity to reproduce. This paper addresses contemporary natural selection in a developed country (Australia) using the biological state index. Although the general context of this paper focuses on Australia it can be expected that most other first-world and/or developed countries follow a similar pattern. The findings of this study, that 98% of individuals survive through their reproductive period and have the full opportunity to reproduce, support predictions that natural selection has limited influence on the evolution of first-world populations. It appears that first-world populations may not be naturally well adapted to their environment but use medical treatments/technology to increase their survival capacity and maintain fitness. This has two apparent consequences. First, the fitness of individuals will decrease, since less favorable genes can accumulate in the population, and secondly, disease processes will remain fit as they adapt to the selective pressures exerted by medicine. If medical treatment becomes ineffective, extensive mortality is expected since fit disease processes will be unleashed on unfit human populations. It appears that a possible answer to these problems may be found in gene therapy. PMID- 11735326 TI - Onset of spontaneous pneumothorax and the synodic lunar cycle. AB - The relation between spontaneous pneumothorax events and synodic lunar cycle was studied in a retrospective analysis of patients with a first or recurrent spontaneous pneumothorax. The study included a total of 244 patients, 203 males and 41 females. The data were arranged along the first (new moon) to 28th day (one day before another new moon) of the synodic lunar cycle. Periodogram analysis revealed a 14-day rhythm, significant for the male and pooled sample. Cosinor analysis found the whole synodic lunar cycle and its 2nd, 4th and 5th harmonics as significant. Maximal accumulation of cases happened 1 week before and 1 week after the new moon. Mechanisms of a putative moon influence are not clear. PMID- 11735327 TI - Retrograde embolism and atherosclerosis development in the human thoracic aorta: are the fluid dynamics explanations valid? AB - The mechanisms of atherosclerosis development in the thoracic aorta is miscellaneous and still only partially understood. The marked variability of the sites of deposition of atherosclerotic plaques in the aorta could not be clarified based solely on the risk-factors theory of atherosclerosis. The sites of deposition of atherosclerotic plaques are considered to be affected by blood flow patterns that cause areas of altered shear stress on the aortic wall. Close relations between protruding aortic plaques (PAP), stroke and peripheral emboli were established. The analysis of PAP distribution and motion to characterize atherogenesis in the human thoracic aorta and the pathogeneses of embolic events was performed. We concluded that protruding aortic plaques and markers of relative aortic flow instability (occurrences of vortices) are predominantly noticed in the human arch and in the descending aorta, whereas the ascending aorta showed lesser prevalence of atheromatosis. Reversal and rotational blood flow in the thoracic aorta most likely exist in all patients with systemic emboli and mobile protruding aortic atheromas. Therefore, retrograde cerebral embolism from distal aortic plaques is conceivable. PMID- 11735328 TI - Stability-variability conflict as a reason for cancer: physical and social aspects. AB - We present an analysis of cancer based on the synthesis of physical and social models of life as the stability-variability conflict (SVC). We showed that the intensity of the SVC increases with an increase in the rate of evolution or progress of species and populations. The probability of oncogenesis depends on this intensity and on the gray zone in the host's homeostasis. An organism and its cells are two interdependent systems which in extreme situations become antagonistic and competitive. The hierarchy of the biosystems (cell, organism, population) does not resolve the SVC and does not decrease the gray zone of this conflict. Cancer develops mainly due to external effects and results in some internal, including genomic, alterations which result in cell death (apoptosis), or cause cells to begin a new stage in their lives, possibly leading to tumors. In this respect, cancer is a rescuer of the cells, although it brings about the death of the host. An increase in the production of tumor-associated proteins in cancer patients is an example of such changes. Cancer can also be analysed in terms of entropy and information. In the context of the SVC, cancer is irreversible deviations from the "golden mean" solution. The golden mean and cancer deviation are considered universal categories of physical and social reflections of the SVC as the quintessence of life. PMID- 11735329 TI - Transdifferentiation of neoplastic cells. AB - Transdifferentiation is a process in which a stable cell's phenotype changes to that of a distinctly different cell type. It occurs during certain physiological processes and leads to transition of tumor cell phenotypes. The latter process includes neoplastic epithelial-epithelial transition, neoplastic epithelial mesenchymal transition, neoplastic mesenchymal-epithelial transition and transition between non-neural and neural neoplastic cell. This phonomenon is exemplified in some origin-debated tumors, such as carcinosarcoma, pleomorphic adenoma, synovial sarcoma, Ewing's/pPNET, and malignant fibrohistiocytoma. We propose that differentiation disturbance of cancer cells should include not only undifferentiation and dedifferentiation, but also transdifferentiation as well. Tumor cell transdifferentiation may be influenced or determined by cellular genetic instabilities, proliferation and apoptosis, as well as by extracellular matrix and growth factors. PMID- 11735332 TI - Lentivirus in the clinic. PMID- 11735334 TI - You say tomato and I say tomahto: human genetics and gene therapy. PMID- 11735335 TI - Gene therapy for acute diseases. AB - The use of gene transfer systems to study cell function makes it apparent that overexpression of a transgene can restore or improve the function of a protein and positively influence cell function in a predetermined manner for purposes of counterbalancing cellular pathophysiology. The ability of some gene transfer vehicles to produce transgene product within hours of delivery positions gene transfer as a unique pharmaceutical administration system that can quickly affect production of biologic response modifiers in a highly compartmentalized fashion. This approach can be expected to overcome many of the adverse effects and high costs of systemic delivery of recombinant pharmaceuticals. This review highlights recent advances toward development of gene therapies for acute illnesses with particular emphasis on preclinical models of disease. In this context, a growing body of data suggests that gene therapies for polygenic and non-genetic diseases such as asthma, cardiogenic and non-cardiogenic pulmonary edema, stroke, subarachnoid hemorrhage, seizures, acute myocardial infarction, endovascular thrombosis, and infections may someday be options for the treatment of patients. PMID- 11735336 TI - Immune evasion by muscle-specific gene expression in dystrophic muscle. AB - Muscle tissue from Duchenne muscular dystrophy patients and the Dmd(mdx/mdx) (hereafter referred to as mdx) mouse is characterized by an abundance of necrotic myofibers and infiltrating macrophages. Both features may provide additional stimulus to the immune response directed against novel antigens, such as those delivered by gene therapy vectors. It has previously been shown that the immune evasion achieved by adeno-associated virus in healthy muscle fails in one model of muscular dystrophy. Here, we examined the immune response to adenoviral vectors and their transgenes in normal and mdx mice. We found that mdx mouse muscles contain 20 times more macrophages and 7 times more dendritic cells than healthy muscles. This higher professional antigen-presenting cell content results in a stronger immune response to antigens that can be directly presented by those cells, including viral antigens and constitutively expressed transgene products. However, we did not detect a significant immune response to beta-galactosidase expressed specifically in muscle, even at high expression levels. This result suggests that cross-presentation is not more effective in mdx mouse muscle, and that targeted vectors and tissue-specific promoters may be useful tools for evasion of the immune response in dystrophic muscle. PMID- 11735337 TI - Ablating adenovirus type 5 fiber-CAR binding and HI loop insertion of the SIGYPLP peptide generate an endothelial cell-selective adenovirus. AB - Adenovirus type 5 (Ad) based vectors transduce vascular endothelial cells (EC) and have been widely used for vascular gene transfer. However, many cell types express the Ad receptor (cox-sackievirus adenovirus receptor; CAR), preventing selective EC infection and precluding clinical use. We previously isolated the human EC-binding peptides SIGYPLP and LSNFHSS by phage display and demonstrated by means of a bispecific antibody that SIGYPLP directs efficient, high-level, EC selective Ad-mediated gene transfer. We now generate genetically modified Ad fiber proteins with selective EC tropism by engineering these peptides into the HI loop of the Ad fiber. SIGYPLP, but not LSNFHSS, enhanced EC selectivity, demonstrating maintenance of peptide-cell binding fidelity upon incorporation into virions. Combining fiber mutations that block CAR binding (detargeting) with SIGYPLP insertion (retargeting) generated a novel Ad vector, AdKO1SIG, in a single component system. AdKO1SIG demonstrated efficient and selective tropism for EC compared with control Ad vectors. This is the first demonstration of genetic incorporation of a novel, mammalian, cell-selective ligand that retains its targeting fidelity in the Ad fiber HI loop, in combination with point mutations that abolish fiber-CAR interaction. This study demonstrates the potential for improving the cell-selectivity and safety of adenoviral vectors. PMID- 11735338 TI - Viral IL-10 gene transfer inhibits DTH responses to soluble antigens: evidence for involvement of genetically modified dendritic cells and macrophages. AB - Expression of the viral interleukin-10 (vIL-10) gene within one joint of an animal with polyarticular, inflammatory arthritis suppresses disease in both treated and untreated joints (the "contralateral effect"). We used a mouse delayed-type hypersensitivity (DTH) model to investigate this phenomenon. Adenoviral delivery of the vIL-10 gene suppressed DTH reactions in injected and contralateral paws. T lymphocytes recovered from immunized mice injected with the adenoviral vector (ad-vIL-10) were unable to transfer the DTH response, but were not inhibitory. Peritoneal exudate cells recovered from mice injected intraperitoneally with ad-vIL-10 inhibited DTH reactions in recipient mice, but only when the donor mice had been sensitized to the antigen used to incite the DTH response. Dendritic cells (DCs) recovered from the draining lymph nodes of mice injected with ad-vIL-10 behaved similarly. Bone-marrow-derived DCs cultured ex vivo with ad-vIL-10 or recombinant mouse IL-10 also suppressed DTH reactions by adoptive transfer when pulsed with the inciting antigen. Collectively, these data suggest a mechanism for the contralateral effect in which genetically modified macrophages and DCs present antigen in the context of high, local concentrations of vIL-10, thereby generating unresponsive T lymphocytes. These findings suggest new ways in which to treat immune-driven diseases by gene and cell therapy. PMID- 11735339 TI - Gene therapy for head and neck cancer using vaccinia virus expressing IL-2 in a murine model, with evidence of immune suppression. AB - We evaluated the efficiency of recombinant vaccinia virus expressing interleukin 2 (rvv-IL-2) as a tumor vaccine in an immunocompetent mouse model of head and neck squamous cell carcinoma (SCC VII/SF). Mice with five-day-old tumors in the floor of the mouth were treated with rvv-IL-2 by intratumoral injections. These treated mice survived longer (P <.03) than mice treated with control vaccines. Splenocytes, bone marrow, and lymph node cells from tumor-bearing mice responded poorly to concanavalin A stimulation, suggesting induction of immunosuppression. The rvv-IL-2 virus grew for 7 days in the tumor following intratumoral injection. We did not detect any virus particles in several normal organs following rvv-IL-2 injection. Comparison of expression levels of several potential immune inhibitory mediators between the tumors growing in mice and cultured tumor cells demonstrated higher expression of IL-10, GM-CSF, TGF-beta, and NO synthetase in tumors. These results suggested possible roles for these molecules in immunosuppression. We conclude that rvv-IL-2 has potential as a therapeutic vaccine for head and neck cancer and that it can be more effective provided the immunosuppression is reversed. PMID- 11735340 TI - Immediate and long-term safety of recombinant adeno-associated virus injection into the nonhuman primate muscle. AB - Previous studies on distribution and toxicity of viral vectors administered in monkeys indicated that the nonhuman primate model has a reasonable predictive value for clinical applications. In this study, eight macaques were injected intramuscularly with recombinant adeno-associated virus (rAAV) at doses similar to those administered to hemophilia B patients, and followed to analyze the dissemination and shedding in biological samples and long-term persistence in distant organs. Following rAAV delivery, we found vector genome in various biological fluids for up to 6 days and infectious particles exclusively in the serum during the first 48-72 hours. rAAV sequences were detected in peripheral blood mononuclear cells (PBMC) for up to 10 months. At necropsy, 8 to 18 months after rAAV delivery, rAAV sequences were found in lymph nodes and livers but never in the gonads. Tissue examination, of liver in particular, showed no abnormalities. We concluded that during our experimental time frame, rAAV mediated gene transfer into skeletal muscle of macaques seemed to be safe with respect to the recipient and the environment. However, it was associated with a transient viremia and the persistence of rAAV sequences in PBMC, lymph nodes, and liver, the long-term consequences of which remain unknown. PMID- 11735341 TI - Generation of species cross-reactive aptamers using "toggle" SELEX. AB - Species cross-reactivity facilitates the preclinical evaluation of potentially therapeutic molecules in animal models. Here we describe an in vitro selection strategy in which RNA ligands (aptamers) that bind both human and porcine thrombin were selected by "toggling" the protein target between human and porcine thrombin during alternating rounds of selection. The "toggle" selection process yielded a family of aptamers, all of which bound both human and porcine thrombin with high affinity. Toggle-25, a characteristic member, inhibited two of thrombin's most important functions: plasma clot formation and platelet activation. If appropriate targets are available, the toggle strategy is a simple measure that promotes cross-reactivity and may be generalizable to related proteins of the same species as well as to other combinatorial library screening strategies. This strategy should facilitate the isolation of ligands with needed properties for gene therapy and other therapeutic and diagnostic applications. PMID- 11735342 TI - The functional and structural outcome of inner ear gene transfer via the vestibular and cochlear fluids in mice. AB - Mice present an ideal model for inner ear gene therapy because their genome is being rapidly sequenced, their generation time is relatively short, and they serve as a valuable model for human hereditary inner ear disease. However, the small size of the mouse inner ear poses a particular challenge for surgical procedures. We have developed a new approach for viral inoculation into the mature mouse inner ear, using a replication-deficient adenovirus expressing the bacterial gene lacZ. We administered the virus through the posterior semicircular canal (canalostomy) and into the cochlea (cochleostomy). Both approaches caused lacZ to be expressed in cells lining the perilymphatic space. One canalostomy case showed gene expression in sensory cells of the crista ampullaris, whereas the cochleostomy group showed gene expression in the sensory cells in the organ of Corti and saccule. Functional tests after the surgery showed that the canalostomy preserved hearing, whereas the cochleostomy did not. Any vestibular function transiently lost after the canalostomy was recovered. Our findings indicate that inoculation of adenovirus vectors into the mouse inner ear through the semicircular canal has the potential to efficiently introduce transgenes to the vestibular system and the cochlea without compromising hearing. PMID- 11735343 TI - Lack of germline transmission of vector sequences following systemic administration of recombinant AAV-2 vector in males. AB - A potential consequence of systemic administration of viral vectors is the inadvertent introduction of foreign DNA into recipient germ cells. To evaluate the safety of in vivo recombinant adeno-associated virus (rAAV) mediated gene transfer approaches for hemophilia B, we explored the risk of germline transmission of vector sequences following intramuscular (IM) injection of rAAV in four species of male animals (mouse, rat, rabbit and dog). In vector biodistribution studies in mice and rats, there is a dose-dependent increase in the likelihood that vector sequences can be detected in gonadal DNA using a sensitive PCR technique. However, in dogs DNA extracted from semen is negative for vector sequences. To address this discrepancy, studies were done in rabbits, and both semen and testicular DNAs were analyzed for the presence of vector sequences. These studies showed that no AAV vector sequences were detected in DNA extracted from rabbit semen samples collected at time points ranging from 7 to 90 days following IM injection of 1 x 10(13) vector genomes rAAV (vg) per kg. In contrast, DNA extracted from gonadal tissue was positive for vector sequences, but the positive signals diminished in number and strength with time. By FISH analysis, AAV signals were localized to the testis basement membrane and the interstitial space; no intracellular signal was observed. We observed similar findings following hepatic artery administration of rAAV in rats and dogs, suggesting that our findings are independent of the route of administration of vector. Attempts to transduce isolated murine spermatogonia directly with AAV lacZ were unsuccessful. In clinical studies human subjects injected IM with an AAV vector at doses up to 2 x 10(12) vg/kg have shown no evidence of vector sequences in semen. Together, these studies suggest that rAAV introduced into skeletal muscle or the hepatic artery does not transduce male germ cells efficiently. We conclude that the risk of inadvertent germline transmission of vector sequences following IM or hepatic artery injection of AAV-2 vectors is extremely low. PMID- 11735344 TI - Regulated, adenovirus-mediated delivery of tyrosine hydroxylase suppresses growth of estrogen-induced pituitary prolactinomas. AB - Prolactin-secreting adenomas are one of the most common types of intracranial neoplasm found in humans. The modalities of clinical treatment currently in use include D(2)-dopamine receptor agonists, surgery, and radiotherapy, and the success rates for treatment are good. However, there are prolactinomas that are difficult to treat. As an alternative, we have developed a gene therapy strategy in which the rate-limiting enzyme in dopamine synthesis, tyrosine hydroxylase (TH), is overexpressed in the anterior pituitary (AP) gland. Because dopamine is known to have an inhibitory effect on lactotroph growth and prolactin secretion, we developed a system that would enable its local synthesis from freely available precursor amino acids. A dual adenovirus tetracycline-regulatable expression system was generated to control the production of TH. In the absence but not presence of the tetracycline analog doxycycline, TH expression was observed in AP tumor cell lines AtT20, GH3, and MMQ. In both primary AP cell cultures and the AP gland, in situ expression of TH was seen in lactotrophs, somatotrophs, corticotrophs, thyrotrophs, and gonadotrophs in the absence but not presence of doxycycline. The ability of this system to inhibit hyperprolactinemia and pituitary lactotroph hyperplasia was then assessed in a model of estrogen- or estrogen/sulpiride-induced pituitary tumors. In the absence but not presence of doxycycline, a 49% reduction in pituitary growth and 58% reduction in the increase of circulating prolactin levels were observed in estrogen, but not estrogen/sulpiride, treated rats. These results indicate that in situ dopamine enhancement gene therapy can be a useful tool for the treatment of prolactinoma. Dopamine synthesis can be tightly regulated and the therapeutic benefit of the system is only inhibited when local dopamine signaling is impaired. PMID- 11735345 TI - Absence of germline infection in male mice following intraventricular injection of adenovirus. AB - The possibility of inadvertent exposure of gonadal tissue to gene therapy vectors has raised safety concerns about germline infection. We show here that the receptor for coxsackie B viruses and adenoviruses 2 and 5 (CXADR) is expressed in mouse germ cells, suggesting the possibility that these viruses could infect germ cells. To directly assess the risk of germline infection in vivo, we injected an adenovirus carrying the germ-cell-specific protamine promoter fused to the bacterial lacZ reporter gene into the left ventricular cavity of mice and then monitored expression of the reporter gene in germ cells. To differentiate between infection of stem cells and differentiating spermatogenic cells, we analyzed expression of the reporter cassette at different times after viral delivery. Under all conditions tested, mice did not express the Escherichia coli beta galactosidase protein in developing spermatids or in mature epididymal spermatozoa. Primary germ cells cultured in vitro were also refractory to adenoviral infection. Our data suggest that the chance of vertical germline transmission and insertional mutagenesis is highly unlikely following intracoronary adenoviral delivery. PMID- 11735346 TI - Lentiviral vector gene transfer into fetal rhesus monkeys (Macaca mulatta): lung targeting approaches. AB - We previously reported the efficiency of gene transfer in fetal monkeys using retroviral vectors and an intraperitoneal (IP) approach. Here, we explored intrapulmonary administration to determine whether gene transfer can be limited to the developing lung. The HIV-1-derived lentiviral vector (VSV-G pseudotyped; 1 x 10(7) infectious particles/fetus), using the enhanced green fluorescent protein (EGFP) as a reporter, was directly injected into fetal lung with ultrasound guidance (n=4; 55 or 70 days gestation; term 165+/-10 days). Fetuses were monitored sonographically, fetal/maternal blood samples collected during gestation, and four of four healthy newborns were delivered at term. All lung lobes were positive for the transgene (< or = 1%) when assessed by PCR, and transgene expression was observed by direct fluorescence microscopy and flow cytometry. The results of this study show the following: (1) successful gene transfer in fetal monkeys using an intrapulmonary approach; (2) less transduction of non-pulmonary tissues with gene transfer at 70 days gestation compared with 55 days gestation or use of an IP approach; (3) that the pulmonary epithelium was EGFP-positive by immunohistochemistry; and (4) no evidence of transplacental transport of vector sequences or antibody responses in the dams. The results of these investigations indicate the efficiency of fetal gene transfer by intrapulmonary delivery, and emphasize the importance of the fetal monkey as a preclinical model system for exploring in utero genetic treatment strategies for pulmonary disorders. PMID- 11735347 TI - Glial cell line derived neurotrophic factor delays photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa. AB - We designed experiments to evaluate the therapeutic potential of glial cell line derived neurotrophic factor (GDNF) to rescue photoreceptors from genetically determined cell death. Gene transfer of the neurotrophic factor to the retina was achieved via a recombinant adeno-associated virus (rAAV) vector containing the chicken beta-actin promoter/immediate early cytomegalovirus enhancer (CBA) driving the human GDNF gene. We delivered AAV-CBA-GDNF to the retinas of an animal model of retinitis pigmentosa, the TgN S334ter-4 rhodopsin line of transgenic rats. Immunohistochemical studies localized AAV-CBA-GDNF-derived recombinant protein to cell bodies, inner segments, and outer segments of photoreceptor cells as well as to retinal pigment epithelial cells. We assessed the effect of viral delivery by morphometric and electroretinographic analysis. These experiments showed that GDNF vector treatment leads to increased rod photoreceptor survival as indicated by morphometric analysis of outer nuclear layer thickness. AAV-CBA-GDNF-treated retinas also demonstrated functional improvement by the substantially increased amplitude of electroretinograms. AAV CBA-GDNF delivery had a significant rescue effect on photoreceptor degeneration in this animal model. PMID- 11735348 TI - Dual acting anti-inflammatory drugs: a reappraisal. AB - Rheumatic diseases are the most prevalent causes of disability in western countries, and non-steroidal anti-inflammatory drugs (NSAIDs) are still the most commonly used remedies. However, NSAIDs cause several serious adverse effects, the most important being from gastric injury to gastric ulceration and renal damage. Attempts to develop non-steroidal anti-inflammatory remedies devoid of these shortcomings-especially gastrointestinal toxicity-have followed several strategies. Non-steroidal anti-inflammatory drugs have, therefore, been associated with gastroprotective agents that counteract the damaging effects of prostaglandin synthesis suppression; however, a combination therapy introduces other problems of pharmacokinetics, toxicity, and patient's compliance. More recently, incorporation of a nitric oxide (NO)-generating moiety into the molecule of several NSAIDs was shown to greatly attenuate their ulcerogenic activity; however, several findings suggest a possible involvement of NO in the pathogenesis of arthritis and subsequent tissue destruction. A most promising approach seemed to be the preparation of novel NSAIDs, targeted at the inducible isoform of prostaglandin synthase (COX-2); they appear to be devoid of gastrointestinal toxicity, in that they spare mucosal prostaglandin synthesis. However, a number of recent studies have raised serious questions about the two central tenets that support this approach, namely that the prostaglandins that mediate inflammation and pain are produced solely via COX-2 and that the prostaglandins that are important in gastrointestinal and renal function are produced solely via COX-1. So, a growing body of evidence shows that COX-2 (not only COX-1) also plays a physiological role in several body functions and that, conversely, COX-1 (not only COX-2) may also be induced at sites of inflammation. More recent and puzzling data shows that COX-2 is induced during the resolution of an inflammatory response, and at this point it produces anti-inflammatory (PGD2 and PGF2alpha), but not proinflammatory (PGE2) prostaglandins; inhibition of COX-2 at this point thus results in persistence of the inflammation. Moreover, COX-2 selective NSAIDs have lost the cardiovascular protective effects of non selective NSAIDs, effects which are mediated through COX-1 inhibition (in addition, COX-2 has a role in sustaining vascular prostacyclin production). The generation of other very important products of the arachidonic acid cascade (besides cyclooxygenase-produced metabolites) is inhibited neither by non selective nor by COX-2 selective NSAIDs. The products generated by the 5 lipoxygenase pathway (leukotrienes) are particularly important in inflammation; indeed, leukotrienes increase microvascular permeability and are potent chemotactic agents. Moreover, inhibition of 5-lipoxygenase indirectly reduces the expression of TNF-alpha (a cytokine that plays a key role in inflammation). These data and considerations explain the efforts to obtain drugs able to inhibit both 5-lipoxygenase and cyclooxygenases, the so-called dual acting anti-inflammatory drugs. Such compounds retain the activity of classical NSAIDs, while avoiding their main drawbacks, in that curtailed production of gastroprotective prostaglandins is associated with a concurrent curtailed production of the gastro damaging and bronchoconstrictive leukotrienes. Moreover, thanks to their mechanism of action, dual acting anti-inflammatory drugs could not merely alleviate symptoms of rheumatic diseases, but might also satisfy, at least in part, the criteria of a more definitive treatment. Indeed, leukotrienes are pro inflammatory, increase microvascular permeability, are potent chemotactic agents and attract eosinophils, neutrophils and monocytes into the synovium. PMID- 11735349 TI - Anti-inflammatory capabilities of macrolides. AB - Macrolide antibiotics play a significant role in clinical practise due not only to their antibacterial activity, but also to their accompanying anti-inflammatory effect that is independent of their antibiotic action. Several studies reported in literature show that macrolides affect several inflammatory processes, such as migration of neutrophils, the oxidative burst in phagocytes and production of pro inflammatory cytokines, although the precise mechanisms are not clear. They also inhibit eosinophilic inflammation and may be useful in the treatment of patients with steroid-dependent asthma. Macrolides are also effective in diffuse panbronchiolitis, chronic sinusitis and inflammatory skin diseases. PMID- 11735350 TI - The effect of aprotinin on ischemia-reperfusion injury in the rabbit kidney. AB - Tissue subjected to a period of ischemia undergoes functional and morphological damage that increases during the reperfusion phase. In this study, the protective effect of aprotinin, which is a protease inhibitor, was assessed in a rabbit unilateral renal ischemia-reperfusion (I/R) model. New Zealand rabbits, weighing 1.5-2 kg, were randomized to receive either aprotinin 30.000 KIU x kg(-1) and 10.000 KIU x kg(-1) x h(-1) i.v. infusion (group I, n= 7) or equivalent volumes of 0.09% sodium chloride (SF) (group II, control, n= 7) i.v. 15 minutes before a 45 minutes interruption of left renal artery blood flow and then 45 minutes of reperfusion. Blood samples were obtained before and after the ischemia reperfusion period for measurement of nitric oxide serum (NO) levels with the nitrite/nitrate colorimetric method. Histological changes were evaluated by quantitative measurements using a numerical score (0-4) and immunohistochemical analysis of inducible nitric oxide synthase (iNOS) expression was determined. A Wilcoxon W -test was used for statistical analysis of biochemical measurements and mean values were expressed as +/-sd. Histological examination revealed the distinctive pattern of ischemic renal tissue injury with obvious signs of epithelial necrosis. The intensity of epithelial necrosis was more extensive in the SF group. Immunohistochemical analysis showed that there was severe immunostaining in the tubular epithelium in both cortical and medullary regions and iNOS expression was more intense in SF-only cases. The staining results for aprotinin cases did not differ much from the non-ischemic kidney. Biochemical analysis revealed an increase in serum NO levels in both groups (P< 0.05), but this was more evident in the SF group (mean NO levels were 38.63 +/- 19.03 micromol x L(-1) in group I, 50.63 +/- 24.28 micromol x L(-1) in group II). No statistically important difference was observed between the two groups. These results suggest that aprotinin may be beneficial in the prevention of systemic inflammation after transient renal ischemia. PMID- 11735351 TI - Second generation antihistamines in the treatment of seasonal allergic rhinitis due to Parietaria and cypress pollen. AB - Second generation antihistamines have been employed in the treatment of seasonal allergic rhinitis for many years. However, their effects on two distinctive Mediterranean allergic conditions, viz. Parietaria pollinosis and cypress pollinosis, have been scarcely investigated, so far. A comparative efficacy and side effect trial of astemizole and terfenadine in the treatment of seasonal allergic rhinitis due to either Parietaria or cypress pollen was carried out in 27 adult patients, according to a double-blind, double-dummy parallel-group design. Airborne pollen monitoring allowed comparison of symptom scores with pollen counts. Seven patients (26%) withdrew, due to poor symptom control. In contrast, in a subset of 15 patients who completed the trial, treatment led to a substantial and statistically significant decline in symptom severity in both the astemizole and the terfenadine study group. However, no statistically significant inter-group differences could be detected. PMID- 11735352 TI - Inhibition of diazepam on morphine-induced hyperactivity, reverse tolerance and postsynaptic dopamine receptor supersensitivity. AB - The effects of diazepam on the development of reverse tolerance and postsynaptic dopamine receptor supersensitivity induced by morphine were examined in mice. A single administration of morphine induced hyperactivity and the morphine-induced hyperactivity was inhibited dose-dependently by the administration of diazepam (1, 2 and 4 mg x kg(-1), i.p.), an agonist for benzodiazepine receptor linked to the GABA(A) receptor. Daily repeated administration of morphine developed reverse tolerance to the hyperactivity of morphine. The concomitant administration of diazepam inhibited the morphine-induced hyperactivity and the diazepam administration prior to and during the chronic administration of morphine in mice inhibited the development of reverse tolerance to the hyperactivity of morphine (10 mg x kg(-1), s.c.). Postsynaptic dopamine receptor supersensitivity was also developed in reverse tolerant mice that had received the same morphine. The development of postsynaptic dopamine receptor supersensitivity was evidenced by the enhanced ambulatory activity of apomorphine (2 mg x kg(-1), s.c.). Diazepam also inhibited the development of postsynaptic dopamine receptor supersensitivity induced by the chronic administration of morphine. These results suggest that the hyperactivity, reverse tolerance and postsynaptic dopamine receptor supersensitivity induced by morphine may be modulated via the activation of the GABA(A) receptor induced by diazepam. PMID- 11735353 TI - Inhibitory effects of MK-801 on contextual sensitization to climbing behavior and on development of tolerance to hypothermia induced by a single high dose of apomorphine. AB - A single high dose of apomorphine (10 mg x kg(-1)) produced not only contextual sensitization to and conditioning of climbing behavior, but also context independent tolerance to hypothermia. MK-801 (0.15 and 0.3 mg x kg(-1)) inhibited contextual sensitization to and conditioning of climbing behavior. Development of tolerance to hypothermia was also inhibited by MK-801. Dopamine D1 antagonist, SCH23390 (0.5 mg x kg(-1)), but not D2 antagonist, sulpiride, inhibited sensitization to and conditioning of climbing behavior. D2 antagonist, sulpiride (50 mg x kg(-1)), but not D1 antagonist, SCH23390, inhibited development of tolerance to hypothermia. These results suggest that MK-801 inhibited contextual sensitization to climbing behavior and development of tolerance to hypothermia through glutamatergic modulation of dopaminergic functions at dopamine receptors. PMID- 11735354 TI - Modulation of pressor response to muscle contraction via monoamines following AMPA-receptor blockade in the ventrolateral medulla. AB - We hypothesized that cardiovascular responses to static muscle contraction are mediated via changes in extracellular concentrations of monoamines (norepinephrine, dopamine and serotonin) following the administration of 6-cyano 7-nitroquinoxaline-2,3-dione (CNQX, an AMPA-receptor antagonist) into the rostral (RVLM) or caudal (CVLM) ventrolateral medulla. For the RVLM experiments (n= 8), a 2-min static muscle contraction increased the mean arterial pressure (MAP) and heart rate (HR) by 23 +/- 2 mmHg and 28 +/- 8 bpm, respectively. During this contraction, the concentrations of norepinephrine, dopamine, and serotonin within the RVLM increased by 278 +/- 52%, 213 +/- 23%, and 232 +/- 24%, respectively. Microdialysis of CNQX (1.0 microM) for 30 min into the RVLM attenuated the increases in MAP and HR ( 11 +/- 2 mmHg and 14 +/- 5 bpm) without a change in developed muscle tension. The levels of norepinephrine, dopamine, and serotonin within the RVLM were also attenuated. In contrast, microdialysis of CNQX into the CVLM (n= 8) potentiated the contraction-evoked responses in MAP ( 21 +/- 2 vs 33 +/- 5 mmHg) and HR ( 25 +/- 5 vs 46 +/- 8 bpm) without any effect on the monoamine levels within the CVLM region. These results suggest that AMPA-receptor blockade within the RVLM and CVLM has opposing effects on cardiovascular responses during static muscle contraction. In addition, such receptor blockade modulates extracellular concentrations of monoamines within the RVLM but not in the CVLM. These results provide evidence that AMPA receptors within the ventrolateral medulla play a role in exercise pressor reflex. PMID- 11735355 TI - In vitro antioxidant properties of dantrolene sodium. AB - Dantrolene sodium is a skeletal muscle relaxant, which inhibits intracellular Ca2+ release from the sarcoplasmic reticulum. The aim of this study is to examine possible in vitro antioxidant effects of dantrolene sodium. For this reason, the in vitro antioxidant effects of dantrolene sodium were studied using thiocyanate methods. Additionally, the reducing power and free radical scavenging activity were determined. Dantrolene sodium showed strong antioxidant activity in the linoleic acid emulsion system. The antioxidant activity increased with an increasing amount of dantrolene sodium (50, 100, 250 microg). The 50, 100 and 250 microg samples of dantrolene sodium showed 55%, 70% and 82% inhibition on peroxidation of linoleic acid, respectively. On the other hand, the 250 microg sample of alpha-tocopherol showed 62% inhibition of peroxidation of linoleic acid. Like antioxidant activity, the reducing power of dantrolene sodium increased in a dose-dependent manner. The reducing power of dantrolene was statistically significant vs control, but lower than butylated hydroxytoluene (BHT) and quercetin. Although dantrolene sodium had free radical scavenging activity this was not statistically significant. In contrast to dantrolene sodium, quercetin and butylated hydroxyanisole (BHA) had highly potent free radical scavenging activities and those were statistically significant. According to the these results, it may be said that antioxidant effect of dantrolene sodium is more related to its antioxidant activity in linoleic acid emulsion and reducing power, than to its free radical scavenging activity. These properties may be major reasons for the inhibition of lipid peroxidation. PMID- 11735356 TI - Effect of bromazepam on stress-induced gastric ulcer in rats and its relation to brain neurotransmitters. AB - The possible antiulcer potential of bromazepam was investigated in relation to its effect on the levels of central neurotransmitters in rats. Peptic ulcer was induced by cold-restraint stress, by immobilizing the animals in open wire restraint cages placed for 2 h at 4 degrees C. Bromazepam (1 and 2 mg x kg(-1), i.p.) was given as prophylactic regimens, either as a single (2 h before ulcer induction) or repeated (twice daily for 15 days) administration. Results revealed that single (1 mg x kg(-1)) and repeated (1 and 2 mg x kg(-1)) dose regimens of bromazepam succeeded in preventing gastric ulceration, without significant effects on the protein-bound hexose content of gastric mucus. Increases in gamma aminobutyric acid (GABA) concentrations in almost all tested brain regions were observed in bromazepam-treated groups, as compared to the control stressed group. Cortical dopamine (D) concentrations were reduced following single (2 mg x kg( 1)) as well as repeated administration of bromazepam. Similarly, norepinephrine (NE) concentrations were decreased in the cerebral cortex and thalamus/hypothalamus by repeated doses of bromazepam. Cortical 5 hydroxytryptamine (5-HT) was elevated by single (1 mg x kg(-1)) and repeated (1 mg x kg(-1)) doses of the drug. It could be concluded that bromazepam affords a good gastroprotective potential against cold-restraint stress-induced gastric ulceration and the possible mechanisms might involve an increase in the inhibitory GABA and a suppression of the stimulatory NE and D in central regions, especially the cerebral cortex and/or thalamus/hypothalamus. PMID- 11735357 TI - The antinociceptive effect of moclobemide on the vocalization threshold to paw pressure in a rat model of unilateral mononeuropathy. AB - The aim of our study was to investigate the antinociceptive activity of moclobemide on the vocalization threshold to paw pressure in a rat model of unilateral mononeuropathy. The neuropathy was produced by ligation of the sciatic nerve and nociceptive thresholds were determined 15-21 days after surgery by a modification of the Randall-Sellito method. Group 1 (n= 10) received 0.2 ml peroral (p.o.) saline, Group 2 (n= 10) 5 mg x kg(-1), Group 3 (n= 10) 10 mg x kg( 1) and Group 4 (n= 10) 20 mg x kg(-1) p.o. moclobemide. Nociceptive pressure thresholds were then measured every 20 minutes after drug administration. Analysis of variance, Tukey's test and a paired Student's t-test were employed for statistical analysis. The perorally administered moclobemide (5, 10 and 20 mg x kg(-1)) produced an antinociceptive effect on both lesioned and non-lesioned hind paws ( P< 0.05). However, the analgesic effect on the lesioned paw was significantly more potent than the non-lesioned paw. The peak value ( p) remained constant while the maximal increment between the control threshold and the peak value ( I(max)) was significantly more pronounced for the lesioned paw ( P< 0.001). The results of this study may suggest that moclobemide can be a therapeutic alternative to treat some clinical symptoms in peripheral neuropathic conditions. PMID- 11735358 TI - Intravenous midazolam significantly enhances the lethal effect of thiopental but not that of ketamine in mice. AB - Intravenous (i.v.) drug combinations are used in clinical anaesthesia in order to combine the desired effects and minimize toxicity from large doses of single agents. This fundamental assumption has not been systematically evaluated. We examined its validity by testing the influence of midazolam on the lethal effect of i.v. thiopental and ketamine in mice. Dose-response curves were constructed for the lethal effect of i.v. thiopental and ketamine, and for the loss of righting reflex effect by midazolam, in sexually mature male ICR mice weighing 20 40 g. For each curve, six or seven groups of eight to 10 mice each were used. A quarter of the median effective dose (ED50) for loss of righting reflex by midazolam was combined with the two other drugs to deduce dose-response curves for the lethal effect of the combinations. The ED50 for loss of righting reflex by i.v. midazolam was 43.5 mg x kg(-1) (95% confidence interval [CI], 40.4-46.5). The median lethal dose (LD 50) of i.v. thiopental was 50.6 mg x kg(-1) (95% CI, 50.0-54.9) and that of ketamine 42.9 mg x kg(-1) (95% CI, 32.3-52). In the presence of 10 mg x kg(-1) midazolam, the LD50 of thiopental was reduced to 20 mg x kg(-1) (17.7-22.2), but that of ketamine remained 44.4 mg x kg(-1) (37.7-54.9). Midazolam increased the lethal effect of thiopental 2.5-fold, but did not affect that of ketamine. Interactions at the toxic level between commonly used anaesthetic agents may differ from those at the hypnotic or analgesic levels, which should prompt evaluation of such combinations before their introduction to routine clinical use. PMID- 11735359 TI - Size effects of realgar particles on apoptosis in a human umbilical vein endothelial cell line: ECV-304. AB - Regulation of endothelial cell apoptosis is a critical modulator of normal and pathological angiogenesis. Inorganic arsenic, especially arsenic trioxide, has been demonstrated to induce clinical remission in patients with acute promyelocytic leukemia as well as those with several solid cancers. So, in this study, we examined the in vitro effect of realgar(As2S2), used to treat human diseases for centuries in traditional Chinese medicine, on cell survival and apoptosis in a human umbilical vein endothelial cell (HUVEC) line: ECV-304. Four different suspensions of realgar particles with diameters from 100-500 nm containing equivalent doses were investigated to determine their size effects. After 2, 4, 6 and 8 h, realgar particles of diameters 100 nm and 150 nm obviously reduced ECV-304 cell survival, which was evidenced by a reduction of cell viability in the MTT assay, whereas cells exhibited slight viability reduction in response to treatment with realgar particles of diameters 200 nm and 500 nm. Significant apoptosis was identified by morphological observation, flow cytometry assay and DNA gel electrophoresis in the ECV-304 cells by the presence of realgar particles of diameters 100 nm and 150 nm. It is demonstrated that particles of realgar with average diameters from 100 nm to 150 nm can induce ECV-304 cell apoptosis. These results suggest that particles of realgar smaller than 150 nm may provide a less toxic agent for anti-neoplasia by suppressing angiogenesis. PMID- 11735360 TI - Time-dependent reduction of acetylcholine-induced relaxation in aortic rings of cholestatic rats. AB - Changes in vascular responsiveness are the basis for some of the cardiovascular complications in cholestasis. Since the duration of cholestasis is important in determining the degree of the complications, we investigated the time-course dependent evolution of vascular relaxation responsiveness in the aortic rings of cholestatic rats. Acetylcholine-induced endothelium-dependent relaxation was investigated in the isolated aortic rings of unoperated, sham-operated and two-, five-, seven- and fourteen-day bile-duct ligated rats. There was a significant reduction in acetylcholine-induced relaxation of the aortic rings by the second day after the bile-duct ligation operation, compared to those of unoperated and sham-operated groups, but more reduction still occurs in 5- and 7-day bile-duct ligated groups, reaching a plateau by the seventh day. The relaxation response to sodium nitroprusside in the aortic rings of the unoperated and the 7-day bile duct ligated rats did not differ, implying the intact smooth muscle component of the relaxation pathway. L-NAME ( N(omega)-nitro-L-arginine methyl ester), a nitric oxide (NO) synthase inhibitor, attenuated the acetylcholine-induced relaxation in both groups (unoperated and bile-duct ligated), while L-arginine prevents this inhibitory effect. Indomethacin potentiated the acetylcholine induced relaxation in the aortic rings of the bile-duct ligated rats while it has no effect on unoperated controls, providing evidence for the possible role of vasoconstrictor prostanoids in cholestasis-induced reduction in acetylcholine induced relaxation. These results state that the reduced acetylcholine-induced relaxation in the cholestatic aortic rings during the first week, when no portal hypertension was reported to be present, may be due to the decreased acetylcholine-induced NO release from endothelium or increased NO inactivation. PMID- 11735361 TI - Protective effect of T-type calcium channel blocker in histamine-induced paw inflammation in rat. AB - The aim of this paper was to investigate the protective effect of the T-type calcium channel blocker in a model of acute local inflammation (histamine induced). The intraplantar injection of histamine elicited an inflammatory response that was characterized by a time-dependent increase in paw oedema and neutrophil infiltration in paw tissue. The maximal increase in paw volume was observed at 90 min after histamine administration (maximal paw volume: 0.97 +/- 0.07). In addition, polymorphonuclear leucocyte (PNL) number was markedly increased in the histamine-treated paw tissue (144 +/- 25.56). However, histamine induced paw oedema was significantly reduced in a dose-dependent manner by treatment with mibefradil (given at 10, 25, 50, and 100 mg x kg(-1)) at 30, 60, 90, 120, 150, 180 min after injection of histamine. Mibefradil treatment also caused a significant reduction of the polymorphonuclear leucocyte number in the paw tissue. Our findings support the view that mibefradil exerts antiinflammatory effects. PMID- 11735362 TI - Role of nitric oxide in catalepsy and hyperthermia in morphine-dependent rats. AB - The possible involvement of nitric oxide (NO) in morphine-induced catalepsy and hyperthermia was studied in morphine-dependent rats. Four days repeated injection regimen was used to induce morphine dependence, which was assessed by naloxone challenge (0.5 mg x kg(-1), s.c.). Pretreatment of rats with the NO synthase inhibitor, N(G)-nitro-L-arginine (L-NA, 8 mg x kg(-1) twice daily, i.p.) potentiated the cataleptic response of morphine as shown by a rightward shift in the morphine-log dose-response curve. Prior treatment of rats with the NO precursor, L-arginine (200 mg x kg(-1), twice daily, i.p.) abolished the potent effect of L-NA and restored the cataleptic scores to levels similar to those of morphine-dependent rats. The same dose of L-NA significantly blocked morphine induced hyperthermia at the dose levels of morphine (15-105 mg x kg(-1)) and this effect was reversed by L-arginine. These data provide the first experimental evidence that NO is involved in morphine induced catalepsy and hyperthermia and demonstrated that blockade of NO synthesis may suggest a dangerous interaction with opioids in the control of motor function. PMID- 11735363 TI - Nefopam reduces thermal hypersensitivity in acute and postoperative pain models in the rat. AB - The activity of nefopam, a centrally acting compound, not structurally related to other analgesics, was examined in acute and postoperative thermal pain models in the rat. Its antinociceptive potency was evaluated using heat noxious stimuli either in intact or in injured animals after skin and muscular incisions. In the hot plate and in the plantar tests, nefopam after acute administration by different routes exhibited a dose-dependent attenuation of the nociceptive responses at 10-30 mg x kg(-1) by intraperitoneal or subcutaneous administration, at 60 mg x kg(-1) by oral dosing, and from 3 mg x kg(-1) after intravenous injection. In the postoperative pain model, at 30 mg x kg(-1) nefopam augmented the endpoint to thermal threshold, 60 and 90 minutes after administration compared to the threshold recorded after the incision. In the same conditions, morphine and tramadol displayed antinociceptive activities. As the plantar test provides a good index of nociception in humans, these results point out the usefulness of nefopam for attenuating moderate to severe pain, and for postoperative analgesia. In conclusion, nefopam has shown potent properties to reduce thermal hypersensitivity after acute or postoperative pain in rats. PMID- 11735364 TI - Mechanism underlying histamine-induced intracellular Ca2+ movement in PC3 human prostate cancer cells. AB - The effect of histamine on intracellular free Ca2+ levels ([Ca2+]i) in PC3 human prostate cancer cells and the underlying mechanism were evaluated using fura-2 as a Ca2+ dye. Histamine at concentrations between 0.1 and 50 microM increased [Ca2+]i in a concentration-dependent manner with an EC50 value of 1 microM. The [Ca2+]i response comprised an initial rise and a slow decay, which returned to baseline within 3 min. Extracellular Ca2+ removal inhibited 50% of the [Ca2+]i signal. In the absence of extracellular Ca2+, after cells were treated with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor), 10 microM histamine did not increase [Ca2+]i. After pretreatment with 10 microM histamine in a Ca2+-free medium for several minutes, addition of 3 mM Ca2+ induced [Ca2+]i increases. Histamine (10 microM)-induced intracellular Ca2+ release was abolished by inhibiting phospholipase C with 2 microM 1-(6-((17 beta-3- methoxyestra 1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122), and by 10 microM pyrilamine but was not altered by 50 microM cimetidine. Collectively, the present study shows that histamine induced [Ca2+]i transients in PC3 human prostate cancer cells by stimulating H1 histamine receptors leading to Ca2+ release from the endoplasmic reticulum in an inositol 1,4,5-trisphosphate dependent manner, and by inducing Ca2+ entry. PMID- 11735365 TI - Identification of a novel gene involved in stable maintenance of plasmid pGA1 from Corynebacterium glutamicum. AB - The cryptic plasmid pGA1 (4.8 kb) from Corynebacterium glutamicum, replicating in the rolling-circle mode, has been reported to contain four open reading frames longer than 200 bp (ORFA/per, ORFA2, ORFB, ORFC/rep). Here we present another pGA1 gene, ORFE (174 bp), located in the region downstream of the per-ORFA2 gene cluster. The ORFE is transcribed into two RNA species in a direction opposite to that of the per-ORFA2 RNA. Introduction of ORFE in trans into the cells harboring the pGA1 derivatives carrying the main stability determinant, the per gene coding for a product that positively influences the pGA1 copy number and maintenance, increased their segregational stability. Mutation of the putative translational start of the ORFE abolished this observed positive effect in trans. ORFE thus codes for a protein acting as an accessory element involved in stable maintenance of plasmid pGA1 and was hence designated the aes gene (accessory effector of stable maintenance). PMID- 11735366 TI - Efficient transfection of novel bovine papillomavirus 1 expression plasmids. AB - A series of novel bovine papillomavirus type 1 (BPV-1)-based expression plasmids was constructed and characterized in vitro as a starting point for the development of an in vivo gene therapeutic method. The order of transfection efficiency for different pBPVlacZ plasmids was pCGalBPV > pTKBPV > pSRalphaBPV in CV1-P cells. In the absence of selection pressure, the expression of pCGalBPVlacZ and pTKBPVlacZ was associated with long-term maintenance. In a comparison of pBPVlacZ with pSVlacZ, expression was maintained up to 12-17 and 8-12 days, respectively. The transfection of pBPVlacZ plasmids was efficient in secondary and primary, dividing and nondividing, neural and nonneural, and human cells and, furthermore, independent of the cell cycle as seen in growing as well as resting cells. All these characteristics are likely to be relevant for in vivo conditions, under which the percentage of proliferating cells could be quite low. In conclusion, the pBPV plasmids were efficiently delivered and expressed in different host cells, and therefore their performance in gene therapy is worth testing. PMID- 11735367 TI - Sequence of the 50-kb conjugative multiresistance plasmid pRE25 from Enterococcus faecalis RE25. AB - The complete 50,237-bp DNA sequence of the conjugative and mobilizing multiresistance plasmid pRE25 from Enterococcus faecalis RE25 was determined. The plasmid had 58 putative open reading frames, 5 of which encode resistance to 12 antimicrobials. Chloramphenicol acetyltransferase and the 23S RNA methylase are identical to gene products of the broad-host-range plasmid pIP501 from Streptococcus agalactiae. In addition, a 30.5-kb segment is almost identical to pIP501. Genes encoding an aminoglycoside 6-adenylyltransferase, a streptothricin acetyltransferase, and an aminoglycoside phosphotransferase are arranged in tandem on a 7.4-kb fragment as previously reported in Tn5405 from Staphylococcus aureus and in pJH1 from E. faecalis. One interrupted and five complete IS elements as well as three replication genes were also identified. pRE25 was transferred by conjugation to E. faecalis, Listeria innocua, and Lactococcus lactis by means of a transfer region that appears similar to that of pIP501. It is concluded that pRE25 may contribute to the further spread of antibiotic resistant microorganisms via food into the human community. PMID- 11735368 TI - DNA cloning in Lactobacillus helveticus by the exconjugation of recombinant mob containing plasmid constructs from strains of transformable lactic acid bacteria. AB - A system developed for the genetic transfer of plasmids between strains of nontransformable bacteria (P. Langella, Y. le Loir, S. D. Ehrlich and A. Gruss, 1993, J. Bacteriol., 175, 5806-5813) by the specific inclusion of a mobilization (mob) region into a nonconjugative shuttle vector was used successfully to deliver the genetic determinants for beta-glucanase, beta-glucuronidase, and green fluorescent protein to Lactobacillus helveticus. Expression of two of the genes could be detected in the new host. Data suggested that resolution of cointegrates into components could release the original recombinant plasmid or generate a cointegrate deletion. All the recombinant plasmids were segregationally unstable in Lb. helveticus and there was some evidence for structural instability. Intrinsic instability in the mob-containing vector was reduced by replacing the duplicated pBluescript polylinker with that from pUC19. Sites at which cointegrate formation could occur were localized at two distinct tracts close to the D-loop that forms at the primosome during plasmid replication. PMID- 11735369 TI - Transfer region of a Bacteroides conjugative transposon contains regulatory as well as structural genes. AB - Conjugative transposons (CTns) are integrated elements that excise themselves from the chromosome to form a circular transfer intermediate that is transferred by conjugation to a recipient. In an earlier paper, the excision step was shown to be regulated by tetracycline and to be dependent on the regulatory gene, rteC. In this paper, we report that genes involved in conjugal transfer are also regulated by tetracycline but that regulation is more complex. Genes contained within a 20-kbp region that is sufficient for conjugal transfer were disrupted by single crossover integration events. Most of the disruptions abolished transfer of the CTn. None of them abolished excision. Antibodies to two of the proteins encoded in this region (TraG and TraN) were obtained and used to show that production of these proteins was dependent on tetracycline stimulation. Both TraG and TraN were membrane proteins. A surprising finding was that a disruption in the gene traQ increased transfer of CTnERL over 100-fold. Thus, TraQ may be a repressor protein that controls expression of transfer genes. If so, TraQ is not the only protein that controls expression of transfer genes because production of TraG and TraN in the traQ disruption mutant was still dependent on tetracycline stimulation. PMID- 11735370 TI - Genetic organization of the Francisella plasmid pFNL10. AB - We report here the molecular characterization of pFNL10, a 3990-bp cryptic plasmid of Francisella novicida-like F6168. The plasmid was maintained in F. novicida Utah 112 and F. tularensis LVS strains. We sequenced the entire plasmid and found six open reading frames (ORFs)-ORF1, ORF2, ORF3, ORF4, ORF5, and ORFm. ORF3, ORF4, ORF5, and ORFm are located on the same strand, and we designated it the plus strand. ORF1 and ORF2 are on the complementary strand. The ORFs appear to be arranged in two operons, one comprising ORF5 and ORF4 and the other ORF1 and ORF2. There exist two distinct promoters similar to the Escherichia coli sigma(70) promoter, one 5' to ORF1-ORF2 operon and the other 5' to ORF5-ORF4 operon. We found that in both promoters the transcriptional start is an adenosine. ORF3 is positioned in tandem with ORF5-ORF4, but has its own transcriptional start, a thymidine. However, sequence analysis revealed no recognizable promoter in physical proximity to ORF3. Sequence analysis revealed transcriptional terminators immediately downstream of the two operons. Experimental results showed that the ORF1-ORF2 terminator is authentic. But we could not definitively confirm the ORF5-ORF4 terminator. Two sets of direct repeats, one 31 and the other 13 bp, characteristic of ori are positioned between the two promoters. ORF1 encodes a protein that bears homology to the replication initiation protein RepA of various bacteria, and disruption of this ORF indeed blocked pFNL10 replication. In contrast, ORF2 disruption caused formation of plasmid multimers, suggesting aberrant replication. Our analysis also suggests that pFNL10 replicates by the theta mode. The ORF5-ORF4 operon resembles the phd doc operon of Escherichia coli bacteriophage P1, but the significance of this similarity is unclear. PMID- 11735371 TI - Sequence and gene expression analyses of plasmid pHPM8 from Helicobacter pylori reveal the presence of two operons with putative roles in plasmid replication and antibiotic activity. AB - The DNA sequence of a 7.8-kb Helicobacter pylori plasmid, pHPM8, was determined. Six open reading frames (ORFs) were present. Ribonuclease protection studies showed that ORF1/ORF2 and ORF3/ORF4 genes are organized in operons possibly involved in plasmid replication and in production of a peptide with antibiotic activity, respectively. Finding areas of pHPM8 with a high level of identity to H. pylori chromosomal DNA supported the hypothesis that recombination occurs between plasmids and the chromosome of H. pylori. PMID- 11735372 TI - Induction of pCW3-encoded tetracycline resistance in Clostridium perfringens involves a host-encoded factor. AB - The tetracycline resistance determinant Tet P, which is encoded by the conjugative plasmid pCW3 from Clostridium perfringens, is induced by subinhibitory concentrations of tetracycline. In this study we have shown that the inducible phenotype is strain dependent. When pCW3 is present in derivatives of the wild-type strains CW234 and CW362 resistance is inducible. However, transfer to derivatives of strain 13 leads to a constitutive phenotype that is only observed in this strain background. Based on these results it is proposed that induction of the pCW3-encoded tet(P) genes in C. perfringens requires a host encoded factor that is either absent or nonfunctional in strain 13 derivatives. PMID- 11735374 TI - Lessons from mitochondrial DNA mutations. AB - The small, maternally inherited mitochondrial DNA (mtDNA) has turned out to be a hotbed of pathogenic mutations: 13 years into the era of "mitochondrial medicine", over 100 pathogenic point mutations and countless rearrangements have been associated with a variety of multisystemic or tissue-specific human diseases. MtDNA-related disorders can be divided into two major groups: those due to mutations in genes affecting mitochondrial protein synthesis in toto and those due to mutations in specific protein-coding genes. Pathogenesis is only partially explained by the rules of mitochondrial genetics and remains largely uncharted territory. Therapy is still woefully inadequate, but a number of promising approaches are being developed. PMID- 11735375 TI - The expanding spectrum of nuclear gene mutations in mitochondrial disorders. AB - Our understanding of the molecular basis of mitochondrial disorders has come primarily from the discovery of an expanding number of mutations of mtDNA. However, a variety of recent observations indicate that many syndromes are due to abnormalities in nuclear genes related to oxidative-phosphorylation (OXPHOS). Nuclear genes encode hundreds of proteins involved in mitochondrial OXPHOS. Nevertheless, the identification of these genes has proceeded at a much slower pace, compared with the discovery and characterization of mtDNA mutations. This scenario is rapidly changing, thanks to the discovery of several OXPHOS-related human genes, and to the identification of mutations responsible for different clinical syndromes. PMID- 11735376 TI - Defects of intergenomic communication: autosomal disorders that cause multiple deletions and depletion of mitochondrial DNA. AB - Depletion and multiple deletions of mitochondrial DNA (mtDNA) have been associated with a growing number of autosomal diseases that have been classified as defects of intergenomic communication. MNGIE, an autosomal recessive disorder associated with mtDNA alterations is due to mutations in thymidine phosphorylase that may cause imbalance of the mitochondrial nucleotide pool. Subsequently, mutations in the mitochondrial proteins adenine nucleotide translocator 1, Twinkle, and polymerase gamma have been found to cause autosomal dominant progressive external ophthalmoplegia with multiple deletions of mtDNA. Uncovering the molecular bases of intergenomic communication defects will enhance our understanding of the mechanisms responsible for maintaining mtDNA integrity. PMID- 11735377 TI - Neuropathological features of mitochondrial disorders. AB - Genetic defects affecting the mitochondrial respiratory chain comprise an important cause of encephalomyopathies. Considering the structural complexity of the respiratory chain, its dual genetic control, and the numerous nuclear genes required for proper assembly of the enzyme complexes, the phenotypic heterogeneity is not surprising. From a neuropathological view point, application of in situ hybridization and immunohistochemistry to study the choroid plexus and brain-blood barrier in "prototypes" of mitochondrial encephalopathies have revealed alterations that we think are important in the pathogenesis of central nervous system dysfunction in these disorders. As the role of the blood cerebrospinal fluid (CSF) and brain-blood barriers in mitochondrial encephalopathies is better understood, manipulation of their functions offers promises for therapeutic interventions. PMID- 11735378 TI - Pathogenesis of primary defects in mitochondrial ATP synthesis. AB - Maternally inherited mutations in the mtDNA-encoded ATPase 6 subunit of complex V (ATP synthase) of the respiratory chain/oxidative phosphorylation system are responsible for a subgroup of severe and often-fatal disorders characterized predominantly by lesions in the brain, particularly in the striatum. These include NARP (neuropathy, ataxia, and retinitis pigmentosa), MILS (maternally inherited Leigh syndrome), and FBSN (familial bilateral striatal necrosis). Of the five known pathogenic mutations causing these disorders, four are located at two codons (156 and 217), each of which can suffer mutations converting a conserved leucine to either an arginine or a proline. Based on the accumulating data on both the structure of ATP synthase and the mechanism by which rotary catalysis couples proton flow to ATP synthesis, we propose a model that may help explain why mutations at codons 156 and 217 are pathogenic. PMID- 11735379 TI - Reactive oxygen species and mitochondrial diseases. AB - A variety of diseases have been associated with excessive reactive oxygen species (ROS), which are produced mostly in the mitochondria as byproducts of normal cell respiration. The interrelationship between ROS and mitochondria suggests shared pathogenic mechanisms in mitochondrial and ROS-related diseases. Defects in oxidative phosphorylation can increase ROS production, whereas ROS-mediated damage to biomolecules can have direct effects on the components of the electron transport system. Here, we review the molecular mechanisms of ROS production and damage, as well as the existing evidence of mitochondrial ROS involvement in human diseases. PMID- 11735380 TI - Mito-mice: animal models for mitochondrial DNA-based diseases. AB - We have successfully produced "Mito-mice" harbouring a pathogenic mtDNA mutation. We generated the mice by introducing mitochondria with a 4696 base-pair mtDNA deletion (Delta mtDNA4696) into mouse embryos. This deletion encompasses nucleotides 7759-12 454 and includes six tRNA genes and seven structural genes. In Mito-mice, the Delta mtDNA4696 is transmitted maternally, and induces mitochondrial dysfunction in various tissues. Most of the Mito-mice with high proportions of the Delta mtDNA4696 died at about age 6 months due to renal failure. Mito-mice are the first animal model for mtDNA-based diseases and will be valuable for studying pathogenesis and for identifying effective drug and gene therapies. PMID- 11735382 TI - Eye specification in Drosophila: perspectives and implications. AB - The discovery that Drosophila eyeless is homologous to vertebrate Pax6 produced enormous interest in eye specification and a reappraisal of eye evolution. While the transcription factor Eyeless/Pax6 is necessary and in some circumstances sufficient to induce eye development the simple story of eye specification has become more epic than haiku. At least seven other nuclear proteins act with Eyeless/Pax6 to induce the eye and, furthermore, extrinsic developmental signals are required. Some striking similarities between later events of retinal patterning in vertebrates and insects have led to a deeper debate on the evolutionary path to these apparently quite different organs. PMID- 11735383 TI - Mammalian homologues of the Drosophila eye specification genes. AB - The Drosophila compound eye is specified by the simultaneous and interdependent activity of transcriptional regulatory genes from four families: PAX6 (eyeless, twin of eyeless, eyegone), EYA (eyes absent), SIX (sine oculis, Optix) and DACH (dachshund). Mammals have homologues of all these genes, and many of them are expressed in the embryonic or adult eye, but the functional relationships between them are currently much less clear than in Drosophila. Nevertheless, mutations in the mammalian genes highlight their requirement both within and outside the eye in embryos and adults, and emphasize that they can be deployed in many different contexts. PMID- 11735384 TI - Pattern formation in the zebrafish retina. AB - During the past 15 years, the zebrafish has become established as a genetic model organism to study vertebrate development. It is particularly well suited for the analysis of the retina, and several genetic screens have yielded a large number of mutants affecting retinal development. Most of these mutants still await thorough analysis and molecular characterization, but work on a handful of genes has already generated interesting results that shed some light on patterning mechanisms employed in the vertebrate retina. PMID- 11735385 TI - The role of basic helix-loop-helix genes in vertebrate retinogenesis. AB - The developing eye is a favorite model for the study of pattern formation and cell fate determination. Retinal neuron development, in particular, is an approachable system to study molecular and cellular aspects of cell determination and differentiation. Basic helix-loop-helix (bHLH) transcription factors are important regulators of retinal neurogenesis. Proneural bHLH genes have highly defined expression in the developing retina that are influenced by pattern formation and cell specification pathways. Each retinal cell class has unique bHLH requirements, implying that these genes regulate neuronal identity and function. Therefore, proneural genes represent a molecular focal point through which epithelial cells are transformed into a precise neural network. In this review, we focus on the bHLH factor Ath5, an important regulator of retinal ganglion cell development, and discuss factors that regulate its expression in the retina and the target genes through which it may confer specific neuronal properties. PMID- 11735386 TI - Cell proliferation, survival, and death in the Drosophila eye. AB - The Drosophila retina has a precise repeating structure based on the unit eye, or ommatidium. This review summarizes studies of the cell proliferation and survival episodes that affect the number of cells available to make each ommatidium. Late in larval development, as differentiation and patterning begin, the retinal epithelium exhibits striking regulation of the cell cycle including a transient G1 arrest of all cells, followed by a "Second Mitotic Wave" cell cycle that is regulated at the G2/M transition by local intercellular signals. Reiterated episodes of cell death also contribute to precise regulation of retinal cell number. The EGF receptor homolog has multiple roles in retinal proliferation and survival. PMID- 11735387 TI - Photoreceptor subtype specification: from flies to humans. AB - Multiple cell types often differentiate from a pluripotent cell. These cells may then further diversify as distinct subtypes. The visual system provides an ideal model for studying subtype specification as various photoreceptors acquire different functions based on the type of opsin they express. Opsin expression is mostly controlled through transcriptional mechanisms that are evolutionary conserved from Drosophila to humans. In addition, it appears that, from a "default" developmental state, distinct "acquired" photoreceptor states develop upon receiving intrinsic or extrinsic signals. This review discusses factors involved in opsin gene regulation and how their integration may explain how subtype specificity is achieved. PMID- 11735388 TI - Dynamics of excitation energy transfer in the LH1 and LH2 light-harvesting complexes of photosynthetic bacteria. AB - Photosynthetic light harvesting is a unique life process that occurs with amazing efficiency. Since the discovery of the structure of the bacterial peripheral light-harvesting complex (LH2), this process has been studied using a variety of advanced laser spectroscopic methods. We are now in a position to discuss the physical origins of excitation energy transfer and trapping in the LH2 and LH1 antennae of photosynthetic purple bacteria. We demonstrate that the time evolution of the state created by the light is determined by the combined action of excitonic pigment-pitment interactions, energetic disorder, and coupling to nuclear motion in a pigment-protein complex. A quantitative fit of experimental data using Redfield theory allowed us to determine the pathways and time scales of exciton and vibrational relaxation and analyze separately different contributions to the measured transient absorption dynamics. Furthermore, these dynamics were observed to be strongly dependent on the excitation wavelength. A numerical fit of this dependence turns out to be extremely critical to a variation of the structure and disorder parameters and, therefore, can be used as a test for different antenna models (disordered ring, elliptical deformations, correlated disorder, etc.). The calculated equilibration dynamics in the exciton basis allow a visualization of the exciton motion using a density matrix picture in real space. PMID- 11735389 TI - The affinity-enhancing roles of flexible linkers in two-domain DNA-binding proteins. AB - Recently many attempts have been made to design high-affinity DNA-binding proteins by linking two domains. Here a theory for guiding these designs is presented. Flexible linkers may play three types of roles: (a) linking domains which by themselves are unfolded and bind to DNA only as a folded dimer (as in a designed single-chain Arc repressor), (b) connecting domains which can separately bind to DNA (as in the Oct-1 POU domain), and (c) linking a DNA-binding domain with a dimerization domain (as in the lambda repressor). In (a), the linker keeps the protein as a folded dimer so that it is always DNA-binding-competent. In (b), the linker is predicted to enhance DNA-binding affinity over those of the individual domains (with dissociation constants K(A) and K(B)) by p(d(0))/K(B) or p(d(0))/K(A), where p(d(0)) = (3/4pil(p)bL)(3/2) exp(-3d(0)(2)/4l(p)bL)(1 - 5l(p)/4bL +...) is the probability density for the end-to-end vector of the linker with L residues to have a distance d(0). In (c), the linker is predicted to enhance the binding affinity by K(d)(C)/p(d(0)), where K(d)(C) is the dimer dissociation constant for the dimerization domain. The predicted affinity enhancements are found to be actually reached by the Oct-1 POU domain and lambda repressor. However, there is room for improvement in many of the recently designed proteins. The theoretical limits presented should provide a useful guide for current efforts of designing DNA-binding proteins. PMID- 11735390 TI - A zinc ribbon protein in DNA replication: primer synthesis and macromolecular interactions by the bacteriophage T4 primase. AB - The gene product 61 primase protein from bacteriophage T4 was expressed as an intein fusion and purified to homogeneity. The primase binds one zinc ion, which is coordinated by four cysteine residues to form a zinc ribbon motif. Factors that influence the rate of priming were investigated, and a physiologically relevant priming rate of approximately 1 primer per second per primosome was achieved. Primase binding to the single-stranded binding protein (1 primase:4 gp32 monomers; K(d) approximately 860 nM) and to the helicase protein in the presence of DNA and ATP-gamma-S (1 primase:1 helicase monomer; K(d) approximately 100 nM) was investigated by isothermal titration calorimetry (ITC). Because the helicase is hexameric, the inferred stoichiometry of primase binding as part of the primosome is helicase hexamer:primase in a ratio of 1:6, suggesting that the active primase, like the helicase, might have a ring-like structure. The primase is a monomer in solution but binds to single-stranded DNA (ssDNA) primarily as a trimer (K(d) approximately 50-100 nM) as demonstrated by ITC and chemical cross linking. Magnesium is required for primase-ssDNA binding. The minimum length of ssDNA required for stable binding is 22-24 bases, although cross-linking reveals transient interactions on oligonucleotides as short as 8 bases. The association is endothermic at physiologically relevant temperatures, which suggests an overall gain in entropy upon binding. Some possible sources of this gain in entropy are discussed. PMID- 11735391 TI - Proton-collecting properties of bovine heart cytochrome C oxidase: kinetic and electrostatic analysis. AB - Proton-transfer reactions on the surface of bovine heart cytochrome c oxidase were investigated by combining a laser-induced proton-pulse technique with molecular modeling. The experimental approach simultaneously monitors the state of pyranine protonation in the bulk phase and that of a fluorescein indicator specifically attached to the native Cys(III-115) residue of subunit III of cytochrome oxidase. The reversible dynamics of the acid-base equilibration between the surface and the bulk phase were measured with submicrosecond time resolution and analyzed by numerical integration of coupled nonlinear differential rate equations. Kinetic analysis shows that carboxylates on the surface of the protein act as a proton-collecting antenna, which is able to rapidly transfer protons to nearby histidines that function as a local proton reservoir. These properties enable cytochrome oxidase to carry out its redox linked proton translocation. Molecular modeling of the fluorescein-binding site indicates that, in addition to the covalent bond, the dye is anchored through a hydrogen bond to the hydroxyl moiety of Tyr(VII-50). The protonation of the dye is mediated through three residues that shuttle protons between the bulk and the dye. A correlation between the measured kinetic properties of the bound fluorescein and the different configurations of the dye allows us to predict the identity of the proton-binding sites in the fluorescein-binding domain. PMID- 11735392 TI - Serine 85 in transmembrane helix 2 of short-wavelength visual pigments interacts with the retinylidene Schiff base counterion. AB - Short-wavelength cone visual pigments (SWS1) are responsible for detecting light from 350 to 430 nm. Models of this class of pigment suggest that TM2 has extensive contacts with the retinal binding pocket and stabilizes interhelical interactions. The role of TM2 in the structure-function of the Xenopus SWS1 (VCOP, lambda(max) = 427 nm) pigment was studied by replacement of the helix with that of bovine rhodopsin and also by mutagenesis of highly conserved residues. The TM2 chimera and G78D, F79L, M81E, P88T, V89S, and F90V mutants did not produce any significant spectral shift of the dark state or their primary photointermediate formed upon illumination at cryogenic temperatures. The mutant G77R (responsible for human tritanopia) was completely defective in folding, while C82A and F87T bound retinal at reduced levels. The position S85 was crucial for obtaining the appropriate spectroscopic properties of VCOP. S85A and S85T did not bind retinal. S85D bound retinal and had a wild-type dark state at room temperature and a red-shifted dark state at 45 K and formed an altered primary photointermediate. S85C absorbed maximally at 390 nm at neutral pH and at 365 nm at pH >7.5. The S85C dark state was red shifted by 20 nm at 45 K and formed an altered primary photointermediate. These data suggest that S85 is involved in a hydrogen bond with the protonated retinylidene Schiff base counterion in both the dark state and the primary photointermediate. PMID- 11735393 TI - Deuterium kinetic isotope effects in the p-side pathway for quinol oxidation by the cytochrome b(6)f complex. AB - Plastoquinol oxidation and proton transfer by the cytochrome b(6) f complex on the lumen side of the chloroplast thylakoid membrane are mediated by high and low potential electron transport chains. The rate constant for reduction, k(bred), of cytochrome b(6) in the low potential chain at ambient pH 7.5-8 was twice that, k(fred), of cytochrome f in the high potential chain, as previously reported. k(bred) and k(fred) have a similar pH dependence in the presence of nigericin/nonactin, decreasing by factors of 2.5 and 4, respectively, from pH 8 to an ambient pH = 6, close to the lumen pH under conditions of steady-state photosynthesis. A substantial kinetic isotope effect, k(H2O)/k(D2O), was found over the pH range 6-8 for the reduction of cytochromes b(6) and f, and for the electrochromic band shift associated with charge transfer across the b(6)f complex, showing that isotope exchange affects the pK values linked to rate limiting steps of proton transfer. The kinetic isotope effect, k(bred)(H2O)/k(bred) (D2O) approximately 3, for reduction of cytochrome b in the low potential chain was approximately constant from pH 6-8. However, the isotope effect for reduction of cytochrome f in the high potential chain undergoes a pH dependent transition below pH 6.5 and increased 2-fold in the physiological region of the lumen pH, pH 5.7-6.3, where k(fred)(H2O)/k(fred)(D2O) approximately 4. It is proposed that a rate-limiting step for proton transfer in the high potential chain resides in the conserved, buried, and extended water chain of cytochrome f, which provides the exit port for transfer of the second proton derived from p-side quinol oxidation and a "dielectric well" for charge balance. PMID- 11735394 TI - Identification of a contact domain between echistatin and the integrin alpha(v)beta(3) by photoaffinity cross-linking. AB - The integrin alpha(v)beta(3) is the major receptor mediating the attachment of osteoclasts to the extracellular matrix in bone and plays a critical role in bone resorption and bone remodeling. Most of the ligands interacting with the alpha(v)beta(3) receptor contain an Arg-Gly-Asp (RGD) motif. Recently, we have identified two small RGD peptides, containing a benzophenone moiety at either the carboxyl or amino terminus, that photo-cross-linked within the beta(3)[99-118] [Bitan, G., et al. (1999) Biochemistry 38, 3414-3420] or the beta(3)[167-171] [Bitan, G., et al. (2000) Biochemistry 39, 11014-11023] sequence, respectively, of the alpha(v)beta(3) receptor in a selective fashion. Here, we report the synthesis of a photoreactive analogue of echistatin (a 49-amino acid peptide), a potent RGD-containing antagonist of the alpha(v)beta(3) receptor both in vitro and in vivo. This bioactive analogue is substituted at position 45 with a p benzoyl moiety (pBz(2)), located within the flexible C-terminal domain and removed 20 amino acid residues from the R(24)GD(26) triad. This C-terminal domain was reported to contribute to receptor binding affinity by acting as an auxiliary binding site. The radiolabeled (125)I-[Arg(35),Lys(45)(N(epsilon)-pBz(2))] echistatin photo-cross-links effectively to a site within the beta(3)[209-220] sequence. Residues in this domain have been reported to be part of the metal ion dependent adhesion site (MIDAS). Receptor fragments overlapping this domain were reported to bind to fibrinogen and block fibrinogen binding to alpha(IIb)beta(3), the platelet integrin receptor. Taken together, position 45 in echistatin, located within an auxiliary binding site in echistatin, cross-links to a site distinct from the two previously reported sites, beta(3)[99-118] and beta(3)[167 171], which cross-link to photophores flanking the RGD triad. These cross-linking data support the hypothesis that the ligand-bound conformation of the integrin beta(3) subunit differs from the known conformation of I domains. PMID- 11735395 TI - Calcium coordination and pH dependence of the calcium affinity of ligand-binding repeat CR7 from the LRP. Comparison with related domains from the LRP and the LDL receptor. AB - We have determined the X-ray crystal structure to 1.8 A resolution of the Ca(2+) complex of complement-like repeat 7 (CR7) from the low-density lipoprotein receptor-related protein (LRP) and characterized its calcium binding properties at pH 7.4 and 5. CR7 occurs in a region of the LRP that binds to the receptor associated protein, RAP, and other protein ligands in a Ca(2+)-dependent manner. The calcium coordination is identical to that found in LB5 and consists of carboxyls from three conserved aspartates and one conserved glutamate, and the backbone carbonyls of a tryptophan and another aspartate. The overall fold of CR7 is similar to those of CR3 and CR8 from the LRP and LB5 from the LDL receptor, though the low degree of sequence homology of residues not involved in calcium coordination or in disulfide formation results in a distinct pattern of surface residues for each domain, including CR7. The thermodynamic parameters for Ca(2+) binding at both extracellular and endosomal pHs were determined by isothermal titration calorimetry for CR7 and for related complement-like repeats CR3, CR8, and LB5. Although the drop in pH resulted in a reduction in calcium affinity in each case, the changes were very variable in magnitude, being as low as a 2-fold reduction for CR3. This suggests that a pH-dependent change in calcium affinity alone cannot be responsible for the release of bound protein ligands from the LRP at the pH prevailing in the endosome, which in turn requires one or more other pH dependent effects for regulating protein ligand release. PMID- 11735396 TI - Characterization of a mechanism-based inhibitor of NAD(P)H:quinone oxidoreductase 1 by biochemical, X-ray crystallographic, and mass spectrometric approaches. AB - We report the characterization of 5-methoxy-1,2-dimethyl-3-[(4 nitrophenoxy)methyl]indole-4,7-dione (ES936) as a mechanism-based inhibitor of NQO1. Inactivation of NQO1 by ES936 was time- and concentration-dependent and required the presence of a pyridine nucleotide cofactor consistent with a need for metabolic activation. That ES936 was an efficient inhibitor was demonstrated in these studies by the low partition ratio (1.40 +/- 0.03). The orientation of ES936 in the active site of NQO1 was examined by X-ray crystallography and found to be opposite to that observed for other indolequinones acting as substrates. ES936 was oriented in such a manner that, after enzymatic reduction and loss of a nitrophenol leaving group, a reactive iminium species was located in close proximity to nucleophilic His 162 and Tyr 127 and Tyr 129 residues in the active site. To determine if ES936 was covalently modifying NQO1, ES936-treated protein was analyzed by electrospray ionization liquid chromatography/mass spectrometry (ESI-LC/MS). The control NQO1 protein had a mass of 30864 +/- 6 Da (n = 20, theoretical, 30868.6 Da) which increased by 217 Da after ES936 treatment (31081 +/- 7 Da, n = 20) in the presence of NADH. The shift in mass was consistent with adduction of NQO1 by the reactive iminium derived from ES936 (M + 218 Da). Chymotryptic digestion of the protein followed by LC/MS analysis located a tetrapeptide spanning amino acids 126-129 which was adducted with the reactive iminium species derived from ES936. LC/MS/MS analysis of the peptide fragment confirmed adduction of either Tyr 127 or Tyr 129 residues. This work demonstrates that ES936 is a potent mechanism-based inhibitor of NQO1 and may be a useful tool in defining the role of NQO1 in cellular systems and in vivo. PMID- 11735397 TI - Computational characterization of substrate binding and catalysis in S adenosylhomocysteine hydrolase. AB - S-Adenosylhomocysteine (AdoHcy) hydrolase catalyzes the reversible hydrolysis of AdoHcy to adenosine (Ado) and homocysteine (Hcy), playing an essential role in modulating the cellular Hcy levels and regulating activities of a host of methyltransferases in eukaryotic cells. This enzyme exists in an open conformation (active site unoccupied) and a closed conformation (active site occupied with substrate or inhibitor) [Turner, M. A., Yang, X., Yin, D., Kuczera, K., Borchardt, R. T., and Howell, P. L. (2000) Cell Biochem. Biophys. 33, 101 125]. To investigate the binding of natural substrates during catalysis, the computational docking program AutoDock (with confirming calculations using CHARMM) was used to predict the binding modes of various substrates or inhibitors with the closed and open forms of AdoHcy hydrolase. The results have revealed that the interaction between a substrate and the open form of the enzyme is nonspecific, whereas the binding of the substrate in the closed form is highly specific with the adenine moiety of a substrate as the main recognition factor. Residues Thr57, Glu59, Glu156, Gln181, Lys186, Asp190, Met351, and His35 are involved in substrate binding, which is consistent with the crystal structure. His55 in the docked model appears to participate in the elimination of water from Ado through the interaction with the 5'-OH group of Ado. In the same reaction, Asp131 removes a proton from the 4' position of the substrate after the oxidation reduction reaction in the enzyme. To identify the residues that bind the Hcy moiety, AdoHcy was docked to the closed form of AdoHcy hydrolase. The Hcy tail is predicted to interact with His55, Cys79, Asn80, Asp131, Asp134, and Leu344 in a strained conformation, which may lower the reaction barrier and enhance the catalysis rate. PMID- 11735398 TI - Pulsed-alkylation mass spectrometry for the study of protein folding and dynamics: development and application to the study of a folding/unfolding intermediate of bacterial luciferase. AB - A new method employing the classical techniques of chemical modification of proteins and the new technology of mass spectrometry, known as pulsed-alkylation mass spectrometry (PA/MS), has been developed to probe the dynamic structure of folding intermediates and folded complexes of proteins under a variety of conditions. This method is fast and simple, and the results are easily interpreted. PA/MS may provide an alternative to H/D exchange monitored either by NMR or by electrospray ionization mass spectrometry for some experiments; for others, it may provide access to questions not readily answered by available methods. The objective of PA/MS is to determine simultaneously the location and the extent of labeling of functional groups in a protein by measuring the reactivity of cysteines with N-ethylmaleimide, within the context of the conformation of the protein under specific conditions. The method can also be applied to chemical modification of other amino acid residues employing any of a vast array of reagents, depending upon the specifics of the protein under investigation. The enormous range of reactivity of the thiol groups of the cysteinyl residues in proteins and the change in reactivity upon denaturation or conformational rearrangement afford a large signal change that can be correlated with changes in accessibility of the thiol group. The information obtained from the correlation of observed thiol reactivity with the local environment of each cysteinyl residue in the structure of the folded protein can be supplemented by results obtained from fluorescence, circular dichroism, or other methods, to develop an understanding of the structure and dynamics of altered conformational states. With bacterial luciferase as a model system, we have applied PA/MS to investigate the structural differences between the native heterodimeric enzyme and a folding intermediate that is well-populated in 2 M urea. The thiol residues at positions 307, 324, and 325 of the alpha subunit were much more reactive with N-ethylmaleimide in the presence of 2 M urea than in the native enzyme, suggesting that the C-terminal region of the alpha subunit was less tightly packed in the folding intermediate. The apparent unfolding of the C-terminal region of the alpha subunit of the alphabeta structure in 2 M urea appears to mimic the unfolding of the C-terminal domain of the free alpha subunit, also in 2 M urea, described by Noland, B. W., Dangott, L. J., and Baldwin, T. O. (1999) Biochemistry 38, 16136-16145. The approach described here should be applicable to a wide array of problems that have in common the need to determine the locations of conformational changes in proteins. Application of PA/MS to the investigation of the relative thermodynamic stability of the coordination complexes of zinc within each of the six zinc-finger domains of MRE-binding transcription factor-1 (Zn(6) MTF-zf) in its free and DNA-bound forms is presented in the companion paper in this issue [Apuy, J. L., Chen, X., Russell, D. H., Baldwin, T. O., and Giedroc, D. P. (2001) Biochemistry 40, 15164-15175]. PMID- 11735399 TI - Ratiometric pulsed alkylation/mass spectrometry of the cysteine pairs in individual zinc fingers of MRE-binding transcription factor-1 (MTF-1) as a probe of zinc chelate stability. AB - Metal-response element (MRE)-binding transcription factor-1 (MTF-1) is a zinc regulated transcriptional activator of metallothionein (MT) genes in mammalian cells. The MRE-binding domain of MTF-1 (MTF-zf) has six canonical Cys(2)-His(2) zinc finger domains that are distinguished on the basis of their apparent affinities for zinc and their specific roles in MRE-binding. In this paper, pulsed alkylation of the zinc-liganding cysteine thiolate pairs with the sulfhydryl-specific alkylating reagent d(5)-N-ethylmaleimide (d(5)-NEM) is used as a residue-specific probe of the relative stabilities of the individual zinc finger coordination complexes in Zn(6) MTF-zf. A chase with excess H(5)-N ethylmaleimide (H(5)-NEM) to fully derivatize MTF-zf concomitant with complete proteolysis, followed by MALDI-TOF mass spectrometry allows quantitation of the mole fraction of d(5),d(5)-, d(5),H(5)-, and H(5),H(5)-NEM derivatized peptides corresponding to each individual zinc finger domain as a function of d(5)-NEM pulse time. This experiment establishes the hierarchy of cysteine thiolate reactivity in MTF-zf as F5 > F6 >> F1 > F2 approximately F3 approximately F4. The apparent second-order rate of reaction of F1 thiolates is comparable to that determined for the DNA binding domain of Sp1, Zn(3) Sp1-zf, under identical solution conditions. The reactivities of all Cys residues in MTF-zf are significantly reduced when bound to an MREd-containing oligonucleotide. An identical experiment carried out with Zn(5) MTF-zf26, an MTF-zf domain lacking the N-terminal F1 zinc finger, reveals that MTF-zf26 binds to the MREd very weakly, and is characterized by strongly increased reactivity of nonadjacent F4 thiolates. These findings are discussed in the context of existing models for metalloregulation by MTF-1. PMID- 11735400 TI - Translesional synthesis past acetylaminofluorene-derived DNA adducts catalyzed by human DNA polymerase kappa and Escherichia coli DNA polymerase IV. AB - Human DNA polymerase kappa (pol kappa) has a sequence significantly homologous with that of Escherichia coli DNA polymerase IV (pol IV). We used a truncated form of human pol kappa (pol kappaDeltaC) and full-length pol IV to explore the miscoding properties of these enzymes. Oligodeoxynucleotides, modified site specifically with N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (dG-AAF) and N (deoxyguanosin-8-yl)-2-aminofluorene (dG-AF), were used as DNA templates in primer extension reactions that included all four dNTPs. Reactions catalyzed by pol kappaDeltaC were partially blocked one base prior to dG-AAF or dG-AF, and also opposite both lesions. At higher enzyme concentrations, a significant fraction of primer was extended. Analysis of the fully extended reaction product revealed incorporation of dTMP opposite dG-AAF, accompanied by much smaller amounts of dCMP, dAMP, and dGMP and some one- and two-base deletions. The product terminating 3' to the adduct site contained AMP misincorporated opposite dC. On templates containing dG-AF, dAMP, dTMP, and dCMP were incorporated opposite the lesion in approximately equal amounts, together with some one-base and two-base deletions. Steady-state kinetics analysis confirmed the results obtained from primer extension reactions catalyzed by pol kappa. In contract, primer extension reactions catalyzed by pol IV were blocked effectively by dG-AAF and dG-AF. At high concentrations of pol IV, full-length products were formed containing primarily one- or two-base deletions with dCMP, the correct base, incorporated opposite dG-AF. The miscoding properties of pol kappa observed in this study are consistent with mutational spectra observed when plasmid vectors containing dG AAF or dG-AF are introduced into simian kidney cells [Shibutani, S., et al. (2001) Biochemistry 40, 3717-3722], supporting a model in which pol kappa plays a role in translesion synthesis past acetylaminofluorene-derived lesions in mammalian cells. PMID- 11735401 TI - Electrophoretic characterization of ribosomal subunits and proteins in apoptosis: specific downregulation of S11 in staurosporine-treated human breast carcinoma cells. AB - Stimulation of death receptors (Fas on human T-cell leukemia Jurkat cells and tumor necrosis factor receptor-1 on human monoblastic leukemia U937 cells) triggers the specific degradation of 28S ribosomal RNA, and this process may contribute to cell death through the inhibition of protein synthesis. We have developed an analytical method using a polyacrylamide-agarose composite gel to evaluate ribosomal subunits in apoptotic cells (human breast carcinoma MCF-7 cells treated with staurosporine and human 293T cells irradiated with ultraviolet light were used in addition to the two apoptosis systems described above). No alterations were detected by this method, suggesting that apoptosis, including the process of ribosomal RNA degradation, does not cause fragmentation or extensive conformational changes in the ribosome. We also examined the status of 21 different ribosomal proteins in apoptotic cells by immunoblotting with polyclonal antibodies. S11 was specifically downregulated in apoptotic MCF-7 cells and in other apoptotic breast carcinoma cells. Previous studies have shown that S11 is heterogeneously expressed in cancer cells. Taken together, it appears that particular intracellular environments regulate the expression of S11 protein. However, the mechanism by which this process is modulated is as yet unknown. Furthermore, we have demonstrated that our composite gel electrophoresis system can efficiently detect ubiquitination of ribosomal subunits. PMID- 11735402 TI - Inhibition of the Bloom's and Werner's syndrome helicases by G-quadruplex interacting ligands. AB - G-Quadruplex DNAs are folded, non-Watson-Crick structures that can form within guanine-rich DNA sequences such as telomeric repeats. Previous studies have identified a series of trisubstituted acridine derivatives that are potent and selective ligands for G-quadruplex DNA. These ligands have been shown previously to inhibit the activity of telomerase, the specialized reverse transcriptase that regulates telomere length. The RecQ family of DNA helicases, which includes the Bloom's (BLM) and Werner's (WRN) syndrome gene products, are apparently unique among cellular helicases in their ability to efficiently disrupt G-quadruplex DNA. This property may be relevant to telomere maintenance, since it is known that the sole budding yeast RecQ helicase, Sgs1p, is required for a telomerase independent telomere lengthening pathway reminiscent of the "ALT" pathway in human cells. Here, we show that trisubstituted acridine ligands are potent inhibitors of the helicase activity of the BLM and WRN proteins on both G quadruplex and B-form DNA substrates. Inhibition of helicase activity is associated with both a reduction in the level of binding of the helicase to G quadruplex DNA and a reduction in the degree to which the G-quadruplex DNA can support DNA-dependent ATPase activity. We discuss these results in the context of the possible utility of trisubstituted acridines as antitumor agents for the disruption of both telomerase-dependent and telomerase-independent telomere maintenance. PMID- 11735403 TI - Evidence of powerful substrate electric fields in DNA photolyase: implications for thymidine dimer repair. AB - DNA photolyase is a flavoprotein that repairs cyclobutylpyrimidine dimers by ultrafast photoinduced electron transfer. One unusual feature of this enzyme is the configuration of the FAD cofactor, where the isoalloxazine and adenine rings are nearly in vdW contact. We have measured the steady-state and transient absorption spectra and excited-state decay kinetics of oxidized (FAD-containing, folate-depleted) Escherichia coli DNA photolyase with and without dinucleotide and polynucleotide single-stranded thymidine dimer substrates. The steady-state absorption spectrum for the enzyme-polynucleotide substrate complex showed a blue shift, as seen previously by Jorns et al. (1). No shift was observed for the dinucleotide substrate, suggesting that there are significant differences in the binding geometry of dinucleotide versus polynucleotide dimer lesions. Evidence was obtained from transient absorption experiments for a long-lived charge transfer complex involving the isoalloxazine of the FAD cofactor. No evidence of excited-state quenching was measurable upon binding either substrate. To explain these data, we hypothesize the existence of a large substrate electric field in the cavity containing the FAD cofactor. A calculation of the magnitude and direction of this dipolar electric field is consistent with electrochromic band shifts for both S(0) --> S(1) and S(0) --> S(2) transitions. These observations suggest that the substrate dipolar electric field may be a critical component in its electron-transfer-mediated repair by photolyase and that the unique relative orientation of the isoalloxazine and adenine rings may have resulted from the consequences of the dipolar substrate field. PMID- 11735404 TI - Bypass replication in vitro of UV-induced photoproducts blocking leading or lagging strand synthesis. AB - In vitro replication assays were used to determine the capacity of HeLa extracts to replicate past one of the two major photoproducts produced by ultraviolet radiation at adjacent thymines in duplex DNA, namely, the cis,syn cyclobutane dimer ([c,s]TT) and the 6-4 pyrimidine-pyrimidone adduct ([6-4]TT). The site specific photoproduct was placed on the template either to the leading strand or to the lagging strand of nascent DNA with respect to the first fork encountering the lesion during bidirectional replication of closed circular duplex molecules carrying the SV40 origin. Replication products from time-course reactions were fractionated by gel electrophoresis in the presence of ethidium bromide. Recognition and quantification of true translesion synthesis products, i.e., newly synthesized closed circular molecules carrying the photoproduct, were aided by specific substrate modifications (a T:T mismatch in a unique PstI site nearby the photoproduct) and improved assay conditions (internal standard to control for completion of PstI digestion). Extracts from HeLa cells, which express DNA polymerase eta, were competent to replicate past the [c,s]TT on either strand. The efficiency of bypass replication of the [c,s]TT on the template to the leading or the lagging strand was 71% and 67%, respectively. The same extracts demonstrated very low efficiency of translesion synthesis (at most 8-10%) of the [6-4]TT on either template position. Replication-competent cell-free extracts from other human cells were also deficient in the bypass of the [6-4]TT in vitro. PMID- 11735405 TI - Role of SRP RNA in the GTPase cycles of Ffh and FtsY. AB - The bacterial homologues of the signal recognition particle (SRP) and its receptor, the Ffh*4.5S RNA ribonucleoprotein complex and the FtsY protein, respectively, form a unique complex in which both Ffh and FtsY act as GTPase activating proteins for one another, resulting in the mutual stimulation of GTP hydrolysis by both proteins. Previous work showed that 4.5S RNA enhances the GTPase activity in the presence of both Ffh and FtsY, but it was not clear how this was accomplished. In this work, kinetic and thermodynamic analyses of the GTPase reactions of Ffh and FtsY have provided insights into the role of 4.5S RNA in the GTPase cycles of Ffh and FtsY. We found that 4.5S RNA accelerates the association between Ffh and FtsY 400-fold in their GTP-bound form, analogous to its 200-fold catalytic effect on Ffh*FtsY association previously observed with the GppNHp-bound form [Peluso, P., et al. (2000) Science 288, 1640-1643]. Further, Ffh-FtsY association is rate-limiting for the observed GTPase reaction with subsaturating Ffh and FtsY, thereby accounting for the apparent stimulatory effect of 4.5S RNA on the GTPase activity observed previously. An additional step, GTP hydrolysis from the Ffh*FtsY complex, is also moderately facilitated by 4.5S RNA. These results suggest that 4.5S RNA modulates the conformation of the Ffh*FtsY complex and may, in turn, regulate its GTPase activity during the SRP functional cycle. PMID- 11735406 TI - Apoflavodoxin folding mechanism: an alpha/beta protein with an essentially off pathway intermediate. AB - The folding reaction of Anabaena apoflavodoxin has been studied by stopped-flow kinetics and site-directed mutagenesis. Although the urea unfolding equilibrium is two-state, a transient intermediate accumulates during the folding reaction. The intermediate is monomeric, and it is not related to proline isomerization. Unlike many cases where the presence of an intermediate has been detected either by a burst phase or by the curvature, at low urea concentration, of the otherwise only observable kinetic phase, two kinetic phases are observed in apoflavodoxin folding whose total amplitude equals the amplitude of unfolding. To determine the role of the intermediate in the folding reaction, the apoflavodoxin kinetic data have been fitted to all conceivable three-species kinetic models (either linear or triangular). Using a stepwise fitting procedure, we find that the off-pathway mechanism explains most of the kinetic data (not a slow unfolding phase), the on pathway mechanism being rejected. By using global analysis, good overall agreement between data and fit is found when a triangular mechanism is considered. The fitted values of the microscopic constants indicate that most of the unfolded molecules refold from the denatured state. Apoflavodoxin thus folds via a triangular, but essentially off-pathway, mechanism. We calculate that the retardation of the folding caused by the off-pathway intermediate is not large. Some unusual properties of the intermediate are discussed. PMID- 11735407 TI - Three-dimensional solution structure and stability of thioredoxin m from spinach. AB - Proton NMR spectral resonances of thioredoxin m from spinach have been assigned, and its solution structure has been determined on the basis of 1156 nuclear Overhauser effect- (NOE-) derived distance constraints by using restrained molecular dynamics calculations. The average pairwise root-mean-square deviation (RMSD) for the 25 best NMR structures for the backbone was 1.0 +/- 0.1, when the structurally well-defined residues were considered. The N- and C-terminal segments (1-13 and 118-119) and residues 41-49, comprising the active site, are highly disordered. At the time of concluding this work, a crystal structure of this protein was reported, in which thioredoxin m was found to crystallize as noncovalent dimers. Although the solution and crystal structures are very similar, no evidence was found about the existence of dimers in solution, thus confirming that dimerization is not needed for the regulatory activity of thioredoxin m. The spinach thioredoxin m does not unfold by heat in the range 25 85 degrees C, as revealed by thermal circular dichroic (CD) measurements. However, its unfolding free energy (9.1 +/- 0.8 kcal mol(-1), at pH 5.3 and 25 degrees C) could be determined by extrapolating the free energy values obtained at different concentrations of guanidinium chloride (GdmCl). The folding unfolding process is two-state as indicated by the coincidence of the CD denaturation curves obtained at far and near UV. The H/D exchange behavior of backbone amide protons was analyzed. The slowest-exchanging protons, requiring a global-unfolding mechanism in order to exchange, are those from beta2, beta3, and beta4, the central strands of the beta-sheet, which constitute the main element of the core of the protein. The free energies obtained from exchange measurements of protons belonging to the alpha-helices are lower than those derived from GdmCl denaturation studies, indicating that those protons exchange by local-unfolding mechanisms. PMID- 11735408 TI - Slow folding of three-fingered toxins is associated with the accumulation of native disulfide-bonded intermediates. AB - Snake neurotoxins are short all-beta proteins that display a complex organization of the disulfide bonds: two bonds connect consecutive cysteine residues (C43-C54, C55-C60), and two bonds intersect when bridging (C3-C24, C17-C41) to form a particular structure classified as "disulfide beta-cross". We investigated the oxidative folding of a neurotoxin variant, named alpha62, to define the chemical nature of the three-disulfide intermediates that accumulate during the process in order to describe in detail its folding pathway. These folding intermediates were separated by reverse-phase HPLC, and their disulfide bonds were identified using a combination of tryptic hydrolysis, manual Edman degradation, and mass spectrometry. Two dominant intermediates containing three native disulfide bonds were identified, lacking the C43-C54 and C17-C41 pairing and therefore named des [43-54] and des-[17-41], respectively. Both species were individually allowed to reoxidize under folding conditions, showing that des-[17-41] was a fast-forming nonproductive intermediate that had to interconvert into the des-[43-54] isomer before forming the native protein. Conversely, the des-[43-54] intermediate appeared to be the immediate precursor of the oxidized neurotoxin. A kinetic model for the folding of neurotoxin alpha62 which fits with the observed time course accumulation of des-[17-41] and des-[43-54] is proposed. The effect of turn 2, located between residues 17 and 24, on the overall kinetics is discussed in view of this model. PMID- 11735409 TI - pH-jump-induced folding and unfolding studies of barstar: evidence for multiple folding and unfolding pathways. AB - Equilibrium and kinetic characterization of the high pH-induced unfolding transition of the small protein barstar have been carried out in the pH range 7 12. A mutant form of barstar, containing a single tryptophan, Trp 53, completely buried in the core of the native protein, has been used. It is shown that the protein undergoes reversible unfolding above pH 10. The pH 12 form (the D form) appears to be as unfolded as the form unfolded by 6 M guanidine hydrochloride (GdnHCl) at pH 7 (the U form): both forms have similar fluorescence and far-UV circular dichroism (CD) signals and have similar sizes, as determined by dynamic light scattering and size-exclusion chromatography. No residual structure is detected in the D form: addition of GdnHCl does not alter its fluorescence and far-UV CD properties. The fluorescence signal of Trp 53 has been used to monitor folding and unfolding kinetics. The kinetics of folding of the D form in the pH range 7-11 are complex and are described by four exponential processes, as are the kinetics of unfolding of the native state (N state) in the pH range 10.5-12. Each kinetic phase of folding decreases in rate with increase in pH from 7 to 10.85, and each kinetic phase of unfolding decreases in rate with decrease in pH from 12 to 10.85. At pH 10.85, the folding and unfolding rates for any particular kinetic phase are identical and minimal. The two slowest phases of folding and unfolding have identical kinetics whether measured by Trp 53 fluorescence or by mean residue ellipticity at 222 nm. Direct determination of the increase in the N state with time of folding at pH 7 and of the D form with time of unfolding at pH 12, by means of double-jump assays, show that between 85 and 95% of protein molecules fold or unfold via fast pathways between the two forms. The remaining 5 15% of protein molecules appear to fold or unfold via slower pathways, on which at least two intermediates accumulate. The mechanism of folding from the high pH denatured D form is remarkably similar to the mechanism of folding from the urea or GdnHCl-denatured U form. PMID- 11735410 TI - Packing is a key selection factor in the evolution of protein hydrophobic cores. AB - The energy derived from optimized van der Waals interactions in closely packed, folded proteins has been proposed to be of similar energetic magnitude to hydrophobicity in stabilizing the native state. If packing is this energetically important, it should influence the evolution of protein core sequences. To test this hypothesis, the occurrence of various amino acid side chains in the major hydrophobic core of staphylococcal nuclease and 42 homologous proteins was determined. Most such positions in this protein family are usually isoleucine, leucine, or valine. Previously we have constructed and measured the stabilities of 12 single, 44 double, 64 triple, and 32 quadruple mutants, representing all possible permutations of these three side chains at two overlapping sets of four positions in the core of staphylococcal nuclease. The stabilities and interaction energies of those mutants with various combinations of the most common, or consensus, sequence were compared to the stabilities of all other mutants. Mutants which had the consensus side-chain combinations were not necessarily the most stable, but usually were found to have the best interaction energies. In other words, these proteins were far more stable than would be predicted from simply summing the observed energetic effects of the component single mutations, apparently reflecting particularly favorable packing interactions that are possible for the most common side chains. An additional 12 mutants which tested possible alternate explanations of the results were constructed. The stabilities and interaction energies of these mutants also support the conclusion that packing is a crucial determinant guiding the sequence evolution of protein cores. PMID- 11735411 TI - Production of phosphatidylinositol 3,4,5-trisphosphate and phosphatidic acid in platelet rafts: evidence for a critical role of cholesterol-enriched domains in human platelet activation. AB - Glycosphingolipid- and cholesterol-enriched membrane microdomains, called rafts, can be isolated from several mammalian cells, including platelets. These microdomains appear to play a critical role in signal transduction in several hematopoietic cells, but their function in blood platelets remains unknown. Herein, we first characterized the lipid composition, including the fatty acid composition of phospholipids, of human platelet rafts. Then their role in platelet activation process was investigated. Interestingly, thrombin stimulation led to morphological changes of rafts correlating with the production of lipid second messengers in these microdomains. Indeed, we could demonstrate for the first time that a large part of the stimulation-dependent production of phosphatidic acid and phosphoinositide 3-kinase products was concentrated in rafts. Moreover, cholesterol depletion with methyl-beta-cyclodextrin disrupted platelet rafts, dramatically decreased the agonist-dependent production of these lipid signaling molecules, and impaired platelet secretion and aggregation. Cholesterol repletion restored the physiological platelet responses. Altogether our data indicate that rafts are highly dynamic platelet membrane structures involved in critical signaling mechanisms linked to the production of lipid second messengers. The demonstration of phosphatidylinositol 3,4,5-trisphosphate production in rafts may have general implications for the understanding of the role of this key second messenger found ubiquitously in higher eucaryotic cells. PMID- 11735412 TI - Mechanism of peroxynitrite interaction with ferric hemoglobin and identification of nitrated tyrosine residues. CO(2) inhibits heme-catalyzed scavenging and isomerization. AB - Hemoproteins are one of the major targets of peroxynitrite in vivo. It has been proposed that the bimolecular heme/peroxynitrite interaction results in both peroxynitrite inactivation (scavenging) and catalysis of tyrosine nitration. In this study, we used spectroscopic techniques to analyze the reaction of peroxynitrite with human methemoglobin (metHb). Although conventional differential spectroscopy did not reveal heme changes, our results suggest that, in the absence of bicarbonate, the heme in metHb reacts bimolecularly with peroxynitrite but is quickly back-reduced by the reaction products. This hypothesis is based on two indirect observations. First, metHb prevents the peroxynitrite-mediated nitration of a target dipeptide, Ala-Tyr, and second, it promotes the isomerization of peroxynitrite to nitrate. Both the scavenging and the isomerization activities of metHb were heme-dependent and inhibited by CO(2). Ferrous cytochrome c was an efficient scavenger of peroxynitrite, but in the ferric form did not show either scavenging or isomerization activities. We found no evidence of an increase in Ala-Tyr nitration with these hemoproteins. Peroxynitrite-treated metHb induced the formation of a long-lived radical assigned to tyrosine by spin-trapping studies. This radical, however, did not allow us to predict an interaction of peroxynitrite with heme. Hb was nitrated by peroxynitrite/CO(2) mainly in tyrosines beta 130, alpha 42, and alpha 140 and, to a lesser extent, alpha 24. The nitration of alpha chain tyrosines more exposed to the solvent (alpha 140 and alpha 24) was higher in CO-Hb and metHb, while nitration of alpha 42, the tyrosine nearest to the heme, was higher in oxyHb. We deduce that the heme/peroxynitrite interaction, which is inhibited in CO-Hb and metHb, affects alpha tyrosine nitration in two opposite ways, i.e., by protecting exposed residues and by promoting nitration of the residue nearest to the heme. Conversely, nitration of beta Tyr 130 was comparable in oxyHb, metHb, and CO-Hb, suggesting a mechanism involving only nitrating species formed during peroxynitrite decay. PMID- 11735413 TI - Identification of homologous biologically active sites on the N-terminal domain of laminin alpha chains. AB - Laminin, a multifunctional glycoprotein of the basement membrane, consists of three different subunits, alpha, beta, and gamma chains. To date, five different alpha chains have been identified. N-terminal domain VI in the alpha1 chain has various biological activities. Here we screened biologically active sequences on domain VI of the laminin alpha2, alpha3, and alpha5 chains using a large number of overlapping peptides. HT-1080 human fibrosarcoma cell attachment to the peptides was evaluated using peptide-coated plastic plates and peptide-conjugated Sepharose beads. We identified four cell adhesive sequences from laminin alpha2 chain domain VI, two sequences from the alpha3 chain, and two sequences from the laminin alpha5 chain. Sequences homologous to A13 (RQVFQVAYIIIKA, alpha1 chain 121-133) on all the alpha chains (FQIAYVIVKA, alpha2 chain 130-139; GQLFHVAYILIKF, alpha3 chain 96-108; FHVAYVLIKA, alpha5 chain 74-83) showed strong cell attachment activity. A5-16 (LENGEIVVSLVNGR, alpha5 chain 147-160) showed the strongest cell attachment activity in the plate assay, and the homologous peptide in the alpha3 chain promoted similar strong cell attachment activity. A5-16 and its homologous peptide in the alpha2 chain promoted moderate cell attachment, while the homologous peptide to A5-16 in the alpha1 chain did not show activity. A2-7 (SPSIKNGVEYHYV, alpha2 chain 108-120) showed cell attachment activity only in the plate assay, but homologous sequences in the alpha1, alpha3, and alpha5 chains did not promote activity. A2-7 promoted endothelial cell sprouting from aortic rings in vitro and melanoma colonization to murine lungs in vivo. However, none of the homologous peptides of A2-7 promoted experimental pulmonary metastasis by B16-BL6 melanoma cells. These results indicate that there are chain specific active sites in domain VI of the laminin alpha chains, some of which contain conserved activities. PMID- 11735414 TI - Selective activation of the 20 S proteasome (multicatalytic proteinase complex) by histone h3. AB - Two distinct activities cleaving bonds after hydrophobic amino acids have been identified in the bovine pituitary 20 S proteasome. One, expressed by the X subunit, that cleaves bonds after aromatic and branched chain amino acids was designated as chymotrypsin-like (ChT-L).(1) The second, expressed by the Y subunit, that cleaves bonds after acidic amino acids was designated as peptidylglutamyl-peptide hydrolyzing (PGPH) but also cleaves bonds after branched chain amino acids. Low micromolar concentrations of the arginine-rich histone H3 (H3) are shown to induce changes in the specificity of the proteasome by selectively activating cleavages after branched chain and acidic amino acids while inhibiting cleavage of peptidyl-arylamide bonds in synthetic substrates. H3 activates 15-fold cleavage after leucine but not phenylalanine residues in model synthetic substrates. The activation is associated with a decrease in K(m) and an increase in V(max), suggesting positive allosteric activation. H3 activates more than 60-fold degradation of the oxidized B-chain of insulin, by cleaving mainly bonds after acidic and branched chain amino acids, and accelerates the degradation of casein and lysozyme, the latter in the presence of dithiothreitol. The degradation of lysozyme in the presence of H3 generates fragments that differ from those in its absence, indicating H3-induced specificity changes. H3 inhibits cleavage of the Trp3-Ser4 and Tyr5-Gly6 bonds in gonadotropin releasing hormone, bonds cleaved by the ChT-L activity in the absence of H3. The results suggest H3 selective activation of the Y subunit and specificity changes that could potentially affect proteasomal function in the nuclear compartment. PMID- 11735415 TI - Role of the disulfide cleavage induced molten globule state of type a botulinum neurotoxin in its endopeptidase activity. AB - Botulinum neurotoxins are produced by anaerobic Clostridium botulinum in an inactive form. The endopeptidase activity of type A botulinum neurotoxin (BoNT/A) is triggered by reduction of its disulfide bond between its heavy chain and light chain. By using circular dichroism spectroscopy, we show that, upon reduction of BoNT/A and under physiological temperature (37 degrees C), the BoNT/A loses most of its native tertiary structure, while retaining most of its secondary structure. This type of structure is characterized as a molten globule type conformation, which was further confirmed for BoNT/A by the characteristic binding of 1-anilinonaphthalene-8-sulfonic acid. Under nonreducing conditions where the interchain disulfide bond is intact, the enzymatically inactive BoNT/A did not show a molten globule type of structure. A temperature profile of the structure and enzyme activity of BoNT/A revealed that, under reducing conditions, there was a strong correlation in the existence of the molten globule structure and optimum endopeptidase activity at about 37 degrees C. PMID- 11735416 TI - Calbindin D(9k): a protein optimized for calcium binding at neutral pH. AB - The binding of calcium ions by EF-hand proteins depends strongly on the electrostatic interactions between Ca(2+) ions and negatively charged residues of these proteins. We have investigated the pH dependence of the binding of Ca(2+) ions by calbindin D(9k). This protein offers a unique possibility for interpretation of such data since the pK(a) values of all ionizable groups are known. The binding is independent of pH between 7 and 9, where maximum calcium affinity is observed. An abrupt decrease in the binding affinity is observed at pH values below 7. This decrease is due to protonation of acidic groups, leading to modification of protein charges. The pH dependence of the product of the two macroscopic Ca(2+)-binding constants can be formally described by the involvement of two acidic groups with pK(a) = 6.6. Monte Carlo calculations show that the reduction of Ca(2+) binding is strictly determined by variable electrostatic interactions due to pH-dependent changes not only in the binding sites, but also of the overall charge of the protein. PMID- 11735417 TI - Characterization of hK4 (prostase), a prostate-specific serine protease: activation of the precursor of prostate specific antigen (pro-PSA) and single chain urokinase-type plasminogen activator and degradation of prostatic acid phosphatase. AB - hK4 (prostase, KLK4), a recently cloned prostate-specific serine protease and a member of the tissue kallikrein family, is a zymogen composed of 228 amino acid residues including an amino-terminal propiece, Ser-Cys-Ser-Gln-. A chimeric form of hK4 (ch-hK4) was constructed in which the propiece of hK4 was replaced by that of prostate-specific antigen (PSA) to create an activation site susceptible to trypsin-type proteases. ch-hK4 was expressed in Escherichia coli, isolated from inclusion bodies, refolded, and purified with an overall yield of 25%. The zymogen was readily self-activated during the refolding process to generate an active form (21 kDa) of hK4 (rhK4). rhK4 cleaved the chromogenic substrates Val Leu-Arg-pNA (S-2266), Pro-Phe-Arg-pNA (S-2302), Ile-Glu-Gly-Arg-pNA (S-2222), and Val-Leu-Lys-pNA (S-2251), indicating that rhK4 has a trypsin-type substrate specificity. The rhK4 was inhibited by aprotinin (6 kDa), forming an equimolar 27 kDa complex. rhK4 readily activated both the precursor of PSA (pro-PSA) and single chain urokinase-type plasminogen activator (scuPA, pro-uPA). rhK4 also completely degraded prostatic acid phosphatase but failed to cleave serum albumin, another protein purified from human seminal plasma. These results indicate that hK4 may have a role in the physiologic processing of seminal plasma proteins such as pro-PSA, as well as in the pathogenesis of prostate cancer through its activation of pro-uPA. PMID- 11735418 TI - Peroxidase- and nitrite-dependent metabolism of the anthracycline anticancer agents daunorubicin and doxorubicin. AB - Oxidation of the anticancer anthracyclines doxorubicin (DXR) and daunorubicin (DNR) by lactoperoxidase(LPO)/H(2)O(2) and horseradish peroxidase(HRP)/H(2)O(2) systems in the presence and absence of nitrite (NO(2)(-)) has been investigated using spectrophotometric and EPR techniques. We report that LPO/H(2)O(2)/NO(2)(-) causes rapid and irreversible loss of anthracyclines' absorption bands, suggesting oxidative degradation of their chromophores. Both the initial rate and the extent of oxidation are dependent on both NO(2)(-) concentration and pH. The initial rate decreases when the pH is changed from 7 to 5, and the reaction virtually stops at pH 5. Oxidation of a model hydroquinone compound, 2,5-di-tert butylhydroquinone, by LPO/H(2)O(2) is also dependent on NO(2)(-); however, in contrast to DNR and DXR, this oxidation is most efficient at pH 5, indicating that LPO/H(2)O(2)/NO(2)(-) is capable of efficiently oxidizing simple hydroquinones even in the neutral form. Oxidation of anthracyclines by HRP/H(2)O(2)/NO(2)(-) is substantially less efficient relative to that by LPO/H(2)O(2)/NO(2)(-) at either pH 5 or pH 7, most likely due to the lower rate of NO(2)(-) metabolism by HRP/H(2)O(2). EPR measurements show that interaction of anthracyclines and 2,5-di-tert-butylhydroquinone with LPO/H(2)O(2)/NO(2)(-) generates the corresponding semiquinone radicals presumably via one-electron oxidation of their hydroquinone moieties. The possible role of the (*)NO(2) radical, a putative LPO metabolite of NO(2)(-), in oxidation of these compounds is discussed. Because in vivo the anthracyclines may co-localize with peroxidases, H(2)O(2), and NO(2)(-) in tissues, their oxidation via the proposed mechanism is likely. These observations reveal a novel, peroxidase- and nitrite dependent mechanism for the oxidative transformation of the anticancer anthracyclines, which may be pertinent to their biological activities in vivo. PMID- 11735419 TI - Tryptophan and tyrosine radicals in ribonucleotide reductase: a comparative high field EPR study at 94 GHz. AB - Tryptophan radicals, which are generated in the reconstitution reaction of mutants Y122F and Y177W of subunit R2 apoprotein of E. coli and mouse ribonucleotide reductase (RNR), respectively, with Fe(2+) and oxygen, are investigated by high-field EPR at 94 GHz and compared with the tyrosine radicals occurring in the respective wild-type proteins. For the first time, accurate g values are obtained for protein-associated neutral tryptophan free radicals, which show only a small anisotropy. The apparent hyperfine patterns observed in frozen solutions are very similar for tryptophan and tyrosine radicals in mouse subunit R2 at conventional X-band EPR. The radicals can, however, be discriminated by their different g-tensors using high-field EPR. Tryptophan radicals were postulated as reaction intermediates in the proposed radical transfer pathway of RNR. Furthermore, the data obtained here for the electronic structure of protein-associated tryptophan neutral free radicals are important for identification and understanding of the functional important tryptophan radicals which occur in other enzymes, e.g., DNA photolyase and cytochrome c peroxidase, where they are magnetically coupled to other radicals or to a metal center. PMID- 11735420 TI - Characterization of the interaction between androgen receptor and a new transcriptional inhibitor, SHP. AB - SHP (short heterodimer partner) is an orphan nuclear receptor, first described for its interaction with nuclear receptors. This study explores a new way of inhibiting the androgen-signaling pathway. We demonstrated that SHP inhibited up to 97% of AR-induced activity. Characterization of AR/SHP interaction provided evidence of a clear ligand dependency. We also showed that the LXXI/LL motifs previously found on SHP mediated the interaction with the AR ligand-binding domain (AR-LBD), the motif responsible for the interaction being slightly different from that found with ER. The AR N-terminal domain (AR-NTD), in contrast to that of other nuclear receptors, accounts for most of the entire receptor transactivation potential. SHP also interacted with AR-NTD, thus stabilizing the interaction with AR. We demonstrated that SHP inhibited both AR-LBD and NTD dependent transactivation, which evidenced for the first time a protein capable of inhibiting a steroid receptor amino-terminal-dependent transactivation. We further characterized the SHP mechanism of action by showing that SHP reversed AR coactivator-mediated activation. Conversely, FHL2 and TIF2 counteracted SHP mediated inhibition of AR. SHP evidences a new way of inhibiting AR activity by competing with AR coactivators. This new type of inhibitor could dictate the activity of nuclear receptors, depending on the equilibrium between activators and inhibitors. PMID- 11735421 TI - Probing DNA single strands for single-base bulges with neocarzinostatin chromophore. AB - Neocarzinostatin chromophore (NCS-Chrom) induces strong cleavage at a single site (C3) in the single-stranded and 5' (32)P-end-labeled 13-mer GCCAGATTTGAGC in a reaction dependent on a thiol. By contrast, in the duplex form of the same 13 mer, strand cleavage occurs only at the T and A residues, and C3 is not cleaved. To determine the minimal structural requirement(s) for C3 cleavage in the single stranded oligomer, several deletions and mutations were made in the 13-mer. A 10 mer (GCCAGAGAGC) derived from the 13-mer by deletion of the three T residues was also cleaved exclusively at C3 by NCS-Chrom, generating fragments having 5' phosphate ends. That the cleavage at C3 is initiated by abstraction of its 5' hydrogen is confirmed in experiments using 3' (32)P-end-labeled 10-mer. The competent 13-mer and 10-mer were assigned hairpin structures with a stem loop and a single bulged out A base, placing C3 across from and 3' to the bulge. Removal of the bulged A base from the 13-mer and the 10-mer resulted in complete loss of cutting activity, proving that it is the essential determinant in competent substrates. Studies of thiol post-activated NCS-Chrom binding to the DNA oligomers show that the drug binds to the bulge-containing 13-mer (K(d) = 0.78 microM) and the 10-mer (K(d) = 1.11 microM), much more strongly than to the 12 mer (K(d) = 20 microM) and the 9-mer (K(d) = 41 microM), lacking the single-base bulge. A mutually induced-fit between NCS-Chrom and the oligomer resulting in optimal stabilization of the drug-DNA complex is proposed to account for the site specific cleavage at C3. These studies establish the usefulness of NCS-Chrom as a probe for single-base bulges in DNA. PMID- 11735422 TI - A reaction-induced FT-IR study of cyanobacterial photosystem I. AB - In oxygenic photosynthesis, photosystem I (PSI) conducts light-driven electron transfer from plastocyanin to ferredoxin. The reactions are initiated when the primary chlorophyll donor, P(700), is photooxidized. P(700) is a chlorophyll dimer ligated by the core subunits psaA and psaB. A difference Fourier transform infrared spectrum, associated with P(700)(+)-minus-P(700), can be acquired using PSI from the cyanobacterium Synechocystis sp. PCC 6803. This spectrum reflects contributions from oxidation-sensitive modes of chlorophyll, as well as from oxidation-induced structural changes in amino acid residues and the peptide backbone. Oxidation-induced structural changes may play a role in the facilitation and control of electron-transfer reactions involving the primary donor. In this paper, we report that photooxidation of P(700) in cyanobacterial PSI perturbs a cysteine residue. At 264 and 80 K, a downshift of a SH stretching vibration from 2560 to 2551 cm(-1) is observed. Such a downshift is consistent with an increase in hydrogen bonding, with a change in C-S-H conformation, or with an electric field effect. Deuterium exchange experiments were also performed. While the perturbed cysteine is in a protein region that is resistant to exchange, other (2)H-sensitive vibrational chl and amino acid bands were observed. From the (2)H exchange experiments, we conclude that photooxidation of P(700) perturbs internal or bound water molecules in PSI and that the P(700)(+) minus-P(700) spectrum is (2)H exchange-sensitive. The results are consistent with structural complexity in the PSI primary donor, as previously suggested [Kim, S., and Barry, B. A. (2000) J. Am. Chem. Soc. 122, 4980-4981]. Possible explanations, including a partial enolization of P(700)(+), are discussed. PMID- 11735423 TI - Carboxyl residues in the iron-sulfur protein are involved in the proton pumping activity of P. denitrificans bc(1) complex. AB - A study is presented on chemical modification of the three subunit Paracoccus denitrificans bc(1) complex. N-(Ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline (EEDQ) treatment caused a loss of the proton pumping activity of liposome reconstituted bc(1) complex. A similar effect, which is referred to as the decoupling effect, resulted upon reaction of N,N'-dicyclohexylcarbodiimide (DCCD) with the complex. Direct measurement of the binding of EEDQ to the complex subunits, performed in the presence of the fluorescent hydrophobic nucleophile 4' [(aminoacetamido)methyl]fluorescein (AMF), showed that the iron-sulfur protein (ISP) and cytochrome c(1) were labeled by EEDQ, whereas cytochrome b was not. Tryptic digestion and sequencing analysis of the fluorescent fragment of the ISP revealed this to consist of a segment with six acidic residues, among which the highly conserved aspartate 160 is present. Analogous experiments on DCCD binding showed that all the three subunits of the complex were labeled. However, DCCD concentration dependence of carboxyl residue modification in the individual subunits and of proton pumping activity showed that the decrease of the H(+)/e(-) ratio correlated only with the modification of the ISP. Tryptic digestion of labeled ISP and sequencing analysis of the fluorescent fragment gave results superimposable upon those obtained with EEDQ. Chymotryptic digestion and sequencing analysis of the single fluorescent fragment of cytochrome b showed that this fragment contained glutamate 174 and aspartate 187. We conclude that, in the P. denitrificans bc(1) complex, carboxyl residues in cytochrome b do not appear to be critically involved in the proton pump mechanism of the complex. PMID- 11735424 TI - Electrostatic interactions between charged amino acid residues and the bacteriochlorophyll dimer in reaction centers from Rhodobacter sphaeroides. AB - The extent of electrostatic contributions from the protein environment was assessed by the introduction of ionizable residues near the bacteriochlorophyll dimer in reaction centers from Rhodobacter sphaeroides. Two mutations at symmetry related sites, M199 Asn to Asp and L170 Asn to Asp, resulted in a 48 and 44 mV lowering of the midpoint potential, respectively, compared to the wild type at pH 8, while a 75 mV decrease in the midpoint potential was observed for the mutation L168 His to Glu. The decrease relative to wild type was found to be approximately additive, up to 147 mV, for various combinations of the mutations. As the pH was lowered from 9.5 to 6.0, the relative decrease in the midpoint potential became smaller for each of these three mutations. Titration of the pH dependence of the change in midpoint potential of the M199 Asn to Asp mutant compared to wild type yielded a pK(a) value of 7.9 and a change in midpoint potential from low to high pH of 59 mV. The major effect of the mutation on the midpoint potential of the dimer is interpreted as stemming from a negative charge on the residue. An average dielectric constant of approximately 20 was estimated for the local protein environment, consistent with a relatively hydrophobic environment for residue M199. The rate of charge recombination between the primary quinone acceptor and the bacteriochlorophyll dimer decreased in the M199 Asn to Asp mutant at high pH, reflecting the decrease in midpoint potential. PMID- 11735425 TI - Dominant thermodynamic role of the third independent receptor binding site in the receptor-associated protein RAP. AB - The 39 kDa receptor-associated protein (RAP) is a three-domain escort protein in the secretory pathway for several members of the low-density lipoprotein receptor (LDLR) family of endocytic receptors, including the LDLR-related protein (LRP). The minimal functional unit of LRP required for efficient binding to RAP is composed of complement-type repeat (CR)-domain pairs, located in clusters on the extracellular part of LRP. Here we investigate the binding of full-length RAP and isolated RAP domains 1-3 to an ubiquitin-fused CR-domain pair consisting of the fifth and sixth CR domains of LRP (U-CR56). As shown by isothermal titration calorimetric analysis of simple RAP domains as well as adjoined RAP domains, all three RAP domains bind to this CR-domain pair in a noncooperative way. The binding of U-CR56 to RAP domains 1 and 2 is (at room temperature) enthalpically driven with an entropy penalty (K(D) = 2.77 x 10(-6) M and 1.85 x 10(-5) M, respectively), whereas RAP domain 3 binds with a substantially lower enthalpy, but is favored due to a positive entropic contribution (K(D) = 1.71 x 10(-7) M). The heat capacity change for complex formation between RAP domain 1 and the CR domain pair is -1.65 kJ K(-1) mol(-1). There is an indication of a conformational change in RAP domain 3 upon binding in the surface plasmon resonance analysis of the interaction. The different mechanisms of binding to RAP domains 1 and 3 are further substantiated by the different effects on binding of mutations of the Asp and Trp residues in the LRP CR5 or CR6 domains, which are important for the recognition of several ligands. PMID- 11735426 TI - G(s) signaling is intact after disruption of lipid rafts. AB - Membrane microdomains enriched in cholesterol and sphingolipids modulate a number of signal transduction pathways and provide a residence for heterotrimeric G proteins, their receptors, and their effectors. We investigated whether signaling through G(s) was dependent on these membrane domains, characterized by their resistance to detergents, by depleting cells of cholesterol and sphingolipids. For cholesterol depletion, rat salivary epithelial A5 cells were cultured under low-cholesterol conditions, and then treated with the cholesterol chelator methyl beta-cyclodextrin. For sphingolipid depletion, LY-B cells, a mutant CHO cell line that is unable to synthesize sphingolipids, were incubated under low-sphingolipid conditions. Depletion of cholesterol or sphingolipid led to a loss or decrease, respectively, in the amount of Galpha(s) from the detergent-resistant membranes without any change in the cellular or membrane-bound amounts of Galpha(s). The cAMP accumulation in response to a receptor agonist was intact and the level slightly increased in cells depleted of cholesterol or sphingolipids compared to that in control cells. These results indicate that localization of Galpha(s) to detergent-resistant membranes was not required for G(s) signaling. Analysis of the role of lipid rafts on the kinetics of protein associations in the membrane suggests that compartmentalization in lipid rafts may be more effective in inhibiting protein interactions and, depending on the pathway, ultimately inhibit or promote signaling. PMID- 11735427 TI - Photoaffinity labeling of human sex hormone-binding globulin using 17alpha alkylamine derivatives of 3beta-androstanediol substituted with azidonitrophenylamido, azidonitrophenylamino, or trifluoroazidonitrophenylamino chromophores. Localization of Trp-84 in the vicinity of the steroid-binding site. AB - Purified human SHBG was photoaffinity labeled with 17alpha-aminomethyl (M), 17alpha-aminoethyl (E), and 17alpha-aminopropyl (P) derivatives of [3alpha-(3)H] 5alpha-androstane-3beta,17beta-diol coupled to 5-azido-2-nitrobenzoylamido (ANB), 4-azido-2-nitrophenylamino (ANP), and 5-azido-2-nitro-3,4,6-trifluorophenylamino (ANTFP) chromophores. Successful labeling was achieved in all cases except for the two photoreagents with the shortest side chains, namely, ANP-M and ANTFP-M derivatives. Edman sequencing and mass spectrometry of immunopurified photolabeled tryptic fragments revealed that radioactivity was present either on the sequence of residues 73-94, uniquely at the level of Trp-84 (stable covalent labeling), or on one of the two overlapping sequences of residues 126-134 and 126 135, at the level of Pro-130 (labile labeling) and Lys-134 (either stable or partially labile labeling), respectively. The same Trp-84 was photolabeled with the three ANB derivatives of increasing lengths, and by the ANP-P photoreagent. This residue was the exclusive target for the shortest [(3)H]ANB-M photoreagent but was a minor site for the longest [(3)H]ANB-P photoreagent, essentially recovered at the level of Pro-130. The [(3)H]ANB-E photoreagent of intermediate size also labeled exclusively Trp-84, except in some experiments in which photolabeling was recovered predominantly at the level of Pro-130. The [(3)H]ANP P photoreagent with an overall length similar to that of the ANB-P photoreagent labeled simultaneously Trp-84 (minor site) and Lys-134. The other [(3)H]ANP-E, [(3)H]ANTFP-E, and [(3)H]ANTFP-P derivatives labeled in all cases Lys-134. These findings indicate that the conserved Trp-84 and the two Pro-130 and Lys-134 residues are all located in the vicinity of the D ring of steroid ligands and remain freely accessible from the C17alpha position, thus providing biochemical data delineating the corresponding region of the steroid-binding site. PMID- 11735428 TI - Functional implications of the unstructured loop in the (beta/alpha)(8) barrel structure of the bacterial luciferase alpha subunit. AB - Bacterial luciferase catalyzes the conversion of FMNH(2), a long-chain aliphatic aldehyde, and molecular oxygen to FMN, the corresponding carboxylic acid, and H(2)O with the emission of light. The light-emitting species is an enzyme-bound excited state flavin. The enzyme is a heterodimer (alphabeta) of homologous subunits each with an (beta/alpha)(8) barrel structure. A portion of the loop in the alpha subunit that connects beta strand 7 to alpha helix 7 is disordered in the crystal structure. To test the hypothesis that this loop closes over the active site during catalysis and protects the active site from bulk solvent, a mutant was constructed in which the 29 residues that are disordered in the 2.4 A crystal structure were deleted. Deletion of this loop results in a heterodimer with a subunit equilibrium dissociation constant of 1.32 +/- 1.25 microM, whereas the wild-type heterodimer shows no measurable subunit dissociation. This mutant retains its ability to bind substrate flavin and aldehyde with wild-type affinity and can carry out the chemistry of the bioluminescence reaction with nearly wild type efficiency. However, the bioluminescent quantum yield of the reaction is reduced nearly 2 orders of magnitude from that of the wild-type enzyme. PMID- 11735429 TI - Oxidation-reduction and activation properties of chloroplast fructose 1,6 bisphosphatase with mutated regulatory site. AB - The concentration of Mg(2+) required for optimal activity of chloroplast fructose 1,6-bisphosphatase (FBPase) decreases when a disulfide, located on a flexible loop containing three conserved cysteines, is reduced by the ferredoxin/thioredoxin system. Mutation of either one of two regulatory cysteines in this loop (Cys155 and Cys174 in spinach FBPase) produces an enzyme with a S(0.5) for Mg(2+) (0.6 mM) identical to that observed for the reduced WT enzyme and significantly lower than the S(0.5) of 12.2 mM of oxidized WT enzyme. E(m) for the regulatory disulfide in WT spinach FBPase is -305 mV at pH 7.0, with an E(m) vs pH dependence of -59 mV/pH unit, from pH 5.5 to 8.5. Aerobic storage of the C174S mutant produces a nonphysiological Cys155/Cys179 disulfide, rendering the enzyme partially dependent on activation by thioredoxin. Circular dichroism spectra and thiol titrations provide supporting evidence for the formation of nonphysiological disulfide bonds. Mutation of Cys179, the third conserved cysteine, produces FBPase that behaves very much like WT enzyme but which is more rapidly activated by thioredoxin f, perhaps because the E(m) of the regulatory disulfide in the mutant has been increased to -290 mV (isopotential with thioredoxin f). Structural changes in the regulatory loop lower S(0.5) for Mg(2+) to 3.2 mM for the oxidized C179S mutant. These results indicate that opening the regulatory disulfide bridge, either through reduction or mutation, produces structural changes that greatly decrease S(0.5) for Mg(2+) and that only two of the conserved cysteines play a physiological role in regulation of FBPase. PMID- 11735431 TI - Effects of cloned gene dosage on the response of recombinant CHO cells to hyperosmotic pressure in regard to cell growth and antibody production. AB - The effect of cloned gene dosage on growth and product formation under hyperosmotic conditions has been studied using recombinant Chinese hamster ovary (rCHO) cell lines producing chimeric antibody. Batch cultures of four rCHO cell lines carrying different numbers of antibody gene copies were carried out using the hyperosmolar medium. Depending on cloned gene dosage, hyperosmotic pressure decreased specific growth rate (mu) and increased specific antibody productivity (q(Ab)) to a different degree. The cell line with lower cloned gene dosage displayed more significant enhancement in q(Ab) and less reduction in mu at hyperosmolalities. However, the cell line with higher cloned gene dosage still yielded higher maximum antibody concentration at hyperosmolality up to 469 mOsm/kg. Northern blot analysis showed a positive relationship between immunoglobulin mRNA level per cell and q(Ab), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Cell cycle analysis showed that hyperosmotic pressure induced G(1)-phase arrest, suggesting that the increase of cell population in G(1)-phase may contribute in part to enhanced q(Ab) at hyperosmolality. Taken together, although the cell line with lower cloned gene dosage displayed more significant enhancement in q(Ab) at hyperosmolality, the factor that determined the maximum antibody concentration in hyperosmotic rCHO cell cultures was almost exclusively the gene dosage. PMID- 11735430 TI - Strategies for enhancing monoclonal antibody accumulation in plant cell and organ cultures. AB - Various strategies aimed at improving IgG(1) antibody accumulation in transgenic tobacco cell and organ cultures were tested. The form of tissue had a significant effect on antibody levels; shooty teratomas were less productive than hairy roots or suspended cells. Although there were several disadvantages associated with hairy roots compared with suspensions, such as slower growth, slower antibody production, and formation of a greater number of antibody fragments, the roots exhibited superior long-term culture stability. Antibody accumulation in hairy root cultures was improved by increasing the dissolved oxygen tension to 150% air saturation, indicating the need for effective oxygen transfer in root reactors used for antibody production. Preventing N-linked glycosylation using tunicamycin or inhibition of subsequent glycan processing by castanospermine reduced antibody accumulation in the biomass and/or medium in cell suspensions. Loss of antibody from the cultures after its secretion and release into the medium was identified as a major problem. This effect was minimized by inhibiting protein transport in the secretory pathway using Brefeldin A, resulting in antibody accumulation levels up to 2.7 times those in untreated cells. Strategies for protecting secreted antibody, such as addition of poly(vinylpyrrolidone) and periodic harvesting from the medium using hydroxyapatite resin, also increased antibody titers. The mechanisms responsible for the disappearance of antibody from plant culture media were not clearly identified; degradation by proteases and conformational modification of the antibody, such as formation of aggregates, provided an explanation for some but not all the phenomena observed. This work demonstrates that the manipulation and control of culture conditions and metabolic processes in plant tissue cultures can be used to improve the production of foreign proteins. However, loss of secreted antibody from plant culture medium is a significant problem that may limit the feasibility of using product recovery from the medium to reduce downstream processing costs relative to agricultural systems. PMID- 11735432 TI - Simultaneous analysis of spatio-temporal gene expression for cephamycin biosynthesis in Streptomyces clavuligerus. AB - Linkage between structural and regulatory genes implies that a direct correlation should exist between the spatio-temporal distribution of their expression. Green fluorescent protein (GFP) and cyan fluorescent protein (CFP) were used as reporters to analyze simultaneously expression of lysine-epsilon-aminotransferase (LAT) and its corresponding genetic regulator, CcaR. The isogenic strain containing lat::gfp and ccaR::cfp in the chromosome produced cephamycin C at levels similar to wild type Streptomyces clavuligerus. Confocal laser scanning microscopy revealed that expression of both LAT and CcaR in liquid culture was temporally dynamic and spatially heterogeneous in S. clavuligerus mycelia. During the early culture stage only a part of the mycelia began to express LAT and CcaR at low levels. As the culture aged, expression levels and the population of mycelia expressing LAT and CcaR increased and were followed late in the growth cycle by a reduction of the mycelia population expressing LAT and CcaR. The approach provides a precise simultaneous temporal-spatial expression profile and corroborates the regulatory linkage between ccaR and lat in S. clavuligerus. PMID- 11735433 TI - Influence of sulfobetaines on the stability of the Citrobacterdiversus ULA-27 beta-lactamase. AB - The activity and stability of beta-lactamase from Citrobacter diversus ULA-27 have been investigated in the presence of different ionic and zwitterionic surfactants. All the sulfobetaine surfactants tested allow the enzyme to retain its full activity, but the best stabilizing effect is greatly dependent on their structure. Very little variations on the monomer headgroup can significantly reduce enzyme deactivation or speed up the loss of activity with respect to buffer alone. The whole hydrophobic/hydrophilic balance on the headgroup seems to have a determining role in preserving beta-lactamase activity and structure. The presence of zwitterionic surfactants stabilizes the protein conformation toward denaturation by urea and low-temperature inactivation. Similar experiments were performed in the presence of other two zwitterionic surfactants, an amine oxide, dimethylmyristylamine oxide (DMMAO) and a carboxybetaine, cetyldimethylammonium methanecarboxylate (CB1-16). The former stabilizes the enzyme even better than the sulfobetaines, the latter quickly deactivates it. Therefore, the factors responsible for beta-lactamase stabilization are dependent not only on the zwitterionic nature of the surfactant headgroup but also specific interactions between the surfactant and the protein may be important. PMID- 11735434 TI - Optimization of fermentation reaction conditions for preparation of PATE enzyme by Erwinia carotovora IFO3830 by TFCCRD method. AB - The fermentation conditions for preparation of polygalacturonic acid transeliminase (PATE) enzyme by Erwinia carotovora IFO3830 were optimized for seed ratio, vibration rate, and temperature by the TFCCRD method. The results indicated that the optimum fermentation conditions for E. carotovora IFO 3830 were that seed ratio, vibration rate, and temperature were 5% v/v, 113 min(-1), and 29 degrees C, respectively. PMID- 11735435 TI - Biphasic inactivation kinetics of Escherichia coli in liquid whole egg by high hydrostatic pressure treatments. AB - Kinetic studies on the isothermal high hydrostatic pressure (HHP) inactivation of Escherichia coli in liquid whole egg (LWE) were performed at 5 and 25 degrees C in the pressure range of 250-400 MPa. The characteristic tailing inactivation curves were described by a first-order biphasic model. As compared to a previous rheological study, it is suggested that the phase change of LWE during pressure treatment affects the inactivation rate of E. coli. Within the processing criteria where the rheological properties of LWE were still comparable to those of fresh LWE, HHP treatments at 5 degrees C induced more E. coli inactivations than those at 25 degrees C. From the results of approximately 3 log reductions of E. coli and over 5 log reductions of Pseudomonas and Paenibacillus, HHP treatment of LWE at 5 degrees C is regarded to be as effective as conventional thermal pasteurization. However, no post-process contamination and the consistency of temperature during preparation, HHP treatment, and storage provide clear processing advantages. PMID- 11735436 TI - Interaction of 3-hydroxybenzoate with 3-hydroxybenzoate-6-hydroxylase. AB - The gradual quenching of the emission fluorescence of 3-HBA in the visible region upon titration with 3-HBA-6-hydroxylase and distinct changes in the near-UV circular dichroic spectrum of the enzyme in the presence of substrate suggest the formation of a stable enzyme-substrate complex. The binding of aromatic substrate 3-hydroxybenzoate to 3-hydroxybenzoate-6-hydroxylase occurs without gross changes in the backbone structure of the enzyme. The binding strength of the ES complex is partially reduced upon chemical modification of arginine, histidine, or tryptophan residues of enzyme, probably implicating their concerted action in the binding of substrate to enzyme. Partial inactivation of enzyme and diminished stability of the ES complex in response to treatment with 1 M urea could be ascribed to localized effects of the denaturant. PMID- 11735437 TI - Analysis of CHO cells metabolic redistribution in a glutamate-based defined medium in continuous culture. AB - The effect of glutamine replacement by glutamate and the balance between glutamate and glucose metabolism on the redistribution of t-PA-producing recombinant CHO cells metabolism is studied in a series of glucose shift down and shift up experiments in continuous culture. These experiments reveal the existence of multiple steady states, and experimental data are used to perform metabolic flux analysis to gain a better insight into cellular metabolism and its redistribution. Regulation of glucose feed rate promotes a higher efficiency of glucose and nitrogen source utilization, with lower production of metabolic byproducts, but this reduces t-PA specific production rate. This reduction under glucose limitation can be attributed to the fact that the cells are forced to efficiently utilize the carbon and energy source for growth, impairing the production of dispensable metabolites. It is, therefore, the combination of growth rate and carbon and energy source availability that determines the level of t-PA production in continuous culture. PMID- 11735438 TI - Effect of oscillating dissolved oxygen tension on the production of alginate by Azotobacter vinelandii. AB - The effect of oscillating dissolved oxygen tension (DOT) on the metabolism of an exopolysaccharide-producing bacteria (Azotobacter vinelandii) was investigated, particularly on the mean molecular weight (MMW) of the alginate produced. Sinusoidal DOT oscillations were attained by manipulating the oxygen and nitrogen partial pressures at the inlet of a 1.0 L working volume bioreactor. Periods of 1200, 2400, and 4000 s and average amplitudes between 1.0% and 2.2% DOT, with an oscillation axis fixed at 3% DOT, were tested. A culture carried out at constant 3% DOT was used as comparison. The average wave amplitude had an important effect on the maximum mean molecular weight (MMW(max)) of the alginate produced. The higher the amplitude, the lower the MMW(max). As the average wave amplitudes decreased from 2.2% to 1.0%, the MMW(max) increased from 64 to 240 KDa, respectively. Furthermore, at 3% constant DOT (0.0% of amplitude), a MMW(max) of 350 KDa was obtained. No important effect of the oscillating DOT on kinetics of biomass growth, alginate production, and sucrose consumption was observed, compared with constant DOT. The findings of this study point out that accurate DOT control is crucial if a particular molecular weight species of alginate needs to be produced, particularly in large fermentors, where bacteria are exposed to an oscillatory environment as a result of DOT gradients caused by the high viscosity of the broth and insufficient mixing. PMID- 11735439 TI - Do enzymatic hydrolyzability and Simons' stain reflect the changes in the accessibility of lignocellulosic substrates to cellulase enzymes? AB - In an attempt to elucidate the impact of substrate accessibility to cellulases on the susceptibility of lignocellulosic substrates to enzymatic hydrolysis, a hydrogen peroxide treated, Douglas fir kraft pulp was dried using several methods with varying levels of intensity. Oven-drying at 50 and 100 degrees C, air drying, and freeze-drying methods were employed to remove the interfibrillar water from the pulp samples. Subsequently, the never-dried and variably dried pulps were hydrolyzed using a commercial cellulase preparation supplemented with additional beta-glucosidase. Drying reduced the susceptibility of the substrates to enzymatic hydrolysis, which can be attributed to the hornifying effect that drying has on fibers. This effect was more pronounced for the fibers that were oven-dried at 100 degrees C (23% reduction) and 50 degrees C (15% reduction), and there was a good correlation between the Simons's stain results and the enzymatic digestibility of the dried pulps. These observations indicated that drying significantly reduced the population of larger pores and that the partial closure of larger pores created a large number of smaller pores that were not accessible to the displacement dye molecules (orange dye). The inaccessibility of the cellulose to the enzymes, due to the collapse or closure of the large pores, appears to be the primary reason for the lower susceptibility of the dried pulps to enzymatic hydrolysis. PMID- 11735440 TI - Metabolic flux variation of Saccharomyces cerevisiae cultivated in a multistage continuous stirred tank reactor fermentation environment. AB - The technique of metabolic flux analysis was implemented to elucidate the flux balancing of Saccharomyces cerevisiae cultivated in a multistage continuous stirred tank reactor fermentation environment. The results showed that the majority of the substrate (97.70 +/- 0.49%) was funneled into the glycolytic pathway, while the remainder was subdivided between the pentose phosphate pathway and pathways for polysaccharide synthesis. At the pyruvate node, 87.30 +/- 1.38% of the flux was channeled through the reaction governed by pyruvate decarboxylase. Fluxes through the pyruvate dehydrogenase bypass were maintained at a constant level (82.65 +/- 1.47%) irrespective of the configuration of the fermentation setup. Activity through the TCA "cycle" was replenished by the reaction catalyzed by pyruvate carboxylase and by the transport of cytosolic oxaloacetate across the mitochondrial membrane. The CO(2) evolution rate varied as fermentation progressed; however, the yield coefficient of CO(2) remained at a constant value. Although a constant yield of ethanol (0.42 g of ethanol/g of glucose) was obtained, operations of the TCA cycle were gradually switched from partially reductive to partially oxidative pathways from the first fermenter to the fourth fermenter. PMID- 11735441 TI - Rice straw as a support for immobilization of microbial lipase. AB - Candida rugosa lipase was covalently immobilized on rice straw activated with glutaraldehyde using poly(ethylene glycol) (PEG) as the stabilizing agent. The effects of PEG molecular weight and enzyme loading were studied according to a full 2(2) factorial design. Higher immobilization yields (>70%) were attained when the lipase loading was 95 units/mg of dry support, independent of PEG molecular weight. All derivatives showed high hydrolytic and synthetic activities. This work provides preliminary results on the use of agricultural residues as a support matrix for immobilizing lipase and on the application of the resulting derivatives to butyl butyrate synthesis as a study model. PMID- 11735442 TI - Packed bed column fermenter and kinetic modeling for upgrading the nutritional quality of coffee husk in solid-state fermentation. AB - Studies were carried out to evaluate solid-state fermentation (SSF) for the upgradation of the nutritional quality of coffee husk by degrading the caffeine and tannins present in it. SSF was carried out by Aspergillus niger LPBx in a glass column fermenter using factorial design experiments and surface response methodology to optimize bioprocess parameters such as the substrate pH and moisture content and aeration rate. The first factorial design showed that the moisture content of the substrate and aeration rate were significant factors for the degradation of toxic compounds, which was confirmed by the second factorial design too. The kinetic study showed that the degradation of toxic compounds was related to the development of the mold and its respiration and also to the consumption of the reducing sugars present in coffee husk. From the values obtained experimentally for the oxygen uptake rate and CO(2) evolved, the system determined a biomass yield (Y(x/o)) of 3.811 (g of biomass).(g of consumed O(2))( 1) and a maintenance coefficient (m) of 0.0031 (g of consumed O(2)).(g biomass of biomass)(-1).h(-1). The best results on the degradation of caffeine (90%) and tannins (57%) were achieved when SSF was carried out with a 30 mL.min(-1) aeration rate using coffee husk having a 55% initial moisture content. The inoculation rate did not affect the metabolization of the toxic compounds by the fungal culture. After SSF, the protein content of the husk was increased to 10.6%, which was more than double that of the unfermented husk (5.2%). PMID- 11735443 TI - Effect of inoculum composition and low KCl supplementation on the biological and rheological stability of an immobilized-cell system for mixed mesophilic lactic starter production. AB - Two strains of Lactococcus lactis subsp. lactis (L. lactis KB and KBP) and one of L. lactis subsp. lactis biovar. diacetylactis (L. diacetylactis MD) were immobilized separately in kappa-carrageenan-locust bean gum gel beads. Continuous fermentations were carried out in supplemented whey permeate in a 1-L pH controlled stirred tank reactor inoculated with a 30% (v/v) bead inoculum and a bead ratio of 55:30:15 for KB, KBP, and MD, respectively. The process demonstrated a high productivity and microbial stability during the 7-week continuous culture. Compared with previous experiments carried out with an inoculum bead ratio of 33:33:33 for KB, KBP, and MD beads, respectively, the modification of the inoculum bead ratio had apparently little effect on free and immobilized, total and specific populations. A dominant behavior of L. diacetylactis MD over the other strains of the mixed culture was observed both with free-cell populations in the effluent and with immobilized-cell populations. Additional experiments were carried out with other strain combinations for continuous inoculation-prefermentation of milk. The data also confirmed the dominance of L. diacetylactis during long-term continuous immobilized-cell fermentations. This dominance may be tentatively explained by the local competition involved in the development of the bead cross-contamination and in citrate utilization by L. diacetylactis strains. The gel beads demonstrated a high rheological stability during the 7-week continuous fermentation even at low KCl supplementation of the broth medium (25 mM KCl). PMID- 11735444 TI - Fluidized bed design parameters affecting novel lactic Acid downstream processing. AB - Lactic acid purification was directly done from fermentation utilizing a fluidized bed column refilled with a strong anionic exchange resin. The purpose of this work was to study the influence of two important design parameters, bed diameter (D) and bed-height (H), in the lactic acid binding and elution capacity of the matrix. By changing the settled bed height from 2.5 to 5 cm for each diameter of column analyzed it was possible to obtain an 50% increase in the binding capacity of the resin in all experiments. This fact was attributed to a higher contact time between the culture broth and the anionic resin produced by the increase of back mixing and lactic acid residence time. PMID- 11735445 TI - Feasibility study of a compact process for biological treatment of highly soluble VOCs polluted gaseous effluent. AB - Volatile organic compounds (VOCs), representing a wide range of products mainly generated by industrial activity, are involved in air pollution. This study deals with a new biological treatment process of gaseous effluent combining a gas/liquid contactor called an "aero-ejector" and a membrane bioreactor. Combining these two innovative technologies enables a high elimination efficiency to be reached. We first focus on transfer phenomena characterization in a pilot installation on a laboratory scale, using a gaseous effluent polluted with a low ethanol concentration (7.1 x 10(-3) kg.m(-3)). These experiments demonstrated the good transfer performances since 90% of the ethanol was absorbed in the liquid phase in one step. After this physical characterization, the biological aspect of the system was studied using the yeast Candida utilis as microorganism. During the experiment, no ethanol was measured in the fermentation broth nor in the outlet gas, confirming the efficiency of ethanol elimination by C. utilis. The experimental procedure emerging from the present study strongly validates the suitability of this process for ethanol removal from air. PMID- 11735446 TI - Enhancing protein capacity of rigid macroporous polymeric adsorbent. AB - A macroporous poly(glycidyl methacrylate-triallyl isocyanurate-divinylbenzene) resin was synthesized and modified with diethylamine to yield an anion-exchange resin suitable for protein adsorption. Efforts were made to enhance protein ion exchange capacity of the resin by investigating the copolymer composition. Different synthesis recipes were attempted, and the resultant resins were characterized by measuring the specific surface area and the adsorption ability using bovine serum albumin (BSA) as a model protein. The intraparticle pore size distribution measured by mercury porosimetry showed that the pores in the range of 40-120 nm took 75% of the total pore volume, indicating that the ion exchanger was favorable for protein adsorption. BSA capacity obtained with an appropriate recipe was as high as 78.6 mg/g wet resin or 50 mg/mL packed volume, which was higher than the capacities of some commercially available ion exchangers. Moreover, by using a pore diffusion model, the effective pore diffusivity of BSA was found to be 5.5 x 10(-12) m(2)/s, similar to those in the commercial ion exchangers. PMID- 11735448 TI - Selective precipitation of DNA by spermine during the chemical extraction of insoluble cytoplasmic protein. AB - The direct chemical extraction of recombinant L1 protein (the major capsid protein of human papillomavirus type 16) from the cytoplasm of E. coli HMS174(DE3) has recently been demonstrated at high cell density (to OD(600) = 160) without the use of reducing agent (1). Coextraction of DNA at high concentration prevents direct coupling to postextraction recovery operations including expanded bed adsorption. In this study, spermine is used to selectively precipitate DNA during chemical extraction. Highly efficient and selective DNA precipitation was achieved. An approximate 10-fold increase in the specific spermine concentration (mg of spermine/mg of DNA) was required to precipitate DNA when 8 M urea was added to the extraction buffer. EDTA (3 mM), required for effective chemical extraction, does not significantly inhibit DNA precipitation. Precipitation selectivity was demonstrated in a bovine serum albumin spiking test, with almost complete recovery of the spiked protein. During studies on the direct extraction of L1 protein from cells at OD(600) = 80, high DNA removal efficiency (>85%) and negligible L1 protein coprecipitation were achieved. This selective precipitation technique simply requires the addition of spermine to the chemical extraction buffer and therefore does not increase technique complexity. This modification enhances the method's general applicability and enables direct coupling to downstream recovery units following chemical extraction at high cell and product concentrations. PMID- 11735447 TI - Physicochemical properties and structural changes in vegetative tissues as affected by a direct current electrical field. AB - Cylindrical pieces of potato, sweet potato, kohlrabi, radish, and pear were interposed between a pair of electrodes, and a direct current was applied. A special custom-made apparatus enabled the use of differently shaped electrodes. The electrical field was applied for 1 min at 40 V/cm and caused a reduction in specimen weight by a minimal value of 2.7% of initial weight in sweet potato to a maximum 38.4% in pear. The affected area of the tissue resembled the shape of the electrode. Pores were produced in the tissue (from the anode side), possibly promoting slow release of minerals and other cell components from the contracted specimens. From the cathode side, cell "sealing" could be observed. Weight loss was dependent on the mechanical properties of the nontreated vegetative tissue specimens. After confirmation that all samples pass through induced electrical shrinkage, further work, executed only on potato, demonstrated that after electrical treatment the samples were less brown (higher L values). In addition, a dependence of weight loss on current intensity, electrode diameter, and surface ratio between the electrode and specimen was shown. The reduction in weight loss could be useful for initial drying of vegetative materials. Indirect proof of a decrease in enzyme activity as a result of electrical field application could be beneficial in replacing traditional methods for browning prevention. PMID- 11735449 TI - Performances of the immunomagnetic separation method for Cryptosporidium in water under various operation conditions. AB - Immunomagnetic separation (IMS) has been specified as a standard method for the measurement of Cryptosporidium in some countries. In this study, the IMS method was evaluated on the basis of the recovery efficiencies of Cryptosporidium oocysts at various IMS operation conditions. The average recovery for different Cryptosporidium concentrations in deionized water was 82.6 +/- 18.2% (n = 52). No significant change in recovery was observed by altering the debris ratio of the water samples. The efficiency was increased by prolonging the reaction time, and by increasing the amount of immunomagnetic beads. The recoveries of oocysts seeded in an Eppendorf with a small reaction volume were similar to those seeded in glass tubes with 10 times the reaction volume. The recovery efficiency of oocysts was reduced significantly when the reaction buffer was replaced by PBS. In conclusion, this method has good reproducibility and high recovery. PMID- 11735450 TI - Purification of recombinant human B-domain-deleted factor VIII using anti-factor VIII monoclonal antibody selected by the surface plasmon resonance biosensor. AB - The surface plasmon resonance (SPR) biosensor measures the real-time kinetics of noncovalent interaction between a receptor and its ligand. Monoclonal antibodies (mAbs) against recombinant factor VIII (rFVIII) were screened from 127 mAb candidates using the SPR biosensor for the purpose of affinity purification of rFVIII. Each mAb showed a different association and dissociation capacity for rFVIII at each buffer condition. One mAb, F8-38, was selected for immunopurification of rFVIII. To characterize the selected mAb F8-38, the immunopurification results on the anti-FVIII mAb F8-38 affinity gel and the anti von Willebrand factor (vWF) mAb affinity gel were studied. Immunopurification by the anti-vWF affinity gel showed a lower binding capacity of rFVIII and resulted in low purification efficiency. On the other hand, immunopurification by the anti FVIII affinity gel exhibited a 3.5-fold binding capacity and a 2-fold purification efficiency compared to those of the anti-vWF affinity gel. The amounts of proteins and DNAs derived from host cells and mouse IgGs derived from the affinity matrix in the affinity eluate were similar to those of the anti-vWF affinity gel. In conclusion, the SPR method of immunopurification is a useful technology in the screening of mAbs aimed at the development of an affinity purification procedure, and the mAb F8-38 was selected using this technology on the basis of the purification procedure of rFVIII. PMID- 11735451 TI - The use of ion-selective electrodes for evaluating residence time distributions in expanded bed adsorption systems. AB - The suitability of ion-selective electrodes (ISE) for the determination of residence time distribution (RTD) in turbid, cell-containing fluids was examined. The electrodes were found to give reproducible signals in biomass-containing feedstock with up to 20% wet weight of solids. The enhanced feedstock compatibility of IES, when compared to other tracer sensing devices, allows the study of expanded bed system hydrodynamics under relevant operating conditions. Within the linear range of the corresponding ISE-tracer pair, both examined ISE (Li(+)- or Br(-)-selective) showed to be insensitive against the range of flow rate and pH normally employed during expanded bed adsorption (EBA) of proteins. Analyzing the RTD obtained after a perfect ion tracer pulse in terms of the PDE model (PDE, axially dispersed plug-flow exchanging mass with stagnant zones) gave a quantitative description of the underlying hydrodynamic situation during EBA processing. These data provided a powerful tool to make predictions on the adsorptive global process performance with a defined feedstock type and composition. The link between the hydrodynamic events during feedstock application and the actual process performance was shown when applying intact yeast cell suspensions at different biomass content (up to 7.5% wet weight) and buffer conductivity (5-12 mS) onto an EBA column filled with the adsorbent Streamline Q XL as fluidized phase. On the basis of our experimental results, a guideline for the successful application of the ISE/RTD method to EBA process design is presented. PMID- 11735452 TI - Proteomic investigation of metabolic shift in mammalian cell culture. AB - Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures. PMID- 11735453 TI - Separation of a breast cancer cell line from human blood using a quadrupole magnetic flow sorter. AB - We have developed a quadrupole magnetic flow sorter (QMS) to facilitate high throughput binary cell separation. Optimized QMS operation requires the adjustment of three flow parameters based on the immunomagnetic characteristics of the target cell sample. To overcome the inefficiency of semiempirical operation/optimization of QMS flow parameters, a theoretical model of the QMS sorting process was developed. Application of this model requires measurement of the magnetophoretic mobility distribution of the cell sample by the cell tracking velocimetry (CTV) technique developed in our laboratory. In this work, the theoretical model was experimentally tested using breast carcinoma cells (HCC1954) overexpressing the HER-2/neu gene, and peripheral blood leukocytes (PBLs). The magnetophoretic mobility distribution of immunomagnetically labeled HCC1954 cells was measured using the CTV technique, and then theoretical predictions of sorting recoveries were calculated. Mean magnetophoretic mobilities of (1-3) x 10(-4) mm(3)/(T A s) were obtained depending on the labeling conditions. Labeled HCC1954 cells were mixed with unlabeled PBLs to form a "spiked" sample to be separated by the QMS. Fractional recoveries of cells for different flow parameters were examined and compared with theoretical predictions. Experimental results showed that the theoretical model accurately predicted fractional recoveries of HCC1954 cells. High-throughput (3.29 x 10(5) cells/s) separations with high recovery (0.89) of HCC1954 cells were achieved. PMID- 11735454 TI - Quantifying viral propagation in vitro: toward a method for characterization of complex phenotypes. AB - For a eukaryotic virus to successfully infect and propagate in cultured cells several events must occur: the virion must identify and bind to its cellular receptor, become internalized, uncoat, synthesize viral proteins, replicate its genome, assemble progeny virions, and exit the host cell. While these events are taking place, intrinsic host defenses activate in order to defeat the virus, e.g., activation of the interferon system, induction of apoptosis, and attempted elicitation of immune responses via chemokine and cytokine production. As a first step in developing an imaging methodology to facilitate direct observation of such complex host/virus dynamics, we have designed an immunofluorescence-based system that extends the traditional plaque assay, permitting simultaneous quantification of the rate of viral spread, as indicated by the presence of a labeled viral protein, and cell death in vitro, as indicated by cell loss. We propose that our propagation and cell death profiles serve as phenotypic read outs, complementing genetic analysis of viral strains. As our virus/host system we used vesicular stomatitis virus (VSV) propagating in hamster kidney epithelial (BHK-21) and murine astrocytoma (DBT) cell lines. Viral propagation and death profiles were strikingly different in these two cell lines, displaying both very different initial titer and cell age effects. The rate of viral spread and cell death tracked reliably in both cell lines. In BHK-21 cells, the rate of viral propagation, as well as maximal spread, was relatively insensitive to initial titer and was roughly linear over several days. In contrast, viral plaque expansion in DBT cells was contained early in the infections with high titers, while low titer infections spread in a manner similar to the BHK-21 cells. The effect of cell age on infection spread was negligible in BHK-21 cells but not in DBTs. Neither of these effects was clearly observed by plaque assay. PMID- 11735455 TI - Shear deactivation of cellulase, exoglucanase, endoglucanase, and beta glucosidase in a mechanically agitated reactor. AB - Shear deactivation of cellulase and its major component enzymes, viz., exoglucanase (exo-1,4-beta-D-glucan-4-cellobiohydrolase), endoglucanase (endo-1,4 beta-D-glucanhydrolase), and 1,4-beta-glucosidase, was carried out by exposing cellulase to shear in a mechanically agitated reactor in the presence as well as in the absence of the substrate cellulose. Cellulase was found to undergo deactivation when subjected to shear, and the extent of deactivation increased with increasing speed of agitation. Among the three major component enzymes of cellulase, exoglucanase showed rapid deactivation and contributed the most to cellulase deactivation. The presence of a substrate did not affect the deactivation of cellulase. PMID- 11735456 TI - Engineering the microstructure and permeability of thin multilayer latex biocatalytic coatings containing E. coli. AB - The microstructure and permeability of rehydrated 20-100 microm thick partially coalesced (vinyl-actetate acrylic copolymer) SF091 latex coatings and a 118 microm thick model trilayer biocatalytic coating consisting of two sealant SF091 layers containing a middle layer of viable E. coli HB101 + latex were studied as delaminated films in a diffusion apparatus with KNO(3) as the diffussant. The permeability of the hydrated coatings is due to diffusive transport through the pore space between the partially coalesced SF091 latex particles. Coating microstructure was visualized by fast freeze cryogenic scanning electron microscopy (cryo-SEM). The effective diffusion coefficient of SF091 latex coatings (diffusive permeability/film thickness) was determined as the ratio of the effective diffusivity of KNO(3) to its diffusivity in water (D(eff)/D). Polymer particle coalescence was arrested by two methods to increase coating permeability. The first used glycerol with coating drying at 4 degrees C, near the glass transition temperature (T(g)). The second method used sucrose or trehalose as a filler to arrest coalescence; the filler was then dissolved away. D(eff)/D was measured as a function of film thickness; content of glycerol, sucrose, and trehalose; drying time; and rehydration time. D(eff)/D varied from 3 x 10(-4) for unmodified SF091 coatings to 6.8 x 10(-2) for coatings containing sucrose. D(eff)/D was reduced by the flattening of latex particles against the surface of the solid substrate, as well as by the presence of the colloid stabilizer hydroxyethylcellulose (HEC). When corrected for the flattened particle layer, D(eff)/D of HEC-free coatings was as high as 0.20, which agreed with the value predicted from analysis of cryo-SEM images of the coat surface. D(eff)/D decreased by one-half in approximately 5 days in rehydrated SF091 coatings, indicating that significant wet coalescence occurs after glycerol, sucrose, or trehalose are leached from the films. D(eff)/D of SF091 latex trilayer coatings containing viable E. coli HB101 cells decreased as cell loading was increased from 2.2 x 10(-2) for 64 g dry cell weight per liter of coat volume to 5 x 10(-3) for 151 g DCW/L of coat volume. The reduction in coating permeability with increasing cell loading is predicted by Maxwell's equation for D(eff)/D in periodic composites. PMID- 11735458 TI - The electronic structure of non-heme iron(III)-hydroperoxo and iron(III)-peroxo model complexes studied by Mossbauer and electron paramagnetic resonance spectroscopies. PMID- 11735457 TI - Analysis of biofilm structure and gene expression using fluorescence dual labeling. AB - The use of biofilms for the degradation of recalcitrant environmental contaminants or for the production of secondary metabolites necessitates understanding and controlling gene expression. In this work, dual labeling with green fluorescent protein (GFP) variants was used to investigate inducible gene expression in a biofilm. Colocalization of GFP emissions was used to determine regions of attached cells and to correlate structure and activity within the biofilm. The labeling strategy reported here is unique in that the two GFP signals were distinguished by differential excitation rather than differential emission. PMID- 11735459 TI - Cu(II) coordination complex involving TTF-py as ligand. PMID- 11735460 TI - Toward the design of double metal cyanides for the copolymerization of CO(2) and epoxides. PMID- 11735461 TI - Supramolecular architecture of a silver(I) coordination polymer supported by a new ligand containing four Tris(pyrazolyl)methane units. PMID- 11735462 TI - 31P NMR probes of chemical dynamics: paramagnetic relaxation enhancement of the (1)H and (31)P NMR resonances of methyl phosphite and methylethyl phosphate anions by selected metal complexes. AB - Methyl phosphite ((CH(3)O)P(H)(O)(2)(-); MeOPH) and methylethyl phosphate ((CH(3)O)P(OCH(2)CH(3))(O)(2)(-); MEP) are two members of a class of anionic ligands whose (31)P T(2) relaxation rates are remarkably sensitive to paramagnetic metal ions. The temperature dependence of the (31)P NMR line broadenings caused by the Mn(H(2)O)(6)(2+) ion and a water-soluble manganese(III) porphyrin (Mn(III)TMPyP(5+)) indicates that the extent of paramagnetic relaxation enhancement is a measure of the rate at which the anionic probes come into physical contact with the paramagnetic center (i.e., enter the inner coordination shell); that is, piDeltanu(par) = k(assn)[M], where Deltanu(par) is the difference between the line widths of the resonance in paramagnetic and diamagnetic solutions, and k(assn) is the second-order rate constant for association of the phosphorus ligand with the metal, M. Comparison of the (31)P T(1) and T(2) relaxation enhancements shows that rapid T(2) relaxation by the metal ion is caused by scalar interaction with the electronic spin. Relaxation of the phosphorus-bound proton of MeOPH ((1)H-P) by Mn(III)TMPyP(5+) displayed intermediate exchange kinetics over much of the observable temperature range. The field strength dependence of (1)H-P T(2) enhancement and the independence of the (31)P T(2) support these assertions. As in the case of the (31)P T(2), the (1)H-P T(2) relaxation enhancement results from scalar interaction with the electronic spin. The scalar coupling interpretation of the NMR data is supported by a pulsed EPR study of the interactions of Mn(H(2)O)(6)(2+) with the P-deuterated analogue of methyl phosphite, CH(3)OP((2)H)(O)(2)(-). The electron to (31)P and (2)H nuclear scalar coupling constants were found to be 4.6 and 0.10 MHz, respectively. In contrast, the effects of paramagnetic ions on the methoxy and ethoxy (1)H resonances of MeOPH and MEP are weak, and the evidence suggests that relaxation of these nuclei occurs by a dipolar mechanism. The wide variation in the relaxation sensitivities of the (1)H and (31)P nuclei of MeOPH and MEP permits us to study how differences in the strengths of the interactions between an observed nucleus and a paramagnetic center affect NMR T(2) relaxations. We propose that these anion ligand probes may be used to study ligand-exchange reactivities of manganese complexes without requiring variable temperature studies. The (31)P T(2) is determined by chemical association kinetics when the following condition is met: (T(2M,P)/T(2M,H))(Deltanu(P)/Deltanu(HP) - 1) < 0.2 where T(2M,P) and T(2M,H) are the transverse relaxation times of the (31)P and (1)H nuclei when the probe is bound to the metal, and Deltanu(P) and Deltanu(HP) are the paramagnetic line broadenings of the (31)P and (1)H-P nuclei, respectively. We assert that the ratio T(2M,P)/T(2M,H) can be estimated for a general metal complex using the results of EPR and NMR experiments. PMID- 11735463 TI - Are halocarboranes suitable for substitution reactions? The case for 3-I-1,2 closo-C(2)B(10)H(11): molecular orbital calculations, aryldehalogenation reactions, (11)B NMR interpretation of closo-carboranes, and molecular structures of 1-Ph-3-Br-1,2-closo-C(2)B(10)H(10) and 3-Ph-1,2-closo-C(2)B(10)H(11). AB - In this paper, the chemistry of 3-X-1,2-closo-C(2)B(10)H(11) (X = halogen) derivatives is extended. Molecular orbital and (11)B and (13)C NMR calculations on these species are presented. A qualitative interpretation of the (11)B NMR spectra of closo o-carborane derivatives is also provided. The synthesis of 3-X-1 R-o-carborane (X = I, Br and R = Me, Ph) derivatives is reported, and aryldehalogenation at the B3 position is reported for the first time. The molecular and crystal structures of 1-phenyl-3-bromo-1,2-dicarba-closo dodecaborane and 3-phenyl-1,2-dicarba-closo-dodecaborane are described. PMID- 11735464 TI - Synthesis and reactivity of the monocarbon molybdenacarborane anion [1,2-mu NHBu(t)-2,2,2-(CO)(3)-closo-2,1-MoCB(10)H(10)](-). AB - The reagent Li[7-NHBu(t)()-nido-7-CB(10)H(12)] reacts with [Mo(CO)(6)] in NCMe at reflux temperatures, followed by addition of [N(PPh(3))(2)]Cl, to give [N(PPh(3))(2)][1,2-mu-NHBu(t)-2,2,2-(CO)(3)-closo-2,1-MoCB(10)H(10)] (1). The tungsten (2) and chromium (3) analogues were similarly obtained, but the latter is unstable and was isolated in low yield. An X-ray diffraction study of 2 confirmed that the exo-polyhedral NHBu(t) group forms a bridge between the cage carbon atom and the tungsten. For 1, this intramolecular donor bond is lifted on protonation in the presence of donor molecules L (CO, PPh(3), PMe(3), PEt(3), PMe(2)Ph) when zwitterionic complexes [1-NH(2)Bu(t)()-2,2,2-(CO)(3)-2-L-closo-2,1 MoCB(10)H(10)] (4) are formed. In contrast, protonation with HCl gives a salt [N(PPh(3))(2)][1-NH(2)Bu(t)()-2,2,2-(CO)(3)-2-Cl-closo-2,1-MoCB(10)H(10)] (5). Complex 1 in CH(2)Cl(2) with CNBu(t) is oxidized by iodine, affording [1,2-mu NHBu(t)()-2,2,2-(CNBu(t)())(3)-2-I-closo-2,1-MoCB(10)H(10)] (6a). Treatment of 1 with [CuCl(PPh(3))](4) in the presence of Tl[PF(6)] yields the bimetallic compound [exo-[Cu(PPh(3))]-1,2-mu-NHBu(t)-2,2,2-(CO)(3)-closo-2,1-MoCB(10)H(10)] (8), whereas reaction with [AuCl(PPh(3))] and Tl[PF(6)] affords a mixture of [1,2 mu-NHBu(t)-2-[Au(PPh(3))]-2,2,2-(CO)(3)-closo-2,1-MoCB(10)H(10)] (9) and [Au(PPh(3))(2)][2,2'-mu-Au-[1,2-mu-NHBu(t)-2,2,2-(CO)(3)-closo-2,1 MoCB(1)(0)H(10)](2)] (10a). In solution, 9 disproportionates, giving 10a. The [N(PPh(3))(2)](+) salt (10b) is readily prepared by treating 1 with [AuCl(THT)] (THT = tetrahydrothiophene) and Tl[PF(6)], and its structure was determined by X ray diffraction. PMID- 11735465 TI - Synthesis and NMR studies of new DOTP-like lanthanide(III) complexes containing a hydrophobic substituent on one phosphonate side arm. AB - Three derivatives of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(methylene phosphonic acid) (DOTP) containing a hydrophobic substituent on one side chain were prepared and their lanthanide complexes examined by NMR. The new ligands include 1-(1-octyl-methyl-phosphonic acid)-4,7,10-tris(methylene phosphonic acid) 1,4,7,10-tetraazacyclododecane (C(8)-DOTP), 1-(1-undecyl-methyl-phosphonic acid) 4,7,10-tris(methylene phosphonic acid)-1,4,7,10-tetraazacyclododecane (C(11) DOTP), and 1-(1-4-nitro-phenyl-methyl-phosphonic acid)-4,7,10-tris(methylene phosphonic acid)-1,4,7,10-tetraazacyclododecane (NO(2)-Ph-DOTP). (1)H NMR spectra of the ytterbium(III) complexes were assigned by using a combination of COSY spectroscopy and a fitting procedure that matches experimental NMR hyperfine shifts with those estimated from a MMX-derived structure. The analysis showed that a single isomer is present in solution and that the bulky hydrophobic substituent occupies the less sterically demanding H(6) equatorial position in the YbL(5)(-) complexes. Although the YbL(5)(-) complexes have lower symmetry due to the added substituent, the average (1)H hyperfine shifts are 5-10% larger in these complexes compared to YbDOTP(5)(-). This was magnified further in the hyperfine (23)Na NMR shifts of ion-paired sodium ions where the extracellular Na(+) signal in perfused rat hearts displayed a 28% larger hyperfine shift in the presence of Tm(C(11)-DOTP)(5)(-) than with an equivalent amount of TmDOTP(5)(-). PMID- 11735466 TI - The Gd(3+) complex of a fatty acid analogue of DOTP binds to multiple albumin sites with variable water relaxivities. AB - The 20 MHz water relaxivity (r(1)) of gadolinium(III) complexes formed with two fatty acid analogues of 1,4,7,10-tetraazacyclododecane-1,4,7,10 tetrakis(methylene phosphonate) were shown to increase substantially in the presence of albumin. The r(1) values of Gd(C(8)-DOTP)(5-) and Gd(C(11)-DOTP)(5-) in water were similar to that of the parent GdDOTP(5-), a q = 0 complex known to relax water very efficiently via an outer-sphere mechanism. Neither fatty acid analogue formed apparent aggregates or micelles in water up to 20 mM, but both showed dramatic increases in r(1) upon addition of albumin. Further ultrafiltration studies of Gd(C(11)-DOTP)(5-) in the presence of non-defatted HSA showed that the complex binds at a minimum of five high-affinity fatty acid sites with stepwise binding constants ranging from 1.27 x 10(5) to 2.7 x 10(3) M(-1). The 20 MHz relaxivity of Gd(C(11)-DOTP)(5-) in the presence of excess HSA was 23 mM(-1) s(-1) at 25 degrees C. The NMRD curve showed a broad maximum 20-30 MHz which fitted well to standard theory for a q = 0 complex with rapid outer-sphere water exchange. The r(1b) of Gd(C(11)-DOTP)(5-) bound at the tightest site on HSA was approximately 40 mM(-1) s(-1) at 5 degrees C, an extraordinarily high value for an outer-sphere complex. However, the r(1b) of Gd(C(11)-DOTP)(5-) bound at the weaker sites on HSA was considerably lower, approaching the relaxivity of the free complex in water. This suggests that the complex bound in the highest affinity fatty acid site is less mobile than the same complex bound at the weaker affinity fatty acid sites. This combined ultrafiltration and relaxivity study demonstrates that the common assumption of a single r(1b) value for a Gd(3+) complex bound at several protein sites is not a valid approximation. PMID- 11735467 TI - Group 4 dimethylmetallocenes: improved synthesis and reactivity studies. AB - Group 4 dimethylmetallocenes are catalyst precursors for the methylmetallocenium/borate catalyst systems for olefin polymerization, and they are usually prepared by methylation (with MeMgCl or MeLi) of the parent metallocene dichlorides. We describe here a simpler preparation of a series of bisindenyldimethylmetallocenes carried out by reacting the pi-ligand with a 2 fold excess of MeLi, and then MtCl(4) (Mt = Ti, Zr, Hf). This simple, one-pot method produces the dimethylated complexes in higher overall yield, and saves on reaction time and solvents. Ind(2)MtMe(2) (1, Mt = Ti; 2a, Mt = Zr; 3, Mt = Hf), (4,7-Me(2)Ind)(2)ZrMe(2) (4), rac/meso-[C(2)H(4)(Ind)(2)]ZrMe(2) (5), meso [C(2)H(4)(4,7-Me(2)Ind)(2)]ZrMe(2) (m-6a), and meso-[C(2)H(4)(4,7 Me(2)Ind)(2)]HfMe(2) (m-7a) have been prepared in 40-80% isolated yields. 2a reacts with 1-4 equiv of t-BuOH to give the mono-tert-butoxy derivative 2b, Ind(2)ZrMe(O-t-Bu), while reaction with 2 equiv of C(6)F(5)OH cleanly affords Ind(2)Zr(OC(6)F(5))(2) (2c). Analogously, in the presence of 2 equiv of t-BuOH, m 6a gives meso-[C(2)H(4)(4,7-Me(2)Ind)(2)]ZrMe(O-t-Bu) (m-6b) with replacement of the outward methyl group only, as established by NMR analysis; meso-[C(2)H(4)(4,7 Me(2)Ind)(2)]Zr(OC(6)F(5))(2) (m-6c) is obtained by reaction with 2 equiv of C(6)F(5)OH. The molecular structures of m-6a and m-6c are also described. PMID- 11735468 TI - Preparation, structure, and properties of the corner-shared double cubes [Mo(6)HgQ(8)(H(2)O)(18)](8+) (Q = S, Se) and tungsten analogues. AB - The purple corner-shared double cube [Mo(6)HgS(8)(H(2)O)(18)](8+) derivative of green [Mo(3)S(4)(H(2)O)(9)](4+), obtained under air-free conditions by the reaction with Hg(0) (metal), is also formed with Hg(I)(2). The Hg(I)(2) reaction is accounted for by the disproportionation Hg(I)(2) <==> Hg(0) + Hg(II), which is a source of Hg(0). X-ray crystallographic information on the blue partially Cl(-) substituted cucurbituril supramolecular assemblies [Mo(6)HgQ(8)Cl(4)(H(2)O)(14)](C(36)H(36)N(24)O(12))Cl(4).14H(2)O (1) and of the Se analogue [Mo(6)HgSe(8)Cl(4) (H(2)O)(14)](C(36)H(36)N(24)O(12))Cl(4).14H(2)O (2) have been determined. The product [W(6)HgSe(8)Cl(4)(H(2)O)(14)](C(36)H(36)N(24) O(12)) Cl(4).14H(2)O (3) has also been obtained, but there is no evidence for [W(6)HgS(8)(H(2)O)(18)](8+) and related forms. The formation of [Mo(6)HgS(8)(H(2)O)(18)](8+) by the reaction of [Mo(3)S(4) (H(2)O)(9)](4+) with Hg(0) under anaerobic conditions maximizes after approximately 40 h in 2.0 M HCl, but requires longer reaction time ( approximately 120 h) in 2.0 M Hpts (p-toluenesulfonic acid) and in 2 M HClO(4) ( approximately 6 days). In 2.0 M HCl there is little absorbance increase until [Mo(3)S(4)(H(2)O)(9)](4+) exceeds 1.2 x 10(-)(3) M, which is explained by a dependence of the formation K (265 M(-1)) on [Mo(3)S(4)(H(2)O)(9)(4+)](2). Furthermore, on dilution of column-purified [Mo(6)HgS(8)(H(2)O)(18)](8+), Beer's law is not obeyed and equilibria involving 2[Mo(3)S(4)(H(2)O)(9)](4+) are apparent. The kinetics of formation of [Mo(6)HgS(8)(H(2)O)(18)](8+) is first order in [Mo(3)S(4)(H(2)O)(9)](4+), consistent with rate-determining formation of the single cube [Mo(3)HgS(4)(H(2)O)(x)](4+). The oxidations of [Mo(6)HgS(8)(H(2)O)(18)](8+) with [Fe(H(2)O)(6)](3+) and [Co(dipic)(2)](-) are complicated by the release of [Hg(H(2)O)(6)](2+), which also functions as an oxidant. Similar results are obtained for [Mo(6)HgSe(8)(H(2)O)(18)](8+) and the less extensively studied [W(6)HgSe(8)(H(2)O)(18)](8+). PMID- 11735469 TI - Modulation of coordination chemistry in copper(I) complexes supported by Bis[2-(2 pyridyl)ethyl]amine-based tridentate ligands. AB - Structure and physicochemical properties of copper(I) complexes of the tridentate ligands L(2) (N,N-bis[2-(6-methylpyridin-2-yl)ethyl]phenethylamine) and L(3) (N,N bis[2-(2-pyridyl)ethyl]-beta-methylphenethylamine) have been examined to obtain deeper insights into modulation of the coordination chemistry of copper(I) complexes. [Cu(I)(L(2))(CH(3)CN)](ClO(4)) (2.CH(3)CN) has a distorted tetrahedral geometry, which consists of three nitrogen atoms of the ligand and one nitrogen atom of the bound CH(3)CN. Steric repulsion between the 6-methyl group on the pyridine nucleus of L(2) and the metal ion of the complex prevents the cuprous complex from adaptation to a three-coordinate geometry which must have a shorter Cu-N(pyridine) distance ( approximately 1.88 A). Thus, the four-coordinate copper(I) complex (2.CH(3)CN) with a longer Cu-N bond (1.98 approximately 2.13 A) becomes favorable, resulting in rather strong binding of CH(3)CN to the metal ion. In [Cu(I)(L(3))](ClO(4)) (3), there is a Cu(I)-pi interaction between the cuprous ion and the phenyl group of the ligand sidearm. Such a copper(I)-arene interaction is essentially weak, but is significantly stabilized in complex 3. The methyl group at the benzylic position of L(3)() reduces the degree of freedom of sidearm rotation to make the phenyl group stick on the cuprous ion. Thus, the reactivity of the copper(I) complexes of L(2) and L(3) toward dioxygen is significantly diminished, showing sharp contrast to the high reactivity of the copper(I) complex supported by a similar tridentate ligand L(1) (N,N-bis[2-(2 pyridiyl)ethyl]phenethylamine). PMID- 11735470 TI - Facile C(sp(2))/OR bond cleavage by Ru or Os. AB - Os(H)(3)ClL(2) (L = P(i)Pr(3) or P(t)Bu(2)Me) are shown to be useful "precursors" to "OsHClL(2)", which react with vinyl ethers to form first an eta(2)-olefin adduct and then isomerize to the carbenes, OsHCl[CMe(OR)]L(2). Subsequent R- and L-dependent reactions involve C(sp(2))-OR bond cleavage, to make either carbyne or vinylidene complexes. The mechanisms of these reactions are explored, and the thermodynamic disparity of Ru versus Os and the influence of the OR group and the spectator phosphine ligands are discussed based on DFT (B3PW91) calculations. PMID- 11735471 TI - Carborane-containing liquid crystals: synthesis and structural, conformational, thermal, and spectroscopic characterization of diheptyl and diheptynyl derivatives of p-carboranes. AB - Molecular and electronic structures for four p-carborane derivatives were studied in the context of their liquid crystalline properties. Thus molecular and crystal structures of diheptyl and diheptynyl derivatives of 10- and 12-vertex bi-p carboranes were determined by X-ray crystallography and compared to the results of ab initio calculations at the HF/6-31G level of theory. Experimentally observed significant positional disorder of one of the substituents in the 10 vertex derivatives, 2[2]a and 2[2]b, was related to conformational properties of the alkyl-carborane bond. Experimental and theoretical studies of the electronic structures were conducted for the four compounds using UV and NMR spectroscopies. The nature of the unique long wavelength absorption band at 232 nm in the diheptynyl derivative 2[2]b was explained using INDO/2//HF/6-31G analysis. The complete assignment of the (13)C signals was accomplished using a long-range coupling technique and was supported by the calculated (HF/6-31G) isotropic shielding tensors. Analysis of absorption spectra, NMR substituent effects, and trends in bond lengths shows generally strong cage-acetylene electronic interactions for the 10-vertex p-carborane, while the 12-vertex p-carborane remains largely electronically isolated. Ab initio calculations revealed that 12 vertex p-carborane has significantly larger electronic polarizability and quadrupole moments than the 10-vertex analogues, which are larger than those for bicyclo[2.2.2]octane compounds. All these results on packing, conformational, and electronic properties form the basis for the discussion of thermal behavior of the four carborane compounds, bicyclo[2.2.2]octane analogues, and some related compounds. PMID- 11735472 TI - Magnetic-field-dependent heat capacity of the single-molecule magnet [Mn(12)O(12)(O(2)CEt)(16)(H(2)O)(3)]. AB - Accurate heat capacities of the single-molecule magnet [Mn(12)O(12)(O(2)CEt)(16)(H(2)O)(3)] were measured from 0.3 to 311 K by adiabatic calorimetry without an external magnetic field. Heat-capacity anomalies were separated by assuming several contributions including lattice vibration, magnetic anisotropy, and hyperfine splitting. Among them, a tiny thermal anomaly between 1 and 2 K is attributable to the presence of Jahn-Teller isomers. The heat capacities of the polycrystalline sample were also measured with applied magnetic fields from 0 to 9 T in the 2-20 K temperature region by the relaxation method. With an applied magnetic field of up to 2 T, a steplike heat-capacity anomaly was observed around the blocking temperature T(B) approximately 3.5 K. The magnitude of the anomaly reached a maximum at 0.7 T. With a further increase in the magnetic field, the step was decreasing, and finally it disappeared above 3 T. The step at T(B) under 0.7 T can be roughly accounted for by assuming that a conversion between the up-spin and down-spin states is allowed above T(B) by phonon-assisted quantum tunneling, while it is less effective below T(B). Excess heat capacity under a magnetic field revealed a large heat-capacity hump around 14 K and 2 T, which would be attributed to a thermal excitation from the S = 9 ground state to the spin manifold with different S values, where S is the total spin quantum number. PMID- 11735473 TI - Shaping the cavity of the macrocyclic ligand in metallocalix[4]arenes: the role of the ligand sphere. AB - The coordination form of calix[4]arene ligands and therefore the cavity of the macrocyclic ligand can be controlled by other ligands in transition metal calix[4]arene complexes, if strong directing coligands such as oxo groups are used. This paper describes the synthesis and characterization of the d(0) transition metal complexes [Cax(OMe)(2)O(2)TiCl(2)] 1 (monoclinic, space group P2(1)/c, lattice constants a = 21.639(4), b = 20.152(3), c = 12.750(3) A, beta = 95.68(3), V = 5532.6(19) A(3)) and [Cax(OMe)(2)O(2)MoO(2)] 2 (monoclinic, space group P2/c, lattice constants a = 12.433(3), b = 16.348(3), c = 24.774(5) A, beta = 99.15(3), V = 4971.6(17) A(3)). Whereas in 1 the calix[4]arene ligand adopts an elliptically distorted cone conformation, the macrocyclic ligand binds in a paco like conformation to the metal center of 2, in the solid state and in solution. This was predicted by density functional theory calculations on models of different isomers of 1 and 2: cis,cone-1',2', trans,cone-1',2', and cis,paco 1',2'. According to these calculations, the energetic difference of 72.9 kJ/mol between both cis-dioxomolybdenum compounds is quite pronounced in favor of the cis,paco isomer, and 28.0 kJ/mol for the titanium compounds in favor of the cis,cone isomer. PMID- 11735474 TI - Regular and inverse secondary kinetic enthalpy effects (KHE) for the rate of inversion of thioether and 1,1'-biisoquinoline complexes of ruthenium and osmium. AB - Thioether complexes with the formula Delta/Lambda-chloro(thioether)bis(2,2' bipyridine)metal(II) (M = Ru, Os; thioether = dimethyl sulfide (3a(+)), diethyl sulfide (3b(+)), and tetrahydrothiophene (3c(+))) have been synthesized. The rates of inversion at the sulfur atom of the thioether ligands have been measured by spin-inversion transfer and line-shape NMR methods. In every case, the ruthenium derivative exhibits a faster inversion frequency at a given temperature than the corresponding osmium derivative. In contrast, similar complexes with the formula chloro(delta/lambda-1,1'-biisoquinoline)(2,2':6',2" terpyridine)metal(II), 4(M=Ru,Os)(+), undergo atropisomerization of the misdirected 1,1'-biisoquinoline (1,1'-biiq) ligand with rates that are faster for osmium than ruthenium. As a result of the lanthanide contraction effect and the similar metric parameters associated with the structures of second-row and third row transition metal derivatives, steric factors associated with the isomerizations are presumably similar for the Ru and Os derivatives of these compounds. Since third-row transition metal complexes tend to have larger bond dissociation enthalpies (BDE) than their second-row congeners, we conclude the difference in reactivities of 3(M=Ru)(+) versus 3(M=Os)(+) and 4(M=Ru)(+) versus 4(M=Os)(+) are attributed to electronic effects. For 3, the S3p lone pair of the thioether, the principal sigma donor orbital, is orthogonal to the metal sigma acceptor orbital in the transition state of inversion at sulfur and the S 3s orbital is an ineffective sigma donor. Thus, a regular relationship between the kinetic and thermodynamic stabilities of 3(M=Ru)(+) and 3(M=Os)(+) is observed for the directed <==> [misdirected] <==> directed (DMD) isomerization (the more thermodynamically stable bond is less reactive). In contrast, atropisomerization of 4(+) involves redirecting (strengthening) the M-N bonds of the misdirected 1,1'-biiq ligand in the transition state. Therefore, an inverse relationship between the kinetic and thermodynamic stabilities of 4(M=Ru)(+) and 4(M=Os)(+) is observed for the misdirected <==> [directed] <==> misdirected (MDM) isomerization (the more thermodynamically stable bond is more reactive). The rates obtained for 4(+) are consistent with the rates of atropisomerization of Delta/Lambda (delta/lambda-1,1'-biisoquinoline)bis(2,2'-bipyridine)metal(II), 1(M=Ru,Os)(2+), and (eta(6)-benzene) Delta/Lambda-(delta/lambda-1,1' biisoquinoline)halometal(II), 2(M=Ru,Os;halo=Cl,I)(+), that we reported previously. We term the relative rates of reaction of second-row versus third-row transition metal derivatives kinetic element effects (KEE = k(second)/k(third)). While the KEE appears to be generally useful when comparing reactions of isostructural species (e.g. the relative rates of 1(M=Ru)(2+), 1(M=Os)(2+), and 1(M=Ir)(3+)), different temperature dependencies of reactions prevent the comparison of related reactions between species that have different structures (e.g., the 1,1'-biiq atropisomerization reactions of 1(M=Ru,Os)(2+) versus 2(M=Ru,Os;halo=Cl,I)(+) versus 4(M=Ru,Os)(+)). This problem is overcome by comparing entropies of activation and kinetic enthalpy effects (KHE = DeltaH(third)/DeltaH(second)). For a given class of 1,1'-biiq complexes, we observe a structure/reactivity relationship between DeltaH and the torsional twist of the 1,1'-biiq ligands that are measured in the solid state. PMID- 11735475 TI - Syntheses and physical properties of quasi-one-dimensional halogen-bridged Cu(II)(-)Pt(IV) mixed-metal complexes [Cu(chxn)(2)][PtX(2)(chxn)(2)]X(4). AB - Quasi-one-dimensional halogen-bridged Cu(II)-Pt(IV) mixed-metal complexes of the form [Cu(chxn)(2)][PtX(2)(chxn)(2)]X(4), where chxn = 1R,2R-diaminocyclohexane and X is either Cl or Br, have been synthesized. The crystal structures of these compounds have been determined by single-crystal X-ray diffraction. The Cl bridged compound crystallizes in the space group I222 with dimensions a = 24.237(1) A, b = 5.103(1) A, c = 6.854(1) A, and V = 847.7(1) A(3) and with Z = 1. The Br-bridged complex crystallizes in the space group I222 with dimensions a = 23.700(8) A, b = 5.344(5) A, c = 6.978(8) A, and V = 883.8(8) A(3) and with Z = 1. These structures are isomorphic to each other and to homometal [Pt(chxn)(2)][PtX(2)(chxn)(2)]X(4) complexes. In these complexes, the planar [Cu(chxn)(2)] and the octahedral [PtX(2)(chxn)(2)] groups are stacked alternatively with the axial bridging halogen ions, forming linear chain structures. The neighboring [Cu(chxn)(2)] and [PtX(2)(chxn)(2)] moieties along the chains are linked by hydrogen bonds between amino hydrogens and the counteranions (X). Moreover, there are hydrogen bonds among the neighboring chains that form a two-dimensional hydrogen-bonded network parallel to the bc plane. Therefore, the Cu(II) and Pt(IV) units are two-dimensionally ordered. The b axes correspond to the Cu(II)-Pt(IV) separations, which are shorter than those of [Pt(chxn)(2)][PtX(2)(chxn)(2)]X(4) due to the smaller ionic radius of the Cu(II) ions. In the XP spectra, the Pt(IV) 4f(7/2) and Pt(IV) 4f(5/2) binding energies in homometal [Pt(chxn)(2)][PtX(2)(chxn)(2)]X(4) are lower than those of [Cu(chxn)(2)][PtX(2)(chxn)(2)]X(4) (X = Cl and Br), indicating that the electron phonon interaction in Cu(II)-Pt(IV) compounds is stronger than that in Pt(II) Pt(IV) compounds. In the Raman spectra, nu(Pt(IV)(-)X) of the homometal Pt(II) Pt(IV) complexes is lower than that of the Cu(II)-Pt(IV) complexes, indicating again that the electron-phonon interaction in Cu(II)-Pt(IV) compounds is stronger than that of Pt(II)-Pt(IV) compounds. The temperature-dependent magnetic susceptibilities of the Cu(II)-Pt(IV) complexes show weak antiferromagnetic interactions between Cu(II) components along the chain axes. PMID- 11735476 TI - Tris(pyridinealdoximato)metal complexes as ligands for the synthesis of asymmetric heterodinuclear Cr(III)M species [M = Zn(II), Cu(II), Ni(II), Fe(II), Mn(II), Cr(II), Co(III)]: a magneto-structural study. AB - Reactions of the LCr(III) unit with an in situ prepared M(PyA)(3)(n-) ion, where L represents 1,4,7-trimethyl-1,4,7-triazacyclononane and PyA(-) is the monoanion of pyridine-2-aldoxime, yield heterodinuclear complexes of general formula [LCr(III)(PyA)(3)M](2+/3+) as perchlorate salts, where M = Cr(II) (1), Mn(II) (2), low-spin Fe(II) (3), Ni(II) (4), Cu(II) (5), Zn(II) (6), and low-spin Co(III) (7). These compounds contain three oximato anions as bridging ligands. The hexadentate ligand with the identical donor atoms, tris(2-aldoximato-6 pyridyl)phosphine, P(PyA)(3), has been employed to prepare a second Cr(III)Ni(II) species 8, whose magnetic properties differ significantly from those of 4. Complexes 1-8 have been characterized on the basis of elemental analysis, mass spectrometry, IR, UV-vis, Mossbauer, and EPR spectroscopies, and variable temperature (2-295 K) magnetic susceptibility measurements. They are isostructural in the sense that they all contain a terminal Cr(III) ion in a distorted octahedral environment, CrN(3)O(3), and a second six-coordinated metal ion M in a mostly trigonal prismatic MN(6) geometry. The crystal structures of the perchlorate salts of 2-5, 7, and 8 have been determined by X-ray crystallography at 100 K. The structures consist of mixed-metal Cr(III)M(II) and Cr(III)Co(III) complexes with a geometry in which two pseudooctahedral polyhedra are joined by three oximato (=N-O(-)) groups, with an intramolecular Cr.M(Co) distance in the range of 3.4-3.7 A. The cyclic voltammograms of the complexes reveal ligand oxidation and reduction processes, and in addition, metal-centered oxidation processes have been observed. X-band EPR spectroscopy has been used to establish the electronic ground state of the heterodinuclear complexes. Analysis of the susceptibility data indicates the presence of weak exchange interactions, both ferro- and antiferromagnetic, between the paramagnetic centers. A qualitative rationale on the basis of the Goodenough-Kanamori rules is provided for the difference in magnetic behaviors. PMID- 11735477 TI - Formation, isomerization, and derivatization of Keggin tungstoaluminates. AB - Trends in the stability of alpha- and beta-Keggin heteropolytungstates of the second-row main-group heteroatoms Al(III), Si(IV), and P(V) are elaborated by data that establish the roles of kinetic and thermodynamic control in the formation and isomerization of Keggin tungstoaluminates. Slow, room-temperature co-condensation of Al(III) and W(VI) (2:11 molar ratio) in water gives a pH 7 solution containing beta(1) and beta(2) isomers of [Al(AlOH(2))W(11)O(39)](6)(-) (beta(1)- and beta(2)-1). Partial equilibration of this kinetic product mixture by gentle heating (2 h at 100 degrees C) or, alternatively, co-condensation of Al(III) and W(VI) for 2.5 h at 100 degrees C both give mixtures of beta(2)-, beta(3)-, and alpha-1. Full equilibration, by prolonged heating (25 days at 100 degrees C), gives an isomerically pure solution of alpha-1, thus demonstrating that isomerization occurs in the direction beta(1) --> beta(2) --> beta(3) --> alpha. Furthermore, kinetically controlled conversions of 1 to H(5)[AlW(12)O(40)] (2)-achieved by heating pH 0-0.2 solutions of 1 for 5 days at 100 degrees C-occur with retention of isomeric integrity, such that alpha-1 is converted to alpha-2 (92%; 8% beta), while mixtures of beta(2)- and beta(3)-1 are converted to beta-2 (87%; 13% alpha). These data, when combined with previously reported observations (equilibria between alpha- and beta-2, kinetically controlled hydrolyses of alpha 2 to alpha-[AlW(11)O(39)](9)(-) (alpha-3) and of beta-2 to beta(2)-3, and equilibria between beta(3)- and alpha-3), provide a comprehensive picture regarding the roles of kinetic and thermodynamic control. Finally, a general method for preparation of the isomerically pure derivatives alpha-K(9)( )(n)()[AlM(n)()(+)W(11)O(39)] (4), M(n)()(+) = Al(III), [V(IV)O](2+), [V(V)O](3+), Mn(II), Mn(III), Mn(IV), Co(II), and Co(III), is provided. The presence of Mn(IV) is confirmed by cyclic voltammetry, pK(a) values of the aquo ligands on 4 are determined by pH titration, and the isomeric structure of these derivatives is established by (27)Al, (51)V, and (183)W NMR and IR spectroscopies and X-ray crystallography. PMID- 11735478 TI - Synthesis and cleavage reactions of metal-metal-bonded [Mo(2)(S(2)CNR(2))(6)](OTf)(2), a source of the Tris(dithiocarbamato)molybdenum(IV) fragment. AB - Halide abstraction from the chlorotris(dialkyldithiocarbamato)molybdenum(IV) complexes MoCl(S(2)CNR(2))(3) (R = Et, Me) with silver triflate produces the diamagnetic dimeric complexes [Mo(2)(S(2)CNR(2))(6)](OTf)(2) in good yield. The crystallographically determined structure of the diethyldithiocarbamato complex indicates that the dimer consists of two pentagonal bipyramids sharing an axial edge, with a Mo-Mo separation (2.8462(8) A) indicative of a metal-metal bond. A qualitative analysis of the bonding indicates that this bond is of order 2 and consists of one normal sigma bond and one relatively weak "skewed pi" interaction. The dimers [Mo(2)(S(2)CNR(2))(6)](OTf)(2) react with a variety of reagents to give monomeric seven-coordinate complexes, including the new cationic molybdenum(IV) complex [Mo(PMe(2)Ph)(S(2)CNEt(2))(3)](OTf), which has been structurally characterized. Kinetic studies of the reaction of [Mo(2)(S(2)CNEt(2))(6)](OTf)(2) with halides indicate the presence of competing dissociative and associative substitution pathways, although neutral donors may react by different mechanisms. PMID- 11735479 TI - Crystal and solution structures of N,N-dimethylthioformamide-solvated copper(I), silver(I), and gold(I) ions studied by X-ray diffraction, X-ray absorption, and vibrational spectroscopy. AB - Crystal structures of the solvated copper(I) and silver(I) perchlorate salts crystallizing from N,N-dimethylthioformamide solution have been determined by single-crystal X-ray diffraction at 295 K. Tetrakis(N,N dimethylthioformamide)copper(I) perchlorate, [Cu(SCHN(CH(3))(2))(4)]ClO(4), crystallizes in the monoclinic space group P2/n (No. 13) with a = 8.428(2), b = 9.605(2), and c = 15.096(3) A, beta = 104.35(2) degrees, and Z = 2. The copper(I) ion in a site of C(2) symmetry coordinates four N,N-dimethylthioformamide ligands in a slightly distorted tetrahedral coordination with Cu-S bond distances of 2.3249(8) and 2.3494(8) A. The triclinic (P1, No. 2) tris(N,N dimethylthioformamide)silver(I) perchlorate, Ag(SCHN(CH(3))(2))(3)ClO(4), with a = 7.4149(5), b = 7.7953(5), and c = 17.1482(1) A, alpha = 98.341(5), beta = 93.910(5), and gamma = 107.084(5) degrees, and Z = 2, contains centrosymmetric Ag(2)(SCHN(CH(3))(2))(6)(2+) dimers in which two almost planar AgS(3) units are held together by an asymmetric double sulfur bridge with one short and one long Ag-S bond, 2.529(1) and 2.930(1) A, respectively. The Ag-S bond distances to the two terminal N,N-dimethylthioformamide ligands are 2.469(1) and 2.543(1) A. The solvated copper(I) and silver(I) ions in solution were found by means of large angle X-ray scattering (LAXS) to coordinate four N,N-dimethylthioformamide molecules with the mean Cu-S and Ag-S bond distances 2.36(1) and 2.58(1) A, respectively, probably with distorted tetrahedral coordination geometry, while an EXAFS study gave the Cu-S bond distance 2.34(1) A. EXAFS studies showed a linear S-Au-S entity with an Au-S bond distance of 2.290(5) A in the structure of the solid bis(N,N-dimethylthioformamide)gold(I) tetrafluoroborate, Au(SCHN(CH(3))(2))(2)BF(4). The structure in solution is similar with a mean Au-S bond distance of 2.283(4) A. Raman and infrared vibrational spectra of the solvated copper(I), silver(I), and gold(I) ions in the solid state and N,N dimethylthioformamide solution have been recorded and assigned. PMID- 11735480 TI - New pillared layered gallium phosphonates in the gallium/1,2-ethylenediphosphonic acid system. AB - 1,2-Ethylenediphosphonic acid reacts with gallium nitrate in water to give new pillared layered gallium phosphonates, the structure of which depends on the experimental conditions used for their preparation. Thus, Ga(4)(O(3)PC(2)H(4)PO(3))(3) (1) and Ga(2)(OH)(2)(O(3)PC(2)H(4)PO(3)) (2) were isolated and the structure of 1 was solved by single-crystal X-ray diffraction, while 2 was shown to have the same metal/PO(3) arrangement as the previously described Ga(OH)(O(3)PCH(3)). Ga(4)(O(3)PC(2)H(4)PO(3))(3) (1) is triclinic, with space group P1 with Z = 2, a = 5.1480(4), b = 8.0354(7), and c = 12.383(1) A; alpha = 91.34(1), beta = 101.40(1), and gamma = 90.86(1) degrees; V = 501.9(1) A(3). The structure of 1 is unusual with (i) mixed GaO(4) and GaO(5) sites while a 6-fold coordination is observed for gallium in (2) and (ii) the presence of the organic moiety within both the layers and the interlayer space. The two compounds were fully characterized using (31)P and (71)Ga solid-state NMR. PMID- 11735481 TI - Palladium(II) and platinum(II) analogues of luminescent diimine Triangulo complexes supported by triply bridging sulfide ligands: structural and spectroscopic comparisons. AB - Reaction of [M(L-L)Cl(2)] [M = Pd, Pt; L-L = 4,4'-di-tert-butyl-2,2'-bipyridine ((t)Bu(2)bpy), 4,4'-dimethylcarboxylate-2,2'-bipyridine ((CO(2)Me)(2)bpy), bis(diphenylphosphino)methane (dppm)] with Na(2)S in refluxing methanol afforded [M(3)((t)Bu(2)bpy)(3)(mu(3)-S)(2)](2+) [M = Pd (1a), Pt (2a)], [M(3)((CO(2)Me)(2)bpy)(3)(mu(3)-S)(2)](2+) [M = Pd (1b), Pt (2b)], and [Pt(3)(dppm)(3)(mu(3)-S)(2)](2+) (3) as perchlorate salts. X-ray crystal analysis revealed that 1a, 1b, 2a, and 3 have triangular M(3)S(2) core structures. The three metal atoms in 1a, 2a, and 3 form virtual equilateral triangles with intramolecular Pd-Pd and Pt-Pt separations of 3.027(1)-3.065(1) and 3.104(1) 3.154(1) A, respectively. An isosceles triangle of Pd(3) atoms is observed in the molecular structure of 1b. The (1)MLCT absorption of 2a and 2b appears at 415 and 448 nm, respectively, in dichloromethane and is significantly red-shifted from the lowest energy absorption band of the Pd(3) analogues. Complex 1a exhibits weak photoluminescence in the solid state at 77 and 298 K (uncorrected lambda(max) 760 and 730 nm, respectively) while the 77 K solid-state emission of 1b (uncorrected lambda(max) 760 nm) is also weak. At 77 K, complexes 2a, 2b, and 3 display broad unstructured emissions at lambda(max) 616-630 nm in the solid state. Ligand-field excited states are tentatively assigned for these emissions. PMID- 11735482 TI - Bioorganometallic chemistry. 13. Regioselective reduction of NAD(+) models, 1 benzylnicotinamde triflate and beta-nicotinamide ribose-5'-methyl phosphate, with in situ generated [CpRh(Bpy)H](+): structure-activity relationships, kinetics, and mechanistic aspects in the formation of the 1,4-NADH derivatives. AB - Cofactor regeneration; i.e., regiospecific conversion of NAD(+) to 1,4-NADH, has been extensively studied and is a crucial component in the eventual use of 1,4 NADH in a variety of bioorganic synthesis processes involving the formation of chiral organic compounds. We have studied the reduction of a model NAD(+) compound, 1-benzylnicotinamide triflate, 1a, using [CpRh(bpy)(H(2)O)](2+), 2 (Cp = eta(5)-C(5)Me(5), bpy = 2,2'-bipyridyl), as the catalyst precursor and sodium formate (HCO(2)Na) as the hydride source in 1:1 H(2)O/THF and have found exclusive 1-benzyl-1,4-dihydronicotinamide regioselectivity, as was observed previously for natural NAD(+) that provided 1,4-NADH (see: Steckhan et al. Organometallics 1991, 10, 1568). Moreover, a variety of 3-substituted derivatives of 1-benzylpyridinium triflate, in addition to the -C(O)NH(2) group (1a), were also studied to ascertain that this 3-functionality (e.g., -C(O)NHCH(3), C(S)NH(2), -C(O)CH(3), -C(O)OCH(3), and -CN, 1b,d-g) coordinates to a [CpRh(bpy)H](+) complex to direct the concerted, regioselective transfer of the hydride group from the rhodium to the 4-ring position of the NAD(+) model; all coordinating 3-substituents had relative rates in the 0.9-1.3 range with substrate 1a set to 1.0. If in fact the 3-substituent presented a steric effect [ C(O)NH(CH(2)CH(3))(2), 1c] or was a nonbinding group (-CH(3), 1h; -H, 1i), no catalytic hydride transfer was observed even with the more electrophilic 2 and 6 ring positions being readily available, which further implicated the crucial coordination of the NAD(+) model to the CpRh metal ion center. We also found that the 1-benzyl substituent on the nitrogen atom exerted a substantial electron withdrawing effect, in comparison to the electron-donating 1-methyl substituent, and favorably affected the rate of the regioselective reduction (rate enhancement of 1-benzyl/1-methyl = 2.0). The kinetics of the regioselective reduction of 1a were studied to show that the initial rate of reduction, r(i), is affected by the concentrations of the substrate, 1a, precatalyst, 2, and the hydride source, HCO(2)Na, in 1:1 H(2)O/THF: d[1-benzyl-1,4-dihydronicotnamide]/dt = k(cat)[1a][2][HCO(2)Na]. Furthermore, we wish to demonstrate that a previously synthesized aqueous NAD(+) model, beta-nicotinamide ribose-5'-methyl phosphate, 3, shows a similar regioselectivity for the 1,4-NADH analogue, while the initial rate (r(i)) for the regioselective reduction of 3 and NAD(+) itself was found to be comparable in water but faster by a factor of approximately 3 in comparison to 1a in 1:1 H(2)O/THF; the solvent, THF, appeared to inhibit the rate of reduction in 1a by presumably competing with the substrate 1a for the CpRh metal ion center. However, in H(2)O, the initial kinetic rate for substrate 3 was not affected by its concentration and implies that, in H(2)O, [CpRh(bpy)H](+) formation is rate determining. We assume that binding of 3 and NAD(+) to the CpRh metal ion center is also a pertinent step for 1,4-dihydro product formation, the experimental rate expression in H(2)O being d[1,4-dihydro-beta-nicotinamide ribose-5'-methyl phosphate]/dt = k(cat)[2][HCO(2)Na]. What we have discovered, for the first time, is evidence that the regioselective reduction of NAD(+) to 1,4-NADH by [CpRh(bpy)H](+) is a consequence of the amide's ability to coordinate to the CpRh metal center, thereby constricting the kinetically favorable six membered ring transition state for plausible concerted hydride transfer/insertion to C4 to regioselectively provide the 1,4-NADH derivative; [CpRh(bpy)H](+) can be categorized as a biomimetic enzymatic hydride via its ability to bind and regioselectively transfer hydride to C4, exclusively. Clearly, the pyrophosphate and adenosine groups associated with the structure of NAD(+) are not essential in the rate of hydride transfer to C4, with NAD(+) model 3 having a similar initial rate (r(i)) of reduction as NAD(+) itself in water. Finally, a catalytic cycle will be proposed to account for our overall observations. PMID- 11735483 TI - Isomer formation and other issues in the substitution reactions of oxorhenium(V) complexes of 2,2'-bipyridine and related ligands. AB - Two new oxorhenium(V) compounds were prepared and characterized: MeReO(mtp)(Me(2)Bpy) and MeReO(mtp)(dppb), where mtpH(2) is 2 (mercaptomethyl)thiophenol, Me(2)Bpy is 4,4'-dimethyl-2,2'-bipyridine, and dppb is 1,2-(Ph(2)P)(2)C(6)H(4). The more stable geometric isomer of MeReO(mtp)X forms MeReO(mtp)Y (X, Y = PR(3), NC(5)H(4)R) in two steps, both of which show a first order dependence on [Y], proceeding through the metastable geometric isomer MeReO(mtp)Y. When Y = PR(3), no MeReO(mtp)Y was detected at equilibrium; with NC(5)H(4)R, however, both isomers were detected. The values of K(PyPy) were 8.5 9.8, largely irrespective of R; for NC(5)H(5), DeltaH degrees = -4.47 +/- 0.29 kJ and DeltaS degrees = 3.9 +/- 1.0 J K(-1). For the more symmetric edt ligand, geometric isomers do not exist, but enantiomers do. The rate of racemization of MeReO(edt)(NC(5)H(4)R) was proportional to [Py]. Values of k(rac) for 16 compounds span the range 135-370 L mol(-1) s(-1) in C(6)H(6) at 25 degrees C (rho = -0.39 +/- 0.07). In toluene-d(8), k(rac) for 4-picoline has DeltaH = 28.9 +/- 0.4 kJ, DeltaS() = -103.6 +/- 0.9 J K(-1). A common mechanism applies to ligand substitution (mtp) and racemization (edt). MeReO(dithiolate)Py complexes react with Bpy, Me(2)Bpy, Phen, and Me(2)Phen to form six-coordinate chelates, with rate constants 0.024-0.74 L mol(-1) s(-1) at 25 degrees C, some 10(3) times smaller than with pyridines, no doubt owing to the bulk of the bidentates. Values of DeltaS are -86 to -138 J K(-1), reflecting substantial orientational barriers as well as the inherent contribution of the associative mechanism. The product is MeReO(mtp)(Me(2)Bpy). The formation of the metastable isomer is consistent with the mechanism assigned to the ligand substitution and racemization reactions. Such compounds, once formed, no longer participate in ligand substitution reactions at reasonable rates. The formation of the metastable isomer is consistent with the mechanism assigned to the ligand substitution and racemization reactions. PMID- 11735484 TI - Synthesis of arene-soluble mixed-metal uranium/zirconium complexes using the dizirconium nonaisopropoxide ligand. AB - The utility of polydentate monoanionic [Zr(2)(O(i)Pr)(9)](-) (dzni) in generating arene-soluble, mixed-metal Zr/U complexes is described. K[Zr(2)(O(i)Pr)(9)] reacts readily with UI(3)(THF)(4) to form [Zr(2)(O(i)Pr)(9)]UI(2)(THF), 1, in >90% yield. The integrity of the [[Zr(2)(O(i)Pr)(9)]U](2+) unit in 1 was examined by the reaction of 1 with K(2)C(8)H(8), which formed the organometallic complex [Zr(2)(O(i)Pr)(9)]U(C(8)H(8)), 2. In contrast, the reaction of K[Zr(2)(O(i)Pr)(9)] with UCl(4) did not form U(IV)-dzni complexes, and only the ligand exchange product, [UCl(2)(O(i)Pr)(2)(DME)](2), 3, was isolated. The effect of the dzni ligand on the electrochemistry and near-infrared spectroscopy of U(III) is also described. PMID- 11735485 TI - Synthesis and characterization of mixed ruthenium/platinum mu(4)-phosphinidene, phosphorus monoxide, and related clusters. AB - The mixed-metal cluster complexes [Ru(4)(CO)(12)Pt(CO)PPh(3)(mu(4)-PR)] [R = N(i)Pr(2) (1), F (3)] were formed by capping the Ru(3)P face of the nido clusters [Ru(4)(CO)(13)(mu(3)-PR)] with the labile Pt(0) reagent [(eta(2) C(2)H(4))Pt(PPh(3))(2)]. The aminophosphinidene complex 1 undergoes acid hydrolysis to yield the PO complex [Ru(4)(CO)(12)Pt(CO)PPh(3)(mu(4) PO)][H(2)N(i)Pr(2)] (4). The fluorophosphinidene cluster 3 reacts with ethanol to form the alkoxyphosphinidene complex [Ru(4)(CO)(12)Pt(CO)PPh(3)(mu(4)-POEt)] (5). Comparison of spectroscopic and structural data for clusters 1, 3, 4, and 5 reveals the remarkable effects of the mu(4)-phosphinidene and phosphorus monoxide ligands on cluster bonding. PMID- 11735486 TI - Solid-state and solution properties of the lanthanide complexes of a new heptadentate tripodal ligand: a route to gadolinium complexes with an improved relaxation efficiency. AB - The tripodal ligand (alpha,alpha',alpha' 'nitrilotri(6-methyl-2 pyridinecarboxylic acid)) (H(3)tpaa) forms a Gd(III) complex which has a relaxivity (r(1p) = 13.3 mM(-1) s(-1) at 25 degrees C and at 60 MHz) remarkably higher than those of the currently clinically used contrast agents based on octacoordinate polyaminocarboxylate complexes (3.5-4.7 mM(-1) s(-1)) and a reasonably good thermodynamic stability. The crystal structure of the ligand and of its La, Nd, Eu, Gd, Tb, Ho, Tm, Yb, and Lu complexes have been determined by X ray crystallography. The neutral H(3)tpaa molecule adopts, in the solid state, a preorganized tripodal conformation in which the three H(3)tpaa arms are located on the same side of the molecule, ready to bind a metal ion in a heptadentate coordination mode. The structures of the Ln(III) complexes vary along the series for their nuclearity and number of water molecules coordinated to the metal, and a tetrameric structure is observed for the La(3+) ion (9- and 10-coordinate metal centers), dimeric structures are formed from the Nd(3+) ion through the Yb(3+) ion (9-coordinate), and a monomeric structure results for Lu(3+) (8-coordinate). The relaxivity studies presented here suggest that the high relaxivity of the Gd(tpaa) complex is mainly the consequence of a shorter bound water proton Gd(III) distance associated with a probable water coordination equilibrium between tris(aqua) and bis(aqua) complexes, giving raise to a mean number of coordinated water molecules q > 2. Both effects are strongly related to the ligand flexibility, which allows for a large volume available for water binding. The observed rapid water exchange rate is probably due to the presence of a low energy barrier between 10-, 9-, and 8- coordinate geometries. Although the low solubility of the Gd complex of tpaa prevents its practical application as an MRI contrast agent, the straightforward introduction of substituents on the pyridine rings allows us to envisage ligands with a higher water solubility, containing functional groups leading to macromolecular systems with very high relaxivity. PMID- 11735487 TI - 6-carboxamido-5,4-hydroxypyrimidinones: a new class of heterocyclic ligands and their evaluation as gadolinium chelating agents. AB - A previously unexplored class of heterocyclic bidentate chelating groups, 6 carboxamido-5,4-hydroxypyrimidinones (6-substituted-HOPYs), have been synthesized by two routes that provide a flexible entry into this ligand system. These are related to, but distinct from, the hydroxypyridonates and have been characterized in this study as a gadolinium chelating agent for magnetic resonance imaging (MRI) applications. The complex Gd[TrenHOPY] demonstrates high stability and high selectivity relative to other ions of biological interest, such as Zn(II) and Ca(II). These stability constants are comparable to those demonstrated by the previously studied 3,2-pyridinone analogues, however, the 5,4-pyrimidinones are at least an order of magnitude more soluble in water. The proton relaxation properties of Gd[TrenHOPY] in water were measured as a function of magnetic field, pH, and temperature. These results support the description of Gd[TrenHOPY] as a complex with two coordinated water molecules in fast exchange with bulk water. In addition, the influence of exogenous anions and blood serum proteins has been investigated. The favorable contrast agent properties emerging from these studies are discussed. PMID- 11735488 TI - New molecular arrays based on a tin(IV) porphyrin scaffold. AB - Two new porphyrin arrays-a hexamer and a nonamer-have been synthesized and characterized by elemental analysis as well as mass, (1)H NMR, and UV-vis spectroscopic methods. The scheme of construction of these arrays employs a synthetic protocol involving sequential "organic" and "inorganic" reactions conducted, respectively, at the peripheral meso-phenyl ring and the central tin(IV) ion of the porphyrin scaffold. The architecture of the hexamer is such that it is based on a covalently linked tin(IV) porphyrin dimer, with each of the two tin(IV) centers trans-axially ligated to two free-base porphyrins, while the higher homologue features a tin(IV) porphyrin trimer as the basal unit, with its central metalloid ions having two free-base porphyrins as axial ligands. This extended, "axial-bonding"-type architecture of the new arrays has been investigated by the (1)H NMR method, which reveals characteristic ring-current induced shifts and coupling patterns for the resonances due to protons of the axial free-base porphyrin subunits. The presence of any ring-ring (basal-basal, basal-axial, or axial-axial) interaction in these arrays is not obvious from their UV-vis and redox potential data, which are close to those of the corresponding constituent monomeric species. On the other hand, their singlet state activities are quite different from those of the precursor reference compounds as probed by steady-state fluorescence. The results of the detailed investigations carried out on these hybrid, "bichromophoric" arrays have been interpreted in terms of the occurrence of intraarray, interchromophore energy- and electron-transfer reactions. PMID- 11735489 TI - Synthesis of enantiopure oxorhenium(V) and arylimidorhenium(V) "3 + 2" Schiff base complexes. X-ray diffraction, cyclic voltammetry, UV-vis, and circular dichroism characterizations. AB - Two new oxorhenium(V) and two new arylimidorhenium(V) complexes of the Schiff base ligands 2-hydroxybenzaldehyde-((1R,2S)-1-amino-2-indanol)imine (1) (H(2)L(1)) and 3-(1-adamantyl)-2-hydroxy-5-methylbenzaldehyde-((1R,2S)-1-amino- 2 indanol)imine (2) (H(2)L(2)) have been prepared from the reaction of the precursor Re(O)(PPh(3))(2)Cl(3), Re(NC(6)H(5))(PPh(3))(2)Cl(3), or Re(NC(6)H(4)OCH(3))(PPh(3))(2)Cl(3) and the free ligands H(2)L(1,2). The complexes Re(O)(HL(1))(L(1)) (3), Re(O)(HL(2))(L(2)) (4), Re(NC(6)H(5))(HL(1))(L(1)) (5), and Re(NC(6)H(4)OCH(3))(HL(1))(L(1)) (6) have been isolated and fully characterized by IR, (1)H NMR, circular dichroism, LRMS FAB, and elemental analysis. All the complexes have a chiral center at rhenium. A single enantiomer is obtained in all cases. Suitable crystals of 3 and 5 were used in X-ray structural determinations. Crystal data: (3) C(32)H(27)N(2)O(5)Re.CH(2)Cl(2), orthorhombic, P2(1)2(1)2(1), a = 9.5599(16) A, b = 9.9579(16) A, c = 31.712(5) A, V = 3018.9(9) A(3), T = 100(2) K, Z = 4. (5) C(40)H(38)N(3)O(5)Re, monoclinic, P2(1), a = 9.286(3) A, b = 18.759(6) A, c = 9.957(3) A, beta = 102.817(6) degrees, V = 1691.3(10) A(3), T = 100(2) K, Z = 2. The major characteristic of these complexes is the presence of two coordination modes for the Schiff base ligands on rhenium, a tridentate ligand (noted L(1,2)) and another bidentate ligand (noted HL(1,2)). In the latter, the -OH group of the indanol is free and tilts away from the coordination sphere. X-ray structural analyses in conjunction with circular dichroism were used to assign the absolute configuration at rhenium (C). Cyclic voltammetry, UV-vis, and circular dichroism data are presented and discussed. The complexes were found to be highly stable and to resist reduction even when treated with organic phosphanes. PMID- 11735490 TI - Hydroxo-bridged Cubane-type tetrairon(II) clusters supported by sterically hindered carboxylate ligands. AB - A series of hydroxo-bridged cubane-type tetrairon(II) clusters, [Fe(4)(mu OH)(4)(mu-O(2)CAr(4)(-)t(BuPh))(2)(mu-OTf)(2)L(4)] (L = C(5)H(5)N (1), 4 (t)BuC(5)H(4)N (2), 3-FC(5)H(4)N (3)), were synthesized by using a sterically hindered carboxylate ligand, 2,6-di(4-tert-butylphenyl)benzoate (Ar(4)( )t(BuPh)CO(2)(-)). Three different bridging units that mediate weak antiferromagnetic coupling interactions between the metal centers support the unprecedented cubane-type [Fe(4)(mu-OH)(4)](4+) cores in 1-3. The solution structures of 1 and 3 probed by FT-IR and (19)F NMR spectroscopy are consistent with the solid-state geometry determined by X-ray crystallography. Zero-field Mossbauer spectra of 1-3 at 4.2 K are characteristic of high-spin iron(II) centers in nearly identical coordination environments. Compound 1 undergoes two irreversible oxidation processes at ca -10 and +880 mV (vs Fc/Fc(+)), the former approaching quasi-reversible behavior with increased scan rates and a narrow potential sweep range. Comparisons are made with analogous known [Fe(4)X(4)](n)()(+) (X = O, S) units, and the structural integrity of tetrairon fragments upon a change in oxidation state is discussed together with some possible biological implications. PMID- 11735491 TI - Reactivity and derivatization of five-coordinate, chelated aluminum. AB - The convenient five-coordinate starting materials, Salen((t)Bu)AlCl (Salen((t)Bu) = N,N'-alkylene (or arylene) bis (3,5-di-tert-butyl-2-hydroxybenzylideneamine) (1 4) can be used in a wide range of reactions to form five-coordinate aluminum compounds. Herein, these reagents were used to produce new five-coordinate azides, LAlN(3) (L = Salen((t)Bu) (5), Salpen((t)Bu) (6), and Salomphen((t)Bu) (7)) through trimethylsilylhalide elimination. The decomposition of the azides produce first hydroxide (LAlOH (L = Salen((t)Bu) (8)) and, subsequently in the presence of chlorotrimethylsilane, the siloxide compounds, LAlOSiMe(3) (L = Salen((t)Bu) (9), Salpen((t)Bu) (10), and Salomphen((t)Bu) (11)). Alkane elimination reactions may also be used to access this type of compound as evidenced by the formation of Salomphen((t)Bu)AlOSiPh(3) (12). Additionally, the first structurally characterized five-coordinate monomeric amide, Salcen((t)Bu)AlN(SiMe(3))(2) (13), can prepared by a salt elimination utilizing Salcen((t)Bu)AlCl (4). The compounds were characterized by spectroscopic methods ((1)H and (27)Al NMR, MS, and IR) and, in the case of 2 (Salpen((t)Bu)AlCl), 3 (Salomphen((t)Bu)AlCl) 9, 11, 12, and 13, by X-ray analysis. Several of the compounds were explored as potential catalysts for the living polymerization of propylene oxide. PMID- 11735492 TI - Chromium corroles in four oxidation States. AB - We have isolated and characterized chromium complexes of 5,10,15 tris(pentafluorophenyl)corrole [(tpfc)H(3)] (1) in four oxidation states: [(tpfc(*))CrO][SbCl(6)] (6); [(tpfc)CrO] (2); [(tpfc)CrO][Cp(2)Co] (4); and [(tpfc)Cr(py)(2)] (3). Complex 6 was prepared both by electrochemical and chemical oxidation of 2; its formulation as a Cr(V)O ligand-radical species is based on UV-visible absorption as well as EPR measurements. Cobaltocene reduction of 2 gave 4; it was identified as a diamagnetic d(2) Cr(IV)O complex from its sharp (1)H NMR spectrum. Reaction of 2 with triphenylphosphine yielded a chromium(III) corrole, [(tpfc)Cr(OPPh(3))(2)] (5). Owing to its air sensitivity, 5 could not be isolated in the absence of excess OPPh(3). The structure of the Cr(III) bis-pyridine complex (3) was determined by X-ray crystallography (Cr-N distances: 1.926-1.952 A, pyrrole; 2.109, 2.129 A, pyridine). PMID- 11735493 TI - A comparative study of third-order nonlinear optical properties of silver phenylacetylide and related compounds via ultrafast optical Kerr effect measurements. AB - A comparative study of the third-order nonlinear optical properties, via the newly developed heterodyned optical Kerr effect (OHD-OKE) measurements, of silver phenylacetylide and related compounds is reported. [AgC[triple bond]CC(6)H(5)](n) (1) was found to exhibit efficient third-order nonlinear optical susceptibility chi((3)) of 2.4 x 10(-14) esu, and second hyperpolarizability gamma of 9.07 x 10( 32) esu. These results are compared with those of two related silver phenylacetylide compounds, namely, a double salt, (silver phenylacetylide).(silver tert-butylthiolate) [AgC[triple bond]CC(6)H(5).AgS(t C(4)H(9))](n) complex (2), and a cluster, triphenylphosphine silver phenylacetylide tetramer, [(C(6)H(5))(3)PAgC[triple bond]CC(6)H(5)](4) (3), as well as that of the related organic polymer polyphenylacetylene (4). These four compounds represent different types of phenylacetylide derivatives: 1 is an organometallic polymer, 2 a polymeric double salt, 3 a discrete metal cluster, and 4 an organic polymer. It was found that the third-order optical nonlinear response was enhanced by the incorporation of silver d electrons into the delocalized conjugated organic pi system, and its magnitude is highly dependent upon the extent of the pi delocalization. Specifically, the relative magnitudes of chi((3)) and gamma follow the order silver phenylacetylide polymer (1) > (silver phenylacetylide).(silver tert-butylthiolate) double salt (2) > polyphenylacetylene polymer (4) > tetrameric (triphenylphosphine silver phenylacetylide)(4) cluster (3). The observed trend may be attributed to the decreasing length of pi conjugation. It is interesting to note that the incorporation of Ag(I) into the polymeric framework of polyphenylacetylene enhances the chi((3)) by 25-fold for the same degree of polymerization (n = 7). The signs of chi((3)) and gamma, which are related to the response mechanisms, were found to be solvent dependent. PMID- 11735494 TI - Bimolecular electron and energy transfer reactivity of exchange-coupled dinuclear iron(III) complexes. AB - Bimolecular quenching between photosensitizers and exchange-coupled transition metal complexes has been studied in an effort to experimentally establish a link between Heisenberg spin exchange and chemical reactivity. The acceptors are members of the oxo/hydroxo-biscarboxylato class of dinuclear Fe(III) compounds, where protonation of the oxo bridge provides a means for modulating the magnitude of spin exchange within the cluster. Photoexcitation of solutions containing Ru(II) polypyridyl sensitizers and the Fe(III) complexes results in quenching of emission from the (3)MLCT excited state of the Ru(II) chromophores; nanosecond time-resolved absorption measurements demonstrate that quenching occurs, in part, by electron transfer. Decoupling electron transfer driving force (DeltaG(0)(ET)) from changes in the magnitude of spin exchange was achieved by varying the bridging carboxylate to afford a series of complexes of the form [Fe(2)O(H)(O(2)CR)(2)(Tp)(2)](n)(+) (n = 0, 1, 2). Electrochemical measurements reveal a greater than 500 mV shift in cluster reduction potential across the series (i.e., R = CH(3) to CF(3)), whereas variable-temperature magnetic susceptibility measurements demonstrate a corresponding invariance in spin exchange between the metal centers (J(oxo) = -119 +/- 4 cm(-1) and J(hydroxo) = 18 +/- 2 cm(-1) for H = -2JS(1).S(2)). Structural analyses suggest that reorganization energies (lambda) associated with electron transfer should be identical for all molecules within a given series (i.e., oxo or hydroxo bridged); likewise Deltalambda between the series is expected to be small. A comparison of quenching rates for the two extended series firmly establishes that neither reorganization energy nor electron transfer driving force considerations can account for differences in reactivity between oxo-bridged (large spin exchange) and hydroxo-bridged (small spin exchange) quenchers. Upon consideration of energy transfer contributions, it is determined that reactivity differences between the oxo- and hydroxo-bridged quenchers must lie in the relative rates of Dexter energy transfer and/or electron transfer, with the origin of the latter linked to something other than DeltaG(0)(ET) or lambda. Finally, the extent to which spin exchange within the dinuclear Fe(III) quenchers can be identified as the key variable influencing these reactivity patterns is discussed. PMID- 11735495 TI - Exploring the photocatalytic properties and the long-lifetime chemosensor ability of Cl(2)[Ru(Bpy)(2)L] (L = 2,5,8,11,14-pentaaza[15])-2,2'-bipyridilophane). AB - In this work a new water-soluble long-lifetime chemosensor, containing a polyamine unit connected to a complexed Ru(II) metal center, is described. Its crystal structure has been characterized by X-ray analysis. The polyamine macrocyclic unit is capable of anchoring cationic or anionic substrates, according to its protonation state. Examples of electron transfer involving the ruthenium complex core and the bound substrate are presented. The photocatalytic ability of such a system is illustrated by the oxidation of iodide to iodine promoted by light absorption at 436 nm. PMID- 11735496 TI - Stacking efficiency of diselenadiazolyl pi-dimers. Consequences for electronic structure and transport properties. AB - The preparation and crystal structure of 5-cyanofuran-2-[1,2,3,5 diselenadiazolyl], [RCN(2)Se(2)] (R = 5-cyanofuran), is reported. Crystal data for C(6)H(2)ON(3)Se(2): monoclinic, space group P2(1), a = 7.1121(7), b= 20.541(2), c =20.923(2) A, beta = 99.785(1) degrees, Z = 16. The crystal structure consists of diselenadiazolyl pi-dimer stacks running parallel to the x direction; the asymmetric unit consists of four pi-dimer units. The dimers are aligned into snakelike ribbons along the y direction, with consecutive dimers linked by head-to-tail CN-Se contacts. Each pi-dimer stack is bordered by two out of-register stacks, but most interstack Se-Se contacts lie outside the van der Waals separation. Along the pi-dimer stacks, the intradimer Se-Se distances range from 3.183(10) to 3.294(1) A, and the interdimer Se-Se distances range from 3.826(1) to 3.945(1) A. Like other pi-dimer stacked diselenadiazolyls, [C(6)H(2)ON(3)Se(2)](2) is diamagnetic over the temperature range 4-380K. Variable temperature single-crystal conductivity measurements reveal a room temperature conductivity near 10(-5) S cm(-1) and provide a calculated band gap of 0.72 eV. The structural results and transport properties are interpreted in the light of Extended Huckel band structure calculations. PMID- 11735497 TI - Mixed chloride/phosphine complexes of the dirhenium core. 10. Redox reactions of an edge-sharing dirhenium(III) non-metal-metal-bonded complex, Re(2)(mu Cl)(2)Cl(4)(PMe(3))(4). AB - Reduction and oxidation reactions of the dirhenium(III) non-metal-metal-bonded edge-sharing complex, Re(2)(mu-Cl)(2)Cl(4)(PMe(3))(4) (1), have been studied. Several new mono- and dinuclear rhenium compounds have been isolated and structurally characterized in the course of this study. Reductions of 1 with 1 and 2 equiv of KC(8) result in an unusual face-sharing complex having an Re(2)(5+) core, Re(2)(mu-Cl)(3)Cl(2)(PMe(3))(4) (2), and a triply bonded Re(II) compound, 1,2,7,8-Re(2)Cl(4)(PMe(3))(4) (3), respectively. Two-electron reduction of 1 in the presence of tetrabutylammonium chloride affords a new triply bonded complex of the Re(2)(4+) core, [Bu(n)()(4)N][1,2,7-Re(2)Cl(5)(PMe(3))(3)] (4). Oxidation of 1 with NOBF(4) yields a Re(IV) mononuclear compound, trans ReCl(4)(PMe(3))(2) (5). Two isomers of the monomeric Re(III) anion, [ReCl(4)(PMe(3))(2)](-) (6, 7), have been isolated as side products. The crystal structures of compounds 2 and 4-7 have been determined by X-ray crystallography. The Re-Re distance in the face-sharing complex 2 of 2.686(1) A is relatively short. The metal-metal bond length in anion 4 of 2.2354(7) A is consistent with the usual values for the triply bonded Re(2)(4+) core compounds. In addition, a cis arrangement of trimethylphosphine ligands in the starting material 1 is retained upon reduction in the dinuclear products 2-4. PMID- 11735499 TI - Stereochemical influences on intervalence charge transfer in homodinuclear complexes of ruthenium. PMID- 11735498 TI - Two polymeric compounds built from mononuclear and tetrameric squarate-copper(II) complexes by deprotonation of 3,3-bis(2-imidazolyl)propionic acid (HBIP). Synthesis, crystal structure, and magnetic characterization of [Cu(HBIP)(BIP)](C(4)O(4))(1/2).2H(2)O and [[Cu(BIP)(OH(2))](4)(mu C(4)O(4))](ClO(4))(2).4H(2)O. AB - Two polynuclear copper(II)-squarate compounds of formulas [Cu(HBIP)(BIP)](C(4)O(4))(1/2).2H(2)O (1) and [[Cu(BIP)(OH(2))](4)(mu C(4)O(4))](ClO(4))(2).4H(2)O (2) (HBIP = 3,3-bis(2-imidazolyl)propionic acid) have been synthesized and characterized by single-crystal X-ray diffraction. Both compounds crystallize in the triclinic system, space group P1, with a =7.947(1) A, b =12.327(4) A, c = 13.150(3) A, alpha = 113.91(2) degrees, beta = 99.85(2) degrees, gamma = 90.02(2) degrees for compound 1 and a = 8.010(1) A, b = 13.073(1) A, c = 14.561(1) A, alpha = 72.13(1) degrees, beta = 80.14(1) degrees, gamma = 84.02(1) degrees for compound 2. The structure of compound 1 can be viewed as made up of [Cu(HBIP)(BIP)] units linked together by the BIP carboxylate groups to form a one-dimensional chain structure along the a axis in the crystal. The copper ion is five-coordinated (CuN(4)O chromophore) with BIP and HBIP acting as tridentate and bidentate ligands, respectively. The coordination geometry is intermediate between SP and TBP. The structure of compound 2 is made of infinite chains built from cationic tetranuclear [[Cu(BIP)(OH(2))](4)(mu-C(4)O(4))](2+) complex units, two uncoordinated perchlorate anions, and four water molecules of crystallization. The squarato group bridges the copper(II) ions, while BIP acts as a tridentate ligand, connecting through its carboxylate group the tetrameric units along the a axis. The two crystallographically independent copper(II) ions are pentacoordinated within a distorted square-based pyramid. Electronic and EPR spectra are consistent with the crystallographic data. Both compounds follow a Curie-Weiss law with very low values of theta (-0.13 and +0.12 K). In compound 2, the weak ferromagnetism interaction is discussed on the basis of the structural features and correlated with published magnetostructural data on similar squarato bridged copper(II) compounds. PMID- 11735500 TI - Structure and characterization of the first metal complex of dithiaporphyrin: Ru(S(2)TTP)Cl(2). PMID- 11735501 TI - Synthesis and characterization of diruthenium complexes in low oxidation states. Formation of mono- and bis-CO adducts. PMID- 11735502 TI - Isolation of a new nonaborane cluster form: arachno-B(9)H(11).(PPh(3))(2). PMID- 11735503 TI - A facile and environmentally friendly disposal of sodium and potassium with water. PMID- 11735504 TI - Total synthesis of (+)-ampullicin and (+)-isoampullicin: two fungal metabolites with growth regulatory activity isolated from Ampulliferina sp. 27. AB - The total synthesis of the growth regulators (+)-ampullicin 1 and (+) isoampullicin 2 from (R)-(-)-carvone 5 was accomplished by application of an 18 step sequence with 4.5% overall yield. The crucial step of the synthetic strategy lies on the internal displacement of tosylate 13 by means of the lactone enolate. In this way, access was opened to the tricyclic core present in these biologically active sesquiterpenic amides. A Horner-Emmons reaction between the carbaldehyde 16 and the phosphonate 22 led us to the stereoselective preparation of (+)-ampullicin 1. Standard transformation of 1 into the thermodynamically more stable geometric isomer (+)-isoampullicin 2 was trivial. The absolute configuration of both amides was established by X-ray analysis of a sample of synthetic (+)-isoampullicin 2. PMID- 11735505 TI - A convenient reductive deamination (hydrodeamination) of aromatic amines. AB - Reductive deamination (hydrodeamination) of aromatic amines can be conveniently carried out by amination of the corresponding arylamine methanesulfonamides using chloroamine under alkaline conditions. The intermediate aryl methanesulfonylhydrazines directly eliminate methanesulfinic acid, affording diazenes which extrude nitrogen affording the desired deaminated products. Both sulfonamide formation and reduction reactions occur in high yield and are compatible with a variety of functional groups. PMID- 11735506 TI - Nanostructures via noncovalent synthesis: 144 hydrogen bonds bring together 27 components. AB - This paper describes the spontaneous and reversible assembly of approximately 20 kDa synthetic hydrogen-bonded assemblies via the formation of 144 cooperative hydrogen bonds. These nanostructures ( approximately 3.0 x 5.5 nm), consisting of 27 different components, have been carefully characterized using a combination of (1)H NMR spectroscopy, MALDI-TOF MS using Ag(+)-labeling, gel permeation chromatography, and CD spectroscopy. PMID- 11735507 TI - Anion allosteric effect in the recognition of tetramethylammonium salts by calix[4]arene cone conformers. AB - Rigid calix[4]arene cone conformers, which are efficient receptors for quaternary ammonium salts, are usually obtained through the functionalization of their lower rim with suitable groups. Using flexible cone conformer of calix[4]arene, bearing four 4-hydroxybenzyl groups as cooperative and rigidifying structural elements at the upper rim of the calix, which act as anion binding groups, a new heteroditopic cavitand, 7, was synthesized. Whereas the tetramethoxy derivative 8 does not show any complexing ability, its tetrahydroxy analogue 7 recognizes tetramethylammonium salts with high efficiency. The binding abilities of this new receptor toward a series of tetramethylammonium salts (tosylate, chloride, acetate, trifluoroacetate, and picrate) have been investigated in CDCl(3) solution and compared to the monotopic and rigidified, through the lower rim, cone biscrown-3-calix[4]arene 9. The results obtained confirmed that in CDCl(3) ion pairing strongly affects binding. In particular, the rigid monotopic receptor 9 experiences good efficiency toward tetramethylammonium salts having anions with low ion-pairing ability such as trifluoroacetate or picrate. On the contrary, for the new heteroditopic cavitand 7, a reverse order of efficiency was found. In the latter case a different complexation mode was hypothesized in which the tetramethylammonium cation is deeply entrapped into the host cavity and its counteranion participates to the recognition process by coordination via hydrogen bonding by the four OH groups. To further support the role of the anion in the recognition process, a "dual host" approach, employing 7 or 9 in the presence of a specific receptor for chloride anion (10), was utilized. Molecular modeling studies confirmed that in the complexes formed by 7 and TMA salts the counteranion is involved in hydrogen bonding with the host OH groups and that the guests are bound as ligand-separated ion pairs. PMID- 11735508 TI - Synthesis of cyclodextrin-based carbohydrate clusters by photoaddition reactions. AB - The syntheses of homogeneous cyclodextrin-based carbohydrate clusters, persubstituted with beta-D-thioglucosyl or D-thiolactosyl residues on either (a) the primary face, (b) the secondary face, or (c) both the primary and the secondary faces of their cyclodextrin tori, are described. The key step in the synthetic methodology, namely the attachment of the carbohydrate residues to the cyclodextrin torus, proceeds in moderate-good yields (42-70%) by the photoaddition of thiol groups, positioned at the anomeric centers of the carbohydrate residues, to allyl ether functions on the cyclodextrins. Facile removal of protecting groups then affords the free cluster compounds. Extensive 1 D and 2-D NMR spectroscopic investigations were performed on these compounds to determine their structures and establish their homogeneities, and a brief computer molecular modeling study allowed estimates of the dimensions of the clusters to be determined. PMID- 11735509 TI - Design, synthesis, and evaluation of 9-D-ribityl-1,3,7-trihydro-2,6,8 purinetrione, a potent inhibitor of riboflavin synthase and lumazine synthase. AB - Reduction of 5-nitro-6-D-ribitylaminouracil (9) afforded 5-amino-6-D ribitylaminouracil (1), which reacted with ethyl chloroformate to yield 5 ethylcarbamoyl-6-D-ribitylaminouracil (12). The latter compound was cyclized to 9 D-ribityl-1,3,7-trihydropurine-2,6,8-trione (13), which was found to be a relatively potent inhibitor of both Escherichia coli riboflavin synthase (K(i) 0.61 microM) and Bacillus subtilis lumazine synthase (K(i) 46 microM). Molecular modeling of the lumazine synthase-inhibitor complex indicated the possibility for hydrogen bonding between the Lys135 epsilon-amino group of the enzyme and both the 8-keto group and the 4'-hydroxyl group of the ligand. A bisubstrate analogue of the riboflavin synthase-catalyzed reaction, 1,4-bis[1-(9-D-ribityl-1,3,7 trihydropurine-2,6,8-trionyl)]butane (18), was also synthesized using a similar route and was found to be inactive as an inhibitor of both riboflavin synthase and lumazine synthase. PMID- 11735510 TI - Synthesis and physical properties of protein core mimetics. AB - A series of mimetic cores composed of a synthetic scaffold and amino acids have been constructed and their properties investigated in chloroform. A relative measure of H-bond strength was obtained by comparing temperature coefficients derived from variable-temperature (1)H NMR experiments. Although most templates had a strong H-bond, only a single template composed of D- and L-phenylalanines was able to form two strong H-bonds. Templates containing D- and L-leucines formed only a single H-bond. The results of these studies suggest that aromatic edge-to-face interactions provide greater stabilization energy than aliphatic aromatic interactions in the tightly packed hydrophobic cores of proteins. Partial structures of the templates were derived by analyzing a series of two dimensional (1)H NMR spectra and performing molecular mechanics calculations using AMBER and MMFF94 force fields. PMID- 11735511 TI - Stereoselective synthesis of trifluoro- and monofluoro-analogues of frontalin and evaluation of their biological activity. AB - The stereoselective synthesis of both enantiomers of trifluoro frontalin (-) (1S,5R)- and (+)-(1R,5S)-8, as well as of diastereomeric monofluoro frontalines ( )-(1R,2R,5R)-18 and (-)-(1R,2S,5R)-20, analogues of the bioactive component of the aggregation pheromone of the Scolytidae insect family, has been accomplished starting from (-)-(1R)- and (+)-(1S)-menthyl (S)-toluene-4-sulfinate as a source of chirality and methyl trifluoroacetate or fluoroacetate, respectively, as sources of fluorine. The C-1 stereocenters were installed via stereoselective epoxidation of beta-sulfinyl ketones 2 and 13 with diazomethane. The bicyclic core was obtained by totally stereocontrolled and chemoselective tandem Wacker oxidation/intramolecular ketalization of the intermediate unsatured sulfinyl diols 5, 15, and 19. Axially fluorinated (-)-20 elicited a strong electroantennographic response in laboratory tests on females of Dendroctonus micans, whereas equatorially fluorinated (-)-18 and the trifluoroanalogue (-)-8 showed modest responses. Field trials using (-)-20 were not indicative owing to the locally scarce population of D. micans, but it showed some attractiveness for other Coleoptera families. PMID- 11735512 TI - Synthesis of 4- and 5-substituted 1-hydroxyimidazoles through directed lithiation and metal-halogen exchange. AB - Electrophiles were introduced regioselectively at the 5-position of 1 (benzyloxy)imidazole by lithiation at C-5 after protection of C-2 with a chloro or a trimethylsilyl group. Subsequent treatment with an electrophile afforded 5 substituted 1-(benzyloxy)-2-chloroimidazoles 8-13 and 5-substituted 1 (benzyloxy)imidazoles 3-5, the 2-(trimethylsilyl) group being lost during workup. Electrophiles were introduced regioselectively at the 4-position of 1 (benzyloxy)imidazole by bromine-lithium exchange of 4-bromo-2-chloro-1 (benzyloxy)imidazoles, protected at C-5 with chloro or trimethylsilyl groups, followed by reaction with an electrophile. The 5-(trimethylsilyl) group was removed via base-catalyzed desilylation. Chlorine at C-2 and O-benzyl groups were removed by palladium-catalyzed hydrogenolysis. PMID- 11735513 TI - Molecular structure of substituted phenylamine alpha-OMe- and alpha-OH-p benzoquinone derivatives. Synthesis and correlation of spectroscopic, electrochemical, and theoretical parameters. AB - Thirteen C(6) para-substituted anilinebenzoquinones derived from perezone (PZ) (2 (1,5-dimethyl-4-hexenyl)-3-hydroxy-5-methyl-1,4-benzoquinone) were prepared to analyze the effect of the substituents on quinone electronic properties. The effect of a hydrogen bond between the alpha-hydroxy and carbonyl C(4)-O(4) groups was determined in perezone derivatives by substituting electron-donor and electron-acceptor groups such as -OMe, -Me, -Br, and -CN and comparing the -OH (APZs) and -OMe (APZms) derivatives. Reduction potentials of these compounds were measured using cyclic voltammetry in anhydrous acetonitrile. The typical behavior of quinones, with or without alpha-phenolic protons, in an aprotic medium was not observed for APZs due to the presence of coupled, self-protonation reactions. The self-protonation process gives rise to an initial wave, corresponding to the irreversible reduction reaction of quinone (HQ) to hydroquinone (HQH(2)), and to a second electron transfer, attributed to the reversible reduction of perezonate (Q(-)) formed during the self-protonation process. This reaction is favored by the acidity of the alpha-OH located at the quinone ring. To control the coupled chemical reaction, we considered both methylation of the -OH group (APZms) and addition of a strong base, tetramethylammonium phenolate (Me(4)N(+)C(6)H(5)O(-)), to completely deprotonate the APZs. Methylation led to recovery of reversible, bi electronic behavior (Q/Q(*)(-) and Q(*)(-)/Q(2)(-)), indicating the nonacidic properties of the NH group. The addition of a strong base resulted in reduction of perezonate (Q(-)) obtained from the acid-base reaction of APZs with Me(4)N(+)C(6)H(5)O(-) to produce the dianion radical (Q(*)(2)(-)). Although the nitrogen atom interferes with direct conjugation between both rings by binding the quinone with the para-substituted ring, the UV-vis spectra of these compounds showed the existence of intramolecular electronic transfer from the respective aniline to the quinone moiety. (13)C NMR chemical shifts of the quinone atoms provided additional evidence for this electron transfer. These findings were also supported by linear variation in cathodic peak potentials (E(pc)) vs Hammett sigma(p) constants associated with the different electrochemical transformations: Q/Q(*)(-), Q(*)(-)/Q(2)(-) for APZms or HQ/HQH(2) and Q(-)/Q(*)(2)(-) for APZs. The electronic properties of model anilinebenzoquinones were determined at a B3LYP/6-31G(d,p) level of theory within the framework of the density functional theory. Our theoretical calculations predicted that all the compounds are floppy molecules with a low rotational C-N barrier, in which the degree of conjugation of the lone nitrogen pair with the quinone system depends on the magnitude of the electronic effect of the substituents of the aniline ring. Natural charges show that C(1) is more positive than C(4) although the LUMO orbital is located at C(4). Hence, if the natural charge distribution in the molecule controls the first electron addition, this should occur at carbon atom C(1). If the process is controlled by the LUMO orbitals, however, electron addition would first occur at C(4). For the APZms series susceptibility of the first reduction wave to the substitution effect (rho(pi) = 147 mV) is lower than that of the second reduction wave (rho(pi) = 156 mV). Thus, the first, one-electron transfer in the quinone system is controlled by the natural charge distribution of the molecule and therefore takes place at C(1). PMID- 11735514 TI - Highly enantioselective Rh-catalyzed hydrogenation based on phosphine-phosphite ligands derived from carbohydrates. AB - A new class of efficient catalysts was developed for the asymmetric hydrogenation of alpha,beta-unsaturated carboxylic acid derivatives by synthesizing a series of novel phosphine-phosphite ligands (4a-d) derived from readily available D-(+) xylose. Excellent enantioselectivities (> 99%) were achieved under very mild reaction conditions (1 bar H(2) and 20 degrees C). Varying the biphenyl substituents in the phosphite moiety greatly affected the enantioselectivity in the hydrogenation reactions. The results also indicate that the sense of enantioselectivity is mainly controlled by the configuration of the phosphite moiety. (31)P[(1)H] NMR and kinetic studies on intermediates of the catalytic cycle show that the [Rh(P(1)-P(2))(enamide)]BF(4) species is the resting state and that the rate dependence is first order in rhodium and hydrogen pressure and zero order in enamide concentration. PMID- 11735515 TI - Asymmetric synthesis of (+)-allo-quercitol and (+)-talo-quercitol via free radical cycloisomerization of an enantiomerically pure alkyne-tethered aldehyde derived from a carbohydrate. AB - We describe for the first time the free radical cyclization of enantiomerically pure alkyne-tethered aldehydes obtained from a carbohydrate (6, 7). The synthesis of compounds 6 and 7 obtained from a derivative of D-ribose is reported. These radical precursors have been submitted to cyclization with tributyltin hydride plus azobisisobutyronitrile to yield, after ring closure, two carbocycles, respectively. These carbocycles have been obtained as mixtures of E and Z vinyltin isomers, but with excellent diastereoselection at the new stereocenter formed during the ring closure. After protodestannylation, only one diastereomer was detected and isolated. The absolute configuration at the new stereocenter formed during the carbocyclization has been established by detailed (1)H NMR analysis. The specific transformation of 7-methoxymethoxy-2,2-dimethyl-4 methylene-5-tert-butyldimethylsilyloxy-(3aR,5S,7S,7aS) perhydrobenzo[d][1,3]dioxole into optically pure (+)-allo-quercitol and (+)-talo quercitol is described. From these results, we conclude that under an appropriate choice of radical precursors and conditions, the synthesis of highly functionalized cyclohexane derivatives of biological interest is now available. PMID- 11735516 TI - Enols of amides. The effect of fluorine substituents in the ester groups of dicarboalkoxyanilidomethanes on the enol/amide and E-enol/Z-enol ratios. A multinuclei NMR study. AB - Condensation of phenyl isocyanate substituted by 4-MeO, 4-Me, 4-H, 4-Br, and 2,4 (MeO)(2) with esters CH(2)(CO(2)R)CO(2)R', R = CH(2)CF(3), R' = CH(3), CH(2)CF(3), CH(CF(3))(2), or R = CH(3), R' = CH(CF(3))(2) gave 17 "amides" ArNHCOCH(CO(2)R)CO(2)R' containing three, six, or nine fluorines in the ester groups. X-ray crystallography of six of them revealed that compounds with > or =6 fluorine atoms exist in the solid state as the enols of amides ArNHC(OH)=C(CO(2)R)CO(2)R' whereas the ester with R = R' = CH(3) was shown previously to have the amide structure. In the solid enols, the OH is cis and hydrogen bonded to the better electron-donating (i.e., with fewer fluorine atoms) ester group. X-ray diffraction could not be obtained for compounds with only three fluorine atoms, i.e., R = CH(2)CF(3), R' = CH(3) but the (13)C CP-MAS spectra indicate that they have the amide structure in the solid state, whereas esters with six and nine fluorine atoms display spectra assigned to the enols. The solid enols show unsymmetrical hydrogen bonds and the expected features of push-pull alkenes, e.g., long C(alpha)=C(beta) bonds. The structure in solution depends on the number of fluorine atoms and the solvent, but only slightly on the substituents. The symmetrical systems (R = R' = CH(2)CF(3)) show signals for the amide and the enol, but all systems with R not equal R' displayed signals for the amide and for two enols, presumably the E- and Z-isomers. The [Enol I]/[Enol II] ratio is 1.6-2.9 when R = CH(2)CF(3), R' = CH(3), CH(CF(3))(2) and 4.5-5.3 when R = CH(3), R' = CH(CF(3))(2). The most abundant enol display a lower field delta(OH) and a higher field delta(NH) and assigned the E-structure with a stronger O-H.O=C(OR) hydrogen bond than in the Z-isomer. delta(OH) and delta(NH) values are nearly the same for all systems with the same cis CO(2)R group. The [Enols]/[Amide] ratio in various solvents follows the order CCl(4) > CDCl(3) > CD(3)CN > DMSO-d(6). The enols always predominate in CCl(4) and the amide is the exclusive isomer in DMSO-d(6) and the major one in CD(3)CN. In CDCl(3) the major tautomer depends on the number of fluorines. For example, in CDCl(3,) for Ar = Ph, the % enol (K(Enol)) is 35% (0.54) for R = CH(2)CF(3,) R' = CH(3), 87% (6.7) for R = R' = CH(2)CF(3), 79% (3.8) for R = CH(3), R' = CH(CF(3))(2) and 100% (> or =50) for R = CH(2)CF(3), R' = CH(CF(3))(2). (17)O and (15)N NMR spectra measured for nine of the enols are consistent with the suggested assignments. The data indicate the importance of electron withdrawal at C(beta), of intramolecular hydrogen bonding, and of low polarity solvents in stabilizing the enols. The enols of amides should no longer be regarded as esoteric species. PMID- 11735518 TI - A C-H...O=C hydrogen bond? Intramolecular hydrogen bonding in a novel semirubin. AB - (4Z)-8-(5-Carboxypentyl)-9-butyl-2,3-diethyl-dipyrrin-1-one (1), a new analogue of xanthobilirubic acid, (4Z)-8-(carboxyethyl)-2,7,9-dimethyl-3-ethyl-dipyrrin-1 one, was synthesized in four steps from the known 2,3-diethyl-dipyrrin-1-one. Whereas xanthobilirubic acid (which is a model for one-half of bilirubin, the yellow pigment of jaundice) and its homologues with hexanoic and longer acid chains at C-8 engage only in intermolecular hydrogen bonding, 1 is found to engage in intramolecular hydrogen bonding. In CDCl(3) solution, dipyrrinone 1 adopts an anti-Z conformation, and its hexanoic acid COOH is hydrogen-bonded to the lactam H-N-C=O and to the pyrrole C(7)-H but not to the pyrrole NH. The latter constitutes an example of a hydrogen bond of the type C-H...O=C, weak and detected typically in crystals. Dipyrrinone 1 is found by vapor pressure osmometry to be monomeric in CHCl(3), but its methyl ester (2) tends toward being dimeric, like that of methyl xanthobilirubinate, which is dimeric. PMID- 11735517 TI - Improved preparation and use of room-temperature ionic liquids in lipase catalyzed enantio- and regioselective acylations. AB - Polar organic solvents such as methanol or N-methylformamide inactivate lipases. Although ionic liquids such as 3-alkyl-1-methylimidazolium tetrafluoroborates have polarities similar to these polar organic solvents, they do not inactivate lipases. To get reliable lipase-catalyzed reactions in ionic liquids, we modified their preparation by adding a wash with aqueous sodium carbonate. Lipase catalyzed reactions that previously did not occur in untreated ionic liquids now occur at rates comparable to those in nonpolar organic solvents such as toluene. Acetylation of 1-phenylethanol catalyzed by lipase from Pseudomonas cepacia (PCL) was as fast and as enantioselective in ionic liquids as in toluene. Ionic liquids permit reactions in a more polar solvent than previously possible. Acetylation of glucose catalyzed by lipase B from Candida antarctica (CAL-B) was more regioselective in ionic liquids because glucose is up to one hundred times more soluble in ionic liquids. Acetylation of insoluble glucose in organic solvents yielded the more soluble 6-O-acetyl glucose, which underwent further acetylation to give 3,6-O-diacetyl glucose (2-3:1 mixture). However, acetylation of glucose in ionic liquids yielded only 6-O-acetyl glucose (>13:1 and up to >50:1). PMID- 11735519 TI - Synthesis of di- to hexasaccharide 1,2-linked beta-mannopyranan oligomers, a terminal S-linked tetrasaccharide congener and the corresponding BSA glycoconjugates. AB - Homo oligomers of (1-->2)-beta-D-mannopyranosyl residues have been synthesized in order to study the unique immunological properties of the cell wall mannan of C. albicans. p-Chlorobenzyl-protected ulosyl bromide (2) in combination with the sterically hindered, participating solvent, pivaloyl nitrile, facilitated a new approach for the synthesis of these unique homooligomers ranging from disaccharide up to hexasaccharide. The glycosyl donor 2 demonstrates high diastereoselectivity over both the glycosylation and subsequent reduction step and minimizes the number of protecting group manipulations necessary for the synthesis. Congeners of the (1-->2)-beta-D-mannotetraose were synthesized containing a terminal S-linked (1-->2)-beta-D-mannopyranosyl residue. Deprotection of these compounds afforded the propyl glycosides as well as oligomers with amino terminated aglyconic tethers. The tethers were generated from the oligosaccharide allyl glycosides by photoaddition with 2 aminoethanethiol. The functionalized haptens were coupled to BSA via squarate conjugation, and the degree of incorporation was established by TOF mass spectrometry. PMID- 11735520 TI - Preparation of (R)- and (S)-N-protected 3-hydroxypyrrolidines by hydroxylation with Sphingomonas sp. HXN-200, a highly active, regio- and stereoselective, and easy to handle biocatalyst. AB - Hydroxylation of N-benzylpyrrolidine 8 with resting cells of Sphingomonas sp. HXN 200 gave N-benzyl-3-hydroxypyrrolidine 15 in 53% ee (S) with an activity of 5.8 U/g CDW. By changing the "docking/protecting group" in pyrrolidines, hydroxylation activity and enantioselectivity were further improved and the enantiocomplementary formation of 3-hydroxypyrrolidines was achieved: hydroxylation of N-benzoyl-, N-benzyloxycarbonyl-, N-phenoxycarbonyl-, and N-tert butoxycarbonyl-pyrrolidines 9-12 gave the corresponding 3-hydroxypyrrolidines 16 19 in ee of 52% (R), 75% (R), 39% (S), and 23% (R), respectively, with an activity of 2.2, 16, 14, and 24 U/g CDW, respectively. Simple crystallizations increased the ee of 16-18 to 95% (R), 98% (R), and 96% (S), respectively. Hydroxylation of pyrrolidines 8-12 with soluble cell-free extracts of Sphingomonas sp. HXN-200 and equimolar NADH gave 3-hydroxypyrrolidines 15-19 in nearly the same ee as the products generated by whole cell transformation, suggesting that this strain possesses a novel soluble alkane monooxygenase. Cells of Sphingomonas sp. HXN-200 were produced in large amounts and could be stored at -80 degrees C for 2 years without significant loss of activity. The frozen cells can be thawed and resuspended for biohydroxylation, providing a highly active and easy to handle biocatalyst for the regio- and stereoselective hydroxylation of nonactivated carbon atoms. These cells were used to prepare 1.0-3.2 g (66.4-93.5% yield) of 3-hydroxypyrrolidines 16-19 by hydroxylation of pyrrolidines 9-12 on 0.9-2 L scale. Preparative hydroxylation was also achieved with growing cells as biocatalysts; hydroxylation of pyrrolidine 11 on 1 L scale gave 1.970 g (79.7% yield) of 3-hydroxypyrrolidine 18. PMID- 11735521 TI - The stereochemical outcome of electrophilic addition reactions on the 5,6-double bond in the spinosyns. AB - The electrophilic addition of reagents to the 5,6-double bond in spinosyn A and spinosyn D systems occurred with high pi-diastereofacial selectivity. Addition occurred preferentially from the beta face of the molecule with selectivities ranging from 5:1 to better than 30:1. Various NMR properties were investigated in order to distinguish the beta and alpha isomers with the help of theoretical models of the products. These NMR properties include a (13)C gamma effect to C-11 and vicinal coupling between H-4 and H-5. To help rationalize the selectivity, computational studies on the transition states for epoxidation were calculated using density functional theory. The results indicate that beta epoxidation is favored and that the geometries of the transition structures are consistent with torsional steering being the source of the selectivity. PMID- 11735522 TI - The reactions of diaminomaleonitrile with isocyanates and either aldehydes or ketones revisited. AB - A reinvestigation of the reactions of urea derivatives of diaminomaleonitrile 2 with aldehydes or ketones in the presence of triethylamine has established that the products of these reactions are not pyrimidino[5,4-d]pyrimidines 9 as previously reported, but 8-oxo-6-carboxamido-1,2-dihydropurines 12, which are oxidized rapidly in air to the corresponding 6-carboxamidopurines 13. Similarly, the reaction of Schiff base derivatives of DAMN 5 with isocyanates in the presence of triethylamine gives the substituted 2-oxoimidazoles 20 and not the pyrimidine derivatives 8 as previously claimed. The compounds 20 cyclize in solution and are easily oxidized to 8-oxopurine-6-carbonitriles 22, which give the same 8-oxopurine-6-carboxamides 13 upon further hydrolysis. PMID- 11735523 TI - Synthesis of bipyridylene-bridged bisporphyrin by nickel-mediated coupling reaction: ON-OFF control of cofacial porphyrin unit by reversible complexation. AB - Novel bipyridylene-bridged bisporphyrin 1a, in which two porphyrin units were attached directly to symmetrical 4,4'-positions of the 2,2'-bipyridyl group, was synthesized by a nickel(0)-mediated homocoupling reaction of 5,10,15-tris(n heptyl)-20-(2'-bromo-4'-pyridyl)porphyrinatozinc (3a) in 58% yield. Spatial geometries of two porphyrins in 1a were regulated by reversible complexation of the bipyridyl part with PdCl(2). Thus, the addition of 2.2 equiv of palladium chloride to 1a converted the freely rotating conformation to the cofacial bisporphyrin 2a. The subsequent addition of 4,4'-dimethyl-2,2'-bipyridine 9 regenerated the initial bisporphyrin 1a. PMID- 11735524 TI - Azomethine ylide cycloaddition/reductive heterocyclization approach to oxindole alkaloids: asymmetric synthesis of (-)-horsfiline. AB - The intermolecular [3 + 2] annulation of azomethine ylides with 2(2 nitrophenyl)acrylate dienophiles followed by reductive heterocyclization affords the spiro(indole-pyrrolidine) ring system. Hence, this enable us to accomplish a concise and highly enantioselective synthesis of (-)-horsfiline 1, based on chiral auxiliary-directed pi-face discrimination in the 1,3-dipolar cycloaddition of (1S,2R)-2-phenyl-1-cyclohexyl ester 4f with N-methylazomethine ylide. PMID- 11735525 TI - Solid-phase synthesis of oligourea peptidomimetics employing the Fmoc protection strategy. AB - A solid-phase-Fmoc-based-synthesis strategy is described for oligourea peptidomimetics as well as a convenient general synthesis approach for the preparation of the required building blocks 5a-j and 5k. These are suitable for use in peptide or robot synthesizers, which is illustrated by the synthesis of oligourea peptidomimetics of part of Leu-enkephalin (10) and a neurotensin derivative (17). PMID- 11735526 TI - Total synthesis of cystothiazoles A and C. AB - An efficient pathway culminating in the enantiocontrolled preparation of cystothiazoles A and C has been described. The cystothiazoles demonstrate potent antifungal activity and function as novel inhibitors of mitochondrial oxidation at a specific site on the cytochrome bc(1) complex. These studies outline a general and flexible plan that can be readily adapted for the synthesis of a variety of related five-membered heterocyclic systems and for biological investigations of structure-activity relationships. The core [2,4']bisthiazole component 8 was prepared in six steps, and the use of the Horner-Emmons olefination to yield the alpha,beta-unsaturated ester 10 set the stage for an asymmetric Evans aldol process, which established the required C(4)/C(5) stereochemistry. Finally the cystothiazoles A and C were prepared via a stereocontrolled O-alkylation of the precursor beta-keto esters 22a and 22b. PMID- 11735527 TI - On the mechanism of conversion of N-acyl-4-acyloxy-beta-lactams into 2 substituted 1,3-oxazin-6-ones. Can a low-barrier transition state be antiaromatic? AB - The mechanism of the conversion of N-acyl-4-acyloxy-beta-lactams into 1,3-oxazin 6-ones has been investigated using ab initio and density functional theories. It has been found that two pseudopericyclic reactions are involved in the whole process. The first key reaction is a retro-[4-exo-dig] cyclization instead of a thermal conrotatory electrocyclic ring opening. Magnetic characterization of the corresponding transition structure shows antiaromatic character, despite the low activation energy associated with this process. The second step is very exothermic and has no activation barrier. It corresponds to another pseudopericyclic reaction instead of a six-electron disrotatory electrocyclization. These results confirm that there is no correlation between aromaticity and pseudopericyclic reactions. In contrast, thermal-symmetry-allowed pericyclic reactions are always aromatic. Therefore, magnetic analysis of the corresponding transition structures constitutes a useful tool to distinguish between both kinds of processes. PMID- 11735528 TI - A straightforward stereoselective synthesis of D- and L-5-hydroxy-4-hydroxymethyl 2-cyclohexenylguanine. AB - A novel and facile synthesis of 5-hydroxy-4-hydroxymethyl-2-cyclohexenylguanine 1 is described. The key steps involve a Diels-Alder reaction of ethyl (2E)-3 acetyloxy-2-propenoate 2 as dienophile with Danishefsky's diene 3 to build up the six-membered ring skeleton, a Fraser-Reid reductive rearrangement of the adduct using LiAlH(4), and base-moiety introduction using a Mitsunobu reaction. Optically pure D- and L-1 were obtained via resolution of intermediate 7 with (R) (-)-methylmandelic acid. The synthetic procedure toward racemic 1 consists of only five steps and has proven to be highly efficient toward the synthesis of cyclohexenyl nucleosides. PMID- 11735529 TI - The Suzuki coupling reaction in the stereocontrolled synthesis of 9-cis-retinoic acid and its ring-demethylated analogues. AB - The thallium-accelerated Suzuki coupling reaction of tetraenyl iodide 19 and cyclohexenyl boronate 18 afforded ethyl 9-cis-retinoate (12) in high yield. Both coupling partners of the Suzuki reaction are better reacted immediately after generation from their precursors, tetraenylstannane 10 and cyclohexenyl iodide 13. The geometrically homogeneous tetraenylstannane 10, comprising the polyenic side chain of ethyl 9-cis-retinoate and its ring-demethylated analogues, was synthesized by a stereoselective Horner-Wadsworth-Emmons reaction. On the other hand, easily available cyclohexanones are ideal starting materials for preparation of the cyclohexenyl boronates required for the synthesis of the ring modified 9-cis-retinoic acid analogues. For hindered cyclohexanones, hydrazones were converted to cyclohexenyl iodides. Iodine-lithium exchange and trapping with B(OMe)(3) then afforded the cyclohexenyl boronates. If the precursor cyclohexanone has secondary carbons, the alkenyllithium species was conveniently formed by elimination of the C,N-dilithiated intermediate obtained upon treating the trisylhydrazone with n-BuLi (Shapiro reaction). None of the above procedures allowed the generation of the more substituted organolithium from 2 methylcyclohexanone. However, the alternative Stille cross-coupling of 34 and 10 afforded 9-cis-1,1-bisdemethylretinoic acid 7. Both Suzuki and Stille coupling reactions took place under mild conditions, and the preservation of the retinoid side-chain geometry was therefore secured. PMID- 11735530 TI - Fragmentation reactions of optically active trisubstituted cyclopropylcarbinyl radicals. AB - Fragmentation reactions of optically active trisubstituted cyclopropylcarbinyl radicals and their application to the synthesis of natural products are described. Preparation of the optically pure substrates for radical fragmentation reactions was efficiently accomplished by lipase-mediated desymmetrization of sigma-symmetrical 3-substituted-1,2-cyclopropanedimethanols. In the presence of a radical stabilizing group, e.g., aryl, ester, or alpha,beta-unsaturated ester, the fragmentation occurs selectively to generate the radical on the alpha-carbon of the group and provide the optically pure alkene derivatives. These derivatives possess three chemically distinct functionalities, making them excellent chiral building blocks for the construction of biologically active molecules. The synthetic usefulness of the procedure developed here has been demonstrated by an application to the enantioselective synthesis of both enantiomers of the key intermediate, 4-(3,4-methylenedioxybenzyl)dihydrofuran-2(3H)-one (54), for the total synthesis of biologically active lignans. PMID- 11735531 TI - A simplified and efficient route to 2'-O, 4'-C-methylene-linked bicyclic ribonucleosides (locked nucleic acid). AB - A novel efficient method for the synthesis of locked nucleic acid (LNA) monomers is described. The LNA 5',3'-diols containing thymine, 4-N-acetyl- and 4-N benzoylcytosine, 6-N-benzoyladenine, and 2-N-isobutyrylguanine as nucleobases were prepared via convergent syntheses. The method is based on the use of the common sugar intermediate 1,2-di-O-acetyl-3-O-benzyl-4-C-methanesulfonoxymethyl-5 O-methanesulfonyl-D-erythro-pentofuranose (8) that easily can be prepared from D glucose in multigram scale. Four different nucleobases were stereoselectively coupled to 8 using a modified Vorbruggen procedure to give the corresponding 4'-C branched nucleoside derivatives. Subsequent ring closing furnished the protected LNA nucleosides. The 5'-O-mesyl groups were efficiently displaced by nucleophilic substitution using sodium benzoate. Saponification of the 5'-benzoates followed by catalytic removal of the 3'-O-benzyl groups afforded the free LNA diols. The exocyclic amino groups of adenosine and cytidine were selectively acylated to give 4-N-acetyl- or 4-N-benzoyl-LNA-C and 6-N-benzoyl-LNA-A. The isobutyryl group of guanine was retained during the preparation of 2-N-isobutyryl-LNA-G. The LNA-T diol and base-protected LNA diols can be directly converted into LNA phosphoramidites for automated chemical synthesis of LNA containing oligonucleotides. PMID- 11735532 TI - A dirhodium(II)-carbenoid route to (-)- and (+)-Geissman-Waiss lactone: synthesis of (1R,7R,8R)-(-)-turneforcidine. AB - (-)- and (+)-Geissman-Waiss lactone, 4b, was efficiently prepared via the intramolecular C-H insertion reaction of the chiral nonracemic diazoacetates (-) 5a and (+)-5b catalyzed by dirhodium(II) tetrakis[methyl (5R and 5S)-3 phenylpropanoyl-2-imidazolidinone-5-carboxylate]. The cyclization was found to proceed with excellent regioselectivity and cis-diastereoselectivity. The bicyclic lactone (-)-4b was successfully used in the synthesis of the necine base, (-)-turneforcidine 2. PMID- 11735533 TI - Bridged bis(beta-cyclodextrin)s possessing coordinated metal center(s) and their inclusion complexation behavior with model substrates: enhanced molecular binding ability by multiple recognition. AB - To investigate quantitatively the cooperative binding ability of several beta cyclodextrin oligomers bearing single or multiligated metal center(s), the inclusion complexation behavior of four bis(beta-cyclodextrin)s (2-5) linked by 2,2'-bipyridine-4,4'-dicarboxy tethers and their copper(II) complexes (6-9) with representative dye guests, i.e., methyl orange (MO), acridine red (AR), rhodamine B (RhB), ammonium 8-anilino-1-naphthalenesulfonic acid (ANS), and sodium 6-(p toludino)-2-naphthalenesulfonate (TNS), have been examined in aqueous solution at 25 degrees C by means of UV-vis, circular dichroism, fluorescence, and 2D NMR spectroscopy. The results obtained indicate that bis(beta-cyclodextrin)s 2-5 can associate with one or three copper(II) ion(s) producing 2:1 or 2:3 bis(beta cyclodextrin)-copper(II) complexes. These metal-ligated oligo(beta-cyclodextrin)s can bind two model substrates to form intramolecular 2:2 host-guest inclusion complexes and thus significantly enhance the original binding abilities of parent beta-cyclodextrin and bis(beta-cyclodextrin) toward model substrates through the cooperative binding of two guest molecules by four tethered cyclodextrin moieties, as well as the additional binding effect supplied by ligated metal center(s). Host 6 showed the highest enhancement of the stability constant, up to 38.3 times for ANS as compared with parent beta-cyclodextrin. The molecular binding mode and stability constant of substrates by bridged bis- and oligo(beta cyclodextrin)s 2-9 are discussed from the viewpoint of the size/shape-fit interaction and molecular multiple recognition between host and guest. PMID- 11735534 TI - 1,4- and 1,3-dipolar reactivity of alpha-alkoxycarbonylcycloimmonium N-aminides with dipolarophiles: synthesis of new imidazo[2,1-f][1,2,4]triazinium inner salts. AB - 2-Alkoxycarbonylazolium N-aminides are interesting species, as they have the potential to act as efficient 1,4-dipole equivalents when they react with heterocumulenes, such as iso(thio)cyanates and carbodiimides. These reactions give heterobetaines containing the imidazo[2,1-f][1,2,4]triazinium system, in a formal [4 + 2] cyclocondensation process. These N-aminides, however, can also behave as 1,3-dipoles when they react with isocyanates to afford a cycloadduct that, depending on the position of alkoxycarbonyl group, can undergo a reversion process or a ring expansion to the more stable heterobetaine system. PMID- 11735535 TI - The tridurylsilylium and tridurylstannylium cations: free and not so free. AB - Allyltridurylsilane has been prepared and converted to the tridurylsilylium cation by treatment with electrophiles. Allyltridurylstannane has been prepared and converted to the tridurylstannylium cation under similar conditions. The (29)Si chemical shift of the silylium cation indicates that it is free and tricoordinate, whereas the (119)Sn chemical shift of the stannylium cation indicates that it maintains loose fourth coordination with the solvent or the anion. PMID- 11735536 TI - Solid-phase synthesis of a branched hexasaccharide using a highly efficient synthetic strategy. AB - The solid-phase synthesis of branched lacto-N-neohexaose derivative 1 occurring in human milk is described. The new building block of lactose 3 bearing the orthogonal temporary hydroxy protecting groups 9-fluorenylmethyloxycarbonyl (Fmoc) and levulinoyl (Lev) has been prepared. Its use, together with that of lactosamine donor 4, glucosamine donor 5, and O-galactosyl trichloroacetimidate 6, has enabled the preparation of hexasaccharide 22 following two different approaches in excellent overall yield (43%, 90% per step over eight steps). An additional key feature of this work is the successful use of newly prepared ester type linker 2, having a benzylic spacer connected to the anomeric oxygen. This linker presents the advantage of producing a benzylic anomeric moiety after cleavage from the polymer support, which could be easily removed to obtain the unprotected oligosaccharide 1. PMID- 11735537 TI - Structure-reactivity relationships in the pyridinolysis of N-methyl-N arylcarbamoyl chlorides in dimethyl sulfoxide. AB - Nucleophilic substitution reactions of N-methyl-N-arylcarbamoyl chlorides (YC(6)H(4)N(CH(3))COCl) with pyridines (XC(5)H(4)N) have been investigated in dimethyl sulfoxide at 45.0 degrees C. A striking trend in the selectivity parameters is that they are constant within experimental errors, rho(X) = -2.25 +/- 0.03, beta(X) = 0.42 +/- 0.01, and rho(Y) = 1.10 +/- 0.06, with changing reactivities of the electrophiles (deltasigma(Y)) and nucleophiles (deltasigma(X)), respectively, and this leads to a vanishingly small cross interaction constant, rho(XY) approximately equals beta(XY) approximately equals 0. The rate data can be expressed in the Ritchie N(+) type equation. Based on this and other results, the mechanism of nucleophile (pyridine) addition to the resonance- stabilized carbocation is proposed. It has been shown from the definition of beta(XY) (and rho(XY)) together with the Marcus equation that the high intrinsic barrier, DeltaG(0), in the intrinsic-barrier controlled reaction series is a prerequisite for such reactions in which the cross-interaction vanishes and the N(+) relationship holds. PMID- 11735538 TI - Photochemical reactions of substituted cyclopropenium salts. AB - Photochemical reactions of triphenylcyclopropenium tetrakis(pentafluorophenyl)gallate (TPCPGa) and diphenyl-(2-methoxy-1-naphthyl) cyclopropenium tetrakis(pentafluorophenyl)gallate (DPMNCPGa) (Chart 1) have been investigated in acetonitrile. Traces of water were required for the photochemical reactions to proceed. The disappearance of both TPCPGa and DPMNCPGa obeys zero order kinetics with rate constants (k) having a linear dependence on the concentration of water. Electron-transfer from water to the cyclopropenium cation is proposed as the primary process in the formation of the cyclopropenyl radical. The latter dimerizes leading to the photoproducts. PMID- 11735539 TI - Theoretical study of stability, structures, and aromaticity of multiply N confused porphyrins. AB - The total electronic energy and nucleus-independent chemical shift (NICS) of 95 isomers of N-confused porphyrin (NCP: normal porphyrin (N(0)CP), singly N confused porphyrin (N(1)CP), doubly N-confused porphyrin (N(2)CP), triply N confused porphyrin (N(3)CP), and fully N-confused porphyrin (N(4)CP)) have been calculated by the density functional theory (DFT) method. The stability of NCP decreased by increasing the number of confused pyrrole rings. Namely, the relative energies of the most stable isomers in each confusion level increased in a stepwise manner approximately by +18 kcal/mol: 0 (N(0)CP1), +17.147 (N(1)CP2), +37.461 (N(2)CPb3), +54.031 (N(3)CPd6), and +65.636 kcal/mol (N(4)CPc8). In this order, the mean plane deviation of these isomers increased from 0.000 to 0.123, 0.170, 0.215, and 0.251 A, respectively. The unusual tautomeric forms of pyrrole ring with an sp(3)-carbon were found in the stable forms of N(3)CP and N(4)CP. The NICS values at the mean position of the 24 core atoms were nearly the same for the most aromatic isomers regardless of the confusion level: -15.1280 (N(0)CP1), -13.8493 (N(1)CP2), -13.7267 (N(2)CPd1), -11.7723 (N(3)CPb5), and 13.6224 ppm (N(4)CPa6). The positive correlation between aromaticity and stability was inferred from the plots of NICS and the relative energy of NCP for N(0)CP, N(1)CP, and trans-N(2)CP. On the other hand, the correlation was negative for cis-N(2)CP, N(3)CP, and N(4)CP isomers. PMID- 11735541 TI - Alkynylcyclohexanol chairs and twist-boats: Co(2)(CO)(6) as a conformational switch. AB - Treatment of 1-[axial]-(trimethylsilylethynyl)cyclohexan-1-ol with dicobalt octacarbonyl results in a conformational ring flip such that the bulky dicobalt alkyne cluster moiety now occupies the favored equatorial site. However, when a 4 tert-butyl substituent is present, the coordinated alkynyl group retains its original axial or equatorial position. Complexation of trans-[diaxial]-1,4 bis(triphenylsilylethynyl)cyclohexane-1,4-diol brings about a chair-to-chair conformational inversion such that both cluster fragments now occupy equatorial sites. In contrast, cis-1,4-bis(triphenylsilylethynyl)cyclohexane-1,4-diol reacts with Co(2)(CO)(8) to yield the twist-boat conformer in which the two axial hydroxy substituents exhibit intra-molecular hydrogen bonding. Likewise, the corresponding reaction of cis-1,4-bis(trimethylsilylethynyl)cyclohexane-1,4-diol with Co(2)(CO)(8) leads to a twist-boat, but in this case, the molecules are linked through inter-molecular hydrogen bonds. Eight of these cobalt clusters have been characterized by X-ray crystallography, and the potential use of twist boats in synthesis is discussed. PMID- 11735540 TI - Stereoselective synthesis of delta-lactones from 5-oxoalkanals via one-pot sequential acetalization, Tishchenko reaction, and lactonization by cooperative catalysis of samarium Ion and mercaptan. AB - By the synergistic catalysis of samarium ion and mercaptan, a series of 5 oxoalkanals was converted to (substituted) delta-lactones in efficient and stereoselective manners. This one-pot procedure comprises a sequence of acetalization, Tishchenko reaction and lactonization. The deliberative use of mercaptan, by comparison with alcohol, is advantageous to facilitate the catalytic cycle. The reaction mechanism and stereochemistry are proposed and supported by some experimental evidence. Such samarium ion/mercaptan cocatalyzed reactions show the feature of remote control, which is applicable to the asymmetric synthesis of optically active delta-lactones. This study also demonstrates the synthesis of two insect pheromones, (2S,5R)-2-methylhexanolide and (R)-hexadecanolide, as examples of a new protocol for asymmetric reduction of long-chain aliphatic ketones. PMID- 11735542 TI - 6-bromopurine nucleosides as reagents for nucleoside analogue synthesis. AB - Surprisingly facile direct substitution reactions with acetyl-protected 6 bromopurine nucleosides are described. Included in the series of bromonucleosides studied is the guanosine derivative N(2)-2',3',5'-tetraacetyl-6-bromopurine ribonucleoside, the synthesis of which is reported here for the first time. Brominated nucleosides had not previously been considered optimal substrates for S(N)Ar reactions given the general reactivity trend for halogenated aromatic systems (i.e. F > Cl > Br > I). However, even weakly nucleophilic aromatic amines give high yields of the substitution products in polar solvents with these 6 bromopurine nucleosides. For primary aromatic amines, secondary aliphatic amines, and imidazole, reaction takes place only at C6, with no effect on the acetyl protected ribose. In addition, we report the first synthesis of 3',5'-di-O-acetyl 6-bromopurine-2'-deoxyribonucleoside and its reaction with an arylamine in MeOH in the absence of added metal catalyst. Thus, C6-arylamine derivatives of both adenosine and 2'-deoxyadenosine can be prepared via simple S(N)Ar reactions with the corresponding 6-bromo precursor. We also describe high yielding and C6 selective substitution reactions with 6-bromonucleosides using alcohol and thiol nucleophiles in the presence of added base (DBU). Finally, C6-bromonucleosides are shown to be readily hydrogenated to give purine or 2-aminopurine products in good yield. This work increases the arsenal of reactions and strategies available for the synthesis of nucleoside analogues as potential biochemical tools or new therapeutics. PMID- 11735543 TI - Preparation of vinylogous 2-sulfonylindolines by the palladium-catalyzed heteroannulation of o-iodoanilines with dienyl sulfones and their further transformation to indoles and carbazoles. AB - The palladium-catalyzed heteroannulation of o-iodoanilines with dienyl sulfones provides a convenient route to vinylogous 2-sulfonylindolines 3. The reaction proceeds in DMF/water in the presence of potassium carbonate and catalytic palladium(II) acetate and is compatible with both electron-donating and withdrawing substituents in the para position of the aniline, and with an alkyl substituent at C-2 of the dienyl sulfone. The indolines underwent oxidation with DDQ to afford the corresponding indoles 4. The latter were then employed as dienes in Diels-Alder reactions with dimethyl acetylenedicarboxylate (DMAD), methyl propiolate, or methyl acrylate. In the case of the latter two dienophiles, the cycloadditions were highly regioselective, affording the corresponding 1,3 products (with respect to the relative positions of the sulfone and ester groups), exclusively. The cycloadducts from acetylenic dienophiles were converted to the corresponding carbazoles by elimination of the sulfone moiety with DBU, and that from methyl acrylate was subjected to reductive desulfonylation and oxidation to the corresponding carbazole with DDQ. The method thus provides access to carbazoles with various substituents at the 3-, 4-, and 6-positions. PMID- 11735544 TI - Acid-catalyzed cyclization of 2,3-dibenzylidenesuccinates: synthesis of lignans (+/-)-cagayanin and (+/-)-galbulin. AB - Acid-catalyzed cyclizations of E,E-dibenzylidenesuccinate esters have been developed as an efficient synthetic route to 1-aryl-1,2-dihydronaphthalenes. This reaction has been used in the synthesis of the naturally occurring lignans (+/-) cagayanin and (+/-)-galbulin. PMID- 11735545 TI - Synthesis of carbazoles from N-(N,N-diarylamino)phthalimides with aluminum chloride via diarylnitrenium ions. AB - Generation of diarylnitrenium ions from N-(N,N-diarylamino)phthalimides by treatment with AlCl(3) in benzene or in 1,2-dichloroethane leads to formation of carbazoles by intramolecular C-C bond formation. The reaction proceeds in synthetically useful yields. PMID- 11735546 TI - Friedel-Crafts sulfonylation in 1-butyl-3-methylimidazolium chloroaluminate ionic liquids. AB - 1-Butyl-3-methylimidazolium chloroaluminate ionic liquids have been employed as an unconventional reaction media and as Lewis acid catalyst for Friedel-Crafts sulfonylation reaction of benzene and substituted benzenes with 4-methyl benzenesulfonyl chloride. The substrates exhibited enhanced reactivity, furnishing almost quantitative yields of diaryl sulfones, under ambient conditions. Studies concerning the effect of Lewis acidity of the ionic liquid on the initial extent of conversion of this reaction has been carried out. (27)Al NMR spectroscopy has been exploited as a tool to investigate the mechanistic details of the reaction. (27)Al NMR spectral studies show the predominance of [Al(2)Cl(7)](-) species in [bmim]Cl-AlCl(3), N = 0.67, acidic ionic liquid in the presence of 4-methyl benzenesulfonyl chloride, and after the reaction with the aromatic hydrocarbon, [AlCl(4)](-) species predominates. This change in speciation of aluminum can be attributed to the interaction of the Lewis acidic species [Al(2)Cl(7)](-) of the ionic liquid with the formed HCl during the sulfonylation reaction, which is evidenced by the control experiment. Preliminary investigations on Friedel-Crafts acylation further substantiate the argument. PMID- 11735547 TI - Bimolecular photoreduction of aromatic sulfoxides. AB - Photolysis of aromatic sulfoxides in the presence of alkoxides in alcoholic solvents provides a photochemical route to the corresponding sulfides. Other electron donors also give sulfide with various degrees of success. The reaction could also be carried out using carbazoles as sensitizers, and quantitative yields could be obtained using N-methylcarbazole in methanol. Evidence points toward a hydroxysulfuranyl radical as the key intermediate, and solvent effects point to heterolysis, rather than homolysis, as the step that breaks the S-O bond. PMID- 11735548 TI - Constraining of small-ring cyclic ether triads by stereodefined spiroannulation to an inositol orthoformate platform. Solution- and gas-phase alkali metal binding affinities for three- to five-membered ring structural combinations. AB - The structural features most conducive to complexation of the alkali metal ions Li(+), Na(+), and K(+) in a series of constrained inositol orthoformate derivatives have been probed in solution, in the solid state, and in the gas phase by electrospray ionization mass spectrometry. The eight spirotricyclic polyethers differ in the size of the rings containing the potentially ligating oxygen atoms. Although the ring sizes have been limited to three to five atoms inclusively, the combinations of oxirane, oxetane, and tetrahydrofuran are rather extensive and consist of many options. The overall trend for lithium ion affinity is [5.5.5] > [ 5.5.4] > [4.4.4] > [5.5.3] > [5.4.3] > [4.4.3] > [1.1.1] > [3.3.1], an ordering that correlates with the differing polarizabilities of the oxygen atoms, ease of alignment of the nonbonded electron pairs, and the overall size of the ligand as gauged by nonbonded O......O distances. PMID- 11735549 TI - A cobalt-phosphine complex directed Reformatsky approach to a stereospecific synthesis of the dolastatin 10 unit dolaproine (Dap). PMID- 11735550 TI - A general synthesis of 5'-azido-5'-deoxy-2',3'-O-isopropylidene nucleosides. PMID- 11735551 TI - The catalytic Sakurai reaction. PMID- 11735552 TI - Selective aerobic oxidation of alcohols with a combination of a polyoxometalate and nitroxyl radical as catalysts. PMID- 11735553 TI - Convergent synthesis of 3-arylated 1-hydroxypyrazoles via 3-metalated pyrazole-1 oxides. PMID- 11735554 TI - Synthesis of aziridine-2,2-dicarboxylates via 1,4-addition of N,O (bistrimethylsilyl)hydroxylamine to alpha,beta-unsaturated malonates. PMID- 11735555 TI - Enantiopure preparation of the two enantiomers of the pseudo-C(2)-symmetric N,N dibenzyl-1,2:4,5-diepoxypentan-3-amine. PMID- 11735556 TI - Practical synthesis of 1,3-O-di-tert-butylsilylene-protected D- and L-erythritols as a four-carbon chiral building block. PMID- 11735557 TI - On the mechanism of quinone formation from the Bergman cyclization: some theoretical insights. PMID- 11735558 TI - A novel approach for solid-phase synthesis of substituted imidazolines and bis imidazolines. PMID- 11735559 TI - Highly active, air-stable versatile palladium catalysts for the C-C, C-N, and C-S bond formations via cross-coupling reactions of aryl chlorides. PMID- 11735560 TI - High enantioselectivity and broad substrate specificity of a carbonyl reductase: toward a versatile biocatalyst. PMID- 11735561 TI - Bis(5-alkyl-2-furyl)(2-carboxyphenyl)methanes for the synthesis of tetracyclic isochromone derivatives. PMID- 11735563 TI - Conjugate hydrostannation of unsaturated esters by iodotin hydride ate complex. PMID- 11735562 TI - A nonisocyanate route to monodisperse branched polyurethanes. PMID- 11735565 TI - Ketone-DNA: a versatile postsynthetic DNA decoration platform. AB - [reaction: see text] A general strategy for the functional diversification of DNA oligonucleotides under physiological conditions was developed. We describe the synthesis of DNA molecules bearing ketone ports (ketone-DNA) and the efficient postsynthetic decoration of ketone-DNA with structurally diverse aminooxy compounds. PMID- 11735566 TI - 2-Aryl-2,2-difluoroacetamide FKBP12 ligands: synthesis and X-ray structural studies. AB - [structure: see text] 2-Aryl-2,2-difluoroacetamido-proline and pipecolate esters are high affinity FKBP12 ligands whose rotamase inhibitory activity is comparable to that seen for the corresponding ketoamides. X-ray structural studies suggest that the fluorine atoms participate in discrete interactions with the Phe36 phenyl ring and the Tyr26 hydroxyl group, with the latter resembling a moderate to-weak hydrogen bond. PMID- 11735567 TI - Diastereoselective addition reactions of furyl aldehydes using chiral boronates as auxiliary: application to the enantioselective synthesis of 2,3-disubstituted furyl alcohols. AB - [reaction: see text] The addition reactions of various nucleophiles to a furyl aldehyde bearing a chiral boronate at the C-3 position furnished chromatographically separable diastereomers. The R diastereoselection was more favorable when no additive was added. Surprisingly, when lithium alkoxides were selected as additives, the S diastereoselection is superior instead. Further transformation of C-B bonds to C-C bonds was achieved by using standard Suzuki coupling conditions to give optically active 2,3-disubstituted furyl alcohols. PMID- 11735568 TI - Efficient strategy for the synthesis of stereopentad subunits of scytophycin, rifamycin S, and discodermolide. AB - [reaction: see text] An efficient, simple method has been developed for the stereocontrolled synthesis of polypropionate stereopentads in high enantio- and diastereomeric purities. PMID- 11735569 TI - Cooperative and selective lithium complexation of 2,11,13,22-tetraaza-5,8,16,19- tetraoxa-1,12-dioxocyclodocosanes. AB - [reaction: see text] Ureyleno crown ethers 2 and 3 bind with 2 equiv of Li(+) cooperatively and selectively over other alkali metal ions such as Na(+), K(+), and Cs(+). The binding constant for 3 was found to be 3.0 x 10(7) (L/mol)(2). PMID- 11735570 TI - Intramolecular palladium(II)-mediated alkoxy carbonylation as a route to functionalized tetrahydropyrans. Synthesis of the C9-C32 segment of phorboxazole A. AB - [reaction: see text] Hydroxy alkene 12, synthesized stereoselectively from 2 methyloxazole-4-carboxaldehyde, underwent intramolecular methoxy carbonylation in the presence of palladium(II) acetate to give 13 in which all five stereogenic centers around the tetrahydropyran correspond to those in ring C of phorboxazole A. Aldehyde 15, derived from 13, was linked to hydroxy alkene 23 via a Wittig coupling, and the composite 25 was subjected to a second palladium(II) acetate mediated methoxy carbonylation to yield 26, accompanied by acetoxy ester 27. PMID- 11735571 TI - Unusual reagent control of diastereoselectivity in the 1,2-addition of hard carbon nucleophiles to C(6)-heteroatom substituted cyclohexenones. AB - [reaction: see text] A surprising and synthetically useful counterion-dependent reversal of diastereoselectivity was found in 1,2-additions of hard carbon nucleophiles to C(6)-heterosubstituted cyclohexenones. In general, Grignard reagents added syn to the C(6)-substituent and Li reagents added anti, although some exceptions were found. Selectivities could be increased in some cases by appropriate choice of solvent and/or cosolvent. PMID- 11735572 TI - Enantioselective synthesis for the antipodes of slagenins B and C: establishment of absolute stereochemistry. AB - [reaction: see text] The total synthesis for the (-)-antipode of slagenin B (12) and the (+)-antipode of slagenin C (13) was achieved with the condensation of glyoxal hydrate 10 and urea as the key step. The absolute stereochemistries of naturally isolated slagenins B and C were assigned to be (9R,11R,15R)-1b and (9R,11S,15S)-1c, respectively. PMID- 11735573 TI - Enantiodivergent synthesis of either enantiomer of ABCDE-ring analogue of antitumor antibiotic fredericamycin A via intramolecular [4 + 2] cycloaddition approach. AB - [reaction: see text] An intramolecular enantiodivergent synthesis of both enantiomers of the ABCDE-ring analogue 22 of fredericamycin A is reported. Key steps involve an intramolecular [4 + 2] cycloaddition of 17 and an aromatic Pummerer-type reaction of 19. A lipase-catalyzed enantioselective desymmetrization of prochiral diol 2 using 1-ethoxyvinyl 2-furoate 3 led to the pivotal intermediate (R)-4. PMID- 11735574 TI - Trehalose-based octopus glycosides for the synthesis of carbohydrate-centered PAMAM dendrimers and thiourea-bridged glycoclusters. AB - [structure: see text] The nonreducing disaccharide trehalose was modified into an octa-amino-functionalized core molecule to serve in the synthesis of carbohydrate centered PAMAM glycodendrimers and thiourea-bridged glycoclusters. PMID- 11735575 TI - General approach for the synthesis of sarpagine/macroline indole alkaloids. Enantiospecific total synthesis of the indole alkaloid trinervine. AB - [structure: see text] The total synthesis of the indole alkaloid trinervine 1 was accomplished in enantiospecific fashion in an overall yield of 20% (from the tetracyclic ketone 8) in 10 reaction vessels (12.5% from tryptophan methyl ester). The synthesis of the N(a)-H substituted macroline equivalent 2 was also completed in high yield via the same intermediate 13. The unique protection/hydroboration process developed here should provide a method to functionalize the C(19)-C(20) double bond in similar systems. PMID- 11735576 TI - Short total syntheses of both the putative and actual structures of the clerodane diterpenoid (+/-)-sacacarin by double annulation. AB - [reaction: see text] The putative structure of the naturally occurring clerodane diterpenoid (+/-)-sacacarin has been prepared in only 10 steps, six of which are C-C bond-forming steps, in a chemo-, regio-, and diastereoselective manner. The key part of the synthesis is the double annulation (double Michael, Pinner, and Dieckmann reaction) of a tethered carbon diacid and 3-butyn-2-one. A corrected structure for sacacarin is proposed, and the structure is proven by synthesis. PMID- 11735577 TI - Synthesis and ring-expanding oligomerization of an extremely strained macrocyclic aromatic ether-sulfone. AB - [reaction: see text] An extremely strained, 5-ring, macrocyclic aromatic ether sulfone, [-1,4-C(6)H(4)SO(2)-1,4-C(6)H(4)O-1,3-C(6)H(4)O-1,4-C(6)H(4)SO(2)-1,4 C(6)H(4)-], synthesized by nickel-catalyzed intramolecular coupling of a bis chlorophenylene-terminated precursor, undergoes fluoride-promoted ring expansion in dilute solution, giving a series of higher macrocyclic oligomers including cyclic dimer, trimer, and tetramer. The latter contains 20 aromatic residues in a 92-atom ring and is one of the largest synthetic organic macrocycles to be crystallographically characterized. The synthesis, structure, and ring expansion of a related macrocyclic ether-ketone are also reported. PMID- 11735578 TI - Synthesis of 3,4-disubstituted isoquinolines via palladium-catalyzed cross coupling of o-(1-alkynyl)benzaldimines and organic halides. AB - [reaction: see text] 3,4-Disubstituted isoquinolines have been prepared in good yields by the palladium-catalyzed cross-coupling of N-tert-butyl-o-(1 alkynyl)benzaldimines with aryl, allylic, and alkynyl halides. PMID- 11735579 TI - Spirocyclic restriction of nucleosides. Synthesis of the first exemplary syn-1 oxaspiro[4.4]nonanyl member. AB - [reaction: see text] The potential benefits associated with the spirocyclic restriction of nucleosides are summarized. Following exploration of a pi allylpalladium route to 5'-alpha- or syn-dideoxy examples, we evaluated MOM protection of the 5'-hydroxyl as being suited to the synthesis of the first member of this new class of nucleoside mimic. PMID- 11735580 TI - Spirocyclic restriction of nucleosides. An analysis of protecting group feasibility while accessing prototype anti-1-oxaspiro[4.4]nonanyl mimics. AB - [reaction: see text] The first spirocyclic nucleoside featuring a beta-hydroxyl (anti) at C5' has yielded to synthesis. While the OMOM functionality proved to be sensitive to the conditions necessary to incorporate heterocyclic bases, PMB protection of the carbinol was readily accommodated. The remarkably similar minimum-energy conformations of the title compounds relative to natural thymidine as deduced by Amber calculations in the gas phase are noted. PMID- 11735582 TI - Novel conversion of 1,2-disubstituted cis-epoxides to one-carbon homologated allylic alcohols using dimethylsulfonium methylide. AB - [reaction: see text] The treatment of cis-epoxides with an excess of dimethylsulfonium methylide affords one-carbon homologated allylic alcohols in good to excellent yields. PMID- 11735581 TI - The first naturally occurring Tie2 kinase inhibitor. AB - [structure: see text] Bioassay-guided fractionation of the plant Acacia aulacocarpa, guided by a bioassay for Tie2 tyrosine kinase activity, yielded the novel triterpene 3,21-dioxo-olean-18-en-oic acid (1) as the first naturally occurring non-protein inhibitor of Tie2 kinase. The structure of 1 was assigned by analysis of spectral data. In addition to its activity as an inhibitor of Tie2 kinase, compound 1 also shows modest activity against a variety of cultured mammalian cells. PMID- 11735583 TI - Homolytic carbostannylation of alkenes and alkynes with tributylstannyl enolates. AB - In the presence of AIBN, tributylstannyl enolates derived from aromatic ketones reacted with electron-deficient alkenes and a variety of alkynes to give the corresponding carbostannylated adducts. The reactions with methyl acrylate gave alpha-tributylstannylmethyl-gamma-ketoesters, unlike the known Michael-type reaction of stannyl enolates forming delta-ketoesters. The carbostannylation of alkynes proceeded in an anti addition mode to afford beta,gamma-unsaturated ketones. The reactivity of stannyl enolates as radical transfer agents could be utilized for radical cyclization of 1,6-enynes. PMID- 11735584 TI - Lactone-derived carbon-centered radicals: formation and reactivity with oxygen. AB - [reaction: see text] Several lactones were examined to test the reactivity of carbon-centered radicals toward oxygen. Notably, the radical derived from 2 coumaranone (4) is unreactive toward oxygen, while 2-cuomaranone itself shows enhanced reactivity toward hydrogen abstraction by alkoxyl radicals. We propose that five parameters influence diminished reactivity toward oxygen, i.e., (a) benzylic resonance stabilization, (b) unpaired spin delocalization on oxygen, (c) favorable stereoelectronic effects, (d) electron-withdrawing effects, and (e) steric effects. PMID- 11735585 TI - First generation design, synthesis, and evaluation of azepine-based cryptophycin analogues. AB - [structure: see text] Azepine-based cryptophycin mimics (+)-4 and (+)-5 have been designed and synthesized. Biological evaluation revealed modest in vitro activity against several human tumor cell lines, thereby supporting the utility of novel scaffolds for the design and synthesis of cryptophycin analogues. PMID- 11735586 TI - Visual recognition of triamines by phenolphthalein derivatives: consideration of the structure of the colored complex. AB - [structure: see text] A hybrid molecule 1 consisting of phenolphthalein and two crown ether moieties can be used to discriminate the length of linear triamines strictly by color development. The purple color is developed most deeply at -10 degrees C and fades with either an increase or decrease in temperature. PMID- 11735588 TI - A stereoselective synthesis of the C10-C31 (BCDEF ring) portion of pinnatoxin A. AB - [reaction: see text] An efficient synthesis of the C10-C31 (BCDEF ring) portion of pinnatoxin A has been achieved. The key step is a highly stereoselective construction of the dispiroketal (BCD ring) system employing an intramolecular hetero-Michael reaction of a reversibly formed hemiketal alkoxide through the use of LiOMe. PMID- 11735587 TI - Visual enantiomeric recognition using chiral phenolphthalein derivatives. AB - [structure: see text] Optically active artificial host molecules 2-5 based on a phenolphthalein skeleton have been prepared for visual enantiomeric recognition of alanine derivatives 8 and 9. The receptor 3 discriminates (R)-8 and (R)-9 from (S)-8 and (S)-9, respectively, to develop a purple color. PMID- 11735589 TI - A rapid access to biaryl ether containing macrocycles by pairwise use of Ugi 4CR and intramolecular S(N)Ar-based cycloetherification. AB - [reaction: see text] From readily accessible starting materials, macrocycles with an endo aryl-aryl ether bond are synthesized in only two operations by combination of the Ugi four-component reaction and an intramolecular S(N)Ar reaction. The nitro group serves as an activator for the macrocyclization and provides a handle for the introduction of functional group diversity. A Ugi reaction promoted by ammonium chloride in aprotic solvent is documented for the first time. PMID- 11735590 TI - BINOL-based diphosphonites as ligands in the asymmetric Rh-catalyzed conjugate addition of arylboronic acids. AB - [reaction: see text] BINOL-based diphosphonites having achiral backbones are useful ligands in the Rh-catalyzed conjugate addition of arylboronic acids to alpha,beta-unsaturated carbonyl compounds. The nature of the achiral backbone determines the direction and degree of enantioselectivity, with er values of up to 99.5:0.5 possible. PMID- 11735591 TI - N-Cyclopropylimine-1-pyrroline rearrangement. A novel photochemical reaction. AB - [reaction: see text] A novel aza-(vinylcyclopropane-cyclopentene) photochemical rearrangement is reported. 1-Pyrrolines are easily synthesized in good yields from N-cyclopropylimines. Hydrogen, alkyl, and aryl groups can be placed anywhere within the system, and the reaction proceeds regiospecifically. PMID- 11735593 TI - Generation, observation, and free radical reactivity of aliphatic bisketenes: the solution to a long-standing problem. AB - [reaction: see text] Bisketenes O=C=CH(CH(2))(n)()CH=C=O (1b,c,d, n = 4, 3, 6) and (E)-O=C=CHCH=CHCH=C=O (E-13) were generated in solution by dehydrochlorination of bis(acyl chlorides) and by photochemical Wolff rearrangements and identified by their characteristic IR signals. The bisketenes react with aminoxyl radicals to give tetraaddition products for 1b and conjugate 1,6-diaddition for E-13. PMID- 11735592 TI - An intramolecular [2 + 3] cycloaddition route to fused 5-heterosubstituted tetrazoles. AB - [reaction: see text] Fused 5-heterotetrazole ring systems are synthesized in high yield via intramolecular [2 + 3] cycloadditions of organic azides and heteroatom substituted nitriles. Cyanates, thiocyanates, and cyanamides are all competent dipolarophiles for this reaction. A variety of scaffolds are tolerated when the new enclosed ring is five- or six-membered. PMID- 11735594 TI - Lipase/palladium-catalyzed asymmetric transformations of ketoximes to optically active amines. AB - [reaction: see text] Prochiral ketoximes were asymmetrically transformed to optically active amines in the acetylated forms by coupled lipase/palladium catalysis in the presence of an acyl donor under 1 atm of hydrogen. PMID- 11735595 TI - Palladium-catalyzed cross-coupling reaction of triarylbismuths with aryl halides and triflates. AB - [reaction: see text] Palladium-catalyzed cross-coupling reaction of triarylbismuths with aryl bromides, iodides, and triflates proceeded efficiently in the presence of K(2)CO(3) or CsF. PMID- 11735596 TI - Coupling of alkynylTMS derivatives with vinylic iodides. An efficient route to 1,3-enynes and dienes. AB - [reaction: see text] CuCl-promoted coupling of alkynylTMS derivatives with vinyl iodides leads to 1,3-enynes in high yield. Enynes prepared from homopropargylic alcohols undergo intramolecular hydrosilylation and subsequent silyl cleavage with TBAF to afford 1,3-dienes. PMID- 11735597 TI - Efficient enantioselective hydrosilylation of ketones catalyzed by air stable copper fluoride-phosphine complexes. AB - [reaction: see text] Copper(II) fluoride-chiral diphosphine systems catalyze the hydrosilylation of several ketones with moderate to excellent enantioselectivities. An oxygen acceleration effect was observed and led to a practical protocol with low catalyst loading. PMID- 11735598 TI - Chain extension of sugar delta-lactones with the enolate of tert-butyl bromoacetate and elaboration into functionalized C-ketosides, C-glycosides, and C glucosyl glycines. AB - [structure: see text] We describe the synthesis of a series of exocyclic sugar epoxides 1 prepared in a one-step procedure from sugar delta-lactones with the enolate of tert-butyl bromoacetate. Ring opening of the sugar oxiranes provides C ketosides while reduction affords functionalized C-glycosides bearing an alpha hydroxy ester moiety. The alpha-hydroxy ester can be converted into C-glucosyl glycine analogues 2. PMID- 11735599 TI - [2,2]paracyclophane-based N,O-ligands in alkenylzinc additions to aldehydes. AB - [reaction: see text] The application of planar and central chiral [2,2]paracyclophane-based N,O-ligands in asymmetric alkenylzinc additions to various aldehydes is described, which gives rise to very high ee's especially for difficult substrates. A fine-tuning of the alkenylzinc species by employing different transmetalation reagents is reported, allowing control of the steric bulk of the alkenylzinc species and thus the selectivity of the catalysis. PMID- 11735600 TI - Aerobic organocatalytic photoinitiated arene oxidations: application to electron transfer initiated cyclization reactions. AB - [reaction: see text] Electron transfer initiated oxidative cyclization reactions can be effected by catalytic amounts of N-methylquinolinium hexafluorophosphate through photoirradiation in the presence of dioxygen. Solid sodium thiosulfate serves as an effective reducing agent to remove the reactive oxygen species formed from catalyst regeneration and radical coupling reactions with O(2), allowing these cyclizations to proceed efficiently on preparatively useful scales. PMID- 11735603 TI - Individualising aminoglycoside dosage regimens after therapeutic drug monitoring: simple or complex pharmacokinetic methods? AB - Measurements of aminoglycoside concentration in serum are used to individualise dosage regimens (dose per administration and/or administration interval) with the goal of attaining the desired therapeutic range as quickly as possible. Therapeutic range is defined in terms of peak concentration (to monitor effectiveness) and trough concentration (to avoid toxicity). This article focuses on methods to individualise aminoglycoside dosage regimens in the context of extended dosage intervals. Simple pharmacokinetic methods involve linear dosage adjustment based on peak or trough concentrations or area under the concentration time curve, or nomograms. The once daily aminoglycoside nomogram determines the dosage interval for aminoglycosides given as a fixed dose per administration, based on a single concentration measurement drawn 6 to 14 hours after the start of the first infusion. This is a preferred method because of its simplicity, strong pharmacodynamic rationale and prospective validation in a large population. However, it does not work when the fixed dose assumed is not relevant, for example for patients with burns, cystic fibrosis, ascites or pregnancy. Furthermore, it has not been validated in children. In these cases, a more sophisticated method is required. Complex pharmacokinetic methods require dedicated software. Non-Bayesian least-squares methods allow the optimisation of both the dose and the dosage interval, but require aminoglycoside concentrations from two or more samples taken in the post-distributive phase during a single dosage interval. With Bayesian least-squares methods, only one concentration measurement is required, although any number of samples can be taken into account. In the Bayesian maximum a posteriori (MAP) method, the parameter estimates are taken as the values corresponding to the maximum of the posterior density. In 'full' Bayesian approaches (also called stochastic control), all the information about the parameters revealed by the posterior distribution is taken into account, and the optimal regimen is found by minimising the expected value of the weighted sum of squared deviations between predicted and target concentrations. If the population model is reasonably well known, Bayesian methods (MAP or stochastic control) should be used because of their good predictive performance. Although only one concentration measurement is required, better precision is afforded by a two-sample strategy, preferably drawn 1 and 6 hours after the start of the first infusion. If the population model is not known, then the non-Bayesian least-squares method is the method of choice, because of its robustness and lack of requirement for prior information about the distribution of parameters in the population. PMID- 11735602 TI - Pharmacogenetics: the therapeutic drug monitoring of the future? AB - Genetic variability in drug response occurs as a result of molecular alterations at the level of drug-metabolising enzymes, drug targets/receptors, and drug transport proteins. In this paper, we discuss the possibility that therapeutic drug monitoring (TDM) in the future will involve not the mere measurement and interpretation of drug concentrations but will include both traditional TDM and pharmacogenetics-oriented TDM. In contrast to traditional TDM, which cannot be performed until after a drug is administered to the patient. pharmacogenetics oriented TDM can be conducted even before treatment begins. Other advantages of genotyping over traditional TDM include, but are not limited to, the following: (i) it does not require the assumption of steady-state conditions (or patient compliance) for the interpretation of results; (ii) it can often be performed less invasively (with saliva, hair root or buccal swab samples); (iii) it can provide predictive value for multiple drugs [e.g. a number of cytochrome P450 (CYP) 2D6, CYP2C 19 or CYP2C9 substrates] rather than a single drug; (iv) it provides mechanistic, instead of merely descriptive, information; and (v) it is constant over an individual's lifetime (and not influenced by concurrent drug administration, alteration in hormonal levels or disease states). Pharmacogenetic information can be applied a priori for initial dose stratification and identification of cases where certain drugs are simply not effective. However, traditional TDM will still be required for all of the reasons that we use it now. In current clinical practice, pharmacogenetic testing is performed for only a few drugs (e.g. mercaptopurine, thioguanine, azathioprine, trastuzumab and tacrine) and in a limited number of teaching hospitals and specialist academic centres. We propose that other drugs (e.g. warfarin, phenytoin, codeine, oral hypoglycaemics, tricyclic antidepressants, aminoglycosides, digoxin, cyclosporin, cyclophosphamide, ifosfamide, theophylline and clozapine) are potential candidates for pharmacogenetics-oriented TDM. However, prospective studies of phaymacogenetics-oriented TDM must be performed to determine its efficacy and cost effectiveness in optimising therapeutic effects while minimising toxicity. In the future, in addition to targeting a patient's drug concentrations within a therapeutic range, pharmacists are likely to be making dosage recommendations for individual drugs on the basis of the individual patient's genotype. As we enter the era of personalised drug therapy, we will be able to identify not only the best drug to be administered to a particular patient, but also the most effective and safest dosage from the outset of therapy. PMID- 11735604 TI - Pharmacokinetics of coagulation factors: clinical relevance for patients with haemophilia. AB - Haemophilia is a recessively inherited coagulation disorder, in which an X chromosome mutation causes a deficiency of either coagulation factor VIII (FVIII) in haemophilia A, or factor IX (FIX) in haemophilia B. Intravenous administration of FVIII or FIX can be used to control a bleeding episode, to provide haemostasis during surgery or for long term prophylaxis of bleeding. In special cases, activated factor VII (FVIIa) may be used instead of FVIII or FIX. The aim of this work is to review the pharmacokinetics of FVIII, FIX and FVIIa and to give an outline of the use of pharmacokinetics to optimise the treatment of patients with haemophilia. The pharmacokinetics of FVIII are well characterised. The systemic clearance (CL) of FVIII is largely determined by the plasma level of von Willebrand factor (vWF), which protects FVIII from degradation. Typical average CL in patients with normal vWF levels is 3 ml/h/kg, with an apparent volume of distribution at steady state (Vss) that slightly exceeds the plasma volume of the patient, and the average elimination half-life (t1/2) is around 14 hours. There are still some discrepancies in the literature on the pharmacokinetics of FIX. The average CL of plasma-derived FIX seems to be 4 ml/h/kg, the Vss is 3 to 4 times the plasma volume and the elimination t1/2 often exceeds 30 hours. FVIIa has a much higher CL (average of 33 ml/h/kg), and a short terminal t1/2 (at 2 to 3 hours). The Vss is 2 to 3 times the plasma volume. Since the therapeutic levels of coagulation factors are well defined in most clinical situations, applied pharmacokinetics is an excellent tool to optimise therapy. Individual tailoring of administration in prophylaxis has been shown to considerably increase the cost effectiveness of the treatment. Dosage regimens for the treatment of bleeding episodes or for haemostasis during surgery are also designed using pharmacokinetic data, and the advantages of using a constant infusion instead of repeated bolus doses have been explored. The influence of antibodies (inhibitors) on the pharmacokinetics of FVIII and FIX is in part understood, and the doses of coagulation factor needed to treat a patient can tentatively be calculated from the antibody titre. In conclusion, therapeutic monitoring of coagulation factor levels and the use of clinical pharmacokinetics to aid therapy are well established in the treatment of patients with haemophilia. PMID- 11735606 TI - Quantitative pharmacogenetics of nortriptyline: a novel approach. AB - OBJECTIVE: To quantitatively model nortriptyline clearance as a function of the cytochrome P450 (CYP) 2D6 genotype and to estimate the contribution of genotype to the interindividual variability in steady-state plasma concentration and metabolic clearance. DESIGN: Modelling study using data from two previously published studies. PARTICIPANTS: 20 healthy volunteers receiving single oral doses of nortriptyline and 20 patients with depression on steady-state oral treatment. METHODS: A total of 275 nortriptyline plasma concentrations were analysed by standard nonlinear regression and nonlinear mixed effect models. The pharmacokinetic model was a 1-compartment model with first order absorption and elimination. All participants had previously been genotyped with respect to the CYP2D6 polymorphism. RESULTS: A model in which the intrinsic clearance is a linear function of the number of functional CYP2D6 genes and hepatic blood flow is fixed to 60 L/h gave the closest fit of the pharmacokinetic model to the data. Stable estimates were obtained for population pharmacokinetic parameters and interindividual variances. Assuming 100% absorption, the model allows systemic clearance and bioavailability to be estimated. Bioavailability was found to vary between 0.17 and 0.71, depending on the genotype. Using the frequency distribution of CYP2D6 genotype with the above results we estimate that, in compliant Swedish individuals on nortriptyline monotherapy, the number of functional CYP2D6 genes could explain 21% of the total interindividual variance in oral clearance of nortriptyline and 34% of that in steady-state plasma concentrations. CONCLUSION: Nonlinear mixed-effects modelling can be used to quantify the influence of the number of functional CYP2D6 genes on the metabolic clearance and plasma concentration of drugs metabolised by this enzyme. Gene dose has a significant impact on drug pharmacokinetics and prior knowledge of it may aid in predicting plasma concentration of the drug and thus tailoring patient specific dosage regimens. PMID- 11735607 TI - Utilisation of pharmacokinetic-pharmacodynamic modelling and simulation in regulatory decision-making. AB - Modelling and simulation (M&S) play an important role in regulatory decision making that affects both the public and industry. Technological advances in various fields related to drug development call for more focus on ways to optimise current drug development practices. Recognition of the potential of M&S by regulatory agencies inevitably has a substantial impact on drug development. The objective of the current review is to present the various regulatory initiatives for application of M&S to clinical drug development. The relevant parts of the various recommendations issued by the US Food and Drug Administration (FDA), via guidance documents and advisory committee meeting proceedings, are highlighted. Application of M&S to a variety of activities, such as integrating pharmacokinetic-pharmacodynamic knowledge across a new drug application and designing efficient trials, is discussed. Some of the challenges that pharmaceutical institutions currently face when implementing M&S projects, such as team structure, communication with regulators, training and time constraints, are also presented, and solutions are proposed. PMID- 11735608 TI - Clinical pharmacokinetics of non-nucleoside reverse transcriptase inhibitors. AB - Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are a diverse group of compounds that induce allosteric changes in the human immunodeficiency virus type 1 (HIV-1) reverse transcriptase, thus rendering the enzyme incapable of converting viral RNA to DNA. Unlike nucleoside analogue inhibitors of reverse transcriptase, NNRTIs do not require sequential phosphorylation to elicit antiretroviral activity. There are currently 3 approved NNRTIs: nevirapine, delavirdine and efavirenz. Although possessing a common mechanism of action, these agents can be differentiated by both molecular and pharmacokinetic characteristics. Each of the NNRTIs is metabolised to some degree by the cytochrome P450 (CYP) system of enzymes, making them prone to clinically significant drug interactions. In addition, they elicit variable effects on other medications, acting as either inducers or inhibitors of drugs metabolised by CYP. These drug interactions are an important consideration in the clinical use of these agents as a part of combination antiretroviral therapy. Additional factors such as the influence of food and pH on oral absorption, and protein binding, must also be considered. PMID- 11735610 TI - Renal elimination of amikacin and the aging process. AB - OBJECTIVE: Although amikacin is primarily eliminated via glomerular filtration, drug concentrations are not consistently predicted in all patients. To better describe the relationship between amikacin clearance and both age and renal function, we used a new heuristic approach involving statistical analysis of dependence. DESIGN AND SETTING: Retrospective pharmacokinetic study using data from seven centres in France. PARTICIPANTS: 634 patients with sepsis aged between 18 and 98 years of age who received intravenous amikacin. METHODS: Clearance of amikacin was modelled using the NonParametric EM algorithm for a two-compartment model (NPEM2) with intravenous infusion. RESULTS: A total of 2499 serum amikacin determinations was available for analysis. The relationship between the clearance of amikacin and age was weak. Interestingly, the Z method, which filters data based on dependence criteria, selected data that were best fitted by a polynomial function (r = 0.90; p < 0.001). This representation of the polynomial function was similar to a previously proposed theoretical model describing covariations between the clearance of amikacin and age. However, the polynomial function applied to only 33% of the patients that were selected by the Z method. The correlation between the clearance of amikacin and renal function was also relatively low (r = 0.39). The Z method exhibited a continuous and strong dependence pattern between the clearance of amikacin and age for 49% of the patients. CONCLUSIONS: The Z methodology, which filters data using dependence criteria, confirms that age, renal function and amikacin clearance are strongly related, but only in less than half of a large sample of patients with sepsis without renal pathology. These results suggest that other variables should be taken into account in order to improve the description of the behaviour of amikacin. The Z methodology improved the classical description of relationships between variables, and should be applied to better select pertinent variables in pharmacokinetic studies. PMID- 11735605 TI - Pharmacokinetic aspects of treating infections in the intensive care unit: focus on drug interactions. AB - Pharmacokinetic interactions involving anti-infective drugs may be important in the intensive care unit (ICU). Although some interactions involve absorption or distribution, the most clinically relevant interactions during anti-infective treatment involve the elimination phase. Cytochrome P450 (CYP) 1A2, 2C9, 2C19, 2D6 and 3A4 are the major isoforms responsible for oxidative metabolism of drugs. Macrolides (especially troleandomycin and erythromycin versus CYP3A4), fluoroquinolones (especially enoxacin, ciprofloxacin and norfloxacin versus CYP1A2) and azole antifungals (especially fluconazole versus CYP2C9 and CYP2C19, and ketoconazole and itraconazole versus CYP3A4) are all inhibitors of CYP mediated metabolism and may therefore be responsible for toxicity of other coadministered drugs by decreasing their clearance. On the other hand, rifampicin is a nonspecific inducer of CYP-mediated metabolism (especially of CYP2C9, CYP2C19 and CYP3A4) and may therefore cause therapeutic failure of other coadministered drugs by increasing their clearance. Drugs frequently used in the ICU that are at risk of clinically relevant pharrmacokinetic interactions with anti-infective agents include some benzodiazepines (especially midazolam and triazolam), immunosuppressive agents (cyclosporin, tacrolimus), antiasthmatic agents (theophylline), opioid analgesics (alfentanil), anticonvulsants (phenytoin, carbamazepine), calcium antagonists (verapamil, nifedipine, felodipine) and anticoagulants (warfarin). Some lipophilic anti-infective agents inhibit (clarithromycin, itraconazole) or induce (rifampicin) the transmembrane transporter P-glycoprotein, which promotes excretion from renal tubular and intestinal cells. This results in a decrease or increase, respectively, in the clearance of P-glycoprotein substrates at the renal level and an increase or decrease, respectively, of their oral bioavailability at the intestinal level. Hydrophilic anti-infective agents are often eliminated unchanged by renal glomerular filtration and tubular secretion, and are therefore involved in competition for excretion. Beta-lactams are known to compete with other drugs for renal tubular secretion mediated by the organic anion transport system, but this is frequently not of major concern, given their wide therapeutic index. However, there is a risk of nephrotoxicity and neurotoxicity with some cephalosporins and carbapenems. Therapeutic failure with these hydrophilic compounds may be due to haemodynamically active coadministered drugs, such as dopamine, dobutamine and furosemide, which increase their renal clearance by means of enhanced cardiac output and/or renal blood flow. Therefore, coadministration of some drugs should be avoided, or at least careful therapeutic drug monitoring should be performed when available. Monitoring may be especially helpful when there is some coexisting pathophysiological condition affecting drug disposition, for example malabsorption or marked instability of the systemic circulation or of renal or hepatic function. PMID- 11735609 TI - Delivery of neurotrophic factors to the central nervous system: pharmacokinetic considerations. AB - Neurotrophic factors are proteins with considerable potential in the treatment of central nervous system (CNS) diseases and traumatic injuries. However, a significant challenge to their clinical use is the difficulty associated with delivering these proteins to the CNS. Neurotrophic factors are hydrophilic, typically basic, monomeric or dimeric proteins, mostly in the size range of 5 to 30 kDa. Neurotrophic factors potently support the development, growth and survival of neurons, eliciting biological effects at concentrations in the nanomolar to femtomolar range. They are not orally bioavailable and the blood brain and blood-cerebrospinal fluid barriers severely limit their ability to enter into and act on sites in the CNS following parenteral systemic routes of administration. Most neurotrophic factors have short in vivo half-lives and poor pharmacokinetic profiles. Their access to the CNS is restricted by rapid enzymatic inactivation, multiple clearance processes, potential immunogenicity and sequestration by binding proteins and other components of the blood and peripheral tissues. The development of targeted drug delivery strategies for neurotrophic factors will probably determine their clinical effectiveness for CNS conditions. Achieving significant CNS target site concentrations while limiting systemic exposure and distribution to peripheral sites of action will lessen unwanted pleiotropic effects and toxicity. Local introduction of neurotrophic factors into the CNS intraparenchymally by direct injection/infusion or by implantation of delivery vectors such as polymer matrices or genetically modified cells yields the highest degree of targeting, but is limited by diffusion restrictions and invasiveness. Delivery of neurotrophic factors into the cerebrospinal fluid (CSF) following intracerebroventricular or intrathecal administration is less invasive and allows access to a much wider area of the CNS through CSF circulation pathways. However, diffusional and cellular barriers to penetration into surrounding CNS tissue and significant clearance of CSF into the venous and lymphatic circulation are also limiting. Unconventional delivery strategies such as intranasal administration may offer some degree of CNS targeting with minimal invasiveness. This review presents a summary of the neurotrophic factors and their indications for CNS disorders, their physicochemical characteristics and the different approaches that have been attempted or suggested for their delivery to the CNS. Future directions for further research such as the potential for CNS disease treatment utilising combinations of neurotrophic factors, displacement strategies, small molecule mimetics, chimaeric molecules and gene therapy are also discussed. PMID- 11735611 TI - Bacterial meningitis: current controversies in approaches to treatment. AB - Acute bacterial meningitis continues to be a disease with unacceptably high mortality and morbidity rates in both adults and children worldwide, despite advances in antibacterial therapy. Death or permanent disability occurs frequently. The causative organism varies with age, immune function and immunisation status. Infection with Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae type b (Hib) is associated with the majority of cases, with Listeria monocytogenes and Streptococcus agalactiae being more prevalent pathogens at the extremes of age (<3 months or >50 years). Antibacterial resistance is an increasing problem, particularly in pneumococcal bacteria but increasingly in other organisms. The increasing prevalence of resistance of pneumococcus to penicillin and the cephalosporins complicates therapy and may have an important impact on treatment outcome. Increased understanding of the pathophysiology has allowed advances in diagnosis and therapy. The use of adjunctive corticosteroids remains controversial, but is probably beneficial in reducing neurological sequelae in children. In adults the evidence is less clear. Vaccination has virtually eradicated Hib meningitis in some countries. Recent introduction of a conjugate vaccine against serogroup C meningococci in the UK has caused a dramatic reduction in the incidence of invasive disease due to this organism. A 7-valent pneumococcal vaccine promises a similar reduction in the incidence of invasive pneumococcal disease. In the meantime, the emergence of widespread resistance of organisms to antibacterial agents, in particular among the common organisms causing bacterial meningitis, remains the biggest challenge in therapy. PMID- 11735612 TI - Magnesium for neuroprotection in ischaemic stroke: rationale for use and evidence of effectiveness. AB - Magnesium exhibits a range of neuronal and vascular actions that may ameliorate ischaemic CNS insults, including stroke. Significant neuroprotection with magnesium has been observed in different models of focal cerebral ischaemia in many laboratories, with infarct volume reductions between 25 and 61%. Maximal neuroprotection is evident at readily attainable serum concentrations, and neuroprotection is still seen when administration is delayed up to 6 hours after onset of ischaemia. Clinical use of magnesium in pre-eclampsia and acute myocardial infarction confirms its safety and tolerability. Five small trials in acute stroke have reported reduced odds of death or dependence with administration of magnesium, but confidence intervals are wide, and definitive data from ongoing large trials are awaited. PMID- 11735613 TI - Diagnosis and treatment of nonconvulsive status epilepticus. AB - Nonconvulsive status epilepticus (SE) is not uncommon and comprises at least one third of all cases of SE. However, nonconvulsive SE consists of very different syndromes, a common feature being the difficulty in making the diagnosis. In this review, nonconvulsive SE is divided into typical absence SE, complex partial SE, nonconvulsive SE in patients with learning difficulties (including electrical SE during sleep, atypical absence SE and tonic SE), and nonconvulsive SE in coma. These conditions have different prognoses and treatments. The diagnosis of these conditions is critically dependent on EEG. When the EEG demonstrates typical ictal patterns, the diagnosis is usually straightforward. However, in many circumstances the EEG has to be differentiated from encephalopathic patterns, and this differentiation can prove troublesome, although the clinical and electrographic response to treatment can prove helpful. Nonconvulsive SE in patients with learning difficulties possibly provides the greatest diagnostic difficulty; the clinical presentation can be subtle resulting in the diagnosis being frequently missed. Whether the neuronal damage that occurs in convulsive SE and in animal models of limbic SE also occurs in nonconvulsive SE in humans is still a matter of debate. There are critical differences between the animal models and the human condition. Indeed, the prognosis of nonconvulsive SE is usually dependent on the underlying aetiology rather than the persistence of electrographic discharges. Because of these doubts, a more conservative approach to the treatment of particular types of nonconvulsive SE (those with a better prognosis) has been taken in this article. Thus, in most instances, oral benzodiazepines for the treatment of typical absence SE and complex partial SE are recommended. In some circumstances intravenous medication is necessary, but in neither condition is anaesthetic coma recommended. This contrasts with nonconvulsive SE in coma in which a more aggressive approach is suggested. Until there are more relevant animal models, and controlled trials of conservative versus more aggressive treatment, treatment regimens for nonconvulsive SE will remain largely speculative. PMID- 11735614 TI - Third-generation antidepressants: do they offer advantages over the SSRIs? AB - Third-generation antidepressants are a group of antidepressant agents of variable action, not confined to serotonin reuptake inhibition. These agents include venlafaxine, reboxetine, nefazodone and mirtazapine. Claims have been made for these agents in terms of improved efficacy, faster speed of onset of effect and greater safety in the treatment of depression compared with previous medications, such as the selective serotonin reuptake inhibitors (SSRIs). This article reviews the evidence for these improvements. Thirty active comparator studies were reviewed involving the third-generation antidepressant agents. While there were isolated reports of improvements over comparator agents for venlafaxine, reboxetine and mirtazepine, there were no convincing differences between third generation agents and comparators in terms of overall efficacy, relapse prevention and speed of onset. The third-generation antidepressants were, however, of equivalent safety to SSRIs and maintained improvements in safety over first-generation agents. PMID- 11735616 TI - Frovatriptan. AB - black triangle Frovatriptan, a new serotonin receptor agonist developed for the acute treatment of migraine, has high affinity for serotonin 5-HT1B and 5-HT1D receptor subtypes and is a potent stimulator of contraction in human basilar arteries. black triangle A long terminal elimination half-life (approximately 26 hours) is a distinctive pharmacokinetic feature of frovatriptan which appears to be independent of dose, age, gender and renal function. black triangle A single oral dose of frovatriptan 2.5mg was effective in the acute treatment of migraine providing meaningful relief within 2 hours to approximately twice as many recipients as placebo in clinical trials. black triangle Consistent relief of migraine symptoms was achieved in patients who treated a number of consecutive attacks with frovatriptan and the incidence of 24-hour migraine recurrence was reduced. black triangle Frovatriptan was well tolerated in clinical trials, with the overall incidence of adverse events occurring with frovatriptan 2.5mg only slightly higher than that reported with placebo. Mild to moderate fatigue, nausea and paraesthesia were the most commonly reported drug-related adverse events. PMID- 11735615 TI - Future prospects for the drug treatment of epilepsy. AB - Great progress has been made in the last 150 years in the pharmacological management of epilepsy, and, despite the increasing number of technological advances available, antiepileptic drugs (AEDs) remain the mainstay of treatment for the vast majority of patients with epilepsy. This review looks at possible avenues of development in the drug treatment of epilepsy. The strengths and weaknesses of those AEDs which are currently licensed are examined, and ways in which their use may be improved are discussed (e.g. rational combinations, use of new formulations). Potentially new targets that may allow the development of effective treatments are highlighted (neuroimmunological manipulation, decreasing inherent drug resistance mechanisms, and modification of adenosine neurotransmission), and a summary of the most promising AEDs currently in development is provided [e.g. carabersat, ganaxolone, harkoseride, MDL 27192, safinamide (NW 1015), pregabalin, retigabine, talampanel, valrocemide, losigamone and BIA 2093]. PMID- 11735619 TI - Potential and current use of N-methyl-D-aspartate (NMDA) receptor antagonists in diseases of aging. AB - The N-methyl-D-aspartate (NMDA) receptor complex is a subtype of glutamate receptor and its dysfunction is involved in many neurological disorders associated with aging, including chronic pain, depression, stroke and Parkinson's disease. Multiple clinical trials using NMDA receptor antagonists have been aborted mainly due to the severe psychomimetic adverse effects of these drugs that occur before concentrations can reach an adequate level in the brain. In this review, we present the evidence that clinically safer NMDA antagonists such as memantine and nitroglycerin, and the combination drug nitro-memantine, are promising as drugs in treating neurodegenerative diseases. PMID- 11735620 TI - Current management strategies for hepatitis B in the elderly. AB - Despite the availability of an efficient vaccine, chronic hepatitis B virus (HBV) infection remains a major public health problem worldwide. The World Health Organization estimates that there are still 350 million chronic carriers of the virus who are at risk of developing chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). Antiviral therapy consists of the administration of either interferon-alpha (IFN alpha) or lamivudine. In the elderly, specific issues should be addressed. Because of the long duration of viral infection, screening for HCC is warranted in these patients, as new therapeutic options are being developed. Antiviral treatment for chronic hepatitis B is indicated in patients with elevated transaminases, the presence of HBV replication, and inflammatory activity on liver histology analysis, providing the patient has no other serious health problem impacting on life expectancy. Since IFN alpha therapy may cause many general adverse effects, lamivudine may be the best current treatment option in this patient population. The pharmacokinetics of lamivudine in the elderly are slightly different from those in younger adults but this does not require dose adjustment, except in the presence of renal function impairment. However, the beneficial effects of lamivudine therapy must be weighed against the selection of drug-resistant mutants. New therapeutic strategies are now under evaluation and may be available in the future for the elderly population. Besides mass HBV vaccination programmes, people sharing a house with patients infected with HBV should be vaccinated to prevent viral transmission. PMID- 11735622 TI - Immunosuppression in elderly renal transplant recipients: are current regimens too aggressive? AB - Renal transplantation is an accepted and successful treatment modality in elderly patients with end-stage renal disease. In comparison with maintenance dialysis, transplantation has been shown to confer a mortality benefit as well as improvements in quality of life in older individuals with end-stage renal disease. Despite this, overall outcomes of renal transplantation in elderly individuals have, in general, been less successful than those of younger renal transplant recipients. Largely, this has been due to the particular vulnerability of elderly patients to the immunosuppressive medications used in renal transplantation. This review article covers these issues in some detail and briefly discusses some of the pharmacokinetic, pharmacodynamic, physiological and immunological differences between younger and older transplant recipients. Elderly renal transplant recipients have both a higher rate of patient death and allograft loss censored for death. Upon multivariate analysis, age of the recipient is strongly associated with allograft loss independent of other known factors. Acute rejections are less frequent in older individuals; however the consequence of a rejection if it occurs is negative for long-term graft survival. On the other hand, death by infection is vastly increased in older versus younger renal transplant recipients. In general, the pharmacokinetics of the immunosuppressive agents are little affected by age, but the tolerance to these agents seems to decrease with increasing age. Elderly renal transplant recipients present a very difficult clinical challenge. As the elderly become an ever increasing segment of the renal transplant population, new and innovative immunosuppressive strategies will have to be considered and applied. PMID- 11735621 TI - Management of painful diabetic neuropathy. AB - Type 2 diabetes mellitus is a prevalent disease in the US which affects more than 15 million people. As the disease progresses over time, neuropathic pain can become a common complication; it is present in more than 50% of individuals with diabetes mellitus aged >60 years. The pathogenesis of diabetic neuropathy is theorized to be multifactorial. Numerous medications, some with different mechanisms of action, have been examined regarding their effects on the symptoms associated with diabetic neuropathy such as pain, paraesthesia and numbness. However, the majority of the studies have included small patient populations. Tricyclic antidepressants, amitriptyline and desipramine in particular, have been relatively well studied and shown to be effective. However, anticholinergic adverse effects may limit their usefulness and may preclude use in the elderly. Studies have also shown gabapentin to be effective and well tolerated in the treatment of diabetic neuropathy. Capsaicin cream provides another treatment option with a favourable adverse effect profile. Many other medications have been evaluated in diabetic neuropathy; however, more placebo-controlled studies with adequate patient populations need to be performed to solidify their role in treatment. PMID- 11735623 TI - Epidemiology of breast cancer in older women: implications for future healthcare. AB - Breast cancer in elderly women is already a significant public health problem. Elderly women have a 6-fold higher breast cancer incidence rate and 8-fold higher mortality rate compared with non-elderly women. Because of demographic trends, the number of elderly women diagnosed with breast cancer is likely to increase substantially in the coming decades. Specifically, if incidence rates remain constant, we project a 72% increase in the number of elderly women in the US diagnosed with breast cancer by 2025, an increase from approximately 89,500 in 1998 to almost 154,000 in 2025. If this projection holds true, the sheer magnitude of the increase in patients has profound implications for the delivery of medical care. Considerable planning is needed to ensure that the infrastructure is in place to effectively treat these patients. The burgeoning number of elderly patients with breast cancer accentuates the need for more definitive evidence concerning preventing and treating breast cancer in the elderly. Treatment patterns for elderly patients with breast cancer have been shown to differ from those for non-elderly patients, but the evidence base for differentiating treatment plans by age is deficient. For example, information is needed to tease apart the relative importance of age per se compared with important age-related factors, such as comorbidity. Patient care will benefit from an interdisciplinary team approach that includes oncologists, geriatricians, surgeons, radiation oncologists, nurses and social workers. The continued increase in life expectancy necessitates well-crafted strategies for the primary and secondary prevention of breast cancer. Carefully addressing the priorities for breast cancer prevention and control in the elderly during the first portion of the century may reap substantial dividends by the end of the century. PMID- 11735624 TI - Fetal tissue transplantation for patients with Parkinson's disease: a database of published clinical results. AB - Over the past 13 years approximately 300 patients with Parkinson's disease have received transplants of human fetal dopamine cells in an attempt to reduce or control disease symptoms. Many of these patients have had improvements in their motor skills and a reduction in their daily levodopa administration. However, improvements are far from guaranteed and questions need to be answered before this technique can be widely applied. To help address some of these issues, a search of all the published results of patients with Parkinson's disease transplanted with human fetal tissue was conducted. This generated a database of 70 transplant recipients who had their levodopa administration and clinical benefit reported both prior to transplant and at least 6 months post-transplant. Furthermore, the number of years of disease onset prior to transplant was available for all recipients. This database was examined for motor improvement and reduction in levodopa dosage for up to 2 years post-transplant to determine the effects of time on transplant outcome. The database showed that most recipients had significant improvements in motor skills and levodopa administration, and that most benefits were observed in the first 6 months post transplant. In addition, the database demonstrated that the number of years of disease onset prior to transplantation was not a predictor of patient outcome 1 year post-transplant. Current and future directions in fetal tissue transplantation research and replacements for fetal tissue are discussed. PMID- 11735628 TI - Imatinib: A Viewpoint by John M. Goldman. PMID- 11735625 TI - Cisplatin/epinephrine injectable gel. AB - 1. Cisplatin/epinephrine injectable gel is a preparation for intratumoural injection containing cisplatin 4 mg/ml, epinephrine (adrenaline) 0.1 mg/ml and bovine collagen as a protein carrier matrix. It has been evaluated for the palliative treatment of accessible inoperable metastatic or recurrent solid tumours. 2. The vasoconstrictor action of epinephrine limits the diffusion of cisplatin into the systemic circulation. Intratumoural injection of cisplatin/epinephrine injectable gel achieves high concentrations of cisplatin in the tumour with very low concentrations in plasma and other tissues. 3. In double blind randomised trials, cisplatin/epinephrine injectable gel was more efficacious than placebo in the palliative treatment of recurrent and resistant head and neck squamous cell carcinoma. 4. Cisplatin/epinephrine injectable gel reduced tumour size and improved local symptoms in patients with metastatic breast cancer, metastatic malignant melanoma, oesophageal carcinoma and hepatic tumours in a number of noncomparative clinical trials. 5. Adverse events with the use of cisplatin/epinephrine injectable gel are mainly limited to the local site of injection. No systemic adverse events such as nephrotoxicity, neurotoxicity or ototoxicity have been reported with use of this preparation. PMID- 11735629 TI - Imatinib: A Viewpoint by Brian J. Druker. PMID- 11735630 TI - Pancreatic islet cell transplantation: likely impact on current therapeutics for type 1 diabetes mellitus. AB - Pancreatic islet transplantation as a cell-based treatment for diabetes mellitus has been pursued for over quarter of a century, ever since the first successful use of this approach to cure diabetes mellitus in rodents in the 1970s. However, even though autoislet intrahepatic transplantation in patients with chronic pancreatitis who did not have diabetes mellitus was successful in the 1980s, reliable success with alloislet transplantation in patients with diabetes mellitus remained elusive until the year 2000. The reasons for previous failures appear to include use of corticosteroids as an immunosuppressive agent, a drug that both interferes with islet beta-cell function and promotes resistance to insulin action. Corticosteroid-free immunosuppressive regimens in one recent series have permitted much greater success for over 1 year. Benefits accrued in this series include normal fasting glucose levels, normal levels of glycosylated haemoglobin, and total independence from exogenous insulin or other therapy for hyperglycaemia. These dramatic results have presented the research community with the challenges of replicating these results and, ultimately, with providing solutions to the serious supply and demand issues that will inevitably follow. PMID- 11735633 TI - Responsible prescribing for upper respiratory tract infections. AB - Upper respiratory tract infections (URTIs) are responsible for a large amount of community antibacterial use worldwide. Recent systematic reviews have demonstrated that most URTIs resolve naturally, even when bacteria are the cause. The high consumer expectation for antibacterials in URTIs requires intervention by the general practitioner and a number of useful strategies have been developed. Generic strategies, including eliciting patient expectations, avoiding the term 'just a virus', providing a value-for-money consultation, providing verbal and written information, empowering patients, conditional prescribing, directed education campaigns, and emphasis on symptomatic treatments, should be used as well as discussion of alternative medicines when relevant. The various conditions have differing rates of bacterial infection and require different approaches. For acute rhinitis, laryngitis and tracheitis, viruses are the only cause and, therefore, antibacterials are never required. In acute sore throat (pharyngitis) Streptococcus pyogenes is the only important bacterial cause. A scoring system can help to increase the likelihood of distinguishing a streptococcal as opposed to viral infection, or alternatively patients should be given antibacterials only if certain conditions are fulfilled. Strategies for treating acute otitis media vary in different countries. Most favour the strategy of prescribing antibacterials only when certain criteria are fulfilled, delaying antibacterial prescribing for at least 24 hours. In otitis media with effusion, on the other hand, there is no primary role for antibacterials, as the condition resolves naturally in almost all patients aged >3 months. Detailed strategies for acute sinusitis have not been worked out but restricting antibacterial prescribing to certain clinical complexes is currently recommended by several authorities because of the high natural resolution rate. PMID- 11735631 TI - Underutilisation of ACE inhibitors in patients with congestive heart failure. AB - Congestive heart failure (CHF) is associated with substantial morbidity and mortality, and is the only major cardiovascular disease increasing in prevalence. Despite abundant evidence to support their efficacy and cost-effectiveness, angiotensin-converting enzyme (ACE) inhibitors are sub-optimally used in patients with CHF. This paper reviews the evidence for the sub-optimal use of ACE inhibitors in patients with CHF, the factors contributing to this, and its implications for health systems. A systematic review of all articles assessing practice patterns (specifically the use of ACE inhibitors in CHF) identified by MEDLINE, search of bibliographies, and contact with content experts was undertaken. 37 studies have documented the use of ACE inhibitors in patients with CHF. Studies assessing use among all patients with CHF document 33% to 67% (median 51%) of all patients discharged from hospital and 10% to 36% (median 26%) of community dwelling patients were prescribed ACE inhibitors. Rates of ACE inhibitor use range from 43% to 90% (median of 71%) amongst those discharged from hospital having known systolic dysfunction, and from 67% to 95% (median of 86%) for those monitored in specialty clinics. Moreover, the dosages used in the 'real world' are substantially lower than those proven efficacious in randomised, controlled trials, with evaluations reporting only a minority of patients achieving target doses and/or an overall mean dose achieved to be less than one half of the target dose. Factors predicting the use and optimal dose administration of ACE inhibitors are identified, and include variables relating to the setting (previous hospitalisation, specialty clinic follow-up), the physician (cardiology specialty versus family practitioner or general internist, board certification), the patient (increased severity of symptoms, male, younger), and the drug (lower frequency of administration). In light of the substantial evidence for reductions in morbidity and mortality, clearly, the prescription of ACE inhibitors is sub-optimal. Wide variability in ACE inhibitor use is noted, with higher rates consistently reported among patients having systolic dysfunction confirmed by an objective assessment--an apparent minority of the those having CHF. Optimisation of the prescription of proven efficacious therapies has the potential to confer a substantial reduction in the total cost of care for patients with CHF by reducing hospitalisations and lengths of hospital stays. It is likely that only multifaceted programs targeted toward the population at large will yield benefits to the healthcare system, given the widespread nature of the sub-optimal prescription of therapies proven effective in the management of patients with CHF. PMID- 11735632 TI - The use of silymarin in the treatment of liver diseases. AB - The high prevalence of liver diseases such as chronic hepatitis and cirrhosis underscores the need for efficient and cost-effective treatments. The potential benefit of silymarin (extracted from the seeds of Silybum marianum or milk thistle) in the treatment of liver diseases remains a controversial issue. Therefore, the objective of this review is to assess the clinical efficacy and safety of silymarin by application of systematic approach. 525 references were found in the databases, of which 84 papers were retained for closer examination and 36 were deemed suitable for detailed analysis. Silymarin has metabolic and cell-regulating effects at concentrations found in clinical conditions, namely carrier-mediated regulation of cell membrane permeability, inhibition of the 5 lipoxygenase pathway, scavenging of reactive oxygen species (ROS) of the R-OH type and action on DNA-expression, for example, via suppression of nuclear factor (NF)-kappaB. Pooled data from case record studies involving 452 patients with Amanita phalloides poisoning show a highly significant difference in mortality in favour of silibinin [the main isomer contained in silymarin] (mortality 9.8% vs 18.3% with standard treatment; p < 0.01). The available trials in patients with toxic (e.g. solvents) or iatrogenic (e.g. antispychotic or tacrine) liver diseases, which are mostly outdated and underpowered, do not enable any valid conclusions to be drawn on the value of silymarin. The exception is an improved clinical tolerance of tacrine. In spite of some positive results in patients with acute viral hepatitis, no formally valid conclusion can be drawn regarding the value of silymarin in the treatment of these infections. Although there were no clinical end-points in the four trials considered in patients with alcoholic liver disease, histological findings were reported as improved in two out of two trials, improvement of prothrombin time was significant (two trials pooled) and liver transaminase levels were consistently lower in the silymarin-treated groups. Therefore, silymarin may be of use as an adjuvant in the therapy of alcoholic liver disease. Analysis was performed on five trials with a total of 602 patients with liver cirrhosis. The evidence shows that, compared with placebo, silymarin produces a nonsignificant reduction of total mortality by 4.2% [odds ratio (OR) 0.75 (0.5 - 1.1)]; but that, on the other hand, the use of silymarin leads to a significant reduction in liver-related mortality of-7% [OR: 0.54 (0.3 - 0.9); p < 0.01]. An individual trial reported a reduction in the number of patients with encephalopathy of -8.7% (p = 0.06). In one study of patients with cirrhosis-related diabetes mellitus, the insulin requirement was reduced by -25% (p < 0.01). We conclude that available evidence suggests that silymarin may play a role in the therapy of (alcoholic) liver cirrhosis. Silymarin is has a good safety record and only rare case reports of gastrointestinal disturbances and allergic skin rashes have been published. This review does not aim to replace future prospective trials aiming to provide the 'final' evidence of the efficacy of silymarin. PMID- 11735634 TI - Premenstrual asthma: epidemiology, pathogenesis and treatment. AB - The exacerbation of asthma in the premenstrual period has long been of interest. Premenstrual asthma has been estimated to affect up to 40% of females with asthma, although the exact prevalence of this phenomenon is unclear as studies have involved small numbers in hospital clinics. Large-scale community-based studies are required to estimate its true prevalence. Researchers are slowly piecing together clues as to the aetiology and pathogenesis of the disorder. Female sex-steroid hormones play an important role but the exact mechanism is still unknown. Recent evidence suggests that increased airway hyperresponsiveness, an indicator of underlying airway inflammation, during the luteal phase of the menstrual cycle may account for premenstrual exacerbations. In addition, there is now evidence of impaired or altered beta2-adrenoceptor function and regulation in females with asthma, which may have a part to play. Accurate diagnosis is dependent on a detailed history and the demonstration of premenstrual dip in peak expiratory flow. Exacerbations in the majority of women will respond to the usual treatment of bronchial asthma. However, a few women will experience significant morbidity or treatment-related adverse effects. Case reports suggest that the combined oral contraceptive pill or gonadotrophin releasing hormone analogues may be effective in these patients. This requires substantiation by randomised controlled trials. PMID- 11735635 TI - Pyelonephritis in pregnancy: treatment options for optimal outcomes. AB - Acute pyelonephritis is one of the most common indications for antepartum hospitalisation. When acute pyelonephritis is diagnosed, conventional treatment includes intravenous fluid and parenteral antibacterial administration. There are limited data by which to assess the superiority of one antibacterial regimen over the other in terms of efficacy, patient acceptance and safety for the developing fetus. There is a small body of evidence to support the ambulatory treatment of pregnant women with pyelonephritis in the first and early second trimesters. PMID- 11735637 TI - Darbepoetin Alfa: A Viewpoint by Allen R. Nissenson. PMID- 11735636 TI - Darbepoetin alfa. AB - Darbepoetin alfa is a novel erythropoiesis-stimulating protein developed for the treatment of anaemia associated with chronic kidney disease. In single-dose studies in patients undergoing dialysis, the mean terminal half-life for intravenous darbepoetin alfa was approximately 3-fold longer than for intravenous recombinant human erythropoitin (r-HuEPO, epoetin alfa; 25.3 vs 8.5 hours). The mean terminal half-life after subcutaneous administration of darbepoetin alfa was 48.8 hours. In randomised nonblind trials in patients undergoing dialysis, darbepoetin alfa (0.45 pg/kg) given once weekly for the correction of anaemia increased haemoglobin (Hb) levels to a similar extent as darbepoetin alfa three times weekly or r-HuEPO two or three times weekly. A double-blind, randomised clinical trial reported that switching patients from a three-times weekly regimen of r-HuEPO to once weekly darbepoetin alfa with additional placebo twice weekly (all intravenously) maintained Hb levels between 9.0 and 13.0 g/dl to a similar extent as continued treatment with r-HuEPO three times weekly. In a randomised nonblind study, r-HuEPO-naive patients with chronic renal insufficiency received either subcutaneous darbepoetin alfa once weekly or r-HuEPO twice weekly. 93% of patients receiving darbepoetin alfa and 92% of patients receiving r-HuEPO achieved a Hb increase of > or = 1.0 g/dl from baseline and the mean increase in Hb level over the initial 4 weeks was similar for both treatments. The number and frequency of adverse events, withdrawals and deaths reported in clinical trials did not differ between patients receiving darbepoetin alfa and patients receiving r-HuEPO. There have been no reports of immune responses to darbepoetin alfa in 1534 patients receiving treatment for up to 2 years. PMID- 11735638 TI - Darbepoetin Alfa: A Viewpoint by Anatole Besarab. PMID- 11735639 TI - Darbepoetin Alfa: A Viewpoint by Iain C. Macdougall. PMID- 11735641 TI - Orlistat: A Viewpoint by Priscilla A. Hollander. PMID- 11735640 TI - Orlistat: in the prevention and treatment of type 2 diabetes mellitus. AB - Orlistat is a nonsystemically acting gastric and pancreatic lipase inhibitor that limits the absorption of dietary fat. A retrospective pooled analysis of three 2 year, double-blind, randomised, placebo-controlled trials involving patients with obesity revealed that orlistat recipients were more likely to experience an improvement, and less likely to experience a deterioration, in glucose tolerance status than placebo recipients. In comparison with placebo, orlistat recipients had significantly greater reductions in glycosylated haemoglobin and fasting plasma glucose levels in large, double-blind, randomised, placebo-controlled studies of 24 to 52 weeks' duration involving patients with obesity and type 2 diabetes mellitus. In one such study, the dosage of concomitant sulphonylureas was able to be reduced in more orlistat than placebo recipients (43.2 vs 28.9%), with discontinuation of sulphonylurea therapy achieved in 11.7% of orlistat recipients. The most common adverse effects reported in orlistat recipients with type 2 diabetes mellitus relate to the gastrointestinal system and are similar to those reported in studies involving patients without type 2 diabetes mellitus. PMID- 11735642 TI - Orlistat: A Viewpoint by Hans Hauner. PMID- 11735643 TI - Amisulpride: a review of its use in the management of schizophrenia. AB - Amisulpride, a substituted benzamide derivative, is a second-generation (atypical) antipsychotic. At low doses, it enhances dopaminergic neurotransmission by preferentially blocking presynaptic dopamine D2/D3 autoreceptors. At higher doses, amisupride antagonises postsynaptic dopamine D2 and D3 receptors, preferentially in the limbic system rather than the striatum, thereby reducing dopaminergic transmission. In patients with acute exacerbations of schizophrenia, the recommended dosage of amisulpride is 400 to 800 mg/day, although dosages < or =1200 mg/day may be administered. In comparative trials, amisulpride administered within this range (400 to 1200 mg/day) was as effective as haloperidol 5 to 40 mg/day, flupenthixol 25 mg/day and risperidone 8 mg/day in patients with acute exacerbations of schizophrenia with predominantly positive symptoms. Amisulpride was more effective than haloperidol but equally effective as risperidone in controlling negative symptoms. Amisulpride 400 to 800 mg/day was more effective than haloperidol, risperidone and flupenthixol in controlling affective symptoms in these patients. In randomised, double-blind trials involving patients with predominantly negative symptoms of schizophrenia, amisulpride 50 to 300 mg/day was more effective than placebo. Amisulpride is effective as maintenance therapy in patients with chronic schizophrenia. Long term treatment with amisulpride was associated with improvements in quality of life and social functioning. Amisulpride is generally well tolerated. In well controlled trials, the neurological tolerability profile (including ratings on extrapyramidal symptom scales) of amisulpride 400 to 1200 mg/day was superior to that of the conventional antipsychotics (haloperidol or flupenthixol), but was similar to that of the atypical antipsychotic risperidone. At low dosages of amisulpride (< or =300 mg/day), the incidence of adverse events (including extrapyramidal symptoms) reported with amisulpride was similar to that with placebo. CONCLUSION: In comparative trials, amisulpride 400 to 1200 mg/day showed efficacy in reducing overall symptomatology and positive symptoms similar to that of conventional antipsychotics and newer atypical antipsychotics in patients with acute exacerbations of schizophrenia. Moreover, its effective alleviation of negative and affective symptoms, its lower association with extrapyramidal symptoms and loss of cognitive function than conventional antipsychotics and its long-term efficacy justifies consideration of the use of higher dosages of amisulpride in this group of patients. Consequently, the dosage of amisulpride that is recommended in patients with acute exacerbations of schizophrenia is 400 to 800 mg/day, although dosages < or =1200 mg/day may be administered. Lower dosages of amisulpride (50 to 300 mg/day) should be considered for the management of patients with negative symptoms of schizophrenia. Amisulpride is a first-line treatment option in the management of schizophrenia in the acute phase and for the maintenance of treatment response. PMID- 11735644 TI - Post-marketing surveillance and vigilance for medical devices: the European approach. AB - The extent to which the medical device manufacturers are responsible for actively monitoring the performance of their products after they have successfully passed the rigorous pre-market approval process has always been a matter of diverse opinion. Within Europe, the law is unhelpfully vague on this point. While there are some comparatively clear obligations for reporting incidents to the authorities (known as the 'vigilance system'), little detail is given on how diligently the manufacturer should try to find out about such incidents. In the early stages of the European Community Directives covering medical devices, there was much emphasis upon formulating guidance to help interpret the vigilance reporting requirements. It is, however, only recently that attention has turned to attempting to clarify what is expected from post-marketing surveillance (PMS) in its broader sense. This article discuses both the vigilance and PMS processes and outlines the currently available European, and particularly UK, guidance documents which are aimed at promoting a more level playing field across industry where these activities are concerned. In particular, it explains the principle differences between vigilance and post-marketing surveillance: the former being the reporting of adverse incidents by manufacturers to the regulatory authorities and their subsequent sharing of key incident data between each other; the latter being the process by which information on overall devise performance is captured, analysed and acted upon. Nevertheless, it is still a struggle to gain widespread appreciation that these two activities are not in fact one and the same. PMID- 11735645 TI - Hepatotoxicity with thiazolidinediones: is it a class effect? AB - Decreased insulin sensitivity plays a major role in various human diseases. particularly type 2 diabetes mellitus, and is associated with a higher risk of atherosclerosis and cardiovascular complications. Thiazolidinediones, more commonly termed glitazones, are the first drugs to specifically target muscular insulin resistance. They have proven efficacy for reducing plasma glucose levels in patients with type 2 diabetes mellitus treated with diet alone, sulphonylureas, metformin or insulin. In addition, they are associated with some improvement of the cardiovascular risk profile. However, troglitazone, the first compound approved by the Food and Drug Administration in the US, proved to be hepatotoxic and was withdrawn from the market after the report of several dozen deaths or cases of severe hepatic failure requiring liver transplantation. It remains unclear whether or not hepatotoxicity is a class effect or is related to unique properties of troglitazone. Rosiglitazone and pioglitazone, two other glitazones, appear to have similar efficacy with regard to blood glucose control in patients with type 2 diabetes mellitus as compared with troglitazone. In controlled clinical trials, the incidence of significant (> or =3 x upper limit of normal) increases in liver enzyme levels (ALT in particular) was similar with rosiglitazone or pioglitazone as compared with placebo, whereas troglitazone was associated with a 3-fold greater incidence. In contrast to the numerous case reports of acute liver failure in patients receiving troglitzone, only a few case reports of hepatotoxicity have been reported in patients treated with rosiglitazone until now, with a causal relationship remaining uncertain. Furthermore, no single case of severe hepatotoxicity has been reported yet with pioglitazone. It should be mentioned that troglitazone, unlike pioglitazone and rosiglitazone, induces the cytochrome P450 isoform 3A4, which is partly responsible for its metabolism, and may be prone to drug interactions. Importantly enough, obesity, insulin resistance and type 2 diabetes mellitus are associated with liver abnormalities, especially non-alcoholic steatohepatitis, independent of any pharmacological treatment. This association obviously complicates the selection of patients who are good candidates for a treatment with glitazones as well as the monitoring of liver tests after initiation of therapy with any thiazolidinedione compound. While regular monitoring of liver enzymes is still recommended and more long term data are desirable, current evidence from clinical trials and postmarketing experience in the US supports the conclusion that rosiglitazone and pioglitazone do not share the hepatotoxic profile of troglitazone. PMID- 11735646 TI - Drug-induced congenital defects: strategies to reduce the incidence. AB - Approximately 1% of congenital anomalies relate to pharmacological exposure and are. in theory, preventable. Prevention consists of controlled administration of drugs known to have teratogenic properties (e.g. retinoids, thalidomide). When possible, prevention could take the form of the use of alternative pharmacological therapies during the pre-conception period for certain specific pathologies, selecting the most appropriate agent for use during pregnancy [e.g. haloperidol or a tricyclic antidepressant instead of lithium; anticonvulsant drug monotherapy in place of multitherapy; propylthiouracil instead of thiamazole (methimazole)], and substitution with the most suitable therapy during pregnancy (e.g. insulin in place of oral antidiabetics; heparin in place of oral anticoagulants; alpha-methyldopa instead of ACE inhibitors). Another strategy is the administration of drugs during pregnancy taking into account the pharmacological effects in relation to the gestation period (e.g. avoidance of chemotherapy during the first trimester, avoidance of nonsteroidal anti inflammatory drugs in the third trimester, and avoidance of high doses of benzodiazepines in the period imminent to prepartum). PMID- 11735647 TI - A comparison of liposomal formulations of doxorubicin with drug administered in free form: changing toxicity profiles. AB - The anthracycline antibiotic doxorubicin has wide activity against a number of human neoplasms and is used extensively both as a single agent and in combination regimens. In addition to the use of free, unencapsulated doxorubicin, there are two US Food and Drug Administration approved liposomal formulations of doxorubicin currently available, with several additional liposomal formulations being researched either in the laboratory or in clinical trials. The two approved liposomal formulations of doxorubicin have significantly different lipid compositions and loading techniques, which lead to both unique pharmacokinetic and toxicity profiles, distinct from those of the unencapsulated form. This article discusses the toxicities associated with the free form of doxorubicin, as well as those associated with the two most common liposomal formulations, namely Doxil and Myocet. One of the key toxicity issues linked to the use of free doxorubicin is that of both an acute and a chronic form of cardiomyopathy. This is circumvented by the use of liposomal formulations, as these systems tend to sequester the drug away from organs such as the heart, with greater accumulation in liver, spleen and tumours. However, as will be discussed, the liposomal formulations of doxorubicin are not without their own related toxicities, and, in the case of Doxil, may be associated with the unique toxicity of palmar-plantar erythrodysaesthesia. Overall, the use of liposomal doxorubicin allows for a greater lifetime cumulative dose of doxorubicin to be administered, however acute maximal tolerated doses differ significantly, with that of Myocet being essentially equivalent to free doxorubicin, while higher doses of Doxil may be safely administered. This review highlights the differences in both toxicity and pharmacokinetic properties between free doxorubicin and the different liposomal formulations, as have been determined in pre-clinical and clinical testing against a number of different human neoplasms. The need for further testing of the liposomal formulations prior to the replacement of free doxorubicin with liposomal doxorubicin in any established combination therapy regimens, as well as in combination with the newer therapeutics such as monoclonal antibodies is also discussed. PMID- 11735648 TI - Medication overuse headache: a focus on analgesics, ergot alkaloids and triptans. AB - Medication overuse headache (MOH, formerly known as drug-induced headache) is a well known disorder following the frequent use of analgesics or any other antiheadache drug including serotonin 5-HT(1B/D) agonists (triptans). Recent studies suggest clinical features of MOH depend on the substance class that has been overused. The delay between the frequent intake of any antiheadache drug and daily headache is shortest for triptans (mean 1.7 years), longer for ergot alkaloids (mean 2.7 years) and longest for analgesics (mean 4.9 years). Treatment includes withdrawal followed by structured acute therapy and initiation of specific prophylactic treatment for the underlying primary headache. The relapse rate within 6 months after successful withdrawal is about 30% and increases steadily up to 50% after 5 years. PMID- 11735649 TI - Long term follow-up studies of users of nonprescription medicines purchased from community pharmacies: some methodological issues. AB - BACKGROUND: Despite the wider availability of medicines to the general public, little is known about their safety when supplied without prescription. Pilot work has already tested 4 methods of recruiting users of ibuprofen purchased from community pharmacies. This paper describes the piloting of a fifth method (a shortened questionnaire), long term follow-up rates of all methods, consistency of reporting of ibuprofen use, and issues relating to possible comparison groups in pharmacovigilance studies. METHODS: A shortened version of a previously tested recruitment questionnaire was used. Eligible study participants were all users, aged over 17 years, purchasing ibuprofen from a research network of community pharmacies (n = 61) in Grampian, Scotland. Postal questionnaires were sent at I week and 2, 6 and 12 months irrespective of the method of recruitment. The follow up questionnaires collected information about ibuprofen and other drug usage, symptoms and associated health service utilisation. RESULTS: The shortened form recruited 67% of people issued with a questionnaire. The overall 12-month follow up rate was 67%, although there were important differences in the rates by method of recruitment. There was reasonable consistency in the reporting of use or non use of ibuprofen at different follow-up times. In the 12 months after the index purchase, 17% of participants never used any ibuprofen (non-users) and 28% used it for more than 8 weeks in total (long term users). At 12 months, long term users were significantly more likely than short term users (< or =8 weeks total use) or non-users to have experienced dizziness, skin rash, itchy skin and wheeziness in the previous week. CONCLUSIONS: Our pilot work has confirmed the feasibility of recruiting, and following-up over prolonged periods, users of nonprescription medicines. Evidence of long term use of ibuprofen confirms the need for pharmacovigilance studies of this drug, although further work is required to identify a suitable comparison group in order to inform the interpretation of such investigations. PMID- 11735650 TI - Measles vaccination and inflammatory bowel disease: controversy laid to rest? AB - The increasing incidence of Crohn's disease has lead to speculation about changes in exposures to environmental or infectious agents. Considerable attention has focused on the role of measles infection and/or vaccination in the pathogenesis of Crohn's disease and ulcerative colitis. Current evidence regarding the association between measles vaccination and inflammatory bowel disease (IBD) comprises analytic epidemiological studies, a case-series report and ecological studies. The first of these, a 1995 cohort study, found an association between measles vaccination and Crohn's disease and ulcerative colitis, but was widely questioned on methodological grounds. This was followed by a 1997 case-control study showing no association between measles vaccination and IBD. In 1998, public concern was rekindled by a report of 12 children with nonspecific colitis, ileal lymphoid-nodular hyperplasia, and developmental disorders largely attributed to measles-mumps-rubella vaccine, but the nature of the report limited its scientific conclusions. Two additional studies, one case-control and one cohort, then followed and neither found an association with measles vaccination. Of the several ecological studies of measles vaccine coverage or measles schedule changes, none found an association with rates of IBD. The role of measles infection in IBD has been examined more extensively with studies of in utero measles exposure, measles infection early in life, and laboratory based investigations. An initial report of high rates of Crohn's disease among pregnancies affected by measles infection was followed by negative studies. Numerous case-control and ecological studies of children with measles infections early in life have also had discordant findings. Of three recent cohort studies, two showed no relationship between infection with early measles exposure and risk for IBD, while one found an approximate 3-fold elevation in risk. Laboratory investigations into persistent measles infection and IBD have been contentious. While some investigators have claimed to find persistent measles infection among patients with IBD, others, using highly sensitive polymerase chain reaction techniques, have not been able to replicate the findings. Recent controversy has centred on whether there is any evidence for molecular mimicry in the pathogenesis of IBD. In summary, available evidence does not support an association between measles-containing vaccines and risk of IBD, nor between measles infection and IBD. While further research is necessary into the causal factors underlying Crohn's disease and ulcerative colitis, continued public education efforts are needed to reassure the public about vaccine safety and to prevent declines in vaccine coverage. PMID- 11735652 TI - Methods and systems to detect adverse drug reactions in hospitals. AB - Detection of adverse drug reactions (ADRs) in hospitals offers the chance to detect serious ADRs resulting in hospitalisation and ADRs occurring in hospitalised patients, i.e. patients with high comorbidity and receiving drugs that are administered only in hospitals. The most commonly applied methods involve stimulated spontaneous reporting of doctors and nurses, comprehensive collection by trained specialists and, more recently, computer-assisted approaches using routine data from hospital information systems. The different methods of ADR detection used result in different rates and types of ADRs and, consequently, in different drug classes being responsible for these ADRs. Another factor influencing the results of surveys is the interpretation of the term ADR, where some authors adhere to the strict definition of the World Health Organization and many others include intended and unintended poisoning as well as errors in prescribing and dispensing, thus referring to adverse drug events. Depending on the method used for screening of patients, a high number of possible ADRs and only few definite ADRs are found, or vice versa. These variations have to be taken into account when comparing the results of further analyses performed with these data. ADR rates and incidences in relation to the number of drugs prescribed or patients exposed have been calculated in only a few surveys and projects, and this interesting pharmacoepidemiological approach deserves further study. In addition, the pharmacoeconomic impact of ADRs, either resulting in hospitalisation or prolonging hospital stay, has been estimated using different approaches. However, a common standardised procedure for such calculations has not yet been defined. Although detection of ADRs in hospitals offers the opportunity to detect severe ADRs of newly approved drugs, these ADRs are still discovered by spontaneous reporting systems. The prospects offered by electronic hospital information systems as well as implementation of pharmacoepidemiological approaches increases the possibilities and the value of ADR detection in hospitals. PMID- 11735651 TI - Therapeutic drug monitoring databases for postmarketing surveillance of drug-drug interactions. AB - Drug-drug interactions can be associated with patient morbidity due to either increased toxicity or a potentially ineffective concentration. Because interactions cannot always be anticipated during drug development and actual patients receiving a drug for therapeutic use often differ from those included in clinical trials, postmarketing surveillance is essential. Therapeutic drug monitoring (TDM) databases offer a unique opportunity in this respect. Prerequisites for TDM databases to provide valid information in a pharmacoepidemiological perspective include the following: precise description of exposure to the potentially interacting drugs; measurement of parent compound and active metabolites through accurate and precise analytical techniques; documentation of relevant patient characteristics that may act as confounding factors (e.g. gender, age, smoking habits); repeated assessments over time if possible; and sound pharmacokinetic framework for data selection, analysis and interpretation. The contribution of TDM to the documentation of drug-drug interactions takes advantage of different possible study designs, discussed on the basis of recently published studies. The single case report plays an important role as an alert signal. It is illustrated for a patient on long-term treatment, who displayed an unexpectedly high clozapine concentration after the introduction of ciprofloxacin comedication. The prospective on and off comedication panel study shows advantages in terms of carefully selected inclusion criteria and control of treatment modalities. A study of the thioridazine-fluvoxamine interaction is presented, with patients followed on thioridazine monotherapy, after introduction of fluvoxamine and after its discontinuation. The main advantage of the retrospective large-scale TDM database screen is representativeness of patients actually treated, whereas drawbacks are related to quality of data and suitability for valid interpretation. Such an approach is illustrated by a review of data collected over 10 years of routine TDM that allowed documenting induction of nortriptyline metabolism by carbamazepine and inhibition by several phenothiazines. Finally, population pharmacokinetics is well suited to observational data collected for TDM purpose, provided quality is ascertained. Focus is placed on interindividual variability and relationship between pharmacokinetic parameters and patient characteristics, including comedication. The population approach is discussed with respect to a study that documented a 32% increase of haloperidol clearance associated with anticonvulsant comedication, in addition to effects of age and bodyweight. Among factors to consider for improved effectiveness in the use of TDM databases for postmarketing surveillance of drug-drug interactions, integration of efficacy and safety data in future studies and communication of expert recommendations to prescribing physicians are essential. PMID- 11735653 TI - Bodyweight gain and anticonvulsants: a comparative review. AB - Bodyweight gain is a common and frequent undesirable effect associated with the use of anticonvulsant drugs. This has been observed for many years with valproic acid (sodium valproate) and carbamazepine, and also, more recently, with some of the newer anticonvulsants such as vigabatrin and gabapentin. Very often bodyweight gain in children, adolescents and adults with epilepsy taking such anticonvulsants results in cosmetic adverse effects. On the other hand, bodyweight gain is disturbing to general health, with a possible increase in the risk of diabetes mellitus or heart disease. Other potential adverse effects, such as the association of obesity with polycystic ovaries, have been reported with the use of valproic acid. Potential mechanisms of anticonvulsant-associated bodyweight gain are not yet clear and differ between drugs used. The involvement of lowered blood glucose level, which may stimulate eating through an effect on the hypothalamus, constitutes one of the possible mechanisms. Lowered blood glucose levels may result from a competition between the binding of the drug and long chain fatty acids. An increased availability of the latter stimulates insulin production and lowers the serum glucose levels. Another possible explanation for lowered blood glucose may be a deficiency in carnitine directly caused by the drug, that would result in a reduction of fatty acid metabolism and an increase in glucose consumption. An enhancing effect of gamma-aminobutyric acid-mediated neurotransmission may increase appetite for carbohydrates and reduce energy expenditure. An antidiuretic hormone-like effect or effects on norepinephrine (noradrenaline) or serotonin-mediated neurotransmission are more rarely considered. Many studies on anticonvulsant-associated bodyweight gain illustrate how we could better define the risk factors for the development of anticonvulsant-induced bodyweight gain and uncover the mechanisms behind it. PMID- 11735654 TI - Risk-benefit assessment of glatiramer acetate in multiple sclerosis. AB - Glatiramer acetate, formerly known as copolymer 1, is a mixture of synthetic polypeptides composed of four amino acids. Glatiramer acetate has been shown to be effective in preventing and suppressing experimental autoimmune encephalitis (EAE), the animal model of multiple sclerosis (MS). Therefore it was tested in several clinical studies, where it was found to slow the progression of disability and to reduce the relapse rate and the magnetic resonance imaging (MRI)-defined disease activity and burden in relapsing-remitting MS. As a daily standard dose, 20mg of glatiramer acetate is injected subcutaneously. After injection, glatiramer acetate undergoes rapid degradation to amino acids and shorter peptides; so it is not possible to measure any systemic plasma concentrations or excretion rates. Two major mechanisms have been proposed to explain the effects of glatiramer acetate in EAE and MS: the induction of glatiramer acetate-reactive T helper 2 (Th2)-like regulatory suppressive cells and the interference with T cell activation as an altered peptide ligand. The most common adverse effects were mild injection site reactions (erythema, inflammation and induration). The most remarkable adverse event is the acute and transient immediate postinjection reaction manifested by flushing, chest tightness, palpitations and dyspnoea. Other reported adverse effects are transient chest pain and lymphadenopathy. Antibodies to glatiramer acetate induced during treatment do not interfere with its clinical effects. In several controlled clinical studies, glatiramer acetate has been shown to provide consistent, reproducible clinical benefits in the target population of patients with relapsing-remitting MS. The safety profile and risk-benefit ratio are excellent. Overall, glatiramer acetate is very well tolerated and has an excellent risk-benefit profile in patients with relapsing-remitting MS. PMID- 11735656 TI - Lipid abnormalities induced by novel antipsychotic drugs. PMID- 11735658 TI - Technologies to improve immunisation safety. AB - Ever since a vaccine was first used against smallpox, adverse events following immunisation have been reported. Adverse reactions may be caused by a fault in vaccine production, idiosyncratic responses or unsafe handling and vaccine administration practices. Technological advances that promise to bypass many of the dangers currently associated with vaccine administration are described. Plans for the next decade and beyond include developing injection-free systems for vaccine delivery that overcome the limitations of current immunisation programmes and help prevent programmatic mistakes. Also under development are new parenteral administration devices such as the auto-disable syringe and the mono-dose pre filled device, and mucosal and transcutaneous immunisation systems. Training needs to be at the forefront of efforts to limit human error. Above all, there must be a willingness to respond to new climates and new technologies in order to ensure safe immunisation of children globally. PMID- 11735655 TI - Treating epilepsy in the elderly: safety considerations. AB - The incidence of epilepsy increases with advancing age. Epilepsy in the elderly has different aetiologies from that in younger populations, cerebrovascular disease being the most common condition associated with seizures. Partial seizures are the predominant seizure type in older patients. A diagnosis of epilepsy in the elderly is based mainly on the history and is frequently delayed. In addition, seizure imitators are especially frequent. In many cases ancillary tests for diagnosis may show normal age-related variants, sometimes making results difficult to interpret. Treating epilepsy in the elderly is problematic due to a number of issues that relate to age and comorbidity. The physical changes associated with increasing age frequently lead to changes in the pharmacokinetics of many anticonvulsants. The treatment of epilepsy in the elderly is also complicated by the existence of other diseases that might affect the metabolism or excretion of anticonvulsants and the presence of concomitant medications that might interact with them. Moreover, specific trials of anticonvulsants in the aged population are scarce. General guidelines for treatment include starting at lower doses, slowing the titration schedule, individualising the choice of anticonvulsant to the characteristics of the patient, avoiding anticonvulsants with important cognitive or sedative adverse effects, and where possible, treating with monotherapy. PMID- 11735659 TI - Benefits and risks of self medication. AB - Self medication is becoming an increasingly important area within healthcare. It moves patients towards greater independence in making decisions about management of minor illnesses, thereby promoting empowerment. Self medication also has advantages for healthcare systems as it facilitates better use of clinical skills, increases access to medication and may contribute to reducing prescribed drug costs associated with publicly funded health programmes. However, self medication is associated with risks such as misdiagnosis, use of excessive drug dosage, prolonged duration of use, drug interactions and polypharmacy. The latter may be particularly problematic in the elderly. Monitoring systems, a partnership between patients, physicians and pharmacists and the provision of education and information to all concerned on safe self medication, are proposed strategies for maximising benefit and minimising risk. PMID- 11735660 TI - Comparative tolerability of first-generation selective estrogen receptor modulators in breast cancer treatment and prevention. AB - In general, the selective estrogen receptor modulators (SERMs) currently indicated for the treatment and prevention of breast cancer, i.e. tamoxifen and toremifene, are fairly well tolerated. However, tamoxifen has been shown to induce hepatocellular carcinomas in rats, but not in humans, and can increase the risk of endometrial cancer in humans by two to three times. Other potentially serious adverse effects which have been associated with tamoxifen and toremifene therapy include vasomotor symptoms, an increased risk of venous thromboembolic events, and an increased incidence of cataracts and ocular toxicity, fatty liver, and nonmalignant hepatic and uterine changes. In addition, long term tamoxifen use almost always results in resistance to the drug and, indeed, has actually been shown to promote tumour proliferation in human breast cancer cells. Both tamoxifen and toremifene display drug interactions with a variety of drug classes. The adverse events associated with these compounds have raised significant concerns regarding their widespread use for the treatment and prevention of breast cancer. In addition, because of the weakness and scarcity of the data on toremifene, any conclusions about its tolerability remain tentative until outcomes of ongoing clinical trials in the adjuvant setting are known. A third SERM, raloxifene, is the focus of several large randomised trials examining its efficacy in the prevention of breast cancer. At present, each potential adverse event needs to be weighed against potential benefits in the decision to undergo SERM treatment. An array of therapies is currently available for patients with breast cancer and women at increased risk of disease; the risk-to-benefit ratio for each agent should be carefully examined in determining the most advantageous regimen. PMID- 11735661 TI - Antimalarial drugs in systemic lupus erythematosus: use in pregnancy. AB - The 4-aminoquinoline radical containing antimalarial drugs are also used in the management of various connective tissue diseases including systemic lupus erythematosus (SLE) and rheumatoid arthritis. These agents are particularly useful for the management of inflammatory polyarthritis and skin disease. By raising the pH in intracellular compartments, these drugs interfere with normal phagocytic function which consequently enables them to interfere with antigen processing. Other actions include inhibition of platelet aggregation, this is advantageous in patients with phospholipid antibodies (aPL) which are known to predispose patients to recurrent arterial and venous clinical thrombotic events. Hydroxychloroquine has also been demonstrated to reduce serum lipid levels including cholesterol, triglycerides and low density lipoproteins. As it is now known that patients with SLE are at risk for accelerated artherogenesis and premature heart disease, this action may be an added benefit for these patients. The use of the 4-aminoquinoline radical containing antimalarial drugs during pregnancy is controversial. It is known that these agents can cross the placenta and are deposited in fetal pigmented tissues. These findings have led to the recommendation that these agents should be discontinued in pregnancy for patients with connective tissue diseases even though they have long been recommended for malarial prophylaxis in pregnant women travelling to malarial infested areas. Flares of SLE disease have been documented when these agents are discontinued and as flares of SLE disease activity are known to be detrimental to pregnancy outcome in patients with SLE, it is our opinion that these drugs should not be discontinued during pregnancy in a patient with lupus, particularly when the known terminal elimination half life is 1 to 2 months. PMID- 11735662 TI - Asthma medications and their potential adverse effects in the elderly: recommendations for prescribing. AB - The incidence of drug-induced adverse effects is likely to increase as a result of advanced age and exposure of elderly patients to polypharmacy. Therefore, pharmacological therapy of asthma and chronic obstructive pulmonary disease (COPD) in the elderly patient can be potentially hazardous. beta(2)-agonists, administered as therapy for asthma and COPD, have recognised systemic sequelae, such as hypokalaemia and chronotropic effects, which may be life-threatening in susceptible patients. Adverse effects such as hypokalaemia can be aggravated by concomitant treatment with other drugs promoting potassium loss including diuretics, corticosteroids and theophyllines. In addition, relatively minor adverse events associated with the administration of beta(2)-agonists, such as tremor and blood pressure changes, may be of significance to the elderly patient leading to impairment in the quality of life. However, long-term treatment with beta(2)-agonists may reduce the incidence of drug-induced adverse effects as a result of beta-receptor subsensitivity. Oral and inhaled corticosteroids have been used for the treatment of acute asthma and COPD in the elderly patient. Long term treatment with oral corticosteroids can result in serious systemic adverse effects such as suppressed adrenal function, bone loss, skin thinning and cataract formation. In contrast to beta(2)-agonists, oral corticosteroids can upregulate beta(2)-adrenoceptors and thereby potentiate the systemic sequelae of beta(2)-agonists. Hence, oral corticosteroids should be administered with caution for as short a duration as possible. Inhaled corticosteroids appear to be relatively well tolerated when administered at doses below approximately 1000 microg. However, larger doses of inhaled corticosteroids may affect hypothalamic pituitary-adrenal function and bone turnover. In the case of inhaled corticosteroids, spacer devices, often used in older patients who cannot operate metered dose inhalers, can potentiate the systemic sequelae of both corticosteroids and beta(2)-agonists. The use of theophyllines in the treatment of COPD or chronic asthma is controversial. Theophyllines have a wide adverse effect profile and are prone to drug-drug interactions. The adverse effects may be mild or life threatening and include nausea and vomiting or sinus and supraventricular tachycardias. Therefore, theophyllines should be prescribed with extreme caution to elderly patients with asthma or COPD. In contrast, inhaled anticholinergic drugs such as ipratropium bromide and oxitropium bromide are generally safe in elderly patients and have useful bronchodilator function. Commonly reported adverse effects are an unpleasant taste and dryness of the mouth. When used as first-line therapy, anticholinergic drugs may optimise the bronchodilator effects of low-dose inhaled beta(2)-agonists in patients with chronic airflow obstruction, and hence obviate the need for higher doses. PMID- 11735663 TI - Adverse drug reactions related to the use of NSAIDs with a focus on nimesulide: results of spontaneous reporting from a Northern Italian area. AB - OBJECTIVE: To analyse and compare the adverse drug reactions (ADRs) associated with the use of nimesulide with those associated with diclofenac, ketoprofen, and piroxicam, reported spontaneously in a northern Italian area (Veneto and Trentino). METHODS: Data were obtained from the spontaneous reporting system database of Veneto-Trentino, the principal contributor to the Italian spontaneous surveillance system. All case reports that occurred in association with all formulations of the nonsteroidal anti-inflammatory drugs (NSAIDs) under investigation during the period from January 1988 to December 2000, were analysed in detail. Sales data from June 1996 to May 1999 and prescription data, from 1997 to 2000 from the Veneto region were utilised to select the most widely used NSAIDs to be included in the study. The prescription data were also used to look at the drug use in relation to age. RESULTS: During the study period, 10 608 reports describing 16 571 adverse reactions were entered into the surveillance system. We found 207 case reports for nimesulide, 187 for diclofenac, 174 for ketoprofen, and 137 for piroxicam. Analysis of sales and prescription data revealed that in the Veneto region nimesulide was the most widely prescribed drug followed at a long distance by diclofenac, piroxicam and ketoprofen. No age related difference in the use of the four drugs was found. Analysis of the case reports revealed significantly different toxicity profiles for the four drugs. In particular, nimesulide was associated with fewer and less severe gastrointestinal (GI) ADRs compared with the other NSAIDs. Nimesulide was associated with about half the number of GI reactions (10.4%) than the other three NSAIDs (21.2% for diclofenac, 21.7% for ketoprofen, 18.6% for piroxicam). Two previously unreported reactions were also found for piroxicam and ketoprofen. CONCLUSIONS: Nimesulide is the most frequently used NSAID in Italy. Spontaneous reporting data suggest that nimesulide has the most favourable GI tolerability profile of the NSAIDs investigated, with few reports of severe GI reactions. A few reports of hepatic and renal impairment associated with nimesulide suggest caution in patients at risk. Age-related reporting analysis suggests a higher toxicity for diclofenac and piroxicam in the elderly compared with nimesulide and ketoprofen. This analysis of the Veneto-Trentino database on spontaneous reporting confirms that NSAIDs differ in their tolerability profile, and this fact should be taken into account in the choice of drugs in relation to patient characteristics. PMID- 11735664 TI - Acute renal failure in children: aetiology and management. AB - This review evaluates the various causes and management of acute renal failure (ARF) in children. ARF is defined as an abrupt decline in the renal regulation of water, electrolytes and acid-base balance, and continues to be an important factor contributing to the morbidity and mortality of critically ill infants and children. The common causes of ARF in children include acute tubular necrosis secondary to various causes (including congestive heart failure and sepsis), haemolytic uremic syndrome, and glomerulonephritis and urinary tract obstruction. Ischaemia, toxins (including drugs) as well as primary parenchymal disease, have to be considered and ARF can also be a complication of systemic disease. The basic principles of management are avoidance of life-threatening complications, maintenance of fluid and electrolyte balance, and nutritional support. Only a few patients require specific management of the underlying disorder, although it is important to diagnose these conditions. Knowledge about the use of drugs for the prevention of ARF is scarce. Mannitol, low-dose dopamine, calcium channel antagonists, atrial natriuretic peptide and albumin have been evaluated and, where possible, meta-analyses are cited. Mannitol treatment appears to be warranted prophylactically after paediatric renal transplantation. Albumin infusion can reverse prerenal ARF in children with nephritic syndrome. For treatment of the complications of hyperkalaemia and volume overload, salbutamol, insulin and glucose infusion and diuretics such as furosemide and sodium bicarbonate, are discussed. All of the major dialysis modalities (peritoneal dialysis, haemodialysis and continuous haemofiltration) can be used to provide equivalent solute clearance and ultrafiltration. The indication for, and the choice of the modality depend on the patient requirements and on local resources, and should involve the care of a paediatric nephrologist. Peritoneal dialysis requires minimal equipment and infrastructure, is easy to perform and remains the favoured modality of renal replacement therapy in children. However, continuous haemofiltration is an excellent alternative to peritoneal dialysis in patients with ARF and severe fluid overload. Dialysis remains the most important tool to bridge the time needed for recovery of renal function. There is increasing evidence that more intense use of dialysis may improve the overall prognosis. PMID- 11735665 TI - Prevention and management of infection in children with sickle cell anaemia. AB - Sickle cell anaemia (SCA) predisposes a child to infections for various reasons, including increased bone marrow turnover, poor perfusion and functional asplenia leading to decreased opsonisation of polysaccharide encapsulated organisms. Bacteria and viruses that most frequently cause serious infections in children with sickle cell disease are Streptococcus pneumoniae, Haemophilus influenzae type b, Salmonella spp., Escherichia coli, Staphylococcus aureus, Mycoplasma pneumoniae, Chlamydia pneumoniae, parvovirus B19 and hepatitis A, B and C viruses. Penicillin prophylaxis has decreased the incidence of infection-related morbidity and mortality significantly in children with SCA. Children <3 years of age are administered oral penicillin 125mg twice daily, and the dose is increased to 250mg twice daily for the >3 to 5 year age group. Adherence to the penicillin prophylactic regimen is recommended for children with SCA who are >5 years of age. For children with SCA who have recurrent invasive pneumococcal infections, an effort is made to keep the child on penicillin prophylaxis indefinitely. The administration of various childhood vaccines has also made an appreciable impact on the overall morbidity and mortality associated with infection in children with SCA. The administration of the heptavalent conjugate pneumococcal vaccine (PCV7) has provided control of invasive pneumococcal infections, and the prophylactic use of the H. influenzae type b conjugate vaccine has reduced the incidence of septicaemia and meningitis caused by this organism. Other vaccines used prophylactically in children with SCA include hepatitis A and B, and vaccines against influenza and varicella viruses. The immediate administration of intravenous antibacterials, after appropriate blood and urine cultures, is of great importance in the treatment of the febrile child with SCA. Ceftriaxone and cefotaxime have been recommended for the treatment of septic episodes in SCA associated with S. pneumoniae, Haemophilus and Salmonella spp. Infection with Yersinia enterocolitica may be treated with cefotaxime or an aminoglycoside. The prevalence of Helicobacter pylori infection in SCA is unknown. Effective therapies include metronidazole, tetracycline or amoxicillin. Parvovirus infections require supportive care and specific antiviral therapy is not indicated. The judicious use of antimicrobials is encouraged in view of the worldwide emergence of multidrug-resistant strains. The long term sequelae associated with infections in children with SCA can be decreased with the implementation of immunisation programmes and effective and prompt treatment with appropriate antibacterials. PMID- 11735666 TI - Effects of antibacterials on the unborn child: what is known and how should this influence prescribing. AB - Antibacterials are among the most commonly prescribed drugs worldwide. In general, infections occur in pregnant women at much the same rate as in the general population. However, as a result of physiological changes brought about by pregnancy, some infections, such as those of the urinary tract, may have an increased incidence. It is important to remember that almost every drug crosses the placenta, ensuring that the unborn fetus is also exposed. When prescribing an antibacterial agent to a pregnant woman, it is important that the mother is treated appropriately while at the same time protecting the unborn child. Certain factors need to be addressed, such as the possible teratogenic risk, changes in pharmacokinetics and the potential toxicity of the drug. In this paper we have reviewed various classes of antibacterials which are commonly used during pregnancy, including penicillins, beta-lactam inhibitors, cephalosporins, macrolides, aminoglycosides, tetracyclines, lincosamides, fluoroquinolones, sulfonamides, nitrofurans, and anti-tubercular agents. Some of these drugs have been on the market for many years, whereas others are relatively new and increasingly popular, despite the fact that the older drugs remain very effective. After reviewing the evidence-based information from epidemiological studies, it appears that most antibacterial agents can be used relatively safely during pregnancy. Women who are pregnant should not be denied appropriate antibacterial therapy because of a lack of information. It is possible to treat the mother, while protecting the unborn child, by prescribing an agent that the causative bacteria is sensitive to, rather than a perceived 'safer' option that may not effectively treat the infection and which may also add to the growing problem of bacterial resistance. PMID- 11735668 TI - Formulary restriction of selective serotonin reuptake inhibitors for depression: potential pitfalls. AB - The American healthcare market is currently estimated at more than 900 billion US dollars with double digit rising costs per year. Psychotropic agent costs have more than kept pace with market increases. Medication acquisition costs are an obvious focus for limiting costs in various care systems. Restrictive formularies are a common method of attempting to limit costs. To support our opinion that a single agent is ill advised, we explored the available evidence on the intended and unintended consequences of having a single or exclusive selective serotonin reuptake inhibitor (SSRI) on a formulary. Central to this position is an assumption of the interchangeability of SSRIs; we examined the evidence for and against this through a model to determine the probability of interchangeability. We conclude that the practice of having a single SSRI on the formulary for a healthcare plan seems ill founded. Patients who switch antidepressants remain in treatment 50% longer and cost approximately 50% more to treat in a more costly treatment setting. Giving the primary care physician several antidepressant choices can provide more options to continue treatment of his or her patient in the less expensive primary care setting. In terms of cost containment, formulary restrictions are far more likely to have the opposite effect. PMID- 11735669 TI - Which antifungal agent for onychomycosis? A pharmacoeconomic analysis. AB - The incidence of fungal nail infections is increasing and this is possibly because of several factors: better methods of detection, a growing population of immunocompromised patients who have a greater susceptibility to such infections, the increased use of immunosuppressive drugs, the increasing number of elderly people, worldwide travel, and the use of communal bathing facilities. Onychomycosis is a fungal infection of the fingernails and toenails that accounts for about 30% of all superficial fungal infections. It is characterised by nail discoloration, thickening and ultimately destruction of the nail plate. Management of this disease has improved significantly and treatment patterns have dramatically changed in recent years as a result of advances in new treatment options (e.g. oral antifungal agents) and changes in treatment regimens (e.g. pulse therapy). Also, newer drugs for onychomycosis have improved tolerability profiles compared with older agents. The overall costs of treating onychomycosis are substantial, and it has been estimated that direct cost for US Medicare patients with the disease is 43 million US dollars per year (year of costing not available). Pharmacoeconomic studies help in the decision-making process when selecting the most cost-effective antifungal agents to treat onychomycosis. To date there have been a number of national and international economic studies aimed at effectively assessing the efficacy and costs of the treatment options available to cure onychomycosis. The objectives of this paper are to (i) review the published findings regarding the epidemiology of onychomycosis; (ii) summarise the original pharmacoeconomic studies that describe the economic impact of the disease; and (iii) address the impact of the disease on patients' health related quality of life. PMID- 11735670 TI - Barrett's oesophagus: a review of costs of the illness. AB - Barrett's oesophagus is a premalignant complication that occurs in approximately 10% of patients with gastro-oesophageal reflux disease (GORD). In patients with Barrett's oesophagus, the risk of adenocarcinoma of the oesophagus approaches 0.5% per patient-years. Therefore, practice guidelines have been developed that suggest screening patients with GORD, particularly those with long-standing symptoms and those aged > or =50 years for the presence of Barrett's metaplasia. These guidelines also suggest performing surveillance endoscopy for the development of dysplasia and/or cancer in patients found to harbour Barrett's oesophagus at initial screening with the frequency of subsequent endoscopies dictated by the presence and grade of dysplasia. In patients with high-grade dysplasia and/or early adenocarcinoma, oesophagectomy is curative. Given the important clinical and economic implications of GORD complicated by Barrett's oesophagus, we review the costs associated with screening, surveillance and treatment for this condition. Although the majority of physicians recommend and/or perform surveillance for dysplasia in the setting of Barrett's oesophagus, differences in endoscopic technique, surveillance intervals and cancer perception among practitioners influence total costs. In the US, it is estimated that a population-wide surveillance program could potentially result in a total cost of 289.9 million US dollars. The outpatient management of Barrett's oesophagus is estimated to cost 1241 US dollars per year with medication use alone accounting for over half of the total costs. Cost-effectiveness analyses have been performed to evaluate the economic impact and benefit of surveillance for dysplasia and/or cancer. Studies to date have utilised several outcome measures such as life-years gained, quality-adjusted life-years and cases of cancer detected. Therefore, the incremental cost-effectiveness ratios reported have varied greatly and are particularly sensitive to the prevalence of Barrett's oesophagus in patients with GORD and the incidence of adenocarcinoma. Further epidemiological and clinical studies are likely to further define the economic impact of Barrett's oesophagus as a complication of GORD. PMID- 11735667 TI - Risks and benefits of nonsteroidal anti-inflammatory drugs in children: a comparison with paracetamol. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) possess antipyretic, analgesic and anti-inflammatory effects. They are frequently used in children and have numerous therapeutic indications, the most common ones being fever, postoperative pain and inflammatory disorders, such as juvenile idiopathic arthritis (JIA) and Kawasaki disease. Their major mechanism of action is through inhibition of prostaglandin biosynthesis by blockade of cyclo-oxygenase (COX). The disposition of most NSAIDs has been mainly studied in infants > or = 2 years of age. Compared with adults, the volume of distribution and clearance of NSAIDs such as diclofenac, ibuprofen (infants aged between 3 months and 2.5 years), ketorolac and nimesulide were increased in children. The elimination half-life was similar in children to that in adults. These pharmacokinetic differences might be clinically significant with the need for higher loading and/or maintenance doses in children. Ibuprofen, acetylsalicylic acid (ASA) and acetaminophen are the most frequently used agents for fever reduction in children. Over the past 20 years, because of the association between ASA use and Reye's syndrome, most of the interest has been directed toward ibuprofen and acetaminophen. In view of its comparable antipyretic efficacy, but superior tolerability profile, acetaminophen, when used appropriately with age-adapted formulations, should remain the first-line therapy in the treatment of childhood fever. At the moment, there is no scientific evidence to recommend simultaneous use of these two antipyretic drugs. Most NSAIDs provide mild to moderate analgesia, with the exception of ketorolac which has a strong analgesic activity. The analgesic efficacy of ketorolac, ketoprofen, diclofenac and ibuprofen in the treatment of postoperative pain has been mainly studied following a single dose, in children of > or = 1 year of age undergoing minor surgeries. In this setting, when used either alone or in adjunct to caudal or epidural anaesthesia, they were associated with an opioid-sparing effect and were well tolerated. With the exception of ketorolac use in children undergoing tonsillectomy, where controversy exists regarding the risk of postoperative haemorrhage, NSAIDs have not been associated with an increased risk of perioperative bleeding. NSAIDs are the first-line therapy in JIA. They appear to be equally effective and tolerated, with the exception of ASA which is associated with more adverse effects. ASA has been used for many years in the treatment of Kawasaki disease and is part of the standard modality of treatment in combination with intravenous gammaglobulins. More recently, lung inflammation associated with cystic fibrosis (CF) has become a new target for NSAIDs. Despite promising preliminary results with ibuprofen, numerous questions need to be answered before this new strategy becomes part of the conventional treatment of patients with CF. In summary, NSAIDs are effective in reducing fever, alleviating pain and reducing inflammation in children, with a good tolerance profile. Pharmacokinetic studies are needed to characterise the disposition of NSAIDs in very young infants in order to use them rationally. To date, no studies have been published on the disposition, tolerability and efficacy of specific COX-2 inhibitors in children. Further clinical experience with these agents in adults is warranted before undergoing trials with specific COX-2 inhibitors in children. PMID- 11735671 TI - Health-related quality of life and healthcare utilisation in patients with Parkinson's disease: impact of motor fluctuations and dyskinesias. AB - Idiopathic Parkinson's disease (PD) is a common chronic progressive neuro degenerative disorder associated with the progressive loss of dopaminergic neurons in the substantia nigra. The natural course of the disease may lead to severe disability despite a variety of pharmacological and surgical treatment options. Levodopa is still the most effective symptomatic treatment for PD; however, long term use can cause a number of adverse effects including motor complications, nausea and vomiting, postural hypotension and changes in mental status. The onset of motor complications marks a crucial point in the management of PD. They may present as changes between akinetic and mobile phases (motor fluctuations) or as abnormal involuntary movements (dyskinesias). After levodopa treatment for 3 to 5 years, motor complications occur in approximately 50% of patients, and after 10 years in >80% of patients. Treatment options have recently expanded as new drugs have been licensed and surgical procedures refined. Patients with motor complications present a demanding task in disease management, and often multiple drugs and high dosages are necessary to achieve only suboptimal control, resulting in increased healthcare utilisation. Costs increase considerably in patients with motor fluctuations and dyskinesias compared with patients without these symptoms. In a French study, 6-month direct medical costs per patient increased from 1648 euros (EUR) to EUR3028 in patients without and with motor fluctuations, respectively. In a recent French study a significant difference in monthly direct medical costs was found in patients with and without dyskinesias (EUR560 vs 170). Unfortunately, no data are available on the effect of motor complications on indirect costs. Several studies have shown that health related quality of life (HR-QOL) is reduced when motor fluctuations occur. This may also be true of dyskinesias, but because of the limited number of studies a definite conclusion is not yet possible. Recently, surgical treatment options have been used to deal with advanced PD and late stage complications. Although their effect on motor complications and HR-QOL is well documented, they result in increased costs (total medical cost: EUR28920) compared with drug treatment alone and are increasingly restricted by healthcare providers. The purpose of this article is to review the available data from pharmacotherapeutic. surgical and economic studies on HR-QOL and healthcare expenditure in patients with PD, with a major focus on the impact of motor fluctuations and dyskinesias. PMID- 11735672 TI - Incremental cost-effectiveness analysis comparing rofecoxib with nonselective NSAIDs in osteoarthritis: Ontario Ministry of Health perspective. AB - BACKGROUND: Clinical trials have shown rofecoxib, a selective inhibitor of cyclo oxygenase-2, to be associated with fewer gastrointestinal complications than non selective nonsteroidal anti-inflammatory drugs (NSAIDs). OBJECTIVE: To evaluate the potential clinical and economic consequences of rofecoxib prescription in Ontario, Canada, for patients with osteoarthritis (OA) aged >65 years who did not respond to paracetamol (acetaminophen) therapy. DESIGN: Decision analytic modelling study. METHODS: A model was constructed to compare rofecoxib and nonselective NSAIDs with respect to their gastrointestinal complications in patients with OA. The model had a 1-year horizon and considered direct medical costs from the perspective of the Ontario Ministry of Health. Event rates were estimated from a pooled analysis of 8 phase IIb/Ill clinical trials. The number of perforations, ulcers and bleeds (PUBs) with each strategy was used as the primary measure of effectiveness. RESULTS: In the base-case scenario, the expected total cost per patient-day on nonselective NSAIDs was 1.60 Canadian dollars (Can dollars) versus 1.67 Can dollars on rofecoxib (1999 values). Rofecoxib was associated with 0.0109 fewer PUBs per patient per year. The incremental cost to avoid 1 additional PUB by substituting rofecoxib for nonselective NSAIDs was 2247 Can dollars. The rofecoxib strategy became dominant if a gastroprotective agent was prescribed to more than 27.5% of the patients receiving nonselective NSAIDs. CONCLUSION: For patients with OA aged >65 years in whom paracetamol therapy has failed, rofecoxib may represent a cost-effective alternative to nonselective NSAIDs. Increased costs for drug acquisition are offset, in part. by avoidance of gastrointestinal complications and reduced use of gastroprotective agents. Rofecoxib may offer increased benefit among patients at a higher risk of serious gastrointestinal events. PMID- 11735673 TI - A cost-cost study comparing etanercept with infliximab in rheumatoid arthritis. AB - OBJECTIVE: The objective of this study was to compare the total costs associated with the administration of two different tumour necrosis factor (TNF) strategies used in the treatment of rheumatoid arthritis (RA): etanercept, a soluble TNF receptor that can be administered at home by subcutaneous injection, versus infliximab, an antibody that requires an intravenous infusion in a hospital outpatient setting. DESIGN AND SETTING: The main analytical framework of the study was a cost-cost analysis comparing the total annual costs associated with the administration of etanercept and infliximab in adult RA patients. The perspective of the study was that of the Dutch society. An economic model was constructed to determine the costs of both treatments. The cost evaluation included direct medical costs, direct nonmedical costs and indirect costs. The base-case analysis compared monotherapy with etanercept versus a combination therapy with infliximab and methotrexate. Data for the economic model came from published literature, expert opinion and official price and tariff lists. All costs were in 1999 values. PATIENTS AND PARTICIPANTS: The analysis was performed for the adult RA population eligible for treatment with etanercept or infliximab in The Netherlands. MAIN OUTCOME MEASURES AND RESULTS: The analysis showed that the total annual drug costs per patient do not differ substantially between infliximab and etanercept, with costs of Netherland guilders (NLG)31,526 (12,610 US dollars) and NLG31,334 (12,534 US dollars), respectively. However, the other medical costs (i.e. excluding the costs of the two drugs themselves) are substantially higher for infliximab due to the additional costs associated with administration in an outpatient clinic and the use of methotrexate [NLG 12,621 (5048 US dollars) versus NLG269 (107 US dollars) for etanercept]. The impact of direct nonmedical costs (transportation) and indirect costs were negligible. Overall treatment with infliximab is more expensive than treatment with etanercept with total costs of NLG45 115 (18,046 US dollars) and NLG3I,621 (12,648 US dollars), respectively (42.7% increase). CONCLUSIONS: Based on the assumptions used in the model, we may conclude that the use of etanercept compares favourably with infliximab from a budgetary and health economic perspective: the total costs are substantially lower when the efficacy of etanercept is assumed to be at least equivalent to the efficacy of infliximab. PMID- 11735674 TI - Economic evaluations during early (phase II) drug development: a role for clinical trial simulations? AB - Faced with increasing demands on demonstrating cost effectiveness, pharmaceutical companies are required to conduct pharmacoeconomic evaluations throughout the drug development programme. At present, there is particular emphasis in the literature on burden-of-illness studies and on economic evaluations conducted alongside phase III clinical trials but not on those conducted during phase II clinical trials. This article describes modelling techniques, namely clinical trial simulations (CTS), which are gaining popularity in the clinical research community, but which might also prove to be beneficial during the conduct of these early pharmacoeconomic evaluations. The basic concepts and structure of CTS are described by using published examples of simulations of antipsychotic and anticancer drugs. With the use of an illustrative example of a hypothetical cholinesterase inhibitor for Alzheimer's disease, an integrated CTS-based pharmacoeconomic evaluation is presented. The results demonstrate how the modelling may be of value in 'go/no-go' decisions during the drug development programme. PMID- 11735676 TI - Cost effectiveness of treatment options in advanced breast cancer in the UK. AB - OBJECTIVE: To compare clinical and economic study data for docetaxel, paclitaxel and vinorelbine in the treatment of anthracycline-resistant advanced breast cancer. STUDY DESIGN AND METHODS: A Markov decision-analysis model to simulate the clinical course of a 'typical' patient with advanced breast cancer during salvage chemotherapy was updated with response rates and adverse effect rates from phase III clinical trial data for docetaxel, paclitaxel and vinorelbine. Costs were taken from UK national databases and hospitals. Utilities were estimated from 30 oncology nurses in the UK using the standard gamble method. PERSPECTIVE: National Health Service. RESULTS: When compared with other chemotherapeutic agents, docetaxel has been shown to increase response rate, time to progression and survival in patients with advanced breast cancer. In the base case analysis, the incremental cost-utility ratio for docetaxel versus paclitaxel was pound1995 per quality-adjusted life year (QALY) gained (1998 values). The incremental cost-utility ratio for docetaxel versus vinorelbine was pound14 055 per QALY gained. In the comparison with vinorelbine, docetaxel provided the equivalent of an additional 92 days of perfect health. Sensitivity analyses confirmed the robustness of the model and the validity of the base-case analysis results. Even in the worst case scenarios, docetaxel remained cost effective compared with paclitaxel and vinorelbine. CONCLUSIONS: These findings support the use of the taxoids, notably docetaxel, in the management of advanced breast cancer. PMID- 11735675 TI - Quality-of-life assessment in patients with benign prostatic hyperplasia: effects of various interventions. AB - A literature search was conducted to review quality-of-life (QOL) measurement in patients with benign prostatic hypertrophy (BPH). The different QOL instruments are discussed in relation to their correlation with symptom evaluation in studies of treatment options for BPH. Symptom evaluation of BPH has been standardised internationally with the International Prostatic Symptom Score (IPSS), but there is neither agreement nor data to decide which QOL instrument is preferable. The most widely used QOL instrument is the disease-specific QOL single question added to the IPSS. Other QOL instruments have been used, but none has gained unanimous approval. The results of QOL assessments obtained from comparative clinical studies of treatment options for BPH are discussed. These studies compare treatment options such as watchful waiting, drug treatment and surgery. Disease specific QOL domains (interference with daily activities) tend to improve more with treatment interventions than general health measures (i.e. general well being). The use of QOL instruments to evaluate patients with BPH, and their many treatment options, is still open to debate with regard to which instruments are preferred and their importance to the clinical evaluation of the patient. The challenge remains to find an acceptable disease-specific QOL instrument that adds information to currently used disease measures of BPH. PMID- 11735677 TI - Cost-effectiveness analysis and the consistency of decision making: evidence from pharmaceutical reimbursement in australia (1991 to 1996). AB - OBJECTIVE: The principle aim of this study was to generate a league table of drugs considered by the Australian Pharmaceutical Benefits Advisory Committee (PBAC) for reimbursement. The table was used to test the hypothesis that decisions made by the PBAC are consistent with the maxim of economic efficiency. In addition, we explored whether the past decisions by the PBAC revealed a threshold incremental cost-effectiveness ratio beyond which the PBAC is not prepared to recommend reimbursement of a drug. METHODS: All 355 submissions made to the PBAC between January 1991 and June 1996 were reviewed. Submissions using cost per life-year gained (26 submissions) or the cost per quality adjusted life year (QALY) gained (9 submissions) were ranked in a league table and compared with advice given by the PBAC about that drug. The confidentiality restrictions for the submissions require that the individual drug details cannot be revealed in this article. RESULTS: There was a statistically significant difference between the cost per life-year gained for drugs that were recommended for listing and those that were not, suggesting that the PBAC has been broadly consistent with the use of economic efficiency as a criterion for decision making. We did not find an explicit threshold beyond which the PBAC was unwilling to pay for additional life years gained. However, between 1992 and 1996 the PBAC appears to have been unlikely to recommend a drug for listing if the additional cost per life-year exceeded 76 000 Australian dollars [$AU] (1998/1999 values) and was unlikely to reject a drug for which the additional cost per life-year gained was less than $AU42 000. The cost-effectiveness ratio was not the only factor determining the reimbursement decision. CONCLUSIONS: The results of this preliminary study indicate that decisions to recommend a drug for listing by the PBAC in the last few years have, by and large, been consistent with the notion of economic efficiency. PMID- 11735678 TI - Paclitaxel: a pharmacoeconomic review of its use in non-small cell lung cancer. AB - A number of first-line chemotherapy options for patients with advanced non-small cell lung cancer (NSCLC) are advocated in treatment guidelines and/or by various clinical investigators. Platinum-based chemotherapy has clearly demonstrated efficacy in patients with advanced NSCLC and is generally recommended as first line therapy, although there is increasing interest in the use of non-platinum chemotherapy regimens. Among the platinum-based combinations currently used in clinical practice are regimens such as cisplatin or carboplatin combined with paclitaxel, vinorelbine, gemcitabine, docetaxel or irinotecan. The particular combinations employed may vary between institutions and geographical regions. Several pharmacoeconomic analyses have been conducted on paclitaxel in NSCLC and most have focused on its use in combination with cisplatin. In terms of clinical efficacy, paclitaxel-cisplatin combinations achieved significantly higher response rates than teniposide plus cisplatin or etoposide plus cisplatin (previously thought to be among the more effective regimens available) in two large randomised trials. One of these studies showed a survival advantage for paclitaxel plus cisplatin [with or without a granulocyte colony-stimulating factor (G-CSF)] compared with etoposide plus cisplatin. A Canadian cost effectiveness analysis incorporated data from one of the large randomised comparative trials and showed that the incremental cost per life-year saved for outpatient administration of paclitaxel plus cisplatin versus etoposide plus cisplatin was $US 22181 (30619 Canadian dollars; $Can) [1997 costs]. A European analysis incorporated data from the other large randomised study and showed slightly higher costs per responder for paclitaxel plus cisplatin than for teniposide plus cisplatin in The Netherlands ($US 30769 vs $US 29592) and Spain ($US 19 923 vs $US 19724) but lower costs per responder in Belgium ($US 22852 vs $US 25000) and France ($US28 080 vs $US 34747) [1995/96 costs]. In other cost effectiveness analyses, paclitaxel plus cisplatin was associated with a cost per life-year saved relative to best supportive care of approximately $US 10000 in a US study (year of costing not reported) or $US 11200 in a Canadian analysis ($Can 15400; 1995 costs). Results were less favourable when combining paclitaxel with carboplatin instead of cisplatin and particularly when G-CSF was added to paclitaxel plus cisplatin. The Canadian study incorporated the concept of extended dominance in a threshold analysis and ranked paclitaxel plus cisplatin first among several comparator regimens (including vinorelbine plus cisplatin) when the threshold level was $Can 75000 ($US 54526) per life-year saved or per quality-adjusted life-year gained (1995 values). CONCLUSION: Current treatment guidelines for advanced NSCLC recognise paclitaxel-platinum combinations as one of the first-line chemotherapy treatment options. In two large head-to-head comparative clinical trials, paclitaxel plus cisplatin was associated with significantly greater response rates than cisplatin in combination with either teniposide or etoposide, and a survival advantage was shown for paclitaxel plus cisplatin (with or without G-CSF) over etoposide plus cisplatin. There are limitations to the currently available pharmacoeconomic data and further economic analyses of paclitaxel-carboplatin regimens are warranted, as this combination is widely used in NSCLC and appears to have some clinical advantages over paclitaxel plus cisplatin in terms of ease of administration and tolerability profile. Nevertheless, results of various cost-effectiveness studies support the use of paclitaxel-platinum combinations, particularly paclitaxel plus cisplatin, as a first-line chemotherapy treatment option in patients with advanced NSCLC. PMID- 11735679 TI - Piperacillin/tazobactam: a pharmacoeconomic review of its use in moderate to severe bacterial infections. AB - Piperacillin/tazobactam is a beta-lactam/beta-lactamase inhibitor combination with a broad spectrum of antibacterial activity against most Gram-positive and Gram-negative aerobic bacteria and anaerobic bacteria. Piperacillin/tazobactam is effective and well-tolerated in patients with lower respiratory tract infections (LRTI), intra-abdominal infections, skin and soft tissue infections, and febrile neutropenia. In comparative clinical trials against various other antibacterial regimens, piperacillin/tazobactam has shown higher clinical success rates, particularly in the treatment of patients with intra-abdominal infections and febrile neutropenia. Cost analyses of piperacillin/tazobactam have been variable, in part, because of differences in specific costs included. Three US cost analyses found that piperacillin/tazobactam had lower total medical costs than clindamycin plus gentamicin or imipenem/cilastatin in intra-abdominal infections, and ticarcillin/ clavulanic acid in community-acquired pneumonia. Piperacillin/tazobactam plus amikacin had lower total costs than ceftazidime plus amikacin in another cost analysis of patients with febrile neutropenic episodes modelled in nine European countries. However, piperacillin/tazobactam plus tobramycin was more costly than ceftazidime plus tobramycin in hospital-acquired pneumonia in a US cost analysis. In cost-effectiveness analyses, all studies of intra-abdominal infections, pneumonia and febrile neutropenic episodes consistently reported lower costs per unit of effectiveness versus comparators. Piperacillin/tazobactam was dominant (greater efficacy and lower costs) versus imipenem/cilastatin in intra-abdominal infections and ceftriaxone, ciprofloxacin or meropenem in pneumonia. Piperacillin/tazobactam plus amikacin was dominant over ceftazidime plus amikacin in the treatment of febrile neutropenic episodes. In a cost-effectiveness analysis of skin and soft tissue infection, piperacillin/tazobactam had lower costs per successfully treated patient than ceftriaxone or cefotaxime, but a slightly higher cost-effectiveness ratio than amoxicillin/clavulanic acid. All cost-effectiveness analyses were based on decision-analytical models. CONCLUSIONS: Piperacillin/tazobactam is likely to reduce overall treatment costs of moderate to severe bacterial infections by increasing initial treatment success, thereby reducing the length of hospital stay and the use of additional antibacterials. Piperacillin/tazobactam has shown clinical and economic advantages over standard antibacterial regimens in the treatment of intra-abdominal infections, LRTIs, febrile episodes in patients with neutropenia, and skin and soft tissue infections, although more complete published data are needed to confirm these results. Present data regarding clinical efficacy, bacterial resistance and costs would support the use of piperacillin/tazobactam as an empirical first-line option in moderate to severe bacterial infections. PMID- 11735680 TI - Prescription of resistance training for healthy populations. AB - Although there are well documented protective health benefits conferred by regular physical activity, most individuals of all ages are not physically active at a level for sufficient maintenance of health. Consequently, a major public health goal is to improve the collective health and fitness levels of all individuals. The American College of Sports Medicine (ACSM) and other international organisations have established guidelines for comprehensive exercise programmes composed of aerobic, flexibility and resistance-exercise training. Resistance training is the most effective method available for maintaining and increasing lean body mass and improving muscular strength and endurance. Furthermore, there is an increasing amount of evidence suggesting that resistance training may significantly improve many health factors associated with the prevention of chronic diseases. These health benefits can be safely obtained by most segments of the population when prescribed appropriate resistance exercise programmes. Resistance-training programmes should be tailored to meet the needs and goals of the individual and should incorporate a variety of exercises performed at a sufficient intensity to enhance the development and maintenance of muscular strength and endurance, and lean body mass. A minimum of 1 set of 8 to 10 exercises (multi-joint and single joint) that involve the major muscle groups should be performed 2 to 3 times a week for healthy participants of all ages. More technical and advanced training including periodised multiple set regimens and/or advanced exercises may be more appropriate for individuals whose goals include maximum gains in strength and lean body mass. However, the existing literature supports the guidelines as outlined in this paper for children and adults of all ages seeking the health and fitness benefits associated with resistance training. PMID- 11735681 TI - Brain microdialysis in exercise research. AB - During the last 5 to 10 years, the microdialysis technique has been used to explore neurotransmitter release during exercise. Microdialysis can collect virtually any substance from the brains of freely moving animals with a limited amount of tissue trauma. It allows the measurement of local neurotransmitter release in combination with ongoing behavioural changes such as exercise. Several groups examined the effect of treadmill running on extracellular neurotransmitter levels. Microdialysis probes were implanted in different brain areas to monitor diverse aspects of locomotion (striatum, hippocampus, nucleus accumbens, frontal cortex, spinal cord), food reward (hypothalamus, hippocampus, cerebral cortex), thermoregulation (hypothalamus). Some studies combined microdialysis with running on a treadmill to evaluate motor deficit and improvement following dopaminergic grafts in 6-hydroxydopamine lesioned rats, or combined proton nuclear magnetic resonance spectroscopy and cortical microdialysis to observe intra- plus extracellular brain glucose variations. This method allows us to understand neurotransmitter systems underlying normal physiological function and behaviour. Because of the growing interest in exercise and brain functioning, it should be possible to investigate increasingly subtle behavioural and physiological changes within the central nervous system. There is now compelling evidence that regular physical activity is associated with significant physiological, psychological and social benefits in the general population. In contrast with our knowledge about the peripheral adaptations to exercise, studies relating exercise to brain neurotransmitter levels are scarce. It is of interest to examine the effect of short and long term exercise on neurotransmitter release, since movement initiation and control of locomotion have been shown to be related to striatal neurotransmitter function, and one of the possible therapeutic modalities in movement, and mental disorders is exercise therapy. Until very recently most experimental studies on brain chemistry were conducted with postmortem tissue. However, in part because of shortcomings with postmortem methods, and in part because of the desire to be able to directly relate neurochemistry to behaviour, there has been considerable interest in the development of 'in vivo' neurochemical methods. Because total tissue levels may easily mask small but important neurochemical changes related to activity, it is important to sample directly in the extracellular compartment of nervous tissue in living animals. Since the chemical interplay between cells occurs in the extracellular fluid, there was a need to access this compartment in the intact brain of living and freely moving animals. Estimation of the transmitter content in this compartment is believed to be directly related to the concentration at the site where these compounds are functionally released: in the synaptic cleft. As measurements in the synapse are not yet possible, in vivo measurements in the extracellular fluid appear to provide the most directly relevant information currently available. This article provides an overview of the in vivo microdialysis technique as a method for measuring in the extracellular space, and its application in exercise science. Although this technique has been used in different tissues such as brain, adipose tissue, spinal cord and muscle, in animals as well as humans, we will focus on the use of this in vivo method in brain tissue. Recently two excellent reviews on the application of microdialysis in human experiments especially in subcutaneous tissue have been published, and we refer the interested reader to these articles. PMID- 11735682 TI - Is it possible to prevent sports injuries? Review of controlled clinical trials and recommendations for future work. AB - Sports injuries are one of the most common injuries in modern western societies. Treating sports injuries is often difficult, expensive and time consuming, and thus, preventive strategies and activities are justified on medical as well as economic grounds. A successful injury surveillance and prevention requires valid pre- and post-intervention data on the extent of the problem. The aetiology, risk factors and exact mechanisms of injuries need to be identified before initiating a measure or programme for preventing sports injuries, and measurement of the outcome (injury) must include a standardised definition of the injury and its severity, as well as a systematic method of collecting the information. Valid and reliable measurement of the exposure includes exact information about the population at risk and exposure time. The true efficacy of a preventive measure or programme can be best evaluated through a well-planned randomised trial. Until now, 16 randomised, controlled trials (RCT) have been published on prevention of sports injuries. According to these RCT, the general injury rate can be reduced by a multifactorial injury prevention programme in soccer (relative risk 0.25, p < 0.001, in the intervention group), or by ankle disk training, combined with a thorough warm-up, in European team handball [odds ratio 0.17; 95% confidence interval (CI) 0.09 to 0.32, p < 0.01]. Ankle sprains can be prevented by ankle supports (i.e. semirigid orthoses or air-cast braces) in high-risk sporting activities, such as soccer and basketball (Peto odds ratio 0.49; 95% CI 0.37 to 0.66), and stress fractures of the lower limb by the use of shock-absorbing insoles in footwear (Peto odds ratio 0.47; 95% CI 0.30 to 0.76). In future studies, it is extremely important for researches to seek consultation with epidemiologists and statisticians to be certain that the study hypothesis is appropriate and that the methodology can lead to reliable and valid information. Further well-designed randomised studies are needed on preventive actions and devices that are in common use, such as preseason medical screenings, warming up, proprioceptive training, stretching, muscle strengthening, taping, protective equipment, rehabilitation programmes and education interventions (such as increasing general injury awareness among a team). The effect of a planned rule change on the injury risk in a particular sport could be tested via a RCT before execution of the change. The most urgent needs are in commonly practised or high risk sports, such as soccer, American football, rugby, ice hockey, European team handball, karate, floorball, basketball, downhill skiing and motor sports. PMID- 11735683 TI - Magnetic resonance imaging of the knee. AB - Magnetic resonance imaging (MRI) has revolutionised diagnostic imaging of the knee. It has evolved significantly since Kean described healthy and pathologic knee anatomy in 1983. This innovative technology allows superior soft-tissue detail with multiplanar imaging capability that provides accurate evaluation of intra- and extra-articular structures of the knee not demonstrated with other imaging modalities. The development and advancements in MRI and the introduction of high-resolution coils have provided a noninvasive, nonoperator dependent, cost effective means to diagnose knee pathology. MRI is well tolerated by patients, widely accepted by evaluating physicians, and assists in distinguishing pathologic knee conditions that may have similar clinical signs and symptoms (i.e. meniscal tears, osteochondral lesions). This paper presents an overview of MRI of the knee and focuses on the MRI findings in a number of common pathologic conditions. PMID- 11735684 TI - Estrogen replacement therapy and female athletes: current issues. AB - Physicians commonly recommend estrogen replacement as treatment for exercise associated amenorrhoea. While the evidence shows that the basis of the amenorrhoea is estrogen deficiency, it is not clear that it is the only factor in the development of lowered bone density found in oligo-amenorrhoeic female athletes. Nutritional factors, significant in the development of the reproductive dysfunction, could also contribute to bone loss. No randomised, controlled studies of estrogen replacement in athletes have been published. However, one nonrandomised study of a small group of athletes does suggest that there are significant gains in bone density to be made by the initiation of estrogen therapy. More research is clearly needed. PMID- 11735687 TI - The legacy of Harvey Cushing. PMID- 11735685 TI - Blood lipid and lipoprotein adaptations to exercise: a quantitative analysis. AB - Dose-response relationships between exercise training volume and blood lipid changes suggest that exercise can favourably alter blood lipids at low training volumes, although the effects may not be observable until certain exercise thresholds are met. The thresholds established from cross-sectional literature occur at training volumes of 24 to 32 km (15 to 20 miles) per week of brisk walking or jogging and elicit between 1200 to 2200 kcal/wk. This range of weekly energy expenditure is associated with 2 to 3 mg/dl increases in high-density lipoprotein-cholesterol (HDL-C) and triglyceride (TG) reductions of 8 to 20 mg/dl. Evidence from cross-sectional studies indicates that greater changes in HDL-C levels can be expected with additional increases in exercise training volume. HDL-C and TG changes are often observed after training regimens requiring energy expenditures similar to those characterised from cross-sectional data. Training programmes that elicit 1200 to 2200 kcal/wk in exercise are often effective at elevating HDL-C levels from 2 to 8 mg/dl, and lowering TG levels by 5 to 38 mg/dl. Exercise training seldom alters total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C). However, this range of weekly exercise energy expenditure is also associated with TC and LDL-C reductions when they are reported. The frequency and extent to which most of these lipid changes are reported are similar in both genders, with the exception of TG. Thus, for most individuals, the positive effects of regular exercise are exerted on blood lipids at low training volumes and accrue so that noticeable differences frequently occur with weekly energy expenditures of 1200 to 2200 kcal/wk. It appears that weekly exercise caloric expenditures that meet or exceed the higher end of this range are more likely to produce the desired lipid changes. This amount of physical activity, performed at moderate intensities, is reasonable and attainable for most individuals and is within the American College of Sports Medicine's currently recommended range for healthy adults. PMID- 11735688 TI - Viral load test reports: a description of content from a sample of US laboratories. AB - CONTEXT: Human immunodeficiency virus (HIV) RNA testing (viral load testing) is increasingly important in the care of patients infected with HIV-1 to determine when to initiate, monitor, and change antiretroviral therapy. Patient viral load testing information is communicated to the clinician through the laboratory test report. OBJECTIVES: To examine the format and information used in reporting viral load testing results and determine the clarity of the information provided in these reports. DESIGN: Patient test reports with all personal identifiers removed were requested of viral load testing laboratories participating in a telephone survey of laboratory practices. Hospital, independent, health department, and "other type" laboratories identified as university-associated laboratories participated in the telephone survey. RESULTS: Thirty-seven unique test reports were collected. All laboratories reported results in copies/mL, while 14% also reported results as "log(10) copies/mL." The test kit was identified by only 24% of the laboratories. Reportable ranges were specified by 70% of the laboratories, but there was considerable variation in terminology. One laboratory reported a viral load copy number below the manufacturer's test kit lower limit of sensitivity. The layout and format differed among reports. Some results were expressed in log(10), others contained nonsignificant integers, while others contained exponential numbers. Supplemental information in some reports included previous patient test results and significance of changes from baseline. The format of some reports made it difficult to read the report information and interpret the testing results. CONCLUSION: This study emphasizes the importance of standardizing the reporting of HIV-1 viral load test results to minimize result misinterpretation and incorrect treatment. PMID- 11735689 TI - Aortic and mitral fenfluramine-phentermine valvulopathy in 64 patients treated with anorectic agents. AB - CONTEXT: Few published studies of the pathology of fenfluramine-phentermine (fen phen) valvulopathy, described by Connolly and colleagues in 1997, have appeared. OBJECTIVES: To define temporal changes in the morphology of the stuck-on plaques and to determine whether the plaques progress or regress after cessation of fen phen. METHODS: The available clinical information and pathology material from 35 aortic valves (AVs) and 43 mitral valves (MVs) from 64 patients were reviewed. RESULTS: The valves fell into 3 groups: 17 AVs and 28 MVs had fen-phen lesions only, 2 AVs and 7 MVs had fen-phen changes associated with other valve diseases, and 16 AVs and 8 MVs had no fen-phen changes. Fenfluramine-phentermine attributable dysfunction was regurgitation in all instances. Typical plaques showed proliferation of myofibroblastic cells with myxoid stroma. Small vascular channels and slight lymphocytic accumulations were often present. Deeper parts of some plaques had dense fibroelastic tissue underlying typical plaque. CONCLUSIONS: Considerable individual variation in the time course of anorectic agent use and the severity of fen-phen valvulopathy was observed. Possible plaque regression could not be assessed from this study. The observations suggest that in some patients fen-phen-induced plaques may continue to have surface proliferation despite drug withdrawal. PMID- 11735686 TI - Long-term metabolic and skeletal muscle adaptations to short-sprint training: implications for sprint training and tapering. AB - The adaptations of muscle to sprint training can be separated into metabolic and morphological changes. Enzyme adaptations represent a major metabolic adaptation to sprint training, with the enzymes of all three energy systems showing signs of adaptation to training and some evidence of a return to baseline levels with detraining. Myokinase and creatine phosphokinase have shown small increases as a result of short-sprint training in some studies and elite sprinters appear better able to rapidly breakdown phosphocreatine (PCr) than the sub-elite. No changes in these enzyme levels have been reported as a result of detraining. Similarly, glycolytic enzyme activity (notably lactate dehydrogenase, phosphofructokinase and glycogen phosphorylase) has been shown to increase after training consisting of either long (>10-second) or short (<10-second) sprints. Evidence suggests that these enzymes return to pre-training levels after somewhere between 7 weeks and 6 months of detraining. Mitochondrial enzyme activity also increases after sprint training, particularly when long sprints or short recovery between short sprints are used as the training stimulus. Morphological adaptations to sprint training include changes in muscle fibre type, sarcoplasmic reticulum, and fibre cross sectional area. An appropriate sprint training programme could be expected to induce a shift toward type IIa muscle, increase muscle cross-sectional area and increase the sarcoplasmic reticulum volume to aid release of Ca(2+). Training volume and/or frequency of sprint training in excess of what is optimal for an individual, however, will induce a shift toward slower muscle contractile characteristics. In contrast, detraining appears to shift the contractile characteristics towards type IIb, although muscle atrophy is also likely to occur. Muscle conduction velocity appears to be a potential non-invasive method of monitoring contractile changes in response to sprint training and detraining. In summary, adaptation to sprint training is clearly dependent on the duration of sprinting, recovery between repetitions, total volume and frequency of training bouts. These variables have profound effects on the metabolic, structural and performance adaptations from a sprint-training programme and these changes take a considerable period of time to return to baseline after a period of detraining. However, the complexity of the interaction between the aforementioned variables and training adaptation combined with individual differences is clearly disruptive to the transfer of knowledge and advice from laboratory to coach to athlete. PMID- 11735690 TI - Metastatic angiosarcoma masquerading as diffuse pulmonary hemorrhage: clinicopathologic analysis of 7 new patients. AB - CONTEXT: Diffuse pulmonary hemorrhage is an uncommon presenting manifestation of angiosarcoma. OBJECTIVE: To review the clinical, radiologic, and pathologic findings of patients with metastatic angiosarcoma who presented with diffuse pulmonary hemorrhage. DESIGN: Patients fulfilling inclusion criteria were identified from the consultation files. Clinical and radiologic data were obtained from referring pathologists. Histologic slides were reviewed in all patients. RESULTS: Our patients included 6 men and 1 woman, aged 31 to 73 years; 4 patients were younger than 40 years. Six patients presented with hemoptysis, and all had diffuse abnormalities on radiographic studies. Clinical considerations prior to biopsy included pulmonary hemorrhage syndrome (n = 2), acute respiratory failure (n = 1), and infection (n = 1). Metastatic disease was included in the differential diagnosis in only 1 patient. None had a diagnosis of malignancy prior to lung biopsy. All biopsies showed hemorrhage associated with atypical epithelioid and spindle cells, forming anastomosing vascular channels, distributed along and within lymphatics and arteries. Neoplastic cells were immunoreactive for factor VIII-related protein or CD31 in all cases for which special stains were available. Three patients with complete follow-up died of their disease. Primary sites were discovered in all 3 patients who underwent autopsy examination. Two tumors arose in the heart and 1 in the pelvic soft tissues. One additional patient had a likely primary site identified in the right atrium by cardiac ultrasound and was subsequently lost to follow-up. CONCLUSION: Angiosarcoma should be included in the differential diagnosis of diffuse pulmonary hemorrhage, especially in young adults. PMID- 11735691 TI - Clear cell odontogenic carcinoma: a clinicopathologic and immunocytochemical study of 5 cases. AB - CONTEXT: Odontogenic tumor composed predominantly of clear cells is a rare neoplasm of the jaws that was initially designated as clear cell odontogenic tumor. Subsequent reports in the literature, however, all have indicated that this tumor exhibits an aggressive behavior characterized by infiltrative local growth, recurrence, or metastases. OBJECTIVE: To ascertain the clinicomorphologic features and biologic behavior of this tumor group, we present 5 additional cases using the term clear cell odontogenic carcinoma. DESIGN: Histologic and immunohistochemical examinations were performed on clear cell odontogenic carcinomas from 5 patients. Clinical and follow-up data were recorded, and the literature was reviewed. RESULTS: The patients were 1 man and 4 women with an average age of 42.4 years at diagnosis. Three tumors occurred in the mandible and the other 2 in the maxilla. Four of the 5 cases occurred in the anterior or premolar areas of the jaws, appearing as poorly marginated radiolucencies. All cases consisted of islands and sheets of large clear cells and small basaloid cells with scanty eosinophilic cytoplasm, which were separated by thin, mature, fibrous septae. The tumors were unencapsulated and showed aggressive infiltration of the surrounding muscle and perineural tissues. Most of the clear cells contained cytoplasmic glycogen. Immunocytochemically, the tumor cells were positive for pan-keratin, cytokeratin 19, and epithelial membrane antigen, but were negative for vimentin, S100 protein, desmin, smooth muscle actin, human melanoma antigen (HMB-45), and alpha(1)-antichymotrypsin. Follow-up data showed 4 of 5 patients to have multiple local recurrences, and 1 of these 4 had submandibular lymph node metastasis and a fatal outcome due to uncontrollable tumor growth. CONCLUSION: These results suggest that odontogenic clear cell neoplasms are at least low-grade malignancies and should be classified as carcinomas. PMID- 11735692 TI - Contribution of nonsteroidal anti-inflammatory drugs to deaths associated with peptic ulcer disease: a prospective toxicological analysis of autopsy blood samples. AB - CONTEXT: Of the side effects occurring in temporal association with the use of nonsteroidal anti-inflammatory drugs (NSAIDs), peptic ulcer disease is reported most often. OBJECTIVES: To (1) provide information on the temporal association between fatal peptic ulcer presenting as sudden death and NSAID use prior to death, and (2) to examine the diagnostic efficiency of postmortem determination of NSAID levels using high-pressure liquid chromatography. DESIGN: Prospective autopsy study of all cases of sudden death associated with peptic ulcer disease from a total of 1139 medicolegal autopsies performed during a 12-month period. METHODS: Postmortem femoral blood samples were analyzed for NSAIDs using high pressure liquid chromatography, and specimens of gastric and duodenal mucosa were examined for coexisting pathologic conditions. RESULTS: Twelve fatalities that occurred out of hospital as a result of peptic ulcer disease and presented as sudden death were identified. Autopsy blood samples were positive for NSAIDs in 7 cases (ibuprofen in 4 cases, levels 0.8 to 1.4 microg/mL; diclofenac in 2 cases, levels 0.6 and 1.6 microg/mL; and ketoprofen in 1 case, level 0.3 microg/mL). The ages of the affected individuals (3 men, 4 women) ranged from 43 to 60 years. No other drugs, including corticosteroids, anticoagulants, salicylic acid, and salicylates, were present. Microscopic examination revealed no pathologic antemortem mucosal conditions in any of the cases. CONCLUSIONS: For the postmortem elucidation of etiopathogenetic factors contributing to fatal peptic ulcer disease, high-pressure liquid chromatography to determine NSAID levels in autopsy blood samples is of considerable diagnostic benefit, especially when combined with histology. The number of cases of sudden death involving younger individuals dying as a result of peptic ulcer disease in temporal association with preceding use of NSAIDs seems to be underestimated from the clinical viewpoint due to the underrepresentation of out-of-hospital fatalities in the field of clinical pathology. PMID- 11735693 TI - Aspiration cytology of pediatric solitary papillary hyperplastic thyroid nodule. AB - CONTEXT: Solitary papillary hyperplastic thyroid nodules (SPHTNs) are frequently encountered in children and teenagers. Although the histologic features are well described, to the best of our knowledge, cytologic findings have not been reported. OBJECTIVES: To review the cytologic features of histologically proven SPHTNs and to identify the potential diagnostic pitfalls in cytologic diagnosis. MATERIALS AND METHODS: Fine-needle aspiration cytology of 3 histologically proven SPHTNs was reviewed. RESULTS: Two girls and 1 boy (ages 11, 12, and 15 years) were affected. The cytologic diagnosis in all 3 cases was suspicious for papillary thyroid carcinoma (PTC). The spectrum of cytologic findings included broad flat sheets and 3-dimensional clusters with fire flares. There was mild to moderate nuclear pleomorphism and nuclear atypia. Short nonbranching papillae with transgressing vessels shown to represent hyperplastic papillae on histologic sections were identified in all cases. The background contained giant cells, histiocytes, and watery and inspissated colloid. Although nuclear grooves were identified in occasional cells, intranuclear inclusions were absent. A cell block section (1 case) and histologic sections of SPHTNs (2 cases) were immunohistochemically negative for cytokeratin 19. CONCLUSIONS: Fine-needle aspiration of SPHTNs may be difficult to interpret accurately and can result in false-positive diagnosis of PTC. Although it shares several cytologic features with PTC, the presence of fire flares and short nonbranching papillae, as well as lack of intranuclear inclusions and watery and inspissated colloid in SPHTN appear to be useful features that are helpful in distinguishing SPHTN from PTC. Negative immunohistochemical staining for cytokeratin 19 is useful in excluding a diagnosis of PTC. PMID- 11735694 TI - Collagenous gastritis associated with lymphocytic gastritis and celiac disease. AB - Collagenous gastritis is a rare disorder, with only 8 cases reported in the literature, 2 in children and 6 in adults. We report an additional case of collagenous gastritis in a 42-year-old man with celiac disease. A thickened (>10 microm) subepithelial collagen band with entrapped capillaries, fibroblasts, and inflammatory cells was seen in the stomach, associated with lymphocytic gastritis. The duodenal mucosa showed severe villous atrophy but no subepithelial collagen deposition. No evidence of lymphocytic or collagenous colitis was found in the colon. The patient became symptom-free on a gluten exclusion diet and showed partial improvement of histopathologic findings after 3 months. Collagenous gastritis is a rare disease, but a wider recognition of its histopathologic features and clinical associations may bring more cases to light and provide additional clues in determining its etiology and pathogenesis. PMID- 11735695 TI - Delayed cutaneous hypersensitivity reactions to hirudin. AB - Hirudin is one of the new synthetic antithrombin agents, which is most commonly used in patients with type II heparin-induced thrombocytopenia and in patients with hypersensitivity reactions to unfractionated heparin as well as low molecular-weight heparins. Hirudin is comparable to heparin as an antithrombotic agent and also has been studied as a primary treatment in patients who experienced acute myocardial infarctions. We describe a patient with a history of type II heparin-induced thrombocytopenia who was placed on intravenous hirudin therapy. After extravasation of the intravenous hirudin site, the patient developed a delayed hypersensitivity reaction that histologically showed an epithelioid granulomatous infiltrate. Although rare reports of hypersensitivity reactions to hirudin have been published, these reactions have not been well characterized and the histopathologic changes have not been described. PMID- 11735696 TI - Unusual clear cell variant of epithelioid mesothelioma. AB - Clear cell mesothelioma is an extremely rare neoplasm of the pleura, which can easily be mistaken for a metastasis of clear cell carcinoma to the pleura. We report here the histochemical, immunohistochemical, and ultrastructural aspects of a new case of clear cell pleural mesothelioma in a 52-year-old man with no known asbestos exposure. He was admitted to the hospital for recurrent pleural effusion, which was negative for neoplastic cells at the cytologic examination. A partial decortication of the right pleura was performed. The morphologic, immunohistochemical, and ultrastructural features reported for this case are consistent with the diagnosis of clear cell mesothelioma. The differential diagnosis and immunohistochemical features in comparison with other clear cell neoplasms are discussed. PMID- 11735697 TI - Serous cystadenoma of the pancreas with papillary features: a diagnostic pitfall on fine-needle aspiration biopsy. AB - Serous cystadenoma of the pancreas is an uncommon neoplasm that occasionally exhibits papillary differentiation. The cytomorphologic structure of pancreatic serous cystadenoma has been rarely described, and, to our knowledge, such papillary morphologic structure has never been reported on fine-needle aspiration cytologic examination. We present a case of serous cystadenoma of the pancreas in a 77-year-old woman. Endoscopic ultrasonography showed a well-demarcated solid/cystic mass in the midbody of the pancreas, suggestive of solid pseudopapillary tumor. Aspiration cytologic examination, performed under endoscopic ultrasound guidance, showed a predominantly papillary epithelial neoplasm consistent with the radiologic impression. Gross and histologic examination of the excised specimen revealed a pancreatic serous cystadenoma with multifocal papillae. This case illustrates the cytomorphologic structure of serous cystadenoma that presents with prominent papillary differentiation on aspiration cytologic examination. The unusual cytologic appearance of this tumor introduces significant diagnostic challenges to the pathologist. Serous cystadenoma must be included in the differential diagnosis of pancreatic neoplasms with papillary morphologic structure as evaluated by fine-needle aspiration cytologic examination. PMID- 11735698 TI - Sialadenoma papilliferum of the hard palate: report of 2 cases and immunohistochemical evaluation. AB - This study reports on the clinical and histologic features of 2 previously unreported cases of sialadenoma papilliferum. Immunohistochemical analysis of one of the cases demonstrated that the ductal cell component shows both epithelial and myoepithelial differentiation. PMID- 11735699 TI - Pulmonary adenocarcinoma simulating malignant mesothelioma. AB - Adenocarcinoma of the lung with pleural involvement frequently resembles pleural epithelioid mesothelioma clinically as well as macro- and microscopically. Special stains, immunohistochemical studies, and electron microscopic studies are needed to differentiate these 2 tumors. We report a case of pleural involvement by adenocarcinoma, mimicking in the hematoxylin-eosin stain an epithelioid mesothelioma, correctly identified only after immunohistochemical and electron microscopic examinations. PMID- 11735700 TI - Primary gastric choriocarcinoma: a case report and review of the literature. AB - Choriocarcinoma is a rapidly invasive, widely metastatic human chorionic gonadotropin (HCG)-producing neoplasm, usually intrauterine and gestational. Primary gastric choriocarcinoma is very rare, and its pathogenesis is still uncertain. We report a case of primary gastric choriocarcinoma associated with adenocarcinoma in a 36-year-old woman. The patient presented with gastrointestinal bleeding and a gastric mass clinically suspicious of gastric adenocarcinoma. Histopathologic evaluation proved the tumor to be a choriocarcinoma, with a minor component of a poorly differentiated adenocarcinoma. The patient was treated with a standard nongestational choriocarcinoma chemotherapy regimen. An impressive initial response was evidenced by clinical reduction of the tumor volume and drop of the serum beta HCG levels after the first cycle. However, the tumor rapidly recurred in the abdomen and disseminated to the lungs, which were documented by new elevation of serum beta-HCG levels and computed tomographic scans despite continuing with 3 more cycles of chemotherapy. The patient died 6 months after diagnosis. PMID- 11735701 TI - Pathologic quiz case: a symptomatic right atrial mass. Papillary fibroelastoma. PMID- 11735702 TI - Pathologic quiz case: a mass of the spleen. Inflammatory myofibroblastic tumor of the spleen. PMID- 11735703 TI - Pathologic quiz case: nonimmune hydrops in a newborn. Down syndrome with acute (transient) leukemia. PMID- 11735704 TI - Pathologic quiz case: lytic lesion in the vertebra of a 17-year-old adolescent girl. PMID- 11735705 TI - Serous microcystic adenoma of the pancreas. PMID- 11735706 TI - Infliximab (anti-tumor necrosis factor alpha antibody): a novel, highly effective treatment of recalcitrant subcorneal pustular dermatosis (Sneddon-Wilkinson disease). PMID- 11735707 TI - Dermoscopic classification of atypical melanocytic nevi (Clark nevi). AB - OBJECTIVES: To create a dermoscopic classification of atypical melanocytic nevi (Clark nevi) and to investigate whether individuals bear a predominant type. DESIGN: Digital dermoscopic images of Clark nevi were classified according to structural features, ie, reticular, globular, or homogeneous patterns or combinations of these types. The nevi were also characterized as central hypopigmented or hyperpigmented, eccentric peripheral hypopigmented or hyperpigmented, or multifocal hypopigmented or hyperpigmented. SETTING: Two pigmented skin lesion clinics. PATIENTS: We examined 829 Clark nevi on 23 individuals. MAIN OUTCOME MEASURE: A reliable dermoscopic classification of Clark nevi and frequency of different dermoscopic types. RESULTS: Using the dermoscopic classification, the 829 Clark nevi were classified as follows: 221 (26.7%) as reticular, 167 (20.1%) as reticular-homogeneous, 148 (17.9%) as globular homogeneous, 112 (13.5%) as reticular-globular, 89 (10.7%) as homogeneous, 84 (10.1%) as globular, and 8 (1.0%) as unclassified. Most individuals were prone to a predominant type of Clark nevus. Seven individuals (30%) showed a single type of Clark nevus in more than 50% of their nevi and 5 (22%) in more than 40% of their nevi. CONCLUSIONS: The proposed dermoscopic classification of Clark nevi is easily applicable and allows a detailed characterization of the different dermoscopic types of Clark nevi. Knowledge of these dermoscopic types should reduce unnecessary surgery for benign melanocytic lesions. Exact classification of the different types of Clark nevi is a necessary prerequisite for further clinical, dermoscopic, and histopathologic studies, which will give new insights in the biology of acquired melanocytic nevi. PMID- 11735708 TI - Short-term digital surface microscopic monitoring of atypical or changing melanocytic lesions. AB - OBJECTIVE: To examine the outcome of short-term digital surface microscopic monitoring of suspicious or changing atypical melanocytic lesions. DESIGN: Digital surface microscopic (oil epiluminescence microscopy, and dermoscopy) images of clinically melanocytic lesions were taken with a color calibrated 3 CCD video instrument. In general, lesions were moderately atypical, flat or only slightly raised, without a history of change or surface microscopic evidence of melanoma, or were mildly atypical lesions with a history of change. Lesions were monitored during a 2.5- to 4.5-month period (median, 3.0 months). With the exception of overall change in pigmentation consistent with that seen in surrounding skin (solar exposure changes), any morphologic change after monitoring was considered an indication to excise. SETTING: Sydney Melanoma Unit, Sydney, Australia (a referral center). PATIENTS: A consecutive sample of 318 lesions from 245 patients (aged 4-81 years). MAIN OUTCOME MEASURE: Specificity for the diagnosis of melanoma. RESULTS: Of the 318 lesions, 81% remained unchanged. Of the 61 lesions that showed morphologic changes, 7 (11% of changed and 2% of total lesions) were found to be early melanoma (5 in situ and 2 invasive with a Breslow thickness of 0.25 mm and 0.28 mm, respectively). None of these melanomas developed any classic surface microscopic features of melanoma and therefore could be identified only by morphologic change. The specificity for the diagnosis of melanoma by means of short-term digital monitoring was 83%. CONCLUSION: On the assumption that all melanoma will change during the monitored period, surface microscopy digital monitoring is a useful adjunct for the management of melanocytic lesions. PMID- 11735709 TI - Risks and benefits of sequential imaging of melanocytic skin lesions in patients with multiple atypical nevi. AB - OBJECTIVE: To evaluate the utility of sequential imaging of melanocytic skin lesions. DESIGN: With the use of a computerized test environment, digital images of 80 melanocytic skin lesions (including 10 early melanomas) were presented to 24 dermatologists with different levels of experience in 3 sessions. The 3 sessions were designed to simulate the decision-making process (1) without the possibility of follow-up, (2) with the possibility of follow-up, and (3) after presentation of follow-up images. MAIN OUTCOME MEASURES: Diagnostic performance in terms of sensitivity, specificity, accuracy, treatment threshold, and utility. RESULTS: The possibility of follow-up increased the treatment threshold in all groups of dermatologists compared with decision making without the possibility of follow-up. The increase of the treatment threshold was accompanied by a loss of sensitivity and a gain in specificity. The overall diagnostic accuracy remained unchanged. After presentation of follow-up images, the diagnostic accuracy improved significantly. The sensitivity improved for all readers, but the specificity improved only for the most experienced readers. The utility of sequential imaging depended on the compliance of patients with follow-up. Under the assumption that all patients are compliant with follow-up, the utility of sequential imaging was superior to decision making without follow-up over a broad range of benefit-risk ratios. CONCLUSIONS: Sequential imaging of melanocytic skin lesions is a useful procedure for patients with multiple atypical nevi. Uncritical use of sequential imaging cannot be recommended, because the utility of this technique depends on the experience in the interpretation of follow-up images and on the patient's compliance with follow-up. PMID- 11735710 TI - Tazarotene cream for the treatment of facial photodamage: a multicenter, investigator-masked, randomized, vehicle-controlled, parallel comparison of 0.01%, 0.025%, 0.05%, and 0.1% tazarotene creams with 0.05% tretinoin emollient cream applied once daily for 24 weeks. AB - OBJECTIVE: To assess the safety and efficacy of 4 concentrations of tazarotene cream in the treatment of facial photodamage. DESIGN: Prospective weekly multicenter, investigator-masked, randomized, parallel-group study. SETTING: University hospitals and clinical research centers. PATIENTS: Three hundred forty nine subjects with facial photodamage. INTERVENTION: Daily topical application of tazarotene cream (0.01%, 0.025%, 0.05%, and 0.1%) compared with its vehicle and with 0.05% tretinoin emollient cream. RESULTS: Tazarotene cream and tretinoin cream significantly improved mottled hyperpigmentation and fine wrinkles. At week 24, treatment success rates based on global responses were 67% (39 of 58 subjects) with 0.1% tazarotene, 52% (30 of 58 subjects) with 0.05% tazarotene, 36% (21 of 58 subjects) with 0.025% tazarotene, 41% (24 of 59 subjects) with 0.01% tazarotene, 55% (32 of 58 subjects) with 0.05% tretinoin, and 22% (13 of 58 subjects) with vehicle. Local adverse events, although more frequent with tazarotene at higher concentrations, were generally mild to moderate. CONCLUSIONS: Tazarotene in a cream formulation is safe and is associated with positive changes in the treatment of photodamaged facial skin. PMID- 11735711 TI - Congenital nonprogressive hemangioma: a distinct clinicopathologic entity unlike infantile hemangioma. AB - BACKGROUND: Infantile hemangiomas are common tumors, distinctive for their perinatal presentation, rapid growth during the first year of life, and subsequent involution-and for their expression of a unique immunophenotype shared by placental microvessels. Occasional "hemangiomas" differ from the classic form in presenting fully formed at birth, then following a static or rapidly involuting course. These congenitally fully developed lesions have generally been assumed to be clinical variants of more typical, postnatally developing hemangiomas. This assumption has not been tested by rigorous histologic and immunophenotypic comparisons. OBJECTIVE: To compare the histologic and immunohistochemical features of congenital nonprogressive hemangiomas with those of typical, postnatally proliferating, hemangiomas. DESIGN: All cellular vascular tumors resected from infants younger than 4 months at Arkansas Children's Hospital, Little Rock, over the past 20 years (43 lesions from 36 patients) were first characterized histologically and immunohistochemically, then clinically by chart review. SETTING: A university-affiliated pediatric hospital. MAIN OUTCOME MEASURES: Histologic appearance, immunoreactivity for the infantile hemangioma associated antigens GLUT1 and LeY, and clinical behavior. RESULTS: Congenital nonprogressive hemangiomas differed from postnatally proliferating infantile hemangiomas in histologic appearance and immunohistochemical profile. Distinguishing pathologic features of these tumors were lobules of capillaries set within densely fibrotic stroma containing hemosiderin deposits; focal lobular thrombosis and sclerosis; frequent association with multiple thin-walled vessels; absence of "intermingling" of the neovasculature with normal tissue elements; and lack of immunoreactivity for GLUT1 and LeY. CONCLUSION: Congenital nonprogressive hemangiomas are histologically and immunophenotypically distinct from classically presenting hemangiomas of infancy, unlikely to be related to the latter in pathogenesis. PMID- 11735712 TI - Idiopathic localized unilateral hyperhidrosis: case report of successful treatment with botulinum toxin type A and review of the literature. AB - BACKGROUND: Localized unilateral hyperhidrosis (LUH) is a rare disorder of unknown origin. We describe a patient with LUH on the forearm, where a fracture was identified as a past injury. OBSERVATIONS: We treated the patient with botulinum toxin type A injections, and he was complaint free during the 6 months after treatment. In addition, the initially strong positive results of the iodine starch test (Minor sweat test) were negative in the affected region after treatment. CONCLUSIONS: This relatively new therapeutic modality already established for axillary, palmar, and plantar hyperhidrosis seems to be efficient in LUH. As the former therapeutic approaches are rather disappointing, and as botulinum toxin type A locally applied shows limited adverse effects, we think a trial of botulinum toxin type A is justified in cases of LUH, even as a first line treatment. In addition, the literature considering localization and causes of LUH is reviewed. PMID- 11735713 TI - A comparison of dermatologists' and primary care physicians' accuracy in diagnosing melanoma: a systematic review. AB - OBJECTIVE: To compare the accuracy of dermatologists and primary care physicians (PCPs) in identifying pigmented lesions suggestive of melanoma and making the appropriate management decision to perform a biopsy or to refer the patient to a specialist. DATA SOURCES: Studies published between January 1966 and October 1999 in the MEDLINE, EMBASE, and CancerLit databases; reference lists of identified studies; abstracts from recent conference proceedings; and direct contact with investigators. Medical subject headings included melanoma, diagnosis, screening, primary care, family practitioner, general practitioner, internal medicine, dermatologist, and skin specialist. Articles were restricted to those involving human subjects. STUDY SELECTION: Studies that presented sufficient data to determine the sensitivity and specificity of dermatologists' or PCPs' ability to correctly diagnose lesions suggestive of melanoma and to perform biopsies on or refer patients with such lesions. DATA EXTRACTION: Two reviewers independently abstracted data regarding the sensitivity and specificity of the dermatologists and PCPs for diagnostic and biopsy or referral accuracy. Disagreements were resolved by discussion. The quality of the studies was also evaluated. DATA SYNTHESIS: Thirty-two studies met inclusion criteria; 10 were prospective studies. For diagnostic accuracy, sensitivity was 0.81 to 1.00 for dermatologists and 0.42 to 1.00 for PCPs. None of the studies reported specificity for dermatologists; one reported specificity for PCPs (0.98). For biopsy or referral accuracy, sensitivity ranged from 0.82 to 1.00 for dermatologists and 0.70 to 0.88 for PCPs; specificity, 0.70 to 0.89 for dermatologists and 0.70 to 0.87 for PCPs. Receiver operating characteristic curves for biopsy or referral ability were inconclusive. CONCLUSIONS: The published data are inadequate to demonstrate differences in dermatologists' and PCPs' diagnostic and biopsy or referral accuracy of lesions suggestive of melanoma. We offer study design suggestions for future studies. PMID- 11735714 TI - A thorough systematic review of treatments for atopic eczema. PMID- 11735715 TI - Exorex for psoriasis: the importance of randomized controlled trials in testing "new" products. PMID- 11735716 TI - The placebo effect: and another one bites the dust. PMID- 11735717 TI - Dermoscopy and early diagnosis of melanoma: the light and the dark. PMID- 11735718 TI - Comparing the diagnostic accuracy of dermatologists and nondermatologists. PMID- 11735719 TI - Papules on the hands. Granuloma annulare. PMID- 11735720 TI - Asymptomatic papules on a child. Perforating granuloma annulare. PMID- 11735721 TI - Symmetrical nasal bridge nodules. Rheumatoid nodule. PMID- 11735722 TI - Erythematous annular plaques in a necklace distribution. Annular elastolytic giant cell granuloma. PMID- 11735723 TI - Speckled lentiginous nevus (nevus spilus): the "roots" of the "melanocytic garden". PMID- 11735724 TI - Speckled lentiginous nevus--classic congenital melanocytic nevus hybrid not the result of "collision". PMID- 11735725 TI - Human papillomavirus type 7 and butcher's warts. PMID- 11735726 TI - The use of ELM to monitor the success of antiscabietic treatment. Epiluminescence light microscopy. PMID- 11735727 TI - Primary cutaneous large B-cell lymphoma of the leg relapsing as cutaneous intravascular large B-cell lymphoma. PMID- 11735728 TI - A prospective open study of topical psoralen-UV-A therapy for Necrobiosis lipoidica. PMID- 11735729 TI - Treatment of refractory, atypical lower extremity ulcers with tissue-engineered skin (Apligraf). PMID- 11735730 TI - A piece of my mind. The consultation. PMID- 11735734 TI - Progress in the treatment of rheumatoid arthritis. PMID- 11735735 TI - At polio's end game, strategies differ. PMID- 11735736 TI - Cardiologists like statins-more than patients do. PMID- 11735741 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735742 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735743 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735744 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735745 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735746 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735747 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735748 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735749 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735751 TI - Cardiovascular events and COX-2 inhibitors. PMID- 11735754 TI - Preventable deaths from medical errors. PMID- 11735752 TI - Risk factors for dating violence in adolescents. PMID- 11735756 TI - Identification and fracture outcomes of undiagnosed low bone mineral density in postmenopausal women: results from the National Osteoporosis Risk Assessment. AB - CONTEXT: Large segments of the population at risk for osteoporosis and fracture have not been evaluated, and the usefulness of peripheral measurements for short term prediction of fracture risk is uncertain. OBJECTIVES: To describe the occurrence of low bone mineral density (BMD) in postmenopausal women, its risk factors, and fracture incidence during short-term follow-up. DESIGN: The National Osteoporosis Risk Assessment, a longitudinal observational study initiated September 1997 to March 1999, with approximately 12 months of subsequent follow up. SETTING AND PARTICIPANTS: A total of 200 160 ambulatory postmenopausal women aged 50 years or older with no previous osteoporosis diagnosis, derived from 4236 primary care practices in 34 states. MAIN OUTCOME MEASURES: Baseline BMD T scores, obtained from peripheral bone densitometry performed at the heel, finger, or forearm; risk factors for low BMD, derived from questionnaire responses; and clinical fracture rates at 12-month follow-up. RESULTS: Using World Health Organization criteria, 39.6% had osteopenia (T score of -1 to -2.49) and 7.2% had osteoporosis (T score /=4 points increase from the baseline National Institutes of Health Stroke Scale [NIHSS] score); clinical outcome (measured by 4 clinical scales) at 3 months, CT lesion volume at 3 months, death at 90 days; and symptomatic intracranial hemorrhage (ICH) within 36 hours of treatment. RESULTS: The prevalence of EIC on baseline CT in the combined rt-PA and placebo groups was 31% (n = 194). The EIC was significantly associated with baseline NIHSS score (rho = 0.23; P<.001) and time from stroke onset to baseline CT scan (rho = 0.11; P =.007). After adjusting for baseline variables, there was no EIC x treatment interaction detected for any clinical outcome, including deterioration at 24 hours, 4 clinical scales, lesion volume, and death at 90 days (P>/=.25), implying that EIC is unlikely to affect response to rt-PA treatment. After adjusting for NIHSS score (an independent predictor of ICH), no EIC association with symptomatic ICH at 36 hours was detected in the group treated with rt-PA (P>/=.22). CONCLUSIONS: Our analysis suggests that EICs are prevalent within 3 hours of stroke onset and correlate with stroke severity. However, EICs are not independently associated with increased risk of adverse outcome after rt-PA treatment. Patients treated with rt-PA did better whether or not they had EICs, suggesting that EICs on CT scan are not critical to the decision to treat otherwise eligible patients with rt-PA within 3 hours of stroke onset. PMID- 11735759 TI - Guided medication dosing for inpatients with renal insufficiency. AB - CONTEXT: Usual drug-prescribing practices may not consider the effects of renal insufficiency on the disposition of certain drugs. Decision aids may help optimize prescribing behavior and reduce medical error. OBJECTIVE: To determine if a system application for adjusting drug dose and frequency in patients with renal insufficiency, when merged with a computerized order entry system, improves drug prescribing and patient outcomes. DESIGN, SETTING, AND PATIENTS: Four consecutive 2-month intervals consisting of control (usual computerized order entry) alternating with intervention (computerized order entry plus decision support system), conducted in September 1997-April 1998 with outcomes assessed among a consecutive sample of 17 828 adults admitted to an urban tertiary care teaching hospital. INTERVENTION: Real-time computerized decision support system for prescribing drugs in patients with renal insufficiency. During intervention periods, the adjusted dose list, default dose amount, and default frequency were displayed to the order-entry user and a notation was provided that adjustments had been made based on renal insufficiency. During control periods, these recommended adjustments were not revealed to the order-entry user, and the unadjusted parameters were displayed. MAIN OUTCOME MEASURES: Rates of appropriate prescription by dose and frequency, length of stay, hospital and pharmacy costs, and changes in renal function, compared among patients with renal insufficiency who were hospitalized during the intervention vs control periods. RESULTS: A total of 7490 patients were found to have some degree of renal insufficiency. In this group, 97 151 orders were written on renally cleared or nephrotoxic medications, of which 14 440 (15%) had at least 1 dosing parameter modified by the computer based on renal function. The fraction of prescriptions deemed appropriate during the intervention vs control periods by dose was 67% vs 54% (P<.001) and by frequency was 59% vs 35% (P<.001). Mean (SD) length of stay was 4.3 (4.5) days vs 4.5 (4.8) days in the intervention vs control periods, respectively (P =.009). There were no significant differences in estimated hospital and pharmacy costs or in the proportion of patients who experienced a decline in renal function during hospitalization. CONCLUSIONS: Guided medication dosing for inpatients with renal insufficiency appears to result in improved dose and frequency choices. This intervention demonstrates a way in which computer based decision support systems can improve care. PMID- 11735760 TI - Epidemic increase in childhood overweight, 1986-1998. AB - CONTEXT: Overweight is the most common health problem facing US children. Data for adults suggest that overweight prevalence has increased by more than 50% in the last 10 years. Data for children also suggest that the prevalence of overweight continues to increase rapidly. OBJECTIVE: To investigate recent changes in the prevalence of overweight within a nationally representative sample of children. DESIGN, SETTING, AND PARTICIPANTS: The National Longitudinal Survey of Youth, a prospective cohort study conducted from 1986 to 1998 among 8270 children aged 4 to 12 years (24 174 growth points were analyzed). MAIN OUTCOME MEASURES: Prevalence of overweight children, defined as body mass index (BMI) greater than the 95th percentile for age and sex, and prevalence of overweight and at-risk children, defined as BMI greater than the 85th percentile for age and sex. The roles of race/ethnicity, sex, income, and region of residence were also examined. RESULTS: Between 1986 and 1998, overweight increased significantly and steadily among African American (P<.001), Hispanic (P<.001), and white (P =.03) children. By 1998, overweight prevalence increased to 21.5% among African Americans, 21.8% among Hispanics, and 12.3% among non-Hispanic whites. In addition, overweight children were heavier in 1998 compared with 1986 (P<.001). After adjusting for confounding variables, overweight increased fastest among minorities and southerners, creating large demographic differences in the prevalence of childhood overweight by 1998. The number of children with BMI greater than the 85th percentile increased significantly from 1986 to 1998 among African American and Hispanic children (P<.001 for both) and nonsignificantly among white children (P =.77). CONCLUSIONS: Childhood overweight continues to increase rapidly in the United States, particularly among African Americans and Hispanics. Culturally competent treatment strategies as well as other policy interventions are required to increase physical activity and encourage healthy eating patterns among children. PMID- 11735761 TI - Sex differences in cardiac catheterization: the role of physician gender. AB - CONTEXT: Many studies indicate that women are less likely than men to undergo cardiac procedures after an acute myocardial infarction (AMI), raising concerns of sexual bias in clinical care. However, no data exist regarding the relationship between patient sex, physician sex, and use of cardiac procedures. OBJECTIVE: To determine whether sex differences in cardiac catheterization after AMI were greater when patients were treated by male attending physicians compared with female attending physicians. DESIGN, SETTING, AND PATIENTS: Analysis of data from the Cooperative Cardiovascular Project, a retrospective medical record review. A total of 104 >231 Medicare fee-for-service beneficiaries who were hospitalized in US acute care hospitals for an AMI between January 1994 and February 1995. MAIN OUTCOME MEASURE: Use of cardiac catheterization within 60 days of admission, compared between the 4 groups of patient sex-physician sex combinations. RESULTS: Women underwent fewer cardiac catheterizations than men when treated by either male physicians (38.6% vs 50.8%; P =.001) or female physicians (34.8% vs 45.8%; P =.001). Sex differences in procedure use were not greater when a patient and physician were of different sexes (P for interaction =.85). After potential confounders in multivariable analysis were accounted for, women were less likely to undergo cardiac catheterization (risk ratio, 0.90 [95% confidence interval (CI), 0.88-0.92]), regardless of the treating physician's sex. Patients treated by male physicians were more likely to undergo cardiac catheterization (risk ratio, 1.06 [95%CI, 1.02-1.10]) than those treated by female physicians, regardless of patient sex. CONCLUSIONS: Women who have had an AMI undergo a cardiac catheterization less often than men, whether treated by a male or female physician. These results suggest that factors other than sexual bias by male physicians toward women account for sex differences in cardiac procedure use. PMID- 11735762 TI - Psychiatric issues in the management of patients with HIV infection. AB - Approximately 1 million persons are now infected with human immunodeficiency virus (HIV) in the United States. Evidence exists that psychiatric disorders are common in patients with HIV and that these patients may not receive optimal care because their psychiatric disorders are a barrier to medical care, communication with clinicians, and adherence to medical recommendations. We describe herein a complex case seen at The Johns Hopkins Hospital with several psychiatric conditions that are common in our HIV clinic population. We describe the collaborative treatment of the patient by a multidisciplinary team including both medical and mental health practitioners. We briefly describe a coherent diagnostic and treatment approach to patients in HIV clinics and the supporting rationale from the literature. We discuss the need for comprehensive evaluation, a multidisciplinary treatment team, and therapeutic optimism. PMID- 11735763 TI - Osteoporosis, an underdiagnosed disease. PMID- 11735764 TI - Improving drug use in elderly patients: getting to the next level. PMID- 11735770 TI - Surgery for Parkinson disease and other movement disorders: benefits and limitations of ablation, stimulation, restoration, and radiation. PMID- 11735771 TI - Investigating chemokines and chemokine receptors in patients with multiple sclerosis: opportunities and challenges. AB - Multiple sclerosis is an autoimmune demyelinating disease of the human central nervous system with an unknown etiology. Crucial to its pathogenesis is the accumulation and activation of mononuclear cells in the central nervous system. Chemokines and their receptors are proposed to play a major role in the inflammatory recruitment of leukocytes. Besides analyses of relationships between chemokine or chemokine receptor gene polymorphisms and multiple sclerosis susceptibility and severity, analyses of chemokines and their receptors in patients with multiple sclerosis remain descriptive. In clinical material, chemokines and chemokine receptors can be examined in body fluids (blood and cerebrospinal fluid) and in brain tissues obtained via biopsy or autopsy. Research results will be summarized in this review, and a general model of leukocyte migration into the central nervous system under normal and inflammatory conditions will be proposed. Furthermore, opportunities and challenges for future investigations will be identified. PMID- 11735772 TI - Current concepts in mild cognitive impairment. AB - The field of aging and dementia is focusing on the characterization of the earliest stages of cognitive impairment. Recent research has identified a transitional state between the cognitive changes of normal aging and Alzheimer's disease (AD), known as mild cognitive impairment (MCI). Mild cognitive impairment refers to the clinical condition between normal aging and AD in which persons experience memory loss to a greater extent than one would expect for age, yet they do not meet currently accepted criteria for clinically probable AD. When these persons are observed longitudinally, they progress to clinically probable AD at a considerably accelerated rate compared with healthy age-matched individuals. Consequently, this condition has been recognized as suitable for possible therapeutic intervention, and several multicenter international treatment trials are under way. Because this is a topic of intense interest, a group of experts on aging and MCI from around the world in the fields of neurology, psychiatry, geriatrics, neuropsychology, neuroimaging, neuropathology, clinical trials, and ethics was convened to summarize the current state of the field of MCI. Participants reviewed the world scientific literature on aging and MCI and summarized the various topics with respect to available evidence on MCI. Diagnostic criteria and clinical outcomes of these subjects are available in the literature. Mild cognitive impairment is believed to be a high-risk condition for the development of clinically probable AD. Heterogeneity in the use of the term was recognized, and subclassifications were suggested. While no treatments are recommended for MCI currently, clinical trials regarding potential therapies are under way. Recommendations concerning ethical issues in the diagnosis and the management of subjects with MCI were made. PMID- 11735773 TI - Complications of gamma knife surgery for Parkinson disease. AB - BACKGROUND: Many medical centers throughout the world offer radiosurgery with the gamma knife (GK) for pallidotomy and thalamotomy as a safe and effective alternative to radiofrequency ablative surgery and deep brain stimulation for Parkinson disease (PD). The reported incidence of significant complications varies considerably, and the long-term complication rate remains unknown. DESIGN: We describe 8 patients seen during an 8-month period referred for complications of GK surgery for PD. RESULTS: Of the 8 patients, 1 died as a result of complications, including dysphagia and aspiration pneumonia. Other complications included hemiplegia, homonymous visual field deficit, hand weakness, dysarthria, hypophonia, aphasia, arm and face numbness, and pseudobulbar laughter. In all patients, lesions were significantly off target. CONCLUSIONS: The 8 patients with PD seen in referral at our center for complications of GK surgery highlight a spectrum of potential problems associated with this procedure. These include lesion accuracy and size and the delayed development of neurological complications secondary to radiation necrosis. Gamma knife surgery may have a higher complication rate than has been previously appreciated due to delayed onset and underreporting. We believe that the risk-benefit ratio of the GK will require further scrutiny when considering pallidotomy or thalamotomy in patients with PD. Physicians using this technique should carefully follow up patients postoperatively for delayed complications, and fully inform patients of these potential risks. PMID- 11735774 TI - Intravenous tissue-type plasminogen activator therapy for ischemic stroke: Houston experience 1996 to 2000. AB - CONTEXT: Intravenous tissue-type plasminogen activator (tPA) therapy using the National Institute of Neurological Disorders and Stroke criteria has been given with variable safety to less than 5% of the patients who have ischemic strokes nationwide. Our center is experienced in treating large numbers of stroke patients with intravenous tPA. OBJECTIVE: To report our total 4-year experience in the treatment of consecutive patients who had an ischemic stroke. DESIGN: Prospective inception cohort registry of all patients seen by our stroke team and an additional retrospective medical record review of all patients treated between January 1, 1996, and June 1, 2000. SETTING: A veteran stroke team composed of fellows and stroke-specialty faculty servicing 1 university and 3 community hospitals in a large urban setting. PATIENTS: Consecutive patients with ischemic stroke treated within the first 3 hours of symptom onset. INTERVENTION: According to the National Institute of Neurological Disorders and Stroke protocol, 0.9 mg/kg of intravenous tissue-type plasminogen activator was administered. MAIN OUTCOME MEASURES: Number and proportion treated, patient demographics, time to treatment, hemorrhage rates, and clinical outcome. RESULTS: A total of 269 patients were treated between January 1, 1996, and June 1, 2000. Their mean age was 68 years (age range, 24-93 years); 48% were women. This represented 9% of all patients admitted with symptoms of cerebral ischemia at our most active hospital (over the final 6 months, 13% of all patients with symptoms of cerebral ischemia and 15% of all acute ischemic stroke patients). Before treatment the mean +/- SD National Institutes of Health Stroke Scale (NIHSS) score was 14.4 +/- 6.1 points (median, 14 points; range, 4-33 points). A tPA bolus was given at 137 minutes (range, 30-180 minutes); 28% of the patients were treated within 2 hours. The mean door-to-needle time was 70 minutes (range, 10-129 minutes). The symptomatic intracerebral hemorrhage rate was 5.6% of those patients with a second set of brain scans (4.5% of all patients), with a declining trend from 1996 to 2000. Protocol violations were found in 13% of all patients; the symptomatic intracerebral hemorrhage rate in these patients was 15%. At 24 hours, the NIHSS score was 10 +/- 8 points (median, 8 points; range, 0-36 points). In-hospital mortality was 15% and the patients' discharge NIHSS scores were 7 +/- 7 points (median, 3 points; range, 0-35 points). CONCLUSIONS: Intravenous tPA therapy can be given to up to 15% of the patients with acute ischemic stroke with a low risk of symptomatic intracerebral hemorrhage. Successful experience with intravenous tPA therapy depends on the experience and organization of the treating team and adherence to published guidelines. PMID- 11735775 TI - Silent infarcts in children with sickle cell anemia and abnormal cerebral artery velocity. AB - BACKGROUND: A substantial minority of neurologically normal children with sickle cell disease have lesions consistent with cerebral infarction as seen on magnetic resonance imaging (MRI). OBJECTIVES: To determine if transfusion therapy affects the rate at which silent infarcts develop and to evaluate the contribution of MRI of the brain to stroke prediction by transcranial Doppler (TCD) ultrasonography. STUDY DESIGN: Children with elevated TCD ultrasonographic velocity were randomized to receive long-term transfusion therapy or standard care. Magnetic resonance imaging of the brain was obtained at randomization, annually, and with clinical neurologic events. The risk for new silent lesions and/or stroke was compared for each treatment arm. RESULTS: Among the 37% of subjects with silent infarcts, those receiving standard care were significantly more likely to develop new silent lesions or stroke than were those who received transfusion therapy. For subjects receiving standard care, those with lesions at baseline were significantly more likely to develop stroke or new silent lesions than those whose MRI studies showed no abnormality. CONCLUSIONS: Transfusion therapy lowers the risk for new silent infarct or stroke for children having both abnormal TCD ultrasonographic velocity and silent infarct. However, those with both abnormalities who are not provided transfusion therapy are at higher risk for developing a new silent infarct or stroke than are those whose initial MRI showed no abnormality. The finding of a silent infarct reinforces the need for TCD ultrasonographic screening and consideration of transfusion therapy if the abnormalities are seen. Similarly, elevated TCD ultrasonographic velocity warrants MRI of the brain because children with both abnormalities seem to be at increased risk for developing new silent infarct or stroke. PMID- 11735776 TI - Correlation between Abetax-40-, Abetax-42-, and Abetax-43-containing amyloid plaques and cognitive decline. AB - CONTEXT: Accumulation of senile plaques containing amyloid beta (Abeta)-protein is a pathologic hallmark of Alzheimer disease. Amyloid beta-peptide is heterogeneous, with carboxyterminal variants ending at residues Val40 (Abetax 40), Ala42 (Abetax-42), or Thr43 (Abetax-43). The relative importance of each of these variants in dementia or cognitive decline remains unclear. OBJECTIVE: To study whether Abeta deposition correlates with dementia and occurs at the earliest signs of cognitive decline. DESIGN, SETTING, AND PATIENTS: Postmortem cross-sectional study comparing the deposition of Abeta variants in the prefrontal cortex of 79 nursing home residents having no, questionable, mild, moderate, or severe dementia. MAIN OUTCOME MEASURES: Levels of staining of Abeta peptides ending at amino acid 40, 42, or 43 in the frontal cortex, as a function of Clinical Dementia Rating score. RESULTS: There were significant deposits of all 3 Abeta species that strongly correlated with cognitive decline. Furthermore, deposition of Abetax-42 and Abetax-43 occurred very early in the disease process before there could be a diagnosis of Alzheimer disease. Levels of deposited Abetax-43 appeared surprisingly high given the low amounts synthesized. CONCLUSIONS: These data indicate that Abetax-42 and Abetax-43 are important species associated with early disease progression and suggest that the physiochemical properties of the Abeta species may be a major determinant in amyloid deposition. The results support an important role for Abeta in mediating initial pathogenic events in Alzheimer disease dementia and reinforce that treatment strategies targeting the formation, accumulation, or cytotoxic effects of Abeta should be pursued. PMID- 11735777 TI - The influence of education on clinically diagnosed dementia incidence and mortality data from the Kungsholmen Project. AB - BACKGROUND: The relationship between education and Alzheimer disease (AD) or dementia has been widely examined and the evidence obtained is mixed. Several hypotheses have been proposed to explain the observed association between them. OBJECTIVE: To further understand the relationship between education and incidence of clinically diagnosed AD or dementia. SUBJECTS AND METHODS: A community-based, dementia-free cohort of 1296 aged 75 years and older was followed up to detect incident AD or dementia cases using Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria. The vital status of all subjects who underwent the clinical examination at follow-up (n = 983) was ascertained for 5 years further. Data were analyzed with Cox proportional hazards model after adjustment for main potential confounders. RESULTS: Over an average (SD) of 2.8 (1.0) years of follow-up, 147 subjects were diagnosed as having dementia (109 subjects as having AD). Among those who were clinically examined at follow-up, 88 died with dementia (68 died with AD) within 5 years. Subjects with a low level of education (<8 vs > or =8 years) had a relative risk of 2.6 (95% confidence interval, 1.5-4.4) for AD and 1.7 (95% confidence interval, 1.1-2.6) for dementia. A low educational level was significantly related to all-cause mortality (relative risk, 1.3; 95% confidence interval, 1.0-1.7; P<.05), but not to the mortality of subjects with AD (relative risk, 1.1; 95% confidence interval, 0.5-2.2) or dementia (relative risk, 0.9; 95% confidence interval, 0.5 1.5). CONCLUSIONS: A low level of education is related to an increased incidence of clinical AD or dementia, but not to the mortality of subjects with AD or dementia. These findings can be accounted for by the "cognitive reserve" hypothesis. Alternatively, the observed association between educational level and incidence of AD or dementia may partly reflect detection bias, by which subjects with a low level of education tend to be clinically diagnosed at an earlier point in time. PMID- 11735778 TI - Oligoclonal band number as a marker for prognosis in multiple sclerosis. AB - The natural course of disease in multiple sclerosis varies. Multiple sclerosis that is clinically apparent but causes minimal disability over time has been labeled benign multiple sclerosis. The ability to predict the subsequent clinical course of multiple sclerosis on the basis of clinical and other supportive data at presentation would be invaluable. In this article we report our findings based on a retrospective analysis of 1800 patients diagnosed as having multiple sclerosis, of which 44 patients met our inclusion criteria. There was a suggestion that a low or absent number of oligoclonal bands in the cerebrospinal fluid at the time of diagnosis predicts a better prognosis. However, quantification of oligoclonal bands in cerebrospinal fluid remains an insensitive prognostic indicator and must not be used to influence decisions regarding therapeutic options. PMID- 11735779 TI - Magnetic resonance spectroscopic imaging in temporal lobe epilepsy: neuronal dysfunction or cell loss? AB - BACKGROUND: Magnetic resonance spectroscopy (MRS) has demonstrated consistent metabolic abnormalities in temporal lobe epilepsy. The reason for decreases in N acetylated compounds are thought to be related to neuronal hippocampal cell loss as observed in hippocampal sclerosis. However, mounting evidence suggest that the N-acetylated compound decreases may be functional and reversible. OBJECTIVE: To establish whether the metabolic changes measured by MRS correlate to hippocampal cell loss in temporal lobe epilepsy. SUBJECTS AND METHODS: We prospectively performed quantitative hippocampal MR imaging volumetry and MRS imaging in 33 patients with intractable mesial temporal lobe epilepsy who were undergoing surgery. A neuronal-glial ratio of cornu ammonis and fascia dentata was obtained and correlated while validating the pathologic analysis by comparisons with specimens of age-matched autopsy control-case hippocampus (n = 14). RESULTS: The neuronal-glial ratio of the patient group was statistically significantly lower than in the control group for the cornu ammonis region (P<.001). Correlations of hippocampal volumes with cornu ammonis and neuronal-glial ratios revealed a significant interdependence (P<.01). However, correlations of the resected hippocampal creatine-N-acetylated compound ratio with the cornu ammonis or fascia dentata neuronal-glial ratios showed no significant interdependence (P>.8). CONCLUSIONS: Our findings support the concept that the metabolic dysfunction measured by MRS imaging and the hippocampal volume loss detected by MR imaging volumetry do not have the same neuropathologic basis. These findings suggest that the MRS imaging metabolic measures reflect neuronal and glial dysfunction rather than neuronal cell loss as previously assumed. PMID- 11735780 TI - Provocative techniques should be used for the diagnosis of psychogenic nonepileptic seizures. PMID- 11735781 TI - Provocative testing should not be used for nonepileptic seizures. PMID- 11735782 TI - The confluence of quality of care, cost-effectiveness, pragmatism, and medical ethics in the diagnosis of nonepileptic seizures: a provocative situation for neurology. PMID- 11735783 TI - Patterns on cranial magnetic resonance imaging in elderly people and vascular disease outcomes. PMID- 11735784 TI - Clear corneal wound infection after phacoemulsification. AB - OBJECTIVE: To evaluate clear corneal wound infections after phacoemulsification. MATERIALS AND METHODS: The medical records of 7 patients with clear corneal wound infections after phacoemulsification were reviewed retrospectively. Data that were reviewed included patient age, sex, onset of symptoms and signs after surgery, possible risk factors for infection, concomitant ocular disease, use of perioperative prophylactic antibiotics and steroids, culture and antibiotic sensitivity results, treatment regimen, and outcome. RESULTS: The median onset of signs and symptoms after surgery was 10 days (range, 4-60 days). Corneal cultures yielded methicillin-resistant Staphylococcus aureus in 2 cases, Streptococcus pneumoniae in 1 case, and Staphylococcus epidermidis in 1 case. Cultures yielded no microorganisms for 1 patient. Corneal cultures were not obtained in 2 patients. In 3 of the 4 culture-positive cases, the isolated microorganisms were resistant to the perioperative prophylactic antibiotics (fluoroquinolones and tobramycin) that were used. No possible risk factors were noted except use of topical steroids 4 times a day without antibiotic coverage for iritis before referral in one of our patients. Six of these 7 wound infections healed with topical antibiotic therapy. One of the patients required lamellar keratectomy and conjunctival flap for complete healing. In 4 of the 7 cases, best-corrected visual acuity at the last follow-up visit was better than 20/40. CONCLUSIONS: Clear corneal wound infection after phacoemulsification is a serious complication of cataract surgery. Infections are caused by gram-positive organisms sensitive to bacitracin and the combination of trimethoprim-sulfamethoxazole but often resistant to aminoglycosides and/or fluoroquinolones. PMID- 11735785 TI - Relationships between laser flare photometry values and complications of uveitis. AB - OBJECTIVE: To determine whether relationships exist between elevated laser flare photometry values and common abnormalities and complications associated with uveitis. METHODS: We retrospectively studied all patients with uveitis on whom laser flare photometry measurements ("flare") were obtained (N = 111) at 2 academic medical centers. The first laser flare photometry values obtained for each patient were compared with the presence or absence of the following abnormalities or complications associated with uveitis: keratic precipitates, posterior synechiae, cataract, macular edema, optic disc edema, and glaucoma. In bilateral cases, the eye with the higher flare was used in primary analyses. RESULTS: Flare was significantly higher in patients with posterior synechiae (P<.001) and in those with macular edema (P =.02) than in patients with uveitis who did not have these complications. Flare was significantly higher in patients with prior cataract surgery or cataract at the study visit than in those without cataracts (P =.001). There was no significant difference in flare between patients with and without keratic precipitates, optic disc edema, or glaucoma. No relationships were found between abnormalities or complications and the level of inflammatory cells or flare as determined by clinical assessment. We also identified an inverse relationship between flare and visual acuity that was not completely explained by the presence of complications in a stepwise regression model. CONCLUSIONS: Although causal relationships were not established, associations between flare and some complications of uveitis suggest that aqueous humor protein may be an important factor in the development of these problems. Consequently, laser flare photometry could play a role in predicting outcomes or monitoring therapy for patients with uveitis. PMID- 11735786 TI - The Advanced Glaucoma Intervention Study: 8. Risk of cataract formation after trabeculectomy. AB - OBJECTIVES: To compare the risk of cataract formation in eyes with and without prior trabeculectomy and to assess other risk factors for cataract. METHODS: The Advanced Glaucoma Intervention Study (AGIS) has been following 789 eyes in 591 patients with medically uncontrolled open-angle glaucoma. From 1988 to 1992, these eyes were randomly assigned to either an argon laser trabeculoplasty (ALT) trabeculectomy-trabeculectomy treatment sequence or a trabeculectomy-ALT trabeculectomy sequence. Cox regression analyses were used to assess risk factors for cataract formation during 7 to 11 years of follow-up. MAIN OUTCOME MEASURES: Cataract, defined as either having had cataract surgery or confirmed severe lens opacity with a best-corrected Early Treatment Diabetic Retinopathy Study visual acuity score less than 65 letters (worse than 20/50). RESULTS: Data are presented on the expected 5-year cumulative probability of cataract formation in each randomized sequence by age and presence of diabetes at study entry. Overall, approximately half of the eyes studied developed cataract. A first trabeculectomy, whether as the first or second AGIS intervention, increased the overall risk of cataract by 78% (risk ratio [RR] = 1.78; P<.001). Diabetes (RR = 1.47; P =.004) and age at study entry (RR = 1.07 per year of age; P<.001) were also risk factors for cataract. When postoperative complications of trabeculectomy were included in the analysis, the increased risk of cataract for eyes with a first trabeculectomy reduced to 47% when complications did not occur (RR = 1.47; P =.003) and increased to 104% when complications did occur (RR = 2.04; P<.001). Several specific postoperative complications of trabeculectomy were associated with increased risk of cataract, particularly marked inflammation (RR = 3.29; P<.001) and flat anterior chamber (RR = 1.80; P =.004). Trabeculectomy with complications was also significantly associated with an increased risk of cataract in each of 3 lens regions: nuclear, cortical, and posterior subcapsular. CONCLUSIONS: In eyes of AGIS patients, after adjustment for age and diabetes, trabeculectomy increased the risk of cataract formation by 78%. PMID- 11735787 TI - Analysis of retinal and choroidal circulation during central retinal vein occlusion using indocyanine green videoangiography. AB - OBJECTIVE: To explore the hemodynamic changes and their correlation with clinical presentation during central retinal vein occlusion. MATERIALS AND METHODS: Retrospective, 2-center study. The medical records of 27 patients with central retinal vein occlusion were reviewed. For each patient, the plasma transit in central retinal vessels and in peripapillary choroidal vessels was analyzed using indocyanine green videoangiography. RESULTS: The incidence of alteration in retinal plasma transit, ie, pulsatile filling of arteries and/or veins and increased arteriovenous filling time, was inversely correlated to duration from the onset but not to funduscopic features. Among the 14 patients with less than 1 month's duration of symptoms, 3 developed chronic macular edema, and impairment of arterial flow preceded its onset. Among the 10 patients with opticociliary circulation, choroidal drainage routes were identified in 5 cases, with pulsatile filling in 3. CONCLUSIONS: Early in the course of central retinal vein occlusion, arterial and/or venous flow alterations are present, irrespective of visual acuity, vein dilation, or fundus hemorrhages. These alterations are less frequent in chronic than in recent-onset central retinal vein occlusion. The mechanisms of these alterations remain uncertain but may involve arterial constriction and/or intermittent venous compression. The relationship between these alterations in retinal flow and the secondary onset of macular edema or capillary nonperfusion deserves further investigation. PMID- 11735788 TI - Prevalence and causes of visual field loss in the elderly and associations with impairment in daily functioning: the Rotterdam Study. AB - OBJECTIVES: To determine the prevalence and causes of visual field loss (VFL) and the association between VFL and indicators of impairment in daily functioning. DESIGN: Population-based cohort study. SETTING: Suburb of Rotterdam, the Netherlands. PARTICIPANTS: Community-dwelling elderly residents (n = 6250). MAIN OUTCOME MEASURE: Visual field loss on suprathreshold static, Goldmann kinetic perimetry, or both. METHODS: Suprathreshold testing of the central visual field was performed on both eyes and repeated if results were abnormal or unreliable. Goldmann perimetry was performed to confirm defects. Causes were determined using ophthalmologic and neurologic examination data and medical records. Impairment was assessed using data from interviews and medical records on disability in daily life, falling, and fractures. RESULTS: The overall prevalence of VFL was 5.6% (3.0% in those aged 55-64 years to 17.0% in those > or =85 years); glaucoma was the leading cause in all age groups. Before age 75 years, other optic nerve diseases and stroke ranked second and third, respectively, as did age-related macular degeneration and retinal vascular occlusive disease, respectively, after this age. Also, after adjustment for visual acuity, VFL was associated with disability, diminished enjoyment of reading and watching television, and a higher risk of incident falling. Risk of incident hip fracture was not increased. CONCLUSIONS: Visual field loss is present in 1 of every 20 community-dwelling elderly people and is associated with impaired daily functioning. Glaucoma is the leading cause in all age groups. Other high-ranking causes, some of which are partly preventable, vary by age. PMID- 11735789 TI - Recently acquired diplopia in adults with long-standing strabismus. AB - BACKGROUND: The evaluation and management of recent-onset diplopia in an adult with a history of long-standing strabismus can be perplexing and challenging. No guidelines exist, to my knowledge, for the examination of such patients. DESIGN: A retrospective medical record review. SUBJECTS: Patients seen in my practice with a history of recently acquired diplopia and a history of strabismus dating back to childhood. RESULTS: One hundred fifty-two patients who met the enrollment criteria were identified. Using the treatment approach outlined herein, 132 patients were relieved of their symptoms of diplopia. In most cases, the onset of the diplopia could be correlated with a change in the patient's ocular alignment, refractive needs, or refractive management. Returning patients to their motor status before the onset of symptoms or addressing the change in refractive needs or management usually resulted in relief of symptoms. CONCLUSION: In most cases, adult patients with a history of long-standing strabismus and a recent onset of diplopia can be effectively treated. PMID- 11735790 TI - Surgical treatment of canalicular stenosis in patients receiving docetaxel weekly. AB - OBJECTIVE: To describe the surgical treatment and outcomes in patients with metastatic breast cancer and canalicular stenosis caused by weekly treatment with docetaxel. METHODS: This case series included 10 patients with persistent epiphora due to weekly docetaxel treatment, who were evaluated by probing and irrigation of the tear drainage apparatus and were found to have significant canalicular stenosis. The severity of canalicular stenosis was graded clinically. Each patient underwent bicanalicular silicone intubation or dacryocystorhinostomy (DCR) with placement of either a silicone tube (canaliculo DCR) or a pyrex glass tube (conjunctivo DCR). RESULTS: Seven patients (12 eyes) underwent bicanalicular silicone intubation. Three patients (5 eyes) required canaliculo DCR with the placement of a silicone tube. In 2 patients (3 eyes), the degree of canalicular stenosis was severe enough to require conjunctivo DCR with the placement of a pyrex glass tube. All 10 patients had complete resolution of epiphora immediately after surgery. Four patients continued to receive docetaxel after surgery. In patients who underwent bicanalicular silicone intubation, the silicone stent was kept in place for the duration of docetaxel therapy. There were no surgical or anesthesia-related complications. At a mean follow-up time of 9 months after surgery, all but 1 patient (1 eye) remained asymptomatic. The interval between initiation of docetaxel therapy and surgery was significantly higher in patients who required DCR compared with those who had silicone intubation. The mean cumulative dose of docetaxel at the time of surgery was higher in patients who required DCR than in patients who had silicone intubation, but this difference was not statistically significant. CONCLUSIONS: Canalicular stenosis secondary to weekly treatment with docetaxel should be treated with bicanalicular silicone intubation early in the course of docetaxel therapy. Failure to treat this adverse effect early may likely lead to severe and irreversible canalicular stenosis, which may necessitate conjunctivo DCR with the placement of a permanent pyrex glass tube. PMID- 11735791 TI - Angiostatin produced by certain primary uveal melanoma cell lines impedes the development of liver metastases. AB - OBJECTIVES: To evaluate the ability of human uveal melanomas to produce angiostatin in vitro and the effect of angiostatin on the development of liver metastases in vivo. METHODS: Human uveal melanoma cell lines (OCM1, OCM3, MEL202, MEL285, 92-1, OM431, and OMM1) were assayed for their ability to produce angiostatin in vitro by an angiostatin bioassay and by Western blot analysis. The OCM3 and OMM1 tumor cells were inoculated either in the posterior or the anterior segment of nude mice. One group of mice in each experiment underwent enucleation and hepatic metastases were assayed by histopathologic and liver function analysis. RESULTS: OCM1, OCM3, and 92-1 cell lines significantly inhibited bovine endothelial cell proliferation in vitro and generated 38-Kd angiostatin molecules. Enucleation of eyes containing OCM3 in the posterior segment resulted in a higher number of metastatic foci (26.5) in that group compared with the nonenucleated group of mice (11.17). After enucleation, elevated levels of serum aspartate transaminase and alanine aminotransferase were observed in mice bearing OCM3 in either anterior or posterior segments. The enucleation of eyes containing OMM1 (nonangiostatin-producing cells) had no significant effect on liver metastasis. CONCLUSION: By removing a source of angiostatin, enucleation of melanoma-containing eyes may unwittingly exacerbate the metastatic potential of uveal melanomas. CLINICAL RELEVANCE: In certain circumstances, enucleating a melanoma-containing eye may unwittingly exacerbate metastatic disease. The results also suggest that exogenous angiostatin may have potential therapeutic implications in the management of patients with primary intraocular melanomas. PMID- 11735792 TI - Depth of penetration of scanning laser Doppler flowmetry in the primate optic nerve. AB - OBJECTIVES: To estimate the measuring depth of the blood flow and to establish the vascular contributions to these measurements with scanning laser Doppler flowmetry (SLDF) of the primate anterior optic nerve. METHODS: Optic nerve blood flow in each eye of 8 monkeys was measured using SLDF before and following surgical occlusion of the central retinal artery (n = 4) or posterior ciliary arteries (n = 4). The regional blood flow in both eyes was determined using a nonradioactive microsphere method. RESULTS: The blood flow in the nerve fiber layer (NFL), including the prelaminar region, was measured with microspheres after central retinal artery occlusion; it was significantly reduced (-83%) with no significant change in the combined laminar and retrolaminar regions. The blood flow measured with SLDF had a 51% reduction. After posterior ciliary artery occlusion, the blood flow in the NFL was measured with microspheres and was not significantly affected (+2%); neither was that measured with SLDF (-12%). However, there was a 51% reduction in the laminar and retrolaminar regions when microspheres were used. The mean +/- SD tissue thickness of the NFL was 359 +/- 16 microm and 353 +/- 54 microm in each group. CONCLUSIONS: Scanning laser Doppler flowmetry measures blood flow principally in the NFL of the anterior optic nerve, which is primarily supplied by the central retinal artery. Blood flow in the laminar and retrolaminar regions makes a small contribution to the SLDF measurement, with an NFL thickness between 300 and 400 microm. CLINICAL RELEVANCE: Scanning laser Doppler flowmetry is used for the noninvasive evaluation of ocular microcirculation in diseases such as glaucoma. Because of the dual blood flow supply in the optic nerve and the limited penetration power of the laser, the instrument primarily measures the microcirculation on the surface of the optic nerve, which is largely supplied by the central retinal artery rather than the ciliary arteries. PMID- 11735793 TI - Augmented reality fundus biomicroscopy: a working clinical prototype. AB - BACKGROUND: To guide treatment for macular diseases and to facilitate real-time image correlation, measurement, and comparison, we developed a method for direct overlay of previously stored photographic and angiographic images onto the real time slitlamp fundus view. METHODS: Previously acquired fundus photographs and angiography images were digitized. A slitlamp interfaced to a charge-coupled device camera, framegrabber, and computer allowed for real-time acquisition and digitization of slitlamp fundus images that was synchronous with posterior segment examination. Custom-developed video injectors containing a miniature cathode ray tube display allowed for real-time superposition of angiographic images to the fundus view. Registration and tracking algorithms were developed and deployed in C++. The feasibility of this approach was demonstrated in 5 human subjects. RESULTS: The computer-vision algorithms provided robust registration, tracking, and image overlay of previously stored photographic and angiographic images directly onto the real-time fundus view. Accurate tracking was demonstrated with updates at 3 to 5 Hz. Direct overlay of previously stored images confirmed registration accuracy, but examiners preferred a more simple rendering that included only relevant information and eliminated extraneous, potentially confusing image data. CONCLUSIONS: Slitlamp-based video injection of previously stored images allows for accurate, robust, real-time correlation and comparison to the biomicroscopic fundus view in human subjects. PMID- 11735794 TI - The prevalence of glaucoma in a population-based study of Hispanic subjects: Proyecto VER. AB - OBJECTIVE: To determine the prevalence of glaucoma in a population-based sample of Hispanic adults older than 40 years. METHODS: Using 1990 census data for Arizona, groups of persons living in sections of the city in Nogales and Tucson were randomly selected with a probability proportional to the Hispanic population older than 40 years. We tried to recruit all eligible adults in homes with 1 self described Hispanic adult. Detailed ocular examinations at a local clinic included visual acuity testing, applanation tonometry, gonioscopy, an optic disc evaluation, and a threshold visual field test. Open-angle glaucoma (OAG) was defined using a proposed international system for prevalence surveys, including threshold visual field defect and optic disc damage. Angle-closure glaucoma was defined as bilateral appositional angle closure, combined with optic nerve damage (judged by field and disc as for OAG). RESULTS: Examinations were conducted in 72% (4774/6658) of eligible persons, with a 1.97% prevalence (95% confidence interval, 1.58%-2.36%) of OAG (94 persons). The age-specific OAG prevalence increased nonlinearly from 0.50% in those aged 41 to 49 years to 12.63% in those 80 years and older. Angle-closure glaucoma was detected in 5 persons (0.10%). Sex, blood pressure, and cigarette smoking were not significant OAG risk factors. Only 36 (38%) of the 94 persons with OAG were aware of their OAG before the study. Screening results with an intraocular pressure higher than 22 mm Hg (in the eye with a higher pressure) would miss 80% of the OAG cases. CONCLUSIONS: The prevalence of OAG in Hispanic persons was intermediate between reported values for white and black persons. The prevalence increased more quickly with increasing age than in other ethnic groups. Glaucoma was the leading cause of bilateral blindness. PMID- 11735795 TI - Frequency and risk factors for pterygium in the Barbados Eye Study. AB - OBJECTIVE: To describe the distribution and risk factors for pterygium in the predominantly black population of the Barbados Eye Study, which was based on a random sample of Barbadian-born citizens between the ages of 40 and 84 years. METHODS: The standardized protocol included ophthalmic and other measurements, automated perimetry, lens gradings, fundus photography, and a detailed interview. A 10% systematic sample of participants and those meeting specific criteria also received a comprehensive ophthalmologic evaluation. RESULTS: The Barbados Eye Study included 4709 participants, of whom 2978 were referred for an ophthalmologic evaluation and 2781 (93%) completed the examination. Cases of pterygium were found among 23.4% of 2617 black, 23.7% of 97 mixed (black and white), and 10.2% of 59 white participants examined. In addition to African ancestry, logistic regression analyses indicated a positive association between pterygium and age (odds ratio [OR], 1.01; 95% confidence interval [CI], 1.00 1.02), fewer years of education (OR, 1.43; 95% CI, 1.01-2.03), and an outdoor job location (OR, 1.87; 95% CI, 1.52-2.29). Having a darker skin complexion (OR, 0.66; 95% CI, 0.52-0.83), always using sunglasses outdoors (OR, 0.18; 95% CI, 0.06-0.59), and the use of prescription glasses (OR, 0.75; 95% CI, 0.60-0.93) were protective factors. CONCLUSIONS: Approximately one quarter of the black participants examined had pterygia, a frequency that was 2.5 to 3 times higher than among whites in the Barbados Eye Study and elsewhere. Pterygium was almost twice as frequent among persons who worked outdoors but was only one fifth as likely among those who always used sunglasses outdoors. Educational interventions to modify these potential exposures may assist in preventing pterygium. PMID- 11735796 TI - Relationship of dietary fat to age-related maculopathy in the Third National Health and Nutrition Examination Survey. AB - OBJECTIVE: To evaluate the associations between dietary fat and age-related maculopathy (ARM) in persons 40 years or older who participated in the Third National Health and Nutrition Examination Survey. METHODS: We used a single, nonmydriatic, fundus photograph of 1 eye to ascertain ARM status in 7883 of 11 448 survey participants. Intake of fat was estimated from 24-hour recall, and specific sources of dietary fat were estimated from responses to food frequency questionnaires. Logistic regression was used to compute odds ratios (ORs) and 95% confidence intervals (CIs) that accounted for complex survey design, nonresponse, and potential risk factors for ARM (age, smoking, race, sex, body mass index, history of cardiovascular disease or hypertension, eye color, and sedentary lifestyle). Persons aged 40 to 79 years (n = 7405) were included in analyses for early ARM (n = 644); those 60 years or older (n = 4294) were included in analyses for late ARM (n = 53). RESULTS: After adjustment for age, race, eye color, and sedentary lifestyle, OR for early ARM was 1.4 (95% CI, 0.9-2.2; P for trend,.10) among persons in high vs low quintiles of total fat intake (percentage of total energy). Associations for specific types of fatty acids (as percentages of caloric intake) were in the same direction and unrelated to ARM. The OR for late ARM was 0.7 (95% CI, 0.2-2.6; P for trend,.60) in persons 60 years or older. Further adjustments for other potential confounders did not significantly affect the ORs. CONCLUSION: Age-related maculopathy was not significantly associated with dietary fat in this large cross-sectional survey. PMID- 11735797 TI - Trichiasis and disability in a trachoma-endemic area of Tanzania. AB - OBJECTIVE: To measure limitations in the daily activities of village life associated with having trichiasis for individuals with and without visual acuity loss. METHODS: Men and women 40 years and older in 6 randomly chosen rural villages in the Kongwa district of Tanzania had visual acuity measured and were examined by an ophthalmologist. Subjects indicated the degree of difficulty with daily activities of village life and whether the difficulty was related, in any way, to vision. Limitations were scored using an indicator of "any difficulty" and using a 4-point scale ranging from "no difficulty" to "unable to do." Scores of individuals with and without trichiasis were compared separately for men and women. RESULTS: Among men, trichiasis was associated with excess functional limitation only for those with visual acuity loss (adjusted difference in proportion of tasks [AD] compared with men with neither trichiasis nor visual impairment, 0.35; 95% confidence interval [CI], 0.23-0.47). For women, trichiasis alone was limiting (AD, 0.15; 95% CI, 0.08-0.22) similarly to visual acuity loss alone (AD, 0.09; 95% CI 0.06-0.13), and the combination led to greater limitations (AD, 0.32; 95% CI, 0.26-0.39). CONCLUSION: The burden of trichiasis is likely greater than previously estimated, especially in women for whom trichiasis alone was disabling. PMID- 11735798 TI - Suzanne Schiff-Wertheimer, MD, as a role model. PMID- 11735799 TI - Human studies, institutional review boards, and the protection of human subjects. PMID- 11735800 TI - Hospitals without eye surgery: are minimum standards of care being met for patients with ocular injury? PMID- 11735801 TI - Lone-star tick bite of the conjunctiva. PMID- 11735802 TI - A case of atypical WAGR syndrome with anterior segment anomaly and microphthalmos. PMID- 11735803 TI - Multiple evanescent white dot syndrome following hepatitis a vaccination. PMID- 11735804 TI - Clinicopathologic reports, case reports, and small case series: 0.01% becaplermin gel for the treatment of a chronic orbital ulcer after exenteration. PMID- 11735805 TI - Clinicopathologic reports, case reports, and small case series: progressive growth of benign adenoma of the pigment epithelium of the ciliary body. PMID- 11735806 TI - Extramedullary myeloid cell tumor in an elderly man. PMID- 11735807 TI - Photo essay: bilateral multiple eyelid apocrine hidrocystomas and ectodermal dysplasia. PMID- 11735808 TI - Photo essay: combined occlusion of the central retinal artery and vein in a pediatric patient secondary to infective endocarditis. PMID- 11735809 TI - Quo vadis otologica? PMID- 11735810 TI - Vibrant semi-implantable hearing device with digital sound processing: effective gain and speech perception. AB - BACKGROUND: The Vibrant Soundbridge (Symphonix Devices, San Jose, Calif) is a semi-implantable hearing device. The transducer is attached directly to the incus and is linked by telemetry to the externally worn audioprocessor. A major advantage of this semi-implantable setup, especially during its experimental phase, is that the audioprocessor can be updated. Recently, we replaced the previous 2-channel analog audioprocessor in 14 patients with a 3-channel digital device. DESIGN: Prospective clinical study. Basic functions were measured, including gain as a function of input level and speech perception in quiet. PATIENTS: Patients (n = 14) had moderate to severe sensorineural hearing impairment (average hearing threshold at 0.5, 1.0, 2.0, and 4.0 kHz of 40- to 76 dB hearing level [HL]) and chronic external otitis, which contraindicated use of an ear mold. RESULTS: Gain of the 3-channel audioprocessor for comfortable listening levels and for conversational levels varied from approximately 15- to 30-dB HL, suggesting that the device is suitable for patients with hearing loss of up to 60- to 70-dB HL. In 5 patients, identical measurements were performed using their conventional hearing aids. The other 9 patients did not use a conventional hearing device because of severe external otitis. On average, results obtained with the Vibrant Soundbridge were not as good as those obtained with the conventional device. Nevertheless, patients were satisfied with the Vibrant Soundbridge because they could use it all day without pain or itching. CONCLUSIONS: The Vibrant Soundbridge is suitable for patients with hearing loss of up to 70-dB HL. Compared with conventional devices, in audiometric terms, a surplus value of the Vibrant Soundbridge was not found. PMID- 11735811 TI - Loss of PTEN expression as a prognostic marker for tongue cancer. AB - BACKGROUND: Abnormalities of PTEN, a candidate tumor suppressor gene located at 10q23.3, play an important role in the tumorigenesis of multiple tumor types. OBJECTIVES: To investigate the expression of PTEN and its clinical implication in squamous cell carcinoma of the tongue. DESIGN: Retrospective analysis of PTEN protein expression in archived primary oral tongue tumor samples. SETTING: Academic center. PATIENTS AND METHODS: PTEN expression was determined by immunohistochemical analysis in tissue samples from 41 patients with stage II, III, and IV squamous cell carcinoma of the tongue. All the patients underwent curative surgical treatment with a median follow-up of 81 months. The Kaplan Meier method was used for survival analysis. Multivariate analysis was performed according to the Cox proportional hazards model. RESULTS: Lack of staining for PTEN was demonstrated in 12 (29%) of the 41 tumors. Patients whose tumors lacked PTEN expression had a significantly shorter overall survival time (P = .03) and event-free survival time (P = .01) than those patients with positive PTEN expression. Multivariate regression analysis demonstrated that PTEN expression is an independent predictor of poor outcome when compared with tumor stage and nodal status. CONCLUSIONS: Although genetic alterations of the PTEN gene are rare in head and neck squamous cell carcinoma, loss of PTEN is not an uncommon event in squamous cell carcinoma of the tongue. Lack of PTEN expression may be an independent prognostic indicator for clinical outcome in patients with this tumor type. PMID- 11735812 TI - Chemoradiation for locally advanced squamous cell carcinoma of the head and neck for organ preservation and palliation. AB - OBJECTIVES: To measure the efficacy and toxic effects of our chemoradiotherapy regimen by means of response and survival in patients with advanced squamous cell carcinoma of the head and neck (HNSCC) for organ preservation in resectable disease or palliation in unresectable disease. DESIGN: All patients underwent evaluation by the multidisciplinary head and neck cancer team, with pathological diagnosis and staging. All patients underwent assessment for response to therapy using results of physical examination and radiologic imaging. Patients were followed up at 3-month intervals for a planned period of 5 years. SETTING: Academic center. PATIENTS: Thirty-eight previously untreated patients with newly diagnosed HNSCC were treated from June 1, 1996, through December 31, 1998, of whom 20 had resectable and 18 had unresectable tumors. INTERVENTION: Patients received intravenous cisplatin, 100 mg/m(2) for 1 hour on days 1 and 29; a 24 hour continuous infusion of fluorouracil, 1000 mg/m(2) on days 1 through 4 and 29 through 32; and radiation therapy, 150 rad twice daily for 12 days. The patients were given a 7- to 10-day break, and radiation therapy was restarted on day 29 for 12 additional days (total dose, 7200 rad). MAIN OUTCOME MEASURES: Complete, partial, and total response rates; disease-free survival; overall survival; and toxic effects. RESULTS: Toxic effects of treatment were moderately severe, including grades III to IV mucositis (89%), neutropenia (71%), and renal toxic effects (8%). In the 18 patients in the unresectable group, complete response in the 17 primary tumors and 15 cervical nodal metastases was achieved in 12 (71%) and 9 (60%), respectively; in the 20 patients undergoing organ preservation, complete response rates were 100% in the 23 primary tumors and 15 cervical nodal metastases. Complete response for all 38 patients was achieved in 31 (82%). In the unresectable group, the Kaplan-Meier relapse-free survival estimate is 56%, with follow-up from 29 to 45 months. In the organ preservation group, 75% of patients are alive without disease, and 8 have been followed up for 36 to 48 months. Of the 5 patients who have died, only 2 died of disease, with recurrences at 13.0 and 16.5 months. CONCLUSIONS: Chemoradiotherapy consisting of cisplatin, fluorouracil, and twice-daily external beam radiation is highly effective in achieving durable complete responses in patients with resectable HNSCC undergoing organ preservation and patients with unresectable HNSCC undergoing palliation. Toxic effects of this regimen were moderate to severe. PMID- 11735813 TI - Bone or cartilage invasion by advanced head and neck cancer: intra-arterial supradose cisplatin chemotherapy and concomitant radiotherapy for organ preservation. AB - BACKGROUND: Invasion of bony or cartilaginous structures by advanced upper aerodigestive tract cancer has been considered an indication for surgery on the basis of historic experience of poor responsiveness to radiation therapy. At University of Tennessee-Memphis, patients with advanced head and neck cancer have been treated on a protocol of concomitant intra-arterial (targeted) cisplatin and conventional radiation therapy. OBJECTIVE: To compare the efficacy, in terms of disease control and survival, of this protocol in patients with T4 squamous cell cancers and invasion of bony or cartilaginous structures (group 1; n = 45) vs those with T4 disease but no bone or cartilage involvement (group 2; n = 90). DESIGN: Subset analysis of protocol database and retrospective chart review. METHODS: Treatment consisted of 4 weekly intra-arterial infusions of cisplatin (150 mg/m(2) per week), with simultaneous systemic neutralization by intravenous sodium thiosulfate (9 mg/m(2)), and concurrent radiation therapy at 180 rad (1.8 Gy) or 200 rad (2 Gy) per fraction to a planned total of 6600 to 7400 rad (66-74 Gy) to the primary site or overt nodal disease. Presence of bone or cartilage invasion was established by review of tumor diagrams of clinical findings and computed tomography or magnetic resonance imaging reports. RESULTS: Of 135 patients who had T4 disease and a minimum follow-up of 9 months (median, 40 months), 45 had clinical or radiologic evidence of bone (n = 29: mandible, 12; maxilla, 9; sphenoid, 3; hyoid, 6) and/or cartilage (n = 18: thyroid, 16; cricoid, 4) invasion (some patients had involvement of more than 1 site). The rate of complete response in group 1 (66.7%) was not significantly different from that in group 2 (71.1%) (chi(2) test, P = .79). The 2-year overall actuarial survival for group 1 (46.3%; 95% confidence interval, 30.3%-62.3%) was not significantly different (generalized Wilcoxon test, P = .36) from that of group 2 (36.9%; 95% confidence interval, 25.5%-48.4%). A marked trend was noted for higher response rates in cases of cartilage invasion (81.2%) than in those with bone invasion (58.6%) (P = .15). CONCLUSION: Equivalent efficacy of treatment in the 2 groups suggests that targeted chemoradiation can be a definitive therapeutic option in patients with advanced head and neck cancer invading bony or cartilaginous structures. PMID- 11735814 TI - Management of extensive cervical nodal metastasis in nasopharyngeal carcinoma after radiotherapy: a clinicopathological study. AB - OBJECTIVES: To evaluate the efficacy of afterloading brachytherapy following radical neck dissection (RND) in the management of extensive cervical lymph node disease in nasopharyngeal carcinoma after radiotherapy; and to examine prospectively prognostic factors and the pathologic behavior of neck disease. PATIENTS: Twenty-seven patients with nasopharyngeal carcinoma who had extensive cervical lymph node metastasis following external radiotherapy were treated with RND. Thirteen of them also underwent afterloading brachytherapy with iridium wire (Ir 192). The RND specimens of the 27 patients were also examined with step serial whole-specimen sectioning. RESULTS: All patients survived and their wounds healed primarily. Pathologic examination revealed 183 tumor-bearing lymph nodes that contained tumors in the neck: level I, 4% (8/183); level II, 53% (96/183); level III, 34% (62/183); level IV, 5% (9/183); and level V, 4% (8/183). Extracapsular tumor extension was seen in 84% of patients. Multivariate analysis identified the number of tumor-bearing lymph nodes detected in the specimens to be the only significant factor that affected control of disease. Although the neck disease in the group of patients who had afterloading brachytherapy was more extensive, the 3-year actuarial tumor control for the groups with and without brachytherapy were 60% and 61%, respectively. CONCLUSIONS: Recurrent cervical lymph nodes after radiotherapy in nasopharyngeal carcinoma are extensive and RND is mandatory for a successful salvage. When the nodal metastasis infiltrate or adhere to surrounding tissue, afterloading brachytherapy with iridium wire can provide satisfactory local tumor control. PMID- 11735815 TI - Sensate radial forearm free flaps in tongue reconstruction. AB - BACKGROUND: Successful rehabilitation after ablative surgery requires not only the reconstruction of 3-dimensional form but also the restoration of physiologic function. OBJECTIVE: To assess sensory recovery of reinnervated radial forearm flaps used for tongue reconstruction. PATIENTS AND METHODS: Seventeen patients, who underwent reconstruction of glossectomy defects with reinnervated radial forearm free flaps, formed the study group. Recovery of sensation was measured by both subjective and detailed objective tests 8 months after surgery. Sensory function of the flap was compared with that of the normal residual tongue or the adjacent oral mucosa and the contralateral forearm donor site. RESULTS: All patients involved in this study had tongue defects of hemiglossectomy or greater and adjacent floor of the mouth. Sensory recovery was observed in all of the 17 patients within 8 months. Detailed sensory testing showed that median static 2 point discrimination, moving 2-point discrimination, and pressure sensitivity (1.2 cm, 0.8 cm, and 3.7 psi, respectively) were subjectively greater in the innervated forearm flaps than in the contralateral forearm donor site (2.3 cm, 1.7 cm, and 4.6 psi, respectively) (P= .064) and similar to those of the normal tongue (0.9 cm, 0.5 cm, and 3.6 psi). CONCLUSIONS: In all modalities examined, sensate free flaps proved superior in sensory fidelity to the native forearm donor site and closely approached that of the normal tongue. Microsurgical reinnervation of flaps should be considered in tongue reconstruction. PMID- 11735816 TI - The mucosal invasion model: a novel in vitro model for evaluating the invasive behavior of mucocutaneous malignancies. AB - BACKGROUND: Prevention of regional and metastatic spread of cutaneous malignancies requires understanding the physiologic mechanism of tumor cell invasion. In vitro models are convenient for studying the in vitro invasive phenotype of normal cells, tumor cell lines, or genetically altered cells in a 3 dimensional matrix, but they should attempt to recapitulate the complex in vivo submucosal environment. A new acellular extracellular matrix, porcine submucosal matrix (PSM), is thought to accurately recapitulate the submucosal matrix. A novel in vitro model using PSM to assess mucocutaneous tumor cell invasion was studied. METHODS: The morphologic characteristics, growth, and invasive behavior of human head and neck squamous cell carcinoma (UM-SCC-1, UM-SCC-5, UM-SCC-17B, and OSC-19) cell lines were assessed on the PSM gel and compared with commonly used in vitro invasion models (type I collagen and Matrigel matrices). The invasive phenotype of canine kidney cells was also assessed on each matrix, because this cell line is known to demonstrate a characteristic in vitro invasive phenotype. RESULTS: The PSM-supported head and neck squamous cell carcinoma tumor cell line growth and single cell invasion were seen under stimulated conditions, similar to type I collagen gels. The invasive phenotype of canine kidney cells behaved similarly on PSM and collagen. Matrigel did not support growth well, and invasion occurred only superficially in isolated areas. CONCLUSION: The PSM is a good in vitro model for assessment of pharmacologic and genetic manipulations of head and neck squamous cell carcinoma tumor cell lines and has several advantages over other commonly used matrices. PMID- 11735817 TI - Economic evaluation of ventilation tubes in otitis media with effusion. AB - OBJECTIVE: To determine the costs and effectiveness of treatment with ventilation tubes as compared with watchful waiting in children with persistent otitis media with effusion. DESIGN: Randomized controlled trial. SETTING: Institutional practice. PATIENTS: A total of 187 young children (19 months old) with persistent bilateral otitis media with effusion. INTERVENTIONS: Treatment with ventilation tubes or watchful waiting. MAIN OUTCOME MEASURES: The time without effusion, language development, and the costs from a societal perspective during 1-year follow-up. RESULTS: The mean duration of effusion was 9.2 months in the watchful waiting group and 4.7 months in the ventilation tube group. The language development was comparable in both groups (0.7 month of improvement difference [95% confidence interval, -0.3 to 1.7 months] after correction for confounding variables). Because no statistically significant differences were found in the language development between the treatment groups, we performed a cost minimization analysis. The mean costs per child during 1 year of follow-up were 454 US dollars in the ventilation tube group and 120 US dollars in the watchful waiting group. On average, an additional investment of 334 US dollars per patient was needed for ventilation tube treatment. CONCLUSION: In the absence of differences in language development and in view of higher costs, treatment with ventilation tubes is not recommended as standard treatment in all young children with persistent otitis media with effusion traced by a population-based screening program. PMID- 11735818 TI - The safety of conscious sedation in peritonsillar abscess drainage. AB - OBJECTIVE: To demonstrate the safety of conscious sedation in draining peritonsillar abscesses (PTAs). DESIGN: Children diagnosed as having a PTA in the pediatric emergency department were identified, and their medical records were retrospectively reviewed. Results of the present study were compared with those of a previous report. SETTING: A tertiary referral children's hospital pediatric emergency department. PARTICIPANTS: Ninety-one consecutive children initially evaluated in the emergency department and managed for a PTA. INTERVENTIONS: Peritonsillar abscess incision and drainage with or without sedation. A team of physicians whose activities were documented on a formal conscious-sedation record was present. Patients were monitored for major and minor complications. OUTCOME MEASURES: The primary outcome measures were major and minor complications. Secondary outcome measures were recurrence of PTA and the need for admission. RESULTS: There were 62 episodes of conscious sedation for drainage of a PTA. Among the 91 patients, 3 had a recurrence and 24 were admitted after the procedure. A previous study evaluated 30 episodes of conscious sedation for drainage of a PTA. No major complications occurred in either series. Combining the previous data with the present data produced 92 episodes of conscious sedation for drainage of a PTA. The 1-sided upper 95% confidence limit for the rate of major complications is 3.2%. CONCLUSION: Our series, when combined with previously published data, demonstrates that conscious sedation can be safely used when draining a PTA in pediatric patients. PMID- 11735819 TI - Head and neck pilomatrixoma in children. AB - OBJECTIVE: To provide a review of the current information on the etiology, clinical presentation, management, and outcome of pilomatrixoma of the head and neck in children. DESIGN: Retrospective review. SETTING: A tertiary care pediatric center. PATIENTS: Fifty-one pediatric patients with a diagnosis of pilomatrixoma of the head and neck. INTERVENTION: All patients underwent excision of pilomatrixoma from January 1997 to March 1999. A total of 55 tumors were studied. RESULTS: A preponderance of girls (n = 36; 71%) presented with this condition. The average age at diagnosis was 5.7 years, and the average size of the lesion was 1 cm. The skin of the cheek and the periorbital area were the most commonly involved sites. Only 27 lesions (49%) had a correct preoperative diagnosis. Two (4%) of 55 tumors recurred after complete surgical excision. MAIN OUTCOME MEASURES: The age and sex of the patient, preoperative diagnosis, time elapsed before diagnosis, site and size of the tumor, length of follow-up, presence of multiple or previous pilomatrixomas, and recurrence. CONCLUSIONS: Preoperative diagnosis may be improved with increased awareness of pilomatrixoma, a common, benign skin tumor in children. Clinical findings will aid in an accurate diagnosis. Recurrence after complete local excision is rare. PMID- 11735820 TI - Congenital foregut duplication cysts of the anterior tongue. AB - OBJECTIVE: To review our experience with foregut duplication cysts of the anterior tongue, an unusual and rarely encountered mass in this location. DESIGN: A retrospective review of patients with anterior tongue foregut duplication cysts identified between 1990 and 2000. SETTING: Academic, tertiary care children's medical center. PATIENTS: Six pediatric patients (5 boys and 1 girl) ranging in age from birth to 8 months at diagnosis. INTERVENTION: Three patients underwent preoperative magnetic resonance imaging (MRI). All 6 patients underwent excisional biopsy. MAIN OUTCOME MEASURES: Clinical description of foregut duplication cysts, ability to make the diagnosis preoperatively, and recurrence rates. RESULTS: No patient presented with respiratory compromise, despite the large size of the anterior tongue masses (range, 1.5-2.4 cm). An MRI study was performed in 3 patients, all given a presumptive diagnosis of dermoid cyst based on the radiographic findings. No patient was diagnosed correctly prior to surgical excision. All patients underwent surgical excision, and the average time from birth to surgical excision was 11 months (range, 3 days to 3.7 years). Surgical pathologic findings were reported as a foregut duplication cyst (enterocystoma) in all patients, with 3 specimens containing foci of gastric mucosa. No recurrence has occurred at 1-month follow-up. CONCLUSIONS: Foregut duplication cysts rarely present in the anterior tongue and are easily misdiagnosed preoperatively. An MRI study is helpful in preoperative planning, although all lesions were radiologically indistinguishable from dermoid cysts. These masses may be an underappreciated entity in the differential diagnosis of congenital anterior tongue masses. PMID- 11735821 TI - Evidence of dysregulated cytokine production by sinus lavage and peripheral blood mononuclear cells in patients with treatment-resistant chronic rhinosinusitis. AB - BACKGROUND: Treatment-resistant chronic rhinosinusitis (CRS) imposes a clinical challenge. Its pathogenesis may be associated with dysregulated immune/inflammatory responses in the sinus. OBJECTIVE: To evaluate production of types 1 and 2 T cytokines (interferon gamma [IFN-gamma] and interleukin [IL] 5/IL 4, respectively) and regulatory/inflammatory cytokines (IL-10, IL-12, and IL-18) by sinus lavage (SL) cells and peripheral blood mononuclear cells (PBMCs) in patients with treatment-resistant CRS. METHODS: Sample SL cells and PBMCs obtained from 19 patients with treatment-resistant CRS were cultured with or without stimuli, and cytokine levels in the supernatant were measured using enzyme-linked immunosorbent assay. Control PBMC samples were obtained from 26 children. RESULTS: Chronic otitis media was found in 15 patients. Neutrophils and/or epithelial cells were dominant in SL cells. IFN-gamma, IL-12p40, and IL-10 (>100 pg/mL) were detected in SL cell cultures from 12, 9, and 8 patients, respectively. Production of IL-12p40 and IL-18 by SL cells correlated positively with phytohemagglutinin and IL-12p70 stimuli. In 12 patients, we detected IL-18 (>100 pg/mL) in SL cell cultures without stimuli, whereas PBMCs produced little IL-18, irrespective of stimuli. There was no correlation between cytokine levels produced by SL cells and PBMCs, except for IL-12p40 produced using IL-18. Decreased IFN-gamma production by PBMCs was observed in 6 patients with CRS compared with controls, but 4 of them had elevated IFN-gamma production by SL cells. Production of IL-12p40 by PBMCs was higher in 10 patients with CRS than in controls, and 7 of these patients had lower IL-10 production, resulting in an increased IL-12p40/IL-10 ratio. CONCLUSIONS: There is a role for locally produced regulatory cytokines in IFN-gamma production in the sinus in patients with treatment-resistant CRS. However, aberrant cytokine production patterns by PBMCs can be detected at high frequency in these patients, indicating that this can be used as a prognostic marker for patients with CRS. PMID- 11735822 TI - Facial sweat gland carcinoma metastasizing to neck nodes: a diagnostic and therapeutic challenge. AB - We report an unusual case involving a patient with sweat gland carcinoma of the cheek who presented with ipsilateral neck lymph node metastasis 10 years after his initial presentation. Pathological analysis of the surgical specimen revealed a strong reactivity of tumor cells to gross cystic disease fluid protein 15, estrogen receptor protein, and progesterone receptor protein. On the basis of these results, tamoxifen citrate therapy was initiated empirically. Our patient has been disease free for more than 3 years. Based on this and another case reported in the literature, we believe that antiestrogen therapy could prove beneficial in a subset of patients with sweat gland carcinoma. We recommend future multicenter clinical trials to assess the effectiveness of postoperative tamoxifen therapy for patients with estrogen and progesterone receptor protein positive metastatic sweat gland carcinoma. PMID- 11735823 TI - Eagle syndrome produced by a granular cell tumor. AB - Eagle syndrome includes elicitation of pain on swallowing, turning the head, or extending the tongue. The syndrome is thought to be caused by irritation of the glossopharyngeal nerve, most commonly caused by its impingement against an elongated styloid process. We present a rare case of a granular cell tumor presenting as Eagle syndrome. Granular cell tumors orignate from Schwann cells and are most common in the subcutaneous tissue of the head, neck, and oral cavity, especially the tongue. A granular cell tumor is typically benign and solitary, rarely malignant. The differential diagnosis, diagnostic algorithm, and treatment are presented. PMID- 11735824 TI - Pathology quiz case 1: Endolymphatic sac tumor (ELST). PMID- 11735825 TI - Pathology quiz case 2: Leiomyosarcoma of the larynx. PMID- 11735826 TI - Radiology quiz case 1: Lingual thyroid. PMID- 11735827 TI - Radiology quiz case 2: Kawasaki disease (KD) mimicking a retropharyngeal abscess. PMID- 11735828 TI - Are second-generation antihistamines appropriate for most children and adults? PMID- 11735829 TI - Antihistamines: is there really a cost-benefit controversy? PMID- 11735831 TI - The "rhinogram". PMID- 11735830 TI - Inner ear and explosions in the history of otology. PMID- 11735832 TI - Prevalence of hearing loss in a cohort of HIV-infected patients. PMID- 11735833 TI - Facial nerve functions at hospital discharge after acoustic neuroma surgery. PMID- 11735835 TI - Chromosomal aneuploidy as a predictor for poor outcome in patients with head and neck cancer. PMID- 11735838 TI - A circadian signal of change of season in patients with seasonal affective disorder. AB - BACKGROUND: In animals, the circadian pacemaker regulates seasonal changes in behavior by transmitting a signal of day length to other sites in the organism. The signal is expressed reciprocally in the duration of nocturnal melatonin secretion, which is longer in winter than in summer. We investigated whether such a signal could mediate the effects of change of season on patients with seasonal affective disorder. METHODS: The duration of melatonin secretion in constant dim light was measured in winter and in summer in 55 patients and 55 matched healthy volunteers. Levels of melatonin were measured in plasma samples that were obtained every 30 minutes for 24 hours in each season. RESULTS: Patients and volunteers responded differently to change of season. In patients, the duration of the nocturnal period of active melatonin secretion was longer in winter than in summer (9.0 +/- 1.3 vs 8.4 +/- 1.3 hours; P=.001) but in healthy volunteers there was no change (9.0 +/- 1.6 vs 8.9 +/- 1.2 hours; P=.5). CONCLUSIONS: The results show that patients with seasonal affective disorder generate a biological signal of change of season that is absent in healthy volunteers and that is similar to the signal that mammals use to regulate seasonal changes in their behavior. While not proving causality, this finding is consistent with the hypothesis that neural circuits that mediate the effects of seasonal changes in day length on mammalian behavior mediate effects of season and light treatment on seasonal affective disorder. PMID- 11735839 TI - Internal night. PMID- 11735840 TI - Focal gray matter density changes in schizophrenia. AB - BACKGROUND: The view that schizophrenia is a brain disease particularly involving decrements in gray matter is supported by findings from many imaging studies. However, it is unknown whether the (progressive) loss of tissue affects the brain globally or whether tissue loss is more prominent in some areas than in others. METHODS: Magnetic resonance whole brain images were acquired from 159 patients with schizophrenia or a schizophreniform disorder and 158 healthy subjects across a 55-year age span. Gray matter density maps were made and analyzed using voxel based morphometry. RESULTS: Compared with healthy subjects, decreases in gray matter density were found in the left amygdala; left hippocampus; right supramarginal gyrus; thalamus; (orbito) frontal, (superior) temporal, occipitotemporal, precuneate, posterior cingulate, and insular cortices bilaterally in patients with schizophrenia or schizophreniform disorder. Compared with healthy subjects, increases in gray matter density were exclusively found in the right caudate and globus pallidus in patients with schizophrenia or schizophreniform disorder. A group-by-age interaction for density was found in the left amygdala, owing to a negative regression slope of gray matter density on age in the left amygdala in patients compared with healthy subjects. CONCLUSIONS: Gray matter density is decreased in distinct focal areas in the brains of patients with schizophrenia or schizophreniform disorder. The decreased density in the left amygdala is more pronounced in older patients with schizophrenia. PMID- 11735841 TI - A randomized trial of a group cognitive intervention for preventing depression in adolescent offspring of depressed parents. AB - BACKGROUND: Adolescent offspring of depressed parents are at high risk for development of depression. Cognitive restructuring therapy holds promise for preventing progression to depressive episodes. METHODS: A randomized, controlled trial was conducted to prevent depressive episodes in at-risk offspring (aged 13 18 years) of adults treated for depression in a health maintenance organization (HMO). Potential adult cases were found by reviewing the HMO pharmacy records for dispensation of antidepressant medication and the mental health appointment system. Medical charts were reviewed for a depression diagnosis. Recruitment letters signed by treating physicians were mailed to adults. Eligible offspring had subdiagnostic depressive symptoms insufficient to meet full DSM-III-R criteria for affective disorder and/or a past mood disorder. These youth were randomized to usual HMO care (n = 49) or usual care plus a 15-session group cognitive therapy prevention program (n = 45). RESULTS: We detected significant treatment-by-time (program) effects for the Center for Epidemiological Studies Depression Scale (P=.005) and the Global Assessment of Functioning scores (P =.04). Survival analysis of incident major depressive episodes during a median 15 month follow-up found a significant advantage (P =.003) for the experimental condition (9.3% cumulative major depression incidence) compared with the usual care control condition (28.8%). CONCLUSION: A brief, group cognitive therapy prevention program can reduce the risk for depression in the adolescent offspring of parents with a history of depression. PMID- 11735842 TI - Emotional reactivity to daily life stress in psychosis. AB - BACKGROUND: The vulnerability-stress model of psychotic disorders describes, in essence, an interaction between personal vulnerability and environmental stressors. The present study investigated this interaction and studied emotional reactivity to daily life stress as a vulnerability marker for psychotic illness. METHODS: Patients with psychotic illness (n = 42), their first-degree relatives (n = 47), and control subjects (n = 49) were studied with the Experience Sampling Method (a structured diary technique assessing thoughts, current context, and mood in daily life) to assess (1) appraised subjective stress of daily events and smaller disturbances in daily life and (2) emotional reactivity conceptualized as changes in both negative affect and positive affect. RESULTS: Multilevel regression analyses showed that an increase in subjective stress was associated with an increase in negative affect and a decrease in positive affect in all groups. However, the groups differed quantitatively in their pattern of reactions to stress. Patients with psychotic illness reacted with more intense emotions to subjective appraisals of stress in daily life than control subjects. The decrease in positive affect in the relatives was similar to that of the patients, while the increase in negative affect in this group was intermediary to that of patients and control subjects. CONCLUSIONS: Higher levels of familial risk for psychosis were associated with higher levels of emotional reactivity to daily life stress in a dose-response fashion. Subtle alterations in the way persons interact with their environment may constitute part of the vulnerability for psychotic illness. PMID- 11735843 TI - Early life stress and inherited variation in monkey hippocampal volumes. AB - BACKGROUND: Opportunities for research on the causes and consequences of stress related hippocampal atrophy are limited in human psychiatric disorders. Therefore, this longitudinal study investigated early life stress and inherited variation in monkey hippocampal volumes. METHODS: Paternal half-siblings raised apart from one another by different mothers in the absence of fathers were randomized to 1 of 3 postnatal conditions that disrupted diverse aspects of early maternal care (n = 13 monkeys per condition). These conditions were previously shown to produce differences in social behavior, emotional reactivity, and neuroendocrine stress physiology. Hippocampal volumes were subsequently determined in adulthood by high-resolution magnetic resonance imaging. RESULTS: Adult hippocampal volumes did not differ with respect to the stressful postnatal conditions. Based on paternal half-sibling effects, the estimated proportion of genetic variance, ie, heritability, was 54% for hippocampal size. Paternal half siblings with small adult hippocampal volumes responded to the removal of all mothers after weaning with initially larger relative increases in cortisol levels. Plasma cortisol levels 3 and 7 days later, and measures of cortisol negative feedback in adulthood were not, however, correlated with hippocampal size. CONCLUSIONS: In humans with mood and anxiety disorders, small hippocampal volumes have been taken as evidence that excessive stress levels of cortisol induce hippocampal volume loss. Results from this study of monkeys suggest that small hippocampi also reflect an inherited characteristic of the brain. Genetically informed clinical studies should assess whether inherited variation in hippocampal morphology contributes to excessive stress levels of cortisol through diminished neuroendocrine regulation. PMID- 11735844 TI - Supervisory attentional system in nonamnesic alcoholic men. AB - BACKGROUND: Many studies have shown that recently detoxified alcoholic persons perform poorly on tasks thought to be sensitive to frontal lobe damage, supporting the hypothesis that the frontal lobes are highly vulnerable to chronic alcohol consumption. However, it appeared that most of the executive tasks used in these studies also involved nonexecutive components, and these tasks had been shown to be impaired as a result of nonfrontal lobe lesions. In this study, we examined further the "frontal lobe vulnerability" hypothesis using executive tasks, proved to be associated with frontal lobe functioning, that allowed us to distinguish the relative importance of executive and nonexecutive processes. METHOD: Thirty recently detoxified asymptomatic male alcoholic inpatients and 30 control subjects were tested for planning, inhibition, rule detection, and coordination of dual task, as well as the speed of processing and nonexecutive functions (such as short-term memory storage). RESULTS: Alcoholics performed worse than controls in almost all tasks assessing executive functions. However, they were not slower than the controls and showed normal results for nonexecutive functions. CONCLUSIONS: Chronic alcohol consumption seems to be associated with severe executive function deficits, which are still present after a protracted period of alcohol abstinence. These data support the idea that the cognitive deficits in recently detoxified sober alcoholic subjects are due, at least partly, to frontal lobe dysfunctioning. PMID- 11735845 TI - Antipsychotics and the risk of sudden cardiac death. AB - BACKGROUND: Case reports link antipsychotic drugs with sudden cardiac deaths, which is consistent with dose-related electrophysiologic effects. Because this association has not been confirmed in controlled studies, we conducted a retrospective cohort study in Tennessee Medicaid enrollees, which included many antipsychotic users; there were also computer files describing medication use and comorbidity. The study was conducted before the introduction of risperidone and, thus, did not include the newer atypical agents. METHODS: The cohort included 481,744 persons with 1,282,996 person-years of follow-up. This included 26,749 person-years for current moderate-dose antipsychotic use (>100-mg thioridazine equivalents), 31,864 person-years for current low-dose antipsychotic use, 37,881 person-years for use in the past year only, and 1 186,501 person-years for no use. The cohort had 1487 confirmed sudden cardiac deaths; from these, we calculated multivariate rate ratios adjusted for potential confounding factors. RESULTS: When current moderate-dose antipsychotic use was compared with nonuse, the multivariate rate ratio was 2.39 (95% confidence interval, 1.77-3.22; P<.001). This was greater than that for current low-dose (rate ratio, 1.30; 95% confidence interval, 0.98-1.72; P=.003) and former (rate ratio, 1.20; 95% confidence interval, 0.91-1.58; P<.001) use. Among cohort members with severe cardiovascular disease, current moderate-dose users had a 3.53-fold (95% confidence interval, 1.66-7.51) increased rate relative to comparable nonusers ( P<.001), resulting in 367 additional deaths per 10,000 person-years of follow-up. CONCLUSIONS: Patients prescribed moderate doses of antipsychotics had large relative and absolute increases in the risk of sudden cardiac death. Although the study data cannot demonstrate causality, they suggest that the potential adverse cardiac effects of antipsychotics should be considered in clinical practice, particularly for patients with cardiovascular disease. PMID- 11735846 TI - Sudden cardiac death and antipsychotic drugs: do we know enough? PMID- 11735848 TI - A thanks to the ARCHIVES community: our farewell editorial. PMID- 11735847 TI - Clozapine use in patients with schizophrenia and the risk of diabetes, hyperlipidemia, and hypertension: a claims-based approach. AB - BACKGROUND: Numerous case reports have linked clozapine to the development of diabetes mellitus and hyperlipidemia in patients with schizophrenia. However, investigators have been unable to clearly demonstrate this association when compared with a control group receiving conventional antipsychotics. METHODS: Medical and pharmacy claims from the Iowa Medicaid program were used to compare incidence rates for diabetes, hyperlipidemia, and hypertension in 552 patients receiving clozapine and 2461 patients receiving conventional antipsychotics (eg, haloperidol, chlorpromazine hydrochloride), with the use of a retrospective cohort design. Logistic regression was used to compare incidence rates adjusting for age, sex, and duration of available follow-up. RESULTS: No significant differences in overall incidence rates for diabetes, hyperlipidemia, or hypertension were observed in patients receiving clozapine vs conventional antipsychotics. However, among younger patients (aged 20-34 years), clozapine administration was associated with a significantly increased relative risk of diabetes (2.5 [95% confidence interval, 1.2-5.4]) and hyperlipidemia (2.4 [95% confidence interval, 1.1-5.2]), but not hypertension (0.9 [95% confidence interval, 0.4-2.0]). CONCLUSIONS: These data suggest that clozapine may not be an independent cause of diabetes or hyperlipidemia, but instead acts as an effect modifier in susceptible populations by increasing weight or affecting insulin secretion and resistance. This finding requires confirmation in other settings and patient populations and with the other atypical antipsychotics (risperidone, olanzapine, and quetiapine fumarate). The potential long-term medical and economic implications of the early induction of diabetes and hyperlipidemia in patients with schizophrenia warrant further study. PMID- 11735849 TI - MTA Study has flaws. PMID- 11735850 TI - MTA findings fail to consider methodological issues. PMID- 11735854 TI - Surgical textbooks and journals. PMID- 11735852 TI - Dual brain pathology in patients with affective aggressive episodes. PMID- 11735855 TI - Internationalization of general surgical journals: origin and content of articles published in North America and Great Britain from 1983 to 1998. AB - HYPOTHESIS: The origin and characteristics of articles published in the 6 highest rated (Institute for Scientific Information classification) English-language general surgical journals have changed significantly during the past 15 years. DESIGN: All articles published in 1983, 1988, 1993, and 1998 in 5 US surgical journals and 1 British surgical journal were reviewed and characterized. MAIN OUTCOME MEASURES: Absolute numbers and proportions of national and international articles published in each journal. RESULTS: Articles reviewed included 4868 in US journals and 1380 in the British journal. The total number of US journal articles decreased by 15.1%. The total number of British journal articles increased by 58.9%. The percentage of national articles decreased from 87.5% to 68.8% in US journals (P<.001) and constituted the minority of freely submitted articles in 1998 in 3 of 5 US journals. The percentage of national articles also decreased from 74.8% to 47.1% in the British journal (P<.001). Articles by European and Asian authors showed the most striking increases in all journals. The percentage of basic research articles declined in US journals from 23.3% to 17.9% (P =.001) owing to a 14.9% decline in national basic research articles. The percentage of clinical randomized studies increased from 2.2% to 4.1% (P<.008), but the increase was attributable to international articles. Government funding alone decreased from 13.6% to 11.2%, and government plus another source of funding decreased from 19.2% to 16.7% for national articles in US journals. CONCLUSIONS: Internationalization of the highly rated British and the 5 highest rated US general surgical journals has occurred. The decrease in the number of national articles in the US journals has been accompanied by significant decreases in government funding and basic research articles and a static output of clinical randomized studies from North America. PMID- 11735858 TI - The Frey procedure: local resection of pancreatic head combined with lateral pancreaticojejunostomy. AB - Management of chronic pancreatitis is mainly palliative. Most patients with chronic pancreatitis require surgical evaluation and intervention when there is suspicion of pancreatic malignancy, evidence of intractable pain, or development of pancreatitis-related local complications. The ideal operation for chronic pancreatitis, therefore, should be designed to exclude the existence of malignancy, provide long-lasting pain relief, and correct the local complications. It should be as simple and safe as possible and should preserve the remaining endocrine and exocrine functions of the pancreas. PMID- 11735859 TI - Dynamic retention: a technique for closure of the complex abdomen in critically ill patients. AB - Management of the open abdomen in the setting of massive visceral swelling or extensive intra-abdominal abscess may pose an extremely difficult surgical scenario. We herein describe the technique and results of dynamic-retention sutures used in 13 patients with abdominal catastrophes after trauma, vascular reconstruction, tumor extirpation, and intra-abdominal infection. Three of these patients died during their acute care hospitalization. The remaining 10 patients were discharged to home with no resultant fistulas and 1 recurrent hernia (10%). Dynamic-retention sutures provide a useful technique for the closure of the complex surgical abdomen. We observed a low complication rate. In properly selected patients, this technique avoids the use of mesh or additional surgical procedures such as skin grafting or plastic surgical reconstruction of the abdominal wall. PMID- 11735861 TI - Hemodynamic effects of supervised calf muscle exercise in patients with venous leg ulceration: a prospective controlled study. AB - HYPOTHESIS: Because more than two thirds of patients with venous ulcer have an impaired calf muscle pump, enhancement of its ejecting ability with physical training may generate an improved hemodynamic milieu sufficient to promoting ulcer healing. This study evaluated the effects of short-term supervised calf exercise on calf muscle pump function and venous hemodynamics in limbs with venous ulceration. DESIGN: Prospective controlled study. SETTINGS: University associated tertiary care hospital. PATIENTS: The study consisted of 2 groups. An exercise group comprised 10 patients (median age, 72 years) receiving supervised isotonic calf muscle exercise for 7 consecutive days. A control group comprised 11 patients matched with those in the exercise group for age, sex, ulcer size, and ulcer duration (all, P>.09). Patients in both groups had perimalleolar venous leg ulcers, impaired calf muscle function (ejection fraction, <60%), and full ankle joint movement. INTERVENTIONS: After providing a complete clinical history, both groups underwent a physical examination, venous duplex scanning, and air plethysmography. The venous filling index, venous volume, residual venous volume, and residual volume fraction of the calf on standing were measured plethysmographically at baseline and on day 8, in addition to calf muscle endurance as determined by the maximal number of plantar flexions performed against a fixed 4-kg resistance during 6 minutes (1 flexion/s). Operators were blinded to the subject's group. Exercise in the first group entailed consecutive active plantar flexions using a standardized 4-kg resistance pedal ergometer. Subjects daily completed 3 sets of flexions of 6 minutes each. All patients had short-stretched compression bandaging. MAIN OUTCOME MEASURES: The ejected venous volume and ejection fraction were evaluated in both groups at baseline and on day 8. RESULTS: Both groups had a similar hemodynamic performance at baseline for all the variables evaluated (P>.10). After 7 days of exercise, patients in the exercise group improved their ejected venous volume by 67.5%, ejection fraction by 62.5%, residual venous volume by 25% (all 3, P =.006), and their residual volume fraction by 28.6% (P =.008). Changes in the control group within the same period were small (all, P>.10). By day 8, the exercise group had a significantly better ejected venous volume (P<.001) and ejection fraction (P<.001) than the control group. The venous filling index and the venous volume did not change (P>.50) in either study group. Calf muscular endurance in the exercise group increased 135%, from a median 153 plantar flexions at baseline to 360 on day 7 (P<.001). CONCLUSIONS: By increasing the muscular endurance, efficacy, and power of the calf muscle, isotonic exercise improves its ejecting ability and the global hemodynamic status in limbs with venous ulceration. Prospective evaluations of the clinical effects of calf muscle pump strengthening for the treatment of venous leg ulceration are indicated by the results of this study. PMID- 11735862 TI - Preoperative cardiac risk assessment: an updated approach. AB - HYPOTHESIS: We provide an updated algorithm for approaching preoperative cardiac risk assessment in patients undergoing noncardiac surgery. DESIGN: A National Library of Medicine PubMed literature search was performed dating back to 1985 using the keywords "preoperative cardiac risk for noncardiac surgery." This search was restricted to English language articles involving human subjects. RESULTS: Patient-specific and operation-specific cardiac risk can be determined clinically. Patients with major cardiac risk factors have a high incidence of perioperative cardiac complications, whereas the risk is less than 3% for low risk patients. For intermediate-risk patients, no prospective randomized studies demonstrate the efficacy of noninvasive stress testing (dipyridamole thallium or dobutamine echocardiography) or of subsequent coronary revascularization for preventing perioperative cardiac complications. Recent studies demonstrate that perioperative beta-blockade significantly reduces the adverse cardiac event rate in intermediate-risk patients. CONCLUSIONS: Most patients with high cardiac risk should proceed with coronary angiography. Patients with low cardiac risk can proceed to surgery without noninvasive testing. For intermediate-risk patients, consideration may be given to further stress testing prior to surgery; however, in most patients, proceeding to surgery with perioperative beta-blockade is an acceptable alternative. PMID- 11735863 TI - The role of thyroid hormone administration in potential organ donors. AB - HYPOTHESIS: Levothyroxine sodium therapy should be used in brain-dead potential organ donors to reverse hemodynamic instability and to prevent cardiovascular collapse, leading to more available organs for transplantation. DESIGN: Prospective, before and after clinical study. SETTING: A surgical intensive care unit of an academic county hospital. PATIENTS: During a 12-month period (September 1, 1999, through August 31, 2000), we evaluated 19 hemodynamically unstable patients with traumatic and nontraumatic intracranial lesions, who were candidates for organ donation following brain death declaration. INTERVENTIONS: All patients were resuscitated aggressively for organ preservation by fluids, inotropic agents, and vasopressors. If, despite all measures, the patients remained hemodynamically unstable, a bolus of 1 ampule of 50% dextrose, 2 g of methylprednisolone sodium succinate, 20 U of insulin, and 20 microg of levothyroxine sodium was administered, followed by a continuous levothyroxine sodium infusion at 10 microg/h. RESULTS: There was a significant reduction in the total vasopressor requirement after levothyroxine therapy (mean +/- SD, 11.1 +/- 0.9 microg/kg per minute vs 6.4 +/- 1.4 microg/kg per minute, P =.02). Ten patients (53%) had complete discontinuation of vasopressors. There were no failures to reach organ donation due to cardiopulmonary arrest. CONCLUSIONS: Levothyroxine therapy plays an important role in the management of hemodynamically unstable potential organ donors by decreasing vasopressor requirements and preventing cardiovascular collapse. This may result in an increase in the quantity and quality of organs available for transplantation. PMID- 11735864 TI - Parathyroid autotransplantation during thyroidectomy: documentation of graft function. AB - HYPOTHESIS: Biochemical function of normal parathyroid tissue grafted during thyroidectomy can be documented. DESIGN: An intervention study in which devascularized or inadvertently removed parathyroid glands are reimplanted in forearm muscle pockets during thyroidectomy. Postoperative serum parathyroid hormone levels were evaluated by venous sampling from both forearms on postoperative days 1, 3, 14, 28, 56, and 84. SETTING: Tertiary care teaching hospital. PATIENTS: Seven patients undergoing thyroidectomy at risk for postoperative hypocalcemia. RESULTS: A 1.5-fold gradient of parathyroid hormone measurements between grafted and nongrafted arms was demonstrated in all patients on postoperative day 28. A maximal parathyroid hormone gradient was reached on day 56, and biochemical function persisted in 6 patients on day 84. CONCLUSIONS: Biochemical function of parathyroid glands reimplanted during thyroidectomy can be demonstrated objectively. The application of parathyroid autotransplantation may preserve parathyroid function for inadvertently removed or devascularized parathyroid glands during thyroid surgery. PMID- 11735865 TI - Incidence and reversibility of organ failure in the course of sterile or infected necrotizing pancreatitis. AB - BACKGROUND: Multiple organ failure (MOF) and infected necrosis are both considered severe adverse events during the course of necrotizing pancreatitis. HYPOTHESIS: The incidence of MOF and its reversibility in patients with necrotizing pancreatitis are influenced by the presence or absence of infected necrosis. DESIGN: Case series. SETTING: Intensive care, university teaching hospital. PATIENTS: Forty-three patients with necrotizing pancreatitis and failure of at least 1 organ were prospectively included. MAIN OUTCOME MEASURES: Organ failure defined according to the Goris classification; MOF defined by the simultaneous occurrence of 3 organ failures and graded with an MOF score. Microbial status of necrosis was assessed by percutaneous or intraoperative sampling. Surgical drainage was performed in patients with infected necrosis, whereas sterile necrosis was managed conservatively. RESULTS: Infected necrosis occurred in 27 patients (63%). The mean (+/-SEM) number of organ failures was greater in cases of infection (3.6 +/- 1.1 vs 2.6 +/- 1.5; P =.02). Multiple organ failure occurred more frequently in cases of infected necrosis (23/27 vs 7/16; P<.01) and was responsible for an increased mortality in this subgroup (33% vs 6%; P =.1). The severity of MOF graded by the MOF score was related to the bacteriologic status of necrosis. CONCLUSIONS: The higher mortality commonly attributed to MOF in patients with infected necrosis appears to be due to a higher frequency and an increased severity of MOF. Conservative management in patients with severe necrotizing pancreatitis and sterile necrosis complicated by MOF is supported by the high reversibility rate of MOF and the low mortality rate observed in this series. PMID- 11735866 TI - Results of a pilot trial comparing prolonged intravenous antibiotics with sequential intravenous/oral antibiotics for children with perforated appendicitis. AB - HYPOTHESIS: For children with perforated appendicitis, the use of a prolonged course of intravenous (i.v.) antibiotics is equivalent to a short course of i.v. antibiotics followed by sequential conversion to oral (PO) antibiotics. DESIGN: Prospective, randomized, clinical trial. SETTING: Multicenter study in tertiary children's hospitals. PATIENTS: Children (aged 5-18 years) with perforated appendicitis found at laparotomy. INTERVENTION: Children were randomized after appendectomy either to a 10-day course of a combination of i.v. ampicillin, gentamicin sulfate, and clindamycin (n = 10); or to a short course of a combination of i.v. ampicillin, gentamicin, and clindamycin, followed by conversion to a combination of p.o. amoxicillin and clavulanate potassium plus metronidazole (n = 16). MAIN OUTCOME MEASURES: The primary outcome measure was clinical success, which was rated as complete, partial, or failure. Secondary outcome measures included return of oral intake, duration of fever, return of normal white blood cell count, and patient charges. Treatment equivalence was determined using confidence interval analysis. RESULTS: We found treatment equivalence between the i.v. and i.v./p.o. groups, with 6 (60%) complete and 4 (40%) partial successes for the 10 patients in the i.v. group and 15 (94%) complete and 1 (6%) partial successes for the 16 patients in the i.v./p.o. group (P< or =.05). There was no difference in return of oral intake, duration of fever, or return of normal white blood cell count between the groups. Conversion to oral therapy results in savings of approximately $1500 per case. CONCLUSION: There is treatment equivalence between prolonged i.v. therapy and i.v. therapy followed by conversion to oral antibiotic therapy in children with perforated appendicitis. PMID- 11735867 TI - Surgery for ulcerative colitis in elderly persons: changes in indications for surgery and outcome over time. AB - HYPOTHESIS: Medical therapy has changed the indications for surgery over the last 4 decades. Advances in perioperative care have significantly improved the outcome. DESIGN: The medical records of all patients 65 years and older who underwent surgery for ulcerative colitis during a 40-year period were analyzed retrospectively. SETTING: Tertiary referral center. PATIENTS: One hundred thirteen consecutive patients 65 years and older who underwent surgery for ulcerative colitis between January 1, 1960, and June 30, 1999. MAIN OUTCOME MEASURES: Changes in elective and urgent indications for surgery. Changes over time in outcome and the factors that brought about these changes. Predictors of poor outcome in an elderly population with ulcerative colitis. RESULTS: One hundred thirteen patients were divided into 3 cohorts of 38, 38, and 37 consecutive patients admitted to the hospital during the periods 1960 through 1984, 1985 through 1993, and 1994 through 1999, respectively. Indications for surgery and morbidity and mortality rates have changed with time. Dysplasia has replaced carcinoma as a major indication for elective surgery (P =.001). Toxic megacolon has become significantly less common as an indication for urgent surgery (P =.001). Surgery-associated adverse outcomes have decreased significantly from 50% (13% deaths, 37% major complications) to 27% (3% deaths, 24% major complications) (P =.04). Male sex, an albumin level of 2.8 g/dL or less, and urgent surgery were found to be independent predictors of poor outcome. CONCLUSIONS: In our referral center, the indications for urgent and elective surgery have changed during the past 4 decades from toxic megacolon and carcinoma, to disease refractory, to medical therapy and dysplasia, respectively. Morbidity and mortality have decreased dramatically over time. Urgent procedures, low levels of albumin, and male sex are all predictors of poor outcome. PMID- 11735868 TI - Temporal assessment of Candida risk factors in the surgical intensive care unit. AB - HYPOTHESIS: Risk factors for Candida infection in surgical intensive care units (SICUs) change over time. Risk factor progression may influence Candida colonization and infection. DESIGN: Multicenter cohort survey. SETTING: Three urban teaching institutions. PATIENTS: A total of 301 consecutively admitted patients in SICUs for 5 or more days. MAIN OUTCOME MEASURES: Assessment of patients on SICU days 1, 3, 4, 6, and 8 and SICU discharge for risk factors, Candida colonization, and antifungal use. Candida colonization status was categorized as noncolonized (NC), locally colonized (LC) if 1 site was involved, and disseminated infection (DI) if 2 or more sites or candidemia were involved. RESULTS: The most frequent risk factors in the 301 patients enrolled were presence of peripheral and central intravenous catheters, bladder catheters, mechanical ventilation, and lack of enteral or intravenous nutrition. Early risk factors included total parenteral nutrition or central catheter at SICU day 1 and previous SICU admissions or surgical procedures. Peak number of risk factors (mean +/- SD) were as follows: 7.2 +/- 2.6 in NC (n = 229), 9.2 +/- 2.3 in LC (n = 45), and 9.2 +/- 2.6 in DI (n = 27). These numbers were reached at day 8 in the NC and LC groups and day 4 in the DI group. The LC and DI groups had more risk factors on each SICU day than the NC group and longer median SICU length of stay (28 days in the DI group vs 11 and 19 days in the NC and LC groups, respectively). Antifungal therapy, while used most frequently in the DI group, was initiated later for this group than in NC and LC groups. CONCLUSIONS: Risk factors for Candida infection in SICU patients change over time. Patients with DI demonstrate a greater number of and more rapid increase in risk factors than patients in the LC and NC groups. Presence of early risk factors at the time of SICU admission, a high incidence of risk factors, or a rapid increase in risk factors should prompt clinicians to obtain surveillance fungal cultures and consider empirical antifungal therapy. PMID- 11735870 TI - Clinical features and management of hepatic portal venous gas: four case reports and cumulative review of the literature. AB - HYPOTHESIS: Hepatic portal venous gas (HPVG) has been considered a rare entity associated with a grave prognosis. Since 1978, when Liebman et al reviewed 64 cases of HPVG and reported a mortality of 75%, the number of reported cases has been increasing. DESIGN: Case series. PATIENTS AND METHODS: We reviewed the literature on 182 cases of HPVG in adults, including 4 of our patients, (transplantation and abdominal trauma cases were excluded) and analyzed the cause, pathogenesis, and clinical features. RESULTS: In this series, the underlying clinical events associated with HPVG were bowel necrosis (43%), digestive tract dilatation (12%), intraperitoneal abscess (11%), ulcerative colitis (4%), gastric ulcer (4%), Crohn disease (4%), complications of endoscopic procedures (4%), intraperitoneal tumor (3%), and other (15%). The overall mortality was 39% but varied depending on the underlying disease. CONCLUSIONS: Hepatic portal venous gas is a lethal or curable entity caused by various diseases. The underlying disease associated with HPVG determines the clinical features and prognosis of the patients. The treatment of patients with HPVG should be directed to the underlying disease. PMID- 11735871 TI - Surgery in the "New" Hong Kong. PMID- 11735872 TI - Human immunodeficiency virus and malignancy: thoughts on viral oncogenesis. PMID- 11735873 TI - Resident work hours: the evolution of a revolution. PMID- 11735875 TI - Blue urticaria: a previously unreported adverse event associated with isosulfan blue. AB - The sentinel node hypothesis is predicated on the fact that a metastasis, if it exists, will have traveled on a direct path from the primary tumor through the efferent lymphatic channels to the first draining lymph node in the regional lymphatic basin, the sentinel node. Lymphatic mapping with isosulfan blue and sentinel lymphadenectomy is being increasingly used in the management of patients with melanoma, breast cancer, and other solid tumors. This trend is exposing an increasing number of patients to isosulfan blue. Although this compound is generally safe, severe reactions have been reported. We describe 2 patients who developed "blue hives" after isosulfan blue injection. PMID- 11735876 TI - Image of the month. Acute gastric dilation with necrosis. PMID- 11735877 TI - Surgical reminiscences: teaching and learning evidence-based surgical practice: a tale of 3 Texans. PMID- 11735878 TI - Valentine Mott and the beginnings of vascular surgery. PMID- 11735879 TI - Domain coarsening of stripe patterns close to onset. AB - We study domain coarsening of two-dimensional stripe patterns by numerically solving the Swift-Hohenberg model of Rayleigh-Benard convection. Near the bifurcation threshold, the evolution of disordered configurations is dominated by grain-boundary motion through a background of largely immobile curved stripes. A numerical study of the distribution of local stripe curvatures, of the structure factor of the order parameter, and a finite size scaling analysis of the grain boundary perimeter, suggest that the linear scale of the structure grows as a power law of time t(1/z), with z=3. We interpret theoretically the exponent z=3 from the law of grain-boundary motion. PMID- 11735880 TI - Time scales in rotating unstable Langevin-type dynamics. AB - In this Rapid Communication we propose a different and general characterization of rotating, unstable Langevin-type dynamics in the presence of an external force in the context of two dynamical representations x and y, using the passage time distribution. Here y is the transformed space of coordinates obtained by means of a time-dependent rotation matrix. The Langevin dynamics in the new y space defines an interesting concept of external force and internal noise due to rotation. The theory is applied to the characterization of rotational unstable systems of two (such as the laser system) and three variables, and stimulates its application in other fields, for instance, in plasma physics. PMID- 11735881 TI - Onset condition of modulated Rayleigh-Benard convection at low frequency. AB - The onset condition of convection in a layer of fluid bounded by isothermal walls, with lower temperature varying sinusoidally in time at very low nondimensional modulation frequency, is derived in closed form, based on the Floquet theory and using a matched-asymptotic WKB method. PMID- 11735882 TI - Velocity correlations in dense granular gases. AB - We report the statistical properties of spherical steel particles rolling on an inclined surface being driven by an oscillating wall. Strong dissipation occurs due to collisions between the particles and can be tuned by changing the number density. The velocities of the particles are observed to be correlated over large distances comparable to the system size. The distribution of velocities deviates strongly from a Gaussian. The degree of the deviation, as measured by the kurtosis of the distribution, is observed to be as much as four times the value corresponding to a Gaussian, signaling a significant breakdown of the assumption of negligible velocity correlations in a granular system. PMID- 11735883 TI - Pair interaction of charged colloidal spheres near a charged wall. AB - Although equally charged colloidal particles dispersed in clean water are expected to repel each other, an unexplained long-range attraction has consistently been reported for charged colloidal spheres confined by charged macroscopic surfaces. We present an alternative equilibrium measurement of the pair interaction energy for charged spheres near a single charged wall. Analyzing their radial distribution functions for different concentrations reveals a purely repulsive sphere-sphere interaction that is well described by a screened Coulomb potential. PMID- 11735884 TI - Singular thin viscous sheet. AB - The evolution of a thin viscous layer is usually smooth. Here we conduct an experiment where the layer adopts a singular shape. Using the analogy between the flow of a viscous liquid and the deformation of an elastic solid, the theoretical analysis predicts a conical shape for the sheet and is in quantitative agreement with the experiment. PMID- 11735885 TI - Compositionally modulated phase in crystalline n-alkane mixtures. AB - We report an x-ray scattering study of the compositionally modulated microphase separated state (muPSS) of binary n-alkane (C23H48:C28H58) mixtures. By employing a quenching technique, we obtained many orders of sharp lamellar and superlattice reflections. The muPSS is a regular superstructure consisting of pure C23 layers, and layers rich in C28 but containing up to approximately 14% C23 plus voids. A monolayer of the minority component regularly alternates with one or more layers of the majority. The mechanism for thermodynamic stability of the muPSS may be related to the entropy associated with longitudinal displacements of the molecules. PMID- 11735886 TI - Orientational ordering in the chiral smectic-C*FI2 liquid crystal phase determined by resonant polarized x-ray diffraction. AB - High-resolution resonant polarized x-ray diffraction experiments near the sulfur K edge have been performed on free-standing liquid crystal films exhibiting the chiral smectic-C*FI2 phase. It is widely accepted that this phase has a four layer repeat unit, but the internal structure of the repeat unit remains controversial. We report different resolved features of the resonant x-ray diffraction peaks associated with the smectic-C*FI2 phase that unambiguously demonstrate that the four-layer repeat unit is locally biaxial about the layer normal and that the measured angle, describing the biaxiality, is in good agreement with optical measurements. PMID- 11735887 TI - Direct layer-by-layer freezing of a smectic liquid-crystal surface into the crystalline phase. AB - Electron diffraction and optical reflectivity have provided the direct confirmation of the existence of layer-by-layer surface transitions from the smectic-A immediately to the crystal-B phase in a liquid-crystal material, without going through an intermediate hexatic phase. The molecular interactions are found to be through retarded van der Waals forces. Our results suggest that a smectic-A film can transform into a crystal-B through three possible scenarios. PMID- 11735888 TI - Bethe approximation for a model of polymer solvation. AB - The phase diagram of a recently proposed model for the solvation of monomers and polymers in water is studied in the homopolymer case, and several thermodynamic quantities are computed by means of pair approximation of the cluster variation method, i.e., the Bethe approximation. The model takes into account the water degrees of freedom in a simplified way, so that they can be integrated out analytically. The resulting effective Hamiltonian contains only a temperature dependent water-monomer interaction and its phase diagram can be easily studied thanks to the simplicity of the Bethe approximation and exhibits, for a hydrophobic polymer, both cold and warm unfolding transitions in a wide region of the parameter space. This suggests that the present one might be a toy-model description of the phase behavior observed experimentally in water solutions of hydrophobic polymers, such as poly-N-isopropylacrylamide (PNIPAM), as well as a step to understand the mechanism of cold unfolding in proteins. PMID- 11735889 TI - Surface-tethered chains entangled in a polymer melt: effects on adhesion dynamics. AB - The adhesion between a polymer melt and substrate is studied in the presence of chemically attached chains on the substrate surface. Extensive molecular dynamics simulations have been carried out to study the effect of temperature, tethered chain density (Sigma), tethered chain length (N(t)), and tensile pull velocity (v) on the adhesive failure mechanisms of pullout and/or scission of the tethered chains. We observe a crossover from pure chain pullout to chain scission as N(t) is increased. The value of N(t) for which this crossover begins approaches the bulk entanglement length N(e) as the temperature is lowered. PMID- 11735890 TI - Short- and intermediate-time behavior of the linear stress relaxation in semiflexible polymers. AB - The linear viscoelasticity of semiflexible polymers is studied through Brownian Dynamics simulations covering a broad range of chain stiffness and time scales. Our results agree with existing theoretical predictions in the flexible and stiff limits; however, we find that over a wide intermediate-time window spanning several decades, the stress relaxation is described by a single power law t( alpha), with the exponent alpha apparently varying continuously from 1/2 for flexible chains, to 5/4 for stiff ones. Our study identifies the limits of validity of the t(-3/4) power law at short times predicted by recent theories. An additional regime is identified, the "ultrastiff" chains, where this behavior disappears. In the absence of Brownian motion, the purely mechanical stress relaxation produces a t(-3/4) power law for both short and intermediate times. PMID- 11735891 TI - Critical behavior of propagation on small-world networks. AB - We report numerical evidence that an epidemiclike model, which can be interpreted as the propagation of a rumor, exhibits critical behavior at a finite randomness of the underlying small-world network. The transition occurs between a regime where the rumor "dies" in a small neighborhood of its origin, and a regime where it spreads over a finite fraction of the whole population. Critical exponents are evaluated through finite-size scaling analysis, and the dependence of the critical randomness with the network connectivity is studied. The behavior of this system as a function of the small-network randomness bears noticeable similarities with an epidemiological model reported recently [M. Kuperman and G. Abramson, Phys. Rev. Lett. 86, 2909 (2001)], in spite of substantial differences in the respective dynamical rules. PMID- 11735892 TI - Random dynamics of the Hodgkin-Huxley neuron model. AB - Noise can alter the response of neurons, enhancing their ability to detect weak inputs. We analyze how the Hodgkin-Huxley equations, a canonical neuron model, respond to white noise stimulation. We show that this model possesses a stochastic attractor, reduced to a unique stochastic equilibrium point that attracts all trajectories. PMID- 11735893 TI - Instability of myelin tubes under dehydration: deswelling of layered cylindrical structures. AB - We report experimental observations of an undulational instability of myelin figures. Motivated by this, we examine theoretically the deformation and possible instability of concentric, cylindrical, multilamellar membrane structures. Under conditions of osmotic stress (swelling or dehydration), we find a stable, deformed state in which the layer deformation is given by deltaR infinity r(square root[B(A)/(hB)]), where B(A) is the area compression modulus, B is the interlayer compression modulus, and h is the repeat distance of layers. Also, above a finite threshold of dehydration (or osmotic stress), we find that the system becomes unstable to undulations, first with a characteristic wavelength of order square root[xi(d)0], where xi is the standard smectic penetration depth and d0 is the thickness of dehydrated region. PMID- 11735894 TI - Slow crossover to Kardar-Parisi-Zhang scaling. AB - The Kardar-Parisi-Zhang (KPZ) equation is accepted as a generic description of interfacial growth. In several recent studies, however, values of the roughness exponent alpha have been reported that are significantly less than that associated with the KPZ equation. A feature common to these studies is the presence of holes (bubbles and overhangs) in the bulk and an interface that is smeared out. We study a model of this type in which the density of the bulk and sharpness of the interface can be adjusted by a single parameter. Through theoretical considerations and the study of a simplified model we determine that the presence of holes does not affect the asymptotic KPZ scaling. Moreover, based on our numerics, we propose a simple form for the crossover to the KPZ regime. PMID- 11735895 TI - Exact multilocal renormalization group and applications to disordered problems. AB - We develop a method, the exact multilocal renormalization group (EMRG) which applies to a broad set of theories. It is based on the systematic multilocal expansion of the Polchinski-Wilson exact renormalization group (ERG) equation together with a scheme to compute correlation functions. Integrating out explicitly the nonlocal interactions, we reduce the ERG equation obeyed by the full interaction functional to a flow equation for a function, its local part. This is done perturbatively around fixed points, but exactly to any given order in the local part. It is thus controlled, at variance with projection methods, e.g., derivative expansions or local potential approximations. Our EMRG method is well-suited to problems such as the pinning of disordered elastic systems, previously described via functional renormalization group (FRG) approach based on a hard cutoff scheme. Since it involves arbitrary cutoff functions, we explicitly verify universality to O(epsilon=4-D), both of the T=0 FRG equation and of correlations. Extension to finite temperature T yields the finite size (L) susceptibility fluctuations characterizing mesoscopic behavior (Deltachi)2 approximately L(straight theta)/T, where straight theta is the energy exponent. Finally, we obtain the universal scaling function to O(epsilon(1/3)) which describes the ground state of a domain wall in a random field confined by a field gradient, compare with exact results and variational method. Explicit two loop exact RG equations are derived and the application to the FRG problem is sketched. PMID- 11735896 TI - Origin of the hydrodynamic Lyapunov modes. AB - Recent studies of the Lyapunov spectrum of the hard sphere fluid reveal that there are "hydrodynamic" Lyapunov exponents corresponding to collective perturbations in phase space. We show that these collective perturbations are due to the conservation of certain quantities during collisions. These new conservation laws generate new hydrodynamic fields, just as the conservation of mass, momentum, and energy generate the density, velocity, and temperature fields. We then construct a detailed theory of the new hydrodynamic fields using a kinetic theory approach. This theory predicts several properties of the modes, but not all of them. This suggests that the underlying idea is correct, but a detailed theory must be elaborated in another way. The hydrodynamic exponents are not related in a simple way to the transport coefficients. PMID- 11735897 TI - Estimating and improving the signal-to-noise ratio of time series by symbolic dynamics. AB - We investigate the effect of symbolic encoding applied to times series consisting of some deterministic signal and additive noise, as well as time series given by a deterministic signal with randomly distributed initial conditions as a model of event-related brain potentials. We introduce an estimator of the signal-to-noise ratio (SNR) of the system by means of time averages of running complexity measures such as Shannon and Renyi entropies, and prove its asymptotical equivalence with the linear SNR in the case of Shannon entropies of symbol distributions. A SNR improvement factor is defined, exhibiting a maximum for intermediate values of noise amplitude in analogy to stochastic resonance phenomena. We demonstrate that the maximum of the SNR improvement factor can be shifted toward smaller noise amplitudes by using higher order Renyi entropies instead of the Shannon entropy. For a further improvement of the SNR, a half wave encoding of noisy time series is introduced. Finally, we discuss the effect of noisy phases on the linear SNR as well as on the SNR defined by symbolic dynamics. It is shown that longer symbol sequences yield an improvement of the latter. PMID- 11735898 TI - Experimental evidence of stochastic resonance without tuning due to non-Gaussian noises. AB - In order to test theoretical predictions, we have studied the phenomenon of stochastic resonance in an electronic experimental system driven by white non Gaussian noise. In agreement with the theoretical predictions our main findings are an enhancement of the sensibility of the system together with a remarkable widening of the response (robustness). This implies that even a single resonant unit can reach a marked reduction in the need for noise tuning. PMID- 11735899 TI - Fractional Langevin equation. AB - We investigate fractional Brownian motion with a microscopic random-matrix model and introduce a fractional Langevin equation. We use the latter to study both subdiffusion and superdiffusion of a free particle coupled to a fractal heat bath. We further compare fractional Brownian motion with the fractal time process. The respective mean-square displacements of these two forms of anomalous diffusion exhibit the same power-law behavior. Here we show that their lowest moments are actually all identical, except the second moment of the velocity. This provides a simple criterion that enable us to distinguish these two non Markovian processes. PMID- 11735900 TI - Noise-induced phase synchronization enhanced by dichotomic noise. AB - We study the nonlinear response of a stochastic bistable system driven by both a weak periodic signal and a dichotomic noise in terms of stochastic phase synchronization. We show that the effect of noise-induced phase synchronization can be significantly enhanced by the addition of a dichotomic noise. PMID- 11735901 TI - Anomalous two-state model for anomalous diffusion. AB - An anomalous two-state model (ATSM) with the anomalous long-tailed kinetics of transitions between states is proposed to describe the specific features of anomalous diffusion (AD) and AD-assisted transitions (ADAT) in the double-well potential. In the ATSM the system is assumed to undergo the conventional diffusion in both states but with different diffusion coefficients. The anomalous features of diffusion result from the modulation of the diffusion coefficient caused by transitions between ATSM states. The anomalous space-time evolution predicted by the ATSM is treated within the continuous time random walk theory. With the use of the proposed ATSM the transient behavior of the AD and the ADAT is analyzed in detail. We found a large variety of different (and sometimes peculiar) types of the space-time behavior of the free AD and ADAT. The free AD is found to be of subdiffusion or superdiffusion type for fairly long time depending on the relation between the parameters of the ATSM. The kinetics of the ADAT can be either conventional (exponential) or anomalous (of inverse power type) for different parameters of the model and time. PMID- 11735902 TI - Effect of external noise correlation in optical coherence resonance. AB - Coherence resonance occurring in semiconductor lasers with optical feedback is studied via the Lang-Kobayashi model with external nonwhite noise in the pumping current. The temporal correlation and the amplitude of the noise have a highly relevant influence in the system, leading to an optimal coherent response for suitable values of both the noise amplitude and correlation time. This phenomenon is quantitatively characterized by means of several statistical measures. PMID- 11735903 TI - Robust stochastic resonance: signal detection and adaptation in impulsive noise. AB - Stochastic resonance (SR) occurs when noise improves a system performance measure such as a spectral signal-to-noise ratio or a cross-correlation measure. All SR studies have assumed that the forcing noise has finite variance. Most have further assumed that the noise is Gaussian. We show that SR still occurs for the more general case of impulsive or infinite-variance noise. The SR effect fades as the noise grows more impulsive. We study this fading effect on the family of symmetric alpha-stable bell curves that includes the Gaussian bell curve as a special case. These bell curves have thicker tails as the parameter alpha falls from 2 (the Gaussian case) to 1 (the Cauchy case) to even lower values. Thicker tails create more frequent and more violent noise impulses. The main feedback and feedforward models in the SR literature show this fading SR effect for periodic forcing signals when we plot either the signal-to-noise ratio or a signal correlation measure against the dispersion of the alpha-stable noise. Linear regression shows that an exponential law gamma(opt)(alpha)=cA(alpha) describes this relation between the impulsive index alpha and the SR-optimal noise dispersion gamma(opt). The results show that SR is robust against noise "outliers." So SR may be more widespread in nature than previously believed. Such robustness also favors the use of SR in engineering systems. We further show that an adaptive system can learn the optimal noise dispersion for two standard SR models (the quartic bistable model and the FitzHugh-Nagumo neuron model) for the signal-to-noise ratio performance measure. This also favors practical applications of SR and suggests that evolution may have tuned the noise-sensitive parameters of biological systems. PMID- 11735904 TI - Accurate estimation of the survival probability for trapping in two dimensions. AB - In this work we study the mean survival probability Phi(n,c) of random walks on a two-dimensional lattice in the presence of traps of concentration c, as a function of the number of steps n. The computation of this quantity is performed indirectly by using the distribution of the number of sites visited S(n). In order to achieve an accurate description of this distribution we use a combination of numerical techniques. The method allows an accurate calculation of Phi down to very small values (of the order of 10(-100), for example), which is not possible via direct simulations. The survival probability is analyzed in terms of an asymptotic expansion, following the results of Donsker and Varadhan [Commun. Pure Appl. Math. 28, 525 (1975); 32, 721 (1979)], and by using the outcome of a scaling ansatz, as described in our earlier work. PMID- 11735905 TI - Self-generated disorder and structural glass formation in homopolymer globules. AB - We have investigated the interrelation between spin glasses and structural glasses. Spin glasses in this case are p-spin interaction spin glasses (at p>2) or Potts glasses that contain quenched disorder, whereas the structural glasses are here exemplified by a homopolymeric globule, which can be viewed as a liquid of connected units on a nanoscale. It is argued that the homopolymeric globule problem can be mapped onto a disorder field theoretical model whose effective Hamiltonian resembles the corresponding one for the spin glass model. In this sense the disorder in the globule is self-generated (in contrast to spin glasses) and can be related to competing interactions (virial coefficients of different signs) and the chain connectivity. The work is aimed at giving a quantitative description of this analogy. We have investigated in the mean-field approximation the phase diagram of the homopolymeric globule where the transition line from the liquid to glassy globule is treated in terms of the replica symmetry breaking paradigm. The configurational entropy temperature dependence is also discussed. PMID- 11735906 TI - Rotational Brownian motion of axisymmetric particles in a Maxwell fluid. AB - A theory of non-Markovian rotational Brownian motion is developed for axisymmetric particles moving in a Maxwell fluid in the presence of an external field. Both the inertial and viscoelastic effects are taken into account. A kinetic equation for the joint probability distribution of orientation, angular velocity, and acceleration of a particle without spin is derived starting from the rotational Langevin equation with relaxed hydrodynamic and random torques. A third-order stochastic differential equation for the particle orientation vector is also derived. Directly from this equation, the set of nonlinear evolution equations for one-time moments is derived in a noninertial approximation. The expressions for a linear response to a time-dependent external field and dynamic susceptibility of particle are obtained by direct averaging of particle orientation equation. Appendices derive the rotational mobility of axisymmetric particles in a general linear viscoelastic fluid, and the evolution equations for one-time moments of the orientation vector for axisymmetric particles moving in a Maxwell fluid in the presence of an external field. PMID- 11735907 TI - Correlation ratchets: four current reversals and disjunct "windows". AB - Multinoise correlation ratchets with a simple sawtooth potential are considered. It is proved that in the case of symmetric nonequilibrium three-level Markovian noise the direction and value of the induced current can be controlled by thermal noise. Moreover, it is established that four current reversals (CRs) occur and that for the CRs there exist characteristic disjunct "windows" in temperature and switching rate as control parameters. The necessary and sufficient conditions for the existence of the above effects are given and can be used in particle separation techniques. PMID- 11735908 TI - Amphiphilic hard body mixtures. AB - In order to study ternary amphiphilic mixtures, we introduce a simplistic model of hard spheres corresponding to water and hard needles corresponding to oil and amphiphilic particles, where the hydrophilic head is modeled as a hard sphere and the hydrophobic tail as an infinitely thin needle attached radially to the sphere. For this system, we construct a geometry-based density functional and perform Monte Carlo computer simulations. The equation of state derived from the theory is found to be in remarkable agreement with our simulation results. We investigate the theoretical demixing phase diagram, and find that the predicted trends strongly support the amphiphilic character of the model. PMID- 11735909 TI - Stochastic resonance in delayed two-coupled oscillators without common perturbations. AB - Stochastic resonance (SR) in a two-coupled oscillator system with time delay is investigated. The system shows multistability of a desynchronized state and two synchronized states with different collective frequencies, which may be interpreted as multistable perception of ambiguous or reversible figures. To model the situations where the two oscillators exist in different environments, periodic signals and noises at their inputs are not uniformly given. SR in an individual oscillator, characterized by the output signal-to-noise ratio, is examined based on numerical simulations. We find that phase shift between the signals at inputs of different oscillators weakens SR, the oscillator with only an input signal does not show SR, and the oscillator with input noise shows SR irrespective of it having an input signal or not. The results have implications in the area of information transmission in biological systems. PMID- 11735910 TI - Finite-size effects in molecular dynamics simulations: intermediate scattering function and velocity of sound. III. Theory and application to a model krypton fluid. AB - We describe a method for obtaining the intermediate scattering function I(Q,t) from a computer simulation: it is an extension of our earlier calculation [Salacuse, Denton, and Egelstaff, Phys. Rev. E 53, 2382 (1996)] for the t-->0 limit. We use this approach to obtain I(Q,t) for low Q and t from molecular dynamics (MD) simulations of a model krypton fluid whose atoms interact via a truncated Aziz pair potential, and the results are compared over their range of validity to I(Q,t) determined by the standard MD method and also by a time expansion approach. In its range of validity our approach is much more efficient than the standard MD method; however, it covers a restricted range of t due to the movement of density fluctuations (sound waves) through the simulated fluid which produces an anomaly in the time behavior of I(Q,t). By analyzing I(Q=0,t) the velocity of sound in the simulation is determined, and the results compare favorably with published experimental results for the sound velocity of liquid krypton. PMID- 11735911 TI - Theory of asymmetric nonadditive binary hard-sphere mixtures. AB - It is shown that the formal procedure of integrating out the degrees of freedom of the small spheres in a binary hard-sphere mixture works equally well for nonadditive as it does for additive mixtures. For highly asymmetric mixtures (small size ratios) the resulting effective Hamiltonian of the one-component fluid of big spheres, which consists of an infinite number of many-body interactions, should be accurately approximated by truncating after the term describing the effective pair interaction. Using a density functional treatment developed originally for additive hard-sphere mixtures the zero, one, and two body contribution to the effective Hamiltonian are determined. It is demonstrated that even small degrees of positive or negative nonadditivity have significant effect on the shape of the depletion potential. The second virial coefficient B2, corresponding to the effective pair interaction between two big spheres, is found to be a sensitive measure of the effects of nonadditivity. The variation of B2 with the density of the small spheres shows significantly different behavior for additive, slightly positive and slightly negative nonadditive mixtures. Possible repercussions of these results for the phase behavior of binary hard-sphere mixtures are discussed and it is suggested that measurements of B2 might provide a means of determining the degree of nonadditivity in real colloidal mixtures. PMID- 11735912 TI - Angle of repose and segregation in cohesive granular matter. AB - We study the effect of fluids on the angle of repose and the segregation of granular matter poured into a silo. The experiments are conducted in two regimes where: (i) the volume fraction of the fluid (liquid) is small and it forms liquid bridges between particles thus giving rise to cohesive forces, and (ii) the particles are completely immersed in the fluid. The data is obtained by imaging the pile formed inside a quasi-two-dimensional silo through the transparent glass side walls and using color-coded particles. In the first series of experiments, the angle of repose is observed to increase sharply with the volume fraction of the fluid and then saturates at a value that depends on the size of the particles. We systematically study the effect of viscosity by using water glycerol mixtures to vary it over at least three orders of magnitude while keeping the surface tension almost constant. Besides surface tension, the viscosity of the fluid is observed to have an effect on the angle of repose and the extent of segregation. In case of bidisperse particles, segregation is observed to decrease and finally saturate depending on the size ratio of the particles and the viscosity of the fluid. The sharp initial change and the subsequent saturation in the extent of segregation and angle of repose occurs over similar volume fraction of the fluid. Preferential clumping of small particles causes layering to occur when the size of the clumps of small particles exceeds the size of large particles. We calculate the azimuthal correlation function of particle density inside the pile to characterize the extent of layering. In the second series of experiments, particles are poured into a container filled with a fluid. Although the angle of repose is observed to be unchanged, segregation is observed to decrease with an increase in the viscosity of the fluid. The viscosity at which segregation decreases to zero depends on the size ratio of the particles. PMID- 11735913 TI - Granular flow down an inclined plane: Bagnold scaling and rheology. AB - We have performed a systematic, large-scale simulation study of granular media in two and three dimensions, investigating the rheology of cohesionless granular particles in inclined plane geometries, i.e., chute flows. We find that over a wide range of parameter space of interaction coefficients and inclination angles, a steady-state flow regime exists in which the energy input from gravity balances that dissipated from friction and inelastic collisions. In this regime, the bulk packing fraction (away from the top free surface and the bottom plate boundary) remains constant as a function of depth z, of the pile. The velocity profile in the direction of flow vx(z) scales with height of the pile H, according to vx(z) proportional to H(alpha), with alpha=1.52+/-0.05. However, the behavior of the normal stresses indicates that existing simple theories of granular flow do not capture all of the features evidenced in the simulations. PMID- 11735914 TI - Modeling relaxation and jamming in granular media. AB - We introduce a stochastic microscopic model to investigate the jamming and reorganization of grains induced by an object moving through a granular medium. The model reproduces the experimentally observed periodic sawtooth fluctuations in the jamming force and predicts the period and the power spectrum in terms of the controllable physical parameters. It also predicts that the avalanche sizes, defined as the number of displaced grains during a single advance of the object, follow a power law P(s) approximately s(-tau), where the exponent is independent of the physical parameters. PMID- 11735915 TI - Numerical solution of the Smoluchowski equation for a vibrofluidized granular bed. AB - A stochastic approach, similar to that used to describe Brownian motion, was used to model the displacement probability of grains in a three-dimensional vibrofluidized granular bed. As neither an analytical description nor measurements of the diffusion coefficients were available, the governing partial differential equation, namely, the Smoluchowski equation, was solved numerically using an iterative procedure, modifying the granular temperature profile at each step. The results of this stochastic model were compared to experimental measurements of the displacement probability density made using positron emission particle tracking. The results indicate that methods based on hard elastic systems such as the Smoluchowski equation are appropriate to granular systems, particularly over timescales greater than the mean collision time. PMID- 11735916 TI - Nonequilibrium phase transition for a heavy particle in a granular fluid. AB - It is shown that the homogeneous cooling state (HCS) for a heavy impurity particle in a granular fluid supports two distinct phases. The order parameter straight phi;(s) is the mean square velocity of the impurity particle relative to that of a fluid particle, and the control parameter xi* is the fluid cooling rate relative to the impurity collision rate. For xi*<1 there is a "normal" phase for which straight phi;(s) scales as the fluid/impurity mass ratio, just as for a system with elastic collisions. For xi*>1 an "ordered" phase occurs in which straight phi;(s) is finite even for vanishingly small mass ratio, representing an extreme violation of energy equipartition. The phenomenon can be described in terms of a Landau-like free energy for a second order phase transition. The dynamics leading to the HCS is studied in detail using an asymptotic analysis of the Enskog-Lorentz kinetic equation near each phase and the critical domain. Critical slowing is observed with a divergent relaxation time at the critical point. The stationary velocity distributions are determined in each case, showing a crossover from Maxwellian in the normal phase to an exponential quartic function of the velocity that is sharply peaked about the nonzero straight phi;(s) for the ordered phase. It is shown that the diffusion coefficient in the normal phase diverges at the critical point and remains so in the ordered phase. This is interpreted as a transition from diffusive to ballistic dynamics between the normal and ordered phases. PMID- 11735917 TI - Buckling and nonlocal elasticity of charged membranes. AB - The elastic behavior of an interacting, and, in specific, of a charged flexible membrane is considered. In the first part of this paper the effective nonlocal elastic energy of a membrane due to a pairwise and arbitrary intra-membrane interaction is derived. Nonlocal elasticity is included to all orders, this description, therefore, corresponds to an infinite resummation of the standard gradient expansion. In the second part, the pair interaction between segments of an (on average) neutral membrane consisting of mobile positive and negative charges is derived both field theoretically in the Gaussian approximation and using a simple ion-pairing approximation. This model might also apply to strongly charged membranes with strongly condensed counter ions. The resulting contribution to the elastic energy is negative and thus favors undulations of the membrane. The bending modulus is extracted from the large-scale or small-momentum behavior of the elastic kernel and found to be comparable to k(B)T for the case where ion pairing is dominant. The large-momentum elastic response exhibits a markedly different scaling than the small-momentum regime and sensitively depends on the small-distance cutoff and thus on molecular details. PMID- 11735918 TI - Dipolar-interaction-induced fractal pattern formation in magnetic multilayers. AB - Models concerning particle diffusion and aggregation have been proposed for decades, and the aggregations with long-range dipolar interaction are simulated and analyzed numerically. In this paper, fractal clusters composed of particles diffusing with dipolar interaction are observed, and a model taking both magnetic force and diffusion activation energy into account is presented. The computer simulation results generate clusters similar to those observed. And the measured sizes and dimensions of the experimental results are close to that of simulation. Further investigations on the magnetic energy and cluster size reveal that the dipolar interaction and thermal disruption play significant roles in the aggregation of nanosize magnetic particles and the interaction energy is the main driving force of the formation of the ordered structure. PMID- 11735919 TI - Binary hard-sphere crystals with the cesium chloride structure. AB - The possibility of binary hard-sphere colloids crystallizing with the cesium chloride (CsCl) structure was examined experimentally using poly (methyl methacrylate) particles dispersed in organic solvents. Towards this end, two dispersions were prepared that contained particles with a radius ratio alpha=R(B)/R(A), where A refers to the large particles and B the small, of 0.736. This is close to the optimum ratio of 0.732 at which this structure is predicted to form as determined by space-filling calculations. The phases found within the binary mixture were examined using laser light crystallography and scanning electromicroscopy, and some were shown to have the CsCl structure. Over a period of time some of the CsCl crystals disappeared indicating that they were metastable and that this structure may not be the most enduring phase at this size ratio. PMID- 11735920 TI - Experimental test of two-dimensional melting through disclination unbinding. AB - A two-dimensional (2D) melting transition has been studied in a nonequilibrium experimental model system. The system used was a complex or dusty plasma consisting of microspheres suspended in a glow-discharge plasma, where we have mapped the topological defects during the transition. The role of the defects in the melting transition is evaluated and the arrangement of the defects in the lattice is quantified in a new way. It is found that defect density increases dramatically during the melting; at all stages the defects tend to be clustered together rather than widely dispersed; the clustering tends to take the form of chain or string-like structures. We compare these results for the defect structure with the assumptions of the popular 2D melting theory of Halperin and Nelson, rather than the predictions, as is more common. PMID- 11735921 TI - Statistics of largest cluster growth through constant rate random filling of lattices. AB - In this paper we consider a percolation model where the probability p for a site to be occupied increases linearly in time, from 0 to 1. We analyze the way the largest cluster grows in time, and in particular, we study the statistics of the "jumps" in the mass of the largest cluster, and of the time delay between those events. Different critical behaviors are observed below and above the percolation threshold. We propose a theoretical analysis, and we check our results against direct numerical simulations. PMID- 11735922 TI - Spherical polyelectrolyte brushes: comparison between annealed and quenched brushes. AB - We report on a study of spherical polyelectrolyte brushes that consist of a solid core onto which linear polyelectrolyte chains are chemically grafted. The core particles are made up of solid poly(styrene) and have a radius R of ca. 50 nm. As polyelectrolyte chains the weak polyelectrolyte poly(acrylic acid) or the strong polyelectrolyte poly(styrenesulfonate) was used. These chains were generated directly on the surface of the core particles by a grafting-from technique. Hence, the chains are chemically bound to the surface but can be cleaved off and analyzed separately. The contour length L(c) and the number of grafted chains per unit area sigma can thus be determined accurately. The thickness L of the brush layer on the surface has been determined by dynamic light scattering. It is measured for different L(c)/R as a function of pH, ionic strength, and valency of counterions. Annealed brushes exhibit a transition with increasing pH in which the chains are stretched to nearly full length. This can be traced back to the building up of the osmotic pressure of the counterions. The brush height L decreases considerably with increasing ionic strength, most notably when adding divalent ions. The entire set of L as a function of R, L(c), and sigma can be fully explained in terms of a simple two-parameter theory developed by Hariharan et al. [Macromolecules 31, 7514 (1998)]. PMID- 11735923 TI - Adsorption of a diatomic molecular fluid into random porous media. AB - Structural and thermodynamic properties of a homonuclear hard dumbbell fluid adsorbed into a disordered hard sphere matrix are studied by means of integral equation techniques and computer simulation. In particular, we have rewritten the replica Ornstein-Zernike equations to deal with orientational degrees of freedom and we have solved them in two different approaches: the hypernetted chain equation and a semiempirical extension of Verlet's approximation. We have also derived direct expressions to calculate the chemical potential in these approximations. Comparison with grand canonical Monte Carlo results shows that both theoretical treatments describe adequately the physical behavior of the system, Verlet's approach being, however, clearly superior in accordance with previous findings for equilibrated hard core mixtures. PMID- 11735924 TI - Thermal force induced by the presence of a particle near a solidifying interface. AB - The presence of a foreign particle in the melt, ahead of a solid-liquid interface, leads to the onset of interfacial deformations if the thermal conductivity of the particle, k(p), differs from that of the melt, k(l). In this paper, the influence of the thermal conductivity contrast on the interaction between the solidifying interface and the particle is quantified. We show that the interface distortion gives rise to a thermal force whose expression is given by F(th)=2piLGa3(1-alpha)/(2+alpha)T(m), where L is the latent heat of fusion per unit volume, T(m) is the melting point, a is the particle's radius, G the thermal gradient in the liquid phase and alpha=k(p)/k(l). The derivation makes use of the following assumptions: (i) the particle is small compared to the horizontal extent of the interface, (ii) the particle is placed in the near proximity of the deformable solid-liquid interface, and (iii) the interface is practically immobile in the calculation of the thermal field, i.e., V< glass transition for the GdCl3 3AlCl3 glass-forming mixture. AB - The dynamics of density and orientation fluctuations for a member of a family of inorganic glass-forming molten salts based on rare-earth halide-aluminum halide mixtures, namely, the GdCl3-3AlCl3, has been investigated in the supercooled state by means of dynamic light scattering. The temperature dependence of the structural relaxation time near the glass transition temperature was found to be rather strong, indicating the "fragile" character of this liquid. The microscopic origin of the structural relaxation has been rationalized by employing structural information that has been independently extracted from other spectroscopic techniques. PMID- 11735927 TI - Microscopic dynamics of molecular liquids and glasses: role of orientations and translation-rotation coupling. AB - We investigate the dynamics of a fluid of dipolar hard spheres in its liquid and glassy phases, with emphasis on the microscopic time or frequency regime. This system shows rather different glass transition scenarios related to its rich equilibrium behavior, which ranges from a simple hard sphere fluid to long range ferroelectric orientational order. In the liquid phase close to the ideal glass transition line and in the glassy regime a medium range orientational order occurs leading to a softening of an orientational mode. To investigate the role of this mode we use the molecular mode-coupling equations to calculate the spectra straight phi"lm(q,omega) and chi"lm(q,omega). In the center of mass spectra straight phi"00(q,omega) and chi"00(q,omega) we found, besides a high frequency peak at omega(hf), a peak at omega(op), about one decade below omega(hf) x omega(op) has almost no q dependence and exhibits an "isotope" effect omega(op) proportional to I(-1/2), with I the moment of inertia. We give evidence that the existence of this peak is related to the occurrence of medium range orientational order. It is shown that some of these features also exist for schematic mode coupling models. PMID- 11735928 TI - Computer simulations of electrorheological fluids in the dipole-induced dipole model. AB - We have employed the multiple image method to compute the interparticle force for a polydisperse electrorheological (ER) fluid in which the suspended particles can have various sizes and different permittivities. The point-dipole (PD) approximation, being routinely adopted in the computer simulation of ER fluids, is known to err considerably when the particles approach and finally touch due to multipolar interactions. The PD approximation becomes even worse when the dielectric contrast between the particles and the host medium is large. From the results, we show that the dipole-induced-dipole (DID) model yields very good agreements with the multiple image results for a wide range of dielectric contrasts and polydispersity. As an illustration, we have employed the DID model to simulate the athermal aggregation of particles in ER fluids, both in uniaxial and rotating fields. We find that the aggregation time is significantly reduced. The DID model partially accounts for the multipolar interaction and is simple to use in the computer simulation of ER fluids. PMID- 11735929 TI - Solid-solid transformations in a confined soft sphere fluid. AB - Grand canonical Monte Carlo simulations are used to investigate the structure of frozen phases in a confined soft sphere (Lennard-Jones) fluid for a bulk state point lying close to the liquid-solid freezing line. The study reveals that the pore fluid not only freezes in the pore, but solid-solid transformations occur in the two, and three-layered structures that form as the pore height is varied. Using the in-plane bond angle order parameters and pair-correlation functions, the frozen structures and transformations are related to the variations in solvation force and pore density. Transformations from square to triangular lattices are observed in both the two- and three-layered regimes. In the three layered regime, transformations occur from the body-centered tetragonal to hcp and fcc lattices as the pore height is increased. During the transition from one to two layers, random buckled structures are observed. The structural transformations from square to triangular lattices in the confined fluid also result in a splitting in the solvation force curve. PMID- 11735930 TI - Energy landscape view of fracture and avalanches in disordered materials. AB - Molecular simulations are carried out to probe how strain-induced changes in the energy landscape are related to fracture processes in disordered systems. The simulations address a two-dimensional system that consists of 9952 particles with a distribution of sizes, and the changes in the structure and properties with strain are determined with the system constrained to an energy minimum. As the system is strained, local minima of the energy landscape are found to flatten out and disappear, which causes discontinuous structural rearrangements. These structural rearrangements, which correspond to avalanche events, lead to void nucleation and crack growth in discrete steps. PMID- 11735931 TI - Interfacial conditions during evaporation or condensation of water. AB - Steady-state evaporation and condensation experiments have been conducted with water under conditions where buoyancy-driven convection is not present. The temperature profile in each phase has been measured. At the interface, independently of the direction of the phase change, a temperature discontinuity has been found to exist in which the interfacial vapor temperature is greater than that in the liquid. In a thin layer immediately below the interface the temperature is uniform in a layer ( approximately 0.5 mm) and below that the temperature profile is linear, indicating thermal conduction. The uniform temperature layer indicates a mixing process occurs near the interface that could result from surface-tension driven (Marangoni-Benard) convection and/or from "energy partitioning" that is necessary to account for the measured temperature discontinuity near the interface. When the measured interfacial properties are used with the expression for the phase change rate that is obtained from statistical rate theory, it is found that the predictions are in close agreement with the measurements. PMID- 11735932 TI - Kinetics and morphologies of viscoelastic phase separation. AB - In this paper, the effects of relaxational bulk modulus and the average composition of polymers on the viscoelastic phase separation are investigated in detail. It is found that there are two typical morphologies, i.e., moving droplet phase and phase inversion, and the relaxation of the dynamical asymmetry and the amplification of the concentration fluctuation are responsible for the appearance and evolution of different morphologies of viscoelastic phase separation. It is found that, for the viscoelastic phase separation, the scattering function has two peaks. The growth exponents of the main and the secondary peaks in the late stage are almost the same and approximately 0.6, which also agrees with the experimental observations. On the other hand, the growth exponent of the secondary peak increased from approximately 0.42 to approximately 0.66 with increase of straight phi(0) from 0.275 to 0.4, in the intermediate stage. PMID- 11735933 TI - Density change effects on crystal growth from the melt. AB - When a crystal grows from its undercooled melt the local density changes, driving a convective flow in the liquid phase. Then, the purely diffusional description of the process ceases to be satisfactory. Moreover, the dynamic pressure associated with the flow field may affect the melting temperature (and the effective undercooling) of the system. Both these effects have been addressed in recent experimental work. In the present study we derive a thermodynamically consistent phase-field model that accounts for the density change effects in the solidification of a pure substance. Starting from a thermodynamic potential that includes squared gradient terms for both the order parameter and the density, the field equations are derived assuming positive local entropy production. The model is numerically solved in one dimension to show deviations from the classic phase field description of the same phenomenon. PMID- 11735934 TI - Gas spreading on a heated wall wetted by liquid. AB - This study deals with a simple pure fluid whose temperature is slightly below its critical temperature and whose density is nearly critical, so that the gas and liquid phases coexist. Under equilibrium conditions, such a liquid completely wets the container wall and the gas phase is always separated from the solid by a wetting film. We report a striking change in the shape of the gas-liquid interface influenced by heating under weightlessness where the gas phase spreads over a hot solid surface showing an apparent contact angle larger than 90 degrees. We show that the two-phase fluid is very sensitive to the differential vapor recoil force and give an explanation that uses this nonequilibrium effect. We also show how these experiments help to understand the boiling crisis, an important technological problem in high-power boiling heat exchange. PMID- 11735935 TI - Structure formation from mesoscopic soft particles. AB - In this work, the aggregation of mesoscopic gel particles (soft colloids) has been experimentally investigated. The interaction between particles was controlled through the addition of salt, above the critical coagulation concentration, resulting in aggregation with finite bond energies. Attention has been paid to the structure of the clusters formed in the process as well as to the aggregation kinetics. The results indicate that the clusters are fractal and the kinetics of aggregation can be described through the dynamic scaling solution of the Smoluchowski equation. As the energy minimum increases in depth the resultant clusters pass from a very compact structure to typical diffusion limited cluster aggregation (DLCA) fractal dimension values. In addition, the kinetics of growth change from those observed in reaction controlled aggregation to DLCA. These results can be explained within the framework of a reversible growth model, arising from the fact that aggregation takes place in an energy minimum of restricted depth. Moreover, they show that structure and kinetics decouple for such a soft sphere system, in contrast to what is encountered for DLCA and reaction-limited processes. Finally, an unexpected return to a reaction controlled aggregation kinetics was observed for sufficiently deep energy minima, which could be due to the polymerlike particularities of the soft particles considered in this work. PMID- 11735936 TI - Roughness scaling in cyclical surface growth. AB - The scaling behavior of cyclical growth (e.g., cycles of alternating deposition and desorption primary processes) is investigated theoretically and probed experimentally. The scaling approach to kinetic roughening is generalized to cyclical processes by substituting the number of cycles n for the time. The roughness is predicted to grow as n(beta) where beta is the cyclical growth exponent. The roughness saturates to a value that scales with the system size L as L(alpha), where alpha is the cyclical roughness exponent. The relations between the cyclical exponents and the corresponding exponents of the primary processes are studied. Exact relations are found for cycles composed of primary linear processes. An approximate renormalization group approach is introduced to analyze nonlinear effects in the primary processes. The analytical results are backed by extensive numerical simulations of different pairs of primary processes, both linear and nonlinear. Experimentally, silver surfaces are grown by a cyclical process composed of electrodeposition followed by 50% electrodissolution. The roughness is found to increase as a power law of n, consistent with the scaling behavior anticipated theoretically. Potential applications of cyclical scaling include accelerated testing of rechargeable batteries and improved chemotherapeutic treatment of cancerous tumors. PMID- 11735937 TI - Interface pinning in spontaneous imbibition. AB - Evaporation and gravity induced pinning in spontaneous imbibition are examined within a phase field formalism. Evaporation is introduced via a nonconserving term and gravity through a convective term that constrains the influx of liquid. Their effects are described by dimensionless coupling constants epsilon and g, respectively. From liquid conservation, the early time behavior of the average interface position follows H(t) approximately t1/2 until a crossover time t*(g,epsilon). After that the pinning height Hp(g,epsilon) is approached exponentially in time, in accordance with mean field theory. The statistical roughness of the interface is described by an exponent chi approximately 1.25 at all stages of the rise, but the dynamic length scale controlling roughness crosses over from xi(x) approximately H1/2 to a time independent pinning length scale xi(p)(epsilon,g). PMID- 11735938 TI - Van der Waals interaction mediated by an optically uniaxial layer. AB - We study the van der Waals interaction between macroscopic bodies separated by a thin anisotropic film with a uniaxial permittivity tensor. We describe the effect of anisotropy of the media on the magnitude and sign of the interaction. The resulting differences in the van der Waals interaction are especially important for the stability of strongly confined liquid crystals, and nanostructures characterized by highly uniaxial macroscopic molecular arrangement, such as in self-assemblies of long organic molecules forming films, membranes, colloids, etc. We introduce an improved expression for the Hamaker constant which takes into account the uniaxial symmetry of a medium. In special cases neglecting the optical anisotropy even leads to an incorrect sign of the interaction. PMID- 11735939 TI - Pretransitional behavior of dielectric permittivity on approaching a clearing point in a mixture of nematogens with antagonistic configurations of dipoles. AB - The results of studies of static dielectric permittivity epsilon(T) in the isotropic phase of 4-cyano-4-pentylalkylbiphenyl and n-p-methoxybenzylidene-p' butylaniline (5CB-MBBA) mixtures are presented. 5CB and MBBA are nematogens with antagonistic permanent dipole moments configurations. An increase in MBBA concentration strongly decreases the pretransitional effect. However, a derivative analysis detected the existence of a pretransitional anomaly even if its weakness made a straightforward epsilon(T) fit impossible. The obtained anomalies can be well portrayed by fluidlike equations, isomorphic to that applied in critical binary mixtures. Every time the same value of the specific heat critical exponent alpha approximately 0.5 was obtained. A preliminary discussion of the influence of the permanent dipole configuration on the pretransitional behavior of dielectric permittivity was also possible. PMID- 11735940 TI - Computer simulation study of the phase behavior of a nematogenic lattice-gas model. AB - The phase behavior of a mesogenic lattice-gas model consisting of freely rotating spins located at the sites of a three-dimensional cubic lattice has been studied using grand canonical Monte Carlo simulations. When two neighboring sites are occupied, the spin vectors interact via the extensively studied anisotropic Lebwohl-Lasher potential, plus an isotropic term of variable strength. The interaction between occupied and empty sites and two empty sites is taken to be zero. If the parameter governing the strength of the isotropic term is zero, the model exhibits an isotropic fluid-nematic transition, which becomes increasingly stronger as the temperature is lowered. The additional isotropic term is found to be important if the model is to reproduce experimental phase behavior, that is, to exhibit both nematic-vapor coexistence at low temperature and isotropic-vapor coexistence at higher temperatures. PMID- 11735941 TI - Microscopic structure and dynamics of a partial bilayer smectic liquid crystal. AB - Cyanobiphenyls (nCB's) represent a useful and intensively studied class of mesogens. Many of the peculiar properties of nCB's (e.g., the occurrence of the partial bilayer smectic-A(d) phase) are thought to be a manifestation of short range antiparallel association of neighboring molecules, resulting from strong dipole-dipole interactions between cyano groups. To test and extend existing models of microscopic ordering in nCB's, we carry out large-scale atomistic simulation studies of the microscopic structure and dynamics of the Sm-A(d) phase of 4-octyl-4'-cyanobiphenyl (8CB). We compute a variety of thermodynamic, structural, and dynamical properties for this material, and make a detailed comparison of our results with experimental measurements in order to validate our molecular model. Semiquantitative agreement with experiment is found: the smectic layer spacing and mass density are well reproduced, translational diffusion constants are similar to experiment, but the orientational ordering of alkyl chains is overestimated. This simulation provides a detailed picture of molecular conformation, smectic layer structure, and intermolecular correlations in Sm-A(d) 8CB, and demonstrates that pronounced short-range antiparallel association of molecules arising from dipole-dipole interactions plays a dominant role in determining the molecular-scale structure of 8CB. PMID- 11735942 TI - Orientational transitions in a nematic liquid crystal confined by competing surfaces. AB - The effect of confinement on the orientational structure of a nematic liquid crystal model has been investigated by using a version of density-functional theory. We have focused on the case of a nematic confined by opposing flat surfaces, in slab geometry (slit pore), which favor planar molecular alignment (parallel to the surface) and homeotropic alignment (perpendicular to the surface), respectively. The spatial dependence of the tilt angle of the director with respect to the surface normal has been studied, as well as the tensorial order parameter describing the molecular order around the director. For a pore of given width, we find that, for weak surface fields, the alignment of the nematic director is perpendicular to the surface in a region next to the surface favoring homeotropic alignment, and parallel along the rest of the pore, with a sharp interface separating these regions (S phase). For strong surface fields, the director is distorted uniformly, the tilt angle exhibiting a linear dependence on the distance normal to the surface (L phase). Our calculations reveal the existence of a first-order transition between the two director configurations, which is driven by changes in the surface field strength, and also by changes in the pore width. In the latter case the transition occurs, for a given surface field, between the S phase for narrow pores and the L phase for wider pores. A link between the L-S transition and the anchoring transition observed for the semi-infinite case is proposed. PMID- 11735943 TI - Molecular theory of elastic constants of liquid crystals. III. Application to smectic phases with tilted orientational order. AB - Using the density-functional formalism we derive an expression for the distortion free energy of systems with continuous broken symmetry and use it to derive an expression for the elastic constants of smectic phases in which the director is tilted with respect to the smectic layer normal. As in the previous papers of this series [Y. Singh, S. Singh, and K. Rajesh, Phys. Rev. A 45, 974 (1992); Y. Singh, K. Rajesh, V. J. Menon, and S. Singh, Phys. Rev. E 49, 501 (1994)], the expressions for the elastic constants are written in terms of order and structural parameters. The structural parameters involve the generalized spherical harmonic coefficients of the direct pair correlation function of an effective isotropic liquid. The density of this effective isotropic liquid depends on the nature and amount of ordering present in the system and is evaluated self-consistently. We estimate the value of elastic constants using reasonable guesses for the order and structural parameters. PMID- 11735944 TI - Optical rotation and structure of ferrielectric smectic phases. AB - We analyze the measured optical rotation in three- and four-layer smectic ferrielectric phases within the matrix approach to the light propagation. We show that "perfect" three- and four-layer structures with 120 degrees and 90 degrees phase rotation of the director in neighboring layers give negligible optical rotation of polarized light travelling along the normal to the smectic layers. Significant optical rotation is obtained in deformed three- and four-layer smectic phases. The analysis of the measured optical rotatory power clearly shows that three-layer ferrielectric phases of [4-(1-methylheptyloxycarbonylphenyl) octylbiphenyl-4-carboxylate] and 4-[(4-[[1(*) methyl]heptycarboxy]phenyl)carboxy]phenyl-4'-decyloxy-1-benzencarbatioate (10OTBBB1M7) are deformed with the deformation angle of 35 degrees-45 degrees. The deformation angle in the four-layer smectic phase of 10OTBBB1M7 is 70 degrees 90 degrees. This is in reasonable agreement with other experiments and suggests the validity of the "deformed clock model." PMID- 11735945 TI - Experimental evidence for an achiral orthogonal biaxial smectic phase without in plane order exhibiting antiferroelectric switching behavior. AB - We report unambiguous experimental evidence for an achiral orthogonal biaxial smectic-A phase which exhibits antiferroelectric switching behavior. The evidence is based on x-ray-diffraction measurements, texture observation, and the results of dielectric and electro-optical measurements. PMID- 11735946 TI - Dielectric investigation of electrically oriented ferroelectric smectic mixture CS-1013. AB - From dielectric spectroscopic study, a first-order ferroelectric phase transition has been observed in ferroelectric smectic mixture CS-1013 having the phase sequence Cr-SmC*-SmA-N*-Iso. Frequency (100 Hz-10 MHz) and temperature-dependent dielectric measurements have been performed on an electrically aligned sample (thickness 15+/-1 microm) gold coated on glass plates. In the unidirectionally aligned sample, two dielectric relaxation modes (Goldstone mode and soft mode) have been clearly observed in the ferroelectric SmC* phase while only one relaxation mode (soft mode) is visualized in the paraelectric SmA phase. Low frequency molecular relaxation was also observed in the smectic phases. The experimental results have also been analyzed at different temperatures and biasing voltages for an understanding of the dynamics of dielectric processes in the ferroelectric phase. Finally, we proposed the "pseudospin" model for understanding the ferroelectric-antiferroelectric transition in liquid crystals. We associate the tilt angle straight theta and the pitch of the helix, respectively, with biaxial (b) and uniaxial (u) anisotropy parameters as fluctuating parameters around their stability limit (corresponding to the crystalline values). Here, the director acts as the pseudospin variable. This gives rise to a transverse Ising type (or anisotropic Heisenberg model under the mean-field approximation). It is then shown that such a model with fluctuations of (b) and (u) would explain the ferroelectric and antiferroelectric phase transitions in such liquid crystals. Using Landau theory and the stability conditions, we have also shown, in brief, the feasibility of different types of phase transitions in the ferroelectric liquid crystal system. PMID- 11735947 TI - Photo-orientation of liquid crystals due to light-induced desorption and adsorption of dye molecules on an aligning surface. AB - We show that adsorption of dye molecules control the light-induced alignment of dye-doped nematic liquid crystal (LC) on a nonphotosensitive polymer surface. The dependencies of light-induced twist structures on exposure, thermal baking, thickness, and aging before irradiation of the LC cells allowed us to propose the following mechanism for the alignment. Before irradiation, the "dark"-adsorbed layer on the tested surface is formed from dye molecules predominantly aligned along the initial direction of the director. Irradiation of the cell with linearly polarized light produces an additional layer with different orientational ordering of dye molecules. The final easy axis is determined by the competition of "dark" and light-induced contributions to anchoring and is aligned between the "dark" easy axes and polarization of the light. For quantitative interpretation, we apply the tensor model of anchoring and assume that the photoalignment in the mesophase is a cumulative effect of the light-induced anchoring on the background of the already existing anisotropic "dark" dye layer. PMID- 11735948 TI - Nematic model in the presence of a finite disorienting field: integral equation approach. AB - A fluid of uniaxial particles in a disorienting field is considered as a simple model of biaxial nematics. The model stability with respect to the spontaneous formation of a biaxial phase is investigated by means of the integral equation method. The orientational instability condition is obtained explicitly and turns into known results for the limiting cases of zero and of infinite fields. It is shown that the biaxiality induced by small fields can expand considerably the region of spontaneously ordered fluid and could be useful to obtain mesomorphic phases in nonmesogens. The effect of small disorienting fields is more pronounced in systems with short-range anisotropic interactions between particles. PMID- 11735949 TI - Forces in the isotropic phase of a confined nematic liquid crystal 5CB. AB - Using a temperature controlled atomic force microscope, we have measured the temperature dependence of the force between a flat silanated glass surface and a silanated glass microsphere, immersed in the isotropic phase of the nematic liquid crystal 5CB (4'-n-pentyl 4-cyanobiphenyl). At separations of several nanometers, we observed a weak, short range attractive force of the order of 100 pN, which was increased by decreasing the temperature. The temperature dependence of the amplitude and the range of this attractive force can be described by a combination of van der Waals and a mean-field prenematic force due to the surface induced nematic order. This is supported by ellipsometric study and allows for the determination of the surface coupling energy of 5CB on a silanated glass surface. PMID- 11735950 TI - Evidence from infrared dichroism, x-ray diffraction, and atomistic computer simulation for a "zigzag" molecular shape in tilted smectic liquid crystal phases. AB - Infrared dichroism (IR) and atomistic computer simulation are employed to probe molecular shape in smectic liquid crystal phases where the optic axis is tilted relative to the layer normal. Polar plots of absorption profiles due to core (phenyl, C-C) and tail (alkyl or methylene, CH2) vibrations in the tilted synclinic (smectic-C) phase of a variety of materials show the phenyl (core) IR absorbance symmetry axes to be consistently tilted at larger angle from the layer normal than the alkyl or methylene (tail). This suggests that, on average, the tails are less tilted than the cores. Furthermore, we find that optic axis tilt angle is close to the core tilt angle measured by IR dichroism, as expected, since liquid crystal birefringence arises primarily from the cores. These results are in accord with the "zigzag" model of Bartolino, Doucet, and Durand. However, we find that only a small fraction of the tail, the part nearest the core, is tilted, and only this part contributes significantly to layer contraction upon molecular tilt. PMID- 11735951 TI - Detailed optical studies of several banana-shaped compounds in liquid crystal B2 phase. AB - Null-transmission ellipsometry has been performed on free-standing films of one compound in the liquid crystal B2 phase. We have studied films of thickness from 1 to 121 layers. One of the compounds used has an unusually wide 59 K window for the B2 phase. The tilt angle was investigated as a function of temperature and found to be constant over this temperature range to within our resolution of 1 degrees. The one-layer films studied exhibit the same structure as the thicker films, and have helped us to refine the optical model for the B2 phase. For thin films we find that modeling a single smectic layer as two uniaxial layers is a better description of the data than a single biaxial layer, but that for thick films the model used does not effect the simulated result appreciably. Preliminary results also find that the surface layers are less tilted than the interior layers, in contrast to rodlike liquid crystals, which show an enhanced surface tilt. PMID- 11735952 TI - Yield conditions for deformation of amorphous polymer glasses. AB - Shear yielding of glassy polymers is usually described in terms of the pressure dependent Tresca or von Mises yield criteria. We test these criteria against molecular dynamics simulations of deformation in amorphous polymer glasses under triaxial loading conditions that are difficult to realize in experiments. Difficulties and ambiguities in extending several standard definitions of the yield point to triaxial loads are described. Two definitions, the maximum and offset octahedral stresses, are then used to evaluate the yield stress for a wide range of model parameters. In all cases, the onset of shear is consistent with the pressure-modified von Mises criterion, and the pressure coefficient is nearly independent of many parameters. Under triaxial tensile loading, the mode of failure changes to cavitation, and the von Mises criterion no longer applies. PMID- 11735953 TI - Probing the subglass relaxation behavior in model heterocyclic polymer networks by dielectric spectroscopy. AB - The subglass relaxation (beta) in model heterocyclic polymer networks (HPNs) with a controlled ratio of trimerized mono- and diisocyanates was characterized by dielectric spectroscopy in the frequency domain. The beta relaxation in the investigated HPNs follows the Arrhenius law with unusually low values of the preexponential factor (10(-17)C reaction-diffusion system with initially-separated reactants. The experimental setup consists of a CCD camera monitoring the kinetics of the front formed in the reaction-diffusion process Cu(2+) + tetra [disodium ethyl bis(5-tetrazolylazo) acetate trihydrate] -->1:1 complex, in aqueous, gel-free solution, taking place inside a 150 microm gap between two flat microscope slides. The experimental results agree with the theoretical predictions for the time dependence of the front's width, height, and location, as well as the global reaction rate. PMID- 11735999 TI - Mandelbrot set in coupled logistic maps and in an electronic experiment. AB - We suggest an approach to constructing physical systems with dynamical characteristics of the complex analytic iterative maps. The idea follows from a simple notion that the complex quadratic map by a variable change may be transformed into a set of two identical real one-dimensional quadratic maps with a particular coupling. Hence, dynamical behavior of similar nature may occur in coupled dissipative nonlinear systems, which relate to the Feigenbaum universality class. To substantiate the feasibility of this concept, we consider an electronic system, which exhibits dynamical phenomena intrinsic to complex analytic maps. Experimental results are presented, providing the Mandelbrot set in the parameter plane of this physical system. PMID- 11736000 TI - Spatiotemporal patterns in CO oxidation on Pt(110): the role of nonlinear diffusion. AB - Standing-wave patterns observed in the CO + O2 reaction on Pt(110) are described by a model that explicitly takes into account the coupling between the transport of adsorbed CO and the adsorbate-induced structural transformation of the substrate. We show that synchronization of the surface is achieved through nucleation and growth processes even in the absence of gas-phase coupling. PMID- 11736001 TI - Golden rule decay versus Lyapunov decay of the quantum Loschmidt echo. AB - The overlap of two wave packets evolving in time with slightly different Hamiltonians decays exponentially approximate to e(-gammat), for perturbation strengths U greater than the level spacing Delta. We present numerical evidence for a dynamical system that the decay rate gamma is given by the smallest of the Lyapunov exponent lambda of the classical chaotic dynamics and the level broadening U(2)/Delta that follows from the golden rule of quantum mechanics. This implies the range of validity U > the square root of [lambdaDelta] for the perturbation-strength independent decay rate discovered by Jalabert and Pastawski [Phys. Rev. Lett. 86, 2490 (2001)]. PMID- 11736002 TI - Synchronization using dynamic coupling. AB - A systematic coupling procedure is introduced for synchronizing arbitrary chaotic dynamical systems. This coupling exploits the existing contraction properties of the flow and suppresses divergence only along those directions in state space, where the underlying flow is not contracting. In this way, systems can be synchronized using a minimum of transmitted information for guaranteed high quality synchronization. Applications in combination with sporadic driving and in partitioned state spaces are numerically illustrated. PMID- 11736003 TI - Relaxation, the Boltzmann-Jeans conjecture, and chaos. AB - Slow (logarithmic) relaxation from a highly excited state is studied in a Hamiltonian system with many degrees of freedom. The relaxation time is shown to increase as the exponential of the square root of the energy of excitation, in agreement with the Boltzmann-Jeans conjecture, while it is found to be inversely proportional to residual Kolmogorov-Sinai entropy, introduced in this Rapid Communication. The increase of the thermodynamic entropy through this relaxation process is found to be proportional to this quantity. PMID- 11736004 TI - Comparison of averages of flows and maps. AB - It is shown that in transient chaos there is no direct relation between averages in a continuous time dynamical system (flow) and averages using the analogous discrete system defined by the corresponding Poincare map. In contrast to permanent chaos, results obtained from the Poincare map can even be qualitatively incorrect. The reason is that the return time between intersections on the Poincare surface becomes relevant. However, after introducing a true-time Poincare map, quantities known from the usual Poincare map, such as conditionally invariant measure and natural measure, can be generalized to this case. Escape rates and averages, e.g., Liapunov exponents and drifts, can be determined correctly using these measures. Significant differences become evident when we compare with results obtained from the usual Poincare map. PMID- 11736005 TI - Energy of internal modes of nonlinear waves and complex frequencies due to symmetry breaking. AB - Considering a class of Hamiltonian systems, it is demonstrated that energy of the internal modes with real frequencies supported by nonlinear waves and appearing due to perturbations breaking a continuous symmetry has its sign determined by the symmetry itself, independently of the nature of the perturbations. In particular, it is shown that negative energy modes emerge as a result of the breaking of the phase symmetry in the perturbed nonlinear-Schrodinger equation. An expression for energy of the Vakhitov-Kolokolov internal modes is also derived. Comparative analysis of the energy signs of the internal modes in these two cases explains the ubiquity of instabilities with complex frequencies of solitary and continuous waves in systems with broken phase symmetry. PMID- 11736006 TI - Superluminal optical pulse propagation at 1.5 microm in periodic fiber Bragg gratings. AB - We report on the experimental observation of superluminal tunneling of picosecond optical pulses in a periodic fiber Bragg grating. Optical pulses of 380-ps duration, generated by an externally-modulated single-frequency erbium-ytterbium laser operating near 1.5-microm wavelength, were propagated at a group velocity greater than approximately 1.97 times the speed of light in vacuum across a 2-cm long fiber grating. Owing to the very large ratio between the thickness of the barrier (2 cm) and the wavelength of probing optical pulses ( approximately 1.5 microm), our experiment allows for the observation of superluminal tunneling in the optical region by direct optoelectronic time-domain measurements. PMID- 11736007 TI - Ministop bands in single-defect photonic crystal waveguides. AB - We numerically study single-defect photonic crystal waveguides obtained from a triangular lattice of air holes in a dielectric background. It is found that, for medium-high air filling ratios, the transmission has very small values in narrow frequency regions lying inside the photonic band gap-the so-called ministop bands. Two types of ministop bands are shown to exist; one of which is due to the multimode nature of the waveguide. Their dependence on the length of the waveguide and on the air filling ratio is presented. PMID- 11736008 TI - Determining the density of states for classical statistical models: a random walk algorithm to produce a flat histogram. AB - We describe an efficient Monte Carlo algorithm using a random walk in energy space to obtain a very accurate estimate of the density of states for classical statistical models. The density of states is modified at each step when the energy level is visited to produce a flat histogram. By carefully controlling the modification factor, we allow the density of states to converge to the true value very quickly, even for large systems. From the density of states at the end of the random walk, we can estimate thermodynamic quantities such as internal energy and specific heat capacity by calculating canonical averages at any temperature. Using this method, we not only can avoid repeating simulations at multiple temperatures, but we can also estimate the free energy and entropy, quantities that are not directly accessible by conventional Monte Carlo simulations. This algorithm is especially useful for complex systems with a rough landscape since all possible energy levels are visited with the same probability. As with the multicanonical Monte Carlo technique, our method overcomes the tunneling barrier between coexisting phases at first-order phase transitions. In this paper, we apply our algorithm to both first- and second-order phase transitions to demonstrate its efficiency and accuracy. We obtained direct simulational estimates for the density of states for two-dimensional ten-state Potts models on lattices up to 200 x 200 and Ising models on lattices up to 256 x 256. Our simulational results are compared to both exact solutions and existing numerical data obtained using other methods. Applying this approach to a three-dimensional +/-J spin-glass model, we estimate the internal energy and entropy at zero temperature; and, using a two-dimensional random walk in energy and order parameter space, we obtain the (rough) canonical distribution and energy landscape in order-parameter space. Preliminary data suggest that the glass transition temperature is about 1.2 and that better estimates can be obtained with more extensive application of the method. This simulational method is not restricted to energy space and can be used to calculate the density of states for any parameter by a random walk in the corresponding space. PMID- 11736009 TI - Monomer-dimer reaction model with asymmetric adsorption of monomer and dimer on the catalyst surface. AB - The irreversible kinetics of the Ziff-Gulari-Barshad model in the presence of inhomogeneity on a catalyst surface is investigated by means of Monte Carlo simulation. We assume that only part of catalyst surface sites are active for surface catalytic reaction and randomly distributed on the catalyst surface. The adsorption of a O2 molecule is permitted if one site of a nearest-neighbor vacant pair is active. CO can adsorb on all vacant sites, but the adsorption probability is p(p<1) if the site is not active. In our modified model, the O-passivated phase disappears and the continuous phase transition between the O passivated and the reactive state is eliminated. We also find that the transition between the CO passivated and the reactive state is continuous if the concentration of the active sites is small, and it becomes discontinuous with increasing concentrations of active sites. Furthermore, it is shown that a hysteresis loop exists whether the CO-passivated transition is continuous or discontinuous. Our simulation results are in good agreement with many relevant experimental results and may provide an alternative explanation for the experimental observations. PMID- 11736010 TI - Phase transitions in systems with 1/r(alpha) attractive interactions. AB - Collapse, or a gravitational-like phase transition is studied in a microcanonical ensemble of particles with an attractive 1/r(alpha) potential. A mean-field continuous integral equation is used to determine a saddle-point density profile that extremizes the entropy functional. For all 0 < alpha < 3, a critical energy is determined below which the entropy of the system exhibits a discontinuous jump. If an effective short-range cutoff is applied, the entropy jump is finite; if not, the entropy diverges to +infinity. A stable integral equation solution represents a state with maximal entropy; the reverse is always true only for a modified integral equation introduced here. PMID- 11736011 TI - Critical behavior of a one-dimensional fixed-energy stochastic sandpile. AB - We study a one-dimensional fixed-energy version (that is, with no input or loss of particles) of Manna's stochastic sandpile model. The system has a continuous transition to an absorbing state at a critical value of the particle density, and exhibits the hallmarks of an absorbing-state phase transition, including finite size scaling. Critical exponents are obtained from extensive simulations, which treat stationary and transient properties, and an associated interface representation. These exponents characterize the universality class of an absorbing-state phase transition with a static conserved density in one dimension; they differ from those expected at a linear-interface depinning transition in a medium with point disorder, and from those of directed percolation. PMID- 11736012 TI - Conductivity of continuum percolating systems. AB - We study the conductivity of a class of disordered continuum systems represented by the Swiss-cheese model, where the conducting medium is the space between randomly placed spherical holes, near the percolation threshold. This model can be mapped onto a bond percolation model where the conductance sigma of randomly occupied bonds is drawn from a probability distribution of the form sigma(-a). Employing the methods of renormalized field theory we show to arbitrary order in epsilon expansion that the critical conductivity exponent of the Swiss-cheese model is given by t(SC)(a) = (d-2)nu + max[phi,(1-a)(-1)], where d is the spatial dimension and nu and phi denote the critical exponents for the percolation correlation length and resistance, respectively. Our result confirms a conjecture that is based on the "nodes, links, and blobs" picture of percolation clusters. PMID- 11736013 TI - Multibaker map for shear flow and viscous heating. AB - A consistent description of shear flow and the accompanying viscous heating as well as the associated entropy balance is given in the framework of a deterministic dynamical system. The laminar shear flow is modeled by a Hamiltonian multibaker map which drives velocity and temperature fields. In the appropriate macroscopic limit one recovers the Navier-Stokes and heat conduction equations along with the associated entropy balance. This indicates that results of nonequilibrium thermodynamics can be described by means of an abstract, sufficiently chaotic, and mixing dynamics. A thermostating algorithm can also be incorporated into this framework. PMID- 11736014 TI - Number of times a site is visited in two-dimensional random walks. AB - In this paper, formulas are derived to compute the mean number of times a site has been visited in a random walk on a two-dimensional lattice. Asymmetric random walks are considered, with or without drift, for different boundary conditions. It is shown that in case of absorbing boundaries the mean number of visits reaches stationary values over the lattice; comparisons with a Monte Carlo simulation are also presented. PMID- 11736015 TI - Elastic properties of Sierpinski-like carpets: finite-element-based simulation. AB - The elastic properties of two-dimensional continuous composites of fractal structures are studied with the set of Sierpinski-like carpets filled by voids or rigid inclusions. The effective elastic moduli of these carpets are calculated numerically using the finite-element and position-space renormalization group techniques. The fixed-point problem is analyzed by flow diagrams in the plane of the current Poisson ratios and coefficients of anisotropy of the composites. It is found that in the general case the effective elastic moduli asymptotically approach a power-law behavior. Moreover, the common exponent characterizes the scaling behavior of each component of the elastic modulus tensor of a definite carpet. The values of the scaling exponents and positions of the fixed points are shown to be independent of the elastic properties of the host and depend significantly on the fractal dimension of the composite. PMID- 11736016 TI - Constructing, characterizing, and simulating Gaussian and higher-order point distributions. AB - The definition and the properties of a Gaussian point distribution, in contrast to the well-known properties of a Gaussian random field are discussed. Constraints for the number density and the two-point correlation function arise. A simple method for the simulation of this so-called Gauss-Poisson point process is given and illustrated with an example. A comparison of the distribution of galaxies in the PSCz catalog with the Gauss-Poisson process underlines the importance of higher-order correlation functions for the description for the galaxy distribution. The construction of the Gauss-Poisson point process is extended to the n-point Poisson cluster process, now incorporating correlation functions up to nth order. Simulation methods and constraints on the correlation functions are discussed for an n-point case and detailed for a three-point case. As another approach, well suited for strongly clustered systems, the generalized halo model is discussed. The influence of substructure inside the halos on the two- and three-point correlation functions is calculated in this model. PMID- 11736017 TI - Instability of Alexander-McTague crystals and its implication for nucleation. AB - We show that the argument of Alexander and McTague, that the bcc crystalline structure is favored in those crystallization processes where the first-order character is not too pronounced, is not correct. We find that any solution that satisfies the Alexander-McTague condition is not stable. We investigate the implication of this result for nucleation near the pseudospinodal in near meanfield systems. PMID- 11736018 TI - Thermal conduction in a chain of colliding harmonic oscillators revisited. AB - Thermal conduction in a chain of colliding harmonic oscillators, sometimes called the ding-dong model, is investigated. We first argue that this system is equivalent to the Dawson plasma sheet model. Next we show the Lyapunov analysis for this system to characterize its dynamical property. Finally, we reconsider the numerical study of thermal conduction for this system using the Green-Kubo relation and the direct simulation of Fourier law. Both show that thermal conduction is normal in that kappa(N,T) approximately equals N(0), at least, at low temperature in a large system. PMID- 11736019 TI - Flat histogram simulation of lattice polymer systems. AB - We demonstrate the use of an algorithm called the Flat Histogram sampling algorithm for the simulation of two-dimensional lattice polymer systems. Thermodynamic properties, such as the average energy or entropy and other physical quantities such as the end-to-end distance or radius of gyration can be easily calculated using this method. The ground-state energy can also be determined. We also explore the accuracy and limitations of this method. PMID- 11736020 TI - Macroscopic equations for pattern formation in mixtures of microtubules and molecular motors. AB - Inspired by patterns observed in mixtures of microtubules and molecular motors, we propose continuum equations for the evolution of motor density and microtubule orientation. The chief ingredients are the transport of motors along tubules, and the alignment of tubules in the process. The macroscopic equations lead to aster and vortex patterns in qualitative agreement with experiments. While the early stages of evolution of tubules are similar to coarsening of spins following a quench, the rearrangement of motors leads to arrested coarsening at low densities. Even in one dimension, the equations exhibit a variety of interesting behaviors, such as symmetry breaking, moving fronts, and motor localization. PMID- 11736021 TI - Cluster size distributions in particle systems with asymmetric dynamics. AB - We present exact and asymptotic results for clusters in the one-dimensional totally asymmetric exclusion process (TASEP) with two different dynamics. The expected length of the largest cluster is shown to diverge logarithmically with an increasing system size for ordinary TASEP dynamics and as a logarithm divided by a double logarithm for generalized dynamics, where the hopping probability of a particle depends on the size of the cluster it belongs to. The connection with the asymptotic theory of extreme order statistics is discussed in detail. We also consider a related model of interface growth, where the deposited particles are allowed to relax to the local gravitational minimum. PMID- 11736022 TI - Statistical and dynamical study of disease propagation in a small world network. AB - Statistical properties and dynamical disease propagation have been studied numerically using a percolation model in a one dimensional small world network. The parameters chosen correspond to a realistic network of school age children. It has been found that percolation threshold decreases as a power law as the shortcut fluctuations increase. It has also been found that the number of infected sites grows exponentially with time and its rate depends logarithmically on the density of susceptibles. This behavior provides an interesting way to estimate the serology for a given population from the measurement of the disease growing rate during an epidemic phase. The case in which the infection probability of nearest neighbors is different from that of short cuts has also been examined. A double diffusion behavior with a slower diffusion between the characteristic times has been found. PMID- 11736023 TI - Exactly solvable models through the empty-interval method. AB - The most general one dimensional reaction-diffusion model with nearest-neighbor interactions that can be solved exactly through empty-interval method has been introduced. Assuming translationally invariant initial conditions, the probability that n consecutive sites are empty, E(n), has been exactly obtained. Here, however, we do not consider reactions changing two empty neighboring sites. In the thermodynamic limit, the large-time behavior of the system has also been investigated. Releasing translationally invariance, the evolution equation for the probability that n consecutive sites, starting from the site k, are empty, E(k,n), is obtained. In the thermodynamic limit, the large time behavior of the system is also considered. Finally, the continuum limit of the model is considered and the empty-interval probability function is obtained. PMID- 11736024 TI - Breakdown of the Landauer bound for information erasure in the quantum regime. AB - A known aspect of the Clausius inequality is that an equilibrium system subjected to a squeezing dS < 0 of its entropy must release at least an amount the absolute value of dQ = T times the absolute value of dS of heat. This serves as a basis for the Landauer principle, which puts a lower bound T ln 2 for the heat generated by erasure of one bit of information. Here we show that in the world of quantum entanglement this law is broken. A quantum Brownian particle interacting with its thermal bath can either generate less heat or even absorb heat during an analogous squeezing process, due to entanglement with the bath. The effect exists even for weak but fixed coupling with the bath, provided that temperature is low enough. This invalidates the Landauer bound in the quantum regime, and suggests that quantum carriers of information can be more efficient than assumed so far. PMID- 11736025 TI - Percolation and jamming in random bond deposition. AB - A model is presented in which on the bonds of a square lattice linear segments ("needles") of a constant length a are randomly placed. We investigate the dependence of the percolation and jamming thresholds on the length of the needles. The difference from the standard site deposition problem is demonstrated. We show that the system undergoes a transition at a=6. When shorter needles are used, the system first becomes percolating before becoming jammed. For longer needles the lattice becomes jammed but there is no percolation. We present evidence that the transition is due to different clustering of the short and long needles. We also determine the Fisher exponent, obtaining the same value as for standard two-dimensional percolation. PMID- 11736026 TI - Atomic motion in magneto-optical double-well potentials: a testing ground for quantum chaos. AB - We have identified ultracold atoms in magneto-optical double-well potentials as a very clean setting in which to study the quantum and classical dynamics of a nonlinear system with multiple degrees of freedom. In this system, entanglement at the quantum level and chaos at the classical level arise from nonseparable couplings between the atomic spin and its center of mass motion. The main features of the chaotic dynamics are analyzed using action-angle variables and Poincare surfaces of section. We show that for the initial state prepared in current experiments [D. J. Haycock et al., Phys. Rev. Lett. 85, 3365 (2000)], classical and quantum expectation values diverge after a finite time, and the observed experimental dynamics is consistent with quantum-mechanical predictions. Furthermore, the motion corresponds to tunneling through a dynamical potential barrier. The coupling between the spin and the motional subsystems, which are very different in nature from one another, leads to interesting questions regarding the transition from regular quantum dynamics to chaotic classical motion. PMID- 11736027 TI - Quantum criticality for few-body systems: path-integral approach. AB - We present the path-integral approach to treat quantum phase transitions and critical phenomena for few-body quantum systems. Allowing the space and time variables to have discrete values, we turn the quantum problem into an effective classical lattice problem. Imposing the constraint that any change in space time must preserve the scaling invariance of Brownian paths, we show that the mapped classical lattice system has a known scaling behavior when the particle is free, which breaks down when the strength of the interaction potential reaches a certain value. In principle, any quantity with known scaling behavior may be used to determine the transition point. We illustrate the method by numerically evaluating the correlation length and the radial mean distance for a system composed of a single particle in the presence of an attractive Poschl-Teller potential in one and three dimensions. The method is general and has potential applicability for large systems. PMID- 11736028 TI - Energies and collapse times of symmetric and symmetry-breaking states of finite systems with a U(1) symmetry. AB - We study quantum systems of volume V, which will exhibit the breaking of a U(1) symmetry in the limit of V-->infinity, when V is large but finite. We estimate the energy difference between the "symmetric ground state" (SGS), which is the lowest-energy state that does not break the symmetry, and a "pure phase vacuum" (PPV), which approaches a symmetry-breaking vacuum as V-->infinity. Under some natural postulates on the energy of the SGS, it is shown that PPVs always have a higher energy than the SGS, and we derive a lower bound of the excess energy. We argue that the lower bound is O(V0), which becomes much larger than the excitation energies of low-lying excited states for a large V. We also discuss the collapse time of PPVs for interacting many bosons. It is shown that the wave function collapses in a microscopic time scale, because PPVs are not energy eigenstates. We show, however, that for PPVs the expectation value of any observable, which is a finite polynomial of boson operators and their derivatives, does not collapse for a macroscopic time scale. In this sense, the collapse time of PPVs is macroscopically long. PMID- 11736029 TI - Random-bond Potts model in the large-q limit. AB - We study the critical behavior of the q-state Potts model with random ferromagnetic couplings. Working with the cluster representation the partition sum of the model in the large-q limit is dominated by a single graph, the fractal properties of which are related to the critical singularities of the random-Potts model. The optimization problem of finding the dominant graph, is studied on the square lattice by simulated annealing and by a combinatorial algorithm. Critical exponents of the magnetization and the correlation length are estimated and conformal predictions are compared with numerical results. PMID- 11736030 TI - Criticality in a dynamic mixed system. AB - We suggest a dynamic generalization of the simplest static hierarchical mixed model introduced by Shnirman and Blanter [Phys. Rev. Lett. 81, 5445 (1998); Phys. Rev. E. 60, 5111 (1998). We show that the stationary solution of the dynamic mixed model (DMM) demonstrates, in general, a linear form of the magnitude frequency relation and may be considered a self-organized critical system. The dynamic mixed model demonstrates three principal kinds of system behavior: stability, catastrophe, and scale invariance. We show that the catastrophic area exists for all parameters of the mixture, and obtain three analytical expressions for boundary conditions of the stability and the scale invariance domains. As in the static model scale invariance appears as a result of a strong heterogeneity of the mixture. We describe how the magnitude-frequency relation reflects parameters of the heterogeneity and healing conditions for different domains of system behavior. Deviation of the DMM from the static mixed model and possible applications to earthquake prediction are discussed. PMID- 11736031 TI - Dynamics of spiral waves under the modulation of noise pulses. AB - This work aims at investigating the dynamics of spiral waves under the modulation of noise pulses. Both rigid rotating and meandering spirals are considered. The numerical simulations show that for meandering spirals there exists a minimal external radius of the tip trajectory at an optimal intensity when keeping the duration constant, or for an optimal duration when keeping the intensity constant. For rigid rotating spirals an interesting phenomenon is that the clockwise-counterclockwise transition of the trajectory occurs when we raise the duration of the noise pulse for a fixed intensity. PMID- 11736032 TI - Non-Hamiltonian equations of motion with a conserved energy. AB - In 1980 Andersen introduced the use of "extended system" as a means of exploring by molecular dynamics simulation the phase space of a physical model according to a desired ensemble distribution different from the standard microcanonical function. Following his original work on constant pressure-constant enthalpy a large number of different equations of motion, not directly derivable from a Hamiltonian, have been proposed in recent years, the most notable of which is the so-called Nose-Hoover formulation for "canonical" molecular dynamics simulation. Using a generalization of the symplectic form of the Hamilton equations of motion we show here that there is a unique general structure that underlies most, if not all the equations of motion for "extended systems." We establish a unifying formalism that allows one to identify and separately control the conserved quantity, usually known as the "total energy" of the system, and the phase-space compressibility. Moreover, we define a standard procedure to construct conservative non-Hamiltonian flows that sample the phase space according to a chosen distribution function [Tuckerman et al., Europhys. Lett. 45, 149 (1999)]. To illustrate the formalism we derive new equations of motion for two example cases. First we modify the equations of motion of the Nose-Hoover thermostat applied to a one-dimensional harmonic oscillator, and we show how to overcome the ergodicity problem and obtain a canonical sampling of phase space without making recourse to additional degrees of freedom. Finally we recast an idea recently put forward by Marchi and Ballone [J. Chem. Phys. 110, 3697 (1999)] and derive a dynamical scheme for sampling phase space with arbitrary statistical biases, showing as an explicit application a demixing transition in a simple Lennard Jones binary mixture. PMID- 11736033 TI - Macroscopic traffic models from microscopic car-following models. AB - We present a method to derive macroscopic fluid-dynamic models from microscopic car-following models via a coarse-graining procedure. The method is first demonstrated for the optimal velocity model. The derived macroscopic model consists of a conservation equation and a momentum equation, and the latter contains a relaxation term, an anticipation term, and a diffusion term. Properties of the resulting macroscopic model are compared with those of the optimal velocity model through numerical simulations, and reasonable agreement is found although there are deviations in the quantitative level. The derivation is also extended to general car-following models. PMID- 11736034 TI - Dynamic constitutive relations for polarization and magnetization. AB - In this paper we develop constitutive relations for materials where the magnetization and polarization may depend on both the electric and magnetic fields. The approach is general, and is based on a previously developed statistical-mechanical theory. We include the quadrupole-moment density as well as the dipole-moment density in the microscopic displacement field. This yields an electric gradient term in the constitutive equations. This leads to origin invariance in the multipole moments from which Maxwell's equations are defined. We present generalizations of Debye and Landau-Lifshitz equations of motion which are valid for nonequilibrium and contain memory. The reversible and relaxation terms in the polarization and magnetization evolution equations include the possibility of magnetoelectric coupling. Using constitutive relationship, we derive evolution equations for the displacement and induction fields from a Hamiltonian approach. PMID- 11736035 TI - Reparametrization-covariant theory for on-line learning of probability distributions. AB - We discuss the on-line learning of probability distributions in a reparametrization covariant formulation. Reparametrization covariance plays an essential role not only to respect an intrinsic property of "information" but also for pattern recognition problems. We can obtain an optimal on-line learning algorithm with reparametrization invariance, where the conformal gauge connects a covariant formulation with a noncovariant one in a natural way. PMID- 11736036 TI - Thermodynamic entropy production fluctuation in a two-dimensional shear flow model. AB - We investigate fluctuations in the momentum flux across a surface perpendicular to the velocity gradient in a stationary shear flow maintained by either thermostated deterministic or by stochastic boundary conditions. In the deterministic system the fluctuation relation for the probability of large deviations, which holds for the phase space volume contraction giving the Gibbs ensemble entropy production, never seems to hold for the flux which gives the hydrodynamic entropy production. In the stochastic case the fluctuation relation is found to hold for the total flux, as predicted by various exact results, but not for the flux across part of the surface. The latter appear to satisfy a modified fluctuation relation. Similar results are obtained for the heat flux in a steady state produced by stochastic boundaries at different temperatures. PMID- 11736037 TI - Quantum thermodynamic cooling cycle. AB - The quantum-mechanical and thermodynamic properties of a three-level molecular cooling cycle are derived. An inadequacy of earlier models is rectified in accounting for the spontaneous emission and absorption associated with the coupling to the coherent driving field via an environmental reservoir. This additional coupling need not be dissipative, and can provide a thermal driving force-the quantum analog of classical absorption chillers. The dependence of the maximum attainable cooling rate on temperature, at ultralow temperatures, is determined and shown to respect the recently established fundamental bound based on the second and third laws of thermodynamics. PMID- 11736038 TI - Adaptive and self-averaging Thouless-Anderson-Palmer mean-field theory for probabilistic modeling. AB - We develop a generalization of the Thouless-Anderson-Palmer (TAP) mean-field approach of disorder physics, which makes the method applicable to the computation of approximate averages in probabilistic models for real data. In contrast to the conventional TAP approach, where the knowledge of the distribution of couplings between the random variables is required, our method adapts to the concrete set of couplings. We show the significance of the approach in two ways: Our approach reproduces replica symmetric results for a wide class of toy models (assuming a nonglassy phase) with given disorder distributions in the thermodynamic limit. On the other hand, simulations on a real data model demonstrate that the method achieves more accurate predictions as compared to conventional TAP approaches. PMID- 11736039 TI - Optimizing traffic lights in a cellular automaton model for city traffic. AB - We study the impact of global traffic light control strategies in a recently proposed cellular automaton model for vehicular traffic in city networks. The model combines basic ideas of the Biham-Middleton-Levine model for city traffic and the Nagel-Schreckenberg model for highway traffic. The city network has a simple square lattice geometry. All streets and intersections are treated equally, i.e., there are no dominant streets. Starting from a simple synchronized strategy, we show that the capacity of the network strongly depends on the cycle times of the traffic lights. Moreover, we point out that the optimal time periods are determined by the geometric characteristics of the network, i.e., the distance between the intersections. In the case of synchronized traffic lights, the derivation of the optimal cycle times in the network can be reduced to a simpler problem, the flow optimization of a single street with one traffic light operating as a bottleneck. In order to obtain an enhanced throughput in the model, improved global strategies are tested, e.g., green wave and random switching strategies, which lead to surprising results. PMID- 11736040 TI - Origin of scaling structure and non-Gaussian velocity distribution in a self gravitating ring model. AB - Fractal structures and non-Gaussian velocity distributions are characteristic properties commonly observed in virialized self-gravitating systems, such as galaxies and interstellar molecular clouds. We study the origin of these properties using a one-dimensional ring model that we propose in this paper. In this simple model, N particles are moving, on a circular ring fixed in three dimensional space, with mutual interaction of gravity. This model is suitable for the accurate symplectic integration method by which we argue the phase transition in this system. Especially, in between the extended phase and the collapsed phase, we find an interesting phase (halo phase) that has negative specific heat at the intermediate energy scale. Moreover, in this phase, there appear scaling properties and nonthermal and non-Gaussian velocity distributions. In contrast, these peculiar properties are never observed in other gas and core phases. Particles in each phase have a typical time scale of motions determined by the cutoff length xi, the ring radius R, and the total energy E. Thus all relaxation patterns of the system are determined by these three time scales. PMID- 11736041 TI - Non-Gaussian equilibrium in a long-range Hamiltonian system. AB - We study the dynamics of a system of N classical spins with infinite-range interaction. We show that, if the thermodynamic limit is taken before the infinite-time limit, the system does not relax to the Boltzmann-Gibbs equilibrium, but exhibits different equilibrium properties, characterized by stable non-Gaussian velocity distributions, Levy walks, and dynamical correlation in phase space. PMID- 11736042 TI - XY model in small-world networks. AB - The phase transition in the XY model on one-dimensional small-world networks is investigated by means of Monte Carlo simulations. It is found that long-range order is present at finite temperatures, even for very small values of the rewiring probability, suggesting a finite-temperature transition for any nonzero rewiring probability. Nature of the phase transition is discussed in comparison with the globally coupled XY model. PMID- 11736043 TI - Mean-field approximation for a limit order driven market model. AB - A mean-field variant of the model of limit order driven market introduced recently by Maslov is formulated and solved. The agents do not have any strategies and the memory of the system is kept within the order book. We show that the evolution of the order book is governed by a matrix multiplicative process. The resulting stationary distribution of step-to-step price changes is calculated. It exhibits a power-law tail with exponent 2. We obtain also the price autocorrelation function, which agrees qualitatively with the experimentally observed negative autocorrelation for short times. PMID- 11736044 TI - Near-critical confined fluids and Ising films: density-matrix renormalization group study. AB - Two-dimensional Ising strips subject to identical surface fields h(1)=h(2) > or =0 are studied for temperatures above and below the bulk critical temperature T(c) and a range of bulk fields h by means of the density-matrix renormalization group method. In the case of nonvanishing surface fields, the near-critical behavior of the solvation force f(solv), total adsorption Gamma, inverse longitudinal correlation length xi(parallel)( -1) and specific heat C(H) is strongly influenced by the (pseudo) capillary condensation that occurs below T(c). We obtain scaling functions of f(solv), Gamma, and xi(parallel)(-1). C(H) exhibits a weakly rounded singularity on crossing the pseudocoexistence line. We contrast these results with those for the case of free boundaries where, for temperatures slightly below T(c), f(solv) and C(H) exhibit a sharp extremum away from h=0. Our results have direct repercussions for the properties of near critical Ising films in three dimensions and we argue that the long-ranged solvation (Casimir) force in confined fluids should be more attractive in the neighborhood of the capillary critical point than exactly at the bulk critical point. PMID- 11736045 TI - Continuum time limit and stationary states of the minority game. AB - We discuss in detail the derivation of stochastic differential equations for the continuum time limit of the minority game. We show that all properties of the minority game can be understood by a careful theoretical analysis of such equations. In particular, (i) we confirm that the stationary state properties are given by the ground state configurations of a disordered (soft) spin system, (ii) we derive the full stationary state distribution, (iii) we characterize the dependence on initial conditions in the symmetric phase, and (iv) we clarify the behavior of the system as a function of the learning rate. This leaves us with a complete and coherent picture of the collective behavior of the minority game. Strikingly we find that the temperaturelike parameter, which is introduced in the choice behavior of individual agents turns out to play the role, at the collective level, of the inverse of a thermodynamic temperature. PMID- 11736046 TI - Kinetics of phase separation in ternary mixtures. AB - We present detailed results from Monte Carlo simulations of the kinetics of phase separation in ternary mixtures. We focus on the case of ABV mixtures (where V denotes a vacancy) and investigate segregation kinetics resulting from V-mediated dynamics. We provide heuristic arguments for the existence of different morphologies in various parameter regimes. Furthermore, we present comprehensive numerical results for various characteristic features of the domain growth process, e.g., real-space correlation functions, domain-size distribution functions, and growth laws. PMID- 11736047 TI - Nonequilibrium dynamics in the complex Ginzburg-Landau equation. AB - Results from a comprehensive analytical and numerical study of nonequilibrium dynamics in the two-dimensional complex Ginzburg-Landau equation have been presented. In particular, spiral defects have been used to characterize the domain growth law and the evolution morphology. An asymptotic analysis of the single-spiral correlation function shows a sequence of singularities-analogous to those seen for time-dependent Ginzburg-Landau models with O(n) symmetry, where n is even. PMID- 11736048 TI - Continuous model for vocal fold oscillations to study the effect of feedback. AB - In this work we study the effects of delayed feedback on vocal fold dynamics. To perform this study, we work with a vocal fold model that is made as simple as possible while retaining the spectral content characteristic of human vocal production. Our results indicate that, even with the simplest explanation for vocal fold oscillation, delayed feedback due to reflected sound in the vocal tract can lead to extremely rich dynamics. PMID- 11736049 TI - Two-dimensional model of a reaction-diffusion system as a typewriter. AB - Pattern formation is a common phenomenon, which appears in biological systems, especially in cell differentiation processes. The proper level for understanding the creation of patterns seems to be a physicochemical description. The most fundamental models should be based on systems, in which only chemical reactions and diffusion transport occur (reaction-diffusion systems). In order to present a richness of patterns, we show here the asymptotic patterns in the form of capital letters obtained in two-dimensional reaction-diffusion systems with zero-flux boundary conditions. All capital letters are obtained in the same model, but initial conditions and sizes of the systems are different for each letter. The chemical model consists of elementary reactions and is realistic. It can be realized experimentally in continuous-flow unstirred reactor with an enzymatic reaction allosterically inhibited by an excess of its reactant and product. PMID- 11736050 TI - Interplays between Harper and Mathieu equations. AB - This paper deals with the application of relationships between Harper and Mathieu equations to the derivation of energy formulas. Establishing suitable matching conditions, one proceeds by inserting a concrete solution to the Mathieu equation into the Harper equation. For this purpose, one resorts to the nonlinear oscillations characterizing the Mathieu equation. This leads to the derivation of two kinds of energy formulas working in terms of cubic and quadratic algebraic equations, respectively. Combining such results yields quadratic equations to the energy description of the Harper equation, incorporating all parameters needed. PMID- 11736051 TI - Unnested islands of period doublings in an injected semiconductor laser. AB - We present a theoretical study of unnested period-doubling islands in three dimensional rate equations modeling a semiconductor laser subject to external optical injection. In this phenomenon successive curves of period doublings are not arranged in nicely nested islands, but intersect each other. This overall structure is globally organized by several codimension-2 bifurcations. As a consequence, the chaotic region existing inside an unnested island of period doublings can be entered not only via a period-doubling cascade but also via the breakup of a torus, and even via the sudden appearance of a chaotic attractor. In order to fully understand these different chaotic transitions we reveal underlying global bifurcations and we show how they are connected to codimension 2 bifurcation points. Unnested islands of period doublings appear to be generic and hence must be expected in a large class of dynamical systems. PMID- 11736052 TI - Angular transport in a nonperiodic Chirikov-Taylor map. AB - Transport in angular direction is considered for a nonperiodic Chirikov-Taylor (standard) map. In the limit of large stochasticity parameter, depending on the boundary conditions of the action variable, either superdiffusive of diffusive behavior is found. In both cases characteristic oscillations in the transport coefficients occur. Theoretical predictions based on the Perron-Frobenius evolution operator formalism for the distribution function are compared with numerical simulations. Information on the anomalous behaviors in the near threshold as well as in the subthreshold regions are also presented. PMID- 11736053 TI - Limitation on stabilizing plane waves via time-delay feedback. AB - Previous work has demonstrated the possibility of stabilizing plane wave solutions of one-dimensional systems using a spatially local form of time-delayed feedback. We show that the natural extension of this method to two-dimensional systems fails due to the presence of torsion-free unstable perturbations. Linear stability analysis of the complex Ginzburg-Landau equation reveals that long wavelength, transverse wave instabilities cannot be suppressed by the method of extended time-delay autosynchronization. The conclusion follows from symmetry considerations and therefore applies to a wide class of models with simple plane wave solutions. PMID- 11736054 TI - Analyses of transient chaotic time series. AB - We address the calculation of correlation dimension, the estimation of Lyapunov exponents, and the detection of unstable periodic orbits, from transient chaotic time series. Theoretical arguments and numerical experiments show that the Grassberger-Procaccia algorithm can be used to estimate the dimension of an underlying chaotic saddle from an ensemble of chaotic transients. We also demonstrate that Lyapunov exponents can be estimated by computing the rates of separation of neighboring phase-space states constructed from each transient time series in an ensemble. Numerical experiments utilizing the statistics of recurrence times demonstrate that unstable periodic orbits of low periods can be extracted even when noise is present. In addition, we test the scaling law for the probability of finding periodic orbits. The scaling law implies that unstable periodic orbits of high period are unlikely to be detected from transient chaotic time series. PMID- 11736055 TI - Wave function statistics for ballistic quantum transport through chaotic open billiards: statistical crossover and coexistence of regular and chaotic waves. AB - For ballistic transport through chaotic open billiards, we implement accurate fully quantal calculations of the probability distributions and spatial correlations of the local densities of single-electron wave functions within the cavity. We find wave-statistical behaviors intrinsically different from those in their closed counterparts. Chaotic-scattering wave functions in open systems can be quantitatively interpreted in terms of statistically independent real and imaginary random fields in the same way as for wave-function statistics of closed systems in the time-reversal symmetry-breaking crossover regime. We also discuss perceived statistical deviations, which are attributed to the coexistence of regular and chaotic waves and given analytical explanations. PMID- 11736056 TI - Chaos and the continuum limit in the gravitational N-body problem: integrable potentials. AB - This paper summarizes a numerical investigation of the statistical properties of orbits evolved in "frozen," time-independent N-body realizations of smooth, time independent density distributions corresponding to integrable potentials, allowing for 10(2.5) < or = N < or = 10(5.5). Two principal conclusions were reached: (1) In agreement with recent work by Valluri and Merritt, one finds that, in the limit of a nearly "unsoftened" two-body kernel, i.e., V(r) approximately equals (r(2) + epsilon(2))(-1/2) for epsilon --> 0, the value of the largest Lyapunov exponent chi does not decrease systematically with increasing N, so that, viewed in terms of the sensitivity of individual orbits to small changes in initial conditions, there is no sense in which chaos "turns off" for large N. However, it is clear that, for any finite epsilon, chi will tend to zero for sufficiently large N. (2) Even though chi does not decrease for an unsoftened kernel, there is a clear, quantifiable sense in which, as N increases, chaotic orbits in the frozen-N systems remain "close to" integrable characteristics in the smooth potential for progressively longer times. When viewed in configuration or velocity space, or as probed by collisionless invariants like angular momentum, frozen-N orbits typically diverge from smooth potential characteristics as a power law in time, rather than exponentially, on a time scale approximately equals N(p)t(D), with p approximately 1/2 and t(D) a characteristic dynamical, or crossing, time. For the case of angular momentum, the divergence is well approximated by a t(1/2) dependence, so that, when viewed in terms of collisionless invariants, discreteness effects act as a diffusion process that, presumably, can be modeled by nearly white Gaussian noise in the context of a Langevin or Fokker-Planck description. For position and velocity, the divergence is more rapid, characterized by a nearly linear power-law growth, t(q) with q approximately 1, a result that likely reflects the effects of linear phase mixing. The inference that, pointwise, individual N-body orbits can be reasonably approximated by orbits in a smooth potential only for times < N(1/2)t(D) has potential implications for various resonance phenomena that can act in real self-gravitating systems. PMID- 11736057 TI - Selective communication and information processing by excitable systems. AB - The phenomena of selective response of an excitable system to external pulse stimulation relating to interneuron communication and information processing problems are discussed. Subthreshold dynamics of the FitzHugh-Nagumo-like excitable system modeling of a neuron with the synaptic input is investigated. It is shown that the system response on various incoming information messages can be described by one- and two-dimensional linear and nonlinear point maps. Nonlinear integrating and resonant properties of the system are analyzed. PMID- 11736058 TI - Nonhyperbolic escape and changes in phase-space stability structures in laser induced multiphoton dissociation of a diatomic molecule. AB - The dependence of photodissociation of a diatomic molecule (vibrating according to the Morse potential) on the frequency of the laser field that induces it, is studied in the context of classical nonlinear dynamics. First, it is observed that as the laser frequency increases towards the harmonic frequency of the potential, a transition from stabilization due to Kolmogorov-Arnold-Moser tori to stabilization caused by the resonance stability island occurs. Then, considering the photodissociation as a nonhyperbolic half-scattering process, we investigate the influence of these changes in the phase space stability structures on dissociation dynamics via the examination of the fractal set of singularities appearing in the time-delay function of the initial state. It is found that the effective fractal dimension of this set (a finite-scale approximation of the exact dimension which is always equal to 1) and the percentage of its singularities provide a link between these changes and the dissociation rate. PMID- 11736059 TI - Control of hyperchaos. AB - A general method for controlling chaotic systems with one or more positive Lyapunov exponents is investigated analytically and numerically. The method retains the formal features of the adaptive adjustment mechanism and can be equally applied to various types of the unstable fixed points. It is shown that the method proposed here neither asks for any prior analytical knowledge of the system, nor any internal or external controlling parameters in advance. PMID- 11736060 TI - Stability of Turing patterns in the Brusselator model. AB - The selection and competition of Turing patterns in the Brusselator model are reviewed. The stability of stripes and hexagons towards spatial perturbations is studied using the amplitude equation formalism. For hexagonal patterns these equations include both linear and nonpotential spatial terms enabling distorted solutions. The latter modify substantially the stability diagrams and select patterns with wave numbers quite different from the critical value. The analytical results from the amplitude formalism agree with direct simulations of the model. Moreover, we show that slightly squeezed hexagons are locally stable in a full range of distortion angles. The stability regions resulting from the phase equation are similar to those obtained numerically by other authors and to those observed in experiments. PMID- 11736061 TI - Whispering gallery modes in open quantum billiards. AB - The poles of the S matrix and the wave functions of open two-dimensional quantum billiards with convex boundary of different shape are calculated by using the method of complex scaling. Two leads are attached to the cavities. The conductance of the cavities is calculated at energies with one, two, and three open channels in each lead. Bands of overlapping resonance states appear that are localized along the convex boundary of the cavities and contribute coherently to the conductance. These bands correspond to the whispering gallery modes known from classical calculations. PMID- 11736062 TI - Classical nonlinearity and quantum decay: the effect of classical phase-space structures. AB - We investigate the decay process from a time dependent potential well in the semiclassical regime. The classical dynamics is chaotic and the decay rate shows an irregular behavior as a function of the system parameters. By studying the weak-chaos regime we are able to connect the decay irregularities to the presence of nonlinear resonances in the classical phase space. A quantitative analytical prediction that accounts for the numerical results is obtained. PMID- 11736063 TI - Reconstruction of time-delay systems from chaotic time series. AB - We propose a method that allows one to estimate the parameters of model scalar time-delay differential equations from time series. The method is based on a statistical analysis of time intervals between extrema in the time series. We verify our method by using it for the reconstruction of time-delay differential equations from their chaotic solutions and for modeling experimental systems with delay-induced dynamics from their chaotic time series. PMID- 11736064 TI - Variational approach to a class of nonlinear oscillators with several limit cycles. AB - We study limit cycles of nonlinear oscillators described by the equation x + nuF(x) + x = 0 with F an odd function. Depending on the nonlinearity, this equation may exhibit one or more limit cycles. We show that limit cycles correspond to relative extrema of a certain functional. Analytical results in the limits nu-->0 and nu-->infinity are in agreement with previously known criteria. For intermediate nu, numerical determination of the limit cycles can be obtained. PMID- 11736065 TI - Transition from oscillatory to excitable regime in a system forced at three times its natural frequency. AB - The effect of a temporal modulation at three times the critical frequency on a Hopf bifurcation is studied in the framework of amplitude equations. We consider a complex Ginzburg-Landau equation with an extra quadratic term, resulting from the strong coupling between the external field and the unstable modes. We show that, by increasing the intensity of the forcing, one passes from an oscillatory regime to an excitable one with three equivalent frequency-locked states. In the oscillatory regime, topological defects are one-armed phase spirals, while in the excitable regime they correspond to three-armed excitable amplitude spirals. Analytical results show that the transition between these two regimes occurs at a critical value of the forcing intensity. The transition between phase and amplitude spirals is confirmed by numerical analysis and it might be observed in periodically forced reaction-diffusion systems. PMID- 11736066 TI - Numerical analysis of spectra of the Frobenius-Perron operator of a noisy one dimensional mapping: toward a theory of stochastic bifurcations. AB - A different method to detect the stochastic bifurcation point of a one dimensional mapping in the presence of noise is proposed. This method analyzes the eigenvalues and eigenfunctions of the noisy Frobenius-Perron operator. The invariant density or the eigenfunction of the eigenvalue 1 of the operator possesses "static" information of the noisy one-dimensional dynamics while the other eigenvalues and eigenfunctions have "dynamic" information. Clear bifurcation phenomena have been observed in a noisy sine-circle map and both stochastic saddle-node and period-doubling bifurcation points have been successfully defined in terms of the eigenvalues. PMID- 11736067 TI - Transforming chaos to periodic oscillations. AB - We demonstrate that the dynamics of an autonomous chaotic class C laser can be controlled to a periodic state via external modulation of the pump. In the absence of modulation, above the chaos threshold, the laser exhibits Lorenz-like chaotic pulsations. The average amplitude and frequency of these pulsations depend on the pump power. We find that there exist parameter windows where modulation of the pump power extinguishes the chaos in favor of simpler periodic behavior. Moreover we find a number of locking ratios between the pump and laser output follow the Farey sequence. PMID- 11736068 TI - Effect of evanescent modes and chaos on deterministic scattering in electron waveguides. AB - Statistical properties of Wigner delay times and the effect of evanescent modes on the deterministic scattering of an electron matter wave from a classically chaotic two-dimensional electron waveguide are studied for the case of 2, 6, and 16 propagating modes. Deterministic reaction matrix theory for this system is generalized to include the effect of evanescent modes on the scattering process. The statistical properties of the Wigner delay times for the deterministic scattering process are compared to the predictions of random reaction matrix theory. PMID- 11736069 TI - Parameter estimation in spatially extended systems: the Karhunen-Loeve and Galerkin multiple shooting approach. AB - Parameter estimation for spatiotemporal dynamics for coupled map lattices and continuous time domain systems is shown using a combination of multiple shooting, Karhunen-Loeve decomposition and Galerkin's projection methodologies. The resulting advantages in estimating parameters have been studied and discussed for chaotic and turbulent dynamics using small amounts of data from subsystems, availability of only scalar and noisy time series data, effects of space-time parameter variations, and in the presence of multiple time scales. PMID- 11736070 TI - Semiclassical quantization of strongly chaotic vibrations in an M7-like cluster. AB - We report semiclassical energy spectra of vibrational state of a cluster composed of seven identical atoms like Ar7 in terms of our previously developed semiclassical wave function, which we call the action-decomposed function. The classical dynamics of this vibrational state is strongly chaotic and undergoes a large amplitude motion due to structural isomerization, which demands a long run of trajectory calculation. Permutation of identical particles should also be taken into account as a quantum effect, since a single molecular shape can be shared by many permutational isomers. Furthermore, chaos causes a spurious divergence in the amplitude factor of a correlation function in the initial value representation, which arises from the amplitude factor (prefactor) of a semiclassical wave function, while the final-state representation is suffered from the well-known divergence arising at caustics. Both approaches therefore face tremendous difficulty in a long-time calculation of the correlation functions. We challenge to extract some limited number of spectral lines from such chaotic dynamics. We further apply a correlation function that is free of such a troublesome amplitude factor. Numerical results from all these schemes are reported. PMID- 11736071 TI - Modeling of an impact system with a drift. AB - A physical model to examine impact oscillators has been developed and analyzed. The model accounts for the viscoelastic impacts and is capable to mimic the dynamics of a bounded progressive motion (a drift), which is important in practical applications. The system moves forward in stick-slip phases, and its behavior may vary from periodic to chaotic motion. A nonlinear dynamic analysis reveals a complex behavior and that the largest drift is achieved when the responses switch from periodic to chaotic, after a cascade of subcritical bifurcations to period one. Based on this fact, a semianalytical solution is constructed to calculate the progression of the system for periodic regimes and to determine conditions when periodicity is lost. PMID- 11736072 TI - Periodic forcing in viscous fingering of a nematic liquid crystal. AB - Viscous fingering of an air-nematic interface in a radial Hele-Shaw cell is studied when periodically switching on and off an electric field, which reorients the nematic and thus changes its viscosity, as well as the surface tension and its anisotropy (mainly enforced by a single groove in the cell). Undulations at the sides of the fingers are observed that correlate with the switching frequency and with tip oscillations that give maximal velocity to smallest curvatures. These lateral undulations appear to be decoupled from spontaneous (noise induced) side branching. It is concluded that the lateral undulations are generated by successive relaxations between two limiting finger widths. The change between these two selected pattern scales is mainly due to the change in the anisotropy. This scenario is confirmed by numerical simulations in the channel geometry, using a phase-field model for anisotropic viscous fingering. PMID- 11736073 TI - Delocalization border and onset of chaos in a model of quantum computation. AB - We study the properties of spectra and eigenfunctions for a chain of 1/2 spins (qubits) in an external time-dependent magnetic field and under the conditions of nonselective excitation (when the amplitude of the magnetic field is large). This model is known as a possible candidate for experimental realization of quantum computation. We present the theory for finding delocalization transitions and show that for the interaction between nearest qubits, the transition is very different from that in quantum chaos. We explain this phenomena by showing that in the considered region of parameters our model is close to an integrable one. According to a general opinion, the threshold for the onset of quantum chaos due to the interqubit interaction decreases with an increase of the number of qubits. Contrary to this expectation, for a magnetic field with constant gradient we have found that chaos border does not depend on the number of qubits. We give analytical estimates that explain this effect, together with numerical data supporting our analysis. Random models with long-range interactions have been studied as well. In particular, we show that in this case the delocalization and quantum chaos borders coincide. PMID- 11736074 TI - Effective coupling for open billiards. AB - We derive an explicit expression for the coupling constants of individual eigenstates of a closed billiard that is opened by attaching a waveguide. The Wigner time delay and the resonance positions resulting from the coupling constants are compared to an exact numerical calculation. Deviations can be attributed to evanescent modes in the waveguide and to the finite number of eigenstates taken into account. The influence of the shape of the billiard and of the boundary conditions at the mouth of the waveguide are also discussed. Finally we show that the mean value of the dimensionless coupling constants tends to the critical value when the eigenstates of the billiard follow random-matrix theory. PMID- 11736075 TI - Catastrophic bifurcation from riddled to fractal basins. AB - Most existing works on riddling assume that the underlying dynamical system possesses an invariant subspace that usually results from a symmetry. In realistic applications of chaotic systems, however, there exists no perfect symmetry. The aim of this paper is to examine the consequences of symmetry breaking on riddling. In particular, we consider smooth deterministic perturbations that destroy the existence of invariant subspace, and identify, as a symmetry-breaking parameter is increased from zero, two distinct bifurcations. In the first case, the chaotic attractor in the invariant subspace is transversely stable so that the basin is riddled. We find that a bifurcation from riddled to fractal basins can occur in the sense that an arbitrarily small amount of symmetry breaking can replace the riddled basin by fractal basins. We call this a catastrophe of riddling. In the second case, where the chaotic attractor in the invariant subspace is transversely unstable so that there is no riddling in the unperturbed system, the presence of a symmetry breaking, no matter how small, can immediately create fractal basins in the vicinity of the original invariant subspace. This is a smooth-fractal basin boundary metamorphosis. We analyze the dynamical mechanisms for both catastrophes of riddling and basin boundary metamorphoses, derive scaling laws to characterize the fractal basins induced by symmetry breaking, and provide numerical confirmations. The main implication of our results is that while riddling is robust against perturbations that preserve the system symmetry, riddled basins of chaotic attractors in the invariant subspace, on which most existing works are focused, are structurally unstable against symmetry-breaking perturbations. PMID- 11736076 TI - Breathing current domains in globally coupled electrochemical systems: a comparison with a semiconductor model. AB - Spatio-temporal bifurcations and complex dynamics in globally coupled intrinsically bistable electrochemical systems with an S-shaped current-voltage characteristic under galvanostatic control are studied theoretically on a one dimensional domain. The results are compared with the dynamics and the bifurcation scenarios occurring in a closely related model which describes pattern formation in semiconductors. Under galvanostatic control both systems are unstable with respect to the formation of stationary large amplitude current domains. The current domains as well as the homogeneous steady state exhibit oscillatory instabilities for slow dynamics of the potential drop across the double layer, or across the semiconductor device, respectively. The interplay of the different instabilities leads to complex spatio-temporal behavior. We find breathing current domains and chaotic spatio-temporal dynamics in the electrochemical system. Comparing these findings with the results obtained earlier for the semiconductor system, we outline bifurcation scenarios leading to complex dynamics in globally coupled bistable systems with subcritical spatial bifurcations. PMID- 11736077 TI - Energetics of interacting magnetized domains. AB - Many of the pattern forming features of ferrofluids, lipid monolayers, type-I superconductors, and magnetic bubbles can be understood by treating them as dipolar (uniformly magnetized or polarized) domains. Here, we investigate the early stages of pattern formation in a system consisting of two quasi-two dimensional dipolar domains. We calculate the linearized interaction energy for these domains and find that the lowest energy states are those for which each drop has an indentation directed toward the other. These theoretically preferred states compare favorably with results of experiments performed with ferrofluids. PMID- 11736078 TI - Phase-locked spatial domains and Bloch domain walls in type-II optical parametric oscillators. AB - We study the role of transverse spatial degrees of freedom in the dynamics of signal-idler phase locked states in type-II optical parametric oscillators. Phase locking stems from signal-idler polarization coupling which arises if the cavity birefringence and/or dichroism is not matched to the nonlinear crystal birefringence. Spontaneous Bloch domain wall formation is observed numerically and the dynamics and chiral properties of the fronts are investigated. Bloch walls connect homogeneous regions of self-phase-locked solutions by means of a polarization transformation. The parameter range for phase locking is found analytically. The polarization properties and the dynamics of walls in one and two transverse spatial dimensions are explained. The transition from Bloch to Ising walls is characterized, the control parameter being the linear coupling strength. The wall dynamics governs spatiotemporal dynamical states of the system, which include transient curvature driven domain growth, persistent dynamics dominated by spiraling defects for Bloch walls, and labyrinthine pattern formation for Ising walls. PMID- 11736079 TI - Liouvillian dynamics of the Hopf bifurcation. AB - Two-dimensional vector fields undergoing a Hopf bifurcation are studied in a Liouville-equation approach. The Liouville equation rules the time evolution of statistical ensembles of trajectories issued from random initial conditions, but evolving under the deterministic dynamics. The time evolution of the probability densities of such statistical ensembles can be decomposed in terms of the spectrum of the resonances (i.e., the relaxation rates) of the Liouvillian operator or the related Frobenius-Perron operator. The spectral decomposition of the Liouvillian operator is explicitly constructed before, at, and after the Hopf bifurcation. Because of the emergence of time oscillations near the Hopf bifurcation, the resonance spectrum turns out to be complex and defined by both relaxation rates and oscillation frequencies. The resonance spectrum is discrete far from the bifurcation and becomes continuous at the bifurcation. This continuous spectrum is caused by the critical slowing down of the oscillations occurring at the Hopf bifurcation and it leads to power-law relaxation as 1/square root of [t] of the probability densities and statistical averages at long times t-->infinity. Moreover, degeneracy in the resonance spectrum is shown to yield a Jordan-block structure in the spectral decomposition. PMID- 11736080 TI - Approaching classicality in quantum accelerator modes through decoherence. AB - We describe measurements of the mean energy of an ensemble of laser-cooled atoms in an atom optical system in which the cold atoms, falling freely under gravity, receive approximate delta-kicks from a pulsed standing wave of laser light. We call this system a "delta-kicked accelerator." Additionally, we can counteract the effect of gravity by appropriate shifting of the position of the standing wave, which restores the dynamics of the standard delta-kicked rotor. The presence of gravity (delta-kicked accelerator) yields quantum phenomena, quantum accelerator modes, which are markedly different from those in the case for which gravity is absent (delta-kicked rotor). Quantum accelerator modes result in a much higher rate of increase in the mean energy of the system than is found in its classical analog. When gravity is counteracted, the system exhibits the suppression of the momentum diffusion characteristic of dynamical localization. The effect of noise is examined and a comparison is made with simulations of both quantum-mechanical and classical versions of the system. We find that the introduction of noise results in the restoration of several signatures of classical behavior, although significant quantum features remain. PMID- 11736081 TI - Phase synchronization of chaotic systems with small phase diffusion. AB - The geometric theory of phase locking between periodic oscillators is extended to phase coherent chaotic systems. This approach explains the qualitative features of phase locked chaotic systems and provides an analytical tool for a quantitative description of the phase locked states. Moreover, this geometric viewpoint allows us to identify obstructions to phase locking even in systems with negligible phase diffusion, and to provide sufficient conditions for phase locking to occur. We apply these techniques to the Rossler system and a phase coherent electronic circuit and find that numerical results and experiments agree well with theoretical predictions. PMID- 11736082 TI - Symmetry-increasing bifurcation as a predictor of a chaos-hyperchaos transition in coupled systems. AB - In weakly coupled systems, it is possible to observe the coexistence of the chaotic attractors which are located out of the invariant manifold and are not symmetrical in relation to this manifold. When the control parameter is changed, these attractors can undergo a chaos-hyperchaos transition. We give numerical evidence that before this transition the coexisting attractors merge together creating an attractor symmetrical with respect to the invariant manifold. We argue that the attractors that are not located at the invariant manifold can exhibit dynamical behavior similar to bubbling and on-off intermittency previously observed for the attractors located at the invariant manifold, and we describe the mechanism of these phenomena. PMID- 11736083 TI - Dynamical mechanisms of dc current generation in driven Hamiltonian systems. AB - Recent symmetry considerations [Flach et al., Phys. Rev. Lett. 84, 2358 (2000)] have shown that dc currents may be generated in the stochastic layer of a system describing the motion of a particle in a one-dimensional potential in the presence of an ac time-periodic drive. In this paper we explain the dynamical origin of this current. We show that the dc current is induced by the presence and desymmetrization of ballistic channels inside the stochastic layer. The existence of these channels is due to resonance islands with nonzero winding numbers. The characterization of the flight dynamics inside ballistic channels is described by distribution functions. We obtain these distribution functions numerically and find very good agreement with simulation data. PMID- 11736084 TI - Electronic states in a cylindrical quantum lens: quantum chaos for decreasing system symmetry. AB - The eigenvalue problem in a cylindrical lens geometry is studied. Using a conformal mapping method, the shape of the boundary and the Hamiltonian for a free particle are reduced to those of a two-dimensional problem with circular symmetry. The wave functions are separated into two independent Hilbert subspaces due to the inherent symmetry of the problem. For small geometry deformations, the solutions are found by a specially designed perturbation approach. Comparisons between exact and perturbative solutions are made for different lens parameters. As the symmetry of the lens is reduced, the characteristics of the spectrum and the corresponding spatial properties of the wave functions are studied. Our results provide a family of billiard geometries in which the electronic level spectrum is well characterized. In analyzing the level spacing distribution of the spectrum, a strong deviation from the Poisson and Wigner limiting distributions is found as the boundary geometry changes. This intermediate distribution is indicative of a mixed phase space, also revealed explicitly in the classical Poincare maps we present. PMID- 11736085 TI - Decoherence in a classically chaotic quantum system: entropy production and quantum-classical correspondence. AB - We study the decoherence process for an open quantum system that is classically chaotic (a quartic double well with harmonic driving coupled to a sea of harmonic oscillators). We carefully analyze the time dependence of the rate of entropy production showing that it has two relevant regimes: For short times it is proportional to the diffusion coefficient (fixed by the system-environment coupling strength); for longer times (but before equilibration) it is fixed by dynamical properties of the system (and is related to the Lyapunov exponent). The nature of the transition time between both regimes is investigated and the issue of quantum to classical correspondence is addressed. Finally, the impact of the interaction with the environment on coherent tunneling is analyzed. PMID- 11736086 TI - Information circulation in a two-mode solid-state laser with optical feedback. AB - A two-mode solid-state laser subjected to a delayed optical feedback is studied. Simultaneous random switchings between stable and chaotic antiphase spiking oscillations featuring the establishment of causal (drive response) relationships among modes have been demonstrated by a proposed information circulation analysis of an experimental time series. The observed phenomenon has been well reproduced by numerical simulations of two-mode laser equations with uncorrelated modal phase fluctuations. PMID- 11736087 TI - Energy fluctuations of pseudointegrable systems with growing surface roughness. AB - The eigenfrequencies of two-dimensional systems with fractal boundaries and with nonscaling rough boundaries are calculated numerically by the Lanczos algorithm and analyzed by means of level statistics. The systems are pseudointegrable and the fluctuations of their eigenvalue spectra show a global statistical behavior between the Poisson and the Wigner distributions. With increasing irregularity of the boundary, the systems approach the Wigner limit and the results seem to depend only on the genus number of the geometry and not on details, such as the asymptotic shape of the geometry, the type of roughness (scaling or nonscaling), and the boundary conditions (Neumann or Dirichlet). No transition between localized and extended states is found in fractal drums. PMID- 11736088 TI - Turbulence of polymer solutions. AB - We investigate high-Reynolds-number turbulence in dilute polymer solutions. We show the existence of a critical value of the Reynolds number, which separates two different regimes. In the first regime, below the transition, the influence of the polymer molecules on the flow is negligible, so they can be regarded as passively embedded in the flow. This case admits a detailed investigation of the statistics of the polymer elongations. The second state is realized when the Reynolds number is larger than the critical value. This regime is characterized by the strong back reaction of polymers on the flow. We establish some properties of the statistics of the stress and velocity in this regime and discuss its relation to the drag reduction phenomenon. PMID- 11736089 TI - Dynamical equations for high-order structure functions, and a comparison of a mean-field theory with experiments in three-dimensional turbulence. AB - Two recent papers [V. Yakhot, Phys. Rev. E 63, 026307, (2001) and R. J. Hill, J. Fluid Mech. 434, 379, (2001)] derive, through two different approaches that have the Navier-Stokes equations as the common starting point, a set of dynamic equations for structure functions of arbitrary order in turbulence. These equations are not closed. Yakhot proposed a "mean-field theory" to close the equations for locally isotropic turbulence, and obtained scaling exponents of structure functions and expressions for the peak in the probability density function of transverse velocity increments, and for its behavior for intermediate amplitudes. At high Reynolds numbers, some relevant experimental data on pressure gradient and dissipation terms are presented that are needed to provide closure, as well as on other aspects predicted by the theory. Comparison between the theory and the data shows varying levels of agreement, and reveals gaps inherent to the implementation of the theory. PMID- 11736090 TI - Evolution of triangles in a two-dimensional turbulent flow. AB - As a turbulent flow advects a swarm of Lagrangian markers, the mutual separation between particles grows, and the shape of the swarm gets distorted. By following three points in an experimental turbulent two-dimensional flow with a k(-5/3) spectrum, we investigate the geometry of triangles, in a statistical sense. Two well-characterized shape distributions are identified. At long times when the average size of the triangles is larger than the integral scale, the distribution of shapes is Gaussian. When the size of the triangle is in the inertial range and grows as t(3/2) (Richardson's law), a plausibly self-similar, non-Gaussian probability distribution is observed, where very elongated triangles have a much larger probability than in the Gaussian regime. These results are discussed, and, in the latter case, compared with the predictions of a stochastic model recently introduced [A. Pumir et al., Phys. Rev. Lett. 85, 5324 (2000)]. PMID- 11736091 TI - Self-similar fluctuation and large deviation statistics in the shell model of turbulence. AB - Both static and dynamic multiscalings of fluctuations of energy flux and energy dissipation rate in the Gledzer-Ohkitani-Yamada (GOY) shell model of turbulence are numerically investigated. We compute the large deviation rate function of energy flux not only in the inertial range (IR) but also around the crossover between the inertial range and the dissipation range (DR). The rate function in IR exists to be concave, which assures the applicability of the Legendre transformation with the anomalous scaling exponents that have been investigated so far, and turns out to be independent of the Reynolds number. On the contrary, near the crossover scale, an intermediate dissipation range (IMDR) scaling is observed with the rate function in IMDR, which is accounted with the argument on dissipation scale fluctuation dominated by the energy flux fluctuation in the inertial range. Furthermore, to study the difference between IR intermittency and DR intermittency, we compute finite time-averaged quantities of energy flux and energy dissipation rate and investigate their multiscaling behavior. The difference observed in terms of their dynamic multiscaling is discussed. PMID- 11736092 TI - Advective transport in the percolation backbone in two dimensions. AB - We show that, in the case of advective transport on the percolation backbone, the relevant structure below the correlation length is an ensemble of tortuous paths, rather than the classical links-nodes-blobs system. These paths are embedded in the few largest blobs that dominate the structure of the backbone. We find numerically that the mean particle displacement differs from the prediction given by classical finite-size scaling arguments. We also show that because of the complex velocity distribution between the paths, the mean first-passage time of the particles cannot be inferred directly from the mean particle displacement. PMID- 11736093 TI - Calculation of the anomalous exponents in the rapid-change model of passive scalar advection to order epsilon(3). AB - The field theoretic renormalization group and operator product expansion are applied to the model of a passive scalar advected by the Gaussian velocity field with zero mean and correlation function approximately equal to delta(t-t('))/k(d + epsilon). Inertial-range anomalous exponents, identified with the critical dimensions of various scalar and tensor composite operators constructed of the scalar gradients, are calculated within the epsilon expansion to order epsilon(3) (three-loop approximation), including the exponents in anisotropic sectors. The main goal of the paper is to give the complete derivation of this third-order result, and to present and explain in detail the corresponding calculational techniques. The character and convergence properties of the epsilon expansion are discussed, the improved "inverse" epsilon expansion is proposed, and the comparison with the existing nonperturbative results is given. PMID- 11736094 TI - Nonlinear turbulent magnetic diffusion and mean-field dynamo. AB - The nonlinear coefficients defining the mean electromotive force (i.e., the nonlinear turbulent magnetic diffusion, the nonlinear effective velocity, the nonlinear kappa tensor, etc.) are calculated for an anisotropic turbulence. A particular case of an anisotropic background turbulence (i.e., the turbulence with zero-mean magnetic field) with one preferential direction is considered. It is shown that the toroidal and poloidal magnetic fields have different nonlinear turbulent magnetic diffusion coefficients. It is demonstrated that even for a homogeneous turbulence there is a nonlinear effective velocity that exhibits diamagnetic or paramagnetic properties depending on the anisotropy of turbulence and the level of magnetic fluctuations in the background turbulence. The diamagnetic velocity results in the field being pushed out from the regions with stronger mean magnetic field, while the paramagnetic velocity causes the magnetic field to be concentrated in the regions with stronger field. Analysis shows that an anisotropy of turbulence strongly affects the nonlinear turbulent magnetic diffusion, the nonlinear effective velocity, and the nonlinear alpha effect. Two types of nonlinearities (algebraic and dynamic) are also discussed. The algebraic nonlinearity implies a nonlinear dependence of the mean electromotive force on the mean magnetic field. The dynamic nonlinearity is determined by a differential equation for the magnetic part of the alpha effect. It is shown that for the alphaOmega axisymmetric dynamo the algebraic nonlinearity alone (which includes the nonlinear alpha effect, the nonlinear turbulent magnetic diffusion, the nonlinear effective velocity, etc.) cannot saturate the dynamo generated mean magnetic field while the combined effect of the algebraic and dynamic nonlinearities limits the mean magnetic field growth. PMID- 11736095 TI - Theoretical study of opacity for a mixture of gold and gadolinium at a high temperature. AB - Using the detailed configuration accounting with the term structures treated by the unresolved transition array model, we have presented a method to calculate the spectral-resolved opacity for high temperature and density plasmas. Due to the fully relativistic treatment, incorporated with the quantum defect theory to handle the huge number of transition arrays from configurations with high principal quantum number, we can calculate the opacity of any medium- and high-Z plasmas conveniently. In the present work, the frequency-dependent opacity and the Rosseland mean opacity are calculated for a mixture of gold and gadolinium at a high temperature, 250 eV, and three densities, 0.1 g/cm(3), 1.0 g/cm(3), and 10.0 g/cm(3). Agreement between our theoretical results and experimental measurements and other theoretical simulations is obtained. PMID- 11736096 TI - Plasma hydrodynamics of the intense laser-cluster interaction. AB - We present a one-dimensional hydrodynamic model of the intense laser-cluster interaction in which the laser field is treated self-consistently. We find that for clusters initially as small as approximately 25 A in radius nonuniform expansion of the heated material results in long-time resonance of the laser field at the critical density plasma layer. A significant result of this is that the ponderomotive force, which is enhanced at the critical density surface, can be large enough to strongly modify the plasma hydrodynamics, even at laser intensities as low as 10(15) W/cm(2) for 800 nm laser pulses. Simulations of our recent experiments in extreme ultraviolet and x-ray generation from clusters explain the dependence of generation efficiency on laser pulse width. PMID- 11736097 TI - Electrical conductivity measurements of strongly coupled W plasmas. AB - Nonideal plasmas of tungsten were produced by vaporizing thin wires of 0.1 mm and 0.3 mm in diameter in small glass capillaries (in air) by means of a short-pulse current from an electrical discharge. For a short period of time, the inner wall of the rigid glass capillary confines the plasma until the induced pressure shock wave disintegrates the capillary. Spectroscopic measurements were carried out on the ejected plasma close to the end of the capillary. The plasma temperature was obtained by fitting a Planck function to the measured continuum spectrum. The resistance was derived from the voltage across the plasma and the current through the plasma. The plasma radius was determined with an intensified charge-coupled device camera and a streak camera and allowed the derivation of the conductivity. Particle densities were of the order of 10(22) particle/cm(3) and electron temperatures were in the range from 10 kK to 22 kK. These measurements are compared with theoretical models and previous work. PMID- 11736098 TI - Canonical Hamiltonian gyrocenter variables and gauge invariant representation of the gyrokinetic equation. AB - By using the extended phase space Hamiltonian Lie-transform perturbation method, Hamiltonian canonical variables have been found to describe the motion of gyrocenters. A representation of the gyrokinetic equation has been established in terms of the magnetic moment M, the total energy U, and the canonical toroidal momentum P of the particle. This representation of the gyrokinetic equation is invariant with respect to the gauge transformation of perturbation fields. It explicitly reveals the effects of toroidal symmetry breaking, and it indicates the role that the perturbed canonical toroidal momentum plays in the gyrokinetic theory. In particular, it is found that the free energy associated with partial differential(P)f(0)(M,U,P) [here f(0)(M,U,P) is the equilibrium distribution function] does not have any nonadiabatic linear driving to the axisymmetric modes. PMID- 11736099 TI - Inverse cascade in decaying three-dimensional magnetohydrodynamic turbulence. AB - We perform direct numerical simulations of three-dimensional freely decaying magnetohydrodynamic turbulence. For helical magnetic fields, an inverse cascade effect is observed in which power is transferred from smaller scales to larger scales. The magnetic field reaches a scaling regime with self-similar evolution, and power-law behavior at high wave numbers. We also find power-law decay in the magnetic and kinematic energies, and power-law growth in the characteristic length scale of the magnetic field. PMID- 11736100 TI - Transparency of an overdense plasma layer. AB - The transition from opacity to transparency of overdense plasma layers for the propagation of high-intensity laser pulses of circular polarization is investigated theoretically and by simulation. We discuss in detail the regions where stationary solutions exist and therefore the plasma layer is opaque. We find that the opacity depends not only on the electron density and the laser intensity but also on the thickness of the plasma layer. We also discuss the overlap region where both transparency and opacity are possible and how it depends on the history. Ion acceleration is studied in both the opacity region and the transparency region. PMID- 11736101 TI - Positively charged particles in dusty plasmas. AB - The trapping of dust particles has been observed in a dc abnormal glow discharge dominated by electron attachment. A dust cloud of several tens of positively charged particles was found to form in the anode sheath region. An analysis of the experimental conditions revealed that these particles were positively charged due to emission process, in contrast to most other experiments on the levitation of dust particles in gas-discharge plasmas where negatively charged particles are found. An estimate of the particle charge, taking into account the processes of photoelectron and secondary electron emission from the particle surface, is in agreement with the experimental measured values. PMID- 11736102 TI - Stochastic growth of ion cyclotron and mirror waves in Earth's magnetosheath. AB - Electromagnetic ion cyclotron and mirror waves in Earth's magnetosheath are bursty, have widely variable fields, and are unexpectedly persistent, properties difficult to reconcile with uniform secular growth. Here it is shown for specific periods that stochastic growth theory (SGT) quantitatively accounts for the functional form of the wave statistics and qualitatively explains the wave properties. The wave statistics are inconsistent with uniform secular growth or self-organized criticality, but nonlinear processes sometimes play a role at high fields. The results show SGT's relevance near marginal stability and suggest that it is widely relevant to space and astrophysical plasmas. PMID- 11736103 TI - Mass effects of light ion swarms in ac electric fields. AB - Transport properties of light ions in gases in ac electric fields are investigated by solving the time-dependent Boltzmann's equation. We focus on the way in which transport properties including diffusion are influenced by the ion neutral mass ratio as well as the field frequency. Calculations of transient relaxation phenomena in step-function fields are performed as an aid to understanding the complex temporal profiles of ion transport coefficients in ac electric fields. PMID- 11736104 TI - Dynamic structure factor for a two-component model plasma. AB - Analytical results for the structure factor of a two-component model plasma that describe an electron-ion plasma with modified interaction are derived from a Green function approach in different approximations. The random-phase approximation is improved by including the dynamic collision frequency, and results for the long-wavelength limit are extended to arbitrary wave numbers using the Mermin ansatz. After taking the classical limit of the resulting expressions, they are compared with molecular dynamics simulation results for the classical two-component model plasma. PMID- 11736105 TI - Stochastic magnetohydrodynamic turbulence in space dimensions d > or =2. AB - Interplay of kinematic and magnetic forcing in a model of a conducting fluid with randomly driven magnetohydrodynamic equations has been studied in space dimensions d > or =2 by means of the renormalization group. A perturbative expansion scheme, parameters of which are the deviation of the spatial dimension from two and the deviation of the exponent of the powerlike correlation function of random forcing from its critical value, has been used in one-loop approximation. Additional divergences have been taken into account that arise at two dimensions and have been inconsistently treated in earlier investigations of the model. It is shown that in spite of the additional divergences, the kinetic fixed point associated with the Kolmogorov scaling regime remains stable for all space dimensions d > or =2 for rapidly enough falling off correlations of the magnetic forcing. A scaling regime driven by thermal fluctuations of the velocity field has been identified and analyzed. The absence of a scaling regime near two dimensions driven by the fluctuations of the magnetic field has been confirmed. A renormalization scheme has been put forward and numerically investigated to interpolate between the epsilon expansion and the double expansion. PMID- 11736106 TI - Direct observation of the ponderomotive force effects in short-scale-length laser plasmas by frequency-domain interferometry. AB - We report on single-shot frequency-domain interferometric measurements showing space- and time-resolved ponderomotive electron density profile steepening of a short-scale-length ultraintense laser-produced plasma. The density gradient scale length is varied by applying a time-delayed laser prepulse. The measured absolute position of the critical density surface is found to be in agreement with one dimensional hydrodynamic simulations for the range of scale lengths studied. PMID- 11736107 TI - Radiation damping in real time. AB - We study the nonequilibrium dynamics of a charge interacting with its own radiation, which originates the radiation damping. The real-time equation of motion for the charge and the associated Langevin equation is found in classical limit. The equation of motion for the charge allows one to obtain the frequency dependent coefficient of friction. In the lowest order we find that although the coefficient of static friction vanishes, there is dynamical dissipation represented by a non-Markovian dissipative kernel. PMID- 11736108 TI - Quantum effects in high-gain free-electron lasers. AB - A many-particle fully quantized theory for a free-electron laser which is valid in the high-gain regime is presented. We examine quantum corrections for the high gain single-pass free-electron laser. It is shown that quantum effects become significant when the photon energy becomes comparable to the gain bandwidth. The initiation of the free-electron laser process from quantum fluctuations in the position and momentum of the electrons is considered, and the parameter regime for enhanced start-up is identified. Photon statistics of the free-electron laser radiation are discussed, and the photon number statistics for the self-amplified spontaneous emission are calculated. PMID- 11736109 TI - Wake field of an electron bunch moving parallel to a dielectric cylinder. AB - The wake field of an electron bunch moving parallel to the axis of a dielectric cylinder is being considered. It is shown that for a relativistic bunch (gamma>>1) the circular harmonic of order zero contributes a decelerating force inversely proportional to gamma, whereas the circular harmonics of nonzero order contribute a gamma-independent force. Moreover, the wake linked to the circular harmonic of order zero may grow in space in case the dielectric cylinder consists of an active medium; however, this growth rate does not depend on the value of gamma. On the other hand, no growth is anticipated for the case of circular harmonics of nonzero order. PMID- 11736110 TI - Generation of higher-order optical (2+1)-dimensional spatial vector solitons in a nonlinear anisotropic medium. AB - We investigate the generation of higher-order optical vector solitons in two transverse dimensions in anisotropic nonlinear media consisting of an incoherent superposition of a Gaussian beam and a higher-order laser mode with a complex internal modal structure. We demonstrate both numerically and experimentally various examples of these stable self-trapped light structures and show that vortex modes carrying topological charge always decay into multiple-humped structures that remain self trapped during propagation. Furthermore, we demonstrate the mutual stabilization of a triple- and a double-humped transverse light structure leading to the formation of a two-dimensional vector soliton without a stabilizing fundamental Gaussian mode. PMID- 11736111 TI - Construction of exact solutions by spatial translations in inhomogeneous nonlinear Schrodinger equations. AB - In this paper, we study a general nonlinear Schrodinger equation with a time dependent harmonic potential. Despite the lack of translational invariance, we find a symmetry transformation that, up from any solution, produces infinitely many others that are centered on classical trajectories. The results presented here imply that, not only the center of mass of the wave packet satisfies the Ehrenfest theorem and is decoupled from the dynamics of the wave packet, but also the shape of the solution is independent of the behavior of the center of the wave. Our findings have implications on the dynamics of Bose-Einstein condensates in magnetic traps. PMID- 11736112 TI - Incoherently coupled steady-state soliton pairs in biased photorefractive photovoltaic materials. AB - A theory on incoherently coupled soliton pairs for photorefractive screening solitons is developed that gives rise to incoherently coupled steady-state soliton pairs and multicomponent spatial solitons in biased photorefractive photovoltaic materials, which result from both the bulk photovoltaic effect and the spatially nonuniform screening of the external bias field. When the bulk photovoltaic effect is neglectable, these soliton pairs are the previously studied soliton pairs for screening solitons, these multicomponent spatial solitons predict incoherently coupled multicomponent spatial solitons for screening solitons, and their space-charge field is the space-charge field of screening solitons. When the external field is absent, these soliton pairs and multicomponent spatial solitons predict incoherently coupled soliton pairs and multicomponent spatial solitons for photovoltaic solitons, and their space-charge field is the space-charge field of photovoltaic solitons. The stability of these soliton pairs and multicomponent spatial solitons is also discussed using the modulation instability theory. PMID- 11736113 TI - Optical pulse propagation in nonlinear photonic crystals. AB - We present a formalism for optical pulse propagation in nonlinear photonic crystals of arbitrary dimensionality. Using a multiple-scale analysis, we derive the dynamical nonlinear Schrodinger equation obeyed by the envelope function modulating an underlying Bloch function. Effective coefficients appear in that equation characterizing the effects of Kerr nonlinearity, linear gain or loss, and material dispersion. They depend on how the underlying Bloch function "samples" these effects in the photonic crystal, and require for their calculation a specification of these effects throughout the photonic crystal, and the calculated bandstructure of the photonic crystals in the linear, nondispersive limit. We show that wave packets from different bands can experience significantly modified effective material properties. PMID- 11736114 TI - Solitary wave dynamics in generalized Hertz chains: an improved solution of the equation of motion. AB - The equation of motion for a bead in a chain of uncompressed elastic beads in contact that interact via the potential V(delta) approximately delta( n), n>2, delta being overlap, supports solitary waves and does not accommodate sound propagation [V. Nesterenko, J. Appl. Mech. Tech. Phys. 5, 733 (1983)]. We present an iteratively exact solution to describe the solitary wave as a function of material parameters and a universal, infinite set of coefficients, which depend only on n. We compute any arbitrary number of coefficients to desired accuracy and show that only the first few coefficients of our solution significantly improves upon Nesterenko's solution. The improved solution is a necessary step to develop a theoretical understanding of the formation of secondary solitary waves [M. Manciu, et al., Phys. Rev. E 63, 011614 (2001)]. PMID- 11736115 TI - Pattern formation and localization in the forced-damped Fermi-Pasta-Ulam lattice. AB - We study spatial pattern formation and energy localization in the dynamics of an anharmonic chain with quadratic and quartic intersite potential, subject to an optical, sinusoidally oscillating field and a weak damping. The zone-boundary mode is stable and locked to the driving field below a critical forcing that we determine analytically using an approximate model, which describes mode interactions. Above such a forcing, a standing modulated wave forms for driving frequencies below the band edge, while a "multibreather" state develops at higher frequencies. Of the former, we give an explicit approximate analytical expression, which compares well with numerical data. At higher forcing, space time chaotic patterns are observed. PMID- 11736116 TI - Acoustic pulse propagation and localization in bubbly water. AB - Acoustic pulse propagation in bubbly water is studied using a self-consistent method. The acoustic transmission and backscattering are evaluated numerically. Under proper conditions, the localization of acoustic waves is identified within a range of frequency. The results show that when a short pulse is transmitted the waves of frequencies within the localization regime will be trapped in the system and reveal a coherent behavior. A phase diagram approach is used to describe the localization behavior. PMID- 11736117 TI - Nonequivalence of phonon modes in the sine-Gordon equation. AB - We study the resonances in the sine-Gordon equation driven by an ac force using a linear perturbation theory. We show that resonances take place when the driving frequency delta is equal to half of the phonon modes' frequencies as has been shown numerically in our earlier work [N. R. Quintero, A. Sanchez, and F. G. Mertens, Phys. Rev. E 62, R60 (2000)], however, we find that the ac force is able to excite not all the phonon modes, but rather only the odd phonons (i.e., the ones with odd eigenfunctions). PMID- 11736118 TI - Electromagnetic properties of the linear oscillating currents flowing through toroidal knots. AB - Magnetic and toroid formfactors as well as the radiation intensity for a linear oscillating current flowing through a (knotted) toroidal spiral with a fractional number n=q/p(p,q=integers) of turns of winding are calculated. Some topological properties of such current lines are analyzed by studying the isotopic deformations when the ratio epsilon of the torus radii varies from 0 to 1 and a critical value is found for it, epsilon(c)=p(2)/(p(2)+q(2)), at which the Calugareanu invariant K (integer number) jumps by q units. PMID- 11736119 TI - Stability and formation of localized surface waves at the dielectric- photorefractive crystal boundary. AB - We consider specific features of the formation of localized surface waves at the interface between linear dielectric and photorefractive crystals with a nonlocal diffusion component of nonlinear response. Profiles of the surface waves are numerically found and guiding properties of the surface are investigated. Stability of the obtained surface waves is considered and it is shown that the well-known Vakhitov-Kolokolov stability criterion derived for the local Kerr or saturable material remains legible for the medium with a nonlocal diffusion component of nonlinear response. PMID- 11736120 TI - Microwave magnetoelectric particles: an experimental study of oscillating spectrums. AB - One of the ways to uncover the nature of the microwave magnetoelectric (ME) effect, recently observed in small ferrite resonators with special-form surface metallizations, is a comparative analysis of oscillating spectrums excited by different type rf external fields. Experimental results of the ME coupling in different types of ferrite resonators and different types of surface electrodes are reported and some important conclusions are drawn observing the oscillating spectrums of those particles. A special interest in spectral properties of point ME particles should be found in the field of microwave artificial composite materials-bianisotropic materials. PMID- 11736121 TI - Nonlinear dynamics of higher-order solitons near the oscillatory instability threshold. AB - Nonlinear theory describing the dynamics of solitons in the vicinity of oscillatory instability threshold with a low frequency offset is developed. The theory is tested on the example of parametric degenerate four-wave mixing. All major predictions of our theory are in agreement with the results of direct numerical modeling. This includes the position of oscillatory instability threshold, instability rates, and various instability development scenarios. PMID- 11736122 TI - Fractal structures and multiparticle effects in soliton scattering. AB - We study in detail the interaction of composite solitary waves and consider, as an example, the breather collisions in a weakly discrete sine-Gordon equation. We reveal a physical mechanism of fractal soliton scattering associated with multiparticle effects, and demonstrate chaotic interaction of two breathers with incommensurable frequencies. PMID- 11736123 TI - Transverse magnetic defect modes in two-dimensional triangular-lattice photonic crystals. AB - We present a numerical study of the localized transverse magnetic (TM) defect modes in a two-dimensional, triangular-lattice photonic crystal. The sample consists of an array of circular, air cylinders in a dielectric medium (GaAs). The defect modes were calculated by using a parallel version of the finite difference time-domain method on the Yee mesh. To validate our computations the results for the transverse electric case were checked against experimental results and the numerical results using a different method. We study the spatial symmetry for TM modes, obtained by changing the dipole excitation frequency. Also, we vary the defect-cylinder radius to tune the resonant frequency across the band gap. The TM mode is found to be highly localized at the defect in the photonic lattice. PMID- 11736124 TI - Stability of attractive Bose-Einstein condensates in a periodic potential. AB - Using a standing light wave potential, a stable quasi-one-dimensional attractive dilute-gas Bose-Einstein condensate can be realized. In a mean-field approximation, this phenomenon is modeled by the cubic nonlinear Schrodinger equation with attractive nonlinearity and an elliptic function potential of which a standing light wave is a special case. New families of stationary solutions are presented. Some of these solutions have neither an analog in the linear Schrodinger equation nor in the integrable nonlinear Schrodinger equation. Their stability is examined using analytic and numerical methods. Trivial-phase solutions are experimentally stable provided they have nodes and their density is localized in the troughs of the potential. Stable time-periodic solutions are also examined. PMID- 11736125 TI - Complexity and regularity of vector-soliton collisions. AB - In this paper, we extensively investigate the collision of vector solitons in the coupled nonlinear Schrodinger equations. First, we show that for collisions of orthogonally polarized and equal-amplitude vector solitons, when the cross-phase modulational coefficient beta is small, a sequence of reflection windows similar to that in the phi(4) model arises. When beta increases, a fractal structure unlike phi(4)'s gradually emerges. But when beta is greater than one, this fractal structure disappears. Analytically, we explain these collision behaviors by a variational model that qualitatively reproduces the main features of these collisions. This variational model helps to establish that these window sequences and fractal structures are caused entirely or partially by a resonance mechanism between the translational motion and width oscillations of vector solitons. Next, we investigate collision dependence on initial polarizations of vector solitons. We discovered a sequence of reflection windows that is phase induced rather than resonance induced. Analytically, we have derived a simple formula for the locations of these phase-induced windows, and this formula agrees well with the numerical data. Last, we discuss collision dependence on relative amplitudes of initial vector solitons. We show that when vector solitons have different amplitudes, the collision structure simplifies. Feasibility of experimental observation of these results is also discussed at the end of the paper. PMID- 11736126 TI - Adiabatic interaction of N ultrashort solitons: universality of the complex Toda chain model. AB - Using the Karpman-Solov'ev method we derive the equations for the two-soliton adiabatic interaction for solitons of the modified nonlinear Schrodinger equation (MNSE). Then we generalize these equations to the case of N interacting solitons with almost equal velocities and widths. On the basis of this result we prove that the N MNSE-soliton train interaction (N>2) can be modeled by the completely integrable complex Toda chain (CTC). This is an argument in favor of universality of the complex Toda chain that was previously shown to model the soliton train interaction for nonlinear Schrodinger solitons. The integrability of the CTC is used to describe all possible dynamical regimes of the N-soliton trains that include asymptotically free propagation of all N solitons, N-soliton bound states, various mixed regimes, etc. It allows also to describe analytically the manifolds in the 4N-dimensional space of initial soliton parameters that are responsible for each of the regimes mentioned above. We compare the results of the CTC model with the numerical solutions of the MNSE for two and three-soliton interactions and find a very good agreement. PMID- 11736127 TI - Perturbation theory for domain walls in the parametric Ginzburg-Landau equation. AB - We demonstrate that in the parametrically driven Ginzburg-Landau equation arbitrarily small nongradient corrections lead to qualitative differences in the dynamical properties of domain walls in the vicinity of the transition from rest to motion. These differences originate from singular rotation of the eigenvector governing the transition. We present analytical results on the stability of Ising walls, deriving explicit expressions for the critical eigenvalue responsible for the transition from rest to motion. We then develop a weakly nonlinear theory to characterize the singular character of the transition and analyze the dynamical effects of spatial inhomogeneities. PMID- 11736128 TI - Nonlinear modulation of multidimensional lattice waves. AB - The equations governing weakly nonlinear modulations of N-dimensional lattices are considered using a quasidiscrete multiple-scale approach. It is found that the evolution of a short wave packet for a lattice system with cubic and quartic interatomic potentials is governed by the generalized Davey-Stewartson (GDS) equations, which include mean motion induced by the oscillatory wave packet through cubic interatomic interaction. The GDS equations derived here are more general than those known in the theory of water waves because of the anisotropy inherent in lattices. The generalized Kadomtsev-Petviashvili equations describing the evolution of long-wavelength acoustic modes in two- and three-dimensional lattices are also presented. Then the modulational instability of an N dimensional Stokes lattice wave is discussed based on the N-dimensional GDS equations obtained. Finally, the one- and two-soliton solutions of two dimensional GDS equations are provided by means of Hirota's bilinear transformation method. PMID- 11736129 TI - Propagation of wave packets in randomly stratified media. AB - The propagation of a narrow-band signal radiated by a point source in a randomly layered absorbing medium is studied asymptotically in the weak-scattering limit. It is shown that in a disordered stratified medium that is homogeneous on average, a pulse is channelled along the layers in a narrow strip in the vicinity of the source. The space-time distribution of the pulse energy is calculated. Far from the source, the shape of wave packets is universal and independent of the frequency spectrum of the radiated signal. Strong localization effects manifest themselves also as a low-decaying tail of the pulse and a strong time delay in the direction of stratification. The frequency-momentum correlation function in a one-dimensional random medium is calculated. PMID- 11736130 TI - Rotating annulus experiment: large-scale helical soliton in the atmosphere? AB - A typhoon is a cyclone vortex with a warm low pressure center, formed over tropical oceanic waters. A large-scale rotating annulus experiment of fluid dynamics is carried out, under the conditions of dynamic similarity, geometric similarity, and the similarity of boundary conditions. In the first step, with the help of infrared heaters, the basic flow field and helical structure of a single typhoon were successfully simulated; then two model typhoons were generated, and their interactions tested. It demonstrated that they did separate after colliding with each other, and their respective basic shapes were restored, which confirms the basic dynamic features of typhoons in nature as solitons. It was also shown that the formation of their helical structures is related to the adapting process of atmosphere to the rotation of the earth and that their dynamic characteristics as solitons come from a result of an equilibrium between their dispersion and the nonlinear convergence of the anticyclones, with whose combined actions their structure remains stable for a long period, which in turn means that they are indeed three-dimensional helical solitons. PMID- 11736131 TI - Asynchronous development of the Benjamin-Feir unstable mode: solution of the Davey-Stewartson equation. AB - The long time evolution of the Benjamin-Feir unstable mode in two dimension is described by the growing-and-decaying mode solution to the Davey-Stewartson equation. The solution of the hyperbolic Davey-Stewartson (the so-called Davey Stewartson I) equation is analyzed to show that the resonance between line soliton and growing-and-decaying mode exists. If the resonant condition is exactly satisfied, the growing-and-decaying mode exists only in the forward region of propagation of soliton and the soliton is accelerated (or decelerated). Under the quasiresonant condition, the growing-and-decaying mode grows at first in the forward region, and after the sequence of the evolution has done in the forward region the mode starts to grow in the backward region of the soliton. PMID- 11736132 TI - Resolvent method for computations of localized defect modes of H-polarization in two-dimensional photonic crystals. AB - We have developed and tested a version of the resolvent (or Green's function) method, based on the shift-inverse of the Maxwellian operator, that ensures stable convergence of iterative computations of localized defect modes of H polarization in two-dimensional (2D) photonic crystals. The defect states are obtained by solving the eigenvalue problem for an associated compact operator with the expansion in Bloch eigenfunctions of the unperturbed Maxwellian operator. This method can be extended to 3D photonic crystals. We apply the method to a 2D square lattice of square dielectric rods in a dielectric background and compute (with controlled precision of approximately 1%) the defect modes induced by the replacement of one rod (the defect). We investigate the rise and variation of the defect frequencies in a photonic band gap, caused by the increase of the dielectric strength of the defect, for four branches of localized modes of various symmetries. PMID- 11736133 TI - Heterogeneous versus discrete mapping problem. AB - We propose a method for mapping a spatially discrete problem, stemming from the spatial discretization of a parabolic or hyperbolic partial differential equation of gradient type, to a heterogeneous one with certain comparable dynamical features pertaining, in particular, to coherent structures. We focus the analysis on a (1+1)-dimensional phi(4) model and confirm the theoretical predictions numerically. We also discuss possible generalizations of the method and the ensuing qualitative analogies between heterogeneous and discrete systems and their dynamics. PMID- 11736134 TI - Wave propagation in media having negative permittivity and permeability. AB - Wave propagation in a double negative (DNG) medium, i.e., a medium having negative permittivity and negative permeability, is studied both analytically and numerically. The choices of the square root that leads to the index of refraction and the wave impedance in a DNG medium are determined by imposing analyticity in the complex frequency domain, and the corresponding wave properties associated with each choice are presented. These monochromatic concepts are then tested critically via a one-dimensional finite difference time domain (FDTD) simulation of the propagation of a causal, pulsed plane wave in a matched, lossy Drude model DNG medium. The causal responses of different spectral regimes of the medium with positive or negative refractive indices are studied by varying the carrier frequency of narrowband pulse excitations. The smooth transition of the phenomena associated with a DNG medium from its early-time nondispersive behavior to its late-time monochromatic response is explored with wideband pulse excitations. These FDTD results show conclusively that the square root choice leading to a negative index of refraction and positive wave impedance is the correct one, and that this choice is consistent with the overall causality of the response. An analytical, exact frequency domain solution to the scattering of a wave from a DNG slab is also given and is used to characterize several physical effects. This solution is independent of the choice of the square roots for the index of refraction and the wave impedance, and thus avoids any controversy that may arise in connection with the signs of these constituents. The DNG slab solution is used to critically examine the perfect lens concept suggested recently by Pendry. It is shown that the perfect lens effect exists only under the special case of a DNG medium with epsilon(omega)=mu(omega)=-1 that is both lossless and nondispersive. Otherwise, the closed form solutions for the field structure reveal that the DNG slab converts an incident spherical wave into a localized beam field whose parameters depend on the values of epsilon and mu. This beam field is characterized with a paraxial approximation of the exact DNG slab solution. These monochromatic concepts are again explored numerically via a causal two dimensional FDTD simulation of the scattering of a pulsed cylindrical wave by a matched, lossy Drude model DNG slab. These FDTD results demonstrate conclusively that the monochromatic electromagnetic power flow through the DNG slab is channeled into beams rather then being focused and, hence, the Pendry perfect lens effect is not realizable with any realistic metamaterial. PMID- 11736135 TI - Numerically complemented analytic method for solving the time-independent one dimensional Schrodinger equation. AB - A general method of solving the one-dimensional Schrodinger equation is developed. The first step is to construct an exactly solvable reference potential of several smoothly joined Morse-type components, which should be a good approximation to a given potential. The exact solutions for that reference Hamiltonian are then combined with a nonperturbative approach [R. G. Gordon, J. Chem. Phys. 51, 14 (1969)], which enables us to numerically solve the energy eigenvalue problem for the original potential to any desired accuracy. A full description of the analytical procedures is given and examples of both exact and numerical solutions, are presented. PMID- 11736136 TI - Probability tree algorithm for general diffusion processes. AB - Motivated by path-integral numerical solutions of diffusion processes, PATHINT, we present a tree algorithm, PATHTREE, which permits extremely fast accurate computation of probability distributions of a large class of general nonlinear diffusion processes. PMID- 11736137 TI - Rossby vortex simulation on a paraboloidal coordinate system using the lattice Boltzmann method. AB - In this paper, we apply our compressible lattice Boltzmann model to a rotating parabolic coordinate system to simulate Rossby vortices emerging in a layer of shallow water flowing zonally in a rotating paraboloidal vessel. By introducing a scaling factor, nonuniform curvilinear mesh can be mapped to a flat uniform mesh and then normal lattice Boltzmann method works. Since the mass per unit area on the two-dimensional (2D) surface varies with the thickness of the water layer, the 2D flow seems to be "compressible" and our compressible model is applied. Simulation solutions meet with the experimental observations qualitatively. Based on this research, quantitative solutions and many natural phenomena simulations in planetary atmospheres, oceans, and magnetized plasma, such as the famous Jovian Giant Red Spot, the Galactic Spiral-vortex, the Gulf Stream, and the Kuroshio Current, etc., can be expected. PMID- 11736138 TI - Hierarchical approach for computing spin glass ground states. AB - We describe a numerical algorithm for computing spin glass ground states with a high level of reliability. The proposed method uses a population based search and applies optimization on multiple scales. Benchmarks are given leading to estimates of the performance on large lattices. PMID- 11736140 TI - Evaluation of atomic integrals for hybrid Gaussian type and plane-wave basis functions via the McMurchie-Davidson recursion formula. AB - A convenient formalism is developed for the evaluation of atomic integrals composed of a hybrid Gaussian type function and plane-wave (GTF-PW) basis set, based upon the recursion scheme proposed by McMurchie and Davidson [L. E. McMurchie and E. R. Davidson, J. Comput. Phys. 26, 218 (1978)] which was originally for Gaussian type basis functions. We show that revisions of recursion relations in the original article are necessary in order to allow systematic production of overlap, kinetic energy, nuclear attraction, and electron repulsion integrals in compact forms. Involving easy calculation of complex incomplete gamma functions, the recursion relations enable the use of hybrid GTF-PW basis functions with arbitrarily large angular momentum. This basis function can be applied to the first-principle calculation for solids involving localized electron orbitals. PMID- 11736139 TI - Semiclassical quantization by harmonic inversion: comparison of algorithms. AB - Harmonic inversion techniques have been shown to be a powerful tool for the semiclassical quantization and analysis of quantum spectra of both classically integrable and chaotic dynamical systems. Various computational procedures have been proposed for this purpose. Our aim is to find out which method is numerically most efficient. To this end, we summarize and discuss the different techniques and compare their accuracies by way of two example systems. PMID- 11736141 TI - Structure determination of organic molecules from diffraction data by simulated annealing. AB - We study simulated annealing techniques for crystal structure determination from diffraction data. We demonstrate that for this problem the efficiency of simulated annealing can be systematically improved by an iterative simulation protocol. Our approach is tested for the example of 9-(methylamino)-1 H-phenalen 1-one-1, 4-dioxan-2-yl hydroperoxide solvate (C18H19NO5). PMID- 11736142 TI - Time-scale invariance of relaxation processes of density fluctuation in slow neutron scattering in liquid cesium. AB - The realization of the idea of time-scale invariance for relaxation processes in liquids has been performed by the memory functions formalism. The best agreement with experimental data for the dynamic structure factor S(k,omega) of liquid cesium near melting point in the range of wave vectors (0.4 A(-1) < or = k < or = 2.55 A(-1)) is found with the assumption of concurrence of relaxation scales for memory functions of third and fourth orders. Spatial dispersion of the first four points in the spectrum of the statistical parameter of non-Markovity epsilon(i)(k,omega) at i=1,2,3,4 has allowed us to reveal the non-Markov nature of collective excitations in liquid cesium, connected with long-range memory effect. PMID- 11736143 TI - Free energy fluctuations for bimodal directed polymers in 1+1 dimensions. AB - By investigating the covariance between the free energy and the entropy, we studied the free energy fluctuations for bimodal directed polymers at finite temperatures. It is proposed that they have a phase transition from the KPZ behavior for pp(c) for the free energy fluctuations at any finite temperature. PMID- 11736144 TI - Magnetic order in the Ising model with parallel dynamics. AB - It is discussed how the equilibrium properties of the Ising model are described by an Hamiltonian with an antiferromagnetic low temperature behavior if only an heat bath dynamics, with the characteristics of a Probabilistic cellular automaton, is assumed to determine the temporal evolution of the system. PMID- 11736145 TI - Ising model on a small world network. AB - A one-dimensional Ising model is studied, via Monte Carlo simulations, on a small world network, where each site has, apart from couplings to its two nearest neighbors, a certain probability to be linked to one of its farther neighbors. It is demonstrated that even a small fraction of such links enables the system to order at finite temperatures. The critical exponent beta is smaller than the two dimensional value, and seems to be independent of the concentration of the extra links. The dependence of the magnetization and the critical temperature on the concentration of the small world links is also presented. PMID- 11736146 TI - Scaling of random spreading in small world networks. AB - In this study we have carried out computer simulations of random walks on Watts Strogatz-type small world networks and measured the mean number of visited sites and the return probabilities. These quantities were found to obey scaling behavior with intuitively reasoned exponents as long as the probability p of having a long range bond was sufficiently low. PMID- 11736147 TI - Mean-field theory for car accidents. AB - We study analytically the occurrence of car accidents in the Nagel-Schreckenberg traffic model. We obtain exact results for the occurrence of car accidents P(ac) as a function of the car density rho and the degree of stochastic braking p(1) in the case of speed limit v(max)=1. Various quantities are calculated analytically. The nontrivial limit p(1)-->0 is discussed. PMID- 11736148 TI - Heuristic model for the energy spectrum of phase turbulence. AB - We present a heuristic model for the energy spectrum of the one-dimensional phase turbulence in the steady state of the Kuramoto-Sivashinsky equation. Our model contains an energy transfer mechanism from low- to high-wave-vector modes. The energy transfer is written as the sum of local and nonlocal interactions. Our analytical results show good agreement with numerical simulations, particularly for the hump in the energy spectrum, which is mainly due to the local interactions. PMID- 11736149 TI - Femtosecond laser-guided electric discharge in air. AB - The filament due to the self-guided propagation of an infrared femtosecond laser pulse in atmospheric-pressure air is used to trigger and guide an electric discharge. The long low density plasma channel due to the filament is first heated by the Joule effect during an initial transient plasma stage. The heated channel of recombined gas then hydrodynamically expands radially. The onset of a discharge starts when the density depression on axis reaches the threshold discharge value. This model is supported by detailed experimental and numerical analysis. PMID- 11736150 TI - Thermodynamic functions of harmonic Coulomb crystals. AB - Phonon frequency moments and thermodynamic functions (electrostatic and vibrational parts of the free energy, internal energy, and heat capacity) are calculated for bcc and fcc Coulomb crystals in the harmonic approximation with a fractional accuracy < or equivalent to 10(-5). Temperature dependence of thermodynamic functions is fitted by analytical formulas with an accuracy of a few parts in 10(5). The static-lattice (Madelung) part of the free energy is calculated with an accuracy of approximately 10(-12). The Madelung constant and frequency moments of hcp crystals are also computed. PMID- 11736151 TI - Dynamics of localized and nonlocalized optical vortex solitons in cubic-quintic nonlinear media. AB - The nonlinear dynamics of laser beams carrying phase singularity in media with cubic-quintic nonlinearity changing from self-focusing to self-defocusing is examined. A novel kind of stable nonlocalized optical vortices appears in such media as well as localized vortex solitons. Linear stability analysis and numerical simulations show the stability of localized vortices only in the defocusing region. PMID- 11736152 TI - Comment on "Critical behavior of a two-species reaction-diffusion problem". AB - In a recent paper, de Freitas et al. [Phys. Rev. E 61, 6330 (2000)] presented simulational results for the critical exponents of the two-species reaction diffusion system A+B-->2B and B-->A in dimension d=1. In particular, the correlation length exponent was found as nu=2.21(5) in contradiction to the well known relation nu=2/d. In this Comment, the symmetry arguments leading to exact critical exponents for the universality class to which this reaction-diffusion system belongs are concisely reconsidered. PMID- 11736154 TI - Comment on "Simple approach to the creation of a strange nonchaotic attractor in any chaotic system". AB - We address the problem of existence of strange nonchaotic attractors (SNAs) in quasiperiodically forced dynamical systems. Recently, Shuai and Wong [Phys. Rev. E 59, 5338 (1999)] suggested a universal method for constructing a SNA in an arbitrary system possessing chaos. We demonstrate here that, in general, this method fails. For arbitrary systems, it gives a SNA only in a vicinity of transition to chaos. We discuss also a special example, where the method by Shuai and Wong indeed produces a SNA. PMID- 11736156 TI - Comment on "Intermittency in chaotic rotations". AB - Lai et al. [Phys. Rev. E 62, R29 (2000)] claim that the angular velocity of the phase point moving along the chaotic trajectory in a properly chosen projection (the instantaneous frequency) is intermittent. Using the same examples, namely the Rossler and the Lorenz systems, we show the absence of intermittency in the dynamics of the instantaneous frequency. This is confirmed by demonstrating that the phase dynamics exhibits normal diffusion. We argue that the nonintermittent behavior is generic. PMID- 11736158 TI - Comment on "Highly nonlinear, sign-varying shift of hydrogen spectral lines in dense plasmas". AB - Possible inconsistencies between the recent hydrogen H(alpha) spectral line shift measurements and modifications of the theory of these shifts by Escarguel et al. [Phys. Rev. E 62, 2667 (2000)], and earlier measurements in dense plasmas and corresponding calculations are discussed. Some of the claimed differences may likely be due to underestimates of Debye shielding effects and to differences between definitions of line shifts in the case of asymmetric profiles. PMID- 11736159 TI - Reconsideration of intermittent synchronization in coupled chaotic pendula. AB - We reinvestigate the intermittent synchronization phenomenon of coupled chaotic pendula that has recently been a controversy. We propose a simple numerical scheme by which one can easily determine whether the observed synchronization is a numerical artifact of computer analysis or not. By using this scheme, for certain coupling strength regime, we find that the average time taken for synchronization linearly depends on the precision of calculations. According to Longa et al.'s criterion for synchronization, this implies that the observed synchronization is genuine. PMID- 11736160 TI - Quantum decay rates for driven barrier potentials in the strong friction limit. AB - Quantum decay rates for barrier potentials driven by external stochastic and periodic forces in the strong damping regime are studied. Based on the quantum Smoluchowski equation derived recently by Ankerhold, Pechukas, and Grabert [Phys. Rev. Lett. 87, 086802 (2001)] explicit analytical and numerical results are presented for the case of the resonant activation phenomenon in a bistable potential and the escape from a metastable well with oscillating barrier, respectively. The significant impact of quantum fluctuations is revealed. PMID- 11736161 TI - Memory in two-dimensional heap experiments. AB - The measurement of force distributions in sandpiles provides a useful way to test concepts and models of the way forces propagate within noncohesive granular materials. Recent theory [J.-P. Bouchaud, M.E. Cates, and P. Claudin, J. Phys. I 5, 639 (1995); M. E. Cates, J. P. Wittmer, J.-P. Bouchaud, and P. Claudin, Phil. Trans. Roy. Soc. 356, 2535 (1998)] by Bouchaud et al. implies that the internal structure of a heap (and therefore the force pathway) is a strong function of the construction history. In general, it is difficult to obtain information that could test this idea from three-dimensional granular experiments except at boundaries. However, two-dimensional systems, such as those used here, can yield information on forces and particle arrangements in the interior of a sample. We obtain position and force information through the use of photoelastic particles. These experiments show that the history of the heap formation has a dramatic effect on the arrangement of particles (texture) and a weaker but clear effect on the forces within the sample. Specifically, heaps prepared by pouring from a point source show strong anisotropy in the contact angle distribution. Depending on additional details, they show a stress dip near the center. Heaps formed from a broad source show relatively little contact angle anisotropy and no indication of a stress dip. PMID- 11736162 TI - Rheology of a confined granular material. AB - We study the rheology of a granular material slowly driven in a confined geometry. The motion is characterized by a steady sliding with a resistance force increasing with the driving velocity and the surrounding relative humidity. For lower driving velocities a transition to stick-slip motion occurs, exhibiting a blocking enhancement with decreasing velocity. We propose a model to explain this behavior pointing out the leading role of friction properties between the grains and the container's boundary. PMID- 11736163 TI - Ferrohydrodynamics: testing a third magnetization equation. AB - A new magnetization equation recently derived from irreversible thermodynamics is employed to the calculation of an increase of ferrofluid viscosity in a magnetic field. Results of the calculations are compared with those obtained on the basis of two well-known magnetization equations. One of the two was obtained phenomenologically, another one was derived microscopically from the Fokker Planck equation. It is shown that the third magnetization equation yields a quite satisfactory description of magnetiviscosity in the entire region of magnetic field strength and the flow vorticity. This equation turns out to be valid-like the microscopically derived equation but unlike the former phenomenological equation-even far from equilibrium, and so it should be recommended for further applications. PMID- 11736164 TI - Critical behavior of nonlinear permittivity in the smectic-A phase of chiral liquid crystals. AB - We study the nonlinear permittivity of chiral liquid crystals in the smectic-A (Sm-A) phase near the ferroelectric smectic-C* (Sm-C*) phase and the smectic C(*)(alpha) (Sm-C(*)(alpha)) phase theoretically and experimentally. The third order nonlinear permittivity epsilon(3) shows the critical behavior with the exponent of four near the Sm-C* phase and its sign depends on the order of the phase transition. In the case of the Sm-A-Sm-C(*)(alpha) phase transition, the sign inversion of epsilon(3), presumably due to the large fluctuation of order parameter, is observed near the transition temperature. PMID- 11736165 TI - Anomalous diffusion and phase relaxation. AB - The diffusion and relaxation of a phase are investigated on the basis of several stochastic models. A simple relation between the diffusional behavior of the extended phase and the relaxation of periodic phase observables is found in the case of Gaussian and Levy distributed increments. In these cases, an anomalous diffusion gives rise to a stretched exponential relaxation of phase observables. Continuous time random walks may lead, even in the case of normal diffusion, to a slow algebraic relaxation. PMID- 11736166 TI - Evaluation of terahertz density of vibrational states from specific-heat data: application to silica glass. AB - The efficiency of the Tikhonov regulation method for the specific-heat phonon spectrum inversion problem is demonstrated. The regulation method is applied to the evaluation of the density of vibrational states from the low-temperature specific heat of a silica glass. The density of states obtained is found to be in good agreement with the results of inelastic neutron scattering. The extracted density of states is used for the analysis of the Raman light-vibration coupling coefficient. It is established that the coupling coefficient increases with frequency faster than approximately nu at nu<10 cm(-1). In the frequency range above 10 cm(-1) the coupling coefficient is found to be in agreement with the results of previous investigations. PMID- 11736167 TI - Dynamics of the spin-boson Hamiltonian by the projection operator technique: applications to electron transfer reactions. AB - A perturbative treatment developed by using the projection operator technique is provided to find dynamics of the spin-boson Hamiltonian in the second order approximation of the subsystem-bath interaction. In the framework of the generalized master equations and in the Markovian approximation it leads to the Redfield/Bloch-type equations. The treatment can be applied to both fast and slow bath cases; in this paper we consider the fast bath case. The relaxation times, energy splitting, and bath-induced renormalization effect of the coherence frequency are discussed and applied to a Lorentzian-Ohmic fast bath. A good agreement between our results and those obtained by the path-integral formalism is obtained. The treatment is applied to electron transfer reactions to test the possibility of apparition of the electronic coherence in certain photosynthetic reaction centers. PMID- 11736168 TI - Critical anomaly of dielectric permittivity for the temperature and pressure paths on approaching the critical consolute point. AB - The experimental results of isothermal pressure dielectric permittivity epsilon studies in a critical mixture characterized by a negative shift of critical temperature induced by pressure (dT(C)/dP<0) are presented. The critical effect is portrayed by the same relation as in previous epsilon(T) and epsilon(P) studies, with the critical exponent alpha=0.12+/-0.03. The advantage of pressure studies is the negligible influence of the correction-to-scaling term and the low frequency Maxwell-Wagner effect. This conclusion is supported by the distortion sensitive derivative analysis of the experimental data. In contrast to previous epsilon(P) studies, carried out in mixtures with dT(C)/dP>0, the critical effect manifests by the bending-up behavior near the critical point. It is suggested that signs of the critical amplitudes of epsilon(P) and epsilon(T) anomalies may be related to the excess volume V(E) and the excess enthalpy H(E), respectively. PMID- 11736169 TI - Density-functional theory of hard-sphere condensation under gravity. AB - The onset of condensation of hard spheres in a gravitational field is studied using density-functional theory (DFT). We find that the local density approximation yields results identical to those obtained previously using the kinetic theory [Physica A 271, 192, (1999)], and a weighted density-functional theory gives qualitatively similar results, namely, that the temperature at which condensation begins at the bottom scales linearly with the weight, diameter, and number of layers of particles. We find also that the different DFT approaches give quantitatively different results for the density profiles at low temperatures. In particular, the weighted density-functional approach reveals the layering of hard spheres in the solid regime. PMID- 11736170 TI - Kinetic equations for diffusion in the presence of entropic barriers. AB - We use the mesoscopic nonequilibrium thermodynamics theory to derive the general kinetic equation of a system in the presence of potential barriers. The result is applied to a description of the evolution of systems whose dynamics is influenced by entropic barriers. We analyze in detail the case of diffusion in a domain of irregular geometry in which the presence of the boundaries induces an entropy barrier when approaching the exact dynamics by a coarsening of the description. The corresponding kinetic equation, named the Fick-Jacobs equation, is obtained, and its validity is generalized through the formulation of a scaling law for the diffusion coefficient which depends on the shape of the boundaries. The method we propose can be useful to analyze the dynamics of systems at the nanoscale where the presence of entropy barriers is a common feature. PMID- 11736171 TI - Order statistics of the trapping problem. AB - When a large number N of independent diffusing particles are placed upon a site of a d-dimensional Euclidean lattice randomly occupied by a concentration c of traps, what is the mth moment of the time t(j,N) elapsed until the first j are trapped? An exact answer is given in terms of the probability Phi(M)(t) that no particle of an initial set of M=N,N-1,...,N-j particles is trapped by time t. The Rosenstock approximation is used to evaluate Phi(M)(t), and it is found that for a large range of trap concentrations the mth moment of t(j,N) goes as x(-m) and its variance as x(-2), x being ln(2/d)(1-c)ln N. A rigorous asymptotic expression (dominant and two corrective terms) is given for for the one-dimensional lattice. PMID- 11736172 TI - Stochastic effects in a thermochemical system with Newtonian heat exchange. AB - We develop a mesoscopic description of stochastic effects in the Newtonian heat exchange between a diluted gas system and a thermostat. We explicitly study the homogeneous Semenov model involving a thermochemical reaction and neglecting consumption of reactants. The master equation includes a transition rate for the thermal transfer process, which is derived on the basis of the statistics for inelastic collisions between gas particles and walls of the thermostat. The main assumption is that the perturbation of the Maxwellian particle velocity distribution can be neglected. The transition function for the thermal process admits a continuous spectrum of temperature changes, and consequently, the master equation has a complicated integro-differential form. We perform Monte Carlo simulations based on this equation to study the stochastic effects in the Semenov system in the explosive regime. The dispersion of ignition times is calculated as a function of system size. For sufficiently small systems, the probability distribution of temperature displays transient bimodality during the ignition period. The results of the stochastic description are successfully compared with those of direct simulations of microscopic particle dynamics. PMID- 11736173 TI - Path-integral study of positronium decay in xenon. AB - We use path-integral Monte Carlo to study the properties of a quantum particle equilibrated in a classical Lennard-Jones fluid. By choosing 2m(e) for its mass, and potential parameters corresponding to xenon, we are able to model the behavior of thermalized positronium above the xenon critical temperature. We carefully study the local distortion of the fluid in the neighborhood of the quantum particle, and use this information to compute the annihilation rate as a function of density on two isotherms. The results compare favorably with experiment below the critical point density. Contrary to accepted views, we demonstrate that positronium remains in a self-trapped state at over twice the critical point density. PMID- 11736174 TI - Kinetic theory of fluidized binary granular mixtures. AB - Balance laws and constitutive relations for a binary granular mixture with unequal granular temperature are derived. The complete pair distribution function for two colliding spheres was assumed to be the product of Maxwellian velocity distributions for each phase. The constitutive relations together with the transport equations form a model for a binary granular mixture with unequal granular temperature. An analytical expression for viscosity of each phase shows the effect of different masses and granular temperatures of particles PMID- 11736175 TI - Three-dimensional hydrodynamic lattice-gas simulations of binary immiscible and ternary amphiphilic flow through porous media. AB - We report the results of a study of multiphase flow in porous media. A Darcy's law for steady multiphase flow was investigated for both binary and ternary amphiphilic flow. Linear flux-forcing relationships satisfying Onsager reciprocity were shown to be a good approximation of the simulation data. The dependence of the relative permeability coefficients on water saturation was investigated and showed good qualitative agreement with experimental data. Nonsteady-state invasion flows were investigated, with particular interest in the asymptotic residual oil saturation. The addition of surfactant to the invasive fluid was shown to significantly reduce the residual oil saturation. PMID- 11736176 TI - Granular drag on a discrete object: shape effects on jamming. AB - We study the drag force on discrete objects with circular cross section moving slowly through a spherical granular medium. Variations in the geometry of the dragged object change the drag force only by a small fraction relative to shape effects in fluid drag. The drag force depends quadratically on the object's diameter as expected. We do observe, however, a deviation above the expected linear depth dependence, and the magnitude of the deviation is apparently controlled by geometrical factors. PMID- 11736177 TI - Mechanism for granular segregation. AB - A process is described that produces horizontal size segregation in a vertically vibrated layer of granular material. The behavior is a consequence of two distinct phenomena that are unique to excited granular media: vibration, which causes the large particles to rise to the top of the layer, and a vibrating base with a sawtooth surface profile, which can produce stratified flows in opposite directions at different heights within the layer. The result of combining these effects is that large and small particles are horizontally driven in opposite directions. The observations reported here are based on computer simulations of granular models in two and three dimensions. PMID- 11736178 TI - Granular flows through vertical pipes controlled by an electric field. AB - The flow of granular nickel particles moving down vertical pipes from a hopper in the presence of a local, horizontal ac electric field is studied experimentally. The flow is initiated by opening the bottom outlet of the pipe after the pipe is fully filled with particles from the hopper. The mass of particles flowing out of the pipe is measured as a function of time by an electronic balance. The time dependence of the steady-state flow rate Q, under each fixed voltage V, is obtained. Depending on the magnitude of V, two types of flow behaviors are observed. For low V ( or =V(c)), no interface exists and the whole region between the hopper and the electrodes are densely filled; only one constant flow rate Q(A2) is observed. (The precise meaning of Q(A2) and Q(B) are defined in the text.) The steady-state flow rates Q(A2) and Q(B) measured for each V, are plotted as a function of V. The flow rate Q(A2) is a monotonically decreasing function of V, which can be approximately fitted by a power law, with an exponent of -0.8, while Q(B) is found to be voltage independent. These features result from a competition between the blocking effect of the electric-field region and the gravity-driven pushing effect from the hopper outlet. The local electric field is able to retard the downward movement of a dense column existing above it, but is ineffective in doing so when the column above is dilute in density. PMID- 11736179 TI - Intrinsic viscosity and the electrical polarizability of arbitrarily shaped objects. AB - The problem of calculating the electric polarizability tensor alpha(e) of objects of arbitrary shape has been reformulated in terms of path integration and implemented computationally. The method simultaneously yields the electrostatic capacity C and the equilibrium charge density. These functionals of particle shape are important in many materials science applications, including the conductivity and viscosity of filled materials and suspensions. The method has been validated through comparison with exact results (for the sphere, the circular disk, touching spheres, and tori), it has been found that 10(6) trajectories yield an accuracy of about four and three significant figures for C and alpha(e), respectively. The method is fast: For simple objects, 10(6) trajectories require about 1 min on a PC. It is also versatile: Switching from one object to another is easy. Predictions have also been made for regular polygons, polyhedra, and right circular cylinders, since these shapes are important in applications and since numerical calculations of high stated accuracy are available. Finally, the path-integration method has been applied to estimate transport properties of both linear flexible polymers (random walk chains of spheres) and lattice model dendrimer molecules. This requires probing of an ensemble of objects. For linear chains, the distribution function of C and of the trace (alpha(e)), are found to be universal in a size coordinate reduced by the chain radius of gyration. For dendrimers, these distribution functions become increasingly sharp with generation number. It has been found that C and alpha(e) provide important information about the distribution of molecular size and shape and that they are important for estimating the Stokes friction and intrinsic viscosity of macromolecules. PMID- 11736180 TI - Thermophoretic and ponderomotive forces in a linear cluster of particles. AB - We investigate amplification of thermophoretic and ponderomotive forces caused by renormalization of static fields in linear clusters of particles with scale separation. We found analytically the dependence of the forces acting on the particles in a cluster as a function of the number of particles in a cluster and material characteristics of the particles and the surrounding fluid. We analytically determined the velocity of stationary motion of particles, velocity distribution in a surrounding viscous fluid, and the thermophoretic force when the particles remain stationary due to the applied constraint forces. PMID- 11736181 TI - Direct measurements of colloidal friction coefficients. AB - The time-independent hydrodynamic forces operating between two hard spheres were studied as a function of the interparticle separation r. Two colloids were positioned a distance r apart using a dual-beam optical tweezer similar to that described by Meiners and Quake [Phys. Rev. Lett. 82, 2211 (1999)]. Brownian fluctuations in the positions of the two spheres were followed by recording the intensity of forward scattered laser light. The two particles move cooperatively as a result of hydrodynamic forces. The strength of the correlation increased strongly with decreasing separation r. We show that the temporal and spatial correlations in the particle displacements are determined by the distance dependence of the pair friction tensor xi. The distance and geometry dependence of the measured friction tensor is found to be in excellent agreement with the predictions of low-Reynolds number calculations. PMID- 11736182 TI - Estimation of fractal dimension of colloidal gels in the presence of multiple scattering. AB - Colloidal dispersions of fluorinated polymer particles with a refractive index very close to that of water, have been used to investigate the effect of multiple scattering on the estimated fractal dimension of colloidal gels, at high-particle volume fractions. The extent of multiple scattering was varied by using cuvettes of different internal diameters, from 3 to 18 mm. Three gelation systems with different sizes and volume fractions of primary particles have been characterized by static light scattering SLS. The obtained results indicate that multiple scattering affects only the magnitude of the scattered radiation, but not the estimated fractal dimension of the gels. This result confirms the conclusion of the theoretical study reported by Chen et al. [Phys. Rev. B 37, 5232 (1988)]. As a further confirmation, the same gels have been formed in a specially designed cell, with only 0.1 mm thickness (where multiple scattering is negligible) and characterized using small-angle neutron scattering (SANS). It is found that the fractal dimension estimated from SANS measurements, without multiple scattering, is the same as that estimated from SLS measurements, in the presence of substantial multiple scattering. PMID- 11736183 TI - Structure of ferrofluid dynamics. AB - The complete magnetodissipative structure of ferrofluid dynamics is derived from general principles, without reference to the angular momentum of the ferromagnetic grains. The results are independent of most microscopic details, and easily interpret two previous experiments. Both the Debye theory and the effective-field theory by Shliomis are shown to be special cases of the new set of equations. PMID- 11736184 TI - Surface ordering in a concentrated suspension of colloidal particles investigated by x-ray scattering methods. AB - The spatial arrangement of colloids near the free surface of a concentrated suspension of colloidal silica in water is investigated by means of x-ray scattering. The weakly charged particles are found to organize in layers along the surface normal direction. The degree of layering decreases with increasing distance from the surface and three layers are identified from the scattering profile. In the lateral direction, the scattering profile indicates a random spatial arrangement of particles at the surface. Based on the findings, a simple structural model for the near surface arrangement of colloidal particles in this system is proposed. PMID- 11736185 TI - Relaxation of surface charge on rotating dielectric spheres: Implications on dynamic electrorheological effects. AB - We have examined the effect of an oscillatory rotation of a polarized dielectric particle. The rotational motion leads to a redistribution of the polarization charge on the surface of the particle. We show that the time-averaged steady state dipole moment is along the field direction, but its magnitude is reduced by a factor that depends on the angular velocity of rotation. As a result, the rotational motion of the particle reduces the electrorheological effect. We further assume that the relaxation of the polarized charge is arised from a finite conductivity of the particle or host medium. We calculate the relaxation time based on the Maxwell-Wagner theory, suitably generalized to include the rotational motion. Analytic expressions for the reduction factor and the relaxation time are given and their dependence on the angular velocity of rotation will be discussed. PMID- 11736186 TI - Scaling with temperature and concentration of the nonlinear rheology of a soft hexagonal phase. AB - The nonlinear rheology of a soft surfactant hexagonal phase is examined. The system exhibits a shear-melting transition from a two-dimensional polycrystalline texture to a liquid of cylinders aligned along the flow [Ramos et al., Langmuir 16, 5846 (2000)]. This dynamic transition is associated with a discontinuity in the stress-strain curve (flow curve). A detailed study of the temperature and concentration dependence of the flow curves is presented. The nonlinear rheology is found to display a scaling behavior, when temperature or concentration are varied. We demonstrate that the whole behavior of the hexagonal phase under shear is essentially governed by the linear shear modulus of the sample, G0. When temperature is varied, we show that the two key parameters, which control G0 and in turn, the flow curve, are a transition temperature T(c) and an activation energy E(A). We propose E(A) to be related to the scission energy of one cylinder into two pieces. PMID- 11736187 TI - Breakup of dipolar rings under a perpendicular magnetic field. AB - An experimental and theoretical study of the breakup process of rings, formed by magnetic microspheres, under the application of an external magnetic field perpendicular to the plane of the ring is presented. We found experimentally that when the value of the external magnetic field falls below a lower critical field the dipoles rotate in the ring without any distortion of the ring structure. However, exceeding the upper critical field causes sudden breakup of the ring into short chains aligned with the field. Between the lower and upper critical fields the system is in a metastable state, and hence, it is very sensitive to external perturbations. The spiral opening was found experimentally to be the lowest energy transition from the ring to the chain conformation. We worked out an analytic approach and we performed computer simulations, the results of which are in good agreement with experiments. PMID- 11736188 TI - Acoustic and relaxation processes in supercooled orthoterphenyl by optical heterodyne transient grating experiment. AB - The dynamics of the fragile glass-forming orthoterphenyl have been investigated by transient grating experiments with an heterodyne detection technique. We measured the relaxation processes of this glass former over more than six decades in time with an excellent signal-to-noise ratio. Acoustic, structural, and thermal relaxations have been clearly identified in a time-frequency window not covered by previous spectroscopic studies and their characteristic dynamic parameters have been measured as a function of temperature and wave vector. A detailed comparison with the density response function, calculated on the basis of generalized hydrodynamic model, has been worked out. PMID- 11736189 TI - Nonlinear ac response of anisotropic composites. AB - When a suspension consisting of dielectric particles having nonlinear characteristics is subjected to a sinusoidal (ac) field, the electrical response will in general consist of ac fields at frequencies of the higher-order harmonics. These ac responses will also be anisotropic. In this work, a self consistent formalism has been employed to compute the induced dipole moment for suspensions in which the suspended particles have nonlinear characteristics, in an attempt to investigate the anisotropy in the ac response. The results showed that the harmonics of the induced dipole moment and the local electric field are both increased as the anisotropy increases for the longitudinal field case, while the harmonics are decreased as the anisotropy increases for the transverse field case. These results are qualitatively understood with the spectral representation. Thus, by measuring the ac responses both parallel and perpendicular to the uniaxial anisotropic axis of the field-induced structures, it is possible to perform real-time monitoring of the field-induced aggregation process. PMID- 11736190 TI - Investigating the microenvironments of inhomogeneous soft materials with multiple particle tracking. AB - We develop a multiple particle tracking technique for making precise, localized measurements of the mechanical microenvironments of inhomogeneous materials. Using video microscopy, we simultaneously measure the Brownian dynamics of roughly one hundred fluorescent tracer particles embedded in a complex medium and interpret their motions in terms of local viscoelastic response. To help overcome the inherent statistical limitations due to the finite imaging volume and limited imaging times, we develop statistical techniques and analyze the distribution of particle displacements in order to make meaningful comparisons of individual particles and thus characterize the diversity and properties of the microenvironments. The ability to perform many local measurements simultaneously allows more precise measurements even in systems that evolve in time. We show several examples of inhomogeneous materials to demonstrate the flexibility of the technique and learn new details of the mechanics of the microenvironments that small particles explore. This technique extends other microrheological methods to allow simultaneous measurements of large numbers of probe particles, enabling heterogeneous samples to be studied more effectively. PMID- 11736191 TI - Lattice-Boltzmann method for the simulation of transport phenomena in charged colloids. AB - We present a simulation scheme based on the lattice-Boltzmann method to simulate the dynamics of charged colloids in an electrolyte. In our model we describe the electrostatics on the level of a Poisson-Boltzmann equation and the hydrodynamics of the fluid by the linearized Navier-Stokes equations. We verify our simulation scheme by means of a Chapman-Enskog expansion. Our method is applied to the calculation of the reduced sedimentation velocity U/U(0) for a cubic array of charged spheres in an electrolyte. We show that we recover the analytical solution first derived by Booth [F. Booth, J. Chem. Phys. 22, 1956 (1954)] for a weakly charged, isolated sphere in an unbounded electrolyte. The present method makes it possible to go beyond the Booth theory, and we discuss the dependence of the sedimentation velocity on the charge of the spheres. Finally we compare our results to experimental data. PMID- 11736192 TI - X-ray reflectivity study on the surface and bulk glass transition of polystyrene. AB - The surfaces of polystyrene (PS) films decorated with gold nanoclusters were investigated by x-ray reflectivity measurements. The thicknesses of the films are much larger than the radii of gyration of the different PS samples. By annealing the films above the glass transition temperature T(g) an embedding process of the clusters into the polymer is detected which is accompanied by a substantial increase in the cluster layer thickness due to Brownian motion. These processes start at a sufficiently low viscosity and may be regarded as a probe of the glass transition in the near surface region of the PS films. Simultaneously the thermal expansion of the entire film and hence its approximate bulk behavior were monitored. Two samples of different molecular weight do not show a significant difference between the surface and bulk T(g) values. PMID- 11736193 TI - Jamming, hysteresis, and oscillation in scalar models for shear thickening. AB - We investigate shear thickening and jamming within the framework of a family of spatially homogeneous, scalar rheological models. These are based on the "soft glassy rheology" model of Sollich et al. [Phys. Rev. Lett. 78, 2020 (1997)], but with an effective temperature x that is a decreasing function of either the global stress sigma or the local strain l. For appropriate x=x(sigma), it is shown that the flow curves include a region of negative slope, around which the stress exhibits hysteresis under a cyclically varying imposed strain rate (.)gamma.A subclass of these x(sigma) have flow curves that touch the (.)gamma=0 axis for a finite range of stresses; imposing a stress from this range jams the system, in the sense that the strain gamma creeps only logarithmically with time t, gamma(t) approximately ln t. These same systems may produce a finite asymptotic yield stress under an imposed strain, in a manner that depends on the entire stress history of the sample, a phenomenon we refer to as history dependent jamming. In contrast, when x=x(l) the flow curves are always monotonic, but we show that some x(l) generate an oscillatory strain response for a range of steady imposed stresses. Similar spontaneous oscillations are observed in a simplified model with fewer degrees of freedom. We discuss this result in relation to the temporal instabilities observed in rheological experiments and stick-slip behavior found in other contexts, and comment on the possible relationship with "delay differential equations" that are known to produce oscillations and chaos. PMID- 11736194 TI - Sliding drops in the diffuse interface model coupled to hydrodynamics. AB - Using a film thickness evolution equation derived recently combining long-wave approximation and diffuse interface theory [L. M. Pismen and Y. Pomeau, Phys. Rev. E 62, 2480 (2000)] we study one-dimensional surface profiles for a thin film on an inclined plane. We discuss stationary flat film and periodic solutions including their linear stability. Flat sliding drops are identified as universal profiles, whose main properties do not depend on mean film thickness. The flat drops are analyzed in detail, especially how their velocity, advancing and receding dynamic contact angles and plateau thicknesses depend on the inclination of the plane. A study of nonuniversal drops shows the existence of a dynamical wetting transition with hysteresis between droplike solutions and a flat film with small amplitude nonlinear waves. PMID- 11736195 TI - Level set approach to reversible epitaxial growth. AB - We generalize the level set approach to model epitaxial growth to include thermal detachment of atoms from island edges. This means that islands do not always grow and island dissociation can occur. We make no assumptions about a critical nucleus. Excellent quantitative agreement is obtained with kinetic Monte Carlo simulations for island densities and island size distributions in the submonolayer regime. PMID- 11736196 TI - Pseudo-Casimir effect in untwisted chiral nematic liquid crystals. AB - We investigate theoretically the pseudo-Casimir force between parallel plates immersed in a chiral nematic liquid crystal. We focus on small-separation limit where the director configuration between the plates inducing strong homeotropic anchoring is uniform. We find that the force is attractive at separations smaller than the crossover distance and repulsive otherwise, and that it diverges logarithmically at the critical distance where the uniform structure is replaced by a distorted structure. We also analyze modifications to the force introduced by magnetic field and comment on the possible detection of the effect. PMID- 11736197 TI - Effect of crosslinker geometry on equilibrium thermal and mechanical properties of nematic elastomers. AB - We study three monodomain (single-crystal) nematic elastomer materials, all side chain siloxane polymers with the same mesogenic groups but with different types of crosslinking: (i) short flexible siloxane linkage affine to the network backbone, (ii) short flexible aliphatic crosslinks miscible with mesogenic side chain groups, and (iii) long segments of main-chain nematic polymer. Equilibrium physical properties of these three systems are very different, especially the spontaneous thermal expansion and anisotropic stress-strain response along and perpendicular to the uniform nematic director. In the latter case, we examine the soft elastic plateau during the director reorientation. We compare the nematic order-parameter Q(T), provided primarily by the side mesogenic groups and relatively constant between the samples, and the average backbone chain anisotropy r(T)=l( parallel)/l( perpendicular), which is strongly affected by the crosslinking geometry. The experimental data is compared quantitatively with theoretical models of nematic elastomers. PMID- 11736198 TI - Molecular dynamics simulations of hard sphere solidification at constant pressure. AB - Molecular dynamics simulations in the NPT ensemble are used to study the dynamics of crystallization processes in hard sphere systems. The simulation method used permits us to follow the dynamics after a sudden pressure or temperature quench in a one-step process without the need of extra densification methods. During the quench a strong correlation between the system density and the crystalline order parameter Q(6) is found. The growth of fcc order in the system over time is observed in detail and compared to Q(6)(t). We compare results for the equation of state on the metastable fluid branch with previous results from constant volume molecular dynamics simulations. Some results for the crystallization of binary hard sphere mixtures are also presented for a number of different size ratios. PMID- 11736199 TI - Orientational order in gravity dispersed clay colloids: a synchrotron x-ray scattering study of Na fluorohectorite suspensions. AB - Colloidal suspensions of clay particles in aqueous salt solutions make ideal model systems for the study of interactions between plate-shaped particles, due to the ease in tuning their electrostatic repulsion with the concentration of the salt. Numerous gel and sol structures are possible, including nematic liquid crystalline order, although only qualitative identification of the latter in clay colloids has been available so far. We present synchrotron x-ray diffraction from gravity dispersed solutions of Na fluorohectorite, a synthetic swelling clay, over a large NaCl concentration range. Our use of liquid scattering techniques allows us to identify regions in which particles reorient from horizontal to vertical alignments in strata coexisting at different heights within the sample. We identify two distinct gel regions characterized by differences in orientational anisotropy and domain size. Our results provide direct evidence for nematic order, as well as unique structural information regarding particle morphology and alignment within each of the colloid phases. PMID- 11736200 TI - Field-dependent tilt and birefringence of electroclinic liquid crystals: theory and experiment. AB - An unresolved issue in the theory of liquid crystals is the molecular basis of the electroclinic effect in the smectic-A phase. Recent x-ray scattering experiments suggest that, in a class of siloxane-containing liquid crystals, an electric field changes a state of disordered molecular tilt in random directions into a state of ordered tilt in one direction. To investigate this issue, we measure the optical tilt and birefringence of these liquid crystals as functions of field and temperature, and we develop a theory for the distribution of molecular orientations under a field. A comparison of theory and experiment confirms that these materials have a disordered distribution of molecular tilt directions that is aligned by an electric field, giving a large electroclinic effect. It also shows that the effective dipole moment, a key parameter in the theory, scales as a power law near the smectic-A--smectic-C transition. PMID- 11736201 TI - Semiflexible polymer in a uniform force field in two dimensions. AB - The conformational properties of a semiflexible polymer chain, anchored at one end in a uniform force field, are studied in a simple two-dimensional model. Recursion relations are derived for the partition function and then iterated numerically. We calculate the angular fluctuations of the polymer about the direction of the force field and the average polymer configuration as functions of the bending rigidity, chain length, chain orientation at the anchoring point, and field strength. PMID- 11736202 TI - Free expansion of elastic filaments. AB - The dynamics of an elastic polymer filament undergoing contour length expansion is studied using computer simulation. The expansion occurs by development of transverse buckling waves that grow through a coarsening process. The growing buckles locally organize into a helical structure with a characteristic persistence length. The helical domain boundaries are eliminated from the relaxing structure by unwinding through the ends of the rod. The growth of the helical domains results in self-propulsive motion of the expanding rod, as one large helix spanning the entire chain relaxes during the late stages of the dynamics. Stability analyses and scaling arguments are provided to explain the simulation results. PMID- 11736203 TI - Photoinduced dynamics in a photosensitive side chain polymeric liquid crystal by quasielastic and inelastic neutron scattering. AB - We report the first study by inelastic (INS) and quasielastic neutron scattering of photoinduced changes in the reorientational and vibrational dynamics of a liquid crystalline side chain polymer. We use the cis-trans photoisomerization transition to take the system out of equilibrium and determine the quasielastic (QE) and inelastic scattering laws on two distinct time windows, corresponding to the time-of-flight (IN6) and backscattering (IN16) spectrometers at Institut Laue Langevin (ILL) (Grenoble). Our investigation was focused on the dynamics of the coupling between the mesogenic side chains and the polymeric main chain, which is connected to the extensive optical writing and memory effects that have been demonstrated in this complex material. We report data on the QE broadening and Debye-Waller factors, as a function of temperature across the glass transition (T(g)=293 K). We also studied the dynamical coupling of side and main chains. We report photoinduced changes on the static structure factor, on the purely elastic scattering fraction, on the low frequency vibrational dynamics (around the boson peak region). In particular, we find that on the space-time scales accessible to the INS techniques there is a time structure in the coupling, and that over longer times and distances the two dynamics are decoupled. PMID- 11736204 TI - Hairpin turn dislocations in two-dimensional smectic phases of long semiflexible polymers. AB - We elucidate hairpin turn dislocations in two-dimensional smectic phases of long semi-flexible polymers. We discuss hairpin shapes, sizes, and free energies. We find that hairpin dislocation core may be, under some circumstances, substantially bigger than the smectic period size. Such hairpin dislocations are accompanied by large voids that are stable equilibrium structures with sizes determined by a competition of the polymer bending elasticity and smectic bulk elasticity. The large size of hairpin voids is associated with a low hairpin energy, much smaller than anticipated before. The actual hairpin shape, size, and energy are all qualitatively sensitive to the detailed nature of smectics. We document this by considering hairpin dislocations in lyotropic smectics (systems stabilized by repulsion between polymers, with a positive osmotic pressure) and in thermotropic smectics (systems stable even at zero osmotic pressure, with a preferred distance between semiflexible molecules). We discuss in detail hairpin dislocations in lyotropic sterically stabilized Smectics as well as in DNA cationic lipid complexes. We elucidate the extinction of hairpin dislocations by annihilations with polymer end points. In lyotropic smectics, rates of these processes are shown to be limited by sluggish reptation of semiflexible molecules, as well as by substantial energy barriers. PMID- 11736205 TI - Retrieval behavior and thermodynamic properties of symmetrically diluted Q-Ising neural networks. AB - The retrieval behavior and thermodynamic properties of symmetrically diluted Q Ising neural networks are derived and studied in replica-symmetric mean-field theory generalizing earlier works on either the fully connected or the symmetrical extremely diluted network. Capacity-gain parameter phase diagrams are obtained for the Q=3, Q=4, and Q=infinity state networks with uniformly distributed patterns of low activity in order to search for the effects of a gradual dilution of the synapses. It is shown that enlarged regions of continuous changeover into a region of optimal performance are obtained for finite stochastic noise and small but finite connectivity. The de Almeida-Thouless lines of stability are obtained for arbitrary connectivity, and the resulting phase diagrams are used to draw conclusions on the behavior of symmetrically diluted networks with other pattern distributions of either high or low activity. PMID- 11736206 TI - Transient structures of wave patterns arising in the wave regeneration of subalpine coniferous forests. AB - In wave-regeneration phenomena observed in the subalpine coniferous forests, mainly consisting of Abies species, the blighted forests present various shapes in the course of development, spots at the initial stage turning into arches and finally into long whitish stripes. Because the wave-regeneration could not be followed in the field without long term studies, a simple model has been elaborated to simulate the various different dieback structures observed in the real forests. This model, based on cellular automata, is employed to analyze the power spectral density of canopy tree height fluctuations in the wave-regenerated forests. The results demonstrate that almost all the dieback structures observed in the field can be generated by this simple model, by varying the wind direction and its strength by some stochasticity. The power spectrum density presents various shapes in the course of development, white noise type at the initial stage turning into Lorentz type and finally into 1/f type power spectrum (spatial Fourier frequency). PMID- 11736207 TI - Collapsing bacterial cylinders. AB - Under special conditions bacteria excrete an attractant and aggregate. The high density regions initially collapse into cylindrical structures, which subsequently destabilize and break up into spherical aggregates. This paper presents a theoretical description of the process, from the structure of the collapsing cylinder to the spacing of the final aggregates. We show that cylindrical collapse involves a delicate balance in which bacterial attraction and diffusion nearly cancel, leading to corrections to the collapse laws expected from dimensional analysis. The instability of a collapsing cylinder is composed of two distinct stages: Initially, slow modulations to the cylinder develop, which correspond to a variation of the collapse time along the cylinder axis. Ultimately, one point on the cylinder pinches off. At this final stage of the instability, a front propagates from the pinch into the remainder of the cylinder. The spacing of the resulting spherical aggregates is determined by the front propagation. PMID- 11736208 TI - Master equation approach to molecular motors. AB - A master equation approach to molecular motors allows us to describe a mechanochemical cyclic system where chemical and translational degrees of freedom are treated on an equal footing. A generalized detailed balance condition in the out-of-equilibrium regime is shown to be compatible with the Fokker-Planck equation in the continuum limit. The Onsager reciprocity relations hold for stationary states close to equilibrium, provided the generalized detailed balance condition is satisfied. Semiphenomenological considerations in the case of motor proteins lead to a discrete kinetics model, for which interesting observable quantities may be directly calculated and compared with experimental data. PMID- 11736209 TI - Universal scaling properties of type-I intermittent chaos in isolated resistance arteries are unaffected by endogenous nitric oxide synthesis. AB - Spontaneous fluctuations in flow in isolated rabbit ear resistance arteries may exhibit almost-periodic behavior interrupted by chaotic bursts that can be classified as type-I Pomeau-Manneville intermittency. This conclusion was supported by the construction of parabolic return maps and identification of the characteristic probability distributions for the number of oscillations per laminar segment (n) associated with the type-I scenario. Pharmacological inhibition of nitric oxide (NO) synthesis by the vascular endothelium modulated the dynamics of the reinjection mechanism, and thus the generic shape of the probability distribution for n. Nevertheless, average laminar length was related to a derived bifurcation parameter epsilon according to power-law scaling of the form approximately epsilon(beta), where the estimated critical exponent beta was close to the theoretical value of -0.5 both in the presence and absence of NO synthesis. PMID- 11736210 TI - Indications of nonlinear deterministic and finite-dimensional structures in time series of brain electrical activity: dependence on recording region and brain state. AB - We compare dynamical properties of brain electrical activity from different recording regions and from different physiological and pathological brain states. Using the nonlinear prediction error and an estimate of an effective correlation dimension in combination with the method of iterative amplitude adjusted surrogate data, we analyze sets of electroencephalographic (EEG) time series: surface EEG recordings from healthy volunteers with eyes closed and eyes open, and intracranial EEG recordings from epilepsy patients during the seizure free interval from within and from outside the seizure generating area as well as intracranial EEG recordings of epileptic seizures. As a preanalysis step an inclusion criterion of weak stationarity was applied. Surface EEG recordings with eyes open were compatible with the surrogates' null hypothesis of a Gaussian linear stochastic process. Strongest indications of nonlinear deterministic dynamics were found for seizure activity. Results of the other sets were found to be inbetween these two extremes. PMID- 11736211 TI - Strategies for the evolution of sex. AB - We find that the hypothesis made by Jan, Stauffer, and Moseley [Theory Biosci. 119, 166 (2000)] for the evolution of sex, namely, a strategy devised to escape extinction due to too many deleterious mutations, is sufficient but not necessary for the successful evolution of a steady state population of sexual individuals within a finite population. Simply allowing for a finite probability for conversion to sex in each generation also gives rise to a stable sexual population, in the presence of an upper limit on the number of deleterious mutations per individual. For large values of this probability, we find a phase transition to an intermittent, multistable regime. On the other hand, in the limit of extremely slow drive, another transition takes place to a different steady state distribution, with fewer deleterious mutations within the population. PMID- 11736212 TI - Removal of DNA-bound proteins by DNA twisting. AB - We present a simple model of how local torsional stress in DNA can eject a DNA bound protein. An estimate of the torque tau(*) required to eject a typical DNA bound protein is made through a two-state model of the equilibrium between the bound and unbound states of the protein. For the familiar case of a nucleosome octamer bound to double-stranded DNA, we find this critical torque to be approximately equal to 9k(B)T. More weakly bound proteins and large (approximately equal to kilobase) loops of DNA are shown to be destabilized by smaller torques of only a few k(B)T. We then use our model to estimate the maximum range R(max) at which a protein can be removed by a transient source of twisting. We model twist strain propagation along DNA by simple dissipative dynamics in order to estimate R(max). Given twist pulses of the type expected to be generated by RNA polymerase and DNA gyrase, we find R(max) approximately equal to 70 and 450 bp, respectively, for critical torques of approximately equal to 2k(B)T. PMID- 11736213 TI - Dynamical behavior of a pacemaker neuron model with fixed delay stimulation. AB - In physiological and pathological conditions, many biological oscillators, such as pacemaker cells, operate under the influence of feedbacks. Fixed delay stimulation is a standard preparation to evaluate the effects of such influences. Through the study of the Hodgkin-Huxley model, we show that such recurrent excitation can lead to regular and irregular discharge trains with interdischarge intervals that are up to several multiples of the period of the oscillator. In other words, we show that recurrent excitation can considerably slow down the firings of the pacemaker. This result contrasts with previous studies of similar preparations that have reported that fixed delay stimulation leads to a bursting pattern in which regimes of high-frequency firing alternate with periods of quiescence. We elucidate the mechanisms underlying the behavior of the oscillator under fixed delay perturbation through the analysis of the dynamics of a well known two-dimensional oscillator, namely, the Poincare oscillator. PMID- 11736214 TI - Entropy and local uncertainty of data from sensory neurons. AB - We present an empirical comparison between neural interspike interval sequences obtained from two different kinds of sensory receptors. Both differ in their internal structure as well as in the strength of correlations and the degree of predictability found in the respective spike trains. As a further tool in this context, we suggest the local uncertainty, assigning a well-defined predictability to individual spikes. The local uncertainty is demonstrated to reveal significant patterns within the interspike interval sequences, even when its overall structure is (almost) random. Our approach is based on the concept of symbolic dynamics and information theory. PMID- 11736215 TI - Cavity approach to noisy learning in nonlinear perceptrons. AB - We analyze the learning of noisy teacher-generated examples by nonlinear and differentiable student perceptrons using the cavity method. The generic activation of an example is a function of the cavity activation of the example, which is its activation in the perceptron that learns without the example. Mean field equations for the macroscopic parameters and the stability condition yield results consistent with the replica method. When a single value of the cavity activation maps to multiple values of the generic activation, there is a competition in learning strategy between preferentially learning an example and sacrificing it in favor of the background adjustment. We find parameter regimes in which examples are learned preferentially or sacrificially, leading to a gap in the activation distribution. Full phase diagrams of this complex system are presented, and the theory predicts the existence of a phase transition from poor to good generalization states in the system. Simulation results confirm the theoretical predictions. PMID- 11736216 TI - Correlation functions for diffusion-limited annihilation, A+A-->0. AB - The full hierarchy of multiple-point correlation functions for diffusion-limited annihilation, A+A-->0, is obtained analytically and explicitly, following the method of intervals. In the long-time asymptotic limit, the correlation functions of annihilation are identical to those of coalescence, A+A-->A, despite differences between the two models in other statistical measures, such as the interparticle distribution function. PMID- 11736217 TI - Coupling of thermal and mass diffusion in regular binary thermal lattice gases. AB - We have constructed a regular binary thermal lattice gas model in which the thermal diffusion and mass diffusion are coupled and form two nonpropagating diffusive modes. The power spectrum is shown to be similar in structure as for the one in real fluids, in which the central peak becomes a combination of coupled entropy and concentration contributions. Our theoretical findings for the power spectra are confirmed by computer simulations performed on this model. PMID- 11736218 TI - Thermodynamics of the asymmetric double sinh-Gordon theory in 1+1 dimensions. AB - Classical thermodynamics of the (1+1)-dimensional asymmetric double sinh-Gordon system is investigated. The pseudo-Schrodinger equation resulting from the transfer integral method is solved numerically and within the semiclassical approximation; the exact results are also given at several temperatures. It is found that the specific heat exhibits a characteristic hump resembling a similar one observed in the systems with a symmetric potential; in some structures, extremely narrow and extremely high peak is developed. The interpretation for this behavior is given. PMID- 11736219 TI - Region visited by a spherical Brownian particle in the presence of an absorbing boundary. AB - We calculate the time dependence of the average volume of a Wiener sausage in the presence of an absorbing boundary in one and three dimensions. In one dimension it is shown that the presence of an absorbing point reduces the time dependence of the average span from being proportional to sqrt[t] in an unbounded space, to being proportional to ln(t) at long times. In three dimensions the average volume increases as sqrt[t] at long times rather than being proportional to t as in free space. PMID- 11736220 TI - Classical density functional theory of freezing in simple fluids: numerically induced false solutions. AB - Density functional theory (DFT) has provided many insights into the freezing of simple fluids. Several analytical and numerical solution have shown that the DFT provides an accurate description of freezing of hard spheres and their mixtures. Compared to other techniques, numerical, grid-based algorithms for solving the DFT equations have more variational freedom and are capable of describing subtle behavior, as that seen in mixtures with multipeaked density profiles. However the grid-based approach is sensitive to the coarseness of the mesh employed. Here we summarize how the granularity of the mesh affects the freezing point within the DFT. For coarse meshes, we show that the freezing point is masked by numerically induced false minima of the DFT grand potential. These false minima are removed when a fine enough grid is used to represent properly the density profiles. Our results suggest that others using such grid-based methods have focused on such numerical artifacts that have little to do with real phenomena. PMID- 11736221 TI - Temperature dependence of the coefficient of electronic polarizability in calamitic nematic liquid crystals. AB - In this report the temperature dependence of the coefficient of electronic polarizability (straight phi(i)) is determined by means of the thermal expansion coefficient (beta) and ordinary/extraordinary refractive indexes measurements near the calamitic nematic-isotropic phase transition in a lyotropic mixture of sodium decylsulphonate, decanol, and water. These parameters (straight phi(i) and beta) were related to the extraordinary and ordinary refractive indexes via the temperature derivative of the Vuks's equation. The results showed that near the nematic-isotropic phase transition, the measured value of straight phi(i) was found to be about two orders of magnitude smaller than that obtained for thermotropic, showing a remarkable difference in the molecular electronic polarizability strength between lyotropic and thermotropic liquid crystals. PMID- 11736222 TI - Comment on "Magnetoviscosity and relaxation in ferrofluids". AB - It is shown and discussed how the conventional system of hydrodynamic equations for ferrofluids was derived. The set consists of the equation of fluid motion, the Maxwell equations, and the magnetization equation. The latter was recently revised by Felderhof [Phys. Rev. E 62, 3848 (2000)]. His phenomenological magnetization equation looks rather like our corresponding equation, but leads to wrong consequences for the dependence of ferrofluid viscosity and magnetization relaxation time on magnetic field. PMID- 11736224 TI - Comment on "Spin-1 aggregation model in one dimension". AB - Girardi and Figueiredo have proposed a simple model of aggregation in one dimension to mimic the self assembly of amphiphiles in aqueous solution [Phys. Rev. E 62, 8344 (2000)]. We point out that interesting results can be obtained if a different set of interactions is considered, instead of their choice (the s=1 Ising model). PMID- 11736225 TI - Transmission fluctuations in chaotic microwave billiards with and without time reversal symmetry. AB - Transmission fluctuations have been studied in a microwave billiard in dependence to the number of attached wave guides on its entrance and exit. To investigate the influence of breaking time-reversal symmetry, ferrite cylinders were introduced into the billiard. The obtained transmission intensity distributions are compared with predictions from the random matrix theory. Because of the strong absorption caused by the ferrites, the existing statistical scattering theories had to be modified, by incorporating a number of additional absorbing scattering channels. PMID- 11736226 TI - Failure of random matrix theory to correctly describe quantum dynamics. AB - Consider a classically chaotic system that is described by a Hamiltonian H(0). At t=0 the Hamiltonian undergoes a sudden change (H)0-->H. We consider the quantum mechanical spreading of the evolving energy distribution, and argue that it cannot be analyzed using a conventional random-matrix theory (RMT) approach. Conventional RMT can be trusted only to the extent that it gives trivial results that are implied by first-order perturbation theory. Nonperturbative effects are sensitive to the underlying classical dynamics, and therefore the Planck's over 2 pi-->0 behavior for effective RMT models is strikingly different from the correct semiclassical limit. PMID- 11736227 TI - Onset of wave fronts in a discrete bistable medium. AB - The transition from a standing front to a traveling front is studied in an array of symmetric bistable coupled oscillators. The mechanism leading to propagation may be understood in the context of a gluing bifurcation involving a pair of homoclinic loops. The velocity of the front shows a logarithmic dependence with the coupling strength according to this mechanism. PMID- 11736228 TI - Scaling of the velocity power spectra in turbulent thermal convection. AB - We report measurements of the local velocity in a convection cell filled with water. In the buoyancy subrange (the scales between the Bolgiano scale f(B) and the integral scale f(o)), two scaling regions of the velocity frequency power spectra, separated by the peak frequency f(p) of the dissipation spectra, are found. For f(p) and the two-time autocorrelations . We also compute the persistence properties of a single spin, of local magnetization, and of domains. The analogous quantities for the +/-J Ising spin glass in an applied field are obtained from the RFIM via a gauge transformation. In addition to these we compute the two-point two-time correlation function which can in principle be measured by experiments on spin-glass-like systems. The thermal fluctuations are studied and found to be dominated by rare events; in particular all moments of truncated equal time correlations are computed. Physical properties which are typically measured in aging experiments are also studied, focusing on the response to a small magnetic field which is applied after waiting for the system to equilibrate for a time t(w). The nonequilibrium fluctuation-dissipation ratio X(t,t(w)) is computed. We find that for (t-t(w)) approximately t(alpha)(w) with alpha;<1, the ratio equal to its equilibrium value X=1, although time translational invariance does not hold in this regime. For t-t(w) approximately t(w) the ratio exhibits an aging regime with a nontrivial X=X(t/t(w)) not equal to 1, but the behavior is markedly different from mean field theory. Finally the distribution of the total magnetization and of the number of domains is computed for large finite size systems. General issues about convergence toward equilibrium and the possibilities of weakly history-dependent evolution in other random systems are discussed. PMID- 11736238 TI - Interface depinning in the absence of an external driving force. AB - We study the pinning-depinning phase transition of interfaces in the quenched Kardar-Parisi-Zhang model as the external driving force F goes towards zero. For a fixed value of the driving force, we induce depinning by increasing the nonlinear term coefficient lambda, which is related to lateral growth, up to a critical threshold. We focus on the case in which there is no external force applied (F=0) and find that, contrary to a simple scaling prediction, there is a finite value of lambda that makes the interface to become depinned. The critical exponents at the transition are consistent with directed percolation depinning. PMID- 11736237 TI - Thermodynamics and diffusion of a lattice gas on a simple cubic lattice. AB - A lattice gas model with nearest neighbor attractive interactions on a simple cubic lattice is considered. The method of nonequilibrium statistical ensembles due to Zubarev is used to derive expressions for jump and chemical diffusion coefficients. For thermally activated hopping dynamics in the hydrodynamical (low frequency, long wavelength) limit, and neglecting specific memory effects, these expressions are represented in a simple form in terms of the zero concentration limit of the chemical diffusion coefficient and equilibrium characteristics, i.e., the chemical potential, and the probability for two nearest neighbor sites to be vacant. These equilibrium characteristics are calculated by means of the self-consistent diagram approximation. The equilibrium characteristics and diffusion coefficients are in a good agreement with extensive Monte Carlo simulation results. PMID- 11736239 TI - Anomalous percolation properties of growing networks. AB - We describe the anomalous phase transition of the emergence of the giant connected component in scale-free networks growing under mechanism of preferential linking. We obtain exact results for the size of the giant connected component and the distribution of vertices among connected components. We show that all the derivatives of the giant connected component size S over the rate b of the emergence of new edges are zero at the percolation threshold b(c), and S infinity exp[-d(gamma)(b-b(c))(-1/2)], where the coefficient d is a function of the degree distribution exponent gamma. In the entire phase without the giant component, these networks are in a "critical state." The probability P(k) that a vertex belongs to a connected component of a size k is of a power-law form. At the phase transition point, P(k) approximately 1/(k ln k)(2). In the phase with the giant component, P(k) has an exponential cutoff at k(c) approximately 1/S. In the simplest particular case, we present exact results for growing exponential networks. PMID- 11736240 TI - Random sequential adsorption of binary mixtures on a line. AB - A jamming coverage for the random sequential adsorption of binary mixtures of segments on the infinite line is derived. It always appears to be smaller than the coverage associated with the car parking problem. This has to be contrasted with dicrete models, where the coverage of the lattice by mixtures of segments of different sizes is more efficient than by single species. PMID- 11736241 TI - Infection dynamics on scale-free networks. AB - We discuss properties of infection processes on scale-free networks, relating them to the node-connectivity distribution that characterizes the network. Considering the epidemiologically important case of a disease that confers permanent immunity upon recovery, we derive analytic expressions for the final size of an epidemic in an infinite closed population and for the dependence of infection probability on an individual's degree of connectivity within the population. As in an earlier study [R. Pastor-Satorras and A. Vesipignani, Phys. Rev. Lett. 86, 3200 (2001); Phys. Rev. E. 63, 006117 (2001)] for an infection that did not confer immunity upon recovery, the epidemic process--in contrast with many traditional epidemiological models--does not exhibit threshold behavior, and we demonstrate that this is a consequence of the extreme heterogeneity in the connectivity distribution of a scale-free network. Finally, we discuss effects that arise from finite population sizes, showing that networks of finite size do exhibit threshold effects: infections cannot spread for arbitrarily low transmission probabilities. PMID- 11736242 TI - Monte Carlo simulation of O(2) phi(4) field theory in three dimensions. AB - Using standard numerical Monte Carlo lattice methods, we study non-universal properties of the phase transition of three-dimensional straight phi(4) theory of a two-component real field phi=(phi(1),phi(2)) with O(2) symmetry. Specifically, we extract the renormalized values of /u and r/u(2) at the phase transition, where the continuum action of the theory is integral d(3)x[1/2/inverted Delta phi/(2) + 1 2r phi(2)+(u/4!)phi(4)]. These values have applications to calculating the phase-transition temperature of dilute or weakly interacting Bose gases (both relativistic and nonrelativistic). In passing, we also provide perturbative calculations of various O(a) lattice-spacing errors in three-dimensional O(N) scalar field theory, where a is the lattice spacing. PMID- 11736243 TI - Self-consistent generalized Langevin equation for colloid dynamics. AB - We present a general self-consistent theory of colloid dynamics which, for a system without hydrodynamic interactions, allows us to calculate F(k,t), and its self-diffusion counterpart F(S)(k,t), given the effective interaction pair potential u(r) between colloidal particles, and the corresponding equilibrium static structural properties. This theory is build upon the exact results for F(k,t) and F(S)(k,t) in terms of a hierarchy of memory functions, derived from the application of the generalized Langevin equation formalism, plus the proposal of Vineyard-like connections between F(k,t) and F(S)(k,t) through their respective memory functions, and a closure relation between these memory functions and the time-dependent friction function Delta zeta(t). As an illustrative application, we present and analyze a selection of numerical results of this theory in the short- and intermediate-time regimes, as applied to a two dimensional repulsive Yukawa Brownian fluid. For this system, we find that our theory accurately describes the dynamic properties contained in F(k,t) in a wide range of conditions, including strongly correlated systems, at the longest times available from our computer simulations. PMID- 11736244 TI - Enhanced sampling in numerical path integration: an approximation for the quantum statistical density matrix based on the nonextensive thermostatistics. AB - Here we examine a proposed approximation for the quantum statistical density matrix motivated by the nonextensive thermostatistics of Tsallis and co-workers. The approximation involves replacing the physical potential energy with an effective one, corresponding to a generalized nonextensive statistical ensemble. We examine the convergence properties of averages calculated using the effective potential, and introduce a related method for enhanced sampling in numerical path integration. As a necessary measure, path integral energy estimators are introduced for potentials that involve explicit temperature dependence. This sampling method is found to be effective for path integral simulations involving broken ergodicity. PMID- 11736245 TI - Complex-temperature phase diagrams for the q-state Potts model on self-dual families of graphs and the nature of the q-->infinity limit. AB - Exact calculations of the Potts model partition function Z(G,q,v) have been presented for arbitrary q and temperature-like variable v on self-dual strip graphs G of the square lattice with fixed width L(y) and arbitrarily great length Lx with two types of boundary conditions. Letting Lx-->infinity, the resultant free energy and complex-temperature phase diagram have been computed, including the locus B where the free energy is nonanalytic. Results are analyzed for widths L(y)=1,2,3. These results have been used to study the approach to the large-q limit of B. PMID- 11736246 TI - Rigidity of random networks of stiff fibers in the low-density limit. AB - Rigidity percolation is analyzed in two-dimensional random networks of stiff fibers. As fibers are randomly added to the system there exists a density threshold q=q(min) above which a rigid stress-bearing percolation cluster appears. This threshold is found to be above the connectivity percolation threshold q=q(c) such that q(min)=(1.1698+/-0.0004)q(c). The transition is found to be continuous, and in the universality class of the two-dimensional central force rigidity percolation on lattices. At percolation threshold the rigid backbone of the percolating cluster was found to break into rigid clusters, whose number diverges in the limit of infinite system size, when a critical bond is removed. The scaling with system size of the average size of these clusters was found to give a new scaling exponent delta=1.61+/-0.04. PMID- 11736247 TI - Quantum effects after decoherence in a quenched phase transition. AB - We study a quantum mechanical toy model that mimics some features of a quenched phase transition. Both by virtue of a time-dependent Hamiltonian or by changing the temperature of the bath we are able to show that even after classicalization has been reached, the system may display quantum behavior again. We explain this behavior in terms of simple nonlinear analysis and estimate relevant time scales that match the results of numerical simulations of the master equation. This opens new possibilities both in the study of quantum effects in nonequilibrium phase transitions and in general time-dependent problems where quantum effects may be relevant even after decoherence has been completed. PMID- 11736248 TI - Headways in traffic flow: remarks from a physical perspective. AB - Traffic flow can be understood as a realization of a broad class of one dimensional physical systems, where a hard core repulsive interaction competes with a longer ranged attraction between the particles. It can be shown rigorously that the statistical properties of such systems in thermal equilibrium are well described by a family of distributions that stems from the random matrix theory. Analyzing the traffic data from different sources, we show that traffic on real roads belongs to that class of random matrix distributions. Also, various traffic simulation models show a similar behavior. It is demonstrated in such a way that the headway distribution of a highway traffic, that serves usually as a paradigm of systems driven far from equilibrium, is reasonably well described by a distribution originating from equilibrium statistical physics. PMID- 11736249 TI - One-dimensional quantum models with correlated disorder versus classical oscillators with colored noise. AB - We perform an analytical study of the correspondence between a classical oscillator with frequency perturbed by a colored noise and the one-dimensional Anderson-type model with weak correlated diagonal disorder. It is rigorously shown that localization of electronic states in the quantum model corresponds to exponential divergence of nearby trajectories of the classical random oscillator. We discuss the relation between the localization length for the quantum model and the rate of energy growth for the stochastic oscillator. Finally, we examine the problem of electron transmission through a finite disordered lattice by considering the evolution of the classical oscillator. PMID- 11736250 TI - Spectral properties of a generalized chiral Gaussian unitary ensemble. AB - We consider a generalized chiral Gaussian Unitary ensemble (chGUE) based on a weak confining potential. We study the spectral correlations close to the origin in the thermodynamic limit. We show that for eigenvalues separated up to the mean level spacing, the spectral correlations coincide with those of chGUE. Beyond this point, the spectrum is described by an oscillating number variance centered around a constant value. We argue that the origin of such a rigid spectrum is due to the breakdown of the translational invariance of the spectral kernel in the bulk of the spectrum. Finally, we compare our results with the ones obtained from a critical chGUE recently reported in the literature. We conclude that our generalized chGUE does not belong to the same class of universality as the above mentioned model. PMID- 11736251 TI - Bursts in a fiber bundle model with continuous damage. AB - We study the constitutive behavior, the damage process, and the properties of bursts in the continuous damage fiber bundle model introduced recently. Depending on its two parameters, the model provides various types of constitutive behaviors including macroscopic plasticity. Analytical results are obtained to characterize the damage process along the plastic plateau under strain controlled loading; furthermore, for stress controlled experiments we develop a simulation technique, and numerically explore the distribution of bursts of fiber breaks assuming an infinite range of interaction. Simulations revealed that under certain conditions power law distribution of bursts arises with an exponent significantly different from the mean field exponent 5/2. A phase diagram of the model characterizing the possible burst distributions is constructed. PMID- 11736252 TI - Generalized empty-interval method applied to a class of one-dimensional stochastic models. AB - In this work we study, on a finite and periodic lattice, a class of one dimensional (bimolecular and single-species) reaction-diffusion models that cannot be mapped onto free-fermion models. We extend the conventional empty interval method, also called interparticle distribution function (IPDF) method, by introducing a string function, which is simply related to relevant physical quantities. As an illustration, we specifically consider a model that cannot be solved directly by the conventional IPDF method and that can be viewed as a generalization of the voter model and/or as an epidemic model. We also consider the reversible diffusion-coagulation model with input of particles and determine other reaction-diffusion models that can be mapped onto the latter via suitable similarity transformations. Finally we study the problem of the propagation of a wave front from an inhomogeneous initial configuration and note that the mean field scenario predicted by Fisher's equation is not valid for the one dimensional (microscopic) models under consideration. PMID- 11736253 TI - Quantum thermodynamics of systems with anomalous dissipative coupling. AB - The standard system-plus-reservoir approach used in the study of dissipative systems can be meaningfully generalized to a dissipative coupling involving the momentum, instead of the coordinate: the corresponding equation of motion differs from the Langevin equation, so this is called anomalous dissipation. It occurs for systems where such coupling can indeed be derived from the physical analysis of the degrees of freedom that can be treated as a dissipation bath. Starting from the influence functional corresponding to anomalous dissipation, it is shown how to derive the effective classical potential that gives the quantum thermal averages for the dissipative system in terms of classical-like calculations; the generalization to many degrees of freedom is given. The formalism is applied to a single particle in a double well and to the discrete phi(4) model. At variance with the standard case, the fluctuations of the coordinate are enhanced by anomalous dissipative coupling. PMID- 11736254 TI - Inherent structures in models for fragile and strong glass. AB - An analysis of the dynamics is performed of exactly solvable models for fragile and strong glasses, exploiting the partitioning of the free-energy landscape in inherent structures. The results are compared with the exact solution of the dynamics, by employing the formulation of an effective temperature used in literature. Also, a statistical mechanics formulation is introduced, based upon general statistical considerations, which performs better. Though the considered models are conceptually simple, there is no limit in which the dynamics may be exactly described by a symbolic dynamics of the system moving through consistently weighted inherent structures. PMID- 11736255 TI - Self-consistent quantal treatment of decay rates within the perturbed static path approximation. AB - The framework of the perturbed static path approximation is used to calculate the partition function of a finite Fermi system from a Hamiltonian with a separable two body interaction. Therein, the collective degree of freedom is introduced in self-consistent fashion through a Hubbard-Stratonovich transformation. In this way, all transport coefficients that dominate the decay of a metastable system are defined and calculated microscopically. Otherwise the same formalism is applied as in the Caldeira-Leggett model to deduce the decay rate from the free energy above the so called crossover temperature T(0). PMID- 11736256 TI - Finite driving rates in interface models of Barkhausen noise. AB - We consider a single-interface model for the description of Barkhausen noise in soft ferromagnetic materials. Previously, the model was used only in the adiabatic regime of infinitely slow field ramping. We introduce finite driving rates and analyze the scaling of event sizes and durations for different regimes of the driving rate. Coexistence of intermittency, with nontrivial scaling laws, and finite-velocity interface motion is observed for high enough driving rates. Power spectra show a decay approximately omega(-t), with t<2 for finite driving rates, revealing the influence of the internal structure of avalanches. PMID- 11736257 TI - Cellular automaton model considering the velocity effect of a car on the successive car. AB - In this paper we present a cellular automata model for one-lane traffic flow. The update rules of velocity of a car depend not only on the positions of this car and the car ahead of it, but also on the velocities of the two cars. Using computer simulations we obtain some basic qualitative results and the fundamental diagram of the proposed model. In comparison with those of the existing models in the literature, we find that the fundamental diagram of the proposed model is more consistent with the results measured in the real traffic, and the model is able to reproduce some relevant macroscopic states that are found in the real traffic flow but cannot be predicted by the existing models. PMID- 11736258 TI - Spatial and temporal spectra of noise driven stripe patterns. AB - Spatial and temporal noise power spectra of stripe patterns are investigated, using as a model a Swift-Hohenberg equation with a stochastic term. In particular, the analytical and numerical investigations show: (1) the temporal noise spectra are of 1/f(alpha) form, where alpha=1+(3-D)/4 with D the spatial dimension of the system; (2) that the stochastic fluctuations of the stripe position are subdiffusive. PMID- 11736259 TI - Scaling fields in the two-dimensional Abelian sandpile model. AB - We consider the unoriented two-dimensional Abelian sandpile model from a perspective based on two-dimensional (conformal) field theory. We compute lattice correlation functions for various cluster variables (at and off criticality), from which we infer the field-theoretic description in the scaling limit. We find perfect agreement with the predictions of a c=-2 conformal field theory and its massive perturbation, thereby providing direct evidence for conformal invariance and more generally for a description in terms of a local field theory. The question of the height 2 variable is also addressed, with, however, no definite conclusion yet. PMID- 11736260 TI - Self-affine nature of the stress-strain behavior of thin fiber networks. AB - The stress-strain behavior of toilet paper is studied. We find that the damaged parts of stress-strain curves possess a self-affine scaling invariance. Moreover, we find that the stress-strain behavior and the rupture line roughness are characterized by the same scaling (Hurst) exponent H, which is not universal: rather it changes from sample to sample. The variations on H are mainly due to fluctuations in the paper structure, which are larger than statistical errors within a sample. Furthermore, the same exponent governs the changes in the stress strain curve as the strain rate increases. The fractal damage model is employed to explain experimental observations. PMID- 11736261 TI - Possibility between earthquake and explosion seismogram differentiation by discrete stochastic non-Markov processes and local Hurst exponent analysis. AB - The basic scientific point of this paper is to draw the attention of researchers to new possibilities of differentiation of similar signals having different nature. One example of such kinds of signals is presented by seismograms containing recordings of earthquakes (EQ's) and technogenic explosions (TE's). EQ's are among the most dramatic phenomena in nature. We propose here a discrete stochastic model for possible solution of a problem of strong EQ forecasting and differentiation of TE's from the weak EQ's. Theoretical analysis is performed by two independent methods: by using statistical theory of discrete non-Markov stochastic processes [Phys. Rev. E 62, 6178 (2000)] and the local Hurst exponent. The following Earth states have been considered among them: before (Ib) and during (I) strong EQ, during weak EQ (II) and during TE (III), and in a calm state of Earth's core (IV). The estimation of states I, II, and III has been made on the particular examples of Turkey (1999) EQ's, state IV has been taken as an example of Earth's state before underground TE. Time recordings of seismic signals of the first four dynamic orthogonal collective variables, six various planes of phase portrait of four-dimensional phase space of orthogonal variables and the local Hurst exponent have been calculated for the dynamic analysis of states of systems I-IV. The analysis of statistical properties of seismic time series I-IV has been realized with the help of a set of discrete time-dependent functions (time correlation function and first three memory functions), their power spectra, and the first three points in the statistical spectrum of non Markovity parameters. In all systems studied we have found a bizarre combination of the following spectral characteristics: the fractal frequency spectra adjustable by phenomena of usual and restricted self-organized criticality, spectra of white and color noises and unusual alternation of Markov and non Markov effects of long-range memory, detected earlier [J. Phys. A 27, 5363 (1994)] only for hydrodynamic systems. Our research demonstrates that discrete non-Markov stochastic processes and long-range memory effects play a crucial role in the behavior of seismic systems I-IV. The approaches, permitting us to obtain an algorithm of strong EQ forecasting and to differentiate TE's from weak EQ's, have been developed. PMID- 11736262 TI - Pathological behavior in the spectral statistics of the asymmetric rotor model. AB - The aim of this work is to study the spectral statistics of the asymmetric rotor model (triaxial rigid rotator). The asymmetric top is classically integrable and, according to the Berry-Tabor theory, its spectral statistics should be Poissonian. Surprisingly, our numerical results show that the nearest-neighbor spacing distribution P(s) and the spectral rigidity Delta(3)(L) do not follow Poisson statistics. In particular, P(s) shows a sharp peak at s=1 while Delta(3)(L) for small values of L follows the Poissonian predictions and asymptotically it shows large fluctuations around its mean value. Finally, we analyze the information entropy, which shows a dissolution of quantum numbers by breaking the axial symmetry of the rigid rotator. PMID- 11736263 TI - Noise-enhanced temporal regularity in coupled chaotic oscillators. AB - Existing works on coherence resonance, i.e., the phenomenon of noise-enhanced temporal regularity, focus on excitable dynamical systems such as those described by the FitzHugh-Nagumo equations. We extend the scope of coherence resonance to an important class of nonexcitable dynamical systems: coupled chaotic oscillators. In particular, we argue that, when a system of coupled chaotic oscillators in a noisy environment is viewed as a signal processing unit, the degree of temporal regularity of certain output signals may be modulated by noise and may reach a maximum value at some optimal noise level. Implications to signal processing in biological systems are pointed out. PMID- 11736264 TI - Phase locking in on-off intermittency. AB - Dynamical behavior of on-off intermittency around chaos synchronization desynchronization bifurcation parameter line is investigated in coupled identical chaotic oscillators. Along this parameter line, we find that on-off intermittency can transit from phase-unlocking status to phase-locking one in the phase space of variable differences, which can be regarded as a codimension-two bifurcation, i.e., combinative bifurcations of desynchronization and phase locking. In the phase-locking case, the motions of all oscillators are chaotic and they show on off intermittency with respect to the synchronous manifold, however, spatial phase order of variable differences is clearly established. PMID- 11736265 TI - Influence of noise on scalings for in-out intermittency. AB - We study the effects of noise on a recently discovered form of intermittency, referred to as in-out intermittency. This type of intermittency, which reduces to on-off in systems with a skew product structure, has been found in the dynamics of maps, (ODE) and (PDE) simulations that have symmetries. It shows itself in the form of trajectories that spend a long time near a symmetric state interspersed with short bursts away from symmetry. In contrast to on-off intermittency, there are clearly distinct mechanisms of approach towards and away from the symmetric state, and this needs to be taken into account in order to properly model the long time statistics. We do this by using a diffusion-type equation with a delay integral boundary condition. This model is validated by considering the statistics of a two-dimensional map with and without the addition of noise. PMID- 11736266 TI - Emergence of a dominant unit in a network of chaotic units with a delayed connection change. AB - We study here a model of globally coupled units with adaptive interaction weights that has a delay in the updating rule. Simulations show that the model with such delayed synaptic change exhibits dynamical self organization of network structure. With suitably chosen parameters, "dominant" unit emerges spontaneously, in the sense that the connections from such a unit to almost all of the other units are especially strengthened. Such weight structure facilitates the coherent activity among units. PMID- 11736267 TI - Geometric approach to Lyapunov analysis in Hamiltonian dynamics. AB - As is widely recognized in Lyapunov analysis, linearized Hamilton's equations of motion have two marginal directions for which the Lyapunov exponents vanish. Those directions are the tangent one to a Hamiltonian flow and the gradient one of the Hamiltonian function. To separate out these two directions and to apply Lyapunov analysis effectively in directions for which Lyapunov exponents are not trivial, a geometric method is proposed for natural Hamiltonian systems, in particular. In this geometric method, Hamiltonian flows of a natural Hamiltonian system are regarded as geodesic flows on the cotangent bundle of a Riemannian manifold with a suitable metric. Stability/instability of the geodesic flows is then analyzed by linearized equations of motion that are related to the Jacobi equations on the Riemannian manifold. On some geometric settings on the cotangent bundle, it is shown that along a geodesic flow in question, there exist Lyapunov vectors such that two of them are in the two marginal directions and the others orthogonal to the marginal directions. It is also pointed out that Lyapunov vectors with such properties cannot be obtained in general by the usual method that uses linearized Hamilton's equations of motion. Furthermore, it is observed from numerical calculation for a model system that Lyapunov exponents calculated in both methods, geometric and usual, coincide with each other, independently of the choice of the methods. PMID- 11736268 TI - Scaling properties of bicritical dynamics in unidirectionally coupled period doubling systems in the presence of noise. AB - We study scaling regularities associated with the effects of additive noise on the bicritical behavior of a system of two unidirectionally coupled quadratic maps. A renormalization group analysis of the effects of noise is developed. We outline the qualitative and quantitative differences between the response of the system to random perturbations added to the master subsystem or the slave subsystem. The universal constants determining the rescaling rules for the intensity of the noise sources in the master and slave subsystems are found to be gamma=6.619036... and nu=2.713708..., respectively. A number of computer graphical illustrations for the scaling regularities is presented. We discuss the smearing of the fine structure of the bicritical attractor and the Fourier spectra in the presence of noise, the self-similar structure of the Lyapunov charts on the parameter plane near the bicritical point, and the shift of the threshold of hyperchaos in dependence of the noise intensity. PMID- 11736269 TI - Wada basins and chaotic invariant sets in the Henon-Heiles system. AB - The Henon-Heiles Hamiltonian is investigated in the context of chaotic scattering, in the range of energies where escaping from the scattering region is possible. Special attention is paid to the analysis of the different nature of the orbits, and the the invariant sets, such as the stable and unstable manifolds and the chaotic saddle. Furthermore, a discussion on the average decay time associated to the typical chaotic transients, which are present in this problem, is presented. The main goal of this paper is to show, by using various computational methods, that the corresponding exit basins of this open Hamiltonian are not only fractal, but they also verify the more restrictive property of Wada. We argue that this property is verified by typical open Hamiltonian systems with three or more escapes. PMID- 11736270 TI - Amplitude equations for Rayleigh-Benard convective rolls far from threshold. AB - An extension of the amplitude method is proposed. An iterative algorithm is developed to build an amplitude equation model that is shown to provide precise quantitative results even far from the linear instability threshold. The method is applied to the study of stationary Rayleigh-Benard thermoconvective rolls in the nonlinear regime. In particular, the generation of second and third spatial harmonics is analyzed. Comparison with experimental results and direct numerical calculations is also made and a very good agreement is found. PMID- 11736271 TI - Coupling particles and fields in a diffusive hybrid model. AB - A general scheme to patch together discrete and continuous descriptions of diffusion within the same physical space is studied. In the discrete description, diffusion is described by microscopic random walkers on a lattice; in the continuous description, diffusion is described through the macroscopic diffusion equation. The coupling scheme is based on the mutual exchange of mass flux across the discrete-continuous interface. Detailed tests of the scheme, coupling particle, and field descriptions are particularly illustrative for the diffusion problem. Both the nonequilibrium transport behavior and the equilibrium fluctuations of the combined discrete-continuous system are in agreement with theoretical predictions. PMID- 11736272 TI - Mean-field diffusion-limited aggregation: a "density" model for viscous fingering phenomena. AB - We explore a universal "density" formalism to describe nonequilibrium growth processes, specifically, the immiscible viscous fingering in Hele-Shaw cells (usually referred to as the Saffman-Taylor problem). For that we develop an alternative approach to the viscous fingering phenomena, whose basic concepts have been recently published in a Rapid Communication [Phys. Rev. E 63, 045305(R) (2001)]. This approach uses the diffusion-limited aggregation (DLA) paradigm as a core: we introduce a mean-field DLA generalization in stochastic and deterministic formulations. The stochastic model, a quasicontinuum DLA, simulates Monte Carlo patterns, which demonstrate a striking resemblance to natural Hele Shaw fingers and, for steady-state growth regimes, follow precisely the Saffman Taylor analytical solutions in channel and sector configurations. The relevant deterministic theory, a complete set of differential equations for a time development of density fields, is derived from that stochastic model. As a principal conclusion, we prove an asymptotic equivalency of both the stochastic and deterministic mean-field DLA formulations to the classic Saffman-Taylor hydrodynamics in terms of an interface evolution. PMID- 11736273 TI - Coupled capillary and gravity-driven instability in a liquid film overlying a porous layer. AB - In this work, we study the problem of onset of thermal convection in a fluid layer overlying a porous layer, the whole system being heated from below. We use Brinkman's model to describe the porous medium and determine the corresponding linear stability equations. The eigenvalue problem is solved by means of a modified Galerkin method. The behavior of the critical wave number and temperature gradient is discussed in terms of the various parameters of the system. We also emphasize the influence of the boundary conditions at the upper surface of the fluid layer; in particular, we examine the role of a free surface whose surface tension is temperature dependent (Marangoni effect). Comparison with earlier works is also made. PMID- 11736274 TI - Influence of irradiation on the space-time structure of shock waves. AB - The long-range energy deposition by heavy-ion beams makes new shock wave experiments possible in the laboratory. We have investigated a situation that is of relevance to supernova dynamics in astrophysics, where a shock wave is irradiated by a flux of neutrinos depositing energy throughout the shock wave and surrounding matter, thus changing the behavior of the running shock. We have carried out fluid-dynamical simulations to study generic features of stimulated shock waves. First we consider an idealized case assuming uniform energy deposition into a planar shock wave propagating through an ideal gas. Then we investigate more realistic situations realizable in laboratory experiments with heavy-ion beams. We have found that energy deposition leads to two important effects: acceleration of the shock front and decay of the shock strength. The possibility of laboratory experiments is briefly discussed. PMID- 11736275 TI - Prandtl-number dependence of turbulent flame propagation. AB - Inertial-range cascade phenomenology is used to predict Prandtl-number (Pr) dependencies of turbulent flame properties. A unified picture of turbulent flame structure and burning velocity is developed that encompasses all Pr regimes. Implications of the analysis for gaseous flames (Pr near unity), autocatalytic fronts in liquids (high Pr), and astrophysical flames (low Pr) are noted. PMID- 11736276 TI - Dark solitons in electron-positron plasmas. AB - We demonstrate that a dark soliton is a stable nonlinear coherent mode in electron-positron plasmas. The dark soliton comprises the minimum of the electromagnetic energy density and the minimum of the plasma density. Contrary to a bright soliton, the dark soliton can advect the trapped charged particles. The energy of trapped particles is well above the kinetic energy of the particles in the background plasma. PMID- 11736277 TI - Structure of finite two-dimensional Yukawa lattices: dust crystals. AB - Dust particles in plasmas are often confined near the boundary between the plasma bulk and the sheath where the gravitation is balanced by electrostatic force. To keep dust particles from running away horizontally, an electrostatic potential is usually applied to the electrode surrounding these dusty plasmas and, under appropriate conditions, we have finite two-dimensional lattices of dust particles. Modeling the interaction between dust particles as the isotropic Yukawa interaction, structures of finite two-dimensional Yukawa systems at low temperatures have been analyzed both by numerical simulations and variational methods. The effect of the correlation energy between dust particles is shown to play an important role in the formation of the one-body distribution in these systems. PMID- 11736278 TI - Numerical study of laser wake field generated by two colliding laser beams. AB - The laser wake field generated by two colliding laser beams has been studied numerically. The wake field amplitude is enhanced by a counterpropagating long pulse laser, which has an appropriate frequency difference, and becomes an order of magnitude larger than that of the standard wake field. The field amplitude increases in proportion to the pumping laser intensities until it saturates under the wave breaking limit. The details of the enhanced wake field have been examined at the saturated state. PMID- 11736279 TI - Effects of dust charge fluctuations on current-driven dust-ion-acoustic waves. AB - The effects of fluctuating grain charge on current-driven dust-ion-acoustic modes are investigated in the presence of various physical parameters which are varied in order to describe the behavior of the mode in detail. Variation of the charge clearly influences the ion-acoustic mode by changing of its frequency and growth rate. We report here the existence of a dust charge fluctuation mode in the system, and it is shown that the mode is unstable for a negative charge on the dust per unit volume not exceeding some critical value. PMID- 11736280 TI - Behavior of the H atom velocity distribution function within the shock wave of a hydrogen plasma jet. AB - The evolution of the ground-state hydrogen atom velocity distribution function throughout the stationary shock wave of a supersonic hydrogen plasma jet (3 or =2. This model equation is derived for a situation of two linearly coupled excitations (independently of dimension), and we analytically and numerically study its properties in 2+1 dimensions. We analyze theoretically and demonstrate numerically the dependence of the discrete breather solutions on the amplitude and range of the interactions. We find that complete suppression of the existence thresholds obtained recently for short-range interactions can be achieved beyond a critical value of the amplitude or of the range of the long-range kernel. For supercritical values of the corresponding parameters, staggered branches of solutions are obtained both in theory as well as in the numerical experiment. PMID- 11736296 TI - Addressing and imaging high optical index dielectric ridges in the optical near field. AB - Experimental observation of light coupling between TiO(2) integrated waveguides of subwavelength cross section and pure three-dimensional evanescent light fields is reported. This near-field optical phenomenon is produced by controlling the location of the focusing of a laser beam totally reflected at the surface of the sample. The phenomenon is observed in direct space with a photon scanning tunneling microscope. Dielectric ridges several tens of micrometers long have been efficiently excited with this technique. Upon excitation, the extremities of the linear dielectric wires display intense light spots localized both inside and around the ridge. For ridge lengths up to 30 microm, the observed phenomenon has been reproduced numerically with a parallel implementation based on the three dimensional Green dyadic method. PMID- 11736297 TI - Energy relaxation in nonlinear one-dimensional lattices. AB - We study energy relaxation in thermalized one-dimensional nonlinear arrays of the Fermi-Pasta-Ulam type. The ends of the thermalized systems are placed in contact with a zero-temperature reservoir via damping forces. Harmonic arrays relax by sequential phonon decay into the cold reservoir, the lower-frequency modes relaxing first. The relaxation pathway for purely anharmonic arrays involves the degradation of higher-energy nonlinear modes into lower-energy ones. The lowest energy modes are absorbed by the cold reservoir, but a small amount of energy is persistently left behind in the array in the form of almost stationary low frequency localized modes. Arrays with interactions that contain both a harmonic and an anharmonic contribution exhibit behavior that involves the interplay of phonon modes and breather modes. At long times relaxation is extremely slow due to the spontaneous appearance and persistence of energetic high-frequency stationary breathers. Breather behavior is further ascertained by explicitly injecting a localized excitation into the thermalized arrays and observing the relaxation behavior. PMID- 11736298 TI - Statistics of the Lyapunov exponent in one-dimensional layered systems. AB - Localization of acoustic waves in a one-dimensional water duct containing many randomly distributed air filled blocks is studied. Both the Lyapunov exponent and its variance are computed. Their statistical properties are also explored extensively. The results reveal that in this system a single-parameter scaling is generally inadequate no matter whether the frequency we consider is located in a pass band or in a band gap. This contradicts the earlier observations in an optical case. We compare the results with two optical cases and give a possible explanation of the origin of the different behaviors. PMID- 11736299 TI - Light rays and imaging in wave optics. AB - An interpretation of focusing and image formation on scattering of electromagnetic waves by a dielectric cylinder (a cylindrical lens) is proposed on the basis of the full Maxwell theory. It is centered on analysis of the behavior of integral curves of the Poynting vector here called wave rays. These wave rays cannot intersect so that the focusing and imaging spots are identified with regions of high flow concentration. Two-dimensional examples of wave rays and wave fronts in the scattering of plane and cylindrical electromagnetic waves as well as of Gaussian beams by a dielectric cylinder derived from rigorous solution of the Maxwell equations for incident waves perpendicular to and uniform along the scatterer are given. Their qualitative comparison with geometrical and diffraction approximations are provided. Fixed points and vortex structure of the Poynting flow are investigated. An example of (Gaussian-beam) scattering with transparent multiple internal reflections and multiple wave splitting is given. PMID- 11736300 TI - Ring solitons on vortices. AB - Interaction of a ring dark or antidark soliton (RDS and RADS, respectively) with a vortex is considered in the defocusing nonlinear Schrodinger equation with cubic (for RDS) or saturable (for RADS) nonlinearities. By means of direct simulations, it is found that the interaction gives rise to either an almost isotropic or a spiral-like pattern. A transition between them occurs at a critical value of the RDS or RADS amplitude, the spiral pattern appearing if the amplitude exceeds the critical value. An initial ring soliton created on top of the vortex splits into a pair of rings moving inward and outward. In the subcritical case, the inbound ring reverses its polarity, bouncing from the vortex core, without conspicuous effect on the core. In the transcritical case, the bounced ring soliton suffers a spiral deformation, while the vortex changes its position and structure and also loses its axial symmetry. Through a variational-type approach to the system's Hamiltonian, we additionally find that the vortex-RDS and vortex-RADS interactions are, respectively, attractive and repulsive. Simulations with the vortex placed eccentrically with respect to the RDS or RADS reveal the generation of strongly localized multispot dark and/or antidark coherent structures. The occurrence of spiral-like patterns in many numerical experiments prompted an attempt to generate a spiral dark soliton, but the latter is found to suffer a core instability that converts it into a rotating dipole emitting waves in the outward direction. PMID- 11736301 TI - Fifth-order corrected field descriptions of the Hermite-Gaussian (0,0) and (0,1) mode laser beam. AB - In this paper, we extend the work of Barton and Alexander [J. App. Phys. 66, 2800 (1989)] on the fifth-order corrected field expressions for a Hermite-Gaussian (0,0) mode laser beam to more general cases with adjustable parameters. The parametric dependence of the electron dynamics is investigated by numerical methods. Finally, the fifth-order corrected field equations for the Hermite Gaussian (0,1) mode are also presented. PMID- 11736302 TI - Breaking Abbe's diffraction resolution limit in fluorescence microscopy with stimulated emission depletion beams of various shapes. AB - We report on the generation of various hole-centered beams in the focal region of a lens and investigate their effectiveness to break the diffraction barrier in fluorescence microscopy by stimulated emission. Patterning of the phase of the stimulating beam across the entrance pupil of the objective lens produces point spread-functions with twofold, fourfold, and circular symmetry, which narrow down the focal spot to 65-100 nm. Comparison with high-resolution confocal images exhibits a resolution much beyond the diffraction barrier. Particles that are only 65-nm apart are resolved with focused light. PMID- 11736303 TI - Collapse arresting in an inhomogeneous two-dimensional nonlinear Schrodinger model. AB - Collapse of (2+1)-dimensional beams in the inhomogeneous two-dimensional cubic nonlinear Schrodinger equation is analyzed numerically and analytically. It is shown that in the vicinity of a narrow attractive inhomogeneity, the collapse of beams that in a homogeneous medium would collapse may be arrested under certain circumstances. PMID- 11736304 TI - Effects of transport memory and nonlinear damping in a generalized Fisher's equation. AB - Memory effects in transport require, for their incorporation into reaction diffusion investigations, a generalization of traditional equations. The well known Fisher's equation, which combines diffusion with a logistic nonlinearity, is generalized to include memory effects, and traveling wave solutions of the equation are found. Comparison is made with alternative generalization procedures. PMID- 11736305 TI - Third-order implementation and convergence of the strong-property-fluctuation theory in electromagnetic homogenization. AB - The strong-property-fluctuation theory (SPFT) has been widely applied under the second-order approximation (also known as the bilocal approximation) to estimate the constitutive properties of effectively homogeneous composite mediums. A third order mass operator approximation is developed here. The convergence of the long wavelength, bilocally-approximated SPFT is demonstrated for isotropic chiral composite mediums, as well as for chiroferrite composite mediums. PMID- 11736306 TI - Solitary waves in systems with separated Bragg grating and nonlinearity. AB - The existence and stability of solitons in a dual-core optical waveguide, in which one core has Kerr nonlinearity while the other one is linear with a Bragg grating written on it, are investigated. The system's spectrum for the frequency omega of linear waves always contains a gap. If the group velocity c in the linear core is zero, it also has two other, upper and lower (in terms of omega) gaps. If c not equal to 0, the upper and lower gaps do not exist in the rigorous sense, as each overlaps with one branch of the continuous spectrum. When c=0, a family of zero-velocity soliton solutions, filling all the three gaps, is found analytically. Their stability is tested numerically, leading to a conclusion that only solitons in an upper section of the upper gap are stable. For c not equal to 0, soliton solutions are sought for numerically. Stationary solutions are only found in the upper gap, in the form of unusual solitons, which exist as a continuous family in the former upper gap, despite its overlapping with one branch of the continuous spectrum. A region in the parameter plane (c,omega) is identified where these solitons are stable; it is again an upper section of the upper gap. Stable moving solitons are found too. A feasible explanation for the (virtual) existence of these solitons, based on an analytical estimate of their radiative-decay rate (if the decay takes place), is presented. PMID- 11736307 TI - Accessing the dynamics of large many-particle systems using the stochastic series expansion. AB - The stochastic series expansion (SSE) method is a quantum Monte Carlo (QMC) technique working directly in the imaginary time continuum and thus avoiding "Trotter discretization" errors. Using a nonlocal "operator-loop update," it allows one to treat large quantum mechanical systems of many thousand sites. In this paper we first give a comprehensive review on SSE and present benchmark calculations of SSE scaling behavior with system size and inverse temperature, and compare it to the loop algorithm, whose scaling is known to be one of the best of all QMC methods. Finally we introduce an efficient algorithm to measure Green's functions and thus dynamical properties within SSE. PMID- 11736308 TI - Pore-scale modeling of saturated permeabilities in random sphere packings. AB - We use two pore-scale approaches, lattice-Boltzmann (LB) and pore-network modeling, to simulate single-phase flow in simulated sphere packings that vary in porosity and sphere-size distribution. For both modeling approaches, we determine the size of the representative elementary volume with respect to the permeability. Permeabilities obtained by LB modeling agree well with Rumpf and Gupte's experiments in sphere packings for small Reynolds numbers. The LB simulations agree well with the empirical Ergun equation for intermediate but not for small Reynolds numbers. We suggest a modified form of Ergun's equation to describe both low and intermediate Reynolds number flows. The pore-network simulations agree well with predictions from the effective-medium approximation but underestimate the permeability due to the simplified representation of the porous media. Based on LB simulations in packings with log-normal sphere-size distributions, we suggest a permeability relation with respect to the porosity, as well as the mean and standard deviation of the sphere diameter. PMID- 11736309 TI - Fully compact higher-order computation of steady-state natural convection in a square cavity. AB - The flow in a thermally driven square cavity with adiabatic top and bottom walls and differentially heated vertical walls for a wide range of Rayleigh numbers (10(3)< or =Ra< or =10(7)) has been computed with a fourth-order accurate higher order compact scheme, which was used earlier only for the stream-function vorticity (psi-omega) form of the two-dimensional steady-state Navier-Stokes equations. The boundary conditions used are also compact and of identical accuracy. In particular, a compact fourth-order accurate Neumann boundary condition has been developed for temperature at the adiabatic walls. The treatment of the derivative source term is also compact and has been done in such a way as to give fourth-order accuracy and easy assimilation with the solution procedure. As the discretization for the psi-omega formulation, boundary conditions, and source term treatment are all fourth-order accurate, highly accurate solutions are obtained on relatively coarser grids. Unlike other compact solution procedure in literature for this physical configuration, the present method is fully compact and fully higher-order accurate. Also, use of conjugate gradient and hybrid biconjugate gradient stabilized algorithms to solve the symmetric and nonsymmetric algebraic systems at every outer iteration, ensures good convergence behavior of the method even at higher Rayleigh numbers. PMID- 11736310 TI - Fourier acceleration of Langevin molecular dynamics. AB - Fourier acceleration has been successfully applied to the simulation of lattice field theories for more than a decade. In this paper, we extend the method to the dynamics of discrete particles moving in a continuum. Although our method is based on a mapping of the particles' dynamics to a regular grid so that discrete Fourier transforms may be taken, it should be emphasized that the introduction of the grid is a purely algorithmic device and that no smoothing, coarse-graining, or mean-field approximations are made. The method thus can be applied to the equations of motion of molecular dynamics (MD) or its Langevin or Brownian variants. For example, in Langevin MD simulations our acceleration technique permits a straightforward spectral decomposition of forces so that the long wavelength modes are integrated with a longer time step, thereby reducing the time required to reach equilibrium or to decorrelate the system in equilibrium. Speedup factors of up to 30 are observed relative to pure (unaccelerated) Langevin MD. As with acceleration of critical lattice models, even further gains relative to the unaccelerated method are expected for larger systems. Preliminary results for Fourier-accelerated molecular dynamics are presented in order to illustrate the basic concepts. Possible extensions of the method and further lines of research are discussed. PMID- 11736311 TI - Unconditionally stable algorithms to solve the time-dependent Maxwell equations. AB - Based on the Suzuki product-formula approach, we construct a family of unconditionally stable algorithms to solve the time-dependent Maxwell equations. We describe a practical implementation of these algorithms for one-, two-, and three-dimensional systems with spatially varying permittivity and permeability. The salient features of the algorithms are illustrated by computing selected eigenmodes and the full density of states of one-, two-, and three-dimensional models and by simulating the propagation of light in slabs of photonic band-gap materials. PMID- 11736312 TI - Renormalization group analysis of autoregressive processes and fractional noise. AB - A renormalization group analysis is applied to autoregressive processes with an infinite series of coefficients. A simple fixed point is given by a random walk, and a second class is found that is proportional to the high order coefficients of fractional autoregressive integrated moving average (ARIMA) processes. The approach might be useful to detect nonstationarity in autoregressive processes. PMID- 11736313 TI - Equivalence of the Ashkin-Teller and the four-state Potts-glass models of neural networks. AB - We show that for a particular choice of the coupling parameters the Ashkin-Teller spin-glass neural network model with the Hebb learning rule and one condensed pattern yields the same thermodynamic properties as the four-state anisotropic Potts-glass neural network model. This equivalence is not seen at the level of the Hamiltonians. PMID- 11736314 TI - Q learning in the minority game. AB - We present a numerical investigation of the minority game model, where the dynamics of the agents is described by the Q-learning algorithm. The numerical results show that the Q-learning dynamics is suppressing the "crowd effect," which is characteristic of the minority game with standard inductive dynamics, and it converges to a stationary state that is close to the optimal Nash equilibrium of the game. PMID- 11736315 TI - Correlation properties of binary spatiotemporal chaotic sequences and their application to multiple access communication. AB - This paper studies the correlation property and the spectral density of the binary spatiotemporal chaotic sequences that are generated by the one-way coupled map lattice. The results show that this kind of chaotic sequences possesses excellent randomlike property and could be used directly in the spread spectrum multiple access (SSMA) communications. The multiple access interference in the additional white Gaussian noise background is then analyzed, and the corresponding formulas are presented. The simulation and computation results indicate that the communication system adopting such spreading sequences possesses as good performance as the one employing Gold sequences. But the former has larger capacity and higher complexity. Therefore, the binary spatiotemporal chaotic sequences are good candidates for the spreading sequences in SSMA communications. PMID- 11736316 TI - Unimodal order in the image of the simplest equivariant chaotic system. AB - The simplest equivariant chaotic dynamics is investigated in terms of its image, i.e., under the 2-->1 mapping allowing one to obtain a projection of the dynamics without any residual symmetry. The inversion symmetry is therefore deleted. The bifurcation diagram can thus be predicted from the unimodal order although the first-return map computed in the original phase space exhibits three critical points. This feature is the same as the one observed on the Burke and Shaw system although this latter system has a rotation symmetry. PMID- 11736317 TI - Energy spectrum of grid-generated He II turbulence. AB - A grid of bars towed through a sample of He II produces both superfluid turbulence and classical hydrodynamic turbulence. The two velocity fields-in the normal fluid and in the superfluid-have been observed to have the same energy spectral density over a large range of scales. Here, we introduce a characteristic scale l(q)=2 pi(epsilon/kappa(3))(-1/4), where epsilon is the rate of turbulent energy dissipation per unit volume, and note that the energy spectrum in superfluid turbulence depends also on the quantum of circulation kappa, for wave numbers k>k(q) identical with 2 pi/l(q). We propose that the spectral density in this range is of the form straight phi(k)=C epsilon kappa( 1)k(-3), where C is the three-dimensional Kolmogorov constant in classical turbulence. This form is consistent with recent experiments in the temperature range 1.2 K